Traditional Chinese Medicine Erhuang Suppository for Treatment of Persistent High-risk Human Papillomavirus Infection and Its Impact on Transcriptome of Uterine Cervix

Objective High-risk human papillomavirus(HR-HPV)infection is the chief cause of cervical intraepithelial neoplasia(CIN)and cervical carcinoma.The Erhuang suppository(EHS)is a traditional Chinese medicine(TCM)prepared from realgar(As_(2)S_(2)),Coptidis rhizoma,alumen,and borneolum syntheticum and has been used for antiviral and antitumor purposes.However,whether EHS can efficiently alleviate HR-HPV infection remains unclear.This study was conducted to evaluate the efficacy of EHS for the treatment of persistent HR-HPV infection in the uterine cervix.Methods In this study,we evaluated the therapeutic efficacy of EHS in a randomized controlled clinical trial with a 3-month follow-up.Totally,70 patients with persistent HR-HPV infection were randomly assigned to receive intravaginal administration of EHS or placebo.HPV DNA,ThinPrep cytologic test(TCT),colposcopy,and safety evaluation were carried out after treatment.Microarray analysis was performed to compare transcriptome profiles before and after EHS treatment.A K14-HPV16 mouse model was generated to confirm the efficiency of EHS.Results After 3 months,74.3%(26/35)of the patients in the treatment group were HPV negative,compared to 6.9%(2/29)in the placebo group.High-throughput microarrays revealed distinct transcriptome profiles after treatment.The differentially expressed genes were significantly enriched in complement activation,immune response,and apoptotic processes.The K14-HPV16 mouse model also validated the remarkable efficacy of EHS.Conclusion This study demonstrated that EHS is effective against HR-HPV infection and cervical lesions.Additionally,no obvious systemic toxicity was observed in patients during the trial.The superior efficacy and safety of EHS demonstrated its considerable value as a potential cost-effective drug for the treatment of HPV infection and HPV-related cervical diseases.

Mother-to-Child Transmission of Human Papillomavirus in Burkina Faso

Introduction: Human papillomavirus (HPV) infection is the most widespread sexually transmitted infection in the world. Today, there is growing evidence that HPV can be transmitted early in life, and one potential route is mother-to-child transmission. Data on this route of HPV transmission are scarce in Africa and particularly in Burkina Faso, where no data on the subject are yet available. The aim of our study was to estimate the rate of mother-to-child transmission of HPV infection and to identify circulating genotypes. Methodology: Cervico-uterine samples were collected from 100 full-term pregnant women and, buccal samples were obtained from their newborns at Hopital Saint Camille de Ouagadougou (HOSCO) by the specialist physician. HPV DNA amplification and genotyping were performed by PCR followed by hybridization using the HPV Direct Flow Chips kit, detecting 36 genotypes including 18 high-risk and 18 low-risk. Results: The prevalence of HPV in newborns was 8% (8/100). Six (6) HPV-positive neonates had HPV-positive mothers, while 2 HPV-positive neonates had HPV-negative mothers. The vertical transmission rate was 26.09% (6/23). Mother-newborn genotypes were concordant. However, the genotype profile of the newborns was more restricted than that of the mothers. Conclusion: HPV DNA was found in 8% of newborns in our study. The genotype profile of the mother-newborn pair was concordant. Asymptomatic HPV infection in a pregnant woman could constitute a risk factor for vertical transmission.

