Stem Cell Research & Therapy•2026•DOI: 10.1186/s13287-026-04896-8
Background The development of vascular calcification (VC) in diabetes is closely related to the endothelial-to-mesenchymal transition (EndMT). We found that microRNA-32-5p (miR-32) was elevated in the plasma of calcification patients. However, it is unclear whether miR-32 mediates the function of bone marrow mesenchymal stem cell-derived extracellular vesicles (BMSC-EVs) in type 2 diabetes (T2D) VC. Methods BMSC-EVs were characterized by TEM, NTA, Western blotting, and confocal microscopy. Alizarin Red and ALP staining assessed the severity of VC. qRT-PCR and Western blotting evaluated the expression of BMP2, RUNX2, GPX4, SLC7A11, VE-cadherin, and N-cadherin, while immunofluorescence was used for detecting VE-cadherin and N-cadherin. In vivo validation was performed using miR-32–/– and ApoE–/– mice. RNA sequencing (RNA-seq) and bioinformatics analysis was conducted to explore underlying mechanisms. Results We demonstrated that BMSC-EVs attenuate VC in endothelial cells (ECs) and inhibit EndMT. In vivo, histological analysis showed that treatment with BMSC-EVs significantly reduced the severity of VC associated with T2D. Notably, knockout of miR-32 further enhanced the inhibitory effect of BMSC-EVs on VC. Mechanistically, transcriptomic and functional analyses suggest that the protective effect of BMSC-EVs on VC is associated with regulation of the MAPK/FoxO signaling pathway, potentially mediated by modulation of ferroptosis. Conclusion These findings demonstrate that BMSC-EVs attenuate T2D-associated VC, partially through miR-32-mediated suppression of EC ferroptosis.
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/s13287-025-04267-9
Background: In premature newborn infants, preterm white matter injury (PWMI) causes motor and cognitive disabilities. Accumulating evidence suggests that PWMI may result from defected differentiation of oligodendrocyte precursor cells (OPCs) and impaired maturation of oligodendrocytes. However, the underlying mechanisms remain unclear. Methods: Using RNAscope, we analyzed the expression level of RNA-binding protein LIN28A in individual OPCs. Knockout of one or both alleles of Lin28a in OPCs was achieved by administrating tamoxifen to NG2CreER::Ai14::Lin28aflox/+ or NG2CreER::Ai14::Lin28aflox/flox mice. Lentivirus expressing FLEX-Lin28a was used in NG2CreER mice to overexpress LIN28A in OPCs. A series of behavioral tests were performed to assess the cognitive functions of mice. Two-tailed unpaired t-tests was carried out for statistical analysis between groups. Results: We found that the expression of Lin28a was decreased in OPCs in a PWMI mouse model. Knockout of one or both alleles of Lin28a in OPCs postnatally resulted in reduced OPC differentiation, decreased myelinogenesis and impaired cognitive functions. Supplementing LIN28A in OPCs postnatally was able to promote OPC differentiation and enhance myelinogenesis, thus rescuing the cognitive functions in PWMI mice. Conclusion: Our study reveals that LIN28A is critical in regulating postnatal myelinogenesis. Overexpression of LIN28A in OPCs rescues cognitive deficits in PWMI mice by promoting myelinogenesis, thus providing a potential strategy for the treatment of PWMI.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2025091
One of the characteristics of malignant tumors is heterogeneity, which refers to the molecular or genetic differences among progeny cells during tumor growth. This heterogeneity contributes to variations in the tumor growth rate, invasive ability, drug sensitivity, and prognosis. To gain a deeper understanding of the molecular background underlying tumor heterogeneity, we construct monoclonal cell lines derived from the glioblastoma (GBM) cell line U87-MG by limiting dilution assays. The selected CF5 and G11 subclones exhibit completely different cell morphologies and, more importantly, distinct functional phenotypes. CF5 exhibits stronger proliferative properties and chemoresistance, whereas G11 shows greater motility and invasion. Transcriptomic sequencing reveals great differences in gene expression among the CF5, G11, and U87 cell lines, and downregulated genes in individual clones are significantly enriched in gene sets related to extracellular matrix function. ITGA11 and ITGA6, as research subjects, are demonstrated to exclusively regulate functional phenotypes and chemotherapy sensitivity in CF5 or G11 cells. In U87 cells, combined knockdown of these two genes significantly inhibits tumor growth and increases chemotherapy sensitivity, but knockdown of either gene alone does not. In summary, these data reveal that even under uniform growth conditions, the heterogeneity of tumor cells and their diverse genetic backgrounds remain significant and persistent. This finding is crucial for accurately identifying tumor-related genes and their functional phenotypes, and a thorough understanding of the genetic and molecular background underlying tumor heterogeneity is essential for comprehensive cancer treatment.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2025140
