Cervical Biopsy Research Articles

Detection of high-risk human papillomavirus infected cervical biopsies samples by immunohistochemical expression of the p16 tumor marker.

Cervical cancer is the fourth most common type of cancer in women worldwide. It is widely accepted that the main cause of cervical cancer, especially in underdeveloped countries like Pakistan, is the infection caused by the human papillomavirus (HPV). The current screening and diagnostic methods face several challenges in accurately detecting the various types of lesions caused by HPV. Therefore, the present study was conducted to assess the effectiveness of p16 immunohistochemistry (IHC) analysis as a diagnostic method in samples of cervical biopsies. One hundred cervical biopsy samples were obtained from female patients across various age groups (> 20- ≤ 30, > 31- ≤ 40, > 41- ≤ 50, > 51- ≤ 60years). These samples were subsequently prepared for subsequent examination. All samples were analyzed using automated tissue processing followed by Hematoxylin and Eosin (H & E) staining, and p16 IHC tumour marker staining. The H & E slides showed changes in normal cervical tissues, while four cervical abnormalities were identified statistically significant using p16 marker including chronic cervicitis, nabothian cyst formation, cervical intraepithelial neoplasia, and cervical cancers (P value 0.014). Furthermore, among females of different age groups (> 31- ≤ 40, > 41- ≤ 50, > 51- ≤ 60years) were found statistically significant suffering from cervical cancer (P value 0.04), HPV with cervical cancer (P value 0.01), HPV with cervical intraepithelial neoplasia (P value 0.01). Based on the available data, it can be inferred that the incorporation of the p16 tumor marker may be a valuable method for detecting high-risk HPV in cervical biopsies samples.

Upregulation of IFNE in cervical biopsies of patients with high-risk human papillomavirus infections.

Interferon epsilon (IFN-ε) is constitutively expressed in the epithelium of female reproductive tract and confers vital protection against sexually transmitted pathogens in mouse models. However, there is limited insight into the role of IFN-ε in human sexually transmitted infections such as human papillomavirus (HPV). Cervical biopsies were obtained from high-risk (HR) HPV positive (n = 28) and HR-HPV negative (n = 10) women. mRNA expression of IFN-ε in cervical tissues was measured by qPCR. Expression of the IFN-ε protein was determined by Western blot analysis, immunohistochemistry and immunofluorescence staining. mRNA expression of IFN-ε was higher in the ectocervix than that of other IFNs, and was further upregulated in HR-HPV positive women compared with HR-HPV negative women. Expression of the IFN-ε protein was comparable between HR-HPV infected patients and healthy controls. These results reveal differential expression of IFN-ε mRNA between individuals with or without HR-HPV infection, and imply direct or indirect regulatory mechanisms for IFN-ε transcription by HPV. Expression of IFN-ε protein in HPV infections would require further validation.

HPV18 L1 and long control region sequences variation and E6/E7 differential expression in nasopharyngeal and cervical cancers: a comparative study

BackgroundThe role of high-risk human papillomaviruses (hr-HPVs) in cervical cancer (CC) pathogenesis has long been established. Knowledge about the involvement of hr-HPVs in the etiology of nasopharyngeal cancers (NPC) was not well appreciated until the early 2000s when a clear link began to emerge. However, it is not clear whether HPV oncogenesis in the different epithelial cancers is associated with L1 gene and long-control region (LCR) sequences variation. This study aimed to investigate the HPV18 L1 gene and LCR sequences variation in cervical and nasopharyngeal biopsies, and assessed E6 and E7 genes expression level in both cancers.MethodFour-hundred and three (403) formalin-fixed paraffin-embedded tissues originating from nasopharyngeal (NPC) (279) and cervical (CC) (124) sites were collected from a pathology laboratory, Pathologist Without Borders, Accra, Ghana. Haematoxylin and eosin staining was carried out to confirm the presence of cancer on prepared biopsy sections. DNA was extracted from the confirmed cancer biopsies, followed by PCR using MY09/GP5+ /6+ primers to detect the presence of HPV and specific primers for the amplification of L1 gene and LCR. Sanger sequencing was carried out to determine HPV genotypes, and L1 and LCR sequences variant of HPV18s in CC and NPC biopsies. The HPV18 E6/E7 mRNA expression pattern in both cancers was determined using RT-qPCR.ResultsMost of the NPC (45%) and CC (55%) biopsies were HPV18 positive. Comparison of HPV18 L1 sequences obtained from cervical and nasopharyngeal cancer tissues, the L1 sequences from the NPC were highly dissimilar with a 59–100% variation among themselves, and in relation to the reference strains. However, the L1 sequences from the CC were more similar with a 91.0–100% variation among the amplified sequences. Also, the LCR sequences from CC were quite different relative to that of NPC. Results for the differential expression of E6/E7 in the two cancers showed a higher fold change in E6 expression in the CC tissues than the NPC tissues while a reverse expression pattern was found for E7 gene.ConclusionThe current study reports for the first-time variations in HPV18 L1 and LCR sequences, and differential expression of E6/E7 genes in NPC compared to CC, suggesting a possible adaptation mechanism of the virus at different cancer sites.

