Stem Cell Research & Therapy•2026•DOI: 10.1186/s13287-026-05051-z
Background Intervertebral disc degeneration (IVDD) is a major cause of low back pain, characterized by mitochondrial dysfunction and impaired mitophagy in nucleus pulposus-derived mesenchymal stem cells (NPMSCs). Sirtuin 1 (SIRT1), a key regulator of mitochondrial quality control, is downregulated in degenerated discs. Salidroside (Sal), a natural compound from Rhodiola rosea, has shown potential in enhancing mitophagy, but its mechanism in IVDD remains unclear. Methods Using network pharmacology, molecular docking, and dynamics simulations, we identified SIRT1 as a key target of Sal. Human and rat NPMSCs were isolated and treated with tert-butyl hydroperoxide (TBHP) to induce degeneration. In vitro assays included CCK-8, EdU, SA-β-Gal, JC-1, Western blot, immunofluorescence, and transmission electron microscope (TEM). An in vivo rat IVDD model was established via needle puncture and treated with Sal and/or the autophagy inhibitor 3-MA. Histological, immunohistochemical, and imaging analyses were performed to evaluate IVDD. Results Sal bound stably to SIRT1 and activated SIRT1/FOXO3 pathway, promoting mitophagic flux, reducing reactive oxygen species accumulation, and suppressing apoptosis in NPMSCs. SIRT1 knockdown or 3-MA treatment abolished Sal’s protective effects. In vivo, Sal treatment preserved disc height, reduced apoptosis, and enhanced mitophagy, while 3-MA exacerbated degeneration.
Stem Cell Research & Therapy•2026•DOI: 10.1186/s13287-026-05073-7
Exosomes have emerged as critical mediators of intercellular and inter-organ communication in bone biology. Secreted by bone-resident cells such as osteoblasts, osteoclasts, osteocytes, and mesenchymal stem cells (MSCs), these nanosized vesicles carry diverse molecular cargos that regulate bone remodeling, regeneration, and skeletal homeostasis. In addition to mediating local communication within the bone microenvironment, exosomes also participate in systemic crosstalk communication between bone and other tissues, including skeletal muscle, adipose tissue, gut microbiota, the immune system, the nervous system, and vasculature. Disruption of these exosome-mediated pathways contributes to the development and progression of bone diseases, including osteoporosis, osteoarthritis, osteonecrosis of the femoral head, and bone metastases. This review summarizes current advances in exosome-mediated signaling in both physiological and pathological contexts, with particular emphasis on their roles as biomarkers, therapeutic agents, and drug delivery vehicles. We also discuss the emerging contribution of artificial intelligence (AI) to exosome research, especially in biomarker discovery, disease classification, and target identification, as well as the major challenges that currently limit clinical translation. Together, these insights highlight the potential of exosome-based strategies for precision medicine in bone diseases.
Stem Cell Research & Therapy•2025•DOI: 10.1186/s13287-025-04396-1
Background One major factor contributing to infertility in women of childbearing age is premature ovarian insufficiency (POI). Exosomes produced from human umbilical cord mesenchymal stem cells (hUMSC-Exos) have drawn a lot of attention lately as a potential treatment for ovarian dysfunction brought on by POI. However, its therapeutic mechanism is still unclear and needs further exploration.
Methods POI model was established by intraperitoneal injection of cyclophosphamide (CTX) in female Wistar rats. These POI rats were treated with hUMSC-Exos for one week. In addition to in vivo experiments, in vitro POI models were also established. In vitro experiments, theca interstitial cells (TICs) treated with CTX were exposed to normal as well as let-7a-5p inhibitory hUMSC-Exos. The ovary structure, morphology, endocrine function, and reproductive ability of POI rats were observed by H&E staining and ELISA. Western blot, immunofluorescence staining (IF), and quantitative real-time polymerase chain reaction (qRT-PCR) were used to evaluate the autophagy-related indexes in ovary and TICs of POI rats in each group.
Results CTX induced abnormalities of ovarian morphology, structure, endocrine, and reproductive function in rats, and accompanied by autophagy of TICs. Notably, hUMSC-Exos diminishes ovarian structural and functional damage in POI rats and TICs autophagy via targeting the AMPK/mTOR pathway. Furthermore, downregulating let-7a-5p in hUMSC-Exos weakened their ability to prevent TICs autophagy.
Conclusions Overall, the findings suggested that hUMSC-Exos improves ovarian function in POI rats by inhibiting TICs autophagy via the let-7a-5p/AMPK/mTOR pathway. Our study provided further evidence that POI patients can benefit from hUMSC-Exos-mediated therapy.
Stem Cell Research & Therapy•2025•DOI: 10.1186/s13287-025-04806-4
Background The human umbilical cord (hUC)–mesenchymal stem cell (MSC) secretome (SCT) is a cell-free therapy that may emerge as a novel therapeutic strategy for hair loss prevention. Here, we aimed to elucidate the underlying mechanisms through which SCT regulates hair growth and cycle transition. Methods Using C57BL/6 mice, ex vivo follicles, and cell experiments, we studied the effects and mechanisms of SCT on hair growth and cycling using untargeted metabolomics and phosphoproteomics. A three-month double-blind clinical study was conducted to validate the effects of SCT on human hair. Results SCT promotes the telogen-to-anagen transition, hair thickening, and elongation of the vibrissae in mice; regulates dermal papilla cells and hair matrix cells through cysteine and methionine metabolism; and stimulates methylthioadenosine synthesis in hair matrix cells by activating the PI3K/AKT/mTOR signaling pathway. Clinical studies demonstrated that SCT increased human hair density and average hair diameter. Scalp physiological tests and subjective feedback indicated no related adverse reactions on the scalp or hair. Conclusions SCT promoted hair growth, thickening, and the hair follicle cycle via the PI3K/AKT/mTOR signaling pathway. This research provides a basis for the application of cell-free alternatives in hair care and hair loss prevention.
Stem Cell Research & Therapy•2025•DOI: 10.1186/s13287-025-04366-7
Background Osteoarthritis (OA) is a common degenerative disease caused by multiple pathological mechanisms wherein subchondral bone malfunction plays a substantial role. Recently, subchondral (SC) injection of orthobiologics has been attracting growing interest albeit the mainstream delivery method of mesenchymal stem cells (MSCs) is through intra-articular (IA). This study investigates the effect of SC injection of human umbilical cord mesenchymal stem cells (UCMSCs) on OA and its possible therapeutic mechanism compared to IA injection. Methods Male Sprague-Dawley rats with anterior cruciate ligament transection (ACLT) received saline or UCMSC injections via SC or IA. Consecutive injections once a week for three weeks and withdrawal for another four weeks, followed by Radiographical scanning, histopathological, immunohistochemical, and terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick-end labelling (TUNEL) staining. Cell counting Kit-8 (CCK-8) assay, alkaline phosphatase (ALP), alizarin red staining (ARS), TUNEL, flow cytometry, quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting were employed in TNFα-induced MC3T3-E1 cells to illustrate the exact pathogenesis mechanism. Results IA and SC UCMSC injections preserved cartilage, synovium, and subchondral bone parameters like trabecular bone volume fraction (BV/TV). SC injection uniquely improved Trabecular separation (Tb.Sp) and Trabecular number (Tb.N). SC and IA injections of UCMSCs demonstrated equivalent efficacy in promoting osteoblastic bone formation and attenuating aberrant angiogenesis of subchondral bone. In addition, we demonstrated that osteoblast apoptosis and Smad2-dependent TGF-beta (TGF-β) are crucial and interactive subchondral bone pathological features in OA. In vivo and vitro studies further revealed that UCMSCs inhibited excessive TGF-β/pSmad2 signaling to regulate osteoblast apoptosis and bone remodeling, thereby ameliorating OA progression. These findings suggest that SC injection of UCMSCs is a promising therapeutic strategy for OA, potentially offering advantages over IA injection in improving subchondral bone microarchitecture.
Stem Cell Research & Therapy•2025•DOI: 10.1186/s13287-025-04560-7
The rising prevalence of alopecia poses a significant challenge for both clinicians and researchers. As the global incidence of hair loss continues to increase, research into hair biology and regenerative mechanisms has gained considerable attention. However, current treatment options for alopecia are often constrained by limited efficacy and notable adverse effects. This underscores an urgent need for innovative therapeutic strategies to address these gaps. Adipose-derived stem cells (ADSCs), a subset of mesenchymal stem cells, represent a promising new approach in the treatment of alopecia. This review provides a detailed examination of the fundamental properties of ADSCs and their derivatives, exploring their mechanisms of action in alopecia therapy. Analysis of the efficacy of ADSCs and their derivatives in both preclinical and clinical settings highlight their potential to stimulate hair regeneration. Additionally, the review discusses various pre-treatment methods designed to enhance the regenerative capacity of ADSCs in hair growth, elucidating the mechanisms involved. The review also addresses the challenges and future directions for the use of ADSCs in alopecia treatment, aiming to offer valuable insights for both theoretical research and clinical practice. Ultimately, this work seeks to contribute to the development of more effective treatment regimens for alopecia.
Stem Cell Research & Therapy•2025•DOI: 10.1186/s13287-025-04326-1
Suppressing bone mesenchymal stem cell (BMSC) ferroptosis is expected to optimize BMSCs-based therapy for intervertebral disc degeneration (IVDD). Our previous study revealed that Prominin-2 could protect against ferroptosis by decreasing cellular Fe2+ content and inhibiting transcription regulator protein BACH1 (BACH1) expression. In this study we probed the molecular mechanisms underlying the Prominin-2/BACH1 pathway in BMSC ferroptosis. Using an array of in vitro and in vivo experiments we found that heat shock factor protein 1 (HSF1) activates PROM2 (encoding protein Prominin-2) transcription and elevated Prominin-2 expression. Furthermore, we showed that Prominin-2 attenuates ferroptosis induced by tert-butyl hydroperoxide (TBHP) through promoting BACH1 ubiquitination and degradation. Inhibition of BACH1 expression reversed TBHP-stimulated down expression of glutaminase kidney isoform, mitochondrial (GLS), which plays a crucial role in protecting BMSCs against ferroptosis. Targeting the Prominin-2/BACH1 axis has also been shown to improve BMSC survival post-transplantation and mitigate IVDD progression by inhibiting ferroptosis. Our results support a new mechanistic insight into the regulation of the Prominin-2/BACH1/GLS pathway in BMSC ferroptosis. These finding could lead to potential therapeutic targets to improve the survival of engrafted BMSCs under oxidative stress circumstances.
Stem Cell Research & Therapy•2025•DOI: 10.1186/s13287-025-04267-9
Background: In premature newborn infants, preterm white matter injury (PWMI) causes motor and cognitive disabilities. Accumulating evidence suggests that PWMI may result from defected differentiation of oligodendrocyte precursor cells (OPCs) and impaired maturation of oligodendrocytes. However, the underlying mechanisms remain unclear. Methods: Using RNAscope, we analyzed the expression level of RNA-binding protein LIN28A in individual OPCs. Knockout of one or both alleles of Lin28a in OPCs was achieved by administrating tamoxifen to NG2CreER::Ai14::Lin28aflox/+ or NG2CreER::Ai14::Lin28aflox/flox mice. Lentivirus expressing FLEX-Lin28a was used in NG2CreER mice to overexpress LIN28A in OPCs. A series of behavioral tests were performed to assess the cognitive functions of mice. Two-tailed unpaired t-tests was carried out for statistical analysis between groups. Results: We found that the expression of Lin28a was decreased in OPCs in a PWMI mouse model. Knockout of one or both alleles of Lin28a in OPCs postnatally resulted in reduced OPC differentiation, decreased myelinogenesis and impaired cognitive functions. Supplementing LIN28A in OPCs postnatally was able to promote OPC differentiation and enhance myelinogenesis, thus rescuing the cognitive functions in PWMI mice. Conclusion: Our study reveals that LIN28A is critical in regulating postnatal myelinogenesis. Overexpression of LIN28A in OPCs rescues cognitive deficits in PWMI mice by promoting myelinogenesis, thus providing a potential strategy for the treatment of PWMI.
Stem Cell Research & Therapy•2024•DOI: 10.1186/s13287-024-03761-w
Mesenchymal stem cells (MSCs) are multipotent cells that can differentiate into cells of different lineages to form mesenchymal tissues, which are promising in regard to treatment for bone diseases. Their osteogenic differentiation is under the tight regulation of intrinsic and extrinsic factors. Transforming growth factor β (TGF-β) is an essential growth factor in bone metabolism, which regulates the differentiation of MSCs. However, published studies differ in their views on whether TGF-β signaling regulates the osteogenic differentiation of MSCs positively or negatively. The controversial results have not been summarized systematically and the related explanations are required. Therefore, we reviewed the basics of TGF-β signaling and summarized how each of three isoforms regulates osteogenic differentiation. Three isoforms of TGF-β (TGF-β1/β2/β3) play distinct roles in regulating osteogenic differentiation of MSCs. Additionally, other possible sources of conflicts are summarized here. Further understanding of TGF-β signaling regulation in MSCs may lead to new applications to promote bone regeneration and improve therapies for bone diseases.
Stem Cell Research & Therapy•2024•DOI: 10.1186/s13287-024-03725-0
Background Wharton’s jelly-derived mesenchymal stem cells (WJ-MSCs) hold great therapeutic potential in regenerative medicine. Therefore, it is crucial to establish a Good Manufacturing Practice (GMP)-compliant methodology for the isolation and culture of WJ-MSCs. Through comprehensive research, encompassing laboratory-scale experiments to pilot-scale studies, we aimed to develop standardized protocols ensuring the high yield and quality of WJ-MSCs manufacturing. Methods Firstly, optimization of parameters for the enzymatic digestion method used to isolate WJ-MSCs was conducted. These parameters included enzyme concentrations, digestion times, seeding densities, and culture media. Additionally, a comparative analysis between the explant method and the enzymatic digestion method was performed. Subsequently, the consecutive passaging of WJ-MSCs, specifically up to passage 9, was evaluated using the optimized method. Finally, manufacturing processes were developed and scaled up, starting from laboratory-scale flask-based production and progressing to pilot-scale cell factory-based production. Furthermore, a stability study was carried out to assess the storage and use of drug products (DPs). Results The optimal parameters for the enzymatic digestion method were a concentration of 0.4 PZ U/mL Collagenase NB6 and a digestion time of 3 h, resulting in a higher yield of P0 WJ-MSCs. In addition, a positive correlation between the weight of umbilical cord tissue and the quantities of P0 WJ-MSCs has been observed. Evaluation of different concentrations of human platelet lysate revealed that 2% and 5% concentrations resulted in similar levels of cell expansion. Comparative analysis revealed that the enzymatic digestion method exhibited faster outgrowth of WJ-MSCs compared to the explant method during the initial passage. Passages 2 to 5 exhibited higher viability and proliferation ability throughout consecutive passaging. Moreover, scalable manufacturing processes from the laboratory scale to the pilot scale were successfully developed, ensuring the production of high-quality WJ-MSCs. Multiple freeze-thaw cycles of the DPs led to reduced cell viability and viable cell concentration. Subsequent thawing and dilution of the DPs resulted in a significant decrease in both metrics, especially when stored at 20–27 °C.
Stem Cell Research & Therapy•2024•DOI: 10.1186/s13287-024-03943-6
Although liver transplantation (LT) is an effective strategy for end-stage liver diseases, the shortage of donor organs and the immune rejection hinder its widespread implementation in clinical practice. Mesenchymal stem cells (MSCs) transplantation offers a promising approach for patients undergoing liver transplantation due to their immune regulatory capabilities, hepatic protection properties, and multidirectional differentiation potential. In this review, we summarize the potential applications of MSCs transplantation in various LT scenarios. MSCs transplantation has demonstrated effectiveness in alleviating hepatic ischemia-reperfusion injury, enhancing the viability of liver grafts, preventing acute graft-versus-host disease, and promoting liver regeneration in split LT therapy. We also discuss the clinical progress, and explore the immunomodulatory functions of MSCs in response to both adaptive and innate immune responses. Furthermore, we emphasize the interactions between MSCs and different immune cells, including T cells, B cells, plasma cells, natural killer cells, dendritic cells, Kupffer cells, and neutrophils, to provide new insights into the immunomodulatory properties of MSCs in adoptive cell therapy.
Stem Cell Research & Therapy•2024•DOI: 10.1186/s13287-024-03855-5
Background Hematopoietic stem and progenitor cells (HSPCs) mobilize from bone marrow to peripheral blood in response to stress. The impact of alloresponse-induced stress on HSPCs mobilization in human liver transplantation (LTx) recipients remains under-investigated.
Methods Peripheral blood mononuclear cell (PBMC) samples were longitudinally collected from pre- to post-LTx for one year from 36 recipients with acute rejection (AR), 74 recipients without rejection (NR), and 5 recipients with graft-versus-host disease (GVHD). 28 PBMC samples from age-matched healthy donors were collected as healthy control (HC). Multi-color flow cytometry (MCFC) was used to immunophenotype HSPCs and their subpopulations. Donor recipient-distinguishable major histocompatibility complex (MHC) antibodies determined cell origin.
Results Before LTx, patients who developed AR after transplant contained more HSPCs in PBMC samples than HC, while the NR group patients contained fewer HSPCs than HC. After LTx, the HSPC ratio in the AR group sharply decreased and became less than HC within six months, and dropped to a comparable NR level afterward. During the one-year follow-up period, myeloid progenitors (MPs) biased differentiation was observed in all LTx recipients who were under tacrolimus-based immunosuppressive treatment. During both AR and GVHD episodes, the recipient-derived and donor-derived HSPCs mobilized into the recipient’s blood-circulation and migrated to the target tissue, respectively. The HSPCs percentage in blood reduced after the disease was cured.
Conclusions A preoperative high HSPC ratio in blood characterizes recipients who developed AR after LTx. Recipients exhibited a decline in blood-circulating HSPCs after transplant, the cells mobilized into the blood and migrated to target tissue during alloresponse.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2026010
Persicaria chinensis, a well-known traditional Chinese medicinal herb that is both edible and medicinal, has been widely acknowledged for its therapeutic effects, such as anti-inflammatory, antioxidant, and antitumor activities. However, the role of miRNAs from this plant in the cross-kingdom regulation of human diseases has not been investigated. In this study, we analyze the miRNA expression profile of P. chinensis using high-throughput sequencing and identify a total of 673 miRNAs, including 422 novel miRNAs that are unique to this plant and 251 conserved miRNAs. Among the conserved miRNAs, pch-miR319a is found to be the most abundant. Moreover, food-oriented pch-miR319a accumulates in the uterus and tumors and exhibits a rich repertoire of target genes within cancer-related pathways, demonstrating significant cross-kingdom regulatory potential. Utilizing the dual-luciferase reporter gene assay, we demonstrate that pch-miR319a from P. chinensis targets the Itga3 gene, which is associated with cervical cancer progression. Overexpression of pch-miR319a significantly decreases the viability, migration, and induces apoptosis of HeLa cervical cancer cells in vitro. Moreover, in a syngeneic mouse tumor model of cervical cancer, treatment with pch-miR319a effectively inhibits tumor growth and downregulates the expressions of ITGA3 and the proliferation marker Ki-67. Our study highlights the potential of pch-miR319a from P. chinensis as a novel therapeutic agent for cervical cancer by targeting ITGA3 and provides new insights into the cross-kingdom regulatory mechanisms of plant miRNAs in human diseases.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2026037
Breast cancer (BC) is the most common malignancy among women, with approximately 2.3 million new cases diagnosed annually, accounting for approximately 11.6% of all cancer cases worldwide. Distant metastasis is the primary cause of mortality in BC patients, with nearly 50% of patients ultimately developing metastatic disease. The predominant metastatic sites of BC include the lung, liver, brain, and bone, each exhibiting distinct biological characteristics that drive the organ-specific tropism of cancer cells. Among these, brain metastasis represents a significant cause of mortality in BC patients and is particularly prevalent in those with human epidermal growth factor receptor 2 (HER2)-positive or triple-negative breast cancer (TNBC) subtypes. Breast cancer brain metastasis (BCBM) can manifest in three forms: choroid plexus metastasis (rare), leptomeningeal metastasis (approximately 8%), and parenchymal metastasis, the most common presentation, with multiple lesions in 78% of cases and solitary lesions in 14%. Distinct anatomical regions of the brain provide different micro-environments, which in turn shape epidemiological patterns, biological behaviors, and therapeutic vulnerabilities of metastatic cancer. With the continuous advancement of systemic therapies and imaging surveillance, brain metastases from BC have become increasingly prevalent, accounting for approximately 10%–30% of all metastatic breast cancer (MBC) cases. The continuous progression of BCBM often compromises patients’ cognitive and sensory functions, leading to neurological impairment and severely limiting quality of life (QOL). Notably, the mortality rate within one year after diagnosis remains at 80%. Current therapeutic strategies for BCBM primarily include surgery, whole-brain radiotherapy (WBRT), stereotactic radiosurgery (SRS), chemotherapy, or combinations thereof. Although these approaches provide some clinical benefit, the efficacy remains limited due to the blood-brain barrier (BBB), which restricts drug penetration and contributes to chemoresistance. Therefore, elucidating the molecular mechanisms underlying BCBM is imperative to identify novel diagnostic biomarkers and therapeutic targets, with the ultimate goal of improving treatment efficacy and patient prognosis. Bioinformatics provides a powerful platform and data foundation for exploring the mechanisms of tumor initiation and progression. High-throughput platforms for gene expression analysis have gained significant popularity, with next-generation sequencing (NGS) and microarray analysis now widely applied as essential tools in medical oncology. These techniques have diverse clinical applications, including molecular cancer classification, prediction of therapeutic response, prognostic assessment, molecular diagnostics, and the discovery of novel drugs and therapeutic targets. Weighted gene coexpression network analysis (WGCNA) has been widely applied in studies of gene regulatory networks, biomarker discovery, and elucidation of the molecular mechanisms underlying complex phenotypes. In this study, we utilized the BCBM microarray dataset GSE43837. We performed differential expression analysis and WGCNA clustering using the R packages limma and WGCNA to identify potential gene modules and candidate targets. GSE43837 consists of 19 nonmetastatic primary breast tumor samples and 19 breast cancer brain metastasis samples. Differential expression analysis, with thresholds set at |logFC| > 1 and P < 0.05, identified 245 upregulated and 188 downregulated genes (Supplementary Table S1 and Supplementary Figure S1A). WGCNA further confirmed that the constructed network satisfied the scale-free topology criterion, with the optimal soft-threshold power determined to be 14 based on model fit and mean connectivity (Supplementary Figure S1B). Using the dynamic tree cut method, we clustered genes into multiple modules, each representing a group of coexpressed genes with varying degrees of correlation among modules (Supplementary Figure S1C,D). Notably, the midnightblue and black modules showed stronger correlations, and a significant positive relationship was observed between gene significance (GS) and module membership (MM) within these modules (Supplementary Figure S1E). This finding suggests that the core genes in these modules are highly representative and stable within the coexpression network. A total of 89 BCBM-related candidate genes were extracted from these key modules (Supplementary Table S2). To further identify key feature genes associated with BCBM, we applied two machine learning methods, LASSO regression and random forest (RF), to the 29 overlapping genes obtained from the intersection of DEGs and hub module genes (Figure 1A and Supplementary Table S3). In the LASSO regression analysis, the optimal penalty parameter λ was determined by cross-validation, yielding a set of candidate genes with nonzero regression coefficients (Figure 1B). Concurrently, in the RF model, 500 decision trees were constructed, and the classification ...
