Detection of high-risk HPV 16 genotypes in cervical cancers using isothermal DNA amplification with electrochemical genosensor.

Cervical cancer emerges as the third most prevalent types of malignancy among women on a global scale. Cervical cancer is significantly associated with the persistent infection of human papillomavirus (HPV) type 16. The process of diagnosing is crucial in order to prevent the progression of a condition into a malignant state. The early detection of cervical cancer through initial stage screening is of the utmost significance in both the prevention and effective management of this disease. The present detection methodology is dependent on quantitative polymerase chain reaction (qPCR), which necessitates the use of a costly heat cycler instrument. In this study, we report the development of an electrochemical DNA biosensor integrated with an isothermal recombinase polymerase amplification (RPA) reaction for the detection and identification of the high-risk HPV-16 genotype. The electrochemical biosensor exhibited a high degree of specificity and sensitivity, as evidenced by its limit of detection (LOD) of 0.23 copies/µL of HPV-16 DNA. The validity of this electrochemical platform was confirmed through the analysis of 40 cervical tissues samples, and the findings were consistent with those obtained through polymerase chain reaction (PCR) testing. Our straightforward electrochemical detection technology and quick turnaround time at 75 min make the assay suitable for point-of-care testing in low-resource settings.

Role of Human Papilloma Virus 16/18 In Laryngeal Carcinoma with Correlation to the Expression of Cyclin D1, p53, p16 and EGFR.

Objective: To investigate the role of Human Papilloma Virus (HPV) (16/18) in relation to the molecular genetic mechanisms of Cyclin D1, p53, p16, and Epidermal growth factor receptor (EGFR) in Laryngeal Squamous Cell Carcinoma (LSSC). Methods: A cross-sectional study of 88 (Formalin-fixed Paraffin Embedded) FFPE laryngeal biopsies were done at Basic Medical Sciences Institute, Jinnah Postgraduate Centre, Karachi from 2010 to 2019 with the application of Polymerase chain reaction (PCR) for HPV 16/18and Immuno-histochemical staining for molecular genetic expression of proteins, Cyclin D1, p53, p16, and EGFR. Results: Out of 88 cases of Laryngeal Squamous Cell Carcinoma (LSSC) there was female preponderance. Mean age of the participants was found as 50.7±12.8 years. High risk HPV 16/18 was positive in 28 cases (31.8%), largely related to Grade-II and Grade-III. Immunohistochemically, Cyclin D1 (87.5%) appeared as the most important driver mutation followed by p16 (86.4%), EGFR (65.9%), and, p53 was positive in (61.4%) of cases. Conclusion: The role of high-risk HPV 16/18 is concurred in the present study strongly in correlation to p16 as a surrogate marker. Moreover, the other driver mutations of Cyclin D1, p53, and EGFR are also implicated as cumulative molecular events in tumor progression as mostly seen in higher Grades.

T helper cell and regulatory T-cell related cytokines changes in vaginal lavage fluid of patients with high risk-human papilloma virus 16 positive and the predictive effect on the development of cervical neoplasms / 肿瘤研究与临床

