Zhihong Wu

TBX6-associated congenital scoliosis (TACS) is a genetic subtype of congenital scoliosis caused by reduced TBX6 gene dosage, most commonly through compound inheritance of a rare loss-of-function allele and a hypomorphic T-C-A haplotype. However, the mechanisms linking TBX6 dysfunction to impaired human skeletal development remain unclear. Here, we established a patient-derived induced pluripotent stem cell (iPSC) model of TACS and recapitulated key stages of axial skeletal development through stepwise differentiation into paraxial mesoderm, nascent somites, sclerotome, and subsequently osteogenic and chondrogenic lineages. TACS-derived cells showed impaired differentiation, with reduced mineralized matrix deposition and diminished cartilage matrix formation. Bulk transcriptomic profiling revealed marked alterations in both lineages. Notably, mu-cin-type O-glycan biosynthesis was consistently enriched among downregulated genes in osteogenic and chondrogenic cells and was also significantly represented among genes commonly downregulated across both lineages. qRT-PCR further confirmed reduced expression of GALNT3, GALNT5, GALNT7, GALNT10, GCNT1, and ST6GALNAC2, which participate in distinct steps of mucin-type O-glycan biosynthesis. Consistent with these transcriptional alterations, Tn-antigen staining showed reduced Tn-associated O-GalNAc signals in TACS-derived osteogenic and chondrogenic cells. In silico promoter analysis identified candidate sequences matching the TBX6-binding motif within the upstream regions of these genes, providing potential regulatory regions for future experimental investigation. Collectively, these results establish a human iPSC model for TACS and identify coordinated dysregulation of a mucin-type O-glycan biosynthesis-related transcriptional program, accompanied by reduced Tn-associated O-GalNAc signals, as molecular features associated with impaired osteochondral differentiation in this model. These findings should be interpreted within the context of a single family-based iPSC model, and validation in additional TACS lines and isogenic TBX6-corrected models will be required.

Wearable technologies are reshaping gait rehabilitation by shifting assessment and therapy from intermittent, clinic-based observation to continuous, data-driven, and adaptive care. This narrative review synthesizes the biomechanical basis of gait instability, the architecture and classification of wearable rehabilitation systems, and the clinical evidence for motion sensors, smart insoles, biofeedback devices, robotic and orthotic wearables, neuromodulatory systems, immersive platforms, and artificial intelligence (AI)-enabled closed-loop interventions. Its main contribution is an integrated framework that links AI, digital biomarkers, device components, adaptive control, and translational implementation, rather than treating wearable rehabilitation as a device-only or disease-specific topic. Current evidence indicates that these technologies can improve gait speed, symmetry, balance, endurance, fall-risk monitoring, and dual-task performance in neurological, musculoskeletal, frailty-related, and aging populations. However, the field is still limited by heterogeneous protocols, small samples, limited longitudinal validation, insufficient device standardization, usability barriers, cybersecurity concerns, uncertain reimbursement, and restricted interoperability with healthcare systems. Future progress will depend on multimodal sensor fusion, explainable and federated AI, digital twins, adaptive wearable robotics, tele-rehabilitation pathways, and large-scale pragmatic trials that validate effectiveness in real-world rehabilitation settings.

