Oral anti-viral agents nucleos (t)ide analogues (NAs) are widely used in treatment of chronic hepatitis B (CHB) since they are well-tolerated in most patients.But clinical relapse occurs in a considerable part of patients after the cessation of NAs therapy.This paper reviews factors related to clinical relapse, including time for drug withdrawal, cccDNA level, age, baseline alanine aminotransferase (ALT) and HBV DNA levels, type of NAs, serum level of HBsAg and the immune status of patients.

MicroRNAs (miRNAs) are small non-coding RNAs that regulate both mRNA and protein expression of target genes and play important roles in proliferation,differentiation,development and metabolism of cells.This paper reviews the research progress on miRNAs involvement in liver diseases,including viral hepatitis,fatty liver,drug induced liver disease,primary biliary cirrhosis and primary hepatocellular carcinoma.

OBJECTIVE To investigate the infection and drug resistance of Staphylococcus spp from hospitalized cases in Suzhou area.METHODS The data from hospitalized cases of 32 hospitals in Suzhou(from 2004 to 2007) were analyzed retrospectively.RESULTS From 2004 to 2007,17 668 cases of nosocomial infection were collected from 32 hospitals in Suzhou area.The infection rate of Staphylococcus aureus was 5.78%,7.11%,8.39% and 7.50%,respectively;the number of meticillin-resistant S.aureus(MRSA) infection cases was 66(34.74%),107(33.86%),138(37.00%) and 219(53.16%) respectively and the total number was 530(41.05%).The nosocomial infection caused by S.epidermidis accounted for 5.99%,5.47%,5.35% and 5.25%,respectively from 2004 to 2007.The number of meticillin-resistant S.epidermidis(MRSE) infection cases were 118(59.90%),128(52.67%),119(50.00%) and 134(46.53%) and the total number was 499(51.66%).The main infection site of S.aureus and S.epidermidis was respiratory tract(74.28% and 71.81%,respectively).Antibiotic resistance strains of S.aureus and S.epidermidis was on rising,and most of them were multi-drug resistance strains.All of the strains were sensitive to vancomycin.CONCLUSIONS In Suzhou area,nosocomial infection and drug resistance of Staphylococcus is on the rise.Evevy hospital must take effective measures to control nosocomial infections of Staphylococcus and drug resistance.

This study sought to isolate and purify C-kit+ cells from rat 2-AAF/PH model and to investigate the proliferation and differentiation of C-kit+ cells in vitro. C-kit positive oval cells were enriched by using magnetic activated cell sorting (MACS). The sorted oval cells were cultured in a low density, and then colony formation was observed. The capacity of proliferation and differentiation of C-kit positive cells were examined in vitro by immunocytochemistry and RT-PCR. By using C-kit antibody in conjunction with MACS, we developed a rapid oval cell isolation protocol. The sorted cells formed colony when cultured in vitro. Cells in the colony expressed albumin or cytokeratin 19 (CK19) or coexpressed both, and BrdU incorporation test was positive. RT-PCR on colony showed expression of albumin and CK19 gene. The results demonstrate that by means of MACS we have established a method to isolate oval cells. The sorted hepatic oval cells can form colony in vitro which expresses different combinations of phenotypic markers and genes from both hepatocytes and cholangiocyte lineage.
2-Acetylaminofluorene ; pharmacology ; Animals ; Animals, Newborn ; Cell Differentiation ; Cell Proliferation ; Cells, Cultured ; Hepatocytes ; cytology ; metabolism ; Male ; Proto-Oncogene Proteins c-kit ; biosynthesis ; genetics ; Rats ; Rats, Sprague-Dawley ; Stem Cells ; cytology ; metabolism