Hysteroscopic cervical biopsy for women with persistent human papillomavirus infection after loop electrosurgical excision procedure: A case report

BACKGROUNDAlmost all cases of cervical cancer can be attributed to human papillomavirus(HPV) infection. The loop electrosurgical excision procedure (LEEP) is widelyused to treat HPV-mediated disease;thus, cervical cancer is highly preventable.However, LEEP does not necessarily clear HPV rapidly and may affect theaccuracy of the results of ThinPrep cytology test (TCT) and cervical biopsy due tothe formation of cervical scars.CASE SUMMARYA 40-year-old woman underwent LEEP for cervical intraepithelial neoplasia grade1 approximately 10 years ago. Subsequent standard cervical cancer screeningsuggested persistent HPV-52 infection, but TCT results were negative. Cervicalbiopsy under colposcopy was performed thrice over a 10-year period, yieldingnegative pathology results. She developed abnormal vaginal bleeding after sexualactivity, persisting for approximately 1 year, and underwent hysteroscopy in ourhospital. Histopathologic evaluation confirmed adenocarcinoma in situ of theuterine cervix.CONCLUSIONPatients with long-term persistent, high-risk HPV infection and negative pathologyresults of cervical biopsy after LEEP are at risk of cervical cancer. Hysteroscopicresection of cervical canal tissue is recommended as a supplement tocervical biopsy because it helps define the lesion site and may yield a pathologicdiagnosis.

Role of viruses in periodontitis:An extensive review of herpesviruses,human immunodeficiency virus,coronavirus-19,papillomavirus and hepatitis viruses

Periodontitis is the inflammation of the supporting structures around the dentition.Several microbial agents,mostly bacteria,have been identified as causative factors for periodontal disease.On the other hand,oral cavity is a rich reservoir for viruses since it contains a wide variety of cell types that can be targeted by viruses.Traditionally,the focus of research about the oral flora has been on bacteria because the most widespread oral diseases,like periodontitis and dental caries,are outcomes of bacterial infection.However,recently and especially after the emergence of coronavirus disease 2019,there is a growing tendency toward including viruses also into the scope of oral microbiome investigations.The global high prevalence of periodontitis and viral infections may point out to a concomitant or synergistic effect between the two.Although the exact nature of the mechanism still is not clearly understood,this could be speculated through the manipulation of the immune system by viruses;hence facilitating the furthermore colonization of the oral tissues by bacteria.This review provides an extensive and detailed update on the role of the most common viruses including herpes family(herpes simplex,varicella-zoster,Epstein-Barr,cytomegalovirus),Human papillomaviruses,Human immunodeficiency virus and severe acute respiratory syndrome coronavirus 2 in the initiation,progression and prognosis of periodontitis.

A study on the cognitive status of human papillomavirus among adult males in Tianjin based on the health belief model

Background:To understand the health beliefs and knowledge of human papillomavirus among adult males in Tianjin.Methods:An online questionnaire survey was conducted from 18 January 2023 to 6 March 2023 using snowball sampling method.Analyze the health belief scores and human papillomavirus(HPV)and HPV vaccine knowledge scores of adult males in Tianjin,and analyze their influencing factors.Results:A total of 388 adult males in Tianjin were surveyed,with an average total score of 3.23±0.04 for their health beliefs.Among them,the average scores for perceived severity,perceived susceptibility,perceived impairment,perceived benefit,and self-efficacy were 3.41±1.05,2.37±1.20,2.96±1.00,3.51±0.90,and 3.36±1.08,respectively.Multiple linear regression analyses showed education was a factor influencing health beliefs.The average total score of knowledge is 64.09±15.62,with 277 people scoring above 60,and a pass rate of 71.4%.Through multiple linear regression analysis,education level,emotional status,whether disease testing has been done,and whether family and friends have been diagnosed with HPV positive are the main influencing factors.Conclusion:The awareness rate of HPV among adult males in Tianjin is still acceptable,but there are still misconceptions.The overall level of health beliefs is moderate,and the perceived susceptibility level is low.It is necessary to strengthen health education on HPV related knowledge for males and improve their cognitive level.

