Stem Cell Research & Therapy•2025•DOI: 10.1186/s13287-025-04245-1
Background Chronic limb-threatening ischemia (CLTI) is the most severe form of peripheral arterial disease (PAD). Mesenchymal stem cell (MSC) transplantation holds promise as a treatment for CLTI; however, the harsh local environment poses challenges to its effectiveness. Apoptotic vesicles (ApoVs) are extracellular vesicles produced by cells undergoing apoptosis, and they can carry various biomolecules from their parent cells, including proteins, RNA, DNA, lipids, ions, and gas neurotransmitters. ApoVs play significant roles in anti-inflammatory responses, anti-tumor activities, and tissue regeneration through intercellular communication, and they have demonstrated potential as drug carriers. In this study, we investigated the potential of bone marrow stem cell (BMSC)-derived ApoVs for treating CLTI. Methods In vivo, we explored the therapeutic effect of ApoVs on a hindlimb ischemia model through Laser Doppler, matrigel plug assay, and histological analysis. In vitro, we analyzed the effects of ApoVs on the proliferation, migration, and angiogenesis of HUVECs and explored the uptake process of ApoVs. In addition, Proteomic analysis, western blotting, quantitative real-time PCR, shRNA, and siRNA were used to analyze ApoVs-induced HUVECs activation and downstream signaling pathways. Results BMSCs transplantation showed improvement in a hind limb ischemia model, and this effect still exists after apoptosis of BMSCs. Subsequently, ApoVs of BMSCs were isolated and found to improve mouse hind limb ischemia in vivo. In vitro, ApoVs can be ingested by HUVECs through dynamin-, clathrin-, and caveolin-mediated endocytosis and promote its proliferation, migration, and angiogenesis. Mechanistically, ApoVs transferred NAMPT to HUVECs, therefore activating the NAMPT/SIRT1/FOXO1 axis, influencing the transcriptional activity of FOXO1, and promoting angiogenesis. Conclusions Our results demonstrate that the transplanted BMSCs can ameliorate hindlimb ischemia by releasing ApoVs during apoptosis. The main mechanism of this effect is promoting the proliferation, migration, and angiogenesis of endothelial cells via the NAMPT/SIRT1/FOXO1 axis.
Stem Cell Research & Therapy•2025•DOI: 10.1186/s13287-025-04396-1
Background One major factor contributing to infertility in women of childbearing age is premature ovarian insufficiency (POI). Exosomes produced from human umbilical cord mesenchymal stem cells (hUMSC-Exos) have drawn a lot of attention lately as a potential treatment for ovarian dysfunction brought on by POI. However, its therapeutic mechanism is still unclear and needs further exploration.
Methods POI model was established by intraperitoneal injection of cyclophosphamide (CTX) in female Wistar rats. These POI rats were treated with hUMSC-Exos for one week. In addition to in vivo experiments, in vitro POI models were also established. In vitro experiments, theca interstitial cells (TICs) treated with CTX were exposed to normal as well as let-7a-5p inhibitory hUMSC-Exos. The ovary structure, morphology, endocrine function, and reproductive ability of POI rats were observed by H&E staining and ELISA. Western blot, immunofluorescence staining (IF), and quantitative real-time polymerase chain reaction (qRT-PCR) were used to evaluate the autophagy-related indexes in ovary and TICs of POI rats in each group.
Results CTX induced abnormalities of ovarian morphology, structure, endocrine, and reproductive function in rats, and accompanied by autophagy of TICs. Notably, hUMSC-Exos diminishes ovarian structural and functional damage in POI rats and TICs autophagy via targeting the AMPK/mTOR pathway. Furthermore, downregulating let-7a-5p in hUMSC-Exos weakened their ability to prevent TICs autophagy.
Conclusions Overall, the findings suggested that hUMSC-Exos improves ovarian function in POI rats by inhibiting TICs autophagy via the let-7a-5p/AMPK/mTOR pathway. Our study provided further evidence that POI patients can benefit from hUMSC-Exos-mediated therapy.
Stem Cell Research & Therapy•2025•DOI: 10.1186/s13287-025-04202-y
Background Severe hepatic steatosis can exacerbate Ischemia–reperfusion injury (IRI), potentially leading to early graft dysfunction and primary non-function. In this study, we investigated the heterogeneity of different subpopulations of Urine-derived stem cells (USCs) to explore the most suitable cell subtype for treating severe steatotic liver IRI. Methods This study utilized scRNA-seq and Bulk RNA-seq to investigate the transcriptional heterogeneity between Spindle-shaped USCs (SS-USCs) and Rice-shaped USCs (RS-USCs). Additionally, rat fatty Liver transplantation (LT) model, mouse fatty liver IRI model, and Steatotic Hepatocyte Hypoxia-Reoxygenation (SHP-HR) model were constructed. Extracellular vesicles derived from SS-USCs and RS-USCs were isolated and subjected to mass spectrometry analysis. The therapeutic effects of Spindle-shaped USCs Exosomes (SS-USCs-Exo) and Rice-shaped USCs Exosomes (RS-USCs-Exo) were explored, elucidating their potential mechanisms in inhibiting ferroptosis and alleviating IRI. Results Multiple omics analyses confirmed that SS-USCs possess strong tissue repair and antioxidant capabilities, while RS-USCs have the potential to differentiate towards specific directions such as the kidney, nervous system, and skeletal system, particularly showing great application potential in renal system reconstruction. Further experiments demonstrated in vivo and in vitro models confirming that SS-USCs and SS-USCs-Exo significantly inhibit ferroptosis and alleviate severe fatty liver IRI, whereas the effects of RS-USCs/RS-USCs-Exo are less pronounced. Analysis comparing the proteomic differences between SS-USCs-Exo and RS-USCs-Exo revealed that SS-USCs-Exo primarily inhibit ferroptosis and improve cellular viability by secreting exosomes containing Glutathione Peroxidase 4 (GPX4) protein. This highlights the most suitable cell subtype for treating severe fatty liver IRI. Conclusions SS-USCs possess strong tissue repair and antioxidant capabilities, primarily alleviating ferroptosis in the donor liver of fatty liver through the presence of GPX4 protein in their exosomes. This highlights SS-USCs as the most appropriate cell subtype for treating severe fatty liver IRI.
