多聚甲醛溶液(4% PFA)

Background The hypoxic tumor microenvironment is a key factor that promotes metabolic reprogramming and vascular mimicry (VM) in ovarian cancer (OC) patients. ESM1, a secreted protein, plays an important role in promoting proliferation and angiogenesis in OC. However, the role of ESM1 in metabolic reprogramming and VM in the hypoxic microenvironment in OC patients has not been determined. Methods Liquid chromatography coupled with tandem MS was used to analyze CAOV3 and OV90 cells. Interactions between ESM1, PKM2, UBA2, and SUMO1 were detected by GST pull-down, Co-IP, and molecular docking. The effects of the ESM1-PKM2 axis on cell glucose metabolism were analyzed based on an ECAR experiment. The biological effects of the signaling axis on OC cells were detected by tubule formation, transwell assay, RT‒PCR, Western blot, immunofluorescence, and in vivo xenograft tumor experiments. Results Our findings demonstrated that hypoxia induces the upregulation of ESM1 expression through the transcription of HIF-1α. ESM1 serves as a crucial mediator of the interaction between PKM2 and UBA2, facilitating the SUMOylation of PKM2 and the subsequent formation of PKM2 dimers. This process promotes the Warburg effect and facilitates the nuclear translocation of PKM2, ultimately leading to the phosphorylation of STAT3. These molecular events contribute to the promotion of ovarian cancer glycolysis and vasculogenic mimicry. Furthermore, our study revealed that Shikonin effectively inhibits the molecular interaction between ESM1 and PKM2, consequently preventing the formation of PKM2 dimers and thereby inhibiting ovarian cancer glycolysis, fatty acid synthesis and vasculogenic mimicry. Conclusion Our findings demonstrated that hypoxia increases ESM1 expression through the transcriptional regulation of HIF-1α to induce dimerization via PKM2 SUMOylation, which promotes the OC Warburg effect and VM.

Bioengineered tracheas have shown considerable potential in tracheal injury repair; however, their practical value is limited by challenges in tracheal cartilage regeneration, and postoperative tracheal stenosis remains a common issue. Here, inspired by the 3-layer structure of the trachea and the multi-segmental characteristics of its cartilage, a multilayered bioengineered tracheal scaffold (named Sd@d-ECM/BMSCs/SilMA) with a microgroove structure is designed in this study to repair tracheal defects. In this design, the microgrooved surface of the methacrylated silk fibroin (SilMA) hydrogel provides spatial guidance for the directional growth of bone marrow mesenchymal stem cells (BMSCs) and enhances their adhesion and proliferation. The extracellular matrix of the decellularized cartilage scaffold offers the necessary microenvironment and mechanical support for BMSCs to differentiate into cartilage. Under the influence of the dual-layer structure (inner and outer), the middle-layer BMSCs can undergo stable chondrogenic differentiation without any inducing agents. Sd@d-ECM/BMSCs/SilMA effectively promotes tracheal cartilage formation in a rabbit defect model, reduces the incidence of tracheal stenosis, and substantially improves respiratory function. Sd@d-ECM/BMSCs/SilMA not only confirms the successful construction of microgroove structures on the surface of the SilMA hydrogel and the effective loading of BMSCs but also demonstrates significant experimental value in tracheal cartilage repair and regenerative medicine.

