VAGINAL MICROECOLOGY IN HPV-ASSOCIATED CERVICAL DYSPLASIA

Background: The vaginal microbiota consists primarily of Lactobacillus species, which help maintain microbial balance through acid production and antimicrobial activity. The disruption of balance causes vaginal dysbiosis and infection. Objective: The aim of this study was to analyze the association of vaginal infection with the distribution of Lactobacillus species, vaginal pH, and antimicrobial activity. Methods: The study was designed as a case-control study. A total of 100 subjects, including 50 healthy subjects and 50 infected subjects, were included. Vaginal swabs were collected to assess pH, microbial isolation, and identification using the standard microbiology technique. The antimicrobial activity of Lactobacillus isolates was assessed using the agar well diffusion technique and peroxide production was assessed qualitatively. Chi-square test and independent t-test were used for the statistical analysis of the results. Results: The isolation of Lactobacillus was 90% in healthy women and 40% in infected women, with a statistically significant difference (p < 0.001). Infected women had increased vaginal pH (5.6 ± 0.5) as compared to healthy women (4.3 ± 0.3) (p < 0.001). Lactobacillus crispatus was the most frequent in healthy women and Lactobacillus iners was the most frequent in infected women. Pathogenic organisms, especially Gardnerella vaginalis, were statistically significantly more common in the infected group. The isolates taken from healthy women had the highest hydrogen peroxide production and antimicrobial activity. Conclusions: An increase in vaginal pH, a decrease in antimicrobial defenses, and overgrowth of pathogens are associated with the loss of functionally active Lactobacillus species. Composition and functional capacity are critical factors in determining vaginal health.

The association of HPV infection and vaginal microbiota of reproductive women in China: A multicenter cohort study protocol

Objective To investigate the correlation between human papillomavirus (HPV) -16, -18 and C-X-C chemokine receptor (CXCR) 7 in cervical cancer and intraepithelial neoplasm. Methods A total of 59 patients who underwent cervix resection and were diagnosed as cervical cancer by pathological detection, 86 patients who under went cervix resection and were diagnosed as cervical intraepithelial neoplasm (CIN) by pathological detection and 20 patients who underwent cervix resection due to other diseases were collected as study subjects in the Zhuhai People′s Hospital from March 2011 to March 2013. They were enrolled into cervical cancer group (n=59), CIN group (n=86) and control group (n=20), respectively. The expression of CXCR7 was examined by immunohistochemical, and the difference of CXCR7 expression among 3 groups were analyzed. Subsequently, the relative expression of HPV-16 DNA and HPV-18 DNA were verified with quantitative real-time PCR. Eventually, the protein expression profile of CXCR7 and stromal cell-derived factor (SDF)-1 were tested by Western blotting among 3 groups, and then the correlation analysis were performed between the expression HPV-16 DNA, HPV-18 DNA and CXCR7. There were no significant differences between two groups in age and other general clinical data (P>0.05). Results ①The expression positive rates of CXCR7 in cervical tissues of cervical cancer group and CIN group both were significantly higher than that of control group, which were 88.1% (52/59) vs 15.0% (3/20) and 46.5% (40/86) vs 15.0% (3/20), and both the differences were statistically significant (χ2=37.77, P<0.001; χ2=6.68, P=0.010). ②The expression of HPV-16 DNA in CIN Ⅰ, CIN Ⅱ, CIN Ⅲ and cervical cancer tissues were (1.50±0.05) times, (2.87±0.09) times, (3.17±0.12) times and (6.41±0.20) times as much as that in normal cervix tissues, respectively, and the expression of HPV-16 DNA in CIN Ⅲ and cervical cancer tissues both were significantly higher than that in normal cervix tissues, and both the differences were statistically significant (t=2.15, P=0.042; t=3.11, P=0.003). Likewise, the expression of HPV-18 DNA in CIN Ⅰ, CIN Ⅱ, CIN Ⅲ and cervical cancer tissues were (1.56±0.07) times, (2.45±0.11) times, (3.89±0.16) times and (6.12±0.17) times as much as that in normal cervix tissues, respectively, and the expression of HPV-18 DNA in CIN Ⅲ and cervical cancer tissues both were significantly higher than in normal cervix tissues, and both the differences were statistically significant (t=2.21, P=0.024; t=3.03, P=0.004). ③The expression of CXCR7 and SDF-1 increased as the the deterioration of the disease by Western blotting. Not only that, there were positive correlation between CXCR7 expression and HPV-16 DNA (r=0.74, P<0.001), and HPV-18 DNA (r=0.78, P<0.001). Conclusion HPV may contribute the progression of cervical cancer induced by high-risk HPV. Key words: Receptors, chemokine; Human papillomavirus 16; Human papillomavirus 18; Uterine cervical neoplasms; Cervical intraepithelial neoplasia

