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Objective: To determine the relationship between the serum hepatitis B virus (HBV) DNA and the risk of primary liver cancer (PLC)., Methods: Farmers aged 30 to 55 years in Long An county were recruited in this study Blood samples were collected and the sera were tested for HBsAg using Enzyme-Linked ImmunoSorbent Assay (ELISA), and the HBsAg-positive sera were further tested for viral DNA using nested polymerase chain reaction (nPCR). The study subjects were divided into three groups. The first group was positive for both HBsAg and HBV DNA. The second group was positive for HBsAg but negative for HBV DNA. Age-, sex-, residence-matched HBsAg negative controls for group 1 and group 2 were enrolled in the third group. The cohort was followed up for four years., Results: The positive rate of HBsAg in these farmers was 14.52% (3975/27,379), and the HBV DNA positive rate in HBsAg positive subjects was 40.35% (1604/3975). The total PLC incidence rate in Group 1 and 2 was 672.45 /100,000 person-years (PY), significantly higher than that in Group3 (17.19 /100,000 PY). The relative risk (RR) was 39.123, and the 95% confidence interval (CI) was 9.018-159.146. The PLC incidence rate of Group 1 (984.03/100,000 PY) was significantly higher than that of Group2 (324.38 /100,000 PY). The RR was 3.034, and the 95% CI was 1.795-5.125. Multivariate analyses of Group1 and 2 with Cox model showed that sex, age, serum HBV DNA, and family history of PLC were independent risk factors of PLC., Conclusion: HBV DNA and HBsAg positive subjects have a higher chance to develop PLC than HBV DNA negative-, HBsAg positive subjects.

Objective: To understand full-length sequence of HBV isolated from high incidence hepatocellular carcinoma area-Longan county, Guangxi., Methods: The nested polymerase chain reaction (nPCR) was used for amplifying the whole HBV DNA in sera of asymptomatic carriers. The products were sequenced by clone sequencing and homological analysis., Results: This isolate contained 3 215 bases. The genotype was C and the serotype was adw. There were 40 point mutations in polymerase gene which made 11 amino acids change. There were 11,2 and 3 point mutations in PreS1, PreS2 and S gene respectively which made 3,1,1, amino acids change. Six point mutations including the double mutations (nt 1762 A to T, 1764 G to A) were found in X gene leading to 4 amino acids change. There were 13 point mutations in C gene which made 2 amino acids change. No mutation was found in a determinant and Pre C. The isolate was quite close to the isolate from Vietnamese in evolution while far from the genotype C isolates from Shanghai, Beijing and Tibet., Conclusion: No special sequence was found in the isolate from high incidence hepatocellular carcinoma area, Longan county, Guangxi.

Objective: To explore the relationship between HBV core promoter mutations and liver damage or HBeAg status., Methods: Nested polymerase chain reaction (nPCR) was used for amplification of HBV DNA core promoter in 59 sera from patients with chronic hepatitis B in Guangxi, then the HBV DNA positive products were sequenced by direct sequencing., Results: The HBV DNA positive rate of was 59.3%(35/59). All the patients were infected by mutants. The commonest mutation was the double mutation (A --> T at nt1762 and G --> A at nt1764), counting for 57.1% (20/35). The next was C --> G at nt1799, counting for 54.4% (19/35), but this was no function. A --> G at nt1752 (resulting in isoleucine to valine) was seen in 37.1% (13/35) of the HBV DNA positive patients, and T --> C at nt1753 was seen in 20% (7/35). The significant difference in the frequency of T1762A1764 mutant was found between HBeAg positive patients (31.3%) and negative patients (79.0%)., Conclusions: HBV core promoter mutations are common among patients with chronic hepatitis B in Guangxi. T1762A1764 mutant is associated with HBeAg status and chronic hepatitis.

Objective: To understand the prevalence of HBV core promoter mutant (T1762 A1764 mutant) isolated from asymptomatic carriers from areas with higher and lower incidence of hepatocellular carcinoma (HCC) in Guangxi., Methods: A nested polymerase chain reaction (nPCR) was used for amplification of HBV DNA core promoter in sera, and then HBV DNA nPCR products were sequenced by direct sequencing., Results: The results show that 50.6% (39/77) of all HBV asymptomatic carriers were positive for HBV DNA HBV DNA positive rates of the samples from HCC higher incidence area, Longan County, and from lower incidence area, Guilin city were 55.6% (20/36) and 46.3% (19/41), respectively. HBV core promoter mutants could be seen in 35% in Longan positive samples and 47.4% in Guilin. The common mutations in both regions were all double mutations (nt 1,762 A-->T; nt 1,764 G-->A), accounting for 25% and 21%, respectively. The difference of the double mutant between Longan County and Guilin city was not significant (P>0.05)., Conclusions: These data implicated that the prevalence of HBV core promoter mutant isolated from asymptomatic carriers may not be correlated with the incidence of HCC in Guangxi.

Objective: To understand the distribution of hepatitis B virus genotype in Guangxi and its clinical significance., Methods: Nested polymerase chain reaction (nPCR) was used for amplification of HBV DNA in sera of asymptomatic carrier (ASC) of hepatitis B virus (HBV) and patients with different liver diseases from southern and northern Guangxi. Specimens from 161 subjects were positive for HBV DNA and HBV genotype was determined by using restriction fragment length polymorphism analysis, direct sequencing or cloning sequencing., Results: The prevalence of genotype A was 3.7% in all samples and that of genotype B, C and D was 21.7%, 72.7% and 1.2%, respectively. No other genotypes (such as genotype E, F, G, H) were found. The prevalence of genotype C showed an increasing trend in ASC, chronic hepatitis, liver cirrhosis and hepatocellular carcinoma (HCC) group; in contrast, the prevalence of genotype B showed an opposite trend, although no statistically significant difference was observed, except between ASC and HCC (P=0.05). The HBeAg positive rate was higher, and the anti-HBe positive rate was lower in patients with chronic genotype C infection than in those with genotype B (P<0.05 for both). Liver function test (ALT) abnormality was more severe in genotype C group than in genotypes A and B groups having acute or chronic infection (P<0.01 for all comparisons). The prevalence of genotype C in southern Guangxi was higher than that in northern Guangxi. In contrast, the prevalence of genotype B in southern Guangxi was lower than that in northern Guangxi., Conclusions: 1. The predominant HBV genotypes in Guangxi were genotypes B and C. The major genotype in southern Guangxi was genotype C; while that in northern Guangxi was genotype B, which implied that the distribution of HBV genotype C was consistent with the incidence of HCC in Guangxi. 2. Genotype C maybe associated with development of severe liver diseases including HCC. 3. Genotype A,D and B+C were mostly found in acute, hepatitis and chronic hepatitis group.