Objective:To investigate the changes of T helper cell (Th), regulatory T-cell (Treg cell) related cytokines in vaginal lavage fluid of patients with high risk-human papilloma virus 16 (HR-HPV16) positive and its predictive effect on the development of cervical neoplasms.Methods:A total of 200 cases of HR-HPV16 positive patients who admitted to Xingtai People's Hospital from January 2022 to December 2022 were selected as the experimental group. According to the results of pathological examination, all patients in the experimental group were divided into non pathological group (78 cases), low grade squamous intraepithelial lesion (LSIL) group (49 cases), high grade squamous intraepithelial lesion (HSIL) group (39 cases) and cervical cancer group (34 cases); and 100 healthy people undergoing the physical examination in the same period were taken as the healthy control group. Enzyme-linked immunosorbent assay (ELISA) double-antibody sandwich method was used to detect the levels of interleukin (IL)-2, IL-4, IL-6, IL-10, IL-12, IL-17, tumor necrosis factor α (TNF-α), interferon γ (IFN-γ) and transforming growth factor β (TGF-β) in vaginal lavage fluid of patients in different groups. Multivariate logistics regression was used to analyze the risk factors of cervical cancer, and a nomogram model was established. The receiver operating characteristics (ROC) curve was drawn with pathological results as the gold standard, and the area under the curve (AUC) was calculated to evaluate the predictive ability of the nomogram model.Results:The levels of IL-6, IL-10, IL-17, TNF-α, TGF-β in vaginal lavage fluid of patients in the experimental group were higher than those in the healthy control group, while the levels of IL-2, IL-12 and IFN-γ in the experimental group were lower than those in the healthy control group, and the differences were statistically significant (all P < 0.05); the difference in IL-4 level of both groups was not statistically significant ( P > 0.05). There were statistically significant differences in IL-6, IL-10, IL-17, TNF-α, TGF-β, IL-2, IL-12 and IFN-γ among non pathological group, LSIL group, HSIL group and cervical cancer group (all P < 0.05); the levels of IL-6, IL-10, IL-17, TNF-α, TGF-β in cervical cancer group were the highest, the levels of IL-2, IL-12 and IFN-γ were the lowest; the level of IL-4 in non pathological group, LSIL group, HSIL group and cervical cancer group had no statistically significant difference ( P > 0.05). Logistics regression analysis showed that low IL-2, high IL-4, high IL-6, high IL-10, low IL-12, high IL-17, high TNF-α, low IFN- γ and low TGF-β expressions in vaginal lavage fluid of patients with HR-HPV16 positive were independent risk factors for the development of cervical cancer (all P < 0.05). The results of nomogram analysis showed that IL-6, IL-10, IL-17, TNF-α, TGF-β in vaginal lavage fluid were the factors predicting the development of cervical cancer in HR-HPV16 positive patients. The ROC curve analysis showed that the AUC of nomogram model in predicting the development of cervical cancer in HR-HPV16 positive patients was 0.945 (95% CI 0.901-0.988), and the predictive efficacy was good. Conclusions:Th and Treg cell related cytokines levels in vaginal lavage fluid of patients with HR-HPV16 positive show pathological changes in cervical cancer patients and the above indicators have a high value in predicting the development of cervical cancer.

A novel multi-epitope vaccine of HPV16 E5E6E7 oncoprotein delivered by HBc VLPs induced efficient prophylactic and therapeutic antitumor immunity in tumor mice model.

Human papilloma virus type 16 (HPV16) is the most prevalent etiologic agent associated with cervical cancer, and its early proteins E5, E6 and E7 play important roles in cervical epithelium transformation to cervical intraepithelial neoplasia and even cervical cancer. Hence, these oncoproteins are ideal target antigens for developing immunotherapeutic vaccines against HPV-associated infection and cervical cancer. Currently, multi-epitope vaccines have been a promising strategy for immunotherapy for viral infection or cancers. In this study, the E5aa28-46, E6aa37-57 and E7aa26-57 peptides were selected and linked to form a novel multi-epitopes vaccine (E765m), which was inserted into the major immune dominant region (MIR) of hepatitis B virus core antigen (HBc) to construct a HBc-E765m chimeric virus-like particles (cVLPs). The immunogenicity and immunotherapeutic effect of the cVLPs vaccine was evaluated in immunized mice and a tumor-bearing mouse model. The results showed that HBc-E765m cVLPs elicited high E5-, E6- and E7- specific CTL and serum IgG antibody responses, and also relatively high levels of the cytokines IFN-γ, IL-4 and IL-5. More importantly, the cVLPs vaccine significant suppressed tumor growth in mice bearing E5-TC-1 tumors. Our findings provide strong evidence that this novel HBc-E765m cVLPs vaccine could be a candidate vaccine for specific immunotherapy in HPV16-associated cervical intraepithelial neoplasia or cervical cancer.

Survival-related indicators ALOX12B and SPRR1A are associated with DNA damage repair and tumor microenvironment status in HPV 16-negative head and neck squamous cell carcinoma patients.

