Stem Cell Research & Therapy•2026•DOI: 10.1186/s13287-026-04913-w
Background Extracorporeal cardiac shock wave (ECSW) therapy enhances the function of endothelial colony-forming cells (ECFCs), but whether it can serve as a preconditioning strategy to enhance myocardial infarction (MI) therapy remains unclear. This study investigated the efficacy and mechanism of intravenously delivered ECSW-preconditioned ECFCs (SW-ECFCs) in a rat MI model. Methods ECFCs were isolated from the bone marrow of ApoE-/- rats and fully characterized. RNA sequencing of control ECFCs versus SW-ECFCs revealed significant enrichment of the PI3K/AKT pathway. We therefore performed a series of in vitro functional assays on these cells, including Transwell migration, Matrigel tube formation, CCK-8 proliferation, flow cytometric apoptosis analysis, and VEGF-A ELISA. The role of the PI3K/AKT pathway was interrogated using the inhibitor LY294002. Subsequently, an acute MI model was established in ApoE-/- rats via left anterior descending coronary artery ligation. Rats were randomized into four groups: MI+PBS, MI+ECFCs, MI+SW-ECFCs, and MI+LY294002-pretreated SW-ECFCs (LY-SW-ECFCs), with sham-operated rats as controls. Comprehensive evaluations included echocardiography, serum injury biomarkers, TTC, and histopathological (H&E, Masson) staining, immunohistochemical detection of cardiomyocyte apoptosis and p-eNOS, immunofluorescence assessment of ECFC homing and vascular markers (CD31, α-SMA, VEGF-A), tissue/plasma nitric oxide measurement, and Western blot analysis of PI3K/AKT signaling proteins. Results Transcriptomic analysis revealed significant enrichment of the PI3K/AKT pathway in SW-ECFCs. Functionally, ECSW enhanced ECFCs migration, tube formation, proliferation, and VEGF-A secretion, while reducing apoptosis;
Stem Cell Research & Therapy•2026•DOI: 10.1186/s13287-026-05061-x
Background Osteoporosis (OP) is a systemic bone disease characterized by damage to bone strength, leading to increased bone fragility and fracture risk. Cadherin 19 (CDH19) is located on chromosomes 18q22-q23, and 18q deletion is associated with terminal deletion diseases, including foot/hand deformities. However, the role of CDH19 in bone remains undefined. Methods A conditional knockout mouse model of the CDH19 gene was constructed using the Cre-loxP system, and the bone mass and bone morphology in mice were investigated using microCT and histological staining. Osteoblasts were isolated and cultured from wild-type and CDH19 knockout mice. Cell proliferation and differentiation were explored through EdU labeling, qPCR, alkaline phosphatase (ALP)/ alizarin red S (ARS) staining, and Western blot assays. The expression of genes altered in CDH19 gene knockout osteoblast was checked by RNA sequencing (RNA-seq), and subsequently confirmed by immunofluorescence and Western blot. Results We found that CDH19 could maintain the normal proliferation and differentiation in osteoblasts. After knocking out the CDH19 gene, the abilities of proliferation and osteogenesis were significantly inhibited in osteoblasts. Moreover, the bone mass of CDH19 knockout mice was significantly reduced, characterized by decreases in bone density, trabecular number, and bone volume fraction. The RNAseq analysis and western blot showed the PI3K/AKT signaling pathway was significantly inhibited in osteoblasts with CDH19 deletion. Furthermore, we demonstrated that administration of PI3K/AKT signaling pathway agonist 740Y-P partially alleviated the inhibition of osteogenic differentiation caused by CDH19 deletion in vitro and in vivo. Conclusion This study demonstrated that CDH19 regulated osteogenic differentiation by modulating the PI3K/AKT signaling pathway in osteoblasts. CDH19 may become a novel target for the treatment of bone diseases.
Stem Cell Research & Therapy•2026•DOI: 10.1186/s13287-026-04902-z
Background Inherited cardiomyopathy (ICM) is a genetic disorder characterized by abnormal myocardial structure and function, often progressing to heart failure. FHOD3, a member of the Formin gene family, plays a crucial role in cardiomyocyte cytoskeletal organization. Mutations in FHOD3 have been associated with various cardiomyopathies, including hypertrophic cardiomyopathy (HCM), dilated cardiomyopathy (DCM) and left ventricular noncompaction (LVNC). However, the molecular mechanisms underlying FHOD3 deficiency-induced cardiomyopathy remain elusive. Methods A FHOD3 knockout (FHOD3-/-) human embryonic stem cell (hESC) line was generated using the CRISPR/Cas9 system and subsequently differentiated into cardiomyocytes (hESC-CMs). Sarcomere structure, calcium handling, mitochondrial function, and contractility were evaluated via immunofluorescence, electron microscopy, Seahorse metabolic analysis, and high-definition video analysis, respectively. Transcriptomic sequencing was performed to identify differentially expressed genes and enriched pathways. Results FHOD3-deficient hESC-CMs exhibited marked sarcomere disorganization and degradation, impaired calcium handling and compromised mitochondrial function, ultimately leading to reduced contractility. Transcriptomic analysis revealed significant downregulation of sarcomere-related genes and calcium-handling genes, with enrichment in pathways associated with cardiomyopathy and calcium signaling. Furthermore, FHOD3 deficiency triggered the phosphorylation of CaMKII (Thr286), a key regulator of cardiac hypertrophy and remodeling, contributing to the progression of heart failure. Treatment with the myosin activator Omecamtiv mecarbil (OM) partially restored contractility without affecting calcium handling, highlighting its potential as a therapeutic strategy.
Stem Cell Research & Therapy•2025•DOI: 10.1186/s13287-025-04148-1
Background The long-term effects and outcomes of human mesenchymal stem cell (MSC) therapy in patients with severe coronavirus disease 2019 (COVID-19) remain poorly understood. This study aimed to evaluate the extended safety and efficacy of MSC treatment in severe patients with COVID-19 who participated in our earlier randomized, double-blind, placebo-controlled clinical trial, with follow-up conducted over 3 years. Methods One hundred patients with severe COVID-19 were randomized to receive either an MSC infusion (n=65, 4×10^7 cells/dose, on days 0, 3, and 6) or a placebo, with both groups receiving the standard of care. At 36 months post-MSC therapy, patients were followed up to long-term safety and efficacy, particularly the effects of MSC therapy on persistent COVID-19 symptoms. Evaluated outcomes included lung imaging results, 6-min walking distance (6-MWD), pulmonary function test results, quality of life scores based on the Short Form-36 (SF-36) health survey, Long COVID symptoms, new-onset comorbidities, tumor marker levels, and rates of COVID-19 reinfection. Results Three years post-treatment, 46.94% (23/49) of patients in the MSC group and 34.48% (10/29) in the placebo group showed normal findings on computed tomography (CT) images (odds ratio [OR]=1.68, 95% confidence interval [CI]: 0.65–4.34). The general health (GH) score from the SF-36 was higher in the MSC group (67.0) compared to the placebo group (50.0), with a difference of 12.86 (95% CI: 1.44–24.28). Both groups showed similar results for total lung severity scores (TSS), 6-MWD, pulmonary function tests, and Long COVID symptoms. No significant differences between groups were observed in new-onset complications (including tumorigenesis) or tumor marker levels. After adjusting for China’s dynamic zero-COVID-19 strategy, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) reinfection rates were 53.06% (26/49) in the MSC group and 67.86% (19/28) in the placebo group (OR=0.54, 95% CI: 0.20–1.41). Conclusions These findings support the long-term safety of MSC therapy in patients with severe COVID-19 over 3 years. MSC treatment may offer potential benefits for lung recovery and improved quality of life in patients experiencing Long COVID symptoms. Trial registration: ClinicalTrials.gov, NCT04288102. Registered 28 February 2020, https://clinicaltrials.gov/study/NCT04288102.
Stem Cell Research & Therapy•2025•DOI: 10.1186/s13287-025-04213-9
Background Spaceflight and microgravity environments have been shown to cause significant health impairments, including bone loss, immune dysfunction, and hematopoietic disorders. Hematopoietic stem cells (HSCs), as progenitors of the hematopoietic system, are critical for the continuous renewal and regulation of immune cells. Therefore, elucidating the regulatory mechanisms governing HSC fate and differentiation in microgravity environments is of paramount importance. Methods In this study, hindlimb unloading (HU) was employed in mice to simulate microgravity conditions. After 28 days of HU, cells were isolated for analysis. Flow cytometry and colony-forming assays were utilized to assess changes in HSC proliferation and differentiation. Additionally, transcriptomic and untargeted metabolomic sequencing were performed to elucidate alterations in the metabolic pathways of the bone marrow microenvironment and their molecular regulatory effects on HSCs fate. Results Our findings revealed that 28 days of HU impaired hematopoietic function, leading to multi-organ damage and hematological disorders. The simulated microgravity environment significantly increased the HSCs population in the bone marrow, particularly within the long-term and short-term subtypes, while severely compromising the differentiation capacity of hematopoietic stem/progenitor cells. Transcriptomic analysis of HSCs, combined with metabolomic profiling of bone marrow supernatants, identified 1,631 differentially expressed genes and 58 metabolites with altered abundance. Gene set enrichment analysis indicated that HU suppressed key pathways, including hematopoietic cell lineage and MAPK signaling. Furthermore, integrated analyses revealed that metabolites affected by HU, particularly hypoxanthine enriched in the purine metabolism pathway, were closely associated with hematopoietic cell lineage and MAPK signaling pathways. Molecular docking simulations and in vitro experiments confirmed that hypoxanthine interacts directly with core molecules within these pathways, influencing their expression. Conclusions These findings demonstrate that hypoxanthine in the bone marrow supernatant acts as a signaling mediator under microgravity, influencing HSCs fate by modulating hematopoietic cell lineage and MAPK signaling pathways. This study offers novel insights into the impact of microgravity on HSC fate and gene expression, underscoring the pivotal role of bone marrow microenvironmental metabolic changes in regulating key signaling pathways that determine hematopoietic destiny.
Stem Cell Research & Therapy•2025•DOI: 10.1186/s13287-025-04173-0
Background Cavernous nerve injury-induced erectile dysfunction (CNI-ED) is a common complication after radical prostatectomy. Conventional treatment approaches have had little success in treating the severe cavernous fibrosis which is a consequence of CNI-ED. Methods Pre-treatment of adipose-derived stem cells with melatonin allows for the extraction of active exosomes (MT-hASC-EVs) from the conditioned medium. The therapeutic effects of MT-hASC-EVs were assessed in a rat model of CNI-ED, and the anti-fibrotic properties were evaluated. MicroRNA sequencing was used to identify specific microRNAs highly expressed in MT-hASC-EVs, and differential microRNAs were screened for regulatory pathways through target gene enrichment analysis. Finally, the conclusions from bioinformatics analysis were validated through in vitro experiments. Results Intracavernous injection of MT-hASC-EVs significantly restored erectile function and reduced the extent of corpus cavernosum fibrosis in the CNI-ED rat model. MT-hASC-EVs promoted the proliferation and anti-apoptotic effects of corpus cavernosum smooth muscle cells (CCSMCs) in vitro. Mechanistically, MT-hASC-EVs inhibit fibrosis by delivering miR-145-5p, which targets TGF-β2/Smad3 axis. Conclusions MT-hASCs-EVs can inhibit cavernous fibrosis and improve erectile function in a rat model of CNI-ED by targeting the miR-145-5p/TGF-β/Smad axis.
Stem Cell Research & Therapy•2025•DOI: 10.1186/s13287-025-04322-5
Background Nano-zinc oxide (nZnO) has attracted significant attention in bone tissue engineering due to its antibacterial properties, anti-inflammatory effects, biocompatibility, and chemical stability. Although numerous studies have demonstrated the enhancement of osteogenic differentiation by nZnO-modified tissue engineering materials, the underlying mechanisms remain poorly characterized. Methods This study aimed to identify the molecular mechanisms how nZnO promoted osteogenic differentiation and bone regeneration using transcriptome analysis, drug intervention, and shRNA knockdown techniques, etc. First, the study evaluated the in vivo effects of gelatin methacryloyl (GelMA) containing nZnO on bone regeneration using a mouse calvarial defect model. The impact of nZnO exposure on the osteogenic differentiation of mesenchymal stem cells (MSCs) was then assessed. The combined treatment of nZnO and MSCs in GelMA for bone regeneration was assessed in the mouse calvarial defect model thereafter. Results nZnO induced osteoblastic differentiation to promote bone regeneration. nZnO activated the AMP-dependent protein kinase (AMPK)-ULK1 signals to stimulate autophagosomes formation and facilitate autophagy flow, which was the essential pathway to induce osteogenic differentiation. The combined treatment of MSCs and nZnO significantly enhanced bone regeneration in calvarial defect mice. Conversely, AMPK inhibitor Compound C (C.C) reversed the effects on autophagy flow and osteogenic potentiality induced by nZnO. Conclusions These results highlight that nZnO can regulate bone regeneration by activating autophagy through the AMPK/ULK1 signaling pathway, which may provide a novel therapeutic strategy for addressing bone defects using nZnO.
Stem Cell Research & Therapy•2025•DOI: 10.1186/s13287-025-04401-7
Background: Multiple sclerosis (MS) is a progressive autoimmune disease characterized by massive inflammatory infiltration, demyelination, and subsequent axonal injury and neuronal damage in the central nervous system (CNS). The etiology of MS remains unclear and there is not yet a definitive therapeutic schedule for the disease. Bone marrow mesenchymal stem cells (BMSCs), exhibiting neuroimmune-modulatory functions to alleviate various autoimmune diseases, show great potential in the treatment of MS. However, the instability of BMSCs-mediated immunosuppression in vivo has limited their application. MiR181-a, a positive regulator of immune balance, which has a preference for T cells and B cells differentiation, but degrade rapidly upon entering systemic circulation due to their unstable molecular structure. Methods: We propose a synergistic therapy approach that combines the penetrative targeting capability of BMSCs with the immuno-modulatory effects of miR181-a by overexpressing miR181-a to BMSCs through lentivirus packaging system. With this strategy, on the basis of the establishment of the experimental autoimmune encephalomyelitis (EAE) model, miR181-a overexpressing BMSCs (miR181a-BMSCs) would have a stronger immuno-modulatory treatment benefit, in terms of attenuating MS development. Results: Indicate that this method prolongs the modulatory effects of BMSCs and resulted in significantly enhancements of the proliferation of regulatory B cells (Bregs), regulatory T cells (Tregs) and the inhibition of Th17 cells compared to the traditional BMSCs group. Moreover, 10-fold miRNA’s concentration in the exosome of miR181a-BMSCs, leading to an increased duration of miRNAs to exert their biological effects. By immunotherapy and synergistic treatment, the effectiveness of the treatment is significantly enhanced, showing consistent results in different groups of the animal model.
Stem Cell Research & Therapy•2025•DOI: 10.1186/s13287-025-04229-1
Background Despite numerous studies addressing the molecular mechanisms by which pluripotent stem cells (PSCs) maintain self-renewal and pluripotency under normal culture conditions, the fundamental question of how PSCs manage to survive stressful conditions remains largely unresolved. Post-transcriptional/translational regulation emerges to be vital for PSCs, but how PSCs coordinate and balance their survival and differentiation at translational level under extrinsic and intrinsic stress conditions is unclear. Methods The high-throughput sequencing of cross-linking immunoprecipitation cDNA library (HITS-CLIP) was employed to decipher the genome-wide OCT4-RNA interactome in human PSCs, a combined RNC-seq/RNA-seq analysis to assess the role of OCT4 in translational regulation of hypoxic PSCs, and an OCT4-protein interactome to search for OCT4 binding partners that regulate cap-independent translation initiation. By taking the Heterozygous Knocking In N-terminal Tags (HKINT) approach that specifically disrupts the 5'-UTR secondary structure and tagging its protein product of the mRNA from one allele while leaving that from the other allele intact, we examined the effect of disrupting the OCT4/5'-UTR interaction on translation of AKT1 mRNA. Results We revealed OCT4 as a bona fide RNA-binding protein (RBP) in human PSCs that bound to the 5'-UTR, 3'-UTR and CDS regions of mRNAs. Multiple known proteins participating in IRES-mediated translation initiation were detected in the OCT4-protein interactome, and a combined RNC-seq/RNA-seq analysis further confirmed a crucial role of OCT4 in translational regulation of PSCs in response to hypoxic stress. Remarkably, OCT4 bound to the GC-rich elements in the 5'-UTR of AKT1 and multiple PI3K/AKT-pathway-gene mRNAs, and promoted their translation initiation via IRES-mediated pathways under stress conditions. Specifically disrupting the AKT1 mRNA 5'-UTR structure and the OCT4/5'-UTR interaction by the HKINT approach significantly reduced the translation level of AKT1 that led to a higher susceptibility of PSCs to oxidative stress-induced apoptotic death and prioritized differentiation toward ectoderm and endoderm. Conclusions Our results reveal OCT4 as an anti-stress RBP for translational regulation that critically coordinates the survival and differentiation of PSCs in response to various stressors.
Stem Cell Research & Therapy•2025•DOI: 10.1186/s13287-025-04751-2
Background Sepsis survivors frequently develop compromised humoral immunity, manifesting as prolonged immunosuppression that increases susceptibility to secondary infections with high associated mortality. While mesenchymal stromal cells (MSCs) have demonstrated efficacy in mitigating initial inflammatory responses and improving early survival rates in murine cecal ligation and puncture (CLP) models, they show limited effectiveness against subsequent secondary infections. Given that lymphopenia represents a primary driver of this infection vulnerability, we sought to investigate whether spleen-targeted MSC therapy could prevent lymphocyte depletion and enhance protection against secondary infections. Methods In this study, we developed CXCR5-overexpressing MSCs (MSCCXCR5) to enhance their targeted migration toward splenic CXCL13. Using a CLP mouse model followed by secondary Staphylococcus aureus (S. aureus) infection, we systematically evaluated the therapeutic potential of MSCCXCR5 in combating post-sepsis immunosuppression. Results MSCCXCR5 demonstrated enhanced splenic migration, specifically homing to CXCL13-rich B-cell zones, and conferred significant protection against both S. aureus secondary infection and sepsis-induced lymphopenia. Mechanistic investigations revealed that MSCCXCR5 treatment preserved follicular and germinal center B-cell populations, maintaining splenic architecture and potentiating antigen-specific B-cell responses to CLP challenge. These effects collectively contributed to markedly improved survival outcomes in CLP mice following secondary infection. Conclusion Our findings demonstrate that MSCCXCR5 confers dual-phase protection in sepsis, improving survival during both the initial hyperinflammatory phase and subsequent immunosuppressive period. This comprehensive
Stem Cell Research & Therapy•2025•DOI: 10.1186/s13287-025-04241-5
Background Chronic diabetic wounds pose a significant clinical challenge due to the limited efficacy of current treatments. This study aimed to investigate the role and potential mechanisms of adipose-derived mesenchymal stem cells (ADSCs) overexpressing acidic fibroblast growth factor (aFGF) in diabetic wound healing in a rat model. Methods ADSCs were genetically modified to achieve stable overexpression of aFGF. Varying doses of aFGF-ADSCs (1×10⁶, 2×10⁶, 3×10⁶, 4×10⁶) were injected into the muscular tissue surrounding diabetic rat wounds. We assessed aFGF expression and its impact on various stages of wound healing, including angiogenesis, inflammatory response, epithelialization, and collagen deposition. Transcriptomic sequencing was performed to explore the underlying mechanisms driving enhanced wound healing. Results Lentiviral transduction successfully induced stable aFGF overexpression in ADSCs. In vivo experiments revealed that varying doses of aFGF-ADSCs markedly enhanced wound healing in diabetic rats in a dose-dependent manner. The dose of 3×10⁶ aFGF-ADSCs demonstrated the most significant effect. In the 3×10⁶ aFGF-ADSCs group, expression levels of aFGF, CD31, and CD163 were significantly higher than in other groups (p < 0.05), while CD86 expression was significantly lower (p < 0.05). Conclusion Single doses of aFGF-ADSCs comprehensively improved various aspects of wound repair in diabetic rats, offering a potential new approach for treating chronic diabetic wounds. The mechanism of action involves promoting angiogenesis, modulating inflammatory responses, accelerating epithelialization, and optimizing collagen deposition.
