DAPI染色液(10μg/mL)

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.

Although tea consumption has been suggested to affect kidney stone formation, epidemiological evidence remains inconsistent, and the underlying molecular mechanisms are unclear. To assess the association between tea intake and kidney stone risk, we initially conducted a prospective cohort analysis of 481,393 participants from the UK Biobank and a 2-sample Mendelian randomization (MR) analysis. Our findings revealed that heavy tea drinkers (>5 cups/day) had a significantly reduced risk of kidney stones (hazard ratio: 0.79, 95% confidence interval [CI]: 0.72 to 0.86, P < 0.001), and MR analyses confirmed a causal association (inverse variance weighted OR: 0.45, 95% CI: 0.32 to 0.62, P < 0.001). We next explored the effect of epigallocatechin gallate (EGCG), the main bioactive component in tea, on calcium oxalate (CaOx) stone formation. EGCG was found to inhibit the glucose-regulated protein 94/phosphatidylinositol 3-kinase/protein kinase B (GRP94/PI3K/AKT) pathway in human proximal renal tubular epithelial cells, thereby attenuating CaOx crystal-induced oxidative stress and inflammation, and inhibiting crystal-cell adhesion. This finding aligned with the observation that the activated GRP94/PI3K/AKT pathway was positively associated with inflammation-related molecules in renal papillary tissues of CaOx stone formers. Moreover, to enhance renal targeting and therapeutic potential, we synthesized cell membrane-coated EGCG-loaded poly(lactic-co-glycolic acid) (TP-EGCG) nanoparticles, which enhanced renal EGCG delivery and substantially reduced CaOx crystal deposition in a mouse model of CaOx nephrolithiasis. In conclusion, tea consumption protects against kidney stone formation, an effect exerted by EGCG through the GRP94/PI3K/AKT axis, and our novel TP-EGCG nanoparticles show strong potential for targeted prevention and treatment.

Chemodynamic therapy (CDT) is a highly promising cancer treatment strategy. However, its clinical application is severely limited due to insufficient H 2 O 2 levels. To overcome this critical issues, based on self-supply H 2 O 2 strategy this study designed a multifunctional nano-reactor hyaluronic acid (HA)-cinnamaldehyde( CA )Schiff base(HA-CA)@hollow CaO 2 (HMCaO 2 )/Copper doped luteolin carbon dots (CuLCDs) (abbreviation:HACOCLC). HACOCLC based on self-supplied H 2 O 2 /O 2 strategy to enhance the anti-tumor effect. HACOCLC preferentially enrich at the tumor site under the tumor targeting effect of HA. pH stimulation triggered the release of HMCaO 2 and CuLCDs from HACOCLC. HMCaO 2 hydrolyzed to produce H 2 O 2 , Ca 2+ and O 2 , H 2 O 2 compensated substrates for CDT, O 2 down-regulates hypoxia-inducing factor (HIF-1α) and programmed death ligand 1(PD-L1) to consolidate the therapeutic effect, and Ca 2+ entered mitochondria to cause calcium overload and induced cell death. CuLCDs underwent Fenton-like reaction to produce •OH (CDT effect) and O 2 , which further increased the O 2 level. CuLCDs has near-infrared light (808 nm) thermal conversion (conversion efficiency was 40.3 %) for photothermal therapy (PTT), Cu 2+ depletion of glutathione amplified the level of reactive oxygen species (ROS) and improved the therapeutic effect. HACOCLC self-supplied H 2 O 2 and O 2 to enhance CDT, and effectively inhibited tumor growth through the combined action of CDT/PTT/ calcium overload.

Background Pulmonary fibrosis (PF) is a progressive interstitial lung disease marked by extracellular matrix accumulation and epithelial damage, with limited therapeutic options. Alveolar epithelial cell apoptosis is a key pathological hallmark of PF, but the upstream regulators driving this process remain unclear. Caspase-9, a central initiator of the intrinsic apoptotic pathway, has been implicated in fibrotic diseases across multiple organs. However, its role in lung fibrosis and its molecular interactions are not fully elucidated. Methods Caspase-9 expression was analyzed in human PF lung tissues, bleomycin (BLM)-induced mouse models, and TGF-β1-treated MLE-12 alveolar epithelial cells. Functional studies included pharmacological inhibition, siRNA knockdown, and overexpression of Caspase-9. Fibrosis and apoptosis were assessed using Western blot, qPCR, immunohistochemistry, TUNEL, and electron microscopy. Interaction with β-catenin was examined via co-localization, modulation, and rescue experiments. Results Caspase-9 and cleaved-Caspase-9 were significantly upregulated in fibrotic lungs and TGF-β1-stimulated epithelial cells. Caspase-9 inhibition reduced collagen deposition, improved lung architecture, and suppressed pro-fibrotic markers in mice. In MLE-12 cells, Caspase-9 knockdown attenuated TGF-β1-induced apoptosis, restored E-cadherin, and downregulated fibrotic genes. Conversely, Caspase-9 overexpression aggravated fibrosis and apoptosis. Mechanistically, Caspase-9 interacted with β-catenin, enhanced its nuclear accumulation, and promoted downstream fibrotic signaling. β-catenin silencing reversed Caspase-9-induced fibrosis, while β-catenin activation nullified the protective effects of Caspase-9 inhibition both in vitro and in vivo. These results identify a functional Caspase-9/β-catenin axis in PF progression. Conclusions Caspase-9 drives pulmonary fibrosis by promoting epithelial apoptosis and activating β-catenin signaling. Targeting the Caspase-9/β-catenin axis may offer a promising therapeutic strategy for PF. Supplementary Information The online version contains supplementary material available at 10.1186/s12967-025-07020-1.