OBJECTIVES: To investigate prognostic-related gene signature based on DNA damage repair and tumor microenvironment statue in human papillomavirus 16 negative (HPV16-) head and neck squamous cell carcinoma (HNSCC). METHODS: For the RNA-sequence matrix in HPV16- HNSCC in the Cancer Genome Atlas (TCGA) cohort, the DNA damage response (DDR) and tumor microenvironment (TM) status of each patient sample was estimated by using the ssGSEA algorithm. Through bioinformatics analysis in DDR_high/TM_high (n = 311) and DDR_high/TM_low (n = 53) groups, a survival-related gene signature was selected in the TCGA cohort. Two independent external validation cohorts (GSE65858 (n = 210) and GSE41613 (n = 97)) with HPV16- HNSCC patients validated the gene signature. Correlations among the clinical-related hub differentially expressed genes (DEGs) and infiltrated immunocytes were explored with the TIMER2.0 server. Drug screening based on hub DEGs was performed using the CellMiner and GSCALite databases. The loss-of-function studies were used to evaluate the effect of screened survival-related gene on the motility of HPV- HNSCC cells in vitro. RESULTS: A high DDR level (P = 0.025) and low TM score (P = 0.012) were independent risk factors for HPV16- HNSCC. Downregulated expression of ALOX12B or SPRR1A was associated with poor survival rate and advanced cancer stages. The pathway enrichment analysis showed the DDR_high/TM_low samples were enriched in glycosphingolipid biosynthesis-lacto and neolacto series, glutathione metabolism, platinum drug resistance, and ferroptosis pathways, while the DDR_high/TM_low samples were enriched in Th17 cell differentiation, Neutrophil extracellular trap formation, PD - L1 expression and PD - 1 checkpoint pathway in cancer. Notably, the expression of ALOX12B and SPRR1A were negatively correlated with cancer-associated fibroblasts (CAFs) infiltration and CAFs downstream effectors. Sensitivity to specific chemotherapy regimens can be derived from gene expressions. In addition, ALOX12B and SPRR1A expression was associated with the mRNA expression of insulin like growth factor 1 receptor (IGF1R), AKT serine/threonine kinase 1 (AKT1), mammalian target of rapamycin (MTOR), and eukaryotic translation initiation factor 4E binding protein 1 (EIF4EBP1) in HPV negative HNSCC. Down-regulation of ALOX12B promoted HPV- HNSCC cells migration and invasion in vitro. CONCLUSIONS: ALOX12B and SPRR1A served as a gene signature for overall survival in HPV16- HNSCC patients, and correlated with the amount of infiltrated CAFs. The specific drug pattern was determined by the gene signature.

Human papilloma virus infection of uterine cervix and spectrum of cervical pathology in human immunodeficiency virus/AIDS.

BACKGROUND: Human papilloma virus (HPV) is one of the most common causes of sexually transmitted viral diseases worldwide. High-risk HPV types such as HPV16 and 18 are known to cause cervical dysplasia and carcinoma. In human immunodeficiency virus (HIV)-positive individual, chance of HPV coinfection and risk of cervical dysplasia/carcinoma have been found to be significantly more than in HIV-negative individuals. AIM: In this institution-based, cross-sectional, observational study, we aim to find out the relationship of HPV infection of the uterine cervix with cervical dysplasia and neoplasia in HIV-infected/AIDS patients. MATERIALS AND METHODS: Conventional Pap smears were taken from HIV-infected individuals admitted in the department of gynecology and obstetrics and reported by the Bethesda system. A second sample was sent to the virology unit of ICMR for detection and typing of HPV. Control samples were taken from HIV-negative individuals. RESULTS: Fifty HIV-positive patients were included in this study. On cervical Pap smear examination, 32 cases were cytologically benign and 18 cases showed atypical cytomorphology. Twenty-four cases were HPV positive, among which 16 were cytologically atypical and 8 were benign. HPV 16 was the most common subtype (50%) followed by HPV 18 (37.5%) and others (12.5%) in HIV-positive patients. Chance of cervical dysplasia increased with age independent of HIV infection and with progressive lower CD4 count. Koilocytosis was a significant predictor of HPV infection. Majority of patients were asymptomatic. Peak incidence of HPV infection occurred in reproductive age group (20-40 years). The association between HIV and HPV coinfection (P = 0.002) and between HPV infection and cytology atypia (P < 0.0001) was statistically significant. CONCLUSION: Present study highlights the necessity of routine cervical Pap smear screening in HIV infected reproductive age-group women. Early detection enables dysplasia to revert or be effectively managed.

[Performance of high-risk HPV typing test in early diagnosis of cervical cancer].

