Objective To study the relationship between liver histological changes and expression of HBcAg in peripheral blood dendritic cells in patients with chronic hepatitis B.Methods The peripheral blood samples were obtained from 24 patients with chronic hepatitis B (CHB) and 8 healthy individuals.The peripheral blood mononuclear cells (PBMCs) were isolated.DCs were isolated from peripheral blood mononuclear cells and enriched by culturing with recombinant human interleukin 4(rhIL-4) and recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF),then DCs were harvested and the costimulators of CD80,CD86,CD40 and CD54 on DCs were tested by immunofluorescence flow cytometry.These costimulators were compared with liver histological changes.Results Percentage of CD40 and CD86 was significant lower than that of healthy donors(P

Objective To assess the liver biopsy and the clinical characteristics of inactive HBsAg carriers.Methods One hundred and ten inactive HBsAg carriers,including 76 males and 34 females aged (38.9 ± 9.4) years (21-66),underwent liver biopsy from January 2011 to September 2015,the histopathological findings and clinical features were analyzed.Among 110 cases the inflammation activity (A) was ＜ A2 in 73 cases and ≥A2 in 37 cases;the fibrosis (F) ＜ F2 in 63 cases and ≥F2 in 47 cases.The upper limits of normal (ULN) for ALT was defined as 30 U/L for men and 19 U/L for women according to World Health Organization (WHO) standard,and 50 U/L for men and 40 U/L for women according to Chinese national standard.There were 59 cases with ALT ＜ 1 × ULN of WHO standard and 110 cases with ALT ＜ 1 × ULN of Chinese standard.Results In 110 inactive HBsAg carriers,there were 100 cases (90.9％) ≥A1 and 37 cases (33.6％) ≥A2,84 cases (76.3％) ≥F1 and 47 cases (42.7％) ≥F2.The severity of A and F were both higher in males than that in females,especially that of F (U =2.162,P =0.032;x2 =5.315,P =0.021).But there were no statistical differences between WHO standard group and Chinese standard group (U =0.951,0.435;P =0.341,0.663).Along with the increase of age,the degrees of A and F aggravated (F =3.705,5.915;P =0.014,0.001).The average ages in ≥ A2 group and ≥F2 group were (41.7 ± 9.6) years and (38.7 ± 8.1) years,respectively.The independent risk factors for severity of A and F were age,gender (male) and age,respectively.Conclusion There may be histological damages of varying degree in liver tissues of most inactive HBsAg carriers,and for those aged 40 years and over,especially males screening of liver histological activity and fibrosis would be necessary.

Objective To evaluate HBeAg quantification in predicting the efficacy of pegylated interferon (PegIFN) α treatment for patients with HBeAg-positive chronic hepatitis B (CHB).Methods A total of 216 HBeAg-positive CHB patients admitted in Ningbo No.2 Hospital during March 2009 and December 2011 were enrolled in the study.All patients were given subcutaneous injection of PegIFNα-2a or PegIFNα-2b weekly for 48 weeks and followed up for 24 weeks after discontinuation.Patients were divided into HBeAg seroconversion group and non-seroconversion group at the end of the follow-up.Receiver operating characteristic (ROC) curves were used to evaluate HBeAg levels at baseline and 12,24 weeks of treatment in predicting HBeAg seroconversion.Results HBeAg seroconversion was observed in 31.48％ (68/216) patients at the end of follow-up,and there was no significant difference in seroconversion rate between patients treated with PegIFNα-2a and those with PegIFNα-2b (32.00％ vs.29.27％,P ＞ 0.05).There was significant difference in baseline HBeAg levels between patients with HBeAg seroconversion and those without HBeAg seroconversion (Z =-3.834,P ＜ 0.05).HBeAg seroconversion patients had a tendency of rapidly decreasing HBeAg level,but there was no significant difference in decreasing rate between seroconversion and non-seroconversion patients (F =3.321,P ＞ 0.05).ROC curves showed that HBeAg level at 24-week was the best indicator for predicting HBeAg seroconversion with area under curve of 0.861.Conclusion Serum HBeAg level at 24-week of treatment may be used to predict the HBeAg seroconversion in HBeAg-positive CHB patients treated with PegIFNα.

