Correlation Between Natural Prognosis Of High-risk HPV Infection And Vaginal Microecology

Cervical cancer is one of the most serious diseases that threaten the health of women all over the world.The persistent infection of high risk human papillomavirus(HR-HPV)has been identified as a risk factor for cervical precancerous lesions and cervical cancer.After continuous infection of HR-HPV,healthy women may lead to dysfunctional vaginal microflora and cause vaginal microbiological rings.The disorder of the border,if the body has not been cleared by itself,may have progressed to precancerous or cervical cancer for several years,so the study of HR-HPV infection and vaginal microecology has attracted much attention.First,the vagina microecology is a complex system,which is regulated by the structure of the bacteria group,the local immunity and the endocrine of the body.In the healthy female vagina,more than 50 kinds of microorganisms are parasitized,mainly lactobacillus,mutual interaction and mutual restriction to maintain the balance of the vagina microecology.When HPV is infected,the number of dominant bacteria of Lactobacillus vagus decreases,while the increase of other pathogenic bacteria may lead to the imbalance of vaginal flora.Secondly,when the balance of the vaginal microenvironment is broken,the infection of the genital tract may be aggravated.Therefore,the clear relationship between the HPV and the common pathogens in the genital tract and whether the natural removal of HPV are affected by these pathogens can provide a powerful basis for the effective prevention of cross infection of various reproductive tract pathogens in women.Meanwhile,it provides a theoretical basis for further reducing the incidence of cervical cancer caused by persistent infection of HPV.Finally,the inflammatory cytokines(INF-gamma,interleukin)in vaginal microcirculation play a crucial role in maintaining the balance of vaginal microecosystem.Therefore,the further study of the correlation between inflammatory cytokines and the infection of HPV and vaginal pathogens is of great significance in preventing the infection of various kinds of pathogens in female genital tract and the occurrence of cervical lesions.Part Ⅰ:Correlation between high-risk HPV infection and vaginal bacterial dysstructureObjective:1)The structure of vaginal flora was identified by 16S rDNA sequencing,and the correlation between HR-HPV infection and vaginal microecological imbalance was discussed.Method:1)HR-HPV test and Amsel standard method were performed on subjects from outpatient department of genecology,then the subjects were divided into four groups:① 106 women with HR-HPV positive(HR-HPV+)and bacterial vaginosis negative(BV-);② 82 women with HR-HPV+ and bacterial vaginosis positive(BV+);③ 47 women with HR-HPV-and BV+;④ 59 women with HPV-and BV-.2)DNA was extracted by Bacterial DNA kit.3)16S rDNA was amplified and sequencing by PCR amplification and Illumina Miseq sequencing.4)the sequence data were uploaded to NCBI database to identify and annotate microbe,thenanalyze Principal Component Analysis(PCA).5)the microflora structures were analyzed to investigate the difference among four groups.Results:1)Vaginal lactobacillus,with ratio of 92.5%,is a kind of major and predominant microbe in healthy women,other microbes also existedin vaginal with small majority;comparing with HPV-BV+ patients,the decrease of lactobacillus,the increase of Gardnerella,Atopobium and Prevorella could be obviously seen in HPV+BV+ patients,moreover,HR-HPV+ BV+patients had higher richness of Gardnerella,Atopobium and Prevorella than HR-HPV+ BV-patients.2)PCA showed Lactobacillus,Gardnerella and Prevorella are three major microbes among groups.3)Women with Mycoplasma or chlamydia,Lactobacillus curl and Lactobacillus inert,are dominant in the vaginal flora.Conclusion:1)the vaginal microflora structure of healthy women is simple,mainly Lactobacillus.2)the HR-HPV infection can lead to the decrease of lactobacillus richness,and the increase of Gardnerella,Atopobium and Prevorella richness.Part II:Relationship between outcome of HR-HPV infection and genital tract pathogen infectionObjective:Objective to explore the relationship between HPV infection and other common pathogens