Low-grade Squamous Intraepithelial Lesion Cases Research Articles

Altered mitochondrial DNA copy number in cervical exfoliated cells among high‑risk HPV‑positive and HPV‑negative women.

The majority of cervical cancer cases are due to human papillomavirus (HPV) infection. However, certain cases of cervical cancer are not caused by HPV. Recent studies have shown a link between altered mitochondrial DNA (mtDNA) copy number, an indicative measure of mitochondrial dysfunction, and cervical cancer in women who test positive for HPV. However, the role of the mtDNA copy number in HPV-negative cervical cancer has remained elusive. In the present study, the mtDNA copy number was determined using quantitative PCR as the ratio between mtDNA and nuclear DNA in 287 ThinPrep cervical samples, including 143 cases with cervical abnormalities and 144 control subjects with high-risk (hr)-HPV positive or HPV-negative status. In an overall analysis of cases categorized based on the cytology diagnosis into squamous cervical carcinoma/high-grade squamous intraepithelial lesions (SCC/HSIL), low-grade squamous intraepithelial lesions (LSIL) and normal controls, the mtDNA copy number was significantly higher in all cases compared to the controls and a higher mtDNA copy number was observed in SCC/HSIL compared to LSIL cases. In the stratification analyses based on hr-HPV positive and HPV-negative status, an increased mtDNA copy number was observed in the cases compared with the controls regardless of their HPV status (P<0.05). When cases with cervical abnormalities were categorized based on histological diagnosis into cervical intraepithelial neoplasia (CIN)2/CIN3 and CIN1, an overall analysis indicated an increased mtDNA copy number in CIN2/CIN3 compared to CIN1 (P=0.01). Stratification analyses of these cases based on HPV status revealed a higher mtDNA copy number in CIN2/CIN3 compared to CIN1 regardless of HPV infection (P<0.05). These results showed that an elevated mtDNA copy number in subjects with cervical abnormalities was not influenced by the HPV status and suggested the possibility of its role in the progression of cervical cancer. The increased mtDNA copy number may be an adaptive response mechanism to compensate for mtDNA oxidative stress and energy deficiency, possibly induced by HPV infection and other environmental exposures.

Detection rate and clinical characteristics of vulvar squamous intraepithelial lesion

Objective: To explore the detection rate, clinical characteristics of vulvar squamous intraepithelial lesion (SIL). Methods: Women diagnosed with vulvar high-grade squamous intraepithelial lesions (HSIL) through colposcopy-guided biopsy from January 1, 2018 to August 31, 2022 in Obstetrics and Gynecology Hospital of Fudan University were included in a 1∶1 ratio with patients diagnosed with vulvar low-grade squamous intraepithelial lesions (LSIL) during the same period. Clinical characteristics including human papillomavirus (HPV) infection rate, genotype, cytology result, colposcopy impression, and lesion location were retrospectively analyzed. Results: (1) The proportion of vulvar SIL detected by colposcopy-guided biopsy increased annually from 2018 to 2022, with rates of 1.64% (740/45 057), 2.34% (1 110/47 402), 2.68% (1 108/41 335), 3.26% (1 536/47 078), 3.31% (667/20 155), with an average rate of 2.57% (5 161/201 027). (2) A total of 1 096 cases of vulvar HSIL and 1 096 cases of vulvar LSIL were included. The overall infection rate of HPV was 92.7% (1 993/2 150), with higher infection rate in vulvar HSIL patients than that in vulvar LSIL patients [96.0% (1 012/1 054) vs 89.5% (981/1 096); χ2=33.62, P<0.001]. Among vulvar HSIL patients, the common HPV genotype from high to low were HPV 16 (66.7%), HPV 52 (14.3%), and HPV 58 (10.0%). For vulvar LSIL patients, the most common HPV genotype were respectively HPV 16 (24.9%), HPV 6 (20.1%) and HPV 52 (17.1%). The overall sensitivity rate of cytology was 53.6%, with no significance difference between vulvar LSIL and HSIL groups (54.3% vs 52.9%; χ2=0.40, P=0.526). The accuracy of colposcopy impression for vulvar HSIL was lower than that for vulvar LSIL [40.2% (163/405) vs 81.7% (380/465); χ2=158.72, P<0.001]. About 57.3% (1 257/2 192) of the patients had concomitant cervical and vaginal lesions, with a higher rate in vulvar HSIL group than that in vulvar LSIL group [62.6% (686/1 096) vs 52.1% (571/1 096); χ2=24.67, P<0.001]. Unifocal lesion was the main type, with no significance difference between vulvar LSIL and HSIL groups [81.4% (381/468) vs 82.5% (386/468); χ2=0.18, P=0.671]. The most common lesion locations were the posterior commissure, followed by labia minora, vaginal vestibule, labia majora, perianal and clitoris. Conclusions: The detection rate of vulvar SIL under colposcopy is about 3%, and the infection rate of HPV is 92.7%. Vulvar SIL, especially vulvar HSIL, is likely to cause concomitant cervical and vaginal lesions. The accuracy of colposcopy in diagnosing vulvar HSIL is low. Therefore a comprehensive and careful examination of the vulva is necessary and suspicious vulvar lesions should be undergone colposcopy-guided biopsy for diagnosis.

