LSIL Cases Research Articles

A Diagnostic Study Using STARD Platform on Liquid-Based Cytology in Cervical Smear and Its Positivity Rate among Females with Abnormal Vaginal Conditions Attending a Tertiary Care Center

Abstract Background: Liquid-based cytology (LBC) is a newer method of preparing cervical cell samples. This technique involves collecting cells in a liquid fixative and preparing and evaluating them. Aim: This study aims to investigate cervical smears prepared using the Ezi-Prep LBC method and analyze the positivity rate for cervical cancer and assess the diagnostic accuracy of LBC in detecting cervical abnormalities among females with abnormal vaginal conditions attending a tertiary care center. Materials and Methods: The present diagnostic study was performed in a tertiary care hospital over a period of 1 year (April 1, 2021–November 30, 2021). A total of 230 women, aged 21–70 years, presented at the gynecology outpatient department for either routine cervical screening or with complaints of vaginal bleeding, discharge, or pelvic pain. Cervical smears were prepared using the Ezi-Prep method and stained with ULTRA-PAP stain. Results: LBC smears are monolayer smears and are without nuclear overlapping with spread over a diameter of 16 mm. Minimum cellular overlapping and crowding were observed in LBC smears. The mean age of the participants was 35.5 ± 5.1 years. The smears were studied by using nine morphological parameters. Conclusion: The majority of the smears were inflammatory smears. A total of 16 cases and three cases were found to be bacterial vaginosis and candidiasis, respectively. In an 8-month study, HSIL was found in two cases, whereas LSIL was found in only one case. The positivity rate was observed to be 0.9% in HSIL cases and 0.4% in LSIL cases, 0.9% ASC-US case, and 0.4% invasive carcinoma case.

The predictive value of serum inflammatory markers for the severity of cervical lesions

ObjectiveExploring the predictive value of NLR, PLR, MLR, and SII for the severity of cervical cancer screening abnormalities in patients.MethodsA retrospective analysis was conducted on the data of 324 patients suspected of cervical lesions due to abnormal TCT and/or HPV in our hospital from January 2023 to December 2023, who underwent colposcopy. The pathological results of colposcopic biopsy confirmed that there were 140 cases of chronic cervicitis, which classified as the group without cervical lesions. The cervical lesion group included 184 cases, including 91 cases of LSIL, 71 cases of HSIL, and 22 cases of cervical cancer. Compared the differences in preoperative peripheral blood NLR, PLR, MLR, and SII among different groups of patients, and evaluated their predictive value for the severity of cervical lesions using Receiver Operating Characteristic (ROC) curves.ResultsThe levels of NLR, PLR, and SII in the group without cervical lesions were lower than those in the group with cervical lesions (p < 0.05), and there was no statistically significant difference in MLR (p > 0.05). The comparison of NLR among LSIL, HSIL, and cervical cancer groups showed statistically significant differences (p < 0.05), while PLR, MLR, and SII showed no statistically significant differences (p > 0.05). The AUC of peripheral blood NLR, PLR, and SII for predicting cervical lesions were 0.569, 0.582, and 0.572, respectively. The optimal cutoff values were 2.3,176.48, and 603.56. The sensitivity and specificity were 38.6% and 73.6%, 28.8% and 85.7%, 37.5% and 76.4%, respectively. At the same time, the joint testing of the three had the highest efficiency, with sensitivity of 69% and specificity of 45%.ConclusionAlthough the peripheral blood NLR, PLR, and SII of the cervical lesions patients were higher than those without cervical lesions in cervical cancer screening abnormal patients, the predictive ROC curve discrimination was low. Therefore, it is not recommended to use preoperative peripheral blood inflammatory markers as markers for cervical cancer screening abnormal patient diversion.

Performance of P16INK4a immunocytochemical stain in facilitating cytology interpretation of HSIL for HPV-positive women aged 50 and above.

