ObjectiveThe implementation of comprehensive microinvasive cervical cancer (MIC) risk assessment in high-grade squamous intraepithelial lesion (HSIL) patients undergoing loop electrosurgical excision procedure (LEEP) is critical to optimize treatment strategies and improve patient outcomes.MethodsFrom March 2017 to January 2024, a total of 3066 eligible patients with HSIL were retrospectively enrolled from two hospitals and assigned into one training cohort (n = 2084), one internal validation cohort (579) and one external testing cohort (n = 403). Four feature selection methods (Random Forest, Lasso regression, Boruta algorithm, and Extreme Gradient Boosting) were employed to identify key predictive factors from the training cohort. Then, four machine learning models were developed and evaluated using comprehensive metrics. The optimal model was visualized through interpretable techniques and operationalized as a web-based clinical decision support system for real-world implementation.ResultsSix clinical predictive variables were identified, including surgical margins, endocervical curettage (ECC), TCT status, HPV status, Transformation Zone (TZ) type and Age. The optimal model demonstrated good predictive performance, achieving an area under the receiver operating characteristic curve (AUC) of 0.822 (95% CI: 0.793–0.852) in the internal validation cohort and 0.802 (95% CI: 0.730–0.874) in the external validation cohort.ConclusionThe machine learning-based model can accurately assess the risk of MIC during the treatment of HSIL with LEEP, potentially aiding in the selection of appropriate treatment and surveillance strategies in clinical practice.

Cervical cancer is the second leading cause of cancer mortality among Mexican women aged 20-39 years, driven primarily by persistent human papillomavirus (HPV) infection. To determine the prevalence of high-grade squamous intraepithelial lesions (HSIL) in women under 40 years of age and identify associated risk factors. An observational, cross-sectional study was conducted, including 359 women under 40 years old who were evaluated at the Gynecology and Obstetrics Medical Unit of Centro Médico Nacional de Occidente. Cervicovaginal cytology results indicative of HSIL were analyzed to determine prevalence and assess correlations with demographic and gynecological factors. The prevalence of HSIL was 39%, with the highest proportion of cases observed in women aged 35-39 years. The average age of sexual debut was 18.5 years, with an average of 2.93 sexual partners. Women diagnosed with HSIL were more likely to be older, married, and homemakers. HPV infection was highly prevalent across both low-grade squamous intraepithelial lesion and HSIL groups. A high prevalence of HPV infection was found, mostly in a relatively young population. A significant association between infection and risk factors, like marital status and gynecological/obstetric history, was also demonstrated. The findings also confirmed a relationship between HPV and HSIL.

