Genomics, Proteomics & Bioinformatics•2024•DOI: 10.1093/gpbjnl/qzad002
The order Acipenseriformes, which includes sturgeons and paddlefishes, represents “living fossils” with complex genomes that are good models for understanding whole-genome duplication (WGD) and ploidy evolution in fishes. Here, we sequenced and assembled the first high-quality chromosome-level genome for the complex octoploid Acipenser sinensis (Chinese sturgeon), a critically endangered species that also represents a poorly understood ploidy group in Acipenseriformes. Our results show that A. sinensis is a complex autooctoploid species containing four kinds of octovalents (8n), a hexavalent (6n), two tetravalents (4n), and a divalent (2n). An analysis taking into account delayed rediploidization reveals that the octoploid genome composition of Chinese sturgeon results from two rounds of homologous WGDs, and further provides insights into the timing of its ploidy evolution. This study provides the first octoploid genome resource of Acipenseriformes for understanding ploidy compositions and evolutionary trajectories of polyploid fishes.
Genomics, Proteomics & Bioinformatics•2024•DOI: 10.1093/gpbjnl/qzae006
DNA sequencers have become increasingly important research and diagnostic tools over the past 20 years. In this study, we developed a single-molecule desktop sequencer, GenoCare 1600 (GenoCare), which utilizes amplification-free library preparation and two-color sequencing-by-synthesis chemistry, making it more user-friendly compared with previous single-molecule sequencing platforms for clinical use. Using the GenoCare platform, we sequenced an Escherichia coli standard sample and achieved a consensus accuracy exceeding 99.99%. We also evaluated the sequencing performance of this platform in microbial mixtures and coronavirus disease 2019 (COVID-19) samples from throat swabs. Our findings indicate that the GenoCare platform allows for microbial quantitation, sensitive identification of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus, and accurate detection of virus mutations, as confirmed by Sanger sequencing, demonstrating its remarkable potential in clinical application.
Stem Cell Research & Therapy•2025•DOI: 10.1186/s13287-025-04463-7
Background: Recent advances in clinical trials have involved the transplantation of induced retinal pigment epithelium (iRPE) cells from stem cells in creating a functional monolayer that mimics the characteristics of natural adult RPE cells. One method of achieving this goal is through the use of tissue engineering. In this research, decellularised femtosecond laser intrastromal lenticules (dfLEN) were employed as a scaffold for cultivating a bioengineered iRPE monolayer sheet. Methods: iRPE cells were obtained by differentiating induced pluripotent stem cells (iPSC). These cells were then seeded on decellularized FLI-lenticules (dfLEN). The functionality, characterization, and oxidative stress of iRPE cultured on dfLEN were compared with those cultured on plates (TCP) using various assays such as immunofluorescence (IF), Edu, CCK8, ELISA, DFCH-DA, and JC-1. Additionally, RNA-seq assays and electron microscope (SEM and TEM) were used to test the iRPE characteristic on engineered dfLEN. Finally, we evaluated the biocompatibility of iRPE-dfLEN sheets by transplanting them into the subretinal space of New Zealand white rabbits. Results: The iRPE cells cultured on dfLEN exhibited morphology and physiology similar to that of native RPE tissue. The dfLEN not only increased the resistance capacity of iRPE cells but also improved their functional properties compared to TCP. In addition, our results indicate that dfLEN enhances the expression of genes associated with cilium assembly, resulting in notable improvements in ciliogenesis in iRPE cells. Finally, the dfLEN-iRPE sheets demonstrated favorable biocompatibility and some viability when transplanted into the subretinal space of rabbits for a period of 14 days.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2026058
