Cervical Intraepithelial Neoplasia grade 2 biopsy: Do p16INK4a and Ki-67 biomarkers contribute to the decision to treat? A cross-sectional study

ABSTRACT BACKGROUND: Managing cervical intraepithelial neoplasia grade 2 (CIN2) is challenging, considering the CIN2 regression rate, perinatal risks associated with excisional procedures, and insufficient well-established risk factors to predict progression. OBJECTIVES: To determine the ability of p16INK4a and Ki-67 staining in biopsies diagnosed with CIN2 to identify patients with higher-grade lesions (CIN3 or carcinoma). DESIGN AND SETTING: Cross-sectional study conducted at a referral center for treating uterine cervical lesions. METHODS: In 79 women, we analyzed the correlation of p16INK4a and Ki-67 expression in CIN2 biopsies with the presence of a higher-grade lesions, as determined via histopathology in surgical specimens from treated women or via two colposcopies and two cytological tests during follow-up for untreated women with at least a 6-month interval. The expression of these two biomarkers was verified by at least two independent pathologists and quantified using digital algorithms. RESULTS: Thirteen (16.8%) women with CIN2 biopsy exhibited higher-grade lesions on the surgical excision specimen or during follow-up. p16INK4a expression positively and negatively predicted the presence of higher-grade lesions in 17.19% and 86.67% patients, respectively. Ki-67 expression positively and negatively predicted the presence of higher-grade lesions in 40% and 88.24% patients, respectively. CONCLUSIONS: Negative p16INK4a and Ki67 immunohistochemical staining can assure absence of a higher-grade lesion in more than 85% of patients with CIN2 biopsies and can be used to prevent overtreatment of these patients. Positive IHC staining for p16INK4a and Ki-67 did not predict CIN3 in patients with CIN2 biopsies.

Tumor risk markers in recurrent respiratory papillomatosis

Abstract Objectives: This study aims to investigate the pattern of immunoexpression of proteins p16, p53 and Ki-67 in RRP, as well as to evaluate its influence on the number of surgeries that patients have undergone to date and to analyze the benefit of immunohistochemistry in this disease. Methods: Clinical-demographic data and tumor samples were obtained from 33 patients with RRP. The expression of proteins p16, p53 and Ki-67 was analyzed by immunohistochemical method. Results: Most patients had already undergone more than one surgery. The p16 marker was negative in 24.2% of the cases, with little positivity in 27.3% of the cases, moderate in 36.4% and intense in 12.1%. The p53 marker was positive in all cases, with little immunoexpression in 39.4% of cases, moderate in 36.4% and intense in 24.2%. The Ki-67 marker showed nuclear positivity in all lesions, although in varying degrees, with a mean proliferative index ±SD (standard deviation) of 51.7±26. Conclusions: The papillomatous lesions had varying degrees of immunoexpression of proteins p16, p53 and Ki-67, but no specific immunohistochemical pattern was observed. It was found, with statistical difference, that the number of surgeries was higher in cases with greater intensity of p53 expression, without correlation with the other markers. The benefit of immunohistochemistry in recurrent respiratory papillomatosis may lie in the prognostic assessment. However, further studies are needed to evaluate the use of this technique for this purpose. Level of evidence:: 4.

Evaluation of variable p16 immunostaining patterns, Ki-67 indices and HPV status in cervical SILs and squamous cell carcinomas: An institutional experience

Objective: The present study evaluates the immunoexpression of p16 and Ki-67 in cervical squamous intraepithelial lesion (SIL) and carcinomas and correlates their expression with clinicopathological features and HPV-DNA status. Material and Methods: A total 36 included cases of SIL and squamous cell carcinoma (SCC) were subjected to p16 and Ki-67 immunostaining. p16 staining was evaluated depending on grading, distribution, localization pattern, intensity and IHC score. Ki-67 expression was graded based on percentage of positive cells. Results: Incidence of HSIL and SCC cases was found to be significantly increased with parity > 5. p16 grade III diffuse nucleocytoplasmic immunostaining was observed in 62.5% LSIL, 80% HSIL and 87% SCC cases. Significant association of p16 staining intensity, IHC score and Ki-67 indices was noted with increasing grades of SILs and carcinomas. Conclusion: Our experience indicates that a combination of p16 and Ki-67 immunostaining may be useful to determine the severity of dysplastic change.

Prognostic significance of p16, VEGF, and p53 expression patterns, Krouse stage, and smoking status in sinonasal inverted papillomas

Aim: The aim of our study is to analyze the staining models for VEGF, p16, and p53, as well as to understand the biology of inverted papilloma caused by smoking. Materials and Methods: Thirty-one cases, diagnosed with sinonasal inverted papilloma between 2015 and 2019, were included. Demographic data such as age and gender, admission symptoms of the patients, and anatomical location, stage, surgical technique, and recurrence information were obtained from clinical follow-up files. Immunohistochemical staining for p16, VEGF, and p53 were performed on patient materials. Results: In our study, the female to male ratio was 9.33 with an average age of 53.137 ± 13.96 years. Of the patients, 17 were nonsmokers and 14 were smokers. No significant relationship was found between smoking status and relapse and dysplasia. In contrast, a significant relationship between the Krouse stage and dysplasia (P = 0.005) was observed. A similar significant relationship was observed between p16 immunohistochemical expression and dysplasia (P = 0.030). On the other hand, VEGF and p53 immunohistochemical expressions were not significantly related with dysplasia and recurrence. Conclusions: Inverted papillomas are benign tumors that clinically give symptoms similar to nasal polyps. However, recurrence and malignant transformation potential exist and the factors causing this risk are not clearly identified. In our study, no malignant transformation was observed in patients who were admitted to our hospital.

