This study aims to reveal the effect and mechanism of Gentianella turkestanorum total extract(GTI) in ameliorating non-alcoholic steatohepatitis(NASH). UPLC-Q-TOF-MS was employed to identify the chemical components in GTI. SwissTarget-Prediction, GeneCards, OMIM, and TTD were utilized to screen the targets of GTI components and NASH. The common targets shared by GTI components and NASH were filtered through the STRING database and Cytoscape 3.9.0 to identify core targets, followed by GO and KEGG enrichment analysis. AutoDock was used for molecular docking of key components with core targets. A mouse model of NASH was established with a methionine-choline-deficient high-fat diet. A 4-week drug intervention was conducted, during which mouse weight was monitored, and the liver-to-brain ratio was measured at the end. Hematoxylin-eosin staining, Sirius red staining, and oil red O staining were employed to observe the pathological changes in the liver tissue. The levels of various biomarkers, including aspartate aminotransferase(AST), alanine aminotransferase(ALT), hydroxyproline(HYP), total cholesterol(TC), triglycerides(TG), low-density lipoprotein cholesterol(LDL-C), high-density lipoprotein cholesterol(HDL-C), malondialdehyde(MDA), superoxide dismutase(SOD), and glutathione(GSH), in the serum and liver tissue were determined. RT-qPCR was conducted to measure the mRNA levels of interleukin 1β(IL-1β), interleukin 6(IL-6), tumor necrosis factor α(TNF-α), collagen type I α1 chain(COL1A1), and α-smooth muscle actin(α-SMA). Western blotting was conducted to determine the protein levels of IL-1β, IL-6, TNF-α, and potential drug targets identified through network pharmacology. UPLC-Q-TOF/MS identified 581 chemical components of GTI, and 534 targets of GTI and 1 157 targets of NASH were screened out. The topological analysis of the common targets shared by GTI and NASH identified core targets such as IL-1β, IL-6, protein kinase B(AKT), TNF, and peroxisome proliferator activated receptor gamma(PPARG). GO and KEGG analyses indicated that the ameliorating effect of GTI on NASH was related to inflammatory responses and the phosphoinositide 3-kinase(PI3K)/AKT pathway. The staining results demonstrated that GTI ameliorated hepatocyte vacuolation, swelling, ballooning, and lipid accumulation in NASH mice. Compared with the model group, high doses of GTI reduced the AST, ALT, HYP, TC, and TG levels(P<0.01) while increasing the HDL-C, SOD, and GSH levels(P<0.01). RT-qPCR results showed that GTI down-regulated the mRNA levels of IL-1β, IL-6, TNF-α, COL1A1, and α-SMA(P<0.01). Western blot results indicated that GTI down-regulated the protein levels of IL-1β, IL-6, TNF-α, phosphorylated PI3K(p-PI3K), phosphorylated AKT(p-AKT), phosphorylated inhibitor of nuclear factor kappa B alpha(p-IκBα), and nuclear factor kappa B(NF-κB)(P<0.01). In summary, GTI ameliorates inflammation, dyslipidemia, and oxidative stress associated with NASH by regulating the PI3K/AKT/NF-κB signaling pathway.
Animals ; Non-alcoholic Fatty Liver Disease/genetics* ; Mice ; Network Pharmacology ; Male ; Drugs, Chinese Herbal/administration & dosage* ; Chromatography, High Pressure Liquid ; Liver/metabolism* ; Mice, Inbred C57BL ; Humans ; Mass Spectrometry ; Tumor Necrosis Factor-alpha/metabolism* ; Disease Models, Animal ; Molecular Docking Simulation

AIM: To evaluate the applicability of Chinese dry eye questionnaire in college students using the ocular surface disease index(OSDI)questionnaire as a reference.METHODS: Cross-sectional study. A total of 711 college students from Nanyang Medical College were enrolled in the study and assessed for dry eye condition using OSDI questionnaire and Chinese dry eye questionnaire. The response rate of each question in the two questionnaires was counted. Cronbach α was calculated to evaluate the internal consistency of both questionnaires. Correlation between the total scores of the two questionnaires was analyzed to evaluate the criterion validity. Based on OSDI scores, the discriminant validity of Chinese dry eye questionnaire was evaluated; receiver operating characteristic(ROC)curves was plotted for Chinese dry eye questionnaire scores, area under the ROC curve(AUC)was calculated, and diagnostic thresholds and corresponding sensitivity and specificity were also analyzed.RESULT: The response rates of the 12 questions on the OSDI questionnaire were 33.2%-100.0%, while it was 100.0% for each question on the Chinese dry eye questionnaire. The Cronbach α values of OSDI questionnaire and Chinese dry eye questionnaire were 0.905 and 0.789, respectively. The Chinese dry eye questionnaire score was positively correlated with the OSDI score(rs=0.712, P<0.001). According to OSDI questionnaire scores, dry eye severity was divided into normal group, mild dry eye group, moderate dry eye group and severe dry eye group. The scores of Chinese dry eye questionnaire in these groups were 4.00(2.00, 6.00), 9.00(7.00, 11.00), 12.00(9.00, 14.00)and 16.00(13.50, 22.00), respectively, which increased with the severity of dry eye, and the overall difference was statistically significant(P<0.001), as well as pairwise comparison between groups(P<0.05). The AUCs of Chinese dry eye questionnaire in distinguishing normal population from dry eye population, mild dry eye from moderate dry eye, moderate dry eye from severe dry eye were 0.862, 0.661 and 0.769, respectively, and the diagnostic thresholds were 6.5, 11.5 and 14.5, respectively.CONCLUSION:Chinese dry eye questionnaire has an equivalent reliability, validity, discriminant ability and better response rate for dry eye screening and epidemiological survey among college students in China compared with OSDI questionnaire.

