Analysis Of Vaginal Microbiota In Patients With Bacterial Vaginosis And Complication With Pelvic Inflammatory Disease

Bacterial vaginitis(BV)is a high incidence disease in women of reproductive age,which is easy to treat but easy to relapse.It can cause many female genital tract diseases and aggravate the susceptibility of sexually transmitted diseases.Pelvic inflammatory disease is a common infectious disease in female reproductive tract diseases.Research shows that female reproductive tract diseases are often accompanied by changes in vaginal microbiota.This experiment provides experimental basis for clinical diagnosis and treatment of bacterial vaginosis through detection of clinical samples,screening of pathogenic bacteria,verification of animal models,exploration of pathogenic mechanism,and probiotics intervention.Objective By comparing the vaginal microbiota of bacterial vaginosis patients and its patients with pelvic inflammation and healthy individuals,the structure and changes of vaginal microbiota were analyzed,the pathogenic bacteria were screened to establish an animal model and the pathogenic mechanism was analyzed,and probiotics were used to intervene in order to provide experimental data for clinical identification and diagnosis,laying a foundation for future related topics.Methods 1.Seventeen patients with bacterial vaginosis,13 patients with bacterial vaginosis complicated with pelvic inflammation and 52 healthy persons were screened by clinical Amsel criteria.The bacterial genomic DNA was extracted from vaginal sputum secretion by sterile swab.2.PCR technology was used to amplify the 16 S r RNA fragment obtained from the previous step.Then the amplified products were separated by deformable gradient gel electrophoresis(DGGE)to obtain the structural map of vaginal bacterial species.Cluster analysis was carried out by Quantity One software,and the specific bands were analyzed by gel sequencing.The 16 s r RNA gene was detected to specifically study the species classification in the sample.3.Special bacteria were isolated,including G.vaginallis and L.crispatus.Then,G.vaginalliswere used to establish animal models and determine the infection mechanism.Meanwhile,L.crispatus were used to intervene in animal models.4.Detection of animal model indicators,including: HE staining to detect histopathological damage;PAS staining to detect glycogen content in tissue epithelial cells;Diff staining was used to detect exfoliated epithelial cells;Realtime PCR to detect the content of immune factors,and so on.5.The experimental results of clinical samples and animal models were comprehensively analyzed.Results 1.The results of 16 S and PCR-DGGE showed that the composition of vaginal microbiota in patients with bacterial vaginosis anditswith pelvic inflammation was significantly different from that in healthy people.Among them,Firmicutes decreased,Actinobacteria increased,G.vaginallis,P.vaginallis and other anaerobic bacteria increased,L.crispatus,L.iners and other probiotics decreased.2.Animal experiments showed that vaginal infection caused by G.vaginalis produced a strong exfoliation reaction of epithelial cells,which accelerated epithelial migration,while L.crispatus could alleviate it.3.In HE staining,fallopian tube damage can be observed.It is speculated that bacteria have ascending behavior,that is,infection caused by vaginal microbiota changes can lead to ascending infection.4.Animal models showed that G.vaginalis infection could lead to changes in the expression of immune factors,and L.crispatus showed an opposite trend after the intervention of the model group.Conclusion Bacterial vaginosis itself and infection with pelvic inflammation have changed the microecological balance of vagina.Specific bacteria isolated from patients can infect mice to produce vaginitis.The pathogenic mechanism may be that pathogenic bacteria destroy the mechanical barrier of vaginal wall,and probiotics can reduce infection.