Is Forced Coughing Effective in Reducing Pain During Cervical Biopsy?

This review aimed to compare the potential analgesic effect of forced coughing (FC) with that of local anaesthetics (LA) or placebo during cervical biopsy. A total of 5 electronic databases—Scopus, PubMed, Web of Science, Cochrane Library and Google Scholar—were systematically searched from inception till March 2021. Data were extracted from 6 randomised controlled trials and analysed. During cervical biopsy, the overall effect favoured LA over FC (mean difference [MD] = 1.06, 95% confidence interval [CI]: 0.58 to 1.54; P <0.0001). Compared to no pain management, pooled data were comparable between the two groups (MD = −1.2, 95% CI: −3.35 to 0.94; P = 0.27). Procedure duration was significantly longer in the LA group than in the FC group (MD = −1.94, 95% CI: −2.47 to −1.41; P <0.00001). FC and LA are both useful pain-lowering modalities during cervical biopsy, depending on the setting and their availability. Keywords: Biopsy; Colposcopy; Anesthesia; Pain.

A rare case of cervical gastric-type adenocarcinoma mimicking bilateral ovarian malignancy in a genetically confirmed Peutz-Jeghers syndrome without known polyposis

Abstract Introduction/Objective Cervical gastric-type adenocarcinoma (GAS) is rare and accounts for 2-4% of cervical cancer. GAS is usually sporadic but can be associated with Peutz-Jeghers syndrome (PJS), a rare autosomal dominant disorder caused by germline STK11 mutations and characterized by hamartomatous polyposis in the gastrointestinal (GI) tract, mucocutaneous pigmentation, and cancer predisposition. Although cases of GAS may occur in patients with PJS associated with polyposis, our patient with GAS has an unusual and perhaps unique association with the PJS mutation that is unassociated with polyposis. Methods/Case Report A 33-year-old female presented with abdominal pain, abnormal uterine bleeding, vaginal discharge, weight loss, and fatigue for one year. Pigmentations were noted on lips, gum, tongue, and hands. She had a family history of maternal ovarian and pancreatic cancers. Computerized tomography revealed large bilateral multiloculated complex cystic adnexal masses, thickened endometrium extending into the endocervix, and ascites. Tumor markers showed increased CA19-9 and CA125, suggesting either a primary ovarian malignancy or possible metastatic GI tract cancer to both ovaries. Her PAP smear demonstrated atypical glandular cells that favored endocervical origin. HPV was not detected. A cervical biopsy showed bland gastric-type glands without obvious stromal invasion. Given the clinical concern for malignancy, a cervical cone biopsy was performed, demonstrating unequivocal invasion by gastric-appearing glands. The tumor cells were positive for CK7, CEA, and PAX8, and were negative for CK20, CDX2, and p16. The pathologic diagnosis confirmed a primary GAS with a spread to bilateral ovaries. Although the upper GI tract and left colon were free of polyps or masses by endoscopic exam, the genetic evaluation revealed a germline heterozygous STK11 mutation consistent with PJS. Results (if a Case Study enter NA) NA Conclusion To our knowledge, this is the first case of occult PJS (confirmed by genetic analysis) presenting with widely advanced metastatic GAS without known polyposis.

Understanding cytomorphologic changes in Pap tests of transgender men on testosterone therapy

Abstract Introduction/Objective Pap test findings in transgender men on testosterone therapy can often lead to misdiagnosis of dysplasia, especially in the setting of a positive HPV test. We reviewed relevant literature to analyze the cytomorphological nuances and applicable correlations to subsequent pathology in Pap tests of transgender men on testosterone therapy. Methods/Case Report We searched peer-reviewed journals on PubMed, synthesizing five articles published from 2018-2022 that specifically address cytomorphological findings on cervical Pap tests and biopsy results in transgender men on testosterone therapy. Results (if a Case Study enter NA) Various cytological changes, such as small parabasal-type cells and transitional cell metaplasia (TCM) can be mistaken for dysplasia in transgender men. In fact, these changes are representative of atrophy due to testosterone, as such findings have been observed in patients undergoing tamoxifen therapy. Small parabasal-type cells are described as clusters or sheets with intermittent nuclear enlargement and hyperchromatic nuclei, whereas TCM comprises flat sheets of cells with oval to spindle-shaped nuclei, powdery chromatin, and prominent nuclear grooves. Relevant clinical history can often help clarify cytologic diagnoses, as Pap tests that show atrophic changes can be misinterpreted as being abnormal. In several cases described in the literature, original diagnoses of atypical squamous cells of undetermined significance (ASCUS) were re-interpreted as atrophic changes after eliciting a history relevant for testosterone therapy. Small parabasal-type cells found on cytology have correlated with sections of the ectocervix that have groups of small hyperchromatic cells attached to the surface of atrophic cervical epithelium or areas of TCM. These cells may be mistaken for endometrial cells, especially in patients over 45 years old. Distinguishing atrophic changes from dysplasia on surgical pathology may modify cytologic diagnoses. Conclusion Transgender men on testosterone therapy often retain their cervix, necessitating ongoing cervical cancer screening. However, Pap test results from these individuals often exhibit atrophic changes through the presence of small parabasal-type cells and TCM. Differentiating between atrophy and dysplasia can pose a diagnostic challenge, especially in the setting of positive HPV testing or inflammation. Continued analyses into these cytomorphological nuances will assist providers in refining their diagnostic approaches and providing personalized care to transgender men.

