Introduction
Human papillomavirus (HPV) exhibits a remarkable capability for universal infectivity. Specifically, high-risk human papillomavirus (HR-HPV) stands out as one of the most prevalent infectious agents encountered in clinical practice. This variant of HPV is recognized globally as a significant contributor to the development of cervical precancerous lesions and subsequent cancer. After successfully infiltrating the host, HR-HPV replicates extensively within the mature epithelial basal cells. This replication is particularly concerning when the immune system’s strength wanes, as the virus can then randomly integrate into the host genome, leading to cellular mutations that heighten the risk of cancer. Therefore, a proactive strategy for combating cervical cancer is centered around preventing HPV infections, ensuring timely detection, and effectively blocking the transmission of HR-HPV.
With advancements in the understanding of vaginal microecology, there is a growing interest in this vital female-specific microecosystem. Vaginal microbiology is primarily concerned with the diverse range of microorganisms found in the human vagina. This microbiota encompasses a spectrum of organisms, such as bacteria, fungi, viruses, archaea, and protists. A healthy vagina, especially during childbearing years, has conventionally been viewed as a straightforward ecosystem predominated by rod-shaped Gram-positive bacteria known as lactobacilli. The delicate balance of vaginal microecology plays an essential role in preserving female reproductive health by participating in metabolic processes, constructing mucosal barriers, and modulating immune responses. Disruptions in this balance—such as shifts in the vaginal microbiome, alterations in pH levels, and a reduction in hydrogen peroxide production—can diminish the vagina’s defenses against pathogens, increasing susceptibility to reproductive tract infections.
Emerging research indicates a troubling link between cervical HR-HPV infection and disturbances in vaginal microecology. An abnormal vaginal microbiome may foster HPV persistence and complicate treatment efforts. When the vaginal microenvironment is compromised, it can interfere with local immune functions and amplify HR-HPV expression. This condition inhibits apoptotic processes, allowing HPV to elude immune detection and perpetuate cellular dysregulation. As a result, this escalation can accelerate the progression of cervical intraepithelial lesions, ultimately heightening the risk of cervical cancer without prompt intervention. Despite these findings, the precise relationship between HR-HPV infection and vaginal microecology remains underexplored, particularly in the context of predictive models for assessing infection risks in China. Consequently, this study sought to establish a correlation between vaginal microecological shifts and HR-HPV infection, ultimately aiming to devise a robust prediction model relevant to the disease’s prevention and intervention strategies.
Materials and Methods
Study Participants
A total of 2000 patients, from whom complete data were collected according to strict inclusion and exclusion criteria, were included in this study. Among these, 241 patients were identified as having HR-HPV infections, forming the positive group, while the remaining 1759 patients constituted the negative control group.
The study protocol received approval from the Ethics Committee of Xiangyang Hospital of Integrated Chinese and Western Medicine, ensuring adherence to ethical research standards.
Sample Collection
Patients were positioned lithotomically, and a vaginal speculum was utilized to expose the cervix. Aster obtaining access, vaginal secretions were carefully collected from the lateral wall and posterior fornix using a sterile cotton swab for subsequent vaginal microecology analysis. Additionally, cervical secretions were procured with a cervical brush for comprehensive testing.
Sample Detection
Gram staining techniques were employed to identify the dominant bacterial presence and various forms of vaginitis through microscopic examination. A specialized vaginal secretion detector was also utilized to measure glucuronidase (GUS), β-N-acetylglucosaminosidase (NAG), sialidase (SNA), and leukocyte esterase (LE). In identifying indicators of a healthy vaginal microecology, the primary criteria dictated that dominant bacteria should present as gram-positive large bacilli (Lactobacillus), with no pathogenic microorganisms detected alongside all enzymatic tests yielding negative results. For this study, significant reductions or deficiencies in Lactobacillus, coupled with excessive growth of alternate bacterial strains, were specifically labeled as Lactobacillus deficiency; instances of mixed vaginal infections were excluded from the statistics.
For cervical HPV detection, the polymerase chain reaction (PCR)-reverse point-hybrid method was employed, utilizing a dedicated HPV genotyping test kit to detect multiple HPV types.