Objective To investigate the efficacy of polyethylene glycol (PEG)-interferon α (PEG-IFNα) in treating HBeAg-positive chronic hepatitis B (CHB) and explore the relationship between hepatitis B virus (HBV) genotypes and the effect of interferon α (IFNα) therapy.Methods A total of 199 CHB patients with known genotypes were given subcutaneous injection of PEG-IFNα-2a or PEG-IFNαt-2b once a week for 48 weeks,with another 24 weeks follow up.The seroconversion of HBeAg influenced by HBV genotypes were analyzed after discontinuation of treatment.Results In local area,genotype C was the major genotype[64.32％ (128/199)].Except serum ALT and AST level,the differences in gender,age,liver inflammation,degree of liver fibrosis,HBeAg level and HBV DNA level between genotype B and C were not statistically significant(all P ＞0.05).The seroconversion rate of HBeAg in patients with genotype B at early stage of therapy (3 months) was significantly higher than that of patients with genotype C [26.76％ (19/71) vs 10.16％ (13/128),x2 =9.330,P =0.002].While at the end of follow-up,seroconversion rate of HBeAg in patients with genotype B (followed up for 6 months) was higher than that of patients with genotype C [39.44％ (28/71) vs 30.47％ (39/128)],but the difference was not statistically significant(x2 =1.645,P =0.200).By univariate analysis based on log-rank test,the time of HBeAg seroconversion in patients with genotype B was much earlier than that of genotype C [(13.99 ± 0.67) months vs (15.47 ± 0.41)months],but the difference was not statistically significant (P =0.150).Conclusions The seroconversion rate of HBeAg in patients with genotype B treated with PEG-IFNα was significantly higher than that of genotype C in early stage of therapy (3 months),while similar at the end of therapy.

Objective : To learn the characteristics of second-line drug resistance and related gene mutations of multidrug-resistant Mycobacterium tuberculosis ( MDR-TB ) Beijing genotype strains. Methods: The MDR-TB isolates in Hwa Mei Hospital from 2017 to 2019 were enrolled and detected using RD105 deletion-targeted multiplex polymerase chain reaction (PCR). The proportion method for drug susceptibility test was used to detect the drug-resistant profiles against kanamycin, amikacin, capreomycin, ofloxacin and levofloxacin. The gene sequencing of rrs, tlyA, eis, gidB, gyrA and gyrB was conducted by PCR compared with H37RV strain. The differences in the rates of drug resistance and mutation between Beijing and non-Beijing genotype strains were examined to understand the characteristics of Beijing genotype strains. Results: There were 106 Beijing genotype and 27 non-Beijing genotype strains in 133 MDR-TB isolates. The drug resistance rates of kanamycin, amikacin, capreomycin, ofloxacin and levofloxacin in Beijing genotype strains were 9.43%, 7.55%, 3.77%, 32.08% and 32.08%, respectively. The rates of quasi-extensive and extensive drug resistance in Beijing genotype strains were 30.19% and 7.55%. The gene mutation rates of rrs, tlyA, eis, gidB, gyrA and gyrB in Beijing genotype strains were 7.55%, 7.55%, 1.89%, 2.83%, 36.79% and 2.83%, respectively. There were no significantly differences between Beijing and Non-Beijing genotype strains in the factors above ( P>0.05 ). The gene rrs, tlyA, eis, gidB, gyrA and gyrB had 2, 1, 2, 2, 5 and 3 mutation types, respectively, with single base substitution as the main type. Conclusion Beijing genotype strains are dominant in MDR-TB, with high resistance to fluoroquinolones and mainly gyrA gene mutation.