Stem Cell Research & Therapy•2024•DOI: 10.1186/s13287-023-03572-5
Objective In recent years, cell therapy has emerged as a new research direction in the treatment of diabetes. However, the underlying molecular mechanisms of mesenchymal stem cell (MSC) differentiation necessary to form such treatment have not been clarified. Methods In this study, human umbilical cord mesenchymal stem cells (HUC-MSCs) isolated from newborns were progressively induced into insulin-producing cells (IPCs) using small molecules. HUC-MSC (S0) and four induced stage (S1–S4) samples were prepared. We then performed transcriptome sequencing experiments to obtain the dynamic expression profiles of both mRNAs and long noncoding RNAs (lncRNAs). Results We found that the number of differentially expressed lncRNAs and mRNAs trended downwards during differentiation. Gene Ontology (GO) analysis showed that the target genes of differentially expressed lncRNAs were associated with translation, cell adhesion, and cell connection. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed that the NF-KB signalling pathway, MAPK signalling pathway, HIPPO signalling pathway, PI3K–Akt signalling pathway, and p53 signalling pathway were enriched in these differentially expressed lncRNA-targeting genes. We also found that the coexpression of the lncRNA CTBP1-AS2 with PROX1 and the lncRNAs AC009014.3 and GS1-72M22.1 with JARID2 mRNA was related to the development of pancreatic beta cells. Moreover, the coexpression of the lncRNAs: XLOC_ 050969, LINC00883, XLOC_050981, XLOC_050925, MAP3K14- AS1, RP11-148K1.12, and CTD2020K17.3 with p53, regulated insulin secretion by pancreatic beta cells. Conclusion In this study, HUC-MSCs combined with small molecule compounds were successfully induced into IPCs. Differentially expressed lncRNAs may regulate the insulin secretion of pancreatic beta cells by regulating multiple signalling pathways. The lncRNAs AC009014.3, Gs1-72m21.1, and CTBP1-AS2 may be involved in the development of pancreatic beta cells, and the lncRNAs: XLOC_050969, LINC00883, XLOC_050981, XLOC_050925, MAP3K14-AS1, RP11-148K1.12, and CTD2020K17.3 may be involved in regulating the insulin secretion of pancreatic beta cells, thus providing a lncRNA catalogue for future research regarding the mechanism of the transdifferentiation of HUC-MSCs into IPCs. It also provides a new theoretical basis for the transplantation of insulin-producing cells into diabetic patients in the future.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2026006
Protein homeostasis serves as the foundation for every cellular decision—division, differentiation, stress adaptation, or death—by precisely balancing the proteome across abundance, quality, spatial distribution, and temporal dynamics; its dysregulation drives numerous human pathologies, including cancers and neurological disorders. In the traditional ubiquitin-dependent degradation cascade, target proteins are marked by covalent attachment of polyubiquitin chains, a process requiring E1 activating enzymes, E2 conjugating enzymes, and E3 ligases that confer substrate specificity. This ubiquitin signal is then recognized by the 19S regulatory particle of the proteasome, which unfolds and translocates the tagged protein into the 20S core for proteolytic destruction. The ubiquitin-independent proteasomal degradation pathway mediated by midnolin (MIDN) has recently emerged as a distinct and biologically important mechanism for regulating nuclear protein turnover. While earlier genetic, biochemical, and cryo-electron microscopy studies established MIDN as a proteasome-associated adaptor for immediate-early gene (IEG) products, the molecular logic underlying its broad yet selective substrate recognition remains unresolved. In the current study, Zhong et al. provide a comprehensive structural and biochemical analysis of the MIDN Catch domain bound to IRF4-derived peptides, substantially advancing our understanding of MIDN substrate recognition. By solving multiple crystal structures of the Catch-IRF4 complex, including wild-type and systematically engineered mutants, the authors demonstrate that MIDN recognizes substrates through a conserved β-strand insertion mechanism at the interface of the Catch1 and Catch2 subdomains. This investigation generalizes and expands prior structural observations of MIDN-IRF4 to diverse substrates, demonstrating that β-strand complementation constitutes a universal recognition mechanism utilized by MIDN. A major conceptual advance of this study is the identification of a minimal and generalizable recognition principle centered on two highly conserved positions within the substrate motif. The authors show that a reciprocal phenylalanine-glycine interaction between the substrate and Catch2—forming an “F-G zipper”—constitutes the dominant energetic determinant for binding. Disruption of this zipper severely compromises protein stability and binding, explaining prior functional observations that single-point mutations in IRF4 or EGR1 abolish MIDN-mediated degradation. In contrast, flanking residues within the binding motif display remarkable tolerance to substitution. Through combined mutagenesis, thermostability analysis, AlphaFold3 modeling, and structural determination, the study demonstrates that these positions occupy large and plastic hydrophobic pockets in the Catch domain. This architectural flexibility provides a direct molecular explanation for how MIDN can accommodate a wide spectrum of substrates while preserving selectivity. From these data, the authors derive a concise consensus recognition motif (G/S-x-F/Y) embedded within an unstructured or loop region, offering a predictive framework for identifying additional MIDN targets. Importantly, this work bridges a critical gap between previous cryo-EM studies describing MIDN-proteasome engagement and earlier substrate-specific structural analyses. Whereas prior studies clarified how MIDN delivers captured substrates to the proteasome, the present study elucidates how MIDN initially selects and binds those substrates. Together, these findings unify MIDN biology across structural, biochemical and functional dimensions. Overall, this study represents a significant advance in the field of proteostasis by revealing how a ubiquitin-independent adaptor achieves broad substrate specificity through a simple yet robust structural principle. Beyond MIDN, the work provides a paradigm for short-linear-motif-based proteasomal targeting and has important implications for immune regulation, neurodegeneration and cancer biology.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2025171
Obesity-induced metabolic inflammation is a key driver of chronic kidney disease (CKD), with immune dysregulation, particularly among lymphocytes, contributing to early disease pathology. To explore the role of apolipoprotein A4 (Apoa4) in regulating immune cell metabolism and function, we establish high-fat diet-induced obese (DIO) models using wild-type and Apoa4-knockout (KO) mice. KO mice exhibit exacerbated insulin resistance and renal lipid accumulation. Single-cell RNA sequencing reveals that Apoa4 deletion remodeled the renal immune-metabolic landscape. This remodeling broadly compromises the immune functions of T, NK, and B cells, even as it expands the proportions of cytotoxic Gzma+ NK cells and Derl3+ plasma cells. Mechanistically, Apoa4 deletion aggravates metabolic dysregulation and oxidative stress and downregulates the expression levels of key effector genes, including Ifng and Il1b. Furthermore, the regulatory network activities of key transcription factors, such as Lef1 and Runx3 in Cd8+ T cells; Irf8, T-bet, and Eomes in NK cells; and Tcf4, Lmo2, and Xbp1 in B cells, are perturbed. CellChat analysis predicts disruptions in pro-inflammatory (IFN-II and IL-1), immunoregulatory (FASLG), and metabolic regulatory (ENHO and ANGPTL) signaling, alongside enhanced IL-2-mediated suppression. These findings are corroborated by flow cytometry, immunofluorescence staining, and qPCR. Our results establish Apoa4 as a crucial regulator of lymphocyte metabolic and immune homeostasis in the early stages of obesity-associated CKD.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2025048
