| Background and Aim:Choledocholithiasis is a common and frequently occurring disease of digestive system.The incidence of choledocholithiasis in China is high and has a growing trend,which seriously endangers human health.As for the mechanism of stone formation of cholangiolithiasis,although several hypotheses have been proposed,such as bile obstruction and stasis,bacterial infection of the biliary tract,and imbalance of bile composition ratio,the specific pathogenesis has not been completely clarified,and still needs further exploration.Bile duct stones can be divided into three types according to their composition:bile pigment type,cholesterol type and mixed type.The formation mechanism of bile duct stones is different,resulting in different stone composition.The previous clinical study of our team found that Periampullary diverticulum（PAD）increased the incidence of bile duct stones and was a risk factor for the recurrence of common bile duct stones.Further analysis of the composition of bile duct stones showed that the stones in patients with enterobiliary reflux were mainly loose bile pigmented stones.It is suggested that bile duct stones are associated with enterobiliary reflux.Previous studies have reported thatβ-glucuronidase（β-G）secreted by bacteria is correlated with the formation of bile pigment stones.However,the mechanism of cholangiolithiasis formation is complex,and the formation causes of different components of cholangiolithiasis are still not completely clear,which needs further research.The objective of this study is to investigate the relationship,which between duodenal parapillary diverticulum and choledocholithiasis formation and the effect of duodenal diverticulum on the composition and diversity of biliary tract flora.Methods:16 patients underwent ERCP for the removal of cholangiolithiasis at our hospital between Jan 2020 and Oct 2021 were included in this study,including 8patients with PAD and 8 patients without PAD.The cholesterol and bilirubin components in stone samples were quantitatively analyzed by spectrophotometry.Bile was collected from the two groups,and DNA of bacterial community samples was extracted.V3-V4 variable regions were amplified and library was constructed.Sequencing was performed based on Illumina Miseq sequencing platform,and differences of sample species were analyzed at different classification levels.Finally,the correlation between different bacterial flora and bile pigment and cholesterol was analyzed.Results:The difference in age was not statistically significant,as well as bile duct diameter,stone diameter,TBIL,AST,ALT and ALP between PAD group and non-PAD group（p>0.05）.After comparative analysis of cholesterol and bile pigment ratio between PAD group and non-PAD group,it was found that cholesterol content of bile duct stones in PAD group was significantly lower than that in non-PAD group（p=0.0061）,and bile pigment content in bile duct stones in PAD group was higher than that in non-PAD group（p=0.0472）,with statistically significant differences.There was no statistical significance inαdiversity index between PAD group and control group（p>0.05）.There were differences in species community composition between the two groups of samples,some of which were in the state of aggregation,and there were certain similarities between the samples.Rothia,Streptococcus,Porphyromonas,Bilophila,Clostridium,Bdellovibrio,Micrococc The abundance of aceae and Streptococcaceae in PAD group was higher than that in control group,while the abundance of Eubacterium and Lactobacillus in control group was higher than that in PAD group.The differences in 5 pathways between PAD group and control group were statistically significant.（p<0.05）,They are Biodegradation and biosynthesis pathways of secondary metabolites and Calcium signaling,respectively Pathway,Methionine and cysteine metabolism,Lipid metabolism,Taur.ine and hypotaurine metabolism Metabolism.There was a correlation between choleophilia and porphyromonas with bile pigments（R~2=0.7013,p=0.0001,R~2=0.2868,p=0.0325）.Conclusion:Choledocholithiasis patients with PAD had higher levels of bilirubin and lower cholesterol.The presence of PAD changes the diversity,and composition and abundance of biliary flora,and may affect the composition of common bile duct stone through the change of flora. |
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