Nucleases are a class of enzymes that specifically cleave nucleic acids in all living organisms. They play crucial roles in essential biological processes, including the regulation of gene expression, DNA damage repair, and RNA processing and degradation. MYG1 (melanocyte proliferating gene 1) is a highly conserved eukaryotic protein that exhibits 3′→5′ exonuclease activity. This study systematically characterizes the enzymatic properties of MYG1 and determines its structures in complexes with metal ions and various mono- and poly-(deoxy)nucleotides. The functional roles of key residues involved in metal ion binding and substrate binding in the catalytic reaction are examined through site-directed mutagenesis, enzymatic activity assay, and structure determination. Our biochemical and structural data together demonstrate that MYG1 is a Mn2+- or Mg2+-dependent 3′→5′ exonuclease capable of cleaving a variety of nucleic acids with different structures. It exhibits the highest activity for single-stranded RNA and a nucleotide preference for U in single-stranded RNA and dT in single-stranded DNA. Mechanistically, MYG1 functions as a dimer, with the active site formed by the catalytic domain of monomer 1 and the substrate-binding domain of monomer 2, and cleaves nucleic acids through a two-metal ion-mediated catalytic mechanism. These findings establish a molecular basis for further investigations into the biological functions and molecular mechanisms of MYG1 within cells and its potential roles in human diseases.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2026002
The newly discovered midnolin-proteasome pathway is a unique ubiquitin-independent mechanism for degrading nuclear proteins, which is crucial for maintaining cellular protein homeostasis. The Catch domain of midnolin is essential for substrate recognition and binding, yet the underlying mechanism for its broad substrate specificity remains elusive. Transcription factor IRF4, essential for the functions of B and T cells, is a substrate of midnolin. This study presents comprehensive biochemical and structural analyses of the human midnolin Catch domain in complexes with both wild-type and mutant IRF4 peptides. The crystal structure of the Catch-IRF4 complex reveals that the Catch domain creates a substrate-binding groove at the interface of the Catch1 and Catch2 subdomains, recognizing and binding to the 215-QVTGTFYAC-223 sequence motif of IRF4. The binding motif of IRF4 forms a β-strand that is embedded into the substrate-binding groove, resulting in an antiparallel five-stranded β-sheet. The interactions between the IRF4 peptide and the Catch domain are predominantly hydrophobic and exhibit high spatial complementarity. Additionally, the biochemical, modeling and structural data indicate that the V2 and A8 positions of the IRF4 sequence motif can be substituted with other hydrophobic or small polar residues (G/A/V/L/I/M/P/F/Y/C/S/T), but not with large polar and charged residues (D/N/E/Q/H/K/R). The G4 position can be replaced by Ser, while the F6 position can be substituted with Tyr. These results suggest that the Catch domain can recognize and bind to a variety of substrates containing the sequence motif x[G/A/V/L/I/M/P/F/Y/C/S/T]x[G/S]x[F/Y]x[G/A/V/L/I/M/P/F/Y/C/S/T]x or briefly the G/SxF/Y motif (where x represents polar residues) located in an unstructured or loop region on the protein surface, and the hydrophobic interactions and spatial complementarity between the binding motifs of substrates and the Catch domain govern the substrate specificity. Collectively, these findings elucidate the molecular basis for midnolin’s broad substrate specificity.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2025243
Group 2 innate lymphoid cells (ILC2s), a subset of innate lymphoid cells (ILCs) lacking antigen-specific receptors and functionally mirroring T helper 2 (Th2) cells, are indispensable components of the innate immune system that lack antigen-specific receptors but phenotypically and functionally mirror T helper 2 (Th2) cells, particularly in their expression of the transcription factor GATA3 and the secretion of type 2 cytokines for mediating type 2 immune responses. ILC2s are tissue-resident cells in mucosal tissues, including the lung, where they play crucial roles in maintaining tissue homeostasis and regulating immune responses. ILC2s are poised to respond to environmental signals such as IL-25, IL-33, and TSLP, which activate and expand ILC2s. Their functions are highly context-dependent and influenced by interactions with other immune cells. In this review, we summarize recent findings on the roles of ILC2s in lung diseases, highlighting their typical characteristics and their responsiveness to environmental signals in the context of pulmonary pathology. We also discuss potential therapeutic strategies targeting ILC2s, which may offer new avenues for the treatment of inflammatory lung diseases. Understanding the mechanisms by which ILC2s contribute to lung disease progression will provide valuable insights for the development of novel diagnostic (e.g., ILC2 phenotypic markers) and therapeutic approaches (e.g., targeting ILC2 plasticity or alarmin-ILC2 signaling axes).