Objective To observe the expression level of macrophage stimulating protein (MSP) in acute on-chronic liver.failure (ACLF) patients,and to explore the clinical significance and correlation with different immune regulatory factors.Methods The double antigen sandwich enzyme-linked immunosorbent assay method was used to detect MSP in the peripheral blood of 45 patients who were diagnosed with ACLF and 32 cases of chronic hepatitis B (CHB).The MSP levels were compared among ACLF patients with different outcomes,and the MSP level of healthy person was used as control.Meanwhile,liver function and hepatitis B virus (HBV) load were detected,and the expressions of peripheral blood CD4+ interferon (IFN)γ+ (helper T cell 1 Th1),CD4+ interleukin (IL)-4+ (helper T cell 2,Th2),CD4+IL-17+ (helper T cell 17,Th17) and CD4+ CD25+ Foxp3+ (regular T cell,Treg) were measured by flow cytometry.The comparison of means between two samples was done by t test,and oneway ANOVA and linear correlation analysis were also used.Results The serum MSP levels in ACLF patients,CHB group and healthy control were (1.65±0.46) ng/mL,(1.43±0.32) ng/mL and (1.23±0.21) ng/mL,respectively.The serum MSP level in ACLF patients was significantly higher than both CHB patients (t=2.163,P=0.035) and healthy control (t=4.032,P=0.01).The MSP level in ACLF survival group was statistically higher compared with ACLF death group ([2.29 ± 0.42] ng/mL vs [1.42±0.17] ng/mL,t=1.973,P=0.042).Th2,Th17 cells in ACLF group,CHB group and healthy control group were (1.51±0.27) ％ and (1.94±1.02)％,(0.42±0.08)％ and (0.55±0.36)％,(0.23±0.19) ％ and (0.26±0.19) ％,respectively,which were all significantly different (F=7.759 and 37.229,respectively;both P＜0.01).The MSP level was positively associated with the number of Th2 (r=0.386,P=0.032) and Th17 (r=0.644,P=0.000),and the ratio of Th17/Treg (r =0.605,P=0.000);while it was negatively associated with the number of Th1 (r=-0.212),Treg (r=-0.262) and the ratio of Th1/Th2 (r=-0.394) (all P＞0.05).Conclusion MSP is involved in the progress of ACLF,and it may be associated with clinical outcomes and cellular immune imbalance of ACLF patients.

Objective To explore the significance and the change of miRNAs expression profile in different period of HBV infection.Methods Establish the detection method of microRNA(miRNA) by RNA DNA probe liquid chip (flexible multi-analyte profiling,xMap) and estimate the specificity,repeatability and accuracy of this method.From October 2010 to October 2011,collect HBV infected patients' periphcral blood of the First affiliated hospital of SooChow University,including acute hepatitis B,chronic hepatitis B,hepatits B liver cirrhosis,liver cancer patients and health control,each group contains 40 cases.The levels of miR-191,-223,-222,-145,-21,-31,-126,-20a,-372 of peripheral blood mononuclear cell were detected by xMap liquid chiptechnology.The level of miR-103 was taken as reference.The ratio of (the mean fluorescence intensity of target miRNAs-corresponding backgroud mean fluorescence intensity)/( the mean fluorescence intensity of miR-103-corresponding backgroud mean fluorescence intensity) as a valid data and analysis the characteristics of miRNAs expression.The SPSS 17.0 was used as statistical software.The single factor analysis of variance was used as the method to analysis group comparison,and make multiple comparison by the LSD-t method if the result with a significant difference.Results The specificity of Xmap liquid chip method to detect miRNAs was 100％ ;The repeated experiment proved that the CV value was less than 5％ in the high value reference miRNA test,less than 10％ in the low value reference miRNAs test;the accruracy experiment proved that the recovery rate was ( 100 ± 5)％ in the nine miRNAs.There were no statistically differences with miR-222 ( F =1.32,P ＞ 0.05),-191 ( F =1.98,P ＞ 0.05),-145 ( F =0.78,P＞0.05),-21(F=0.64,P ＞0.05),-31 (F =0.83,P ＞0.05),-372(F =1.75,P ＞0.05)in different groups; There was statistically significant differences in miR-223 ( F =14.56,P ＜ 0.05) among different groups,with the highest expression level in actue hepatitis B group (15.37 ± 4.01),and the lowest expression level in liver cancer group (6.91 ±3.18) ; There was statistically significant differences in miR126 (F =17.43,P ＜ 0.05)among different groups,with the highest expression level in health control group (6.33 ±2.75) and the lowest expression level in liver cancer group (2.38 ± 1.07).There were statistically significant differences in miR-20a ( F =19.484,P ＜ 0.05) among different groups,with the highest expression level in health control group (0.33 ±0.18) and the lowest expression level in liver cancer group (0.81 ±0.24).Conclusion The detection method of miRNA by Xmap liquid chip has strong specificity,high accuracy and good repeatability,suitable for large throughput clinical testing.The study for miRNA in HBV infected diseases provides a new clue to the research of chronic progress mechanism.

