PRELIMINARY STUDY OF A NOVEL HUMAN PAPILLOMAVIRUS TYPE 16 L1/E6-E7 CHIMERIC RECOMBINANT DNA VACCINE

Objective Preparations of HPV16 L1/E6 and L1/E7 prophylactic and therapeutic DNA vaccines. Methods The nucleotides within HPV16 E6 and E7 genes, which are responsible for viral transforming activity, were mutated by mage primer site-directed mutagenesis method. The correctly mutated E6 and E7 fragments were separately cloned into an eukaryotic expression vector pVAX1, together with HPV16 L1 gene, generating chimeric recombinants plasmids 1MpVAX1-L1E6, 2MpVAX1-L1E6, 1MpVAX1-L1E7, 2MpVAX1-L1E7 and 3MpVAX1-L1E7. CHO cells were transiently transfected with the individual DNA vaccines by calcium phosphate method. Target protein expressions in the extracts of the transfected cell lines were measured by ELISA and immunohistochemistry, with HPV16 L1 and E6 specific monoclonal antibodies. Results ELISA assays showed the P/N ratios in the cell extracts transfected with L1E6 and L1E7 plasmids were more than 2.1. Immunohistochemistry revealed brownish precipitant signal in cytoplasm and nuclei of the transfected cells. Conclusion Successful constructions of prophylactic and therapeutic DNA vaccine plasmids lay solid foundation for future animal experiment and clinical trial.

SPECIFIC UP-REGULATION OF DNA POLYMERASE BY HUMAN PAPILLOMAVIRUS 16

Obje, etive To analyze how the infection of human papillomavirus 16 （ HPV16 ） affects expression of DNA poly- merase β（DNA polB） with the aim of probing the mechanism of over-expression of DNA polB in human cancers. Methods Four fragments of human DNA polB promoter were amplified and constructed into luciferase reporter vec- tor pGL-Basic, generating pGL-BP, pGL-BMH, pGL-BMS, and pGL-BAT constructs respectively, and co-transfected with HPV16 or HPV6 into Hep2 cells. Luciferase activity was assayed 48 hours after transfectiorL Semi-quantitative RT- PCR was used to measure mRNA expression of endogenous DNA polB. Immunohistochemistry and in situ hybridization were used to analyze DNA polB expression and HPV16 or HPV6 infection in 38 cases of cervical lesions respectively. Results With co-transfection of HPV16 and DNA polB promoter-driving reporters into Hep2 cells, pGL-BP re- porter in full-length DNA polB promoter presented markedly elevated luciferase activities （ P 〈 0.05 ）. However, the other three mutant reporters： pGL-BMH, pGL-BMS, and pGL-BAT, generated no reporting activities in the presence of HPV16 （P 〉0.05 ）. On the contrary, all of polB promoter reporters were little stimulated in co-transfection of HPV6 （P 〉0.05 ）. The transfection of HPV16 could enhance the endogenous polB mRNA expression compared with that of HPV6 （3.42 vs. 0.80, P 〈0.05）. The DNA polB expression was found in 8 of 10 HPV16-positive cervical intraepi- thelial neoplasia grade Ⅲ（ CIN Ⅲ） cases, while was only found in 3 of 11 HPV6-positive condyloma accuminatum cases, but was negative in all chronic cervicitis cases. The correlation of DNA polB expression with HPV16 infection in cervical lesions was significant （ P 〈0.05 ）. Conclusions HPV16 is able to specifically stimulate the expression of DNA polB in human epithelial cells through interaction with the core upstream regulatory sequences of DNA polB promoter. Over-expression of DNA polB might be an explanation for the molecular mechanism underlying HPV-related human cancers.

Detection of Human Papillomavirus Type 16 DNA in Cervical Carcinomas

Accurate typing of the different human papillomavirus types is csscntial in view of the differ-ent pathological potential of the common virus types of human papillomavirus （HPV） present in thecervix. We have developed hybridization, washing and autoradiography conditions that minimize thecross-hybridization among different specific types of HPV so as to allow clear - cut type assignmentthrough practical dot blot hybridization technique using nylon membrane and 35S - labeled HPV - 16DNA probe. Under these conditions seventeen of thirty （56.7%） of squamous cell carcinomas of thecervix uteri obtained from Tianjin women were detected in the presence of HPV - 16 DNA.

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.

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 the Tellgenplex HPV DNA test with the PCR-reverse dot blot assay for human papillomavirus genotyping

Objective: To access the performance of the Tellgenplex human papillomavirus(HPV) DNA test compared to the polymerase chain reaction-reverse dot blot(PCR-RDB) assay for the HPV genotyping.Methods: Sixty cervical swab samples were genotyped by the Tellgenplex HPV DNA test and the PCR-RDB assay.The Tellgenplex HPV DNA test and the PCR-RDB assay can detect 26 and 23 HPV genotypes, respectively.Each sample showed discrepancy was genotyped using sequencing.Results: The percent agreement between the two tests ranged from 83.3% to 100.0% according to different genotype.This showed perfect agreement(>0.81) for high-risk HPV genotypes(35, 39, 45, 53, 56, 59, 66, 68, and 82), substantial agreement(>0.65) for high-risk HPV genotypes(16, 18, 33, 52, and 58) and low-risk HPV genotype 43 between the two assays by the kappa analysis.The positive rates of the two assays for frequent HPV genotypes(16, 35, 39, 45, 52, 53, 58, 59, 66, and 82) were not statistically different, but the PCR-RDB assay showed higher positive rates than the Tellgenplex HPV DNA test for HPV genotypes 81(P<0.05).As for more than 10 positive results by the Tellgenplex HPV DNA test and/or the PCR-RDB assay, the PCR-RDB assay showed higher relative sensitivity and specificity than the Tellgenplex HPV DNA test for the three HPV genotypes(16, 52, and 81).All HPV genotypes that can be detected by only the Tellgenplex HPV DNA test(HPV genotypes 44 and 55) were confirmed by sequencing.Conclusions: In conclusion, our results demonstrated that the PCR-RDB assay which can detect more multiple HPV genotypes in each specimen shows higher relative sensitivity and specificity than the Tellgenplex HPV DNA test, which makes it a better option for routine clinical use.