IntroductionThe study of the cervicovaginal microbiome is a critical area of research in medical science. According to scientific data, microorganisms inhabiting the lower female genital tract may influence susceptibility to and persistence of human papillomavirus (HPV), as well as the development and progression of squamous intraepithelial lesions (SIL) toward cervical cancer (CC).MethodsThe study included 67 patients with a histological diagnosis of low-grade squamous intraepithelial lesion (LSIL) and 85 patients with high-grade squamous intraepithelial lesion (HSIL). The cervical canal microbiome of patients with LSIL (n = 31) and HSIL (n = 45) was examined using the 16S rRNA gene sequencing method, while the microbiocenosis of the remaining patients with LSIL (n = 36) and HSIL (n = 40) was analyzed using PCR-RT.ResultsThere are patients with HSIL on average older than patients with LSIL about 3 years. HPV 16 was found to be the most common virus type in patients with SIL. When determining of 16S rRNA genes of prokaryotic microorganisms in women of reproductive age with SIL by the new generation sequencing, it turned out that the cervical canal microbiome is inhabited by many atypical representatives (soil, aquatic and aerobacteria). In addition to the genera Lactobacillus, Streptococcus, Staphylococcus, Gardnerella, Ureaplasma, Prevotella, Sneathia, Fusobacterium, Veillonella, Megasphaera, Dialister, Enterococcus, Escherichia/Shigella, Fannyhessea, Peptococcus, Peptostreptococcus, Finegoldia, Porphyromonas, Corynebacterium, Alloscardovia, Mageeibacillus, Haemophilus, Duncaniella, Ralstonia, Sphingomonas, Pedobacter, Methylobacterium, Ruminococcus, Sediminibacterium, Pseudomonas, Aerococcus, Acinetobacter, Campylobacter, Stenotrophomonas, Sphingobacterium, Phyllobacterium and others may be included in the microbial composition of the cervical canal. Dysbiotic disorders were more common in patients with HSIL. Streptococcus spp. always accompanied aerobic vaginitis (AV), whereas bacterial vaginosis (BV) was associated with the genera Gardnerella, Fannyhessea, Prevotella, Dialister, Sneathia, Anaerococcus, Megasphaera, Finegoldia, Peptoniphilus, Porphyromonas, Parvimonas and Eubacterium spp. When comparing the two methods, the genera Peptoniphilus, Methylobacterium, Ralstonia spp, Sphingomonas, Phyllobacterium, Parvimonas, Anaerococcus and Ruminococcus may be included in the microbial biomass in a significant proportion. Eubacterium spp. did not occur in the 16S sequencing method. Some representatives are detected together with each other in the RT-qPCR method (Femoflor - 16), e.g., Sneathia spp. + Leptotrichia spp. + Fusobacterium spp.; Megasphaera spp. + Veillonella spp. + Dialister spp.; Mobiluncus spp. + Corynebacterium spp., however, Corynebacterium spp., Veillonella spp., Mobiluncus spp., Fusobacterium spp., Leptotrichia spp. and Peptostreptococcus spp. were practically absent (or in a low percentage) in the microbiome of women with SIL.

Combination of photodynamic therapy and estrogen cream for menopause women with cervical low-grade squamous intraepithelial lesion with human papillomavirus infection.

Clinical findings support the critical importance of cervical screening and the management of abnormal cervical cytology results in female organ transplant recipients. We evaluated the incidence and prognosis of atypical cervical lesions in women who are kidney and liver transplant recipients at our organ transplant center. We performed a retrospective cohort study of patients at Başkent University, Ankara, Türkiye, from March 2003 to January 2023. We included women who underwent at least 1 cervical cytology after kidney or liver transplant. The Bethesda System was used to classify cervical cytology results. We analyzed follow-up data by using descriptive statistics. Our study included 124 patients, with 98 (79%) being kidney transplant recipients and 26 (21%) being liver transplant recipients. The median age was 37.5 years. Atypical cervical cytology was detected in 34 patients (27.4%). The distribution of cervical cytology results was 90 with normal cytology (72.6%), 10 with atypical squamous cells of undetermined significance (8.1%), 23 with low-grade squamous intraepithelial lesion (18.5%), and 1 with high-grade squamous intraepithelial lesion (0.8%). Among these, 42 patients (33.9%) underwent colposcopy, revealing abnormal results in 10 patients (23.8%), including 1 (2.38%) with cervical cancer. The findings underline the significant prevalence of atypical cervical lesions among solid-organ recipient patients, highlighting the need for proactive cervical screening and management strategies in this high-risk population.

ABSTRACTPersistent high‐risk human papillomavirus (hHPV) infection, especially HPV‐16, plays a central role in the development of high‐grade squamous intraepithelial lesions (HSIL). This study aimed to evaluate the performance of co‐testing (cytology and hHPV detection) in a real‐world cohort of men who have sex with men (MSM) and transgender women (TW) living with HIV. We conducted a prospective study (2017–2023) at a tertiary care center in Spain. MSM and TW living with HIV underwent screening with anal cytology and PCR‐based hHPV testing. High‐resolution anoscopy (HRA) with biopsy was performed in cases with abnormal cytology and/or hHPV positivity. Clinical, epidemiological, and HIV‐related data were collected to identify risk factors for abnormal cytology and biopsy. 734 cytologies were performed in 380 participants. Abnormal cytology was found in 34%, most commonly ASCUS. hHPV was detected in 63.1% of samples; HPV‐16 was the most prevalent genotype (19.4%), present in 60% of HSIL cytologies and 65% of AIN2–3 biopsies. Risk factors for abnormal cytology included nadir CD4 < 200 cells/μL (aOR 2.61), prior condylomas (aOR 2.66), and infection with any oncogenic HPV genotype (aOR 4.12). Among 91 HRAs, 29.6% showed AIN2–3% and 2.1% in situ carcinoma. HPV‐16, HPV‐52, and HPV‐66 were most frequently associated with abnormal findings in the cytology. In conclusion, anal HPV infection was prevalent in MSM and TW living with HIV. The genotypes most frequently associated with abnormal cytology and histopathological findings were HPV‐16, HPV‐52, and HPV‐66. These findings highlight the potential value of implementing co‐testing strategies in anal dysplasia screening for this high‐risk population.