Expression of p16 in pterygium and its relation with epithelial dysplasia and possible etiologic role of HPV

Context: p16 is an important tumor suppressor gene and responsible for regulating the cell cycle. Diffuse positivity with p16 in the cervix and head/neck carcinomas can be regarded as a surrogate marker of the presence of high-risk human papillomavirus (HPV). Aim: The aim of our study was to search the existence of p16 expression in pterygium. We also analyzed the association of p16 expression with epithelial dysplasia and HPV expression. Subjects and Methods: The study enrolled 75 cases of pterygium. The conjunctival tissues of 10 patients excised by the strabismus surgery were used as control group. All of the slides were stained with p16 via the immunohistochemical method. Results: 49 (65%) of pterygiums showed low-grade epithelial dysplasia. None of the control groups showed dysplasia. Positive expression of p16 in patient group was significantly higher (P < 0.001). Staining percentage (SP) of p16 was between 0 and 26% in pterygium; mean SP was 5.1%. There was no staining in the control group. A total of 59 (72%) pterygium cases were positive with p16. Appoximately 42 of 49 (85%) cases with dysplasia showed p16 staining. There was a significant relation between dysplasia and positive expression of p16 (P < 0.001). Conclusions: P16 is significantly expressed in pterygium and correlated with epithelial dysplasia. Furthermore, the existence of p16 expression suggests that HPV is a possible ethiological factor in pterygium. We think that examination of p16 expression and analysis of HPV DNA in p16 positive cases can help us to understand the etiopathogenesis of the disease better.

A COMPARATIVE STUDY ON CERVICAL CANCER SCREENING METHOD: IS IMMUNO-MARKER A BETTER OPTION?

Cervical cytology by Papanicolaou (PAP) staining has been the backbone of primary screening of cervical cancer. For low resource countries, the major constraints in running a successful screening program are paucity of experienced personnel, requirement for multiple visits, economic considerations and inherent attributes of the test including a low sensitivity and specificity. The present study was designed to compare the efficacy of commonly available screening tests and feasibility of immuno-markers (p16 & Ki-67) as a primary screening tool. This was a cross-sectional analytical study. 100 patients were approached and agreed to participate in this study. They underwent cervical screening by multiple modalities including PAP smear, Liquid based cytology (LBC), HPV- DNA testing, cytology + HPV- DNA/p16/ Ki-67 and dual markers. Screening test results were compared with histopathology and statistical analysis done. The sensitivity and specificity of conventional cytology was 61.11% and 70%; LBC: 88.88% and 50%; Cytology + HPV DNA: 94.44% and 50%; Cytology + p16: 88.88% and 60%; Cytology + Ki-67: 88.88% and 100%; P16+Ki67: 88.88% and 60%. Combining Ki- 67 with Conventional cytology improved specificity and positive predictive value of cervical cancer screening.

Clinicopathological study in 28 cases of oral basaloid squamous cell carcinomas / 北京大学学报(医学版)

OBJECTIVE@#To investigate the clinicopathologic features and prognostic factors in oral basaloid squamous cell carcinoma.@*METHODS@#Retrospective analysis of oral basaloid squamous cell carcinomas patients who underwent tumor resection during the period from January 2002 to December 2020 in the authors' hospital, especially the clinicopathologic characteristics of 28 cases with confirmed diagnosis and follow-up data. Immunohistochemistry was performed to define the helpful markers for differentiation diagnosis. The factors influencing the prognosis were evaluated based on Kaplan-Meier method.@*RESULTS@#The tongue and mouth floor (11 cases, 39.3%) were the most frequently involved sites, followed by gingiva (6 cases, 21.4%), buccal (5 cases, 17.9%), palate (4 cases, 14.3%), and oropharynx (2 cases, 7.1%). The majority of basaloid squamous cell carcinomas were in advanced stage, with 12 cases in stage Ⅱ and 16 cases in stages Ⅲ-Ⅳ. Twelve of 28 patients were identified to have cervical lymph node metastasis, which was confirmed by histopathological examination. The incidence rate of lymph node metastasis was 42.9% (12/28). Nine tumors recurred, with one metastasized to the lung. At the meantime, the 28 conventional squamous cell carcinomas were matched with the same stage, among which 13 cases were identified with cervical lymph node metastasis. The incidence rate of lymph node metastasis was 46.4% (13/28). Five cases recurred, with two cases that metastasized to the lung and one to the brain. The 5-year overall survival rates of the basaloid squamous cell carcinoma and conventional squamous cell carcinoma patients were 54.6% and 53.8%, respectively. Histopathologically, basaloid cells consisted of tumor islands without evident keratinization but frequently with comedo-like necrosis within the tumor islands. CK5/6 and P63 exhibited strongly positive in all the 28 cases, whereas neuroendocrine markers, CgA and Syn, were negative. Eight cases positively expressed P16; one case showed focal SOX10 positive but CK7 negative.@*CONCLUSION@#The majority of oral basaloid squamous cell carcinomas present in advanced stage with a high tendency to lymph node metastasis, but the overall survival rates are not significantly different from conventional squamous cell carcinomas matched with the same stage. The human papilloma virus (HPV), as HPV-positivity rate is high, correlates to good prognosis. In addition, CK7 & SOX10 immunohistochemistry could contribute to differential diagnosis for basaloid squamous cell carcinoma with solid adenoid cystic carcinoma.