Objective To establish and assess a new screening method for anti-HIV-1 drugs.Methods JLTRG cells were co-cultured with different proportions of H9/HTLV-Ⅲ B cells for 24,48,72 and 96 h.Intensity and density of green fluorescent protein were observed under fluorescence microscope,and were tested using flow cytometry.The optimal proportion of cells in co-culture system and the culture time were determined.The effectiveness of Enfuvirtide (T20) and Efavirenz (EFV),and their half maximal inhibitory concentrations (IC50) were determined by using cell co-culture system and half life of drugs.HIV load was detected using RT-PCR for HIV-1 p24 antigen,and its correlations with drug concentration and mean fluorescent intensity were analyzed by Spearman rank correlation analysis.Results Experiments demonstrated that JLTRG cells co-cultured with H9/HTLV-ⅢB cells at the proportion of 10 ∶ 1 for 72 hours was the best.Along with the concentrations of T20 and EFV changed,JLTRG cells were infected with HIV-1 in different degrees,and the IC50s of T20 and EFV were 10 nmol/L and 5 nmol/L,respectively.The concentrations of T20 and EFV were negatively correlated with mean fluorescent intensity and viral load (r =-1,-0.986 and-1,-1,P ＜ 0.01); and mean fluorescent intensity was positively correlated with viral load (r =0.986 and 1,P ＜ 0.01).Conclusion The drug screening method established in this study is efficient and easy to operate,which provides a new option for anti-HIV-1 drug screening.

Objective To investigate the changes and clinical significance of peripheral blood T lymphocyte subsets and cytokines in influenza A (H7N9) virus infection patients.Methods Twenty-three patients with H7N9 virus infection who received treatment at the First Affiliated Hospital of Soochow University from April 2013 to April 2015 were enrolled as case group.Twenty healthy subjects in the outpatient clinic during the same period were selected as control group.The percentages of T lymphocyte subsets in the peripheral blood including CD3+CD4+ T cells, CD3+CD8+ T cells, regulatory T lymphocytes (Treg), Th17, CD3+CD8-interferon (IFN)-γ+ (Th1) cells and CD3+CD8-IL-4+ (Th2) cells were detected by flow cytometry.The changes of plasma cytokines including interleukin (IL)-8, IL-18, interferon-inducible protein (IP)-10, macrophage inflammatory protein (MIP)-1b were quantified by Bio-Plex Pro human cytokine Group Ⅰ 27-plex panel and Group Ⅱ 21-plex panel.The normality test was performed by Kolmogorov-Smirnov test.Two independent samples t test were used to compare the two groups.Results The percentages of CD3+CD4+ and CD3+CD8+ T cells before treatment in case group were significantly lower than control group ([27.90±10.19]% vs [38.75±6.78]%, t=-2.726, P=0.012;[14.82±7.72]% vs [22.79±6.12]%, t=-2.556, P=0.018), while the percentages of Th17 and Th2 before treatment in case group were significantly higher than control group ([2.64±1.40]% vs [0.29±0.21]%, t=4.668, P<0.001;[2.24±2.00]% vs [0.35±0.25]%, t=2.626, P=0.014).After treatment, 15 cases achieved improvement, among which the percentages of CD3+CD4+ T cells, CD3+CD8+ T cells and Treg after treatment were significantly higher than those before treatment ([36.54±9.20]% vs [25.86±7.22]%, t=-3.339, P=0.007;[22.70±5.68]% vs [15.08±8.80]%, t=-2.811, P=0.017;[6.08±1.70]% vs [3.26±1.64]%, t=-6.670, P<0.001), while the percentages of Th17 and Th1 cells after treatment were significantly lower than those before treatment ([2.12±1.28]% vs [2.76±1.33]%, t=2.220, P=0.048;[4.00±2.13]% vs [6.97±1.81]%, t=4.407, P=0.001).A total of 8 patients died with no significance differences of all the above mentioned immunological parameters after treatment compared with before treatment (all P>0.05).Before treatment, levels of IL-8, IL-18, IP-10 and MIP-1b of case group were significantly higher than those in control group (IL-8: [23.19±14.35] vs [12.78±6.76] ng/L, t=2.277, P=0.035;IL-18:[230.55±230.18] vs [72.80±27.91] ng/L, t=2.348, P=0.036;IP-10:[28 870.55±41 815.22] vs [1 356.13±1 093.10] ng/L, t=2.371, P=0.035;MIP-1b: [197.74±119.87] vs [118.51±41.86] ng/L, t=2.198, P=0.043).Conclusions Patients with H7N9 virus infection exhibit an imbalance of T lymphocyte subsets.It is very important to monitor the changes of T lymphocyte subsets in those patients for clinical prognosis.A storm of cytokines could exist during H7N9 virus infection, which may be the main reason for multiple organ failure.