Acta Biochimica et Biophysica Sinica•2024•DOI: 10.3724/abbs.2024111
This study investigates the role of lactate in the genesis and progression of ovarian cancer (OV) and explores the underlying mechanisms. Serum lactate levels show a positive correlation with tumor grade and poor prognosis in patients with OV. Bioinformatics analysis identifies CCL18 as a lactate-related gene in OV. CCL18 is up-regulated in cancerous tissues and positively related to serum lactate levels in OV patients. THP-1 cells are exposed to phorbol-12-myristate-13-acetate for M0 macrophage induction. The results of RT-qPCR and ELISA for M1/M2 macrophage-related markers and inflammatory cytokines show that the exposure of lactate to macrophages induces M2 polarization. Based on the coculture of OV cells with macrophages, lactate-treated macrophages induces a significant increase in the proliferation and migration of OV cells. However, these effects can be reversed by silencing of Gpr132 in macrophages or treatment with anti-CCL18 antibody. Experiments using the xenograft model verify that the oncogenic role of lactate in tumor growth and metastasis relies on Gpr132 and CCL18. ChIP-qPCR and luciferase reporter assays reveal that lactate regulates CCL18 expression via H3K18 lactylation. In conclusion, lactate is a potential therapeutic target for OV. It is involved in tumorigenesis by activating CCL18 expression via H3K18 lactylation in macrophages.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2025053
Krüppel-like factors (KLFs) are a family of 18 transcriptional regulators characterized by three highly conserved C2H2 zinc fingers at their C-terminal regions. KLF7, a member of this family, plays a crucial role in cell proliferation, differentiation, and the development of the nervous system, adipogenesis, diabetes, and various cancers. Studies have shown that KLF7 aggravates metabolic disorders by impeding insulin secretion and sensitivity. The nuclear factor kappa-B (NF-κB) signaling cascade is essential for inflammatory responses, while fatty acid β-oxidation is vital for metabolism. Both are linked to insulin resistance, obesity, and cardiovascular diseases. A functional link between KLF7 and the NF-κB signaling pathway has been demonstrated. In rheumatoid arthritis, KLF7 activates NF-κB signaling pathway, leading to increased cell proliferation and the production of proinflammatory cytokines, including interleukin 6 (IL-6), IL-1β, and IL-17A. In adipose tissue, KLF7 may initiate NF-κB signaling pathway by upregulating protein kinase Cζ, causing significant IL-6 secretion. KLF7 also reduces oleate-induced lipid droplets in chicken preadipocytes, indicating its role in fatty acid metabolism. Studies in mice showed that KLF7 regulates the transcription of genes of the rate-limiting glycolytic enzyme phosphofructokinase liver type (PFKL) and the fatty acid β-oxidation enzyme acyl-CoA dehydrogenase long-chain (ACADL) in cardiomyocytes independently of peroxisome proliferator-activated receptor (PPAR) γ, thereby altering heart metabolism. Additionally, KLF7 may promote cervical cancer progression by enhancing fatty acid utilization efficiency at least via ACADL upregulation. Overall, KLF7 is crucial for the regulation of fatty acid β-oxidation. KLF7 is a ring-shaped protein with zinc fingers forming the protruding part of ring surface. The lack of the third zinc finger domain in KLF7 affected its function in chicken preadipocytes. However, the importance of zinc finger domains for the regulatory function of human KLF7 remains unclear. In this study, we engineered an overexpression vector for wild-type KLF7 (pCMV-myc-KLF7_WT) and three vectors for KLF7 mutants with different zinc finger deletions (pCMV-myc-KLF7_D1, pCMV-myc-KLF7_D2, and pCMV-myc-KLF7_D3). These vectors were constructed using primers shown in Supplementary Table S1 and cDNA from HEK293T cells. Western blot analysis in the HEK293T, Ishikawa, HeLa, and EC109 cells (Pricella, Wuhan, China) showed that, unlike cells transfected with the empty vector (EV) of pCMV-myc (Clontech, Mountain View, USA), Myc-tagged proteins appeared at expected sizes in cells transfected with either the wild-type KLF7 or any of the three mutant KLF7 overexpression plasmids after 48 h (Figure 1A and Supplementary Figure S1). The impacts of overexpressing various KLF7 isoforms on gene transcription related to the NF-κB signaling pathway and fatty acid β-oxidation were evaluated using luciferase reporter assays, real-time PCR, and western blot analysis in Ishikawa, HeLa, and EC109 cells 48 h post-transfection. Details of the luciferase reporter assay transfection protocol are provided in Supplementary Table S2, the oligonucleotide sequences for real-time PCR are shown in Supplementary Table S3, and the antibodies for western blot analysis are listed in Supplementary Table S4. Compared to EV group, wild-type KLF7 overexpression significantly boosted NF-κB pathway activity in HeLa and EC109 cells (P < 0.05, Figure 1B). Furthermore, wild-type KLF7 overexpression significantly elevated IL-6 and TNF-α expressions in Ishikawa and HeLa cells (P < 0.05, Figure 1C), confirming previous findings that KLF7 enhances inflammation via the NF-κB pathway [4,5]. Cells transfected with KLF7 overexpression plasmids lacking zinc fingers showed significant differences in NF-κB pathway activities compared to those transfected with the wild-type KLF7 overexpression plasmid (P < 0.05, Figure 1B). In HeLa and EC109 cells, the absence of zinc fingers reduced NF-κB signaling activity, with the reduction proportional to the number of zinc fingers lost (P < 0.05, Figure 1B). In Ishikawa cells, losing one or two zinc fingers increased NF-κB activity compared to the wild-type KLF7 (P < 0.05, Figure 1B). Additionally, the ability of KLF7 overexpression to increase IL-6 and TNF-α expression decreased with the loss of zinc fingers.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2024212
