16S RDNA-base Analysis Of Bacterial Diversity In Th Microbial Flora Of Renal Transplant Patients Intestinal Tract

Objective:Intestinal bacteria mediated metabolism,immunity,infection and other aspects,and was closely related to human health.All kinds of drugs used in renal transplantation and system state change was bound to impact on the intestinal flora,which may lead to increased risk related diseases.The purpose of this study was to investigate the effect of renal transplantation on intestinal flora and the possible risks in patients with renal transplantation.Methods:Part I:Collected stool samples from patients which received renal transplantion in the PLA 309 hospital urology from September 2015 to October,within 3 days before surgery and on postoperative day 28.We used self control design,based on 16 S rDNA high-throughput gene sequencing technology,and Silva database,softwares of mothur,graphpad,R language,LEfSE was used to analyze the microbial diversity.Part II: Further analysis of flora structure which patients used sulfa drugs long-term,Fecal samples were collected,within 3days before surgery and on postoperative day 28,from patients(10 cases)which received renal transplantion in the PLA 309 hospital urology from June 2016 to August.Experimental method was same as part I and further compared two different states' effects on intestinal flora.Results:1.In the two parts of the experiment,The pyrosequencing separately provided 402857(14 samples)and 737730 reads(20 samples)in total.In both experiments,Rarefaction curve and Shannon-Wiener of all samples were basically flat with the increase of the depth of sequencing,indicating that the amount of sequencing data was enough.2.The measured OTU,Ace,Chao,Shannon,Simpson and other index were used to assess the?-diversity of the intestinal flora and found that the first part of the experiment besides Simpson index increased,other indicators showed a downward trend but no significant difference,while the Simpson index increased significantly in the second part of the experiment,the other indexes were all decreased significantly.3.The principal coordinate analysis(PCoA)and the similarity dendrogram were used to evaluated intestinal flora ?-diversity,PCoA showed,in the first part of the experiment,some samples were overlaid and the two groups before and after operation can barely distinguish.But in the second part of the experiment,although there were a few samples mixed,two groups divided into two.The result of similarity tree was similar.In the two part of the experiment,the overall tendency was that two groups of samples were fist clustered in its own group,and then combined with the other,but the second part of the experiment was more obvious.4.The results of two experiment all suggest that the relative abundance of Firmicutes was decreased and Proteobacteria was increased,But the second experiment was affected more deeply,LefSE analyse was used to compared the intestinal flora difference between before and after surgery,In the first experiment,pl.bacteria were found statistical difference in class?order?family?genus level,The vast majority of the decline belongs to Lachnospiraceae,and Ruminococcus,only bacteria increased was Atopobium.In the second part of the experiment,in addition to classe,order,family,genera level had statistical differences,Proteobacteria increased significantly,further refinement,in addition to the further lost relative abundance of lachnospiraceae,and Ruminococcus,the other phylums was also affected,and after the operation,Escherichia,Enterococcus become dominant bacteria.Conclusion:1.16 S rDNA sequence analysis technique could detect ahnost all the flora in the feces,and enables us to have a more comprehensive understanding of the composition of kidney transplantation patients intestinal flora.2.the process of renal transplantation can damage the diversity of intestinal flora,and the continued use of sulfa drugs can lead to significant loss of diversity3.the commensal bacteria was decreased,and opportunistic bacteria was increased,obviously in patients which take sulfa drugs.This flora disorder can induced metabolites disorder,bacterial translocation,immunologic derangement,and further lead to infection or even rejection on the basis of intestinal mucosa damage,But this conclusion needs further experimental verificat.