The relationship between the expression of Stathmin and tumor immune cell infiltration in primary cervical carcinoma and its prognostic diagnostic value

To analyse the relationship between the Stathmin expression and the tumor immune cell infiltration in primary cervical carcinoma (CC), and its prognostic diagnostic value. Cervical tissue samples from 128 patients admitted to our hospital from February 2021 to February 2022 who underwent hysterectomy or cervical biopsy were selected as the observation objects, and then divided into control group (normal cervical tissue specimen, n = 30), cervical intraepithelial neoplasia (CIN) group (CIN neoplasia cervical tissue specimen, n = 30), and cervical cancer group (cervical tissue specimen of cervical cancer lesion, n = 68) according to pathological results. The expression of Stathmin was detected by Immunohistochemistry (IHC). The numbers of infiltrated neutrophils (TINs) were measured by the specific esterase staining. The pathological data of CC patients were collected, and the protein expression of Stathmin was compared with different pathological data. According to the protein expression of Stathmin, the samples were divided into Stathmin positive group and negative group. The infiltration numbers, the levels of CD3+T, CD4+T and CD8+T of TINs were compared in each group, and CD4+/CD8+ were calculated. Kaplan-Meier survival curve was used for the analysis of the relationship between the Stathmin expression and the clinical prognosis in CC. COX analysis was used to determine the factors affecting the prognosis of patients with CC. The proportion of positive expression of cervical tissue Stathmin in CIN group and cervical cancer group was significantly higher than that in the control group, and the proportion of positive expression of cervical tissue Stathmin in cervical cancer group was significantly higher than that in CIN group (P < 0.001). The ratio of the clinical stage II + III, tissue grade low and medium differentiation, and lymph node metastasis in Stathmin positive group was significantly higher than that in the Stathmin negative group (P < 0.05). Compared with the Stathmin negative group, the numbers of TINs infiltration and the level of CD4+/CD8+ were visibly higher, and the content of CD3+T, CD4+T and CD8+T were sharply lower in the Stathmin positive group (P < 0.001). Compared with the Stathmin negative group, the numbers of TINs infiltration were higher, but the content of CD4+T and CD8+T were lower in Stathmin-positive group (P < 0.001). The 68 CC patients were followed up, with a median follow-up time of 25 months (5–60 months) and an overall survival rate of 66.18 % (45/68). The results of Kaplan-Meier survival curve showed that the median survival time of patients with the Stathmin positive expression was 30 months, which was significantly lower than that of 46 months (P < 0.05) in patients with the Stathmin negative expression. Multivariate analysis of COX proportional regression risk model showed that the Stathmin positive expression, TINs infiltration numbers≥55, CD3+<5.5 %, CD4+<4.5 %, CD8+<3 %, CD4+/CD8+≥1.3 were independent factors affecting the prognosis of CC patients (P < 0.05). Stathmin in the primary tissue of CC had a significantly high expression state, which was involved in the occurrence and development of CC, thus affecting the immune microenvironment balance damaged by tumor immune cell infiltration. Detecting its expression level might help the diagnosis and prognosis assessment of CC, and Stathmin might become a new target of CC immunotherapy.

The Mulher Study: cervical cancer screening with primary HPV testing in Mozambique

ObjectiveTo evaluate cervical cancer screening with primary human papillomavirus (HPV) testing in Mozambique, a country with one of the highest burdens of cervical cancer globally.MethodsWomen aged 30–49 years were prospectively...

Associations of multi-human papillomavirus infections with expression of p16 in a cohort of women who underwent colposcopy: a retrospective study of 5165 patients.