ATRX is a large, multi-domain nuclear protein that functions as a crucial ATP-dependent chromatin remodeler, transcriptional regulator, and guardian of telomeric and genomic integrity. As a member of the SWI/SNF family of chromatin remodeling proteins, a primary and well-defined function of ATRX is to facilitate the replication-independent deposition of the histone variant H3.3 at specific genomic loci, predominantly repetitive sequences such as telomeres, pericentromeric heterochromatin, and ribosomal DNA (rDNA). Mutations in the ATRX gene are associated with a severe X-linked neurodevelopmental disorder and alpha-thalassemia. Moreover, ATRX dysfunction can lead to genomic instability, contributing to the development and progression of various cancers, including gliomas and pancreatic neuroendocrine tumors (PanNETs). ATRX orchestrates chromatin dynamics through its modular domains. Its N-terminal ADD domain and a PxVxL-like motif recognize histone H3K9me3 and interact with the heterochromatin-binding protein HP1α, respectively. Collaborating with the histone chaperone DAXX, ATRX then utilizes its C-terminal ATPase/Helicase domain to provide the energy needed to remodel chromatin and deposit the histone variant H3.3 into repetitive DNA regions. The Chen lab and other two groups previously elucidated the minimal elements for DAXX interaction, demonstrating that a short 1260‒1289 residues motif (DAXX-binding motif, DBM) of ATRX is solely responsible for ATRX-DAXX heterodimer formation. Beyond its role in chromatin remodeling, ATRX employs its RBR (RNA-binding region) to engage the telomeric long non-coding RNA (lncRNA) TERRA, thereby regulating TERRA-mediated R-loops and telomeric G-quadruplex (G4) structures. Interestingly, this same RBR also binds the muscle-specific lncRNA ChRO1 to coordinate constitutive heterochromatin reorganization and regulate cell differentiation. Notably, nearly half of the disease-causing mutations in ATRX, leading to a severe neurodevelopmental disorder, are clustered within the ADD domain, highlighting this relatively small domain’s critical functional importance and warranting more intense investigation. This Research Highlight discusses recent findings by Yan et al. that the histone variant macroH2A binds the ATRX ADD domain, expanding the known binding partners of this domain and providing structural insights into the interaction.
Acta Biochimica et Biophysica Sinica•2024•DOI: 10.3724/abbs.2024077
Epigenetic modifications play an important role in cellular senescence, and enhancer of zeste homolog 2 (EZH2) is a key methyltransferase involved in epigenetic remodeling in multiple myeloma (MM) cells. We have previously demonstrated that GSK126, a specific EZH2 inhibitor, exhibits anti-MM therapeutic efficacy and safety in vivo and in vitro; however, its specific mechanism remains unclear. This study shows that GSK126 induces cellular senescence in MM, which is characterized by the accumulation of senescence-associated heterochromatin foci (SAHF) and p21, and increased senescence-associated β galactosidase activity. Furthermore, EZH2 is inhibited in ribonucleotide reductase regulatory subunit M2 (RRM2)-overexpressing OCI-MY5 and RPMI-8226 cells. RRM2 overexpression inhibits the methyltransferase function of EZH2 and promotes its degradation through the ubiquitin-proteasome pathway, thereby inducing cellular senescence. In this senescence model, Lamin B1, a key component of the nuclear envelope and a marker of senescence, does not decrease but instead undergoes aberrant accumulation. Meanwhile, phosphorylation of extracellular signal-regulated protein kinase (ERK1/2) is significantly increased. The inhibition of ERK1/2 phosphorylation in turn partially restores Lamin B1 level and alleviates senescence. These findings suggest that EZH2 inhibition increases Lamin B1 level and induces senescence by promoting ERK1/2 phosphorylation. These data indicate that EZH2 plays an important role in MM cellular senescence and provide insights into the relationships among Lamin B1, p-ERK1/2, and cellular senescence.