Methods and steps
① Extraction of DNA: After eluting the cervical brush, 1.0 mL of the resulting liquid was taken and centrifuged at 13,000 r/min for a duration of 10 minutes. Following this, the supernatant was combined with 100 μL of DNA extract (lysate), mixed vigorously, and subjected to a 10-minute heating period in a boiling water bath. After cooling, it was again centrifuged. Subsequently, 5 μL of this supernatant was introduced into the PCR reaction tube for machine analysis, ensuring both negative and positive controls were included in each assessment.
② Amplification of PCR: The amplification process followed specific temperature cycling protocols with carefully timed intervals to ensure accurate results.
③ Hybridization: This phase involved precise washing and color development of the films according to manufacturer instructions to safeguard the reliability of results.
④ HPV positive control: Apart from the internal control exhibiting color, the HPV16 type site should present with proper coloration as well.
⑤ The results were determined by the presence of blue spots at the IC location, while other sites corresponded to various HPV genotypes.
Statistical Methods
Statistical analyses were conducted using advanced software tools for rigorous data evaluation. The study encompassed a comprehensive statistical analysis framework to express data accurately and determine varying group comparisons. Significant single factors were introduced into a multivariate logistic regression model to identify key predictive factors, allowing for the construction of a robust logistic regression prediction model. A receiver operating characteristic (ROC) curve analysis was also utilized to assess the prediction model’s performance and clinical applicability, ensuring the reliability of the findings.
Results
Related Factors and Vaginal Microecological Test Results
The demographic data of the positive group, which included 241 participants, indicated a median age of 35, with quartiles spanning from 31 to 40 years. Conversely, the negative group comprised 1759 cases with a median age of 36, displaying a similar quartile range. Importantly, no statistically significant age difference was found between these two groups. Analyses of the microecological results revealed no notable differences in vulvovaginal candidiasis (VVC), trichomonas vaginitis (TV), and NAG levels between the groups. However, significant deviations were evident in terms of Lactobacillus deficiency, bacterial vaginitis (BV), aerobic vaginitis (AV), along with other key indicators.
Screening for Risk Factors and Construction of the Logistic Regression Prediction Model
Employing multivariate logistic regression analysis established that parameters such as BV, AV, SNA, LE, GUS, and Lactobacillus deficiency emerged as notable risk factors for HR-HPV infection. A nomogram was subsequently constructed based on these identified predictive factors, offering a visual guide for evaluating HR-HPV infection risks.
Evaluation of the Effectiveness of Different Prediction Models
Three distinct predictive models were operationalized: logistic regression, decision tree, and random forest. Each model scrutinized the significance of six variables influencing HR-HPV infection. Notably, the logistic regression model yielded the highest accuracy among the three models, underscored by its associated AUC value from the ROC curve analysis. Subsequent evaluations reaffirmed the logistic regression model’s precision and robustness in predicting the incidence of HR-HPV infection.
Decision Curve Analysis of the Logistic Regression Model
Applying the clinical decision curve allowed for an in-depth assessment of the predictive model’s accuracy. The outcomes demonstrated the effectiveness and reliability of the logistic regression model in forecasting HR-HPV infection rates across patient demographics.
Discussion
HPV is an exceedingly widespread virus, with over 200 variants identified. Nevertheless, the majority of cervical neoplasia cases arise from persistent sexually transmitted infections linked to oncogenic strains such as HPV-16, HPV-18, and others. This disease represents the third most common cancer among women, contributing significantly to global morbidity and mortality numbers. Furthermore, recent investigations have illuminated the correlation between HPV infection and disruptions in vaginal microecology. Understanding this interplay is essential for developing effective prevention and treatment strategies.
Lactobacillus species are crucial for maintaining a healthy vaginal microbiota in women of reproductive age. These beneficial bacteria generate lactic acid, which serves as a primary antimicrobial agent against numerous pathogens. The presence of Lactobacillus plays a protective role against HPV infections by promoting the clearance of the virus and mitigating cervical lesion occurrences. Moreover, Lactobacillus exhibits properties that may hinder HR-HPV from binding to host cells, highlighting the importance of maintaining a balanced vaginal microbiome in preventing HPV-related health issues.
The study advocates for the significance of establishing predictive models based on identified correlation factors to enhance understanding and prevention of HR-HPV infections. It highlights the essential roles played by Lactobacillus and other variables in maintaining vaginal health and preventing disease progression.
