The development of foam cells is crucial in the advancement of atherosclerosis (AS). Tanshinone IIA (Tan IIA), the primary lipophilic component of Salvia miltiorrhiza, has various pharmacological effects. Despite this, the precise role of Tan IIA in AS has not been fully elucidated. In this research, we employed ApoE-/- mice to establish an AS model. Oil Red O and HE staining indicated that Tan IIA obviously reduced plaque areas in both the aorta and aortic arch. Additionally, serum analysis revealed that Tan IIA notably decreased lipid and inflammatory factor levels in AS mice. In vitro studies showed that Tan IIA primarily prevented foam cell formation by enhancing cholesterol efflux rather than increasing lipid uptake. Mechanistic investigations reveal that Tan IIA suppresses the Hedgehog (Hh) signaling pathway by downregulating the expression of transient receptor potential melastatin 2 (TRPM2) and subsequently inhibiting Ca²⁺ influx. This cascade ultimately attenuates foam cell formation and impedes the progression of AS. Overall, this research provides a solid theoretical foundation for the potential application of Tan IIA in the treatment of AS.

Objective: To investigate the mechanism through which timosaponin AⅢ (TAⅢ)-based liposomes loaded with auranofin (AUF) (T-AUF-LPs) induce ferroptosis in anaplastic thyroid carcinoma cells. Methods: T-AUF-LPs were prepared using the thin-film hydration method, and their physicochemical properties and stability were characterized. Additionally, conventional liposomes (C-AUF-Lips) were prepared using cholesterol as the membrane material. Cellular uptake efficiency was evaluated in CAL-62 cells using coumarin-6-labeled liposomes. Cell viability was assessed via CCK-8 assay. The role of ferroptosis was confirmed using the inhibitor Ferrostatin-1 (Fer-1) and flow cytometric analysis of cell death. The expression of ferroptosis-related markers (ACSL4, NCOA4, GPX4) were detected using RT-qPCR and Western blot. Levels of intracellular iron, glutathione (GSH), and lipid peroxidation were measured. A nude mouse tumor xenograft model was established to evaluate the in vivo antitumor efficacy of T-AUF-LPs, and hematoxylin-eosin (HE) staining was performed to assess potential systemic toxicity. Results: T-AUF-LPs appeared as uniformly distributed spherical particles with an average size of (119.40±3.11) nm, which was significantly smaller than that of the conventional liposomes C-AUF-LPs (P＜0.001). The zeta potential of T-AUF-LPs was (-12.07±0.65) mV, lower than that of C-AUF-LPs (P=0.002). Furthermore, CAL-62 cells exhibited significantly enhanced cellular uptake of T-AUF-LPs compared to C-AUF-LPs. Cell experiments demonstrated that among the established groups, i.e., Control, TAⅢ, AUF, AUF+TAⅢ, C-AUF-LPs, and T-AUF-LPs, the T-AUF-LPs group exhibited the lowest half-maximal inhibitory concentration (IC50=0.66 μmol/L) and the highest CAL-62 cell mortality (14.7±1.3)%. Furthermore, this group showed significantly elevated lipid peroxidation (2.07±0.28), increased iron content (4.64±0.17), and markedly reduced glutathione (GSH) levels (0.11±0.05). At the molecular level, mRNA expression of ACSL4 and NCOA4 was up-regulated (13.10±0.94 and 7.52±0.49, respectively), while GPX4 mRNA was down-regulated (0.16±0.21). Consistently, ACSL4 and NCOA4 protein expression was increased (1.30±0.06 and 1.13±0.31, respectively), whereas GPX4 protein expression was decreased (0.31±0.18).Notably, pretreatment with the ferroptosis inhibitor Fer-1 significantly reversed T-AUF-LPs-induced cell death, reducing mortality to (8.8±0.8)%. Animal studies demonstrated that among all groups, tumor growth was most significantly suppressed in the T-AUF-LPs group, which exhibited the smallest tumor volume on day 15 post-inoculation. No significant body weight loss was observed in nude mice, and histopathological assessment of the heart, liver, lungs, and kidneys revealed no apparent toxic damage. In tumor tissues of the T-AUF-LPs group, mRNA expression of ACSL4 and NCOA4 was significantly up-regulated (1.59±0.29 and 8.65±3.48, respectively), while GPX4 mRNA expression was markedly down-regulated (0.11±0.01). Consistently, ACSL4 and NCOA4 protein levels were increased (1.26±0.31 and 1.14±0.39, respectively), whereas GPX4 protein expression was significantly reduced (0.56±0.12). Conclusion: T-AUF-LPs inhibited the growth of anaplastic thyroid carcinoma cells by activating the ferroptosis pathway.