Rationale: Spinal cord injury (SCI)-induced vascular damage causes ischemia and hypoxia at the injury site, which, in turn, leads to profound metabolic disruptions. The effects of these metabolic alterations on neural tissue remodeling and functional recovery have yet to be elucidated. The current study aimed to investigate the consequences of the SCI-induced hypoxic environment at the epicenter of the injury. Methods: This study employed metabolomics to assess changes in energy metabolism after SCI. The use of a lactate sensor identified lactate shuttle between endothelial cells (ECs) and neurons. Reanalysis of single-cell RNA sequencing data demonstrated reduced MCT1 expression in ECs after SCI. Additionally, an adeno-associated virus (AAV) overexpressing MCT1 was utilized to elucidate its role in endothelial-neuronal interactions, tissue repair, and functional recovery. Results: The findings revealed markedly decreased monocarboxylate transporter 1 (MCT1) expression that facilitates lactate delivery to neurons to support their energy metabolism in ECs post-SCI. This decreased expression of MCT1 disrupts lactate transport to neurons, resulting in a metabolic imbalance that impedes axonal regeneration. Strikingly, our results suggested that administering adeno-associated virus specifically to ECs to restore MCT1 expression enhances axonal regeneration and improves functional recovery in SCI mice. These findings indicate a novel link between lactate shuttling from endothelial cells to neurons following SCI and subsequent neural functional recovery. Conclusion: In summary, the current study highlights a novel metabolic pathway for therapeutic interventions in the treatment of SCI. Additionally, our findings indicate the potential benefits of targeting lactate transport mechanisms in recovery from SCI.

Background Hepatocellular carcinoma (HCC) is a global health challenge with high mortality rates, particularly in patients with advanced disease and lung metastasis. T-cell receptor (TCR)-T cell therapy based on specific neoantigens, is an emerging treatment with potential for HCC. However, the prognosis of patients remains poor, underscoring the need for novel targets and strategies. Methods We conducted a comprehensive study to investigate the role of C7orf50 and its neoantigens in HCC. We evaluated the functional impact on HCC progression and metastasis in vitro and in vivo , and further explored the mechanism by which C7orf50 promotes cancer metastasis and remodels tumor immune environment. Using exome and transcriptome sequencing, we identified neoantigens associated with C7orf50 and assessed their potential in TCR-T therapy. Results Our in vitro experiments revealed that C7orf50 overexpression enhances HCC cell proliferation, migration, and invasion, while knockdown inhibits these processes. In vivo , C7orf50 promoted tumor growth and lung metastasis, with a significant correlation between C7orf50 expression and poor clinical outcomes in patients with HCC. We further demonstrated that C7orf50 activates the NF-κB/PAI-1 pathway by binding to AEG-1 and facilitating its nuclear translocation, thereby promoting tumor-associated macrophage recruitment. Meanwhile, we found that TCR-T from C7orf50 -related neoantigen could obviously realize the killing effect on HCC cells, revealing its great role in cell therapy. Conclusion C7orf50 is a critical mediator of HCC progression and lung metastasis, acting through the NF-κB/PAI-1 pathway and AEG-1. Its expression levels, along with those of PAI-1 and CD68, serve as independent prognostic markers. And C7orf50 -related neoantigen shows great application potential in TCR-T therapy. These findings provide a foundation for developing C7orf50 -targeted therapies and highlight its potential in precision medicine and immunotherapy for HCC.

Hydrophilic thermoplastic polyurethane (TPU) is applicable in the fields of packaging, oil–water separation, and biomedical. Nevertheless, the process of its preparation is still a great challenge. The objective of this study is to develop TPU with excellent wettability and hydrophilic stability by surface modification. Four methods were evaluated: coating with PVP (polyvinylpyrrolidone) solution, immersion in PVP solution, N 2 /Ar plasma treatment, and N 2 /Ar plasma-induced graft polymerization with PVP. Finally, the results indicate that the sample called TPU-g-PVP, which utilizes plasma-induced grafting, exhibits the best performance. The water contact angle (WCA) reduces from 109° to 9.2°, and maintains within 20° after 80 days under atmospheric conditions. Its surface energy (72.4 mN/m) is increased by 2.8 times than untreated TPU. The surface morphology and roughness are almost unchanged when compared to the original TPU. Some characteristic peaks (–OH and C = O) of PVP on the TPU-g-PVP were found by infrared spectroscopy. X-ray photoelectron spectroscopy (XPS) examination revealed a reduction in C on the TPU surface as the content of N and O increases. Experiments on biocompatibility showed that the PVP deposited substrates improved cell adherence without generating new cytotoxicity.

