A Comparative Study Of Bile Duct Flora In Patients With Common Bile Duct Stones And Cholangiocarcinoma

Objective:Through high-throughput 16 s rDNA sequencing to compare the structure and composition of the microbial community in the bile of patients with common bile duct stones(CBDS)and cholangiocarcinoma(CCA),as well as the characteristics of the core biota,to explore the role of microorganisms in the pathogenesis of common bile duct stones and tumors,While looking for more effective identification and prevention methods.Methods: 1.Collect the patients diagnosed with common bile duct stones or cholangiocarcinoma from December 2019 to December 2020 in the Affiliated Hospital of Yan’an University.Patients who meet the inclusion criteria are divided into common bile duct stones group(group C)and cholangiocarcinoma group(group D).The common bile duct stones group passed the standard endoscopic retrograde cholangiopancreatography(ERCP)aseptic operation.The bile samples were collected by the catheter,and then immediately put into the standard preservation solution(Effc Gut)for preservation;patients with cholangiocarcinoma were withdrawn from the percutaneous transhepatic cholangial drainage(PTCD)drainage tube,and then immediately put into the standard preservation solution(Effc Gut)for preservation;2.Using Illumina high-throughput sequencing method forDNA extraction of bile microorganisms and gene sequencing and bioinformatics analysis of the V4 region of 16 S rDNA;3.Analyze and compare the species with differences in bile flora between common bile duct stones and tumor patients;4.Use SPSS22.0 statistical analysis software for data analysis;measurement data conforming to normal distribution are represented by mean ± standard deviation;comparison between groups is by t test;The statistical data were tested by χ2 test.Counting data that did not conform to the normal distribution were represented by the median(P25,P75),and the rank sum test was used for comparison of two independent samples.P<0.05 was considered statistically significant.P<0.05 indicates that the difference is statistically significant.Results:This study included 25 cases of research objects,including 19 cases of common bile duct stones,6 cases of cholangiocarcinoma groups.The data of all patients are analyzed,and the results showed that: 1.There is no statistical significance in general data of the patients(P > 0.05);The differences of TBIL,DBIL,IBIL,CA199 and ferritin in laboratory data are statistically significant(P < 0.05);2.The biliary flora is rich in microbial communities and has individual heterogeneity.The two groups have high abundance at the phylum level: Firmicutes,Proteobacteria,Bacteroidetes,Actinobacteria,and Fusobacteria;3.There is no statistical significance of the abundance,diversity and uniformity of microbial community in patients with bile duct stones and bile duct cancer(P> 0.05);4.There are significant differences in flora structure and species composition between the two groups.At the phylum level,the abundance of Firmicutes,Bacteroidetes and Actinobacteria in common bile duct stones group is higher than that of cholangiocarcinoma,while the abundance of Proteobacteria and Cyanobacteria in cholangiocarcinoma is higher;5.The characteristic communities f common bile duct stones and cholangiocarcinoma are found to be different between the two groups through the analysis of species differences.The characteristic flora of the choledocholithiasis group are:Actinomycetaceae,Actinomycelales,Coriobacteriales,Coriobacterlla,Enterococcaceae,Streptococcaceae,Lactobacillales,Bacilli;The different flora in the cholangiocarcinoma group are: Sporichthyaceae,Frankiales,Burkholderiaceae,Betaproteobacteriales;6.The difference in microbial community function genes on the two groups is significantly different,and the functional gene of cholangiocarcinoma group is significantly higher than the common bile duct stones.Conclusion: 1.The biliary tract has a highly rich microbial colonies and has heterogeneity between individuals;2.The biliary microbial community is similar to the oral cavity and the duodenal microbial colonies;3.There are differences in the structure,composition,characteristic flora and functional genes between common bile duct stones and cholangiocarcinoma,suggesting that changes in biliary flora may be the pathogenic factors of different biliary diseases.