Objective: To evaluate the performance of High-risk HPV typing detection in cervical cancer screening. Methods: A total of 3 231 women were recruited as the subjects of cervical cancer screening from Jiyuan city of Henan provinces from June to July 2017. All women underwent HPV DNA test. The women with cytological examination ≥ASCUS or cytological examination negative and HPV 16/18 positive underwent colposcopy biopsy and pathological examination. Pathological diagnosis was used as the gold standard. Sensitivity, specificity, positive predictive value (PPV), negative predictive value(NPV) as well as corresponding 95% confidence interval (CI) of HR-HPV and HPV 16/18 were calculated. Results: The mean age of 3 231 subjects selected in this study was 46.84±10.00 (21-64) years old. 524 subjects had the positive results of HR-HPV, including 91 of HPV16 and 15 of HPV18. Pathological test result CIN2+ was the gold standard. The sensitivity and specificity of HR-HPV for cervical precancer lesions screening were 93.75 (95%CI: 79.85-98.27) and 84.56% (95%CI: 83.26-85.77), respectively. To compared with HR-HPV, HPV16/18 had low sensitivity (65.63%, 95%CI: 48.31-79.59)and higher specificity (97.44%, 95%CI: 96.83-97.93). After age stratification by age 30 and 45, the sensitivity of HPV 16/18 was same with HR-HPV (100%, 95%CI: 34.24-100.00), the specificity of HPV 16/18 was higher than HR-HPV (98.71%, 95%CI: 96.27-99.56 vs 84.48%, 95%CI: 79.27-88.58) in<30 age group.The sensitivity of HR-HPVin 30-45 and ≥45 age group were higher than HPV16/18 (85.71%, 95%CI: 48.69-97.43 vs 71.43%, 95%CI: 35.89-91.78, 95.65%, 95%CI: 79.01-99.23 vs 60.87%, 95%CI: 40.79-77.84), but the specificity werelower than HPV16/18 (86.89%, 95%CI: 84.58-88.90 vs 98.51%, 95%CI: 97.51-99.11、83.49%, 95%CI: 81.81-85.04 vs 96.80%, 95%CI: 95.94-97.48). Conclusions: HR-HPV detection has relatively high sensitivity and specificity in cervical cancer screening. For >30 years old women, HR-HPV is more recommended in cervical cancer screening. Therefore, HR-HPV detection is an effective method for cervical cancer screening.

Reprogrammed CRISPR-Cas13a targeting the HPV16/18 E6 gene inhibits proliferation and induces apoptosis in E6-transformed keratinocytes.

The long-lasting infection of high-risk type (type 16 or type 18) human papillomavirus (HPV) is the main cause of gynecological and urinary malignancies. Given the high mortality rate after surgery, the development of a new molecular therapy would be of value to clinicians. The aim of the present study was to achieve targeted inactivation of the viral E6 gene in keratinocytes using the reprogrammed CRISPR-Cas13a system. To accomplish this, a reprogrammed CRISPR-Cas13a system, targeting both the HPV16/18 E6 genes was constructed using a guide RNA expressing vector. The expression levels of E6 protein were measured using western blot analysis after transfection of the vector into E6-transformed keratin oocytes. Cell proliferation was analyzed using CCK-8 assays and cell apoptosis was evaluated using Hoechst 33258 staining and ELISAs examining caspase-3 levels. The results indicated that both the HPV16/18 E6 genes can be inactivated using the CRISPR-Cas13a system. Furthermore, silencing E6 inhibited cell proliferation (14±1.8% on average) and induced apoptosis (80.2±3.2% on average) in E6-transformed keratinocytes but not in normal keratinocytes. In conclusion, results of the present study demonstrate that the reprogrammed CRISPR-Cas13a system has the potential for inactivating HPV E6 gene functions.

Luciferase-Based Detection of Antibodies for the Diagnosis of HPV-Associated Head and Neck Squamous Cell Carcinoma.

Point-of-care tests are needed for the screening of head and neck squamous cell carcinoma (HNSCC) and other malignancies. Luciferase immunoprecipitation systems (LIPS), employing light-emitting proteins, were used to examine serum antibodies against several cancer-associated targets in blood donor controls and subjects with colon cancer (CC) and HNSCC. The assessment of antibodies against the wild type p53 tumor antigen showed that approximately 25% of the CC and 20% of the HNSCC patients were seropositive. In addition, humoral responses against two p53 mutants, p53-R175H and p53-R273H, generally tracked the antibody responses seen against wild type p53. Analysis of antibodies against highly specific biomarkers of HPV-16-associated malignancy, E2, E6, and E7 oncoproteins, revealed no seropositivity in blood donors and CC patients. However, 45% (9/20) of the HNSCC patients showed E6 seropositivity, which overlapped all the detectable E2 (40%; 8/20) and E7 seropositive subjects (35%; 7/20). Using neodymium magnets, ultrarapid LIPSTICKS testing of HPV-16 E6 antibodies in <60 s per HNSCC sample demonstrated almost the same diagnostic performance (40% sensitivity and 100% specificity) as LIPS testing in 2.5 h. While additional improvements and standardization are needed, these results highlight the possibility of using these approaches for the diagnosis of HPV-16-associated HNSCC.

HPV16 E7 increases COX-2 expression and promotes the proliferation of breast cancer.