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2025253
Kidney renal clear cell carcinoma (KIRC) is the most common and aggressive subtype of renal cell carcinoma and is characterized by poor prognosis and high molecular heterogeneity. Long noncoding RNAs (lncRNAs) have emerged as crucial regulators in cancer, yet the functional role of SNHG11 in KIRC remains unclear. In this study, we perform integrated multiomics analysis using data from the TCGA and ICGC cohorts and reveal that SNHG11, a methylation-associated lncRNA, is significantly correlated with poor clinical outcomes. In vitro and in vivo assays demonstrate that SNHG11 promotes tumor proliferation and progression. Mechanistically, SNHG11 enhances oxidative phosphorylation, as evidenced by increased ATP production, disrupted mitochondrial membrane potential, and altered NAD+/NADH ratios. Furthermore, SNHG11 expression is associated with somatic mutation patterns, particularly those involving BAP1 and PBRM1, indicating potential crosstalk between epigenetic regulation and genetic alterations. These findings reveal that SNHG11 is a novel biomarker in KIRC and a potential therapeutic target.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2025222
Poultry production faces escalating challenges from intensive farming practices, where stressors, including high stocking density, pathogen exposure, and dietary fluctuations, disrupt intestinal integrity, microbiota balance, and antioxidant defenses. These disruptions impair nutrient absorption, growth performance, and immune function, leading to significant economic losses. Although antibiotics have historically mitigated such issues, growing restrictions due to antimicrobial resistance necessitate natural alternatives. Fungal polysaccharides (FP)—notably lentinan (LNT) from Lentinula edodes and polysaccharide from Ganoderma lucidum (GLP), are promising candidates owing to their immunomodulatory, antioxidant, and prebiotic properties. However, existing research focuses predominantly on individual FP, neglecting potential synergies in blended formulations. Structurally, LNT (β-(1→3)-D-glucan backbone) enhances rumen volatile fatty acid production and fiber degradation, whereas GLP (heterogeneous α/β-glycans) potently activates the Nrf2/HO-1 antioxidant pathway and modulates Th1/Th2 immunity. These divergent mechanisms imply complementary effects when combined. In our previous experiments on broiler feeding, we reported that a combination of GLP (68.32% polysaccharide content, composed of mannose, glucose, arabinose, rhamnose, and galactose at a molar ratio of 1.00:16.37:18.82:1.42:17.42) and LNT (76.52% polysaccharide content, composed of mannose, galacturonic acid, arabinose, galactose, glucose, and rhamnose at a molar ratio of 1.00:15.22:8.23:2.05:1.78:4.26) at a 1:1 ratio maximally promoted broiler growth (unpublished data), but their impacts on intestinal morphology, antioxidant signaling, and the microbiota remain uncharacterized. We therefore hypothesize that mixed FP synergistically may enhance intestinal health by simultaneously improving nutrient absorption, activating antioxidant pathways, and stabilizing microbial ecosystems. To investigate the effects of mixed FP on intestinal development, 240 one-day-old Arbor Acres male broilers were randomly assigned to the 0 mg/kg FP (Control), 200 mg/kg FP (Group I), 400 mg/kg FP (Group II), and 600 mg/kg FP (Group III) groups. Broilers were housed in three-tier battery cages (0.7 m × 0.7 m × 0.4 m; 12 broilers/cage), with five replicate cages per experimental group maintained under identical conditions. The experiments were approved by the College of Animal Science and Technology in Anhui Agricultural University (approval number: SYXK 2016-007). All the cages were subjected to a 16 h light: 8 h dark cycle with ad libitum access to water and twice-daily feeding (09:00/16:00) of basal diets (Supplementary Table S1). On day 42, the duodenum, jejunum, and ileum segments were collected, fixed in 4% paraformaldehyde, sectioned at 5 μm, and stained with hematoxylin-eosin. Villus height (VH), crypt depth (CD), and VH/CD ratios were measured via Case Viewer software. The results revealed that Group II significantly increased VH and VH/CD across all the intestinal segments while reducing CD (Figure 1A; P < 0.05 vs the control); these findings suggest enhanced nutrient absorption capacity and intestinal health. To evaluate antioxidant capacity and signaling pathway activation, intestinal tissues were homogenized in PBS (1:9, w/v). The total antioxidant capacity (T-AOC), total superoxide dismutase (T-SOD), and glutathione peroxidase (GSH-Px) activities were determined via commercial kits (Nanjing Jiancheng Bioengineering Institute, Nanjing, China)). For gene expression analysis, total RNA was extracted and reverse-transcribed. The qPCR was performed via specific primers for HO-1, NQO1, CAT, Nrf2, and Keap1, with β-actin used as the reference gene (primer sequences and product sizes are listed in Supplementary Table S2). The results demonstrated that Group II significantly elevated antioxidant enzyme activities (P < 0.05), upregulated HO-1, NQO1, CAT, and Nrf2, and
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2025048
As a classic tumor suppressor gene, p53 has been extensively studied since its discovery in the mid-1980s. Research findings have revealed that p53 protein expression is suppressed in various cancers [1]. For example, in cervical cancer, p53 predominantly exists in a wild-type form to maintain its biological function [2]. Nevertheless, its tumor-suppressive activity is significantly impaired because of rapid protein degradation, short half-life, and low levels. Post-translational modifications (PTMs) of p53, such as ubiquitination, acetylation, phosphorylation and methylation, are critical regulators of its stability, activity, conformation, localization, and interactions with cofactors [3]. Among these, ubiquitination and acetylation play central roles in controlling p53 protein stability and activity [4]. Therefore, targeting p53 PTMs to modulate its ubiquitination and acetylation levels represents an effective strategy to increase its stability and tumor-suppressive function, offering a promising avenue for cervical cancer drug development. In 99% of cervical cancers (high-risk human papillomavirus-positive), E3 ubiquitin ligase E6-associated protein (E6AP) mediates the ubiquitination degradation of p53 [5], whereas histone deacetylase 6 (HDAC6) deacetylates p53. In this study, we explored the possibility of combining the natural product withferin A (WA) with the HDAC6 inhibitor ricolinostat (RIC) to treat cervical cancer cells, with a focus on the ubiquitination and acetylation of p53 and the consequences for its stability. These results suggested that the combination of WA and RIC is more effective than either treatment alone in inhibiting the degradation and increasing the stability of p53, thereby synergistically slowing the onset and progression of cervical cancer.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2025233
This is a corrigendum to the article 'Caveolin-1-deficient fibroblasts promote migration, invasion, and stemness by activating the TGF-β/Smad signaling pathway in breast cancer cells' published in Acta Biochim Biophys Sin 54: 1587–1598. The authors identified inaccuracies in the preparation of several figures (Figure 2D, 4A, and 5A) and have replaced them with corrected versions. The errors are strictly confined to figure presentation and do not impact the underlying data, statistical analysis, or main conclusions. The authors apologize for the oversight.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2025047
Systemic lupus erythematosus (SLE) is an autoimmune disease characterized by a complex pathogenesis that was previously thought to involve primarily adaptive immunity. Emerging evidence underscores the role of neutrophils in shaping immune dysregulation and inducing organ damage in lupus. This study aims to investigate the dynamics of neutrophil senescence and its relationship with lupus, an area that remains poorly understood. Here, we identify a significantly elevated proportion of CXCR4hiCD62Llo senescence-like neutrophils in the peripheral blood of SLE patients compare to that in the healthy donors. Increased numbers of senescence-like neutrophils are positively correlated with SLE disease activity and autoantibody production in SLE patients. In addition, senescence-like neutrophils derived from SLE patients exhibit an impaired ability to suppress the proinflammatory activity of natural killer (NK) cells and CD4+ T cells. Further mechanistic exploration suggests that these senescence-like neutrophils might exert their immunosuppressive effects via reactive oxygen species (ROS) production under physiological conditions. Our results demonstrate that senescence-like neutrophils could serve as biomarkers for assessing the disease activity of SLE. The compromised immunosuppressive function of senescence-like neutrophils provides a new perspective on SLE pathophysiology and may pave the way for the development of novel therapies.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2025094
Kainate receptors (KARs) are one of the ionotropic glutamate receptor (iGluR) families, and their antagonists are being investigated for the treatment of several neurological disorders, including Alzheimer’s disease, a neurodegenerative condition, etc. As early as 1990, Bettler et al. [1] first cloned the GRIK1 subunit of KARs, marking a pivotal advancement in understanding these receptors. Members of the iGluR family have been identified in other jawed vertebrates and exhibit conserved structural features. However, research into iGluRs in jawless vertebrates has been limited. Owing to the unique evolutionary position of lampreys, their iGluRs might also present functions distinct from those of jawed vertebrates; therefore, it is particularly important to study iGluRs in lampreys. In this study, we identified four homologous subunits of iGluRs in lampreys, including Lr-GRIA2, Lr-GRIA4, Lr-GRIK1 and Lr-GRIN2B. Lampreys occupy a unique evolutionary position, making phylogenetic analysis of iGluR subunits between lampreys and other species essential for understanding iGluR evolution. Given the distinctive functional characteristics of iGluR family members, particularly KAR subtypes, we focused on the functional validation of Lr-GRIK1. First, we confirmed the expression of Lr-GRIK1 in lampreys and examined its expression profiles across various tissues via qPCR and western blotting. To elucidate the functional role of Lr-GRIK1 in lampreys, we used an siRNA to silence Lr-GRIK1. We subsequently conducted transcriptome sequencing of both the silenced and control groups to construct and analyze their expression profiles. Our analysis revealed differential expression of genes enriched in pathways related to signal transduction and the immune system, highlighting potential roles of Lr-GRIK1 beyond traditional neurotransmission functions. Unlike in jawed vertebrates, transcriptome enrichment provides a new direction for understanding the function of Lr-GRIK1. Therefore, we monitored the changes in Lr-GRIK1 expression in the kidney tissue of lampreys after stimulation. In addition, we confirmed that Lr-GRIK1 affects the expression levels of immune-related molecules during the immune response process. These findings provide insights into the broader functional significance of Lr-GRIK1 in the biology of lampreys.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2025005
Pulpitis is a common inflammatory oral disease that can lead to pulp necrosis. The aim of this study is to investigate the expression and regulatory mechanisms of ATF3, a potential therapeutic marker, in pulpitis. A mouse pulpitis model with different degrees of inflammation is established, and the expression of ATF3 in pulpitis is explored. The histological features of healthy pulp and pulpitis are analyzed by HE staining, and classical inflammatory factors are detected by immunohistochemistry (IHC). In an in vitro study, we investigate the role of ATF3 in the regulation of WNT4 transcription and explore the effects of the ATF3/WNT4 axis on the polarization of RAW264.7 macrophages, the inflammatory response and the osteogenic differentiation of human dental pulp stem/stromal cells (hDPSCs). Our results show that ATF3 is expressed at low levels in inflamed pulp tissues; overexpression of ATF3 reduces the area of pulp necrosis, decreases the level of pro-inflammatory factors, and promotes macrophage polarization toward the M2 type. Furthermore, we reveal that ATF3 binds to the WNT4 promoter region and positively regulates the expression of WNT4 and that ATF3 downregulates M1 markers and increases the expression of M2 markers by regulating WNT4 expression. In addition, ATF3 promotes the osteogenic differentiation of dental pulp stem cells. In summary, this study reveals that ATF3 promotes M2 macrophage polarization by regulating WNT4, which in turn inhibits pulpal inflammatory responses and promotes the osteogenic differentiation of dental pulp stem cells. These findings suggest that ATF3 may be a potential target for pulpitis treatment.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2024239
Reactivating the embryonic ζ-globin gene represents a potential therapeutic approach to ameliorate the severe clinical phenotype of α-thalassemia and sickle cell disease. The transcription factor MYB has been extensively proven to be a master regulator of the γ-globin gene, but its role in the regulation of ζ-globin remains incompletely understood. Here, we report a mechanistic study on the derepression of ζ-globin both in vivo and in vitro. We show that MYB depletion in mouse models and human hematopoietic stem cells leads to consistent and remarkable reactivation of ζ-globin. Furthermore, multiomics analysis and functional validation of MYB-knockout and wild-type cell lines reveal that ETO2 functions as a novel repressor of ζ-globin through coordination with NuRD nucleosome remodeling and the deacetylation complex to modulate histone deacetylation of ζ-globin. Additionally, we evaluate the clinical significance of these findings by knocking out ETO2 in primary CD34+ cells from nondeletional hemoglobin H patients, which results in a significant increase in ζ-globin expression. The RNA-seq data reveal that key erythroid genes are more co-regulated by Myb and Eto2 than by Myb and Klf1, highlighting a distinctly enhanced erythroid-specific transcriptional impact within the MYB-ETO2 regulatory axis. Compared with ETO2 knockout alone, codepletion of ETO2 and BCL11A did not significantly activate ζ-globin, suggesting that the MYB-ETO2 pathway primarily silences ζ-globin. Our study reveals a linear MYB-ETO2 signaling pathway crucial for ζ-globin repression and offers new targets for treating α-thalassemia and sickle cell disease.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2025165
Sepsis-induced cardiomyopathy (SIC) is a severe complication of sepsis and septic shock and is characterized by cardiac dysfunction. Levosimendan (LEVO), a calcium sensitizer, has shown therapeutic potential in SIC, although its underlying mechanism remains unclear. Nrf2, a pivotal regulator of antioxidant and anti-inflammatory responses, may represent a potential target for SIC treatment. In this study, we examine the effects of LEVO on SIC and explore the mechanistic role of Nrf2 in mediating its cardioprotective effects. A murine SIC model is established via cecal ligation and puncture (CLP), and cardiomyocyte injury is induced in vitro via lipopolysaccharide (LPS) exposure in HL-1 cells. The CLP procedure significantly elevates serum cTnI and IL-6 levels and reduces the survival rates of mice. Echocardiographic analysis reveals impaired cardiac structure and function, accompanied by mitochondrial morphological and functional damage, in SIC mice. Interestingly, these pathological changes in SIC are markedly attenuated by LEVO treatment. Similarly, LEVO administration restores proliferative capacity; increases mitochondrial ATP, mitochondrial membrane potential (MMP) and NADH levels; and reduces ROS production and intracellular calcium overload. Notably, the protective effects of LEVO on cardiomyocyte viability and mitochondrial function are significantly diminished following Nrf2 inhibition or Nrf2 knockout (KO). Collectively, these findings demonstrate that LEVO mitigates cardiomyocyte injury and mitochondrial dysfunction in SIC through an Nrf2-dependent mechanism.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2025178
Asthma is a prevalent chronic respiratory disease in children. Recently, adjusting the Th1/Th2 imbalance has become a significant focus in asthma immunotherapy. The present study aims to investigate the roles and mechanisms of circDCBLD2 in maintaining the Th1/Th2 immune balance. CircDCBLD2 is downregulated in CD4+ T cells from asthmatic patients and in CD4+ T cells from an OVA-induced asthmatic mouse model. Additionally, circDCBLD2 levels are significantly decreased in the PBMCs of asthmatic mice. The expression of circDCBLD2 is positively correlated with the Th1 cytokines IFN-γ and IL-2 but negatively correlated with the Th2 cytokines IL-4 and IL-13. Flow cytometry and ELISA analyses demonstrate that circDCBLD2 overexpression increases the proportion of Th1 cells (CD4+IFN-γ+) and the levels of Th1 cytokines while decreasing the proportion of Th2 cells (CD4+IL-4+) and the levels of Th2 cytokines. Furthermore, circDCBLD2 overexpression alleviates the asthma phenotype in OVA-induced mice, reduces the infiltration of inflammatory cells in the lungs, and corrects the Th1/Th2 imbalance. Mechanistically, circDCBLD2 is found to target miR-26a-5p. Rescue experiments indicate that circDCBLD2 regulates the Th1/Th2 immune balance by targeting miR-26a-5p. Additionally, PTEN has been identified as a direct target of miR-26a-5p. The overexpression of PTEN partially reverses the effects of miR-26a-5p on the Th1/Th2 immune balance. These findings indicate that circDCBLD2 increases the proportion of Th1 cells and decreases the proportion of Th2 cells via the miR-26a-5p/PTEN axis, providing a promising target for asthma treatment.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2025011
Relieving hypoxia in the tumor microenvironment (TME) promotes innate and adaptive immunity. Our previous research demonstrated that reoxygenation of the TME promotes the phagocytosis and tumor-killing functions of tumor-associated macrophages (TAMs) by upregulating pyruvate carboxylase (PCB). However, the mechanism remains obscure. In the present study, we find that versican derived from melanoma cells binds to TLR2 and activates the downstream transcription factor RelB, which transcribes PCB under normoxia. Blocking the versican-TLR2-MyD88-RelB axis not only reverses the upregulation of PCB in TAMs but also hinders the clearance of tumor cells by TAMs. Our work suggests a pathway that modulates the functions of TAMs under normoxia, which could be harnessed for strengthening anti-tumor immunity.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2024228
Annexins are a family of multifunctional calcium-dependent and phospholipid-binding proteins that are widely distributed in the plant kingdom. They have a highly conserved evolutionary history that dates back to single-celled protists. Plant annexins, as soluble proteins, can flexibly bind to endomembranes and plasma membranes, exhibiting unique calcium-dependent and calcium-independent characteristics. Members of the annexin family have diverse functions, including binding to F-actin, participating in ATP and GTP hydrolysis, and even serving as peroxidases or cation channels. Annexins play pivotal roles in plant growth and stress signaling. They can respond sensitively to environmental, metabolic, and developmental signals, thereby affecting cytoskeleton remodeling and exocytosis mechanisms. Plant annexin gene families have been successfully identified in multiple species, and their expression and intracellular localization are precisely regulated by developmental processes and environmental factors. Although research on plant annexins has aroused great interest, their depth and breadth still need further expansion compared with those of animal annexins. This article provides a comprehensive and in-depth review of the characteristics and functions of plant annexin families, revealing their core roles in plant growth and adaptation, and yielding valuable references and insights for future research.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2025146
Actin-bundling protein Fascin1 (FSCN1) is encoded by the Fscn1 gene and is crucial for cytoskeletal remodeling and cellular migration. Although a previous study linked Fscn1 deficiency to neonatal lethality in mice, the underlying metabolic mechanism remains unclear. In this study, we report that systemic knockout (KO) of Fscn1 leads to 52.2% mortality within 24 h post-birth, accompanied by severe hypoglycemia in KO pups compared with their littermates. Remarkably, this lethality is fully rescued by oral glucose administration, indicating a glucose supply-dependent survival mechanism. Surviving Fscn1-KO neonates display persistent developmental deficits, including growth retardation and depleted lipid stores, despite intact canonical insulin-regulated hepatic gluconeogenic pathways. Transcriptomic profiling of P0 livers reveals that Fscn1 loss predominantly disrupts metabolic pathways, with the glycerol phosphate shuttle being the most significantly downregulated module. Mechanistically, Fscn1-KO livers exhibit markedly reduced protein levels of glycerol-3-phosphate dehydrogenase isoforms (GPD1/GPD2), key enzymes bridging glycolysis and gluconeogenesis. Consistently, glycerol tolerance tests demonstrate impaired glycerol-to-glucose conversion in Fscn1-KO mice, confirming defective glycerol-driven gluconeogenesis. Our findings establish FSCN1 as a novel cytoskeletal-metabolic integrator essential for neonatal survival by sustaining hepatic glucose production from glycerol, thus revealing an unexpected role of actin dynamics in coordinating metabolic adaptation during early postnatal development.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2025130
Tumor radioresistance and severe toxicity make reirradiation for recurrent nasopharyngeal carcinoma (NPC) a significant clinical challenge. This study aims to investigate the ability of the poly(ADP-ribose) polymerase (PARP) inhibitor olaparib to sensitize recurrent NPC cells irradiated with photon or carbon ion (C-ion), and to explore the underlying mechanism of the synergistic promotion of cell death by olaparib and ionizing radiation. The results show that olaparib has significant X-ray and C-ion radiosensitization effects on recurrent NPC cells and the associated HK-RR photon-resistant model. Radiation, particularly C-ion exposure, induces a homologous recombination (HR)-deficient gene signature in HR-proficient NPC cells, potentially increasing their sensitivity to PARP inhibition. C-ion and X-ray irradiation induces similar modes of cell death, and multiple cell death pathways [including apoptosis, necrosis, ferroptosis, senescence, and autophagic cell death (ACD)] contribute to the cytotoxic effects of radiation combined with olaparib, with ACD being the dominant pathway. Both the pharmacological and genetic inhibition of autophagy significantly attenuate the radiosensitization effect of olaparib. In conclusion, olaparib effectively sensitizes recurrent NPC cells to both X-ray irradiation and C-ion irradiation, with autophagy playing a central role in mediating this effect.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2025237
Exposure to chronic hypobaric hypoxia provokes marked alterations in the gut microbiota and its metabolome, yet the functional significance of histidine-derived metabolites in hypobaric hypoxic pulmonary hypertension (PH) remains underexplored. Here, we employ 16S rDNA, metagenomic, and untargeted metabolomic sequencing to characterize longitudinal shifts in the fecal microbiota and metabolites during hypobaric hypoxic PH development in Sprague-Dawley rats. Fecal carnosine levels and the abundance of its producer, Ruminococcus bromii, both decrease significantly over 28 days of hypobaric hypoxia (P < 0.05). Spearman correlation shows that carnosine is inversely correlated with the percentage of pulmonary arteriole media thickness (MT%; r = –0.8741, P < 0.001). Therapeutic supplementation with carnosine restores systemic and pulmonary antioxidant defenses and attenuates vascular remodeling without altering right ventricular pressures. In vitro, carnosine inhibits hypoxia-induced pulmonary artery smooth muscle cell (PASMC) proliferation and migration and suppresses nuclear factor erythroid 2-related factor 2 (Nrf2) accumulation. These findings reveal dynamic gut-lung crosstalk in hypobaric hypoxic PH and nominate carnosine as a metabolite-based intervention to mitigate hypoxia-driven pulmonary vascular remodeling.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2026027
Phosphorylated CTD Interacting Factor 1 (PCIF1) is the exclusive methyltransferase responsible for the N6,2-O-dimethyladenosine (m6Am) modification in mammalian mRNA. Our previous research identified PCIF1 as a potent tumor suppressor in glioma, demonstrating its ability to impair cell proliferation, induce G2/M phase arrest, and promote apoptosis. However, its role in glioma cell migration and invasion remains unclear. In this study, we investigate how PCIF1 regulates glioma cell migration and invasion. Overexpression of PCIF1 inhibits migration and invasion, whereas PCIF1 knockdown enhances these behaviors. Corresponding changes are observed in mesenchymal markers (Vimentin, β-catenin, Snail, Slug) and the epithelial marker T-cadherin, indicating that PCIF1 suppresses epithelial-to-mesenchymal transition (EMT)-mediated glioma invasion. Mechanistically, PCIF1 modulates the AKT pathway by promoting proteasomal degradation of AKT while increasing phosphorylated AKT (p-AKT) levels, revealing a complex regulatory mechanism. PCIF1 knockdown upregulates INPP5B, a lipid phosphatase, causing accumulation of PI(3,4)P2 and enhanced AKT activation. Conversely, PCIF1 overexpression increases PI(3,4,5)P3 production, elevating p-AKT levels. This bidirectional regulation suggests that PCIF1 influences phosphoinositide signaling and AKT activation. Our findings highlight PCIF1 as a key modulator of glioblastoma cell migration and invasion through phosphoinositide signaling, positioning it as a potential biomarker and therapeutic target in glioma.
