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Objective To explore the level of interleukin 35-producing B cells (i35-Breg) as well as its effect factors,interleukin-35 (IL-35),in peripheral blood of patients with chronic hepatitis B (CHB),and their relationship with hepatitis B virus (HBV) DNA and liver inflammatory degree.Methods A total of 35 treatment-naive CHB patients,17 interferon (IFN)-treated HBeAg-positive CHB patients and 15 healthy controls (HC) were enrolled.The levels of i35-Breg and IL-35 in peripheral blood were tested by flow cytometry and enzyme-linked immunosorption assay (ELISA).Kruskal-Wallis test,Wilcox rank sum test and two variables correlation analysis were used for statistical analysis.Results The percentage of i35-Breg cells as well as IL-35 level in peripheral blood of naive CHB patients were 3.05％ (0.89％,4.97％) and 2.81 μg/L (0.30 μg/L,12.33 μg/L),respectively,which were both significantly higher than those in HC group,which were 0.17％ (0.13％,0.45％) and 0.17 μg/L(0,1.93 μg/L),respectively.The difference were statistical significant (Z=-3.309 and-2.419,respectively,P=0.001 and 0.016,respectively).The peripheral level of i35-Breg was negatively correlated with the viral load in treatment-naive CHB patients (r=-0.529,P=0.008),while there was no correlation between the peripheral level of IL-35 and the viral load in treatment-naive CHB patients (r=0.11,P=0.54).The levels of i35-Breg and IL-35 in HBeAg positive CHB patients were 3.16％ (1.34％,5.62％) and 4.58μg/L (0.79μg/L,22.37 μg/L),respectively,which were both higher than those in HC group.The difference was statistically significant (F=3.39 and 3.37,respectively,both P＜0.01).Compared to HC group,the IL-35 levels in peripheral blood of CHB patients with ALT and AST levels less than 300 U/L were 3.03 μg/L (0.74 μg/L,22.37 μg/L) and 3.25 μg/L (0.83 μg/L,22.35 μg/L),respectively,with statistically significant difference (F=2.868 and 3.114,respectively,both P＜0.01).Compared to HC group,the peripheral level of i35-Breg in treatment-naive CHB patients with ALT levels less than 300 U/L was 3.14％ (1.03％,4.65％),with statistically significant difference (F=3.219,P=0.004).The IL-35 level showed a decreased trend in CHB who received IFN therapy,but there was no statistically significant difference (x2 =1.45,P =0.48).Furthermore,the baseline IL-35 level in patients who developed sustained viral response (SVR) was 0 (0,13.33 g/L),which was lower than that in patients who developed partial or primary no response 0.61 μg/L (0,24.72 μg/L).However,there was no statistical difference (F=0.75,P =0.68).Conclusions i35-Breg as well as its effect factor,IL-35,are involved in the progression of chronic HBV infection.The percentage of i35-Breg cells as well as IL-35 level in peripheral blood of treatment-naive CHB patients are increased.The peripheral level of i35-Breg is negatively correlated with the viral load,while there is no correlation between the peripheral level of IL-35 and the viral load.The percentage of i35-Breg cells as well as IL-35 level in CHB patients with low inflammatory degree are increased.