Comparison of values of immunocytochemical P16/Ki-67 double staining, P16 INK4α single staining and high-risk human papillomavirus testing in screening of high-grade cervical lesions / 肿瘤研究与临床

Objective:To investigate the screening values of immunocytochemical P16/Ki-67 double staining, P16 INK4α single staining and high-risk human papillomavirus (HR-HPV) testing for high-grade cervical lesions. Methods:The clinical data of 622 patients who underwent cervical thin-layer liquid-based cytology (TCT) and HR-HPV testing in General Hospital of Taiyuan Iron and Steel (Group) Co., Ltd. from March 2019 to July 2021 were retrospectively analyzed. The remaining cytological specimens were detected by P16/Ki-67 double staining and P16 INK4α single staining. Among them, 334 patients with TCT results suggesting atypical squamous cell of undetermined significance (ASCUS) and above and HPV-positive underwent colposcopy pathological biopsy. Using pathological results as reference, the positive predictive value, sensitivity, specificity and accuracy of P16/Ki-67 double staining, P16 INK4α single staining and HR-HPV testing for screening of high-grade squamous intraepithelial neoplasia (HSIL) and cervical cancer were compared. Results:Taking the results of histopathology as references, combined with the results of TCT, 31 of 622 patients were HSIL, of which 22 (71.0%) were positive for P16/Ki-67 double staining, 23 (74.2%) were positive for P16 INK4α single staining, and 25 (80.6%) were positive for HR-HPV testing; 4 cases were cervical cancer, and the positive rates of the three detection methods were all 100.0% (4/4). Among 622 patients, the positive rates of P16/Ki-67 double staining, P16 INK4α single staining and HR-HPV testing for screening of HSIL and cervical cancer were 13.99% (87/622), 25.40% (158/622) and 21.38% (133/622); the positive predictive values were 29.89%, 17.09% and 21.08%; the accuracies were 91.19%, 78.94% and 83.28%; the specificities were 89.77%, 77.98% and 82.46%; the sensitivities were 74.29%, 77.14% and 82.86%. The positive rate, positive predictive value, specificity and accuracy of P16/Ki-67 double staining were higher than those of P16 INK4α single staining and HR-HPV testing, and the differences were statistically significant ( z values were -5.062 and -3.418, 2.328 and 2.450, 5.436 and 3.570, 6.043 and 4.161, all P < 0.05); the sensitivity of HR-HPV testing was higher than that of P16/Ki-67 double staining and P16 INK4α single staining, but the differences were not statistically significant ( z values were -0.890 and 1.017, both P > 0.05). Conclusions:HR-HPV testing is more suitable for primary cervical lesion screening; P16/Ki-67 double staining can be used as a potential combined cell screening tool or an effective triage tool; P16 INK4α single staining has certain limitations.

Expression and clinical significance of estrogen receptor, progesterone receptor, p53 and p16 protein in endometrial carcinoma / 中国基层医药

Objective:To investigate the expression and clinical significance of estrogen receptor (ER), progesterone receptor (PR), p53 and p16 protein in endometrial carcinoma.Methods:The endometrial tissue of 57 patients with endometrial carcinoma who received surgery in The First People's Hospital of Chuzhou between January 2017 and May 2021 was harvested as the study group. The normal endometrial tissue of 30 patients with endometrial hyperplasia was selected as the control group. Envision immunohistochemical staining was performed to determine the expression of ER, PR, p53 and p16 protein in endometrial tissue and analyze their expression with clinical pathological characteristics.Results:ER, PR, p16 protein expression rates in the endometrial tissue in the study group were 70.2%, 61.4%, 38.6%, respectively, which were significantly lower than 90.0%, 86.7%, 93.3% in the control group ( χ2 = 4.36, 5.98, 24.09, all P < 0.05). p53 expression rate in the endometrial tissue was significantly higher in the study group than that in the control group (52.6% vs. 13.3%, χ2 = 12.75, P < 0.001). ER and PR expression were significantly different between endometrial carcinoma patients with lymph node metastasis and those without and among those with different histological grades and those at different pathological stages (all P < 0.05). There was no significant difference in p53 protein expression among patients with different pathological stages of endometrial carcinoma, between patients who suffered endometrial carcinoma at different ages, and between patients with different degrees of myometrial invasion (all P > 0.05). p16 protein expression rate differed among patients with different pathological stages of endometrial carcinoma, among those with different histological grades and between patients with different degrees of myometrial invasion (all P < 0.05). There was no significant difference in p16 protein expression rate between endometrial carcinoma patients with lymph node metastasis and those without ( P > 0.05). Conclusion:Abnormal expressions of ER, PR, p53 and p16 protein in endometrial tissue may be related to the occurrence, development and transformation of the disease. Combined detection of ER, PR, p53 and p16 protein is helpful for the clinical diagnosis, treatment and prognosis assessment of endometrial carcinoma.

