Stem Cell Research & Therapy•2026•DOI: 10.1186/s13287-026-04995-6
Background Gastrointestinal diseases often involve cellular damage, degeneration or dysfunction in the tract, frequently requiring surgical interventions risking complications and lowered quality of life. Regenerative medicine holds great promise in improving patient care and providing novel treatment options for previously irreparable and untreatable tissues. Despite the clinical potential of intestinal organoids as a resource for regenerative cell therapy and bioengineering, the lack of clinical-grade cultures has hampered further development. Moreover, strategies to efficiently and reliably expand clinical-grade cultures at the scale required for application is limited. Methods A GMP-compliant protocol was developed to generate patient-derived colonic organoids from endoscopic biopsies. Clinical-grade colonic organoids cultured and expanded in Type-I collagen were compared to conventional Matrigel cultured organoids. To improve the culture-, cost-, and time-efficiency of culture expansion, several strategies were developed including organoid area-based passaging, one well plate culture, and the incorporation of Wnt activating peptide, PG-008. Conventional recombinant WNT3A culture was compared to the peptide PG-008 culture using single cell RNA sequencing. Results Clinical-grade collagen cultured organoids exhibited similar culture efficiency to Matrigel. Organoid establishment rate from 60 patients using the GMP-compliant protocol was 82%. The incorporation of PG-008 significantly enhanced organoid growth and stabilized patient-patient variability through intestinal stem cell (ISC) enrichment. Single cell RNA sequencing revealed that PG-008 resulted in remarkably pure culture consisting of ISCs
Stem Cell Research & Therapy•2026•DOI: 10.1186/s13287-026-04974-x
Background Variants in OTUD5 are associated with neurodevelopmental disorders (NDDs), yet the underlying molecular mechanisms remain unclear. This study aimed to investigate the pathogenicity of a novel OTUD5 variant (c.697G > A, p.Val233Met) and elucidate its regulatory role in neural progenitor cell (NPC) proliferation and differentiation, thereby uncovering the function of OTUD5 in neurodevelopment. Methods The OTUD5 variant was identified in two NDD patients via exome sequencing. Patient-derived induced pluripotent stem cells (iPSCs) and CRISPR/Cas9-corrected isogenic controls were generated. NPC proliferative activity was assessed by Ki67 immunofluorescence staining, cell-cycle distribution was analyzed by flow cytometry, and neuronal differentiation was evaluated by Tuj1/MAP2 immunofluorescence staining. Substrate screening was conducted in HEK293T cells using co-immunoprecipitation (Co-IP) and mass spectrometry. Deubiquitination capacity and protein stability were validated through ubiquitination assays and cycloheximide (CHX) chase experiments. Results The p.Val233Met variant, located within the catalytic OTU domain, induced a marked conformational alteration in the OTUD5 protein. Functionally, the variant caused aberrant NPC proliferation (1.8-fold increase in Ki67+ cells, accompanied by release of G1 arrest) and impaired neuronal differentiation (60% reduction in Tuj+ cells). Mechanistically, wild-type OTUD5 stabilized GSK3β by removing K48-linked ubiquitin chains, whereas the mutant isoform exhibited diminished deubiquitinase activity, accelerating GSK3β degradation and shortening its half-life by 40%. Conclusion This study establishes a novel disease mechanism whereby OTUD5 mutations disrupt NPC homeostasis through GSK3β destabilization, highlighting the critical role of ubiquitination regulation in neurodevelopment. Our iPSC model provides a platform for testing GSK3β-targeted therapies in OTUD5-related NDDs.
Stem Cell Research & Therapy•2025•DOI: 10.1186/s13287-025-04463-7
Background: Recent advances in clinical trials have involved the transplantation of induced retinal pigment epithelium (iRPE) cells from stem cells in creating a functional monolayer that mimics the characteristics of natural adult RPE cells. One method of achieving this goal is through the use of tissue engineering. In this research, decellularised femtosecond laser intrastromal lenticules (dfLEN) were employed as a scaffold for cultivating a bioengineered iRPE monolayer sheet. Methods: iRPE cells were obtained by differentiating induced pluripotent stem cells (iPSC). These cells were then seeded on decellularized FLI-lenticules (dfLEN). The functionality, characterization, and oxidative stress of iRPE cultured on dfLEN were compared with those cultured on plates (TCP) using various assays such as immunofluorescence (IF), Edu, CCK8, ELISA, DFCH-DA, and JC-1. Additionally, RNA-seq assays and electron microscope (SEM and TEM) were used to test the iRPE characteristic on engineered dfLEN. Finally, we evaluated the biocompatibility of iRPE-dfLEN sheets by transplanting them into the subretinal space of New Zealand white rabbits. Results: The iRPE cells cultured on dfLEN exhibited morphology and physiology similar to that of native RPE tissue. The dfLEN not only increased the resistance capacity of iRPE cells but also improved their functional properties compared to TCP. In addition, our results indicate that dfLEN enhances the expression of genes associated with cilium assembly, resulting in notable improvements in ciliogenesis in iRPE cells. Finally, the dfLEN-iRPE sheets demonstrated favorable biocompatibility and some viability when transplanted into the subretinal space of rabbits for a period of 14 days.
Stem Cell Research & Therapy•2025•DOI: 10.1186/s13287-025-04480-6
Background Diabetic foot ulcers (DFU) typically exhibit impaired healing due to dysregulated re-epithelialization and excessive inflammation. Succinate, a key metabolic intermediate, is now understood to regulate inflammation through G Protein-Coupled Receptor 91 (GPR91) and succinate dehydrogenase (SDH), although its role in DFU remains unclear. Methods Co-cultures of M2 macrophages and epithelial cells, along with clinical samples, were used to analyze the expression of GPR91 and SDH. Functional assays were performed using high glucose (HG)-treated M2 macrophages (HG-M2) and an in vivo model. Cytokine and growth factor levels in cell supernatant were measured, and molecular mechanisms were explored via qRT-PCR, flow cytometry, and western blot analysis. Results Elevated glucose concentrations increased succinate levels and disrupted M2 macrophage–epidermal stem cells (EpSCs) interactions. GPR91 knockdown worsened HG-M2 dysfunction, while GPR91 overexpression (OE-GPR91) enhanced anti-inflammatory responses and reduced succinate. OE-GPR91-conditioned medium preserved EpSCs stemness and promoted migration mediated by hepatocyte growth factor (HGF). SDH inhibition (via Dimethyl malonate, DMM) boosted M2 macrophage activity by reducing reactive oxygen species (ROS) and upregulating Gpr91 expression. Mechanistically, GPR91 activated the pAkt/pGSK3β/β-catenin pathway, while DMM enhanced M2 macrophage function via the PI3K-Akt/pERK1/2 pathway. Conclusions GPR91 upregulation and SDH inhibition improve HG-M2 macrophage function, reduce inflammation, and enhance HGF-mediated EpSCs repair. Targeting both pathways may represent a promising approach to promote DFU healing.