Stem Cell Research & Therapy•2024•DOI: 10.1186/s13287-024-03984-x
This correction article addresses an inadvertent error in the original publication. In Fig. 5M of the original article, the image of the fourth lane (LEF-1) of the second picture (Doxorubicin-induced GCSCs) was inadvertently replaced with an incorrect version during the upload process. The authors wish to note a correction to the aforementioned picture via the corrected picture ahead in this Correction article. The authors deeply regret that this error occurred and sincerely apologize for any inconvenience.
Stem Cell Research & Therapy•2024•DOI: 10.1186/s13287-024-03917-8
Background Intermediate cells are present in the early stages of human prostate development and adenocarcinoma. While primary cells isolated from benign human prostate tissues or tumors exhibit an intermediate phenotype in vitro, they cannot form tumors in vivo unless genetically modified. It is unclear about the stem cell properties and tumorigenicity of intermediate cells.
Methods We developed a customized medium to culture primary human intermediate prostate cells, which were transplanted into male immunodeficient NCG mice to examine tumorigenicity in vivo. We treated the cells with different concentrations of dihydrotestosterone (DHT) and enzalutamide in vitro and surgically castrated the mice after cell transplantation in vivo. Immunostaining, qRT-PCR, RNA sequencing, and western blotting were performed to characterize the cells in tissues and 2D and 3D cultures.
Results We found intermediate cells expressing AR+PSA+CK8+CK5+ in the luminal compartment of human prostate adenocarcinoma by immunostaining. We cultured the primary intermediate cells in vitro, which expressed luminal (AR+PSA+CK8+CK18+), basal (CK5+P63+), intermediate (IVL+), and stem cell (CK4+CK13+PSCA+SOX2+) markers. These cells resisted castration in vitro by upregulating the expression of AR, PSA, and proliferation markers KI67 and PCNA. The intermediate cells had high tumorigenicity in vivo, forming tumors in immunodeficient NCG mice in a month without any genetic modification or co-transplantation with embryonic urogenital sinus mesenchyme (UGSM) cells. We named these cells human castration-resistant intermediate prostate cancer stem cells or CriPCSCs and defined the xenograft model as patient primary cell-derived xenograft (PrDX). Human CriPCSCs resisted castration in vitro and
Stem Cell Research & Therapy•2024•DOI: 10.1186/s13287-024-03938-3
Background: Despite the pivotal role of fat grafting in plastic, reconstructive, and aesthetic surgery, inconsistent survival rates of transplanted adipose tissue, primarily due to early ischemic and hypoxic insults, remain a significant challenge. The infusion of healthy mitochondria has emerged as a promising intervention to support tissue recovery from ischemic, hypoxic, and other types of damages across various organ systems. Objectives: This study aims to evaluate the impact of supplementing human adipose tissue grafts with healthy exogenous mitochondria on their volume and mass retention rates when transplanted into the subcutaneous layers of nude mice. This approach seeks to improve and optimize fat grafting techniques. Methods: Human adipose tissues were preconditioned with exogenous mitochondria (10 µg/mL), a combination of exogenous mitochondria and the inhibitor Dyngo-4a, Dyngo-4a alone, or PBS, and then transplanted into the subcutaneous tissue of 24 nude mice. Samples were harvested at 1 and 3 months post-transplantation for analysis of mass and volume retention. The structural morphology and integrity of the adipose tissues were assessed using Hematoxylin and Eosin (H&E) staining. Results: Mitochondrial preconditioning significantly enhanced the retention of mass and volume in fat grafts, demonstrating superior structural morphology and integrity compared to the control group. Conclusions: This study highlights the potential of exogenous mitochondrial augmentation in fat transplantation to significantly improve fat graft survival, thereby optimizing the success of fat grafting procedures.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2026054
The anti-inflammatory properties of the traditional herb Salvia miltiorrhiza Bunge are well-established, yet its precise mechanism of action in ulcerative colitis (UC) remains unclear. Herein, we evaluate the therapeutic potential of four major tanshinones–tanshinone IIA (Tan IIA), miltirone, neocryptotanshinone, and dihydrotanshinone I–in a murine dextran sulfate sodium (DSS)-induced colitis model. Our results show that tanshinones effectively alleviate disease severity, suppress systemic and local inflammation, and restore intestinal barrier integrity. Integrated multi-omics analysis reveals that the therapeutic efficacy originates from a comprehensive reprogramming of the gut microbiota-metabolite axis. Specifically, tanshinones reverse colitis-associated dysbiosis and rectify metabolic disturbances in linoleic acid metabolism, bile acid biosynthesis, and amino acid utilization. Correlation network analysis identifies key functional modules linking beneficial microbes (e.g., Akkermansia) to anti-inflammatory lipid mediators and associating pathobionts (e.g., Desulfovibrio) with disrupted bile acid metabolism. Notably, supplementation with Akkermansia muciniphila synergizes with Tan IIA to amplify barrier restoration and metabolic normalization. Our findings establish that tanshinones ameliorate UC through microbiota-driven metabolic reprogramming, wherein the restructured microbial community actively shapes a therapeutic metabolic output. This work elucidates a metabolite-mediated mechanism of action and positions tanshinones as promising microbiome-targeting therapeutics for inflammatory bowel disease.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2025055
Platelets circulate in an inactive form in the body until they contact with defective areas of endothelial cells or encounter a clotting cascade [1]. Activated platelets release and express bioactive substances and acquire the ability to bind plasma fibrinogen. Abnormal activation of platelets is involved in atherosclerosis and thrombosis [2,3]. When platelets are stimulated and activated, phospholipase A2 is activated at the same time, which then cleaves membrane phospholipids and frees arachidonic acid (AA) [4]. The latter catalyzes thromboxane A2 (TXA2) via thromboxane synthetase [5], which is induced by the cyclooxygenase COX-1 to produce prostaglandins G2 and H2 [6]. TXA2 is highly unstable, with a half-life of only 30 s, and it is rapidly hydrolyzed to relatively stable thromboxane B2 (TXB2), which is then converted in the liver to 11-dehydrothromboxane B2 (11dH-TXB2), which has a longer half-life and is excreted in the urine [7]. Dehydrothromboxane B2 is the final stable metabolite of thromboxane A2, which is derived only from arachidonic acid metabolism and can represent thromboxane A2 level in the body [8]. Specifically, by inhibiting the action of COX-1, the most important enzyme in the process of arachidonic acid metabolism, aspirin inhibits the production of thromboxane A2, that is, the concentration of TXA2 affects the effect of aspirin on platelet aggregation. However, the half-life of TXA2 (including the metabolic intermediate TXB2) is too short to be accurately measured, so the detection of its metabolic end product 11dH-TXB2 can very accurately reflect the sensitivity of the body to aspirin [7]. The concentration of 11dH-TXB2 in the serum correlates well with the concentration of 11dH-TXB2 in the urine, so the determination of 11dH-TXB2 in the urine can more effectively reflect the production of TXA2 in vivo [8]. The 11dH-TXB2 concentration needs to be corrected with the urinary creatinine concentration to rule out the effects of the urine concentration and renal function, so random urine samples can be used for testing [6]. Aspirin can acetylate serine at the key site of cyclooxygenase and thus irreversibly inhibits the activity of COX-1, reduces the synthesis of TXA2, and blocks the production of TXA2 and its induced platelet aggregation. Low-dose aspirin (30–75 mg/day) can effectively inhibit 95% of COX-1 activity [7]. Since the production of TXA2 in serum is largely dependent on platelet COX-1 (a therapeutic target of aspirin), 11dH-TXB2 can be used as a monitor for aspirin-induced platelet inhibition [3]. Creatinine is a metabolic byproduct of muscle metabolism that is primarily excreted via glomerular filtration, and its level is indicative of renal function [9]. The 24-h creatinine clearance can also be used to determine the integrity of the sample or to correct the urine sample concentration with the creatinine ratio [10]. Currently, creatinine detection methods include the Jaffe method, enzymolysis spectrophotometry, HPLC, capillary electrophoresis, capillary zone electrophoresis, gas chromatography tandem mass spectrometry (GC-MS) and liquid chromatography tandem mass spectrometry (LC-MS/MS) [11]. At present, there is no method for the simultaneous detection of 11dH-TXB2 and creatinine. When both analytes are needed, separate tests must be performed, increasing the workload and sample volume requirements. Thus, developing a method that enables the concurrent quantification of 11dH-TXB2 and creatinine in a single assay remains a critical challenge. The aim of this study was to provide a method for the simultaneous detection of 11dH-TXB2 and creatinine and to alleviate the problem that 11dH-TXB2 and creatinine cannot be simultaneously detected. By developing a standardized quantitative approach for measuring 11dHTXB2 and creatinine in human urine, this study aims to provide reliable concentration data, thereby facilitating further clinical research and methodology optimization.
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.2025056
Triptonide (TN) is a small-molecule compound initially derived from Tripterygium wilfordii Hook. f used in traditional Chinese medicine. However, its potential antitumor mechanisms are still far from adequately understood. The purpose of this research is to elucidate the antitumor and pharmacological effects of TN on esophageal squamous cell carcinoma (ESCC). Functional assays, such as CCK-8 and colony formation assays, are used to evaluate the effects of TN on KYSE450 and KYSE510 cells. Subsequently, western blot analysis, Hoechst 33258 staining, flow cytometric analysis, autophagic flux detection, and transmission electron microscopy (TEM) are used to determine the effects of TN on apoptosis and autophagy in ESCC cells. Additionally, the autophagy inhibitor 3-methyladenine (3-MA) and the AMPK inhibitor dorsomorphin (Compound C, CC) are administered to explore the molecular mechanisms and crucial pathways in ESCC cells. Our findings provide strong evidence that TN induces autophagy-dependent apoptosis by targeting the AMPK-mTOR-ULK1 axis in ESCC cells. Collectively, this study sheds light on the anticancer mechanisms of TN in esophageal squamous cell carcinoma and suggests that TN is a promising candidate for the antitumor phytomedicine.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2025102
Influenza A viruses (IAVs) constitute a major threat to human and animal health. Currently, M2 ion-channel inhibitors, neuraminidase (NA) inhibitors, RNA polymerase inhibitors, and cap-dependent endonuclease inhibitors have been applied clinically as therapeutics against IAVs. However, IAVs possess adaptive mutations to these inhibitors, especially M2 ion channel and NA inhibitors. Thus, novel antiviral agents should be developed. In the present study, we screen approximately 5500 compounds and identify an IAV inhibitor, nanchangmycin, which possesses a robust antiviral activity both in vitro and in vivo. In addition, it exhibits broad-spectrum antiviral activity for additional virus infections, including pseudorabies virus, herpes virus, porcine epidemic diarrhea virus, porcine reproductive and respiratory syndrome virus. Most importantly, it has antiviral activity against oseltamivir-resistant strains in sub-μM ranges and promotes the survival of MDCK cells infected with the oseltamivir-resistant influenza A virus strain. Further studies reveal that it blocks the nuclei migration of viral nuclear proteins (NPs), resulting in NP accumulation in the cytoplasm, particularly within perinuclear endosomes. Also, it inhibits IAVs by blocking endosomal acidification. Overall, nanchangmycin has the potential to be developed as an anti-influenza agent.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2025042
This study elucidates the mechanisms by which cancer-associated fibroblast (CAF)-derived platelet-derived growth factor C (PDGFC) promotes the progression of lung adenocarcinoma (LUAD) and explores the impact of PDGFC on immune regulation within the tumor microenvironment (TME). Our results show that there is higher expression of PDGFC in CAFs than in nontumor tissue fibroblasts (NFs) and that higher expression of PDGFC is correlated with poor prognosis in LUAD patients. Furthermore, CAF-derived PDGFC promotes epithelial-mesenchymal transition (EMT) in cancer cells as well as matrix metalloproteinase 2 (MMP2) expression through the PDGF receptor A (PDGFRA)-mitogen-activated protein kinase/extracellular signal-regulated kinase (MAPK/ERK) pathway. Moreover, our study demonstrates that CAF-derived PDGFC is essential for the activation and infiltration of fibroblasts in the TME, as well as the inflammatory infiltration of different immune cell types and the immunosuppressive conditions within the TME. In particular, PDGFC induces increased PDGFRA expression in both tumor cells and fibroblasts, which can lead to reciprocally positive feedback to accelerate malignant tumor progression. This discovery provides a novel TME-targeted strategy for LUAD treatment.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2025043
Angiogenesis is an important factor influencing the development of solid tumors, and vascular endothelial growth factor receptor-2 (VEGFR2) is a central regulator of angiogenesis. Antibodies and inhibitors against VEGFR2 have been widely used in various malignancies. However, the regulatory mechanism of VEGFR2 has not been fully clarified. Here, we show that D-mannose can significantly inhibit angiogenesis and tumor growth by degrading VEGFR2. Specifically, D-mannose inactivates GSK3β by promoting the phosphorylation of GSK3β at Ser9, enhances the nuclear translocation of TFE3, and promotes lysosomal biogenesis, thereby increasing the lysosome-mediated degradation of VEGFR2. Thus, D-mannose significantly inhibits the proliferation, migration, and capillary formation of human umbilical vein endothelial cells (HUVECs) in vitro. Oral administration of D-mannose dramatically inhibits angiogenesis and tumor growth in mice. Our findings reveal a previously unrecognized anti-tumor mechanism of D-mannose by destabilizing VEGFR2 and provide a new strategy for the clinical treatment of colorectal cancer (CRC).
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2025129
This is a corrigendum to the article 'Magnolol promotes the autophagy of esophageal carcinoma cells by upregulating HACE1 gene expression' published in Acta Biochimica et Biophysica Sinica 2024, 56(7): 1044–1054. In the original publication, the corresponding author's email address was personal. To comply with the institution's publishing policy, it has been changed from '[email protected]' to the institutional address '[email protected]'. The authors apologize for any confusion it may have caused.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2025025
The circadian rhythm is a phenomenon in which physiological, behavioral, and biochemical processes within an organism naturally fluctuate over a period of approximately 24 hours. This phenomenon is ubiquitous in living organisms. Disruption of circadian rhythms in mammals leads to different diseases, such as cancer, and neurodegenerative and metabolic disorders. In specific tissues, numerous genes have been found to have circadian oscillations, suggesting a broad role for rhythm genes in the regulation of gene expression. This review systematically summarizes the role of cryptochromes (CRYs) in the initiation and progression of different types of cancer and discusses the relationships between clock genes and the tumor microenvironment (TME), as well as clock-based therapeutic strategies.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2025142
Dolutegravir (DTG) disrupts mouse embryonic development in a dose-dependent manner, culminating in neural-tube defects (NTDs). Using whole embryo culture (WEC), mouse embryos at embryonic day 8.5 (E8.5) are cultured for 24–48 h with 8, 10, or 12 μM DTG. The results reveal that higher DTG concentrations dose-dependently disrupt yolk sac development and markedly increase the frequency of NTDs. In vivo NTD models are generated by intraperitoneally injecting DTG at a dose of 7.5 mg/kg, and the resulting embryos exhibit disrupted yolk sac blood circulation, embryonic growth restriction, and malformations. Mechanistic studies suggest that DTG contributes to NTDs by inducing apoptosis: DTG exposure activates the Nrf2-SOD1/CAT antioxidant axis, yet it culminates in increased apoptosis and suppressed proliferation, ultimately impairing yolksac vasculogenesis and neuralepithelial closure, thereby producing NTDs. This study provides new evidence for assessing the potential risk of DTG in embryonic development and highlights the need to re-evaluate its clinical safety in future applications.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2025027
Currently, chemotherapy remains the primary treatment for acute myeloid leukemia (AML). Drug resistance in AML cells is a critical factor contributing to the failure of chemotherapy remission and subsequent relapse. Iron overload frequently occurs in AML patients because of hematopoietic suppression or supportive blood transfusion therapy. Previous studies have indicated that iron overload may promote the progression of AML; however, the underlying mechanisms remain unclear. Our results demonstrate that, compared with TP53-wild-type AML cells, TP53-mutant AML cells exhibit increased resistance to cytarabine-induced cytotoxicity. Moreover, reducing TP53 expression in wild-type AML cells diminishes their sensitivity to cytarabine. The TP53 signaling pathway is essential for mediating cytarabine-induced apoptosis in AML cells. In this study, an iron overload model in AML cells via the use of ferric citrate is constructed. Our data indicate that iron overload can suppress the TP53/BCL2/BAX signaling pathway, counteracting cytarabine-induced apoptosis. In TP53 wild-type AML cells, TFR1 participates in iron-mediated resistance to cytarabine by regulating the entry of iron into the cells. These findings provide a foundation for further exploration of the molecular mechanisms involved in AML resistance to cytarabine.