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.2024199
The renin-angiotensin system (RAS) is a crucial regulatory mechanism for cardiovascular function. The angiotensin II (Ang II) type 1 receptor (AT1R) is the principal receptor responsible for mediating RAS function. AT1R belongs to the G protein-coupled receptor (GPCR) family and is present in multiple tissues, including vascular smooth muscle, endothelium, heart, brain, kidney, adrenal gland, and adipose tissue. Physiologically, AT1R mediates second messenger signaling through classical G proteins. Ang II binding to AT1R predominantly activates Gq/11, leading to the activation of phospholipase C (PLC), which results in the production of inositol 1,4,5-trisphosphate (IP3) and diacylglycerol (DG). Then, increased Ca2+ is released from the sarcoplasmic reticulum to mediate the processes of vasoconstriction, enhance cardiac contractility, regulate water‒salt balance, etc. Under pathological conditions, AT1R aberrantly activates G proteins, including mitogen-activated protein kinases (MAPKs: ERK1/2, JNK, and p38MAPK), receptor tyrosine kinases (PDGF, EGFR, and insulin receptor), non-receptor tyrosine kinases [Src, JAK/STAT, and focal adhesion kinase (FAK)], and NADPH oxidase, to influence downstream pathways. This activation exacerbates inflammatory responses, fibrosis, and pathological cardiovascular remodeling. AT1Rs within the nervous system can also induce excessive activity in the sympathetic nervous system, which increases myocardial strain and facilitates the progression of heart failure. Owing to the importance of AT1R in a variety of diseases, greater demands have been placed on the accuracy of AT1R detection. The structural complexity and low immunogenicity of GPCRs pose considerable challenges in the development of specific antibodies. Many commercially available antibodies for GPCRs, such as those against muscarinic and adrenergic receptors, lack specificity. Current studies on AT1R often use these commercial antibodies, but many fail to demonstrate specificity when AT1R-knockdown or AT1R-overexpressing tissues and cells are tested. This study aims to specifically validate six newly available commercial AT1R antibodies (Supplementary Table S1). Using AT1R global knockout SD rats, cardiomyocyte conditional AT1R knockout C57BL/6N mice, AT1R-overexpressing CHO stable-transformed cell lines and AT1R-overexpressing HEK293 cells, we assessed AT1R expression and localization through receptor-ligand binding assays, RT-PCR, western blot analysis, and immunocytochemistry. Materials and methods are available in Supplementary Materials and Methods. To verify the specificity of the antibody, we generated AT1R-global knockout SD rats (AT1R-KO) using CRISPR-Cas9 technology. Agarose gel electrophoresis revealed bands at approximately 470 bp for AT1R-KO rats and 531 bp for wild-type (WT) rats, confirming successful AT1R knockout at the gene level (Figure 1A). RT-PCR analysis of vascular tissue RNA revealed the absence of AT1R in AT1R-KO rats (Figure 1B). Ligand-receptor binding assays revealed significantly less 125I-Ang II binding to vascular tissue proteins in AT1R-KO rats than in WT rats (Figure 1C). Additionally, primary cardiomyocytes extracted from 0–3-day-old WT and AT1R-KO neonatal rats presented a significant increase in beating rate upon Ang II stimulation in WT rats, whereas no response was observed in AT1R-KO rats (Figure 1D). These results confirmed successful AT1R global knockout in AT1R-KO rats. AT1R-KO rats were thus utilized to verify the specificity of AT1R antibodies (A14201, 25343-1-AP, and 66415-1-Ig). Western blot analysis was conducted on protein extracts from the heart, vascular, liver, and kidney tissues of WT and AT1R-KO rats. Under room temperature denaturation conditions, the A14201 antibody detected AT1R bands at the expected molecular weight (42 kDa) in all tissues from WT rats, the 25343-1-AP antibody detected AT1R in heart and kidney tissues, and the 66415-1-Ig antibody detected AT1R only in the heart tissues. Compared with WT control rats, the A14201 antibody revealed a reduction in AT1R protein expression in AT1R-KO rats.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2024194