Stem Cell Research & Therapy•2025•DOI: 10.1186/s13287-025-04764-x
Background Human hair follicle dermal sheath cup cells (DSCCs) hold promise as a cell source of regenerative medicine treatment for hair loss owing to their ability to secrete growth factors and/or signal pathway activators. The therapeutic effect of autologous DSCCs transplantation for male/female pattern hair loss (PHL) was demonstrated in a phase III equivalent clinical study. Intralesional inflammation has been implicated in the pathophysiology of various hair loss diseases, including PHL. As DSCCs possess mesenchymal stem/stromal cell (MSC)-like properties and MSCs are immunosuppressive, we investigated whether they exhibit immunoregulatory capabilities comparable to MSCs and developed an in vitro morphometric assay to predict this capability. Methods DSCCs were isolated via microdissection and propagated in vitro. Their conformity to MSC criteria was assessed based on cell surface antigen expression and differentiation potential. Furthermore, immunoregulatory capabilities were assessed by co-culturing DSCCs with anti-CD3/28 antibody-stimulated peripheral blood mononuclear cells (PBMCs) and examining the suppression mechanisms through pharmacological intervention. Multiple lots of DSCCs derived from various donors and manufacturing conditions were cultured and analyzed by phase-contrast microscopy to obtain their morphometric profiles. Parameters correlating with the expression levels of immunomodulatory factors were used to create a predictive model. Additional DSCC lots were manufactured to validate the predictive model. Results Similar to MSCs, cell differentiation assays revealed that DSCCs exhibited multipotency, and they did not express co-stimulatory molecules in response to immunogenic stimuli, suggesting low immunogenicity. Moreover, co-culture experiments with allogeneic PBMCs revealed that DSCCs reduced T cell proliferation (from 78 to 5%) and
Stem Cell Research & Therapy•2024•DOI: 10.1186/s13287-024-03838-6
Correction to: Stem Cell Research & Therapy (2023) 14:17. The authors note that during the preparation of the manuscript, the track plot included the expression trends of 14 genes (with Tk1 and Pclaf repeated twice), but only 13 gene symbols were labeled, resulting in a mismatch and repeat of the trackplot with the image on the right. This error occurred during the typesetting process of the original figures. The authors have corrected the annotations in Fig. 2C as shown ahead in this correction article, apologise for the error, and confirm that the overall results and conclusions are not affected by this change.
Stem Cell Research & Therapy•2024•DOI: 10.1186/s13287-024-03799-w
Human hematopoietic stem cell (HSC)-transferred humanized mice are valuable models for exploring human hematology and immunology. However, sufficient recapitulation of human hematopoiesis in mice requires large quantities of enriched human CD34+ HSCs and total-body irradiation for adequate engraftment. Recently, we generated a NOG mouse strain with a point mutation in the c-kit tyrosine kinase domain (W41 mutant; NOGW mice). In this study, we examined the ability of NOGW mice to reconstitute human hematopoietic cells. Irradiated NOGW mice exhibited high engraftment levels of human CD45+ cells in the peripheral blood, even when only 5,000–10,000 CD34+ HSCs were transferred. Efficient engraftment of human CD45+ cells was also observed in non-irradiated NOGW mice transferred with 20,000–40,000 HSCs. The bone marrow (BM) of NOGW mice exhibited significantly more engrafted human HSCs or progenitor cells (CD34+CD38− or CD34+CD38+ cells) than the BM of NOG mice. Furthermore, we generated a human cytokine (interleukin-3 and granulocyte-macrophage colony-stimulating factor) transgenic NOG-W41 (NOGW-EXL) mouse to achieve multilineage reconstitution with sufficient engraftment of human hematopoietic cells. Non-irradiated NOGW-EXL mice showed significantly higher engraftment levels of human CD45+ and myeloid lineage cells, particularly granulocytes and platelets/megakaryocytes, than non-irradiated NOGW or irradiated NOG-EXL mice after human CD34+ cell transplantation. Serial BM transplantation experiments revealed that NOGW mice exhibited the highest potential for long-term HSC compared with other strains. Consequently, c-kit mutant NOGW-EXL humanized mice represent an advanced model for HSC-transferred humanized mice and hold promise for widespread applications owing to their high versatility.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2026030
The recombination-activating gene (RAG)-mediated V(D)J rearrangement mechanism is essential for generating the diversity of the adaptive immune system in jawed vertebrates. The RAG1 and RAG2 proteins form a tetrameric complex that recognizes and cleaves pairs of recombination signal sequences (RSSs). In jawed vertebrates, the N-terminal region of RAG1 and the C-terminal region of RAG2 are critical for the regulation of physiological function and enhancement of recombination accuracy. Moreover, the intracellular activity of RAG1/2 is rigorously regulated and is thought to represent the aspect shaped by host coevolution. However, these regulatory mechanisms remain incompletely understood, impeding our deeper understanding of the physiological functions of RAG. Previous studies have shown that jawed vertebrate RAG evolved from invertebrate RAG-like (RAGL) transposases and underwent key structural changes during evolution. RAGL genes have been identified in multiple invertebrate species, but from Branchiostoma belcheri (Bb), the RAG-like protein in Bb amphioxus (BbRAG1L) and the RAG2-like protein in Bb amphioxus (BbRAG2L) are among the most extensively characterized to date. Lancelets (amphioxus) represent the most basal extant chordates (cephalochordates) that diverged from the other two chordate lineages (urochordates and vertebrates). Consequently, amphioxus has been considered a “living fossil” for studying the origins and evolution of adaptive immunity. Homologs of the core structural domain of RAG1 were discovered in the amphioxus genome in 2005, and the homologs were subsequently shown to exhibit recombinase activity in cooperation with murine-derived RAG2. A large number of transposon genes were then identified in the amphioxus genome, followed by the identification of target site duplication (TSD) and terminal inverted repeat (TIR) sequences in the amphioxus genome. Notably, BbRAG2L lacks the C-terminal region present in murine RAG2, including the acidic hinge and PHD domain. BbRAG1L coupled with BbRAG2L displays significant transposition activity in vitro and in human cell lines. In contrast, vertebrate RAG exhibits limited transposon activity in vitro and extremely low transposon activity in vivo. This leads to the question of how RAG lost its transposase activity during evolution. Structural comparison and functional experiments of BbRAGL (protoRAG) and vertebrate RAG have revealed that vertebrates undergo essential adaptive changes to eliminate the primitive transposase activity of protoRAG. Research on BbRAGL has therefore provided strong support for the hypothesis that RAG originated from a domesticated transposon. Given the similarity in both structure and function between BbRAG1L and vertebrate RAG and the fact that BbRAGL proteins are the only RAGL proteins that can currently be efficiently expressed in vertebrate cells, characterizing the binding partners of BbRAGL involved in amphioxus is of great significance for understanding the regulation of BbRAGL and its adaptation and evolution. Parallel comparison of BbRAGL and RAG interaction networks may further clarify how BbRAGL transitioned from a transposase to a recombinase during host coevolution and reveal general principles of vertebrate intracellular RAG regulation. In recent years, proximity labeling techniques have greatly facilitated approaches to identifying RAG cofactors. For example, RAG1 fused to biotin ligase RAG1 fused to a biotin ligase found in Escherichia coli (BirM) has been used to define dynamic interaction networks of RAG-associated proteins. Inspired by these advances, we sought to apply a similar approach to identify the BbRAGL cofactors in amphioxus and build a regulatory network of BbRAGL that could shed light on its relationship with host evolution. However, endogenous BbRAGL expression is undetectable in the adult amphioxus (data not shown), and the generation of transgenic amphioxus lines expressing proximity-based labeling proteins remains technically challenging. These constraints make it difficult to analyze native cofactors of endogenous BbRAGL in vivo. Because BbRAGL can be expressed in vertebrate cell lines, we instead developed an in vitro proximity-labeling strategy. We fused TurboID (TbID), an engineered biotin ligase, to the N-terminus of BbRAG1L or BbRAG2L (Figure 1A), purified the fusion proteins, and incubated them with protein lysates prepared from the hepatic cecum and colons of adult B. belcheri amphioxus in vitro to label the interacting proteins. The proteins were finally analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) (Figure 1B). A pTT5 vector was used to express plasmids encoding maltose-binding protein (MBP) and TurboID with or without full-length BbRAG1L or BbRAG2L, yielding pTT5-MBP-TurboID, pTT5-MBP-TurboID-BbRAG1L, and pTT5-MBP-TurboID-BbRAG2L. Plasmids expressing MBP-BbRAG1L and MBP-BbRAG2L were also generated.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2025206
Anti-aging foods not only benefit elderly individuals but also drive the development of safe and effective natural drugs. Here, we report that the addition of total flavonoids of litchi seed (TFL) delays replicative senescence and stress-induced senescence. TFL alleviates the senescence-associated secretory phenotype (SASP) and reduces the degree of DNA damage caused by bleomycin (BLM). TFL also counteracts stress-induced pulmonary senescence and fibrosis. TFL reduces the protein level of p21 in mouse lung and alleviates pulmonary fibrosis. Transcriptome profiling further reveals that TFL plays a key role in its anti-aging mechanism by inhibiting the SASP. Mechanistically, TFL suppresses p65 protein expression, thereby inhibiting IL-1α and IL-1β and delaying cellular senescence. Gut microbiome analysis reveals that the abundance and functions of the mouse gut microbiome change after BLM exposure and that TFL treatment reverses these changes. Overall, we provide a theoretical basis for the future application of TFL as a potential anti-aging product.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2026018
Doxorubicin (DOX) remains a cornerstone chemotherapeutic agent for malignancies, yet its clinical utility is severely limited by dose-dependent cardiotoxicity, which can lead to progressive left ventricular dysfunction and heart failure. Pleiotrophin (PTN), a heparin-binding growth factor with diverse physiological functions, regulates glucose and lipid metabolism and promotes oxidative energy pathways. However, whether PTN exerts protective effects against DOX-induced cardiotoxicity (DIC) remains unclear. In this study, we establish cellular and animal models of DIC. DOX administration induces pronounced myocardial injury in both models, characterized by impaired ventricular contractility, increased fibrotic remodeling, and reduced cell viability. Concurrently, PTN protein expression is significantly downregulated in cardiomyocytes under DOX treatment. Overexpression of PTN substantially alleviates these pathological changes. In vitro, PTN reduces mitochondrial oxidative stress and apoptosis while restoring energy production and cell viability. In vivo, PTN improves mitochondrial ultrastructure, decreases cardiomyocyte apoptosis, and enhances cardiac function. Mechanistically, PTN directly binds to SIRT1 and activates AMPK phosphorylation at Thr172, triggering a downstream cascade through the AMPK-PGC1α axis that reprograms mitochondrial energy metabolism and attenuates cardiotoxicity. In conclusion, the PTN-SIRT1 axis protects against DIC by reducing oxidative stress and promoting mitochondrial energy homeostasis via the AMPK/PGC1α pathway, highlighting its potential as a novel therapeutic target for preventing chemotherapy-related cardiac injury.
