| Gallbladder cholesterol stones is a common surgical digestive tract disease.For the theory of the cause of gallbladder cholesterol stones,most scholars believe that cholesterol is supersaturated in cholesterol and cholesterol crystals are precipitated.In order to further explore its etiology,a large number of studies have been carried out around the process factors regulating the secretion of cholesterol,phospholipids and bile acids from the liver.With the continuous improvement of research methods,it is found that the intestinal flora participates in the metabolism of human cholesterol and participates in the formation process of the core of cholesterol stones.Therefore,the intestinal flora may play a certain role in the formation of gallstones.Objective: The intestinal flora maintains a state of dynamic balance and plays an important role in maintaining normal human physiological activities.Changes in the number,proportion,location,and biological characteristics of the gut bacteria may cause disturbances in the intestinal flora.The dynamic balance of the broken intestinal flora will affect the health of the human body and cause adverse consequences.Illumina MiSeq is a second-generation high-throughput sequencing technology that enables rapid and accurate analysis of the abundance,diversity,and composition of the gut flora.In order to determine whether there is intestinal flora imbalance in patients with gallbladder cholesterol stones in the intestinal flora,this experiment uses Illumina highthroughput sequencing to analyze the intestinal flora of patients with gallstones and healthy people.Whether there is a difference in intestinal flora,some materials are provided for the study of the role of intestinal flora in the etiology of gallstones.Methods: After review by the ethics committee,the informed consent form was signed with the specimen collection object,and the feces of 15 gallstone patients and 15 healthy people diagnosed in Qingdao Municipal Hospital diagnosed as eligible for inclusion were collected from 2017-05 to 2018-08.The stool samples were stored in a-80 ° C refrigerator.The fecal microbial DNA extraction was performed uniformly,and then the V3+V4 region of the 16 SrRNA was amplified,and the high-throughput sequencing was performed using the Illumina MiSeq platform.The sequencing results were processed by Usearch(v.11)software,and the OTU sequence abundance statistics table was obtained to analyze the abundance(alpha diversity)and abundance of the intestinal flora.Results: 1.Through the DNA extraction and detection of 15 fecal samples from patients with gallstones and healthy people,30 specimens were all in line with high-throughput sequencing standards.A total of 30 samples were successfully subjected to highthroughput sequencing.2.A total of 69,032 effective sequences were obtained from the gallstone group,and a total of 60,543 effective sequences were obtained from the healthy person group,with a total of 129,575 effective sequences.The average OUT(operational taxonomic units)(164 ± 24)in the normal group and the average OUT(158 ± 19)in the disease group.3.Statistical analysis was performed on the abundance index chao1 and the diversity index Shannon of the two groups.The results were P>0.05,and there was no statistical difference.4.At the phylum level,14 bacterial gates were detected in both groups.Among them,Firmicutes,Bacteroidetes,Actinobacteria,Fusobacteria,Proteobacteria,and Tenericumes account for more than 99%.5.Bacteroides is a dominant genus in the gallstone group and healthy people.The relative abundance of Ruminococcaceae IS in the gallstone group was higher than that in the healthy group(P<0.01).The relative abundance of Lachnospiracea IS in the healthy group was higher than that in the gallstone group(P<0.01);Roseburia(Roche)The relative abundance in the healthy group was higher than that in the gallstone group(P<0.01);the relative abundance of Sporococcus in the healthy group was higher than that in the gallstone group(P<0.01).Conclusion: 1.The second generation high-throughput sequencing technology Illumina successfully completed the structure analysis of intestinal flora,revealing the structural changes of intestinal flora;2.There was no significant change in the level of intestinal flora in patients with gallstones.The dominant bacteria phylum was thick-walled.3.In the intestinal flora of patients with gallstones,there is a dysbacteriosis at the genus level.Roseburia,Lachnospiracea I.S.,and Sporococcus are reduced in the intestinal flora of patients with gallstones,and Ruminococcaceae I.S.is increased in the intestinal tract of patients with gallstones. |
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