Identification Of Helicobacter Hepaticus In Various Strains Mice From Different Regions Of China And Helicobacter Pylori Infection In Patients With The Chronic Hepatitis B

Part 1 Identification of Helicobacter hepaticus infection in various strains mice from different regions of ChinaAim: To identity the prevalence of H.hepaticus infection in various strains mice from commercial and academic institutions in different regions of China.Methods: We tested the prevalence of serum anti-H.hepaticus-IgG and feces H.hepaticus antigen in 102 various strains mice from different regions of China. The feces received from all mice were srceened for Helicobacter genus-specific 16SrRNA and H.hepaticus species-specific 16SrRNA by PCR and were cultured and identitied for Helicobacter species. With above methods, we could analyze the H.hepaticus infection.Results: 23 (22.55%) mice were infected with Helicobacter species in 102 mice and 51.17% (12/23) were H.hepaticus infection. Among the 39 mice from the laboratory animal center of Jilin University, 8 mice were infected with Helicobacter species and 50.00% (4/8) were H.hepaticus infection, and the 4 mice were from the same facility. Among the 26 mice from the laboratory animal center of Nanjing, 14 mice were infected with Helicobacter species and 53.33% (8/15) were H.hepaticus infection, and the 8 mice were from this center and an affiliated facility of this center. Among the 12 mice from t the laboratory animal center of Shanghai, only 1 mouse was infected with Helicobacter species and it was from an affiliated facility of this center; but none of mice infected H.hepaticus was found. Among the 25 mice from the laboratory animal center of Beijing, none was infected with Helicobacter species. H.hepaticus was the main infected strain in laboratory mice and A/J mice were the susceptible strains. At the same time, the other positive PCR amplicons of Helicobacter genus-specific 16SrRNA with H.hepaticus species-specific 16SrRNA negative were sequenced and it was found that other Helicobacter species could infect the laboratory mice.Conclutions: From the laboratory animal center of Jilin University, Nanjing and Shanghai, Helicobacter species and H.hepaticus could infect various strains mice. The prevalence of the laboratory animal center of Nanjing was the highest, then was the laboratory animal center of Jilin University, and no findings was in the laboratory animal center of Beijing. H.hepaticus was the main strain infected mice, but also there might be exist other Helicobacter species infected mice. The PCRmethod from feces was more sensitive than the examination of the antigen of H.hepaticus from feces and the antibody of H.hepaticus from serum.Part 2 Seroprevalence of H.pylori infection in patients with the chronic hepatitis B and the association with the virus loads and genotypes of HBV DNAAims: To investigate the seroprevalence of H.pylori infection in patients with the chronic hepatitis B and the synergistic effect with HBV on the development of the chronic hepatitis B.Methods: In this case-control study, the cases were 502 patients with the chronic hepatitis B (357 males, 145 females, mean age 34.2±13.2 years). The controls were 429 sex and age matched healthy donors (322 males, 107 females, mean age 33.6±14.1 years) and 396 patients with chronic gastrititis (199 males, 97 females, mean age 33.8±13.3 years). All subjects were tested for presence in serum of IgG antibodies against H.pylori, and the cases were tested the quantitation and genotypes of HBV DNA. The result was analyzed using the chi-square test.Results: H.pylori infection was more prevalent in patients with the the chronic hepatitis B (59.5%), the chronic HBV-related cirrhosis (77.1%), and the HCC (80.6%) than in healthy controls (43.4%)(p<0.05). Moreover, the seroprevalence of H.pylori in patients with the chronic cirrhosis and the HCC was higher than that in patients with the chronic hepatitis (p<0.05). H.pylori infection aggravated with the progress of the disease. With the different virus load of HBV DNA, H pylori prevalence all increased, but there was no significant difference among the groups of the different virus loads. H pylori prevalence in patients with genotyping A ,B,C and D was 33.3%,51.6%,50.0% and 54.5%, respectively. There was no significant difference among the genotypes. H.pylori infection was prevalent obviously in cirrhosis patients with complication such as hepatic encephalopaphy, peptic ulcer and upper gastrointestinal hemorrhage than that in patients without complications.Conclusion: H.pylori seroprevalence was higher in patients with chronic hepatitis B than in heathy controls. H pylori might play the synergistic effect with HBV on the development from the chronic hepatitis B to the cirrhosis and the hepatocellular carcinoma.Part 3 Identification of Helicobacter genus-specific 16SrRNA and H.pylori cagA, vacA and glmM genes in human liver samples from patients with the chonic hepatitis BAims:Several studies have shown the presence of Helicobacter species in the human lilver samples of the chronic hepatitis C and hepatocellular carcinoma (HCC) and in the intestinal tract of animals. Experimental infection by H.hepaticus in mice causes the chronic hepatitis and HCC. This study investigated whether Helicobacter genus-specific 16SrRNA could be detected in the liver samples from patients with the chronic hepatitis B.Methods:Liver samples from 56 patients with the chronic hepatitis B diagnosed by histopathology were studied. Histology with standard culture and polymerase chain reaction (PCR) amplification was performed with Helicobacter genus-specific 16SrRNA primers and the specific primers of H.pylori cagA,vacA and glmM genes to detect the presence of bacteria. Positive amplicons were identified by sequencing.Results:Helicobacter genus-specific 16SrRNA was found in 35 of 56 samples of patients with the chronic hepatitis B. We further performed PCR with the specific primers of H.pylori cagA,vacA and glmM genes and the cagA, vacA or glmM genes were detected in 21 samples of 35. The other positive amplicons with only Helicobacter genus-specific 16SrRNA were sequenced and were found to have 99.2% similarity to the 16SrRNA of H.hepaticus and 98.7% similarity to the 16SrRNA of H.rodentium, 96.9% similarity to the 16SrRNA of H. pullorum, 95.2% similarity to the 16SrRNA of H.bilis.Conclusions: Helicobacter DNA can be present in the liver of patients with chronic hepatitis B and is probably linked to progess from hepatitis to cirrhosis and HCC in the liver.