Mathematical Modelling of Cervical Precancerous Lesion Grade Risk Scores: Linear Regression Analysis of Cellular Protein Biomarkers and Human Papillomavirus E6/E7 RNA Staining Patterns.

The current practice of determining histologic grade with a single molecular biomarker can facilitate differential diagnosis but cannot predict the risk of lesion progression. Cancer is caused by complex mechanisms, and no single biomarker can both make accurate diagnoses and predict progression risk. Modelling using multiple biomarkers can be used to derive scores for risk prediction. Mathematical models (MMs) may be capable of making predictions from biomarker data. Therefore, this study aimed to develop MM-based scores for predicting the risk of precancerous cervical lesion progression and identifying precancerous lesions in patients in northern Thailand by evaluating the expression of multiple biomarkers. The MMs (Models 1-5) were developed in the test sample set based on patient age range (five categories) and biomarker levels (cortactin, p16INK4A, and Ki-67 by immunohistochemistry [IHC], and HPV E6/E7 ribonucleic acid (RNA) by in situ hybridization [ISH]). The risk scores for the prediction of cervical lesion progression ("risk biomolecules") ranged from 2.56-2.60 in the normal and low-grade squamous intraepithelial lesion (LSIL) cases and from 3.54-3.62 in cases where precancerous lesions were predicted to progress. In Model 4, 23/86 (26.7%) normal and LSIL cases had biomolecule levels that suggested a risk of progression, while 5/86 (5.8%) cases were identified as precancerous lesions. Additionally, histologic grading with a single molecular biomarker did not identify 23 cases with risk, preventing close patient monitoring. These results suggest that biomarker level-based risk scores are useful for predicting the risk of cervical lesion progression and identifying precancerous lesion development. This multiple biomarker-based strategy may ultimately have utility for predicting cancer progression in other contexts.

Clinical Association and Detection of Cervical Cancer with Pap Smear Screening Test

Aim: To find the clinical association of the cervical cancer with pap smear test. Study Design: Prospective study Place and Duration of Study: Department of Obstetrics &amp; Gynaecology, AJK Medical College Muzaffarbad Azad Kashmir from 1st January 2021 to 30th June 2022. Methodology: Five hundred women were screened who were between 20-55 years of age. Sampling was performed by ecto-cervix through wooden Ayre spatula rotation at 360 degrees. The sample was preserved in a formalin jar and sent for glass slide formation. Cytopathological examination presented lesions as negative intraepithelial neoplasia as well as epithelial-cell abnormalities including squamous in addition to glandular cells. Results: The mean age was 41.5±5.6 years. The epithelial abnormalities were diagnosed in 6% of low grade squamous-intraepithelial lesion cases and 1% in high grade squamous intra-epithelial lesion through pap smear. No case of squamous cell carcinoma was identified while 40% showed inflammation presence Most of the females within 31-40 years were having normal cervical morphology while 36%cases inflammation was noticed in 21-30 years of age. Low grade squamous-intraepithelial lesion was noticed in 63.3% of the women within an age group of 14-50 years while high grade squamous intra-epithelial lesion L was observed within 60% of the similar age women. Majority (148.45%) of the women had white discharge from vaginum followed by cervical erosion seen in 19.21% of the cases Conclusion: Pap smear is a highly specific and efficient test for determining pre-cancerous cervical lesions as well as cervical cancers. Key words: Association, Detection, Cervical cancer Pap smear screening test

Classification of Vascular Hotspots and Micro-Vessel Flow Velocity Waveforms in Low-Grade Squamous Intraepithelial Lesions and HPV Condyloma of the Cervix.

To assess hotspot micro-vessel flow velocity waveforms in human papillomavirus (HPV) cervical infections using transvaginal power Doppler ultrasound (TV-PDU) and to explore the associations of these sonographic parameters with HPV condyloma and low-grade squamous intraepithelial lesions (LSIL) of the cervix. A total of 39 patients with cervical HPV infections with abnormal cytology and colposcopy results (26 cases of LSIL; 13 cases of HPV condyloma) were enrolled to assess the vascular classification of the cervix and micro-vessel flow velocity using TV-PDU before treatment; 40 individuals with a pathologically normal cervix were used as the control group; seven parameters were measured, including vascular grading classification (Class I, Class II, and Class III), lowest pulsatility index (PI), resistance index (RI), peak systolic velocity (PS), end-diastolic velocity (ED), time average maximum velocity (TAMV), and the vascular index (VI = PS/ED). According to vascular classification, most LSILs were class I (69.2%, 18/26), followed by class II (26.9%, 7/26) and class III (3.8%, 1/26). Most HPV condylomas were class I (92.3%, 12/13), and one was class II (7.7%, 1/13). PI, RI, VI (p < 0.0001), and the PSs (p < 0.05) were significantly lower in these cases than in the controls. The ED and TAMV were not significantly different between the patients and controls (p = 0.4985 and p = 0.1564). No sonographic parameter was significantly different between LSIL and HPV condyloma. The mean PI, RI, and VI were significantly lower in LSIL than in the controls. For HPV condyloma, a PI of 1.07 had an 84.6% sensitivity, 85.0% specificity, and an AUC of 88.8%; for LSIL, a PI of 1.08 had a 100% sensitivity, 85% specificity, and an AUC of 94.2%; for HPV infection (HPV condyloma + LSIL), a PI of 1.08 had a 94.9% sensitivity, 85% specificity, and an AUC of 92.4%. Hotspot vascular classification and micro-vessel flow velocity waveforms may provide a potential practical method for the auxiliary diagnosis of cervical HPV infection. The PI may represent a valuable index for distinguishing the micro-vessel flow velocity waveforms in LSIL and HPV condyloma. Since the case numbers were limited in the current study, further validation is needed.