Investigate HPV types in cervical specimens, their correlation with p16 expression in lesions, and diagnostic value for cervical lesions. Enhance clinical diagnosis reliability. Retrospective cross-sectional study at Fujian Maternity and Child Health Hospital's Cervical Disease Center (Jun 2019-Dec 2021). Patients with abnormal cervical screening underwent colposcopy and conization. Pathological diagnosis based on colposcopy, cervical biopsy, ECC, and conization. Analyzed HPV genotyping (18 HR-HPV, 5 LR-HPV) and p16 expression correlation. Statistical analysis used R software. he expression of p16 is significantly associated with the infection of high-risk HPV types, such as 16, 33, 52, and 58, with an increased risk of 1.4 times or higher (OR=1.91, 3.14, 1.40, and 1.78, respectively). The risk of p16 expression increased 4-fold for multiple high-risk HPV types [adjusted OR (95% CI) = 4 (2.92~5.5), P-value <0.001]. Compared to the p16(-) group, the p16(+) group had a higher association with cervical lesions worse than HSIL (High-grade Squamous Intraepithelial Lesions).In the group with multiple Human Papillomavirus Infections with types 16, 33, 52, and 58, the risk of cervical lesions worse than HSIL increased by up to 660-fold compared to the negative group (adjusted OR=660.62, 95% CI: 91.39~4775.53, P<0.001), indicating that this combination of HPV types posed the greatest risk for cervical lesions above HSIL. p16 plays a crucial role in cervical lesion progression, linked to high-risk HPV. Combining p16 with HPV screening improves cervical cancer detection. Studying multiple HPV infections will enhance prevention and management.

Assessing a relation between composition of luminal and tissue microbiota and cervical intraepithelial changes

The incidence of cervical cancer (CC) in Russia remains at a high level and ranks fourth among all cancers. CC is preceded by mild, moderate, and severe cervical intraepithelial neoplasia. Human papillomavirus (HPV) is known to be the main cause for its development being responsible for 99% cancer cases. Despite HPV infection, the oncological process occurs only under certain conditions. There are risk factors that indirectly affect the course and emergence of cervical dysplasia. In recent years, the role of cervico-vaginal microbiome for onset and progression of this pathology has been actively discussed. Many studies evidence that dysbiosis along with cervical intraepithelial dysplasia is associated with increased HPV viral load, additionally allowing to clarify a role of specific microorganisms. Despite the knowledge rapidly accumulating about the nature of vaginal microbiome in cervical precancerous processes, the level of its impact on disease course has not been fully investigated and is of great interest. Identification of microorganisms that affect emergence and progression of this pathology will allow to prevent and apply select approach to treatment of dysbiosis. The aim of our study was to identify a relation between cervical tissue and luminal culturome and severe cervical intraepithelial neoplasia. The study evaluated the microbiota of cervical and cervical canal mucosa biopsy in patients with cervical intraepithelial changes. According to the study results, the Streptococcus genus representatives were significantly more frequent in cervical biopsy specimens in severe dysplasia vs. no cervical intraepithelial changes, whereas cervical canal cultures provided comparable data. Bacteria from the Corynebacterium genus were found in cervical biopsy specimens 3 times more often than in smear from the cervical canal mucous membrane. The microbiota from cervical and cervical canal biopsy specimens differ qualitatively and quantitatively particularly regarding prevalence of Streptococcus spp. representatives being more common in patients with cervical dysplasia. Corynebacterium spp. were detected more often in cervical biopsy specimens than in cervical canal smears. Thus, the method of collecting biological material has a great influence on final results.

Study of the role of various diagnostic methods in patients with benign and neoplastic cervical diseases

Introduction. Cervical intraepithelial neoplasias that have been recognized as a precursor of squamous cell carcinoma are subject to active detection for the purpose of eradication. Human papillomavirus (HPV) exposure of women is the major risk factor for carcinogenesis, which is a necessary part for the formation of a pre-tumour and malignant phenotype. Cervical intraepithelial neoplasias that precede the development of squamous cell carcinoma should be actively detected and treated early. The identification of patients with HPV is an important part of subsequent therapies. Inosine pranobex is used as one of the components of the treatment of HPV-associated diseases.Aim. To study the role of various diagnostic methods in patients with benign and neoplastic cervical processes.Materials and methods. The study included 311 women who were 21–64 years old. Colposcopy, cytological examination, HPV testing, preliminary biopsy, cervical excise with histological evaluation of tissue sampling were carried out.Results. HPV 16 was more often detected in a malignant tumour and CIN 3; HPV type 31 and 33 were more often detected in CIN 2 and CIN 3; HPV type 18 was detected in almost all patient with CIN, and malignant and benign neoplastic disorders in preliminary biopsy, that was not typical of results of the excision. In excision CIN 1 were confirmed in 17.07%, CIN 2 in 31.88%, CIN 3 in 66.67%. Preliminary results of benign process in 6.67% turned out to be CIN 1, and in 6.67% – CIN 2, that confirms the need for a comprehensive evaluation. Studies of prevalence of various types of HPV showed that the established sequence of recommended tests, which consists of extended colposcopy, cytology and identification of HPV types, if necessary, does not always correspond to the biopsy results and histological findings on cervical excision specimens. According to our sources, the final result can be determined after excision procedure of cervical tissue and histological examination of the tissue obtained.Conclusion. No strong correlations between examination methods and cervical pathology have been revealed. The final diagnosis is made after cervical biopsy. High prevalence of different types of HPV raises the question of need for complex therapy with inclusion the inosine pranobex.