As a classic tumor suppressor gene, p53 has been extensively studied since its discovery in the mid-1980s. Research findings have revealed that p53 protein expression is suppressed in various cancers [1]. For example, in cervical cancer, p53 predominantly exists in a wild-type form to maintain its biological function [2]. Nevertheless, its tumor-suppressive activity is significantly impaired because of rapid protein degradation, short half-life, and low levels. Post-translational modifications (PTMs) of p53, such as ubiquitination, acetylation, phosphorylation and methylation, are critical regulators of its stability, activity, conformation, localization, and interactions with cofactors [3]. Among these, ubiquitination and acetylation play central roles in controlling p53 protein stability and activity [4]. Therefore, targeting p53 PTMs to modulate its ubiquitination and acetylation levels represents an effective strategy to increase its stability and tumor-suppressive function, offering a promising avenue for cervical cancer drug development. In 99% of cervical cancers (high-risk human papillomavirus-positive), E3 ubiquitin ligase E6-associated protein (E6AP) mediates the ubiquitination degradation of p53 [5], whereas histone deacetylase 6 (HDAC6) deacetylates p53. In this study, we explored the possibility of combining the natural product withferin A (WA) with the HDAC6 inhibitor ricolinostat (RIC) to treat cervical cancer cells, with a focus on the ubiquitination and acetylation of p53 and the consequences for its stability. These results suggested that the combination of WA and RIC is more effective than either treatment alone in inhibiting the degradation and increasing the stability of p53, thereby synergistically slowing the onset and progression of cervical cancer.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2025047
Systemic lupus erythematosus (SLE) is an autoimmune disease characterized by a complex pathogenesis that was previously thought to involve primarily adaptive immunity. Emerging evidence underscores the role of neutrophils in shaping immune dysregulation and inducing organ damage in lupus. This study aims to investigate the dynamics of neutrophil senescence and its relationship with lupus, an area that remains poorly understood. Here, we identify a significantly elevated proportion of CXCR4hiCD62Llo senescence-like neutrophils in the peripheral blood of SLE patients compare to that in the healthy donors. Increased numbers of senescence-like neutrophils are positively correlated with SLE disease activity and autoantibody production in SLE patients. In addition, senescence-like neutrophils derived from SLE patients exhibit an impaired ability to suppress the proinflammatory activity of natural killer (NK) cells and CD4+ T cells. Further mechanistic exploration suggests that these senescence-like neutrophils might exert their immunosuppressive effects via reactive oxygen species (ROS) production under physiological conditions. Our results demonstrate that senescence-like neutrophils could serve as biomarkers for assessing the disease activity of SLE. The compromised immunosuppressive function of senescence-like neutrophils provides a new perspective on SLE pathophysiology and may pave the way for the development of novel therapies.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2025121
Carfilzomib (Cfz) is a second-generation proteasome inhibitor approved for the treatment of relapsed/refractory multiple myeloma (RRMM). Previous studies have shown that Cfz is associated with a higher incidence of severe adverse cardiac effects than bortezomib (Btz); however, the underlying mechanisms remain to be elucidated. The aim of this study is to identify key regulators of cardiotoxicity induced by Cfz and to investigate the mechanisms by which these factors exert their effects. We establish a mouse model of cardiac toxicity induced by Cfz and confirm the phenotype through cardiac functional analysis, morphology assessment, myocardial fibrosis, and apoptosis analysis. We subsequently perform RNA sequencing to identify differentially expressed genes (DEGs) and further validate their functions and mechanisms. We find that Cfz induces myocardial hypertrophy and myocardial injury, along with the suppression of SENP1 expression in mouse heart tissues and in vitro cultured neonatal rat cardiomyocytes. Suppression of SENP1 exacerbates Cfz-induced injury and remodeling in cardiomyocytes by directly binding to and deconjugating the SUMO1-mediated SUMOylation of the RNA helicase DDX17. This process leads to a reduction in K-48 ubiquitin-linked polyubiquitination and degradation of DDX17, resulting in increased expressions of anti-apoptotic genes and maintenance of mitochondrial homeostasis. Therefore, the overexpression of SENP1 using AAV vectors alleviates Cfz-induced cardiotoxicity in mice. In summary, our findings reveal a previously unknown role of the SENP1-DDX17 axis in protecting against cardiotoxicity induced by Cfz, providing a potential foundation for developing therapeutic strategies to mitigate cardiac side effects in the clinical management of MM patients.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2025193
Arsenic, a toxic metalloid, exists in organic or inorganic states within the Earth’s seawater, river water, soil, atmosphere, food sources, and diverse biological tissues [1]. It poses a threat to the health of hundreds of millions of people globally [2]. Arsenic exposure has toxic effects on the cardiovascular system of organisms, thus endangering human health [3]. Research has indicated that the harmful effect of arsenic exposure on the heart is associated with abnormal calcium handling in myocardial cells [4]. The cardiac ryanodine receptor type 2 (RyR2) is a primary channel involved in the surface of the endoplasmic reticulum in cardiac myocytes that regulates the release of Ca2+ during the systolic phase [5]. The integrity of its function is crucial for maintaining calcium homeostasis in cardiac myocytes. However, when myocardial tissue is damaged and undergoes pathological changes, the spatial structure of the RyR2 protein becomes unstable and becomes excessively activated, thereby triggering Ca2+ leakage [6]. Dantrolene (Dan), which serves as a stabilizer of RyR1, is frequently employed in clinical settings for the treatment of malignant hyperpyrexia and relieves spastic muscle tension [7]. Previous studies have demonstrated that dantrolene also has a stabilizing effect on RyR2 [8]. Research has shown that dantrolene can prevent calcium leakage in myocardial cells by stabilizing the tertiary structure of the RyR2 protein and thereby inhibiting the pathological hyperactivity of RyR2 [9]. Therefore, this study hypothesizes that dantrolene, by virtue of this stabilizing effect, can alleviate myocardial injury caused by arsenic exposure to some extent and plays a role in protecting cardiac function. For this purpose, we established an arsenic exposure model and a Dan intervention arsenic exposure model to verify the protective effect of Dan on the myocardial tissue and cardiac function of arsenic-exposed rats.
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•2025•DOI: 10.3724/abbs.2024179
The ubiquitin-proteasome pathway is a highly selective protein degradation pathway that is capable of efficiently degrading intracellular proteins and plays an important role in various life processes. Dysfunction of this pathway has been associated with numerous problems, including cancer and neurodegenerative diseases. Targeted protein degradation (TPD) technologies have emerged as promising tools for use in a number of different areas, including biological research and clinical interventions. Recently, a technology named Trim-Away was developed for the rapid degradation of proteins in mammalian cells. Briefly, an antibody is designed against a target protein, and the E3 ligase TRIM21 is used to recognize the Fc region of the antibody and subsequently mediate antibody-dependent protein degradation via the proteasome. To enhance the protein degradation efficiency of Trim-Away, three TRIM21-based constructs were designed: (1) deletion of the B-box domain of TRIM21, termed TRIM21 (ΔBB), (2) substitution of the RING domain of TRIM21 with the RING domain of MKRN1, termed TRIM21-RING, and (3) substitution of the RING domain of TRIM21 with the HECT domain of UBE3A, designated TRIM21-HECT. The antibody was designed as a human IgG Fc region-fused nanobody. To test the protein degradation efficiency of these TRIM21-based constructs, plasmids encoding the d2EGFP, an antibody against d2EGFP, and various Trim21-based constructs were co-transfected into HEK293T cells. The results revealed that TRIM21 (ΔBB) exhibited the most effective degradation performance, followed by TRIM21, whereas TRIM21-RING and TRIM21-HECT performed poorly. A dose-dependent assay confirmed that TRIM21 (ΔBB) showed the best degradation performance even at lower doses. Human papillomavirus (HPV) is a major contributor to the global burden of cancer, and high-risk subtypes are associated with approximately 90% of cervical cancers. Two viral oncoproteins, E6 and E7, play a role in carcinogenesis. Antibodies against E6 and E7 were designed and validated for their ability to degrade these proteins in HEK293T cells and in the cervical cancer cell line CaSki. The results showed that TRIM21 (ΔBB) exhibited the most effective degradation effect, and further investigation revealed that the TRIM21 (ΔBB) construct was able to degrade the E6 and E7 proteins.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2025169