Breast cancer remains the leading cause of mortality worldwide. Human papilloma virus 16 (HPV16) may serve a function in the pathogenesis and development of breast cancer. However, the detection rate of HPV16 in breast carcinoma may vary by region. In the present study, the expression of HPV16 E7 in paraffin-embedded tissues from patients with breast cancer from North China was detected. Additionally, the molecular mechanisms underlying the function of HPV16 E7 in the proliferation of breast cancer cells were examined. The results demonstrated that the DNA of HPV16 E7 was detected in 30.5% of the samples, and that HPV16 E7 promoted the proliferation of breast cancer cells in vitro and in vivo. Additionally, HPV16 E7-mediated proliferation of breast cancer cells was suppressed in response to treatment with cyclooxygenase-2 (COX-2)-specific small interfering RNA and celecoxib. The results of the present study revealed that HPV16 E7 may promote the proliferation of breast cancer cells by upregulating COX-2, suggesting that COX-2 may be a potential therapeutic target for HPV16 E7-mediated progression of breast cancer.

Human papilloma virus 16/18: Fabricator of trouble in oral squamous cell carcinoma.

AIM: To find out the association between Human Papilloma Virus (HPV) genotypes 16/18 in Pakistani patients with oral squamous cell carcinoma (OSCC). METHODS: DNA from oral rinse of 300 subjects was taken. The subjects included 100 cases with OSCC and 200 controls. Samples were analyzed by both conventional and real time PCR using "HPV consensus Gp5+/Gp6+ and HPV 16, 18 specific primers". RESULTS: Out of 300 persons, 74/300 (25%) were found to be infected with HPV: "46/100(46%) from cases and 74/200(14%) from controls". The distribution was: HPV16, 6/300 (8%): 4/100 (9%) from OSCC group and 2/200 (8%) from controls while HPV 18 was 9/300(12%): 5/100(11%) from cases and 4/200(16%) from controls. Out of 300 subjects, 26(35%) were infected by "both HPV 16/18 (23(50%) from cases and 3(12%) from controls". Persons who were infected with HPV 16&18 had higher chances to develop OSCC as compared to those who didn't have HPV 16/18 (AOR: 21.4, 95% CI: 5.73 - 80.8). CONCLUSION: The exposure to high risk strains of Human papilloma virus (16/18) in combination can be fabricotor of trouble (p<0.001, Adjusted odds ratio; 21.42) in OSCC.

Expression of IL-17 and estrogen and its receptor in HPV16 continuously infected patients / 局解手术学杂志

Objective To understand the role of estrogen,estrogen receptor and IL-17 in human papillomavirus (HPV) infection and clearance.Methods We selected the clinic or the hospitalized patients in People' s Hospital of Pudong New District hospital with HPV negative or single-HPV16 positive during the period of August 2014 to January 2017.Cervical exfoliated cells were harvested by Thinprep cytologic test (TCT).The reverse spot hybridization technique was carried out for HPV subtype analysis.RT-PCR was employed for HPV16 DNA viral load.The levels of E2 and IL-17 were detected by ELISA.Immunohistochemical was used to detect ER expression.The experiments were repeated every three months.According to the difference in the outcome of human papilloma virus 16 infection,the experiment was divided into three groups.The first group was HPV negative,which was negative for one year.The second group was HPV16 positive,but within one year the difference was gradually cleared.The third group was HPV16 positive,but the HPV16 persistently existed after one year.Results The results of the last test were compared with the first test results.We found that There was no significant difference among the three groups in the expression of E2 and ER.Also,there was no significant difference between the HPV negative group and the HPV16 persistently infected group in variation of IL-17 concentration.But the difference between the HPV16 cleared group and the anyone of the two groups above was significantly.The variation of IL-17 concentration was significantly higher in the HPV16 cleared group.Conclusion In normal cytology,and only HPV16 infected stage,endogenous E2 and ER of cervix may not play a role in HPV infection,or they may have little effect on HPV infection.Local immune factor IL-17 plays an important role in the process of HPV16 removal,and the increase of IL-17 concentration can help to eliminate HPV16.

Differential proteins among normal cervix cells and cervical cancer cells with HPV-16 infection, through mass spectrometry-based Proteomics (2D-DIGE) in women from Southern México.