A substantial body of evidence exists demonstrating that exposure to per- and polyfluoroalkyl substances (PFASs) poses a risk to human health. Data from epidemiological studies of those exposed occupationally or environmentally demonstrate adverse health risks, and these health effects are concordant with data from toxicological studies. Systematic reviews, conducted by agencies, organizations, and independent scientists that synthesize and integrate these data streams, have concluded that a range of health risks arise from PFAS exposure, including different types of cancer, especially kidney and testicular cancer, metabolic alterations such as increased liver enzymes and increased cholesterol, immune dysfunction such as reduced vaccination efficiency, reproductive and developmental outcomes such as low birth weight and reduced duration of breast feeding, and forms of endocrine disruption. Despite this, myths and misinformation surrounding these health risks slow efforts to protect public health from the hazards of PFAS exposure. This work addresses the most predominant of these myths and counters them with accumulated evidence from epidemiological and toxicological studies, demonstrating that exposure to PFASs poses a risk to human health.

Conventional whole-cell biosensors function as "one analyte, one sensor" systems to ensure high specificity relying on transcriptional regulatory protein cascades to induce reporter gene expression, leading to the development of distinct sensor systems to monitor multiple target substances. To address this constraint and broaden analyte detection capabilities, we developed a modular biosensing platform that couples a key metabolite-responsive biosensor with various substrate-specific enzymes to detect a range of target molecules. This study demonstrated that the integration of a general hydrogen peroxide (H2O2)-responsive biosensor with glucose oxidase, uricase, and cholesterol oxidase enabled the detection of three clinically relevant biomarkers: glucose, uric acid, and cholesterol. Upon substrate-specific enzyme conversion, these three biomarkers generate H2O2, which triggers the expression of the luxCDABE reporter gene cluster and produces bioluminescence in the H2O2-responsive biosensor Pseudomonas putida KT2440[pPahpc]. The system has demonstrated sensitive detection of glucose (10-200 µM), uric acid (5-125 µM), and cholesterol (1.25-100 µM). Validation with 17 clinical urine specimens confirmed the reliability of our system for quantifying glucose and uric acid, thereby establishing its potential diagnostic utility.IMPORTANCEThe increasing public focus on health management has fueled the development of decentralized diagnostic testing. While biosensors are ideally suited to this application area, their dependence on molecular specificity mandates individualized transducers for each target. Consequently, multiplied engineering efforts and resource expenditure substantially restrict practical deployment. This study addresses this challenge through the integration of a universal signal transduction pathway with multiple substrate-specific oxidases, enabling simultaneous quantification of diverse molecular targets within a single biosensing platform. Clinical validation with 17 urine specimens confirmed the system's robust analytical performance. Our findings establish a technological foundation for cost-effective, rapid, and multiplexed home-testing devices, showing substantial promise for advancing disease surveillance and personalized healthcare management.

Clostridioides difficile is a leading cause of healthcare-associated infections and remains an urgent global public health concern. Current FDA-approved therapies for C. difficile infection (CDI) are limited and are frequently associated with treatment failure and high rates of disease recurrence. To address this unmet clinical need, we employed a drug repurposing strategy focused on FDA-approved compounds with established safety profiles and limited systemic absorption characteristics, which are desirable for localized gastrointestinal infections. Through this approach, we identified the antifungal azoles miconazole, econazole, and tioconazole as potent inhibitors of C. difficile. These compounds exhibited robust in vitro activity against a diverse panel of clinical C. difficile isolates, with MIC50 values of 1, 2, and 2 µg/mL, respectively. Time-kill analyses demonstrated that all tested azoles exerted rapid bactericidal activity within 2 h, exceeding the killing kinetics of the current standard-of-care agents vancomycin and fidaxomicin, likely attributable to their membrane-disruptive mechanism that compromises cytoplasmic membrane integrity and leads to rapid cellular collapse. Additionally, the azoles displayed prolonged post-antibiotic effects (>12 h). Furthermore, the azoles exhibited limited activity against two representative members of the commensal intestinal microbiota (Bifidobacterium and Lactobacillus spp.) and retained antibacterial efficacy under physiologically relevant conditions, including high inoculum, variable pH, and simulated gastrointestinal fluids. Mechanistic studies supported membrane targeting as the primary mode of action. Azole exposure significantly disrupted bacterial membrane integrity, and scanning electron microscopy revealed marked morphological damage, including membrane distortion and cellular collapse, consistent with structural membrane damage. Notably, cholesterol supplementation increased C. difficile tolerance to azoles and other membrane-disrupting agents, suggesting that host cholesterol levels or dietary factors may influence CDI outcomes. Finally, in a Caenorhabditis elegans CDI infection model, all tested azoles significantly improved host survival following lethal C. difficile challenge. Collectively, our findings highlight the potential of azole antifungals as promising platforms that merit further investigation and modification in the pursuit of safer, more effective therapeutics for CDI treatment.

Pancreatic ductal adenocarcinoma (PDAC) remains one of the most aggressive and invasive malignancies, with limited response to the currently available systemic therapy options. Having a 5-year survival of 12%, there is a serious need for novel therapeutic options, including drug repurposing for the treatment of PDAC. In recent years, 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMGCR) inhibitors, also known as statins, have been identified as potential candidates for PDAC therapy repurposing.