| Background:Gallbladder stones and gallbladder polypoid are both common and frequently-occurring diseases in hepatobiliary surgery.Gallbladder cholesterol stones and gallbladder cholesterol polyps are the main types of these diseases.At present,the pathogenesis of cholesterol gallstones and polyps is not completely clear,and whether the formation mechanisms of the two are related is also quite controversial.However,previous studies have shown that gallbladder bile supersaturation is one of the basic pathological mechanisms of its formation.In recent years,with the development of molecular biotechnology,more and more evidences show that the intestinal flora promotes the formation of various types of gallbladder stones,and whether the gut microbiota is related to the formation of gallbladder polyps,there is currently no relevant research.Purpose:This study intends to use high-throughput sequencing technology to comprehensively describe the composition and structure of the gut microbiota of healthy people,gallbladder cholesterol stones and cholesterol polyps patients;to explore the gut microbiota structure through a comparative analysis of the three groups of people whether the tract microbiota is related to the onset of cholesterol gallstones and cholesterol polyps in the gallbladder;clarify whether the gut microbiota of patients with cholesterol gallstones and cholesterol polyps have some similar characteristics or significant differences,so as to provide the theoretical basis and follow-up research supplements and lays the groundwork,and at the same time provides us with new ideas and basis for clinical prevention and treatment.Methods:A total of 35 patients who were hospitalized for cholecystectomy at General Surgery Hospital of Shaanxi Nuclear Industry 215 Hospital from February 2020 to June 2020,and were confirmed to be cholesterol stones and cholesterol polyps by pathology after the operation,of which the gallbladder cholesterol stones group 29 people,6 people in the gallbladder cholesterol polyp group.In the healthy control group,a total of 27 health examiners were selected during the same period.All research objects should meet the criteria for inclusion and exclusion.After admission,the first feces of the disease group were collected in the early morning before the operation after admission,and the first fecal samples of the healthy group were collected in the early morning and frozen in an ultra-low temperature refrigerator at-80°C within 30 minutes.Then,the samples were subjected to DNA extraction,16 S r RNA V3+V4 region amplification,and finally the Illumina Hiseq2500 sequencing platform was used for high-throughput sequencing,library construction and biological information analysis.Result:1.The detection rate of bacterial DNA in all samples collected this time is 100%.2.A total of 2819414 high-quality tags were obtained for all samples,of which the healthy group received a total of 1245171 tags,the cholesterol stone group received a total of 1246729 tags,and the cholesterol polyp group received a total of 283432 tags.Based on97% sequence similarity,a total of 695 bacterial OTUs were identified after the database was compared.Among them,589 OTUs were identified in the healthy group,621 OTUs in the cholesterol gallstone group,and 394 OTUs in the cholesterol polyp group.3.There was no significant difference in the statistical analysis of the alpha diversity index of the three groups of samples(all P>0.05);in the beta diversity analysis,there was a significant difference between the beta diversity groups based on the unweighted Unifrac distance(P=0.0009);In the PLS-DA analysis,the three groups of samples were grouped into three categories,indicating that there are differences in the flora structure among the three groups of samples.4.At the phylum level,the three groups of samples with the most dominant phyla include Firmicutes,Bacteroides,Proteobacteria,and Actinobacteria;the cholesterol stone group has the highest relative abundance of Firmicutes;the cholesterol polyp group has the highest relative abundance of Proteobacteria and the relative abundance of the Bacteroides phylum is the highest in the healthy group.5.At the genus level,the most dominant bacterial genera of the three groups of samples include Bacteroides,Prevotella,Faecalibacterium,Salmonella,and Escherichia.The relative abundance of Escherichia in the cholesterol stone group was the highest;the relative abundance of Prevotella and Salmonella in the cholesterol polyp group was the highest.6.In the LEf Se analysis,the Verrucomicrobia of the cholesterol polyp group increased significantly from the phylum level to the family level(including Akkermansia).Conclusion:For the first time in this study,the Illumina Hi Seq platform was used to simultaneously perform high-throughput sequencing of the gut microbiota of healthy people,gallbladder cholesterol stones and cholesterol polyps patients,and demonstrated the diversity and composition structure of their gut microbiota.This study found that there was no significant difference in the alpha diversity of the gut microbiota,while the beta diversity had significant differences.This study found that the main dominant microbiota of the three groups of populations were the same regardless of the phylum level or the genus level,but the relative abundance of the microbiota was different between the two groups.Compared with the healthy group,both the gallbladder cholesterol gallstone group and the cholesterol polyp group had bacterial microbiota imbalance.Gut microbiota imbalance is related to the occurrence and development of cholesterol gallstones and cholesterol polyps in the gallbladder;this study found for the first time the relative abundance of Verrucomicrobia(including Akkermansia)from the phylum level to the genus level in patients with cholesterol polyps in the gallbladder both are significantly increased,which may be used as a biomarker for the diagnosis of gallbladder cholesterol polyps. |