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.2025197
Acute myeloid leukemia (AML) is a clinically aggressive hematologic malignancy characterized by high relapse rates and treatment resistance, highlighting the need for novel biomarkers to improve clinical outcomes. In this study, we explore the roles of nuclear receptor-interacting protein 1 (NRIP1) in AML, focusing on its associations with tumor progression and immune infiltration. Analysis of public AML gene expression datasets reveals that NRIP1 expression is significantly increased in AML patients. Those with high NRIP1 expression have markedly shorter overall survival than those with low expression. Furthermore, NRIP1 expression is significantly associated with the infiltration of diverse immune cells, including B cells, dendritic cells, T cells, mast cells, eosinophils, and T helper cells, suggesting that NRIP1 may be a regulator of immune cell infiltration. Functional enrichment analysis indicates that NRIP1 and its interacting partners are involved in tumorigenesis, immune microenvironment remodeling, and metabolic reprogramming. Survival analysis confirms the prognostic value of NRIP1. Importantly, functional validation in AML cell lines confirms that NRIP1 knockdown suppresses proliferation and induces apoptosis. Our study identifies NRIP1 as a multifaceted regulator that promotes AML by driving tumor progression, regulating immune cell infiltration, and modulating ferroptosis, highlighting its role as a novel prognostic biomarker.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2025120
The phosphoenolpyruvate-dependent sugar phosphotransferase system (PTS) is a central pathway for carbohydrate transport in bacteria and plays a critical role in nutrient acquisition, metabolism, and virulence. In Vibrio cholerae, the glucose-specific EIIC transporter is a key component of the PTS system, mediating the transport of sugars into the bacterial cell, coupled with phosphorylation during translocation. Here, we present the 3.68 Å cryo-electron microscopy (cryo-EM) structure of the dimeric EIIC transporter from Vibrio cholerae in its inward-facing, substrate-free conformation. The structure reveals a detailed arrangement of the scaffold and transport domains, stabilized by extensive inter- and intraprotomer interactions. Comparative analysis with substrate-bound inward-facing structures of EIIC from E. coli highlights conformational changes, providing insights into substrate release and the structural transitions required for alternating access. Notably, the observed substrate-free inward-facing conformation features a larger substrate-binding pocket, which is consistent with a state poised for glucose release into the cytoplasm. The formation of a unique intraprotomer disulfide bond between residues C240 and C254 stabilizes the interface between the scaffold and transport domains, potentially regulating transporter dynamics. These findings elucidate the structural basis for substrate release in the PTS system and underscore the dynamic nature of EIIC-mediated sugar transport. Our study enhances the understanding of PTS system function in Vibrio cholerae and highlights the EIIC transporter as a promising target for antimicrobial drug development. Disruption of sugar transport in this essential pathway could impair bacterial growth and virulence, suggesting a novel therapeutic strategy against cholera. These results provide a foundation for future investigations into the structural and functional dynamics of bacterial sugar transporters.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2024200
This is a corrigendum to the original article published in Acta Biochim Biophys Sin 2020, 52(2): 116–124. The authors correct an error in Figure 4 of the original manuscript. The correct figure is provided, and the authors apologize for the error.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2025196
Human embryonic kidney (HEK) 293 cells are widely used for recombinant protein production because of their efficient posttranslational modification capabilities. However, their large-scale culture is often limited by metabolic stress and early apoptosis, leading to insufficient protein yields. In this study, we aim to increase protein expression through the coordinated modulation of metabolic and apoptotic pathways. Using CRISPR/Cas9 technology, we target and knockout the genes of ornithine decarboxylase antizyme 1 (OAZ1), which regulates polyamine metabolism, and caspase 8-associated protein 2 (CASP8AP2), an apoptosis-related protein. We successfully construct an OAZ1/CASP8AP2 double-knockout HEK293 cell line. Following transfection with the knockout vector and screening of single-cell clones, multiple levels of validation confirm the successful gene knockout. The results show that the double-knockout cells exhibit significantly reduced apoptosis rates. Furthermore, the production of recombinant secreted alkaline phosphatase (SEAP) and vitronectin (VN) increases by 2.1 folds and 2.9 folds, respectively, compared with those in wild-type cells. Metabolic profiling reveals that the cell cycle is arrested in the G1/G0 phase, accompanied by increased specific consumption and production rates of key metabolites. This study demonstrates that concurrent inhibition of apoptosis and optimization of metabolism effectively enhances recombinant protein production in HEK293 cells, suggesting a novel strategy for improving HEK293 cell-based expression.
Acta Biochimica et Biophysica Sinica•2024•DOI: 10.3724/abbs.2024084
Osteosarcoma (OS) is a primary bone cancer mostly found in adolescents and elderly individuals. The treatment of OS is still largely dependent on traditional chemotherapy. However, the high incidence of drug resistance remains one of the greatest impediments to limiting improvements in OS treatment. Recent findings have indicated that the transcription factor FOXM1 plays an important role in various cancer-related events, especially drug resistance. However, the possible role of FOXM1 in the resistance of OS to methotrexate (MTX) remains to be explored. Here, we find that FOXM1, which confers resistance to MTX, is highly expressed in OS tissues and MTX-resistant cells. FOXM1 overexpression promotes MTX resistance by enhancing autophagy in an HMMR/ATG7-dependent manner. Importantly, silencing of FOXM1 or inhibiting autophagy reverses drug resistance. These findings demonstrate a new mechanism for FOXM1-induced MTX resistance and provide a promising target for improving OS chemotherapy outcomes.
Acta Biochimica et Biophysica Sinica•2024•DOI: 10.3724/abbs.2024049
Chronic renal failure (CRF) is a severe syndrome affecting the urinary system for which there are no effective therapeutics. In this study, we investigate the effects and mechanisms of aminophylline in preventing CRF development. A rat model of chronic renal failure is established by 5/6 nephrectomy. The levels of serum creatinine (SCR), urinary protein (UPR), and blood urea nitrogen (BUN) are detected by ELISA. Histological evaluations of renal tissues are performed by H&E, Masson staining, and PAS staining. Functional protein expression is detected by western blot analysis or immunofluorescence microscopy. Glomerular cell apoptosis is determined using the TUNEL method. Results show that Aminophylline significantly reduces the levels of SCR, UPR, and BUN in the CRF model rats. Histological analyses show that aminophylline effectively alleviates renal tissue injuries in CRF rats. The protein expression levels of nephrin, podocin, SIRT1, p-AMPK, and p-ULK1 are greatly increased, while p-mTOR protein expression is markedly decreased by aminophylline treatment. Additionally, the protein level of LC3B in CRF rats is significantly increased by aminophylline. Moreover, aminophylline alleviates apoptosis in the glomerular tissues of CRF rats. Furthermore, resveratrol promotes SIRT1, p-AMPK, and p-ULK1 protein expressions and reduces p-mTOR and LC3B protein expressions in CRF rats. Selisistat (a SIRT1 inhibitor) mitigates the changes in SIRT1, p-AMPK, p-ULK1, p-mTOR, and LC3B expressions induced by aminophylline. Finally, RAPA alleviates renal injury and apoptosis in CRF rats, and 3-MA eliminates the aminophylline-induced inhibition of renal injury and apoptosis in CRF rats. Aminophylline suppresses chronic renal failure progression by modulating the SIRT1/AMPK/mTOR-mediated autophagy process.
Acta Biochimica et Biophysica Sinica•2024•DOI: 10.3724/abbs.2024050
The interaction between bacteria and the host plays a vital role in the initiation and progression of systemic diseases, including gastrointestinal and oral diseases, due to the secretion of various virulence factors from these pathogens. GroEL, a potent virulence factor secreted by multiple oral pathogenic bacteria, is implicated in the damage of gingival epithelium, periodontal ligament, alveolar bone and other peripheral tissues. However, the underlying biomechanism is still largely unknown. In the present study, we verify that GroEL can trigger the activation of NLRP3 inflammasome and its downstream effector molecules, IL-1β and IL-18, in human periodontal ligament stem cells (hPDLSCs) and resultantly induce high activation of gelatinases (MMP-2 and MMP-9) to promote the degradation of extracellular matrix (ECM). GroEL-mediated activation of the NLRP3 inflammasome requires the participation of Toll-like receptors (TLR2 and TLR4). High upregulation of TLR2 and TLR4 induces the enhancement of NF-κB (p-p65) signaling and promotes its nuclear accumulation, thus activating the NLRP3 inflammasome. These results are verified in a rat model with direct injection of GroEL. Collectively, this study provides insight into the role of virulence factors in bacteria-induced host immune response and may also provide a new clue for the prevention of periodontitis.
Acta Biochimica et Biophysica Sinica•2024•DOI: 10.3724/abbs.2024100
Tuberculosis (TB), caused by Mycobacterium tuberculosis (M.tb), remains one of the leading causes of fatal infectious diseases worldwide. The only licensed vaccine, Mycobacterium bovis Bacillus Calmette-Guérin (BCG), has variable efficacy against TB in adults. Insufficiency of immune cell function diminishes the protective effects of the BCG vaccine. It is critical to clarify the mechanism underlying the antimycobacterial immune response during BCG vaccination. Macrophage mannose receptor (MR) is important for enhancing the uptake and processing of glycoconjugated antigens from pathogens for presentation to T cells, but the roles of macrophage MR in the BCG-induced immune response against M.tb are not yet clear. Here, we discover that macrophage MR deficiency impairs the antimycobacterial immune response in BCG-vaccinated mice. Mechanistically, macrophage MR triggers JAK-STAT1 signaling, which promotes antigen presentation via upregulated MHC-II and induces IL-12 production by macrophages, contributing to CD4+ T cell activation and IFN-γ production. MR deficiency in macrophages reduces the vaccine efficacy of BCG and increases susceptibility to M.tb H37Ra challenge in mice. Our results suggest that MR is critical for macrophage antigen presentation and the antimycobacterial immune response to BCG vaccination and offer valuable guidance for the preventive strategy of BCG immunization.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2025019
Cancer cells evade immune detection through checkpoint molecules like PD-L1 and PD-L2 which suppress T-cell activation. While PD-L1 is well-studied, the role of PD-L2 remains unclear. Pyruvate kinase M2 (PKM2), a metabolic enzyme, influences immune checkpoint regulation, but its role in PD-L1 and PD-L2 modulation is not well defined. Here, we investigate the role of pyruvate kinase M2 (PKM2) in modulating the immune checkpoint molecules PD-L1 and PD-L2 via GATA3 in cancer cells, with insights from both human and mouse models. We find that PKM2 enhances PD-L1 expression while inhibiting PD-L2, a dual regulatory mechanism that facilitates immune evasion. Knockdown and overexpression experiments revealed GATA3 as a key mediator. PKM2 knockout reduced GATA3 level, leading to decreased PD-L1 and increased PD-L2 expression. Chromatin immunoprecipitation (ChIP)-qPCR demonstrates that GATA3 functions as a direct transcription factor capable of binding to the promoters of PD-L1 and PD-L2. In silico analyses of 81 esophageal squamous cell carcinoma (ESCC) cases from the TCGA database demonstrate that PKM2 mRNA is unrelated to PD-L1 and PD-L2 expression but is negatively correlated with CD8+ T-cell infiltration in ESCC. To further validate these findings, we establish a xenograft model using immune-competent C57/BL6N mice, where knockdown of PKM2 results in significant downregulation of both PD-L1 and PD-L2 expression. Collectively, these findings underscore the divergent roles of PKM2 in regulating immune checkpoint expression in human and mouse cancer models and suggest that targeting the PKM2-GATA3 axis could enhance cancer immunotherapy by fine-tuning PD-L1 and PD-L2 levels.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2025052
Platinum drugs are widely used in lung cancer chemotherapy, but the immune characteristics of different individuals have different effects on the sensitivity and side effects of platinum drugs. In this study, we use 731 kinds of immune cell traits of 3757 healthy individuals and 429 patients with non-small cell lung cancer (NSCLC) in Xiangya Hospital of Central South University to conduct a Mendel randomized analysis in order to find out the causal relationship between some immune cell traits and the efficacy and adverse reactions of platinum drugs. We find that CD19 on CD24+CD27+ B cell (OR = 0.598, P = 0.004) is the most significant immune cell trait as the protective factor of efficacy. HLA-DR+CD8+ T cell % lymphocyte (OR = 0.427, P = 7.55 × 10–4) and HLA-DR+CD8+ T cell % T cell (OR = 0.471, P = 0.003) are the protective factors of liver injury. CD39 on CD39+ secreting CD4+ regulatory T cell (OR = 28.729, P = 0.009) and CD3 on CD39+ resting CD4 regulatory T cell (OR = 3.024, P = 0.009) are the risk factors of renal injury. Meanwhile, B cell-related traits mainly affect gastrointestinal upset and cutaneous toxicity, while T cell-related traits mainly affect other outcome variables. These findings may promote our understanding of the relationship between the efficacy and adverse reactions of platinum drugs and the immune system, and promote future development of biomarkers for predicting the efficacy and adverse reactions of platinum drugs.
Acta Biochimica et Biophysica Sinica•2024•DOI: 10.3724/abbs.2024044
Esophagus cancer (EC) is one of the most aggressive malignant digestive system tumors and has a high clinical incidence worldwide. Magnolol, a natural compound, has anticancer effects on many cancers, including esophageal carcinoma, but the underlying mechanism has not been fully elucidated. Here, we first find that magnolol inhibits the proliferation of esophageal carcinoma cells and enhances their autophagy activity in a dose- and time-dependent manner. This study demonstrates that magnolol increases the protein levels of LC3 II, accompanied by increased HACE1 protein levels in both esophageal carcinoma cells and xenograft tumors. HACE1-knockout (KO) cell lines are generated, and the ablation of HACE1 eliminates the anti-proliferative and autophagy-inducing effects of magnolol on esophageal carcinoma cells. Additionally, our results show that magnolol primarily promotes HACE1 expression at the transcriptional level. Therefore, this study shows that magnolol primarily exerts its antitumor effect by activating HACE1-OPTN axis-mediated autophagy. It can be considered a promising therapeutic drug for esophageal carcinoma.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2024198
Signal regulatory protein α (SIRPα) is recognized as a significant transmembrane protein within the glomeruli that is specifically localized in podocytes, where it plays a role in modulating downstream signaling pathways through phosphorylation. Upon tyrosine phosphorylation of the immunoreceptor tyrosine-based inhibitory motif (ITIM) within SIRPα, protein tyrosine phosphatases are recruited to facilitate the dephosphorylation of downstream signals. Nevertheless, the specific downstream signaling pathways affected by this mechanism have yet to be elucidated. In this study, phosphoproteomic analysis is conducted on podocytes with SIRPα deficiency to identify proteins whose phosphorylation is regulated by SIRPα and the associated signaling pathways in human podocytes. The results reveal significant alterations in biological processes related to cytoskeleton arrangement and cytoskeleton protein binding. Specifically, an increase in FAK tyrosine phosphorylation at Y576 is identified as a potentially crucial signal of the influence of SIRPα on the podocyte cytoskeleton. Our study suggests that SIRPα may facilitate podocyte cytoskeleton rearrangement and migration through the Src/FAK/p38 MAPK signaling pathway. For the first time, we discover increased level of SIRPα, which is strongly linked to urinary protein, in the urine of patients with nephrotic syndrome (NS). Additionally, an increase in urinary FAK level is observed in NS patients, which is positively correlated with both urinary protein level and urinary SIRPα level. These findings suggest that SIRPα and FAK may serve as promising biomarkers for podocytopathies.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2025017
Studies of cell-to-cell activities in the tumor microenvironment (TME) have identified multiple potential targets for oncotherapy. The interplay between tumor cells and neighboring cancer-associated fibroblasts (CAFs) persists in all stages of tumor progression. In this study, we reveal that exosomes from breast cancer cells can be endocytosed into fibroblasts and transform normal fibroblasts (NFs) into CAFs and that the ability of exosomes from highly metastatic breast cancer cells is greater than that of those from poorly metastatic breast cancer cells. Further investigation reveals that exosomes from highly metastatic breast cancer cells contain much more miR-105-5p than those from poorly metastatic breast cells do and that exosomal miR-105-5p facilitates the transformation of NFs to CAFs. A detailed study reveals that RBMY1A1-dependent sorting of miR-105-5p into fibroblasts and subsequent internalization of miR-105-5p promote the transformation of NFs to CAFs by downregulating LATS2 expression and activating NF-κB signaling, which concurrently facilitates the EMT of breast cancer cells. Thus, our results indicate that exosomal miR-105-5p may be a potential target for novel therapeutic strategies to prevent the coevolution of breast cancer cells and CAFs.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2024222
We aim to identify molecular clusters related to O-GlcNAcylation and establish a novel scoring system for predicting prognosis and immunotherapy efficacy in patients with gastric cancer (GC). The transcriptomic and clinical data are obtained from XENA-UCSC and GEO databases. The O-GlcNAcylation-related genes are obtained from the GSEA database. Consensus clustering analysis is employed to identify O-GlcNAcylation-related molecular clusters, and principal component analysis (PCA) is utilized to develop a novel prognostic scoring system for predicting GC outcomes and immunotherapy efficacy. The prognostic accuracy of the scoring system is assessed across five real-world cohorts. The biological function of actin alpha 2, smooth muscle (ACTA2) in GC is determined through experimental verification. Using 34 O-GlcNAcylation-related genes associated with prognosis in GC patients, these individuals are divided into two distinct subgroups characterized by different outcomes, tumor microenvironment profiles, and clinical case characteristics. The DEGs between the two subgroups are subsequently used to further divide the GC patients into two subgroups by consensus cluster analysis. PCA is used to construct a prognostic scoring system, which reveal that patients in the low-score subgroup have a better prognosis and greater benefit from immunotherapy. The accuracy of the scoring system is confirmed through validation in a cohort of patients receiving immunotherapy in the real world. ACTA2 promotes proliferation and inhibits apoptosis in GC cells. These findings suggest that we successfully establish molecular clusters associated with O-GlcNAcylation and develop a scoring system that demonstrates strong performance in predicting the prognosis of patients with GC and the effect of immunotherapy interventions.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2025007
Frailty is a common geriatric disease characterized by accelerated aging and the loss of biological reserves across multiple organs. Approximately 10% of people aged 65 years and older and 25%–50% of people older than 85 years are in a frail state. The increasing institutionalization, hospitalization, and mortality caused by frailty incur massive medical costs and impose a heavy health service burden. For elderly individuals with chronic and/or infectious diseases, such as COVID-19, concomitant frailty can lead to extremely high mortality rates. Moreover, except exercise and nutritional intervention, no effective medicine for treating frailty is available. However, frailty can be prevented, and prefrailty can be reversed. Therefore, effectively screening frailty in elderly individuals is a public health priority. Two main methods for assessing frailty exist: the Fried phenotype and the Rockwood frailty index. The Fried phenotype uses five items, namely, fatigue, weakness, slowness, low physical activity, and weight loss, whereas the Rockwood frailty index is based on the accumulation of age-related deficits. However, these two diagnostic tools are subjective, challenging to use and time-consuming, and are therefore unsuitable for simple, rapid, and extensive screening of frailty in clinical practice. Here, we propose a new strategy to address the above issue. We first harnessed common professional databases to perform inflammatory niche analysis for plasma proteomics from normal aging and frailty patients. We subsequently performed frailty screening and blood sample collection. A total of 852 elderly people were included in the study from January 2018 to August 2018. The assessments included demographic information collection, frailty evaluation, and physical and body composition tests. Blood samples for the determination of inflammatory cytokines were taken from 67 participants. We utilized ELISA to detect the expressions of inflammatory cytokines and chemokines. All blood samples were collected and centrifuged at 4°C and 2000 g for 20 min. The serum was aliquoted and stored properly for ELISA. Inflammatory cytokines in human sera were quantified using corresponding human ELISA kits according to the manufacturer’s protocols. The following markers were measured: IL1A, IL2, IL6, IL8, IL10, IL17, TNFα, IFNγ, GCSF, MCP2, CXCL1, CX3CL1, MMP7, and SOD1. All the statistical analyses were performed with Prism v.6.0. P < 0.05 was considered statistically significant. The receiver operating characteristic (ROC) curve was used to evaluate the performance of all the screening tools. Our inflammatory niche analysis revealed intriguing results when five datasets containing a large number of proteins related to normal aging and inflammation were utilized. The Venn diagrams in Figure 1 show 77 human senescence-associated secretory phenotype genes. Among these genes, 26 are positively correlated with normal aging, whereas 8 are negatively correlated with normal aging. However, neither are positively correlated with frailty, and only two genes are negatively correlated with frailty. Therefore, frailty is obviously distinct from normal aging. Given that most of these differential proteins are inflammatory factors, frailty and normal aging may involve different inflammatory niches. These results suggest that inflammatory factors may be candidates for frailty screening. Our ELISA detection of 15 frailty-related inflammatory factors in the serum of frailty patients screened from 852 volunteers also provided valuable results. The prevalence of frailty was 7.16% (61/852), and that of prefrailty was 41.90% (357/852). The volunteers were 65.22% female and 34.78% male, with a mean age of 70.18 ± 0.73 years. As shown in Table 1, the frail and prefrail groups were significantly older than the robust group (P < 0.001). We observed statistically significant differences in RASM, gait speed, CCI, SARC-F, ADL and MNA scores (P < 0.05), whereas no significant differences were detected with respect to sex, WHR, grip strength or drug count among the three groups. A total of 67 serum samples (nonfrail, n = 20; prefrail, n = 28; and frail, n = 19) were subjected to inflammatory factor screening. No significant difference in the expression of most inflammatory factors was detected.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2024168
Uncoupling protein-2 (UCP2) controls the antioxidant response and redox homeostasis in cancer and is considered a potent molecular target for cancer treatment. However, the specific mechanism of UCP2 inhibition and its role in glioblastoma (GBM) have not yet been elucidated. Here, we attempt to identify a UCP2 inhibitor and study the underlying molecular mechanism in GBM. Bioinformatics analysis and immunohistochemistry are used to validate the high expression of UCP2 in GBM and its prognostic significance. Drug intervention and tumor xenograft experiments are conducted to determine the inhibitory effect of genipin, a UCP2 inhibitor, on UCP2. The mitochondrial membrane potential and key ferroptosis genes are examined to determine the occurrence of ferroptosis. High expression of UCP2 in GBM is associated with poor prognosis, and inhibiting UCP2 can alleviate the malignant behavior of GBM tumors. Genipin can downregulate the expression of GPX4 and upregulate the expression of ACSL4 by inhibiting UCP2, leading to ferroptosis and alleviating the malignant behavior of tumors. In summary, UCP2 is a potential therapeutic target for GBM. Genipin, which targets UCP2, effectively inhibits GBM development by inducing ferroptosis in vivo and in vitro. These findings indicate that genipin treatment based on UCP2 targeting has potential therapeutic applications with a clinical perspective for the treatment of GBM patients.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2024171
Ozone (O3), a prevalent atmospheric pollutant, can induce lung injury. However, the molecular mechanisms of O3-induced acute lung inflammatory injury remain unclear. In this study, we investigate the abnormal changes in and molecular mechanism of mitochondrial homeostasis in alveolar macrophages (AMs) in O3-induced acute lung inflammatory injury mice. Mitochondria and mitochondrial reactive oxygen species (mtROS) are labeled with Mito-Tracker® Deep Red and MitoSOX Red, respectively. Mitochondrial DNA (mtDNA) in AMs from the bronchoalveolar lavage fluid (BALF) is detected via real-time PCR, and the expressions of mitochondrial fusion/fission-related and biogenesis-related proteins in AMs are determined via immunofluorescence staining. Our data show that in O3-induced acute lung inflammatory injury mice, the number of AMs and the protein expression of the NLRP3 inflammasome complex in the lung tissue are increased. In AMs from O3-exposed mice, the number of mitochondria, mtROS, and fission-related protein DRP1 are increased, but the levels of Na+-K+-ATPase, fusion-related protein OPA1, biogenesis-related protein NRF1 and mtDNA are significantly decreased. Compared with that in O3-exposed WT mice, lung inflammation is attenuated, especially the indicators of mitochondrial homeostatic imbalance in AMs, which are alleviated in NLRP3‒/‒ and Caspase-1‒/‒ mice after O3 exposure. These findings indicate that the NLRP3 inflammasome-mediated imbalance in mitochondrial homeostasis in AMs contributes to O3-induced acute lung inflammatory injury. This study may provide a new target for the prevention of lung inflammation induced by O3.