AXL, a member of the TAM (Tyro3, AXL, and Mertk) subfamily of RTKs, is abundantly expressed in lung tissue and has been implicated in viral infections and lung injury. PROS1, one of the ligands known to activate AXL, functions as an immunomodulator in many diseases. However, the role of PROS1/AXL signaling in influenza A virus (IAV) infection and infection-induced lung injury is largely unknown. In this study, we find that the exogenous administration of PROS1 mitigates lung injury and protects mice from lethal infection by IAVs through the activation of AXL. PROS1 induces the phosphorylation of AXL, which in turn recruits Gab1 and p85, a regulatory subunit of PI3K, to form a complex that activates Gab1 and its downstream PI3K/AKT/mTOR in alveolar macrophages. Gab1 knockdown in vivo, or LY294002 (a PI3K inhibitor), abolishes the PROS1/AXL-induced protective activity against lethal influenza infection in mice. We also show that PROS1/AXL signaling induces M2 polarization of alveolar macrophages through Gab1 activation both in vitro and in vivo. Gab1 knockdown inhibits M2 macrophage accumulation in IAV-infected lungs and attenuates the protective effect of PROS1. These results indicate that PROS1/AXL signaling can activate Gab1 in macrophages and induce macrophage polarization to an anti-inflammatory M2 phenotype, thereby eliciting protective activity against lethal infection with IAVs. These data also highlight the PROS1/AXL signal as a novel therapeutic target for IAV infection.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2025101
Abnormal proliferation and migration of endothelial cells are key contributors to tumor angiogenesis. Recent studies have shown that the crucial role of E3 ubiquitin ligase neuronal precursor cell expression developmentally downregulated 4-like (NEDD4L) in tumorigenesis. However, the precise mechanisms by which NEDD4L functions in endothelial cells remain unclear. In this study, we investigate the mechanisms by which NEDD4L influences the function of human umbilical vein endothelial cells (HUVECs) and its effect on tumor angiogenesis. Our results show that NEDD4L overexpression in HUVECs suppresses both cell proliferation and migration. Additionally, we find that the autophagic activity in NEDD4L-overexpressing cells is increased. Proteomic profiling and ubiquitination assays reveal that NEDD4L interacts with eEF1A1, promoting K48-linked ubiquitination-mediated degradation of eEF1A1. This post-translational modification is a key step in the NEDD4L-mediated regulation of autophagy and cellular function. Moreover, we find that loss of endothelial NEDD4L significantly enhances tumor growth and promotes angiogenesis in vivo. Overall, NEDD4L plays a crucial role in inhibiting tumor angiogenesis by regulating eEF1A1 ubiquitination and degradation, providing new insights into the NEDD4L-eEF1A1 axis and its potential as a therapeutic target.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2024104
Carfilzomib (CFZ) is the second-generation proteasome inhibitor that is approved by Food and Drug Administration (FDA) of USA for the treatment of relapsed and refractory multiple myeloma. Although the preclinical and clinical efficacy of CFZ is obvious, the mechanism by which CFZ leads to cell death has not been fully elucidated. Since CFZ primarily functions as a proteasome inhibitor, profiling CFZ-induced changes in protein turnover at the systematic level is sufficient and necessary. In this study, we characterize the effects of CFZ on the stability of 15,000 human proteins using Protein Turnover Assay (ProTA). CFZ affects fundamental cellular glycolysis, nitric oxide production and proteasome subunit homeostasis in multiple myeloma cells. In addition, LY294002 or KU-0063794 has synergistic effects with CFZ in multiple myeloma treatment. A profound understanding of how cells respond to chemotherapeutic agents provides insights into the basic mechanism of drug function and the rationale for CFZ combination therapy.
Acta Biochimica et Biophysica Sinica•2024•DOI: 10.3724/abbs.2024087
The aberrant proliferation and migration of vascular smooth muscle cells (VSMCs) contribute to the development of neointima formation in vascular restenosis. This study aims to explore the function of the long noncoding RNA H19 in neointima formation. A mouse carotid ligation model was established, and human vascular smooth muscle cells (VSMCs) were used as a cell model. lncRNA H19 overexpression promoted VSMC proliferation and migration. Moreover, miR-125a-3p potentially bound to lncRNA H19, and Fms-like tyrosine kinase-1 (FLT1) might be a direct target of miR-125a-3p in VSMCs. Upregulation of miR-125a-3p alleviated lncRNA H19-enhanced VSMC proliferation and migration. Furthermore, rescue experiments showed that enhanced expression of miR-125a-3p attenuated lncRNA H19-induced FLT1 expression in VSMCs. In addition, the overexpression of lncRNA H19 significantly exacerbated neointima formation in a mouse carotid ligation model. In summary, lncRNA H19 stimulates VSMC proliferation and migration by acting as a competing endogenous RNA (ceRNA) of miR-125a-3p. lncRNA H19 may be a therapeutic target for restenosis.
Acta Biochimica et Biophysica Sinica•2024•DOI: 10.3724/abbs.2024076
Rheumatoid arthritis (RA) is an idiopathic and chronic autoimmune disease for which there are currently no effective treatments. Oxypeucedanin hydrate (OXH) is a natural coumarin known for its potent anti-inflammatory properties. However, further investigations are needed to determine its therapeutic efficacy in treating RA. In this study, we evaluate the anti-inflammatory activity of OXH by treating LPS-induced RAW264.7 macrophages. Our results show that OXH treatment reverses the changes in iNOS, COX-2, IL-1β, IL-6, and TNF-α levels. Additionally, OXH reduces ROS production. Further analysis reveals that OXH suppresses the activation of the NF-κB/MAPK pathway. CETSA results show that OXH competes with LPS for binding to the TLR4/MD2 complex. MST experiments demonstrate the specific affinity of OXH for the TLR4/MD2 complex, with a Kd value of 33.7 μM. Molecular docking analysis suggests that OXH binds to the pocket of the TLR4/MD2 complex and interacts with specific amino acids, such as GLY-343, LYS-388, and PHE-345. Molecular dynamics simulations further confirm this conclusion. Finally, we investigate the potential of OXH in treating RA using a collagen-induced arthritis (CIA) model in rats. OXH effectively ameliorates the symptoms of CIA, including improving body weight, reducing swelling and redness, increasing talus volume, and decreasing bone erosion. OXH also decreases the mRNA levels of pro-inflammatory factors in synovial tissue. Transcriptome enrichment analysis and western blot analysis confirm that OXH suppresses the NF-κB/MAPK pathway, which is consistent with our in vitro findings.
Chinese Journal of Pathophysiology•2025•DOI: 10.3969/j.issn.1000-4718.2025.09.002
AIM: To investigate whether melatonin can ameliorate acute myocardial infarction (AMI) by inhibiting ferroptosis. METHODS: H9C2 cells were cultured in AnaeroPack system with low sugar and serum-free medium for 10 h to construct a cell model of AMI. Then cells were treated with melatonin and ferroptosis inducer erastin. The cell activity, reactive oxygen species (ROS), lipid peroxidation, mitochondrial membrane potential (MMP), and ferroptosis related protein expression were detected. A rat model of AMI induced by isoprenaline (ISO) injection was established to evaluate the effects of melatonin, in which the myocardial infarction size, cardiac injury, pathological changes, oxidative stress, iron ion and ferroptosis related protein expression were examined. RESULTS: Melatonin decreased the oxidative stress, lipid peroxidation and expression of ferroptosis protein in cardiomyocytes induced by hypoxia, but these effects could be impeded by the ferroptosis inducer erastin. Furthermore, in vivo experiments, we also found that melatonin improved the myocardial infarction size, cardiac injury, pathological changes, oxidative stress, and alleviated iron ion accumulation and ferroptosis. CONCLUSION: The cardioprotective effects of melatonin in AMI are associated with the inhibition of ferroptosis.