Nurses and doctors ‘s awareness and knowledge towards treatment and care of cervical cancer patients: a cross-sectional study

AimThe present study aimed to investigate healthcare professionals’ perceptions and experiences in caring for cervical cancer patients. The present study was also designed to assess the healthcare professionals’ attitudes toward cervical cancer screening and its prevention.MethodsA cross-sectional quantitative descriptive study was conducted, and 540 participants (240 nurses and 300 doctors), from different hospitals of Pakistan have been selected and interviewed.ResultsData was collected using structured questionnaires and SPSS was used to statistically analyze the data. Participants in the present study are questioned with respect to age, gender, and work experience. The mean age of the participants is 35 years. Among them, 41% of participants are < 35 years of age and 59% are > 35 years of age. In the case of gender, 22% of participants are males and 78% are females. 47% of the participants have work experience < 20 years and 53% have work experience > 20 years. Data from the present study showed that most of the nurses are less educated (basic education of middle and matric degree) with a simple diploma in nursing and midwifery. Nurses and doctors do not have any knowledge/experience of the patient’s psychological counselling. Participants are also questioned with respect to HPV vaccination, 39% of nurses and 62% of doctors are vaccinated. The difference in vaccination frequency of participants was observed as statistically significant (p < 0.0001). In the case of treatment modalities, doctors have statistically more knowledge about the pap smear (p < 0.0001), cervical biopsy (p < 0.0001), colposcopy (p < 0.0001), and visual application after acetic acid application (p < 0.0001) compared to nurses. Data analysis showed that Pap smear was performed significantly higher in married females compared to unmarried (p < 0.0001).Conclusionour study provides a comprehensive and in-depth perspective of the nurses and doctors for cervical cancer patients. Cervical cancer prevalence is increasing due to inadequate knowledge and awareness among healthcare professionals. Improvement can be brought about by the regular use of treatment modalities in unmarried females also.

К ВОПРОСУ ОБ ОЦЕНКЕ КУЛЬТУРОМА БИОПТАТА ШЕЙКИ МАТКИ У ПАЦИЕНТКИ С ЦЕРВИКАЛЬНОЙ ИНТРАЭПИТЕЛИАЛЬНОЙ ДИСПЛАЗИЕЙ УМЕРЕННОЙ СТЕПЕНИ

In recent years, the role of microcenosis in the occurrence and progression of the pathology of various organs, including cervical cancer, has been actively discussed. Purpose: to study the cervical microflo-ra in a patient with moderate cervical dysplasia. Material and research methods. An extended micro-biological study of cervical mucosal cultures and cervical biopsy was performed in a patient with moderate cervical dysplasia. Cervical biopsy microcenosis was studied using a MALDI-ToF mass spectrometer (Bruker). The biocenosis of the cervical canal was studied on the basis of the polymerase chain reaction method, the Digene HPV test system test and bacteriological examination. Research results. In the biopsy specimen of the cervix, a microflora not typical for the cervical biotope was found. In particular, microorganisms were found in the tissue of the cervix, which can cause the de-velopment of pathology of the oral cavity (Streptococcus oralis, Streptococcus mitis, Streptococcus anginosus, Streptococcus sanguinis) or intestinal dysbiosis (Alloscardovia omnicolens). At the same time, these bacteria were not found in the mucous membrane of the cervical canal. Conclusion. Fur-ther research is needed to understand the causes and consequences of biocenosis differences in the altered tissue of the cervix and cervical canal.

P16 expression and recurrent cervical intraepithelial neoplasia after cryotherapy among women living with HIV.