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•2024•DOI: 10.3724/abbs.2024134
SUN5, a testis-specific gene, is associated with acephalic spermatozoa syndrome (ASS). Here, we demonstrate that SUN5 is involved in mRNA export. In Sun5-knockout mice (Sun5–/–), poly(A)+ RNA accumulates in the nuclei of germ cells, leading to reduced sperm counts, decreased sperm motility and disrupted sperm head-to-tail junctions. Additionally, in the GC-2 germ cell line with RNA interference of Sun5, heterogeneous nuclear ribonucleoproteins (hnRNPs) and poly (A)+ RNA (mainly mRNA) are retained in the nucleus. Further mechanistic studies reveal that SUN5 interacts with Nxf1 (nuclear RNA export factor 1) and nucleoporin 93 (Nup93). Interference with Nup93 inhibits mRNA export. Treatment with leptomycin B to block the CRM1 pathway indicates that Sun5 regulates mRNA export through an Nxf1-dependent pathway. In Sun5–/– mice, the binding of Nxf1 and Nup93 decreases due to loss of Sun5 function, and the process of submitting Nxf1-binding mRNPs to Nup93 is inhibited, resulting in abnormal spermatogenesis. Together, these data may elucidate a novel pathway for mRNA export in male germ cells.
Acta Biochimica et Biophysica Sinica•2024•DOI: 10.3724/abbs.2024078
Neural tube defects (NTDs) are characterized by the failure of neural tube closure during embryogenesis and are considered the most common and severe central nervous system anomalies during early development. Recent microRNA (miRNA) expression profiling studies have revealed that the dysregulation of several miRNAs plays an important role in retinoic acid (RA)-induced NTDs. However, the molecular functions of these miRNAs in NTDs remain largely unidentified. Here, we show that miR-10a-5p is significantly upregulated in RA-induced NTDs and results in reduced cell growth due to cell cycle arrest and dysregulation of cell differentiation. Moreover, the cell adhesion molecule L1-like (Chl1) is identified as a direct target of miR-10a-5p in neural stem cells (NSCs) in vitro, and its expression is reduced in RA-induced NTDs. siRNA-mediated knockdown of intracellular Chl1 affects cell proliferation and differentiation similar to those of miR-10a-5p overexpression, which further leads to the inhibition of the expressions of downstream ERK1/2 MAPK signaling pathway proteins. These cellular responses are abrogated by either increased expression of the direct target of miR-10a-5p (Chl1) or an ERK agonist such as honokiol. Overall, our study demonstrates that miR-10a-5p plays a major role in the process of NSC growth and differentiation by directly targeting Chl1, which in turn induces the downregulation of the ERK1/2 cascade, suggesting that miR-10a-5p and Chl1 are critical for NTD formation in the development of embryos.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2025213
Myocardial infarction (MI) causes irreversible cardiomyocyte loss, creating a need for cardiac repair therapies. The role of cell division cycle 5-like (CDC5L), a cell cycle regulator, in cardiac repair is unknown. This study aims to define the role of CDC5L in mitigating ischemia-reperfusion (I/R) injury by assessing its impact on cardiomyocyte proliferation and apoptosis and to determine the mechanism involving the FGF10-YAP axis. We model cardiac injury using in vitro oxygen-glucose deprivation/reoxygenation (OGD/R) in neonatal mouse cardiomyocytes and in vivo I/R in adult mice. To investigate CDC5L function, we modulate its expression via adenoviral or AAV9-mediated overexpression or knockdown. Proliferation markers (EdU+, Ki67+, pH3+), apoptosis (TUNEL staining, Bax/Bcl-2 ratio), and cardiac function (echocardiography) are assessed. Through transcriptomic screening, we identify CDC5L downstream targets and validate their functional roles using FGF10 knockdown rescue assays. We find that CDC5L is upregulated in the post-I/R murine myocardium. Its overexpression enhances cardiomyocyte proliferation, preserves cardiac function, reduces apoptosis, and diminishes infarct size. Transcriptomic analysis identifies FGF10 as a key downstream effector, and we confirm that CDC5L upregulates FGF10 expression. Notably, FGF10 knockdown reverses the proliferative and anti-apoptotic effects of CDC5L. Moreover, the CDC5L-mediated reduction in YAP phosphorylation is also dependent on FGF10, as this effect is abolished upon FGF10 knockdown. In conclusion, CDC5L attenuates cardiac I/R injury by promoting cardiomyocyte proliferation and inhibiting apoptosis through the FGF10-YAP pathway. This CDC5L-FGF10-YAP axis represents a promising therapeutic target to improve myocardial regeneration and recovery after myocardial infarction.
Acta Biochimica et Biophysica Sinica•2024•DOI: 10.3724/abbs.2024056
Biological control of pests and pathogens has attracted much attention due to its green, safe and effective characteristics. However, it faces the dilemma of insignificant effects in large-scale applications. Therefore, an in-depth exploration of the metabolic potential of biocontrol fungi based on big omics data is crucial for a comprehensive and systematic understanding of the specific modes of action operated by various biocontrol fungi. This article analyzes the preferences for extracellular carbon and nitrogen source degradation, secondary metabolites (nonribosomal peptides, polyketide synthases) and their product characteristics and the conversion relationship between extracellular primary metabolism and intracellular secondary metabolism for eight different filamentous fungi with characteristics appropriate for the biological control of bacterial pathogens and phytopathogenic nematodes. Further clarification is provided that Paecilomyces lilacinus, encoding a large number of hydrolase enzymes capable of degrading pathogen protection barrier, can be directly applied in the field as a predatory biocontrol fungus, whereas Trichoderma, as an antibiosis-active biocontrol control fungus, can form dominant strains on preferred substrates and produce a large number of secondary metabolites to achieve antibacterial effects. By clarifying the levels of biological control achievable by different biocontrol fungi, we provide a theoretical foundation for their application to cropping habitats.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2024120
In this study we investigate the role of Zipper-interacting protein kinase (ZIPK) in high glucose-induced vascular injury, focusing on its interaction with STAT5A and its effects on p53 and inducible nitric oxide synthase (NOS2) expression. Human umbilical vein endothelial cells (HUVECs) are cultured under normal (5 mM) and high (25 mM) glucose conditions. Protein and gene expression levels are assessed by western blot analysis and qPCR respectively, while ROS levels are measured via flow cytometry. ZIPK expression is manipulated using overexpression plasmids, siRNAs, and shRNAs. The effects of the ZIPK inhibitor TC-DAPK6 are evaluated in a diabetic rat model. Our results show that high glucose significantly upregulates ZIPK, STAT5A, p53, and NOS2 expressions in HUVECs, thus increasing oxidative stress. Silencing of STAT5A reduces p53 and NOS2 expressions and reactive oxygen species (ROS) accumulation. ZIPK is essential for high glucose-induced p53 expression and ROS accumulation, while silencing of ZIPK reverses these effects. Overexpression of ZIPK combined with STAT5A silencing attenuates glucose-induced alterations in p53 and NOS2 expression, thereby preventing cell damage. Coimmunoprecipitation reveals a direct interaction between ZIPK and STAT5A in the nucleus under high-glucose condition. In diabetic rats, TC-DAPK6 treatment significantly decreases ZIPK, p53, and NOS2 expressions. Our findings suggest that ZIPK plays a critical role in high glucose-induced vascular injury via STAT5A-mediated pathways, proposing that ZIPK is a potential therapeutic target for diabetic vascular complications.