Acta Biochimica et Biophysica Sinica•2024•DOI: 10.3724/abbs.2024148
Ubiquitin-like containing PHD and RING finger domains 1 (UHRF1) is involved in tumorigenicity through DNA methylation in various cancers, including breast cancer. This study aims to investigate the regulatory mechanisms of UHRF1 in breast cancer progression. Herein, we show that UHRF1 is upregulated in breast cancer tissues and cell lines as measured by western blot analysis and immunohistochemistry. Breast cancer cells are transfected with a UHRF1 overexpression plasmid (pcDNA-UHRF1) or short hairpin RNA targeting UHRF1 (sh-UHRF1), followed by detection of cell proliferation, invasion, apoptosis, and cell cycle. UHRF1 overexpression promotes proliferation and invasion and attenuates cell cycle arrest and apoptosis in breast cancer cells, while UHRF1 knockdown shows the opposite effect. Moreover, methylation-specific PCR and ChIP assays indicate that UHRF1 inhibits zinc finger and BTB domain containing 16 (ZBTB16) expression by promoting ZBTB16 promoter methylation via the recruitment of DNA methyltransferase 1 (DNMT1). Then, a co-IP assay is used to verify the interaction between ZBTB16 and the annexin A7 (ANXA7) protein. ZBTB16 promotes ANXA7 expression and subsequently inhibits Cyclin B1 expression. Rescue experiments reveal that ZBTB16 knockdown reverses the inhibitory effects of UHRF1 knockdown on breast cancer cell malignancies and that ANXA7 knockdown abolishes the inhibitory effects of ZBTB16 overexpression on breast cancer cell malignancies. Additionally, UHRF1 knockdown significantly inhibits xenograft tumor growth in vivo. In conclusion, UHRF1 knockdown inhibits proliferation and invasion, induces cell cycle arrest and apoptosis in breast cancer cells via the ZBTB16/ANXA7/Cyclin B1 axis, and reduces xenograft tumor growth in vivo.
Acta Biochimica et Biophysica Sinica•2024•DOI: 10.3724/abbs.2024114
With the increasing global population and aging demographic, the incidence of stroke is rising. Among these, ischemic stroke (IS), also known as cerebral ischemia, constitutes over 80% of all stroke cases. This condition is characterized by an acute cerebrovascular disease caused by the blockage and interruption of the brain's blood supply, resulting in localized tissue ischemia, oxygen, and glucose deficiency, ultimately leading to the death of nerve cells and tissue necrosis [1,2]. "Vascular recanalization and the restoration of cerebral blood flow" are the primary clinical treatment objectives and are achieved through the intravenous administration of drugs such as tissue plasminogen activator or through surgical thrombectomy. These interventions not only restore the delivery of oxygen and glucose to the affected cerebral area but also help prevent the expansion of the infarcted region. However, the restoration of reperfusion cerebral blood flow similarly exposes the infarct area to peripheral immune cells, triggering the activation of the immune response and inflammation-induced injury [3]. Research indicates that IS elicits a robust inflammatory response, with neuroinflammation playing a crucial role in the secondary neurodegeneration process following stroke. Neuroinflammatory responses are initiated and perpetuated through injury cascades that include the release of inflammatory mediators, the migration and recruitment of white blood cells across the blood-brain barrier, and the impairment of endothelial nitric oxide synthase. These mechanisms collectively promote the activation of pro-inflammatory genes, which in turn activate microglia (MG) and exacerbate ischemic damage and neurological dysfunction [4]. MG are resident immune cells of the central nervous system (CNS). Its function is akin to that of macrophages, serving as the first line of defense against injuries within the central nervous system. Under typical conditions, brain microglia participate in immune surveillance and defense against infectious agents. However, in the pathogenesis of neurodegenerative diseases such as IS, MG are activated by various stimuli. Once activated, MG are known to release numerous proinflammatory or cytotoxic factors, such as inducible nitric oxide synthase (iNOS), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and reactive oxygen species (ROS). These factors initiate the neuroinflammatory response, exacerbate inflammation, intensify damage to brain tissue and neurons, and significantly hinder the repair of brain injuries and neurogenesis [5,6]. Therefore, inhibiting the activation of microglia and reducing the inflammatory response in the central nervous system are crucial for minimizing brain damage caused by IS and are vital for developing effective prevention and treatment strategies. In recent years, certain natural compounds extracted from traditional drug formulations have shown high therapeutic potential in protecting the brain from cerebral ischemic injury. These compounds reduce the neuroinflammatory response and apoptosis following stroke. Traditional Chinese herbal medicine (TCHM) and its constituent herbs feature a multiplicity of components, targets, and pathways owing to their complex formulations and therapeutic principles, making them promising sources for developing effective treatments for IS. Panax notoginseng saponin (PNS), as the principal bioactive component of Panax notoginseng, is extensively utilized in the prevention and treatment of cardiovascular and cerebrovascular diseases. Its pharmacological benefits include dissipating blood stasis, promoting hemostasis, alleviating swelling and pain, regulating energy metabolism disorders, balancing ion metabolism, and reducing and accelerating the clearance of free radicals [7]. Research indicates that PNS mitigates apoptosis by maintaining mitochondrial homeostasis, enhancing the integrity of the blood‒brain barrier (BBB), augmenting cerebral blood supply, and fostering the differentiation of neural stem cells and proliferation of hippocampal neurons. In addition, PNS offers neuroprotection against focal cerebral I/R injury in rats by reducing brain edema, upregulating the expression of the heat shock protein HSP70, and downregulating the expression of transferrin [8,9]. Additionally, PNS has been reported to enhance the recovery of neurogenesis and neurological function in cerebral embolism induced by microspheres and to reduce sepsis-induced acute kidney injury by suppressing inflammation [10]. However, the mechanism by which PNS targets IS has not been fully elucidated. In this study, we investigated the anti-inflammatory effects of PNS on IS and identified potential target pathways that could inhibit microglia-mediated inflammatory response.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2025141
Colorectal cancer (CRC) is a common type of gastrointestinal malignancy, and it has a close connection with long noncoding RNAs (lncRNAs). This study aims to examine the involvement of long noncoding RNA LINC00114, which targets heterogeneous nuclear ribonucleoprotein A1 (HNRNPA1) in regulating glutamine metabolism and angiogenesis in the metastasis of colorectal cancer (CRC). LINC00114 and HNRNPA1 levels are measured in CRC tissues and cells to determine their expression levels. Then, siRNA targeting LINC00114 (si-LINC00114) is used to transfect CRC cells, and cell proliferation and metastasis are detected. The influence of exogenous glucose and glutamine supplementation on angiogenesis induced by LINC00114 in CRC is investigated in HUVECs. Glutamine metabolism in CRC cells is also detected. Furthermore, the role of LINC00114 in CRC xenograft tumors is studied in vivo. LINC00114 and HNRNPA1 are highly expressed in CRC and positively correlate with CD31. si-LINC00114 significantly inhibits proliferation, metastasis and HNRNPA1 expression in CRC cells. An RNA-binding-protein immunoprecipitation (RIP) assay confirms that LINC00114 can bind to HNRNPA1 and positively regulate its expression. Further experiments confirm that si-LINC00114 significantly inhibits cell proliferation and tubule formation in HUVECs. Exogenous glucose and glutamine supplementation significantly promotes the levels of LINC00114 and HNRNPA1 in CRC cells and promotes tubule formation in HUVECs. In addition, transfection of CRC cells with si-LINC00114 and/or oe-HNRNPA1 regulates glutamine metabolism in CRC cells. Animal studies confirm that intervention with LINC00114 represses the progression and vascular normalization of CRC and regulates glutamine metabolism. In conclusion, LINC00114 promotes CRC metastasis by targeting HNRNPA1 to regulate glutamine metabolic reprogramming and angiogenesis.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2025069
Inflammation, while essential for host defense, can contribute to diseases like inflammatory bowel disease (IBD) when dysregulated. IBD involves chronic intestinal inflammation linked to bacterial infection and disruption of the intestinal epithelial barrier (IEB). The IEB maintains intestinal homeostasis, preventing the entry of harmful substances like lipopolysaccharide (LPS) while allowing nutrient absorption. Preserving IEB function is crucial in managing various diseases. Current IBD treatments often have adverse effects, including facial swelling and acne from corticosteroids, and allergic reactions, nausea, elevated liver tests, and pancreatitis from immunomodulators, increasing interest in natural products. Flavonoids, a diverse class of plant metabolites, have shown promising anti-inflammatory properties. Lutonarin (LU), a flavonoid abundant in barley seedlings, has exhibited various biological activities, including anti-inflammatory effects in LPS-stimulated macrophages. However, its specific impacts on IEB function remain unclear. This study aims to investigate the protective effects of LU against LPS-induced IEB dysfunction using an in vitro Caco-2 cell monolayer model. To assess the possible cytotoxicity of LU and optimize the suitable concentration, the viability of Caco-2 cells was assessed by CCK-8 assay. LU concentrations up to 96 μM did not significantly affect cell viability after 24 and 48 h of exposure. To evaluate the protective effect of LU against LPS-induced cytotoxicity, Caco-2 cells were pretreated with LU (3‒96 μM) for 48 h prior to exposure to 15 μg/mL LPS for 24 h. LPS significantly reduced cell viability, while LU pretreatment attenuated this reduction in a concentration-dependent manner. Concentrations of 12 μM LU were selected for subsequent experiments to minimize potential off-target effects. The Caco-2 cell monolayer model, a well-established in vitro system for investigating IEB function, was employed to study the effects of LU on IEB integrity. Barrier integrity was evaluated using transepithelial electrical resistance (TEER) measurements. LPS treatment significantly reduced TEER, indicating impaired barrier function. Pretreatment with 12 μM LU preserved TEER values, suggesting a protective effect against LPS-induced barrier disruption. Furthermore, paracellular permeability was evaluated using fluorescein isothiocyanate-dextran 4 (FITC-dextran, 4 kDa). LPS significantly increased FITC-dextran (FD4) flux, indicating increased permeability. LU pretreatment markedly attenuated this effect, confirming its ability to prevent LPS-induced permeability changes. To detect if LPS and LU pretreatment changes the expressions of tight junction (TJ) proteins Zonula occludens-1 (ZO-1) and Occludin, real-time qPCR and immunofluorescence staining assays were applied. LPS treatment significantly reduced mRNA expression levels of TJ proteins ZO-1 and Occludin. LU pretreatment effectively mitigated this downregulation, restoring their expression to levels comparable to the CON group.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2024213
Approximately one-tenth of the global population is affected by diabetes mellitus, and its incidence continues to rise each year. In China, 1.4 million patients die of diabetes-related complications every year. Additionally, approximately 26% of patients with diabetes develop diabetic cardiomyopathy, with heart failure being one of the main causes of death in these patients. However, early detection of diabetic cardiomyopathy has proven to be difficult in a clinical setting; furthermore, there are limited guidelines and targeted means of prevention and treatment for this disease. In recent years, several studies have provided evidence for the occurrence of various forms of regulated cell death in diabetic myocardial cells, including apoptosis, necroptosis, ferroptosis, and cuproptosis, which are closely linked to the pathological progression of diabetic cardiomyopathy. Although most research on diabetic cardiomyopathy is currently in the animal trial phase, the inhibition of these regulatory cell death processes can limit or slow down the progression of diabetic cardiomyopathy. Therefore, this review discusses the appropriate animal experimental models currently available for diabetic cardiomyopathy and evaluates the roles of apoptosis, necroptosis, ferroptosis, and cuproptosis in diabetic cardiomyopathy. We hope to provide new methods and ideas for future research in diabetic cardiomyopathy.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2024157
The biomolecular assemblies (condensates or aggregates) formed by mutant proteins are a pathological hallmark of neurodegenerative diseases. Some RNA-binding proteins (RBPs) are typically prone to aggregation that is closely associated with disease pathologies. These RBPs include numerous well-recognized pathogenic proteins, such as TAR DNA binding protein of 43 kDa (TDP-43), fused in sarcoma (FUS), ataxin-2 (Atx2), and poly(A)-binding protein nuclear 1 (PABPN1). Recent studies have revealed that liquid-liquid phase separation (LLPS), as a mechanism, underlies the highly dynamic and reversible granule formation of RBPs, and highlighted that multivalent RNA molecules play crucial roles in this process. These granules are necessary for diverse physiological functions, such as RNA splicing, trafficking, and even RNA storage, during stress. However, the aberrant phase transition of these mutant RBPs usually results in the formation of solid-like aggregates or inclusions within both the cytoplasm and nucleus. More importantly, aggregates formed by RBPs can sequester specific proteins, RNAs or other interacting partners, consequently contributing to RBP-related pathologies. For example, wild-type PABPN1 forms dynamic nuclear speckles with the assistance of poly(A) RNAs, whereas Ala expansion of PABPN1 results in the formation of aggregates, which are involved in the disease progression of oculopharyngeal muscular dystrophy (OPMD). Although the biological importance of various RBP granules is realized in either the cytoplasm or nucleus, how RNA regulates the formation of granules and the transition to aberrant RBP aggregates remains largely unknown. The interaction of a protein with other biomolecules (proteins, nucleic acids, etc.) is the prerequisite for the protein executing its normal biological function in cells. Identifying protein-protein and protein-RNA interactions is fundamental for the biochemical investigation of an individual protein and for attempts to understand the functional role of the protein. To date, many methods for studying protein-protein interactions have been developed on the basis of various principles, but it is still difficult to clarify whether the interactions between proteins, especially the RBPs involved, are direct or indirect, since RBPs generally bind to diverse RNAs closely and are incorporated into macromolecular ribonucleoprotein (RNP) complexes. We have taken several pairs of RBPs as examples, including TDP-35 (C-terminal 35-kDa fragment of TDP-43) with TDP-43 or TIA1, PABPN1 with a 25-kDa component of the mammalian cleavage factor I complex (CFIm25) and Atx2 with DEAD-box RNA helicase 6 (DDX6), and applied modified co-immunoprecipitation (Co-IP) and supernatant/pellet (S/P) fractionation experiments to characterize the association and sequestration of RBPs by using single-stranded DNA (ssDNA) chimera under ribonuclease (RNase) treatment. We designed several pieces of ssDNA oligonucleotides to mimic particular RNAs in cells that may mediate the association and sequestration of RBPs. The association of RBP proteins generally requires binding with multivalent RNA chains, since the bound RNAs tend to incorporate into a large protein-RNA complex with the help of RNA molecules. In Co-IP assay, especially for RBPs, RNase is often utilized to digest RNA in cell lysates to characterize whether the association of different RBPs is direct or indirect. It is important for us to demonstrate the active role of particular RNAs in the association or interaction of RBPs. Therefore, we designed and synthesized ssDNA chimeras to mimic the corresponding RNA that specifically bind to both RBPs simultaneously. In this case, ssDNA is used for rescuing the association of RBPs under the condition of RNase treatment, since the ssDNA oligonucleotide is resistant to nuclease activity. To design ssDNA chimeras for the RBPs of interest, first, the RNA sequences that bind to the two RPBs should be defined. The ssDNA should contain at least two portions (motifs) that specifically bind to each RBP, and each ssDNA portion may include 2–3 repeats of the binding sequence, so that the ssDNA can be recognized and bound efficiently by each RBP. Notably, the T base in ssDNA may sometimes be replaced with the U base (dU) for some more specific-binding RBPs, such as PABPN1. In the case of TDP-43 with Atx2, the binding specificities of the RNA sequences for TDP-43 and Atx2 are UG-rich and AUUUUU (AU5), respectively; then, the TG repeat portion is designed to bind to TDP-43, and the AT5 repeat is to bind to Atx2. Thus, an integrated method of co-IP and S/P fractionation was applied to characterize the association and sequestration of RBPs by combining ribonuclease (RNase) and ssDNA treatments.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2024237
The interaction between TF binding and DNA methylation is increasingly recognized as a key player in the regulation of gene expression. However, the role of this interaction in regulating ICAM1 expression in breast cancer has not been elucidated. CpG methylation in the ICAM1 promoter is negatively correlated with ICAM1 expression, and ICAM1 expression is significantly positively correlated with DNMT and TET3 expression in breast cancer. TF binding attenuates ICAM1 promoter CpG methylation and promotes ICAM1 transcription. DNA methylation regulation enhances ICAM1 expression in breast cancer by promoting the transcription of transcription factors. In terms of mechanisms, RELA and STATs recruit TET3 to prevent DNMT-mediated DNA methylation, thereby maintaining CpG island hypomethylation in the ICAM1 promoter. Therefore, TF occupancy limits DNA methylation and affects ICAM1 expression in breast cancer.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2025153
Mitochondrial dysfunction critically disrupts adipocyte remodeling by impairing the thermogenic browning process essential for combating obesity through the upregulation of uncoupling protein 1 (UCP1) and mitochondrial biogenesis. Deficiencies in mitochondrial metabolism, dynamics (including fusion/fission), and autophagy suppress adipocyte plasticity, directly inhibiting UCP1 expression and destabilizing the PPAR-γ/PGC-1α and adenosine 5′-monophosphate (AMP)-activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR) signaling pathways. These disruptions reduce energy expenditure, exacerbate insulin resistance, and promote metabolic syndrome. Moreover, mitochondrial inactivation intersects with neurodegenerative disorders via oxidative stress induced by β-amyloid and α-synuclein aggregation, amplifying systemic metabolic dysregulation. Structural mitochondrial anomalies further impede lipid utilization and adipose tissue adaptation, but unresolved crosstalk between mtDNA and nuclear DNA complicates therapeutic targeting. Future research must prioritize spatiotemporal mapping of mitochondrial dynamics in adipocyte differentiation via single-cell omics to identify key regulatory nodes. Addressing these mechanisms could unlock precision therapies, such as gene editing, to restore mitochondrial function, enhance adipocyte browning, and mitigate obesity, related pathologies alongside neurodegenerative and age-associated diseases.