Few articles have focused on the cytological misinterpretation of high-grade squamous intraepithelial lesion (HSIL). Due to estrogen deficiency, cervical epithelial cells in postmenopausal women tend to show atrophic change that looks like HSIL on Papanicolaou-stained cytology slides, resulting in a higher rate of cytological misinterpretation. P16INK4a immunocytochemical staining (P16 cytology) can effectively differentiate diseased cells from normal atrophic ones with less dependence on cell morphology. To evaluate the role of P16 cytology in differentiating cytology HSIL from benign atrophy in women aged 50 years and above. Included in this analysis were women in a cervical cancer screening project conducted in central China who tested positive for high-risk human papillomavirus (hr-HPV) and returned back for triage with complete data of primary HPV testing, liquid-based cytology (LBC) analysis, P16 immuno-stained cytology interpretation, and pathology diagnosis. The included patients were grouped by age: ≥50 (1,127 cases) and <50 years (1,430 cases). The accuracy of LBC and P16 cytology in the detection of pathology ≥HSIL was compared between the two groups, and the role of P16 immuno-stain in differentiating benign cervical lesions from cytology ≥HSIL was further analyzed. One hundred sixty-seven women (14.8%; 167/1,127) in the ≥50 group and 255 (17.8%, 255/1,430) in the <50 group were pathologically diagnosed as HSIL (Path-HSIL). LBC [≥Atypical Squamous Cell Of Undetermined Significance (ASCUS)] and P16 cytology (positive) respectively detected 63.9% (163/255) and 90.2% (230/255) of the Path-≥HSIL cases in the <50 group and 74.3% (124/167) and 93.4% (124/167) of the Path-≥HSIL cases in the ≥50 group. LBC matched with pathology in 105 (41.2%) of the 255 Path-≥HSIL cases in the <50 group and 93 (55.7%) of the 167 Path-≥HSIL cases in the ≥50 group. There were five in the <50 group and 14 in the ≥50 group that were Path-≤LSIL cases, which were interpreted by LBC as HSIL, but negative in P16 cytology. P16 cytology facilitates differentiation of Path-≤LSIL from LBC-≥HSIL for women 50 years of age and above. It can be used in the lower-resource areas, where qualified cytologists are insufficient, as the secondary screening test for women aged ≥50 to avoid unnecessary biopsies and misinterpretation of LBC primary or secondary screening.

Application of HPV-16 in Liquid-Based thin Layer Cytology of Host Genetic Lesions Based on AI Diagnostic Technology Presentation of Liquid

Nowadays, with the continuous improvement of people's health awareness and the increasing incidence of cancer year by year, the screening work of various clinical diseases, especially cancer, is also widely promoted. Cervical lesion screening is a common type. Cervical lesion screening is used to judge whether women have cervical lesions and increase attention to cervical lesions[1]. Proper prevention, diagnosis and treatment play an important role in ensuring women's health and life safety, and have been widely promoted in many places in our country. At the same time, in the screening of cervical lesions, how to adopt a more efficient and accurate diagnosis is very important to improve the screening effect of cervical lesions. With the rapid development of science and technology, AI and other advanced technologies have been gradually introduced in clinical disease diagnosis, especially in the liquid-based thin-layer cytology examination[2]. To further analyze the application effect of liquid based thin layer cytology AI diagnostic technology in the pathological diagnosis of cervical lesions. In this paper, we investigate the occurrence of HPV-16 integrated human host genome in cervical cancer and precancerous lesions and its application in cervical cancer screening. 50 cases of cervical cancer, 127 cases of LSIL, 83 cases of HSIL were collected, in addition, 22 cases of normal cervical epithelial tissues extracted by total hysterectomy were collected. The integration status of HPV-16 infection was detected by overlapping PCR.

Cervical HPV Infections, Sexually Transmitted Bacterial Pathogens and Cytology Findings-A Molecular Epidemiology Study.

Prevalent cervical HPV infection and high-risk HPV persistence consequences have been extensively investigated in the literature; nevertheless, any causative interrelations of other sexually transmitted bacterial infections (STIs) with cervical HPV infection have not yet been fully elucidated. This study aimed to investigate the possible association of STIs with cervical cytology aberrations and HPV genotyping results in a representative sample of predominantly young Greek women. Liquid-based cytology and molecular detection for bacterial STIs and HPV as well as extended HPV genotyping were simultaneously assessed in cervical samples from 2256 individuals visiting several urban outpatient Gynecology Departments for well-woman visits or cervical screening throughout a 20-month period. All specimens were centrally processed with validated molecular assays. The mean age of the studied women was 37.0 ± 11.7 years; 722 women (33.30%) tested positive for STI (mean age 34.23 ± 10.87 years). A higher mean age (38.34 ± 11.83 years (p < 0.05)) was associated with negative STI testing. Chlamydia trachomatis was detected in 59 individuals (8.2%), Mycoplasma hominis in 156 (21.6%), Mycoplasma genitalium in 14 (1.9%), and Ureaplasma spp. in 555 (76.9%); infections with two bacterial pathogens were identified in 73 samples (10.1%). Cervical HPV was detected in 357 out of 1385 samples with a valid HPV typing result (25.8%). The mean age of HPV-positive women was 32.0 ± 8.4 years; individuals testing HPV-negative were slightly older (N = 1028): 34.4 ± 9.2 (p < 0.05). Among the 1371 individuals with valid results both for bacterial STIs and cervical HPV detection, women with an HPV-positive sample were more likely to harbor an STI (OR: 2.69, 95% CI 2.10-3.46, p < 0.05). Interestingly, bacterial STI positivity illustrated significant heterogeneity between NILM and LSIL cases, with 28.88% of NILM and 46.33% of LSIL cases harboring an STI, respectively (p < 0.05). In brief, in a population with a high prevalence for STIs, especially Ureaplasma spp., an association was documented between bacterial pathogen detection and cervical HPV infection, as well as abnormal cytology; these findings merit further investigation.