Acta Biochimica et Biophysica Sinica•2024•DOI: 10.3724/abbs.2024110
Immune checkpoint inhibitors (ICIs) targeting programmed cell death 1/programmed cell death ligand-1 (PD-1/PD-L1) have significantly prolonged the survival of advanced/metastatic patients with lung cancer. However, only a small proportion of patients can benefit from ICIs, and clinical management of the treatment process remains challenging. Glycosylation has added a new dimension to advance our understanding of tumor immunity and immunotherapy. To systematically characterize anti-PD-1/PD-L1 immunotherapy-related changes in serum glycoproteins, a series of serum samples from 12 patients with metastatic lung squamous cell carcinoma (SCC) and lung adenocarcinoma (ADC), collected before and during ICIs treatment, are firstly analyzed with mass-spectrometry-based label-free quantification method. Second, a stratification analysis is performed among anti-PD-1/PD-L1 responders and non-responders, with serum levels of glycopeptides correlated with treatment response. In addition, in an independent validation cohort, a large-scale site-specific profiling strategy based on chemical labeling is employed to confirm the unusual characteristics of IgG N-glycosylation associated with anti-PD-1/PD-L1 treatment. Unbiased label-free quantitative glycoproteomics reveals serum levels’ alterations related to anti-PD-1/PD-L1 treatment in 27 out of 337 quantified glycopeptides. The intact glycopeptide EEQFN177STYR (H3N4) corresponding to IgG4 is significantly increased during anti-PD-1/PD-L1 treatment (FC=2.65, P=0.0083) and has the highest increase in anti-PD-1/PD-L1 responders (FC=5.84, P=0.0190). Quantitative glycoproteomics based on protein purification and chemical labeling confirms this observation. Furthermore, obvious associations between the two intact glycopeptides (EEQFN177STYR (H3N4) of IgG4, EEQYN227STFR (H3N4F1) of IgG3) and response to treatment are observed, which may play a guiding role in cancer immunotherapy. Our findings could benefit future clinical disease management.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2024163
Human rhomboid family-1 (RHBDF1) gene is recognized as an oncogene involved in breast cancer development. Previous studies have indicated that RHBDF1 contributes significantly to endoplasmic reticulum (ER) protein homeostasis by stabilizing the binding immunoglobulin protein (BiP) and promoting the unfolded protein response (UPR). Here, we report a relationship between RHBDF1 and the ER stress sensors PERK, IRE1, and ATF6. We show that RHBDF1 deficiency in breast cancer cells results in decreased levels of PERK, pPERK, and peIF2α. These protein levels can be restored in RHBDF1-deficient breast cancer cells by artificial overexpression of RHBDF1 but not IRE1 or ATF6. Additionally, we show that the transcription factor FoxO3 is essential for the RHBDF1-mediated production of PERK. Subsequent analysis reveals that RHBDF1 activates JNK, which causes FoxO3 to translocate into the cell nucleus. These findings demonstrate that RHBDF1 supports the UPR by upregulating the PERK/peIF2α pathway via the JNK/FoxO3 axis and that the functions of RHBDF1 are essential for preserving the homeostasis of ER proteins.
Stem Cell Research & Therapy•2026•DOI: 10.1186/s13287-026-04995-6
Background Gastrointestinal diseases often involve cellular damage, degeneration or dysfunction in the tract, frequently requiring surgical interventions risking complications and lowered quality of life. Regenerative medicine holds great promise in improving patient care and providing novel treatment options for previously irreparable and untreatable tissues. Despite the clinical potential of intestinal organoids as a resource for regenerative cell therapy and bioengineering, the lack of clinical-grade cultures has hampered further development. Moreover, strategies to efficiently and reliably expand clinical-grade cultures at the scale required for application is limited. Methods A GMP-compliant protocol was developed to generate patient-derived colonic organoids from endoscopic biopsies. Clinical-grade colonic organoids cultured and expanded in Type-I collagen were compared to conventional Matrigel cultured organoids. To improve the culture-, cost-, and time-efficiency of culture expansion, several strategies were developed including organoid area-based passaging, one well plate culture, and the incorporation of Wnt activating peptide, PG-008. Conventional recombinant WNT3A culture was compared to the peptide PG-008 culture using single cell RNA sequencing. Results Clinical-grade collagen cultured organoids exhibited similar culture efficiency to Matrigel. Organoid establishment rate from 60 patients using the GMP-compliant protocol was 82%. The incorporation of PG-008 significantly enhanced organoid growth and stabilized patient-patient variability through intestinal stem cell (ISC) enrichment. Single cell RNA sequencing revealed that PG-008 resulted in remarkably pure culture consisting of ISCs. Peptide-based Wnt signal activation enables scalable production of clinical-grade patient-derived intestinal organoids for regenerative cell therapy. Intriguingly, our GMP-grade colonic organoids contained LGR5+ ISCs, and injury-induced LGR5− regenerative ISCs, both enriched in peptide culture. Conclusions Our study establishes clinical-grade colonic organoids for further application, including autologous transplantations and bioengineering. Further, collagen cultured organoids can be a valuable model facilitating in vitro investigation.
