Amygdalin Ameliorates Liver Fibrosis through Inhibiting Activation of TGF-β/Smad Signaling

Objective:To observe the effect of amygdalin on liver fibrosis in a liver fibrosis mouse model,and the underlying mechanisms were partly dissected in vivo and in vitro.Methods:Thirty-two male mice were randomly divided into 4 groups,including control,model,low-and high-dose amygdalin-treated groups,8 mice in each group.Except the control group,mice in the other groups were injected intraperitoneally with 10%carbon tetrachloride(CCl4)-olive oil solution 3 times a week for 6 weeks to induce liver fibrosis.At the first 3 weeks,amygdalin(1.35 and 2.7 mg/kg body weight)were administered by gavage once a day.Mice in the control group received equal quantities of subcutaneous olive oil and intragastric water from the fourth week.At the end of 6 weeks,liver tissue samples were harvested to detect the content of hydroxyproline(Hyp).Hematoxylin and eosin and Sirius red staining were used to observe the inflammation and fibrosis of liver tissue.The expressions of collagenⅠ(Col-Ⅰ),alpha-smooth muscle actin(α-SMA),CD31 and transforming growth factorβ(TGF-β)/Smad signaling pathway were detected by immunohistochemistry,quantitative real-time polymerase chain reaction and Western blot,respectively.The activation models of hepatic stellate cells,JS-1and LX-2 cells induced by TGF-β1 were used in vitro with or without different concentrations of amygdalin(0.1,1,10μmol/L).The effect of different concentrations of amygdalin on the expressions of liver sinusoidal endothelial cells(LSECs)dedifferentiation markers CD31 and CD44 were observed.Results:High-dose of amygdalin significantly reduced the Hyp content and percentage of collagen positive area,and decreased the mRNA and protein expressions of Col-Ι,α-SMA,CD31 and p-Smad2/3 in liver tissues of mice compared to the model group(P<0.01).Amygdalin down-regulated the expressions of Col-Ⅰandα-SMA in JS-1 and LX-2 cells,and TGFβR1,TGFβR2 and p-Smad2/3 in LX-2 cells compared to the model group(P<0.05 or P<0.01).Moreover,1 and 10μmol/L amygdalin inhibited the mRNA and protein expressions of CD31 in LSECs and increased CD44 expression compared to the model group(P<0.05 or P<0.01).Conclusions:Amygdalin can dramatically alleviate liver fibrosis induced by CCl4 in mice and inhibit TGF-β/Smad signaling pathway,consequently suppressing HSCs activation and LSECs dedifferentiation to improve angiogenesis.

Efficacy and Safety of Yanggan Jian in Hepatitis B Virusrelated Decompensated Cirrhosis:A Randomized,Doubleblind,Controlled Trial

Background and Aims:The aim was to evaluate the efficacy and safety of Yanggan Jian(YGJ)in HBV-infected patients with decompensated cirrhosis.Methods:This randomized,double-blind controlled trial enrolled 160 patients with HBV-related decompensated cirrhosis who were already receiving or about to start antiviral therapy.Patients were randomly assigned to receive YGJ or placebo for 24 weeks,and were followed-up to 36 weeks.The primary outcome was the proportion of patients with a≥2 point reduction in Child-Turcotte-Pugh(CTP)score from baseline at week 24.Secondary outcomes were CTP class and score,serum liver function indices,mortality,incidence of hepatocellular carcinoma and variceal bleeding.Results:The proportion of patients with a CTP score reduction≥2 was significantly greater in the YGJ than in the placebo group(p=0.009);the percentage of patients with CTP class C was significantly less than that in the placebo group(p<0.05),and the YGJ group had a significantly greater mean change from baseline in CTP score at week 24(p=0.034).The improvement in measured values and change from baseline of prothrombin time,serum albumin,platelets,cholinesterase,international normalized ratio,and activated partial thromboplastin time were significantly better with YGJ than with placebo.Between-group differences in cumulative rates of variceal bleeding,hepatocellular carcinoma,death,or the frequency of any adverse event(AE),AEs related to treatment,or discontinuation because of AEs were not significant.Conclusions:YGJ significantly improved CTP scores and hepatic synthetic and reserve function in patients with HBV-related decompensated cirrhosis,and was safe and well tolerated.

A novel TICT-based near-infrared fluorescent probe for light-up sensing and imaging of human serum albumin in real samples

Human serum albumin(HSA) has emerged as a pivotal biomarker and prognostic indicator for various human diseases. Real-time sensing and visual tracking of HSA in plasma or other biological systems will immensely facilitate the basic researchers and clinicians to better understand HSA-associated biological processes. Herein, a novel near-infrared(NIR) fluorescent probe(7-HTCF) was rationally constructed for light-up sensing and in-situ imaging of HSA in real samples, based on the principle of twisted intramolecular charge transfer(TICT). Under physiological conditions, 7-HTCF could be efficiently trapped by HSA to form a stable complex via binding on a non-drug binding site, while the complex emitted strong fluoresce signals around 670 nm. Further investigations demonstrated that 7-HTCF displayed a great combination of excellent selectivity and good chemical stability, as well as rapid fluorescent response and ultra-high sensitivity for HSA detection. Particularly, the newly developed light-up probe has been successfully utilized for quantitative detection of HSA in diluted plasma samples, while its readouts are hardly affected by the addition of therapeutic agents and herbal medicines. 7-HTCF is also successfully used for in-situ imaging of the reabsorbed HSA in living renal cells, while this dye exhibits good cell permeability and high resolution for in-situ imaging in living cells. Collectively, a novel TICT-based near-infrared fluorescent probe was devised for highly selective and ultra-sensitive sensing of HSA in plasma samples or imaging HSA in living cells, which offered a practical tool for clinical tests and for exploring HSA-associated biological processes.

