Objective To investigate the clinical significance of HBV pre-core and basal core promoter mutations in chronic hepatitis B using Gene-chip technique.Methods Four spot mutations of 1896,1814 dots in pre-core region and 1762,1764 dots in basal core promoter region were detected in 95 patients with chronic hepatitis B using gene chips.Serum HBV DNA level was measured using fluorescent quantitation polymerase chain reaction(FQ-PCR).Results In the 95 patients with chronic hepatitis B, HBV pre-core or basal core promoter mutations were detected in 91 patients,and the positive rate was 95.8%.The mutation rate of G1896A,A1762T associated with G1764A,and G1896A plus A1762T plus G1764A were 36.3%,70.3% and 24.2% respectively.No A1814C mutant was detected in these patients.Compared with no mutation group,the serum HBV DNA quantity was lower in G1896A plus A1762T and G1764A mutation group,and no significant changes was found in other groups.Conclusion Several different variant sites of HBV can be detected at one time using gene chip technique.HBV pre-core and basal core promoter mutations have effect on the serum HBV DNA level in chronic hepatitis B patients to a certain extent.

Objective To investigate hepatitis B virus (HBV) gene mutations related to entecavir (ETV)-resistance in patients with chronic HBV infections.Methods Serum samples were collected from 44 patients with chronic HBV infections and resistant to ETV treatment who were admitted in Ningbo No.2 Hospital during February 2010 and May 2014.The HBV polymerase regions were amplified by real-time fluorescent quantitative polymerase chain reaction (PCR) method,and the PCR products were analyzed with direct sequencing.SPSS 16.0 was used to assess the frequency of HBV polymerase gene mutations,and its relation to the viral genotype and clinical features.Results The most common HBV polymerase gene mutation was rtS202G/I (52.28％,23/44),followed by rtT184A/G/I/S (36.36％,16/44) and rtM250V/L (11.36％,5/44).Nine mutation patterns were detected,in which rtL180 + rtM204V + rtS202G/I (38.64％,17/44) and rtL180 +rtM204V + rtT184A/G/I/S (27.27％,12/44) were the most frequent ones.The difference in gene mutations between genotype B and C was of statistical significance (x2=12.294,P ＜0.01).Patients carrying rtT184A/G/I/S mutations were associated with worse liver function (x2 =14.499,P ＜ 0.01),and those carrying rtM250V/L mutations were associated with lower HBeAg positive rate (x2 =10.057,P ＜ 0.01).Conclusions rtL180M + rtM204V + rtS202G/I is the most common HBV polymerase gene mutation related to ETV resistance in patients with chronic HBV infections.Different gene mutations may be associated with HBV genotypes,severity of liver damages,and HBeAg positive rate.

Objective To evaluate the clinical application of liver shear wave velocity (SWV) in comparison with common serum score systems [AST to platelet ratio index (APRI),Forns,S index and FIB-4] in assessment of liver fibrosis in patients with chronic hepatitis.Methods A total of 237 chronic hepatitis patients with liver fibrosis confirmed by liver biopsy,who were admitted in Ningbo Second Hospital during October 2010 and April 2013,were enrolled in the study.Liver shear wave velocity were measured by acoustic radiation force impulse (ARFI),and the score of APRI,Forns,S index and FIB-4 were calculated based on the measurement of serum markers.Liver fibrosis stages were classified as S0-S4 according to the Scheuer scoring system,and stages ≥ S2 were identified as significant liver fibrosis.The diagnosis value of SWV,4 common score systems and their combination for significant liver fibrosis was evaluated by receiver operating characteristic curve (ROC).Results A significant linear correlation was found between SWV and the stage of fibrosis (r =0.46,P ＜ 0.01).The areas under the ROCs of SWV and 4 common score systems (APRI,Forns,S index and FIB-4) for the diagnosis of significant liver fibrosis were 0.758 (0.696-0.821),0.727 (0.662-0.793),0.777 (0.717-0.836),0.747 (0.684-0.810) and 0.737 (0.673-0.802),respectively.The area under the ROC of the combined prediction nodel established with Logistic regression was 0.810.Conclusion Liver shear wave velocity measured by ARFI is of clinical value in noninvasive assessment of liver fibrosis,and the prediction accuracy can be improved when it is combined with other noninvasive indices.

Objective To investigate the relationship between HBV polymerase gene mutations and HBV genotypes in chronic hepatitis B (CHB) patients resistant to adefovir dipivoxil (ADV).Methods Blood samples were collected from 114 ADV-resistant CHB patients during February 2010 and May 2012.The HBV polymerase regions from serum samples were amplified with real-time PCR,and the PCR products were sequenced.Normal distribution data were presented as x ± s,and non-normal distribution data were presented as M (P25-P75) ; for homogeneous data analysis of variance and LSD-t test were performed.Results There were 8 types of HBV polymerase gene mutations in 114 CHB patients; single point mutation was detected in 102 patients (89.47％) and double or triple points mutations were detected in 12 patients (10.53％).rtA181V/T/S (57.89％),rtN236T (14.91％) and rtA181V/T/S + N236T (9.65％) were the predominant mutations.For 21 patients (18.42％) with HBV genotype B,rtN236T mutation was prevalent (47.62％,10/21) ; while for those with HBV genotype C (93,81.58％),rtA181V/T/S mutation was the predominant (65.59％,61/93).The differences of rtA181V/T/S (x2 =12.269,P ＜0.01) and rtN236T (x2 =18.658,P ＜0.01) mutation rates between B and C genotypes were statistically significant.Conclusion rtA181V/T/S,rtN236T and rtA181V/T/S + rtN236T are the major HBV polymerase gene mutation types in ADV resistant CHB patients,and mutation types are related with HBV genotypes.