Glucocorticoids (GCs) are commonly used to treat sudden sensorineural hearing loss (SSNHL), although some patients are resistant to this therapeutic approach. Clinical studies have demonstrated the efficacy of tanshinone IIA (TA) in combination with GC for managing various human ailments. However, it remains unclear whether TA can mitigate GC resistance in SSNHL. Our aim is to elucidate the role of NRF2-induced transcriptional regulation of HDAC2 in influencing GC resistance and investigate the involvement of TA-related molecular pathways in GC resistance. Here, HEI-OC1 cells are treated with lipopolysaccharide (LPS) to establish an in vitro model for SSNHL. The cells are subsequently treated with dexamethasone (DXE) or DXE + TA. RT-qPCR and western blot analysis are used to measure the mRNA and protein levels of Forkhead box P3 (FOXP3), nuclear factor erythroid 2-related factor 2 (NRF2), and histone deacetylase 2 (HDAC2). Cell Counting Kit-8 (CCK-8) and 5-ethynyl-2’-deoxyuridine (EdU) assays are carried out to assess cell proliferation. Flow cytometry analysis is performed to evaluate apoptosis. Mechanistic studies involve chromatin immunoprecipitation (ChIP), luciferase reporter, and DNA pull-down assays. Our results show that treatment with TA + DEX significantly increases proliferation and suppresses apoptosis in LPS-treated HEI-treated OC1 cells. TA upregulates HDAC2 expression by activating NRF2-mediated transcription of HDAC2, with the NRF2-HDAC2 binding site located at bases 419–429 (ATGACACTCCA) in the promoter sequence of HDAC2. Furthermore, TA upregulates FOXP3 expression to activate NRF2 transcription, with the predicted FOXP3-binding site located at bases 864–870 (GCAAACA) in the promoter sequence of NRF2. In summary, these findings suggest that TA enhances the therapeutic effects of GC on the proliferation and apoptosis of HEI OC1 cells by increasing FOXP3/Nrf2 expression. These results indicate that TA may be promising for ameliorating GC resistance in patients with SSNHL.
Acta Biochimica et Biophysica Sinica•2024•DOI: 10.3724/abbs.2024224
ADP-ribosylation factor collaborator (CARF), which is also known as CDKN2AIP, was first recognized as an ADP-ribosylation factor-interacting protein that participates in the activation of the ARF-p53-p21 (WAF1) signaling pathway under different conditions, such as oxidative and oncogenic stresses. The activation of this pathway often leads to cell growth arrest and apoptosis as well as senescence. Previous studies revealed that CARF, an RNA-binding protein, is critical for maintaining stem cell pluripotency and somatic differentiation. Nevertheless, its involvement in spermatogenesis has not been well examined. In this study, we show that male mice deficient in Carf expression present impaired spermatogenesis and fertility. IP-MS and RNA-seq analyses reveal that CARF/Carf interacts with multiple key splicing factors, such as PABPC1, and directly targets 356 different types of mRNAs in spermatocytes. Carf-associated mRNAs display aberrant splicing patterns when Carf expression is deficient. In addition, our results demonstrate that PIWIL1 expression and localization are altered in the Carf-/- mouse model through the downregulation of PABPC1, which further affects the ratio of pachytene-piRNA. Our study suggests that CARF is critical for regulating alternative splicing in mammalian spermatogenesis and determining infertility in male mice.
Stem Cell Research & Therapy•2026•DOI: 10.1186/s13287-026-04896-8
Vascular calcification (VC) in type 2 diabetes (T2D) is driven by endothelial-to-mesenchymal transition (EndMT), yet effective therapies remain elusive. Elevated plasma microRNA-32-5p (miR-32) correlates with calcification, but its role in bone marrow mesenchymal stem cell-derived extracellular vesicle (BMSC-EV) therapy is undefined. We characterized BMSC-EVs by TEM, NTA, Western blotting, and confocal microscopy. Alizarin Red and ALP staining quantified VC severity. qRT-PCR and Western blotting assessed BMP2, RUNX2, GPX4, SLC7A11, VE-cadherin, and N-cadherin; immunofluorescence localized VE-cadherin and N-cadherin. In vivo validation used miR-32–/– and ApoE–/– mice. RNA sequencing and bioinformatics explored mechanisms. BMSC-EVs attenuated VC in endothelial cells (ECs) and inhibited EndMT. In vivo, BMSC-EV treatment significantly reduced T2D-associated VC severity. Notably, miR-32 knockout further enhanced the inhibitory effect of BMSC-EVs on VC. Transcriptomic and functional analyses linked the protective effect to MAPK/FoxO signaling modulation, potentially via ferroptosis regulation. These findings demonstrate that BMSC-EVs attenuate T2D-associated VC, partially through miR-32-mediated suppression of EC ferroptosis, providing a mechanistic foundation for EV-based therapeutics.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21244
BACKGROUND: Studies have shown that miRNAs, as important post-transcriptional regulators of genes, play a key role in the onset and progression of osteoporosis. Through in-depth research on the biology of miRNA regulation of osteoporosis, its potential healing mechanisms have been revealed, and this field has become a hot focus of current research. OBJECTIVE: To explore the regulatory role of miRNAs in the development of osteoporosis and their molecular mechanisms, and to provide an overview of the key difficulties encountered in the therapeutic strategies for osteoporosis targeting miRNAs and their solutions. METHODS: We searched PubMed, Web of Science and CNKI databases for relevant literature published up to March 2025. The search terms were “miRNA, osteoporosis, angiogenesis, osteogenesis, gene therapy, drug