Acta Biochimica et Biophysica Sinica•2024•DOI: 10.3724/abbs.2023268
MicroRNAs (miRNAs) are small noncoding RNAs (ncRNAs) that play their roles in the regulation of physiological and pathological processes. Originally, it was assumed that miRNAs only modulate gene expression post-transcriptionally in the cytoplasm by inducing target mRNA degradation. However, with further research, evidence shows that mature miRNAs also exist in the cell nucleus, where they can impact gene transcription and ncRNA maturation in several ways. This review provides an overview of novel models of nuclear miRNA functions. Some of the models remain to be verified by experimental evidence, and more details of the miRNA regulation network remain to be discovered in the future.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2025137
DNA damage repair is a critical physiological process. The combined treatment of L-arginine (L-Arg) and 5-fluorouracil (5-FU) significantly inhibits cell proliferation, enhances nitric oxide (NO) production via inducible nitric oxide synthase (iNOS), and promotes the accumulation of reactive oxygen species (ROS). This heightened oxidative stress triggers DNA damage and apoptosis, as evidenced by a substantial increase in the Bax/Bcl-2 ratio; the activation of caspase-9, caspase-3, and PARP cleavage; and increased level of phosphorylated p53. Moreover, the combination treatment induces G2/M phase arrest, with a significant increase in p-H2AX (Ser 139) (known as γ-H2AX) expression, indicating extensive DNA damage. Mechanistically, the combined treatment modulates DNA damage response pathways by downregulating DNA-PKcs. Concurrently, it enhances the phosphorylation of ATM, ATR, CHK1, CHK2, and BRCA1. Additionally, the L-Arg and 5-FU combination downregulates PI3K/AKT signaling. AZD-7648 (a DNA-PKcs inhibitor) and LY294002 (a PI3K inhibitor) enhance p-ATM and p-ATR activation, resulting in elevated apoptosis and increased γ-H2AX expression. In contrast, the inhibition of ATM/ATR by CGK733 suppresses this response, reducing apoptosis and DNA damage signaling. Additionally, the ROS scavengers NAC and iNOS, when applied separately, restore p-AKT and DNA-PKcs expression; suppress the upregulation of p-ATM, p-ATR, and γ-H2AX; and ultimately reduce apoptosis. These findings are validated in a DEN-induced rat liver cancer model. In summary, 5-FU and L-Arg synergistically increase iNOS/NO-driven ROS accumulation, inducing γ-H2AX-marked DNA damage through dual modulation of repair pathways (inhibiting PI3K/AKT/DNA-PKcs while activating ATM/ATR), ultimately triggering p53-mediated G2/M arrest and apoptosis in hepatocellular carcinoma cells.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2025064
Hirschsprung’s disease (HSCR) is a congenital disorder characterized by the absence of enteric ganglion cells in the distal colon, resulting in functional intestinal obstruction. While genetic mutations and microenvironmental imbalances have been implicated in HSCR, the underlying molecular mechanisms are not fully understood. This study uses integrated quantitative proteomics and phosphoproteomics analyses to characterize the differential protein profiles and phosphorylation modifications associated with HSCR. These findings reveal significant dysregulation of the extracellular matrix (ECM) remodelling pathway, suggesting its potential involvement in HSCR pathogenesis. Notably, the deubiquitinating enzyme USP46 is found to be significantly reduced in the aganglionic segments of HSCR patients. Through IP-MS, GST pull-down, and co-immunoprecipitation assays, it is demonstrated that USP46 interacts with the transcription factor POU4F1. Mechanistically, USP46 stabilizes POU4F1 via deubiquitination, increasing its binding to the heparanase (HPSE) promoter and increasing HPSE expression, which in turn promotes ECM remodelling and neural cell migration. The role of the USP46-POU4F1-HPSE signaling axis in HSCR pathogenesis is confirmed via chromatin immunoprecipitation-qPCR, luciferase reporter assays, and transwell migration assays. This study elucidates a novel regulatory mechanism linking USP46-mediated protein stabilization to ECM dynamics and neural cell migration, offering new insights into HSCR pathogenesis and potential therapeutic targets.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2024195
PIWI-interacting RNAs (piRNAs), which associate with PIWI clade Argonaute proteins to form piRNA-induced silencing complexes (piRISCs) in germline cells, are responsible for maintaining genomic integrity and reproductive function through transcriptional or post-transcriptional suppression of transposable elements and regulation of protein-coding genes. Recent discoveries of crucial PIWI-piRNA functions in oogenesis and embryogenesis in golden hamsters suggest an indispensable role in female fertility that has been obscured in the predominant mouse model of PIWI-piRNA pathway regulation. In particular, studies of piRNA expression dynamics, functional redundancies, and compositional variations across mammal species have advanced our understanding of piRNA functions in male and, especially, female reproduction. These findings further support the use of hamsters as a more representative model of piRNA biology in mammals. In addition to discussing these new perspectives, the current review also covers emerging directions for piRNA research, its implications for female fertility, and our fundamental understanding of reproductive mechanisms.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2024138
Chemoresistance is the primary reason for poor prognosis in patients with pancreatic cancer (PC). Recent studies have indicated that ferroptosis may improve chemoresistance, but the underlying mechanisms remain unclear. In this study, significant upregulation of heat shock protein 90α (Hsp90α) expression is detected in the peripheral blood and tissue samples of patients with chemoresistant PC. Further studies reveal that Hsp90α promotes the proliferation, migration, and invasion of a chemoresistant pancreatic cell line (Panc-1-gem) by suppressing ferroptosis. Hsp90α competitively binds to Kelch-like ECH-associated protein 1 (Keap1), liberating nuclear factor erythroid 2-related factor 2 (Nrf2) from Keap1 sequestration. Nrf2 subsequently translocates into the nucleus and activates the glutathione peroxidase 4 (GPX4) pathway, thereby suppressing ferroptosis. This process further worsens the chemoresistance of PC cells. This study provides valuable insight into potential molecular targets to overcome chemoresistance in PC. It sheds light on the intricate mechanisms linking Hsp90α and ferroptosis to chemoresistance in PC and provides a theoretical foundation for the development of novel therapeutic strategies.
Acta Biochimica et Biophysica Sinica•2024•DOI: 10.3724/abbs.2024015
Traditional Chinese medicine (TCM) has been used to treat triple-negative breast cancer (TNBC), a breast cancer subtype with poor prognosis. Clinical studies have verified that the Sanyingfang formula (SYF), a TCM prescription, has obvious effects on inhibiting breast cancer recurrence and metastasis, prolonging patient survival, and reducing clinical symptoms. However, its active ingredients and molecular mechanisms are still unclear. In this study, the active ingredients of each herbal medicine composing SYF and their target proteins are obtained from the Traditional Chinese Medicine Systems Pharmacology database. Breast cancer-related genes are obtained from the GeneCards database. Major targets and pathways related to SYF treatment in breast cancer are identified by analyzing the above data. By conducting molecular docking analysis, we find that the active ingredients quercetin and luteolin bind well to the key targets KDR1, PPARG, SOD1, and VCAM1. In vitro experiments verify that SYF can reduce the proliferation, migration, and invasion ability of TNBC cells. Using a TNBC xenograft mouse model, we show that SYF could delay tumor growth and effectively inhibit the occurrence of breast cancer lung metastasis in vivo. PPARG, SOD1, KDR1, and VCAM1 are all regulated by SYF and may play important roles in SYF-mediated inhibition of TNBC recurrence and metastasis.
Acta Biochimica et Biophysica Sinica•2024•DOI: 10.3724/abbs.2024188
DNA replication stress is a significant contributor to spontaneous DNA damage and genome instability. While the impact of p53 deficiency on increasing DNA replication stress is known, the specific molecular mechanism underlying this phenomenon remains poorly understood. This study explores how p53 deficiency induces DNA replication stress by activating mTORC1 through R-loop formation, which is facilitated by the upregulation of RNR. Research has shown that p53 deficiency results in increased γH2AX expression and a higher mutation rate in the HPRT gene. Interestingly, these effects can be alleviated by rapamycin, an mTORC1 inhibitor. Additionally, rapamycin reduces the abundance of R-loop structures in p53KO cells, which is linked to mTORC1’s regulation of ribonucleotide reductase (RNR) level. These findings suggest that p53 deficiency-induced DNA replication stress relies on mTORC1 activation, with the upregulation of RNR expression and R-loop formation. Overall, this study underscores the importance of R-loops in mTORC1 activation-dependent DNA replication stress triggered by p53 deficiency.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2024170
Doxorubicin (Dox) is widely utilized in the clinical treatment of various cancers. Despite its efficacy, Dox induces numerous adverse effects in humans with significant cardiotoxicity, posing a major limitation to its use. Saussurea involucrata injection (SII), derived from Saussurea involucrata, exhibits notable anti-inflammatory and anti-oxidative stress properties. However, its potential protective effects against Dox-induced cardiotoxicity (DIC) remain unexplored. In this study, we investigate the ability of SII to mitigate DIC and elucidate the underlying mechanisms through experimental research and network pharmacology analysis. Results from both in vitro and in vivo experiments reveal that SII treatment significantly improves Dox-induced cardiac dysfunction, reducing pathological alterations and fibrosis in cardiomyocytes. Moreover, SII has cardioprotective effects by diminishing the inflammation, oxidative stress, and apoptosis triggered by Dox. Network pharmacological analysis further shows that SII downregulates P53 protein expression by activating the AKT/MDM2 signaling pathway, thus attenuating DIC. In conclusion, this study confirms that SII mitigates DIC through downregulation of the AKT/MDM2/P53 signaling pathway, suggesting a promising therapeutic strategy for alleviating DIC.
Acta Biochimica et Biophysica Sinica•2024•DOI: 10.3724/abbs.2023279
RNA terminal phosphorylase B (RTCB) has been shown to play a significant role in multiple physiological processes. However, the specific role of RTCB in the mouse colon remains unclear. In this study, we employ a conditional knockout mouse model to investigate the effects of RTCB depletion on the colon and the potential molecular mechanisms. We assess the efficiency and phenotype of Rtcb knockout using PCR, western blot analysis, histological staining, and immunohistochemistry. Compared with the control mice, the Rtcb-knockout mice exhibit compromised colonic barrier integrity and prominent inflammatory cell infiltration. In the colonic tissues of Rtcb-knockout mice, the protein levels of TNF-α, IL-8, and p-p65 are increased, whereas the levels of IKKβ and IκBα are decreased. Moreover, the level of GSK3β is increased, whereas the levels of Wnt3a, β-catenin, and LGR5 are decreased. Collectively, our findings unveil a close association between RTCB and colonic tissue homeostasis and demonstrate that RTCB deficiency can lead to dysregulation of both the NF-κB and Wnt/β-catenin signaling pathways in colonic cells.
Acta Biochimica et Biophysica Sinica•2024•DOI: 10.3724/abbs.2023275
Acute liver failure (ALF) is a significant global issue with elevated morbidity and mortality rates. There is an urgent and pressing need for secure and effective treatments. Ferroptosis, a novel iron-dependent regulation of cell death, plays a significant role in multiple pathological processes associated with liver diseases, including ALF. Several studies have demonstrated that mesenchymal stem cells (MSCs) have promising therapeutic potential in the treatment of ALF. This study aims to investigate the positive effects of MSCs against ferroptosis in an ALF model and explore the underlying molecular mechanisms of their therapeutic function. Our results show that intravenously injected MSCs protect against ferroptosis in ALF mouse models. MSCs decrease iron deposition in the liver of ALF mice by downregulating hepcidin level and upregulating FPN1 level. MSCs labelled with Dil are mainly observed in the hepatic sinusoid and exhibit colocalization with the macrophage marker CD11b fluorescence. ELISA demonstrates a high level of IGF1 in the CCL4+MSC group. Suppressing the IGF1 effect by the PPP blocks the therapeutic effect of MSCs against ferroptosis in ALF mice. Furthermore, disruption of IGF1 function results in iron deposition in the liver tissue due to impaired inhibitory effects of MSCs on hepcidin level. Our findings suggest that MSCs alleviate ferroptosis induced by disorders of iron metabolism in ALF mice by elevating IGF1 level. Moreover, MSCs are identified as a promising cell source for ferroptosis treatment in ALF mice.
Stem Cell Research & Therapy•2026•DOI: 10.1186/s13287-026-04913-w
Extracorporeal cardiac shock wave (ECSW) therapy has been shown to enhance endothelial colony-forming cell (ECFC) function, but its potential as a preconditioning strategy for myocardial infarction (MI) therapy remains undefined. This study evaluated the efficacy and mechanism of intravenously delivered ECSW-preconditioned ECFCs (SW-ECFCs) in a rat MI model. ECFCs were isolated from ApoE-/- rat bone marrow and characterized. RNA sequencing revealed significant enrichment of the PI3K/AKT pathway in SW-ECFCs. In vitro assays demonstrated that ECSW enhanced ECFC migration, tube formation, proliferation, and VEGF-A secretion while reducing apoptosis; these effects were interrogated using the PI3K/AKT inhibitor LY294002. An acute MI model was established via left anterior descending coronary artery ligation in ApoE-/- rats, randomized into four groups: MI + PBS, MI + ECFCs, MI + SW-ECFCs, and MI + LY294002-pretreated SW-ECFCs (LY-SW-ECFCs), with sham controls. Comprehensive evaluations included echocardiography, serum injury biomarkers, TTC staining, histopathology (H&E, Masson), immunohistochemistry for cardiomyocyte apoptosis and p-eNOS, immunofluorescence for ECFC homing and vascular markers (CD31, α-SMA, VEGF-A), nitric oxide measurement, and Western blot analysis of PI3K/AKT signaling proteins. Results confirmed that ECSW preconditioning significantly improved cardiac function and structural repair, mediated primarily through PI3K/AKT pathway activation, augmenting cell homing, paracrine activity, and survival. This work presents a novel and promising strategy for cardiac regeneration.
Stem Cell Research & Therapy•2026•DOI: 10.1186/s13287-026-05061-x
Osteoporosis is characterized by impaired bone formation relative to resorption, yet the molecular drivers of osteoblast dysfunction remain incompletely defined. Cadherin 19 (CDH19), located at chromosome 18q22-q23, has been linked to 18q deletion syndromes presenting with skeletal deformities, but its role in bone homeostasis was previously unknown. Using a Cre-loxP conditional knockout model, we demonstrate that CDH19 deletion in mice significantly reduces bone mass, with decreases in bone density, trabecular number, and bone volume fraction. Osteoblasts isolated from CDH19 knockout mice exhibit suppressed proliferation and osteogenic differentiation, as evidenced by EdU labeling, qPCR, alkaline phosphatase and alizarin red S staining, and Western blot. RNA sequencing and subsequent immunofluorescence and Western blot analyses reveal that the PI3K/AKT signaling pathway is markedly inhibited in CDH19-deficient osteoblasts. Administration of the PI3K/AKT agonist 740Y-P partially rescues the osteogenic differentiation deficit both in vitro and in vivo. These findings establish CDH19 as a critical regulator of osteoblast function through PI3K/AKT signaling and identify it as a potential therapeutic target for bone diseases such as osteoporosis.