Safety and feasibility of intramuscular pNGVL4a-CRTE6E7L2 and TA-CIN administration for the treatment of patients with HPV16+ atypical squamous cells of unknown significance or low-grade squamous intraepithelial lesion: A phase I clinical trial

Performance Evaluation of the Aptima hrHPV Nucleic Acid Amplification and Papanicolaou Co-Testing in Cervical Cancer Screening: Insights from a Diverse Population-Based Retrospective Study.

Risk factors for recurrence of high-grade squamous intraepithelial lesion after the two-layer loop electrosurgical excision procedure in patients with positive endocervical, ectocervical and deep margins

GLUT1 Expression Patterns in Verruciform Acanthotic Vulvar Squamous Intraepithelial Neoplasia, HPV-Independent p53 Wild-Type Squamous Cell Carcinoma and Benign Vulvar Lesions.

BackgroundRapid on‐site evaluation (ROSE) of thyroid fine‐needle aspiration biopsy (FNAB) improves diagnostic adequacy and facilitates ancillary molecular testing. In this prospective, multireader study, the authors evaluated the feasibility of using whole‐slide images (WSIs) for ROSE to determine specimen adequacy and preliminary categorization (according to The Bethesda System for Reporting Thyroid Cytopathology [Bethesda]) of image‐guided thyroid FNABs compared with conventional light‐microscopic (LM) examination of the same specimens in a referral cancer center.MethodsThe authors evaluated 98 ultrasound‐guided thyroid FNAB cases. Smears were stained with Papanicolaou and Diff‐Quik and were scanned at ×20 magnification using a Leica Aperio CS2 scanner. Five cytopathologists evaluated specimen adequacy and Bethesda categorization using WSI followed by LM assessment after a 2‐week washout. Intraobserver and interobserver agreements were calculated using Cohen and Fleiss kappa (κ) statistics. Scan time, interpretation time, and the need for ×40 magnification or z stacking were recorded.ResultsIn total, 463 slides were scanned, with mean scan time of 5.48 minutes. WSI quality was acceptable in most cases. Z stacking and ×40 magnification were requested in 23% and 14% of reviews, respectively. Intrareader agreement between WSI and LM examination was excellent (κ = 0.86–0.95). Inter‐reader agreement was moderate for both WSI (κ = 0.48) and LM examination (κ = 0.56). Concordance was highest for Bethesda categories I and VI and lowest for categories III–V. Interpretation with WSI took significantly longer than with LM examination (p < .0001).ConclusionsWSI is a feasible alternative to LM examination for ROSE of thyroid FNABs, with high intrareader agreement and comparable inter‐reader agreement. The limited need for high magnification and z stacking supports its practical utility.

Combining structural equation modeling analysis with machine learning for early malignancy detection in Bethesda Category III thyroid nodules.