Objective To investigate the expression and clinical significance of p63, CD_(44v6) and human papilloma virus (HPV) 16/18 in carcinoma of cervix and precancerous lesion and to study their role in the pathogenesis of the infiltrative carcinoma of the cervix(ICC). Methods 145 patients were selected, among whom were 60 cases of ICC, 55 cases of cervical intraepithelial neoplasia (CIN) and 30 cases of normal cervical epidermis (NCE). Immunohistochemistry streptavidin peroxidase (SP) was used to detect the expressions of p63 and CD_(44v6) protein and hybridization in situ was used to measure HPV 16/18 gene on tissue microarray. Results HPV 16/18, p63 and CD_(44v6) in ICC were significantly higher than those in CIN and NCE groups (P < 0.05). The expressions of HPV 16/18 gene in CIN I 、CIN II and CIN III were 27.3 % , 43.8 %, 70.6 % respectively. p63 protein was mainly expressed in squamous cell carcinoma, but not in adenocarcinoma. p63-positive rate was related to the grade of squamous cell carcinoma and clinical stage. CD_(44v6) positive rate was related to the pathological grade and clinical stage. Moreover, the positive rate of lymph node metastasis was significantly higher than the cases without metastasis (P <0.05). The expression of HPV16/18 was positively correlated with that of p63 in ISCC (P <0.05, Cp =0.49). Conclusion HPV16/18 possibly participate in the pathogenesis and progress of cervical squamous carcinoma. p63, as the cancer gene, may participate in the occurrence and development of cervical cancer caused by HPV 16/18. p63 could be a differentiation indicator of cervical squamous carcinoma, and it could be one of the markers which would be differentiated into squamous cell in malignant tumour. CD_(44v6) could be used as one of the indicators of distant metastasis of cervical cancer. Key words: Uterine cervical neoplasms; Cervical intraepithelial neoplasia; Human papilloma virus (HPV); p63; CD_(44v6)

BackgroundmiR-21, miR-26b, miR-221/222 and miR-126 play crucial roles in cervical cancer development. Studies have shown that polymorphisms in miRNA genes can affect miRNA expression, which might be associated with cancer development.MethodsTen single-nucleotide polymorphisms (SNPs) in the miR-21, miR-26b, miR-221/222 and miR-126 genes (rs1292037, rs13137 in miR-21; rs2227255, rs2227258 in miR-26b; rs2858061, rs34678647, rs2858060, rs2745709 in miR-221/222; rs2297537, rs2297538 in miR-126) were selected, and genotyped in a total of 2176 individuals, including 435 patients with cervical intraepithelial neoplasia (CIN), 743 patients with cervical cancer (CC) and 998 healthy persons using TaqMan assays, and their associations with CIN and CC were evaluated.ResultsOur results showed significant differences for the rs2297538 genotypes between the CIN and CC groups (P = 0.001). In addition, our results also showed significant differences for the rs2297537 alleles between the CIN and CC groups (P = 0.003), and the C allele of rs2297537 might be associated with a decreased risk of CC (OR = 0.72, 95%CI: 0.58–0.90). At the inheritance analysis, between the CIN and control groups, the T/T-T/C genotype in rs1292037 and A/A-A/T genotype in rs13137 might be associated with an increased risk of CIN in the recessive model (OR = 1.61, 95% CI: 1.17–2.20 and OR = 1.58, 95% CI: 1.15–2.15). In addition, the C/C-T/T genotype of rs2745709 might be associated with a decreased risk of CIN in the overdominant model (OR = 0.66, 95% CI: 0.52–0.82). Between, CIN and CC group, the T/T-C/C genotype in rs1292037 and A/A-T/T genotype in rs13137 might be associated with an increased risk of CC in the overdominant model (OR = 1.43, 95% CI: 1.12–1.81 and OR = 1.42, 95% CI: 1.12–1.80). The rs2297538 G/G-A/G genotype might be associated with an increased risk of CC in the recessive model (OR = 2.83, 95% CI: 1.52–5.25). The rs2297537 2C/C + C/G genotype might be associated with a decreased risk of CC (OR = 0.71, 95% CI: 0.57–0.89) in the log-additive model. The rs2745709 T/T-C/C genotype might be associated with an increased risk of CC (OR = 1.44, 95% CI: 1.13–1.83) in the overdominant model.ConclusionOur results indicate that rs2297538 and rs2297537 in miR-126, rs1292037 and rs13137 in miR-21, and rs2745709 in miR-221/222, may have important roles in the development of CIN or CC.