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Objective To investigate the efficacy and safety of different optimal therapy strategies for hepatits B e antigen (HBeAg) positive chronic hepatitis B (CHB) patients with suboptimal response to peginterferon-α-2a (peg-IFN-α-2a) at 24 weeks.Methods This open-label,single-center and prospective clinical observational study was conducted in Department of Infectious Diseases at Shanghai Changhai Hospital between January 2009 and December 2011.The cases of HBeAg-positive CHB with suboptimal response to peg-IFN-α-2a at week 24 were enrolled.Based on virological markers and patient preference,patients were treated with either peg-IFN-α-2a add-on adefovir dipivoxil (ADV) or switch-to telbivudine (LdT).Hepatitis B virus (HBV) virological and serological data were collected at week 12,24 and 48 after the initiation of optimal therapy.Adverse reactions were also monitored.Therapeutic efficacy was compared between two groups of patients before and after treatment by x2 test.Kruskall Wallis test and Mann-Whitney test were used for analysis of continuous variables.Results Among 193 HBeAg positive CHB patients treated with interferon,67 had suboptimal response and were enrolled.Forty five cases received peg IFN-α-2a add-on ADV treatment and 22 cases received switch-to LdT treatment.After 48 weeks of optimized therapy,the total tBeAg seroconversion rate was 25.3 ％.The rates of HBeAg loss,HBV DNA negative and alanine aminotransferase normalization were 26.8％,73.1％ and 83.5％,respectively.The peg-IFN-α-2a switch-to LdT strategy had better HBV DNA inhibition efficiency compared with the peg-IFN-α-2a add-on ADV strategy at week 12,24 and 48 (P=0.00,0.00 and 0.01,respectively).However,there was no significant difference of HBV DNA negative rate between two groups at week 48 (x2 =0.01,P=0.89).The obviously intolerable adverse reaction was not reported in two optimized strategy groups.Conclusions The 48-week optimized treatment for HBeAg positive CHB with suboptimal response to peg-IFN-α-2a at week 24 could achieve a higher HBeAg seroconversion rate.The switch-to LdT strategy may have better HBV DNA inhibition efficiency.Both strategies show satisfactory safety and tolerance.
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Objective To investigate expressions of programmed cell death ligand 1 (PD-L1) in hepatic tissues at the different stages of hepatitis B virus ( HBV) infection, and clarify its role in the mechanism of chronic hepatitis B virus infection. Methods The expressions of PD-L1 were detected by immunohistochemistry and computer image quantitative analysis in the hepatic tissues of 65 chronic HBV infected patients and 5 healthy controls. The correlations between PD-L1 expression and inflammatory grading in the hepatic tissues, total bilirubin (TBil), alanine aminotransferase (ALT), aspartate aminotransferase (AST), serum HBV DNA level were analyzed. Results The PD-L1 expressions in hepatic tissues of HBV infection with G0 - G4 inflammatory grades were 3. 07 % ±0.93%, 8.01%±1.49%, 11.60%±2.60%, 18.41%±2.21% and 26. 04% ±3. 41%, respectively,which were all significantly stronger than that in controls (0. 64%±0. 28%). PD-L1 expression was a positively correlated with inflammation grading of hepatitis tissues, TBil, ALT and AST level in serum (r=0. 917, 0. 787, 0. 483, 0. 628; all P<0. 05), and negatively correlated with serum HBV DNA load (r=-0. 620, P<0. 05). Conclusion The upregulated PD-L1 expression may be probably involved in the chronicity of HBV infection.
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ObjectiveTo investigate the effects of the imbalance between regulatory T cells (Treg) and T helper 17 cells (Th17) in patients with chronic hepatitis B virus (HBV) infection.MethodsThe serum concentration of Treg/Th17 differentiation-related cytokines in 34 patients with chronic hepatitis B (CHB),20 patients with HBV related acute on chronic liver failure (ACHBLF),and 20 healthy controls (NC) were measured by enzyme-linked immunosorbent assay (ELISA) and proportion of peripheral Th17 and Treg cells were analyzed by flow cytometry.Numeration data was analyzed by Fisher's exact propability method and measurement data was tested by one-factor analysis of variance or Turkey multiple comparison.Results The levels of Th17 differentiation-related cytokines,II-1β (3.97±2.85) pg/mL,IL-6 (12.75±-8.87) pg/mL,and IL-21 (360.0±335.7) pg/ mL in patients with ACHBLF were significantly increased than those in NC,which were (1.87 ±0.94) pg/mL(q=4.559,P＜0.01),(5.28±0.72) pg/mL(q=7.309,P＜0.01) and (46.68±20.17) pg/mL(q=6.946,P＜0.01 ),respectively.The proportion of Th17 increased markedly in patients with ACHBLF than that in NC(q=3.972,P＜0.05).However,compared to NC and patients with ACHBLF,the Treg differentiation-related cytokine,TGF-β,in patients with CHB,increased significantly (q=4.536 and 5.323,respectively; both P＜0.01).And the population of Treg also increased markedly in CHB patients.The level of IL-17A which was the characteristic effector cytokine of Th17 was the highest in patients with ACHBLF.The peripheral Th17 cell proportion was positively correlated with the level of serum total bilirubin in patients with ACHBLF (γ=0.74,P＜0.01).Conclusions Th17 and Treg imbalance including cytokine profiles and cell numbers exists in patients with chronic HBV infection.The Th17 are active in patients with ACHBLF and Treg are active in patients with CHB.