Elevated expression of endocan in the development of cervical squamous neoplasia of the uterus.

Accumulated evidence has shown that endocan, which was originally called endothelial cell-specific molecule-1, is an attractive prognostic factor in a variety of cancers. However, the relevance of endocan expression in human malignancies remains to be clarified. In the present study, the expression of endocan in cervical squamous neoplasia of the uterus, including low- and high-grade squamous intraepithelial lesions (LSIL and HSIL, respectively), as well as in invasive squamous cell carcinoma was examined by immunohistochemistry. Endocan was not sufficiently expressed in the normal cervical epithelium. Endocan expression was present in LSIL cases but was limited to basal and parabasal areas of the cells. HSIL cases exhibited strong expression of endocan with widely distributed expression toward the epithelial surface. In contrast, further strong expression of endocan was not observed in patients with invasive carcinoma. This study is the first study showing increased expression of endocan in precancerous dysplastic lesions and malignancy of the cervix. The data suggest that a high expression level of endocan potentially contributes to the development of cervical squamous neoplasia of the uterus.

Association of MicroRNA-10b Expression and P16 with Pathological Features in Various Stages of Cervical Precancerous Lesions.

Cervical cancer is the fourth most prevalent cancer for females with 14,100 new cases each year globally. Efficient screening and intervention at the precancerous stage is the key point to the prevention and treatment of cervical cancer. However, no widely recognized biomarkers have been discovered yet. We investigated the expression of miR-10b in cervical cells and its correlation with clinicopathological features in different pathological grades of cervical precancerous lesions. The expression of miR-10b in cervical cytology samples from 20 cases of LSIL, 22 cases of HSIL, 18 cases of early-stage cervical cancer, and 20 cases of cervicitis controls were assessed using qPCR. From the same cervical cytology samples, the human papillomavirus (HPV) load was assessed using semi-PCR and the lesion size, and gland involvement levels from the same subjects were assessed during the cervical examination. The correlation between miR-10b expression and different pathological grades of cervical lesions was analyzed. We also calculated the correlation between HPV load, lesion size, gland involvement, P16 expression, and different pathological grades. The expression of miR-10b exhibited a step-decreasing manner from cervicitis control (4.23(4.00,4.71)) to LSIL (2.67(2.52,2.90)), HSIL (1.49(1.30,1.80)) and cervical cancer group (0.65(0.55,0.80)). There is a significant difference (P<0.001) between cervicitis and HSIL, cervicitis and cervical cancer, ISIL and HSIL, as well as ISIL and cervical cancer but not between the cervicitis group and the LSIL group. In addition, more severe pathological grades were correlated with a bigger rate of gland involvement (P＜0.001). We also found that different pathological grades were correlated with the intensity of P16 expression (P=0.001), and the intensity of P16 expression is positively correlated with different pathological grades (P<0.05). Repressed expression of miR-10b is related to the progression of cervical precancerous lesions. Increased gland involvement rate and increased intensity of P16 expression are risk factors for developing cervical cancers. Our result showed that miR-10b may be a potential biomarker for the screening and ranking of cervical precancerous lesions.

Comparison of Ki67 index and P16 expression in different grades of cervical squamous intraepithelial lesions.

the assessment of P16 expression and Ki-67 proliferative index is now proposed as an adjunct test for the diagnosis of high-risk precursor lesions for cervical cancer. The aim of the present study was to elucidate the quality expression of P16 and quantification Ki-67 index in different types of cervical intraepithelial neoplasia and also to determine the cutoff for Ki67 index to predict the severity of CIN lesions. This cross-sectional study was conducted on patients with colposcopic indication. Selected samples with different CIN grades were examined for P16 and Ki-67 index by immunohistochemical (IHC) methods. All LSIL (CIN I) cases were negative for P16, while in 58.7% of HSIL cases (CIN 2/3), P16 was positive. The mean Ki67 index in the present study was 3.13 ± 2.65 in the upper two/third of the squamous epithelium in the LSIL group and 19.04 ±36.40 in the HSIL group, which was statistically significant. Also, the mean Ki67 index in full thickness squamous epithelium in HSIL group was significantly higher than LSIL. The sensitivity of P16 and Ki67 index in our study was 58.73%, 66.67% and the specificity was 100% and 100%, respectively. Assessment of P16 expression and Ki67 index can be used to distinguish low grade (CIN1) intraepithelial lesion from high grade (CIN2/3) intraepithelial or precancerous lesions.

