Bethesda System Research Articles

Upregulation of HPV16E1 and E7 expression and FOXO3a mRNA downregulation in high-grade cervical neoplasia.

Cervical cancer remains a significant global health concern, ranking as the fourth most prevalent cancer among women worldwide. Human papillomaviruses (HPV) transcribe many genes that might be responsible for cervical cancer development. This study aims to investigate the correlation between the expression of HPV16 early genes and the mRNA expression of human FOXO3a, a tumor suppressor gene, in association with various stages of cervical precancerous lesions. Eighty-five positive HPV16 DNA cervical swab samples were recruited and categorized based on cytology stages, i.e., negative for intraepithelial lesion or malignancy (NILM), atypical squamous cells of undetermined significance (ASC-US), low-grade squamous intraepithelial lesion (LSIL), atypical squamous cell cannot exclude HSIL (ASC-H), high-grade squamous intraepithelial lesion (HSIL). RT-qPCR was performed to amplify HPV16E1, E4, E6, E6*I, E7, and human FOXO3a mRNA expression in all samples. The relative expression of those genes was calculated using GAPDH as a control. Detection of FOXO3a mRNA expression in the cervical cancer cell line by RT-qPCR and meta-analysis of FOXO3a expression using the RNA-Seq dataset by GEPIA2 were analyzed to support the conclusions. Among the cervical samples, HPV16E1 and E7 were significantly increased expression correlating to disease severity. HPV16E4 mRNA expression was 100% detected in all LSIL samples, with a significant increase observed from normal to LSIL stages. Conversely, FOXO3a mRNA expression decreased with disease severity, and the lowest expression was observed in HSIL/squamous cell carcinoma (SCC) samples. In addition, similar results of FOXO3a downregulation were also found in the cervical cancer cell line and RNA-Seq dataset of cervical cancer samples. HPV16 early mRNA levels, including E1 and E7, increase during cancer progression, and downregulation of FOXO3a mRNA is a characteristic of cervical cancer cells and HSIL/SCC. Additionally, HPV16E4 mRNA expression was consistently detected in all LSIL samples, suggesting the presence of active viral replication. These findings might lead to further investigation into the interplay between HPV gene expression and host cell factors for targeted therapeutic strategies in cervical cancer management.

Value of intraoperative post-conisation human papillomavirus testing in predicting residual or recurrence after treatment with a loop electrosurgical excision procedure in women with HR-HPV positive and cervical high-grade squamous intraepithelial lesion

ObjectiveTo evaluate the feasibility of intraoperative human papillomavirus (IOP-HPV) testing for the prediction of postoperative treatment failure in patients with high-grade squamous intraepithelial lesion (HSIL) undergoing loop electrosurgical excisional procedure (LEEP).MethodsA total of 114 women diagnosed with HSIL by biopsy and/or endocervical curettage who underwent LEEP were included in a prospective cohort study. IOP-HPV testing was performed immediately after the procedure. Patients were followed up for 24 months. Logistic regression was used to analyse the factors influencing the residual or recurrent lesions. Further stratified analyses were performed to investigate the differences in prognosis of IOP-HPV positivity in patients of different age and menopausal status.Results1. Of the 114 patients, 6 (5.26%) were pathologically upgraded to cervical cancer, and 21 (18.42%) were lost to follow-up. Recurrence or residual HSIL lesions occurred in 9.20% (8/87) of cases. Of the 8 women who developed post-treatment HSIL, 7 (26.92%) were positive for IOP-HPV, and only 1 (1.64%) was negative for IOP-HPV (< 0.01).2. Transformation zones of type 2 (P = 0.0306) or type 3 (P = 0.0446), diagnosed as LSIL/negative by cervical biopsy (P = 0.0396), margin involvement (P = 0.0233), positive endocervical curettage after conisation (P = 0.0028), intraoperative HPV-positive (P < 0.01), cytological abnormalities (P = 0.0038), DNA ploidy positivity (P = 0.0172), postoperative HPV (P < 0.01) and DNA ploidy (P = 0.0078) positivity at 6 months were associated with higher risk of residual or recurrent lesions. 3. The results of the multivariate regression analysis showed that IOP-HPV positivity was the independent risk factor for residual or recurrent lesions (OR=10.69 , 95% CI:3.41, 33.51, P<0.01). IOP-HPV positivity was strongly associated with the occurrence of residual/recurrent LSIL (OR=6.42 , 95% CI:1.74, 23.70,P=0.0053) and HSIL (OR=32.08 , 95% CI:3.60, 285.64, P=0.0019).4. Stratified analyses showed that IOP-HPV positive in patients younger than 50 years or premenopausal patients was associated with a significantly higher risk of recurrence or residual lesions (p<0.05).ConclusionsIOP-HPV positivity is an independent risk factor for residual or recurrent HSIL lesions. In addition, IOP-HPV positivity was more associated with residual or recurrent lesions in those younger than 50 years or premenopausal. IOP-HPV testing may be of critical clinical value in providing the early and accurate prediction of residual or recurrent lesions.