Currently, there is no relevant literature specifically discussing the human papillomavirus (HPV) negative conversion rate following Hiporfin photodynamic therapy (PDT) in HPV-associated high-grade squamous intraepithelial lesion (HSIL) of the female lower reproductive tract. Our study aims to fill this gap. Prospective study of 91 patients aged 37.4 ± 13.2 years old with HSIL in the female lower reproductive tract (cervical HSIL: 55, vaginal HSIL: 25, cervical HSIL combined with vaginal HSIL: 8, cervical HSIL combined with vulvar HSIL: 1, vaginal HSIL combined with vulvar HSIL: 2). Hiporfin® (2 mg/kg) was administered intravenously, and 48–72 h later, 630-nm laser irradiation was applied to the lesions. The median follow-up period was 36 months. Before treatment, all patients (91/91, 100%) tested positive for HPV, and only 6 patients (6/91, 6.6%) had been previously vaccinated against HPV. The HPV negative conversion rates were 74.5% (41/55), 89.1% (49/55), 89.8% (44/49) and 95.8% (23/24) at 3–6 months, 12 months, 24 months and 36 months respectively after PDT in treating cervical HSIL. For vaginal HSIL, the negative conversion rates were 28.0% (7/25) at 3–6 months, 52.0% (13/25) at 12 months, 60.9% (14/23) at 24 months, and 64.7% (11/17) at 36 months. In cases of multiple sites of female lower genital tract HSIL, the negative conversion rates were 18.2% (2/11), 45.5% (5/11), 60.0% (6/10), and 62.5% (5/8) at 3–6, 12, 24, and 36 months, respectively. Hiporfin-PDT was most effective in achieving HPV negative conversion for treating cervical HSIL, followed by vaginal HSIL, and finally for multisite female lower genital tract HSIL.

Women treated with cervical loop electrosurgical excision procedure require follow-up to detect residual or recurrent HSIL+, defined as high-grade squamous intraepithelial lesions, adenocarcinoma in situ or cervical cancer. Currently, co-testing with cytology and human papillomavirus (HPV) analysis is usually recommended. This study investigates whether HPV testing alone is comparable to co-testing in detecting HSIL + up to three years after treatment. Recurrence rates of HSIL + are also presented, with follow-up extending up to 18 years. This retrospective cohort study included all 3,540 women treated with a cervical excision in Uppsala County between 1 January 2005 and 31 December 2019. Women with cancer identified in the cone biopsy were excluded. The main outcome was HSIL + detected within three years of follow-up. Sensitivity, specificity and negative predictive value were calculated for the 1,938 women who had a co-testing result as part of their test of cure. Thus, the analysis for the main outcome could finally be performed on 1,938 out of the total number of 3,540 women. Additionally, long-term data on recurrence and time to HSIL+, along with a separate analysis of results prior to cervical cancer diagnosis, were collected for the whole cohort of 3,399 women. The sensitivity and negative predictive value for detecting HSIL + were 69% and 97% for HPV alone, and 74% and 98% for co-testing, respectively. These differences were not statistically significant. Specificity was higher for HPV alone than for co-testing. The negative predictive value of HPV testing for excluding cervical cancer (n = 5) within three years was 100%. Recurrence rate of HSIL + in the three-year follow up was 8%, and the total recurrence rate of HSIL + with a mean follow-up of nine years was 10%. Mean time to recurrence was 28 months. None of 19 cervical cancer cases identified in the long-term follow-up had a co-testing result showing negative HPV but positive cytology. HPV testing alone, as a single test, is comparable to co-testing in detecting HSIL + up to three years after treatment independently of margin status, and demonstrates a higher specificity. Cytology plays a very limited role in the test of cure analysis and could therefore be omitted.

Heterogeneous Distribution of Human Papillomavirus (HPV) Integration Sites in Cervical Precancers Compromises the Diagnostic Accuracy of Integrant-Specific PCR.