Prevalence of Precancerous Lesions Based on Digital Cervicography with VIA/VILI among Women Positive for High-Risk Human Papillomavirus Serotypes: A Screening Center-Based Study in Cameroon

Background: Since 2021, high-risk Human Papilloma Virus (HR-HPV) testing has been the recommended screening test for cervical cancer for all settings;either used alone in a “test and treat” strategy, or with a triage test, with or without biopsy, before treatment. Cameroon has rolled out immunization against HPV 16 and 18, but studies show a higher prevalence of non-16/18 HR-HPV types. Objectives: Determine the prevalence of precancerous lesions, in women with HR-HPV infection and evaluate association of digital cervicography (DC) VIA/VILI positivity with HPV serotype, as a measure of their contribution to precancer and cancer incidence. Methodology: The study was cross-sectional, descriptive, and analytic. It took place at the Etoug-Ebe and Ekoudoum Baptist Hospitals in Yaoundé, during the period April-September 2022. We reviewed the records of women screened for cervical cancer between February 2020 and December 2021 and evaluated the prevalence of lesions on digital cervicography (DC) with VIA/VILI for women positive for HR-HPV serotypes. The data were analyzed using SPSS version 20.0 for Windows. P values Results: We identified 315 cases with a positive HR-HPV deoxyribonucleic acid (DNA) test, 224 (71.1%) had a DC VIA/VILI triage test done. Of these, 30 (13.4%) women had a positive DC VIA/VILI, with five women (2.2%) having lesions suggestive of cancer. Out of 11 cases positive for HPV 16 alone, 05 (45.5%) had a positive DC VIA/VILI test. Of the 14 cases positive for HPV 18 alone, 03 (21.4%) had a positive VIA/VILI, meanwhile only 19 (10.7%) of the 177 cases positive for non-16/18 HPV had a positive VIA/VILI test. Conclusion: A high proportion of women (13.4%) with HR HPV had a positive DC VIA/VILI, with a significant proportion (2.2%) having lesions suggestive of invasive cervical cancer HR-HPV serotype was associated with DC VIA/VILI positivity;HPV 16 had the strongest association (45.5%), followed by HPV 18 (21.4%), and non-16/18 HR-HPV (10.7%), suggesting a decreasing order of oncogenicity.

Study on the Correlation between Human Papillomavirus and Mycoplasma genitalium Combined with TCT Detection

Objective: This study aims to explore the correlation between human papillomavirus (HPV) and Mycoplasma genitalium (CT) combined with TCT detection in cervical cancer screening. Method: A cross-sectional study design was adopted, and a total of 609 women who came to seek medical treatment were recruited as the study subjects. Combination testing was evaluated on cervical cancer screening by testing the women for HPV, CT with TCT detection and analyzing the relationship of cervical lesions with HPV and CT infection. Results: The study results showed that 21.57% of the subjects were infected with both HPV and CT, and 48.42% of the cases had abnormal TCT results at the same time. Further data analysis showed that HPV infection was significantly associated with abnormal TCT outcomes (p < 0.05), suggesting a possible synergistic effect of the two infections in cervical lesions. The combined sensitivity and specificity of HPV, CT and TCT detection were 21.57% and 48.42%, respectively, which were significantly higher than that of single detection. Conclusion: In summary, the results of this study support the importance of combined HPV, CT, and TCT testing in cervical cancer screening, and propose the hypothesis that combined testing may improve screening effectiveness. However, further large sample studies are needed to confirm this conclusion and explore the prospects of combined testing in clinical practice.

Evaluating the Association between Human Papillomavirus and Vulvar Cancer:A Comprehensive Analysis Using Bradford Hill Criteria

Background:The role of human papillomavirus(HPV)in the development of vulvar cancer(VC)has been widely studied,but findings have been inconsistent.Despite numerous meta-analyses exploring the potential link between HPV and VC,the association remains controversial due to inherent limitations in meta-analytic methods.Objectives:To address this controversy,the study aims to investigate the potential link between HPV and VC using the Bradford Hill criteria,which offer a more comprehensive framework for establishing causation.Methodology:The study began by extracting all relevant studies on the association between HPV and VC from the PubMed database.The potential links were then assessed by examining the data using the major postulates of the Bradford Hill criteria.To ensure the reliability of the findings,the methodologies of the identified studies were critically evaluated to account for possible false-negative and false-positive results.Results:The assessment of previous studies against the Bradford Hill criteria revealed that the major postulates were not fulfilled.Conclusion:Based on the findings,it can concluded that there is no causal association between HPV and VC.