PARP-1 participates in regulation of cell cycle signaling in the hydroquinone-induced TK6 malignant transformation / 中国职业医学

@#To investigate the expression of polyadenosine diphospho-ribose polymerase 1 (PARP-1) and p16/ retinoblastoma (Rb) protein in hydroquinone (HQ)-induced TK6 cells and their regulatory mechanisms. Methods According to the 2×2 factorial design model, TK6 cells were divided into PBS-TK6 group and HQ-TK6 group based on HQ exposure, and then sub-divided into non-DOX intervention subgroup and DOX intervention subgroup based on DOX intervention, a total of four groups. The PBS-TK6 group was treated with phosphate buffer saline, and the HQ-TK6 group was treated with HQ at a final concentration of 20.0 μmol/L. The non-DOX intervention subgroup was added with 0.05% dimethyl sulfoxide; and the DOX intervention subgroup was added with PARP-1 agonist DOX at a final concentration of 0.5 μmol/L. The distribution of cell cycle was detected by flow cytometry. The protein expression of p16/Rb, cyclin D1 (cyclinD1), multifunctional protein E2 transcription factor 1 (E2F1), Rb, and p-Rb were detected by Western blot, and the level of p16 ribosylation was detected by immunofluorescence and immunoprecipitation. Results Compared with the PBS-TK6 group, the cell cycle distribution percentage in G0/G1 phase and the relative expression levels of p16 proteins were decreased in the cells of the HQ-TK6 group (all P<0.05). The cell cycle distribution percentage in S phase and the relative expression levels of cyclinD1 and p-Rb proteins were up-regulated (all P<0.05). Compared with the non-DOX intervention group, the cell cycle distribution percentage in G0/G1 and G2/M phases and the relative expression level of p16 protein increased in the DOX intervention group (all P<0.05). The percentage of cells in S phase and the relative expression levels of cyclinD1 and p-Rb proteins were down-regulated (all P< 0.05). The results of interaction effect analysis showed that compared with the non-DOX PBS-TK6 cells, the relative expression levels of Rb and E2F1 protein in the DOX PBS-TK6 cells intervention group were down-regulated (all P<0.05). The relative expression level of Rb protein in non-DOX HQ-TK6 cell group was down-regulated (P<0.05), and the relative expression of E2F1 protein was up-regulated (P<0.05). Compared with DOX PBS-TK6 cell group, the relative expression level of Rb protein in DOX HQ-TK6 cell group was down-regulated and that of E2F1 protein was up-regulated (all P<0.05). Compared with the non-DOX HQ-TK6 cell group, the relative expression level of Rb protein in the DOX HQ-TK6 cell group was up-regulated and that of E2F1 protein was down-regulated (all P<0.05). Conclusion PARP-1 participates in cell cycle regulation by regulating the p16/Rb signaling pathway in TK6 cells.

Presence of Human Papillomavirus and Epstein-Barr Virus in Squamous Lesions of the Tongue

ABSTRACT Background: Several studies have suggested a possible role of human papillomavirus (HPV) and Epstein-Barr virus in the pathogenesis of oral premalignant lesions. This study aimed to investigate the correlation between squamous dysplasia of the tongue and expression of p16 and Ki67 immunohistochemically as well as HPV genotypes with real-time polymerase chain reaction (PCR). Methods: Twenty-three tongue biopsies were stained immunohistochemically for p16, Epstein- Barr virus and Ki67 and real-time PCR and chromogenic in-situ hybridization for HPV. Results: Dysplasia was diagnosed in 16 of 23 cases without invasive carcinoma and suspicious for dysplasia (n=17) and HPV infection (n=6). These were subjected to chromogenic in-situ hybridization for HPV DNA (HPV-III family 16). There was no immunoreactivity for Epstein-Barr virus. p16 was positive in 4/16 (25%) of dysplastic lesions. One lesion was positive for HPV by chromogenic in-situ hybridization, and one case was positive by real-time PCR for HPV. Conclusion: This evidence suggested that HPV infection but not Epstein-Barr virus infection plays a role in pathogenesis of squamous dysplasia localized tongue.

Presença do papilomavirus humano tipo 16 e expressão gênica da proteína p16ink4a e oncoproteína e7 no carcinoma colorretal / Hpv16 and expression of protein p16ink4a and e7 oncoprotein in colorectal carcinoma

RESUMO - INTRODUÇÃO: O papilomavírus humano (HPV) é agente das doenças sexualmente transmissíveis de maior prevalência no mundo que estão associadas ao câncer do colo do útero e canal anal. A ação do HPV na carcinogênese colorretal não está ainda estabelecida. OBJETIVO: Estudar a eventual correlação entre a presença do HPV tipo 16 e a expressão gênica da proteína p16INK4a e da oncoproteína E7 de HPV e de seus níveis no tecido do carcinoma colorretal. METODOS: Estudo retrospectivo caso-controle de 79 doentes com carcinoma colorretal divididos em dois grupos: HPV presente e HPV ausente. Foi realizada reação em cadeia da polimerase (PCR), além da hibridização do tipo dot blot para o HPV 16 e o HPV 18 Amostras do tecido colorretal também foram submetidas ao estudo imuno-histoquimico para avaliar o nível tecidual das proteínas E7 e p16INK4a. RESULTADOS: O HPV foi identificado em 36 (45,6%) casos. Não houve diferença significante entre os grupos quanto ao sexo (p=0,056), idade (p=0,1), localização cólica e/ou retal (0,098) e presença do HPV. A expressão gênica da oncoproteína E7 de HPV estava presente em 3,12% dos casos (p=0,9) e a expressão da proteína p16INK4a foi observada em 46,3% (p=0,27) dos indivíduos com detecção do HPV. CONCLUSÃO: A expressão gênica e os níveis teciduais da oncoproteína E7 e da proteína p16INK4a encontrados nos pacientes positivos para o HPV sugerem a ausência de atividade oncogênica do HPV tipo 16 no carcinoma colorretal.