Tumor-associated tissue eosinophilia in preinvasive and invasive squamous cell carcinoma of cervix

Background: Differentiating between preinvasive and invasive squamous neoplasms of the cervix can be a challenge in small, superficial fragments of cervical biopsy, particularly when inflammatory infiltrates obscure the epithelial stroma interface and due to tangential sectioning of the acanthotic squamous epithelium. This study evaluates whether thresholds of eosinophils in stroma serve as a morphological marker for the presence of tumor invasion in squamous neoplasms of the cervix. Material and methods: Sixty-five cases of invasive squamous cell carcinoma and preinvasive squamous neoplasia were evaluated. In each case, the number of eosinophils per HPF and per 10 HPF in the tissue adjacent to the neoplastic epithelium was counted. For statistical purposes, the elevated eosinophils were categorized as &gt; 3 eosinophils/HPF, &gt; 5 eosinophils/HPF, and &gt;10/10 HPF. Results: Stromal eosinophils were present in 1/5 cases of LSIL, 3/8 HSIL cases, 15/16 cases of microinvasive squamous cell carcinoma, and 34/36 cases of invasive squamous cell carcinoma, signifying an increase in eosinophil count from a spectrum of preinvasive to invasive carcinoma. Greater than 5 eosinophils/HPF and &gt;10 eosinophils/10 HPF had sensitivity, specificity, and positive predictive values of 55%, 100%, 100%, and 75%, 100%and 100%, respectively. The difference of &gt;5eosinophils/HPF and &gt;10eosinophils/10 HPF between SIL and SCC was statistically significant. Conclusions: Our study suggests that the presence of increased eosinophils in squamous neoplasia of the cervix can be associated with tumor invasion and should prompt a thorough evaluation when evidence of invasion is absent.

Diagnostic Accuracy of p16 Expression in Differentiating Cervical Intraepithelial Neoplasia from Invasive Squamous Cell Carcinoma: A Descriptive Analysis at a Tertiary Care Hospital of Bangladesh

Background: Cervical cancer is one of the most common cancers that affect women worldwide. It is well established thathigh-risk human papillomavirus (HR-HPV) is the prime risk factor for the development of cervical cancer. Early and accuratediagnosis of cervical neoplasia is of utmost significance for prolongation of patient survival. A panel of immunomarkers hasbeen developed and tested to overcome the limitations to histopathological diagnosis. Among them p16 is one of the commonlyused immunomarkers now-a-days. In recent years, routine haematoxylin and eosin (H &amp; E) stain coupled with p16immunohistochemistry (IHC) have a major impact in the field of uterine cervical pathology and cancer screening. Objectives: The present study was aimed to determine the p16 scores and its association with histological types, and grades ofISCC including diagnostic accuracy in differentiating ISCC from CIN lesions. Materials and Methods: This descriptive type of cross-sectional study was conducted in the Department of Pathology,Rajshahi Medical College (RMC), Rajshahi, in collaboration with the Department of Pathology, Bangabandhu Sheikh MujibMedical University (BSMMU), Dhaka, during a period of two years from January 2019 to December 2020 by using routine H&amp; E staining and p16 IHC analysis to evaluate the expression of p16 in samples of cervical biopsies from 51 patients withhistopathologically confirmed diagnosed cases of CIN and ISCC, considering the percentage of p16 positive cells and thereaction intensity. Results: This study revealed that the aberrant expression of p16 increased from LSIL to ISCC, thus emphasizing its usefulnessas an adjunct marker for predicting risk of developing cervical cancer in the test patients. Most of the cases of ISCC in this studyexpressed high level of p16, while only one patient with LSIL failed to express the marker. Notably, 90% of LSIL cases werelow to moderate p16 positive, 100% of HSIL cases were moderate to high p16 positive, while most of the ISCC cases (81%) hadhigh p16 expression. Among p16 positive cases, this is an attempt to verify direct association between lesion severity andreaction intensity. The frequency of positive cells and the reaction intensity were statistically significant (P&lt;0.001) whencompared among different histologic types. Most importantly, the present results clearly demonstrated that p16 IHC wascapable of differentiating ISCC from CIN cases. Conclusion: In addition to routine H &amp; E staining and histopathological diagnosis of cervical lesions, p16 immunomarkercould be used for risk assessment of histologically detected lesions and for distinguishing between ISCC and CIN specimenswhich, in turn, could help predict the progression of cervical lesions and thus monitor the screening of cervical cancer in thecommunity.

High-sensitivity C-reactive Protein and Regression of Low-grade Squamous Intraepithelial Lesion: The Role of Low-grade Inflammation in Cervical Carcinogenesis.