The expression of p16 protein, a surrogate marker for high-risk human papillomavirus (hrHPV), is associated with cervical dysplasia. We evaluated correlates of p16 expression at treatment for high-grade cervical lesions and its utility in predicting the recurrence of cervical intraepithelial lesions grade 2 or higher (CIN2+) following cryotherapy among women with HIV. This is a subgroup analysis of women with HIV in Kenya with baseline cervical biopsy-confirmed CIN2+ who were randomized to receive cryotherapy and followed every six-months for two-years for biopsy-confirmed recurrence of CIN2+. P16 immunohistochemistry was performed on the baseline cervical biopsy with a positive result defined as strong abnormal nuclear expression in a continuous block segment of cells (at least 10-20 cells). Among the 200 women with CIN2+ randomized to cryotherapy, 160 (80%) had a baseline cervical biopsy specimen available, of whom 94 (59%) were p16-positive. p16 expression at baseline was associated with presence of any one of 14 hrHPV genotypes [Odds Ratio (OR) = 3.2; 95% Confidence Interval (CI), 1.03-9.78], multiple lifetime sexual partners (OR = 1.6; 95% CI, 1.03-2.54) and detectable plasma HIV viral load (>1,000 copies/mL; OR = 1.43; 95% CI, 1.01-2.03). Longer antiretroviral therapy duration (≥2 years) at baseline had lower odds of p16 expression (OR = 0.46; 95% CI, 0.24-0.87) than <2 years of antiretroviral therapy. Fifty-one women had CIN2+ recurrence over 2-years, of whom 33 (65%) were p16-positive at baseline. p16 was not associated with CIN2+ recurrence (Hazard Ratio = 1.35; 95% CI, 0.76-2.40). In this population of women with HIV and CIN2+, 41% of lesions were p16 negative and baseline p16 expression did not predict recurrence of cervical neoplasia during two-year follow up.

Comparing the performance of 2 human papillomavirus assays for a new use indication: a real-world evidence-based evaluation in the United States

The US Food and Drug Administration supports innovations to facilitate new indications for high-risk human papillomavirus testing. This report describes the retrospective testing of stored specimens and analysis of existing data to efficiently and cost-effectively support a new indication for the Onclarity human papillomavirus assay (Becton, Dickinson and Company, BD Life Sciences - Integrated Diagnostic Solutions, Sparks, MD). The performance of this index test was compared with that of a predicate test, the cobas human papillomavirus assay (Roche Diagnostics, Indianapolis, IN). Both human papillomavirus assays are based on real-time polymerase chain reaction platforms that detect the presence of 14 high-risk human papillomavirus genotypes. The predicate assay reports human papillomavirus types 16 and 18 as individual results and the other 12 human papillomavirus genotypes as 1 pooled result. The index assay reports 9 independent results (human papillomavirus types 16, 18, 31, 33/58, 35/39/68, 45, 51, 52, and 56/59/66). Both the index and predicate assays are approved by the Food and Drug Administration for cervical cancer screening, but at the time that this study was initiated, the index human papillomavirus assay was not approved for use with cervical specimens collected in PreservCyt (Hologic, Inc, San Diego, CA) liquid-based cytology media. The performance of the index human papillomavirus assay was compared with that of the predicate human papillomavirus assay for the detection of cervical intraepithelial neoplasia grades 2 or greater and 3 or greater (≥CIN2 or ≥CIN3) using PreservCyt liquid-based cytology specimens collected from women aged 21 to 65 years. In addition, the ability of the index test's extended genotyping to stratify ≥CIN2 and ≥CIN3 risks, using these specimens, was evaluated. The New Mexico HPV Pap Registry was used to select an age- and cytology-stratified random sample of 19,879 women undergoing opportunistic cervical screening and follow-up in routine clinical practice across New Mexico. A subset (n = 4820) of PreservCyt specimens was selected from 19,879 women for paired testing by the index and predicate human papillomavirus assays within age and cytology strata and included women with or without cervical biopsy follow-up. Point estimate differences and ratios were calculated for cervical disease detection and positivity rates, respectively, with 95% confidence intervals to determine statistical significance. The cumulative risk of ≥CIN2 or ≥CIN3, with up to 5-year follow-up, was estimated for the index assay using Kaplan-Meier methods. The 5-year cumulative ≥CIN3 detection rates were 5.6% for the index assay and 4.6% for the predicate assay (difference, 1.0%; 95% confidence interval, 0.5%-1.5%). The ≥CIN3 positivity rates within <1 year were 95.3% for the index assay and 94.5% for the predicate assay (ratio, 1.01; 95% confidence interval, 0.98-1.06). The ≥CIN3 cumulative positivity rates for the index and predicate assays were also similar at 5 years. Among cases of ≥CIN3, the positive agreement rates between the index and predicate assays for human papillomavirus types 16 and 18 were 100.0% (95% confidence interval, 95.0%-100.0%) and 90.9% (95% confidence interval, 62.3%-98.4%), respectively. Human papillomavirus type 16 carried the highest ≥CIN2 or ≥CIN3 risk, followed by human papillomavirus types 18/31/33/58/52/45 and human papillomavirus types 35/56/59/51/56/59/66. The index and predicate human papillomavirus assays demonstrated equivalent performance, and extended human papillomavirus genotyping, using the index assay, provided effective ≥CIN2 and ≥CIN3 risk stratification, supporting a new indication for use of the index assay with PreservCyt.

Development of a Machine Learning Model to Evaluate Changes during Radiotherapy in Cervical Cancer Tumor Cells.