Assessing the risk of high-grade squamous intraepithelial lesions (HSIL+) in women with LSIL biopsies: a machine learning-based study

ObjectiveThis study aims to analyze factors associated with the missed diagnosis of high-grade squamous intraepithelial lesions (HSIL+) in patients initially diagnosed with low-grade squamous intraepithelial lesions (LSIL) through colposcopic biopsy and to develop a predictive model for assessing the risk of missed HSIL+.MethodsWe conducted a retrospective analysis of 505 patients who underwent loop electrical excision procedure (LEEP) following an LSIL diagnosis by colposcopic biopsy. Logistic regression was used to identify demographic and pathological parameters associated with missed diagnoses of HSIL+. Additionally, several machine learning methods were employed to construct and assess the performance of the risk prediction models.ResultsThe overall rate of missed diagnoses for HSIL+ was 15.2%. Independent risk factors identified were HPV16/18 infection (OR 2.071; 95% CI 1.039–4.127; p = 0.039), TCT ≥ ASC-H (OR 4.147; 95% CI 1.392–12.355; p = 0.011), TZ3 (OR 1.966; 95% CI 1.003–3.853; p = 0.049) and Colposcopic impression G2 (OR 3.627; 95% CI 1.350–9.743; p = 0.011). Among the models tested, the Decision Tree algorithm demonstrated superior performance with an accuracy of 94.7%, sensitivity of 80.0%, specificity of 96.9%, and an area under the curve (AUC) of 0.936 in the validation set.ConclusionKey independent risk factors for the missed diagnosis of HSIL in patients with LSIL include HPV16/18 infection, TCT ≥ ASC-H, TZ3, and colposcopic impression G2. The Decision Tree model offers a cost-effective, reliable, and clinically valuable tool for accurately predicting the risk of missed diagnosis of HSIL+, facilitating early intervention and management.

Cervical intraepithelial neoplasia and cervical cancer in Hunan Province, China, 2020-2023

ObjectiveTo describe the incidence and distribution of cervical intraepithelial neoplasia (CIN) and cervical cancer (CC) for rural women aged 35-64 in Hunan Province, China, 2020-2023.MethodsData were from the Hunan Provincial Cervical Cancer Screening Program in Rural Areas. Most rural women aged 35-64 in Hunan Province attend the program. All women diagnosed with CINs and CCs will be asked to register detailed information, including pelvic examination, diagnosis, age, education level, etc. CINs included low-grade squamous intraepithelial lesions (LSIL) (CIN1), high-grade squamous intraepithelial lesions (HSIL) (CIN2 and 3); CCs included adenocarcinoma in situ (AIS), early invasive cervical cancer (EICC) (stage Ia1 and Ia2) and invasive cervical cancer (ICC) (stage Ib and above). The incidence of CIN and CC is the number of cases per 1000 women. Chi-square tests (χ2) were used to examine if there were significant differences in proportions among different groups.ResultsA total of 4150700 women were included, and 67071 CINs and CCs were identified. The incidence of LSIL, HSIL, AIS, EICC and ICC were 10.63‰(95%CI: 10.53-10.73), 4.98‰(95%CI: 4.91-5.05), 0.06‰(95%CI: 0.06-0.07), 0.23‰(95%CI: 0.22-0.25), and 0.26‰(95%CI: 0.24-0.27), respectively. The proportion of previous pelvic examinations was relatively low in EICC (17.24%) and ICC (17.45%) (χ2 = 236.57, P &amp;lt;0.01), present abnormal examination was relatively high in AIS (51.33%), EICC (49.74%) and ICC (62.45%) (χ2 = 331.15, P &amp;lt;0.01). HPV16 was the most common high-risk type for LSIL (22.01%), HSIL (36.92%), AIS (46.01%), EICC (58.88%), and ICC (64.34%). The proportion of HSIL was relatively high in women aged 35-44 (27.03%), AIS was relatively high in women aged 45-54 (46.39%), EICC (44.24%), and ICC (48.58%) was relatively high in women aged 55-64. The proportion of ICC was relatively high in women with elementary school (38.68%), HSIL (15.10%) and AIS (17.49%) was relatively high in women with senior high school, AIS (1.52%), EICC (0.62%) and ICC (0.75%) was relatively low in women with university and above. (P &amp;lt;0.01).ConclusionWe have described the incidence and distribution of CIN and CC among rural women aged 35-64. These findings were clinically relevant and were useful for clinical counseling and early diagnosis of CC.