Study question Can vaginal microbiota transplantation with shotgun-verified eubiotic vaginal microbiome over three menstrual cycles, convert vaginal dysbiosis to eubiosis in healthy women? Summary answer The RCT is ongoing until it is by March 2023. Results will be ready for presentation at EHSRE conference. What is known already Vaginal dysbiosis covers imbalances in the vaginal flora, caused by the composition of bacteria, viruses, and vaginal fungi. A large proportion of women who have vaginal dysbiosis do not experience any symptoms. Dysbiosis occurs in about 16-20% of all women. Vaginal dysbiosis is associated with infertility, euploid pregnancy loss, preterm labour or bacterial vaginosis. Treatment of vaginal dysbiosis consists of antibiotic treatment, and/or probiotics. Vaginal transplantation with eubiotic vaginal bacterial flora in combination with antibiotics has successfully been performed in four out of five recipients in an earlier study, but no study has been performed without use of antibiotic pretreatment. Study design, size, duration Randomized, controlled, double-blinded trial with a randomization ratio 3:1 to receive either a eubiotic microbiome transplant or placebo. 320 healthy women between 18 and 40 years, was enrolled for screening of vaginal dysbiosis. 30 donors with a eubiotic vaginal microbiome and 48 recipients with a dysbiotic microbiome were identified. The trial began in June 2021 and will end in March 2023 Participants/materials, setting, methods Vaginal microbiome composition was assed by next-generation ShotGun sequencing. To qualify as donor bacterial DNA from a vaginal swab had to show a bacterial composition of at least 80% lactobacilli and less than 5% pathogenic bacteria. To qualify as recipient vaginal microbiome composition should be with at least 20% pathogenic bacteria and no more than 10% lactobacilli. The recipient could have up to three attempts of VMT treatments with a follow-up period of 6 months. Main results and the role of chance We expect the RCT to end in March 2023, and the results of this trial will be presented at the ESHRE annual meeting . If we can show engraftment of a eubiotic microbiome transplant without the use of pre-treatment with antibiotics it could be a potential treatment of vaginal dysbiosis. Limitations, reasons for caution As this is the first VMT without antibiotic pretreatment we did not have reliable data for our power calculation and a negative result may be due to lack of statistical power. Wider implications of the findings Effective treatments for vaginal dysbiosis are urgently needed and VMT may be one such treatment strategy. Trial registration number NCT04855006

