Liver cirrhosis represents the irreversible end-stage of various chronic liver diseases, whereas hepatic fibrosis, characterized by hepatic stellate cell (HSC) activation, constitutes the critical pathological intermediate in this progression. In this study, we investigated the antifibrotic effects of loganin, a natural iridoid glycoside, and elucidated its underlying mechanisms in bile duct ligation (BDL)-induced cholestatic liver injury and hepatic fibrosis and in TGF-β1-stimulated HSCs. Sprague-Dawley rats subjected to BDL surgery received loganin for 14 days, resulting in marked amelioration of hepatomegaly, inflammatory infiltration, hepatocyte necrosis, and collagen deposition, alongside restoration of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), and total bilirubin (T-BIL) levels and suppression of fibrogenic gene expression (collagen type I alpha 1 chain [COL1α1], alpha-smooth muscle actin [α-SMA], and fibronectin). Moreover, loganin attenuated hepatic autophagy, as evidenced by reduced Beclin-1 expression and LC3B-II/I ratio, with decreased colocalization of α-SMA and LC3B. Bioinformatics analysis and molecular docking indicated that epidermal growth factor receptor (EGFR) may be the target protein through which loganin exerts its antifibrotic effect. Experimental validation demonstrated that loganin suppressed BDL-induced EGFR phosphorylation in vivo. In TGF-β1-activated HSC-T6 cells, loganin inhibited cell activation, downregulated fibrosis markers, impaired autophagic flux, and suppressed TGF-β1-induced upregulation of Beclin-1 and LC3B-II/I. Critically, cotreatment with the EGFR agonist EGF reversed the autophagy-inhibitory and antifibrotic effects of loganin, whereas the EGFR inhibitor AG-1478 mimicked loganin's actions, confirming EGFR signaling as the functional mediator. Overall, these results indicated that loganin could alleviate liver injury and hepatic fibrosis in BDL rats by inhibiting autophagy and activation of HSCs, and this protective effect is mediated by the EGFR pathway.

Tuberculosis, caused by Mycobacterium tuberculosis (MTB), affects approximately 25% of people globally as latent infection (LTBI). Although macrophage CREB activation promotes MTB survival, the underlying mechanisms remain unclear. This study reveals, for the first time, how MTB modulates M2 macrophage polarization through the CREB1/TREM2 signaling pathway. Mononuclear macrophages were isolated from clinical sample. Flow cytometry was used to determine the M2 polarization ratio. The expression of TNF-α, IL-10, and IL-1β was determined by ELISA. The mRNA expression of iNOS, IL-1β, CD206, Arg-1, IL-10, CREB1, and TREM2 was assessed by qPCR. The protein expression of CREB1, CD206, Arg-1, IL-10, and TREM2 was evaluated by western blot. The colony-forming unit (CFU) assay was used to detect the survival of MTB. CHIP and Dual-luciferase reporter assays were used to confirm the binding of CREB1 and TREM2. Clinical sample analysis revealed that the expression levels of CREB1 and TREM2 in peripheral blood mononuclear macrophages (PBMCs) were significantly upregulated in tuberculosis patients. Following H37Rv infection, an increase in the M2 macrophage proportion was observed. Infection with MTB also elevated the protein level of IL-10, as well as CREB1 mRNA and protein expression. Transfecting sh-CREB1 into macrophages or adding CREB inhibitors 666-15 markedly reduced intracellular MTB CFU counts, implying a potential restrictive effect on viable MTB load within macrophages. Knockdown of TREM2 similarly decreased MTB CFU burden and restrained macrophage M2 polarization. Furthermore, transfecting sh-CREB1 into macrophages can suppress the proliferation of MTB within macrophages, and these effects could be further counteracted by the action of oe-TREM2. These results elucidated that MTB promoted M2 macrophage polarization through CREB1/TREM2.

This study evaluated both the acute and sublethal effects of pyriproxyfen in Cyprinus carpio using an integrated multibiomarker approach. The 96-h LC50 value was calculated as 4.525 mg/L. Following 14 days of sublethal exposure, pyriproxyfen caused clear, dose-dependent changes across hematological, biochemical, genotoxic, and molecular parameters. Hemoglobin and hematocrit levels declined progressively with increasing concentration, pointing to the development of anemia-like conditions. Biochemical analyses revealed a biphasic response pattern: Several parameters initially increased at lower concentrations but declined at higher doses, suggesting a shift from adaptive metabolic responses to hepatic impairment. DNA damage, assessed by the comet assay, increased significantly in all exposed groups, indicating genotoxic effects even at relatively low concentrations. At the molecular level, antioxidant and proinflammatory genes were upregulated at low to moderate doses, reflecting an early defense response. However, their expression was suppressed at higher concentrations, implying exhaustion of protective mechanisms. In contrast, HSP70 expression increased steadily with dose, highlighting the intensification of cellular stress and protein damage. Overall, the findings demonstrate that pyriproxyfen exposure triggers an initial adaptive response that progressively transitions into physiological dysfunction at higher concentrations, underscoring its potential ecological risk to aquatic organisms.