Comprehensive profiling and characterization of the absorbed components and metabolites in mice serum and tissues following oral administration of Qing-Fei-Pai-Du decoction by UHPLC-Q-Exactive-Orbitrap HRMS

Qing-Fei-Pai-Du decoction(QFPDD)is a Chinese medicine compound formula recommended for combating corona virus disease 2019(COVID-19)by National Health Commission of the People's Republic of China.The latest clinical study showed that early treatment with QFPDD was associated with favorable outcomes for patient recovery,viral shedding,hospital stay,and course of the disease.However,the effective constituents of QFPDD remain unclear.In this study,an UHPLC-Q-Orbitrap HRMS based method was developed to identify the chemical constituents in QFPDD and the absorbed prototypes as well as the metabolites in mice serum and tissues following oral administration of QFPDD.A total of 405 chemicals,including 40 kinds of alkaloids,162 kinds of flavonoids,44 kinds of organic acids,71 kinds of triterpene saponins and 88 kinds of other compounds in the water extract of QFPDD were tentatively identified via comparison with the retention times and MS/MS spectra of the standards or refereed by literature.With the help of the standards and in vitro metabolites,195 chemical components(including 104 prototypes and 91 metabolites)were identified in mice serum after oral administration of QFPDD.In addition,165,177,112,120,44,53 constituents were identified in the lung,liver,heart,kidney,brain,and spleen of QFPDD-treated mice,respectively.These findings provided key information and guidance for further investigation on the pharmacologically active substances and clinical applications of QFPDD.

Altered oral microbiota in chronic hepatitis B patients with different tongue coatings

AIM To elucidate tongue coating microbiota and metabolic differences in chronic hepatitis B(CHB) patients with yellow or white tongue coatings.METHODS Tongue coating samples were collected from 53 CHBpatients(28 CHB yellow tongue coating patients and 25 CHB white tongue coating patients) and 22 healthy controls.Microbial DNA was extracted from the tongue samples,and the bacterial 16 S ribosomal RNA gene V3 region was amplified from all samples and sequenced with the Ion Torrent PGM^(TM)sequencing platform according to the standard protocols.The metabolites in the tongue coatings were evaluated using a liquid chromatographymass spectrometry(LC-MS) platform.Statistical analyses were then performed.RESULTS The relative compositions of the tongue coating microbiotas and metabolites in the CHB patients were significantly different from those of the healthy controls,but the tongue coating microbiota abundances and diversity levels were not significantly different.Compared with the CHB white tongue coating patients,the CHB yellow tongue coating patients had higher hepatitis B viral DNA(HBV-DNA) titers(median 21210 vs 500,respectively,P = 0.03) and a significantly lower level of Bacteroidetes(20.14% vs 27.93%,respectively,P = 0.013) and higher level of Proteobacteria(25.99% vs 18.17%,respectively,P = 0.045) in the microbial compositions at the phylum level.The inferred metagenomic pathways enriched in the CHB yellow tongue coating patients were mainly those involved in amino acid metabolism,which was consistent with the metabolic disorder.The abundances of bacteria from Bacteroidales at the order level were higher in the CHB white tongue coating patients(19.2% vs 27.22%,respectively,P = 0.011),whereas Neisseriales were enriched in the yellow tongue coating patients(21.85% vs 13.83%,respectively,P = 0.029).At the family level,the abundance of Neisseriaceae in the yellow tongue patients was positively correlated with the HBV-DNA level but negatively correlated with the S-adenosyl-L-methionine level.CONCLUSION This research illustrates specific clinical features and bacterial structures in CHB patients with different tongue coatings,which facilitates understanding of the traditional tongue diagnosis.

Research Progress in Chinese Medicine Preparations for Promoting Blood Circulation and Removing Blood Stasis for Cirrhotic Patients with Portal Vein Thrombosis Following Splenectomy

This article presented an overview of the therapeutic effects of Chinese medicine(CM)preparations for promoting blood circulation and removing blood stasis for patients with portal vein thrombosis(PVT)after splenectomy.Based on published clinical researches of CM preparations for PVT after splenectomy in patients with cirrhotic portal hypertension(CPH),this paper evaluated the incidence of PVT,and explored potential active components and mechanisms of CM preparations.Safflower Yellow Injection,Danshen Injection,Danhong Injection,and Compound Danshen Dropping Pill achieved good curative effect alone or combined with anticoagulant therapy.In addition,Compound Biejia Ruangan Tablet and Anluo Huaxian Pill can also significantly improve the hemodynamic disorders of portal vein system in patients with cirrhosis.Considering the role of CM preparations in ameliorating the incidence of PVT after splenectomy in patients with CPH,we suggested that future research should provide more attention to CM alone or CM combined with anticoagulant for cirrhosis with PVT.