An observational study of clinico-pathological characteristics of gynecological tumors and mode of treatment

Background: Gynecological malignancies are a group of various malignancies of the female reproductive system which included cancers of the ovary, cervix, uterus, vulva, vagina, and also gestational trophoblastic neoplasia. Aims and Objectives: The aim of this study was to study the clinical profile, different histopathological types of gynecological tumors, and mode of treatment. Materials and Methods: Study was conducted on 94 patients from June 2022 to December 2022. Results: Mean age of study population was 49.54±14.54 years. On radiological examination, out of total 52 ovarian cancers, cysts were present in 14 (14.89%) women, solid cysts in 16 (17.02%), papillary projections in 9 (9.57%), vascularity in 11 (11.70%), and lymphadenopathy in 2 (2.12%) women. The Pipelle biopsy results showed complex hyperplasia with atypia in 6 women (6.38%), endometrial carcinoma in 2 women (2.12%), focal atypia in 3 women (3.19%), and simple hyperplasia with atypia in 7 women (7.44%). On Pap smear/liquid based cytology, 1 (1.06%) woman each had atypical squamous cells, HSIL cannot be excluded, atypical squamous cells of undetermined significance, high-grade squamous intraepithelial lesion, and low-grade squamous intraepithelial lesion. No intraepithelial lesion or malignancy was found in 82 (87.23%) women. Present study showed that cervix cases was 10 (10.63%), endometrium 30 (31.91%), and vaginal 1 (1.06%). Majority of women in the present study, that is, 53 (56.38%) had ovarian tumors. In endometrial cancer, endometrioid adenocarcinoma, that is, 15 (50%) followed by simple endometrial hyperplasia without atypia and complex endometrial hyperplasia with atypia, that is, 4 (13.33%) each. Conclusion: Maximum number of patients having various gynecological tumors presented with stage II disease and above but maximum women with ovarian cancer reported with stage III and stage IV. These results clearly explained that ovarian cancer usually had non-specific or misleading symptoms as compared to cervical cancer which reported few specific symptoms.

Perinatal outcomes after surgical treatment of cervical LSIL and conservative treatment of endometrial cancer. A clinical case analysis

This article describes a clinical case of pregnancy management in a patient after surgical treatment of low-grade squamous intraepithelial lesions (LSIL) of the cervix and endometrial cancer. In the structure of female oncological morbidity, precancerous conditions and malignant diseases of the female genital organs occupy leading positions and require the most sparing treatment in women who have not realized reproductive function. The frequency of miscarriage in early pregnancy associated with a history of endometrial cancer, requiring repeated invasive manipulations to assess the effectiveness of treatment, reaches 36–40%, depending on the age of the woman. After surgical interventions on the cervix, regardless of the method of exposure, the frequency of miscarriage significantly increases due to the formation of an organic form of isthmic-cervical insufficiency, which dictates the need for preventive cervical circumcision in a regulated time. In addition, the results of numerous studies also indicate an infectious cause as the main etiological factor in the development and progression of isthmic-cervical insufficiency. Management tactics, obstetric risks, as well as the possibilities of choosing a treatment method are discussed on the basis of the presented clinical case.

Performance of Afirma Genomic Sequencing Classifier in Binary Subcategories of Atypia of Undetermined Significance Thyroid Nodules: Single Versus Repeat Diagnosis.

Background: Afirma Genomic Sequencing Classifier (GSC) testing has been utilized for further risk stratification of thyroid nodules categorized as atypia of undetermined significance (AUS). The 2023 Bethesda system subcategorizes AUS diagnosis into AUS with nuclear atypia (AUS-N) and other atypia (AUS-O). The current study aims to determine if performance of GSC testing differs between the two AUS subcategories and between single AUS cohort and repeat AUS cohort. Methods: This retrospective study analyzed consecutive thyroid nodule fine-needle aspiration with a single or a repeat AUS diagnosis and a diagnostic GSC testing result (benign vs. suspicious). All AUS nodules were divided into AUS-N or AUS-O subcategory and followed by either surgical intervention or at least 12 months of clinical and/or ultrasound monitoring. We then assessed performance of GSC testing in each subcategory and subsequently compared the individual performance in AUS-N or AUS-O subcategory between single AUS cohort and repeat AUS cohort. Results: The study identified a total of 365 thyroid nodules subcategorized as AUS-N (N = 106) and AUS-O (N = 259). Both cohorts showed a significantly lower GSC benign call rate (BCR) in AUS-N nodules compared with AUS-O nodules (43% vs. 71% in single AUS, p = 0.001; 58% vs. 74% in repeat AUS, p = 0.02). The proportion of histology-proven malignancies associated with a suspicious GSC result tended to be greater in AUS-N nodules than AUS-O nodules (28% vs. 10% in single AUS, p = 0.09; 38% vs. 27% in repeat AUS, p = 0.3). Compared with AUS-N nodules, AUS-O cohorts demonstrated significantly higher specificity in the single AUS group (73% vs. 51%, p = 0.01). In both subcategories, the repeat AUS cohort yielded greater specificity, positive predictive value, and diagnostic accuracy compared with the single AUS group. However, the differences did not reach statistical significance. Conclusions: GSC BCR and diagnostic performance of GSC testing may vary in AUS-N versus AUS-O subcategories. However, there were no statistically significant differences in GSC performance between single and repeat AUS cohorts.