It is unclear what part KLF7 plays in cervical cancer. In this study, immunohistochemical and bioinformatics analyses reveal that KLF7 expression is lower in normal cervical tissues than in cervical cancer tissues, and the high level of KLF7 transcripts in cervical cancer tissues is negatively correlated with patients’ overall and disease-free survival. In addition, KLF7 overexpression facilitates the proliferation, migration, and invasion of cervical cells, reduces PFKL expression, and increases the expressions of KLF4, Nanog, OCT4, CD44, SOX2, and ACADL. Additionally, knocking out the Exon 2 of KLF7 in HeLa cells results in a decrease in the total expression of KLF7 but an increase in the nuclear expression of KLF7, an increase in the capacity for proliferation, migration, invasion, and oncogenicity, and an increase in the density and ridge density of mitochondria. Consistent with these findings, RNA-seq analysis shows that knocking out the Exon 2 of KLF7 facilitates the expression of gene sets associated with cancer compared with that in wild-type HeLa cells. Moreover, the administration of alpha-lipoic acid (ALA) leads to a reduction in KLF7 expression in cells and tumor tissues, a suppression of the proliferation, migration, and invasion of HeLa and SiHa cells, and an increase in the carcinogenic potential of HeLa cells, while KLF7 overexpression shows the opposite effect on the expressions of ACADL and PFKL in HeLa and SiHa cells. In conclusion, KLF7 promotes the development of cervical cancer, and ALA can downregulate KLF7 expression and play a positive role in cervical cancer treatment.
Chinese Traditional and Herbal Drugs•2026•DOI: 10.7501/j.issn.0253-2670.2026.15.20261509
Oral ulcers are among the most prevalent inflammatory lesions of the oral mucosa, characterized by localized burning pain, dysphagia, and high recurrence rates. Conventional dressings, such as ordinary drug films, are low-cost and accessible but create a dry environment that delays epithelial cell migration and impedes healing. This study developed an amino acid-based deep eutectic solvent (DES) gel dressing loaded with curcumin, combining DES and 3D printing technology to create a localized drug delivery platform with efficient drug loading, controlled release, and excellent biocompatibility. The optimal DES system (proline-lactic acid, 1:12) increased curcumin solubility over 700-fold (0.789 mg/mL) compared to water. The optimal printing formulation and process parameters were 1% CSMA, 5% GelMA, 0.1% LAP, 0.02% tartrazine, 20% DES content, 100 μm layer height, and 25 s layer-by-layer exposure time. The resulting gel dressing exhibited excellent mechanical properties, with a 2.5 N adhesion force at 25 min and an approximately 500% swelling ratio at 6 h. In vitro biocompatibility showed L929 cell survival rates above 90%, with no significant cytotoxicity from the DES system. In an SD rat oral ulcer model, the gel achieved localized sustained curcumin release, reducing ulcer area by 40% compared to controls, and effectively decreased neutrophil density and inflammatory infiltration. The study successfully prepared a curcumin-loaded amino acid-based DES gel dressing with good wound adaptability and precise drug release behavior, offering an efficient and safe therapeutic strategy for oral ulcers. However, limitations include differences between the SD rat model and human pathophysiology, long-term biosafety concerns regarding degradation products and DES metabolism, and challenges in cost and scalability for 3D printing clinical translation.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2026002
The newly discovered midnolin-proteasome pathway is a unique ubiquitin-independent mechanism for degrading nuclear proteins, which is crucial for maintaining cellular protein homeostasis. The Catch domain of midnolin is essential for substrate recognition and binding, yet the underlying mechanism for its broad substrate specificity remains elusive. Transcription factor IRF4, essential for the functions of B and T cells, is a substrate of midnolin. This study presents comprehensive biochemical and structural analyses of the human midnolin Catch domain in complexes with both wild-type and mutant IRF4 peptides. The crystal structure of the Catch-IRF4 complex reveals that the Catch domain creates a substrate-binding groove at the interface of the Catch1 and Catch2 subdomains, recognizing and binding to the 215-QVTGTFYAC-223 sequence motif of IRF4. The binding motif of IRF4 forms a β-strand that is embedded into the substrate-binding groove, resulting in an antiparallel five-stranded β-sheet. The interactions between the IRF4 peptide and the Catch domain are predominantly hydrophobic and exhibit high spatial complementarity. Additionally, the biochemical, modeling and structural data indicate that the V2 and A8 positions of the IRF4 sequence motif can be substituted with other hydrophobic or small polar residues (G/A/V/L/I/M/P/F/Y/C/S/T), but not with large polar and charged residues (D/N/E/Q/H/K/R). The G4 position can be replaced by Ser, while the F6 position can be substituted with Tyr. These results suggest that the Catch domain can recognize and bind to a variety of substrates containing the sequence motif x[G/A/V/L/I/M/P/F/Y/C/S/T]x[G/S]x[F/Y]x[G/A/V/L/I/M/P/F/Y/C/S/T]x or briefly the G/SxF/Y motif (where x represents polar residues) located in an unstructured or loop region on the protein surface, and the hydrophobic interactions and spatial complementarity between the binding motifs of substrates and the Catch domain govern the substrate specificity. Collectively, these findings elucidate the molecular basis for midnolin’s broad substrate specificity.