Stem Cell Research & Therapy•2026•DOI: 10.1186/s13287-026-04974-x
Background: Variants in OTUD5 are associated with neurodevelopmental disorders (NDDs), yet the underlying molecular mechanisms remain unclear. This study aimed to investigate the pathogenicity of a novel OTUD5 variant (c.697G>A, p.Val233Met) and elucidate its regulatory role in neural progenitor cell (NPC) proliferation and differentiation, thereby uncovering the function of OTUD5 in neurodevelopment. Methods: The OTUD5 variant was identified in two NDD patients via exome sequencing. Patient-derived induced pluripotent stem cells (iPSCs) and CRISPR/Cas9-corrected isogenic controls were generated. NPC proliferative activity was assessed by Ki67 immunofluorescence staining, cell-cycle distribution was analyzed by flow cytometry, and neuronal differentiation was evaluated by Tuj1/MAP2 immunofluorescence staining. Substrate screening was conducted in HEK293T cells using co-immunoprecipitation (Co-IP) and mass spectrometry. Deubiquitination capacity and protein stability were validated through ubiquitination assays and cycloheximide (CHX) chase experiments. Results: The p.Val233Met variant, located within the catalytic OTU domain, induced a marked conformational alteration in the OTUD5 protein. Functionally, the variant caused aberrant NPC proliferation (1.8-fold increase in Ki67+ cells, accompanied by release of G1 arrest) and impaired neuronal differentiation (60% reduction in Tuj+ cells). Mechanistically, wild-type OTUD5 stabilized GSK3β by removing K48-linked ubiquitin chains, whereas the mutant isoform exhibited diminished deubiquitinase activity, accelerating GSK3β degradation and shortening its half-life by 40%. Conclusion: This study establishes a novel disease mechanism whereby OTUD5 mutations disrupt NPC homeostasis through GSK3β destabilization, highlighting the critical role of ubiquitination regulation in neurodevelopment. Our iPSC model provides a platform for testing GSK3β-targeted therapies in OTUD5-related NDDs.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2026018
Doxorubicin (DOX) remains a cornerstone chemotherapeutic agent for malignancies, yet its clinical utility is severely limited by dose-dependent cardiotoxicity, which can lead to progressive left ventricular dysfunction and heart failure. Pleiotrophin (PTN), a heparin-binding growth factor with diverse physiological functions, regulates glucose and lipid metabolism and promotes oxidative energy pathways. However, whether PTN exerts protective effects against DOX-induced cardiotoxicity (DIC) remains unclear. In this study, we establish cellular and animal models of DIC. DOX administration induces pronounced myocardial injury in both models, characterized by impaired ventricular contractility, increased fibrotic remodeling, and reduced cell viability. Concurrently, PTN protein expression is significantly downregulated in cardiomyocytes under DOX treatment. Overexpression of PTN substantially alleviates these pathological changes. In vitro, PTN reduces mitochondrial oxidative stress and apoptosis while restoring energy production and cell viability. In vivo, PTN improves mitochondrial ultrastructure, decreases cardiomyocyte apoptosis, and enhances cardiac function. Mechanistically, PTN directly binds to SIRT1 and activates AMPK phosphorylation at Thr172, triggering a downstream cascade through the AMPK-PGC1α axis that reprograms mitochondrial energy metabolism and attenuates cardiotoxicity. In conclusion, the PTN-SIRT1 axis protects against DIC by reducing oxidative stress and promoting mitochondrial energy homeostasis via the AMPK/PGC1α pathway, highlighting its potential as a novel therapeutic target for preventing chemotherapy-related cardiac injury.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2025206
Anti-aging foods not only benefit elderly individuals but also drive the development of safe and effective natural drugs. Here, we report that the addition of total flavonoids of litchi seed (TFL) delays replicative senescence and stress-induced senescence. TFL alleviates the senescence-associated secretory phenotype (SASP) and reduces the degree of DNA damage caused by bleomycin (BLM). TFL also counteracts stress-induced pulmonary senescence and fibrosis. TFL reduces the protein level of p21 in mouse lung and alleviates pulmonary fibrosis. Transcriptome profiling further reveals that TFL plays a key role in its anti-aging mechanism by inhibiting the SASP. Mechanistically, TFL suppresses p65 protein expression, thereby inhibiting IL-1α and IL-1β and delaying cellular senescence. Gut microbiome analysis reveals that the abundance and functions of the mouse gut microbiome change after BLM exposure and that TFL treatment reverses these changes. Overall, we provide a theoretical basis for the future application of TFL as a potential anti-aging product.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2026030
The recombination-activating gene (RAG)-mediated V(D)J rearrangement is essential for adaptive immunity in jawed vertebrates. RAG evolved from an invertebrate RAG-like (RAGL) transposase, with the amphioxus protoRAG (BbRAG1L/BbRAG2L) serving as a key model. However, the regulatory mechanisms of protoRAG remain unclear. Here, we developed an in vitro proximity labeling assay using TurboID fused to BbRAG1L or BbRAG2L to identify interacting proteins from amphioxus hepatic cecum and colon lysates. The fusion proteins were expressed in Expi293F cells, purified, and incubated with amphioxus lysates in the presence of biotin and ATP. Biotinylated proteins were enriched via streptavidin beads and analyzed by LC-MS/MS. This approach enables the identification of protoRAG-interacting proteins without the need for transgenic amphioxus, providing a valuable tool to study the evolution of RAG regulation.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21205
BACKGROUND: Macrophage polarization demonstrates significant potential in disease treatment, particularly in areas such as cancer, inflammation, and autoimmune diseases. Establishing standardized in vitro models can lay the groundwork for in-depth research into the mechanisms of macrophage polarization. OBJECTIVE: To observe the in vitro growth characteristics of bone marrow-derived macrophages from C57BL/6 mice and to establish a standardized in vitro model for M1 and M2 macrophage polarization. METHODS: Femurs and tibias of C57BL/6 mice were aseptically separated, and the contents of the bone marrow cavity were collected. After filtering through a mesh and lysing erythrocytes, the contents were resuspended in high-glucose DMEM containing 20 ng/mL macrophage colony-stimulating factor and inoculated in 6-well plates according to experimental requirements. On day 7, they were differentiated into mature mouse bone marrow-derived macrophages (M0 type). Then, 100 ng/mL lipopolysaccharide was used to induce polarization to M1 type, and 20 ng/mL interleukin-4 was used to induce polarization to M2 type. Flow cytometry and RT-qPCR were used to detect the expression of corresponding markers in macrophages under different polarization states, and Western blot was used to detect the expression of M1 macrophage marker pathway proteins p-STAT1, STAT1 and M2 macrophage marker pathway proteins p-STAT6, STAT6. RESULTS AND CONCLUSION: (1) After stimulation with 20 ng/mL macrophage colony-stimulating factor for 7 days, flow cytometry showed that the positive rate of macrophage surface marker F4/80 reached 98.1%. (2) After stimulation with 100 ng/mL lipopolysaccharide for 6 h, the positive rates of F4/80 and CD86 were about 35%, and RT-qPCR showed that the mRNA expression of M1 macrophage markers inducible nitric oxide synthase, interleukin-6, macrophage inflammatory protein 1α, and monocyte chemoattractant protein 1 were significantly higher than those in the control group (P < 0.01). (3) After stimulation with 20 ng/mL interleukin-4 for 24 h, the mean fluorescence intensity of CD206 was significantly increased, and RT-qPCR showed that the mRNA expression of M2 macrophage markers Chi3l3 (Ym1), interleukin-10, and arginase 1 were significantly higher than those in the control group (P < 0.01). (4) Western blot results showed that lipopolysaccharide-induced M1 macrophage marker pathway protein p-STAT1 was significantly activated; interleukin-4-induced M2 macrophage marker pathway protein p-STAT6 was significantly activated. These results indicate that lipopolysaccharide and interleukin-4 effectively induced polarization of bone marrow-derived macrophages to M1 and M2 types, respectively.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21239
BACKGROUND: Estrogen deficiency can lead to a decrease in skeletal muscle mass and muscle strength in postmenopausal women, thereby affecting their quality of life. Muscle mass is maintained by satellite cells, which are regulated by estrogen. Regular exercise, especially high impact exercise (such as resistance training and high-intensity interval training), can induce muscle hypertrophy, but the role and mechanism of estrogen are still unclear. OBJECTIVE: To explore the effects of high-intensity interval training combined with estrogen therapy on skeletal muscle hypertrophy in ovariectomized rats and reveal its possible mechanism. METHODS: Sixty 8-week-old female Sprague-Dawley rats were divided into five groups using a random number table method: sham operation, model sedentary group, model exercise group, model hormone group, or model combined group. Bilateral ovariectomy was used to establish an estrogen deficiency model. Twelve weeks after operation, the model exercise and model combined group performed high-intensity interval training for 8 weeks (3 times/week), and hormone treatment groups received abdominal subcutaneous injection of 17β-estradiol (once a day for 8 weeks). Seventy-two hours after the last training, the grip force of the forelimb was measured by an electronic grip force meter. The gastrocnemius muscle was separated, and muscle mass index was calculated as muscle mass/body mass ratio. Hematoxylin-eosin staining was used to obtain cell cross-sectional area. Immunofluorescence staining was used to classify muscle fiber types and obtain myonuclear number, myonuclear domain size, and activated satellite cell number. BCA method was used to determine total protein concentration. Trizol method was used to extract total RNA. Western blot was used to detect ribosomal protein S6 expression. Real-time quantitative PCR was used to detect ribosomal RNA expression. RESULTS AND CONCLUSION: Compared with the sham operation group, the model sedentary group showed increased body mass and myosin heavy chain type I fiber proportion (P < 0.05), while uterine mass index, gastrocnemius mass index, grip strength, cell cross-sectional area, myosin heavy chain type IIa proportion, satellite cell and myonuclear number, myonuclear domain size, total protein and RNA content, and ribosomal protein S6, 18S rRNA and 28S rRNA expression decreased (P < 0.05). Compared with the model sedentary group, the model exercise group showed decreased body mass and myosin heavy chain type IIb proportion (P < 0.05), while uterine mass index, gastrocnemius mass index, grip strength, cell cross-sectional area, myosin heavy chain type IIa proportion, satellite cell number, myonuclear domain size, total protein and RNA content, and ribosomal protein S6 and 28S rRNA expression increased (P < 0.05). Compared with the model exercise group and model hormone group, the model combined group showed higher gastrocnemius mass index, grip strength, cell cross-sectional area, satellite cell number, myonuclear domain size, total protein and RNA content, and ribosomal protein S6 and 28S rRNA expression (P < 0.05). These results indicate that estrogen can enhance the skeletal muscle hypertrophy response induced by high-intensity interval training in ovariectomized rats, and the mechanism may be related to satellite cell activation, increased myonuclear domain and ribosome biogenesis, and improved ribosome function.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21303