2024 Hong Kong College of Obstetricians and Gynaecologists Guidelines for cervical cancer prevention and screening.

Primary prevention of cervical cancer is best achieved by vaccinating girls with a prophylactic human papillomavirus (HPV) vaccine. Despite the high efficacy of such vaccines, cervical cancer screening remains necessary because current vaccines do not offer full protection. Secondary prevention via cervical screening should target all women from age 25 years or at the onset of sexual activity, whichever occurs later, until age 64 years. Screening is recommended at 3-year intervals after two consecutive normal annual cytology results, or at 5-year intervals using HPV-based testing (either HPV co-test with cytology or HPV stand-alone). Women who have undergone hysterectomy with cervix removal for benign disease and have no prior history of cervical dysplasia can discontinue screening. Women with HPV-positive, cytology-negative co-test results should either undergo repeat co-testing in 12 months or immediate HPV16/18 genotyping. Immediate referral of women with positive stand-alone HPV test results for colposcopy without further triage is not recommended. A second triage test using cytology, genotyping for HPV16/18, or p16/Ki-67 dual-stain should be conducted to accurately identify women at high risk for high-grade lesions who thus require colposcopy referral. Women with HPV-positive, cytology-positive co-test results, or high-grade abnormal cytology results should be referred for colposcopy. Treatment with a loop electrosurgical excision procedure is recommended for women with high-grade squamous intraepithelial lesions (HSILs). After HSIL treatment, long-term follow-up with HPV-based testing over 25 years is preferred. When cytology results show atypical glandular cells, colposcopy and sampling of the endocervix and endometrium are recommended.

Diagnostic Performance of Cytocolposcopy versus Biopsy for Premalignant and Malignant Lesions in a Women's Hospital Dysplasia Clinic

Background: Cervical cancer (CRC) is a public health problem because it is the fourth most common gynecologic neoplasm worldwide. The screening tests used to diagnose this pathology are cervical cytology, which in suspected malignancy or with malignancy requires colposcopy to identify the affected area and thus guide the biopsy, which is the gold standard for diagnosis. Therefore, these tests are complementary, and a high diagnostic concordance is required to make a confident diagnosis. Subjects and Methods: A retrospective, cross-sectional, observational, and analytical study was performed. A total of 1470 medical records were analyzed, of which 175 patients met the inclusion criteria. The cyto-colposcopic diagnostic yield was compared with the histopathologic yield. The concordance between screening tests and the gold standard was calculated using Cohen's kappa coefficient Results: The sample comprised 175 subjects who met the selection criteria (11.9%). The mean age was 34.59 + 11.01 years, ranging from 17 to 65 years. The mean sexual debut was 16.6 years, with a mean of 3.1 ± 2 sexual partners. When patients were classified according to lesion type, the highest percentages were found in low-grade squamous intraepithelial lesions (LSIL). With 45.71, 61.14, and 49.14% for cytologic, colposcopic, and histopathologic examination, respectively. The highest concordance between histopathology and cytology was found in the high-grade squamous intraepithelial lesion (HSIL) with 0.41, and the concordance between histopathology and colposcopy in HSIL and cancer was 0.55 and 0.74, respectively. Conclusions: Papanicolaou tests and colposcopy showed moderate concordance with histopathologic findings; the diagnostic accuracy of colposcopy is superior to that of cytology.

Performance of the AmpliSeq NGS Panel in Thyroid Nodules with Indeterminate Cytology.

