| Hepatitis B, caused by hepatitis B virus(HBV), is the most common cause of chronic hepatic insufficiency, cirrhosis and hepatocellular carcinoma. According to the report from World Health Organization(WHO), more than 2.4 billion people are infected, of which over 780,000 people die every year due to the HBV-related disease. HBV infection is highly endemic in China with a positivity of 7.18% in the general population of hepatitis B surface antigen(HBs Ag). Therfore, it deduced that there are about 93 million HBs Ag-positive subjects in China, of whom about 20 million have chronic hepatitis B infection. Thus, HBV infection remains as one of the most important public health burdern in China. Recently, the relationship between diabetes mellitus and HBV infection has been paid more and more attention in epidemiological studies. Some studies supported the opinion of higher prevalence of HBV infection in diabetics than healthy controls, while the data from other researches suggested that no relationship between diabetes mellitus and HBV infection, while diabetes mellitus may play a role in the progression to liver cirrhosis or hepatocellular carcinoma in chronic HBs Ag carriers. In order to explore the characteristics of seroepidemiology and molecular epidemiology between diabetes mellitus and HBV infection, 3122 individuals were selected from “135” project of “Suzhou Industrial Park Centers for Disease Control and Prevention” for the present analysis and the main results are as follows. Part Ⅰ The correlation of seroepidemiology between diabetesmellitus and HBV infectionObjective:By analyzing the seroprevalence of the HBs Ag positivity and HBs Ab antibody positive in diabetics and non-diabetics individual, we investigated the relationships between diabetes mellitus and HBV infection.Subjects and Methods:3122 persons were recruited from “135” project of “Suzhou Industrial Park Centers for Disease Control and Prevention” in our study. Data on demographic characteristics, life styles, status of physical activity and history of diseases were obtained. Diagnosis of diabetes mellitus was based on the standard created by Chinese Diabetes Society. HBs Ag and HBs Ab were detected by enzyme-linked immuno sorbent assay(ELISA).The database was established and checked by Epidata3.0 software. All statistical analyses were conducted using SAS 9.2 software. An unpaired Student’s t-test or rank sum test was used for continuous variables. Differences in qualitative variables were compared by χ2. Multivariate analyses using logistic regression were performed to elucidate the relationship between HBs Ag seropositivity and diabetes mellitus. All tests were two-sided and differences were considered statistically significant if P values were <0.05.Results:1. Among 3122 participants, HBs Ag were detected positive in 514 persons, including 252 males(17.05%) and 262 females(15.94%)(P=0.402). 1447 subjects were seropositive for HBs Ab, including 685 males(46.35%) and 762 females(46.35%)(P=0.998).2. 365 of 3122 subjects were diabetics, among which 78(21.37%) persons were HBs Ag positive. HBs Ag were detected positive in 436(15.81%) people among 2757 non-diabetics. After the adjustment of age and gender, the difference of HBs Ag positivity between diabetics and non-diabetics was significant(OR: 1.47, 95%CI: 1.12-1.94).3. Compared to the persons in non-diabetics group, HBs Ag positivity was significantly higher in diabetics with BMI≥24kg/m2, waist circumference≥85cm(male) or 80cm(female), TC≥5.18(mg/d L), TG≥1.70(mg/d L), HDL-C≥1.04(mg/d L), LDL-C≥3.37(mg/d L).Conclusions:HBs Ag positivity was significantly higher in diabetics than those in non-diabetics. Among the diabetics who were overweight, obese, central obese, hypertensive or hyperlipidemia, the HBs Ag positivity was significantly higher.Part Ⅱ The correlation between mutations of HBV MHR region and diabetes mellitusObjective:To explore the distribution characteristics of HBV genotypes and to analyze the nucleotide mutations and amino acid substitutions in HBV MHR between diabetics and non-diabetics.Subjects and Methods:HBV DNA detection assays were completed in 514 HBs Ag positive persons, with 53 and 185 strains detected in diabetics and non-diabetics, respectively. 437 bp region of the surface gene was amplified by nested PCR. The amplification products were analyzed by gel electrophoresis on 1.5% agarose gel stained with ethidium bromide to determine HBV-DNA positive and negative samples. After purification with Gel-extraction kit, the PCR product was sequenced and analyzed by Meg Align v3.17 and MEGA 4.1 software.Statistical analyses were conducted using SAS 9.2 software. An unpaired Student’s t-test or rank sum test was used for continuous variables. Differences in qualitative variables were compared by χ2 or Fisher exact probability test. A two-sided P value of <0.05 was denote statistical significance.Results:1. Totally we obtained 238 HBV sequences, 53 from diabetics with 30 belong to genotype B and 23 belong to genotype C. 185 sequences from non-diabetics with 90 belong to genotype B and 95 belong to genotype C. There was no significant difference of the distribution characteristics of HBV genotypes between the strains isolated from diabetics and non-diabetics(P=0.307).2. 3 strains in diabetics contained stop codon mutations, among which one sequence showed a change from TGT to TGA at amino acid position 69 of HBs Ag. 2 strains got from diabetics showed a substitution at position 61 of HBs Ag, due to the TCA-TAA mutation. 4 strains in non-diabetics contained stop codon mutations, among which 3 sequence showed a substitution from TGT to TGA at amino acid position 69 of HBs Ag. One sequence of non-diabetics contained a stop codon at position 74(TGG-TGA).3. Totally, 49 strains were observed nucleotide substitutions in the 50 strains of diabetics in the MHR region, while 153 strains were observed nucleotide substitutions in non-diabetics(P=0.011); 86.00% of the sequences from diabetics and 70.17% from non-diabetics were detected nucleotide substitutions in “a” determinant region(P=0.025).4. Among genotype C HBV isolated from the present study, 50 strains from diabetics had a total of 25(2.05%) amino acids mutated in MHR, while 59(1.06%) substitutions were detected in 181 strains from non-diabetics(P=0.005).5. Increased amino acid mutations occurred within MHR in genotype C HBV isolated from diabetics more than 55 years old when compared to those from diabetics younger than 55 years old. Among genotype C HBV isolated from diabetics, more substitutions associated with vaccine escape were detected in the sequences from the persons more than 55 years old(P<0.05).Conclusions:1. There was no significant difference of the distribution of HBV genotypes between the strains from diabetics and non-diabetics.2. Increased nucleotide mutations were detected in the sequences from diabetics both in MHR and “a” determinant region.3. Increased amino acid mutations in MHR region were obseved in genotype C HBV-infected diabetics when compared with non-diabetics. Moreover, increased amino acid mutations in MHR region and more substitutions associated with vaccine escape were detected in the sequences from the persons more than 55 years old. |
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