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Abstract Background Oxymatrine (OM), a quinolizidine alkaloid extracted from a herb Sophorae Flavescentis Radix, has been used to treat liver fibrotic diseases. However, the mechanism of its anti-fibrosis effects is still unclear. TGF-β/Smad signaling and miR-195 have been proved to paly an important role in hepatic stellate cells (HSCs) activation and liver fibrosis. In this study, we investigated whether OM could inhibit HSCs activation through TGF-β1/miR-195/Smads signaling or not. Methods First, the effects of OM on HSC-T6 in different concentrations and time points were tested by MTT assay. We choose three appropriate concentrations of OM as treatment concentrations in following experiment. By Quantitative Real-time PCR and Western Blot, then we investigated the effect of OM on miR-195, Smad7 and α-SMA’s expressions to prove the correlation between OM and the TGF-β1/miR-195/Smads signaling. Last, miR-195 mimic and INF-γ were used to investigate the relation between miR-195 and OM in HSC activation. Results Our results showed that the proliferation of HSC was significantly inhibited when OM concentration was higher than 200 μg/mL after 24 h, 100 μg/mL after 48 h and 10 μg/mL after 72 h. The IC50 of OM after 24, 48 and 72 h were 539, 454, 387 μg/mL respectively. OM could down-regulate miR-195 and α-SMA (P

OBJECTIVES: We aim to obtain the intra-subject coefficient of variability of a highly variable antidepressant agomelatine in humans, and propose an adjusted bioequivalence assessment strategy. METHODS: A single-dose, randomized crossover design was conducted in four periods (reference administered thrice, placebo administered once) separated by seven days. A validated LC-MS/MS assay was used to measure drug concentrations in serial blood samples. RESULTS: The intra-subject coefficient of variability was calculated using the residual variance of ANOVA analysis, and the results for Cmax and AUC0-t was 78.34% and 43.52%, respectively, in Chinese healthy subjects. The sample size required for standard BE study were 124(192, 340) if the expected deviation between the reference and generic products was set to 0 (5%, 10%). CONCLUSIONS: Agomelatine meets the criteria for highly variable drug in Chinese healthy male subjects, and the traditional BE criteria for agomelatine needs to be adjusted to alleviate the resource and ethical burden of using a large numbers of subjects in clinical trials. Our clinical data on the intra-subject variability of agomelatine PK in Chinese healthy population enables to adjust bioequivalence (BE) assessment approach for agomelatine based on the RSABE approaches recommended by regulatory agencies. TRIAL REGISTRATION: ChiCTR.org ChiCTR-TTRCC-13003835.

Background and Aim. Aberrant activationof theTGF-𝛽1/Smad pathway contributes to the activation of hepatic stellate cells (HSCs). MicroRNA-195 has been shown to regulate the activation of HSCs.Theaim of this study was to investigate the role of miRNA-195 in HSCs activation. Methods. A liver fibrotic ratmodel induced by diethylnitrosamine was established. Dual luciferase reporter assays were performed to verify that Smad7 was the target of miRNA-195.The expression levels of miR-195, Smad7, and 𝛼-SMAin HSC-T6 transfected, respectively, with miR-195 mimic, inhibitor, or control were measured by qRT-PCR. The protein expression of Smad7 was detected by Western blot analysis. Results. Enhanced miR-195 and decreased Smad7 were observed in diethylnitrosamineinduced liver fibrotic rats (𝑃 < 0.05). Dual luciferase reporter assays showed that the miR-195 mimic significantly suppressed the luciferase activity of a reporter plasmid carrying the binding site of miR-195 on the 3󸀠UTR of Smad7 (𝑃 < 0.05).ThemiR-195 mimics activated HSCs, further elevated miR-195 and 𝛼-SMA (𝑃 < 0.01), and reduced the Smad7 level (𝑃 < 0.05). The miR-195 inhibitors blocked the activation of HSCs, reduced the expression of miR-195 and 𝛼-SMA (𝑃 < 0.01), and upregulated the expression of Smad7 (𝑃 < 0.05). Conclusion. Collectively, we demonstrated that miRNA-195 activated HSCs by targeting Smad7. [ABSTRACT FROM AUTHOR]

Probe 1 was designed and synthesized as a new fluorescent molecular probe for thiols in PBS buffer at physiological condition. This fluorescent molecular probe consists of a thiol reaction moiety bound to a coumarin fluorophore. Its fluorescence quantum yield is low, but a drastic enhancement of fluorescence intensity was observed in the presence of thiols. Possible interference with other analytes was examined. Probe 1 displays a highly selective fluorescent enhancement with thiols, and the probe was successfully applied to thiols determination in intracellular, in human urine and blood samples. [ABSTRACT FROM AUTHOR]