Inter- and intraobserver agreement in whole-slide digital ThinPrep samples of low-grade squamous lesions of the cervix uteri with known high-risk HPV status: A multicentric international study.

High-risk human papilloma virus (HR HPV) testing and liquid-based cytology are used for primary cervical screening. Digital cytology, based on whole-slide scanned samples, is a promising technique for teaching and diagnostic purposes. The aim of our study was to evaluate the interobserver and intraobserver variation in low-grade squamous lesions, HR HPV status bias, and the use of whole-slide scanned digital cervical cytology slides. Fifteen expert cytopathologists evaluated 71 digitalized ThinPrep slides (31 atypical squamous cells of undetermined significance [ASC-US], 21 negative for intraepithelial lesion or malignancy, and 19 low-grade squamous intraepithelial lesion cases). HR HPV data were accessible only in the second round. In interobserver analysis, Kendall's coefficient of concordance was 0.52 in the first round and 0.58 in the second round. Fleiss' kappa values were 0.29 in the first round and 0.31 in the second round. In the ASC-US category, Fleiss kappa increased from 0.19 to 0.22 in the second round and the increase was even higher expressed by Kendall's coefficient: from 0.42 to 0.52. In intraobserver analysis, personal scores were higher in the second round. The interobserver and intraobserver variability in low-grade squamous lesions was within fair agreement values in the present study, in line with previous works. The comparison of two rounds showed that expert cytopathologists are generally unbiased by the knowledge of HR HPV data, but that being informed of the HR HPV status leads to a better agreement. Stain quality and back discomfort were highlighted as factors affecting digital cytopathology use.

The role of micronucleus scoring in cervical papanicolaou smears: A 1-year study.

To compare the micronucleus (MN) score in all the major diagnostic categories as per "The Bethesda System for Reporting Cervical Cytology" 2014 including negative for intraepithelial lesions and malignancy (NILM), inflammatory, abnormal squamous cells of undetermined significance (ASC-US), abnormal squamous cells cannot exclude high-grade squamous intraepithelial lesion (HSIL) (ASC-H), low-grade squamous intraepithelial lesion (LSIL), HSIL, and invasive carcinoma (IC) and to assess the role of MN scoring as a biomarker for predicting risk of carcinoma. A total of 1000 conventional cervical smears stained with Papanicolaou (Pap) stain, comprising unsatisfactory for evaluation (86), NILM (140), inflammatory (696), ASC-US (23), ASC-H (16), LSIL (18), HSIL (15), and IC (6) were studied independently by two pathologists, and the number of MN cells per 1000 epithelial cells in high-power (×400) and oil immersion (×1000) was counted and expressed as MN score per 1000 cells. The mean MN score ± standard deviation was found to be 0.99 ± 0.744 in NILM cases, 0.67 ± 0.782 in inflammatory cases, 1.57 ± 0.507 in ASC-US cases, 1.63 ± 0.50 in ASC-H cases, 1.56 ± 0.511 in LSIL cases, 2.47 ± 0.516 in HSIL cases, and 3.0 ± 0.00 in IC cases. A step-wise increase was observed in MN score from inflammatory to IC categories. MN score is a reliable and easy test that can be used in conjunction with routine cervical PAP to assess the risk of malignant transformation in the uterine cervix as a biomarker for predicting the risk of carcinoma.

Hispanic Ethnicity and Cervical Cancer Precursors Among Low-Income Women in Arizona.

PurposeIn the United States, Hispanics are more likely to be diagnosed with cervical cancer compared to Non-Hispanic Whites. Annually, 250,000 to 1 million women are diagnosed with a precursor to CC. The aim of this study was to assess whether Hispanics have a higher prevalence of cervical dysplasia compared to Non-Hispanics Whites among a population of low-income women.Patients and MethodsWe analyzed the results of 10,911 cervical cytology tests administered between 2003 and 2016 that were funded through the Center for Disease Control and Prevention’s (CDC) program for low-income, uninsured women entitled the National Breast and Cervical Cancer Early Detection Program (NBCCEDP). In the state of Arizona, the program is called the Well Women HealthCheck Program (WWHP). Logistic regression was used to identify increased risk of dysplasia, including low-grade squamous intraepithelial lesions (LSIL) and high-grade squamous intraepithelial lesions (HSIL/ICC), and multinomial logistic regression was used to assess increased likelihood for LSIL and HSIL/ICC as separate categories.ResultsIn the crude analysis, Hispanic ethnicity was modestly associated with higher prevalence of LSIL (odds ratio (OR)=1.39, 95% CI=1.01–1.91), but this association was not statistically significant after adjusting for confounders. However, in the final models, lower income was independently associated with LSIL (adjusted odds ratio [aOR]=1.55, 95% CI=1.30–1.44), while smoking (aOR=2.88, 95% CI=1.21–6.84) and no history of Pap test within five years (aOR=3.54, 95% CI=1.61–6.99) were independently associated with HSIL.ConclusionAfter adjusting for confounding in a sample of low-income women with comparable Pap screening rates, ethnicity was not associated with greater prevalence of abnormal pap smears. However, other variables were independently associated with LSIL and HSIL. The higher proportion of LSIL cases among lower income individuals compared to those with higher incomes, and the higher proportion of HSIL cases observed among those screened least regularly stresses the importance of programs like WWHP: programs that target low-income, uninsured women. These programs help save lives.