Acta Biochimica et Biophysica Sinica•2026•DOI: 10.3724/abbs.2026058
Nucleases are a class of enzymes that specifically cleave nucleic acids in all living organisms. They play crucial roles in essential biological processes, including the regulation of gene expression, DNA damage repair, and RNA processing and degradation. MYG1 (melanocyte proliferating gene 1) is a highly conserved eukaryotic protein that exhibits 3′→5′ exonuclease activity. This study systematically characterizes the enzymatic properties of MYG1 and determines its structures in complexes with metal ions and various mono- and poly-(deoxy)nucleotides. The functional roles of key residues involved in metal ion binding and substrate binding in the catalytic reaction are examined through site-directed mutagenesis, enzymatic activity assay, and structure determination. Our biochemical and structural data together demonstrate that MYG1 is a Mn2+- or Mg2+-dependent 3′→5′ exonuclease capable of cleaving a variety of nucleic acids with different structures. It exhibits the highest activity for single-stranded RNA and a nucleotide preference for U in single-stranded RNA and dT in single-stranded DNA. Mechanistically, MYG1 functions as a dimer, with the active site formed by the catalytic domain of monomer 1 and the substrate-binding domain of monomer 2, and cleaves nucleic acids through a two-metal ion-mediated catalytic mechanism. These findings establish a molecular basis for further investigations into the biological functions and molecular mechanisms of MYG1 within cells and its potential roles in human diseases.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21345
BACKGROUND: Periodontitis is a chronic inflammatory disease. Previous research has predominantly focused on specific immune cells or cytokines. Therefore, systematically elucidating its immune mechanisms and discovering novel therapeutic targets hold significant implications. OBJECTIVE: To analyze the expression profiles of periodontitis-associated immune cell subpopulations and identify key differentially expressed genes with a causal relationship to the disease, thereby exploring potential molecular mechanisms and key genes involved in periodontitis and immune cell dynamics. METHODS: Single-cell RNA sequencing data from the GEO database were used to analyze immune cell subset heterogeneity and identify differentially expressed genes. Mendelian randomization analysis was performed using expression quantitative trait loci data to infer causal relationships between immune cell gene expression and periodontitis risk. Pathway enrichment and immune infiltration analyses were performed on the identified causal genes to reveal the associations between differentially expressed genes and immune cells with the development and progression of periodontitis. CellChat trajectory analysis was used to explore intercellular communication. To validate key findings, gingival tissue samples were collected from 20 patients with periodontitis diagnosed by the Department of Stomatology at The First Affiliated Hospital of Shihezi University (periodontitis group) and 20 healthy gingival tissue samples from patients undergoing orthodontic or impacted tooth extraction (control group). RT-qPCR and immunohistochemistry were used to detect the expression of key genes. RESULTS AND CONCLUSION: Comprehensive analysis identified 23 immune cell clusters in periodontitis and three key genes with significant causal relationships to periodontitis risk: annexin A1 (ANXA1), solute carrier family 11 member 1 (SLC11A1), and vimentin (VIM). Pathway enrichment analysis revealed their involvement in key immune regulatory mechanisms. Further analyses of immune subtype receptor-ligand interactions and key cell subtype trajectories characterized the distinct roles of ANXA1, SLC11A1, and VIM in disease progression. Compared with healthy controls, the mRNA expression levels of ANXA1, SLC11A1, and VIM were upregulated in periodontitis tissues (P < 0.05). This study reveals the key roles of immune cell subpopulations in periodontitis and validates causal genes (ANXA1, SLC11A1, VIM) associated with the disease.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21378
BACKGROUND: Previous studies have demonstrated that Gugutou Huaisiyu Capsule effectively alleviate pain in patients with osteonecrosis of femoral head; however, the mechanism of action remains unclear. OBJECTIVE: To investigate the mechanism by which Gugutou Huaisiyu Capsule mitigate peripheral pain sensitization in rats with steroid-induced osteonecrosis of the femoral head. METHODS: Thirty-two Sprague-Dawley rats were randomly assigned to four groups: blank control, model, low-dose Gugutou Huaisiyu Capsule, and high-dose Gugutou Huaisiyu Capsule groups with eight rats in each group. Steroid-induced femoral head necrosis rat models were established via intraperitoneal injection of lipopolysaccharide combined with gluteal injection of methylprednisolone in the later three groups. After 7 days of modeling, the low- and high-dose Chinese medicine groups were administered 0.33 and 0.67 g/kg of Gugutou Huaisiyu