Analysis of Human Papillomavirus Infection in Cervical Exfoliated Cells

To screen patients with early cervical lesions by analyzing the infection of high-risk Human papillomavirus (HR-HPV). Research Methods: The cervical exfoliated cell specimens and their clinical data were collected. The HPV infection types of the collected specimens were detected by fluorescence quantitative PCR, and the correlation between HPV infection and clinicopathological features was analyzed statistically. Results: 725 cases were HR-HPV positive from 2605 cases, including 15 high-risk types of HPV16, 18, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66, 68. Different histological types ranged from NILM to HSIL, and the positive rate of HPV showed an increasing trend with the aggravation of cervical lesions. Conclusion: The positive rate of 15 high-risk HPV types in the collected specimens was 27.8%. Patients with early cervical lesions could be screened for 15 high-risk HPV infection types.

Distribution of High-Risk Human Papillomavirus Genotypes among Women with Colposcopic Diagnosis of Cervical Intraepithelial Neoplasia in Bangladesh

Background: The incidence of cervical cancer is high in Bangladesh and there is a high prevalence of preinvasive lower genital tract disease among women of reproductive age. Persistent high-risk Human Papilloma Virus (HPV) infection is the main underlying cause of cervical cancer and its precursor, cervical intraepithelial neoplasia (CIN). Objective: The aim of the study was to identify the subtypes of high-risk HPV infection among women with the colposcopic diagnosis of cervical intraepithelial neoplasia in Bangladesh. Methods: This cross-sectional observational study was conducted in the colposcopy clinic of Dhaka Medical College Hospital over a six-month period. A total of 100 participants were enrolled. Married women, between 30 - 60 years of age with colposcopically diagnosed cervical intra epithelial neoplasia were enrolled. Women with chronic illness, pregnancy, and women unable to consent were excluded from this study. After counselling, colposcopically directed punch biopsies were taken from each CIN case concurrently with high-risk HPV testing by polymerase chain reaction (PCR). Results: The mean age of the patients was 38.69 (SD ±7.76) years. CIN 1 was diagnosed in 57% of participants, while 24% had CIN II and 19% had CIN III lesions. High-risk HPV was present in 52 patients. HPV 16 was the most common identified in 28 (53.84%) and HPV 18 was the second most common with 20 (38.46%) either singly or in combination with other high-risk subtypes. The other HPV strains, HPV 31, 33, 35, 52, 56 and 58, were also detected either as mono or co-infections. Out of the 52 HPV positive cases, 29 (55.8%) had mono infection and 23 (44.2%) had co-infection with several subtypes. The highest incidence (50%) of oncogenic HPV infections was present among women aged 35 - 45 years. Risk factors associated with HPV positive cases were high parity (P 0.05), early age at marriage (P = 0.754) and early age of first child. Conclusion: This study identified a high prevalence of HPV 16 and 18 genotypes. HPV vaccination with the current 9-valent HPV vaccine, which contains HPV types 6, 11, 16, 18, 31, 33, 45, 52, and 58. Will be an effective public health measure to eradicate cervical cancer in Bangladesh.

Personalized precision radiotherapy and its evolving role for human papillomavirus-positive oropharyngeal cancer

Human papilloma virus(HPV)-associated oropharyngeal cancer(OPC)is a unique entity with increased respon-siveness to treatment and excellent oncologic outcomes.The purpose of this narrative review is to highlight how an improved prognosis for HPV(+)tumors and an ever-increasing understanding of the risk factors,risk stratification,and areas of potential spread are shaping management options.Additionally,we aim to detail how advances in treatment technology on both the surgical and radiation fronts are facilitating the delivery of increas-ingly personalized and precise treatments.This review will describe key aspects of recent and currently-ongoing trials investigating the de-escalation and individualization of treatment in this patient cohort,and how they are building a foundation for distinct treatment paradigms for HPV(+)tumors.Further studies into the integration of biomarker-guided treatments combined with clinical trial enrollment will help ensure a future of personalized treatments and improved outcomes,both in terms of oncologic outcomes and toxicity,for patients with HPV(+)OPC.