BACKGROUND: Cervical cancer (CC) is the fourth most common cancer in women worldwide with an estimated 528,000 new cases in 2012. The same year México had an incidence of 13,960 and a mortality of 4769 cases. There are several diagnosis methods of CC; among the most frequents are the conventional Pap cytology (Pap), colposcopy, and visual inspection with acetic acid (VIA), histopathological examination, tests of imaging and detection of high-risk papilloma virus (HR-HPV) with molecular tests (PCR, hybridization, sequencing). Proteomics is a tool for the detection of new biomarkers that can be associated with clinical stage, histological type, prognosis, and/or response to treatment. In this study we performed a comparative analysis of CC cells with normal cervical cells. The proteomic analysis was carried out with the fluorescent two-dimensional electrophoresis (2D-DIGE) technique to subsequently identify differential protein profiles using Decyder Software, and the selected proteins were identified by Mass Spectrometry (MALDI-TOF). RESULTS: The proteins that showed an increased expression in cervical cancer in comparison with normal cervix cells were: Mimecan, Actin from aortic smooth muscle and Lumican. While Keratin, type II cytoskeletal 5, Peroxiredoxin-1 and 14-3-3 protein sigma showed a decrease in their protein expression level in cervical cancer in comparison with normal cervix cells. CONCLUSIONS: Thus, this study was successful in identifying biomarker signatures for cervical cancer, and might provide new insights into the mechanism of CC progression.

Correlation between Severity of Cervical Lesions and Methylation of Human Papilloma Virus Type 16 DNA / 中国医科大学学报

Objective To quantifiably measure the methylation frequency of 18 CpG sites in the 3′region of L1 gene and long control region(LCR) gene of HPVl6 DNA,and study the relationship between HPVl6 DNA methylation and severity of cervical lesions. Methods A total of 10 cases Normal/low?grade squamous intraepithelial lesion(Normal/LSIL),10 cases of high?grade squamous intraepithelial lesion(HSIL),and 10 cases of cervical cancer(CC)were recruited for the study. The relationship between severity of cervical lesions and HPV16 DNA methylation was analyzed by bisultlte?pyrosequencing. Results The methylation rate was highest in Normal/LSIL at position 7 089 located in 3′?L1,followed by CC. The low?est was found in HSIL. The difference in methylation percentage among the three lesions was significant(P=0.006). In 7 134,the proportion meth?ylation was also different among three groups(P=0.01),difference in methylation percentage between Normal/LSIL and CC,as well as Normal/LSIL and HSIL was significant(P=0.038,0.017). Conclusion The methylation status of CpG sites 7 089 and 7 134 in the 3′region of L1gene is asso?ciated with the severity of cervical disease. The quantification of HPV DNA methylation can be used for cervical disease screening in clinical samples.

Role of human papilloma virus-16 in the pathogenesis of oral lichen planus--an immunohistochemical study.

BACKGROUND: Oral lichen planus (OLP) is a common chronic inflammatory immune-mediated disease with an aetiopathogenesis associated with cell-mediated immunological dysfunction. It is possible that oral mucosal viral infections, including human papilloma virus-16 (HPV-16) infection, may have a causative role in OLP pathogenesis. AIM: To assess the prevalence of HPV-16 in histopathologically diagnosed specimens of OLP and to evaluate whether any clinical features (such as the localisation of specimens) or the age or gender of patients, are correlated with the presence of this virus. MATERIALS AND METHODS: This study was conducted on 30 specimens with a histopathological diagnosis of OLP, using the immunohistochemical marker HPV-16. Thirty normal oral mucosa specimens were also included as controls. Brown nuclear staining was accepted as positive for the HPV-16 antibody. The results were analysed using Fisher's exact test. P values<0.05 were considered to be significant. RESULTS: Significant correlation (P=0.0001) was observed between HPV-16 infection and samples with OLP. No statistical conclusions could be drawn regarding age, gender, localisation and HPV-16 positivity. CONCLUSION: Our study showed that HPV-16 may play a role in the pathogenesis of OLP. Taking into account the oncogenic potential of HPV-16, patients with OLP should be screened for the presence of this virus.

