Zhoushan Hospital

Variants in OTUD5 are associated with neurodevelopmental disorders (NDDs), yet the underlying molecular mechanisms remain unclear. This study aimed to investigate the pathogenicity of a novel OTUD5 variant (c.697G > A, p.Val233Met) and elucidate its regulatory role in neural progenitor cell (NPC) proliferation and differentiation, thereby uncovering the function of OTUD5 in neurodevelopment. The OTUD5 variant was identified in two NDD patients via exome sequencing. Patient-derived induced pluripotent stem cells (iPSCs) and CRISPR/Cas9-corrected isogenic controls were generated. NPC proliferative activity was assessed by Ki67 immunofluorescence staining, cell-cycle distribution was analyzed by flow cytometry, and neuronal differentiation was evaluated by Tuj1/MAP2 immunofluorescence staining. Substrate screening was conducted in HEK293T cells using co-immunoprecipitation (Co-IP) and mass spectrometry. Deubiquitination capacity and protein stability were validated through ubiquitination assays and cycloheximide (CHX) chase experiments. The p.Val233Met variant, located within the catalytic OTU domain, induced a marked conformational alteration in the OTUD5 protein. Functionally, the variant caused aberrant NPC proliferation (1.8-fold increase in Ki67+ cells, accompanied by release of G1 arrest) and impaired neuronal differentiation (60% reduction in Tuj+ cells). Mechanistically, wild-type OTUD5 stabilized GSK3β by removing K48-linked ubiquitin chains, whereas the mutant isoform exhibited diminished deubiquitinase activity, accelerating GSK3β degradation and shortening its half-life by 40%. This study establishes a novel disease mechanism whereby OTUD5 mutations disrupt NPC homeostasis through GSK3β destabilization, highlighting the critical role of ubiquitination regulation in neurodevelopment. Our iPSC model provides a platform for testing GSK3β-targeted therapies in OTUD5-related NDDs.

BackgroundLow-Dose Computed Tomography (LDCT) is commonly used to detect pulmonary nodules; however, it may also contribute to overdiagnosis. DNA methylation shows promise as an approach to discriminate early-stage lung cancer (LC) patients from individuals with benign nodules (hereafter referred to as benign pulmonary nodule group) and healthy individuals.MethodsThis study investigated the performance of DNA methylation in three genes (TAC1, SOX17, and RASSF1A) in plasma-derived cell-free DNA (cfDNA) for discriminating LC patients from benign and healthy individuals (collectively referred to as benign/healthy individuals). We enrolled 149 LC patients (96 with early-stage [stage IA] and 53 with advanced-stage [non-IA]), 54 benign pulmonary nodule group, and 75 healthy individuals.ResultsMethylation-positive rates for all three genes were significantly higher in LC patients compared to benign/healthy individuals. A combined three-gene model based on the ΔCt values of the three genes demonstrated robust diagnostic performance, achieving a sensitivity of 97.7%, specificity of 96.6%, and an area under the curve (AUC) of 0.99 for discriminating LC patients from benign/healthy individuals. Furthermore, another combined three-gene model based on the ΔCt values of the same genes showed high diagnostic performance for discriminating IA-stage LC patients from benign/healthy individuals, with a sensitivity of 96.9%, specificity of 88.54%, and AUC of 0.95.ConclusionThis study highlights the robust diagnostic value of a combined three-gene (TAC1, SOX17, and RASSF1A) methylation model for detecting LC, including early-stage disease, offering high sensitivity and specificity.

Background Abdominal aortic aneurysm (AAA) is a progressive vascular disorder that enlarges irreversibly and may ultimately rupture. Mendelian randomization (MR) provides a powerful approach to uncover biomarkers. We aimed to identify plasma proteins linked to AAA and evaluate their diagnostic potential. Methods We performed a proteome-wide MR analysis using genetic instruments for 2,940 plasma proteins from the UK Biobank Pharma Proteomics Project (UKB-PPP). Summary statistics for AAA were obtained from the Finnish R9 GWAS, comprising 4,083 cases and 420,324 controls. Results Eleven proteins showed significant causal associations with AAA risk (FDR < 0.05): seven positively and four negatively associated, highlighting their potential as diagnostic markers. Conclusion This study provides the first proteome-wide MR evidence linking plasma proteins to AAA, offering candidate biomarkers for diagnosis. As a hypothesis-generating work, further experimental and clinical validation is warranted.

Skin wound healing remains challenging due to high infection rates and inefficient repair, with traditional dressings lacking mechanical durability and antibacterial control for dynamic wound environments. Herein, an active wound dressing (AWD) was fabricated using fish skin gelatin and chitosan via synergistic covalent (MBAA-PNIPAm) and ionic (Ca2+-FSG) crosslinking, integrated with silver nanoparticles (AgNPs), and optimized by finite element modeling. Studies have demonstrated that AWD exhibits outstanding mechanical properties: a tensile strain of 600.05% ± 25.32% and a maximum fracture energy of 2000.47 ± 150.67 J m−2. It also demonstrates efficient thermal responsiveness, with a volume shrinkage rate of 22.17% ± 2.34% at 37 °C over 3 hours. Additionally, it exhibits potent antibacterial activity, achieving an antibacterial rate exceeding 89% against three pathogenic bacteria. Furthermore, it demonstrates excellent biocompatibility, with a cell survival rate exceeding 85% and no significant Ag+ accumulation. In vivo experiments showed that the wound contraction rate in mice treated with AWD reached 68.67% ± 4.56% within 8 days, significantly promoting granulation tissue formation and epithelial regeneration. The prediction error between the finite element model and experimental results was only 6.61%. This temperature-responsive AWD combines mechanical robustness, antibacterial efficacy, and wound contraction ability, expanding applications of fish-derived gelatin and chitosan. It offers a promising strategy for intelligent trauma repair materials with clinical translation potential.