| Objective:The purpose of this paper is to study the Helicobacter pylori(Helicobacter pylori,H.Pylori)after infection,patients with upper gastrointestinal symptoms of chronic gastritis of gastric mucosa flora structure,diversity and so on will change,and analyze the H.Pylori gastric mucosa flora changes after infection with chronic infection can’t remove,gastric mucosal chronic inflammation activity of inflammatory mechanisms such as whether there is a potential link.It is of great significance to study the pathogenic mechanism and prevention of H.pylori infection.Methods:A total of 26 patients with chronic gastritis with upper gastrointestinal symptoms from June 2020 to November 2020 were included in this study,including patients with H.pylori positive chronic gastritis and patients with H.pylori negative chronic gastritis.Through questionnaire survey and oral research content,to conform to the standard set and signed agreement of patients by isotope labeling of 13 c Urea Breath Test(Urea Breath Test,UBT),Rapid Urease Test(Rapid Urease Test,RUT),gastric mucosal histopathological staining detection,if UBT positive and(or)in patients with positive RUT and gastric mucosal histopathological staining visible H.Pylori(that is,three criteria in accord with including gastric mucosal histopathological staining positive two standard),The patient was determined to have H.Pylori infection(H.Pylori positive),According to H.Pylori test results 26 patients were divided into H.Pylori positive chronic gastritis group and H.Pylori negative chronic gastritis group,and collect the samples of gastric mucosa,the use of high-throughput sequencing technology based on Illumina bacteria to 16 srrna gene(16 srrna gene,16 srrna)sequencing method,for H.Pylori infection and different inflammatory degree of patients with chronic gastritis stomach flora analysis structure,species diversity,etc.Results:A total of 26 samples were collected from patients with H.pylori positive chronic gastritis and patients with H.pylori negative chronic gastritis.After high-throughput sequencing of 16 S r RNA＿HP＿The average number of effective sequences in the POS group was 63830,and the average number of effective sequences in the POS group was 63830＿HP＿The average number of effective sequences in neg group is 68151,and the final number of operational taxonomic units(OTUs)is 11731.According to OTU analysis,group＿HP＿The amount of OTU in neg group was higher than that in group＿HP＿In the POS group,the total number of OTUs in the two groups was 5706,which accounted for 50% of the total number of OTUs in the group＿HP＿Neg group and group＿HP＿63.22% and 67.83% of the total OTUs in the POS group,which indicated that the two groups had the same flora structure to a large extent,and the high number of OTUs also suggested that the gastric flora of the two groups had high richness.Two groups of samples α The diversity index evaluation confirmed that most of the bacteria in the stomach were covered by the sequencing volume and depth of the samples,and the species richness and evenness were good α After the analysis of diversity index,we found that＿HP＿POS group and group＿HP＿The difference of gastric mucosal flora between neg group and control group was statistically significant α There was no significant difference in diversity.By further analyzing the beta diversity of gastric mucosal flora structure,we found that group＿HP＿POS group and group＿HP＿The microbial community structure of NEG group was similar,and the gastric mucosa flora of NEG group and neg group were similar β There was no significant difference in diversity.We annotated the OTU to each taxonomic level,and the gastric microorganisms were divided into 32 phyla,84 classes,207 orders,399 families,973 genera and 454 strains.Group＿HP＿POS group and group＿HP＿The dominant phyla in neg group were Bacteroides,proteus,Firmicutes,actinomycetes and epsilon bacterium aeota.The top five genera of the two groups were lachnospiraceae NK4A136＿Group),Helicobacter,Bacteroides,Klebsiella and Pseudomonas.Among them,Helicobacter is in group＿HP＿The abundance of POS group was significantly higher than that of group＿HP＿In the neg group,Pseudomonas was the most important bacteria.Then Wilcoxon difference statistical analysis was used to get the conclusion＿HP＿POS group and group＿HP＿There were significant differences in the level of phylum,class,order,family,genus and species in neg group.The differential phylum were epsilon bacterium eota and Fusobacterium;The top ten genera with different species richness were Helicobacter and christensenellaceae＿R-7＿Group,Desulfovibrio,cetobacterium,Prevotella,Brevibacillus,promonospora,pseudoxanthomonas,dialist and Micromonospora.We use genus level correlation heat map to analyze the correlation between different genera,and find that there is a certain positive and negative correlation between the genera,among which the following genera are positively correlated: Helicobacter and rikenella CEAE＿RC9＿Gut group,Bacteroides;Lachnospiraceae＿NK4A136＿Group is closely related to the genus rhodozyme,odoribacter and Prevotella＿1.Sphingomonas,alistipes,Ruminococcus 1;Pseudomonas,Enterobacteriaceae,Klebsiella;Bacteroides and rikenellaceae＿RC9＿gut group;Klebsiella is closely related to Enterobacter,parasutterella and Citrobacter;There was a negative correlation between Helicobacter and luminococcaceae＿UCG-014;Pseudomonas is closely related to faecalis;Klebsiella and alistipes,Clostridium rumen 9,Prevotella.Finally,we use picrust software to predict the function of KEGG based on 16S＿There was no significant difference in L1 level between the two groups;In KEGG＿In the metabolic pathway at L2 level,group＿HP＿POS group and group＿HP＿There was significant difference in circulatory system between neg group and control group＿HP＿In the POS group,cancer,neurodegenerative diseases,immune system,signal molecule interaction,replication and repair,circulatory system,cell movement,cardiovascular disease,cell growth and apoptosis,translation,endocrine system,environmental adaptive metabolic pathway showed an increasing trend.In KEGG＿Group at L3 level＿HP＿POS group and group＿HP＿Neg group had significant differences in seven aspects: calcium reabsorption regulated by endocrine and other factors,salivary secretion,gastric acid secretion,sodium reabsorption regulated by aldosterone,amoeba disease,Parkinson’s disease and myocardial contraction.In the process of comparing Ko values between groups,we found that group＿HP＿The POS group was better than the group in the following metabolic functions＿HP＿Neg group samples had higher expression: they were used to describe bad characteristics and predict general function;Amino acid metabolism,exogenous biodegradation and metabolism,PAHs degradation;Environmental information processing,membrane transportation,means of transportation.Conclusion:After H.pylori infection in patients with chronic gastritis,gastric flora disorder will occur,some pathogenic oral flora and intestinal flora will increase,and there are also some positive and negative correlation with H.pylori flora,which provides a new research direction for the next use of probiotics in the treatment of H.pylori infection. |
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