Objective To investigate the intensity of HBsAg and HBcAg expression in liver tissue of patients with chronic hepatitis B virus (HBV) infection and its clinical significance.Methods A total of 994 HBV infected patients underwent liver biopsy and histopathological examination.The expression of HBsAg and HBcAg in liver tissue was detected by histoimmunochemistry.Patients were divided into HBeAg (+)/HBVDNA(+), HBeAg(-)/HBV DNA(+) and HBeAg(-)/HBV DNA(-) groups according to HBeAg and HBV DNA levels;patients were divided into ＜2 × normal (ULN) group, 2-＜5 × ULN groupand ≥5 × ULN group according to the alanine aminotransferase (ALT) levels.The histologic activity (A), fibrosis (F), the expression of HBsAg and HBcAg in liver tissue and their correlations with clinical features were analyzed.Logistic regression analysis was used to study the factors affecting the expression of HBsAg and HBcAg in liver tissue.Results Among 994 HBV infected patients, 941 cases (94.67％) were intrahepatic HBsAg positive and 553 cases (55.63％) were intrahepatic HBcAg positive;403 cases (40.85％) were ≥A2 in histologic activity and 371 cases (36.09％) were ≥F2 in fibrosis.The degree of A and F was the highest in HBeAg (-) / HBV DNA (+) group, followed by HBeAg (-) / HBV DNA (-) group, and was the lowest in HBeAg (+) / HBV DNA (+) group.The intensity of intrahepatic HBsAg expression was significantly different among three groups (x2 =6.299, r =-0.760, P ＜ 0.05), however, the difference was not showed in pairwise comparisons.The difference of intrahepatic HBcAg intensity among three groups was statistically significant (x2 =282.995, r =-0.645, P ＜ 0.01), the intensity was the highest in HBeAg (+) / HBV DNA (+) group and the lowest in HBeAg (-) / HBV DNA (-) group.The constituent ratio of HBeAg positive and HBV DNA level were higher and the average age was lower in intrahepatic HBsAg positive group than those in HBsAg negative group.The constituent ratio of positive HBeAg, the levels of ALT, AST, PLT and HBV DNA were higher and the average age, the average FIB-4 level were lower in intrahepatic HBcAg positive group than those in HBcAg negative group.The HBV DNA level was an independent risk factor for intrahepatic HBsAg intensity, and the HBeAg positive and HBV DNA level were independent risk factors for intrahepatic HBcAg intensity.There were no significant differences in A and F among different groups of intrahepatic HBsAg intensity (x2 =1.943 and 2.630, both P ＞ 0.05).There was significant difference in F among different groups of intrahepatic HBcAg intensity (x2 =12.352, P ＜ 0.01), but not in A.The degree of F was the highest in intrahepatic HBcAg negative group.There was significant difference in intrahepatic HBcAg intensity among different groups of ALT level (x2 =16.349, P ＜ 0.01), but not in intrahepatic HBsAg intensity.The intrahepatic HBcAg intensity in ALT ＜ 2 × ULN group was lower than that in other two groups.Conclusions Most of patients with chronic HBV infection are intrahepatic HBsAg positive and more than half of them are intrahepatic HBcAg positive.The intrahepatic HBsAg intensity is not associated with A and F, but correlates with HBV DNA level.The intrahepatic HBcAg intensity is not associated with A, but it is negatively correlated with F and positively correlated with positive HBeAg expression, HBV DNA level and ALT level.

Objective To analyze the correlation between liver pathology and clinical characteristics in a large series of patients with chronic HBV infections , so as to provide the data base for non-invasive medical diagnosis .Methods Liver pathology and clinical characteristics of 1 397 patients with chronic HBV infections were retrospectively analyzed . Ridit analysis and Spearman correlation analysis were performed to investigate the correlations of clinical characteristics with liver pathology of patients .Results In 1 397 patients, there were 604 patients (43.24%) with liver inflammation grading ≥G2 and 504 patients (36.08%) with fibrosis stage ≥S2.Inflammation grade and fibrosis stage of liver tissues were both higher in male patients than those in females (u=3.093 and 2.854, P<0.01).Inflammation grade and fibrosis stage of liver tissues in patients aged ≤30 years were lower than those in patients aged >30-40 years and >40 years (r=0.259 and 0.303, P<0.01;F=4.199 and 12.226,11.610 and 24.359, P<0.05 and <0.01).Patients with HBeAg( -) and HBV DNA≥103 copies/mL were of higher degrees in liver tissue inflammation compared with those with HBeAg ( +) and those with HBeAg ( -) but HBV DNA <103 copies/mL (F=8.788 and 5.635, all P<0.01);while the fibrosis stage in patients with HBeAg (-) and HBV DNA≥103 copies/mL was only higher than that in HBeAg ( +) patients (F=12.886, P<0.01). Liver tissue inflammation and liver fibrosis aggravated with the increase of ALT ( r=0.537 and 0.517, P<0.01).There was no significant difference in liver tissue inflammation among different age groups of patients with ALT (1-<2) ×ULN and HBV DNA≥103 copies/mL (χ2 =4.365, P >0.05),but there was significant difference in liver fibrosis in patients between aged >40 years and ≤30 years ( F=3.177,P<0.05).Conclusions Liver biopsy and antiviral therapy should be considered in chronic HBV infected patients with age of >30 years, lightly elevated ALT levels , HBeAg(-) and detectable HBV DNA levels , especially in male patients .Screening for liver fibrosis should be considered in patients with HBeAg ( -) and low HBV DNA levels .

OBJECTIVETo explore the effect of silencing the Notch2 gene by small interfering (si)RNA on the proliferation of the HepG2 human hepatocellular carcinoma (HCC) cells.

METHODSNotch2-siRNA was transfected as a liposomal formulation into HepG2 cells. The non-HCC cell lines SG07901 (gastric cancer) and SW620 (colon cancer) were used as controls. The mRNA expression of Notch2 and Hesl were detected by RTPCR, and the protein expression of Notch2 was detected by western blotting. The proliferation of transfected HepG2 cells was assessed by the cell counting kit-8 (CCK8) colorimetric assay.