Fine needle aspiration (FNA) cytological analysis fails to confirm the benignity or malignancy of Bethesda III, IV and V thyroid nodules. Molecular tests performed on FNA samples have demonstrated interesting results in improving the diagnosis of these nodules. The aim of this study was to assess the performance of a large next-generation sequencing (NGS) panel in thyroid nodules with indeterminate cytology (Bethesda III, IV, V). Retrospective, monocentric study including 121 patients with cytologically indeterminate thyroid nodules (Bethesda III, IV and V), which underwent a routine FNA procedure for molecular testing, using the AmpliSeq general cancer NGS panel, with available final histological diagnosis. The main objective was to estimate the negative predictive value (NPV) of malignancy of the AmpliSeq panel in Bethesda III, and IV thyroid nodules. Performance assessment (sensitivity, specificity, positive predictive value (PPV) and NPV) was carried out in the grouped categories III and IV, in the overall cohort, and in each Bethesda category. The final histological diagnosis was used as the designated gold standard. Histologically, 86 nodules were benign and 35 malignant. Molecular analysis yielded a positive result in 40 nodules. Panel performances assessed in the grouped categories Bethesda III and IV, demonstrated a 55.0% (CI95% [31.5;76.9]) sensitivity, a 76.9% (CI95% [66.0;85.7]) specificity, a 37.9% (CI95% [25.7;51.9]) PPV and an 87.0% (CI95% [80.2;91.7]) NPV, considering a 20 % prevalence of malignancy. The performances of the AmpliSeq panel are promising, however, the NPV is not sufficient to avoid diagnostic surgery in cytologically indeterminate thyroid nodules.

Fate of nondiagnostic thyroid fine needle aspirations.

Thyroid nodules may be detected during the workup of thyroid hormone abnormalities and as incidental findings during unrelated imaging studies. The diagnosis of a thyroid nodule is mainly established by performing fine needle aspiration (FNA) under ultrasound guidance. Thyroid nodules are classified as nondiagnostic, defined in the Bethesda System for Reporting Thyroid Cytopathology as samples with excess blood, cyst fluid only, and lack of thyroid follicular cells. The current study evaluates a series of nondiagnostic FNAs to assess whether repeat sampling improves yield and what patient management, and outcomes are after a nondiagnostic FNA. Thyroid FNAs from 2016 to 2023 were retrieved from our institution archives. All cases were performed under ultrasound guidance and with rapid on-site evaluation. Cases were assigned the Bethesda System Category. Nondiagnostic FNAs were further reviewed for repeat FNA procedures, potential molecular testing, or diagnostic resections. In total 3104 thyroid FNAs were reviewed, with 153 (4.9%) being nondiagnostic. Of the 154 FNAs, there were 129 patients with an average age of 60 and a male-to-female ratio of 1:3.2. Of the 130 patients, there were 50 patients who underwent 55 repeat FNAs. Thirty-seven (67%) of the repeats were benign, 13 (24%) were nondiagnostic again, and 5 (9%) were atypia of undetermined significance (AUS). Molecular testing was performed on repeat FNAs diagnosed AUS. Four cases showed no mutations and had a high likelihood of being benign. One case did have an NRAS Q61R mutation, and resection revealed a noninvasive follicular thyroid neoplasm with papillary-like nuclear features. Seventeen (13% of all cases) with nondiagnostic FNA were resected. Twelve (71%) thyroidectomies showed benign adenomatous nodules. The remainder showed incidental papillary thyroid microcarcinoma (0.1 cm), an infarcted follicular adenoma, a noninvasive follicular thyroid neoplasm with papillary-like nuclear features, and metastatic renal cell carcinoma (2×). Thyroid nodules with nondiagnostic cytology are reassuring of being highly likely a benign nodule. Only 5 of the 55 (9%) repeat FNAs yielded abnormalities, with only one of those being truly a follicular neoplasm (confirmed by molecular testing and resection). No primary thyroid malignancies have been identified in follow-up (repeat FNA or surgery). Clinical and ultrasound follow-up may be more appropriate management for nondiagnostic thyroid FNAs.

Cervical Screening Outcomes in Reproductive-Age Women at Peripheral Health Centers

Objective: to ascertain the prevalence of different cervical abnormalities in connection t pap smear results with age distribution of patient. Method: The demographic makeup of the study population was deduced by calculating the percentages of patients in each age group. Numerous diseases were also identified using the results of the Pap smear test, and percentages associated with each diagnosis were computed. This methodology made it possible to thoroughly analyze the participants' diagnoses and age distribution, which revealed trends in cervical health. One hundred participants in all had Pap smears performed as part of the study. Four age groups were established based on the age distribution data gathered: 18–25, 25–35, 35–45, and 45–55 years old. Result: The majority of participants (37%) were found to be between the ages of 25 and 35, while there was a wide variation of participant ages overall. Pap smear tests are crucial for identifying potentially malignant or precancerous diseases, as evidenced by the majority of patients (71%) receiving a diagnosis of cancer or intraepithelial lesions. There was also representation from the following age groups: 18–25 (9%), 35–45 (28%), and 45–55 (4%). Four percent of the cases had atypical squamous cell lesions, two percent had low-grade squamous intraepithelial lesions, and one percent had high-grade lesions. Conclusion: These findings advance customized screening programs and therapies aimed at specific age groups and diagnostic categories as well as our knowledge of cervical health patterns. People who are fertile should definitely get screened for cervical cancer because the age group of 25 to 35 accounts for a substantial portion of diagnoses. The importance of Pap smear testing for early identification and preventive medicine is underscored by the prevalence of intraepithelial lesions or cancer.