Sera from patients with cervical cancer, cervical intraepithelial neoplasia (CIN) and non-genital cancers, and from healthy individuals, were investigated for antibodies to human papilloma virus (HPV) early proteins E4, E6 and E7 and the major capsid protein LI by Western blot analysis of recombinant HPV proteins. There was a significantly higher prevalence of sera with antibodies to E6 in cervical cancer patients than in healthy individuals or in CIN or non-genital-cancer patients. Antibodies to E7 were detected in 25% of cervical-cancer patients, which is significantly higher than in HPV-associated cervical lesions or in control populations, but not significantly different from the incidence in patients with non-genital cancers. Antibodies to LI were found more frequently in CIN, while antibodies to E4 had a similar prevalence in cervical-cancer, cervical-dysplasia and non-genital-cancer groups, with 24% in the controls. The inability to detect antibodies to E6 and E7 in the majority of cervical-cancer patients limits the application of this methodology to the monitoring of HPV infection and the development of cervical cancer. However, the latter approach may be useful in combination with other assay systems which allow detection of different, including conformational, epitopes of HPV E6 and/or E7 recombinant proteins.

Objective To explore the expression of death-associated protein kinase 1 (DAPK1)in cervical carcinoma and cervical intra-epithelial neoplasia (CIN),initially investigate the relationship among DAPK1 expression and the clinical characters of the patients,then analysis the clinical significance.Methods Western blotting were used to examine the expression of the DAPK1 in the normal cervical tissue,CIN and cervical carcinoma.The relative density of the samples were examined by the high resolution pathological image analysis system (HPIAS).Results The expression of DAPK1 only can be examined in part of the CIN and cervical cancer,the relative density were 0.64 ± 0.04 and 0.36 ± 0.03 respectively and the difference between the two groups was statistically significant(P ＜0.05).The DAPK1 expressed in all the normal tissue (the relative density was 0.89 ±0.03),and it had a higher level expression than the CIN and cervical cancer tissues.The difference were statistically significant (P ＜ 0.05).The expression of the DAPK1 in the cancer tissues with no lymph node metastasis (the relative density was 0.39 ±0.03) were significantly higher than the tissues with lymph node metastasis(0.16 ± 0.04),the difference between the two groups was statistically significant (P ＜ 0.05).No relationship had been found among the expression of the DAPK1 and the age,tumor size.Conclusion The lower expression of DAPK1 may be involved in the development of cervical cancer; the DAPK1 gene may be associated with the cancer invasion and metastasis. Key words: Cervical cancer; Cervical intra-epithelial neoplasia; Death-associated protein kinase 1 ; Metastasis

Cervical cancer (CC) contributes to 6%-29% of all cancers in women. In India, 527,624 new cases of CC are added every year. India contributes to ¼ of deaths worldwide to CC it occurs often in women around the age of 30 yrs which is contributed by a specific kind of human papillomavirus causes long-term infection and inflammation (HPV) which result in morphological changes in the cells of cervix the region that connect vagina and uterus. Cervicovaginal microbiome is observed to be highly diverse among patients with CC where there is reduced number of Latobacillus spp that leads to dysbiosis and decrease in pH and eventually colonised by other anaerobic bacteria. The shift in community state types (CST) is highly associated with the Human Papillomavirus infection and its further progression to cervical dysplasia or CIN i.e. ‘Cervical intraepithelial neoplasia’ and malignant tumour of the cervix uteri. The purpose of this research is to figure out if there’s a link between the cervico-vaginal microbiota and gynaecological cancer and the review also focuses whether cervical microbiome signatures can predict the severity of infection leading to development of CC? Probiotics can be used as a potential alternative to balance the dysbiosis of the cervicovaginal environment. Hence the review summarizes the current knowledge and the interaction of different bacterial groups with Human Papilloma Virus infection and development of CC.