Critical‐sized bone defects remain a major clinical challenge in orthopedics and regenerative medicine. Bone organoid provided therapy potentials; however, current engineering strategies generally neglect the bioenergetic foundation and intrinsic mechano‐electrical coupling microenvironment of native bone tissue. Herein, we prepared a three‐dimensional native bone electromechanical niche‐mimicking mechano‐electrical coupling scaffold based on piezoelectric potassium sodium niobate and methacrylated gelatin. Experiments demonstrate that this scaffold establishes an enabling physiologically relevant mechanical–electrical microenvironment throughout organoid construction under dynamic mechanical stimulation, which markedly enhances osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs)and simultaneously endows the engineered organoids with robust pro‐angiogenic and pro‐lymphangiogenic paracrine functions, achieving efficient coordinated regeneration in a rat critical‐sized calvarial defect. RNA‐sequencing together with fluorescent staining proved that mechano‐electrical coupling signals activate the PIEZO1 channel to promote Ca ²⁺ influx into the cytoplasm and simultaneously upregulate mitochondrial calcium uniporter (MCU) expression, thereby enhancing mitochondrial Ca ²⁺ . The increased mitochondrial Ca ^{2 +} enrichment promoted tricarboxylic acid cycle enzyme activities and elevated cellular ATP production, which efficiently supports the functional maturation of bone organoids. Overall, this work establishes a bioenergetic engineering construction strategy for bone organoids with enhanced vascular/lymphatic system regeneration, providing both a conceptual framework and an engineering approach for organoid functional optimization and critical‐sized bone defect regenerative treatment.

Extracellular vesicles (EVs) derived from human umbilical cord mesenchymal stem cells (hUCMSCs) have emerged as promising therapeutic candidates for osteoarthritis (OA). Sirtuin 6 (SIRT6), a class III histone deacetylase, exerts protective effects by preventing cellular senescence, reducing inflammation, and promoting longevity. This study aims to generate immortalized hUCMSCs with SIRT6 and telomerase reverse transcriptase (TERT) overexpression and investigate the therapeutic potential of their secreted EVs (EVs@SIRT6) in OA. hUCMSCs were genetically engineered using lentiviral transduction to overexpress SIRT6 and TERT, referred to as SIRT6/TERT-hUCMSCs. Natural EVs and EVs@SIRT6 were isolated via differential ultracentrifugation and characterized by morphology, size distribution, marker expression, and proteomic signature. Their effects on IL-1β stimulated chondrocytes were assessed in vitro, including cell viability, apoptosis, scratch closure, inflammatory cytokine secretion, and oxidative stress. Transcriptomic alterations were analyzed by RNA sequencing. Therapeutic efficacy in vivo was evaluated in a rat anterior cruciate ligament transection (ACLT) model via micro-computed tomography, histological analyses, and immunohistochemistry. SIRT6/TERT-hUCMSCs preserved mesenchymal identity, trilineage differentiation potential, and sustained proliferative capacity up to passage 60, accompanied by longer telomeres and reduced senescence compared to parental cells. EVs@SIRT6 maintained typical EV features but were enriched in SIRT6 protein and displayed a distinct proteomic signature, comprising 1,440 unique proteins associated with nuclear/chromatin repair. In vitro, EVs@SIRT6 more effectively enhanced chondrocyte viability and wound healing while reducing apoptosis compared with natural EVs. EVs@SIRT6 also markedly alleviated inflammatory response, promoted anabolism, suppressed catabolism, and mitigated oxidative stress in chondrocytes. The protective effects of EVs@SIRT6 on chondrocytes were associated with the suppression of the JAK-STAT signaling pathway. Studies in a rat ACLT model further confirmed that EVs@SIRT6 outperformed natural EVs in attenuating subchondral osteosclerosis, reducing osteophyte formation, and mitigating cartilage damage. Engineered EVs@SIRT6 outperform natural EVs in preserving chondrocytes homeostasis and reducing OA progression, establishing an efficient platform for preparing engineered EVs for clinical application.