Capsule decoction by gavage, respectively, while the blank control and model groups received normal saline, once daily for 8 weeks. After the last administration, Micro-CT was used to analyze the microstructure of the femoral head cancellous bone; Von-Frey test was used to detect mechanical pain threshold; ELISA was used to detect serum levels of pain mediators calcitonin gene-related peptide (CGRP) and substance P; multiplex immunofluorescence staining and real-time quantitative PCR were used to detect protein and gene expression of tyrosine kinase receptor A (TrkA), CGRP, and neuronal marker β3-tubulin in the femoral head and dorsal root ganglion. RESULTS AND CONCLUSION: (1) Micro-CT and pain behavior tests showed that compared with the blank control group, the model group had significant destruction of the femoral head cancellous bone microstructure, decreased bone mineral density and bone volume fraction, increased trabecular separation, and decreased mechanical pain threshold. Compared with the model group, the low- and high-dose Chinese medicine groups had significantly improved trabecular structure, increased bone mineral density and bone volume fraction, decreased trabecular separation, and increased mechanical pain threshold. (2) ELISA showed that the levels of CGRP and substance P in the model group were significantly higher than those in the blank control group, while the levels in the low- and high-dose Chinese medicine groups were lower than those in the model group. (3) Multiplex immunofluorescence staining and real-time quantitative PCR showed that compared with the blank control group, the protein and gene expression of CGRP, TrkA, and β3-tubulin in the femoral head and dorsal root ganglion were increased in the model group; compared with the model group, the expression of these proteins and genes was decreased in the low- and high-dose Chinese medicine groups. (4) These results indicate that Gugutou Huaisiyu Capsule can increase the mechanical pain threshold, inhibit abnormal neuronal activation, and alleviate pain sensitization in rats with steroid-induced osteonecrosis of the femoral head, and the mechanism may be related to the inhibition of TrkA.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21502
OBJECTIVE: The therapeutic modalities of physical factor interventions for knee osteoarthritis have been increasingly diversified; however, comprehensive comparative evaluations of their efficacy remain limited. This study aims to compare the efficacy and safety of various physical factor therapies for knee osteoarthritis through a network meta-analysis. METHODS: Randomized controlled trials on physical factor therapy for knee osteoarthritis were retrieved from PubMed, Web of Science, Cochrane Library, EMbase, CNKI, VIP, Wanfang, and CBM databases from inception to July 25, 2025. After literature screening and data extraction, the quality of included studies was assessed using the Cochrane risk-of-bias tool. Statistical analyses were performed using Stata 16.0 and RevMan 5.4.1. RESULTS: A total of 65 studies involving 3,418 patients (1,726 in treatment groups, 1,692 in control groups) were included, covering seven physical factor therapies. Network meta-analysis showed that for improving total effective rate, the top three interventions by surface under the cumulative ranking curve (SUCRA) were pulsed electromagnetic field + conventional rehabilitation, ultrasound + conventional rehabilitation, and transcutaneous electrical stimulation + conventional rehabilitation. For improving visual analogue scale (VAS) score, the top three were transcutaneous electrical stimulation + conventional rehabilitation, extracorporeal shock wave + conventional rehabilitation, and ultrasound + conventional rehabilitation. For improving WOMAC total score, the top three were ultrasound + conventional rehabilitation, pulsed electromagnetic field + conventional rehabilitation, and ultrasound + transcutaneous electrical stimulation + conventional rehabilitation. For reducing WOMAC stiffness score, the top three were transcutaneous electrical stimulation + conventional rehabilitation, ultrasound + conventional rehabilitation, and ultrasound + transcutaneous electrical stimulation + conventional rehabilitation. For improving SF-36 quality of life score, the top three were pulsed electromagnetic field + conventional rehabilitation, extracorporeal shock wave + conventional rehabilitation, and ultrasound + conventional rehabilitation. For improving Lysholm knee score, the top three were ultrasound + conventional rehabilitation, ultrasound + transcutaneous electrical stimulation + conventional rehabilitation, and extracorporeal shock wave + conventional rehabilitation. Regarding adverse events, no serious adverse events were reported; most studies reported only mild skin irritation or allergic reactions. CONCLUSION: Transcutaneous electrical stimulation combined with conventional rehabilitation showed superior advantages in improving VAS and WOMAC stiffness scores; ultrasound combined with conventional rehabilitation performed relatively better in improving Lysholm knee score and WOMAC total score; pulsed electromagnetic field combined with conventional rehabilitation had potential advantages in improving overall quality of life. Each physical factor has its unique advantages, but limited by the quality and quantity of included studies, these conclusions need to be verified by more high-quality, multi-center, large-sample randomized controlled trials.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21577
