Based On The Macro Genomics Gallbladder In Patients With Gallbladder Stone Molecular Information Of The Microbial Community Analysis

Objective:To explore the role of microbial communities in the gallbladder stone formation mechanism.explain this study has based on clinical research, according to the center of the change of the surgical treatment of gallbladder calculi in nearly a decade, using new technology, a comprehensive system of calculi in patients with gallbladder gallstone disease microenvironment and the mucous membrane of the microbial community structure and composition analysis, For the further study of microbial community in the gallbladder stone formation research provides important theoretical basis and foundation of work, provide certain reference for preventing clinically to gallstones.Methods:Chapter 1 Retrospective analysis of 2004-2014 period underwent surgical treatment of patients with gallbladder stone and analyze the clinical data Chapter 2 Depending on the amount of cholesterol in the calculus is different, respectively collected 20 cases,20 patients with cholesterol stone bile pigment and 20 cases of patients with mixed stones stones and mucous membrane samples,11 patients with anterior right liver hemangioma and collected bile specimens as normal controls; Extract all samples of microbial community, DNA amplification by polymerase chain polymerization, using the second generation of sequencing technology Illumina Miseq) on a large scale sequencing of the microbial community, identified microbes OUT. According to the identification of microbial species, heat maps, network analysis and functional annotation of similar species genome means, analysis and comparison of the different components of cholesterol calculus and mucous membrane of bacteria, the community structure.Result:Chapter 1 ncluded in this study, a total of 46909 patients with gallbladder stone surgery patients with gallbladder stone increases year by year women were slightly more than men, but the composition than there was no significant difference between the two; All ages in patients with 41 to 50 years old, most patients with reduced year by year, in patients undergoing open cholecystectomy in patients undergoing laparoscopic surgery and increased year by year, common complications mainly include:cholecystectomy bile leakage and bile duct injury, abdominal abscess, lung infection. Chapter 21 control sample to detect the DNA of the microbial community, stone sample to detect the bacterial DNA in patients with gallbladder stone, detection rate of 100%, mucosal samples to detect the bacterial DNA, detection rate is 100%; Article 2 received 2177063 high quality bacterial 16 s rRNA gene sequences, each sample 22444+11729, covering 97 samples. Including 294477 cholesterol stone series, cholesterol mucosa obtained 506514 sequence; Bile pigment calculus 264534 sequence, bile pigment mucosa obtained 538507 sequence; Article 319907 sequence the mixed stone, mixed mucous membrane to obtain 253124 sequence. OTU identified 96828 bacteria, including more than two samples Shared OTU number is 18843. The average sample coverage was 86.6+/-3.8% of species. In regardless of the type of sample, the most dominant the rest of the door of the gate of Proteobacteria deformation bacteria abundance most 3 door gate of Firmicutes thick wall of bacteria, Bacteroidetes Bacteroidetes and Actinobacteria actinomycetes. To account for more than 1% of other rare bacterial categories as you can see, the same type of patients with stones and mucous membrane composition is relatively consistent, and different types of differences.3 the bacteria distribution of cholesterol stone is relatively concentrated, differences in most large groups have higher distribution, mainly concentrated in Pseudomonas bacteria which bile pigment calculus; Stutzeri and Enterococcus groups.Conclusion:1. This study adopted the cultivation for the first time the second generation sequencing technology Illumina Miseq) on a large scale of gallbladder sequencing of the microbial community, through the analysis of massive gallbladder bacterial 16 s rRNA genes, and for the first time from the system level more fully reveal the gallbladder stone formation related the basic characteristics of microbial communities.2. This study first to reveal the different cholesterol levels between patients with gallbladder stone calculus and mucosal microbial community distribution and diversity.3. The gallbladder stone formation may not be a single species but multiple comprehensive multi-factor effect between flora.