of genital tract,and analyze the effect of mycoplasma infection on HPV outcome.Method:1)ccording to the test results of HPV,the patients were divided into 188 HPV positive group and 106 HPV negative group.The two groups were detected by Chlamydia trachomatis,Ureaplasma urealyticum,bacterial vaginosis,Candida and Trichomonas vaginalis.The results were statistically analyzed,and there was no correlation between HPV infection of reproductive tract and other pathogens infection.2)(1)HR-HPV positive and BV negative 106 people;(2)HR-HPV positive and 82 BV positive;(3)HR-HPV negative and 47 BV positive;59 four groups of HR-HPV and BV double negative groups were detected by mycoplasma detection to determine the difference of positive rate of infection in different groups.3)106 people with HR-HPV positive and BV negative were followed up for 12 months.During the follow-up period,no treatment(including oral administration and local use of vagina)was taken,but the patients were asked to pay attention to the vagina hygiene and review the HR-HPV to determine the effect of mycoplasma infection and other factors on the clearance of HR-HPV.Results:1)The HPV positive group and the HPV negative group were compared with the other pathogens in the reproductive tract.The infection of Ureaplasma urealyticum and HPV infection was significant(X2 =16.837,P<0.05).The infection of other pathogens and HPV infection did not have significant significance.2)In the selected group,the positive rate of Mycoplasma in HR-HPV positive and BV negative patients was 35.7%(45/106),and the positive rate of HR-HPV positive and BV positive patients was 54.6%(44/82).The positive rate of HR-HPV negative and BV positive patients was 28.8%(13/47),and the positive rate of the HR-HPV and BV double negative group was 13%(7/59).3)after follow-up of 106 patients with HR-HPV positive and BV negative,the natural clearance rate of HR-HPV in patients with mycoplasma infection was 47%(21/45),while the natural clearance rate of HR-HPV in patients without mycoplasma infection was 65.7%(40/61).Conclusion:1)there is a correlation between the infection of Ureaplasma urealyticum and HPV infection.The infection of Ureaplasma urealyticum can increase HPV infection.2)mycoplasma infection is the main factor hindering the natural clearance of HR-HPV.The number of sexual partners,the cleanliness of sexual relationship,and the presence or absence of reproductive tract inflammation are synergistic factors.Part III:The expression of high risk HPV infection and the expression of vaginitisObjective:Objective to explore the correlation between Ureaplasma urealyticum infection and inflammatory factors in vaginal microenvironment in high-risk HPV patients.Methods:According to the second part of the group,then the vaginal secretion from women in each group were collected to quantify the level of IL-2,IL-5,IL-10 and INF-γ by ELISA method and cervical tissues to investigate the the expression of above factors by western blot and immunohistochemical.Results:1)comparison between groups:the concentration of IL-2 in four groups was(1.01±0.25)pg/ml,(1.33±0.21)pg/ml,(1.42±0.32)pg/ml and(1.83±0.36)pg/ml,respectively;the concentration of IL-5 was(20.43±3.78)pg/ml,(22.08±8.32)pg/ml,(29.32±5.23)pg/ml and(31.18±4.76)pg/ml.respectively;the concentration of IL-10 was(2.14±0.16)pg/ml,(2.23±0.11)pg/ml,(1.84±0.08)pg/ml and(1.90±0.16)pg/ml,respectively;the concentration of INF-y was(13.56±2.23)pg/ml,(21.56±3.47)pg/ml,(32.90±3.44)pg/ml and(35.21±4.79)pg/ml.respectively.2)comparison in group:the relationship between IL-10 and mycoplasma infection has no statistic difference.However.The level of IL-2,IL-5 and INF-y correlate with mycoplasma infection.3)the expression of IL-2,IL-5,IL-10 and INF-y by imnunohistochemical and western blot are consistent with ELISA.Conclusion:1)HPV infection group,the expression level of IL-2,IL-5 and INF-in vaginal secretions decreased;2)mycoplasma infection group,IL-2,IL-5 and INF-gamma expression level in vaginal secretions decreased;3)HPV combined with mycoplasma infection group,the expression level of IL-2,IL-5 and INF-gamma in vaginal secretions decreased;4)the expression of IL-2,IL-5 and INF-gamma in cervical tissues was consistent with vaginal secretions.