Clinical Research On Duodenal Flora Of Functional Dyspepsia Patients With Spleen Deficiency And Qi Stagnation Syndrome

ObjectiveObjective to investigate the difference of duodenal mucosa flora between patients with functional dyspepsia of spleen deficiency and qi stagnation syndrome and normal people through clinical observation study,collect the patient’s history and clinical symptom information,study the correlation between duodenal flora and flora changes and clinical symptoms,mainly use the second generation sequencing technology to detect duodenal flora,superimpose faprotax advanced analysis,analyze duodenal flora Function.MethodsFrom 2020 to 2021,20 cases of functional dyspepsia with spleen deficiency and qi stagnation syndrome and 5 healthy subjects were collected from the people who underwent gastroscopy in the endoscopic center of Ershadao branch of Guangdong Provincial Hospital of traditional Chinese medicine.The duodenal mucosa was collected under gastroscope and the contents of the descending duodenum were obtained by cell brush.The 16S rDNA of the mucosal flora of patients and healthy subjects was sequenced by the second generation high-throughput sequencing technology.Lefse,faprotax and SPSS were used to analyze and compare the differences of duodenal flora.Results1.Basic information researchThere were no significant differences in gender,age,course of disease and BMI between the two groups(P>0.05).2.Study on duodenal flora2.1 study on duodenal flora in patients with functional dyspepsia and healthy peopleA total of 2495968 sequences were detected in 25 samples,with an average of 99838 sequences per sample.The average Q20 value was 98.20%,and the average q30 value was 94.34%.On this basis,we compared the a diversity of intestinal flora between patients with functional dyspepsia and healthy people.The results showed that there were significant differences in ace index,Shannon index and observed species index of intestinal flora between patients with functional dyspepsia and healthy people(ACE,P=0.00026;Shannon,P=0.02364;observed species,P=0.0008).Then we used PCoA to analyze the structure of the two groups of flora,and found that the duodenal flora of the two groups of patients with functional dyspepsia and healthy people showed a trend of separation.The healthy samples were mainly concentrated near the origin,while the samples of patients with functional dyspepsia were relatively scattered,suggesting that there were some differences between patients with functional dyspepsia and healthy people.AMOVA analysis showed that there was significant difference between the two groups(P=0.021).Next,we analyzed the composition of duodenal flora in patients with functional dyspepsia and healthy people.At the phylum level,Firmicutes,Bacteroidetes and Proteobacteria accounted for 61.93%and 49.90%of the total sequences in FD patients and healthy people,respectively,and were the dominant bacteria of duodenal flora.At the genus level,lefse technology was used to compare the different bacterial groups,including alloprevotella,Peptostreptococcus,sutterella and lachnospiraceae_NK4A136_There were significant differences between the two groups(P<0.05).Finally,we predicted the function of duodenal flora based on faprotax software,and found that the function of duodenal flora in FD patients was significantly different from that in healthy people(P<0.05),manifested as urea decomposition and fumarate respiration,respectively.2.2 duodenal flora in patients with functional dyspepsia of spleen deficiency and qi stagnation syndromeFirstly,we compared the a diversity of duodenal mucosa flora between mild Qi deficiency patients and moderate to severe Qi deficiency patients with functional dyspepsia of spleen deficiency and qi stagnation syndrome.The results showed that there was significant difference in Shannon index(P=0.004329).Then we used PCoA to analyze the structure of the two groups of flora,and found that the duodenal flora of mild Qi deficiency patients and moderate and severe Qi deficiency patients showed a trend of separation.Further anosim analysis showed that there was significant difference between the two groups(P=0.034).The composition of duodenal flora was compared between the two groups.At the phylum level,the mild Qi deficiency group had a higher proportion of bacteroidota and spirochaetota;at the genus level,the mild Qi deficiency patients with functional dyspepsia had Prevotella,Fusobacterium,alloprevotella,Porphyromonas and Bacteroides)The results showed that the main pathogens were Treponema,faecalibacterium,sutterella and Haemophilus Dialister,Herbaspirillum and hypomicrobium were higher than those in patients with moderate to severe Qi deficiency of functional dyspepsia;the proportion of blautia and Clostridium sensu stricto in patients with moderate to severe Qi deficiency of functional dyspepsia was higher than that in patients with moderate to severe Qi deficiency of functional dyspepsia 13)The proportion of patients with mild Qi deficiency of functional dyspepsia was higher than that of patients with mild Qi deficiency of functional dyspepsia,and the results were statistically significant(P<0.05).Finally,we used faprotax software to predict the function of duodenal flora in the two groups.The results showed that patients with mild Qi deficiency had higher correlation with xylanolysis and symbiosis than patients with severe Qi deficiency.Conclusion1.There were significant differences in duodenal mucosa flora between patients with functional dyspepsia and healthy group,including large diversity of flora,different structure of flora,different composition of flora,specific taxa and specific function of flora,which predicted urea decomposition and fumaric acid respiration.2.There are significant differences in species diversity,flora structure,flora composition and specific flora function between mild Qi deficiency patients and moderate to severe Qi deficiency patients with functional dyspepsia.It is speculated that faecalis may be related to Yang Qi of traditional Chinese medicine.3.The disorder of duodenal microecology may be the formation mechanism of functional dyspepsia,which may be related to anaerobic respiration of anaerobic bacteria in duodenum to produce acid metabolites.