Background and objective: COX2, an enzyme that metabolizes arachidonic acid to prostaglandin E2 (PGE2), contributes to the development of cervical dysplasia and cancer by inhibiting the immune response in the tumor micro-environment, as well as promoting cell proliferation, angiogenesis and invasiveness. Little is known about the changes induced by the tumor and its micro-environment on circulating immune cells. We evaluated the relative baseline expression of COX2 by circulating immune cell types and whether it is different in patients with cervical dysplasia and cervical cancer compared to normal controls. Methods: Forty three patients were included: controls (n = 9), low grade dysplasia (CIN I, n = 13), high grade dysplasia (CIN II-III, n = 14), and invasive cervical cancer (n = 7) based on histopathology. Granulocytes, monocytes, and B and T lymphocytes were isolated by Ficoll-Paque density centrifugation (GE Healthcare, (Uppsala, Sweden)) followed by negative selection with MACS Miltenyi Biotec (Auburn, CA) anti-biotinylated microbeads, total RNA isolated (QIAGEN RNeasy (Germantown, MD)) and COX2 mRNA expression evaluated with reverse transcription quantitative PCR (RT-qPCR) normalized to GAPDH levels. Serum and plasma PGE2 levels were quantified by enzyme linked immunoassay (ELISA) (R&amp;D Systems (Mineappolis, MN)). Results: There were marked differences in COX2 expression between the various cell types (p &amp;lt;0.0001, Kruskal-Wallis) with granulocytes producing the highest levels, comparable to GAPDH (1.56 ± 1.46), monocytes an intermediate amount (0.083 ± 0.086), and B and T cells very small amounts (0.033 ± 0.108 and 0.017 ± 0.079 respectively). There was no significant difference in COX2 expression by disease state. B and T cells trended toward significance (p 0.055 and p 0.08 respectively) with the relative expression of COX2 increasing with disease severity. The importance of this is not clear given the very low levels of expression. Plasma PGE2 levels differed significantly by disease state (p 0.0014) with mean levels of controls: 1083 ± 722 pg/mL, low grade: 1208 ± 576 pg/mL, and high grade: 1415 ± 2127 pg/mL. Surprisingly, levels in patients with invasive cancer were low, 464 ± 182 pg/mL. Discussion: This is the first exhaustive study of baseline COX2 expression in circulating immune cell types in any HPV-associated disease to the authors' knowledge. We have previously reported that circulating monocytes from patients with HPV-induced premalignant upper airway lesions show altered immune functions in response to stimuli compared to monocytes from normal controls. Future studies are necessary to determine whether circulating immune cells from patients with cervical dysplasia and cancer show pro-tumorigenic or immune down-regulating activity in response to stimuli. Citation Format: Bethany Bustamante, James A. DeVoti, Chris J. Papayannakos, Mohd Israr, Fung Lam, Bettie Steinberg, Vincent Bonagura, Gary L. Goldberg. Circulating immune cells of patients with cervical dysplasia and cancer do not show elevated baseline expression of COX2 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr LB259.

Vaginal mucosa undergoes physiological changes across the lifespan, such as increased pH and reduced natural lubrication which are known to impact vaginal commensal microorganisms, hence increasing the chances of vaginal infections. An improved understanding of vaginal microbiome composition in different age groups and the effect of social behaviors, such as the use of personal lubricants, could facilitate the development of new strategies to maintain good vaginal health. The objective of this study was to assess the effect of water-based lubricants on the human vaginal microbiome. Fifty females from three age groups (18-29, 30-44, and 45-65 years) with mild-to-moderate vaginal dryness were randomized to one of five lubricants (four of which were formulated to meet expert guidance on osmolality and pH). Subjects entered the study at tolerance or treatment phase (vaginal intercourse minimum once a week using assigned lubricant). Four vaginal swabs per participant were sampled during pre-("baseline"), post-first ("2 h", "24 h") and post-last ("4 weeks") lubricant application to assess bacterial and fungal diversity via amplicon sequencing. Vaginal pH and relative humidity were measured at baseline, 2 h, and 24 h post-lubricant application. Relative bacteriome abundance was statistically compared between timepoints for each lubricant group. Vaginal moisture, age, BMI, and pH were correlated with bacteriome relative abundance. Lactobacilli and Gardnerella sp. Were predominant across participants. Repeated lubricant application did not significantly alter the vaginal bacteriome during 4 weeks of product use (p > 0.05) when measured by relative abundance and alpha-diversity index. Bacteriome diversity and abundance differed significantly between age groups at baseline whereas lactobacilli relative abundance was negatively associated with age and vaginal pH.