BackgroundInflammation is emerging as a potential mechanism of cervical carcinogenesis. However, few studies have investigated the association between host inflammatory status and the natural course of cervical precursor lesion. The aim of this study was to assess the probability of LSIL regression, associated with an inflammatory biomarker, high-sensitivity C-reactive protein (hs-CRP).MethodsIn a longitudinal cohort study, female participants were examined annually or biannually using cervical cytology between 2006 and 2015. Incident LSIL cases were included in the analysis, with regression defined as at least one consecutive normal cytologic result. A total of 520 women aged 22–64 years were followed up for LSIL regression. The multivariable-adjusted hazard ratios (HRs) for LSIL regression were estimated using a parametric proportional hazards model.ResultsDuring 827.5 person-years of follow-up, 486 out of 520 subjects (93.5%) showed LSIL regression. After adjusting several important potential confounders, a higher quartile of hs-CRP levels was significantly associated with a lower rate of regression (for quartile 4 vs quartile 1, inverse HR 1.33; 95% CI, 1.04–1.69; P for trend = 0.028).ConclusionsThe low rate of spontaneous regression recorded in women with higher hs-CRP lends support to the role of the perturbated host inflammatory status in cervical carcinogenesis, and suggests that hs-CRP level could help monitor LSIL.

Immunohistochemical analysis of CD68, CD4, CD8 and CD20 expression in cervical dysplasia and its relationship with HR-HPV infection.

The aim of the study was to examine the composition of the inflammatory infiltrates in cervical premalignant lesions and contribute to a better understanding of immune response to HR-HPV infection and dysplasia. Semi-quantitative analysis of CD68, CD4, CD8 and CD20 immunohistochemical expression in a series of 41 cervical biopsies without dysplasia, 24 cases of LSIL and 35 HSIL cases was performed. In each subject, genotyping for 12 HR-HPV types was done prior to the biopsy. Observing the total sample, no correlation between CD68, CD4, CD8 and CD20 expression levels and HR-HPV infection was found, regardless of the presence of mono- or co-infection (p>0.05). A statistically significant correlation between dysplastic changes and CD68 expression, as well as between dysplastic changes and CD4 expression, was observed (p=0.003 and p=0.016, respectively). For CD68 expression, there was a positive correlation with both LSIL and HSIL, and concerning CD4 expression, there was a positive correlation primarily with LSIL. The finding of mild CD68 expression shows a 10.5 times greater chance of the sample being classified as LSIL, while the finding of a strong CD68 expression shows a 12 times greater chance of the sample being classified as HSIL, in comparison to cases with no expression. When the samples were stratified in relation to the lesion grade, a correlation between HR-HPV infection and CD68/CD4 expression again was not proved (p>0.05). No correlation between CD8 and CD20 expression with dysplasia was found (p>0.05). We consider a higher prevalence of macrophages and CD4 lymphocytes in dysplastic lesions to be a response to dysplasia rather than HR-HPV infection itself. The increase of the expression levels of macrophages with the degree of the lesion speaks in favour of their potential role in the progression of the neoplastic process.

To develop cytology procedure as a routine and to get familiar with cytological appearances of the commonly encountered cervical and vaginal lesions

The material for the current investigation contained assessment of 1542 cervical/vaginal smear, taken from patients going to the out-tolerant Department of Obstretric and Gynecology and further shipped off Department of Pathology for cytomorphological examination. The legitimate example assortment is perhaps the main strides in pap smear screening. At any rate one half to 66% of bogus negatives are the aftereffect of patient conditions present at the hour of test assortment and accommodation and the expertise and information on the person who acquires the example. Sufficient cervical cytology tests ought to be gathered and submitted to the lab with proper clinical data. Cross sectional observational study. 27 cases of epithelial cell abnormality, out of which 17 cases show Atypical squamous cells of uncertain significance, 1 case show Atypical glandular cells of uncertain significance. 2 cases shows low grade squamous intraepithelial lesions, 2 cases shows high grade squamous intra epithelial lesions. A total of 5 smears show invasive carcinoma cervix.Maximum cases of LSIL were detected in 41-50 years of age group and maximum cases of HSIL were detected in 31-60 years of age group whereas maximum cases of carcinoma cervix were detected in 51-60 years of age group. In the developing countries like India, cervical cytology, due to its low cost and easy availability, is the most important diagnostic tool for the screening of females of reproductive age group. The patient attending the Gynecology Out Patient Department were targeted in this study, so as to screening all the females, even with minor symptoms like discharge per vaginum.

Association between Gene Promoter Methylation and Cervical Cancer Development: Global Distribution and A Meta-analysis.