Human papillomavirus DNA and P16～(INK4A) expression in concurrent esophageal and gastric cardia cancers

AIM:To investigate the relationship between human papillomavirus (HPV) infection and concurrent esophagus and gastric cardia cancer from the same patient (CC) and examine the significance of P16 INK4A protein expression.METHODS:Polymerase chain reaction was used to detect the presence of HPV type16 (HPV16).The expression of P16 INK4A protein was detected using immunohistochemistry.RESULTS:Among the CC specimens,HPV16-DNA was found in eight cases of esophageal squamous cell carcinoma (ESCC) and five cases of gastric cardia adenocarcinoma (GCA),respectively (47% vs 29%),and two of both ESCC and GCA.P16 INK4A was highly expressed in both ESCC and GCA.In the HPV-associated positive CC,higher P16 INK4A expression was observed in the GCA than in the ESCC (75% vs 25%,P < 0.05).CONCLUSION:HPV16 as a correlated risk factor may play an important role in the development of ESCC and GCA.P16 INK4A may be a screening index in the HPVassociated carcinoma of gastric cardia.

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.

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.

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.

The relationship between DNA fragmentation and the intensity of morphologically abnormal human spermatozoa

Objective:To determine the relationship between teratozoospermia and sperm DNA fragmentation(SDF)in the human ejaculate.Methods:This retrospective study included 100 normozoospermic men as a control cohort(abnormal forms>14%),210 patients with a high level of abnormal forms(≤4%)and 65 patients presenting with a moderate level of abnormal forms(>4%to≤14%)based on the World Health Organization definitions.Sperm morphology was assessed using bright field microscopy.Sperm DNA fragmentation was assessed using the sperm chromatin dispersion assay.Non-parametric analyses were conducted to examine the relationship between abnormal sperm morphology and sperm DNA fragmentation;receiver operating characteristic(ROC)analyses were conducted to assess sensitivity and specificity of this relationship.Results:A correlation analysis revealed that the higher the proportion of abnormal spermatozoa in the ejaculate,the higher the level of SDF(Spearman's Rho=-0.230;P<0.001).Significant differences in the proportion of SDF were found when all cohorts were compared(P<0.001);these significant differences were also retained when the different cohorts were compared pairwise.ROC analysis showed a moderate but significant predictive value for SDF to differentiate patients with different levels of teratozoospemia.Conclusions:Although analysis of a more continuous range of values for teratozoospermia would help further clarify any causal relationship with SDF,there is clearly a synergistic or coincident affiliation between these variables that needs to be acknowledged by the clinician when interpreting the spermiogram.

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.

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.

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.

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.

CONSTRUCTION AND IMMUNOGENICITY OF HUMAN PAPILLOMAVIRUS TYPE 6B L1 RECOMBINANT PLASMID

To construct a DNA vaccine as a prophylactic model to prevent condyloma acuminatum and detect its immu-nogenicity in mice. Methods The major capsid protein (L1) gene of human papillomavirus (HPV) 6b was inserted into an eukaryotic ex-pression plasmid (pcDNA3.1). The recombinant plasmid was transfected into COS-7 cells. Western blot were performed to detect whether L1 protein can be expressed in eukaryotic cells. Eighteen female BALB/c mice were tested for immunoge-nicity study. Results The recombinant plasmid (pcDNA3.1-HPV6bL1) was verified as HPV6b L1 gene by sequencing. Western blot showed specific strip. Anti-L1 protein antibodies could be detected in the mice’s sera inoculated with pcDNA3.1-HPV6bL1. Similarly, IL-4, IL-2, and IFN-γ were increased in the same mice. Conclusion HPV6b L1 recombinant plasmid was constructed successfully which had immunogenicity for BALB/c mice. It provided experimental evidence for the research of DNA vaccine of condyloma acuminata..-

Exploring Genome-wide DNA Methylation Profiles Altered in Kashin-Beck Disease Using Infinium Human Methylation 450 Bead Chips

To understand how differentially methylated genes（DMGs）might affect the pathogenesis of Kashin-Beck disease（KBD）.Genome-wide methylation profiling of whole blood from 12matched KBD and controls pairs was performed using a high-resolution Infinium 450 K methylation array.In total,97 CpG sites were differentially

Hexabromocyclododecane-induced Genotoxicity in Cultured Human Breast Cells through DNA Damage

To investigate the genotoxicity and reveal the potential toxicological mechanisms of Hexabromocyclododecane （HBCD）, human breast cells HBL-100 were exposed to a sequence of HBCD concentrations （0, 5, 10, and 50 mg/L） for 24 h. With a series of zymology and molecular biology methods, we found that HBCD induced dose-dependent oxidative stress on HBL-100 DNA. As revealed in q RT-PCR, activated prognostic factor ATM down-regulated tumor suppressor gene BRCA1 and prompted DNA repair genes h OGG1 and h MTH1 expression in lower concentrations of HBCD （〈 10 mg/L）. However, DNA repair were inhibited as well as cell proliferation rate by higher concentrations of HBCD （50 mg/L）. The results inferred that the genotoxicity of HBCD was dose-dependent and related to DNA repair pathway.