Conclusion
Abbreviations
Human papillomavirus (HPV), high-risk human papillomavirus (HR-HPV), bacterial vaginitis (BV), aerobic vaginitis (AV), glucuronidase (GUS), sialidase (SNA), leukocyte esterase (LE), β-N-acetylglucosaminosidase (NAG), odds ratio (OR), confidence interval (CI), receiver operating characteristic (ROC), vulvovaginal candidiasis (VVC), trichomonas vaginitis (TV), area under the ROC curve (AUC).
Data Sharing Statement
The datasets used and/or analyzed during the current study are available from the corresponding author upon reasonable request.
Ethics Approval and Consent to Participate
The study protocol was approved by the Ethics Committee of Xiangyang Hospital of Integrated Chinese and Western Medicine (Xiangyang Traditional Chinese and Western Medicine Ethics Review No. 20230220-001). Written informed consent was obtained from all participants.
Patient consent for publication
Informed consent was obtained from all patients regarding the publication of the data.
Author Contributions
Funding
Funding for this research was provided by Key projects in the medical and health field of Xiangyang Science and Technology Bureau, Hubei Province (Grant No. 2022YL47A).
Disclosure
The authors declare that they have no competing interests.
References
1. Kong TW, Kim M, Kim YH, et al. High-risk human papillomavirus testing as a primary screening for cervical cancer: position statement by the Korean Society of Obstetrics and Gynecology and the Korean Society of Gynecologic Oncology. J Gynecol Oncol. 2020;31(1):e31. doi:10.3802/jgo.2020.31.e31
2. Sarah L, Sarah A, Thies G, et al. A citizen-science-enabled catalogue of the vaginal microbiome and associated factors. Nature Microbiol. 2023;8(11):2183–2195. doi:10.1038/s41564-023-01500-0
3. Condori S, Ahannach S, Vander Donck L, et al. Recent insights into the vaginal microbiota. Microbiota Heal Dis. 2022;4:e771.
4. Liu H, Zhang F, Cai H, Lv Y, Pi M. The progress in studies on abnormal vaginal microbiota and human papillomavirus infection. Matern Child Health Care China. 2023;38:2315–2318. Chinese. doi:10.19829/j.zgfybj.issn.1001-4411.2023.12.047
5. Usyk M, Zolnik CP, Castle PE, et al. Cervicovaginal microbiome and natural history of HPV in a longitudinal study. PLoS Pathog. 2020;16(3):e1008376. doi:10.1371/journal.ppat.1008376
Introduction
Ah, the world of Human Papillomavirus (HPV) – the unwelcome party crasher at every reproductive health gathering. This little menace has a reputation for infecting everyone, everywhere (it’s basically the celebrity of viruses). But don’t let its fame fool you; high-risk HPV (HR-HPV) is a significant player in the cervical cancer drama. It’s like a bad plot twist—one minute, you’re living your life, and the next, you’re learning about precancerous lesions. Seriously, grab your popcorn, because the real reason you’re here is that it sneaks into your immune system’s “VIP lounge,” integrates into your genome, and messes things up big time. You’d think we’d have better security by now!
Now, here’s where it gets intriguing—our vaginal microbiome! We’re talking about a female-specific superhighway of microorganisms, where lactobacilli rule the dominion like benevolent monarchs. Healthy vaginas are generally known to be home to these little rod-shaped guardians, keeping the bad guys at bay (the perpetrators of unpleasant reproductive surprises). The balance of these microbiota is as crucial as, say, the balance between a comedian’s punchline and the audience’s laughter—mess it up, and you’re in hot water.
Recent studies have suggested that a disarray in vaginal microecology might just leave the door open for HR-HPV invasions, leading to an unwelcome cervical invasion. It’s like if your bouncer had a momentary lapse in judgment, and the HPV gang strolled right in! The study at hand investigates this connection and attempts to establish a predictive model for HR-HPV risk based on the whimsical dance of vaginal microecology. Spoiler alert: it could pave the way for novel prevention strategies. Grab your lab coats; we’re diving deep!
Materials and Methods
Study Participants
Meet our cast of 2,000 patients – sounds like the beginning of a reality TV show, doesn’t it? Out of this crowd, 241 were like, “Hello, HR-HPV!” and 1,759 others managed to dodge that bullet. All under the careful watch of the Xiangyang Hospital Ethics Committee, ensuring no one was auditioning without a consent form.