OBJECTIVE: Current evidence indicates that exercise-induced oxidative stress involves a dual role of reactive oxygen species, which participate in exercise adaptation while potentially causing tissue damage, highlighting the necessity for precise regulation of antioxidant dosage and timing. This study employs a Meta-analytic approach to systematically evaluate the effects of antioxidant pretreatment on biomarkers of skeletal muscle oxidative stress injury following acute strenuous exercise, and to explore the moderating effects of dosage, intervention duration, and training status. METHODS: A systematic search was conducted for randomized controlled trials that investigated the effects of antioxidant pretreatment on exercise-induced oxidative stress in PubMed, Web of Science, EBSCO, CNKI, VIP and WanFang databases from inception to February 2025. Literature quality was assessed using the physiotherapy evidence database scale. Data analysis was performed using RevMan 5.4 and Stata statistical software. RESULTS: (1) This meta-analysis included 16 studies from 12 publications, comprising 264 athletes and regularly exercising individuals. (2) The physiotherapy evidence database scale scores ranged from 6-8 (7 studies) to 9 (5 studies), indicating overall high methodological quality. (3) Meta analysis results showed that antioxidant pretreatment significantly decreased post-exercise serum creatine kinase [standardized mean difference (SMD)=-0.31, 95% confidence interval (CI) (-0.63, 0.00), P=0.05], interleukin-6 [SMD=-0.66, 95%CI (-1.03, -0.29), P=0.0005], and malondialdehyde levels [SMD=-1.10, 95%CI (-1.96, -0.23), P=0.01], and increased glutathione peroxidase activity [SMD=1.33, 95%CI (0.87, 1.78), P < 0.00001] and total antioxidant capacity [MD=4.77, 95%CI (3.87, 5.67), P < 0.00001]. Subgroup analysis showed that low-dose (≤500 mg/d) short-term (≤14 d) intervention had a more significant inhibitory effect on malondialdehyde levels (SMD=-1.15), while high-dose long-term intervention may inhibit exercise adaptation. Training status significantly moderated the effect size, with general athletes showing greater reduction in malondialdehyde levels than elite athletes (P < 0.05). CONCLUSION: Antioxidant pretreatment can effectively alleviate oxidative stress damage induced by acute strenuous exercise, but its effect is influenced by dosage, intervention duration, and training status. Short-term high-dose supplementation is suitable for rapid recovery during competition periods, while long-term application requires weighing antioxidant benefits against the risk of adaptation inhibition.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21287
BACKGROUND: The onset of cervical instability in middle-aged and young adults often begins with neck muscle injuries. A deeper understanding of changes in neck muscles during cervical instability and their correlation can provide valuable data to support the prevention and treatment of cervical instability in this population. OBJECTIVE: To explore the correlation between cervical instability and neck muscle changes in middle-aged and young adults. METHODS: A total of 98 patients with cervical C4/5 instability and 88 healthy subjects, aged 18-45 years, were enrolled through recruitment advertisements and the Department of Spine, Wangjing Hospital, China Academy of Traditional Chinese Medicine. Cervical X-rays were collected to measure cervical curvature and C4/5 vertebral angular displacement. Cervical magnetic resonance imaging was taken to obtain data on C4/5 intervertebral disc signal intensity, as well as the relative cross-sectional area and fat ratio of neck muscles, including prevertebral muscles, deep posterior cervical muscles, and superficial muscles. A univariate intergroup comparison of X-ray and magnetic resonance imaging data was conducted between cervical instability subjects and healthy controls, along with Spearman correlation analysis between C4/5 angular displacement and disc signal intensity, relative cross-sectional area of neck muscles and fat percentage at the C4/5 level in cervical instability patients. RESULTS AND CONCLUSION: The cervical instability group had significantly greater age, C4/5 horizontal displacement, C4/5 angular displacement, and fat ratio of deep posterior cervical muscles than the healthy group (P < 0.05), while cervical curvature and relative cross-sectional area of deep posterior cervical muscles were significantly smaller (P < 0.05). Spearman correlation analysis showed a negative correlation between C4/5 angular displacement and relative cross-sectional area of deep posterior cervical muscles (P < 0.05). These findings suggest that changes in deep posterior cervical muscles may be closely related to the occurrence of cervical instability in middle-aged and young adults.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21292
BACKGROUND: Numerous studies have indicated that stroboscopic visual interference combined with balance training can promote postural stability by reducing visual compensation in the central nervous system and increasing residual proprioceptive and vestibular input during training. OBJECTIVE: To clarify the effects of stroboscopic visual interference combined with balance training on improving balance ability of older adults by comparing the effects of balance training under different visual conditions. METHODS: Forty-three older adults were recruited and randomized into a normal balance training group (n=23) and a strobe vision training group (n=20). Among them, the balance training content was the same, and the training was conducted 3 times per week for 8 weeks. For the strobe vision training group, strobe glasses were worn during the balance training, and the strobe difficulty level (levels 1-8) was adjusted adaptively. Indicator tests were conducted at weeks 0, 4, 8, and 10 to assess the dynamic and static postural stability and the scores on the Berg Balance Scale of all subjects. RESULTS AND CONCLUSION: (1) Static postural stability: In the flat-surface eyes-closed single-leg stance test, both groups showed a significant time main effect (P < 0.001), but the group effect (P=0.530) and group×time interaction effect (P=0.780) were not significant. In the foam-surface eyes-closed single-leg stance test, both groups showed significant time (P < 0.001) and group (P=0.024) effects, but no group×time interaction effect (P=0.063). Compared with before training, both groups showed significant improvements in static postural stability after 8 weeks (P=0.034, P < 0.001) and 10 weeks (P=0.003, P < 0.001). (2) Dynamic postural stability: There was a significant group×time interaction effect (P < 0.001), while the main effects of group (Timed Up and Go and 3-m heel-to-toe walk: P=0.461, P=0.926) and time (P=0.120, P=0.937) were not statistically significant. Compared with before training, both groups showed significant improvements in dynamic postural stability after 4, 8, and 10 weeks of training (all P < 0.05). (3) Berg Balance Scale scores: There was a significant time effect (P < 0.001); group (P=0.420) and group×time (P=0.239) factors had no statistical significance on Berg Balance Scale scores. Compared with before training, both groups had higher Berg Balance Scale scores after 4 weeks (P=0.025), 8 weeks (P < 0.001), and 10 weeks (P=0.003). (4) It is suggested that both traditional balance training and strobe training can significantly improve the dynamic and static stability of older adults. Compared with traditional balance training, balance training based on stroboscopic visual interference can significantly improve the dynamic balance ability of older adults.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21396
BACKGROUND: Current finite element models of the wrist joint predominantly focus on osseous and ligamentous structures, with insufficient incorporation of musculotendinous components, thereby limiting their fidelity and accuracy. OBJECTIVE: To establish a high-fidelity finite element model of the wrist joint, providing a reference for in-depth biomechanical investigations. METHODS: Upper limb CT and MRI data from a 33-year-old healthy male volunteer were imported into Mimics 20.0. Threshold-based selection, region growing, and image segmentation techniques were employed to reconstruct wrist-related bones and soft tissues (including muscles). The model underwent surface optimization, patch generation, and meshing in SolidWorks 2020 and HyperMesh 14.0. Material property assignment and ligament-cartilage contact interfaces were implemented in ABAQUS 6.13 to construct a three-dimensional finite element model of the wrist joint. Stress distribution across wrist structures under axial compression was analyzed. RESULTS AND CONCLUSION: (1) A three-dimensional finite element model encompassing the ulna, radius, distal humerus, carpal bones, metacarpals, pronator teres, pronator quadratus, supinator, lateral muscle group, volar muscle group, dorsal muscle group, interosseous membrane, major ligaments, and cartilage structures was successfully established, comprising 759 191 elements and 245 510 nodes. The stress distribution pattern at the radiocarpal joint under axial compression was obtained and compared with cadaveric studies from the literature, validating the model's authenticity and effectiveness. (2) In summary, based on human CT and MRI imaging data, a more complete wrist joint bone and soft tissue structure was reconstructed through computer software simulation, establishing the origin and insertion points of forearm muscles and their contact with bones during muscle course, resulting in a more realistic finite element model of the wrist joint.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21382