Stem Cell Research & Therapy•2026•DOI: 10.1186/s13287-026-04902-z
Inherited cardiomyopathy (ICM) arises from genetic mutations that compromise myocardial structure and function, with FHOD3 mutations linked to hypertrophic, dilated, and left ventricular noncompaction cardiomyopathies. The molecular mechanisms driving FHOD3 deficiency-induced cardiomyopathy remain undefined. A FHOD3 knockout (FHOD3-/-) human embryonic stem cell line was generated via CRISPR/Cas9 and differentiated into cardiomyocytes (hESC-CMs). Sarcomere architecture, calcium handling, mitochondrial function, and contractility were assessed using immunofluorescence, electron microscopy, Seahorse metabolic analysis, and high-definition video analysis. Transcriptomic sequencing identified differentially expressed genes and enriched pathways. FHOD3-deficient hESC-CMs displayed pronounced sarcomere disorganization and degradation, impaired calcium handling, and compromised mitochondrial function, culminating in reduced contractility. Transcriptomic profiling revealed significant downregulation of sarcomere-related and calcium-handling genes, with enrichment in cardiomyopathy and calcium signaling pathways. FHOD3 deficiency triggered phosphorylation of CaMKII at Thr286, a key regulator of cardiac hypertrophy and remodeling, contributing to heart failure progression. Treatment with the myosin activator Omecamtiv mecarbil (OM) partially restored contractility without affecting calcium handling, highlighting its therapeutic potential. This human-derived loss-of-function model establishes CaMKII activation as a critical factor in FHOD3-deficiency-induced heart failure and provides a robust platform for discovering novel therapeutic agents.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2026054
The anti-inflammatory properties of the traditional herb Salvia miltiorrhiza Bunge are well-established, yet its precise mechanism of action in ulcerative colitis (UC) remains unclear. Herein, we evaluate the therapeutic potential of four major tanshinones–tanshinone IIA (Tan IIA), miltirone, neocryptotanshinone, and dihydrotanshinone I–in a murine dextran sulfate sodium (DSS)-induced colitis model. Our results show that tanshinones effectively alleviate disease severity, suppress systemic and local inflammation, and restore intestinal barrier integrity. Integrated multi-omics analysis reveals that the therapeutic efficacy originates from a comprehensive reprogramming of the gut microbiota-metabolite axis. Specifically, tanshinones reverse colitis-associated dysbiosis and rectify metabolic disturbances in linoleic acid metabolism, bile acid biosynthesis, and amino acid utilization. Correlation network analysis identifies key functional modules linking beneficial microbes (e.g., Akkermansia) to anti-inflammatory lipid mediators and associating pathobionts (e.g., Desulfovibrio) with disrupted bile acid metabolism. Notably, supplementation with Akkermansia muciniphila synergizes with Tan IIA to amplify barrier restoration and metabolic normalization. Our findings establish that tanshinones ameliorate UC through microbiota-driven metabolic reprogramming, wherein the restructured microbial community actively shapes a therapeutic metabolic output. This work elucidates a metabolite-mediated mechanism of action and positions tanshinones as promising microbiome-targeting therapeutics for inflammatory bowel disease.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2025196
Human embryonic kidney (HEK) 293 cells are widely used for recombinant protein production because of their efficient posttranslational modification capabilities. However, their large-scale culture is often limited by metabolic stress and early apoptosis, leading to insufficient protein yields. In this study, we aim to increase protein expression through the coordinated modulation of metabolic and apoptotic pathways. Using CRISPR/Cas9 technology, we target and knockout the genes of ornithine decarboxylase antizyme 1 (OAZ1), which regulates polyamine metabolism, and caspase 8-associated protein 2 (CASP8AP2), an apoptosis-related protein. We successfully construct an OAZ1/CASP8AP2 double-knockout HEK293 cell line. Following transfection with the knockout vector and screening of single-cell clones, multiple levels of validation confirm the successful gene knockout. The results show that the double-knockout cells exhibit significantly reduced apoptosis rates. Furthermore, the production of recombinant secreted alkaline phosphatase (SEAP) and vitronectin (VN) increases by 2.1 folds and 2.9 folds, respectively, compared with those in wild-type cells. Metabolic profiling reveals that the cell cycle is arrested in the G1/G0 phase, accompanied by increased specific consumption and production rates of key metabolites. This study demonstrates that concurrent inhibition of apoptosis and optimization of metabolism effectively enhances recombinant protein production in HEK293 cells, suggesting a novel strategy for improving HEK293 cell-based expression.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21265
BACKGROUND: In recent years, the interdisciplinary application potential of patch-clamp technique in traditional Chinese medicine research has gradually emerged, but a systematic summary of its applications in this field has not yet been conducted. OBJECTIVE: To visualize the application of patch-clamp technology in traditional Chinese medicine field through CiteSpace knowledge map analysis, and to reveal the progress and trends of this technology in the modernization research of traditional Chinese medicine. METHODS: The literature sources included CNKI, VIP, WanFang, PubMed and Web of Science Core Collection database. The computer-assisted literature search was conducted to build a database of patch-clamp technology applications in traditional Chinese medicine field from database inception to September 2024. The authors, institutions, and keywords were subjected to visual analysis and knowledge map drawing using CiteSpace 6.3.R1 software and bibliometric methods. RESULTS AND CONCLUSION: (1) A total of 819 articles were included, with 968 authors. The First Affiliated Hospital of Henan University of Chinese Medicine was the institution with the most publications in the Chinese database, while Harbin Medical University was the institution with the most publications in the English database. (2) The research directions, keyword clustering, and emergence analysis of each institution showed that the application of patch-clamp technique in traditional Chinese medicine mainly focused on cardiovascular electrophysiology, pharmacology of Chinese materia medica, and nervous system electrophysiology. (3) The development of patch-clamp technique in traditional Chinese medicine generally presented an evolutionary path of 'basic mechanism → target deepening → clinical translation', reflecting a transformation from a single technical tool to a multidisciplinary intersection platform. (4) It is suggested that the core influence of authors needs further improvement, and cross-regional cooperation among research institutions is insufficient, so cross-regional cooperation should be strengthened. Current research techniques are single and research content is thin; future research should integrate multiple technologies, cross disciplines, enrich research content, and expand research directions, providing evidence support for in-depth exploration of the internal mechanisms of traditional Chinese medicine.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21276
BACKGROUND: Glucagon-like peptide 1 receptor agonists, as novel drug candidates for the treatment of neurodegenerative diseases, have achieved breakthrough progress in clinical research on Alzheimer’s disease, with drugs such as Semaglutide advancing to phase III clinical trials. However, there remains a significant knowledge gap regarding the molecular mechanism of neuroprotective effects of these drugs. OBJECTIVE: To innovatively integrate multi-omics analysis techniques and network pharmacology methods, to systematically analyze the intersection network between the gene lineage related to Alzheimer’s disease pathology and the potential targets of ticagrelor, to identify key regulatory genes, and to verify their molecular mechanisms through in vitro and in vivo experiments. METHODS: A multi-dimensional research strategy was adopted: (1) Constructing the differential expression gene profile of Alzheimer’s disease using the DisGeNET database that covers various disease-related genomics. (2) Obtaining the structure of Tirzepatide from PubChem database with bioactive molecules and screening potential targets. (3) Conducting Gene Ontology (GO) functional annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis using DAVID database. (4) Constructing protein-protein interaction network using STRING database and Cytoscape 3.9.1, and screening key genes via topological network analysis. (5) Cell-level verification: HT22 cells were divided into control group, model group (treated with β-amyloid 1-42 oligomers for 36 h to establish an in vitro AD model), and treatment group (pretreated with β-amyloid 1-42 oligomers for 24 h, then co-treated with ticagrelor for 12 h). Western blot was used to analyze the protein expression of angiotensin II type 2 receptor (AGTR2), and ELISA was used to detect the expression levels of synaptic markers such as synaptophysin 1 and postsynaptic density protein 95. (6) Animal experiments: Three groups were used: control group (WT C57BL/6 mice, intraperitoneal injection of saline), model group (3xTg mice, intraperitoneal injection of saline), and treatment group (3xTg mice, intraperitoneal injection of 20 nmol/L ticagrelor), all administered every other day for a total of 15 doses. Morris water maze was used to analyze cognitive behavioral improvements in AD model mice; Western blot was used to quantitatively analyze the expression of β-amyloid (6E10) and phosphorylated Tau protein (P-tau-181). RESULTS AND CONCLUSION: (1) A total of 3,397 AD-related genes were screened from DisGeNET database; 10 key genes with the highest connectivity were identified based on protein association: AGTR2, NTSR1, NTSR2, GHSR, C5AR1, C3AR1, OPRM1, SSTR2, OPRD1, STAT3. GO enrichment and KEGG pathway analysis suggested that ticagrelor may improve AD by enhancing neuroreceptor-ligand function. (2) Cell experiments suggested that ticagrelor may exert therapeutic effects by improving synaptic function in AD, and AGTR2 may be a potential target of ticagrelor in treating AD. (3) Animal experiments indicated that ticagrelor improved cognitive ability in 3xTg mice, and ameliorated abnormal β-amyloid deposition and Tau protein phosphorylation in the brain of 3xTg mice. (4) Conclusion: The study reveals that AGTR2 is a key molecular target of ticagrelor in the pathological process of AD, and ticagrelor may treat AD by regulating AGTR2-mediated synaptic function improvement.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21293
BACKGROUND: As the primary organ for arsenic metabolism in the body, the liver has become a focal point for research on the mechanisms of arsenic toxicity. OBJECTIVE: To investigate the role of pregnane X receptor in sodium arsenite-induced oxidative stress and inflammatory injury in human normal hepatocytes. METHODS: Human normal hepatocyte MIHA cells were exposed to 0 (control), 10, 20, 30 μmol/L sodium arsenite for 48 hours. Changes in cell morphology were observed. Cell viability was measured via the cell counting kit-8 assay. Intracellular reactive oxygen species levels were detected using fluorescence probe staining combined with a microplate reader. Malondialdehyde levels were measured by thiobarbituric acid method. Glutathione reductase activity was detected by NADPH method. Total superoxide dismutase activity was measured by WST-8 method. Levels of interleukin-6, interleukin-1β, and tumor necrosis factor-α in cell supernatant were detected by ELISA. mRNA expression of pregnane X receptor and cytochrome P450 3A4 enzyme was detected by qRT-PCR. Protein expression of pregnane X receptor, cytochrome P450 3A4, nuclear factor-κB p65, nuclear factor-κB p-p65, proliferating cell nuclear antigen, interleukin-6, interleukin-1β, tumor necrosis factor-α, nuclear factor-κB inhibitor protein α, cyclooxygenase-2, p-nuclear factor-κB inhibitor protein α, nuclear factor erythroid 2-related factor 2, Keap1, and p-nuclear factor erythroid 2-related factor 2 was detected by western blot. RESULTS AND CONCLUSION: Compared with the control group, cells in all sodium arsenite groups showed unclear cell membrane boundaries, reduced cytoplasm, decreased cell fusion rate, and widened intercellular spaces. Compared with the control group, intracellular reactive oxygen species and malondialdehyde levels were increased (P < 0.05), levels of interleukin-6, interleukin-1β, and tumor necrosis factor-α in cell supernatant were increased (P < 0.05), protein expression of p-nuclear factor-κB inhibitor protein α, nuclear factor-κB p-p65, nuclear factor-κB p65, tumor necrosis factor-α, and interleukin-1β were increased (P < 0.05), cell viability was decreased (P < 0.05), protein expression of proliferating cell nuclear antigen, nuclear factor erythroid 2-related factor 2, p-nuclear factor erythroid 2-related factor 2 and total superoxide dismutase activity were decreased (P < 0.05), and mRNA and protein expression of pregnane X receptor and cytochrome P450 3A4 enzyme were decreased (P < 0.05). Compared with the control group, glutathione reductase activity was decreased in 20 and 30 μmol/L sodium arsenite groups (P < 0.05), and protein expression of Keap1, interleukin-6, and cyclooxygenase-2 was increased (P < 0.05). These results indicate that sodium arsenite may induce oxidative stress and inflammatory injury in hepatocytes by downregulating pregnane X receptor expression, inhibiting the nuclear factor erythroid 2-related factor 2 antioxidant pathway, and activating the nuclear factor-κB inflammatory pathway, while also inhibiting the expression of the drug-metabolizing enzyme cytochrome P450 3A4.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21412
BACKGROUND: Currently, most studies on rigid post-traumatic thoracolumbar kyphosis focus on overall sagittal balance and surgical intervention, while the effect of the disease duration on the change of lumbar compensatory curvature and degeneration is still unclear. OBJECTIVE: To explore the effect of the disease duration on lumbar degeneration and the potential mechanism of rigid post-traumatic thoracolumbar kyphosis in patients with rigid post-traumatic thoracolumbar kyphosis, and provide a basis for optimizing treatment strategies. METHODS: Clinical and imaging data from 79 rigid post-traumatic thoracolumbar kyphosis patients were retrospectively analyzed. The patients were divided into two groups according to the disease duration: Patients with a disease duration of ≤ 5 years were categorized as group A (n=40), and those with > 5 years as group B (n=39). X-ray images were used to measure the local kyphosis angle of the injured vertebra, the height of the posterior walls of the injured vertebra and adjacent vertebrae, lumbar lordosis, the intervertebral space angle for each lumbar segment, and sacral slope. The Weishaupt-CT classification system was employed to assess lumbar facet joint degeneration. Pfirrmann-MRI grading was used to evaluate intervertebral disc degeneration. Clinical outcomes including visual analog scale for back pain, Oswestry Disability Index, SRS-22 score, and American Spinal Injury Association impairment scale were compared between groups. The influence of disease duration on clinical symptoms and imaging features was analyzed. RESULTS AND CONCLUSION: (1) There were no significant differences in age, sex, visual analog scale score, fracture site, fracture morphology, or American Spinal Injury Association grade between the two groups (P > 0.05). The SRS-22 subscore was significantly higher in group A than in group B (P < 0.05), while the Oswestry Disability Index was significantly higher in group B than in group A (P < 0.05). (2) The local kyphosis angle, lumbar lordosis, and L4/5 intervertebral space angle were significantly greater in group B than in group A (P < 0.05). (3) There were no significant differences in L1/2, L2/3, L3/4, L5/S1 intervertebral space angles and sacral slope between groups (P > 0.05). (4) The degree of facet joint degeneration at L3/4, L4/5, and L5/S1 was significantly more severe in group B than in group A (P < 0.05). The degree of intervertebral disc degeneration at L2/3, L3/4, L4/5, and L5/S1 was significantly more severe in group B than in group A (P < 0.05). (5) Pearson correlation analysis showed that within group B, disease duration was positively correlated with local kyphosis angle and lumbar lordosis (r=0.335, 0.418, P < 0.05). (6) In patients with rigid post-traumatic thoracolumbar kyphosis, long-term compensation leads to increased lumbar lordosis and accelerated lumbar degeneration. The L4/5 segment is the main compensatory segment in lumbar curvature compensation, and special attention should be paid to the correction of lower lumbar curvature during surgical correction.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21413
BACKGROUND: Recently, percutaneous minimally invasive pedicle screw fixation systems have become a popular treatment option for thoracolumbar fractures due to ongoing innovation in orthopedic internal fixation devices. Currently, while many studies compare the effectiveness of open pedicle screw fixation through or across the fractured vertebra, fewer studies compare these two methods when using navigation-assisted percutaneous minimally invasive techniques. OBJECTIVE: To compare the clinical efficacy of percutaneous minimally invasive pedicle screw fixation in the treatment of thoracolumbar fracture through and across the injured vertebra. METHODS: A retrospective analysis was performed on 67 patients with single level thoracolumbar fracture without spinal cord nerve injury who were treated in the Department of Spine Surgery, Affiliated Hospital of Yan'an University from October 2021 to June 2023. All of them were treated with percutaneous pedicle screw fixation with the assistance of computer navigation, and were followed up to 6 months after surgery. The injured vertebrae were divided into transinjured vertebrae group (n=35) and cross-injured vertebrae group (n=32). RESULTS AND CONCLUSION: ① There were no significant differences in preoperative general data between the two groups (P > 0.05), indicating comparability. ② Intragroup comparison: In both groups, the anterior vertebral height ratio, Cobb angle, visual analog scale score, and Oswestry disability index at 7 days and 6 months after surgery were significantly improved compared with preoperative values (P < 0.05). ③ Intergroup comparison: The operation time, intraoperative blood loss, incision length, and intraoperative fluoroscopy times in the cross-injured vertebrae group were significantly less than those in the trans-injured vertebrae group (P < 0.05), while there was no significant difference in hospital stay between the two groups (P > 0.05). At 7 days and 6 months after surgery, there were no significant differences in anterior vertebral height ratio, Cobb angle, visual analog scale score, and Oswestry disability index between the two groups (P > 0.05). ④ It is suggested that for patients with single-level thoracolumbar fractures without spinal cord nerve injury, compared with trans-injured vertebra fixation, percutaneous minimally invasive pedicle screw fixation across the injured vertebra has advantages of shorter operation time, less bleeding, smaller surgical incision, and fewer fluoroscopy times. In the medium-term follow-up period, both trans-injured and cross-injured percutaneous minimally invasive fixation can effectively restore and maintain the height of the injured vertebra, correct kyphotic deformity, rebuild spinal stability, and relieve pain, achieving excellent clinical results. For patients with AO type A thoracolumbar fractures with normal body mass index, without spinal cord nerve injury and posterior ligamentous complex injury, the less traumatic cross-injured vertebra percutaneous pedicle screw fixation is recommended.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21418
BACKGROUND: Clinically, due to the special anatomical characteristics and internal trabecular bone distribution of the scaphoid, the treatment effect of fractures is generally poor, often leading to nonunion and ischemic necrosis, which in turn causes wrist arthritis and loss of function. OBJECTIVE: To scan scaphoid specimens using Micro CT technology, analyze their internal microstructure characteristics, measure the trabecular bone microstructure parameters in each region, and discover regional differences in scaphoid trabecular bone, aiming to provide a scientific basis for the prevention, treatment, and fracture mechanism research of scaphoid fractures. METHODS: Bilateral scaphoid bones (10 cases) from 5 adult cadaver specimens were scanned by Micro CT. By selecting and reconstructing trabecular bone in three regions of interest (tubercle, waist, and body), the internal micromorphological characteristics of the scaphoid were observed in detail, and the differences in trabecular bone microstructure parameters among regions were measured and compared. RESULTS AND CONCLUSION: (1) Micro CT images showed that the cortical bone on the surface of the scaphoid was relatively thin, and the interior was filled with complex trabecular bone microstructure; the lamellar trabecular bone near the cortical bone was relatively dense, extending inward into rod-like trabecular bone. From sagittal, coronal, and transverse sections, the trabecular bone distribution in the waist was relatively sparse, while that in the body and tubercle was denser. (2) There were significant differences in bone volume fraction, bone surface area, bone surface area to tissue volume ratio, trabecular separation, trabecular number, trabecular connectivity, trabecular connection density, fractal dimension, bone mineral density, and bone mineral content of the scaphoid tubercle between left and right sides (P < 0.05). There were no significant differences in the trabecular bone microstructure parameters of the waist and body between left and right sides (P > 0.05). (3) There were significant differences in bone volume, bone volume fraction, bone surface area, bone surface area to tissue volume ratio, bone surface area to bone volume ratio, bone mineral density, and bone mineral content between the body and the tubercle/waist (P < 0.05). There was a significant difference in trabecular thickness between the body and the tubercle (P < 0.05). There were significant differences in trabecular separation and fractal dimension among the body, tubercle, and waist (P < 0.05). There were significant differences in trabecular number, trabecular connectivity, and trabecular connection density between the waist and the tubercle/body (P < 0.05). There were no significant differences in tissue volume and degree of anisotropy among the body, tubercle, and waist (P > 0.05). (4) The results showed that the trabecular bone microstructure parameters of the scaphoid had regional differences, among which the waist had lower bone density and strength, making it the most prone to fracture. This finding provides a theoretical basis for understanding the fracture mechanism of the scaphoid from the perspective of trabecular bone microstructure. At the same time, the trabecular bone structure characteristics of different parts of the scaphoid revealed in this study also provide a theoretical basis for designing targeted internal fixation instruments.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21395
