|
The background and purpose of Helicobacter pylori (Helicobacter pylori, H.pylori) is a spiral, microaerophilic Gram-negative bacilli colonization in the human gastric mucosa. 1983 Australian scholar Warren and Marshall isolated for the first time in human gastric biopsy tissue to bacteria. H.pylori chronic gastritis, the major causative agent of peptic ulcer, and gastric adenocarcinoma, gastric mucosa-associated lymphoid tissue lymphoma and primary gastric non-Hodgkin's lymphoma related. In 1994, the WHO and H.pylori classified as a class I carcinogenic factor. Around more than half of the population infected with H.pylori, Western countries H.pylori infection rates of 25-50%; developing countries, childhood H.pylori infection rate of about 60%, adulthood 80-90 %. Although H.pylori infection sustainable for life, but the majority of infected persons (80%) are asymptomatic carriers, only 10-20% of those infected chronic gastritis and peptic ulcer, a very small number of infected persons (0.1 -4%) may be the development of gastric cancer. Currently considered more important with the host genetic susceptibility and environmental factors related to H.pylori strain virulence differences on the severity of the clinical disease. Of H. pylori cag pathogenicity island (cag pathogenicity island, cag PAI) attracted much attention. Many epidemiological data cag of PAI-positive H.pylori infection and severe gastritis, peptic ulcer disease, atrophic gastritis and gastric cancer is closely related. Now clear that the cag of PAI encodes a type IV secretion system, the type IV secretion system has two functions: one CagA from H.pylori transported to the host cell; Second, induced gastric epithelial cells to synthesize and secrete cytokines such as IL -8. cag PAI the cagM, cagT well ORF13 protein expressed by the gene is considered to be an important structural component composed of type IV secretion system, missing the cagM, cagT or ORF13 mutant CagA can not be transported to the host cell, and can not be induced host cell synthesis and secretion of IL-8. ORF10 encoding the protein has nothing to do with the formation of the type IV secretion system, but as a guide CagA into the type Ⅳ sub 2004 pounds of bad of times palm master's degree thesis Xin secretion system channel missing ORF10 H Pylori mutant not transit CagA, but able to induce gastric epithelial cells to synthesize and secrete cytokines IL-8. As the and cagA, cagM, cagT, O stare 13 and ORF10 have important functions, and has certain distribution within cagl pair l Q distance and may be used as the testing complete cagPAI basis. To this end, the experiments were detected by PCR on cagl the cagA, cagM well cagH cagT to oRF 13, and oRF10 gene, in order to understand the high incidence of gastric cancer in Zhejiang Province of Daishan County the H Pylori cagI] AI distribution and analysis of each gene the gastroduodenal diseases types of relationships and the difference between [H Pylori, Hangzhou has ten. Materials and Methods accepted due to an upset stomach endoscopy in patients with gastric mucosal specimens H.Pylori isolated and cultured, 'Budai Shan County separation cultured H. Pylori124, strains, patients, male 90 cases, female 34 cases, age 16 71 years, with an average of 42.7 years of age, diagnosed by endoscopy and pathological examination of 60 cases of chronic gastritis, peptic ulcer, 59 cases, 5 cases of gastric adenocarcinoma; isolated and cultured in Hangzhou Hall Pylori47 strains, patients 25 males and 22 females, age 24-67-year-old Li \microaerophilic (5?, 10% cOZ, 85% N :) cultured for 5 days to select typical H Pylori colony growth, rapid urease test was positive, oxidase test positive, Gram staining Gram-negative form of was screw-shaped or birds like H Pyl, i, further pass substituting amplification bacteria cultured for 2 days. HPylori genomic DNA extracted using standard phenol / chloroform extraction method, according to the reported in the literature primers for PCR amplification, the reaction parameter is 95 ° C denaturation 3min, followed by 94OC305 56oC305 72oC455 of 30 cycles, and finally 72oC extend 7min. PCR products were subjected to electrophoresis using agarose gel electrophoresis detection the H. Pyzor Chuan strike Bed isolates of cagA eagM, eagT ORF13 and ORF10 positive experimental results using only test (SpSSll.5), where 0.05 was considered statistically significant. Results and Discussion H edge zori of cagA eagM eagT oRF13 and oRFlo positive rate: Dai 111 counties were 100.0%, 82.3%, 96.8%, 94.4% and 99.2%; Hangzhou were 100.0%, 93.6%, 97.9%, 957% and 100%, 82.3% chance of Daishan County cagA-positive strains with a complete cag of PAI the the complete cag team I in peptic ulcer, the 'net gastritis and gastric cancer patients detection rate were 84.7%, 80.0% and 80.0%, respectively; Hangzhou cagA-positive strains, 91.5% of the strains may complete cag PAI, complete cag PAI in the detection rate of peptic ulcer and chronic gastritis patients were 00.0%, 90.2% cag of PAI single gene in the digestive wins ulcers, 'of Long wins gastritis and gastric cancer detection rate: Daishan County cagA The detection rate of 100.0%; cagM 84.7%, 80.0% and 80.0%, respectively; cagT% .6%,% 66.7% 2004 Deficit j 'times palm master's degree thesis toot 100.0%; ORF13 94.9%, respectively, 95.0% and 80.0%; O 10 were 100.0%, 98.3% and 100.0% respectively. Hangzhou from peptic ulcer patients 6 H Pylori each gene detection rate was 100.0%; since slow separation of patients with gastritis H Pylori, positive rate of cagM's was 92.7%, 97.6% in cagT o good 13 to 95.1%, and ORF10 to 100.0%. cagA can not be used as a sign of the presence of a complete cag PAI; the cag pAI integrity and on detection of a single gene can be used to predict the type of the H Pylori stomach, duodenum disease; of dai 11) County compared with Hangzhou, Hall pylori eagA, eagM, eagT, O to stare of 13 and ORFlo-positive rate and there may be no significant statistical differences between the complete cag pAI of the ratio of H knife lori. Conclusions 11-I of the high incidence of gastric cancer in Zhejiang Province Dai County clinical isolates of H State ori strains, its the the cagA not as a complete cag pA] existing logo; cag The PAI's integrity and detection of single genes can not be used to predict the type of the H Pylori stomach, 11 duodenum disease; the HPylori from the high incidence of gastric cancer Daishan County?
|