Background: Although auricular acupoint‐related therapy (AART) is increasingly used for postoperative rehabilitation after total knee arthroplasty (TKA), evidence comparing the effectiveness of different AART modalities remains limited. This study aimed to evaluate the efficacy and safety of AART through a systematic review and network meta‐analysis (NMA). Methods: We performed a systematic review and NMA of randomized controlled trials (RCTs) from multiple Chinese and English databases, from inception to June 2026. Two reviewers independently screened studies, extracted data, and assessed risk of bias using the Cochrane ROB‐2 tool and evidence quality with GRADE. Pairwise comparisons and NMA were conducted using RevMan 5.4 and R software. Results: Twenty‐four RCTs involving 1790 patients were included. Compared with routine rehabilitation, AART significantly reduced resting pain scores on postoperative day 1 (mean difference [MD] = −0.38) and day 3 (MD = −0.61) and improved movement‐evoked pain on days 3 and 7. AART also enhanced knee function (Hospital for Special Surgery scores), increased active range of motion, and reduced complications such as vomiting, dizziness, and urinary retention. NMA suggested that auricular acupressure combined with Chinese medicine transdermal therapy may be the most effective intervention for early resting pain. Of note, the certainty of evidence for most outcomes was low or very low, and the findings should be interpreted with caution in clinical practice, serving only as preliminary reference rather than definitive clinical guidance. Conclusion: AART may serve as an effective non‐pharmacological adjunct in multimodal rehabilitation after TKA, particularly for acute pain relief, functional recovery, and complication reduction. High‐quality trials are needed to confirm these findings.

This study investigates the complex interplay between gut microbiome dysbiosis and systemic inflammation as a critical risk factor in the pathogenesis of osteoarthritis (OA). Furthermore, it elucidates the role of gut microbiota (GMB) dysbiosis in driving OA progression. A refined “double-hit” murine model was developed to explore this relationship. The first intervention involved inducing gut microbiota dysbiosis through the administration of colistin and Escherichia coli, followed by surgical destabilization of the medial meniscus (DMM) to induce joint instability. The composition of the gut microbiota was analyzed using 16 S rRNA sequencing. Gut permeability was assessed via RT-PCR and immunofluorescence (IF), while mRNA sequencing was employed to examine alterations in gene expression. Treatment with colistin and E. coli significantly altered the gut microbiota composition, characterized by a marked increase in the absolute abundance of Firmicutes and a concomitant reduction in Bacteroidota and the Bacteroidota/Firmicutes (B/F) ratio. At the genus level, the absolute abundances of Muribaculaceae, Rikenellaceae_RC9_gut_group, and Roseburia were significantly diminished. GMB dysbiosis led to the downregulation of intestinal tight junction proteins, including ZO-1 and Occludin, resulting in increased intestinal permeability. Consequently, serum levels of lipopolysaccharide (LPS) were significantly elevated, indicating LPS translocation from the gut into systemic circulation. Notably, GMB dysbiosis markedly exacerbated OA progression, as evidenced by accelerated cartilage degeneration, increased osteophyte formation, and reduced bone mineral density (BMD). The OARSI scoring system revealed that OA severity in both colistin and E. coli treatment groups was significantly higher than in the control group. Additionally, GMB dysbiosis promoted the expression of inflammation-related genes in the synovium and induced M1 polarization of macrophages, demonstrated by the upregulation of CD86 and an elevated CD86/CD206 ratio. Correlation analyses indicated that Bacteroidota and the B/F ratio were positively associated with intestinal barrier integrity and negatively correlated with OA progression. In contrast, Firmicutes exhibited a positive correlation with inflammation and OA deterioration. These findings collectively underscore the critical role of GMB dysbiosis in modulating intestinal permeability, systemic inflammation, and OA pathogenesis. The protective effects of Bacteroidota and the B/F ratio, as well as the detrimental impact of Firmicutes, highlight potential therapeutic targets for mitigating OA progression through GMB modulation.