Uterine cervical lymphoepithelioma-like carcinoma (LELC) is a rare type of cervical squamous cell carcinoma (SCC). The present study describes a case of cervical LELC, including the cytological findings. A Japanese woman in her 40s was diagnosed with a high-grade squamous intraepithelial lesion (HSIL), suspected cytologically. The patient had previously tested positive for high-risk human papillomavirus (HPV). The cervical biopsy revealed findings that were indicative of HSIL/cervical intraepithelial neoplasia (CIN)2-3, and the patient was then referred to Tokyo Women's Medical University Hospital (Tokyo, Japan). A cervical cytological examination showed HSIL, and the presurgical cervical histological diagnosis was non-keratinizing SCC. Cervical conization was performed and the histopathological diagnosis was LELC. A cervical cytological specimen showed atypical cells with indistinct borders, enlarged round nuclei and a high nucleocytoplasmic ratio. Some cells had prominent nucleoli, and the tumor cells were intermingled with abundant neutrophils and lymphocytes. In addition, atypical squamous cells with orange G-philic cytoplasm and CIN-like atypical cells were observed, as well as clumps of atypical cells with thickened cytoplasm and a metaplastic appearance. The mechanism underlying the contributions of Epstein-Barr virus and/or HPV to the development of LELC is a topic for future investigation.

Cervical high-grade squamous intraepithelial lesion (HSIL) is a well-recognized precursor to cervical squamous cell carcinoma (CSCC). This study aims to explore molecular mechanisms underlying this transition. The datasets of HSIL and CSCC were downloaded from gene expression omnibus database (GEO). HSIL and CSCC coexpression modules were analyzed by Weighted gene coexpression network analysis (WGCNA). Enrichment analysis were analyzed by Gene Ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG). The immune microenvironment (TIME) was analyzed by CIBERSORT. Receiver operating characteristic (ROC) curve, and survival analysis were performed. The binding of FEN1 and its upstream regulator PCNA was determined by protein interaction, correlation analysis and rigid docking. Immune infiltration, immune checkpoint and docking with small molecule inhibitors was also analyzed. In vitro experiments were conducted to investigate the function and underlying mechanisms of FEN1 in HSIL and CSCC. A total of 30 sequentially expressed shared genes were identified as potential drug targets. Immune infiltration revealed an imbalance of M1/M2 macrophages, T γδ cells and T cells CD4 memory resting were negatively associated with tumor progression, forming a pro-cancer effect at the HSIL stage and progressing toward CSCC. The key gene FEN1 was over-expressed in HSIL and CSCC, and correlated with the prognosis of CSCC (P < 0.05). FEN1 interacts with PCNA to regulate macrophage-mediated immune infiltration, increase the risk of immune escape, and decrease the sensitivity to immune checkpoints. Small molecule inhibitor binds to FEN1 docking and inhibits its regulation. FEN1 expression is upregulated in both HSIL and CSCC, with a more pronounced increase in CSCC. Knockdown of FEN1 significantly inhibits the proliferation of cervical cancer cells. This study identifies FEN1 as a biomarker and therapeutic target to truncate or reverse CSCC progression at the HSIL stage.Supplementary InformationThe online version contains supplementary material available at 10.1038/s41598-025-15642-0.

Background: At present, despite numerous excisional methods for the treatment of High-Grade Squamous Intraepithelial Lesion (HSIL), the rates of residual lesions and recurrences remain rather high, ranging from 30% to 50% regardless of the method chosen. Notably, up to 30% of women remain infected with human papillomavirus after surgical treatment. Therefore, the choice of the optimal excision technique for HSIL is still relevant, as it should reduce the incidence of residual lesions, disease recurrence, and human papillomavirus persistence. Aim: The work aimed to evaluate the efficacy of contour-loop excision (C-LETZ) and classical cervical conization in HSIL. Methods: In 2021–2023, at the Krasnopolsky Moscow Regional Research Institute of Obstetrics and Gynecology, 66 patients with histologically confirmed HSIL were examined, treated, and followed up; classical cervical conization (group 1, n = 32) and contour-loop excision of the cervix using a wave-shaped C-LETZ cone (group 2, n = 34) were performed. Results: Positive endocervical resection margins were observed more frequently after classical cervical conization compared with C-LETZ (р 0.001). Conization more often revealed endocervical crypt involvement at the apex of the excised cone (р = 0.03), as well as the presence of HSIL in endocervical curettage (р = 0.01). In group 1, 8% of women had intraepithelial lesions confined to the endocervical curettage with no involvement in the resected specimens, compared with 0% in group 2. Residual lesions in group 1 were detected in 6% (2/32) within one year of follow-up and in 12% (4/32) between one and two years. Disease recurrence after two years of follow-up was diagnosed in 18% of women (6/32). In group 2, residual lesions were identified in 3% (1/34) within one year and in 6% (2/34) between one and two years. Recurrence after two years was three times less frequent compared with group 1—in 6% of women (2/34); p = 0.01. Conclusion: Due to the wave-shaped structure of the C-LETZ loop, excision allows deeper capture of the endocervix, thereby increasing the volume of resected cervical canal tissue. This contributes to a twofold reduction in positive endocervical margins and human papillomavirus persistence, and a 1.5-fold reduction in residual crypt lesions compared with classical conization. Thus, deeper removal of endocervical tissue with C-LETZ reduces the risk of human papillomavirus persistence, residual lesions, and disease recurrence.

