Yanbian University

Dark tea, a post-fermented tea, has traditionally been used to regulate liver disorders. As an ethnomedicinal plant, its efficacy in alleviating chronic liver disease has been demonstrated.

This study explored the protective effect and potential mechanism of dark tea extract (DTE) against hepatic fibrosis.

The chemical ingredients of DTE were analyzed by UPLC-MS, and active ingredients were identified using a multidimensional biomolecular assay. BALB/c mice were induced with thioacetamide (TAA) and then treated with DTE (100, 200, or 400 mg/kg) or curcumin. Hepatic stellate cells (HSCs) were stimulated with TGF-β or conditional medium from LPS-primed liver sinusoidal endothelial cells (LSECs), and then treated with DTE (50, 100, 200 μg/mL), GW4064, and Guggulsterones, respectively.

In vivo, DTE significantly reduced serum transaminase levels and collagen deposition, and alleviated liver and intestinal histopathological damage in TAA-induced mice. DTE upregulated the expression of farnesoid X receptor (FXR) and Takeda G protein-coupled receptor 5 (TGR5), inhibited inflammatory responses, and reduced intestinal permeability by regulating Occludin and ZO-1 expressions. Additionally, DTE regulated VEGFA, CD34 and LYVE1 expressions to improve hepatic sinusoidal capillarization. In vitro, DTE reversed HSCs activation by regulating extracellular matrix (ECM) accumulation, inflammation, and FXR-TGR5 expressions. Deficiency of FXR caused by an antagonist or siRNA blocked the regulation of DTE on ECM accumulation and inflammation. DTE downregulated CD34 and upregulated LYVE1 expressions, suppressed capillarization of LSECs and restored their differentiated phenotype. DTE further reversed activated HSCs stimulated by LSECs capillarization, which was related to the activation of FXR and TGR5.

DTE improves liver fibrosis by regulating collagen deposition, inflammation, intestinal permeability, and liver sinusoidal capillarization, which is a process closely related to FXR/TGR5.

α-Dicarbonyl compounds (α-DCs) in human saliva are closely associated with diabetes, but their inherent instability presents challenges for sample transportation and detection. This study introduces a novel approach for the simultaneous detection of three α-DCs-3-deoxyglucoside (3-DG), glyoxal (GO), and methylglyoxal (MG)-in dried saliva spots using 4,5-dimethyl-1,2-phenylenediamine (DMPD) derivatization combined with UHPLC-MS/MS. 5.0 mm diameter double-circle qualitative filter paper was selected as the substrate for dried saliva, with optimal extraction conditions involving two 3-minute extractions. Chromatographic separation was performed on a CSH™ C18 column (2.1 × 50 mm, 1.7 μm) via gradient elution, utilizing a mobile phase consisting of 0.1 % formic acid in water and 0.1 % formic acid in acetonitrile. The entire separation process was completed within a 5.0 min timeframe. The linearity of 3-DG and GO in the concentration range of 0.10-12.5 μM, and MG in the range of 0.24-31.25 μM. (R² ≥ 0.9994). Intra- and inter-day precision yielded RSD values ranging from 0.59 % to 5.78 %, with average recoveries between 99.78 % and 108.50 %. The detection limits were 0.85-12.85 fmol. This method was applied to analyze dried saliva samples from 44 healthy volunteers (HVs) and 46 diabetic patients (DPs), revealing significant differences (p < 0.001) in the concentrations of 3-DG, GO, and MG between HVs and DPs. The research outcomes validate the practicality of α-DCs in dried saliva samples as prospective biomarker candidates for diabetes, while introducing an innovative methodology for their utilization in clinical settings.

Microplastic (MP) pollution challenges marine ecosystems, with gastropods like Haliotis discus hannai being potentially vulnerable. However, the reversibility of MP-induced physiological damage in gastropods remains poorly understood. This study investigated the effects of polystyrene microplastics (PS-MPs; 0, 100, and 1000 μg/L) on H. discus hannai in a 14-day exposure followed by a 7-day recovery experimental design. The analysis focused on antioxidant status, energy metabolism, and immune pathways. Results showed dose-dependent oxidative stress during PS-MPs exposure, indicated by significantly (P < 0.05) elevated antioxidant enzyme activities (Superoxide dismutase, Catalase and Glutathione peroxidase), which decreased but did not normalize during recovery. Exposure to a low concentration of PS-MPs (100 μg/L) resulted in metabolic adjustments consistent with homeostatic maintenance, whereas a high concentration (1000 μg/L) significantly increased lactate dehydrogenase and Na⁺/K⁺-ATPase activities, suggesting metabolic disruption. These energy metabolism alterations persisted, showing incomplete recovery. Immune pathway analysis revealed minimal changes at low PS-MPs concentrations but significant enrichment of pathways like IL-17 and Fc-Epsilon-RI signaling at high concentrations during exposure. Notably, sustained activation of immune pathways, including NOD-like receptor, Toll-like receptor, and IL-17 signaling (particularly in the high-concentration group), persisted during recovery, potentially indicating prolonged cellular stress or links to apoptosis. Furthermore, Integrated Biomarker Response (IBR) analysis confirmed that H. discus hannai remained severely impacted even after the recovery period. This study provides crucial evidence on the complex toxicodynamics of PS-MPs in H. discus hannai, highlighting incomplete physiological recovery and significant damage during the recovery period following exposure, especially at high concentrations.