DNA methylation is the main epigenetic event for gene silencing and is associated with carcinogenesis. In this meta-analysis, we evaluated the association between the methylation of the promoter regions of APC, CADM1, CCNA1, CDH1, DAPK, FHIT, HIC1, MAL, MGMT, hMLH1, P16, PAX1, RAR-β, and RASSF1 genes and the risk of cervical cancer development and progression. Overall, 194 eligible studies were identified assessing the associations of promoter methylation status of aforementioned genes with low- and high-grade squamous intraepithelial lesions (LSIL and HSIL) and cervical cancer development. The majority of studies were conducted on Caucasian and Asian populations, whereas rare studies were available on the African population. Promoter methylation frequencies were shown to be significantly higher in LSIL and HSIL cervical cancer cases as compared to control specimens for CADM1, CCNA1, CDH1, DAPK1, FHIT, MAL, P16, PAX1, RAR-β, and RASSF1 genes. A moderate association was found between HIC promoter methylation, whereas APC, MGMT, and hMLH1 promoter methylation was not correlated with cervical cancer development. Promoter methylation could be considered as a noninvasive biomarker for early cervical lesions, making them highly promising targets for a personalized therapeutic approach.

A study of papanicolaou stain and cervical acid phosphatase papanicolaou stain in the detection of cervical intra epithelial neoplasm a comparative study

Introduction: In order to increase the sensitivity of the routine PAP test, a new test was introduced called the CAP-PAP test. In this test, lysosomal enzyme and cervical acid phosphatase (CAP) enzyme are labeled with conventional PAP test which will help in detecting abnormal squamous cells of cervix. This enzyme can be used as a biomarker for cervical dysplasia when CAP positivity is detected inside abnormal squamous cells of cervix and can also be used as a surrogate in detection of CIN. Aims: To perform CAP-PAP and PAP staining on cervical smears. To compare CAP-PAP results with PAP results and correlate with cervical histopathology reports. Methods and Materials: 150 females undergoing cervical cancer screening at OBG Department, of our institute were studied. Two cervical smears each were collected from all patients for PAP and CAP-PAP staining. The results were compared and correlated with respective histopathology reports.| Statistical Analysis Used: Chi-square test, Kappa statistics, P value of Results: 75 PAP positive cases out of which 34 were LSIL, 17 were HSIL, 13 were ASCUS, 9 were ASC-H, and 2 were SCC cases. In the remaining 75 PAP negative smears, 38 cases were inflammatory smears and 37 cases were of NLIM smears. Among the PAP positive cases, CAP-PAP results were similar to PAP results in all the LSIL, HSIL, ASCUS, ASC-H and SCC cases. Among the PAP negative cases, CAP-PAP was negative in all the 37 NILM cases, but of the 38 inflammatory cases, CAP-PAP was negative in 30 cases but was positive in 8 cases. CAP-PAP results had good agreement with PAP results and correlated well with histopathology results. Conclusion: CAP positivity is identified by presence of res granules in abnormal squamous cells which helps in its early detection. In screening of cervical cancer on a large scale. CAP PAP acts as a quick and efficient method. The test promises a great future in health centers as the technicians can be easily trained for identification of abnorma

Cytological Evaluation of p16Ink4a in Precancerous Lesions of the Cervix: Conventional Papanicolaou Smears