Background and Objective: Glypican-5 (GPC5) has been well-characterized as a tumor suppressor in lung adenocarcinoma (LUAD); however, the functional implications of its germline mutations in cancer pathogenesis remain largely unexplored. In this study, we identified and characterized a pathogenic GPC5 variant (c.776C>T, p.Pro259Leu) within a Chinese LUAD pedigree, systematically investigating its oncogenic mechanisms through comprehensive molecular and cellular analyses.Methods: Our investigation employed a multifaceted approach beginning with the recruitment of a LUAD-affected family cohort (n=4 patients, 1 healthy control), followed by exome sequencing of matched blood and FFPE tumor samples. Through rigorous rare variant analysis, we prioritized the GPC5 c.776C>T variant, subsequently validating its pathogenicity via integrated computational modeling and immunohistochemical profiling. Mechanistic studies in A549 and H2009 LUAD cell lines encompassed: (1) comprehensive proliferation and apoptosis assessment using CCK-8, colony formation, EdU incorporation, and flow cytometry; (2) migration and invasion evaluation through Transwell and wound healing assays; (3) EMT/Wnt pathway interrogation via Western blot analysis of E-cadherin, N-cadherin, Vimentin, and β-catenin expression patterns; and (4) definitive functional validation through GPC5 overexpression and knockdown experiments.Results: Genetic analysis revealed the GPC5 c.776C>T variant exhibited complete cosegregation with LUAD phenotype in the pedigree while being absent in control populations (gnomAD frequency: 0.000003989), accompanied by significantly reduced GPC5 expression in tumor tissues. Functional characterization demonstrated that compared to wild-type, the mutant variant conferred aggressive oncogenic properties: significantly enhanced proliferative capacity, impaired apoptosis induction, and markedly increased migratory potential. Molecular analyses revealed the mutant promoted EMT activation through nuclear β-catenin accumulation and subsequent upregulation of mesenchymal markers. Crucially, siRNA-mediated GPC5 knockdown phenocopied these oncogenic effects, providing definitive evidence of its tumor-suppressive function.Discussion: Our findings establish that the GPC5 c.776C>T mutation drives LUAD progression through a novel molecular mechanism involving impaired β-catenin degradation, subsequent nuclear translocation, and consequent EMT activation. These results position GPC5 as a critical nodal regulator of Wnt/β-catenin signaling in LUAD pathogenesis and suggest its germline mutations may serve as valuable biomarkers for hereditary LUAD risk assessment. Therapeutically, these findings highlight the potential utility of Wnt pathway inhibitors in managing GPC5-mutant LUAD cases, while also providing a molecular framework for future investigations into glypican family members in cancer biology.

Purpose: Hepatocellular carcinoma (HCC) is one of the most common cancers and a leading cause of death worldwide. Accurate prognosis prediction tools are urgently needed. While the use of circulating tumor cells (CTCs) as prognostic prediction tool has a clear potential. Methods: We established a comprehensive, negative enrichment-based strategy for CTCs analysis in patients with HCC, involving identification of epithelial CTCs (E-CTCs) and mesenchymal CTCs (M-CTCs) through specific biomarker. This strategy was performed in 127 HCC cases, 21 nonmalignant liver disease (NMLD) patients and 42 health control to analyze the relevance between CTCs and tumor recurrence. Results: The total CTC number and M-CTC percent were positively correlated with tumor malignancy and high recurrence risk. Individually, preoperative total CTC number and M-CTC percent could robustly distinguish relapse cases from those with no relapse, with sensitivity of 80.95% and 90.48%, specificity of 74.12% and 84.71%, respectively. Levels of preoperative total CTC number and M-CTC percent can both be regarded as independent risk factors for HCC with early recurrence (P = 0.0053, P < 0.0001), and are both significantly correlated with worse recurrence-free survival (RFS) (log rank P < 0.0001; HR 7.78, 95% CI = 3.59–16.87; log rank P < 0.0001; HR 24.4, 95% CI = 8.67–68.77). The levels of total CTC number and M-CTC number had higher effectiveness than alpha fetal protein (AFP) in HCC longitudinal supervision (77.78% vs 88.89% vs 22.22%). Conclusion: Preoperative and postoperative CTCs with higher effectiveness than AFP in prognosis prediction and recurrence supervision, indicating that CTCs could work as the biomarker for HCC clinical management.