The Clinical Study Of Tpi Protein Expression On Clostridium Difficile In Diarrhea Patients Its Involved Mechanisms

Research background and objectives:In the last 30 years,the prevalence of Clostridium difficile infection?CDI?infections has steadily risen around the world and infection poses a global threat due to the emergence of antibiotic-resistant strains.Clostridium difficile is Gram-positive anaerobic bacillus.Clostridium difficile itself is not invasive,some toxigenic bacteria through the secretion of toxin A,toxin B and binary toxin cause antibiotic-related diarrhea,colitis and even fatal pseudomembranous enteritis,collectively referred to as Clostridium difficile infection?Clostridium difficile infection,CDI?¹,triosephosphate-isomerase is a C.difficile species-specific protein that is a housekeeping gene for C.difficile and a specific gene that distinguishes C.difficile from other bacteria.The tpi gene is highly specific and can be used for the rapid screening of Clostridium difficile in clinical samples ^2-3.tpi called triose phosphate isomerase is a glycolytic enzyme catalyzing the reaction between dihydroxyacetone phosphate and glyceraldehyde 3-phosphate.In Clostridium glycans,fatty acid synthesis,gluconeogenesis and pentose Metabolism plays an important role.The presence or absence of the triosephosphate-isomerase?tpi?gene was examined to determine whether C.difficile is present in the sample.Our research team found that outpatients and inpatient diarrhea patients have a higher detection rate of Clostridium difficile in our preliminary study.For the first time,we found a higher prevalence of Clostridium difficile infection in diarrhea patients with inflammatory bowel disease?IBD?.The incidence of tpi protein Is unique to Clostridium difficile,our group through the study of tpi protein expression of Clostridium difficile infection clear and IBD correlation,and then clear the distribution of tpi protein in IBD populations;the group also of C.difficile pathogenic The effects of bacterial tpi protein on diarrhea were studied.The effect of tpi protein of Clostridium difficile on the content of transferrin and the occult blood test OB was studied.Prepared to the experimental research on the mechanism of C.difficile in the development of diarrhea.Methods:1.Establishment of standard strain of Clostridium difficile for tpi protein purification and identification.2.312 in-patient stool samples were collected for incubation and purification of C.difficile and assessment of the epidemiological characteristics of Clostridium difficile and its toxin tpi protein load.3.The effects of Clostridium difficile and tpi protein on diarrhea were analyzed by statistics,and the influence of Clostridium difficile housekeeping gene tpi protein on transferrin and occult blood test?OB?was studied.4.To study the effect of tpi protein on adhesion,proliferation,apoptosis and angiogenesis in human normal colorectal mucosa?HIMEC colorectal mucosa cell line?.5.To investigate the effect of tpi protein on the expression of aquaporin 1?AQP1?in human intestinal microvascular endothelial cells?HIMECs?.Results:1,The low survival rate of C.difficile cultured in vitro was 24.3%,the detection rate of tpi protein in C.difficile was 85%.The positive rate of tpi protein in patients with diarrhea was significantly higher?16.9%?,the positive rate of tpi protein was significantly higher in IBD patients?19.3%?,tpi protein was significantly associated with diarrhea in IBD patients,and the risk of diarrhea in tpi patients was general diarrhea 2.13 times more patients?95%CI=2.340-8.123,P=0.0321?.2.The level of transferrin in patients with tpi protein positive was significantly higher than that in patients with positive tpi protein,tpi protein may be involved in the occurrence of IBD and promote the development of diarrhea in patients with IBD.3.Compared with other normal mucosal cell lines?GES-1 human gastric mucosal cells?HEEC normal esophageal epithelial cells,HOMF human oral mucosal fibroblasts?,tpi protein has a significant affinity for HIMEC cell lines,tpi protein inhibits HIMEC cell proliferation and promote apoptosis.4.The study of AQP1 on human intestinal microvascular endothelial cells?HIMEC?by tpi protein showed that the expression of AQP-1 protein and its mRNA and the uptake of HIMEC increased with the increase of tpi protein concentration in HIMEC?P<0.05?,suggesting that tpi protein can significantly inhibit the proliferation of intestinal mucosa?P<0.05?.Conclusions:1.Clostridium difficile infection is one of the causative factors leading to diarrhea.The tpi protein is associated with diarrhea,and the tpi protein may be a high risk susceptibility gene for diarrhea in IBD patients.2.Tpi protein may promote the progression of IBD patients.Tpi may promote the apoptosis of intestinal mucosal cells and reduce the intestinal water intake function,which may lead to diarrhea.