delivery” in English and “miRNA, osteoporosis, gene therapy, ribonucleic acid drugs, delivery carrier” in Chinese. After reading the titles and abstracts for preliminary screening, we excluded the literature with poor relevance, old information, or repetitive views and lack of authority, and finally included 138 papers for review. RESULTS AND CONCLUSION: (1) miRNAs are highly efficient non-coding RNAs with a wide range of applications that can precisely regulate cellular activities, and they show significant therapeutic potential in regulating osteoblast function and bone angiogenesis. (2) Although miRNA-based targeted drugs have entered preclinical research in other disease areas, clinical translation still faces challenges of insufficient nucleic acid stability in vivo and off-target effects. (3) To address these challenges, researchers have proposed various strategies, including precise targeting of miRNA target genes to reduce off-target effects; chemical modification to improve the stability of nucleic acid drugs in vivo; reducing nucleic acid production costs to advance research; and utilizing viral vectors, exosomes, and various biomaterials to optimize nucleic acid drug delivery routes. (4) Advances in technology continue to innovate in improving the performance of nucleic acid drug carriers, and in the future, precise and efficient drug delivery and targeted therapeutic effects will be achieved.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21215
BACKGROUND: Transverse tibial bone transfer is an emerging surgical technique that enhances local blood circulation and promotes angiogenesis, thereby accelerating the healing of diabetic foot ulcers. Although this technique has demonstrated positive clinical outcomes, its specific molecular mechanisms remain unclear. Recently, the role of circular RNA in angiogenesis and wound healing has gained increasing recognition. Circular RNA may influence the healing process by regulating the expression of related genes; however, its involvement in the treatment of diabetic foot ulcers through transverse tibial bone transfer has yet to be explored. OBJECTIVE: To investigate the therapeutic effects of transverse tibial bone transfer on diabetic foot ulcers in a rabbit model and the mechanism of action. METHODS: Eighteen 3-month-old male New Zealand rabbits, weighing 2.8–3.6 kg, were included in this study. After being fed a high-sugar, high-fat diet for 1 month, type II diabetic rabbit models were induced by intravenous injection of alloxan monohydrate. After successful modeling, the right femoral artery at the mid-upper segment was ligated, and full-thickness skin on the ipsilateral foot dorsum was excised to simulate the pathological features of diabetic foot ulcers. Subsequently, the successfully modeled rabbits were randomly divided into 4 groups (4 rabbits per group): blank group (no additional treatment), dressing change group (routine iodophor disinfection after modeling), sham surgery group (installation of transverse tibial bone transfer scaffold without bone transfer), and surgery group (installation of scaffold and bone transfer). At 7 and 14 days post-surgery, the healing of foot ulcer wounds was observed. At 7, 14, and 21 days post-surgery, serum levels of vascular endothelial growth factor A (VEGF-A) and CD31 were measured by enzyme-linked immunosorbent assay. At 14 days post-surgery, ulcer tissue samples were collected for hematoxylin-eosin staining, CD31 immunofluorescence staining, and western blot analysis of VEGF-A and CD31 protein expression. At 7, 14, and 21 days post-surgery, venous blood from the surgery group was collected for whole-genome sequencing to analyze differential expression of circular RNAs. RESULTS AND CONCLUSION: At 7 and 14 days post-surgery, the surgery group showed significantly better recovery of diabetic foot ulcers compared to the other three groups, with superior promotion of epidermal repair, collagen fiber deposition, and angiogenesis. At 14 and 21 days post-surgery, serum levels of VEGF-A and CD31 in the surgery group were significantly higher than those in the other three groups (P < 0.01). Gene sequencing analysis revealed that the most significant changes in circular RNAs occurred at 21 days post-surgery, especially the expression of circular RNA PDS5B (circPDS5B) adhesion-related factor B, which gradually decreased over time, suggesting that circPDS5B may be closely related to angiogenesis and tissue repair. These results indicate that transverse tibial bone transfer can effectively promote the healing of diabetic foot ulcer wounds in rabbits. Gene sequencing results showed differential expression of circular RNAs, especially significant downregulation of circPDS5B, suggesting that transverse tibial bone transfer may promote wound repair and angiogenesis by activating related molecular pathways.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21311