Acta Biochimica et Biophysica Sinica•2024•DOI: 10.3724/abbs.2023264
Cellular senescence is an important factor leading to pulmonary fibrosis. Deficiency of 8-oxoguanine DNA glycosylase (OGG1) in mice leads to alleviation of bleomycin (BLM)-induced mouse pulmonary fibrosis, and inhibition of the OGG1 enzyme reduces the epithelial mesenchymal transition (EMT) in lung cells. In the present study, we find decreased expression of OGG1 in aged mice and BLM-induced cell senescence. In addition, a decrease in OGG1 expression results in cell senescence, such as increases in the percentage of SA-β-gal-positive cells, and in the p21 and p-H2AX protein levels in response to BLM in lung cells. Furthermore, OGG1 promotes cell transformation in A549 cells in the presence of BLM. We also find that OGG1 siRNA impedes cell cycle progression and inhibits the levels of telomerase reverse transcriptase (TERT) and LaminB1 in BLM-treated lung cells. The increase in OGG1 expression results in the opposite phenomenon. The mRNA levels of senescence-associated secretory phenotype (SASP) components, including IL-1α, IL-1β, IL-6, IL-8, CXCL1/CXCL2, and MMP-3, in the absence of OGG1 are obviously increased in A549 cells treated with BLM. Interestingly, we demonstrate that OGG1 binds to p53 to inhibit the activation of p53 and that silencing of p53 reverses the inhibition of OGG1 on senescence in lung cells. Additionally, the augmented cell senescence is shown in vivo in OGG1-deficient mice. Overall, we provide direct evidence in vivo and in vitro that OGG1 plays an important role in protecting tissue cells against aging associated with the p53 pathway.
Acta Biochimica et Biophysica Sinica•2024•DOI: 10.3724/abbs.2023253
The ovarian surface epithelium (OSE) is a single layer of squamous-to-cuboidal epithelial cells that experience repetitive ovulatory rupture and subsequent repair. However, the characteristics of human immortalized ovarian surface epithelial cells (IOSE80) remain elusive. This study aims to determine whether IOSE80 cells have the characteristics of stem cell proliferation and multilineage differentiation and their application in regenerative medicine. IOSE80 cells are sequenced by high-throughput transcriptome analysis, and 5 sets of public data are used to compare the differences between IOSE80 cells and bone marrow mesenchymal stem cells, pluripotent stem cells, and oocytes in transcriptome profiling. The IOSE80 cells present a cobblestone-like monolayer and express the epithelial cell marker KRT18; the stem cell markers IFITM3, ALDH1A1, and VIM; lowly express stem cell marker LGR5 and germ cell markers DDX4 and DAZL. In addition, the GO terms “regulation of stem cell proliferation”, “epithelial cell proliferation”, etc., are significantly enriched (P<0.05). IOSE80 cells have the potential to act as mesenchymal stem cells to differentiate into adipocytes with lipid droplets, osteoblasts, and chondroblasts in vitro. IOSE80 cells express pluripotent stem cell markers, including OCT4, SSEA4, TRA-1-60, and TRA-1-81, and they can be induced into three germ layers in vitro. IOSE80 cells also form oocyte-like cells in vitro and in vivo. In addition, IOSE80 cells exhibit robust proliferation, migration, and ovarian repair functions after in vivo transplantation. This study demonstrates that IOSE80 cells have the characteristics of pluripotent/multipotent stem cells, indicating their important role in tissue engineering and regenerative medicine.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2024126
Transcription factor 3 (TCF3), a pivotal member of the TCF/LEF family, plays a critical role in tumorigenesis. Nonetheless, its impact on the tumor microenvironment (TME) and cancer phenotypes remains elusive. We perform an exhaustive analysis of TCF3 expression, DNA variation profiles, prognostic implications, and associations with the TME and immunological aspects. This study is based on a large-scale pan-cancer cohort, encompassing over 17,000 cancer patients from multiple independent datasets, validated by in vitro assays. Our results show that TCF3/4/7 exhibits differential expression patterns between normal and tumor tissues across pan-cancer analyses. Mutational analysis of TCF3 across diverse cancer types reveals the highest alteration rates in biliary tract cancer. Additionally, mutations and single nucleotide variants in TCF3/4/7 are found to exert varied effects on patient prognosis. Importantly, TCF3 emerges as a robust predictor of survival across all cancer cohorts and among patients receiving immune checkpoint inhibitors. Elevated TCF3 expression is correlated with more aggressive cancer subtypes, as validated by immunohistochemistry and diverse cohort data. Furthermore, TCF3 expression is positively correlated with intratumoral heterogeneity and angiogenesis. In vitro investigations demonstrate that TCF3 is involved in epithelial-mesenchymal transition, migration, invasion, and angiogenesis. These effects are likely mediated through the interaction of TCF3 with the NF-κB/MMP2 pathway, which is modulated by IL-17A in human uveal melanoma MUM2B cells. This study elucidates, for the first time, the significant associations of TCF3 with DNA variation profiles, prognostic outcomes, and the TME in multiple cancer contexts. TCF3 holds promise as a molecular marker for diagnosis and as a potential target for novel therapeutic strategies, particularly in uveal melanoma.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2024166
EGFR protein trafficking is critical for regulating multiple biological processes, including cell growth and survival. However, how EGFR protein homeostasis is maintained remains unclear. In this study, we show that a reduction in plasma membrane-associated EGFR triggers EGFR transcription by promoting pSTAT3 nuclear localization. Nucleus-localized pSTAT3 binds to the EGFR gene promoter to transactivate EGFR. Moreover, erlotinib, an EGFR tyrosine kinase inhibitor (TKI), can also increase pSTAT3 nuclear accumulation, resulting in increased EGFR transcription and erlotinib resistance. Importantly, pharmacological inhibition of pSTAT3 can significantly overcome the resistance of cancer cells to erlotinib. Together, these findings demonstrate that pSTAT3 is pivotal for maintaining EGFR protein homeostasis and suggest that activation of the pSTAT3-EGFR axis contributes to EGFR-TKI resistance.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2025083
The urea cycle occurs mainly in the liver and undergoes changes during hepatocarcinogenesis. Argininosuccinate synthase 1 (ASS1) is a key enzyme in the urea cycle and is expressed at low levels in certain cancers. LM2I, a specific activator of ASS1, exhibits significant antitumor activity. However, the antitumor mechanism of LM2I in liver cancer remains unclear. In this study, we find that LM2I is more effective for liver cancer cells with low ASS1 expression. The results of the IP-LC/MS experiments reveal that ASS1 interacts with CAD. The expressions of ASS1 and CAD in liver cancer tissues and cells are negatively correlated. LM2I promotes the ubiquitination of CAD protein through ASS1. LM2I inhibits the proliferation of liver cancer cells in vivo and in vitro. However, its efficacy is weak in liver cancer cells stably overexpressing CAD. The H&E staining results reveal that LM2I has no toxicity in mice. In terms of metabolism, LM2I increases the urea content and decreases the pyrimidine content in liver cancer cells. Overexpression of CAD can reduce the inhibitory effect of LM2I on pyrimidine. Pyrimidine supplementation facilitates the proliferation of liver cancer cells, particularly when they are treated with LM2I. In summary, ASS1 interacts with CAD, and LM2I enhances CAD degradation through the activation of ASS1, consequently inhibiting pyrimidine synthesis and the progression of liver cancer.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2025101
Abnormal proliferation and migration of endothelial cells are key contributors to tumor angiogenesis. Recent studies have shown that the crucial role of E3 ubiquitin ligase neuronal precursor cell expression developmentally downregulated 4-like (NEDD4L) in tumorigenesis. However, the precise mechanisms by which NEDD4L functions in endothelial cells remain unclear. In this study, we investigate the mechanisms by which NEDD4L influences the function of human umbilical vein endothelial cells (HUVECs) and its effect on tumor angiogenesis. Our results show that NEDD4L overexpression in HUVECs suppresses both cell proliferation and migration. Additionally, we find that the autophagic activity in NEDD4L-overexpressing cells is increased. Proteomic profiling and ubiquitination assays reveal that NEDD4L interacts with eEF1A1, promoting K48-linked ubiquitination-mediated degradation of eEF1A1. This post-translational modification is a key step in the NEDD4L-mediated regulation of autophagy and cellular function. Moreover, we find that loss of endothelial NEDD4L significantly enhances tumor growth and promotes angiogenesis in vivo. Overall, NEDD4L plays a crucial role in inhibiting tumor angiogenesis by regulating eEF1A1 ubiquitination and degradation, providing new insights into the NEDD4L-eEF1A1 axis and its potential as a therapeutic target.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2024189
Osteoarthritis (OA) is a significant contributor to pain and disability worldwide. Pain is the main complaint of OA patients attending the clinic and has a large impact on their quality of life and economic standards. However, existing treatments for OA-related pain have not been shown to achieve good relief. The main focus is on preventing and slowing the progression of OA so that the problem of OA pain can be resolved. Pain caused by OA is complex, with the nature, location, duration, and intensity of pain changing as the disease progresses. Previous research has highlighted the role of various forms of cell death, such as apoptosis and necrosis, in the progression of pain in OA. Emerging studies have identified additional forms of novel cell death, such as pyroptosis, ferroptosis, and necroptosis that are linked to pain in OA. Different types of cell death contribute to tissue damage in OA by impacting inflammatory responses, reactive oxygen species (ROS) production, and calcium ion levels, ultimately leading to the development of pain. Evidence suggests that targeting novel types of cell death could help alleviate pain in OA patients. This review delves into the complex mechanisms of OA pain, explores the relationship between different modes of novel cell death and pain, and proposes novel cell death as a viable strategy for the treatment of these conditions, with the goal of providing scientific references for the development of future OA pain treatments and drugs.
Acta Biochimica et Biophysica Sinica•2024•DOI: 10.3724/abbs.2024037
This is a corrigendum to the original article. The original article reported that Huperzine A ameliorates neurological deficits after spontaneous subarachnoid hemorrhage through endothelial cell pyroptosis inhibition. The corrigendum corrects errors in the author list and affiliations. The corrected authors and affiliations are provided.
Acta Biochimica et Biophysica Sinica•2024•DOI: 10.3724/abbs.2024140
There are three isoforms of human collagen prolyl 4-hydroxylases (C-P4Hs), each of which has been reported to play an important role in regulating the progression of a variety of human cancers. By analyzing TGCA datasets on human head and neck squamous cell carcinoma (HNSC), we find that a higher expression of all three C-P4HAs (the α subunit of C-P4Hs) is a superior prognostic indicator than a higher expression of two or a single C-P4HA. Unexpectedly, some patients with higher levels of three C-P4HAs survive longer than patients whose tumors have lower expression of C-P4HAs. Therefore, there may be molecule(s) that can negate the deleterious effects of overexpressing C-P4HAs during cancer progression. By constructing a functional protein interaction network of C-P4HAs and analyzing molecules whose expressions are correlated significantly with that of C-P4HAs, we identify scribble cell polarity complex component 2 (LLGL2) as a factor that antagonizes the effects of overexpressed C-P4HAs on HNSC. Silencing of LLGL2 in the human oral squamous cell line Cal-27 upregulates the expression of occludin and increases cancer cell invasion and migration. In contrast, knocking down C-P4HA alone inhibits cell migration and invasion. Furthermore, simultaneously downregulating three C-P4HAs has more pronounced effects on inhibiting cell migration and invasion. Accordingly, high LLGL2 expression is also a marker indicating improved prognosis in patients with HNSC. These results suggest that the interplay between LLGL2 and C-P4HAs may be targeted to mitigate HNSC tumorigenesis and progression.
Acta Biochimica et Biophysica Sinica•2024•DOI: 10.3724/abbs.2024035
Acute lung injury (ALI) is a serious condition characterized by damage to the lungs. Recent research has revealed that activation of the NLRP3 inflammasome in alveolar macrophages, a type of immune cell in the lungs, plays a key role in the development of ALI. This process, known as pyroptosis, contributes significantly to ALI pathogenesis. Researchers have conducted comprehensive bioinformatics analyses and identified 15 key genes associated with alveolar macrophage pyroptosis in ALI. Among these, NLRP3 has emerged as a crucial regulator. This study further reveal that the ULK1 protein diminishes the expression of NLRP3, thereby reducing the immune response of alveolar macrophages and mitigating ALI. Conversely, TRAF3, another protein, is found to inhibit ULK1 through a process called ubiquitination, leading to increased activation of the NLRP3 inflammasome and exacerbation of ALI. This TRAF3-mediated suppression of ULK1 and subsequent activation of NLRP3 are confirmed through various in vitro and in vivo experiments. The presence of abundant M0 and M1 alveolar macrophages in the ALI tissue samples further support these findings. This research highlights the TRAF3-ULK1-NLRP3 regulatory axis as a pivotal pathway in ALI development and suggests that targeting this axis could be an effective therapeutic strategy for ALI treatment.
Acta Biochimica et Biophysica Sinica•2024•DOI: 10.3724/abbs.2024021
Diabetic nephropathy (DN) is a severe complication of diabetes and the leading cause of end-stage renal disease and death. Germacrone (Ger) possesses anti-inflammatory, antioxidant and anti-DN properties. However, it is unclear whether the improvement in kidney damage caused by Ger in DN mice is related to abnormal compositions and metabolites of the gut microbiota. This study generates a mouse model of DN to explore the potent therapeutic ability and mechanism of Ger in renal function by 16S rRNA sequencing and untargeted fecal metabolomics. Although there is no significant change in microbiota diversity, the structure of the gut microbiota in the DN group is quite different. Serratia_marcescens and Lactobacillus_iners are elevated in the model group but significantly decreased after Ger intervention (P<0.05). Under the treatment of Ger, no significant differences in the diversity and richness of the gut microbiota are observed. An imbalance in the intestinal flora leads to the dysregulation of metabolites, and non-targeted metabolomics data indicate high expression of stearic acid in the DN group, and oleic acid could serve as a potential marker of the therapeutic role of Ger in the DN model. Overall, Ger improves kidney injury in diabetic mice, in part potentially by reducing the abundance of Serratia_marcescens and Lactobacillus_iners, as well as regulating the associated increase in metabolites such as oleic acid, lithocholic acid and the decrease in stearic acid. Our research expands the understanding of the relationship between the gut microbiota and metabolites in Ger-treated DN. This contributes to the usage of natural products as a therapeutic approach for the treatment of DN via microbiota regulation.
Acta Biochimica et Biophysica Sinica•2024•DOI: 10.3724/abbs.2024121
Gene regulation refers to the precise regulation of gene expression in an organism, and transcription factors are proteins that bind to DNA and regulate gene expression by promoting or inhibiting the expressions of target genes. Since the late 1980s [1], scientists have studied special genes called Pax genes that control how genes function in organisms as they grow. There are nine Pax genes found in animals such as mice, zebrafish, and humans [2]. Based on the composition domain and homology of the sequence, the Pax family is divided into four subfamilies: Pax1/9, Pax2/5/8, Pax4/6, and Pax3/7 [3]. Pax7 plays a pivotal role in the implementation, protection, and repair of skeletal muscle. Pax7 helps to control the balance between self-renewal and differentiation of satellite cells, ensuring that they can proliferate when needed to generate new muscle cells and differentiate into mature muscle fibers when necessary for muscle development and repair. The expression of the Pax7 gene in nerve cells is critical for dorsal root and sensory ganglia development. The Pax7 gene serves as a primary controlling factor for skeletal muscle development while influencing different biological processes; however, its exact role in jawless vertebrates such as lamprey remains unclear, and extensive research is needed to elucidate the intricate underlying mechanisms involved. Given the unique status of lamprey as an ancient jawless fish, possessing an ancient lineage and distinctive biological features, it is rare to explore gene function across hundreds of millions of years of vertebrate evolution. The use of lamprey as a model system for gene function research represents an innovative approach in the fields of evolutionary and comparative genomics. In this study, we investigated the regulatory mechanism of Pax7 in lamprey via gene cloning, gene expression analysis, gene silencing and transcriptome data analysis. We also explored the interactions between genes with significant differences. Identification of Lr.Pax7 in lamprey tissues began with the retrieval of protein sequences that are similar to those of human Pax family members in sea lamprey (Petromyzon marinus) or zebrafish (Danio rerio) from the NCBI protein database (Supplementary Table S1) and the use of BLAST to identify corresponding homologs (Supplementary Table S2). Subsequently, we extracted the Pax sequences from our library. Lethenteron reissneri specimens were dissected to isolate various tissues. Primers targeting the pax domains were designed based on the Pax7 nucleotide sequence in the Lampreys cDNA library, and the aim was to verify the effectiveness of the Lampreys cDNA as a template for validation (Supplementary Table S3). Lr.Pax7 was successfully amplified via PCR in muscle tissue. Here, a variety of methods were used for bioinformatics analysis. The results showed that the amino acid sequence of Pax7 is highly similar among animals (Figure 1A), with a decreasing trend from higher to lower organisms, as revealed by sequence alignment. It can be observed from the evolutionary tree (Figure 1B) that Pax genes for each subfamily are present in ancestral chordate and that Pax genes are present in amphioxus. Petromyzon marinus, Lethenteron camtschaticum, and Lethenteron reissneri constitute a sister group and have become good models for the study of jawless vertebrates. Pax9, Pax2, Pax6, and Pax7 show high similarity to those of other higher vertebrates. Therefore, these genes were named Lr.Pax9, Lr.Pax2, Lr.Pax7, and Lr.Pax6. The results indicate that the Pax7 gene is significantly preserved across various species, from higher to lower. This suggests that the DNA sequence of the gene is remarkably similar among different species. Lr.Pax7 is positioned between vertebrates and invertebrates and is most closely related to P. marinus Pax7. This finding provides more insight into the original evolutionary position of the lamprey. Crystal structure prediction analysis revealed that Lr.Paxs and Hm.Paxs have highly homologous structures (Figure 1C). The Pax gene has a similar structure (Figure 1D), including a conserved DNA-binding structure called the pair-box domain. This structure contains approximately 128 amino acids and is responsible for binding with specific DNA sequences, regulating gene expression, and interacting with other proteins. To further investigate the evolutionary history of Pax7 in vertebrates, we compared the genetic environment of Pax7 with that of other vertebrates (Figure 1E). In addition, many Pax gene members also contain DNA-binding structures called homeodomains, which play important roles in development.
Acta Biochimica et Biophysica Sinica•2024•DOI: 10.3724/abbs.2024089
Growth hormone deficiency (GHD) is the most common pituitary hormone deficiency and is clinically characterized by short stature, delayed bone age and central distribution of body fat, and it has also been proven to be mildly heritable. Treatment with recombinant human growth hormone (r-hGH) is primary and safe for GHD children, and a dose of 0.15‒0.20 mg/kg each week results in a considerable increase in height velocity, with noteworthy growth during the first year of therapy [1]. Previous studies have shown that serum IGF-1 is strongly correlated with the growth response [2]. Therefore, IGF-1 can serve as a clinical indicator for monitoring compliance, efficacy and safety. However, the response to GH therapy shows significant individual variation, which is strongly associated with genetic factors. The prevalence rate of severe childhood GHD-related short stature varies from 1:4000 to 1:10,000 [3], while approximately 3%‒4% of the population in China suffers from short stature with an increasing trend. Therefore, an open-label, prospective, multicentric, noncomparative, nonrandomized phase IV interventional study (NCT01187550, Merck Serono Study 27709) was conducted to investigate the relationship between the prospective biomarkers of GHD patients and the individual variation in the primary therapeutic response following 4 weeks of r-hGH therapy. Given the significance of predicting GHD treatment response and the gaps in previous research, we sought to adopt a comprehensive strategy to accurately predict the therapeutic response utilizing the transcriptome, single nucleotide polymorphisms (SNPs) and clinical factors. We employed continuous variables and standard deviation scores of differences in serum IGF-1 levels after 4 weeks of r-hGH therapy (ΔIGF-1) as targets to filter possible influencing variables. Furthermore, we compared several potential machine learning techniques, validated by PCA and PLS-DA, and ultimately applied the elastic net algorithm to determine the optimized predictive factors with consistent effect sizes. Additionally, expression quantitative trait locus (eQTL) analysis and differentially expressed gene (DEG) analysis were conducted to identify significant biomarkers for GHD treatment.