The aim of this paper was to investigate the prevalence of human papilloma virus (HPV) infection in women under 30 years old with cervical intraepithelial neoplasia (CIN) or cervical cancer, in order to provide a basis for cervical cancer prevention and treatment. We recruited 2052 female cases from the education system in Futian District, Shenzhen city, from April 2006 to April 2008, with age ranging from 22 to 60 years old. Secondgeneration hybrid capture test was done for detection of female genital tract HPV and cervical colposcopy for screening CIN or cervical cancer. The prevalence of HPV and cervical precancerous lesions or cervical cancer was evaluated in different age groups of women. The HPV positive rate was 247/2052 (12.00%), and 35/2052 (1.71%) of the cases were⩾CIN I by pathological diagnosis. In the 22–29-year-old age group, there were 291 cases, including 39 HPV-positive cases (13.40%) and one case of pathological diagnosis⩾CIN I (0.34%), and there were 1761 cases in the 30–60-year-old age group, including 208 HPV-positive cases (11.81%) and 34 cases of pathological diagnosis⩾CIN I (1.93%). The HPV-positive rate of 22–29-year-old age group was higher than that of 30–60-year-old age group, but the difference was not significant (χ 2 = 0.5967, P = 0.4398); the CIN and cervical cancer detection rate of the former group was lower than that of the latter one, and the difference was not significant either (χ 2 = 3.7519, P = 0.0527). The cases in 30–60-year-old age group were divided into five age subgroups, and the HPV-positive rate of the 22–29-year-old age group was compared with the remaining groups, and it was found that the HPV-positive rate of the former group (13.40%) was lower than that of the 40–44-year-old age group (14.70%) but higher than that of the other four groups. Moreover, the difference in the HPV-positive rate between the 22–29-year age group and the 50–60-year-old age group (6.06%) was significant (χ 2 = 5.545, P = 0.018), but the difference between the 22–29-year-old age group and each of the other four groups was not significant (P>0.05). In addition, the CIN and cervical cancer detection rate of the 22–29-year-old age group (0.34%) was lower than that of the remaining five groups, and the difference between the 22–29-year-old age group and the 35–39-year-old age group (2.26%), and between 22–29-year-old age group and 40–44-year-old age group (2.30%) was significant (χ 2 = 4.446, P = 0.0035; χ 2 = 4.525, P = 0.0363, respectively), but the difference between the 22–29-year-old age group and each of other three groups was not significant (P>0.05). Furthermore, 80.00% (28/35) cases of pathological diagnosis⩾CIN I and 90.91% (10/11) lesions of pathological diagnosis⩾CIN II occurred at 35–49 years of age. For many young women below the age of 30 who were infected with high-risk HPV were one-off infections, so it was not appropriate to recommend HPV detection as cervical precancerous lesions or cancer screening program for women less than 30 years old. The prevention and treatment of cervical cancer should be focused on those women more than 35 years of age.

Background: The vaginal microbiota, dominated by Lactobacillus species, plays a critical role in preventing infections by maintaining a low pH and inhibiting pathogens. However, antimicrobial therapies for vaginitis may inadvertently disrupt this protective flora. The study aimed to compare the antimicrobial susceptibility profiles of bacterial pathogens isolated from vaginal swabs with those of Lactobacillus strains from vaginal and extra-vaginal sources. Methods: Women with vaginal discharges attending Al-Kafeel Specialized Hospital in Kerbala from March 2023 to April 2024 were enrolled. High vaginal swabs were collected from vaginitis patients. Bacterial isolates were identified using standard culture techniques and the Vitek 2 system. Eight Lactobacillus isolates were obtained from fermented dairy products (n=4), healthy women (n=3), and a probiotic suppository (n=1). Antimicrobial susceptibility testing was performed on bacterial and Lactobacillus isolates using standard methods. A side-by-side comparison of their antimicrobial resistance patterns was conducted. Results: This study of 200 vaginal isolates revealed striking resistance disparities: while pathogenic Staphylococci exhibited &gt;70% macrolide resistance (reaching 90% in Coagulase-negative Staphylococcus) and Streptococci showed 60-83.3% resistance, commensal Lactobacilli maintained 100% susceptibility to levofloxacin, vancomycin and tetracycline, though demonstrating complete resistance to cefoxitin (100%) and high-level aminoglycoside resistance (75-87.5%). Gram-negative pathogens displayed concerning carbapenem resistance (up to 66.7% in Klebsiella pneumoniae), contrasting sharply with Lactobacilli preserved susceptibility profile, highlighting the critical need for microbiome-conscious antibiotic selection in vaginal infections. Conclusions: These findings highlight the potential collateral damage of antimicrobial therapy on vaginal Lactobacilli. Tailored antibiotic selection or probiotic adjuvants may be needed to preserve the vaginal microbiome while treating infections. Keywords: Vaginal microbiota, Antimicrobial susceptibility, Vaginitis, Dysbiosis