BACKGROUND: Alzheimer’s disease (AD) is a neurodegenerative disorder. Although β-amyloid and Tau proteins are core biomarkers for AD diagnosis, their heterogeneity and diagnostic limitations necessitate the exploration of novel biomarkers for disease diagnosis and treatment. OBJECTIVE: To analyze the interaction between N6-methyladenosine (m6A) epitranscriptomic modifications and ferroptosis genes in AD using machine learning, bioinformatics analysis, and experimental validation, to identify characteristic genes for AD pathogenesis, and to reveal their association with immune microenvironment regulation, thereby providing novel biomarkers for early diagnosis and precise treatment of AD. METHODS: Genomic data of human hippocampal tissues from GSE5281, GSE48350 (training sets), and GSE33000 (validation set) in the GEO database were integrated. Differentially expressed m6A regulators in AD were screened in the training sets, and the correlation between m6A and ferroptosis genes was assessed to identify ferroptosis-related differentially expressed genes associated with m6A. Support vector machine recursive feature elimination combined with Boruta feature selection was used to determine AD characteristic genes. Gene set enrichment analysis was performed to dissect functional modules of characteristic genes. A logistic regression model combined with receiver operating characteristic curves was constructed to evaluate the diagnostic efficacy of characteristic genes in the validation set. Single-sample gene set enrichment analysis was applied to quantify immune cell infiltration levels and analyze their regulatory association with characteristic genes. Transcription factor/miRNA-mRNA regulatory networks were predicted using ENCORI, miRWalk 3.0, and NetworkAnalyst databases. Potential therapeutic compounds were screened via the CTD database. qRT-PCR and western blotting were used to validate characteristic genes in hippocampal tissues of APP/PS1 double-transgenic mice. RESULTS AND CONCLUSION: (1) Two significantly differentially expressed m6A regulators, Wilms tumor 1 associated protein (WTAP) and methyltransferase-like protein 14 (METTL14), were identified, with 16 ferroptosis-related genes associated with them. (2) Machine learning identified five core characteristic genes: fumarate hydratase (FH), aspartate aminotransferase (GOT1), HRas proto-oncogene (HRAS), metallothionein 3 (MT3), and SET domain containing 1B (SETD1B). (3) Characteristic genes were functionally enriched in oxidative phosphorylation, Huntington disease, Parkinson disease, fatty acid degradation and metabolism, and proteasome signaling pathways. (4) The logistic regression diagnostic model achieved area under the curve values of 0.873 and 0.904 in the training and validation sets, respectively, indicating excellent diagnostic efficacy. (5) Immune microenvironment analysis showed that HRAS was significantly correlated with chemokine receptor family and plasmacytoid dendritic cell infiltration levels. (6) A regulatory network comprising 5 mRNAs, 37 miRNAs, and 142 transcription factors was constructed, and 71 potential therapeutic drugs were predicted. (7) Experimental validation showed that mRNA and protein expression of GOT1, HRAS, and SETD1B in the hippocampus of APP/PS1 mice were significantly different (P < 0.05 or P < 0.01), consistent with bioinformatics analysis. (8) The results reveal that FH, GOT1, HRAS, MT3, and SETD1B can serve as characteristic genes for AD; immune infiltration correlation analysis suggests that HRAS may serve as a potential immunotherapeutic marker for AD, providing a theoretical basis for early diagnosis and targeted therapy.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21393
BACKGROUND: The elastic modulus of β-tricalcium phosphate bioceramic rods is close to that of normal bone tissue, and it exhibits excellent biocompatibility and mechanical properties. It can be used as a supporting material inside the femoral head after core decompression. However, there are few biomechanical studies on osteonecrosis of the femoral head and the changes in stress and displacement of the femoral head after ceramic rod implantation. OBJECTIVE: To explore the biomechanical effects of core decompression with ceramic rod implantation in the treatment of osteonecrosis of the femoral head during the peri-collapse stage. METHODS: A total of 21 hips were selected from 19 patients with osteonecrosis of the femoral head implanted with ceramic rods at the peri-collapse stage. Preoperative and postoperative imaging data were obtained, and relevant CT images were loaded in Mimics 21.0 software to construct a three-dimensional model of the femoral head. A global model of the proximal femur that includes cortical and cancellous bone, as well as a model of the proximal cancellous bone of the femur were created. The preoperative MRI image data of the patients were imported, and the necrotic lesion model was made by using the graphic