Chinese Traditional and Herbal Drugs•2026•DOI: 10.7501/j.issn.0253-2670.2026.16.20261615
Ulcerative colitis (UC) remains a clinical challenge due to inadequate mucosal healing and high relapse rates. This study investigates the therapeutic efficacy of schisandrin B (Sch B) in a 2,4,6-trinitrobenzenesulfonic acid (TNBS)-induced rat model of UC, focusing on the interplay between autophagy and pyroptosis. SD rats were randomized into control, model, mesalazine (100 mg/kg), and Sch B low-, medium-, and high-dose (10, 20, 50 mg/kg) groups (n=10 per group). After 14 days of treatment, disease activity index (DAI), colon length, and colon mucosa damage index (CMDI) were assessed. Histopathology, serum cytokine levels (TNF-α, IL-6, IL-1β), and protein expression of autophagy markers (Beclin-1, LC3B, ATG16L1, p62) and pyroptosis pathway components (NLRP3, Caspase-1, GSDMD) were evaluated. Sch B significantly ameliorated weight loss, hematochezia, and colon shortening (P<0.05, 0.01), reduced DAI and CMDI scores, and attenuated mucosal edema, ulceration, and inflammatory infiltration. Serum IL-6, TNF-α, and IL-1β levels were markedly decreased (P<0.05, 0.01). Sch B upregulated Beclin-1 and increased LC3-II/I ratio (P<0.01), while downregulating ATG16L1, p62, NLRP3, Caspase-1, and GSDMD (P<0.05, 0.01). These findings indicate that Sch B restores autophagic flux homeostasis, thereby suppressing NLRP3/Caspase-1/GSDMD-mediated pyroptosis and reducing pro-inflammatory cytokine release. The normalization of autophagic flux is a critical upstream mechanism for Sch B's inhibition of colonic epithelial pyroptosis, offering a multi-target therapeutic strategy for UC.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2026006
Protein homeostasis is fundamental to cellular decisions, and its dysregulation drives numerous pathologies. The ubiquitin-independent proteasomal degradation pathway mediated by midnolin (MIDN) has emerged as a distinct mechanism for regulating nuclear protein turnover. In the current study, Zhong et al. provide a comprehensive structural and biochemical analysis of the MIDN Catch domain bound to IRF4-derived peptides, substantially advancing our understanding of MIDN substrate recognition. By solving multiple crystal structures of the Catch-IRF4 complex, the authors demonstrate that MIDN recognizes substrates through a conserved β-strand insertion mechanism at the interface of the Catch1 and Catch2 subdomains. A major conceptual advance is the identification of a minimal and generalizable recognition principle centered on two highly conserved positions within the substrate motif, forming an 'F-G zipper' that constitutes the dominant energetic determinant for binding. In contrast, flanking residues display remarkable tolerance to substitution, occupying large and plastic hydrophobic pockets in the Catch domain. This architectural flexibility provides a direct molecular explanation for how MIDN can accommodate a wide spectrum of substrates while preserving selectivity. The authors derive a concise consensus recognition motif (G/S-x-F/Y) embedded within an unstructured or loop region, offering a predictive framework for identifying additional MIDN targets. Importantly, this work bridges a critical gap between previous cryo-EM studies describing MIDN-proteasome engagement and earlier substrate-specific structural analyses. Overall, this study represents a significant advance in the field of proteostasis by revealing how a ubiquitin-independent adaptor achieves broad substrate specificity through a simple yet robust structural principle, with important implications for immune regulation, neurodegeneration, and cancer biology.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21207
BACKGROUND: Hypoxia preconditioning can increase extracellular vesicles, growth factors, anti-inflammatory and immunosuppressive factors in the parasecretory secretions of mesenchymal stem cells. Exosomes derived from human amniotic mesenchymal stem cells preconditioned with hypoxia are expected to play a better therapeutic role in tissue damage repair. OBJECTIVE: To observe the repair effect of exosomes from human amniotic mesenchymal stem cells preconditioned with hypoxia on radiation-induced submandibular gland epithelial cell damage. METHODS: The passage 3 human amniotic mesenchymal stem cells were divided into two groups: hypoxia and normoxia. They were pretreated with hypoxia (1% O2 by volume) and normoxia (20% O2 by volume) for 48 hours, respectively. Then, normoxic and hypoxic human amniotic mesenchymal stem cell exosomes were extracted from the culture supernatant of human amniotic mesenchymal stem cells by modified ultrahigh-speed centrifugation. The submandibular gland epithelial cells of SD newborn rats were divided into blank control group, radiation control group, normoxia-treated group, and hypoxia-treated group. The submandibular gland epithelial cells were treated with 5 Gy radiation to construct a radiation-induced injury model. The normoxia-treated and hypoxia-treated groups were co-cultured with normoxic or hypoxic exosomes for 3 days. CCK-8 assay was used to detect the proliferation activity of rat submandibular gland epithelial cells; ELISA was used to measure the content of α-amylase in the supernatant; RT-qPCR was used to detect the expression of aquaporin 5 mRNA. RESULTS AND CONCLUSION: The cell proliferation activity in the radiation control group was significantly lower than that in the blank control group (P < 0.05). The cell proliferation activity in the normoxia-treated and hypoxia-treated groups on days 2 and 3 was significantly higher than that in the radiation control group (P < 0.05). The cell proliferation activity in the hypoxia-treated group on day 1 was higher than that in the normoxia-treated group, and showed an increasing trend on days 2 and 3, but there was no statistical difference compared with the normoxia-treated group (P > 0.05). The α-amylase content in the radiation control group was significantly lower than that in the blank control group (P < 0.05). The α-amylase content in the normoxia-treated and hypoxia-treated groups was significantly higher than that in the radiation control group (P < 0.05). Compared with the normoxia-treated group, the α-amylase content in the hypoxia-treated group showed an increasing trend, but there was no statistical difference (P > 0.05). The expression of aquaporin 5 mRNA in the radiation control group was significantly lower than that in the blank control group (P < 0.05). The expression of aquaporin 5 mRNA in the normoxia-treated and hypoxia-treated groups was significantly higher than that in the radiation control group (P < 0.05). The expression of aquaporin 5 mRNA in the hypoxia-treated group was slightly higher than that in the normoxia-treated group, but there was no statistical difference (P > 0.05). The results indicate that both normoxic and hypoxic preconditioned human amniotic mesenchymal stem cell exosomes have a repair effect on radiation-induced submandibular gland epithelial cell damage, but hypoxic exosomes have no obvious advantage.
