Effect Of Qingdu Suppository On Vaginal Flora In Patients With Persistent Cervical HR-HPV Infectio

Purpose:In this study,16S rdna sequencing was used to analyze the differences in vaginal flora between healthy women and women with persistent high-risk human papillomavirus(HPV)infection,and the changes of vaginal microflora after treatment of high-risk human papillomavirus persistent infection with traditional Chinese medicine Qingdushuan for three months.The mechanism of Chinese medicine in improving the occurrence and development of vaginal microecology was also analyzed,it provides a new idea for the treatment of high-risk HPV persistent infection with traditional Chinese medicine.Method:1.Vaginal discharge collection and study participants:we collected healthy women aged 20-45 years who were admitted to Beijing University of Chinese Medicine Oriental Hospital between April 2022 and December 2022,the vaginal posterior fornix secretions were obtained with informed consent and signed informed consent form.The patients with persistent high-risk HPV infection were treated with Qingdushuan for three months and then vaginal discharge again.30 cases in each Group,and named the Healthy Group as the healthy control Group(Group C),and the high-risk HPV persistent infection Group as the H Group before treatment,after treatment,the patients were named He Group and the vaginal discharge were stored in a-40 C refrigerator for later use.2.Data Collection:The subjects’ pregnancy and delivery history,age,height,Body Mass Index(BMI),age at first sex,Human papillomavirus of HPV infection,Thinprep cytologic test(TCT),hematuria,electrocardiogram,liver and kidney function were recorded。3.16S rdna sequencing of vaginal discharge flora:16 subjects in Group C,27 subjects in Group H and 14 subjects in group He were included in this study.The vaginal discharge of the subjects were used for 16S rdna sequencing analysis.According to the results of sequencing,the vaginal flora was identified and classified according to the classification method of"Phylum,class,order,family,genus and species".4.Statistical analysis:SPSS Software was used to analyze the clinical data of the study subjects,GrapHpad prism 8.0 software was used to analyze the differences of vaginal microbial diversity among the study subjects,and further analysis of "Qingdu suppository"treatment before and after the change in vaginal flora diversity.Results:(1)HPV group had 5427 OTUs,C group had 3265 OTUs and He group had 1470 OTUs.(2)at the level of phyla and genera,Firmicutes and Lactobacillus were in a descending order in Group C,HPV and He,and prevotella and streptococcus were significantly enriched in HPV group Gardnerella was significantly enriched in Group C,the abundances of Streptococcus,mirabilis,Stenotrophomonas,Megasphaera,Gardnerella and Lactobacillus decreased.(3)there was no significant difference in the Shannon Index and Simpson Index of Aphla Diversity Index among the three groups(P>0.05),and the Shannon Index and Simpson Index of the HPV group were higher than those of the C group.(4)NMDS map and PCOA analysis shows,there are differences in vaginal microbial βdiversity among the three groups.(5)LLEFSE analysis(LDA>4),4 Proteobacteria,Rhizobiales,Methylobacteriaceae and Methylobacterium were screened from Group C,in the HPV group,Streptococcus,Gammaproteobacteria,Coriobacteriales,Coriobacteriaceae,coriobacteriia and Atopobium were screened,In He group,potential bacteria were identified as markers of Bacteroidales,Bacteroidetes,Bacteroidia,Prevotella,Prevotellaceae,Mollicutes and Tenericutes.(6)the analysis of interspecies relationship network showed that most of the interrelationships were among Firmicutes,bacteroidetes,Actinomycetes and Proteobacteria The antagonism was mainly concentrated among Lactococcus lactis,Weissella and Lactobacillus.(7)comparing the predicted gene function of KEGG database,20 functional metabolic pathways were predicted in all three groups at Level 3,they include Homologous fixation in photosynthetic organisms,recombination,Protein export,Cell cycle-Caulobacter,Ribosome,Aminoacyl-tRNA biosynthesis,Mismatch repair,D-Glutamine and D-glutamate metabolism,and alanine metabolism.There was no significant difference among the three groups.Conclusion:(1)there were significant differences in vaginal microbial diversity between healthy women and women with persistent high-risk HPV infection,and the highest diversity was found in women with persistent high-risk HPV infection.(2)traditional Chinese medicine QingduShuan can Adjustment vaginal microbial diversity,QingduShuan may affect the abundance of "Beneficial bacteria" such as Lactobacillus,it also reduced levels of bacteria such as gardnerella,Streptococcus,mirabilis,Stenotrophomonas,and chlorella,but was not susceptible to prevotella.Therefore,when using Qingdu suppository to treat HPV infection,it may be appropriate to cooperate with Lactobacillus preparation and adjust the flora.(3)Lactobacillus can maintain vaginal homeostasis,inhibit the invasion of pathogenic microorganisms and inhibit the persistent infection of HPV.Increased abundance of prevotella,streptococcus,mirabilis,Stenotrophomonas,and Chlorella may contribute to increased susceptibility to HPV.(4)single Gardnerella may can not cause disease,and it is possibly the joint effect of many kinds of pathogenic bacteria that cause low reproduction and infection.(5)vaginal microecology is a complicated microenvironment formed by the cooperation and antagonism of many kinds of microorganisms.It is necessary to detect many kinds of pathogenic bacteria in order to make more accurate diagnosis,reflect the more realistic composition of the vaginal flora.(6)the predominant strains of HPV group were Streptococcus,Coriobacteriales,Coriobacteriaceae,coriobacteriia and Atopobium,these bacteria may serve as potential diagnostic markers of vaginal dysbacteriosis and disease risk in patients with persistent high-risk HPV infection and may provide insights into the microecological treatment of patients with persistent HPV infection of the cervix.(7)Qingdu suppository may help to eliminate HPV infection from two aspects.Firstly,Qingdu suppository can help to restore vaginal environment and reduce HPV susceptibility through its bacteriostatic and anti-inflammatory effect;The other is to eliminate HPV infection directly through the effect of anti-virus.