Acta Biochimica et Biophysica Sinica•2024•DOI: 10.3724/abbs.2024076
Rheumatoid arthritis (RA) is an idiopathic and chronic autoimmune disease for which there are currently no effective treatments. Oxypeucedanin hydrate (OXH) is a natural coumarin known for its potent anti-inflammatory properties. However, further investigations are needed to determine its therapeutic efficacy in treating RA. In this study, we evaluate the anti-inflammatory activity of OXH by treating LPS-induced RAW264.7 macrophages. Our results show that OXH treatment reverses the changes in iNOS, COX-2, IL-1β, IL-6, and TNF-α levels. Additionally, OXH reduces ROS production. Further analysis reveals that OXH suppresses the activation of the NF-κB/MAPK pathway. CETSA results show that OXH competes with LPS for binding to the TLR4/MD2 complex. MST experiments demonstrate the specific affinity of OXH for the TLR4/MD2 complex, with a Kd value of 33.7 μM. Molecular docking analysis suggests that OXH binds to the pocket of the TLR4/MD2 complex and interacts with specific amino acids, such as GLY-343, LYS-388, and PHE-345. Molecular dynamics simulations further confirm this conclusion. Finally, we investigate the potential of OXH in treating RA using a collagen-induced arthritis (CIA) model in rats. OXH effectively ameliorates the symptoms of CIA, including improving body weight, reducing swelling and redness, increasing talus volume, and decreasing bone erosion. OXH also decreases the mRNA levels of pro-inflammatory factors in synovial tissue. Transcriptome enrichment analysis and western blot analysis confirm that OXH suppresses the NF-κB/MAPK pathway, which is consistent with our in vitro findings.
Acta Biochimica et Biophysica Sinica•2024•DOI: 10.3724/abbs.2024039
Vaccines play essential roles in the fight against the COVID-19 pandemic. The development and assessment of COVID-19 vaccines have generally focused on the induction and boosting of neutralizing antibodies targeting the SARS-CoV-2 spike (S) protein. Due to rapid and continuous variation in the S protein, such vaccines need to be regularly updated to match newly emerged dominant variants. T-cell vaccines that target MHC I- or II-restricted epitopes in both structural and non-structural viral proteins have the potential to induce broadly cross-protective and long-lasting responses. In this work, the entire proteome encoded by SARS-CoV-2 (Wuhan-hu-1) is subjected to immunoinformatics-based prediction of HLA-A*02:01-restricted epitopes. The immunogenicity of the predicted epitopes is evaluated using peripheral blood mononuclear cells from convalescent Wuhan-hu-1-infected patients. Furthermore, predicted epitopes that are conserved across major SARS-CoV-2 lineages and variants are used to construct DNA vaccines expressing multi-epitope polypeptides. Most importantly, two DNA vaccine constructs induce epitope-specific CD8+ T-cell responses in a mouse model of HLA-A*02:01 restriction and protect immunized mice from challenge with Wuhan-hu-1 virus after hACE2 transduction. These data provide candidate T-cell epitopes useful for the development of T-cell vaccines against SARS-CoV-2 and demonstrate a strategy for quick T-cell vaccine candidate development applicable to other emerging pathogens.
Acta Biochimica et Biophysica Sinica•2024•DOI: 10.3724/abbs.2023287
Lung adenocarcinoma (LUAD) is one of the most aggressive types of lung cancer. The prognosis of LUAD patients remains poor, and the overall efficacy of gemcitabine-based chemotherapy is still unsatisfactory. Long noncoding RNAs (lncRNAs) play important roles in several cancer types by interacting with multiple proteins, RNA, and DNA. However, the relationship between lncRNA dysregulation and gemcitabine resistance in LUAD has not been fully elucidated. In this study, lncRNA CYTOR expression and its association with the prognosis of LUAD patients are assessed by quantitative RT-PCR and Kaplan-Meier survival analysis. In vitro and in vivo functional studies are conducted to evaluate the biological functions of CYTOR in LUAD. The underlying mechanism regarding the tumor-promoting effects of CYTOR is explored using RNA immunoprecipitation, biotin-labelled RNA pulldown, luciferase reporter assays, and western blot analysis. We identify that CYTOR is an oncogenic lncRNA and is apparently upregulated in LUAD by analysing TCGA-LUAD data. High CYTOR expression is a poor prognostic factor for LUAD. Functional studies reveal that CYTOR confers LUAD cells with stronger resistance to gemcitabine treatment and upregulates the expression levels of epithelial-mesenchymal transition (EMT)-related proteins. Mechanically, CYTOR acts as a competitive endogenous RNA (ceRNA) to absorb miR-125a-5p, weakens the antitumor function of miR-125a-5p, and ultimately upregulates ANLN and RRM2 expressions. Taken together, this study explains the mechanism of lncRNA in the gemcitabine resistance of LUAD and formulates a theoretical framework for the in depth study of LUAD.
Chinese Journal of Pathophysiology•2025•DOI: 10.3969/j.issn.1000-4718.2025.09.002
AIM: To investigate whether melatonin can ameliorate acute myocardial infarction (AMI) by inhibiting ferroptosis. METHODS: H9C2 cells were cultured in AnaeroPack system with low sugar and serum-free medium for 10 h to construct a cell model of AMI. Then cells were treated with melatonin and ferroptosis inducer erastin. The cell activity, reactive oxygen species (ROS), lipid peroxidation, mitochondrial membrane potential (MMP), and ferroptosis related protein expression were detected. A rat model of AMI induced by isoprenaline (ISO) injection was established to evaluate the effects of melatonin, in which the myocardial infarction size, cardiac injury, pathological changes, oxidative stress, iron ion and ferroptosis related protein expression were examined. RESULTS: Melatonin decreased the oxidative stress, lipid peroxidation and expression of ferroptosis protein in cardiomyocytes induced by hypoxia, but these effects could be impeded by the ferroptosis inducer erastin. Furthermore, in vivo experiments, we also found that melatonin improved the myocardial infarction size, cardiac injury, pathological changes, oxidative stress, and alleviated iron ion accumulation and ferroptosis. CONCLUSION: The cardioprotective effects of melatonin in AMI are associated with the inhibition of ferroptosis.
Chinese Journal of Pathophysiology•2025•DOI: 10.3969/j.issn.1000-4718.2025.12.001
AIM: To evaluate the function and mechanisms of forskolin in treating ataxia-like behavior in Celsr3 conditional knockout (cKO) mice. METHODS: The efficiency of intraperitoneally administered forskolin was evaluated by behavioral tests, and the molecular mechanisms were investigated by patch-clamp experiments. RESULTS: The loss of Celsr3 led to ataxia-like behavior, accompanied by impaired miniature excitatory postsynaptic currents (mEPSCs) and postsynaptic long-term potentiation (LTP) in PCs. The cAMP activator forskolin ameliorated ataxia-like behavior and abrogated the mEPSCs impairment and LTP in model mice. Interestingly, the effects of forskolin could be blocked by SQ22536 (a cAMP antagonist) and ESI-08 (exchange protein activated by cAMP antagonist; Epac) but the H89 (a PKA antagonist) could not block the effects. CONCLUSION: Celsr3 plays an important role in motor coordination by modulating synaptic function, and forskolin may be a valuable therapeutic drug for certain types of inherited cerebellar ataxia.
Stem Cell Research & Therapy•2026•DOI: 10.1186/s13287-026-05051-z
Intervertebral disc degeneration (IVDD) is a leading cause of low back pain, driven by mitochondrial dysfunction and impaired mitophagy in nucleus pulposus-derived mesenchymal stem cells (NPMSCs). Sirtuin 1 (SIRT1), a key regulator of mitochondrial quality control, is downregulated in degenerated discs. Salidroside (Sal), a natural compound from Rhodiola rosea, has shown potential in enhancing mitophagy, but its mechanism in IVDD remains unclear. Using network pharmacology, molecular docking, and dynamics simulations, we identified SIRT1 as a key target of Sal. Human and rat NPMSCs were isolated and treated with tert-butyl hydroperoxide (TBHP) to induce degeneration. In vitro assays included CCK-8, EdU, SA-β-Gal, JC-1, Western blot, immunofluorescence, and transmission electron microscope (TEM). An in vivo rat IVDD model was established via needle puncture and treated with Sal and/or the autophagy inhibitor 3-MA. Histological, immunohistochemical, and imaging analyses were performed to evaluate IVDD. Sal bound stably to SIRT1 and activated SIRT1/FOXO3 pathway, promoting mitophagic flux, reducing reactive oxygen species accumulation, and suppressing apoptosis in NPMSCs. SIRT1 knockdown or 3-MA treatment abolished Sal’s protective effects. In vivo, Sal treatment preserved disc height, reduced apoptosis, and enhanced mitophagy, while 3-MA exacerbated degeneration. These effects are closely associated with the activation of SIRT1/FOXO3 signaling and the restoration of functional mitophagic flux. Our work highlights the SIRT1/FOXO3-mitophagy axis as a promising target for further investigation in the development of therapeutic strategies for IVDD.
Stem Cell Research & Therapy•2026•DOI: 10.1186/s13287-026-05073-7
Exosomes have emerged as critical mediators of intercellular and inter-organ communication in bone biology. Secreted by bone-resident cells such as osteoblasts, osteoclasts, osteocytes, and mesenchymal stem cells (MSCs), these nanosized vesicles carry diverse molecular cargos that regulate bone remodeling, regeneration, and skeletal homeostasis. In addition to mediating local communication within the bone microenvironment, exosomes also participate in systemic crosstalk between bone and other tissues, including skeletal muscle, adipose tissue, gut microbiota, the immune system, the nervous system, and vasculature. Disruption of these exosome-mediated pathways contributes to the development and progression of bone diseases, including osteoporosis, osteoarthritis, osteonecrosis of the femoral head, and bone metastases. This review summarizes current advances in exosome-mediated signaling in both physiological and pathological contexts, with particular emphasis on their roles as biomarkers, therapeutic agents, and drug delivery vehicles. We also discuss the emerging contribution of artificial intelligence (AI) to exosome research, especially in biomarker discovery, disease classification, and target identification, as well as the major challenges that currently limit clinical translation. Together, these insights highlight the potential of exosome-based strategies for precision medicine in bone diseases.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2025222
Poultry production faces escalating challenges from intensive farming practices, where stressors disrupt intestinal integrity, microbiota balance, and antioxidant defenses, leading to economic losses. Although antibiotics have historically mitigated such issues, growing restrictions due to antimicrobial resistance necessitate natural alternatives. Fungal polysaccharides (FP), notably lentinan (LNT) from Lentinula edodes and polysaccharide from Ganoderma lucidum (GLP), are promising candidates owing to their immunomodulatory, antioxidant, and prebiotic properties. However, existing research focuses predominantly on individual FP, neglecting potential synergies in blended formulations. We hypothesized that mixed FP synergistically enhance intestinal health by simultaneously improving nutrient absorption, activating antioxidant pathways, and stabilizing microbial ecosystems. To investigate, 240 one-day-old Arbor Acres male broilers were randomly assigned to control (0 mg/kg FP) and treatment groups receiving 200, 400, or 600 mg/kg mixed FP (1:1 ratio of GLP and LNT). On day 42, intestinal segments were collected for morphological analysis, antioxidant enzyme activities, gene expression, and cecal microbiota composition. Results showed that 400 mg/kg FP significantly increased villus height and VH/CD ratio across all intestinal segments while reducing crypt depth, indicating enhanced nutrient absorption. Antioxidant enzyme activities (T-AOC, T-SOD, GSH-Px) were elevated, and mRNA expression of HO-1, NQO1, CAT, and Nrf2 was upregulated, with Keap1 downregulated, suggesting activation of the Keap1-Nrf2 pathway. Microbiota analysis revealed increased alpha diversity (Shannon and Simpson indices) and altered composition, with elevated abundances of beneficial genera such as Butyricimonas and Alistipes, and increased Verrucomicrobiota phylum. These findings demonstrate that mixed FP supplementation at 400 mg/kg improves intestinal health, antioxidant capacity, and microbiota diversity in broilers, offering a natural alternative to antibiotics.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2025253
Kidney renal clear cell carcinoma (KIRC) is the most common and aggressive subtype of renal cell carcinoma and is characterized by poor prognosis and high molecular heterogeneity. Long noncoding RNAs (lncRNAs) have emerged as crucial regulators in cancer, yet the functional role of SNHG11 in KIRC remains unclear. In this study, we perform integrated multiomics analysis using data from the TCGA and ICGC cohorts and reveal that SNHG11, a methylation-associated lncRNA, is significantly correlated with poor clinical outcomes. In vitro and in vivo assays demonstrate that SNHG11 promotes tumor proliferation and progression. Mechanistically, SNHG11 enhances oxidative phosphorylation, as evidenced by increased ATP production, disrupted mitochondrial membrane potential, and altered NAD+/NADH ratios. Furthermore, SNHG11 expression is associated with somatic mutation patterns, particularly those involving BAP1 and PBRM1, indicating potential crosstalk between epigenetic regulation and genetic alterations. These findings reveal that SNHG11 is a novel biomarker in KIRC and a potential therapeutic target.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2026037
Breast cancer (BC) is the most common malignancy among women, with approximately 2.3 million new cases diagnosed annually. Brain metastasis is a significant cause of mortality, particularly in HER2-positive and triple-negative subtypes. Current therapies are limited by the blood-brain barrier. This study aimed to identify core genes associated with breast cancer brain metastasis (BCBM) using bioinformatics and machine learning. We analyzed the GSE43837 dataset (19 nonmetastatic primary breast tumors and 19 brain metastases) using differential expression analysis and weighted gene coexpression network analysis (WGCNA). We identified 245 upregulated and 188 downregulated genes. WGCNA revealed key modules (midnightblue and black) with 89 candidate genes. Intersection with differentially expressed genes yielded 29 overlapping genes. LASSO regression and random forest identified four core genes: B3GNT9, SERPINF1, LUM, and CILP. ROC analysis showed strong discriminatory power (AUC > 0.87). External validation in GSE125989 confirmed downregulation of SERPINF1, LUM, and CILP in brain metastases, with a combined model achieving AUC 0.984. Experimental validation in zebrafish and mouse models confirmed the role of these genes in BCBM. These findings suggest that SERPINF1, LUM, CILP, and B3GNT9 are potential biomarkers and therapeutic targets for BCBM.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2026010
Persicaria chinensis, a well-known traditional Chinese medicinal herb that is both edible and medicinal, has been widely acknowledged for its therapeutic effects, such as anti-inflammatory, antioxidant, and antitumor activities. However, the role of miRNAs from this plant in the cross-kingdom regulation of human diseases has not been investigated. In this study, we analyze the miRNA expression profile of P. chinensis using high-throughput sequencing and identify a total of 673 miRNAs, including 422 novel miRNAs that are unique to this plant and 251 conserved miRNAs. Among the conserved miRNAs, pch-miR319a is found to be the most abundant. Moreover, food-oriented pch-miR319a accumulates in the uterus and tumors and exhibits a rich repertoire of target genes within cancer-related pathways, demonstrating significant cross-kingdom regulatory potential. Utilizing the dual-luciferase reporter gene assay, we demonstrate that pch-miR319a from P. chinensis targets the Itga3 gene, which is associated with cervical cancer progression. Overexpression of pch-miR319a significantly decreases the viability, migration, and induces apoptosis of HeLa cervical cancer cells in vitro. Moreover, in a syngeneic mouse tumor model of cervical cancer, treatment with pch-miR319a effectively inhibits tumor growth and downregulates the expressions of ITGA3 and the proliferation marker Ki-67. Our study highlights the potential of pch-miR319a from P. chinensis as a novel therapeutic agent for cervical cancer by targeting ITGA3 and provides new insights into the cross-kingdom regulatory mechanisms of plant miRNAs in human diseases.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2025237
Exposure to chronic hypobaric hypoxia provokes marked alterations in the gut microbiota and its metabolome, yet the functional significance of histidine-derived metabolites in hypobaric hypoxic pulmonary hypertension (PH) remains underexplored. Here, we employ 16S rDNA, metagenomic, and untargeted metabolomic sequencing to characterize longitudinal shifts in the fecal microbiota and metabolites during hypobaric hypoxic PH development in Sprague-Dawley rats. Fecal carnosine levels and the abundance of its producer, Ruminococcus bromii, both decrease significantly over 28 days of hypobaric hypoxia (P < 0.05). Spearman correlation shows that carnosine is inversely correlated with the percentage of pulmonary arteriole media thickness (MT%; r = –0.8741, P < 0.001). Therapeutic supplementation with carnosine restores systemic and pulmonary antioxidant defenses and attenuates vascular remodeling without altering right ventricular pressures. In vitro, carnosine inhibits hypoxia-induced pulmonary artery smooth muscle cell (PASMC) proliferation and migration and suppresses nuclear factor erythroid 2-related factor 2 (Nrf2) accumulation. These findings reveal dynamic gut-lung crosstalk in hypobaric hypoxic PH and nominate carnosine as a metabolite-based intervention to mitigate hypoxia-driven pulmonary vascular remodeling.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2026027
Phosphorylated CTD Interacting Factor 1 (PCIF1) is the exclusive methyltransferase responsible for the N6,2-O-dimethyladenosine (m6Am) modification in mammalian mRNA. Our previous research identified PCIF1 as a potent tumor suppressor in glioma, demonstrating its ability to impair cell proliferation, induce G2/M phase arrest, and promote apoptosis. However, its role in glioma cell migration and invasion remains unclear. In this study, we investigate how PCIF1 regulates glioma cell migration and invasion. Overexpression of PCIF1 inhibits migration and invasion, whereas PCIF1 knockdown enhances these behaviors. Corresponding changes are observed in mesenchymal markers (Vimentin, β-catenin, Snail, Slug) and the epithelial marker T-cadherin, indicating that PCIF1 suppresses epithelial-to-mesenchymal transition (EMT)-mediated glioma invasion. Mechanistically, PCIF1 modulates the AKT pathway by promoting proteasomal degradation of AKT while increasing phosphorylated AKT (p-AKT) levels, revealing a complex regulatory mechanism. PCIF1 knockdown upregulates INPP5B, a lipid phosphatase, causing accumulation of PI(3,4)P2 and enhanced AKT activation. Conversely, PCIF1 overexpression increases PI(3,4,5)P3 production, elevating p-AKT levels. This bidirectional regulation suggests that PCIF1 influences phosphoinositide signaling and AKT activation. Our findings highlight PCIF1 as a key modulator of glioblastoma cell migration and invasion through phosphoinositide signaling, positioning it as a potential biomarker and therapeutic target in glioma.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21235
BACKGROUND: Factors such as infection, limb ischemia, and histiocyte activation are involved in diabetic foot ulcers, but the key cell subpopulations influencing diabetic foot ulcer healing remain unclear, and specific biomarkers for diabetic foot ulcers have yet to be identified. Gene Expression Omnibus (GEO) is a publicly accessible database managed by the National Center for Biotechnology Information that stores high-throughput gene expression data, allowing users to freely submit, share, query, and analyze data. Secondary analysis of published data can save research costs and uncover new research targets and ideas. OBJECTIVE: To screen biomarkers for diabetic foot ulcers using single-cell transcriptome and conventional transcriptome bioinformatics analysis, high-dimensional weighted gene co-expression network analysis (hdWGCNA), and weighted gene co-expression network analysis (WGCNA). METHODS: The single-cell transcriptome dataset GSE165816, containing non-healing ulcer tissue samples from diabetic foot ulcer patients and foot skin samples from diabetic patients, was downloaded from GEO. After data quality control, dimensionality reduction, differential analysis, cell type annotation, and pseudotime analysis, cell types spanning the entire course of diabetic foot ulcers were identified, and differentially expressed genes (DEGs) were obtained. hdWGCNA identified gene modules highly correlated with diabetic foot ulcers. Conventional transcriptome datasets GSE68183 and GSE80178, containing non-healing ulcer tissue samples from diabetic foot ulcer patients and foot skin samples from diabetic patients, were downloaded for differential analysis to screen DEGs, and WGCNA was used to identify diabetic foot ulcer-related gene modules. The DEGs from single-cell transcriptome, DEGs from conventional transcriptome samples, and module genes from WGCNA and hdWGCNA were integrated to screen biomarkers for diabetic foot ulcers. The GSE134431 dataset was downloaded as a validation conventional transcriptome dataset, and the expression levels of diabetic foot ulcer biomarkers were compared in single-cell transcriptome and validation conventional transcriptome datasets. Diabetic and diabetic foot ulcer rat models were replicated, wound tissue was collected, and immunohistochemistry and western blot were used to detect biomarker expression levels. RESULTS AND CONCLUSION: Single-cell transcriptome data analysis showed that epithelial cell differentiation spanned the entire pathological process of diabetic foot ulcers. A total of 146 DEGs were obtained from single-cell transcriptome between groups, including 59 upregulated and 87 downregulated DEGs. hdWGCNA identified 19 gene modules related to diabetic foot ulcers, containing 476 core genes. Conventional transcriptome data analysis yielded a total of 913 DEGs, including 343 upregulated and 570 downregulated DEGs. WGCNA obtained 19 diabetic foot ulcer-related gene modules, containing 887 genes. Two biomarkers for diabetic foot ulcers were screened: S100A14 and SFN. The expression levels of these two genes in diabetic foot ulcer samples were higher than those in diabetic foot skin samples in both single-cell transcriptome and validation conventional transcriptome datasets. Animal experiments showed that the expression levels of S100A14 and SFN in wound tissue of diabetic foot ulcer rats were higher than those in back skin tissue of diabetic rats. The results indicate that the pathological process of diabetic foot ulcers involves multiple cell types, among which epithelial cells are the key cell subpopulation. S100A14 and SFN are significantly upregulated in diabetic foot ulcer samples and are potential targets for the treatment of diabetic foot ulcers.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21279