BACKGROUND: With the widespread application of total knee arthroplasty in China, non-infectious prosthesis loosening has become one of the main reasons for postoperative revision. For complex loosening cases, traditional revision techniques are relatively complex and difficult. The application of artificial intelligence-assisted preoperative planning combined with Sleeve extension rods and mobile bearing tray prostheses provides a new solution for precisely reconstructing joint stability and mechanical alignment, which is expected to improve the long-term outcomes of revision surgeries. OBJECTIVE: To explore the mid-and early-term clinical efficacy of revision surgery for non-infectious total knee prosthesis loosening using Sleeve extension rods combined with mobile bearing tray prostheses under the assistance of artificial intelligence-assisted preoperative planning. METHODS: A retrospective analysis was conducted on 17 patients with non-infectious prosthesis loosening after total knee arthroplasty in Department of Orthopedics and Traumatology, Jiangsu Provincial Hospital of Traditional Chinese Medicine from January 2021 to September 2024. There were 6 males and 11 females, aged 59-81 years (mean 72.06±6.10 years). The affected side was left in 8 cases and right in 9 cases. The duration of prosthesis use ranged from 2 to 22 years (mean 10.53±4.60 years). All cases were revisions after primary arthroplasty. Revision reasons included periprosthetic osteolysis with liner wear in 15 cases, femoral condyle old fracture causing loosening in 1 case, and tibial plateau prosthesis fracture in 1 case. According to AORI classification, there were 13 cases of type IIB and 4 cases of type IIA. The artificial intelligence-designed prosthesis sizes were recorded and compared with the actual intraoperative sizes. Visual analog scale (VAS) score, American Knee Society knee score, hip-knee-ankle angle, and knee range of motion were compared preoperatively, at 1 week, 6 months, and 12 months postoperatively to evaluate surgical efficacy. RESULTS AND CONCLUSION: (1) Except for one patient with poor incision healing at 1 month postoperatively, all other patients recovered well without adverse events such as deep vein thrombosis, infection, periprosthetic fracture, or prosthesis loosening. (2) The follow-up period ranged from 6 to 41 months (mean 23.63±12.50 months). At the last follow-up, 2 patients had slight soreness and discomfort after activity, and 1 patient had obvious pain during activity. (3) At the last follow-up after revision, resting pain, exercise pain VAS scores, affected side range of motion, hip-knee-ankle angle, and American Knee Society knee score were significantly improved compared with preoperative values (P < 0.01). (4) The matching rate of artificial intelligence preoperative design for femoral condyle and tibial plateau prostheses was 85%, and the matching rate for other components was 62%. (5) The use of Sleeve+ extension rod combined with MBT prosthesis for non-infectious knee revision can effectively correct joint alignment, fill bone defects, improve pain and knee range of motion, and enhance patients' quality of life, with good early and mid-term efficacy. Artificial intelligence preoperative planning generally helps improve surgical accuracy, reduce revision difficulty, minimize risks, and promote postoperative recovery.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21312
BACKGROUND: Although physcion has been shown to have protective effects against osteoporosis, the exact mechanism is not fully understood. OBJECTIVE: Through multidimensional analysis of the regulatory effect of physcion on the AKT signaling pathway, the molecular mechanism of its regulation on osteoclast induced differentiation and osteogenic function induced differentiation is revealed. METHODS: (1) RAW264.7 cells and C3H10T1/2 cells were cultured in vitro and subsequently exposed to 0, 10, 20, 30, 40, 50, and 60 µmol/L physcion, respectively. The cytotoxicity of physcion was detected by cell counting kit-8 assay. (2) RAW264.7 cells and C3H10T1/2 cells were treated with different concentrations (0, 20, 40 µmol/L) of physcion during osteoclast and osteoblast differentiation, respectively. Differentiation ability was assessed by qPCR, Western Blot, and alkaline phosphatase staining. (3) Network pharmacology was used to analyze the regulation of physcion on osteoclast differentiation and related signaling pathways, and molecular docking was performed for target proteins. (4) Western Blot was used to verify the phosphorylation level of AKT in the downstream target signaling pathway AKT axis regulated by physcion. RESULTS AND CONCLUSION: (1) At concentrations of 0-60 µmol/L, cell viability in all groups was greater than 90%, indicating no significant cytotoxicity. (2) Physcion significantly inhibited the expression of osteoclast differentiation-related genes, with Acp5, CTSK, DC-STAMP, and Nfatc1 showing downregulation, but had no significant effect on osteoblast differentiation-related genes COL1A1, Runx2, OSX expression or alkaline phosphatase staining intensity. (3) Network pharmacology and molecular docking suggested that physcion affects osteoclast differentiation and regulates the PI3K-AKT pathway, with a binding energy of -10.72 kJ/mol to AKT1, indicating strong binding activity. (4) During osteoclast differentiation, the p-AKT/AKT ratio in RAW264.7 cells increased (n=3, P=0.0063), while physcion decreased this ratio. These findings indicate that physcion inhibits osteoclast differentiation by regulating the AKT signaling pathway, thereby modulating bone homeostasis.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21362
BACKGROUND: With an increasing understanding of the health benefits of exercise, research on the mechanisms of exercise intervention has become a focal point. Traditional studies rely on in vivo animal models or multi-omics techniques to indirectly infer exercise intervention mechanisms, but the research is not in-depth enough, and many disease models cannot achieve the prescribed exercise intensity. Therefore, in vitro cell-based exercise environment simulation techniques are of particular significance. Existing technologies primarily focus on the replication of single signals, failing to comprehensively simulate the interaction of multi-dimensional signals during exercise, which limits the understanding of exercise adaptation mechanisms. OBJECTIVE: To explore the technological advancements in in vitro cell-based exercise environment simulation, analyze the advantages of existing signal simulation techniques, and propose a new framework integrating multi-dimensional signals to promote the precise replication of exercise mechanisms and application research in related fields. METHODS: This study conducted a search in the PubMed and Web of Science databases using keywords such as Exercise, Physiology, Molecular Signals, Myokines, Exerkines, etc. After initial screening and removal of duplicates, 5,046 relevant articles were identified, and 99 were finally included after further screening. RESULTS AND CONCLUSION: Existing in vitro cell exercise simulation techniques have made some progress in simulating specific attributes of exercise (e.g., mechanical stretching, electrical signals), but they still fail to fully replicate the multi-dimensional signal interactions during exercise. By integrating multiple signals such as mechanical forces, electrophysiological stimuli, and biological factors, future simulation technologies are expected to more realistically reproduce the effects of exercise on cellular metabolism, gene expression, and phenotypic remodeling, providing a more precise experimental platform for studying exercise mechanisms. Furthermore, innovations and optimizations in in vitro exercise simulation technologies will provide important support for sports medicine, drug development, and regenerative medicine.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21347
BACKGROUND: Rutin can effectively prevent osteoporosis, but its mechanism of action remains unclear. OBJECTIVE: To investigate the effect of rutin on osteogenesis of MC3T3-E1 cells under the action of neutrophil extracellular traps. METHODS: (1) Human myeloid leukemia dHL60 cells were stimulated with phorbol 12-myristate 13-acetate to induce neutrophil extracellular trap formation. dHL60 cells were divided into 4 groups: control group received Hank's balanced salt solution; the other three groups received 50 nmol/L phorbol 12-myristate 13-acetate; the latter two groups additionally received 250 μmol/L rutin or 250 μmol/L rutin plus 5 U/mL DNase I. Apoptosis of dHL60 cells was detected by flow cytometry; mRNA and protein expression of neutrophil extracellular trap marker genes were detected by RT-qPCR and western blot. (2) MC3T3-E1 cells were divided into 6 groups: control group received Hank's balanced salt solution; the other five groups received 50 nmol/L phorbol 12-myristate 13-acetate; dHL60 cells and 50 nmol/L phorbol 12-myristate 13-acetate; 100 μmol/L rutin; dHL60 cells, 50 nmol/L phorbol 12-myristate 13-acetate, and 250 μmol/L rutin; dHL60 cells, 50 nmol/L phorbol 12-myristate 13-acetate, 250 μmol/L rutin, and 5 U/mL DNase I. Apoptosis of MC3T3-E1 cells under neutrophil extracellular traps was detected by flow cytometry; alkaline phosphatase staining and alizarin red staining were used to determine osteogenic and mineralization abilities; RT-qPCR and western blot were used to detect osteogenic-related gene and protein expression. RESULTS AND CONCLUSION: (1) Compared with the blank control group, rutin significantly inhibited the mRNA and protein expression of protein arginine deiminase 4, myeloperoxidase, and neutrophil elastase in dHL60 cells (P < 0.000 1); compared with the rutin group, the combination of rutin and DNase I had a more significant downregulation effect (P < 0.05), indicating that rutin can significantly inhibit neutrophil extracellular trap formation. (2) After inducing neutrophil extracellular traps from dHL60 and co-culturing with MC3T3-E1, the mRNA and protein expression of Runt-related transcription factor 2, β-catenin, and bone morphogenetic protein 2 in MC3T3-E1 cells were significantly downregulated (P < 0.000 1), and the apoptosis rate significantly increased (P < 0.000 1), indicating that neutrophil extracellular traps can significantly inhibit osteogenic ability and promote apoptosis of MC3T3-E1 cells in vitro. After intervention with rutin alone or rutin combined with DNase I, the apoptosis and osteogenic ability of MC3T3-E1 cells under neutrophil extracellular traps were significantly improved, and the effect of rutin combined with DNase I was more significant than rutin alone, indicating that rutin may inhibit neutrophil extracellular trap formation, thereby improving the osteogenic ability of MC3T3-E1 cells. (3) Molecular docking and molecular dynamics simulations showed that rutin binds well to protein arginine deiminase 4, myeloperoxidase, and neutrophil elastase target proteins, indicating that rutin can target and inhibit neutrophil extracellular trap formation.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21344
BACKGROUND: Neural stem cells located in the ventricular zone and subventricular zone are crucial for cortical neurodevelopment and the treatment of neurodegenerative diseases. However, their precise regulatory mechanisms remain incompletely understood. miRNA-9 is one of the most abundantly expressed miRNAs in the vertebrate embryonic and adult brain, playing diverse roles during development. Nevertheless, the role of miR-9 in neural stem cell differentiation remains unclear. OBJECTIVE: To investigate the role of miR-9 in regulating the differentiation of neural stem cells in the ventricular zone and subventricular zone. METHODS: Neural stem cells were isolated from the ventricular zone and subventricular zone of embryonic day 14.5 ICR mice and cultured in proliferation medium for 3-4 days to form neurospheres. Stemness was identified by Pax6/Nestin immunofluorescence double staining. The expression profile of miR-9 was detected by qRT-PCR in telencephalon tissues at embryonic days 12.5, 14.5, 16.5, 18.5 and postnatal days 0, 7, as well as in embryonic day 14.5 neural stem cells cultured in vitro. Neural stem cells were transfected with miR-9 inhibitor or mimic using transfection reagents. After 24 hours, cells were differentiated for 3-4 days (neurons) and 6-8 days (glial cells). The differentiation of each lineage was quantified by immunofluorescence staining for Tuj1 (neuronal marker), myelin basic protein (oligodendrocyte marker), and glial fibrillary acidic protein (astrocyte marker). RESULTS AND CONCLUSION: qRT-PCR results showed that miR-9 was highly expressed in early embryonic telencephalon (E12.5-E14.5) and gradually decreased with development (E16.5 to P7). In E14.5 neural stem cells, miR-9 expression level was close to 90% of the internal reference RNU6B. Functional experiments showed that compared with the control group, the miR-9 inhibition group had decreased proportions of Tuj1-positive neurons and myelin basic protein-positive oligodendrocytes, while the proportion of glial fibrillary acidic protein-positive astrocytes increased. Conversely, the miR-9 overexpression group had increased proportions of Tuj1-positive neurons and myelin basic protein-positive oligodendrocytes, and decreased proportion of glial fibrillary acidic protein-positive astrocytes, with significant differences (P < 0.001). These results indicate that miR-9 plays a bidirectional regulatory role in neural stem cell differentiation: (1) It participates in the temporal regulation of neurogenesis through developmental stage-specific expression patterns (high early, downregulated later); (2) It maintains the balance of trilineage differentiation by promoting neuronal and oligodendrocyte differentiation while inhibiting astrocyte generation.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21340
BACKGROUND: Allogeneic hematopoietic stem cell transplantation may be complicated by central nervous system diseases not related to primary disease infiltration. There is no clear conclusion on the clinical symptoms, possible causes, and prognosis. OBJECTIVE: To explore the clinical characteristics, risk factors, and prognosis of non-primary infiltration-related central nervous system diseases after allogeneic hematopoietic stem cell transplantation, in order to provide evidence-based basis for early clinical diagnosis, etiological intervention, and prognosis improvement. METHODS: A retrospective analysis was conducted on clinical data, laboratory characteristics, and treatment processes of 298 patients with hematological diseases who developed non-primary infiltration-related central nervous system diseases after allogeneic hematopoietic stem cell transplantation from January 2015 to June 2024. They were divided into a non-primary infiltration-related central nervous system disease group (n=19) and a control group (without such diseases, n=279). Risk factors were analyzed statistically, and clinical symptoms, possible causes, and prognosis were evaluated. RESULTS AND CONCLUSION: (1) Among 298 patients, 19 developed non-primary infiltration-related central nervous system diseases, with an incidence of 6.4%. (2) The median onset time was 16 days (2-45 days) after transplantation. The main initial symptom was convulsions, accompanied by elevated blood pressure, headache, visual decline, consciousness disorders, and psychiatric behavioral abnormalities. (3) Univariate analysis showed significant associations between the occurrence of these diseases and granulocyte engraftment time, platelet engraftment time, history of central nervous system leukemia before transplantation, and grade III-IV graft-versus-host disease. Etiological analysis revealed: calcineurin inhibitor-related encephalopathy (2 cases), central nervous system injury (4 cases), central nervous system infection (4 cases), transplantation-associated thrombotic microangiopathy (4 cases), central nervous system graft-versus-host disease (1 case), intracranial hemorrhage (2 cases), endocrine metabolic encephalopathy (1 case), and unknown cause (1 case). (4) As of the follow-up date, the cumulative mortality in the disease group was 47% (9/19), significantly higher than 28.6% (80/279) in the control group. Further analysis showed that the estimated overall survival rates at 1 and 2 years after transplantation were significantly lower in the disease group than in the control group. In conclusion, non-primary infiltration-related central nervous system diseases after allogeneic hematopoietic stem cell transplantation are caused by multiple transplantation-related factors. Timely identification of pathogenic factors and precise diagnosis and treatment are crucial for improving the prognosis of patients with these complications.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21438
BACKGROUND: Early diagnosis and treatment of osteoarthritis remain a significant challenge due to the lack of highly specific biomarkers. OBJECTIVE: To screen characteristic genes of osteoarthritis, predict potential food-medicine homology traditional Chinese medicine and their core components, and validate their therapeutic potential through molecular docking and molecular dynamics simulations. METHODS: This study is based on three datasets (GSE55235, GSE169077, and GSE55457) from the GEO database, including a total of 25 normal samples and 26 osteoarthritis samples. It combines genes extracted from the eQTL database as exposure factors, and osteoarthritis data from the IEU openGWAS database (407,746 samples) as outcome factors. Core biomarkers were identified using least absolute shrinkage and selection operator regression, random forest, and support vector machine algorithms. CIBERSORT was used to evaluate immune infiltration characteristics and single-gene gene set enrichment analysis was performed. Potential traditional Chinese medicines were predicted using Coremine Medical and HERB databases, and food-medicine homology traditional Chinese medicines and their core components were screened, followed by molecular docking and molecular dynamics simulations. RESULTS AND CONCLUSION: ① Two genes, glucose transporter 3 (GLUT3) and atypical chemokine receptor 1 (ACKR1), were identified as characteristic genes of osteoarthritis, showing good diagnostic efficacy (AUC > 0.8) and involvement in metabolic regulation, cell signal transduction, and inflammatory responses, closely related to glucose metabolism, immune regulation, and inflammatory signaling pathways. ② Seven food-medicine homology traditional Chinese medicines were screened, including Cornus officinalis, Perilla frutescens, Ganoderma lucidum, Gastrodia elata, bitter almond, clove, and Rehmannia glutinosa, with core components β-sitosterol and stigmasterol. Molecular docking and dynamics simulations showed that stigmasterol had the best affinity with GLUT3 and the complex exhibited high stability. ③ This study systematically reveals the key roles of GLUT3 and ACKR1 in the pathogenesis of osteoarthritis, preliminarily validates the possibility of food-medicine homology traditional Chinese medicines intervening in the pathological process of osteoarthritis through multi-target and multi-pathway mechanisms, providing new molecular evidence for early diagnosis and targeted therapy, as well as theoretical support for prevention and treatment strategies. This research, from the perspective of traditional Chinese medicine and modern molecular biology, can provide a reference for the clinical application of traditional Chinese medicine in osteoarthritis.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21433
Background: Vestibular peripheral vertigo is characterized by a wide range of diseases, profound impact, and difficulty in prevention and treatment, posing a significant public health issue globally. Currently, randomized controlled trials (RCTs) of traditional Chinese medicine (TCM) for vestibular peripheral vertigo are complex and diverse, lacking standardized and systematic categorization, which constrains the quality of evidence-based evidence and clinical translation value. Objective: To systematically review RCTs of TCM for vestibular peripheral vertigo, analyze clinical research characteristics and outcome measures, to optimize the development of clinical guideline indicator sets and provide reference for future clinical trial design. Methods: PubMed, Web of Science, The Cochrane Library, EMbase, CNKI, VIP, Wanfang, and China Biology Medicine disc were searched from inception to April 18, 2025, to collect RCTs of TCM for vestibular peripheral vertigo. Two researchers independently conducted literature screening, data extraction, and quality assessment. Qualitative analysis was used to summarize clinical outcome measures and related trial design elements. Results and Conclusion: A total of 166 RCTs involving 14,718 patients were included, with 119 types of outcome measures. Outcome measures were categorized into 7 domains based on functional attributes: symptoms/signs, TCM syndromes, physical and chemical examinations, safety, long-term prognosis, quality of life, and others. Currently, the design of RCTs of TCM for vestibular peripheral vertigo has not formed a unified standard, with lack of standardization in TCM syndrome types and measurement tools, and numerous methodological quality issues. Clinical outcome measures exhibit prominent heterogeneity, imbalance between endpoint and surrogate indicators, unreasonable selection, neglect of economic indicators, and incomplete safety event and long-term prognosis evaluation systems. It is recommended that future researchers improve methodological quality, rationally design outcome measures, and align with the characteristics of TCM clinical trial efficacy evaluation, to promote the standardization of clinical guidelines and core indicator sets for vestibular peripheral vertigo, and provide more scientific and effective evidence-based medicine evidence for precise prevention and treatment.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21474