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: 10.12307/2026.21376
OBJECTIVE: This study aimed to systematically compare the efficacy and safety of various neuromodulation techniques for upper limb motor function recovery after stroke, and to rank the relative advantages of different interventions through a network meta-analysis, thereby providing evidence-based guidance for clinical rehabilitation. METHODS: A comprehensive literature search was conducted in CNKI, WanFang, VIP, CBM, PubMed, EMbase, Web of Science, and Cochrane Library from inception to August 2025. Randomized controlled trials investigating different neuromodulation techniques for post-stroke upper limb motor impairment were included. Control group received sham stimulation or conventional rehabilitation, while trial group received additional neuromodulation therapies. The methodological quality of included studies was assessed using the Cochrane Risk of Bias Tool. Network meta-analyses were performed using Stata 16.0 and RevMan 5.4 software. RESULTS: A total of 51 randomized controlled trials were included, covering 12 neuromodulation stimulation modalities. Network meta-analysis results showed that compared with conventional treatment, high-frequency repetitive transcranial magnetic stimulation (MD=11.50, 95%CI: 6.83-16.16, P < 0.05) was most effective in improving basic upper limb motor function recovery; continuous theta burst stimulation (MD=12.10, 95%CI: 44.99-19.21, P < 0.05; MD=9.60, 95%CI: 1.32-17.88, P < 0.05) was most effective in improving the practicality and dexterity of upper limb function; and cathodal transcranial direct current stimulation (MD=15.40, 95%CI: 0.03-30.77, P < 0.05; MD=-0.83, 95%CI: -1.64 to -0.03, P < 0.05) was most effective in improving daily living activity limitations or with obvious spasticity. CONCLUSION: When the goal is to promote basic upper limb motor function recovery, high-frequency repetitive transcranial magnetic stimulation is most effective; for improving the practicality and dexterity of upper limb function, continuous theta burst stimulation is most effective; and for patients with impaired daily living ability accompanied by obvious spasticity, cathodal transcranial direct current stimulation is most effective.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21355
BACKGROUND: In recent years, multiple epidemiological studies have suggested a potential pathological link between sarcopenia and cognitive impairment. However, due to methodological limitations in traditional observational studies and difficulties in controlling confounding factors, their genetic-level causal relationship has not yet been fully elucidated. OBJECTIVE: To systematically analyze the causal relationship and underlying pathogenesis between sarcopenia and cognitive impairment in European populations using Mendelian randomization methods. METHODS: This study utilized genome-wide association study (GWAS) summary data for sarcopenia-related phenotypes (whole-body fat-free mass, hand grip strength, and walking speed) from the UK Biobank, and cognitive function GWAS summary data from the IEU database. After rigorous threshold filtering and linkage disequilibrium clumping, bidirectional Mendelian randomization analyses were performed. Forward analysis used sarcopenia-related traits as exposures and cognitive function as the outcome; reverse analysis swapped the direction. Inverse variance weighting was the primary analysis method, supplemented by weighted median, MR-Egger regression, and robust adjusted profile scoring. Heterogeneity and sensitivity analyses were conducted to ensure robustness. RESULTS AND CONCLUSION: Forward MR-IVW analysis showed that whole-body fat-free mass (OR=1.091, 95%CI: 1.001-1.188, P=0.045), left hand grip strength (OR=1.283, 95%CI: 1.077-1.527, P=0.005), right hand grip strength (OR=1.220, 95%CI: 1.022-1.456, P=0.027), and walking speed (OR=3.069, 95%CI: 1.997-4.717, P<0.001) were significantly positively associated with cognitive function. Reverse analysis showed that cognitive function had a significant positive causal effect only on walking speed (OR=1.023, 95%CI: 1.004-1.043, P=0.014), but not on fat-free mass or grip strength. Sensitivity analyses indicated some heterogeneity but no horizontal pleiotropy. The findings suggest a causal relationship between sarcopenia and cognitive impairment, indicating that sarcopenia may serve as a predictor for cognitive impairment, providing a theoretical basis for early clinical screening. This study, based on international public databases, offers new evidence for the association between sarcopenia and cognitive impairment in Chinese populations and has important reference value for early screening and prevention of both diseases.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21314
BACKGROUND: Preliminary studies have demonstrated that Shaoyang Shenggu Fang can alleviate joint cartilage degeneration and promote cartilage repair, but its specific mechanism for alleviating knee osteoarthritis symptoms remains unclear. The Wnt/β-catenin pathway and oxidative stress play crucial roles in maintaining articular cartilage homeostasis. OBJECTIVE: To investigate the molecular mechanisms by which Shaoyang Shenggu Fang regulates the Wnt/β-catenin pathway to inhibit oxidative stress in cartilage and thereby delay cartilage aging in a rat model of knee osteoarthritis. METHODS: Thirty-two Sprague-Dawley rats were randomly divided into four groups: a blank control group, a model group, a Western medicine group, and a Chinese medicine group. Animal models of knee osteoarthritis were established in all groups except for the blank control group by transecting the anterior cruciate ligament and resecting the anterior horn of the medial meniscus. After 28 days of modeling, the Chinese medicine group was administered concentrated Shaoyang Shenggu Fang at a dose of 16 g/(kg·d) by gavage, the Western medicine group received glucosamine hydrochloride solution at 4 mL/d, and the blank and model groups received the same volume of normal saline. After 4 weeks, hematoxylin-eosin staining and Safranin O-fast green staining were used to observe the degree of cartilage damage and degeneration. ELISA was used to detect serum levels of inflammatory factors and oxidative stress indicators. Western blot was used to detect the expression of p21Cip1, p16INK4a, and Wnt signaling pathway-related proteins in knee cartilage. RESULTS AND CONCLUSION: Compared with the model group, the Western medicine and Chinese medicine groups showed significant improvement in cartilage defects, thinning of the cartilage layer, and decreased density, with significantly lower Mankin scores (P < 0.05). Compared with the model group, serum levels of interleukin-1β, tumor necrosis factor-α, and interleukin-6 were significantly decreased in the Western medicine and Chinese medicine groups (P < 0.05), while superoxide dismutase and glutathione peroxidase levels were increased and malondialdehyde concentration was decreased (all P < 0.05). In the Chinese medicine group, the expression levels of p21Cip1, p16INK4a, and Wnt5a proteins were significantly decreased (P < 0.05 and P < 0.01), β-catenin and C-Myc protein expression levels were decreased (P < 0.05), and glycogen synthase kinase-3β protein expression was significantly increased (P < 0.05). These results suggest that Shaoyang Shenggu Fang can significantly reduce inflammation and alleviate cartilage aging in rats with knee osteoarthritis, and the potential mechanism may be through regulation of the Wnt/β-catenin pathway to inhibit cartilage oxidative stress.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21444
BACKGROUND: Icariin has the dual activity of promoting bone formation and inhibiting bone resorption, but its clinical application is plagued by low bioavailability, difficulty in controlling dosage, and a high risk of adverse reactions. OBJECTIVE: To prepare a three-dimensional scaffold containing icariin sustained-release microspheres and characterize their osteogenic activity in vitro. METHODS: A silk fibroin/chitosan/nanohydroxyapatite three-dimensional scaffold (SF/CS/nHA scaffold), icariin sustained-release microspheres, and a silk fibroin/chitosan/nanohydroxyapatite three-dimensional scaffold loaded with icariin sustained-release microspheres (SF/CS/nHA-ICA scaffold) were prepared. The drug loading efficiency, encapsulation efficiency, and in vitro drug release of the icariin sustained-release microspheres were characterized. The pore size, porosity, water absorption expansion rate, and hot water dissolution rate of the two scaffolds were measured. Rabbit bone marrow mesenchymal stem cells (or human rheumatoid arthritis fibroblast-like synoviocytes) were seeded on SF/CS/nHA and SF/CS/nHA-ICA scaffolds, with cells cultured alone as controls. Cell adhesion was observed by scanning electron microscopy. Cell proliferation and viability were assessed by CCK-8 assay, live/dead staining, and F-actin staining. The mRNA and protein expression of