Rapamycin is frequently used as an immunosuppressive agent and also plays an important role in tumor suppression.It combines to FKBP12 and forms a compound to inhibit the serine/threonine protein kinase mammalian target of rapamycin(mTOR),its activation is required for protein synthesis and cell-cycle progression.Rapamycin can block cell cycle at G1 phase and to inhibit cell proliferation.Furthermore rapamycin may inhibit cytokine production and cytokine signaling.Therefore rapamycin is used in the treatment of post-transplantation organ rejection and tumor therapy due to its dual effects of immuno-suppression and tumor inhibition.

ObjectiveTo explore the association between hepatic steatosis and the liver tissue expression of HBsAg and HBcAg in chronic hepatitis B (CHB) patients.MethodsFrom January 2005 to June 2008,a total of 147 CHB patients with hepatic steatosis diagnosed by liver biopsy and other 149 CHB patients without hepatic steatosis but with similar HBV DNA titer were enrolled.The differences of HBsAg and HBcAg immunostaining and liver injury in these two groups were compared.The data were analysed using t test and chi square test.ResultsCompared with non-steatosis group,the average age and body weight index of hepatic steatosis group were higher (t values were -3.31and -6.57,both P＜0.01).The percentage of moderate to severe hepatic inflammation in liver,obvious hepatic fibrosis and the strong positive HBsAg staining was lower (30.6％ vs 15.4％ ； 26.5％vs 12.8％； 23.1 ％ vs 6.7 ％； x2=9.63,8.92,15.76； all P＜0.01),and the percentage of strong positive HBcAg staining was also in downtrend.Compared with degree F1 and F2 of liver steatosis,the percentage of HBsAg and HBcAg strong positive staining in liver tissues of degree F3 and F4 was in downtrend.ConclusionsHepatic steatosis affected the expression of HBsAg and HBcAg in liver tissue of CHB patients.As hepatic steatosis appeared and became more severe,both expression of HBsAg and HBcAg and the degree of liver injury were in downtrend.

Diagnosis, Differential ; Humans ; Liver Diseases, Alcoholic ; diagnosis ; Non-alcoholic Fatty Liver Disease ; diagnosis

Objective:To establish a real-time fluorescent quantitative PCR for the detection of torque teno virus types 7 (TTV7), 8 (TTV8) and 10 (TTV10) and analyze its performance in clinical sample detection.Methods:Specific primers were designed based on the gene sequences of TTV7, TTV8 and TTV10 in GenBank. Recombinant plasmids of pMD19-T-TTV7, pMD19-T-TTV8 and pMD19-T-TTV10 were constructed and used as positive standard control to establish a real-time fluorescent quantitative PCR based on FAM-Eclipse probe method. The specificity and sensitivity of the established method were evaluated. Moreover, it was validated in terms of clinical sample detection.Results:The standard curve equations of the real-time fluorescent quantitative PCR for detecting TTV7, TTV8 and TTV10 were y=-0.340 2 x+ 114.780 0 ( R2=0.998 8), y=-0.351 1 x+ 114.940 0 ( R2=0.995 3) and y=-0.348 9 x+ 115.020 0 ( R2=0.991 7), respectively, and there was no cross-reaction with other viruses. The detection sensitivity of the established method for TTV7, TTV8 and TTV10 were 108 copies/μl, 84 copies/μl and 98 copies/μl, and the positive detection rates in clinical pediatric serum samples were 10.9%, 2.1% and 4.3%, respectively. Conclusions:The established real-time fluorescent quantitative PCR for detection of TTV7, TTV8 and TTV10 was featured by strong specificity and high sensitivity, which could be used for rapid TTV detection in clinical serum samples.