RESULTSThe untransfected HepG2 cells showed significantly upregulated transcript expression of Notch2, and not of Notch1, Notch3 or Notch4, compared to the other non-HCC cell lines. Following transfection of Noteh2-siRNA into HepG2 cells, the mRNA expression of Notch2 and Hes1 and the protein expression of Notch2 were significantly decreased. The rales of proliferation inhibition in HepG2 following transfection of Notch2-siRNA showed an increasing time-related trend, with 2.64% ± 1620% at 12 h, 38.34% ± 8.80% at 24 h, 70.05% ± 7.80% at 48 h, 70.78% ± 10.00% at 72 h, and 74.22% ± 4.80% at 96 h.The inhibition rate at 24 h of transfection was significantly different from that of the groups of control cells.

CONCLUSIONNotch2 is upregulated in the common HCC cultured cell line HepG2. siRNA-mediated silencing of Notch2 exerts inhibition effects on HepG2 proliferation, suggesting the potential for this approach as targeted therapy for treating HCC.

Carcinoma, Hepatocellular ; pathology ; Cell Proliferation ; Down-Regulation ; Hep G2 Cells ; Humans ; Liver Neoplasms ; pathology ; RNA Interference ; RNA, Small Interfering ; Receptor, Notch2 ; metabolism

Objective To comparatively analyze the immunological characteristics of patients with mild and severe influenza A (H1N1), and to provide the evidence for condition monitoring and treatment . Methods 52 cases with mild influenza A ( H1N1), 152 cases with severe influenza A ( H1N1) and 26 healthy subjects from July 1, 2009 to December 31, 2009 were enrolled in the study.Lymphocyte subsets in peripheral blood were analyzed by flow cytometry and the serum concentrations of interferon -γ( IFN-γ) and interleukin-4 (IL-4) were detected by enzyme-linked immune-sorbent assay (ELISA).Results The total lymphocyte counts were decreased obviously in patients with severe influenza A ( H1N1) than in mild pa-tients and in healthy subjects (P<0.01).The T lymphocyte, NK cells, CD4+T lymphocyte and CD8+T lym-phocyte were also decreased obviously in severe patients than in mild patients (P<0.01).The B lymphocyte and CD4+/CD8+were also decreased in severe patients than in mild patients but had no significant statistical difference (P=0.11, 0.175).The serum IFN-γlevels in patients with mild and severe influenza A (H1N1) were lower than those in control group, especially in patients with severe influenza A (H1N1) (compared with control group and mild group , P<0.01).And the changes of serum IL-4 levels were the same with the former, but there were no statistically significant differences in three groups (P>0.05).Con-clusion Immune dysfunction in patients with influenza A (H1N1) infection is associated with the severity of disease, especially cellular immunity .Therefore, monitoring of the immune system is valuable for the diag-nosis of influenza A(H1N1) infection.

Objective: To analyze the distribution of HCV genotype in eastern Zhejiang Province and its correlation with sex, age, viral load, antiviral effect and so on. Methods: A total of 501 cases of HCV infection seen in Ningbo No. 2 hospital from January 2011 to April 2018 were included. The HCV genotypes and HCV RNA were detected by gene chip method and RT-PCR respectively. The liver function and blood routine tests were performed and the APRI index was calculated. The factors affecting the SVR were analyzed for the patients who were partially treated with pegylated interferon and ribavirin (PR). Results: The HCV genotypes of 501 cases were 1b、6、2a、3a、3b、1a from the higher to lower ranks, and genotype 1b was more than 50%.The distribution of HCV genotypes in different age groups was significantly different (χ²=95.433, P<0.001). The age of patients with 1a, 1b and 2 A was significantly higher than that of 3a, 3b, and type 6 (F=11.879, P<0.001). There were significant differences in viral load, AST, PLT and APRI index among different genotypes (F=2.920, χ²=12.400, F=6.294, χ²=12.700, all P<0.013), but ALT had little difference (χ²=7.994, P>0.157); 179 patients with PR standard regimen were followed up. The result showed that there was no significant difference in the overall efficacy of different genotypes (χ²=6.598, P=0.252), but the rate of sustained virological response in type 3 A was significantly higher than that of the 1b type (χ²=4.193, P=0.041). Conclusions The HCV genotypes are mainly 1b and 6 in eastern Zhejiang Province. There were significant differences in age, viral load, AST, PLT and APRI indices among patients with different HCV genotypes, and the therapeutic effect of PR regimen was better for genotype 1b than that of 3a.