Runx-2, osteocalcin, and type I collagen in bone marrow mesenchymal stem cells were detected by RT-qPCR and western blot. RESULTS AND CONCLUSION: (1) The drug loading efficiency and encapsulation efficiency of icariin sustained-release microspheres were (29.38±0.04)% and (52.01±0.09)%, respectively, and the microspheres could sustainably release icariin for more than 90 days in vitro. (2) Scanning electron microscopy showed a honeycomb-like porous structure with interconnected pores in both scaffolds. There were no significant differences in pore size, porosity, water absorption expansion rate, or total hot water dissolution rate between the two groups (P > 0.05). (3) Scanning electron microscopy showed that both cell types adhered tightly to the scaffold surface and pores, with more extended pseudopodia on the SF/CS/nHA-ICA scaffold. CCK-8 assay, live/dead staining, and F-actin staining showed that compared with the control and SF/CS/nHA groups, the SF/CS/nHA-ICA scaffold promoted the proliferation and viability of rabbit bone marrow mesenchymal stem cells, while inhibiting the proliferation and viability of human rheumatoid arthritis fibroblast-like synoviocytes. (4) RT-qPCR and western blot showed that compared with the control and SF/CS/nHA groups, the mRNA and protein expression of Runx-2, osteocalcin, and type I collagen were increased in the SF/CS/nHA-ICA group (P < 0.05). (5) These results indicate that the icariin sustained-release microsphere three-dimensional scaffold has good cytocompatibility, and in vitro osteogenic and anti-inflammatory effects.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21522
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: 10.12307/2026.21594
BACKGROUND: In recent years, hydrogel scaffolds, as important carriers in the field of tissue engineering, have made significant progress in their application in urethral repair and reconstruction. OBJECTIVE: To systematically review the design strategies of urethral tissue engineering scaffolds, the functional modification of hydrogel scaffolds, and their application progress in urethral repair, and to explore the future development directions and clinical transformation challenges of hydrogel scaffolds. METHODS: The CNKI and PubMed databases were searched using Chinese and English keywords "hydrogel, tissue engineering scaffold, urethral tissue engineering, urethral repair, urethral reconstruction." Based on the inclusion criteria, 69 articles were finally selected for inductive analysis. RESULTS AND CONCLUSION: In urethral tissue engineering, urethral scaffolds should not only possess basic characteristics such as biocompatibility but also exhibit unique properties that adapt to the structure of the urethra and the urinary environment. Hydrogel scaffolds can be classified into natural hydrogel scaffolds (commonly protein-based, polysaccharide-based, DNA-based, and extracellular matrix-based) and synthetic polymer hydrogel scaffolds (commonly polyvinyl alcohol, polyacrylic acid, acrylate-based). Crosslinking methods mainly include physical and chemical crosslinking, and preparation methods include in situ gelation, freeze-drying, electrospinning, and 3D bioprinting. To avoid the drawbacks of single materials, composite hydrogel scaffolds are often used in urethral tissue engineering, such as composite scaffolds adapted to the urethral microenvironment, composite scaffolds utilizing decellularized extracellular matrix, multilayer composite scaffolds loaded with stem cells, and non-stem cell-based hydrogel scaffolds. With the development of tissue engineering and regenerative medicine, future hydrogel scaffolds should be designed with intelligent, engineered, and cell-free strategies, while considering current clinical translation barriers and proposing comprehensive solutions.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2026045
Colorectal cancer (CRC) remains a leading cause of cancer-related mortality, necessitating the discovery of novel therapeutic agents. Here, we report a natural small molecule, 2-dihydroailanthone (2-DAIL), as a promising candidate for CRC treatment. First, our results demonstrate that 2-DAIL exhibits significant anti-CRC activity in vitro and in vivo. Then, we find that 2-DAIL directly binds to integrin alpha-3 (ITGA3) revealed by stable isotope labeling by amino acids in cell culture coupled with thermal proteome profiling (SILAC-TPP). Additionally, the RNA sequencing data obtained from CRC cells and tumor tissues suggest that 2-DAIL blocks the PI3K/AKT signaling pathway mediated by ITGA3 inhibition. Collectively, 2-DAIL exerts its anti-CRC effects, at least partially, by binding to and inhibiting the function of ITGA3, thereby blocking the activation of the PI3K/AKT signaling pathway, which leads to CRC cell growth inhibition. Our study provides a promising drug candidate for the treatment of CRC and suggests the potential of 2-DAIL in treating other diseases linked to ITGA3 dysfunction.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2025232
Midnolin (MIDN) is a newly recognized master regulator that drives ubiquitin-independent proteasomal degradation, yet the mechanisms governing its own turnover remain enigmatic. Here, we demonstrate that MIDN is ubiquitinated and identify RNF126 as the cognate E3 ligase. RNF126 physically associates with MIDN and catalyzes its ubiquitination, and mass spectrometry mapping reveals that this process occurs primarily at non-canonical cysteine, serine, and threonine residues (C230, C236, S237, T239, and S241) rather than at lysine residues. This non-classical ubiquitination targets MIDN for 26S-proteasomal degradation. In vivo dissection of the RNF126-MIDN axis shows that it governs EGR1 abundance and, consequently, the tumor-suppressor proteins PTEN and p53, thereby restraining the progression of testicular germ-cell tumors (TGCTs). Our findings reveal an unappreciated layer of MIDN regulation and identify the RNF126-MIDN ubiquitination cascade as a potential therapeutic vulnerability in TGCTs and related malignancies.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2026033
Metabolic dysfunction-associated steatohepatitis (MASH) has become a global epidemic, and effective therapeutic strategies are urgently needed. Lonidamine (LND) has been reported to possess anti-inflammatory effects; however, few studies have investigated whether LND exerts a therapeutic effect on MASH. Therefore, in this study, we aim to explore the effects of LND on inflammatory responses and abnormal lipid metabolism in MASH mice. A mouse MASH model is established by feeding C57BL/6 mice a high-fat, high-cholesterol (CL) diet. The results show that LND attenuates CL-induced increases in body weight, serum glucose and lipid levels, inflammatory responses, and hepatocellular steatosis. In addition, the mitogen-activated protein kinase (MAPK) signaling pathway is inhibited, and the expression level of sterol regulatory element-binding protein 1 (SREBP1) protein is significantly reduced. Meanwhile, in vitro models of cellular inflammation and lipid metabolism are simulated, and molecular docking and biolayer interferometry (BLI) analysis are used to verify that LND and SREBP1 have a direct interaction and that LND promotes the degradation of SREBP1. Furthermore, specific knockdown of Srebp1 in AML12 cells is performed to further verify the effect of LND on MASH. The results confirm that LND exerts anti-inflammatory effects in MASH by inhibiting the activity of the MAPK signaling pathway and improves abnormal lipid metabolism through its interaction with SREBP1. Overall, LND holds promise as a potential therapeutic agent for the treatment of MASH.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2026099
Melanoma is an aggressive skin cancer with poor prognosis in advanced stages due to its metastatic potential. Although treatment modalities such as surgical resection, targeted therapy, immunotherapy, radiotherapy, and chemotherapy have conferred long-term survival benefits to certain patients, not all individuals respond favorably, underscoring the urgent need to develop predictive biomarkers. Ribosomal proteins have been implicated in tumor progression through extraribosomal functions. Ribosomal protein L8 (RPL8) is a unique ribosomal protein that warrants further exploration. Here, we investigated the role of RPL8 in melanoma progression. Using clinical data from the GDC and GEO databases, we found that high RPL8 expression is associated with poor prognosis in melanoma patients. Immunohistochemical analysis confirmed higher RPL8 expression in melanoma tissues compared to normal tissues. To elucidate the functional impact, we established an A875 melanoma cell line with stable RPL8 knockdown (shRPL8) using lentiviral vectors. RT-qPCR and western blot analysis confirmed efficient knockdown. CCK-8 assays showed that RPL8 knockdown significantly inhibited cell proliferation. Flow cytometry analysis revealed that RPL8 knockdown led to an increase in the proportion of cells in the G2/M phase and a decrease in the S phase, indicating cell cycle arrest. Furthermore, transwell assays demonstrated that RPL8 knockdown reduced cell invasion and migration. These findings suggest that RPL8 promotes melanoma progression by regulating the cell cycle and metastasis, and may serve as a potential therapeutic target and prognostic biomarker for melanoma.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2025047