P16 is superior to Stathmin-1 and HSP27 in identifying cervical dysplasia

BackgroundThe aim of this study was to determine how Stathmin-1 and Heat Shock Protein 27 (HSP27) can be used as adjunctive biomarkers to differentiate high-grade dysplasia from benign/reactive lesions in cervical tissues. In addition, we aimed to see if any of these markers can differentiate endometrial from endocervical adenocarcinomas.MethodsFifty cases including benign cervical tissue, low-grade squamous intraepithelial lesion (LSIL), high-grade squamous intraepithelial lesion (HSIL), adenocarcinoma in situ of the endocervix, invasive endocervical adenocarcinoma, and endometrial adenocarcinoma were selected. Stathmin-1 and HSP27 immunohistochemistry (IHC) were performed for each case and the results were compared to the previously available p16 IHC stains.Resultsp16 stained positively in 100% of HSIL, endocervical adenocarcinoma in situ, and invasive endocervical cases. Stathmin-1 stained positively in 43% of HSIL and 90% of endocervical adenocarcinoma in situ and all invasive endocervical cases. Stathmin-1 and p16 were negative in all benign cervical samples. Stathmin-1, HSP27, and p16 stained 100% of LSIL cases. HSP27 stained indiscriminately, including 100% of benign cervical tissue. 87% of the endometrial adenocarcinomas stained positively for p16, Stathmin-1, and HSP27.Conclusionp16 remains superior to both Stathmin-1 and HSP27 in differentiating dysplasia from benign, reactive changes of the cervix.

Abstract 39: Human Papillomavirus (HPV) DNA Testing Based Cervical Cancer Screening and Triage with CINtec PLUS Cytology Test in HPV Positive Women in Bogalay Township, Ayeyarwady Region, Myanmar

Abstract Purpose: This study aimed to conduct Human Papillomavirus (HPV) DNA testing based cervical cancer screening among married women in Bogalay Township, Ayeyarwady Region, Myanmar and to triage with CINtec PLUS Cytology test among women who have HPV infection. Methods: It was a prospective cross-sectional descriptive study. Total of asymptomatic 677 married women (median age-40-years;range 30-55) residing in Bogalay Township, Ayeyarwady Region, Myanmar were screened in 2020. HPV-DNA testing was performed using cobas HPV-DNA-test which detects 14-high-risk-HPV genotypes -16,-18,-31,-33,-35,-39,-45,-51,-52,-56,-58,-59,-66 and-68. Positive HPV cases were followed by liquid-based- cytology (LBC) and CINtec PLUS Cytology i.e, dual-immuno-biomarker technology to simultaneously detect p-16 and Ki-67 to provide a strong indicator of the presence of transforming HPV infections. Results: HPV was identified in 4.6% (31/677) of screened women in Bogalay Township, Ayeyarwady Region, Myanmar. Among HPV-positive-cases, HPV-16 was 22.6 % (7/31), HPV-18 was 6.5% (2/31) and other-HR-HPV-genotypes (HPV-31,-33,-35,-39,-51,-52,-56,-58,-59,-66and-68) were 70.9% (22/31). In cytological-diagnosis of HPV positive women, 87.1% had cervical cytological abnormalities comprising; 16.1% had high grade squamous intraepithelial lesion (HSIL), 48.4% had low grade squamous intraepithelial lesion (LSIL) and 22.6% had atypical squamous cells of undetermined significance (ASCUS). Only 12.9% were negative for intraepithelial lesion or malignancy (NILM). In CINtec-PLUS-Cytology, all HSIL (100%) cases, 66.7% of LSIL, 14.3% of ASCUS were positive with both p-16 and Ki-67 dual-immuno-biomarker but negative in NILM cases. Conclusion: This study highlighted that using HPV-DNA test in cervical cancer screening, the highest risk women can be determined. In triage with CINtec-PLUS-Cytology in HPV positive women, women who were more significantly at risk for cervical pre-cancer and cancer as well as women that require referral to colposcopy were identified. In LSIL and ASCUS cases, CINtec-PLUS-Cytology may reduce the number of women that require referral to colposcopy. This study pointed out CINtec-PLUS-Cytology was very useful to confirm the high-grade cervical lesions. Citation Format: Mu Mu Shwe, Khin Kay Thwe, Su Su Hlaing, Lynn Pa Pa Aye, Kham Mo Aung, Nu Lwin, Win Maw Tun, Zaw Than Htun. Human Papillomavirus (HPV) DNA Testing Based Cervical Cancer Screening and Triage with CINtec PLUS Cytology Test in HPV Positive Women in Bogalay Township, Ayeyarwady Region, Myanmar [abstract]. In: Proceedings of the 9th Annual Symposium on Global Cancer Research; Global Cancer Research and Control: Looking Back and Charting a Path Forward; 2021 Mar 10-11. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2021;30(7 Suppl):Abstract nr 39.