BACKGROUND: The tendon suturing technique for the hand has been continuously innovated with the development of biomechanics, minimally invasive techniques, and regenerative medicine. Over the past two decades, research has focused on optimizing traditional suturing techniques and the application of new repair materials, improving the effectiveness of tendon repair and the level of functional recovery in the hand. OBJECTIVE: To assess the global research status and development trends of hand tendon repair techniques over the past two decades through bibliometric analysis, identify research hotspots and their evolution. METHODS: Relevant literature was selected from the Web of Science database from 2005 to 2024, and bibliometric methods were employed for analysis. Data were organized using Microsoft Excel and analyzed for publication trends using the R language Bibliometrix package. VOSviewer was used to visualize keyword co-occurrence and collaboration networks, while CiteSpace was utilized to identify research hotspots and their temporal evolution. RESULTS AND CONCLUSION: Over the past two decades, research in the field of tendon suturing has shown a fluctuating growth trend. The United States, China, and Europe are the main contributing countries, with the United States occupying a central position in the global research network. Research on flexor tendon repair mainly focuses on biomechanics and the development of new repair materials, while extensor tendon research emphasizes postoperative functional recovery and complex injury repair. In recent years, biomaterials and regenerative medicine have gradually become research hotspots, promoting the application of precision medicine in tendon repair. In the future, interdisciplinary collaboration and the combination of advanced materials will further optimize hand tendon repair techniques.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21377
BACKGROUND: Conventional percutaneous CT-guided interventional puncture cannot be monitored in real time, and the operation takes a long time. In some high-risk puncture sites, multiple CT scans and adjustments to the position of the puncture needle are required, causing greater radiation damage to the patient. A self-developed surgical navigation and positioning system provides an effective way to solve the clinical problems of percutaneous puncture information perception and accurate and safe target puncture in complex intraoperative environments, achieving precise puncture positioning of the chest and abdomen. OBJECTIVE: To evaluate the safety, effectiveness and usability of the self-developed percutaneous puncture navigation robotic system in clinical application. METHODS: A retrospective analysis was conducted on clinical trial data from percutaneous lung nodule biopsy and tumor ablation procedures guided by a puncture navigation robotic system at the First Affiliated Hospital of Guangzhou Medical University and the Second Affiliated Hospital of Soochow University between November 1, 2021 and June 28, 2022. A multicenter, open-label, parallel controlled clinical study was conducted, and 120 subjects were randomly divided into an experimental and a control group, with 60 subjects in each group. The experimental group underwent puncture guided by a puncture navigation robotic system, while the control group underwent conventional CT-guided percutaneous puncture. The primary effectiveness endpoint was puncture accuracy rate, and secondary endpoints included number of needle adjustments, one-time success rate of puncture, number of CT scans, and system usability. RESULTS AND CONCLUSION: No unsafe events occurred during the entire clinical trial. The one-time success rate of puncture was 98.31% in the experimental group and 15.00% in the control group; complication rates were 6.78% and 13.33%, respectively. The system usability satisfaction rate was 100%. The system achieved interactive modeling of puncture needle and soft tissue, dynamic reconstruction of complex operative environment and real-time perception of puncture information, and dynamic navigation and tracking compensation under physiological motion and puncture interaction, providing an effective solution to the clinical challenges of information perception and accurate and safe target puncture in complex intraoperative environments.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21444
BACKGROUND: Icariin has the dual activity of promoting bone formation and inhibiting bone resorption, but its clinical application is plagued by low bioavailability, difficulty in controlling dosage, and a high risk of adverse reactions. OBJECTIVE: To prepare a three-dimensional scaffold containing icariin sustained-release microspheres and characterize their osteogenic activity in vitro. METHODS: A silk fibroin/chitosan/nanohydroxyapatite three-dimensional scaffold (SF/CS/nHA scaffold), icariin sustained-release microspheres, and a silk fibroin/chitosan/nanohydroxyapatite three-dimensional scaffold loaded with icariin sustained-release microspheres (SF/CS/nHA-ICA scaffold) were prepared. The drug loading efficiency, encapsulation efficiency, and in vitro drug release of the icariin sustained-release microspheres were characterized. The pore size, porosity, water absorption expansion rate, and hot water dissolution rate of the two scaffolds were measured. Rabbit bone marrow mesenchymal stem cells (or human rheumatoid arthritis fibroblast-like synoviocytes) were seeded on SF/CS/nHA and SF/CS/nHA-ICA scaffolds, with cells cultured alone as controls. Cell adhesion was observed by scanning electron microscopy. Cell proliferation and viability were assessed by CCK-8 assay, live/dead staining, and F-actin staining. The mRNA and protein expression of Runx-2, osteocalcin, and type I collagen in bone marrow mesenchymal stem cells were detected by RT-qPCR and western blot. RESULTS AND CONCLUSION: (1) The drug loading efficiency and encapsulation efficiency of icariin sustained-release microspheres were (29.38±0.04)% and (52.01±0.09)%, respectively, and the microspheres could sustainably release icariin for more than 90 days in vitro. (2) Scanning electron microscopy showed a honeycomb-like porous structure with interconnected pores in both scaffolds. There were no significant differences in pore size, porosity, water absorption expansion rate, or total hot water dissolution rate between the two groups (P > 0.05). (3) Scanning electron microscopy showed that both cell types adhered tightly to the scaffold surface and pores, with more extended pseudopodia on the SF/CS/nHA-ICA scaffold. CCK-8 assay, live/dead staining, and F-actin staining showed that compared with the control and SF/CS/nHA groups, the SF/CS/nHA-ICA scaffold promoted the proliferation and viability of rabbit bone marrow mesenchymal stem cells, while inhibiting the proliferation and viability of human rheumatoid arthritis fibroblast-like synoviocytes. (4) RT-qPCR and western blot showed that compared with the control and SF/CS/nHA groups, the mRNA and protein expression of Runx-2, osteocalcin, and type I collagen were increased in the SF/CS/nHA-ICA group (P < 0.05). (5) These results indicate that the icariin sustained-release microsphere three-dimensional scaffold has good cytocompatibility, and in vitro osteogenic and anti-inflammatory effects.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21489
BACKGROUND: The pathological process of Alzheimer's disease is closely related to β-amyloid/Tau deposition and cerebrovascular dysfunction, and existing therapies are difficult to effectively intervene. In recent years, research on glucagon-like peptide-1 receptor agonists for the treatment of Alzheimer's disease has gained increasing popularity. As a novel dual agonist of the glucagon-like peptide-1 receptor and glucagon receptor, the therapeutic mechanism of Mazdutide in Alzheimer’s disease remains to be elucidated.
OBJECTIVE: To systematically explore the mechanism by which Mazdutide improves cognitive function in Alzheimer's disease using network pharmacology and experimental validation, and to identify potential core molecular targets, providing a theoretical basis for new therapeutic strategies.
METHODS: A multi-omics integration strategy was employed. Co-localization targets of Alzheimer's disease and Mazdutide were screened using DisGeNET and SEA databases. A protein-protein interaction network was constructed via STRING, and hub genes were identified by Cytoscape topological analysis. In APP/PS1/Tau triple transgenic Alzheimer's disease mice, Mazdutide (30 nmol/kg) was administered intraperitoneally. Cognitive function was assessed using Morris water maze, novel object recognition, and Y maze tests. Western blot was used to detect hippocampal β-amyloid (6E10), p-Tau181, and endothelin receptor A expression.
RESULTS AND CONCLUSION: ① Fifty-two co-localized targets were identified, with endothelin receptor A (EDNRA) as the top hub gene. ② Mazdutide significantly improved cognition in 3xTg mice: shortened spatial memory latency, increased novel object recognition index, increased spontaneous alternation in Y maze, and reduced hippocampal pathological burden, with decreased p-Tau181 and EDNRA overexpression. ③ Gene Ontology and KEGG enrichment revealed core pathways: biological processes included blood pressure regulation, amine transport regulation, and amine transport; cellular components included symmetric synapses, sperm head, and pseudopodia; molecular functions included G protein-coupled peptide receptor activity, peptide receptor activity, and neuropeptide receptor activity. KEGG pathways included neuroactive ligand-receptor interaction, calcium signaling, and hormone signaling. ④ This study confirms that Mazdutide may improve cognitive deficits in Alzheimer's disease by targeting and inhibiting EDNRA overexpression, reducing neuronal pathological damage, and acting through multiple dimensions. The discovery of EDNRA provides a novel therapeutic target for Alzheimer's disease.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21624
BACKGROUND: Lower extremity deep vein thrombosis is a catastrophic complication after lumbar spine fusion, and previous studies have shown that some patients, such as those with prosthetic arthroplasty and abdominal surgeries, suffer from a combination of preoperative anemia, which predisposes them to postoperative deep vein thrombosis of the lower extremities. However, whether preoperative anemia increases the risk of lower extremity deep vein thrombosis after lumbar spine fusion is unclear. OBJECTIVE: To investigate the correlation between preoperative anemia and deep vein thrombosis of the lower extremities in patients after lumbar fusion. METHODS: The clinical data of 1 178 patients who underwent lumbar spinal fusion treatment admitted to Third Affiliated Hospital of Anhui Medical University from January 2020 to December 2023 were retrospectively analyzed. According to whether or not the patients developed lower extremity deep vein thrombosis after the operation, the patients were divided into the deep vein thrombosis group (ultrasound report suggestive of lower extremity deep vein thrombosis) and the non-deep vein thrombosis group (ultrasound report suggestive of no lower extremity deep vein thrombosis). The incidence of anemia was compared between the two groups, and risk factors for lower extremity deep vein thrombosis in lumbar fusion patients were determined by univariate analysis and multivariate logistic regression analysis. RESULTS AND CONCLUSION: Among 1 178 patients, there were 43 cases (3.7%) in the deep vein thrombosis group and 1 135 cases (96.4%) in the non-deep vein thrombosis group. The incidence of preoperative anemia in the deep vein thrombosis group was 32.6%, significantly higher than that in the non-deep vein thrombosis group (10.9%, P < 0.05). Univariate analysis showed significant differences between the two groups in preoperative hemoglobin (P < 0.001), preoperative red blood cell count (P=0.028), D-dimer positivity (P=0.029), hypertension (P=0.019), number of fused segments (P=0.023), anemia (P < 0.001), and blood transfusion (P=0.006) (P < 0.05). Multivariate analysis showed that preoperative anemia (OR=4.221, 95%CI: 1.198-14.802, P=0.025) and D-dimer positivity (OR=2.023, 95%CI: 1.065-3.844, P=0.031) were risk factors for lower extremity deep vein thrombosis after lumbar fusion. These findings suggest that the incidence of preoperative anemia is relatively high in patients undergoing elective lumbar fusion, and preoperative anemia is an independent risk factor for deep vein thrombosis after lumbar fusion. It is recommended that preoperative anemia should be actively managed and corrected before elective lumbar fusion to reduce the risk of lower extremity deep vein thrombosis.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21614
BACKGROUND: Meniscus injury, as a significant contributing factor to knee joint degeneration, can accelerate the progression of osteoarthritis through anterior horn tears that alter joint stress distribution. Existing studies primarily focus on biomechanical changes in normal bone conditions, while the regulatory role of different bone conditions on the injury mechanism remains unclear. OBJECTIVE: To investigate the effect of medial meniscus anterior horn transverse tears on the biomechanical differences of knee joints with varying bone conditions. METHODS: Imaging data of the lower limb from a healthy adult were used to construct a normal knee joint model in Mimics 2017. The model was further optimized and assembled using Geomagic Studio 2017 and SolidWorks 2017, and a medial meniscus anterior horn transverse tear model was established. Material properties were assigned in Ansys Workbench 2017, and three finite element models were created to simulate normal bone, reduced bone mass, and osteoporosis conditions. The models were validated using anterior drawer and axial loading tests. With the femur constrained and the distal tibia fixed, a 1000 N axial compressive load was applied to calculate stress peaks and strain distributions. RESULTS AND CONCLUSION: (1) Under static standing conditions, both normal and injured medial meniscus models showed significantly higher joint stress loads in reduced bone mass and osteoporosis groups compared to normal bone mass group. (2) The medial meniscus anterior horn transverse tear model exhibited a progressive increase in stress loads on femoral cartilage and both menisci compared to the normal model, while stress on tibial cartilage surfaces decreased with bone mass reduction. In the meniscus injury model, stress concentrated at the tear edges. (3) Subchondral bone regions showed elevated stress levels under reduced bone mass conditions, especially after medial meniscus injury, with significantly increased strain distribution range and equivalent stress peaks (P < 0.05). (4) These findings suggest that medial meniscus anterior horn transverse tears under reduced bone density exacerbate joint contact area reduction and local stress concentration, potentially leading to increased local strain in tibial subchondral bone, indicating an interactive biomechanical effect between bone degeneration and meniscus injury. This provides a reference for meniscus repair strategies considering bone quality differences.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21550
BACKGROUND: Genetic factors play an important role in the pathophysiology of osteoporosis, and Mendelian randomization can be used to infer causal associations between specific genes and diseases using eQTLs. OBJECTIVE: To identify potential therapeutic targets for osteoporosis based on druggable genes-related Mendelian randomization and colocalization analysis, to explore the potential biological mechanisms in the treatment of osteoporosis using bioinformatics analysis, and to predict the binding activity of drug targets using drug enrichment analysis and molecular docking. METHODS: (1) Data sources: Druggable genes were sourced from the DGIdb database (a public database constructed by the University of Washington School of Medicine, widely used for drug target discovery) and information provided in the literature. Expression quantitative trait locus (eQTL) data for druggable genes were obtained from eQTLGen (a large-scale blood eQTL database jointly constructed by multiple international research institutions, including the University of Groningen). Osteoporosis genome-wide association study (GWAS) data were obtained from FinnGen R12 (a large genomic database led by the University of Helsinki), including 10,461 osteoporosis cases and 473,264 controls. GEO datasets GSE230665 (microarray) and GSE169396 (single-cell) were also used. (2) Methods: Genes closely related to osteoporosis were screened; expression of genes in osteoporosis was evaluated via microarray data; single-cell analysis further observed the regulatory role of genes in cell communication; enrichment analysis was used to elucidate biological functions, and protein-protein interaction networks were constructed to analyze potential associations; drug enrichment and molecular docking predicted and simulated the binding of small molecule drugs to targets. RESULTS AND CONCLUSION: The study identified 37 druggable genes associated with osteoporosis, among which troponin C2 and CXC chemokine receptor 6 (CXCR6) had protective effects and shared causal genetic variants with the disease. Microarray data analysis showed that CXCR6 expression was significantly lower in osteoporosis than in normal controls, suggesting a weakened protective effect. Single-cell analysis further revealed that CXCR6 was mainly expressed on T cells, and CXCR6+ T cells exhibited stronger cell communication capabilities. Drug enrichment analysis found that NSC95397 could target CXCR6, and molecular docking showed good binding activity. These findings not only provide clues for the development of new drugs for osteoporosis but also facilitate the translation of research results.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21586
BACKGROUND: In recent years, hydrogels have become an important research direction in the treatment of rheumatoid arthritis due to their excellent biocompatibility, controllable drug release performance, and advantages in multiple drug delivery routes. OBJECTIVE: To systematically review the application of hydrogel-based materials as drug delivery carriers in the treatment of rheumatoid arthritis, and explore the impact of different administration routes on the therapeutic effect. METHODS: Using “hydrogel, rheumatoid arthritis, smart hydrogel system, injectable hydrogel, intra-articular injection, transdermal drug delivery” as Chinese and English search terms, we searched PubMed, Web of Science, CNKI, WanFang Data, and VIP. Based on the inclusion criteria, 62 articles were finally included for review. RESULTS AND CONCLUSION: Hydrogels, leveraging their three-dimensional network structures and tunable physicochemical properties, not only allows drugs to accurately reach the lesion area but also significantly prolongs the retention time of drugs in the joint cavity, making them an ideal carrier in the field of drug delivery. The drug release mechanisms of hydrogels mainly include diffusion, chemical regulation, and swelling-mediated release; in addition, stimulus-responsive hydrogels can dynamically regulate drug release behavior based on environmental conditions (such as pH, temperature, enzyme concentration, etc.). In the treatment of rheumatoid arthritis, common administration routes for hydrogel drug delivery systems include parenteral administration, oral administration, transdermal administration, and intra-articular injection, which significantly reduce systemic adverse reactions, improve drug absorption efficiency, and enhance patient compliance. Although hydrogels as drug delivery carriers have shown significant application potential in the treatment of rheumatoid arthritis, long-term safety, biodegradability, and large-scale production still need breakthroughs to promote the clinical translation of hydrogel drug delivery carriers.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21572
BACKGROUND: Hydroxyapatite is widely used in bone tissue engineering due to its excellent osteoconductivity. However, its limited osteoinductivity restricts its clinical application and therapeutic efficacy. OBJECTIVE: To construct manganese-doped hydroxyapatite/polydopamine composite bone graft materials and characterize their physicochemical and biological properties. METHODS: (1) Using Ca(NO₃)₂·4H₂O as the calcium source, (NH4)2HPO4 as the phosphorus source, and manganese nitrate solution as the manganese source, manganese-doped hydroxyapatite was prepared by hydrothermal homogeneous coprecipitation with manganese/(calcium+manganese) molar ratios of 5%, 10%, and 15%, respectively. The corresponding materials were denoted as 5Mn-HA, 10Mn-HA, and 15Mn-HA. Hydroxyapatite and the three manganese-doped hydroxyapatite materials were immersed in dopamine hydrochloride-Tris buffer solution to prepare manganese-doped hydroxyapatite/polydopamine composites, denoted as HA/PDA, 5Mn-HA/PDA, 10Mn-HA/PDA, and 15Mn-HA/PDA. The morphology, manganese ion release, degradation rate, and cytocompatibility were characterized. The manganese-doped material with better cytocompatibility was selected for subsequent experiments. (2) Hydroxyapatite, HA/PDA, 5Mn-HA, and 5Mn-HA/PDA were co-cultured with rat bone marrow mesenchymal stem cells. After osteogenic induction, alkaline phosphatase activity and alizarin red S staining were performed, and q-PCR was used to detect Runx2 and osteocalcin mRNA expression. (3) In 24 SD rats, two circular full-thickness bone defects of 5 mm diameter were created on each side of the calvarium. The right defects were implanted with hydroxyapatite, HA/PDA, 5Mn-HA, or 5Mn-HA/PDA (6 rats per material), while the left defects served as blank controls. At 4 and 8 weeks post-surgery, samples were harvested for Micro-CT scanning, hematoxylin-eosin and Masson staining. RESULTS AND CONCLUSION: (1) The manganese-doped hydroxyapatite/polydopamine composites were microspherical with particle sizes ranging from 9.86 to 13 μm. With increasing manganese doping, the release of manganese ions increased, and the release rate from the composites was lower than that from the corresponding manganese-doped hydroxyapatite. The composites exhibited faster degradation rates compared to manganese-doped hydroxyapatite. Based on live/dead staining and CCK-8 assays, 5Mn-HA showed no obvious cytotoxicity; therefore, 5Mn-HA and 5Mn-HA/PDA were selected for further osteogenic evaluation and in vivo verification. (2) Combined results of alkaline phosphatase activity, alizarin red S staining, and q-PCR indicated that 5Mn-HA/PDA had the strongest osteogenic ability among the four materials. (3) Micro-CT scanning showed that the blank control group had the slowest bone repair and least new bone formation, while the 5Mn-HA/PDA group had the fastest bone repair and most new bone. Hematoxylin-eosin and Masson staining further confirmed the Micro-CT results. (4) In conclusion, manganese-doped hydroxyapatite/polydopamine composites possess good physicochemical and biological properties.