New cheaper human papilloma virus mass screening strategy reduces cervical cancer incidence in Changsha city:A clinical trial

BACKGROUND Cervical cancer is the second leading cause of death in women worldwide,second only to breast cancer.Around 80%of women have been infected with human papillomavirus(HPV)in their lifetime.Early screening and treatment are effective means of preventing cervical cancer,but due to economic reasons,many parts of the world do not have free screening programs to protect women’s health.AIM To increase HPV cervical cancer screening in Changsha and reduce the incidence of cervical cancer.METHODS Cervical cancer screening included gynecological examination,vaginal secretion examination and HPV high-risk typing testing.Cervical cytology examination(ThinPrep cytology test)was performed for individuals who test positive for HPV types other than 16 and 18.Vaginal colposcopy examination was performed for HPV16 and 18 positive individuals,as well as for those who were positive for ThinPrep cytology test.If the results of vaginal colposcopy examination were abnormal,histopathological examination was performed.We conducted a cost-benefit analysis after 4 years.RESULTS From 2019 to 2022,523437 women aged 35-64 years in Changsha city were screened and 73313 were positive,with a 14%positive rate.The detection rate of precancerous lesions of cervical cancer was 0.6%and the detection rate of cervical cancer was 0.037%.Among 311212 patients who underwent two cancers examinations,the incidence rate was reduced by more than half in the second examination.The average screening cost per woman was 120 RMB.The average cost of detecting early cases was 10619 RMB,with an early detection cost coefficient of 0.083.CONCLUSION Our screening strategy was effective and cost-effective,making it valuable for early diagnosis and treatment of cervical cancer.It is worth promoting in economically limited areas.

Expression and clinical significance of high risk human papillomavirus and invasive gene in cervical carcinoma

Objective:To study the expression of E6 and E7 mRNA in high-risk human papillomavirus(HPV) HPV-18 and the relationship between the expression of invasive gene and cervical carcinoma.Methods:A total of 119 patients with cervical cancer,cervical erosion and cervical HPV infection who were diagnosed in our hospital were selected and randomly divided into two groups:cervical cancer group(n= 58) and non-cancerous group(n= 61).Another 60 patients with uterine leiomyoma were selected as normal control group.Detection of HPV18 E6,E7 mRNA expression and invasion,migration,proliferation inhibition genes,epithelial mesenchymal transition genes and proliferation related protein content.Results:The relative expression of E6 and E7 HPV-18 in cervical cancer group was significant higher than that in non-cancerous group and control group(mRNA)(P<0.05).The content of TRAF6 and c-FLIP in invasive cervical cancer group was significantly higher than that in non-cancerous group and control group(P<0.05).The mR NA content of CD44v6 and MMP-9 in cervical cancer group was significantly higher than that in non-cancerous group and control group(P<0.05).The content of DEC-1,IKK16,MBP-1 in cervical cancer group was significant lower than that in non-cancerous group and control group(P<0.05).The mR NA content of beta-catenin and Vimentin in cervical cancer group was significantly lower than that in non cancerous group and control group(P<0.05).The proliferation related protein E2F1 of cervical cancer group was significantly lower than that of non-cancerous group and control group,Bmi-1 content was significantly higher than non-cancerous group and control group(P<0.05).Conclusions:The expression of the detection of cervical cancer in high-risk human papilloma virus HPV-18 E6 and E7 mRNA,and the invasion,migration,proliferation inhibition gene,epithelial mesenchymal transition and proliferation related gene protein content,HPV expression rate of mR NA increased with the development of cervical cancer,the expression is also enhanced.The expression has a certain correlation between the level and development of cervical cancer.Through the above indicators,the development of cervical cancer monitoring and treatment to provide important clinical guidance.