Comparative Evaluation of SOX2 and p16 Expression in Intraepithelial Neoplasia and Invasive Cancer of Cervix

Introduction: SRY (Sex determining region Y)-Box Transcription Factor 2 (SOX2), a transcription factor functioning as a stem cell marker has been studied in many cancers for its role as an oncogene. This study evaluates the expression of SOX2 and protein 16 (p16) expression in cervical with the intent to establish their role as a diagnostic biomarker. Aim: To evaluate the nature of SOX2 expression in cervical cancer and in intraepithelial lesions of cervix and compare it with the expression of p16 with the intent to establish its role as a diagnostic biomarker. Materials and Methods: This study was a retrospective observational study conducted in the Department of Pathology, Chettinad Hospital and Research Institute, Chennai, Tamil Nadu, India from October 2018 to September 2019. Archival blocks for study were collected from cases between January 2012 to December 2017. Immunohistochemistry for SOX2 and p16 on 61 cases of cervical lesions including SCC, Low-grade Squamous Intraepithelial Lesion (LSIL), High-grade Squamous Intraepithelial Lesion (HSIL) and normal cervix were done. A chi-square analysis was used to determine the relationship of SOX2 and p16 expression in different lesions and compared the same. All collected data was tabulated and analysed by Statistical Package for Social Sciences (SPSS) version 23.0 and was compared by chi-square tests. Results: In the total 61 cases (majority with LSIL, n=21, 34.43%, SCC were 19 (31.15%), HSIL were 20 (32.79%) and adenosquamous carcinoma were 1 (1.64%), SOX2 (p-value &lt;0.001) and p16 (p-value 0.0016) showed over-expression in SCC and HSIL with significant p-value, LSIL showed low expression. SOX2 and p16 expression was limited to the basal one-third in LSIL cases, whereas it was expressed up to two-third or full thickness in HSIL cases. Also, SOX2 and p16 had a significant relationship with p-value=0.001. SOX2 was sensitive for SCC with 84.21% sensitivity and p16 was sensitive for HSIL with 90% sensitivity. Conclusion: Both SOX2 and p16 show increasing expression as the lesion progresses from low grade dysplasia to high grade dysplasia and invasive cancer and can complement each other to make a definitive diagnosis.

Role of Cervical Cancer Biomarkers p16 and Ki67 in Abnormal Cervical Cytological Smear.

Cervical cancer is the most common cancer in India. Screening for cervical cancer helps in marked reduction of invasive cervical cancers. The low sensitivity of Papanicolaou cytology (Pap smear) and high-risk human papillomavirus (HR-HPV) in excluding high-grade intraepithelial lesion (ASC-H) leads to unnecessary referrals to colposcopy-guided biopsy. The combined cervical cytology screening and HR-HPV have its own limitations and still need further standardization. Using additional biomarkers like staining with p16 and Ki-67 might help in triaging abnormal pap smear. A prospective, cross-sectional study was performed over a period of 16 months in the Department of Obstetrics and Gynaecology, in collaboration with Department of Pathology. Study was conducted to know the efficacy of immunostaining with p16/Ki-67 in predicting the presence of significant lesion in cases of mild cytological atypia. PAP smears (conventional and LBC) along with P16, Ki-67 and available biopsies were correlated. Liquid-based cytology (LBC) was done in 2134 cases, out of which 46 cases showed abnormal cytological findings such as [22 atypical squamous cells of undetermined significance (ASCUS), 3 low-grade squamous intraepithelial lesion (LSIL), 8 atypical squamous cells cannot exclude high-grade lesion (ASC-H), 6 high-grade squamous intraepithelial lesion (HSIL), 5 squamous cell carcinoma (SCC), 2 adenocarcinoma, 1 atypical glandular cells of undetermined significance (AGUS)]. Immunostaining with p16 and Ki-67 was performed on 38 cases of abnormal cytological smears. Out of 38 abnormal cytology cases, 28 cases had shown co-staining for both p16 and Ki-67, suggestive of true HPV infection of the cells. Of the 38 cases, 07/14 ASCUS, 06/06 HSIL, 07/08 ASC-H, 05/05 squamous cell carcinoma and 02/02 adenocarcinoma also showed dual positivity for p16 and Ki-67. One case of AGUS was diagnosed, but the smear was unsatisfactory for immunocytochemical evaluation and excluded from the study. Three cases of LSIL were also diagnosed on cytological evaluation, and 1 of them however showed positivity for p16 and Ki-67 on immunocytochemistry (ICC). In the ASC-US group, the sensitivity and specificity of the immunostaining in diagnosing CIN2 + lesions were 87.51%, and in LSIL group, the sensitivity and specificity of the immunostaining in diagnosing CIN2 + lesions were 100%. p16/Ki-67 positivity also increased with cytological severity which in turn corresponded with histological findings: it reached from 50% in ASC-US to 100% in both HSIL and SCC categories. This immunostaining with p16 and Ki67 can be a useful method in the triaging of the ASC-US and the LSIL group as considering the high sensitivity and specificity values.