RESEARCH THE EXPRESSION OF HIGH RISK HUMAN PAPILLOMA VIRUS AND P16 PROTEIN IN CERVICAL INTRAEPITHELIAL LESION BY IMMUNOHISTOCHEMISTRY CH ABOUT LAPAROSCOPY ASSISTED VAGINAL HYSTERECTOMY / Шинэ санаа Шинэ нээлт

The persistent high-risk human papilloma virus(HPV) infection is a necessary cause for developing cervical carcinoma. Although carcinogenic HPV types are found in virtually all invasive cancer, with types 16 and 18 being found in approximately 70 percent of cases. High risk HPV types’ Е6 and Е7 oncogenes have a pivotal role in cervical carcinogenesis. The p16, the cyclin-dependent kinase inhibitor and p16 overexpression in cervical neoplasia is a surrogate marker of high risk HPV E7 mediated pRb catabolism reflecting disruption of mechanisms that control cell proliferation and indicating persistent infection with high risk of development of neoplasia.Thus in worldwide p16 had been identified as the novel biomarker in pre-invasive cervical lesions. Objective: For the purpose to detect for cervical cancer risks we examined HPV16/18 and cell cycle protein p16 expression in cervical lesions.A total of 96 specimens enrolled in this study and 50 were diagnosed as LSIL and 46 were diagnosed as a HSIL. To detect HPV16/18 and p16 in cervical lesions used immunohistochemistry. Statistical analysis was performed using SPSS 16.0. Descriptive analysis was performed by Chi- Square test and also determined sensitivity and specificity.Positive stainingfor p16 and HPV16/18 were observed whole cell, within both the nuclear and cytoplasmic subcellular regions by immunohistochemistry. 63% of specimens had only HPV16 infection and 22% of specimens had only HPV18 infection.Also 14% specimens had co-infection with two viral types and 28% specimens had not above two most HPV infection. There were a significant difference for HPV16 positivity (X2 = 4.93, P 0.05) in HSIL and LSIL groups. There were not a difference for p16 in HSIL and LSIL groups.(X2 = 0.23, P > 0.05), respectively.P16, yielding a diagnostic sensitivity for HPV 16/18 were 82% and 30%, specificity for HPV 16/18 were 40% and 80%, respectively. In conclusion it is possible to detect high risk HPV types and persistent infection by immunohistochemistry in cervical intraepithelial squamous cell lesions. There is still critical need to use HPV testing and other molecular surrogate markers of HPV such as p16 in primary screening program.

Significance of detection of suPAR, SCC-Ag and HPV16, 18 in patients with cervical cancer / 肿瘤研究与临床

Objective To evaluate the significance of suPAR,SCC-Ag in plasma and HPV16,18 in cervical secretion for monitoring pathogenetic condition and prognosis in patients with cervical cancer.Methods 206 cervical cancer patients blood and cervical secretion were collected.Plasma level of suPAR and SCC-Ag were measured by enzyme linked immunosorbent assay (ELISA) in health women and patients with cervical cancer.The expression of HPV16,18 of cervical secretion in control group and patients with cervical cancer were detected by fluorescence quantitative PCR.The correlations of the three indexes were analyzed.Results The plasma level of suPAR and SCC-Ag,the expression of HPV16,18 of cervical secretion in cervical cancer patients were obviously higher than those in health controls with statistical significance ((1.072 5±0.305 2) ng/ml vs (0.501 7±0.179 3) ng/ml,(0.980 6±0.162 7) μg/ml vs (0.261 4± 0.006 3) μg/ml and 53.89 ％ (90/167),46.15 ％ (18/39) vs 6.67 ％ (4/60),P ＜ 0.05).There was a positive correlation between plasma suPAR level and SCC-Ag level in invasive carcinoma of cervix patients (r =0.564,P ＜ 0.05).The plasma level of suPAR between in HPV16,18 positive group and in HPV16,18 negative group did not show difference (P ＞ 0.05).Conclusions In invasive carcinoma of cervix patients,there is a positive correlation between plasma suPAR level and SCC-Ag level.But it's not yet to conclude that plasma suPAR level of cervix invasive carcinoma patients is related to infection of HPV16,18.

Human papilloma virus in head and neck squamous cell cancer.

BACKGROUND: Epidemiologic and molecular evidences have established a strong link between high risk types of Human Papilloma Virus and a subgroup of Head and Neck Squamous Cell Carcinomas (HNSCC). We evaluated the frequency of HPV positivity in HNSCC and its relationship to demographic and some risk factor variables in an open case- control study. METHODS: Fourteen recently diagnosed patients with squamous cell cancer of oropharynx, hypopharynx and larynx aged 18-50 years were examined from 2008-2010 in Tabriz, Iran. HPV DNA was extracted from paraffin-embedded blocks of each patient's sample for PCR evaluation. Saliva samples of 94 control cancer-free subjects were collected for DNA analysis. Multivariable logistic regression method was used to calculate odds ratio for case-control comparisons. RESULTS: High risk HPV was detected in 6(42.8%) patients, and 6(5.3%) control subjects which was statistically significant (p<0.0001). HPV-18 was the most frequent type both in the cases and controls. HPV-16 DNA was detected in two patients of the case group, but it was not detected in any of the controls. The relation between demographic and risk factor variables was not statistically significant. CONCLUSION: HPV infection has a significant impact on HNSCC. Despite HPV-16 stronger impact, HPV-18 is more likely to cause malignant degeneration in such cancers amongst some communities. It is vital to introduce and conduct immunization schedules in health care systems to protect communities to some extent.