matching technology, which was saved in .stl format. They were transferred to Geomagic 2012 software for smoothing and precise surface processing. Subsequently, the ceramic rod was designed and modeled in SolidWorks 2021 software, and the relevant models were imported for assembly and Boolean operations. After ensuring no interference, ANSYS 2021 software was used to calculate and observe the stress and displacement of the weight-bearing area and necrotic area of the femoral head during single-leg stance and the push-off phase of walking. RESULTS AND CONCLUSION: (1) The area of maximum stress on the femoral head was located in the anterolateral superior part of the necrotic area. During single-leg stance, the stress values in the weight-bearing area and necrotic area were significantly lower postoperatively than preoperatively (P < 0.05), and the femoral head collapse value (displacement of the weight-bearing area) was lower than preoperatively (P < 0.05). (2) During the push-off phase of walking, with the increase in load, the stress values in the weight-bearing area and necrotic area and the femoral head collapse value (displacement of the weight-bearing area) increased, but they were still lower than preoperatively (P < 0.05). (3) It is suggested that core decompression combined with ceramic rod implantation helps to reduce the load on the weight-bearing area of the femoral head, effectively disperse the stress in the weight-bearing area, partially transfer the load to the femoral calcar, improve the local stress concentration, and effectively support the femoral head to prevent further collapse.
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: 10.12307/2026.21522
BACKGROUND: Tibial transverse transport has emerged as a pivotal therapeutic approach for severe diabetic foot ulcers (Wagner grade III and above); however, its precise molecular regulatory mechanisms remain largely elusive. OBJECTIVE: To establish an animal model of tibial transverse transport for diabetic foot ulcer treatment and perform transcriptome sequencing, aimed at identify significantly differentially expressed miRNAs and key target genes, as well as construct the corresponding miRNA-mRNA regulatory network. METHODS: A diabetic foot ulcer wound model was established in rabbits and treated with tibial transverse transport. Peripheral blood samples were collected for miRNA-seq and mRNA-seq. Differentially expressed miRNAs and mRNAs were identified through bioinformatics analysis of sequencing data. Target genes of miRNAs were predicted using TargetScan and miRanda, and a miRNA-mRNA regulatory network was constructed. Kyoto Encyclopedia of Genes and Genomes enrichment analysis and protein-protein interaction analysis were performed on target genes, and core target genes were screened using MCC and Degree algorithms. RESULTS AND CONCLUSION: A total of 9 significantly differentially expressed miRNAs and 2,667 significantly differentially expressed mRNAs were identified. Based on differential analysis and predicted miRNA target genes, a miRNA-mRNA regulatory network containing 7 miRNAs and 79 target genes was constructed. Kyoto Encyclopedia of Genes and Genomes enrichment analysis revealed that these target genes were mainly involved in metabolic pathways, protein processing in endoplasmic reticulum, FoxO signaling pathway, propanoate metabolism, and N-glycan biosynthesis. Protein-protein interaction analysis revealed interactions among the proteins encoded by these target genes and identified BAG3, AKT3, PPP4C, SEC61A1, DNAJC3, USP7, DAD1, and SETD7 as core target genes. These results indicate that a series of key miRNAs, target genes, and signaling pathways related to diabetic foot ulcer treatment were identified through transcriptome sequencing, providing new candidates for therapeutic targets and opening new avenues for exploring the therapeutic mechanism of tibial transverse transport.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21523
BACKGROUND: Recent studies on the pathogenesis of multiple sclerosis suggest that intervening in glial cells may play a key role in reducing relapses and delaying disability progression. Grape seed proanthocyanidin oligomers significantly inhibit demyelination in cuprizone-treated mice. OBJECTIVE: To explore the mechanism by which grape seed proanthocyanidin oligomers protect myelin sheaths through regulating astrocytes. METHODS: (1) Animal experiment: Thirty mice were randomly divided into normal group, CPZ group, and CPZ+oligomeric proanthocyanidin group. The latter two groups were fed a diet containing 0.2% CPZ for 6 weeks to induce demyelination. From the 5th week, the normal and CPZ groups were given ddH2O by gavage, while the CPZ+oligomeric proanthocyanidin group received grape seed proanthocyanidin oligomers [50 mg/(kg·d)] once daily for 2 weeks. Behavioral changes were observed; LFB and oil red staining were used to assess myelin pathology; ELISA