The value of HPV E6/E7 mRNA quantitative analysis in distinguishing high-grade cervical squamous intraepithelial lesions from low-grade cervical squamous intraepithelial lesions

A quantitative assay for HPV E6/E7 mRNA may be a valuable tool for cervical cancer screening. The purpose of this study is to compare the expression levels of HPV E6/E7 mRNA in high-grade cervical squamous intraepithelial lesion (HSIL) and low-grade squamous intraepithelial lesions (LSIL) and to determine a new method that can be used to distinguish cervical squamous intraepithelial lesions. Routine cytology, HR-HPV E6/E7 mRNA, histology, and p16 immunohistochemistry were performed in tissues from 142 patients with cervical squamous intraepithelial lesions. Significant differences were observed between the E6/E7 mRNA copy number values between the LSIL and HSIL cases (Mann–Whitney U‐test, P < 0.001). The optimal cut-off value (≥9,222.00 copies/mL) was determined using the receiver operating characteristic curve to predict diagnostic performance. Out of the 161 samples tested in this study, four cases were classified cytologically as HSIL but had normal histology. The E6/E7 copy numbers in these cases were all higher than 9,222 copies/mL. Therefore, a quantitative assay for HPV E6/E7 mRNA may be a valuable tool that can be used to distinguish HSIL and LSIL, especially for those with HSIL, for which samples are not obtained by biopsy, or when HSIL is difficult to distinguish by morphology and p16 immunohistochemistry.

An increase of microRNA-16-1 is associated with the high proliferation of squamous intraepithelial lesions in the presence of the integrated state of HR-HPV in liquid cytology samples.

Studies of cervical cancer (CC) have reported that microRNA-16-1 (miR-16-1), which is an oncomiR, is increased in the tissues and cell lines of CC. The aim of the present study was to investigate the association of miRNA-16-1 expression level with squamous cell carcinoma (SCC), the presence of squamous intraepithelial lesions (SIL) and the integration of high-risk human papillomavirus (HR-HPV) DNA. The current study analyzed 80 samples obtained from women by liquid-based cytology, which revealed that 20 were negative for SIL (NSIL) and without HPV, 20 were low-grade SIL (LSIL), 20 were high-grade SIL (HSIL), and 20 were diagnosed as SCC with HR-HPV. The genotyping of the viral DNA was conducted via an INNO-LiPA-HPV array, the expression of miR-16-1 was determined by reverse transcription-quantitative PCR, and the physical state of the HR-HPV was ascertained by in situ hybridization with amplification with tyramide. A total of eight HR-HPV genotypes were distinguished; the most frequent of these being HPV16, followed by multiple infection with HR-HPV (including HPV16). The mixed state of the HR-HPV was observed in 60 and 65% of LSIL and HSIL cases, respectively, while an integrated HR-HPV state was identified in 90% of cases with SCC. The expression level of miR-16-1 increased according to the grade of SIL, and cases with HSIL exhibited a significantly higher miR-16-1 expression level compared with women with NSIL (P<0.001; Table II). It can therefore be determined that the expression of miR-16-1 effects cellular proliferation, due to the viral integration of various HR-HPV genotypes in unique infection or in multiple infection. Thus, the overexpression of miR-16-1 could be monitored in women with LSIL, in order to discard a major lesion.

Cytological Follow-Up of Cervical Intraepithelial Neoplasia 1 after Initial Management During 2-Year Period: Bhumibol Adulyadej Hospital Experience

Objective: To evaluate the frequency of cervical cytological results in patients diagnosed as low-grade squamous intraepithelial lesion (LSIL) in cytology report between women younger than 30 and woman 30 or older after initial management and the completed 2-year follow-up to suggest the proper management for Thai women. Materials and Methods: The present study was retrospectively conducted at Bhumibol Adulyadej Hospital, Bangkok, Thailand between January 2011 and December 2016. The medical records of 279 patients whose cervical cytology reported LSIL were reviewed. The prevalence of silent high grade cervical intraepithelial neoplasia (CIN 2/3) in LSIL cytology result was determined. The results of the cervical cytology after the patients completed the 2-year follow-up were evaluated. Results: During the present study period, 279 cases of LSIL in cytology result were enrolled. In women with LSIL in the cytology report, two-third were in pre-menopausal status. The prevalence of CIN 2/3 in LSIL cytology in women younger than 30 and 30 or older were 11.9% and 11.2%, respectively. At 2-year follow-up, around 97% of the patients who completed the follow-up had a spontaneous regression of the disease in both aged groups. Three percent of cases (3/85) had persistent disease in women 30 or older only. There was no progressive disease in the present study. Conclusion: Silent CIN 2/3 in LSIL cytology in the present study were high. Colposcopy should be recommended for diagnosis and follow-up in this setting. Abnormal Pap at 2-year follow-up was around three percent, therefore, abnormal cytology was still a problem. Continuous close cytology follow-up is still recommended. Keywords: Low-grade squamous intraepithelial lesion, LSIL, Cervical intraepithelial neoplasia, CIN, Cytology