Animals ; Inflammation ; Male ; Myocardial Infarction/metabolism* ; Myocardium/metabolism* ; Oxidative Stress ; Rats ; Rats, Sprague-Dawley

OBJECTIVE: To investigate the effects of Birinapant on hepatocellular carcinoma cells and its related molecular mechanisms. METHODS: Human hepatocellular carcinoma cells QGY-7701 were treated with 0, 1, 5, 25 and 125 nmol/L Birinapant for 24, 48 and 72 hours respectively, each experiment 3 wells.The proliferation activity of cells, the apoptosis levels, the cells nuclear type, the mitochondrial membrane potential, the transcription and expression levels of genes and the cytotoxicity of Birinapant were analyzed.At the same time, 4-week-old male BALB/C mice were randomly divided into 5 groups, with 20 mice in each group.The mice were inguinal injected with QGY-7701 cells, and then subcutaneous injected with Birinapant (concentrations ranging from 0, 1, 5, 25, 125 μg/kg) in each group after two days, once every other day.On 18 day since first Birinapant injection, 10 mice were killed in each group to weigh tumor tissue and survival time was recorded from the remaining 10 mice.The effects of Birinapant on the growth of the tumor and the survival time of tumor-bearing mice were observed. RESULTS: Compared with the negative control (NC) group, the proliferation activity of QGY-7701 was inhibited significantly after Birinapant treatment and the apoptosis levels were increased significantly (<0.01).The cell mitochondrial membrane potential was decreased and the karyotype was changed (<0.01).At the same time, the transcription and expression levels of genes cellular inhibitor of apoptosis protein 1(cIAP-1), cellular inhibitor of apoptosis protein 2(cIAP-2), ras, raf, mek and erk were significantly decreased (<0.01), while the expression levels of caspase-3 and caspase-9 genes were up-regulated (<0.01).Compared with the model group (MG), the growth of the tumor was inhibited significantly and the survival time of the tumor-bearing mice was prolonged after Birinapant treatment (<0.01). CONCLUSIONS Birinapant can inhibit the expression of cIAP-1, cIAP-2 and the proteins of Ras-Raf-MEK-ERK signal pathways, so as to activate the mitochondria mediated endogenous apoptosis pathway.Birinapant shows a certain inhibitory effect on liver cancer.
Animals ; Apoptosis ; Carcinoma, Hepatocellular ; Cell Line, Tumor ; Dipeptides ; Humans ; Indoles ; Liver Neoplasms ; Male ; Mice ; Mice, Inbred BALB C ; Mitochondrial Proteins

OBJECTIVETo evaluate the safety of human umbilical cord derived-mesenchymal stem cell (UC-MSC) transplantation therapy in patients with decompensated liver cirrhosis.

METHODSUC-MSCs were transplanted intravenously into patients with decompensated liver cirrhosis. Serum levels of glucose (GLU), total cholesterol (TC), blood urea nitrogen (BUN), alpha fetoprotein (AFP), white blood cells (WBC), and prothrombin activity (PA) were detected at different time points after UC-MSCs transplantation.

RESULTSMost UC-MSC transplanted patients experienced an improvement in quality of life, to varying degrees. With the exception of low-grade fever in a few patients, side effects and oncogenic events were rare (treatment group: 1/38 vs. control group: 1/16; P more than 0.05). The UC-MSCs transplantation showed no effect on GLU, TC, BUN, AFP, WBC, or PA.

CONCLUSIONUC-MSCs transplantation in patients with decompensated liver cirrhosis is safe and may improve the patient's quality of life.

Adult ; Aged ; Cord Blood Stem Cell Transplantation ; adverse effects ; Female ; Humans ; Liver Cirrhosis ; complications ; surgery ; Male ; Mesenchymal Stem Cell Transplantation ; adverse effects ; Middle Aged ; Prospective Studies