ABSTRACT - BACKGROUND: Human papillomavirus (HPV) is the agent of the most prevalent sexually transmitted diseases in the world associated with cervix and anal canal cancer. The action of HPV on colorectal carcinogenesis is not yet established. OBJECTIVE: This research aimed to study the possible correlation between the presence of HPV16 and the gene expression of p16INK4a protein and HPV E7 oncoprotein and their levels in colorectal carcinoma tissue. METHODS: A retrospective case-control study of 79 patients with colorectal carcinoma was divided into two groups: HPV-positive and HPV-negative. The polymerase chain reaction was performed, in addition to dot-blot hybridization for HPV16 and HPV18. Colorectal tissue samples were also subjected to immunohistochemical study to assess the tissue level of E7 and p16INK4a proteins. RESULTS: HPV was identified in 36 (45.6%) cases. There was no significant difference between groups regarding gender (p=0.056), age (p=0.1), colic and/or rectal location (0.098), and presence of HPV. Gene expression of HPV E7 oncoprotein was present in 3.12% of cases (p=0.9), and p16INK4a protein expression was observed in 46.3% (p=0.27) of those selected with HPV detection. CONCLUSION: Gene expression and tissue levels of E7 oncoprotein and p16INK4a protein found in HPV-positive patients suggest the absence of HPV16 oncogenic activity in colorectal carcinoma.

Is there correlation between human papillomavirus (hpv) and esophageal epidermoid carcinoma? / Há correlação entre o papilomavírus humano (hpv) e o carcinoma epidermoide de esôfago?

ABSTRACT Background: Currently, persistent human papillomavirus (HPV) infection has been related in some geographic regions as a risk factor for esophageal squamous cell carcinoma. It results in the immunoexpression of the p16 protein, which has been used as marker of the oncogenic lineage by this etiological agent. Aim: To correlate epidemiological aspects of esophageal squamous cell carcinoma with the prevalence of HPV infection. Methods: Fifty-eight cases were analyzed and submitted to histopathological and immunohistochemical analysis by p16. Results: Of the 58 cases evaluated, 40 were men and 18 women, with a mean age of 63.2 years. p16 immunoexpression was positive in 46.55%. Conclusion: The prevalence of HPV infection is high in esophageal squamous cell carcinoma presenting in almost half of the cases (46.55%), without gender differentiation.

RESUMO Racional: Atualmente a infecção persistente pelo papilomavírus humano (HPV) tem sido relacionada em algumas regiões geográficas como fator de risco para o carcinoma epidermoide do esôfago. Ela resulta na imunoexpressão da proteína p16, que tem sido utilizada como marcadora da linhagem oncogênica por este agente etiológico. Objetivo: Correlacionar aspectos epidemiológicos do carcinoma epidermoide do esôfago com a prevalência de infecção pelo HPV. Métodos: Foram analisados 58 casos buscando-se perfil epidemiológico dos pacientes, com suas peças submetidas à análise histopatológica e imunoistoquímica pelo p16. Resultado: Dos 58 casos avaliados, 40 eram homens e 18 mulheres, com idade média de 63,2 anos. A imunoexpressão pelo p16 foi de 46,55%. Conclusão: A prevalência de infecção pelo HPV é alta no carcinoma epidermoide de esôfago apresentando-se em quase a metade dos casos (46,55%), sem diferenciação de idade quanto aos gêneros.

Diagnostic value of combined detection of p16 and human papillomavirus typing in cervical intraepithelial neoplasia / 肿瘤研究与临床