Background: Cancer of the uterine cervix is the second most common cancer among women worldwide. More than three fourths of these patients are diagnosed at advanced stages, leading to poor prospects of long term survival and cure. Introduction of Papanicolaou (Pap test) cytological screening for cervical precancerous lesions has significantly reduced the incidence and mortality of cervical cancer. Human papillomavirus (HPV) DNA testing has been recently demonstrated to be efficient to be integrated into screening programs, so it can be used to triage women with equivocal cytological abnormalities and also identifies women at risk of residual or recurrent disease after treatment of cervical intraepithelial neoplasia. However, it fails into triage low grade lesions. HPV DNA test confirms infection by the virus, present in 99% of cases. However, it does not discriminate between transient and persistent infection as it is crucial because it is the persistent infection which progresses to neoplasia. Histological assessment of cervical biopsy that is considered as “gold standard” can be hampered by intra and inter observer variability. A more specific triage marker is required to identify women who would need colposcopy. Hence, p16INK4a has emerged as a new diagnostic and prognostic biomarker. Aims and objectives: This study was conducted to evaluate the utility of staining of p16INK4a on conventional Pap smears and its comparison with corresponding biopsies. Material and Methods: 50 cases of conventional Papanicolaou stained cervical smears cases were randomly selected from the archive of cytopathology laboratory. The cervical smears were re-evaluated for adequacy, preservation of cells, cytomorphology and various lesions were categorized according to The Bethesda system 2001 (TBS) classification. Consecutive such cases were selected for which both cytological and histological material were available. Immunostaining of cervical cytological specimens for p16 was performed using monoclonal murine antibody, clone 16P04, JC2. Ready to use antibody was used for immunostaining as per manufacturer's protocol. Results: Out of the 50 smears of preneoplastic and invasive lesions, ASCUS was seen in 5 cases, LSIL in 10 cases, HSIL in 20 cases, SCC in 20 cases and AGUS in 5 cases. Of the 5 cases of ASCUS, histopathology showed 3 chronic cervicitis and 1 each in CIN-1 and CIN-2. Histopathological diagnosis of 10 cases of LSIL showed 6 CIN-1 and 2 each in CIN-2 and chronic cervicitis. Similarly, for 10 cases of HSIL, 3 were CIN-2, 5 CIN-3 and 2 SCC. All 20 cases of carcinoma showed SCC while 5 cases of AGUS showed 4 chronic cervicitis and one adenocarcinoma. Immunohistochemical staining of p16INK4A showed weak positivity in 3 cases of chronic cervicitis and 4 cases of CIN-1. In CIN-2 cases, 66.67% showed strong positivity, CIN-3 showed 80% while both carcinoma and adenocarcinoma showed 100% strong positivity. The sensitivity of immunohistochemical staining of p16INK4A was 77.5%, specificity was 100%, PPV 100% and NPV 52.6%. The relation between histological and cytological immunostaining of p16INK4A. Conclusion: p16INK4A is a reliable marker for dysplastic squamous and glandular cervical cells both in tissue sections and in cervical smears. p16 immunostaining can be easily performed on CPS, and there is high concordance of positivity on smears and tissue sections.

Correlation between HPV-negative cervical lesions and cervical microenvironment

ObjectiveTo investigate the correlation between high-risk human papillomavirus (HR HPV)-negative cervical lesions and cervical microenvironment in Inner Mongolia, China, and to find the pathogenic factors of HR HPV-negative cervical lesions. Materials and methods74 cases of HR HPV-negative healthy women and 80 cases of patients with cervical lesions (28 cases of LSIL, 49 cases of HSIL and 3 cases of CSCC) were selected as the study group; 26 cases of HPV-positive women and 352 cases of patients with cervical lesions (108 cases of LSIL, 214 cases of HSIL and 30 cases of CSCC) were control group. Questionnaires were collected from the study group and the control group and specimens were collected. Gram staining, hematoxylin and eosin staining microscopy, and substrate colorimetry method were used to detect vaginal micro-ecological indicators; ELISA was used to detect the concentration of SIgA, IgG, IL-2 and IL-10 in vaginal lavage fluid. Genetic testing was used to detect HPV, mycoplasma, and chlamydia infection. The changes of vaginal micro-ecology evaluation index and local immune factor concentration in healthy women and cervical lesions of all grades in the study group and the control group were compared. ResultsPatients with cervical lesions, compared with healthy women, had a decrease in dominant lactobacilli and dysbacteriosis (P < 0.05), and this trend became more apparent as the disease progressed. The diversity and concentration of the flora in the HPV-negative group increased, the abnormal composition ratio decreased, and the HPV-positive group showed the opposite trend. As the lesion progressed, H2O2 decreased first and then increased, and the overall trend of SNa, LE, GUS, and GADP increased. The infection rate of trichomoniasis, BV and chlamydia increased and infection rate of Candida decreased. Also, compared with healthy women, patients with cervical lesions showed changes in immune factor concentration (P < 0.05). As the lesion progressed, IL-2 decreased, IL-10 increased, and IL-2/IL-10 decreased. However, IL-2 expression in HPV-negative group was higher than HSIL. SIgA was significantly lower in patients with cervical lesions than in healthy women. IgG had an upward trend in the HPV positive group. ConclusionThis study showed that vaginal micro-ecological imbalance and weakening of local cervical immune function are important reasons for the development of cervical lesions. It is expected to inhibit the development of cervical lesions by regulating the balance of vaginal micro-ecology and enhancing local immune function. By detecting Lactobacillus vaginalis, pre-enzyme, IL-2, IL-10, SIgA, it can guide the further diversion of HPV-positive women and predict the development direction of cervical lesions after HPV infection.