To investigate the differential risk of high-grade squamous intraepithelial lesion or worse (HSIL+) associated with single versus co-infection patterns of high-risk human papillomavirus (HR-HPV) genotypes. In this retrospective cohort study, 10,570 patients with abnormal ThinPrep cytology test results and/or HR-HPV infection who underwent colposcopy-guided cervical biopsy at Wuhan Children's Hospital (May 2021-May 2023) were enrolled. Histopathological diagnosis served as the gold standard. Multivariate logistic regression was used to analyze HSIL+ risk across HPV infection patterns, adjusting for age and viral load. Single infections with HPV31, HPV33, or HPV58 demonstrate comparable positivity rates of HSIL+ to HPV16 monoinfection. After adjusting for confounders, logistic regression revealed that co-infection of HPV16 with low-risk HPV genotypes reduced the risk of progression to HSIL+ compared to HPV16 monoinfection (p < 0.05). Similarly, co-infections involving HPV33 or HPV58 (regardless of high/low-risk partners) were associated with lower HSIL+ risk (all p < 0.05). In contrast, HPV31 demonstrated consistent HSIL+ risk irrespective of co-infection status. HPV16, HPV31, HPV33, or HPV58 need equivalent clinical vigilance in screening and management protocols. Co-infection with low-risk HPV genotypes attenuates HSIL+ risk in HPV16-infected individuals, and HPV33/58 co-infections (with any genotype) exhibit protective effects. Our study suggests that HPV31-associated risk might remain unaffected by co-infection, suggesting genotype-specific biological interactions. These findings highlight the importance of genotyping-guided risk stratification in cervical cancer screening.

PurposeThis study aimed to identify reliable risk factors for the development of Vaginal intraepithelial neoplasia (VaIN) within two years after the conization for high-grade squamous intraepithelial lesions (HSIL). We developed a prediction model to predict the risk of VaIN based on preoperative and follow-up data.MethodsWe collected 5358 patients who underwent conization for HSIL, of whom 99 developed VaIN within two years after conization. We selected 495 patients as the control group by randomly pairing them 1:5, and were randomly divided into development and validation cohorts at a ratio 7:3. Random Forest (RF), Lasso, and Extreme Gradient Boosting (XGBoost) were employed to identify the most influential variables in the model development dataset. The optimal variables selected through this process were then used for model construction. Subsequently, four machine learning models were developed, and their performance was evaluated using metrics including sensitivity, specificity, accuracy, area under the curve (AUC), and the F1 score. To enhance interpretability, the prediction process was visualized using Shapley Additive Explanations (SHAP). Finally, the model was deployed as a web-based clinical decision support system for practical clinical applications.ResultsFive key clinical predictive variables were identified: age, transformation zone (TZ) type, presence of VaIN before conization, follow-up cytology after conization, and follow-up HPV after conization. The optimal model demonstrated strong predictive performance, achieving AUC of 0.910 (95% CI: 0.854–0.966) in the internal validation cohort and 0.905 (95% CI: 0.859–0.951) in the external validation cohort.ConclusionWe established a practical and accurate prediction model deployed in the network application to predict the occurrence of VaIN within two years after conization in patients with HSIL. This tool can facilitate targeted clinical decision-making for clinicians.