¿Es el momento de vacunar contra el Virus del Papiloma Humano en Colombia? / Is it time to vaccinate against human papilloma virus in Colombia?

Objetivo: analizar el estado actual de la evidencia de la vacunación para el virus de papiloma humano (VPH) a nivel poblacional teniendo como referente su posible aplicación en Colombia. Metodología: evaluación de estudios fase III sobre las vacunas contra el VPH: estudios de los grupos FUTURE I, II y PATRICIA. Se reflexiona sobre su generalización a nivel poblacional y se analizan factores que limitan su uso. Resultados: la evaluación de estos estudios encuentra limitaciones metodológicas que controvierten la eficacia presentada en las publicaciones de los estudios como son la elección de variables sustitutas inadecuadas, análisis de resultados con variables primarias compuestas que son heterogéneas, análisis por protocolo y no por intención de tratamiento, criterios de inclusión que no sustentan la aplicación poblacional de la vacuna en menores de 15 años y factores que pueden afectar la validez de los estudios como conflictos de intereses y origen del patrocinio. De otra parte, se presentan reportes de complicaciones con la aplicación de la vacuna que brindan una mejor información sobre el verdadero comportamiento de este método de prevención por fuera del diseño experimental. Conclusiones: para el momento de esta revisión no se tiene la suficiente evidencia que sustente la inclusión de la vacuna contra el VPH como programa de vacunación poblacional en Colombia

Objective: analysing current evidence regarding proposals for vaccinating the Colombian population against human papilloma virus (HPV). Methodology: assessing HPV vaccine phase III studies (FUTURE I, II and PATRICIA study groups). Comments are made regarding generalising such vaccination amongst the general population, also taking factors limiting their use into consideration. Results: methodological limitations were found when assessing these studies thereby challenging the importance of the efficacy found in such studies such as the choice of inadequate substitutes, analysing results involving heterogeneous primary compounds, analysis by protocol and not by intention to treat, inclusion criteria which did not underpin a general vaccination programme for girls aged younger than 15 and factors which could affect the validity of the studies such as conflicts of interests and sources of sponsorship. Several complications have been reported regarding how the vaccine has been applied, providing better information about this prevention method’s real behaviour (i.e. when taken beyond its experimental design stage). Conclusions: there was not sufficient evidence to recommend introducing HPV vaccination within Colombian immunisation schedules when this review was made

Induction of SiHa Cells Cells Apoptosis by Nanometer Realgar Suspension and Its Mechanism / 华中科技大学学报（医学）（英德文版）

The effects of nanometer realgar suspension on proliferation and apoptosis of human uterine cervix cancer cell line SiHa cells and oncogenic genes HPV16E6/E7 were investigated. A "micro-jet efflux" strategy was used for the preparation of nanometer realgar suspension. SiHa cells were treated with nanometer Realgar suspension in various concentrations (6.25,12.5,25 and 50mg/L) for different durations (12,24,48 and 72h). The inhibitive effect of nanometer realgar suspension on growth of SiHa cells was detected by MIT method. Special morphological changes of apoptosis were observed by transmission electron microscopy (TEM) and DNA fragments electrophoresis. The apoptotic rate was quantified by flow cytometry (FCM). The expression of HPV16E6/E7 mRNA and protein was assayed by RT-PCR and Western blot respectively. The results showed after being treated with 25-50mg/L nanometer realgar suspension for 48h, the survival rate of SiHa cells was decreased, and apoptotic rate markedly increased in a time- and concentration-dependent manner. TEM and DNA electrophoresis revealed the special morphological changes of apoptosis. The apoptotic rate of SiHa cells treated with nanometer realgar suspension was significantly higher than in the control group (P<0.01), and G0/G1 phase arrest appeared following treatment with nanometer realgar suspension in 25 and 50mg/L for 48h.RT-PCR and Western blot assay indicated that nanometer realgar suspension reduced the HPV16E6/E7 gene expression. Nanometer realgar suspension could inhibit the proliferation and induce apoptosis of SiHa cells. The mechanism may be related to the down-regulation of the HPV16E6/E7 gene expression.