BACKGROUND: Recent studies have shown that Chlorella possesses potential value in treating rheumatoid arthritis. The pathological progression of rheumatoid arthritis is closely associated with an imbalance in oxidative stress, abnormal macrophage polarization, aggressive activation of fibroblast-like synoviocytes, and disturbances in the vascular endothelial system. However, the optimization of extraction processes for peptides derived from Chlorella and their regulatory effects on key pathological links of rheumatoid arthritis remain to be systematically validated. OBJECTIVE: To optimize the extraction process of antioxidant peptides from Chlorella, clarify their antioxidant activity and biosafety, and explore their regulatory effects on pathological phenotypes of rheumatoid arthritis-related cells (RAW 264.7 mouse monocyte macrophage leukemia cells, fibroblast-like synoviocytes, and human umbilical vein endothelial cells), providing experimental evidence for the treatment of rheumatoid arthritis with Chlorella peptides. METHODS: (1) Chlorella peptide extract was prepared by bromelain enzymatic hydrolysis combined with phosphomolybdic acid precipitation. Using peptide yield as the evaluation index, the extraction process parameters were optimized by single-factor experiments, including solid-liquid ratio, enzymatic hydrolysis time, and reaction system pH. (2) The peptide content was determined by BCA method, antioxidant capacity was detected by ABTS method, and biosafety of peptides on RAW 264.7 cells, fibroblast-like synoviocytes, and human umbilical vein endothelial cells was evaluated by CCK-8 method. (3) An inflammatory model of RAW 264.7 cells induced by lipopolysaccharide was established. The effects of peptides on intracellular reactive oxygen species levels and M1/M2 polarization phenotypes were detected by DCFH-DA staining, flow cytometry, and real-time fluorescence quantitative reverse transcription polymerase chain reaction. (4) An activation model of fibroblast-like synoviocytes induced by tumor necrosis factor-alpha was established. The effects of peptides on migration, proliferation, invasion, and related gene expression of fibroblast-like synoviocytes were detected by wound healing assay, EdU proliferation assay, Transwell invasion assay, and real-time fluorescence quantitative reverse transcription polymerase chain reaction. (5) An abnormal activation model of human umbilical vein endothelial cells induced by vascular endothelial growth factor A was established. The effects of peptides on migration, tube formation, and expression of hypoxia-inducible factor 1 alpha and vascular endothelial growth factor A genes were detected by wound healing assay, Transwell assay, tube formation assay, and real-time fluorescence quantitative reverse transcription polymerase chain reaction. RESULTS AND CONCLUSION: (1) The optimal extraction process for Chlorella peptides was solid-liquid ratio of 2:1 (g:100 mL), enzymatic hydrolysis time of 60 minutes, and reaction system pH of 6.5, yielding the highest peptide yield. (2) Chlorella peptides exhibited concentration-dependent antioxidant activity and showed no obvious toxicity to RAW 264.7 cells, fibroblast-like synoviocytes, and human umbilical vein endothelial cells in the concentration range of 1-10 μg/mL, indicating good biocompatibility. (3) Chlorella peptides dose-dependently inhibited lipopolysaccharide-induced reactive oxygen species generation in RAW 264.7 cells, downregulated M1 pro-inflammatory genes such as interleukin-1 beta and tumor necrosis factor-alpha, upregulated M2 anti-inflammatory genes such as interleukin-10 and arginase 1, and promoted macrophage polarization from M1 to M2 phenotype. (4) Chlorella peptides significantly inhibited tumor necrosis factor-alpha-induced migration, proliferation, and invasion of fibroblast-like synoviocytes, and downregulated the expression of interleukin-6, matrix metalloproteinase 13, tumor necrosis factor receptor superfamily member 11A, and C-X-C motif chemokine ligand 12. (5) Chlorella peptides effectively inhibited vascular endothelial growth factor A-induced migration and tube formation of human umbilical vein endothelial cells, and reduced the expression of hypoxia-inducible factor 1 alpha and vascular endothelial growth factor A genes. These results indicate that Chlorella peptides regulate multiple pathological links of rheumatoid arthritis through anti-oxidative stress, regulation of macrophage polarization, inhibition of aggressive phenotype of fibroblast-like synoviocytes, and improvement of vascular endothelial disorders, suggesting potential therapeutic value for rheumatoid arthritis.