Sample Collection
The procedure involved patients assuming the lithotomy position—yes, that’s the one that sounds like a fancy yoga pose! With their cervixes on full display (thanks to the trusty vaginal speculum), the researchers took a delicate swab from some strategic locations. Next, they gathered cervical secretions with a brush, not unlike a makeup artist prepping their palette—only this one’s got way more stakes.
Sample Detection
Gram staining was employed to identify the dominant bacteria, while high-tech gadgets detected a cocktail of enzymes. If this sounds like a recipe for a scientific fever dream, you’re not wrong! The groundwork of our investigation into vaginal microbiota’s dominance was laid, capturing whether Lactobacillus was throwing a wild party or behaving itself.
Statistical Methods
Ah yes, the numbers game! Statistical wizardry was executed through IBM SPSS and R. The research team compared various data sets like seasoned poker players at a Vegas table, looking for strong hands—and they found that Lactobacillus seemed to be the kingpin, protecting against the HPV infiltration. Logistic regression models were constructed, equipping the authors with the tools to predict HR-HPV infection potential like seasoned fortune-tellers.
Results
Related Factors and Vaginal Microecological Test Results
In this riveting study, the authors found significant differences in various microbial tests between HPV positive and negative groups—these included the notorious Lactobacillus deficiency and some tricky forms of vaginitis, which could either be the title of a horror movie or an excellent excuse for cancelling plans!
Screening for Risk Factors and Construction of the Logistic Regression Prediction Model
Cutting to the chase, they built a model that highlighted bacterial vaginosis (BV), aerobic vaginitis (AV), and the unmistakable absence of Lactobacillus as risk factors for HR-HPV. Let us embrace this model as the ‘magic 8-ball’ of cervical cancer predictions. Your health could depend on it!
Evaluation of the Effectiveness of Different Prediction Models
The area under the ROC curve (AUC) for the logistic regression model soared at 0.808, which is akin to winning the lottery in statistical terms. Decision tree and random forest models followed but, let’s be honest, this logistic regression model just gave it all the pizzazz!
Discussion
HPV is the heavyweight champion no one wants to face. With a plethora of strains, it can wreak havoc leading to cervical cancer, which, by the way, is no laughing matter—although the statistics might induce a cringe-worthy chuckle. With numbers skyrocketing annually, it appears our defense strategy needs a serious upgrade! Cue the superhero janitors—our Lactobacilli, who smoothly block the baddies and ensure that the vaginal microbiota stays in check. They generate lactic acid, akin to a protective barrier against sexually transmitted infections and help maintain overall reproductive health.
With significant evidence backing the relationship between reduced Lactobacilli and HPV, it’s fundamental for our fight against infection. The crux of the study allows us to appreciate that our vaginal guests play not just a role but an incredibly pivotal one! So, let’s celebrate our Lactobacillus; it deserves a place in the limelight as a protector of our health!
Conclusion
This study shines a spotlight on the entwined fate between HR-HPV and the enigmatic vaginal microbiome. With logistic regression models leading the prediction pack, and the cozy relationship between Lactobacillus and HPV providing a potentially life-saving insight, we find ourselves at the crossroads of opportunity and responsibility. By harnessing this knowledge, we might just conquer the ever-present susceptibility to cervical cancer. Let’s champion our probiotics and prepare for a healthier, HPV-free future!
Abbreviations
HPV, HR-HPV, BV, AV, GUS, SNA, LE, NAG, OR, CI, ROC, VVC, TV, AUC – because in science, we do love our acronyms as much as a good punchline!
Data Sharing Statement
Datasets are available for those seeking the truth—prepare your requests wisely!
Ethics Approval and Consent to Participate
This research played by the rules, as approved by the Ethics Committee, with written consent obtained in accordance to the grand tradition of ethical research.
Patient consent for publication
Every patient said, “Yes, publish away!” – That’s the collaborative spirit!
Author Contributions
Funding
A hearty thank you to the Xiangyang Science and Technology Bureau for providing the treasure chest for this research expedition (Grant No. 2022YL47A).
Disclosure
No conflicts of interest here; just pure science and dedication!
References
References, because knowledge is power!
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