Radiotherapy is considered the standard treatment for advanced cervical cancer. It is known that irradiation changes the morphology of tumor cells during the treatment, though its clinical significance is unknown. Recently, the usefulness of machine learning using medical images has been reported, as also as pathological images. The purpose of this study is to create a classification model for cervical tumor biopsy treated with radiotherapy, visualized on whole slide images using machine learning, in an attempt to explore the clinical significance of morphology changes during radiotherapy. Cervical cancer patients who underwent radical irradiation with both pre- and mid-treatment biopsy from April 1, 2013, to December 1, 2020, were retrospectively analyzed. Tumor biopsies were Hematoxylin and eosin stained and digitized to whole slide images (WSIs). WSI s were divided into tiles of 224 × 224 pixels and converted into feature vectors using a pre-trained convolutional neural network (densenet121). A training and test dataset were divided 1:1 on a patient basis, and the model was trained to classify "pre-treatment" and "mid-treatment", and its accuracy was evaluated. The probability obtained from the classification model was defined as the radiation change index (RCI), and its color map was projected on the WSI for visualization. Survival analysis was performed to examine the clinical significance of the RCI values. A total of eighty-four patients were analyzed, and the median observation period was 3.2 years. 184 WSIs were obtained, and 2203407 tiles were generated. The classification model was trained using 2500 tiles for each "pre-treatment" and "mid-treatment". The accuracy of the classification model was 70.8 %, with an AUC of 0.77 for the ROC curve. The probability obtained from the classification model was defined as RCI, with 0 being "pre-treatment" and 1 being "mid-treatment", and the mean RCI of "pre-treatment" and "mid-treatment" were 0.38 and 0.61, respectively. When the RCIs were visualized with a color map, the areas considered to be "pre-treatment" were consistent with a viable tumor component, and the areas considered to be "mid-treatment" were consistent with fibrosis supposed to cause by irradiation. To evaluate the clinical significance of RCI, we divide the clinical cohort into two groups with a threshold mean RCI of 0.4 in the pre-treatment biopsy. The disease-free survival was 91 months and 47 months in the RCI<0.4 and RCI≥0.4 groups, respectively. We created a model to classify tissues before and during radiotherapy and successfully visualized it on slide images. The low RCI group before treatment had a better prognosis than the high RCI group, suggesting that the tumor morphologic features obtained by machine learning may be useful for prognosis prediction. Further studies are planned to develop a model that can accurately predict prognosis.

The impact of intracervical hyaluronidase enzyme on cervical histomorphology in successfully detorted uterine torsion affected buffaloes

The present study evaluated the impact of intracervical hyaluronidase enzyme on cervical histomorphology in successfully detorted uterine torsion affected buffaloes. These animals were either subjected to routine post-detorsion treatment (n=10) or in addition to routine treatment, hyaluronidase enzyme (10,000 IU; 2.5 ml at each of 3, 6, 9, 12 o’clock position of cervix) was administered intracervically immediately post-detorsion (n=10), followed by repeated administration at 6 h interval (0, 6, 12, 18 h) till complete cervical dilatation or till 24 h after detorsion. The pre-treatment cervical biopsy samples were collected immediately after detorsion and post-treatment samples were collected at complete cervical dilatation which happened within 24 h after detorsion in all the cases. Following histomorphology, the quantification of collagen fibers as mean percentage area occupied collagen out of total tissue area in pre-treatment cervical biopsy samples of all the buffaloes was revealed as 21.0±5.7. During post-treatment period, at the time of complete cervical dilatation, this value exhibited a decrease in control (9.1±2.2; P&gt;0.05) as well as hyaluronidase group (5.8±2.1; P&lt;0.05). In summary, collagen fiber dispersion in cervical tissue of successfully detorted uterine torsion affected buffaloes suggested their role in cervical dilatation, whereby, intracervical hyaluronidase was able to enhance the cervical tissue collagen dispersion.

Correlation of Post-Treatment Biopsy and Fluorodeoxyglucose PET/CT Findings Following Definitive Chemotherapy and Radiation for Cervical Cancer