Interleukin-33 (IL-33), an emerging cytokine within the IL-1 family, assumes a pivotal function in the control of obesity. However, the specific mechanism of its regulation of obesity formation remains unclear. In this study, we found that the expression level of IL-33 increased in visceral adipose tissue in mice fed with a high-fat diet (HFD) compared with that in mice fed with a normal diet (ND). In vitro, we also found the expression level of IL-33 was upregulated during the adipogenesis of 3T3-L1 cells. Functional test results showed that knockdown of IL-33 in 3T3-L1 cells differentiation could promote the accumulation of lipid droplets, the content of triglyceride and the expression of adipogenic–related genes (i.e. PPAR-γ, C/EBPα, FABP4, LPL, Adipoq and CD36). In contrast, overexpression of IL-33 inhibits adipogenic differentiation. Meanwhile, the above tests were repeated after over-differentiation of 3T3-L1 cells induced by oleic acid, and the results showed that IL-33 played a more significant role in the regulation of adipogenesis. To explore the mechanism, transcriptome sequencing was performed and results showed that IL-33 regulated the PPAR signaling pathway in 3T3-L1 cells. Further, Western blot and confocal microscopy showed that the inhibition of IL-33 could promote PPAR-γ expression by inhibiting the Wnt/β-catenin signal in 3T3-L1 cells. This study demonstrated that IL-33 was an important regulator of preadipocyte differentiation and inhibited adipogenesis by regulating the Wnt/β-catenin/PPAR-γ signaling pathway, which provided a new insight for further research on IL-33 as a new intervention target for metabolic disorders.

Background Hepatocellular carcinoma (HCC) exhibits high recurrence rates and limited therapeutic options. Endothelial cell-specific molecule 1 (ESM1) and angiopoietin-like 4 (ANGPTL4) are implicated in tumor progression, yet their synergistic role in HCC lipid metabolism and angiogenesis remains unexplored. Methods We integrated multi-omics approaches, including RNA sequencing, metabolomics, and immunoprecipitation-mass spectrometry, in HCC cell lines and patient-derived xenograft models. Key experiments involved Co-IP, Western blotting, tube formation assays, and clinical tissue microarray analysis to validate the ESM1-ANGPTL4-FASN-trioleate axis. Results ESM1 and ANGPTL4 formed a positive feedback loop, stabilizing fatty acid synthase (FASN) to promote trioleate synthesis. Trioleate activated the NF-κB/IL-17 pathway in HCC cells and upregulated CD99 in endothelial cells, driving angiogenesis. In vivo, ESM1/ANGPTL4 knockdown suppressed tumor growth, which was rescued by trioleate supplementation. Clinical data revealed elevated ESM1/ANGPTL4 expression in bevacizumab-resistant HCC, correlating with poor prognosis. Conclusions The ESM1-ANGPTL4-FASN-trioleate axis orchestrates metabolic reprogramming and endothelial activation, representing a promising therapeutic target. Future studies should explore combination therapies targeting this axis and overcoming bevacizumab resistance in HCC.

Knee osteoarthritis (KOA) is a chronic disease characterized by joint wear and cartilage degeneration. Current clinical treatments are based on symptomatic relief and are not effective in regenerating cartilage, and inflammation-induced cartilage damage accelerates the progression of osteoarthritis, making the protection of articular cartilage important for controlling the development of knee osteoarthritis. In this study, a biodegradable hydrogel (HA-Ca-Alg@Ica) loaded with Icariin (Ica) was prepared by in situ cross-linking of hyaluronic acid-calcium complex (HA-Ca) and sodium alginate (Alg-Na) for local sustained delivery of Ica. The hydrogel promoted chondrocyte proliferation and inhibited the degradation of cartilage matrix by regulating key factors (Wnt3a, β-catenin and GSK-3β) in the Wnt/β-catenin signaling pathway. In addition, the hydrogel reduced the expression of inflammatory factors, including IL-1β, IL-6, TNF-α, COX-2, and MMP13, leading to a reduction in inflammation and pain relief. In summary, this hydrogel containing Icariin has shown significant effects in reducing chondrocyte degradation and promoting chondrocyte proliferation, which can play a role in delaying osteoarthritis by protecting chondrocytes. These findings offer innovative prospects for the therapeutic management of knee osteoarthritis.