Chinese Journal of Tissue Engineering Research•2026•DOI: 10.12307/2026.21575
BACKGROUND: For diabetic foot infections, traditional vancomycin-loaded bone cement has a limited antibacterial spectrum, and there is an urgent need for novel drug carriers. OBJECTIVE: To explore the in vitro elution characteristics of polymyxin B sulfate-loaded bone cement. METHODS: Polymyxin B sulfate powder was uniformly mixed with polymethyl methacrylate bone cement and poured into molds to prepare bone cement microspheres with diameters of 5 and 7 mm. The two types of bone cement microspheres of different diameters were immersed in 1 mL of PBS, and elution samples were collected at specific time points. The concentration of polymyxin B sulfate in the eluent was determined by mass spectrometry, and the drug release pattern was analyzed. RESULTS AND CONCLUSION: (1) The drug release peaks for both types of bone cement microspheres occurred between 0 and 0.5 h, after which the release rate gradually decreased. Inter-group comparison showed that the drug release rate and cumulative release rate of the 5 mm diameter microspheres were higher than those of the 7 mm diameter microspheres. The cumulative drug release rates at 14 days for the 5 mm and 7 mm diameter microspheres were 5.08% and 3.37%, respectively. The drug release from both types of bone cement microspheres of different diameters reached 90% of the total release within 7 days, approaching the release endpoint. The in vitro drug release curves of both types of microspheres conformed to the Ritger-Peppas model (R2=0.998 56, 0.990 90), with Fickian diffusion as the dominant mechanism. (2) The polymyxin B sulfate-loaded bone cement exhibited sustained release characteristics, with drug release mainly concentrated in the first 7 days and low release thereafter. Therefore, 5-7 days after implantation is the optimal timing for secondary debridement; continued retention poses a higher risk of inducing bacterial resistance. The drug release rate is related to the size of the bone cement, and in clinical application, bone cement microspheres with a diameter of about 5 mm can be prioritized to balance rapid drug release and long-term antibacterial needs.
Stem Cell Research & Therapy•2026•DOI: 10.1186/s13287-026-04986-7
Tonsil mesenchymal stem cells (TMSCs) represent a promising source for regenerative medicine, yet their clinical translation is constrained by replicative senescence during in vitro expansion. This study investigated senescence-associated phenotypic changes and underlying regulatory mechanisms in human palatine tonsil-derived MSCs. TMSCs were isolated and characterized, then serially passaged to early (P1–P5) and late (beyond P10) stages. Proliferation declined progressively with passage, as assessed by CCK-8. Senescence-associated β-galactosidase (SA-β-gal) staining revealed a significantly higher percentage of positive cells in late-passage TMSCs. Protein levels of p16, p53, and p21 were markedly upregulated in aged cells. RNA sequencing identified differentially expressed genes (DEGs) between young and senescent TMSCs, with KEGG enrichment highlighting the PI3K-Akt signaling pathway, ECM-receptor interaction, and calcium signaling. Western blot confirmed a significantly increased p-Akt/Akt ratio in senescent TMSCs. These findings establish that replicative senescence in TMSCs is associated with activation of the PI3K-Akt pathway, which likely orchestrates senescence through p16 and p53-p21 cascades. The results provide mechanistic insights into stem cell aging and suggest potential molecular targets for delaying TMSC senescence in regenerative applications.
Acta Biochimica et Biophysica Sinica•2025•DOI: 10.3724/abbs.2025053
Krüppel-like factor 7 (KLF7) is a C2H2 zinc-finger transcriptional regulator implicated in insulin resistance, inflammation, and fatty acid metabolism. The functional necessity of its three zinc fingers for human KLF7 activity remained undefined. We engineered pCMV-myc-KLF7_WT and three zinc-finger deletion mutants (D1, D2, D3) and expressed them in HEK293T, Ishikawa, HeLa, and EC109 cells. Western blot confirmed expected Myc-tagged protein sizes at 48 h. Luciferase reporter assays showed that wild-type KLF7 significantly increased NF-κB activity (P < 0.05) and the promoter activities of ACADL, ECH1, and HADHB (P < 0.05) across all three cell lines, whereas deletion of any single zinc finger abolished these effects (P < 0.05). KLF7 did not alter NFKB1 mRNA (P > 0.05) or RelA (P65) protein levels, indicating downstream action within the NF-κB cascade. For fatty acid β-oxidation, wild-type KLF7 elevated CPT1A, ACADL, and ECH1 promoter activities and protein expression, with ACTB as internal control; zinc-finger-deficient mutants failed to sustain these transcriptional and translational increases. These data establish that the zinc-finger domain is indispensable for KLF7-mediated enhancement of NF-κB signaling and fatty acid β-oxidation gene transcription in human cells, providing a mechanistic basis for targeting KLF7 zinc fingers in metabolic and inflammatory disorders.