Lecithin-cholesterol acyltransferase (LCAT), the key enzyme in reverse cholesterol transport (RCT), has been traditionally recognized for playing roles in high-density lipoprotein (HDL) maturation and systemic cholesterol homeostasis. However, its function in liver disease remains elusive. Here, we identify the role of LCAT in driving hepatic steatosis. Using a hepatocyte-specific lcat-overexpressing zebrafish model combined with transcriptomic and metabolic phenotyping, we demonstrated that lcat overexpression induces triglyceride (TG) accumulation and cytoplasmic lipid droplet deposition in the liver. Unexpectedly, transcriptomic profiling revealed an upregulation of fatty acid elongation pathway genes in lcat-overexpressed hepatocytes. Crucially, genetic knockdown or pharmacological inhibition of fatty acid elongation enzymes (hsd17b12a and Elovl1) alleviated LCAT-induced hepatic steatosis. Furthermore, overexpression of hsd17b12a and elovl1 resulted in hepatic steatosis similar to that of lcat-overexpressing larvae. These findings establish a novel role for LCAT in hepatic steatosis through regulating fatty acid metabolism, providing a mechanistic basis for targeting Lcat/fatty acid elongation axis in metabolic dysfunction-associated steatotic liver disease (MASLD).

Nail bed defects caused by trauma or disease frequently result in nail plate loss, persistent pain, impaired hand function, and limited regenerative healing due to the highly specialized architecture of the nail bed. However, current treatment options are inadequate because they fail to simultaneously provide facile application, robust tissue adhesion, and regenerative support. Herein, we report KeratinGel, an injectable, biodegradable, and biocompatible human hair keratin-based tissue adhesive developed for nail bed repair. KeratinGel was engineered using a dual-cross-linking strategy that combined thiol-Michael addition and visible-light-induced photocross-linking, enabling rapid in situ gelation and stable integration into the tissue. To address the brittleness of keratin-based networks while maintaining cytocompatibility, reduced keratin (Keratin-SH), chosen for its biomimetic and bioactive properties, was incorporated into methacrylated gelatin (GelMA). The KeratinGel hydrogels were evaluated as tissue adhesives and sealants through lap shear, wound closure, and burst pressure tests. The adhesive and regenerative properties of KeratinGel were assessed using a novel three-dimensional (3D) in vitro nail bed wound model that mimics the layered cellular organization of the native tissue. Among the tested formulations, 5% KeratinGel exhibited the highest cytocompatibility, achieving approximately 80% cell viability. SEM and histological analyses confirmed a homogeneous cellular distribution and preservation of structural integrity. Ex vivo testing demonstrated formulation-dependent sealing performance, while keratin incorporation preserved comparable lap shear and wound closure strengths across the formulations. Gene expression analysis revealed the upregulation of MMP-3 and MMP-19, while p16 levels remained comparable to those in the control, indicating the absence of senescence-associated damage. Enhanced PCNA and KRT14 expression further underscored the regenerative potential of KeratinGel in nail-bed-tissue engineering.

Adipose tissue senescence is increasingly recognized as a key driver of systemic aging and age-related functional decline, yet the endocrine regulators that actively promote this process remain poorly defined. Angiopoietin-like protein 8 (ANGPTL8) is a metabolic factor implicated in lipid metabolism and inflammation and has been associated with multiple aging-related disorders. However, its direct role in adipose tissue senescence and organismal aging remains unclear. Here, we identify ANGPTL8 as a previously unrecognized regulator of adipose tissue aging through integrative analyzes of human cohorts, animal models, transcriptomics, and cellular studies. In a large human cohort, circulating ANGPTL8 levels were strongly associated with biological aging and mortality risk and significantly improved machine learning-based models for age and survival prediction. Consistent with these findings, genetic deletion of Angptl8 in mice extended lifespan, attenuated aging-associated functional decline, and reduced senescence markers in adipose tissue. Transcriptomic analyzes revealed age-dependent upregulation of ANGPTL8 in adipocytes accompanied by activation of pro-senescent transcriptional programs. Mechanistically, ANGPTL8 directly interacted with AKT2 and activated the AKT-mTOR-S6K signaling pathway, thereby promoting cell-autonomous adipocyte senescence. Genetic or pharmacological inhibition of this pathway abolished the pro-senescent effects of ANGPTL8. Collectively, our findings identify ANGPTL8 as an endocrine regulator linking metabolic dysfunction to adipose tissue senescence and systemic aging, highlighting the ANGPTL8-AKT2-mTOR axis as a potential therapeutic target for delaying age-associated functional decline.