Triage performance of DNA methylation for women with high-risk human papillomavirus infection.

DNA methylation is a promising biomarker for cervical cancer screening. This study aimed to validate the triage performance of cytological DNA methylation for detecting cervical intraepithelial neoplasia of grade 3 or worse (CIN3+) in women with high-risk human papillomavirus (hrHPV) infection from a large prospective cohort undergoing opportunistic screening in China (METHY3). The triage performance for detecting CIN3+ lesions was compared between HPV16/18 genotyping, a liquid-based cytology (LBC) test, and the PAX1 and JAM3 methylation (PAX1m/JAM3m) test according to cervical pathologic outcomes. Among the 4394 women infected with hrHPV, 1105 had definitive cervical histological findings that were analyzed. For detecting CIN3+, the specificity of HPV16/18(+), the LBC result of ≥atypical squamous cells of undetermined significance (ASCUS), and PAX1m/JAM3m(+) was 66.4%, 23.9%, and 89.6%, respectively, with odds ratios of 4.24 (95% confidence interval [CI], 2.85-6.40), 4.44 (2.27-10.1), and 18.5 (12.1-28.7) (P < .001), respectively. PAX1m/JAM3m(+) had the highest area under the receiver operating characteristic curve (0.790, 95% CI, 0.747-0.832) in the whole cohort and in women of various ages. PAX1m/JAM3m (+) was detected in all patients with cancer (n = 28). Compared with HPV16/18 genotyping and the LBC test, PAX1m/JAM3m testing reduced referrals to colposcopy by 20.64 percentage points and 61.18 percentage points, respectively. PAX1 m /JAM3 m testing is highly specific for detecting CIN3+. As a triage biomarker, it is superior to HPV 16/18 genotyping and LBC testing for women with hrHPV infection.

Expression of CK17 and SOX2 in Vulvar Intraepithelial Neoplasia: A Comprehensive Analysis of 150 Vulvar Lesions

Background: Recently, the immunohistochemical markers cytokeratin 17 (CK17) and SRY-box2 (SOX2) have been evaluated as adjuncts for the diagnosis of high-grade vulvar intraepithelial neoplasia (VIN). In the present study, the aim was to assess CK17 and SOX2 expression in VIN by studying 150 vulvar lesions, originally reported as high-grade VIN and to assess the diagnostic accuracy. Methods: All slides (H&amp;E, p16INK4a, p53, Ki-67, CK17, and SOX2 stains) were independently assessed by six pathologists and the final diagnosis was reached in consensus meetings, as follows: 46 human papillomavirus (HPV)-independent VIN (including 30 p53 mutant and 16 p53 wild-type lesions), 58 high-grade squamous intraepithelial lesions (HSILs), 4 low-grade SILs (LSILs), 37 non-dysplastic lesions, and 5 lesions where the histology was inconclusive. Results: CK17 positivity was observed in 100% p53 wild-type HPV-independent VIN, compared to 73% p53 mutant HPV-independent VIN, 14% HSILs, 0% LSILs, and 24% non-dysplastic lesions. SOX2 positivity was observed in 13% p53 wild-type HPV-independent VIN, 43% p53 mutant HPV-independent VIN, 2% HSILs, 0% LSILs, and 3% non-dysplastic lesions. The highest diagnostic accuracy (89%) for HPV-independent VIN was obtained when combining p53 and CK17 immunohistochemistry. The addition of SOX2 did not further increase diagnostic accuracy. Conclusion: To conclude, aside from p53, both CK17 and SOX2 can be of value for reaching an accurate diagnosis of HPV-independent VIN.

HPV E6/E7 mRNA screening alone can be used as a screening method for cervical cancer in premenopausal women in China: A retrospective study.

This study aimed to assess the value of a HPV E6/E7 mRNA assay (Aptima® HPV [AHPV]) for primary cervical cancer screening combined with menopausal status. A total of 16 917 women underwent AHPV testing and had complete histopathological results at the Affiliated Hospital of Jining Medical University China between January 1, 2017 and March 31, 2022. We evaluated the performance of different screening strategies and combined strategies, as well as evaluations of different menopausal states. When identifying LSIL+ (includes low- and high-grade squamous intraepithelial lesions and invasive cervical cancer [ICC]), the sensitivity (91.2%) and negative predictive value (NPV; 96.6%) were significantly higher for AHPV than for liquid-based cytology assay (LBC; 33.2% and 84.7% for sensitivity and NPV, respectively). Furthermore, the co-testing strategy (cytology combined with AHPV), when compared with AHPV, achieved a slightly higher sensitivity (93.6% vs. 91.2%, respectively, P < 0.001), a similar specificity (61.3% vs. 62.7%, respectively, P = 0.014), a similar positive predictive value (PPV; 37.5% vs. 37.8%, respectively, P = 0.709) and a similar NPV (97.5% vs. 96.6%, respectively, P = 0.001). Moreover, AHPV (when compared with menopausal women) achieved a higher sensitivity (93.5% vs. 77.7%, respectively, P < 0.001), a higher NPV (97.3% vs. 93.9%, respectively, P < 0.001), a similar PPV (37.8% vs. 37.0%, respectively, P = 0.618) and a slightly lower specificity (60.7% vs. 72.1%, respectively, P < 0.001) in premenopausal women. These results were similar when identifying HSIL+ (includes high-grade squamous intraepithelial lesion and ICC). The present study suggests that initial screening with HPV E6/E7 mRNA testing rather than combined screening is a suitable candidate for cervical cancer screening in China (especially for premenopausal women) based on economic reasons.