BACKGROUND: Early prediction of cervical instability is crucial for the prevention and treatment of cervical spondylosis, and deep learning technology can provide robust support for intelligent prediction of cervical instability. OBJECTIVE: To develop a deep learning model of cervical instability based on cervical magnetic resonance imaging for early intelligent prediction of cervical instability. METHODS: This study recruited young and middle-aged participants (18-45 years), including both cervical instability patients and healthy controls, through the Spine Department Outpatient Clinic of Wangjing Hospital, China Academy of Chinese Medical Sciences, as well as community-based recruitment. All participants underwent cervical magnetic resonance imaging examinations. On the axial magnetic resonance imaging images, five key anatomical structures were manually annotated: intervertebral disc, facet, prevertebral muscle, deep muscle group in the back of the neck, and superficial muscle group in the back of the neck. A deep learning algorithm was then employed to develop a predictive model for cervical instability, utilizing both the original images and the delineated regions of interest. Finally, the model's predictive performance was systematically evaluated and validated. RESULTS AND CONCLUSION: (1) The study included a total of 308 young and middle-aged participants, comprising 196 individuals with cervical instability and 112 healthy controls. Based on enrollment time, the subjects' data were allocated to either the model training set or the test set. (2) The model demonstrated high predictive performance, with an area under the curve values of 0.97, an F1-score of 0.98, a precision of 0.98, and a recall of 0.97 in the training set. In the test set, these values were 0.97, 0.95, 1.00, and 0.90, respectively. (3) The results indicate that the deep learning model based on cervical magnetic resonance images can achieve early intelligent prediction of cervical instability with high predictive performance.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21352
BACKGROUND: In recent years, with the continuous maturity of the research system, metabolic dysfunction-related fatty liver disease has become independent from traditional non-alcoholic fatty liver disease. Its metabolic disorder background and heterogeneous disease progression patterns have updated the academic understanding of this type of disease. However, the relationship between the common occurrence mechanism of this type of disease and the pathological differences between individuals still needs to be further elucidated through systematic research. OBJECTIVE: To review the common pathways (such as insulin resistance and oxidative stress) commonly found in the pathological mechanism of metabolic dysfunction-related fatty liver disease, and to deeply explore its heterogeneous regulatory network (such as genetic variation and adipose tissue dysfunction), so as to analyze the interaction between the two. METHODS: A systematic search was conducted in Web of Science, PubMed, Embase, CNKI, Wanfang, and VIP databases for Chinese and English literature, with the search time limit from the establishment of each database to June 2025, focusing on the common metabolic disorder mechanisms, genetic/microenvironment heterogeneity pathways, and clinical phenotype classification of metabolic dysfunction-related fatty liver disease, sorting out relevant literature and integrating research evidence. RESULTS AND CONCLUSION: The pathological mechanism of metabolic dysfunction-related fatty liver disease revolves around the core of 'common pathways, heterogeneous regulation, dynamic interaction'. Among the common pathways, insulin resistance is the core link, which activates de novo lipogenesis in the liver, inhibits fatty acid oxidation, and jointly leads to abnormal lipid deposition in hepatocytes; activates the nuclear factor kappa B inflammatory pathway to aggravate hepatocyte injury, and upregulates the transforming growth factor beta pathway to promote liver fibrosis. The synergistic effects of oxidative stress and redox imbalance, and excessive fatty acid accumulation impair mitochondrial function, increase reactive oxygen species production, and destroy cell structure, while an imbalanced state (such as abnormal beta-hydroxybutyrate/acetoacetate ratio) further aggravates injury and promotes the progression of metabolic dysfunction-related fatty liver disease. In terms of heterogeneous regulation, PNPLA3 I148M inhibits triglyceride hydrolysis, TM6SF2 E167K reduces very low-density lipoprotein precursor secretion, independently driving the risk of metabolic dysfunction-related fatty liver disease, liver fibrosis, and cancer; adipose tissue dysfunction is key in lean metabolic dysfunction-related fatty liver disease, leading to ectopic fat deposition and decreased adiponectin levels; among the three metabolic subtypes, type A has lower cardiovascular risk, while types B/C progress rapidly in liver fibrosis. Therefore, the dynamic interaction between genetics, metabolism, and environment affects the disease trajectory, and differentiated intervention based on metabolic subtypes and genetic metabolic risk scores can provide theoretical support for precise risk stratification and personalized treatment of metabolic dysfunction-related fatty liver disease.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21358
BACKGROUND: Pulmonary fibrosis is a chronic progressive lung disease characterized by abnormal deposition of extracellular matrix, with current therapeutic options remaining limited. Extracellular vesicles derived from mesenchymal stem cells, with their lipid membrane structure, can cross the internal barriers in the body and directly deliver various anti-fibrotic, immunomodulatory factors (such as growth factors, immunomodulatory cytokines, and chemokines), lipids and nucleic acids (mRNAs and miRNAs) and other bioactive substances to target cells in the lungs. OBJECTIVE: To systematically review the core mechanisms of mesenchymal stem cell extracellular vesicles in the treatment of pulmonary fibrosis, summarize and elaborate on how extracellular vesicles directly deliver the bioactive substances they carry to different target cells in the lungs, demonstrating their unique advantages in regulating the pulmonary fibrosis microenvironment, and provide a theoretical basis for future use of mesenchymal stem cell-derived extracellular vesicles in the treatment of pulmonary fibrosis. METHODS: A computer-based search was conducted in CNKI, PubMed, clinicaltrials.gov, and the Chinese Clinical Trial Registry. English search terms included "Mesenchymal stem cells, Extracellular vesicles, Pulmonary fibrosis, Alveolar epithelium, Microvascular endothelium, Macrophages, Neutrophils"; Chinese search terms included "间充质干细胞，细胞外囊泡，肺纤维化，上皮细胞，血管内皮细胞，巨噬细胞，中性粒细胞". A total of 56 articles were included for summary. RESULTS AND CONCLUSION: In alveolar epithelial cells, epithelial-mesenchymal transition is inhibited by regulating signaling pathways such as protein kinase B/glycogen synthase kinase 3β and transforming growth factor β/Smad, and specific miRNAs (e.g., miR-466f-3p, let-7) block pro-fibrotic pathway networks. In fibroblasts and endothelial cells, miR-21-5p and miR-218/miR-214-3p respectively interfere with fibroblast activation and endothelial-mesenchymal transition. Meanwhile, they reprogram monocytes, regulate macrophage polarization, inhibit dendritic cell maturation, and balance Th17/Treg responses, reshaping the immune microenvironment. Furthermore, engineered modifications (targeting peptide modification, drug co-loading) of mesenchymal stem cell-derived extracellular vesicles can enhance precise targeting of multiple cell subtypes in diseased areas, while vesicle heterogeneity, standardized production, and in vivo dynamic tracing remain key bottlenecks for clinical translation. Future research should combine single-cell sequencing and spatial multi-omics to deeply analyze the mechanisms of mesenchymal stem cell-derived extracellular vesicles in intervening in cell regulatory networks and develop novel cell-targeted clinical strategies for pulmonary fibrosis.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21527
BACKGROUND: Prolonged forward head posture induces neck muscle fatigue, a significant contributing factor to cervical spondylosis. Current unimodal monitoring approaches are inadequate to capture the dynamic coupling among muscle activation, metabolic activity, and motor control. OBJECTIVE: To systematically evaluate the temporal characteristics of neck muscle fatigue using multimodal monitoring technology, thereby providing a theoretical foundation for early detection and intervention of cervical fatigue. METHODS: Twenty healthy participants were recruited. Surface electromyography, near-infrared spectroscopy, and three-dimensional motion capture technology were synchronized to record electrophysiological signals, oxygenated hemoglobin concentration, and cervical kinematics during a sustained 45° static forward flexion task until subjective fatigue was reached (Borg CR-10 score ≥ 4). Temporal changes in root mean square amplitude, mean power frequency, muscle oxygen saturation, and normalized forward head angle were analyzed across fatigue stages segmented into 10% intervals of total endurance time. RESULTS AND CONCLUSION: (1) The root mean square amplitude increased significantly (P < 0.001), while mean power frequency and muscle oxygen saturation decreased significantly (P < 0.001) throughout the task, with the reduction in muscle oxygen saturation commencing from the 40% fatigue stage. (2) Linear regression analysis between mean power frequency and muscle oxygen saturation showed high explanatory power (upper trapezius R²=0.58, middle trapezius R²=0.61), and metabolic compensation preceded significant electrophysiological changes. (3) The upper trapezius entered fatigue earlier than the middle trapezius (P < 0.05). These results indicate that combined monitoring of mean power frequency and muscle oxygen saturation provides highly sensitive indicators for early warning of neck muscle fatigue.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21583
BACKGROUND: Hydrogel cardiac patches, with their excellent biocompatibility and tunable mechanical properties, demonstrate significant potential in treating acute myocardial infarction, especially when combined with stem cell technology. Hydrogels modified with active components of traditional Chinese medicine (TCM) can promote the proliferation, differentiation, migration, and homing of stem cells. This synergistic effect provides a new approach for stem cell transplantation therapy based on hydrogel cardiac patches.