Concurrent chemotherapy and external beam radiation with brachytherapy boost is considered standard of care for treatment of locally advanced cervical cancer. Post treatment surveillance fluorodeoxyglucose positron emission tomography/computed tomography (FDG-PET/CT) imaging is recommended 3-6 months after completion of therapy. Persistent FDG avidity is commonly observed at the cervix on surveillance scans, indeterminate for residual malignancy versus post treatment inflammatory change. To better understand the clinical significance of residual hypermetabolic activity, we performed an analysis of surveillance FDG-PET/CT images, correlating findings with post treatment cervical biopsy results after completion of therapy. The medical records of all patients treated with definitive chemotherapy and radiation for locally advanced (FIGO IB3 to IVA) cervical cancer at a single academic medical center between 2017 and 2022 were reviewed. Patients were treated with curative intent using a combination of pelvic external beam radiation with concurrent cisplatin chemotherapy followed by intracavitary or combined intracavitary/interstitial brachytherapy boost. Patients underwent FDG-PET/CT imaging at 3-6 months upon completion of therapy, followed by biopsy either for suspected residual disease or equivocal findings on physical exam or FDG-PET/CT scans. Biopsies were obtained within 100 days of FDG-PET/CT imaging (median, 14 days). Twenty-three patients met inclusion criteria. One patient was excluded from analysis due to lack of FDG avidity on her pre-treatment FDG-PET/CT. Among the 22 evaluable scans, median pre-treatment FDG- PET/CT SUVmax of the cervix was 16.3 (range, 4.9-55). Median post-treatment SUVmax was 4.4 (range, 2.1-7.4). Median change in SUVmax from pre-treatment to post-treatment was 11.8 (range, 1.1 to 50.6). Twenty patients had a post-treatment SUVmax of greater than 3. Among post treatment scans, eight showed no residual uptake; two showed uptakes equal to the liver; eleven showed uptakes moderately higher than liver; and one showed uptake markedly increased relative to liver. None of the biopsy specimens showed evidence of residual malignancy, instead showing fibroinflammatory necrosis and reactive changes. None of the patients developed clinical evidence of disease progression at the cervix within one year of treatment completion. Residual hypermetabolic activity at the cervix is common following definitive chemotherapy and radiation for cervical cancer. This is unlikely to represent residual viable malignancy. Post treatment FDG-PET/CT imaging remains valuable for detecting locoregional or distant disease progression/persistence following therapy.

MGMT Methylation Is Associated with Human Papillomavirus Infection in Cervical Dysplasia: A Longitudinal Study.

Cervical premalignancy/malignancy, as detected by cervical cytology or biopsy, can develop as a result of human papillomavirus (HPV) infection. Meanwhile, DNA methylation is known to be associated with carcinogenesis. In this study, we thus attempted to identify the association between MGMT methylation and persistent HPV infection using an Epi-TOP MPP assay. Integrative analysis of DNA methylation was carried out here using longitudinal cervical cytology samples of seven patients with atypical squamous cells of undetermined significance/low-grade squamous intraepithelial lesion (ASC-US/LSIL). Then, a gene expression analysis using the longitudinal cervical cytology samples and a public database (The Cancer Genome Atlas (TCGA)) was performed. Upon comparing the ASC-US or LSIL samples at the 1st collection and the paired samples at the 2nd collection more than 6 months later, we found that they became hypermethylated over time. Then, using the longitudinal data, we found that the MGMT methylation was associated with HPV infection. Moreover, TCGA dataset revealed an association between downregulated MGMT mRNA expression and poor overall survival. This decreased MGMT mRNA expression was observed to have an inverse relationship with MGMT methylation levels. In this study, we found that the MGMT methylation level could potentially serve as a valuable prognostic indicator for the transition from ASC-US/LSIL to cervical cancer.

Correlation of immediate prevalence of cervical squamous cell precancers and cancers with HPV genotype and age in women with LSIL cytology: A retrospective analysis of 1617 cases.

To evaluate the immediate risk of cervical squamous cell precancers and cancers in women with low-grade squamous intraepithelial lesion (LSIL) cytology according to different high-risk human papillomavirus (hrHPV) results and age stratification. The study included 1617 women with LSIL cytology and underwent simultaneous Aptima HPV genotyping (E6/E7 mRNA test) followed by cervical biopsy. Among 1317 hrHPV positive cases, other 11 types of hrHPV were the most frequent (68.8%), followed by HPV16 (11.1%), HPV18/45 (4.1%), and HPV16/HPV18/45 (0.5%). Compared to other groups, HPV18/45 positive group and other 11 types of hrHPV group showed significantly higher prevalence of intraepithelial neoplasia grade (CIN)1 (p < .0001), while HPV16 positive and HPV16/HPV18/45 dual positive groups showed significantly higher prevalence of CIN2/3 (p < .0001). In addition, hrHPV positive, 25-39 years-old age group showed a significantly higher prevalence of CIN1 (p = .032) than the other age groups. Furthermore, CIN1 prevalence was significantly higher in patients under 40 or 50 years of age than in those over 40 or 50 years of age (p = .005 and p = .011, respectively). However, there was no significant difference among the different age groups in CIN2/3 prevalence in women with LSIL cytology. In southern Chinese women population, LSIL cytology carries very low immediate risk of high-grade squamous intraepithelial lesions (HSIL) (CIN2/3) in general. However, HPV16 positive and HPV16/HPV18/45 dual positive indicated a higher immediate risk of high-grade squamous intraepithelial lesions (HSIL) (CIN2/3). Age is not an immediate risk factor in this patient population for high-grade squamous lesions or SCC. These results are similar to data from cytology laboratories in the United States and other international settings, therefore strongly support the usage of ASCCP guidelines in this patient population.