Human papillomavirus in esophageal squamous cell carcinoma in Colombia and Chile

AIM： To examine the presence of human papillomavirus （HPV） in esophageal squamous cell carcinoma （ESCC） specimens collected from Colombia and Chile located in the northern and southern ends of the continent, respectively.METHODS： We examined 47 and 26 formalin-fixed and paraffin-embedded ESCC specimens from Colombia and Chile, respectively. HPV was detected using GP5＋/GP6＋ primer pair for PCR, and confirmed by Southern blot analysis. Sequencing analysis of L1 region fragment was used to identify HPV genotype. In addition, P16^INK4A protein immunostaining of all the specimens was conducted.RESULTS： HPV was detected in 21 ESCC specimens （29%）. Sequencing analysis of L1 region fragment identified HPV-16 genome in 6 Colombian cases （13%） and in 5 Chilean cases （19%）. HPV-18 was detected in i0 cases （21%） in Colombia but not in any Chilean case. Since Chilean ESCC cases had a higher prevalence of HPV-16 （without statistical significance）, but a significantly lower prevalence of HPV-18 than in Colombian cases （P = 0.011） even though the two countries have similar ESCC incidence rates, the frequency of HPV-related ESCC may not be strongly affected by risk factors affecting the incidence of ESCC. HPV-16 genome was more frequently detected in p16 positive carcinomas, although the difference was not statistically significant. HPV-18 detection rate did not show any association with p16 expression. Well-differentiated tumors tended to have either HPV-16 or HPV-18 but the association was not statistically significant. HPV genotypes other than HPV-16 or 18 were not detected in either country.CONCLUSION： HPV-16 and HPV-18 genotypes can be found in ESCC specimens collected from two South American countries. Further studies on the relationship between HPV-16 presence and p16 expression in ESCC would aid understanding of the mechanism underlying the presence of HPV in ESCC.

Accuracy of triage strategies for human papillomavirus DNA-positive women in low-resource settings:A cross-sectional study in China

Objective： CareHPV is a human papillomavirus （HPV） DNA test for low-resource settings （LRS）. This study assesses optimum triage strategies for careHPV-positive women in LRS. Methods： A total of 2,530 Chinese women were concurrently screened for cervical cancer with visual inspection with acetic acid （VIA）, liquid-based cytology and HPV testing by physician- and self-collected careHPV, and physician-collected Hybrid Capture 2 （HC2）. Screen-positive women were referred to colposcopy with biopsy and endocervical curettage as necessary. HPV-positivity was defined as _〉1.0 relative light units/cutoff （RLU/CO） for both careHPV and HC2. Primary physician-HC2, physician-careHPV and self-careHPV and in sequential screening with cytology, VIA, or increased HPV test-positivity performance, stratified by age, were assessed for cervical intraepithelial neoplasia （CIN） grade 2/3 or worse （CIN2/3＋） detection. Results： The sensitivities and specificities of primary HPV testing for CIN2＋ were： 83.8%, 88.1% for physician- careHPV; 72. 1%, 88.2% for self-careHPV; and 97.1%, 86.0% for HC2. Physician-careHPV test-positive women with VIA triage had a sensitivity of 30.9% for CIN2＋ versus 80.9% with cytology triage. Self-careHPV test- positive women with VIA triage was 26.5% versus 66.2 % with cytology triage. The sensitivity of HC2 test-positive women with VIA triage was 38.2 % versus 92.6% with cytology triage. The sensitivity of physician-careHPV testing for CIN2＋ decreased from 83.8% at _〉1.0 RLU/CO to 72.1% at _〉10.00 RLU/CO, while the sensitivity of self- careHPV testing decreased from 72.1% at _〉1.0 RLU/CO to 32.4% at _〉10.00 RLU/CO; similar trends were seen with age-stratification. Conclusions： VIA and cytology triage improved specificity for CIN2/3 than no triage. Sensitivity with VIA triage was unsuitable for a mass-screening program. VIA provider training might improve this strategy. Cytology triage could be feasible where a high-quality cytology program exists. Triage of HPV test-positive women by increased test positivity cutoff adds another LRS triage option.