Objective:To explore the application value of combined detection of p16 and human papillomavirus (HPV) typing in the diagnosis of cervical intraepithelial neoplasia (CIN).Methods:A total of 8 346 patients aged between 25 years old and 65 years old at Baoji Central Hospital of Shaanxi Province from February 2019 to February 2020 were selected. There were 2 882 patients with cervical lesions diagnosed by colposcopy biopsy. Patients were divided into the different groups based on the age range, and then the condition of HPV infection in all age groups was analyzed. Taking biopsy as the gold standard and according to the pathological results, the detection rate of p16 and HPV typing and the diagnostic value of the single and combined detection in CIN were also analyzed.Results:The age group with the highest positive rate of p16 and HPV was 31-40 years old [47.42% (1 014/2 427) and 36.84% (894/2 427), respectively], followed by 41-50 years old group [30.15% (907/2 942) and 28.11% (827/2 942)], and there were statistically significant differences in positive rate of p16 and HPV in all age groups (all P < 0.05). Among 2 882 patient with cervical lesions diagnosed by pathological examination, there were 2 572 cases (89.24%) of p16 positive, and 2 169 cases (75.26%) of HPV positive. With the disease progression of cervical lesions, the positive rate of p16 and HPV was gradually increased, and the positive rate of p16 of inflammation, CINⅠ, CINⅡ, CIN Ⅲ, cervical squamous cell carcinoma (SCC) was 11.68% (23/197), 94. 85% (1 105/1 165), 93.57% (771/824), 96.76% (538/556), 96.43% (135/140), respectively; the positive rate of HPV was 17.77% (35/197), 77.60% (904/1 165), 80.22% (661/824), 80.40% (447/556), 87.14% (122/140), respectively, and HPV infection was mostly HPV16/18 infection type with the disease progression of cervical lesions. The sensitivity, specificity, positive predictive value and negative predictive value in detecting CIN of HPV was 75.26%, 81.13%, 67.78% and 86.14%, respectively; the sensitivity, specificity, positive predictive value and negative predictive value in detecting CIN of p16 was 89.24%, 84.74%, 75.51% and 93.72%, respectively; the diagnostic efficacy of p16 was higher than that of HPV in detecting CIN, and the difference was statistically significant ( P < 0.05). The sensitivity, specificity, positive predictive value and negative predictive value of HPV combined with p16 in detecting CIN was 94.10%, 91.33%, 85.12%, 96.71%, which were higher compared with those of single detection (all P < 0.05). Conclusions:HPV infection mainly occurs in women aged 31-40 years old followed by 41-50 years old, and the infected population of CIN tends to be younger. p16 is superior to HPV in detecting the diagnostic efficacy of CIN; combined detection of p16 and HPV can increase the sensitivity and specificity, reduce the rate of misdiagnosis, and can play a key clinical value in early diagnosis and treatment of CIN.

Detection of p16/Ki-67 in women with abnormal cytological results and its diagnostic value for cervical intraepithelial neoplasia II+ grade / 国际生物医学工程杂志

Objective:To explore the detection capability of p16/Ki-67 double staining technique in women with various abnormal thinprep cytologic test (TCT) results and its diagnostic value for cervical intraepithelial neoplasia Ⅱ+ grade (CIN2+).Methods:A total of 225 women with abnormal TCT results, i.e. the atypical squamous cells of undetermined significance(ASC-US), in the Tianjin Central Hospital of Gynecology Obstetrics, Nankai University Affiliated Maternity Hospital from December 2018 to December 2019 were enrolled. p16/Ki-67 double staining were detected and compared with the high risk human papillomavirus (HR-HPV) and pathological results.Results:The positive rates of p16/Ki-67 double staining increased with cytologic and pathologic categories. For diagnosis of CIN2+, p16/Ki-67double staining (90.1%) was less sensitive than HR-HPV testing (98.2%)( P<0.05), but the specificity of p16/Ki-67 double staining (58.8%) was significantly higher than HR-HPV(21.6%) ( P<0.001). Conclusions:Compared with HR-HPV, p16/Ki-67 double staining has better effect on diagnosing CIN2+. p16/Ki-67 double staining can be considered as triaging method for management of ASC-US and LSIL patients, significantly reduce the colposcopy referral rate (nearly 50%), which has high clinical application value.

Evaluation of the P16INK4a Expression in Cervical Biopsy Specimen SBMCH

In early 1980 human papillomavirus (HPV) were the risk factor and most commonly affects younger women. Many test have developed since then and among that a biomarker test system have developed and clinically evaluated. P16INK4a is used as an important marker for indicating neoplastic transformation for cervical dyplasia. This study was done to evaluate the P16INK4a expression in cervical biopsy in 50 cases. Two cases were identified a P16INK4a positive and remaining 48 didn’t show P16INK4a expression proving the hypothesis that p16INK4a is capable of showing the dysplasia positive cases

Assessment of HPV 16, HPV 18, p16 expression in advanced stage laryngeal cancer patients and prognostic significance / Avaliação da expressão do HPV 16, HPV 18, e p16 em pacientes com câncer de laringe em estágio avançado e significado prognóstico