Objective: To systematically compare the acute effects of blood flow restriction combined with preconditioning (to induce post-activation performance enhancement) versus preconditioning alone or sitting, low-intensity preconditioning combined with blood flow restriction versus high-intensity preconditioning, and sitting combined with blood flow restriction versus sitting on sports performance using a multilevel meta-analysis. Methods: Following the PRISMA guidelines, Web of Science, PubMed, SPORTDiscus, and CNKI databases were systematically searched (from inception to May 24, 2025). Inclusion criteria: (1) healthy individuals who were at least physically active; (2) studies with at least one of the following four comparisons: preconditioning + blood flow restriction vs. preconditioning alone; preconditioning + blood flow restriction vs. sitting; low-intensity preconditioning + blood flow restriction vs. high-intensity preconditioning; sitting + blood flow restriction vs. sitting; (3) sports performance (e.g., jump, sprint, bench press throw) as the primary outcome; (4) randomized or non-randomized crossover/parallel designs; (5) published in peer-reviewed Chinese or English journals. Risk of bias was assessed using ROB-2, and evidence quality was evaluated with GRADE. Data were fitted using cluster robust variance estimation and a three-level mixed-effects model, with small-sample corrections. Subgroup analyses and meta-regression explored moderators and sources of heterogeneity. Results: Twelve studies (196 participants, 12 women, 184 men) were included. Main findings: (1) Preconditioning + blood flow restriction was more effective than preconditioning alone in enhancing sports performance (ES=0.21, 95%CI=0.01-0.40, GRADE=low), with the best effect at recovery times of 4-12 min and 50% arterial occlusion pressure (ES=1.49); (2) Preconditioning + blood flow restriction did not significantly differ from sitting (ES=0.52, 95%CI=-0.12-1.15, GRADE=very low), but preconditioning + 140 mmHg blood flow restriction was superior to preconditioning alone (ES=1.21, 95%CI=0.14-2.28); (3) Low-intensity preconditioning + blood flow restriction did not differ from high-intensity preconditioning (ES=-0.10, 95%CI=-0.84-0.64, GRADE=low); (4) Sitting + blood flow restriction did not significantly differ from sitting (ES=0.24, 95%CI=-0.03-0.52, GRADE=very low). Notably, the effects of the latter two comparisons significantly decreased with recovery time (β=-0.04, P < 0.01 and β=-0.04, P=0.02). Conclusion: Preconditioning combined with blood flow restriction is more effective than preconditioning alone in inducing post-activation performance enhancement, preliminarily suggesting the use of 50% arterial occlusion pressure and 4-12 min recovery time. However, preconditioning combined with blood flow restriction does not appear to be more effective than sitting, possibly due to insufficient number of included studies. Additionally, low-intensity preconditioning + blood flow restriction can achieve similar post-activation performance enhancement as high-intensity preconditioning, while the potential benefit of sitting + blood flow restriction on sports performance may diminish over time. Overall, it is preliminarily recommended to use low-intensity preconditioning (e.g., 30% one-repetition maximum squat or bodyweight training) combined with 50% arterial occlusion pressure or 140 mmHg blood flow restriction, with 4-12 min recovery before subsequent performance testing.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21240
BACKGROUND: Exercise preconditioning produces early and late myocardial protective effects, in which protein kinase C and myocardial ATP-sensitive potassium channels (SarcKATP) are mediators and effectors, respectively. Protein kinase C regulates the expression of myocardial SarcKATP channels. OBJECTIVE: To compare the effects of protein kinase C on the expression of myocardial SarcKATP channel subunits, inward recirculating potassium channel 6.2 (Kir6.2) and sulfonylurea receptor 2A (SUR2A), in exercise preconditioning. METHODS: Forty-eight Sprague-Dawley rats were randomly divided into five groups: control group (no intervention), early exercise preconditioning group, protein kinase C inhibitor (pre-exercise intraperitoneal injection) + early exercise preconditioning group, late exercise preconditioning group, and protein kinase C inhibitor + late exercise preconditioning group. After preconditioning, the distribution and expression changes of Kir6.2 and SUR2A mRNAs in the rat myocardium were observed and detected using real-time fluorescent quantitative PCR. The distribution and expression changes of Kir6.2 and SUR2A proteins were observed and detected using western blot. RESULTS AND CONCLUSION: (1) Compared with the control group, the mRNA expression of Kir6.2 and SUR2A showed no significant difference in the early and late exercise preconditioning group. (2) Compared with the early exercise preconditioning group, the protein kinase C inhibitor + early exercise preconditioning group showed decreased Kir6.2 mRNA expression but increased Kir6.2 protein expression; both SUR2A mRNA and protein expression decreased. (3) Compared with the late exercise preconditioning group, the protein kinase C inhibitor + late exercise preconditioning group showed decreased Kir6.2 mRNA and SUR2A mRNA expression, decreased Kir6.2 protein expression, but increased SUR2A protein expression. (4) These results indicate that for the same subunit (Kir6.2 or SUR2A), protein kinase C exerts coordinated and complementary regulatory effects in early and late exercise preconditioning; for different subunits (Kir6.2 and SUR2A), protein kinase C also exerts coordinated and complementary regulatory effects on their expression in early and late exercise preconditioning.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21225
BACKGROUND: The crescent sign is a significant radiological feature in the progression of steroid-induced osteonecrosis of the femoral head (SIONFH), indicating the separation and defect of articular cartilage and subchondral bone. The appearance of the crescent sign is associated with the mid-to-late stages of the disease and poor prognosis. However, studies on the specific pathological characteristics and progression patterns of articular cartilage in SIONFH remain unclear. OBJECTIVE: To observe the pathological features of articular cartilage in specimens from different stages of SIONFH, explore the progression and pathological mechanisms, and elucidate the formation mechanism of the crescent sign, providing a theoretical basis for optimizing hip-preserving strategies. METHODS: Femoral head specimens were collected from patients with SIONFH who underwent total hip arthroplasty at the First Affiliated Hospital of Guangzhou University of Chinese Medicine from 2021 to 2024. According to the ARCO staging, they were divided into mild, moderate, and severe collapse groups, with fresh femoral neck fracture specimens as controls. All specimens were cut coronally, and the folded cartilage surface in the necrotic area was taken; control group took corresponding area. Hematoxylin-eosin staining and Safranin O-fast green staining were used for morphological observation, immunohistochemistry and western blot for biomarker expression, and apoptosis kit for apoptosis level. RESULTS AND CONCLUSION: (1) Gross observation: The control group showed smooth cartilage surface without folds or hyperplasia, no separation or defect between articular cartilage and subchondral bone, and tough texture. In SIONFH specimens, obvious folds were visible on the cartilage surface, with separation and defects between articular cartilage and subchondral bone, and a loose sensation on pressing. (2) Pathological observation: In the control group, chondrocytes in each layer were arranged neatly, cartilage matrix stained uniformly, tidemark was intact and continuous, calcified cartilage layer and subchondral bone connection was clear and complete, and bone trabeculae were arranged neatly. In SIONFH specimens, chondrocytes were disorganized, empty lacunae increased, matrix staining loss of varying degrees, tidemark duplication and loss, calcified cartilage layer showed numerous cavities and sclerosis, with granulation tissue invasion into cavities, separation and defects between calcified cartilage and subchondral bone, and abundant proliferative granulation tissue in subchondral bone trabecular spaces. (3) Immunohistochemistry: In SIONFH specimens, positive staining of Runt-related transcription factor 2, matrix metalloproteinase 13, matrix metalloproteinase 3, and collagen type I alpha 2 chain increased in calcified cartilage layer and deep cartilage; vascular endothelial growth factor A, hypoxia-inducible factor 1 alpha, interleukin-1 beta, and tumor necrosis factor alpha positive staining increased in subchondral bone trabecular spaces and deep cartilage granulation and scar tissue. (4) Western blot results showed decreased expression of collagen type II alpha 1 chain and SOX9, and increased expression of Runt-related transcription factor 2, matrix metalloproteinase 13, hypoxia-inducible factor 1 alpha, and vascular endothelial growth factor A in SIONFH specimens. (5) Caspase3/7 activity in SIONFH samples was significantly higher than that in the control group, positively correlated with the degree of collapse. (6) These results indicate that articular cartilage lesions in SIONFH mainly concentrate in the deep cartilage and calcified cartilage around the necrotic area. Necrosis of subchondral bone leads to changes in local microenvironment and elastic modulus, causing sclerosis of calcified cartilage. With continued weight-bearing, stress concentration at the necrosis-sclerosis junction leads to brittle fracture, which is the starting point of fracture. Bone and cartilage fracture leads to destruction of the subchondral cortical bone barrier, and invasion of granulation tissue from subchondral bone trabecular spaces directly stimulates calcified cartilage and deep cartilage, resulting in terminal differentiation, apoptosis, matrix degradation, and cavity formation of chondrocytes, leading to decreased repair capacity of articular cartilage. The diseased cartilage cannot properly interlock with subchondral bone, and with disease progression, extensive separation and defects eventually appear between bone and cartilage, manifesting as the crescent sign on imaging.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21237
BACKGROUND: Ferroptosis is an important pathological mechanism in spinal cord ischemia-reperfusion injury. Although studies have confirmed that prostaglandin E1 attenuates cerebral microvascular endothelial cell injury in the hippocampus induced by chronic cerebral hypoperfusion, its effect on ferroptosis of endothelial cells after spinal cord ischemia-reperfusion injury remains poorly studied. OBJECTIVE: To investigate whether prostaglandin E1 pretreatment attenuates spinal cord ischemia-reperfusion injury by inhibiting ferroptosis in endothelial cells and to elucidate possible mechanisms. METHODS: (1) Cell experiment: Rat spinal cord microvascular endothelial cells were divided into four groups. Control group was cultured under normoxia (20% O2) with complete medium. Model group was subjected to oxygen-glucose deprivation (OGD) for 3 hours (hypoxia chamber with 95% N2 and 5% CO2, glucose-free serum-free medium) followed by reoxygenation for 12 hours (normoxia, complete medium) to simulate spinal cord ischemia-reperfusion injury. Pretreatment group received prostaglandin E1 for 2 hours after OGD and before reoxygenation. Inhibitor group received ML385 (Nrf2 inhibitor) for 2 hours after OGD, then prostaglandin E1 for 2 hours, followed by reoxygenation for 12 hours. After treatment, intracellular malondialdehyde, glutathione, and Fe2+ levels were measured; cell viability was assessed by CCK-8; immunofluorescence staining and western blot were used to detect ACSL4 and GPX4 expression; flow cytometry measured reactive oxygen species; western blot detected Nrf2 and HO-1 protein expression. (2) Animal experiment: 45 rats were randomly divided into three groups: sham group (n=15) underwent laparotomy without aortic occlusion; model group (n=15) underwent occlusion of abdominal aorta for 30 minutes followed by tail vein injection of saline, then reperfusion; pretreatment group (n=15) underwent occlusion for 30 minutes followed by tail vein injection of prostaglandin E1, then reperfusion. At 24 hours after reperfusion, motor function and neuronal injury were assessed by BBB score, inclined plane test, and Nissl staining; blood-spinal cord barrier integrity and microvascular density were evaluated by spinal cord water content, immunofluorescence staining of ZO-1, and CD34 immunohistochemistry; ferroptosis in spinal cord tissue was assessed by immunofluorescence, Prussian blue staining, western blot, and biochemical assays. RESULTS AND CONCLUSION: (1) Cell experiment: OGD/reoxygenation reduced cell viability, induced ferroptosis, and downregulated Nrf2 and HO-1 protein expression in rat spinal cord microvascular endothelial cells. Prostaglandin E1 pretreatment inhibited these effects; ML385 partially reversed the protective effect of prostaglandin E1. (2) Animal experiment: Prostaglandin E1 pretreatment alleviated motor dysfunction, neuronal injury, and blood-spinal cord barrier damage, improved microvascular density, and inhibited ferroptosis in spinal cord tissue after spinal cord ischemia-reperfusion injury. (3) These results indicate that prostaglandin E1 pretreatment protects against spinal cord ischemia-reperfusion injury by activating the Nrf2/HO-1 signaling pathway to inhibit ferroptosis in endothelial cells.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21299
BACKGROUND: Neurogenic heterotopic ossification frequently occurs within 1 to 3 months following spinal cord injury or traumatic brain injury, characterized by abnormal bone formation in periarticular soft tissues. The precise pathogenesis remains unclear, underscoring the urgent need for systematic research to inform clinical management. OBJECTIVE: To summarize recent advances in animal models of neurogenic heterotopic ossification and elucidate its underlying mechanisms, with a particular focus on the pathological differentiation of osteogenic precursor cells, remodeling of the local tissue microenvironment, and the interplay between neural regulation and neurogenic heterotopic ossification formation. METHODS: PubMed, CNKI, and SinoMed were searched from inception to January 2025. Chinese search terms included 'neurogenic heterotopic ossification, spinal cord injury, traumatic brain injury, heterotopic ossification'; English search terms included 'Neurogenic Heterotopic Ossification, spinal cord injury, Traumatic brain injury, ossification, heterotopic, Central nervous system'. Literature related to animal models and mechanisms of neurogenic heterotopic ossification was included to summarize key pathogenic processes. RESULTS AND CONCLUSION: The recruitment and aberrant osteogenic differentiation of osteogenic precursor cells (mainly fibro-adipogenic progenitors) are regulated by local microenvironmental factors such as hypoxia, inflammation, and angiogenesis. Neurotrophic factors, calcitonin gene-related peptide, and substance P promote aberrant ossification through neuro-immune interactions. Future research should construct a systematic molecular map, explore core signaling pathways, and develop novel targeted interventions to achieve early identification and individualized treatment of neurogenic heterotopic ossification, thereby improving patient outcomes.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21408
BACKGROUND: Interlocking intramedullary nail fixation is the "gold standard" for the treatment of femoral shaft fractures, and the difficulty of distal locking nail implantation has always been a difficult problem to solve. OBJECTIVE: By comparing the clinical effects of correction leverage technique and free-hand locking nail technique, it is further explained whether the correction leverage technique can be fast and accurate. The distal locking screw of femoral intramedullary nail was placed without direct X-ray radiation exposure, thereby solving the problem of difficult distal locking screw placement. METHODS: A total of 52 patients with femoral shaft fractures who had difficulty in distal locking screw placement during interlocking intramedullary nail fixation were enrolled from the Department of Orthopedics and Traumatology, Affiliated Traditional Chinese Medicine Hospital of Southwest Medical University from July 2022 to September 2024. They were divided into two groups according to the placement protocol: correction leverage group (26 cases) used the correction leverage technique for distal locking screw placement, and free-hand group (26 cases) used the traditional free-hand technique. The distal locking screw placement time, number of X-ray exposures, and first-attempt accuracy were compared between the two groups. RESULTS AND CONCLUSION: (1) The distal locking screw placement time in the correction leverage group was significantly shorter than that in the free-hand group (t=-4.136, P < 0.001). (2) The number of X-ray exposures in the correction leverage group was less than that in the free-hand group (t=-19.696, P < 0.001). (3) The first-attempt accuracy in the correction leverage group (100%) was higher than that in the free-hand group (71%), with a significant difference (χ2=5.253, P < 0.05). (4) These results indicate that compared with simple free-hand screw placement, the correction leverage technique has the advantages of faster locking, higher accuracy, and lower X-ray radiation. This technique does not require auxiliary equipment and has strong operability, and is worthy of further clinical validation and promotion for interlocking intramedullary nail fixation of femoral shaft fractures.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21391
BACKGROUND: In patients with Pauwels type III femoral neck fracture who are unable to undergo closed reduction, the traditional cannulated compression screw internal fixation method cannot effectively counteract the large shear force. To solve this problem, this study personalized an internally supported plate and used it in conjunction with the cannulated compression screw internal fixation protocol to improve treatment outcomes. OBJECTIVE: To compare the biomechanical performance of two internal fixation methods for Pauwels type III femoral neck fractures under gait loading: an "inverted triangle" arrangement of three screws and an "inverted triangle" arrangement of three screws combined with a medial support plate by finite element calculation. METHODS: Based on the CT scan data, inverse modeling was first performed using Mimics software to generate a point cloud model of the femur. Subsequently, the model was refined using Geomagic software to optimize its geometry and ensure the accuracy of the model. Finally, the processed model was imported into NX software to establish a femoral neck fracture model with a Pauwels angle of 70°. The mechanical and fatigue life results of the 3-nail [fully threaded (model 1), unthreaded (model 2)] model, 3-nail + personalized internal support plate [fully threaded (model 3), unthreaded (model 4)] model were computed based on the Ansys software for the gait loading. RESULTS AND CONCLUSION: (1) Under gait loading, the introduction of an internal support plate reduced the mean femoral stress compared with the 3-nail fixation approach, including a decrease in fracture and stump stresses of 6.6 MPa and 11.0 MPa, respectively; a decrease in displacement of 0.24 mm and 0.12 mm, respectively, and a reduction in relative displacement of the fracture surface. (2) The internal fixation method reduced the fatigue life of the bone system, and the addition of the internal support plate further reduced fatigue life. (3) The finite element analysis was sensitive to thread parameters, so the thread type and characteristics of screws should be considered in model construction. Compared with the 3-screw fixation alone, the addition of an internal support plate reduced stress and deformation levels, providing a more stable mechanical environment for bone healing. From the perspective of fatigue life, internal fixation reduced the life of the femoral system, and the more implants, the lower the life; the fixation scheme with the internal support plate had the lowest fatigue life. (4) This indicates that in clinical design of fixation schemes, the impact of implants on long-term healing outcomes should be fully considered, and the number of implants and fixation method should be comprehensively evaluated.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21381
BACKGROUND: Currently, the biomechanical differences between cervical rotation manipulation and cervical rotation-traction manipulation for the treatment of cervical radiculopathy have not been systematically elucidated. OBJECTIVE: To compare the biomechanical differences between cervical rotation manipulation and cervical rotation-traction manipulation in the treatment of cervical spondylotic radiculopathy caused by cervical disc herniation, and to provide a basis for the rational selection of manipulation in clinical practice. METHODS: A 27-year-old Asian male patient with cervical spondylotic radiculopathy caused by left posterior cervical disc herniation compressing the nerve root was recruited. The CT scan data of the skull and cervical spine were extracted to construct a finite element model of the head and full cervical spine. After model validation, the key parameters of cervical rotation manipulation and rotation-traction manipulation were loaded into the model, and the effects of the two manipulations on the stress of intervertebral disc, facet joints, spinal cord and nerve roots, disc displacement, and intervertebral foramen volume were compared. RESULTS AND CONCLUSION: (1) In terms of Von-Mise stress, the maximum stresses of cervical rotation manipulation on the annulus fibrosus, nucleus pulposus, and facet joints were 0.903, 0.139, and 2.186 MPa, respectively, which were significantly increased by 18%, 13%, and 30% compared with rotation-traction manipulation (0.765, 0.123, 1.682 MPa); while the maximum stress on the spinal cord and nerve roots was 2.547 MPa, which was 7% lower than that of rotation-traction manipulation (2.738 MPa). (2) In terms of displacement, the maximum forward displacement of the herniated side of the intervertebral disc by cervical rotation manipulation was 1.067 mm, which was 11.1% more than that of rotation-traction manipulation (0.960 mm). (3) In terms of intervertebral foramen volume changes, both manipulations increased the volume after implementation compared with before, with rotation manipulation increasing by 15.5% and rotation-traction manipulation increasing by 19.8%, the latter being more effective in expanding the intervertebral foramen volume. (4) It is suggested that cervical rotation manipulation has advantages in promoting the forward displacement of the herniated disc, but it produces higher stress on the intervertebral disc and facet joints, which may easily cause disc damage; rotation-traction manipulation will cause slightly higher stress on the spinal cord and nerve roots, but it can more effectively expand the intervertebral foramen volume and reduce the risk of disc structural damage. In clinical treatment, the advantages and disadvantages of the two manipulations should be carefully weighed and selected based on the patient's specific condition.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21316
BACKGROUND: The therapeutic potential of Toddalia asiatica in rheumatoid arthritis has garnered increasing attention, yet its mechanisms remain incompletely elucidated. OBJECTIVE: To investigate the underlying mechanisms of Toddalia asiatica in treating rheumatoid arthritis using bioinformatics combined with molecular dynamics simulation. METHODS: Active ingredients of Toddalia asiatica and their targets were retrieved. Drug targets were intersected with rheumatoid arthritis-related targets, followed by enrichment analysis of the overlapping genes. Molecular docking and molecular dynamics simulation were performed to validate the binding mechanisms of core active ingredients with key targets. RESULTS AND CONCLUSION: Through literature retrieval, 22 core active ingredients of Toddalia asiatica and their key targets against rheumatoid arthritis were identified. Enrichment analysis indicated that Toddalia asiatica may exert therapeutic effects by modulating disease-related signaling pathways (including cancer, infectious diseases, metabolic diseases, and cardiovascular diseases) as well as biological pathways related to metabolism, immunity, and inflammation. Meanwhile, the main components Dihydrochelerythrine and 8-Methoxychelerythrine specifically target phospholipase C gamma 2 (PLCG2) and mitogen-activated protein kinase 8 (MAPK8), respectively, suggesting that Toddalia asiatica may exert anti-rheumatoid arthritis effects through synergistic multi-pathway regulation.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21348