detected inflammatory factors in the brain; immunofluorescence staining detected related protein expression. (2) Cell experiment: In vitro, grape seed proanthocyanidin oligomers (30 μg/mL) were used to intervene in an astrocyte inflammation model induced by tumor necrosis factor α, interleukin 1α, and C1q. Conditioned medium was collected and used to culture oligodendrocytes. Cells were divided into normal, model, and model+oligomeric proanthocyanidin groups. L-lactate dehydrogenase and CCK-8 assays were used to detect oligodendrocyte damage and cell activity, and western blot was used to detect apoptosis-related protein expression. RESULTS AND CONCLUSION: (1) Grape seed proanthocyanidin oligomers significantly improved demyelination in CPZ mice, inhibited the expression of pro-inflammatory factors tumor necrosis factor α, interleukin 6, interleukin 1α, and interleukin 17 in the brain, promoted the secretion of anti-inflammatory factor transforming growth factor β, accompanied by astrocyte proliferation in the corpus callosum, significantly reduced the marker C3d of pro-inflammatory astrocytes, and inhibited the phosphorylation of JNK, a signaling molecule related to astrocyte polarization. (2) Compared with the model group, the conditioned medium after intervention with grape seed proanthocyanidin oligomers significantly reduced the apoptosis of oligodendrocytes induced by inflammatory astrocytes, promoted the expression of Bcl-2, and inhibited the expression of Bax and Caspase-3. (3) These results indicate that grape seed proanthocyanidin oligomers can inhibit demyelination in CPZ mice by inhibiting JNK phosphorylation in astrocytes, reducing the polarization of astrocytes to the pro-inflammatory A1 type, thereby inhibiting the apoptosis of oligodendrocytes induced by inflammatory astrocytes.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2025091
Glioblastoma multiforme (GBM) exhibits profound intratumoral heterogeneity that drives therapeutic resistance and recurrence. Using limiting dilution, we derived monoclonal sublines CF5 and G11 from the U87-MG GBM cell line. These subclones displayed divergent morphologies and functional phenotypes: CF5 demonstrated enhanced proliferation and chemoresistance, whereas G11 exhibited increased motility and invasion. Transcriptomic sequencing revealed extensive differential gene expression among CF5, G11, and parental U87 cells, with downregulated genes in individual clones significantly enriched in extracellular matrix (ECM)-related gene sets. ITGA11 and ITGA6 were identified as exclusive regulators of phenotype and chemotherapy sensitivity in CF5 and G11, respectively. In mixed U87 cells, single knockdown of either ITGA11 or ITGA6 failed to produce substantial phenotypic changes, but combined knockdown significantly inhibited tumor growth and increased chemosensitivity. These findings underscore that tumor heterogeneity and diverse genetic backgrounds persist even under uniform culture conditions, obscuring the functional contributions of individual genes in bulk populations. The study highlights the necessity of resolving subclonal expression patterns to accurately assign gene function and to design effective combinatorial targeted therapies. Subcutaneous tumor models, while not fully recapitulating the brain microenvironment, provided practical monitoring of tumor dynamics. This work challenges single-target therapeutic strategies and advocates for context-dependent molecular interventions in GBM.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2025126
Ventricular arrhythmias (VAs) are the predominant cause of mortality following myocardial infarction (MI), driven by electrophysiological remodeling including dysregulated calcium cycling. Prior work demonstrated that CDR1as knockdown ameliorates arrhythmias by modulating Nav1.5 and Kir6.2 channels post-MI. This study investigates the role of CDR1as in calcium channel remodeling after ischemic arrhythmia. MI was induced in mice by left anterior descending coronary artery ligation, and patch-clamp techniques measured Ca current (ICaL) in isolated ventricular cardiomyocytes. Cav1.2 expression was significantly decreased in the infarct border zone at 12 h post-MI. CDR1as knockdown via AAV9-CDR1as-shRNA administration enhanced cardiac function and restored both ICaL density and Cav1.2 expression in MI model mice. The data suggest that MI alters cardiac calcium expression and increases vulnerability to VAs, and that targeting the CDR1as pathway to modulate calcium channels may be a viable antiarrhythmic strategy post-MI. Notably, CDR1as did not significantly affect RyR2 protein, though it regulated SERCA2a expression. The study hypothesizes that CDR1as may regulate Cav1.2 via mRNA methylation or by acting as an endogenous inhibitor of Cav1.2. These findings reveal an unprecedented role for CDR1as in post-MI arrhythmias and support further investigation into its therapeutic potential.