The aim of this study was to explore the correlation of ezrin and galectin-3 expressions with prognosis in cervical cancer. The immunohistochemical method was applied to detect ezrin and galectin-3 expressions in normal cervix tissues (n=30), cervicitis tissues (n=28), cervical intraepithelial neoplasia (CIN) tissues (classified as I-III, n=89), and cervical carcinoma tissues (n=84). Follow-up was conducted for 5 to 78 months to analyze the correlation of protein expressions with prognosis. Ezrin and galectin-3 expressions in cervical cancer were significantly higher than in normal cervix, cervicitis and CIN (all P<0.05), and expressions in CIN were significantly higher than in normal cervix and cervicitis (both P<0.05). The expressions of ezrin and galectin-3 were both related with histological grade, deep myometrial invasion and lymph node metastasis (all P<0.05). Spearman analysis showed that ezrin expression was positively correlated with galectin-3 expression in cervical cancer (r=0.355, P<0.05). The survival rate of patients with high expressions of ezrin and galectin-3 was significantly lower than those with low expressions of proteins (both P<0.05). The expressions of ezrin and galectin-3, histological grade, depth of stromal invasion, and lymph node metastasis are risk factors affecting the survival rate of patients with cervical cancer. The expressions of ezrin and galectin-3 were correlated with the development of cervical cancer, and overexpressions of those proteins were indicative of poor prognosis in patients with cervical cancer.

BackgroundIn vitro studies have demonstrated the modulation of vaginal microbiota (VM) by cervical peptides which levels varied with the status of HPV infection and cervical intraepithelial neoplasia (CIN) grades. However, there is a deficiency in population-based studies investigating the modulation of VM compositions and metabolome by cervical differentially expressed genes (DEGs) across different grades of CIN.MethodsThis study included 43 HPV-positive women, classified into low-grade (CIN1, n = 23) and high-grade (CIN2 + , n = 20) groups. Vaginal swabs were collected for both microbiota and metabolome analysis. Cervical exfoliated cells were collected for RNA-Seq analysis.ResultsWe identified 258 differentially expressed genes (DEGs), among which 176 CIN1-enriched genes were linked to immune responses, cell chemotaxis, negative regulation of cell migration, and B cell differentiation, activation, and proliferation. Eighty-two genes upregulated in CIN2 + cohorts were associated with epidermis development and keratinization. Then, we identified 5,686 paired correlations between DEGs, VM, and metabolome, with 2,320 involving Lactobacillus. Further analysis revealed Lactobacillus as the primary determinant of metabolic profiles, followed by Gardnerella, Faecalibacterium, Aerococcus and Streptococcus, such as the notable positive correlation between Lactobacillus with D-lactic acid and DL-indole-3-lactic acid. Applying mediation analysis, we found that Lactobacillus mediated the association of 14 CIN1-enriched DEGs, such as COL4A2, CCBE1 and SPON1, with the production of 57 metabolites, including D-lactic acid, oleic acid and various amino acids. Additional analysis indicated significant mediation effects of 79 metabolites on the association of DEGs with the growth of Lactobacillus, Gardnerella, Fannyhessea and Aerococcus.ConclusionsOur findings provide valuable population-based evidence for the inferred modulation of correlated VM and metabolome by cervical DEGs across different CIN stages.