Objective To investigate the differences and similarities between drug-induced liver injury (DILI) and autoimmune hepatitis (AIH) in serum biochemical parameters and liver pathology,and to provide some thoughts for clinical diagnosis and differentiation of these two diseases.Methods A retrospective analysis was performed for the biochemical,immunological,autoantibody,and liver pathological data of 106 DILI patients and 63 AIH patients who were hospitalized,diagnosed,and treated in our hospital from January 2012 to October 2014.The patients' general data,biochemical parameters,immunological data,Ishak score,and qualitative changes in liver tissue were analyzed and compared.The Kruskal-Wallis test was used for comparison of nonparametric data between multiple groups,the Nemenyi test was used for comparison of nonparametric data between any two groups,the Wilcoxon rank sum test was used for comparison of Ishak scores,and the chi-square test was used for comparison of constituent ratio of categorical data.Results There were significant differences between AIH group and DILI hepatocyte injury group/mixed-type DILI group in the following serum biochemical parameters:alanine aminotransferase (187.2 U/L vs 1 326.5 U/L and 455.6,P ＜ 0.05),aspartate aminotransferase (172.2 U/L vs 759.5 U/L and 349.5 U/L,P ＜0.05),alkaline phosphatase (209.3 U/L vs 157.3 U/L and 169.4 U/L,P ＜ 0.05),gamma-glutamyl transferase (254.8 U/L vs 176.5 U/L and 170.5 U/L,P ＜ 0.05),total bilirubin (37.2 μmol/L vs 95.8 μmol/L and 52.6 μmol/L,P ＜ 0.05),serum iron (18.9 μmol/L vs 36.2 μmol/L and 23.9 μmol/L,P ＜ 0.05),serum ferritin (122.5 μmol/L vs 410.4 μmol/L and 186.5 μmol/L,P ＜ 0.05),immunoglobulin G (18.4 g/L vs 12.6 g/L and 12.3 g/L,P ＜ 0.05),and immunoglobulin M (1.8 g/L vs 1.3 g/L and 1.1 g/L,P ＜ 0.05).There were also significant differences between AIH group and DILI hepatocyte injury group/mixed-type DILI group in the Ishak score for interface inflammation (2.2±0.8 vs 1.3±0.7 and 1.3±0.6,P ＜ 0.05),Ishak score for portal inflammation (2.3±0.9 vs 1.5±0.7 and 1.4±0.8,P ＜ 0.05),and fibrosis score (2.8±1.1 vs 1.5±0.7 and 1.3±0.7,P ＜ 0.05).There were significant differences between AIH group and DILI hepatocyte injury group/mixed-type DILI group in the proportion of wax-like deposition (0 vs 29.2％ and 34.5％,P ＜0.05) and proportion of iron deposition (11.1％vs 52.1％ and 25.9％,P ＜ 0.05).Conclusion There are differences in biochemistry,immunology,and liver histology between DILI and AIH patients.AIH patients have more serious interface inflammation and portal inflammation and a higher fibrosis degree compared with DILI patients,while DILI patients have greater proportions of wax-like deposition and iron deposition compared with AIH patients.

BACKGROUND/AIMS: This study aimed to verify the reliability of the alcoholic liver disease (ALD)/nonalcoholic fatty liver disease (NAFLD) index (ANI) for distinguishing ALD in patients with hepatic steatosis from NAFLD, and to investigate whether ANI combined with γ-glutamyl transferase (GGT) would enhance the accuracy of diagnosis in China. METHODS: A hundred thirty-nine cases of fatty liver disease (FLD) were divided into two groups of ALD and NAFLD. The ANI was calculated with an online calculator. All indicators and ANI values were analyzed using statistical methods. RESULTS: ANI was significantly higher in patients with ALD than in those with NAFLD (7.11 ± 5.77 vs. -3.09 ± 3.89, p < 0.001). With a cut-off value of -0.22, the sensitivity, specificity, and area under the receiver operating characteristic curve (AUROC) of diagnosed ALD cases was 87.1%, 92.5%, and 0.934 (95% confidence interval [CI], 0.879 to 0.969), respectively. The corresponding values for aspartate aminotransferase (AST)/alanine transaminase (ALT), mean corpuscular volume (MCV), and GGT were 75.29%, 72.94%, and 0.826 (95% CI, 0.752 to 0.885); 94.34%, 83.02%, and 0.814 (95% CI, 0.739 to 0.875) and 80.23%, 79.25%, and 0.815 (95% CI, 0.740 to 0.876), respectively. ANI AUROC was significantly higher than the AST/ALT, MCV, or GGT AUROCs (all p < 0.001), moreover, ANI showed better diagnostic performance. The combination of ANI and GGT showed a better AUROC than ANI alone (0.976 vs. 0.934, p = 0.016). The difference in AUROCs between AST/ALT, MCV, and GGT was not statistically significant (all p > 0.05). CONCLUSIONS: ANI can help distinguish ALD from NAFLD with high accuracy; when ANI was combined with GGT, its effectiveness improved further.
Alcoholics* ; Aspartate Aminotransferases ; China ; Diagnosis ; Diagnosis, Differential ; Erythrocyte Indices ; Fatty Liver* ; gamma-Glutamyltransferase ; Humans ; Liver Diseases, Alcoholic* ; ROC Curve ; Sensitivity and Specificity ; Transferases*