BACKGROUND: Platelets are important blood resources, yet in routine blood bank processes they are often filtered out along with white blood cells as medical waste. Optimizing whole blood separation processes to prepare platelet lysate products and exploring their applications in tissue engineering and regenerative medicine is of great value. OBJECTIVE: To optimize whole blood separation to prepare therapeutic-grade platelet lysate and to investigate the protective effect of platelet lysate on hypoxic injury of cardiomyocytes. METHODS: Platelets were isolated from 21 qualified whole blood units under closed blood bag and tubing conditions, and 21 platelet lysates were prepared by freeze-thawing. The mass concentration ranges of platelet-derived growth factor AA, platelet-derived growth factor BB, platelet-derived growth factor AB, vascular endothelial growth factor, epidermal growth factor, insulin-like growth factor 1, fibroblast growth factor, and transforming growth factor beta 1 in platelet lysates were measured using enzyme-linked immunosorbent assay kits. Bacterial contamination was assessed by colony culture method and mycoplasma contamination by PCR detection kit. A cardiomyocyte hypoxia model was established to evaluate the protective effect of platelet lysate on hypoxic injury. RESULTS AND CONCLUSION: (1) The mass concentration ranges of major growth factors and cytokines in platelet lysates were: platelet-derived growth factor AA 12.86-24.17 μg/L, platelet-derived growth factor BB 0.25-0.32 μg/L, platelet-derived growth factor AB 85.09-114.91 μg/L, vascular endothelial growth factor 10.57-58.37 μg/L, epidermal growth factor 0.43-0.69 μg/L, insulin-like growth factor 1 106-204.9 μg/L, fibroblast growth factor 0.03-0.06 μg/L, and transforming growth factor beta 1 124.17-192.38 μg/L. (2) Colony culture and mycoplasma detection results were negative. (3) Low volume fraction (1%) platelet lysate yielded the highest proliferation efficiency of cardiomyocytes; low volume fraction (1%) platelet lysate stimulated cardiomyocytes to produce high levels of superoxide dismutase and glutathione peroxidase to protect cardiomyocytes. This study established a method for preparing therapeutic-grade platelet lysate by optimizing the whole blood separation process, which can improve the utilization rate of blood resources. Platelet lysate has high levels of major growth factors and can significantly promote the repair of hypoxic injured cardiomyocytes.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21337
BACKGROUND: Myelination deficits are a core feature of white matter injury in preterm infants. In recent years, human umbilical cord mesenchymal stem cells have been applied in various animal models of brain injury, demonstrating the capacity to promote myelin repair. Elucidating the regulatory mechanisms by which human umbilical cord mesenchymal stem cells enhance neural myelination will contribute to optimizing therapeutic strategies and facilitating clinical translation. OBJECTIVE: To clarify the reparative effect of human umbilical cord mesenchymal stem cells on myelination disorders caused by maturation arrest of the oligodendrocyte lineage in neonatal rats with white matter injury. METHODS: Seventy-two 2-day-old Sprague-Dawley rats were randomly divided into sham operation group, white matter injury group, and human umbilical cord mesenchymal stem cell transplantation group (n=24 per group). A neonatal rat model of white matter injury was established by combining low-dose lipopolysaccharide with hypoxia-ischemia. On day 14 after modeling, pathological changes in white matter were observed by hematoxylin-eosin staining; the positive expression, protein and mRNA levels of oligodendrocyte lineage transcription factor 2, neural/glial antigen 2, and myelin basic protein were detected by immunohistochemistry, western blot, and real-time quantitative PCR. On day 28 after modeling, myelin formation was observed by Luxol fast blue staining, and spatial learning and memory ability were tested by Morris water maze. RESULTS AND CONCLUSION: On day 14 after modeling, hematoxylin-eosin staining showed that in the white matter injury group, a large number of cells degenerated and necrosed, and nerve fibers were arranged disorderly; in the human umbilical cord mesenchymal stem cell transplantation group, cell morphology was close to normal and nerve fibers were arranged relatively neatly. On day 14 after modeling, there was no statistically significant difference in the positive expression, protein and mRNA levels of oligodendrocyte lineage transcription factor 2 among groups (P > 0.05). Compared with the sham operation group, the positive expression, protein and mRNA levels of neural/glial antigen 2 were upregulated (P < 0.05), while those of myelin basic protein were downregulated (P < 0.05) in the white matter injury group. Compared with the white matter injury group, the positive expression, protein and mRNA levels of neural/glial antigen 2 were downregulated (P < 0.05), while those of myelin basic protein were upregulated (P < 0.05) in the human umbilical cord mesenchymal stem cell transplantation group. On day 28 after modeling, Luxol fast blue staining showed that compared with the sham operation group, myelin expression was decreased in the white matter injury group (P < 0.05); compared with the white matter injury group, myelin expression was increased in the human umbilical cord mesenchymal stem cell transplantation group (P < 0.05). On day 28 after modeling, Morris water maze results showed that compared with the sham operation group, the escape latency was prolonged and the number of platform crossings was decreased in the white matter injury group (P < 0.05); compared with the white matter injury group, the escape latency was shortened and the number of platform crossings was increased in the human umbilical cord mesenchymal stem cell transplantation group (P < 0.05); there was no statistically significant difference in average swimming distance among groups (P > 0.05). These findings indicate that human umbilical cord mesenchymal stem cells can promote the maturation of the oligodendrocyte lineage, repair myelination disorders, and improve cognitive function in neonatal rats with white matter injury.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21330
BACKGROUND: Studies showed that multiple myeloma microenvironment has the function of inducing mesenchymal stem cells to become senescent phenotype, while the effect of senescent bone marrow mesenchymal stem cells on multiple myeloma cells is rarely reported. OBJECTIVE: To investigate the effect of senescent bone marrow mesenchymal stem cells on the proliferation of multiple myeloma cells through paracrine galectin-3. METHODS: Bone marrow blood was collected from healthy donors, and bone marrow mesenchymal stem cells were extracted by Ficoll density gradient centrifugation and adherent purification. The third-generation bone marrow mesenchymal stem cells were taken and induced with 200 µmol/L hydrogen peroxide solution for 2 h, then cultured with L-DMEM complete medium for 24 h to construct a senescent bone marrow mesenchymal stem cell model. The model was identified by β-galactosidase staining and senescence gene P21 expression. RT-qPCR was used to detect the expression of galectin-3 in senescent bone marrow mesenchymal stem cells. The supernatant of senescent bone marrow mesenchymal stem cells was collected and concentrated by centrifugation to prepare conditioned medium, which was used to culture multiple myeloma cell line U266 for 24 h. CCK-8 was used to detect U266 cell proliferation, flow cytometry was used to detect U266 cell apoptosis, and RT-qPCR and western blot were used to detect BCL-2 protein and mRNA expression in U266 cells. Bone marrow from multiple myeloma patients and healthy individuals was collected, and galectin-3 levels were detected by ELISA. RESULTS AND CONCLUSION: After hydrogen peroxide induction, the number of β-galactosidase positive cells significantly increased, and the mRNA expression of P21 and galectin-3 was upregulated (P < 0.01). Compared with the control group, after culturing U266 cells with senescent bone marrow mesenchymal stem cell conditioned medium for 24 h, cell proliferation increased (P < 0.05), apoptosis rate decreased (P < 0.05), and BCL-2 protein and mRNA expression levels increased (P < 0.05). The level of galectin-3 in bone marrow of multiple myeloma patients was significantly higher than that of healthy individuals (P < 0.05). The results indicate that senescent bone marrow mesenchymal stem cells may promote the proliferation of multiple myeloma cells by upregulating BCL-2 expression through paracrine galectin-3.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21394
BACKGROUND: Systemic fatigue increases injury risk in individuals with functional ankle instability, while stop-jump cutting is a high risk for ankle injuries. The biomechanical mechanisms underlying non-anticipated stop-jump cutting during systemic exercise fatigue in this population remain unclear. OBJECTIVE: To quantify differences in kinematic and kinetic characteristics during non-anticipated stop-jump cutting before and after exercise fatigue between individuals with functional ankle instability and healthy controls, revealing the impact of exercise fatigue on stop-jump cutting in individuals with functional ankle instability. METHODS: Fifteen male participants with unilateral functional ankle instability and 15 healthy male controls were recruited. Kinematic (peak angles of ankle dorsiflexion, plantarflexion, inversion, knee flexion, knee varus, knee valgus, hip flexion, and hip abduction) and kinetic (joint stiffness of hip, knee, and ankle) parameters were collected during non-anticipated stop-jump cutting before and after exercise fatigue. Two-way repeated measures ANOVA was used to analyze peak joint angles and joint stiffness. Statistical parametric mapping (SPM) was further used to analyze the effects of fatigue on time-series data of ankle angle and ground reaction forces. RESULTS AND CONCLUSION: Kinematic results: Two-way ANOVA revealed significant main effects of fatigue and group-by-fatigue interactions for peak ankle inversion, knee flexion, knee valgus, and hip flexion angles (P < 0.05). Compared with pre-fatigue, peak ankle inversion increased in the functional ankle instability group after fatigue (P < 0.05), peak knee flexion increased in both groups (P < 0.05), and peak hip flexion increased in the healthy control group (P < 0.05). After fatigue, the functional ankle instability group showed smaller peak ankle inversion and hip flexion angles but larger peak knee valgus and knee flexion angles than the healthy control group (P < 0.05). SPM analysis revealed that ankle inversion/eversion angle was greater during 4%-18% of the cutting movement after fatigue in the functional ankle instability group (P < 0.05). Kinetic results: Two-way ANOVA revealed significant main effects of fatigue and group-by-fatigue interactions for hip, knee, and ankle joint stiffness (P < 0.05). Compared with pre-fatigue, hip and ankle stiffness decreased in the healthy control group (P < 0.05), while knee and ankle stiffness decreased in the functional ankle instability group (P < 0.05). SPM analysis showed that vertical ground reaction force was greater during 5%-16% of the cutting movement, and mediolateral ground reaction force was greater during 35%-49% of the movement after fatigue in the functional ankle instability group (P < 0.05). CONCLUSION: Exercise fatigue alters kinematic and kinetic characteristics during non-anticipated stop-jump cutting in individuals with functional ankle instability, particularly affecting knee and ankle stability and shock absorption. Fatigue reduces joint stiffness and control, increasing injury risk, especially during the initial and transitional phases of the cutting movement.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21449
BACKGROUND: Percutaneous coronary intervention stent implantation is primarily used to treat coronary artery stenosis. However, current multi-objective stent optimization methods are limited by sample size constraints, resulting in insufficient prediction accuracy when balancing key performance indicators such as support and compliance, hindering the effectiveness of stent optimization design. OBJECTIVE: To establish an innovative optimization framework for coronary stents based on a ensemble surrogate model. METHODS: A three-dimensional parametric model of the vascular stent was constructed, and a mechanical response database was established through finite element simulation. A dynamic weight fusion strategy was adopted to integrate the global optimization characteristics of the Kriging model and the local nonlinear representation advantages of the radial basis function model. A ensemble surrogate model was constructed based on 20 groups of initial samples, and the non-dominated sorting genetic algorithm-II was used to optimize the parameter space. RESULTS AND CONCLUSION: Experimental results demonstrated that the ensemble surrogate model exhibited significant advantages in the finite sample setting. The coefficient of determination for the inverse prediction of the radial stiffness of the stent reached 0.974 2, a 4.4% improvement compared to the single model, validating the efficient modeling capability of the ensemble surrogate model in the finite sample setting. The prediction accuracy of the stent's bending stiffness also improved by 4.4% compared to the single radial basis function surrogate model. After optimization, the stent performance achieved dual-objective synergistic optimization. The inverse radial stiffness of the stent in the ensemble surrogate model group was reduced by 13.92% and 9.57% compared to the Kriging model group and the single radial basis function surrogate model group, respectively. The bending stiffness of the stent was optimized by 0.38% and 2.56% compared to the Kriging model group and the single radial basis function surrogate model group, respectively. The proposed ensemble surrogate model breaks through the performance limitations of traditional single models, providing a low-cost, high-precision solution for the 'rigid-flexible' synergistic optimization of coronary stents.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21505
BACKGROUND: The Affiliated Hospital of Jiangxi University of Chinese Medicine has used traditional Chinese medicine (TCM) to treat children with cerebral palsy (CP) for over 20 years, but no analysis of medication patterns has been conducted. OBJECTIVE: To analyze TCM syndrome types and explore medication patterns for CP in children based on medical records and literature. METHODS: An evidence-based retrieval strategy was used to search and manage literature and medical records on TCM treatment for CP in children. Bibliometric methods were applied to mine and analyze data characteristics. VOSviewer software was used to create visual knowledge maps. IBM SPSS Modeler software was used for association rule analysis of TCM drugs. Radar chart method was used to analyze the four natures and five flavors of drugs. RESULTS AND CONCLUSION: A total of 503 medical records and 90 articles were included. Syndrome analysis showed that the main TCM syndrome type of CP in children was liver-kidney deficiency. Intervention analysis showed that external therapy was most frequently used, and among oral medications, drugs for nourishing liver and kidney were most common. Medication pattern analysis showed that among the top 20 drugs by frequency in both medical records and literature, 12 (60%) were the same. The drug pairs Shanyao (Rhizoma Dioscoreae) and Fuling (Poria), and Shudi (Radix Rehmanniae Preparata) and Fuling (Poria) had high support and confidence above 82.50%, indicating significant association. Among all TCM drugs for CP in children, the nature was mainly warm, followed by neutral and cold; the flavor was mainly sweet, followed by bitter and pungent; the meridian tropism was mainly liver and kidney meridians, followed by spleen, heart, and lung meridians. Most drugs in these formulas were non-toxic. The results indicate that CP in children is mainly characterized by liver-kidney deficiency, and the formulas used in clinical practice and related clinical research are mostly for nourishing liver and kidney, among which Liuwei Dihuang Pill and its derivative formulas appear most frequently. The drug pairs with the highest frequency and reliability are Shudi and Fuling, and Shanyao and Fuling. The nature and flavor of drugs are mainly warm, sweet, and attributed to liver and kidney meridians.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21492
BACKGROUND: Previous studies from our group have shown that triptolide exerts protective effects on nerve cells and alleviates symptoms of neurodegenerative diseases. However, whether it acts by improving mitochondrial dynamic abnormalities requires further investigation. OBJECTIVE: To explore the effect and mechanism of triptolide in regulating the mitochondrial fusion-fission balance to mitigate hydrogen peroxide (H₂O₂)-induced apoptosis in SH-SY5Y cells. METHODS: Human neuroblastoma SH-SY5Y cells were cultured and divided into three groups: control group, model group (200 μmol/L H₂O₂), and triptolide group (2.5 nmol/L triptolide + 200 μmol/L H₂O₂). After 24 hours of intervention, oxidative stress markers (superoxide dismutase activity and malondialdehyde levels), mitochondrial membrane potential, and apoptosis levels were measured. Western blot was used to detect the expression of apoptosis-related proteins, mitochondrial dynamics-related proteins, and respiratory chain-related proteins. Immunofluorescence staining was used to detect the expression of phosphorylated dynamin-related protein 1, optic atrophy protein 1, cytochrome C oxidase 1, and ATP synthase F1 subunit alpha. RESULTS AND CONCLUSION: Compared with the control group, the model group showed significantly decreased superoxide dismutase activity, mitochondrial membrane potential, anti-apoptotic protein Bcl-2, mitochondrial fusion proteins 1 and 2, optic atrophy protein 1, and oxidative phosphorylation complex proteins (NADH dehydrogenase [ubiquinone] iron-sulfur protein 8, ubiquinol-cytochrome c reductase core protein 2, cytochrome c oxidase 1, succinate dehydrogenase B, ATP synthase F1 subunit alpha) (P < 0.05). Meanwhile, malondialdehyde levels, pro-apoptotic proteins Bax and Caspase-3, mitochondrial fission protein 1, phosphorylated dynamin-related protein 1 expression, and apoptosis rate were significantly increased (P < 0.05). Compared with the model group, triptolide intervention reduced malondialdehyde levels, increased superoxide dismutase activity and mitochondrial membrane potential, promoted fusion protein expression, inhibited fission protein expression, increased oxidative phosphorylation complex protein levels, and decreased apoptosis rate (P < 0.05). These results confirm that triptolide can regulate mitochondrial dynamic imbalance to alleviate H₂O₂-induced apoptosis in SH-SY5Y cells.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21488
BACKGROUND: Mangiferin, a major bioactive compound derived from mango trees, is widely present in various traditional Chinese medicinal herbs and exhibits multiple biological functions including antibacterial, cholesterol-lowering, and anti-allergic effects. Existing studies have suggested that mangiferin may prevent and treat osteoarthritis pain. However, its specific mechanism of action remains unclear to date. OBJECTIVE: To systematically investigate the key targets and potential mechanisms of mangiferin in the treatment of osteoarthritis by integrating gene expression omnibus (GEO) microarray data analysis, network pharmacology, and molecular docking techniques, and to validate the findings in a rat model. METHODS: First, GEO microarray data were mined to identify potential therapeutic targets for osteoarthritis. Next, professional databases were integrated to predict the targets of mangiferin, and target information related to osteoarthritis was collected. A Venn diagram was generated using the Weishengxin platform, a protein-protein interaction network was constructed based on the STRING database, and Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses were performed. Cytoscape 3.8.0 software was used to construct a drug-target-pathway-disease network, and molecular docking analysis and visualization were performed using the CBDOCK2 online docking platform. A rat model of osteoarthritis was established by anterior cruciate ligament transection of the left knee joint, and different concentrations of mangiferin were administered to observe and record the therapeutic effects. RESULTS AND CONCLUSION: A total of 144 potential targets of mangiferin were identified from multiple databases. Protein-protein interaction network analysis revealed important targets including interleukin-6, tumor necrosis factor, and nuclear factor kappa B1. Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis showed that 235 signaling pathways might be involved, including lipid and atherosclerosis-related pathways, advanced glycation end products-receptor, hypoxia-inducible factor 1, and estrogen, which are closely related to inflammation. In animal experiments, after 4 weeks of intervention with 40 μmol/L mangiferin, there was no significant difference in hindlimb weight-bearing compared with the sham-operated group. These findings suggest that mangiferin may exert therapeutic effects on osteoarthritis through a multi-target, multi-pathway mode of action, providing a new strategy and theoretical support for the treatment of osteoarthritis.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21483