OBJECTIVE: To focus on the application scenarios of novel biomaterial hydrogel cardiac patches combined with TCM in the treatment of myocardial infarction, and to discuss their development prospects.
METHODS: PubMed and CNKI databases were searched for literature on TCM combined with hydrogel cardiac patches for the treatment of acute myocardial infarction from January 2010 to January 2025. English search terms included "hydrogel, cardiac patch, myocardial infarction, Chinese medicine, stem cell, drug delivery system"; Chinese search terms included "水凝胶, 心脏贴片, 心肌梗死, 中药, 干细胞, 递药系统". According to inclusion and exclusion criteria, 99 articles were finally included for review.
RESULTS AND CONCLUSION: Hydrogel cardiac patches, with their excellent biocompatibility, tunable mechanical properties, and drug-loading capacity, provide ideal mechanical support and microenvironment for myocardial repair. As a hydrophilic polymer biomaterial, the three-dimensional network structure of hydrogel cardiac patches can highly mimic the physical properties of the natural cardiac extracellular matrix, providing a microenvironment for loaded biological cells to proliferate or maintain activity, and helping cell migration, adhesion, spreading, differentiation, and intercellular connection formation. Hydrogels modified with TCM can load stem cells, fully induce and regulate them, and more effectively perform cell regeneration therapy in the myocardial infarction area to achieve better repair. Therefore, the combined application of TCM and hydrogel cardiac patches has great development potential and prospects. Currently, the combined application mainly focuses on the following aspects: precise sustained-release therapy of TCM active ingredients via hydrogel cardiac patch loading; cell regeneration therapy via induced pluripotent stem cells delivered by TCM-modified hydrogel patches; and repair of cardiac electrophysiological function via TCM combined with conductive hydrogels. In future research, it is necessary to deeply understand the characteristics of hydrogels made from different materials and methods, and the therapeutic effects and mechanisms of TCM monomers and compound extracts, to achieve more combined applications of TCM and hydrogel cardiac patches.
Stem Cell Research & Therapy•2026•DOI: 10.1186/s13287-026-04986-7
Tonsil mesenchymal stem cells (TMSCs) represent a promising source for regenerative medicine, yet their clinical translation is constrained by replicative senescence during in vitro expansion. This study investigated senescence-associated phenotypic changes and underlying regulatory mechanisms in human palatine tonsil-derived MSCs. TMSCs were isolated and characterized, then serially passaged to early (P1–P5) and late (beyond P10) stages. Proliferation declined progressively with passage, as assessed by CCK-8. Senescence-associated β-galactosidase (SA-β-gal) staining revealed a significantly higher percentage of positive cells in late-passage TMSCs. Protein levels of p16, p53, and p21 were markedly upregulated in aged cells. RNA sequencing identified differentially expressed genes (DEGs) between young and senescent TMSCs, with KEGG enrichment highlighting the PI3K-Akt signaling pathway, ECM-receptor interaction, and calcium signaling. Western blot confirmed a significantly increased p-Akt/Akt ratio in senescent TMSCs. These findings establish that replicative senescence in TMSCs is associated with activation of the PI3K-Akt pathway, which likely orchestrates senescence through p16 and p53-p21 cascades. The results provide mechanistic insights into stem cell aging and suggest potential molecular targets for delaying TMSC senescence in regenerative applications.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2026048
Pancreatic ductal adenocarcinoma (PDAC) is a malignancy with increasing mortality rates and remains a major clinical challenge due to its aggressive progression and limited therapeutic options. Therefore, the identification of early biomarkers and the development of effective targeted therapies are critically needed. MerTK, a receptor tyrosine kinase aberrantly expressed in various cancers, can be selectively inhibited by UNC569, a small-molecule antagonist with demonstrated efficacy in hematologic malignancies. This study shows that UNC569 potently suppresses PDAC cell proliferation and clonogenic growth, inhibits migration and invasion by attenuating epithelial-mesenchymal transition, and enhances the sensitivity of PDAC cells to Gemcitabine while promoting apoptosis. Mechanistically, UNC569 induces DNA damage-mediated G2/M phase arrest and activates JNK/p38 mitogen-activated protein kinase-dependent apoptotic signaling. Collectively, these results establish MerTK as a promising therapeutic target in PDAC and highlight the translational potential of UNC569 as a dual-pathway inhibitor for PDAC treatment.
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.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2025213
Myocardial infarction (MI) causes irreversible cardiomyocyte loss, and current reperfusion therapies fail to regenerate necrotic myocardium. Cell division cycle 5-like (CDC5L), a cell cycle regulator, has an undefined role in cardiac repair. This study investigates CDC5L in mitigating ischemia-reperfusion (I/R) injury by assessing cardiomyocyte proliferation and apoptosis, and delineates the FGF10-YAP mechanism. In vitro oxygen-glucose deprivation/reoxygenation (OGD/R) in neonatal mouse cardiomyocytes and in vivo I/R in adult mice were employed. CDC5L was modulated via adenoviral or AAV9-mediated overexpression or knockdown. Proliferation markers (EdU+, Ki67+, pH3+), apoptosis (TUNEL, Bax/Bcl-2 ratio), and cardiac function (echocardiography) were quantified. Transcriptomic screening identified downstream targets, validated by FGF10 knockdown rescue. CDC5L was upregulated in post-I/R murine myocardium. Overexpression enhanced cardiomyocyte proliferation, preserved cardiac function, reduced apoptosis, and diminished infarct size. FGF10 was identified as a key downstream effector; CDC5L upregulated FGF10 expression. FGF10 knockdown reversed the proliferative and anti-apoptotic effects of CDC5L. The CDC5L-mediated reduction in YAP phosphorylation was abolished upon FGF10 knockdown. CDC5L attenuates cardiac I/R injury by promoting cardiomyocyte proliferation and inhibiting apoptosis through the FGF10-YAP pathway. This CDC5L-FGF10-YAP axis represents a promising therapeutic target to improve myocardial regeneration and recovery after myocardial infarction.