Systemic lupus erythematosus (SLE) is a prototypic autoimmune disease with complex pathogenesis historically attributed to adaptive immunity. Emerging data implicate neutrophils in immune dysregulation and organ damage. This study investigates neutrophil senescence dynamics in SLE. We identified a significantly elevated proportion of CXCR4hiCD62Llo senescence-like neutrophils in peripheral blood of SLE patients versus healthy donors. Increased senescence-like neutrophil numbers positively correlated with SLE disease activity and autoantibody production. Functionally, senescence-like neutrophils from SLE patients exhibited impaired suppression of proinflammatory activity in natural killer (NK) cells and CD4+ T cells. Mechanistically, these cells may exert immunosuppressive effects via reactive oxygen species (ROS) production under physiological conditions. Our results position senescence-like neutrophils as candidate biomarkers for SLE disease activity. The compromised immunosuppressive function of these cells offers a new perspective on SLE pathophysiology and may inform development of novel therapies. Limitations include small sample size and heterogeneous treatment backgrounds, necessitating further validation. Future studies will address NET release and potential subset markers.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2025048
Cervical cancer remains a leading cause of cancer-related mortality among women, with high-risk HPV-positive cases constituting 99% of instances. Despite wild-type p53 expression, its tumor-suppressive function is crippled by E6AP-mediated ubiquitination and HDAC6-driven deacetylation, resulting in rapid degradation and low steady-state levels. This study evaluates a combinatorial strategy employing the natural product withaferin A (WA) and the HDAC6 inhibitor ricolinostat (RIC) to simultaneously target p53 ubiquitination and acetylation in HeLa, SiHa, and Caski cervical cancer cells. Dose-response CCK-8 assays established that both agents inhibit proliferation in a dose-dependent manner. Combination treatment significantly reduced cell viability compared to monotherapies, with CompuSyn analysis yielding a combination index (CI) below 1, confirming synergy. Colony formation assays further demonstrated a marked decrease in clonogenic survival. Mechanistically, WA disrupted the p53-E6AP interaction, reducing p53 ubiquitination, while RIC inhibited HDAC6-mediated deacetylation, increasing p53 acetylation. The dual treatment stabilized p53, as evidenced by extended half-life in cycloheximide chase assays. These findings suggest that concurrent modulation of p53 post-translational modifications via WA and RIC offers a potent therapeutic avenue for cervical cancer, meriting further preclinical development.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2025171
Obesity-induced metabolic inflammation drives chronic kidney disease (CKD), with lymphocyte dysregulation contributing to early pathology. We established high-fat diet-induced obese (DIO) models in wild-type and Apoa4-knockout (KO) mice to investigate apolipoprotein A4 (Apoa4) in immune-metabolic regulation. KO mice exhibited exacerbated insulin resistance and renal lipid accumulation. Single-cell RNA sequencing (scRNA-seq) of renal immune cells revealed that Apoa4 deletion remodeled the immune-metabolic landscape, compromising T, NK, and B cell functions while expanding cytotoxic Gzma+ NK cells and Derl3+ plasma cells. Mechanistically, Apoa4 deletion aggravated metabolic dysregulation and oxidative stress, downregulating effector genes including Ifng and Il1b. Transcription factor regulatory networks were perturbed: Lef1 and Runx3 in Cd8+ T cells; Irf8, T-bet, and Eomes in NK cells; and Tcf4, Lmo2, and Xbp1 in B cells. CellChat predicted disrupted pro-inflammatory (IFN-II, IL-1), immunoregulatory (FASLG), and metabolic (ENHO, ANGPTL) signaling, with enhanced IL-2-mediated suppression. Flow cytometry, immunofluorescence, and qPCR validated these findings. Sequencing depth averaged 278,276 reads/cell (WT) and 197,768 reads/cell (KO), ensuring robust detection of low-abundance transcripts despite modest cell capture. Apoa4 is a critical regulator of lymphocyte metabolic and immune homeostasis in early obesity-associated CKD.
Chinese Journal of New Drugs•2025•DOI: cast_zgxyzz_1236731781232251260
Background: Iron deficiency anemia (IDA) is a global health concern, and intravenous iron preparations are increasingly used. Ferric derisomaltose (FDI) is a newer formulation with potential advantages. This systematic review and meta-analysis aimed to evaluate the efficacy and safety of FDI compared with other iron therapies or placebo in treating IDA. Methods: We searched PubMed, Embase, Cochrane Library, and CNKI up to October 2023 for randomized controlled trials (RCTs) comparing FDI with active comparators or placebo in patients with IDA. The primary outcomes were change in hemoglobin (Hb) from baseline, and safety outcomes included adverse events (AEs), serious adverse events (SAEs), and hypersensitivity reactions. Data were pooled using random-effects models. Results: A total of 15 RCTs involving 3,452 patients were included. FDI significantly increased Hb levels compared with placebo (mean difference [MD] 1.2 g/dL, 95% CI 0.8-1.6) and was non-inferior to other intravenous iron formulations (MD 0.1 g/dL, 95% CI -0.2 to 0.4). FDI was associated with fewer hypersensitivity reactions compared with ferric carboxymaltose (risk ratio [RR] 0.3, 95% CI 0.1-0.9). The incidence of AEs was similar between FDI and other iron preparations. Subgroup analyses showed consistent results across different etiologies of IDA. Conclusion: Ferric derisomaltose is effective and safe for treating IDA, with a lower risk of hypersensitivity reactions compared with some other intravenous iron formulations. These findings support its use in clinical practice.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2025201
Colorectal cancer (CRC) remains the third most common malignancy and second leading cause of cancer mortality worldwide, with 608,000 new cases and 202,000 deaths annually in China. The Warburg effect, characterized by elevated glucose consumption and lactate production under normoxia, sustains CRC proliferation, migration, chemoresistance, and stemness. Prior work established that SUN5, a nuclear membrane protein, promotes CRC proliferation and migration, but its role in glycolysis remained undefined. Here, SUN5 overexpression increased glucose uptake and lactate production, whereas SUN5 knockdown produced reciprocal reductions. Mechanistically, SUN5 activates NF-κB signaling, an effect abolished by the IKK inhibitor BAY11-7082. SUN5 interacts with TRIM28 to enhance IκBα ubiquitination, driving nuclear translocation of phosphorylated P65 and transcriptional upregulation of GLUT1 and LDHA. Xenograft transplantation confirmed that SUN5 knockdown suppresses glycolysis and tumorigenesis in vivo. These findings position the SUN5–TRIM28–IκBα–NF-κB axis as a tractable target for CRC diagnosis and therapy.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2025062
p62/SQSTM1 is the archetypal selective autophagy receptor, bridging ubiquitinated cargo to LC3 on phagophores. Its abundance is a critical determinant of autophagic flux, yet the E3 ligase governing its turnover remained incompletely defined. Using Flag-p62 Co-IP coupled to mass spectrometry in HEK293T cells, we identified the E3 ligases TRIM25 and ITCH as highest-confidence interactors. Endogenous and ectopic p62 formed complexes with both ligases; GST pull-down confirmed direct binding, and mCherry-TRIM25 co-localized with GFP-p62 in HeLa cytoplasm. In vitro ubiquitination demonstrated that TRIM25, but not ITCH, efficiently ubiquitinates p62. A reconstituted E. coli system mapped fifteen lysine residues, with K7 and K189 validated as the dominant TRIM25-mediated ubiquitination sites. Functionally, TRIM25 destabilized wild-type p62 but not the K7/189R mutant; TRIM25 knockdown stabilized p62 in HeLa and Caski cells. Degradation proceeded primarily via the lysosomal pathway, as bafilomycin (20 nM) but not bortezomib (1 μM) blocked p62 loss. TRIM25 knockdown enhanced GFP-LC3 puncta formation (P < 0.01) and elevated autophagic markers, whereas TRIM25 overexpression suppressed p62-mediated GFP-LC3 puncta (P < 0.05) and autophagy. These data establish TRIM25 as the principal E3 ligase targeting p62 for lysosomal degradation, defining a negative feedback node in selective autophagy with therapeutic implications for cancers and neurodegenerative disorders characterized by p62 accumulation.