Relationship between cervical lesions and the type-specific viral load of high risk HPV reflected by the Ct value of Cobas 4800 HPV system

Objective: To explore the relationship between cervical lesions and high risk HPV (HR-HPV) viral load reflected by the cycle threshold (Ct) values of Cobas 4800 HPV (Cobas 4800) system. Methods: From August 2016 to September 2017, 7 000 women from Shenzhen, were recruited for cervical cancer screening with Cobas 4800 system and cytology co-test. Colposcope biopsies were performed on women who were positive of HPV 16, 18, and positive of HPV types other than 16,18 with cytology [≥ atypical squamous cell of undetermined signification (ASCUS)], or HPV negative but abnormal of cytology [≥ low grade squamous intraepithelial lesion (LSIL)]. The Ct values of HPV 16, 18 and all combined other types coming from Cobas 4800 system were used as an indicator of viral load to analyze the relationship between type-specific HPV load and the cervical lesions. Results: (1) Among the 7 000 screening women, 370 cases were positive for cervical cancer screening, 325 of them underwent colposcope biopsies, and coloposcopy referred rate was 87.8% (325/370). Among 325 women undergoing cervical biopsy, pathological diagnosis was 119 cases of normal cervical cervix, 151 cases of LSIL, and 55 cases of high-grade squamous intraepithelial lesion (HSIL) and above (HSIL(+); including 53 cases of HSIL, 1 case of cervical adenocarcinoma, and 1 case of cervical squamous cell carcinoma). (2) The Ct value of HPV 16 was inversely correlated with the upgrading of the lesions (r=-0.617, P=0.000), and significant different among normal cervix,LSIL and HSIL(+) (35.4±4.5 vs 31.0±6.0 vs 26.5±4.0; F=25.537, P=0.000). There was no correlation between Ct value of HPV 18 and cervical lesions (r=-0.021, P=0.902). The Ct value of other 12 HPV types was statistically difference among normal normal cervix, HSIL(+) and cervicitis (33.0±5.3 vs 29.9±7.2 vs 29.8±5.8; F=5.087, P=0.007). Among them, LSIL and HSIL(+) were significantly lower than normal cervix (P<0.05), but there was no significant difference between LSIL and HSIL(+) (P>0.05). Conclusion: The Ct value of HPV 16 detecting in Cobas 4800 system as an indicator of virus load obviously correlates with different grades of cervical lesions, therefore could be a reference of cervical lesion existence and an indicator of lesion prognosis.

Follow-up with histopathology and HPV testing on LSIL cytology in China's largest academic woman's hospital.

Cervical cancer remains a major health problem in China. To provide baseline data for establishing the appropriate screening strategy, the authors investigated the histopathologic follow-up and human papillomavirus (HPV) testing results of low-grade squamous intraepithelial lesion (LSIL) cytology in the Chinese Largest Women Hospital. Women with LSIL cytology, human papillomavirus (HPV) testing, and immediate histopathologic follow-up between 2011 and 2016 were analyzed. The frequency of LSIL among 1,095,022 Papanicolaou (Pap) tests was 1.4%, and was significantly higher in cases using liquid-based cytology (LBC; 1.5%-1.6%) compared with conventional Pap smears (CPS; 0.5%). The CPS method had performance for predicting cervical intraepithelial neoplasia (CIN) that was comparable to that of LBC (75% vs 69.1% and 71.9%). Among 8079 LSIL cases with an immediate histopathologic diagnosis, CIN-2/3 or higher (CIN-2/3+) was found in 11.9% of cases and carcinoma in 0.3% of cases. A total of 158 patients (15.7%) with HPV-positive LSIL had a diagnosis of CIN-2/3 compared with 18 patients (5.5%) with HPV-negative LSIL. In postmenopausal women with LSIL cytology, the detection rate for CIN-2/3 in HPV-negative patients was significant lower than that in HPV-positive patients. Furthermore, HPV type 16 (HPV-16) was the most common genotype noted in patients with CIN-2+ lesions, whereas HPV-18 was the most common in patients with CIN-1. Although its reporting rate is lower than that of LBC, CPS is useful due to its low cost and comparable predictive value. HPV testing is helpful for stratifying postmenopausal women with LSIL into lower and higher risk groups. HPV-16 is the most commonly identified genotype in CIN-2+ lesions diagnosed in Chinese women with LSIL cytology, whereas HPV-18 is the most commonly identified genotype in patients with CIN-1 lesions.

Serviksin LSIL, HSIL ve Skuamöz Hücreli Karsinomlarında Biyopsilerde Ki-67, P16, C-erb B2 ve Siklin D1 Ekspresyonunun Önemi