Objective. Here, we aimed to explore the main mechanism of Yaobishu (YBS) in lumbar disc herniation (LDH). Methods and Results. Eighteen compounds that might act on LDH were obtained through a combination of network pharmacology prediction and identification by high-performance liquid chromatography-mass spectrometry. The key compounds were palmitic acid and trans-4-hydroxy-3-methoxycinnamate (cinnamate). KEGG analysis demonstrated that palmitic acid target genes mainly regulate the PPAR signaling pathway, Ras signaling pathway, and fatty acid metabolism. Cinnamate target genes were primarily involved in chemical carcinogenesis-receptor activation, lipid and atherosclerosis, the HIF-1 signaling pathway, and nitrogen metabolism. The rat LDH model was constructed using autologous nucleus pulposus tissue implantation. Differential expression gene (DEGs) related to metabolism (CDKN1A and UHRF1), inflammation (S100A9 and SOCS3), autophagy (DCN and LEPR), and apoptosis (CTSW and BCL2A1) in dorsal root ganglion (DRG) tissues of the control and LDH groups was evaluated by RNA-Seq. TNF-α stimulated DRG neuronal cells were used to establish an in vitro LDH model. YBS, palmitic acid, and cinnamate reduced the expression of substance P, CGRP, S100A9, CTSW, and cleaved caspase-3, while enhancing the expression of CDKN1A, UHRF1, PCNA, Ki67, SOCS3, DCN, LEPR, and BCL2A1, as well as telomerase activity. Pearson’s correlation analysis confirmed that DCN was positively correlated with BCL2A1, indicating that autophagy might be negatively correlated with apoptosis in LDH. YBS, palmitic acid, and cinnamate reduced the Siegal neurological score and serum IL-1β and IL-18 levels, while increasing changes in the hind paw mechanical withdrawal threshold. The RNA-Seq results further showed that YBS downregulated S100A9 and CTSW expression, while upregulating SOCS3, CDKN1A, UHRF1, DCN, LEPR, and BCL2A1 expression. Conclusion. YBS and its compounds, palmitic acid, and cinnamate, attenuated LDH by regulating the inflammatory, metabolic, autophagic, and apoptotic pathways. Our results might improve the theoretical and experimental basis for clinical applications of LDH disease treatment.

Traumatic brain injury (TBI) is a severe condition with a high mortality rate, affecting multiple organs, including the gastrointestinal (GI) tract. Ghrelin is a brain-gut peptide that regulates the microbiota-brain-gut axis, facilitating communication between the GI tract and the central nervous system. This study aimed to investigate the role of ferulic acid (FA) in regulating Ghrelin to improve TBI and GI disorders (GID) induced by controlled cortical impact (CCI). This study used CCI as the in vivo TBI model and scratch-induced injury of primary astrocytes as the in vitro TBI model. The role and mechanism of FA modulation of Ghrelin in ameliorating TBI and GID were explored using multi-omics and network pharmacology analyses. In vivo, results revealed that FA is the main active component of the Guanxin II compound and mimics its function. Significant improvement in GI hypomotility and brain injury was observed in the FA group compared to the CCI group. Concurrently, FA ameliorated intestinal barrier impairment triggered by CCI-induced reduction in the expression of Ghrelin and reduces the inflammatory response. Furthermore, 16S rRNA results indicated that CCI-induced TBI worsened gut microflora imbalance via the brain-gut axis, while gut dysbiosis aggravated brain injury. FA improved the dysbiosis of Bacteroidetes and Odoribacter mainly by targeting the Ghrelin-mediated inflammatory response. RNA-seq and network pharmacology analyses revealed that FA mainly affects inflammation-mediated pyroptosis pathways in the brain-gut axis. Additionally, experimental evidence demonstrated that FA reversed CCI-induced pyroptosis in rats and scratch injury-induced pyroptosis in astrocytes by promoting the binding of Ghrelin to GHSR, which suppressed the TLR4/NF-κB/NLRP3 pathway. Conclusively, FA could alleviate TBI and GID by promoting Ghrelin to regulate the microbiota-brain-gut axis inflammation via the Ghrelin/TLR4/NLRP3 pathway.