Correlation of Cytologic Features With Molecular TestingofIndeterminate Oncocytic (Hürthle Cell) Thyroid Lesions.

Indeterminate oncocytic/Hürthle cell lesions on thyroid cytology are reflexed to molecular testing. This study aims to examine the cytologic characteristics of thyroid fine-needle aspiration (FNA) smears with oncocytes classified as atypia of undetermined significance (AUS) with particular molecular patterns that can aid in determining a more conclusive Bethesda category upfront thus decreasing unnecessary testing and associated costs. Our pathology database was searched for thyroid FNAs with AUS for oncocyte predominance from 2019 to 2022. Sixty six cases that underwent ThyroSeq testing (33 positive and 33 negative) were selected. Two cytopathologists reviewed the smears for cellularity, colloid: cell ratio, artifacts, lymphocytes, microfollicles, and oncocytic nuclear atypia. Molecular results and surgical follow-up were obtained through chart review. Statistical analysis was done using STATA16.1 (StataCorp LLC College Station, TX). Among the 33 ThyroSeq-positive cases, 20 had surgical follow-up. Only 1 case was malignant. Hypercellularity, microfollicular architecture, and oncocytic atypia were associated with a higher incidence of molecular alterations. Five of 66 cases demonstrated microfollicles. Four of these had positive molecular findings, and one was negative. Twenty of 33 (61%) cases with positive molecular findings had no oncocytic nuclear atypia. Of the molecular positive cases with oncocytic atypia that underwent resection, seven of nine (78%) were neoplastic. RAS mutations were the most common finding detected in non-neoplastic and neoplastic lesions. Microfollicles were infrequently seen in nodules with oncocytic predominance; however, those that had microfollicles had high incidence of positive molecular findings. Oncocytic nuclear atypia was present in all the resected neoplastic cases. NRAS and KRAS mutations were the most common molecular abnormalities detected.

Thin-layer cervical sample evaluation: Conventional light microscopy versus digital whole-slide imaging.

Whole-slide imaging (WSI) has been adopted in many fields of pathology for education, quality assurance, and remote diagnostics. In 2021, the College of American Pathologists (CAP) updated guidelines to support pathology laboratories regarding the WSI systems validation process. However, the majority of published literature refers to histopathology rather than cytology. The aim of this study was to compare conventional light microscopy (CLM) and WSI in thin-layer cervical samples evaluation according to CAP guideline. A sample set of 64 thin-layer cervical specimens from women 25-64 years old who participated in cervical cancer screening programs in Tuscany was distributed among five cytologists at Institute for Cancer Research, Prevention and Clinical Network (Florence, Italy) for CLM analysis. After 2 weeks, the corresponding 64 digitally scanned slides were available at several magnifications for WSI evaluation. Substantial/near perfect agreement between CLM and WSI evaluation (0.77 ≥ κ ≥1) was observed for the negative for intraepithelial lesion or malignancy (NILM) class with concordance rates from 83.3% to 100%.Variability in concordance was observed among all the cytologists: 50%-85.7% for low-grade squamous intraepithelial lesion (LSIL), 47.1%-100% for high-grade squamous intraepithelial lesion (HSIL), 50%-100% for atypical glandular cells (AGCs) favors adenocarcinoma (ADK) with moderate/near perfect agreement (0.47 ≥ κ ≥1). Concordance and agreement rates were also variable within the "borderline" cytological categories of atypical squamous cells of undetermined significance (ASC-US), atypical squamous cells cannot exclude an HSIL (ASC-H), and AGCs with lower or not computable kappa for some readers. The overall intralaboratory concordance between CLM and WSI was 92.9% with a near perfect agreement (κ =0.84) for NILM. Substantial agreement (κ ≥0.65) was assessed for LSIL, HSIL/squamous cell carcinoma, AGCs, and ADK categories whereas the agreement was lower (κ ≤0.39) for ASC-US and ASC-H. This study showed an overall substantial/near perfect agreement between CLM and WSI for all the cytological categories except the "borderline" ASC-US and ASC-H. Further progress in cytology WSI systems are needed before introducing it in routine diagnostics.