Objective: The squamous cell carcinoma (SCC) of the cervix is the most common malignant tumor of gynecological cancers. Squamous intraepithelial lesions (SILs) are known as a preinvasive lesion. Due to problems in the grading of these lesions that are effective in the development of cervical cancer, immunohistochemical markers are reported to be useful in the differentiation of lesions. In this study, the expression and intensity of the staining pattern of Ki-67, p16, cyclin D1, and c-erb- B2 in HSIL and LSIL cases were investigated.Material and Method: 40 patients with SCC, 40 patients with low grade SIL (LSIL), and 40 patients with high grade SIL (HSIL) who were diagnosed in Department of Pathology of XXX University Medical Faculty between 2003-2011 years were included in this study. C-erb-B2, Ki-67, p16, and cyclin D1 expressions was investigated in the preparations of the patients.Results: C-erb-B2 in 4 cases with SCC (10%), in 6 cases with HSIL (15%), and in 2 cases with LSIL (5%) showed positive staining. There was no statistically significant difference between the three groups (p&amp;gt;0.05). However, in cases with SCC, a strong correlation between c-erb-B2 and the disease stage was observed (p&amp;lt;0.05, r = 0.891). Ki-67 expression was present in average 20% of LSIL cases, average 35% of cases of HSIL, and average 40% of SCC cases. The difference between groups was statistically significant (p&amp;lt;0.05) and due to LSIL. Nuclear staining of cyclin D1 was mainly observed in cases with LSIL, however, cytoplasmic staining was mainly observed in cases with HSIL and SCC. No statistically significant difference between groups was observed with cyclin D1 (p&amp;gt;0.05). p16 expression was found in 39 (97.5%) cases with HSIL, 12 cases (30%) with LSIL. The difference was statistically significant (p&amp;lt;0.05). Conclusion: In conclusion, c-erb-B2 and cyclin D1 expression is not useful in the differentiation of these lesions. However, the Ki-67 may be used to separate LSIL from HSIL and SCC. Also, p16 expression was concluded to be useful in the differentiation of HSIL and LSIL.

An automated quantitative DNA image cytometry system detects abnormal cells in cervical cytology with high sensitivity.

To evaluate the performance of an automated DNA-image-cytometry system as a tool to detect cervical carcinoma. Of 384 liquid-based cervical cytology samples with available biopsy follow-up were analyzed by both the Imager System and a high-risk HPV test (Cobas). The sensitivity and specificity of Imager System for detecting biopsy proven high-grade squamous intraepithelial lesion (HSIL, cervical intraepithelial neoplasia [CIN]2-3) and carcinoma were 89.58% and 56.25%, respectively, compared to 97.22% and 23.33% of HPV test but additional HPV 16/18 genotyping increased the specificity to 69.58%. The sensitivity and specificity of the Imager System for predicting HSIL+ (CIN2-3+) lesions among atypical squamous cells of undetermined significance samples were 80.00% and 70.53%, respectively, compared to 100% and 11.58% of HPV test whilst the HPV 16/18 genotyping increased the specificity to 77.89%. Among atypical squamous cells-cannot exclude HSIL, the sensitivity and specificity of Imager System for predicting HSIL+ (CIN2-3+) lesions upon follow up were 82.86% and 33.33%%, respectively, compared to 97.14% and 4.76% of HPV test and the HPV 16/18 genotyping increased the specificity to 19.05%. Among low-grade squamous intraepithelial lesion cases, the sensitivity and specificity of the Imager System for predicting HSIL+ (CIN2-3+) lesions were 66.67% and 35.71%%, respectively, compared to 66.67% and 29.76% of HPV test while HPV 16/18 genotyping increased the specificity to 79.76%. The overall results of imager and high-risk HPV test agreed in 69.43% (268) of all samples. The automated imager system and HPV 16/18 genotyping can enhance the specificity of detecting HSIL+ (CIN2-3+) lesions.

Presence of koilocytosis in low‐grade smears of high‐risk HPV‐positive women is a negative predictor for cervical intraepithelial neoplasia grade 3 or more

The Netherlands converted to high-risk (hr)HPV-based screening in 2017. An increase in referral of hrHPV-positive women with low risk for cervical intraepithelial neoplasia grade 3 or more (CIN3+) is anticipated and reduction of unjustified referrals will have priority. The relevance of koilocytosis in relation to the underlying risk of high-grade CIN in a primary HPV screening setting is unclear. The aim was to investigate whether the risk for CIN3+ differs between hrHPV-positive atypical squamous cells of undetermined significance (ASC-US)/low-grade squamous intraepithelial lesion (LSIL) with or without koilocytosis. Retrospective cohort study, using data from the Dutch national pathology database (PALGA). The population was 1201 hrHPV-positive women with cytological diagnosis of ASC-US/LSIL. Reporting of koilocytosis was assessed as well as detection rates of CIN1 or less, CIN2 and CIN3+ for ASC-US/LSIL cytology stratified by presence or absence of koilocytosis. Crude and adjusted odds ratios were determined. Koilocytosis was present in 40.1% of ASC-US and 45.9% of LSIL cases. CIN3+ is significantly less often found when koilocytosis is present (7.8% for hrHPV-positive ASC-US with- vs 15.8% without koilocytosis). For hrHPV-positive LSIL this was 11.7% vs 20.2%. The crude and adjusted odds ratios for CIN3+ was 0.45 for hrHPV-positive ASC-US and 0.52 for hrHPV-positive LSIL. The presence of koilocytosis is a negative predictor of CIN3+. The risk of hrHPV-positive ASC-US with koilocytosis is in the same range as hrHPV-positive/cytology negative cases and in a setting of primary hrHPV screening these cases could be followed conservatively by repeat cytology. The results should be confirmed by the first data from the Dutch HPV-based screening programme.