Background Pulmonary fibrosis (PF) is a progressive and difficult-to-heal lung disease that poses a significant threat to human life and health. This study aimed to investigate the potential pathological mechanisms of PF and to identify new avenues for the treatment of PF.Methods Clinical samples were collected to assess the effect of disulfide-bond A oxidoreductase-like protein (DsbA-L) on PF. TGF-β1-induced MLE-12 cell model and bleomycin (BLM)-induced mice model were established. Changes in physiological morphology and fibrosis were observed in the lung tissues. The degree of apoptosis and the mitochondrial function was analyzed. The expression of relative cytokines was examined. The CD68+/CD206+ ratio was determined to indicate M2 macrophage polarization.Results The expression of DsbA-L was upregulated in patients with PF and PF-like models. In vitro, DsbA-L overexpression exacerbated TGF-β1-induced the deposition of extracellular matrix (ECM), apoptosis, inflammation, and mitochondrial damage, whereas DsbA-L silencing exerted the opposite effects. DsbA-L silencing inhibited the activation of AKT1, NLRP3, and SMAD3 by TGF-β1. MLE-12 cells silencing DsbA-L limited the polarization of RAW264.7 cells towards the M2 phenotype. AKT1 agonist or NLRP3 agonist reversed the role of DsbA-L silencing in inhibiting the TGF-β1/SMAD3 pathway and M2 macrophage polarization. In vivo, DsbA-L knockout protected mice from PF-like pathological damage caused by BLM.Conclusion DsbA-L exhibited a significant profibrotic effect in lung epithelial cells and mice, which increased the levels of AKT1 and NLRP3 to activate the TGF-β1/SMAD3 pathway and M2 macrophage polarization. These findings could shed light on new clues for comprehension and treatment of PF.

Carbon dots as drug carriers have received increasing attention in nanotherapy. However, the use of chemical small molecules or polymers as precursors to carbon dots often leads to toxicity, thus limiting their practical application. In contrast, herbs are biocompatible plants with complex active ingredients, making them attractive candidates for carbon dot precursors. In this paper, we prepared antitumor carbon dots (LuCDs) using a hydrothermal method and luteolin extract as a precursor. Furthermore, by modifying copper ions on the surface of LuCDs, we obtained Cu-LuCDs. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) analysis results showed that both LuCDs and Cu-LuCDs effectively inhibited the proliferation of CT26 colon cancer cells. Additionally, these modified carbon dots exhibited photothermal conversion properties not present in the precursor alone. Under 808 nm optical excitation, they achieved photothermal conversion powers of 55 and 56%, respectively. The Cu-LuCDs effectively decompose hydrogen peroxide, generating cytotoxic •OH and oxygen through Fenton-like reactions, thereby alleviating tumor hypoxia and inhibiting the expression of hypoxia-inducing factor HIF-1α. Immunofluorescence analysis confirmed that both LuCDs and Cu-LuCDs induced immune cell death and activated systemic immune activity. When combined with the immune adjuvant αPD-L1, Cu-LuCDs successfully ablated primary tumors and significantly inhibited the growth of metastatic tumors (distal tumors). The design and analysis of the antitumor activity of Cu-LuCDs provide important inspiration for the application of herbal carbon dots.

Endometrial fibrosis is the main feature of intrauterine adhesion (IUA). The m6A methylation is involved in the process of fibrosis. However, the regulatory pathways involved in m6A methylation in endometrial fibrosis remain unclear. ALKBH5 is differentially low expressed in the endometrial tissues of IUA. Overexpression of ALKBH5 inhibits TGF-β1-induced fibrosis. In the in vivo experiment, ALKBH5 overexpression reduces the degree of endometrial fibrosis in rats. ALKBH5 regulates the m6A methylation level of FABP4 mRNA. FABP4 expression is inhibited by WT- ALKBH5 , but not by catalytically inactive MUT- ALKBH5 (H204A). The m6A reader IGF2BP2 targets FABP4 and affects FABP4 mRNA stability. Inhibition of FABP4 decreases the expression of fibrosis-related markers (α-SMA, collagen I, collagen III, and Fibronectin). In addition, serum lipid metabolism is disordered in IUA rats, and ALKBH5 overexpression could partially reverse the levels of differential lipid metabolites. In conclusion, ALKBH5 is differentially low expressed in IUA. ALKBH5 signaling regulates endometrial fibrosis through FABP4 mRNA m6A methylation and lipid metabolism. This finding can provide theory support for the potential treatment strategy development of IUA.

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.