Abstract Introduction: Human papilloma virus is an etiological risk factor for a subset of head and neck squamous cell carcinomas. HPV has been proven to be a powerful prognostic biomarker for oropharyngeal cancer, but its role in the larynx has not been explored in depth. The developmental mechanisms of laryngeal carcinomas are quite complex and controlled by various factors. Smoking and alcohol are most important risk factors. Recent studies indicate that HPV infection also plays an important role in larynx carcinomas. HPV related laryngeal carcinomas especially occur at the supraglottic region of larynx. Objective: We aimed to determine the frequency of HPV/protein16 positivity in patients with laryngeal carcinoma and association of HPV and/or p16 positivity with variables such as age, sex, smoking habits, tumor localization, lymph node metastasis, recurrence and survival in advanced stage laryngeal carcinoma in our study. Methods: This retrospective study included 90 patients with advanced laryngeal carcinoma. The Control group was 10 normal larynx mucosa specimens. The presence of HPV was investigated polyclonally by polymerase chain reaction, and protein16 with immunohistochemical method. In HPV positive cases, the presence of HPV types 16, 18 were evaluated by polymerase chain reaction. Demographic features of patients were noted. Patient survival and association with HPV/protein16 was determined. Results: Polyclonal HPV positivity was detected in 11 (12.2%) of 90 cases. Out of these 11 cases, HPV 16 was positive in 6, HPV 18 in 4, and both HPV 16 and 18 were positive in 1. In 18 (20%) of the cases, p16 was positive. Six of the cases (6.6%) had both HPV and protein16 positivity. In cases where protein16 alone or HPV and protein16 were co-positive, alcohol use was less and the tumor was found more likely to be localized in the supraglottic area. These ratios were statistically significant. Supraglottic localization of tumor was determined to be increased in protein16 positive cases. The correlation between protein16 positivity and supraglottic area location was determined to be statistically significant (p= 0.011). 55.6% of protein16 positive cases was located in the supraglottic region, 33.3% was glottic and 11.1% was transglottic. Although life expectancy over 5 years were numerically higher in HPV and protein16 positive cases, this was not found to be statistically significant. There was no statistically significant relationship between HPV positivity and mean age, differentiation, smoking and alcohol use, tumor progression, lymph node metastasis, localization, recurrence, cause of mortality and treatment methods in our study. The mean follow-up period of our patients was 6.7 years. Conclusion: The close relationship between HPV and oropharyngeal squamous cell carcinoma could not be shown in larynx malignancy in many studies, including our study. Our findings support a limited role of HPV in laryngeal carcinogenesis. Protein16 is not a reliable surrogate for HPV status in laryngeal cancers and is not a predictor of laryngeal cancer survival. Supraglottic localization of tumor was determined to be increased in protein16 positive cases. The correlation between protein16 positivity and supraglottic area location was determined to be statistically significant. There is a need for more populated clinical trials, where neoplastic proliferation is better demonstrated and the accuracy of the results obtained is supported by different techniques.

Resumo Introdução: O papilomavírus humano é um fator de risco etiológico para um subconjunto de carcinoma espinocelular de cabeça e pescoço. Tem sido demonstrado que o HPV é um poderoso biomarcador prognóstico para o câncer de orofaringe, mas seu papel na laringe ainda não foi explorado em profundidade. Os mecanismos de desenvolvimento dos carcinomas de laringe são bastante complexos e controlados por vários fatores. Tabagismo e álcool são os fatores de risco mais importantes. Estudos recentes indicam que a infecção pelo HPV também desempenha um papel importante nos carcinomas da laringe. Os carcinomas laríngeos relacionados ao HPV ocorrem especialmente na região supraglótica. Objetivo: Nosso objetivo foi determinar a frequência da positividade para o HPV / proteína 16 em pacientes com carcinoma da laringe e a associação da positividade para o HPV e /ou proteína 16 com variáveis como idade, sexo, tabagismo, localização do tumor, metástase linfonodal, recidiva e sobrevivência de carcinoma da laringe em estágio avançado em nosso estudo. Método: Este estudo retrospectivo incluiu 90 pacientes com carcinoma laríngeo avançado. O grupo controle incluiu 10 amostras de mucosa laríngea normal. A presença de HPV foi inves-tigada por anticorpo policlonal através de reação de polimerase em cadeia e a proteína 16 por método imunohistoquímico. Nos casos positivos para o HPV, a presença dos tipos 16 e 18 do foi avaliada por reação de polimerase em cadeia. As características demográficas dos pacientes foram observadas. A sobrevida dos pacientes e a associação com HPV / proteína 16 foram determinadas. Resultados: A positividade com anticorpo policlonal do HPV foi detectada em 11 (12,2%) dos 90 casos. Desses 11 casos, o HPV 16 foi positivo em 6, o HPV 18 em 4 e o HPV 16 e 18 foram positivos em 1. Em 18 (20%) dos casos, a proteína 16 foi positiva. Seis dos casos (6,6%) apresentaram positividade para HPV e proteína16. Nos casos positivos apenas para a proteína 16 ou quando HPV e a proteína 16 foram co-positivos, a ingestão de álcool foi menor e o tumor apresentou maior probabilidade de estar localizado na área supraglótica. Essas proporções foram estatisticamente significantes. A localização supraglótica do tumor foi maior em casos positivos para proteína 16. A correlação entre positividade para proteína 16 e localização da área supraglótica foi estatisticamente significante (p = 0,011). Dos casos positivos para proteína 16, 55,6% foram supraglóticos, 33,3% glóticos e 11,1% transglóticos. Embora a expectativa de vida acima de 5 anos tenha sido numericamente maior nos casos positivos para HPV e proteína 16, isso não foi estatisticamente significante. Não houve relação estatisticamente significante entre positividade do HPV e média de idade, diferenciação, tabagismo e uso de álcool, progressão tumoral, metástase linfonodal, localização, recidiva, causa de mortalidade e métodos de tratamento em nosso estudo. O período médio de seguimento de nossos pacientes foi de 6,7 anos. Conclusão: A estreita relação entre HPV e carcinoma espinocelular orofaríngeo não pôde ser demonstrada na laringe em muitos estudos, inclusive no nosso estudo. Nossos achados confirmam um papel limitado do HPV na carcinogênese da laringe. A proteína 16 não é um substituto confiável para o status do HPV nos cânceres de laringe e não é preditor da sobrevida do câncer de laringe. A localização supraglótica do tumor foi maior em casos positivos para proteína16. A correlação entre positividade para proteína 16 e localização na área supraglótica foi determinada como estatisticamente significante. Há necessidade de ensaios clínicos com amostras maiores, nos quais a proliferação neoplásica seja melhor demonstrada e a precisão dos resultados obtidos seja apoiada por diferentes técnicas.