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Objective To identify the relationship between viral factors and disease progression in patients with acute hepatitis B virus (HBV) infection. Methods Ninety-seven adult patients with acute HBV infection in Shanghai Changhai Hospital were enrolled in this study and followed up for 24 weeks. Epidemiological, biochemical and virological parameters of all patients were collected. HBV S region from sera of 54 patients with acute HBV infection were genotyped using direct nucleotide sequencing. Differences of means between groups were compared by t-test, and frequency between groups was compared by X test. Results The clinical manifestations of all patients were mild and the 83 patients spontaneously developed HBeAg and HBsAg seroconversion. However, 14 patients had a tendency of chronicity, with HBV DNA level higher than patients without chronicity tendency [(6. 17 ±1. 04) 1g copy/mL vs (3. 86±1. 85)1g copy/mL;t = 5. 95, P<0. 01]. Among the 14 patients, 6 obtained HBsAg seroconversion after antiviral therapy and the other 8 developed to be sustained HBV carrier who had not received antiviral therapy. The main genotypes of acute HBV infection were genotypes B and C. There were no statistically significant differences of epidemiological factors and biochemical results between patients with the two genotypes of HBV infection. High viral load at baseline was the risk factor of chronicity tendency. Conclusions The main genotypes of acute HBVinfection in Changhai Hospital in the year from 2003 to 2007 are genotypes B and C. There is no significant relationship between genotype and clinical outcome. While high viral load at baseline is significantly associated with chronicity tendency. Proper antiviral therapy can decrease sustained HBV infection rate.
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Objective To investigate the distribution of genotypes in chronic HBV infection (CHB) and acute HBV infection (AHB) patients in Shanghai. Methods Sixty-two patients with AHB and 73 patients with CHB admitted to ('hanghai Hospital of Shanghai between 2003 and 2007 were studied. Viral genotypes of all the patients were determined by direct gene sequencing.Meanwhile, epidemiological, clinical and biochemical parameters of all patients were collected. Mean values of different groups were compared by t test while frequency was compared by chi square test. Results The major prevalent genotypes in both AHB and CHB patients were genotype B and C (48.4% vs 51.6% in AHB patients and 26.0% vs 74.0% in CHB patients). The proportion of genotype B was higher in AHB patients compared to CHB patients (P= 0.02). Epidemiological factors and clinical outcomes were not statistically different among patients with different viral genotypes. The proportion of genotype C was much higher in CHB patients compared to AHB patients (P=0.006). The main transmission route of AHB was heterosexual interaction which was 18 out of 62 (29.0%), but in CHB patients, it was prenatal transmission which was 38 out of 73 (52.1%). Conclusions In shanghai, the main HBV genotypes in both AHB and CHB patients are genotype B and C. The proportion of genotype B is relatively high in AHB patients while proportion of genotype C is more common in CHB patients. There is no significant relationship between genotypes and the clinical outcomes of AI-IB patients.
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Objective To study the inhibition of HCV IRES mediated HCV core protein expression in cells by inhibitor RNA. Methods Plasmid pcRz-IRNA, a eukaryotic expression vector with IRNA and two self cleavage ribozyme overhang at both sides respectively, was constructed and co-transfected with pcHCVcluc (containing HCV NCR, core and Luc genome) into the HHCC cell line (Human Hepatocellular Carcinoma cell line). Immunoflurescence tests were applied to detect the co-transfected cells, which were thereafter analysed with confocal microscope quantitatively. Luciferase activity was valued using Luc Assay System (Promega). Results The cotransfected cells expressed HCV core protein, and the fluorecein in which was reduced significantly in comparison with control. Conclusions IRNA can inhibit the expression of HCV IRES mediated core protein in the cotransfected cells.