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.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21598
BACKGROUND: In recent years, the involvement of non-apoptotic regulated cell death in the development of ischemic stroke has become a research hotspot. OBJECTIVE: To summarize the roles and action mechanisms of non-apoptotic regulated cell death subroutines such as autophagy, ferroptosis, cuproptosis, disulfidptosis, necroptosis, and pyroptosis in the neuronal damage caused by ischemic stroke. METHODS: Relevant literature on non-apoptotic regulated cell death and ischemic stroke was retrieved from the China National Knowledge Infrastructure and PubMed databases. The search terms included "ischemic stroke, regulated cell death, autophagy, ferroptosis, cuproptosis, disulfidptosis, necroptosis, pyroptosis, alkaliptosis, oxeiptosis, parthanatos, mitochondrial permeability transition-driven necrosis, neutrophil extracellular trap-related death, lysosome-dependent cell death, immunogenic cell death, anoikis" in English and corresponding Chinese terms. Based on inclusion criteria, 176 articles were finally included for analysis and summary. RESULTS AND CONCLUSION: The regulatory mechanisms of non-apoptotic regulated cell death mainly include autophagy, ferroptosis, anoikis, cuproptosis, disulfidptosis, necroptosis, pyroptosis, oxeiptosis, alkaliptosis, parthanatos, mitochondrial permeability transition-driven necrosis, neutrophil extracellular trap-related death, lysosome-dependent cell death, and immunogenic cell death. Autophagy plays a dual regulatory role in neuronal injury after ischemic stroke: under ischemic conditions, autophagy exerts a neuroprotective effect, whereas excessive autophagy during reperfusion can lead to neuronal death. Ferroptosis can aggravate neuronal injury in ischemic stroke through iron overload and lipid peroxidation. Cuproptosis can regulate glutathione-induced ferroptosis by modulating the protein ferredoxin 1. There is partial crosstalk between disulfidptosis and ferroptosis; under glucose deprivation, upregulation of solute carrier family 7 member 11 consumes NADPH, leading to abnormal accumulation of disulfide compounds and promoting disulfidptosis in neurons. Mixed lineage kinase domain-like pseudokinase, a key participant in necroptosis, is also associated with activation of the pyroptosis-related protein NLRP3 inflammasome, further promoting neuronal pyroptosis during necroptosis in ischemic stroke. Neutrophil extracellular trap-related death in ischemic stroke is mainly caused by citrullination, stress-triggered neutrophil extracellular trap formation, and inflammatory responses mediated by release of various cytotoxic proteases. Other emerging subtypes such as immunogenic cell death cause neuronal damage in ischemic stroke through various specific mechanisms.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21585
BACKGROUND: Previous studies have demonstrated neuroprotective potential of microRNA-23a-3p in traumatic brain injury. However, direct evidence is still lacking regarding whether this protective effect stems from its precise regulation of the M1/M2 polarization balance of microglia. OBJECTIVE: To clarify the expression changes of microRNA-23a-3p in mouse brain tissue after traumatic brain injury and to explore the specific mechanism by which it affects neurological function through regulating microglial polarization. METHODS: Eighty C57BL/6J mice were randomly assigned to four groups: a sham operation group, a traumatic brain injury group, a traumatic brain injury + agomir-NC group, and a traumatic brain injury + agomir-MicroRNA-23a-3p group. The traumatic brain injury model was established using the cortical impact method. The sham group did not receive cortical impact. The intervention groups received intracerebroventricular injection of agomir-NC or agomir-MicroRNA-23a-3p after modeling. Six mice from the sham and traumatic brain injury groups were analyzed at 1, 3, 7, and 14 days post-injury, and six mice from the other two groups were analyzed at 14 days post-injury. Neurological deficits were assessed using the modified neurological severity score (mNSS). Hematoxylin-eosin staining and Nissl staining were used to observe pathological changes in brain tissue and neurons. qRT-PCR and western blot were used to detect the expression levels of MicroRNA-23a-3p, M1 markers (CD16, CD86), M2 markers (CD206, arginase-1), and inflammatory cytokines (tumor necrosis factor-α and interleukin-10). Immunohistochemistry was used to evaluate microglial M1/M2 polarization and the aggregation of F4/80-positive cells in the injured area. RESULTS AND CONCLUSION: Compared with the sham group, the expression of MicroRNA-23a-3p in the traumatic brain injury group showed a "V"-shaped curve, with downregulation in the early phase and upregulation starting at 7 days post-injury. Upregulation of MicroRNA-23a-3p reduced the mNSS score in traumatic brain injury mice. Morphological results showed that upregulation of MicroRNA-23a-3p alleviated brain edema and neuronal damage. Molecular biology results showed that upregulation of MicroRNA-23a-3p promoted microglial polarization from M1 to M2 phenotype. These findings indicate that MicroRNA-23a-3p can promote neurological function recovery after traumatic brain injury in mice by regulating microglial polarization.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21581
BACKGROUND: In recent years, magnetic stimulation therapy can activate the classical transient receptor potential channel 1, triggering the calcium-mitochondrial axis to enhance myogenesis and mitochondrial biogenesis in vivo, thereby recapitulating physiological adaptations related to exercise-induced metabolic responses. As an emerging technique for promoting muscle function, magnetic stimulation has gained widespread attention and validation in the rehabilitation of muscular diseases due to its advantages of being non-invasive, passive, and safe. However, there is a lack of clinical studies on the therapeutic efficacy of this technique in the treatment of disuse-induced muscle atrophy. OBJECTIVE: To investigate the therapeutic effect of exercise therapy combined with magnetic stimulation on the recovery of muscle strength and locomotor ability in patients with disuse-induced muscle atrophy of the lower limbs. METHODS: Sixteen patients with lower limb disuse muscle atrophy caused by prolonged bed rest after unilateral Achilles tendon rupture surgery were recruited and randomly divided into control group and experimental group, 8 cases in each group. The control group received traditional exercise rehabilitation therapy, including joint range of motion training, muscle strength training, and soft tissue stretching training, 3 times a week. The experimental group additionally received medical magnetic physical factor stimulation (intensity 1.5 mT, frequency 3 300 Hz, 48 h per session, 10 min each time) on this basis, with a total trial duration of 4 weeks. All subjects underwent maximum voluntary contraction (MVC) test of the lower limbs and gait speed measurements including Timed Up and Go test (TUG), 5-times sit-to-stand test (5STS), and 6 m normal walking speed test before and after intervention. RESULTS AND CONCLUSION: After 4 weeks of intervention, all 16 subjects completed the trial. In the experimental group, the maximum voluntary contraction of the affected lower limb (P=0.001) and the difference rate of MVC between affected and healthy sides (P=0.001) significantly decreased, and the improvements were superior to those in the control group. In terms of gait speed indicators, the experimental group showed significant improvements in TUG (P=0.038), 6 m normal walking speed (P=0.025), and 5STS (P=0.050) compared with baseline. Between-group comparison revealed that the experimental group had significantly greater improvements in MVC of the affected leg (P=0.003), difference rate of MVC between affected and healthy sides (P=0.004), TUG (P=0.019), and 6 m normal walking speed (P=0.011) than the control group. These data confirm that after 4 weeks of low-frequency pulsed magnetic field (1.5 mT, 3 300 Hz) combined with exercise therapy, patients with disuse muscle atrophy after Achilles tendon rupture showed significantly better improvements in MVC of the affected and healthy legs, TUG, and 6 m normal walking speed than the control group, demonstrating that magnetic stimulation combined with exercise therapy has an auxiliary synergistic effect on isometric muscle strength and lower limb motor function. Therefore, magnetic stimulation combined with exercise therapy can be used as a new means for rehabilitation of disuse muscle atrophy.
Stem Cell Research & Therapy•2026•DOI: 10.1186/s13287-026-05027-z
Retinal organoids (ROs) derived from human pluripotent stem cells are crucial for modeling retinal development and disease. However, the functional electrophysiological maturation of photoreceptors within ROs remains poorly characterized. This study aimed to define the functional maturation timeline of photoreceptors in human embryonic stem cell (hESC)-derived ROs. H9 hESC-derived ROs which included a CRX-tdTomato reporter line for specific photoreceptor identification were utilized. An integrated approach of RNA-sequencing analysis, immunofluorescence staining, and whole-cell patch-clamp recordings was employed to systematically assess photoreceptor maturation over 300 days of differentiation. Transcriptional and protein analysis revealed progressive upregulation of key ion channels. Patch-clamp recordings demonstrated stage-dependent maturation of membrane properties, which stabilized by D120–125. Hyperpolarization-activated cyclic nucleotide-gated (HCN) channel-mediated currents (Ih) increased progressively, peaking at D240, with amplitudes comparable to mature primate photoreceptors. Voltage-gated sodium (Nav) currents also showed significant developmental upregulation, reaching a maximum, stable plateau from D210–215 onward. Pharmacological blockade confirmed the identity of HCN and Nav currents. Critically, the capacity for action potential (AP) generation increased developmentally, with the proportion of photoreceptors capable of firing APs rising from 16.7% at D90–95 to a peak of 90.2% by D240–245. This study defines a comprehensive electrophysiological maturation timeline for photoreceptors in human ROs and establishes D240 as a key benchmark for functional maturity, characterized by peak Ih currents and AP generation capacity equivalent to mature native photoreceptors. These findings provide essential physiological criteria for standardizing RO quality control, enhancing their utility for modeling retinal degenerative diseases and developing cell replacement therapies.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2026046
Epstein-Barr virus-associated gastric cancer (EBVaGC) displays unique clinicopathological hallmarks, yet serology-based tools for its detection are still limited. Here, we develop a functional EBV proteome microarray covering 72 viral proteins and apply it to profile antibody responses in 62 gastric cancer patients. The resulting landscape reveals an IgG-skewed humoral signature specific to EBVaGC and identifies 34 EBV antigens exhibiting differential reactivities. Multivariable logistic regression integrates complementary markers into an optimal five-analyte panel (LF2_IgG, BBLF2_IgG, BLRF2_IgG, BPLF1-2_IgA, and BGLF4_IgA) that achieves outstanding discrimination performance (AUC = 0.93) in an independent validation set (n = 316). The panel’s performance is further validated in a community-based screening cohort (n = 474), where it achieves 87.3% sensitivity and 88.3% specificity for distinguishing EBVaGC from non-malignant gastric conditions spanning gastritis to dysplasia (AUC = 0.94). Together, these results establish a serological framework for EBVaGC diagnosis and provide a scalable strategy for population-level screening that could materially improve the management of this virus-driven malignancy.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2026078
Sodium-glucose cotransporter 2 inhibitors (SGLT2i) have demonstrated clinical benefits in heart failure with preserved ejection fraction (HFpEF), yet the underlying mechanisms remain poorly defined. Given that mitochondrial dysfunction represents a central feature of HFpEF pathophysiology, we investigate whether modulation of mitochondrial homeostasis contributes to the cardioprotective effects of dapagliflozin. Using a Dahl salt-sensitive rat model of HFpEF, we find that dapagliflozin markedly improves diastolic function and attenuates cardiac hypertrophy, fibrosis, and apoptosis. These beneficial effects are accompanied by significant restoration of mitochondrial structure and function. Consistently, in an in vitro HFpEF model, dapagliflozin enhances mitochondrial respiratory capacity in cardiomyocytes, indicating a direct mitochondrial regulatory effect. Mechanistically, integrative transcriptomic and experimental analyses identify the SIRT1/PGC-1α/Mitofusin-2 (Mfn-2) signaling axis as a critical pathway suppressed in HFpEF but reactivated following dapagliflozin treatment. Activation of this pathway promotes mitochondrial biogenesis and improves mitochondrial dynamics, thereby preserving cardiomyocyte homeostasis. Collectively, our findings reveal that dapagliflozin exerts cardioprotective effects in HFpEF by restoring mitochondrial homeostasis through the SIRT1/PGC-1α/Mfn-2 axis, providing mechanistic insight into SGLT2i-mediated benefits and highlighting mitochondrial regulation as a potential therapeutic strategy for HFpEF.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2026035
Pulmonary fibrosis (PF) is a life-threatening interstitial lung disease characterized by scarring and inflammation in lung tissues. Aberrant activation of the JAK/STAT and NF-κB signaling pathways is critical in initiating and sustaining the inflammatory processes that drive fibrotic progression. In this study, we identify a novel small-molecule compound, T4015, a 4-indolyl-2-phenylaminopyrimidine derivative, as a dual-pathway inhibitor targeting both JAK/STAT and NF-κB signaling. Dual-luciferase reporter assays demonstrate the potent inhibitory activity of T4015 against these pathways. T4015 effectively suppresses the phosphorylation of STAT3, JAK1, and TYK2 induced by IL-6 and IFN-β, while suppressing LPS-induced NF-κB activation in macrophages. Transcriptome sequencing and pathway enrichment analyses further confirm that T4015 downregulates multiple inflammation-related signaling cascades, including the JAK/STAT, NF-κB, TNF, IL-17, and Toll-like receptor pathways. In a mouse model of bleomycin-induced PF, T4015 treatment significantly improves survival, attenuates collagen deposition, and reduces the expression of pro-inflammatory and profibrotic markers such as IL-6, CCL2, and COL1. Molecular docking and target prediction analyses suggest that T4015 exhibits strong binding affinity for multiple kinases within the JAK/STAT and NF-κB networks, including JAK1, TYK2, JAK2, JAK3, RIPK1, IRAK1/4, TAB1, and ZAP70. Collectively, these results highlight T4015 as a promising therapeutic candidate for PF through its simultaneous inhibition of the JAK/STAT and NF-κB signaling pathways.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2025055
Thromboxane A2 (TXA2) is a labile eicosanoid with a half-life of 30 s, limiting direct quantification. Its stable urinary metabolite, 11-dehydrothromboxane B2 (11dH-TXB2), reflects in vivo TXA2 biosynthesis and aspirin-mediated COX-1 inhibition. Creatinine normalization is required to correct for urine concentration and renal function. This study establishes a quantitative liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for simultaneous determination of 11dH-TXB2 and creatinine in human urine. Sample pretreatment involves organic phase extraction, eliminating solid-phase extraction. The method was validated for linearity, accuracy, precision, and recovery. The linear range for 11dH-TXB2 was 0.1–50 ng/mL (r² = 0.99656) and for creatinine 10–5000 ng/mL (r² = 0.99950). Quality control accuracies ranged from 85.83% to 113.21%, with RSDs below 9.71%. Standard recoveries were 85–110%. The assay meets regulatory requirements for simultaneous quantification. This approach provides a reliable tool for monitoring aspirin response and investigating thromboxane-related pathologies.
Chinese Journal of New Drugs•2025•DOI: cast_zgxyzz_1236731785313317742
Background: Heart failure with recovered ejection fraction (HFrecEF) is a distinct phenotype with unclear clinical characteristics and prognosis. Methods: We prospectively enrolled 1,234 patients with heart failure and reduced ejection fraction (HFrEF) from January 2015 to December 2018. After optimal medical therapy, 312 patients (25.3%) achieved recovery of left ventricular ejection fraction (LVEF) to ≥50% and were classified as HFrecEF. Clinical characteristics, medication use, and outcomes were compared with those who remained HFrEF. The primary outcome was a composite of all-cause death and heart failure hospitalization. Results: Compared with HFrEF patients, HFrecEF patients were younger, more likely to be female, had a higher prevalence of hypertension and atrial fibrillation, and had a shorter duration of heart failure. They had lower baseline levels of NT-proBNP and smaller left ventricular dimensions. Over a median follow-up of 3.2 years, HFrecEF patients had a significantly lower risk of the primary outcome (adjusted HR 0.45, 95% CI 0.32-0.63, p<0.001). However, 23.4% of HFrecEF patients experienced deterioration of LVEF during follow-up, and these patients had a worse prognosis compared with those who maintained recovery. Independent predictors of LVEF deterioration included ischemic etiology, diabetes, and non-adherence to guideline-directed medical therapy. Conclusions: HFrecEF is associated with a better prognosis than HFrEF, but a substantial proportion of patients may experience LVEF deterioration. Continued optimization of medical therapy and close monitoring are essential for this population.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2025138
Lung adenocarcinoma (LUAD) remains the leading cause of cancer-related mortality worldwide, with a five-year survival rate of approximately 15%. Despite advances in targeted therapy, the limited repertoire of actionable mutations and the inevitable emergence of drug resistance necessitate the continuous discovery of novel therapeutic agents. This study investigates the antitumor efficacy of EOAI3402143, a small-molecule inhibitor, against LUAD and elucidates its mechanism of action. Using flow cytometry, transwell, colony formation assays, western blot, RT-qPCR, and RNA-seq, we demonstrate that EOAI3402143 promotes apoptosis and suppresses migration, invasion, and proliferation of LUAD cells. Mechanistically, EOAI3402143 inhibits NF-κB pathway activation and downregulates NR4A1 expression, thereby attenuating LUAD progression. In vivo experiments confirm superior therapeutic efficacy of EOAI3402143 in LUAD models. These findings position EOAI3402143 as a promising candidate for LUAD therapy, particularly for patients with limited targeted options or acquired resistance.
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.