Molecular Quantification of Epstein-Barr Virus Nuclear Antigen Gene and DNA Methylation Patterns of Human Tumor Suppressor Genes p16 and E-cadherin in Relation to Infection with Helicobacter pylori as Early Prognostic Biomarker for Gastric Tumorgenesis in Patients from Baghdad

Objectives: To investigate the association between EBV viral infectious dose, infection with EBV and/or H. pylori and histologically different gastric diseases and cancer. Also, to study the association between EBV and/or H. pylori infection with DNA methylation patterns of human tumor suppressor genes p16 and CDH1 in progression of primary gastric diseases to neoplasia. Methods: The current prospective cross-section study included a total of 94 GTPs taken from patients suffering from gastro-duodenal manifestations recruited to Gastro-Endoscopy Department at Gastroenterology and Hepatology Teaching Hospital, Baghdad, Iraq, from November 2017 to October 2018. Two gastric tissue biopsies (GTBs) were collected from those patients. Genomic DNA was extracted from fresh GTPs. Direct molecular identification of H. pylori in extracted DNA was performed by amplification of species-specific urea. Identification of Epstein-Barr nuclear antigen 1(EBNA1) in extracted DNA was performed using nested PCR. DNA samples positive to EBNA1 were submitted for viral load estimation using quantitative real time PCR. Methylation patterns of p16 and CDH1 promoters were detected in modified DNA samples by sodium sulfate using MS-PCR. Results: Of total samples, 39 (41.5%) of DNA samples were positive for H. pylori and 18 (19.15%) DNA samples were positive for EBNA-1. Studying EBV load, 8/23DNA samples were showed infectious dose of EBV. Studying methylation patterns of p16 and CDH-1 promoters, 21/42 and 19/42 DNA samples were provide results for MSPPCR, respectively. Conclusion: Epstein-Barr virus and H. pylori infection may have a synergistic effect in developing different gastric diseases and that enable the clinician to choose the suitable treatment regime.

Comparison of senescence progression in mesenchymal cells from human umbilical cord walls measured by immunofluorescence and flow cytometry of p16 and p21 / Comparação da progressão da senescência em células mesenquimais de parede de cordão umbilical humano medida por imunofluorescência e citometria de fluxo de p16 e p21

ABSTRACT Objective To follow the expansion of mesenchymal stem cells from umbilical cords by two classic senescence markers, p16 (INK4A) and p21 (CDKN1A), using practical, fast, and less expensive methods than the gold standard Western blotting technique, to evaluate its applicability in the laboratory. Methods Mesenchymal stem cells from umbilical cords were isolated from Wharton's jelly and, after quality control, morphological and immunophenotypic characterization by flow cytometry, were expanded in culture until coming close to cell cycle arrest (replicative senescence). Results A comparison was made between young cells, at passage 5, and pre-senescent cells, at passage 10, evaluating the protein expression of the classic cell senescence markers p16 and p21, comparing the results obtained by Western blotting with those obtained by flow cytometry and indirect immunofluorescence. Conclusion Follow-up of cell cultures, through indirect p16 immunofluorescence, allows the identification of mesenchymal stem cells from umbilical cord cultures at risk of reaching replicative senescence.

RESUMO Objetivo Acompanhar a expansão de células-tronco mesenquimais de cordão umbilical por dois marcadores clássicos de senescência, p16 (INK4A) e p21 (CDKN1A), usando métodos práticos, rápidos e com custo menor do que a técnica padrão-ouro de Western blotting, para avaliar sua aplicabilidade em laboratório. Métodos Células-tronco mesenquimais de cordão umbilical foram isoladas da geleia de Wharton e, após controle de qualidade e caracterização morfológica e imunofenotípica por citometria de fluxo, foram expandidas em cultura, até chegarem próximas à parada do ciclo celular (senescência replicativa). Resultados Foi feita a comparação entre células jovens, na passagem 5, e células pré-senescentes, na passagem 10, avaliando a expressão proteica dos marcadores clássicos de senescência celular p16 e p21, comparando os resultados obtidos por Western blotting com os obtidos por citometria de fluxo e imunofluorescência indireta. Conclusão O seguimento de culturas celulares, por meio da imunofluorescência indireta de p16, permite identificar as culturas de células-tronco mesenquimais de cordão umbilical em risco de atingirem a senescência replicativa.

Guilu Erxian Glue () Inhibits Chemotherapy-Induced Bone Marrow Hematopoietic Stem Cell Senescence in Mice May via p16 / 中国结合医学杂志

OBJECTIVE@#To evaluate the effect of Guilu Erxian Glue (, GEG) on cyclophosphamide (CTX)-induced bone marrow hematopoietic stem cells (HSCs) senescence in mice and explore the underlying mechanism.@*METHODS@#The H@*RESULTS@#Compared with the model group, GEG increased cell viability as well as proliferation (P<0.05 or P<0.01) and reduced β -gal expression. Furthermore, GEG significantly decreased the expressions of p16@*CONCLUSION@#GEG can alleviate CTX-induced HSCs senescence in mice, and the p16