Objective:To investigate the influencing factors of significant liver fibrosis in patients with chronic hepatitis B (CHB) concurrent with non-alcoholic fatty liver disease (NAFLD).Methods:Those who underwent liver pathological examination and confirmed diagnosis of CHB and NAFLD in Tianjin Second People′s Hospital from August 2014 to September 2017 were enrolled. Data regarding their demographic information, laboratory tests results, and liver pathology results were analyzed. The latter results were used to categorize the patients either in non-significant liver fibrosis group (Metavir stage

Objective To investigate the expression level of fatty acid synthase (FASN) in different grades ofgliomas and its relationship with glioma malignancy.Methods Glioma U87 and U373 cell lines were routinely cultured in vitro;immunofluorescence assay was performed to detect the cellular localization of FASN in glioma U87 and U373 cell lines.Sixty-seven human glioma samples,collected in our hospital from March 2012 to March 2013,were used in our study;grade Ⅰ was noted in three samples by WHO grading,grade Ⅱ in 29,graded Ⅲ in 22,and grade Ⅳ in 13;immunohistochemistry was performed to detect the expression levels of FASN,CD34 and Ki-67 and microvascular density (MVD) in these human glioma samples.Spearman's correlation was used to analyze the relationship between FASN expression and both Ki-67 expression and MVD in human gliomas.Results (1) FASN could express in the cytoplasm of glioma U87 and U373 cell lines.(2) immumohistochemical staining indicated that FASN mainly located in the cytoplasm,Ki-67 in the cell nucleus,and CD34 in the cytomembrane.FASN expression in samples of grade Ⅰ (3.3683±0.6549) was significantly lower than that in samples of grade Ⅱ (4.0512±0.4859,P＜0.05),and FASN expression in samples of grade Ⅲ (4.1881±0.5755) was significantly lower than that in samples of grade Ⅳ (4.6996±0.5164,P＜0.05);MVD in samples of grade Ⅱ was significantly smaller than that in samples of grade Ⅲ(P＜0.05),and MVD in samples of grade Ⅲ was significantly lower than that in samples of grade Ⅳ(P＜0.05);Ki-67 expression in samples of grade Ⅱ was significantly lower than that in samples of grade Ⅲ (P＜0.05).(3) There was a positive and significant correlation between FASN expression and MVD (r=0.606,P=0.030);a positive and significant correlation between FASN expression and Ki-67 was also noted (r=0.636,P=0.014).Conclusion High expression level of FASN indicates high proliferation capability and high blood supply in haman gliomas.

Objective:A mouse model of pancreatitis induced by coxsackievirus B3 (CVB3) was established. The pathological change of pancreas and the infiltration of Th17/Treg cells were observed.Methods:The BALB/c mice were inoculated intraperitoneally with CVB3 to induce acute viral pancreatitis model. Then the pathological changes of pancreas were observed by HE staining; the viral RNA load and relative expression of cytokines (IFN-γ, IL-6 and IL-17) mRNA were detected by q-PCR; the proportion of infiltrated CD45 + CD3 + T cells, CD4 + and CD8 + T cells, Th17 and Treg cells in the pancreas was determined by flow cytometry. Results:Three days after CVB3 infection, the viral RNA load in pancreas was the highest (0.96±0.18) and gradually decreased with prolongation of infection. Compared with the 3 dpi group, the viral RNA load in pancreas was decreased (0.96±0.18 vs. 0.62±0.14) at 7 dpi, but there was no statistically significant difference. In addition, the infiltration of immune cell in pancreas increased significantly after 7dpi and the pathological score >2. The percent of infiltrated Th17 cells (1.05±0.21 vs. 22.13±5.79) and Treg cells (3.11±0.78 vs. 8.25±1.30) among CD4 + T cells significantly increased after infection (P<0.05), and the Th17/Treg also increased (P<0.01). Compared with the control group, the relative mRNA expression of IFN-γ (1.05±0.23 vs. 672.6±47.67), IL-6 (1.00±0.38 vs. 68.28±4.57), and IL-17 (1.01±0.11 vs. 54.15±7.94) in pancreas increased at 7 days after CVB3 infection ( P<0.01). Conclusions:The infiltration of Th17/Treg cells and the expression of related cytokines related cytokines IL-6 and IL-17 mRNA were upregulated in pancreas, which promoted the process of CVB3-induced pancreatitis.