Background: Dry eye is a common ocular surface disease characterized by corneal epithelial damage, yet effective therapies targeting mitochondrial dysfunction remain limited. Purpose: To investigate the protective effects of honokiol (HKL) against dry eye-related corneal epithelial injury and elucidate the underlying mechanisms focusing on SIRT3 and PINK1-mediated mitophagy. Methods: A benzalkonium chloride (BAC)-induced dry eye mouse model (n=6) was established and treated with HKL eye drops. Corneal damage was assessed by slit-lamp photography, Schirmer I test, and H&E staining. SIRT3, PINK1, and mitophagy-related proteins were examined by immunofluorescence and Western blot. Human corneal epithelial cells (HCECs) were exposed to hyperosmolar stress with or without HKL (n=5). Cell viability, oxidative stress, mitochondrial morphology, and function were evaluated. PINK1 knockdown was performed to validate its role in HKL-mediated protection. Results: In both BAC-induced mice and hyperosmolar-stressed HCECs, SIRT3 was downregulated with activation of PINK1/Parkin-mediated mitophagy, accompanied by mitochondrial dysfunction, apoptosis (increased to 36.2%, P < 0.001), and inflammation. HKL treatment restored mitochondrial dynamics balance, enhanced PINK1-dependent mitophagy, and alleviated oxidative stress, apoptosis (reduced to 17.4%, P < 0.001), and inflammation. PINK1 knockdown abrogated these protective effects, confirming PINK1 as a key mediator. Notably, HKL primarily enhanced SIRT3 activity rather than its expression, as immunofluorescence showed partial SIRT3 recovery in the corneal epithelium while Western blot showed no statistical significance. In vivo, HKL improved corneal integrity, tear secretion (from 1.8 to 3.5 mm/2min, P < 0.01), and tissue structure, consistent with in vitro findings. Conclusion: This study demonstrates that HKL protects against dry eye-associated corneal epithelial damage potentially through a mechanism involving SIRT3 modulation and PINK1-mediated mitophagy, offering a potential therapeutic strategy for dry eye disease.
Background: Drug-induced liver injury (DILI) presents a significant clinical challenge with limited preventive and therapeutic options. Artesunate (ART) holds potential for DILI treatment; however, its precise hepatoprotective mechanisms and direct molecular target(s) remain unclear. Methods: We evaluated ART in a podophyllotoxin (PPT)-induced rat model of DILI. Its direct target was identified through integrated chemical proteomics (pull-down/LC-MS/MS) followed by biophysical validation (SPR, MST) and molecular dynamics simulations. Mechanistic insights from hepatic transcriptomics (RNA-seq) were confirmed by Western blot, and extended with targeted metabolomics and 16S rRNA sequencing of gut microbiota. Results: ART administration conferred significant hepatoprotection in the PPT-induced rat model, as evidenced by dose-dependent amelioration of key serum markers of liver injury (P < 0.05), attenuation of hepatic oxidative stress, and improved histopathological outcomes. Mechanistically, we identified mannose-binding lectin 2 (MBL2) as a direct, high-affinity target of ART (KD approximate to 3.84 & micro;M). Upon binding to MBL2, ART suppressed the hepatic TLR4/NF-kappa B inflammatory axis and, in parallel, relieved NF-kappa B-mediated repression of NRF2, thereby cooperatively activating the NRF2-driven antioxidant defense program. Integrated multi-omics analysis further revealed that ART remodeled the gut microbiota, enriching beneficial genera including Lactobacillus, and that the abundance of these bacteria correlated positively with hepatic levels of antioxidant metabolites (P < 0.05), supporting a functional role of the gut-liver axis in its systemic protective effects. Conclusion: This study elucidates that ART alleviates DILI through targeting MBL2, thereby dually modulating the NF-kappa B/NRF2 signaling axis to simultaneously suppress inflammatory responses and enhance endogenous antioxidant defenses. These findings provide a novel and robust pharmacological basis for repositioning ART for the clinical prevention and treatment of DILI.