Hepatitis B ; Humans ; Th17 Cells

ObjectiveTo investigate the value of Model for End-Stage Liver Disease (MELD) score combined with platelet-to-white blood cell ratio (PWR) in predicting the short-term prognosis of patients with hepatitis B virus-associated acute-on-chronic liver failure (HBV-ACLF). MethodsA retrospective analysis was performed for the clinical data of 123 HBV-ACLF patients who were admitted to The First Affiliated Hospital of Suzhou University from June 2014 to June 2019, and according to the prognosis on day 90 after admission, these patients were divided into survival group with 53 patients and death group with 70 patients. Related clinical data were recorded, including age, sex, and total bilirubin (TBil), alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma-glutamyl transpeptidase (GGT), alkaline phosphatase (ALP), serum creatinine (SCr), Albumin (Alb), prealbumin (PAB), international normalized ratio (INR), white blood cell count (WBC), lymphocyte count (LY), monocyte count (MO), neutrophil count (NE), hemoglobin (Hb), and platelet count (PLT) within 24 hours after admission, and PWR and MELD score were calculated. The t-test and the Mann-Whitney U test were used for comparison of continuous data between two groups; univariate and multivariate binary logistic regression analyses were used to analyze the association between each factor and the prognosis of HBV-ACLF; a predictive model of MELD score combined with PWR was established. The receiver operating characteristic (ROC) curve was plotted, and Youden index, cut-off value, sensitivity, and specificity were calculated; the area under the ROC curve (AUC) was calculated for MELD score alone or combined with PWR to compare their value in predicting the prognosis of HBV-ACLF patients. ResultsThere were significant differences between the two groups in TBil, ALT, SCr, INR, WBC, MO, NE, Hb, PLT, PWR, and MELD score (all P＜0.05). TBil, SCr, INR, WBC, MO, NE, and MELD score were risk factors for prognosis of HBV-ACLF patients(all P＜0.05); PWR (odds ratio ［OR］=0.883, 95% confidence interval ［CI］: 0.798-0.977, P=0.016) and MELD score (OR=1.442, 95%CI: 1.225-1698, P＜0.001) were independent predictive factors for the prognosis of HBV-ACLF patients. MELD score combined with PWR had a stronger predictive efficiency than MELD score alone in predicting the prognosis of HBV-ACLF patients ［0.895 (95%CI: 0.827-0943) vs 0.842 (95%CI: 0.765-0.902), P＜0.05］. ConclusionMELD score combined with PWR can improve the efficiency of MELD score alone in predicting the prognosis of HBV-ACLF patients.