Identification of human papillomavirus in esophageal squamous papillomas

AIM： To investigate the presence of human papillomavirus （HPV） in esophageal squamous papilloma （ESP） and determine p16, p53 and Ki67 expression in a Mexican cohort. METHODS： Nineteen cases diagnosed as ESP, corresponding to 18 patients were reviewed; nineteen cases of normal esophageal mucosa were used as negative controls. HPV detection was performed by ,amplified chromogenic in situ hybridization （ACISH） using a wide spectrum-cocktail probe and PCR. RESULTS： The average age at presentation was 46.3 years （range 28-72 years）. Patients included four （22.22%） males and 14 （77.77%） females. The most frequent location was upper third （11 cases）, followed by middle third （3 cases） and unknown site （5 cases）. Immunohistochemistry （IHC） revealed basal and focal p53 expression in 17 cases （89%）; p16 was expressed in eight cases （42.10%） and the Ki67 index ranged from 10% to 30%. HPV was detected in 14 out of 16 cases （87.5%） by ACISH： Twelve showed diffuse nuclear patterns and two showed granular patterns. HPV DNA was identified by PCR in 12 out of 14 cases （85.7%）. Low-risk HPV types were detected in the most of the cases. CONCLUSION： This study provides identification of HPV infection in almost 80% of ESP using either ACISH or PCR; overall, all of these lesions show low expression of cell-cycle markers. We suggest ACISH as an alternative diagnostic tool for HPV detection in ESR .

Association between Helicobacterpylori, Epstein-Barr virus, human papillomavirus and gastric adenocarcinomas

AIM TO correlate Helicobacter pylori （H. pylorli）, EpsteinBarr virus （EBV） and human papillomavirus （HPV） with gastric cancer （GC） cases in Para State, Brazil.METHODS Tissue samples were obtained from 302 gastric adenocarcinomas. A rapid urease test was used to detect the presence of H. pylori, and the presence of the cagA gene in the HP-positive samples was confirmed by PCR. An RNA in situ hybridization test designed to complement Eberl RNA was used to detect the presence of EBV in the samples, and the L1 region of HPV was detected using nested PCR. Positive HPV samples were genotyped and analyzed for E6 and E7 viral gene expression. Infections were also correlated with the clinical and pathological characteristics of the patients.RESULTS The majority of the 302 samples analyzed were obtained from men （65%） aged 55 years or older （67%） and were classified as the intestinal subtype （55%）. All three pathogens were found in the samples analyzed in the present study （H. pylori： 87%, EBV： 20%, HPV： 3%）. Overall, 78% of the H. pylori-positive （H. pylorl＋） samples were cagA＋ （H. pylori-cagA＋）, and there was an association between the cytotoxic product of this gene and EBV. Coinfections of H. pylori-cagA＋ and EBV were correlated with the most advanced tumor stages. Although only 20% of the tumors were positive for EBV, infection with this virus was associated with distant metastasis. Only the HPV 16 and 18 strains were found in the samples, although no expression of the E6 and E7 oncoproteins was detected. The fundus of the stomach was the region least affected by the pathogens.CONCLUSION HPV was not involved in gastric tumorigenesis. Prophy- lactic and therapeutic measures against H. pylori and EBV may prevent the development of GC, especially the more aggressive forms.