Background: The increasing incidence of invasive fungal infections, coupled with the rising threat of antifungal resistance and tolerance, has created an urgent demand for novel antifungal agents with distinct mechanisms of action. Methods: In this study, a series of caffeic acid amide derivatives (59 compounds) from our in-house library was systematically screened for in vitro antifungal activity against clinically relevant pathogenic fungi. Structure-activity relationship (SAR) analysis was performed to identify key pharmacophoric features. The lead compound F4 was further characterized via checkerboard microdilution assays, biofilm formation inhibition assays, and spot assays to evaluate its combination effects, antibiofilm potential, and ability to Results: SAR studies revealed that the ortho-dihydroxy moiety and thioamide substitution are critical determinants of antifungal activity. F4 demonstrated potent and broad-spectrum activity against Cryptococcus neoformans and Candida glabrata (MIC50 = 0.5-16 gg/mL), and moderate activity against drug-resistant Candida auris (MIC50 = 1-8 gg/mL), outperforming FLC against certain strains. Furthermore, F4 significantly inhibited biofilm formation in a dose-dependent manner across multiple species, with strain-dependent potency relative to FLC. Checkerboard assays indicated additive effects when F4 was combined with FLC against C. auris, C. glabrata, and C. neoformans. Importantly, although F4 showed no synergy and lacked direct growth inhibition in YPD medium (MIC >= 64 gg/mL) against Candida albicans SC5314, spot assays demonstrated that F4 at concentrations >= 16 gg/mL effectively eliminated FLC tolerance at 37 degrees C, a temperature-dependent effect not observed at 30 degrees C. Conclusion: Compound F4 emerges as a promising lead for antifungal drug development, distinguished by its broad-spectrum activity, antibiofilm properties, and unique capacity to eliminate FLC tolerance. These findings warrant further mechanistic and in vivo investigations.
Background: This study aimed to explore the ED90 of programmed intermittent epidural bolus (PIEB) volume for labor analgesia with different concentrations of ropivacaine when combining with dural puncture epidural (DPE) techniques. Methods: Ninety patients were randomized to three groups (n=30 each) to receive an initial 12-mL bolus of ropivacaine (0.075%, 0.1%, or 0.125%) with 0.3 mu g/mL sufentanil. PIEB began at 60 min after the initial bolus, with 45-min intervals; the first patient in each group received 10 mL ropivacaine-sufentanil solution, and subsequent volumes were adjusted by +/- 2 mL via sequential allocation based on effective labor analgesia (no epidural bolus intervention administered). The primary and secondary outcomes were, respectively, the ED90 of PIEB volume at different ropivacaine concentrations for effective labor analgesia, and maternal analgesia quality and neonatal outcomes. Results: The ED90 of PIEB volume for achieving effective labor analgesia was 13.2 mL (95% CI, 11.9-14.5), 11.1 mL (95% CI, 10.0-12.2), and 9.4 mL (95% CI, 8.4-10.3) in the 0.075%, 0.1%, and 0.125% groups, respectively. No statistically significant differences in maternal analgesia quality and neonatal outcomes were identified among groups. Conclusion: When the combined DPE and PIEB technique is applied for effective labor analgesia, the ED90 of PIEB volume at concentrations of 0.075%, 0.1%, and 0.125% is 13.2 mL, 11.1 mL, and 9.4 mL respectively. Since higher concentration corresponds to a higher administered dose of ropivacaine, the ED90 of PIEB volume decreases as concentration increases.
Background: Patients undergoing transfemoral transcatheter aortic valve replacement (TAVR) under general anesthesia are particularly vulnerable to hemodynamic instability. General anesthesia remains a common approach and is necessary in selected patients, although conscious sedation is increasingly used. A previous randomized trial showed greater peri-induction hemodynamic stability with cipepofol than with propofol, but whether this translates into improved clinical outcomes is uncertain. Methods: We conducted a post hoc, exploratory extended follow-up of a single-center randomized trial. Patients were randomly assigned to cipepofol or propofol. In-hospital analyses included 122 TAVR-treated participants (61 per group), whereas the 30-day and 1-year analyses included 118 participants with follow-up data in the full analysis set (59 per group). The primary endpoint was the 1year composite of all-cause mortality, stroke, acute kidney injury, myocardial infarction, and new-onset atrial fibrillation (NOAF). For this exploratory analysis, the components were grouped based on their hypothesized association with periprocedural hemodynamic instability and were also reported separately. Results: At 1 year, the primary composite endpoint occurred in 13/59 (22.0%) patients in the cipepofol group and 22/59 (37.3%) patients in the propofol group (hazard ratio [HR] 0.56; 95% confidence interval [CI] 0.28 to 1.10; P = 0.093). NOAF, analyzed separately as an exploratory secondary endpoint, occurred less frequently with cipepofol than with propofol (7/59 [11.9%] vs 17/59 [28.8%]; HR 0.39; 95% CI 0.16 to 0.95; P = 0.037). At 30 days, the composite endpoint occurred in 9/59 (15.3%) and 14/59 (23.7%) patients, respectively (HR 0.63; 95% CI 0.27 to 1.45; P = 0.277). The in-hospital composite endpoints were 10/61 (16.4%) and 16/61 Conclusion: In this exploratory extended follow-up, cipepofol did not significantly reduce the 1-year primary composite endpoint compared with propofol. The observed lower incidence of NOAF should be regarded as hypothesis-generating given the limited sample size, multiple comparisons, and intermittent rhythm surveillance. The applicability of these findings is limited to patients undergoing transfemoral TAVR under general anesthesia and does not extend to procedures performed under conscious sedation.
