Study On The Correlation Between HPV Persistent Infection And Vmb After LEEP In Postmenopausal Women With High Intraepithelial Cervical Disease

Objective:In this study,16 SrRNA high-throughput sequencing was used to analyze the composition and diversity of vaginal microbial（VMB）in postmenopausal women with high cervical intraepithelial disease（HSIL）infected by HR-HPV after loop electrosurgical excision procedure（LEEP）between HPV-positive patients and HPV-negative patients.To explore the relationship between HPV infection and VMB after LEEP in postmenopausal women,to manage postmenopausal women after LEEP from the perspective of vaginal flora,and to provide effective theoretical basis for accelerating the HPV transformation and effectively preventing HSIL recurrence.Methods:1.Subjects’ enrollment and vaginal secretion collection: Strictly according to the inclusion and exclusion criteria,15 menopausal patients with pathological diagnosis of HSIL and persistent HPV infection after LEEP were included in the Case group,and 15 menopausal patients with pathological diagnosis of HSIL and negative HPV after LEEP were included in the B group.Vaginal swabs were used to collect posterior fornix secretions from the subjects,and the collected vaginal secretions were sealed and stored in a-80℃ freezer.2.Clinical data collection of subjects: HPV results before and at 3,6 and 12 months after surgery（in two follow-up visits,all patients showed positive HPV test,and the same type of infection as before surgery was regarded as HPV persistent infection,and all patients with negative HPV test were regarded as HPV negative after surgery）.3.High-throughput DNA sequencing of vaginal microbial samples: Genomic DNA of vaginal microflora was extracted,V3 and V4 hypervariable regions of16 Sr DNA were amplified,and sequenced by Illumina platform.According to species identification results,vaginal microflora was classified according to the classification method of "phylum,Class,order,family,genus and species".Bioinformatics analysis of Alpha diversity,Beta diversity,bacterial community composition and LEf Se was further completed.Results:1.Venn analysis showed that OTU number in Case group was more than that in HPV negative group,indicating that Case group had more species than B group.2.The Chao1,Shannon,Simpson and PD Whole Tree indexes of the Case group were significantly higher than those of the B group,indicating that the species diversity and richness of vaginal bacteria in the group with persistent HPV infection after LEEP were higher.3.Beta analysis showed that there were differences in vaginal microflora between Case group and B group.4.At the phylum level,the proportion of Firmicutes in group B was the highest,and that of Bacteroidetes in Case group was the highest.5.At the genus level,bacteroidetes in Case group accounted for more than lactobacillus in B group.6.At the species level,the flora of group B was dominated by Lactobacillus indolata,and the flora structure belonged to CST Type III.In Case group,anaerobic bacteria（Gardnerella,Prevotella and Atopobia vaginalis）accounted for a relatively high proportion,while lactobacillus accounted for a lower proportion,and the flora structure of this group was close to CST Type IV.Conclusion:1.The structure of vaginal microflora in postmenopausal women with high intraepithelial cervical lesions after LEEP was relatively complex in the group with persistent HPV infection,and simple in the group with negative HPV,suggesting that the persistent HPV infection after LEEP was related to the diversity of vaginal microflora.2.Lactobacillus inertiosa was dominant in the HPV negative group after LEEP,suggesting that inert lactobacillus can accelerate the removal of HPV after LEEP.3.The proportion of Gardnerella,Prevotella and Atopobia in the group with persistent HPV infection after LEEP in postmenopausal women with high intraepithelial cervical lesions increased,suggesting that it was related to persistent HPV infection after LEEP.