Integration of human papillomavirus 18 DNA in esophageal carcinoma 109 cells

AIM:To detect human papillomavirus(HPV) DNA in esophageal carcinoma(EC) 109 cells and investigate the relationship between HPV and EC.METHODS:Genomic DNA and total RNA from EC109 cells were isolated.HPV DNA was detected by polymerase chain reaction(PCR) with the general primer sets of My09/11 and GP5 +/6 + for the HPV L1 gene and type-specific primer sets for HPV18 E6 and HPV18 E6-E7.Reverse transcription(RT) of mRNA isolated from EC109 cells was performed to produce a cDNA.And then a PCR-based protocol for the amplification of papillomavirus oncogene transcripts was used to analyze HPV18 DNA and integrated transcripts of HPV18 in the chromosomes of EC109 cells.The final nested PCR products were cloned into a pMD-18T vector and sequenced to analyze the chromosomal location of HPV integration.RESULTS:HPV18 DNA was detected in EC109 cells by PCR using the general primer sets of My09/11 and GP5 +/6 + for HPV L1 and the type-specif ic primer sets for HPV18 E6 and E6-E7 to generate products of 450 bp,150 bp,335 bp and 944 bp,respectively.Approximately 600 bp of integrated HPV18-specific transcript was identified.The final nested PCR product of integrated HPV18 DNA was cloned into a pMD-18T vector and sequenced to analyze the chromosomal location of HPV integration.Sequence alignment showed that the HPV18 sequence from EC109 cells was identical to that of the encoded early protein E7-E1 of the standard HPV18 strain X05015,and another partial gene sequence was identical to a partial sequence of human chromosome 8.CONCLUSION:Integration of the HPV genome into the host cell chromosome suggests that persistent HPV infection is vital for malignant cell transformation and carcinogenesis.

Comparison of Human Papillomavirus Detection and Genotyping with Four Different Prime Sets by PCR-Sequencing

Objective To assess and compare the Human Papillomavirus（HPV） detection efficiency and the potential clinical utility of PCR sequencing‐based technology.Methods Four HPV consensus primer sets（GP5＋/6＋,MGP,MY09/11,and PGMY09/11） were used in order to amplify a broad spectrum of HPV types for HPV infection in 325 cervical samples and the PCR products were sequenced afterwards for the HPV genotyping.Results The HPV‐positive rate was 75.4%,of which 35.5% harbored more than one HPV genotype.A total of 36 different genotypes was found,with HPV 16（24.1%） being the most prevalent,followed by HPV 58（13.3%） and HPV 52（9.6%）.There were substantial to almost perfect agreements between different primer sets regarding HPV detection efficiency,with the kappa value varying from 0.751 to 0.925,MGP,and PGMY09/11 were the most effective in detecting multiple infections（P0.001）.With each of the primer sets,a board range of HPV types could be identified,though there were several differences for a few genotypes.Conclusion The substantial agreement between PCR‐sequencing and HC2 for the detection of high‐risk HPV（kappa=0.761） indicated that PCR‐sequencing is also suitable for routine HPV screening.

Urine versus brushed samples in human papillomavirus screening： study in both genders

Aim： To investigate whether urine is a good medium for screening and whether there is a correlation between the amount of extracted DNA and human papillomavirus （HPV）-positivity. Methods： In the present study, 30 first-voided urine （FVU） specimens and 20 urethroglandular swabs using cervex-brushes from male partners of HPV-positive patients, and 31 FVU specimens and 100 liquid-based cervix cytology leftovers sampled with cervix-brushes from HPV-positive women were examined for the presence of β-globin. Oncogenic HPV were detected using type-specific PCR. Results： β-globin was found in all the brushed samples, whereas it was found in only 68.9% of the FVU specimens. HPV-PCR was positive in 60.0% of the male brushes, in 29% of the female brushes and in 0% of the male FVU specimens. DNA concentration was, respectively, 0.9998 ng/μL, 37.0598 ng/μL and 0.0207 ng/μL. Conclusion： Urine is not a good tool for HPV detection, probably because the low DNA concentration reflects a low amount of collected cells. β-globin is measurable in FVU by real time quantitative PCR, but the DNA concentration is lower compared to brush sampling for both genders. β-globin-positivity of urethral and cervical swabs is 100%, showing a higher mean concentration of DNA, leading to a higher detection rate of HPV. This is the first article linking DNA- concentration to the presence of HPV.