Correlation of High-Risk HPV Genotypes with Pap Test Findings: A Retrospective Study in Eastern Province, Saudi Arabia

Introduction: High-risk human papillomavirus (HR HPV) is found to be responsible for 4.5% of cancer in general, primarily cervical cancer. We aim here to highlight the prevalence and genotypes of HR HPV and correlate its association with Pap tests’ results, which are still not well known in the Eastern Province of Saudi Arabia. Methods: Over 7 years (2013–2019), the results of 164 Saudi women coinvestigated for HR HPV along with Pap tests were collected from the archive of King Fahd University Hospital. Only women who had atypical squamous cells of undetermined significance (ASCUS) on the Pap test and those at elevated risk of infection were cotested for HR HPV; otherwise, the Pap test was the only screening modality for cervical cancer. Data were organized and statistically analyzed using IBM SPSS v26. Results: Out of 164 Saudi women, 14.5% (n = 24/164) showed positive results for HR HPV (8 patients had HPV16 and 2 had both HPV16 and HPV18/45, while the remaining 14 had other HR HPV); among them, 41.5% (n = 10/24) had an abnormal Pap test (5 ASCUS and 5 LSIL), while 58.5% (n = 14/24) had a negative Pap test. On the other hand, 21% (n = 35/164) of patients, in the study, had an abnormal Pap test (24 ASCUS, 8 low-grade squamous intraepithelial lesion [LSIL], and 3 atypical glandular cell [AGC]). In 80% (n = 19/24) of ASCUS cases, HR HPV was not detected; however, 20% (n = 5/24) were positive for other HR HPV. Concerning LSIL cases, 62.5% (n = 5/8) were positive for HR HPV (1 case showed HPV16 and HPV18/45, 2 cases showed HPV16, and 2 cases showed other HR HPV), while in the remaining 37.5% (n = 3/8) LSIL cases, HR HPV was negative; similarly, all AGC cases were negative for HR HPV. Statistical analysis showed a significant correlation between HPV status and Pap test findings (p value <0.001). Discussion/Conclusion: HR HPV frequency and genotype distribution, in this study, might reflect a different regional infection pattern. The high association of HR HPV with negative cytology emphasizes the need to add the HR HPV test to screening modalities of cervix cancer.

Assessment of micronuclei counts as tumour marker in cervical carcinogenesis: A follow-up study.

Micronuclei counts were performed in cervical smears with low-grade squamous intraepithelial lesions of the cervix (LSIL) to assess its potentiality as tumour marker in cervical carcinogenesis. The cases studied were from the ongoing rural cervical cancer screening in west Lucknow, India. Micronuclei counts were performed in the cervical smears of 100 LSIL cases, and the number of cells with micronuclei was defined as micronucleated cells (MNC) and the number of micronuclei per 1000 cells as MNC score. Human papillomavirus (HPV) DNA testing was also done in 100 LSIL cases by GeneNav qPCR test. A high MNC score was found in 20 of the 100 LSIL cases while the counts were low in the remaining 80. Persistence of LSIL was seen in 19 of the 20 LSIL cases with high MNC score while only six cases of the 80 cases with low MNC score showed persistence. The persistence of LSIL was very high in cases with high MNC score. The multiple high-risk HPV types such as 18, 31, 33 and 35 were seen in 12 of the 100 LSIL cases and a high positivity rate was seen in women with high MNC score. The persistence of LSIL was also higher with HPV positivity. The study revealed correlation between high MNC score, persistence of LSIL and HPV positivity. Hence, MNC score can prove to be very useful in discriminating high-risk LSIL cases that are less likely to regress and possibly may progress to high-grade squamous intraepithelial lesions or carcinoma.

Cytomorphological spectrum of cervical smears in a tertiary hospital

Cervical cancer is one of the leading causes of mortality and morbidity in women worldwide. Invasive cervical carcinomas are preceded by a stage in which the abnormal cells are confined to the epithelium (Intraepithelial stage). Women can be routinely screened for intraepithelial stage with cervical cytology smears and hence invasive stage can be prevented by early diagnosis. Cervical cytology smears also an useful tool in aiding the diagnosis of infectious and inflammatory conditions of the cervix.To evaluate the cytomorphological spectrum of cervical smears referred to a tertiary hospital.It is a retrospective observational study. Clinical data and pap smear cytology reports were obtained from the archives during the period of May 2018-May 2019(1 year study). All smears were reported as per “Bethesda system of reporting cervical cytology ” 2001.A total of 1241 cases were examined. The age group of patients ranged between 18yrs to 75 yrs. A total of 1203 cases (97%) were reported as Negative for intraepithelial lesions/malignancy, out of which 124 cases(0.1%) showed vaginal candidiasis, 56 cases (0.05%) were reported as Trichomonas vaginalis infection, 321 cases (0.26%) as Bacterial vaginosis, 1 case of HSV infection associated changes, 67 cases(0.05%) were reported as inflammatory smears. 38 cases showed abnormality, out of which 29 cases were of ASC-US,3 cases of LSIL, 3 cases were reported as HSIL, 2 cases as Squamous cell carcinoma and 1 case with adenocarcinoma cervix.Cervical inflammatory lesions (including infections) and neoplastic lesions (includes intraepithelial and epithelial malignancies) can be diagnosed by Cervical cytological smears.