This study aimed to investigate the role of urocanic acid (UCA) modulates cognitive impairment in a D-galactose (DG)-induced aging model, providing new insights and a theoretical foundation for the treatment of related diseases. Mouse senescence and astrocyte model was created by DG-induced stimuli to assess the effects of UCA. Molecular docking was utilized to confirm potential targets of UCA, and the interaction was validated using the Drug Affinity Responsive Target Stability assay combined with Western blot analysis. The molecular mechanism of UCA was elucidated through in vivo and in vitro experiments. The result showed that UCA treatment ameliorated learning and memory capabilities in DG-induced mice, maintains astrocyte morphology while reducing apoptosis and senescent cells, and inflammatory factors. Additionally, UCA treatment alleviated cell cycle arrest. Molecular docking revealed direct binding between UCA and the ZCCHC3 protein. ZCCHC3 overexpression exacerbated cellular senescence, increased apoptosis and senescent cells, and inflammatory factor levels, while simultaneously activating the cyclic GMP-AMP synthase (cGAS)/stimulator of interferon genes (STING) pathway. UCA treatment reversed the effects of ZCCHC3 overexpression. Mechanistically, UCA inhibited the cGAS/STING pathway by binding to ZCCHC3, thereby alleviating cellular senescence. In vivo experiments further confirmed that ZCCHC3 overexpression or exogenous cGAS activation negated the cognitive protective effects of UCA. The study demonstrates that UCA alleviates cognitive impairment and astrocyte senescence by directly binding to ZCCHC3 to suppress the cGAS‑STING pathway. These results identify ZCCHC3 as a novel therapeutic target and clarify the molecular basis of related disorders.raci

Postoperative delirium (POD) is a serious complication in elderly patients undergoing surgery; however, its underlying mechanisms remain poorly understood. In this study, we investigated whether dexmedetomidine (DEX) ameliorates POD-like behavior by regulating microglial autophagy through the MAPK14 pathway. DEX targets were identified through transcriptomic analysis, and MAPK14 was validated as a key mediator by using molecular docking. In vitro experiments employed LPS-stimulated BV2 microglia to evaluate the effects of DEX on apoptosis (flow cytometry) and autophagy (transmission electron microscopy, BCL-2/Beclin-1/LC3-II/p62). In the aged C57BL/6   J mouse model of anesthesia/surgery-induced POD, animals received intraperitoneal injections of DEX or MAPK14 overexpression, followed by behavioral tests (open-field test and Y-maze test). Subsequently, hippocampal tissue was subjected to enzyme-linked immunosorbent assay and Western blot analyses to assess the levels of inflammatory factors and autophagy. DEX treatment effectively inhibited LPS-induced apoptosis in BV2 microglia while enhancing autophagy, as evidenced by increased LC3-II and Beclin-1 expression and decreased p62 and Bcl-2 levels. In the aged mouse POD model, DEX administration significantly reduced levels of neuroinflammatory factors and S100β while protecting against neuronal damage in the hippocampal CA1 and dentate gyrus regions. Behavioral assessments revealed substantial cognitive improvements in DEX-treated aged mice with delirium-like phenotype. All these therapeutic effects were abolished by MAPK14 overexpression. DEX attenuates POD by inhibiting MAPK14 to restore microglial autophagy and suppress neuroinflammation, thereby offering a novel therapeutic strategy for elderly patients. Furthermore, MAPK14 activation may serve as a biomarker for DEX responsiveness.