Currently, the repair of bone defects faces multiple challenges, including insufficient angiogenesis and limited osteogenic differentiation capacity, which pose significant obstacles to treatment. In this study, a ternary composite scaffold composed of PCL-nHA-nMgO (PHM) was fabricated using 3D printing technology. After surface modification with a photothermal polydopamine (PDA) coating, a PHM@PDA composite scaffold was obtained. Metformin (Met) and PDA were loaded into GelMA hydrogels to produce GPM hydrogels. The GPM hydrogels were then packed into the PHM@PDA composite scaffolds, resulting in near-infrared region-I (NIR-I)-responsive PHM@PDA/GPM composite scaffolds. The outer GPM layer serves as a three-dimensional in vivo-like microenvironment, providing physical transport channels for cellular nutrient exchange. Upon stimulation by NIR-I, the scaffold enables the dynamic release of Mg2+ and Met, upregulates the expression of the VEGF and KDR, accelerates vascular network reconstruction, and creates a mild microenvironment conducive to cell proliferation. Furthermore, by activating the PI3K/AKT signaling pathway, it promotes the expression of osteogenesis-related factors (such as transcription factor Sp7, alkaline phosphatase, and osteocalcin), thereby accelerating the osteogenic differentiation process. In vivo animal experiments, micro-CT and tissue staining confirmed that, under near-infrared light (NIR-I) stimulation, this scaffold significantly increased bone mineral density, trabecular bone thickness, and trabecular bone count and demonstrated excellent angiogenic effects. In summary, this NIR-I-responsive composite scaffold can meet the dual requirements of reconstructing the vascular network and promoting osteogenic differentiation, enabling personalized precision therapy and offering a promising new treatment strategy for complex bone defects where vascular regeneration and osteogenic differentiation are challenging.

ClpX functions as a component of the ClpXP protease, a conserved intracellular protease that regulates protein turnover, stress responses, and virulence in multiple bacterial species. Our lab has established that clpX is necessary for resistance to cell envelope targeting antibiotics, such as penicillin and daptomycin in Bacillus anthracis Sterne. Previous microarray data identified the msrA/B gene encoding a bifunctional methionine sulfoxide reductase as upregulated in the ΔclpX mutant. Methionine sulfoxide reductases (Msr) repair oxidatively damaged proteins by reducing methionine sulfoxide residues back to methionine. While Msr enzymes are primarily associated with oxidative stress, cell wall antibiotics induce expression of msrA1 and msrB in S. aureus. Here, we investigated the role of MsrA/B in oxidative and cell envelope stress. Our results show that although hydrogen peroxide and paraquat induce msrA/B expression, the ΔmsrA/B strain was not susceptible to either oxidant, whereas the ΔclpX strain was sensitive to both. We also found that loss of msrA/B conferred penicillin-specific sensitivity, but, unlike ΔclpX, increased sensitivity was not seen with other cell wall or cell membrane targeting antibiotics. Inactivation of the catalytic cysteine of either Msr domain of MsrA/B failed to complement, suggesting that the reducing activity of MsrA/B is required for penicillin resistance. These findings indicate that while MsrA/B contributes to penicillin resistance, other proteins in the ClpXP modulon must also play a role in oxidative and cell envelope stress.

Toxoplasma gondii infection poses potential risks to stem cell-based therapeutic applications; however, the effects of parasite-derived soluble factors on mesenchymal stem cells remain unclear. Here, we investigated the impact of T. gondii-derived soluble factors (Tg-SFs) on human umbilical cord-derived mesenchymal stem cells (hUC-MSCs). hUC-MSCs were exposed to Tg-SFs generated in a Transwell co-culture system, and cellular phenotypes and p53 signalling were analysed using integrated functional and molecular approaches. Tg-SFs induced upper-chamber multiplicity of infection (MOI)-dependent cytotoxicity in hUC-MSCs, accompanied by cytoskeletal remodelling and nuclear condensation. Tg-SFs exposure caused marked G2/M phase accumulation, which was accompanied by reduced expression of Cyclin A2, Cyclin B1, and Cyclin E2 and increased p21 expression. In addition, Tg-SFs exposure reduced phosphorylation of CDK1 at Thr161. Tg-SFs-exposed hUC-MSCs underwent significant apoptosis, as evidenced by PARP and Caspase-3 cleavage. Mechanistically, Tg-SFs promoted accumulation of total p53 protein and selective phosphorylation of p53 at Ser15 and Ser392, and modulated Bcl-2 family proteins by upregulating Bax, BID, Bak, Bad, and Puma while downregulating Bcl-xL and Mcl-1. Pharmacological inhibition of p53 with PFT-α and siRNA-mediated p53 knockdown partially reversed these effects, indicating that p53 signalling contributes substantially, but not exclusively, to Tg-SFs-induced mitochondrial apoptosis. These findings indicate that T. gondii-derived soluble factors impair hUC-MSC viability through G2/M phase accumulation and p53-associated mitochondrial apoptosis, highlighting a potential safety consideration for MSC-based applications under parasite-associated conditions.