BACKGROUND: Oxidative stress is one of the potential factors contributing to muscle atrophy following anterior cruciate ligament injury. Alleviating skeletal muscle fatigue facilitates proprioceptive recovery, thereby accelerating rehabilitation after anterior cruciate ligament injury. Improving mitochondrial function helps mitigate skeletal muscle fatigue-related damage. OBJECTIVE: To verify that electroacupuncture at different frequencies alleviates skeletal muscle oxidative stress damage and improves mitochondrial function in rabbits, thereby reducing skeletal muscle fatigue, restoring proprioceptive function, and accelerating rehabilitation following anterior cruciate ligament injury. METHODS: Twenty-four healthy New Zealand rabbits were randomly divided into blank group, model group, low-frequency electroacupuncture group, and high-frequency electroacupuncture group, with 6 rabbits in each group. The model group, low-frequency electroacupuncture group, and high-frequency electroacupuncture group were used to construct a knee anterior cruciate ligament injury model. In the low-frequency and high-frequency electroacupuncture groups, electroacupuncture was applied to the acupoints Xuehai and Liangqiu on the affected knee joint 7 days after modeling. The blank and model groups were only grasped and fixed without electroacupuncture intervention, once daily for 21 consecutive days. After intervention, ELISA was used to detect the levels of superoxide dismutase, succinate dehydrogenase, and malondialdehyde in the quadriceps femoris; western blot was used to detect the protein expression levels of silent information regulator 2-related enzyme 1, peroxisome proliferator-activated receptor gamma coactivator 1 alpha, and mitochondrial transcription factor A in skeletal muscle tissue, as well as the mRNA expression of silent information regulator 2-related enzyme 1, peroxisome proliferator-activated receptor gamma coactivator 1 alpha, nuclear respiratory factor 1, and mitochondrial transcription factor A. RESULTS AND CONCLUSION: After anterior cruciate ligament injury, the level of superoxide dismutase in the quadriceps femoris of rabbits increased, the level of succinate dehydrogenase decreased, and the concentration of malondialdehyde increased; the protein expressions of silent information regulator 2-related enzyme 1, peroxisome proliferator-activated receptor gamma coactivator 1 alpha, and mitochondrial transcription factor A decreased; the mRNA expressions of silent information regulator 2-related enzyme 1, peroxisome proliferator-activated receptor gamma coactivator 1 alpha, nuclear respiratory factor 1, and mitochondrial transcription factor A were significantly downregulated. After electroacupuncture intervention, the concentration of malondialdehyde in the affected quadriceps femoris decreased, the activities of superoxide dismutase and succinate dehydrogenase increased, and the protein expressions of silent information regulator 2-related enzyme 1, peroxisome proliferator-activated receptor gamma coactivator 1 alpha, and mitochondrial transcription factor A increased; the mRNA expressions of mitochondrial biogenesis-related genes such as silent information regulator 2-related enzyme 1, peroxisome proliferator-activated receptor gamma coactivator 1 alpha, nuclear respiratory factor 1, and mitochondrial transcription factor A increased, and the low-frequency electroacupuncture group was superior to the high-frequency electroacupuncture group. These results indicate that electroacupuncture can reduce oxidative stress damage in skeletal muscle after anterior cruciate ligament injury by increasing the contents of superoxide dismutase and succinate dehydrogenase and decreasing the content of malondialdehyde; and improve mitochondrial function by regulating the expression of proteins related to the silent information regulator 2-related enzyme 1/peroxisome proliferator-activated receptor gamma coactivator 1 alpha signaling pathway and mitochondrial biogenesis-related genes, thereby accelerating rehabilitation after anterior cruciate ligament injury.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21554
BACKGROUND: Studies have shown that circulating inflammatory proteins and immune cells are associated with rheumatoid arthritis, but the causal relationship is unclear. OBJECTIVE: To explore the causal relationships between circulating inflammatory proteins and rheumatoid arthritis mediated by immune cells. METHODS: We downloaded data on circulating inflammatory proteins and immune cell phenotypes from the GWAS Catalog database (a publicly accessible database jointly established and maintained by the National Human Genome Research Institute and the European Bioinformatics Institute), and genome-wide association study data for rheumatoid arthritis from the FinnGen database (a genomics project resulting from collaboration between Finnish research institutions, biobanks, and international industry partners, also publicly accessible). Two-step Mendelian randomization analyses were performed: inverse variance weighting was used to assess the causal effects of 91 circulating inflammatory proteins and 731 immune cell phenotypes on rheumatoid arthritis risk, supplemented by MR-Egger, weighted median, weighted mode, simple mode, and sensitivity analyses. The mediating role of identified immune cells in the relationship between circulating inflammatory proteins and rheumatoid arthritis was evaluated. RESULTS AND CONCLUSION: Inverse variance weighting analysis showed that four circulating inflammatory proteins were significantly associated with rheumatoid arthritis risk, of which one was a risk factor and three were protective factors. Forty-six immune cell phenotypes were significantly associated with rheumatoid arthritis, of which 20 were risk factors and 26 were protective factors. Reverse Mendelian randomization analysis found no causal association between rheumatoid arthritis and the four identified circulating inflammatory proteins. Sensitivity analyses revealed no significant heterogeneity or horizontal pleiotropy. Further mediation analysis showed that CD19 on IgD- CD38br partially mediated the causal effect of interleukin-18 (β=0.064, OR=1.066, P=0.044) on rheumatoid arthritis, with a mediation effect of 0.004, a mediation proportion of 5.7%, and a direct effect of 0.060. The results reveal causal associations between circulating inflammatory proteins and immune cells with rheumatoid arthritis, and identify that CD19 on IgD- CD38br partially mediates the causal relationship between interleukin-18 and rheumatoid arthritis. For the Chinese biomedical research field, reference can be made to the integrated analysis framework of international multi-omics platforms and cross-ethnic cohort data to construct a combined database of epigenomics, proteomics, and metabolomics specific to the Chinese population, revealing the molecular regulatory networks underlying complex diseases, and facilitating disease subtyping and early diagnostic biomarker development. By learning from transnational collaborative research mechanisms, establish natural population cohorts of multiple ethnicities and regions in China, systematically analyze the impact of environmental exposure and gene interactions on health, and provide scientific evidence for formulating localized disease prevention strategies.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21529
BACKGROUND: Knee osteoarthritis is an age-related disease, and aging is closely related to its occurrence and development. Chondrocyte senescence plays a crucial role in the pathological progression of knee osteoarthritis. OBJECTIVE: To establish a stable induced knee osteoarthritis model in SD rats. METHODS: (1) Animal experiment: Forty Sprague-Dawley rats were randomly divided into four groups: blank control group (no modeling), D-galactose group (intra-articular injection of D-galactose solution once a week for 2 months), anterior cruciate ligament transection (ACLT) group (ACLT to establish knee osteoarthritis model), and D-galactose+ACLT group (ACLT followed by intra-articular injection of D-galactose solution once a week for 2 months). One week after modeling, all rats underwent running exercise for 30 min every other day. At 4 and 8 weeks after modeling, behavioral tests (Lequesne MG score) were performed, and then samples were collected for detection of inflammatory factors in synovial fluid, histopathological morphology of knee cartilage, transmission electron microscopy observation, and immunohistochemical staining of type II collagen and aggrecan. (2) Cell experiment: At 4 and 8 weeks after modeling, knee chondrocytes were isolated from each group for flow cytometry cell cycle analysis, β-galactosidase staining, and γ-H2AX immunofluorescence staining. RESULTS AND CONCLUSION: (1) Animal experiment: At 8 weeks after modeling, Lequesne MG scores in the three model groups were higher than those in the blank control group (P < 0.05), and the score in the D-galactose+ACLT group was higher than that in the D-galactose and ACLT groups (P < 0.05). At 4 and 8 weeks, levels of interleukin-1β, interleukin-6, and tumor necrosis factor-α in synovial fluid were higher in the three model groups than in the blank control group (P < 0.05), and these levels were higher in the D-galactose+ACLT group than in the D-galactose and ACLT groups (P < 0.05). Hematoxylin-eosin and safranin O/fast green staining and transmission electron microscopy at 4 and 8 weeks showed that cartilage damage and chondrocyte mitochondrial damage were more severe in the D-galactose+ACLT group than in the D-galactose and ACLT groups. Immunohistochemical staining showed that the expression of type II collagen and aggrecan was highest in the blank control group, and lowest in the D-galactose+ACLT group among the model groups. (2) Cell experiment: At 4 and 8 weeks, the proportion of chondrocytes in G0/G1 phase was higher, and the proportions in S and G2/M phases were lower in the D-galactose+ACLT group than in the other three groups (P < 0.05). The positive rate of β-galactosidase staining and γ-H2AX immunofluorescence intensity were higher in the D-galactose+ACLT group than in the other three groups (P < 0.05). (3) These results indicate that the D-galactose+ACLT method can establish an SD rat model of aging-related knee osteoarthritis, which can better simulate the pathological state of aging and degeneration in knee osteoarthritis.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21591
BACKGROUND: Pharmacodynamic characteristics and mechanisms of action of icariin in combating osteoporosis gradually gain recognition within the academic community. Related basic research and clinical translation efforts are increasingly becoming the focal point of research. OBJECTIVE: To summarize the research progress of icariin on anti-osteoporosis. METHODS: China National Knowledge Infrastructure (CNKI) and PubMed databases were searched for relevant literature. Chinese and English search terms included “icariin, osteoporosis, Chinese medicine compound, pathogenesis, signal path, BMSCs, osteoblast, osteoclast.” Based on inclusion criteria, 90 articles were ultimately included in the review. RESULTS AND CONCLUSION: Icariin treatment increased alkaline phosphatase activity and induced the expression of core binding factor α1, bone morphogenetic protein 2, and bone morphogenetic protein 4 in bone marrow mesenchymal stem cells in a dose-dependent manner. Icariin promoted fracture healing by increasing serum levels of osteocalcin, bone-specific alkaline phosphatase, N-terminal peptide of type I collagen, C-terminal peptide of type I collagen, and tartrate-resistant acid phosphatase 5b, thereby enhancing osteocalcin secretion at the fracture site. Icariin promoted proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells in ovariectomized osteoporotic rats by upregulating alkaline phosphatase and osteocalcin levels and inhibiting the expression of Notch-1, CBF1, and Jagged-1 proteins in the Notch pathway, thus achieving prevention and treatment of osteoporosis. Icariin regulates bone metabolism through multiple signaling axes including Wnt/β-catenin, mitogen-activated protein kinase, phosphatidylinositol 3-kinase/protein kinase B, osteoprotegerin/receptor activator of nuclear factor-κB ligand/receptor activator of nuclear factor-κB, and Notch. Among these, the Wnt/β-catenin pathway and the osteoprotegerin/receptor activator of nuclear factor-κB ligand/receptor activator of nuclear factor-κB axis constitute the core regulatory mechanism, modulating the osteoblast-osteoclast dynamic balance through synergistic interactions. Icariin can influence the biological behavior of osteoblasts, osteoclasts, and bone marrow mesenchymal stem cells through multidimensional interventions including regulation of mRNA expression modifications, inhibition of oxidative stress, and improvement of the inflammatory microenvironment.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21593
BACKGROUND: Intervertebral disc degeneration is a core pathological mechanism of discogenic diseases, characterized by an imbalance in extracellular matrix metabolism. Tissue inhibitors of metalloproteinases, as endogenous antagonists of matrix metalloproteinases, play a crucial role in regulating extracellular matrix homeostasis, but the specific functions of subtypes, signaling pathway interactions, and epigenetic regulatory mechanisms have not been systematically clarified. OBJECTIVE: To review the expression changes, functional heterogeneity, and regulatory networks of tissue inhibitors of metalloproteinases in intervertebral disc degeneration, focusing on the molecular mechanisms and signaling pathways in oxidative stress, mechanical load, and inflammatory microenvironment, and to evaluate the translational potential of gene therapy strategies based on tissue inhibitors of metalloproteinases. METHODS: The first author searched PubMed, Web of Science, Embase, CNKI, Wanfang and other databases from inception to March 2025. Chinese search terms included '椎间盘退变，椎间盘退行性变，椎间盘退化，金属蛋白酶类组织抑制剂，信号通路', and English search terms included 'Tissue Inhibitor of Metalloproteinases, Tissue Inhibitor of Metalloproteinase, TIMPs, Intervertebral disc degeneration, Disc degeneration, Degenerative Disc Disease, Degenerative Intervertebral Discs'. Finally, 76 eligible articles were included for review. RESULTS AND CONCLUSION: (1) Subtype functions of tissue inhibitors of metalloproteinases: TIMP1 exhibits dual regulation (early protection/late depletion); TIMP2 maintains extracellular matrix homeostasis by inhibiting matrix metalloproteinase activity, and its abnormal expression can activate pro-apoptotic signaling pathways (e.g., miR-185-5p/MMP2 axis and inflammatory factor-mediated MMP/TIMP imbalance); TIMP3 exerts multi-dimensional protective effects by inhibiting matrix metalloproteinase activity, tumor necrosis factor-alpha converting enzyme/tumor necrosis factor-alpha axis, and angiogenesis; TIMP4 is regulated by miR-155-5p/fibroblast growth factor 2 and participates in extracellular matrix homeostasis. (2) Epigenetic reprogramming mechanisms: Abnormal mechanical stress degrades TIMP3 mRNA through the WTAP/YTHDF2-m6A axis, while miR-222 targets TIMP3 to synergistically accelerate extracellular matrix degradation. (3) Multi-modal therapeutic strategies: Photobiomodulation (wavelength-specific regulation of TIMP/MMP), stem cell exosomes (miR-199a/GREM1 axis), and irisin intervention can remodel matrix metabolic balance. This review summarizes the theoretical framework of 'functional network imbalance of tissue inhibitors of metalloproteinases', revealing its multi-level regulatory characteristics as a core driver of intervertebral disc degeneration, and providing a theoretical basis for developing precise therapies targeting epigenetic modifications and mechano-biological coupling interventions.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2026045
Colorectal cancer (CRC) remains a leading cause of cancer-related mortality, necessitating the discovery of novel therapeutic agents. Here, we report a natural small molecule, 2-dihydroailanthone (2-DAIL), as a promising candidate for CRC treatment. First, our results demonstrate that 2-DAIL exhibits significant anti-CRC activity in vitro and in vivo. Then, we find that 2-DAIL directly binds to integrin alpha-3 (ITGA3) revealed by stable isotope labeling by amino acids in cell culture coupled with thermal proteome profiling (SILAC-TPP). Additionally, the RNA sequencing data obtained from CRC cells and tumor tissues suggest that 2-DAIL blocks the PI3K/AKT signaling pathway mediated by ITGA3 inhibition. Collectively, 2-DAIL exerts its anti-CRC effects, at least partially, by binding to and inhibiting the function of ITGA3, thereby blocking the activation of the PI3K/AKT signaling pathway, which leads to CRC cell growth inhibition. Our study provides a promising drug candidate for the treatment of CRC and suggests the potential of 2-DAIL in treating other diseases linked to ITGA3 dysfunction.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2026020
The insulin receptor (IR) is central to the regulation of glucose and lipid metabolism. Although insulin is its primary ligand, insulin-like growth factors I and II (IGF-I and IGF-II) also engage IR, albeit with reduced affinity. The structural basis of cooperative ligand binding, however, has remained poorly understood. Here, we report cryo-Electron Microscopy (cryo-EM) structures of IR in complex with insulin, IGF-I, and IGF-II, revealing that all three ligands engage the receptor at overlapping binding sites and can induce a conserved T-shaped quaternary assembly involving four ligand molecules at site 1/1′ and site 2/2′. Despite this shared overall architecture, distinct ligand-specific conformational changes are observed. Notably, IGF-I and IGF-II adopt different binding sequence at site 1 and site 2 compared to insulin, suggesting unique interaction dynamics. These structural insights highlight divergent mechanisms of ligand recognition and cooperative binding, providing a deeper understanding of hormone-induced conformational modulation of the IR.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2025047
Systemic lupus erythematosus (SLE) is a prototypic autoimmune disease with complex pathogenesis historically attributed to adaptive immunity. Emerging data implicate neutrophils in immune dysregulation and organ damage. This study investigates neutrophil senescence dynamics in SLE. We identified a significantly elevated proportion of CXCR4hiCD62Llo senescence-like neutrophils in peripheral blood of SLE patients versus healthy donors. Increased senescence-like neutrophil numbers positively correlated with SLE disease activity and autoantibody production. Functionally, senescence-like neutrophils from SLE patients exhibited impaired suppression of proinflammatory activity in natural killer (NK) cells and CD4+ T cells. Mechanistically, these cells may exert immunosuppressive effects via reactive oxygen species (ROS) production under physiological conditions. Our results position senescence-like neutrophils as candidate biomarkers for SLE disease activity. The compromised immunosuppressive function of these cells offers a new perspective on SLE pathophysiology and may inform development of novel therapies. Limitations include small sample size and heterogeneous treatment backgrounds, necessitating further validation. Future studies will address NET release and potential subset markers.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2025233
This corrigendum addresses inaccuracies in three figure panels from the original article (Acta Biochim Biophys Sin 54: 1587–1598, doi: 10.3724/abbs.2022150). The authors identified that Figure 2D was mislabeled during preparation, Figure 4A contained an incorrect image due to a processing error, and Figure 5A was mistakenly replaced during final compilation. Corrected versions of these panels are provided. The authors confirm that these errors are confined to figure presentation and do not affect the underlying data, statistical analyses, or the main conclusions of the study. The original research demonstrated that caveolin-1 (CAV-1)-deficient fibroblasts promote migration, invasion, and stemness in breast cancer cells (BCCs) via activation of the TGF-β/Smad signaling pathway. Key experimental methods included Western blot for CAV-1 expression, scratch wound healing assays for migration, Transwell assays for migration and invasion, immunofluorescence and ELISA for TGF-β1 detection, and Western blot for EMT/stemness markers. Statistical significance was set at P < 0.05. This corrigendum ensures the accuracy of the scientific record and maintains the integrity of the reported findings.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2025048
Cervical cancer remains a leading cause of cancer-related mortality among women, with high-risk HPV-positive cases constituting 99% of instances. Despite wild-type p53 expression, its tumor-suppressive function is crippled by E6AP-mediated ubiquitination and HDAC6-driven deacetylation, resulting in rapid degradation and low steady-state levels. This study evaluates a combinatorial strategy employing the natural product withaferin A (WA) and the HDAC6 inhibitor ricolinostat (RIC) to simultaneously target p53 ubiquitination and acetylation in HeLa, SiHa, and Caski cervical cancer cells. Dose-response CCK-8 assays established that both agents inhibit proliferation in a dose-dependent manner. Combination treatment significantly reduced cell viability compared to monotherapies, with CompuSyn analysis yielding a combination index (CI) below 1, confirming synergy. Colony formation assays further demonstrated a marked decrease in clonogenic survival. Mechanistically, WA disrupted the p53-E6AP interaction, reducing p53 ubiquitination, while RIC inhibited HDAC6-mediated deacetylation, increasing p53 acetylation. The dual treatment stabilized p53, as evidenced by extended half-life in cycloheximide chase assays. These findings suggest that concurrent modulation of p53 post-translational modifications via WA and RIC offers a potent therapeutic avenue for cervical cancer, meriting further preclinical development.
Chinese Journal of Tissue Engineering Research•2025•DOI: 10.12307/2025.20219
BACKGROUND: Adeno-associated virus (AAV) gene therapy has been proven to be reliable and safe for the treatment of osteoarthritis in recent years. However, given the complexity of osteoarthritis pathogenesis, single gene manipulation for the treatment of osteoarthritis may not produce satisfactory results. Previous studies have shown that nuclear factor κB could promote the inflammatory pathway in osteoarthritic chondrocytes, and bone morphogenetic protein 4 (BMP4) could promote cartilage regeneration. OBJECTIVE: To test whether combined application of AAV-p65shRNA and AAV-BMP4 will yield the synergistic effect on chondrocytes regeneration and osteoarthritis treatment. METHODS: Viral particles containing AAV-p65-shRNA and AAV-BMP4 were prepared. Their efficacy in inhibiting inflammation in chondrocytes and promoting chondrogenesis was assessed in vitro and in vivo by transfecting AAV-p65-shRNA or AAV-BMP4 into cells. The experiments were divided into five groups: PBS group; osteoarthritis group; AAV-BMP4 group; AAV-p65shRNA group; and BMP4-p65shRNA 1:1 group. Samples were collected at 4, 12, and 24 weeks postoperatively. Tissue staining, including safranin O and Alcian blue, was applied after collecting articular tissue. Then, the optimal ratio between the two types of transfected viral particles was further investigated to improve the chondrogenic potential of mixed cells in vivo. RESULTS AND CONCLUSION: The combined application of AAV-p65shRNA and AAV-BMP4 together showed a synergistic effect on cartilage regeneration and osteoarthritis treatment. Mixed cells transfected with AAV-p65shRNA and AAV-BMP4 at a 1:1 ratio produced the most extracellular matrix synthesis (P < 0.05). In vivo results also revealed that the combination of the two viruses had the highest regenerative potential for osteoarthritic cartilage (P < 0.05). In the present study, we also discovered that the combined therapy had the maximum effect when the two viruses were administered in equal proportions. Decreasing either p65shRNA or BMP4 transfected cells resulted in less collagen II synthesis. This implies that inhibiting inflammation by p65shRNA and promoting regeneration by BMP4 are equally important for osteoarthritis treatment. These findings provide a new strategy for the treatment of early osteoarthritis by simultaneously inhibiting cartilage inflammation and promoting cartilage repair.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2025143
Vascular calcification (VC) is an independent risk factor for cardiovascular morbidity and mortality, characterized by hydroxyapatite deposition in arterial walls, leading to increased stiffness, decreased compliance, and plaque rupture. No clinically acknowledged therapy reverses VC. Dihydrocapsaicin (DHC), the primary pungent capsaicinoid in chili peppers, exhibits analgesic, anticancer, anti-inflammatory, antioxidant, and anti-obesity properties. Using the Comparative Toxicogenomics Database, we identified 20 experimental target genes of DHC, including ATF4, CASP3, CASP4, CASP7, CAT, CDKN1A, CYP1A2, CYP2C19, CYP2C9, CYP2D6, CYP2E1, DDIT3, EIF2S1, ERN1, HSPA5, IGF1, MAP1LC3A, MAPK1, MAPK3, and TP53. Chemical-phenotype analysis revealed associations with apoptotic processes and autophagy. In a human vascular smooth muscle cell (hVSMC) calcification model induced by 1.2 μL of 100 mM CaCl2 in α-MEM basal medium, co-treatment with DHC (0.5, 2, or 8 μL of 4 mM solution) for 3–6 h significantly inhibited calcium deposition, as quantified by Alizarin Red staining and ImageJ analysis. These findings suggest that DHC modulates VC through mechanisms involving cell death, endoplasmic reticulum stress, and calcium signaling, highlighting its potential as a therapeutic agent for VC.