Correlation of different HPV genotype viral loads and cervical lesions: A retrospective analysis of 1585 cases.

To reduce unnecessary examinations and treatments, an effective detection method for differentiating human papillomavirus (HPV)-positive patients is urgently needed. This study aimed to explore the differences in HPV viral loads across various cervical lesions and identify the optimal cutoff value for high-grade squamous intraepithelial lesions (HSILs). This retrospective study included patients with varying degrees of cervical lesions admitted to a hospital between January 1, 2023, and March 1, 2024. The HPV genotype and viral load were determined using BioPerfectus multiplex real-time assay. The differences in HPV genotype viral loads among cervical lesion classifications were analyzed to identify the most applicable type of viral load. The viral loads of HPV16, HPV31, HPV33, HPV35, and HPV58 were significantly associated with the grade of cervical lesions (p<.05), with the HPV16 group exhibiting the strongest correlation (p<.01). The HPV16 viral load demonstrated good sensitivity (Se) and specificity (Sp) for predicting HSIL (Se=81.52%, Sp=64.13%). The three most prevalent HPV genotypes associated with negative, low-grade squamous intraepithelial lesions (LSILs) and HSILs were HPV16, HPV52, and HPV58. HPV33 exhibited the highest prevalence of HSILs, followed by HPV16. High-risk HPV viral load is associated with cervical lesion classification. HPV16 viral load can effectively differentiate HSIL from LSIL with good Se and Sp.

Risk of High-Grade Cervical Dysplasia in Older Patients With a Positive Human Papilloma Virus Cervical Screening Test

Introduction The Australian National Cervical Screening Program has mandated management algorithms that are uniform across all age groups, but evidence is emerging that perhaps the risk of high-grade squamous intraepithelial lesion (HSIL) may decrease in the postmenopausal period. Objective The aim of the study is to identify if patients ≥50 years of age referred to a tertiary colposcopy service have a different risk of HSIL or greater (+). Materials and Methods This is a retrospective cohort study of 3239 referrals to a hospital colposcopy clinic with a positive human papillomavirus (HPV) cervical screening test between December 2017 and May 2023. Patients were included if they were aged 25–74 years, had a cervix, and completed an outpatient colposcopic assessment. Medical records were analyzed to determine the incidence of HSIL+ for each age group and compared them using χ2 and multivariable regression. Results The overall incidence of invasive carcinoma was 0.93% and HSIL/adenocarcinoma-in-situ was 20.38%. Patients ≥50 years of age were less likely to have HSIL or greater diagnosed than those &lt;30 (p &lt; .0001) or those 30–49 (p &lt; .0001), even after considering referral HPV subtype and reflex cytology (OR = 0.45, 95% CI = 0.34–0.59). The risk of invasive carcinoma in those aged ≥50 was no different than those aged 30–49 (p = .982). Conclusions We observed a lower incidence of HSIL or greater with increasing age even after considering referral HPV subtype and reflex cytology. As further data emerges regarding the risk of high-grade disease in those ≥50 years of age, refinement of the risk-based screening algorithm to account for advanced age may be required.

Hiporfin-photodynamic therapy for high-grade squamous intraepithelial lesions of the endocervical canal in young women

The objective of this prospective study was to assess the efficacy of Hiporfin-photodynamic therapy (PDT) int the treatment of young women diagnosed with high-grade squamous intraepithelial lesions (HSIL) of the endocervical canal. This study included 14 patients aged 30 ± 4.2 years (range 21–37) with HSIL of the cervical canal, treated at our hospital from August 2021 to June 2023. Hiporfin (2 mg/kg) was administered intravenously and laser irradiation with a 630 nm wavelength was performed on the cervical canal and cervical surface 48–72 h later. All patients had been observed for at least 12 months. 64.3% (9/14) were nulliparous. The complete response (CR) rate of cervical canal involvement was 100.0% (14/14) at 3–6 months. Pre-PDT, all patients (14/14) tested positive for high-risk human papilloma virus (HR-HPV). The HPV eradication rate was 64.3% (9/14) at 3–6 months and 85.7% (12/14) at 12 months. No serious adverse effects were observed in patients during or after PDT. No recurrence was noted during the long-time follow-up. 6 patients who planned to get pregnant achieved 6 pregnancies including 1 spontaneous abortion, 2 term pregnancies, and 3 ongoing pregnancies. No fetal loss occurred due to cervical incompetence. Hiporfin-PDT may be a promising, fertility-preserving treatment for HSIL of the endocervical canal in young women.