Association of Human Papilloma Virus Infection and Oral Squamous Cell Carcinoma in Bangladesh

Oral squamous cell carcinoma is the sixth most common malignancy worldwide. In Bangladesh, it comprises 20% of the whole body malignancies. Several studies found that 15% to 25% of oropharyngeal cancer cases are associated with human papilloma virus (HPV). This study is done to find the association of human papilloma virus subtypes, particularly HPV type 16 and HPV type 18, with the oral squamous cell carcinoma in Bangladeshi patients. In total, 34 diagnosed patients of oral squamous cell carcinoma were included in the study. Extracted DNA from the cancerous tissues was checked for PCR reaction to detect the subtypes of human papilloma virus. Data of the present study suggest that oral squamous cell carcinoma are almost absent in Bangladeshi patients with human papilloma virus, particularly HPV 16 and 18.Key words: Human papilloma virus (HPV), HPV type 16, HPV type 18, Oral cancer, Polymerase chain reaction, Bangladesh     

人乳头瘤病毒11型L1 DNA疫苗质粒的构建和鉴定

目的:构建人乳头瘤病毒11型(HPV11)L1DNA疫苗质粒。方法:从尖锐湿疣病变组织中提取DNA制备模板,采用聚合酶链反应(PCR)技术扩增HPV11-L1基因,将L1基因经双酶切后克隆到pcDNA3 1(+)中,测序鉴定。结果:从病变组织中扩增出了全长HPV11-L1基因,并正向插入表达载体pcDNA3 1(+)中,转化JM109扩增后经酶切和测序鉴定,证实克隆成功。结论:HPV11-L1DNA疫苗质粒的构建为今后进行防治尖锐湿疣的动物实验和临床实验奠定了基础。     

人乳头瘤病毒11型E7与共刺激分子CD80嵌合DNA疫苗质粒的构建及鉴定

目的:构建人乳头瘤病毒11型(HPV ll)E7与共刺激分子CDS0嵌合DNA疫苗质粒。方法:从尖锐湿疣病变组织中提取DNA制备模板,采用聚合酶链反应(PCR)技术扩增HPV ll-E7基因,经双酶切后定向克隆到pcDNA3.1(+)中,获得pcDNA3.1(+)/HPV llE7,测序鉴定后,再用PCR方法扩增去除终止密码子的E7基因片段,按上述方法插入质粒pcD-NA3.1(+)/CD80中CD80上游,构建pcDNA3.1(+)/HPV llE7-CD80融合基因真核表达质粒。结果:去除终止子的E7基因片段成功正向插入CD80上游,双向测序分析显示序列无误,pcDNA3.1(+)/HPV llE7-CD80嵌合真核表达质粒构建成功。结论:HPV ll-E7/CD80嵌合DNA疫苗质粒构建成功,为研究该疫苗在尖锐湿疣防治中的作用奠定了基础。     

Intramuscular injection of plasmid DNA encoding cottontail rabbit papillomavirus E1, E2, E6 and E7 induces T cell-mediated but not humoral immune responses in rabbits

To test the efficacy of genetic vaccination against papillomavirus infection, plasmid DNA encoding cottontail rabbit papillomavirus (CRPV) E1, E2, E6, E7 or without insert were intramuscularly injected into five groups of rabbits. Peripheral blood mononuclear cells (PBMCs) showed specific proliferation upon in vitro stimulation with E1, E2, E6 or E7 proteins in a majority of vaccinated rabbits but Western blot analysis did not detect antibodies specific for these viral proteins in rabbit serum. All rabbits grew papillomas after virus challenge and none of the rabbits showed systemic papilloma regression. These observations showed that intramuscular injection of plasmid DNA encoding CRPV E1, E2, E6 or E7 induced CD4+ T cell-mediated but not humoral immune responses, and did not result in the protection of rabbits from virus infection.     

Insulin action on expression of novel adipose genes in healthy and type 2 diabetic subjects

OBJECTIVE: Adipose tissue secretes several molecules that may participate in metabolic cross-talk to other insulin-sensitive tissues. Thus, adipose tissue is a key endocrine organ that regulates insulin sensitivity in other peripheral insulin target tissues. We have studied the expression and acute insulin regulation of novel genes expressed in adipose tissue that are implicated in the control of whole body insulin sensitivity. RESEARCH METHODS AND PROCEDURES: Expression of adiponectin, c-Cbl-associated protein (CAP), 11-beta hydroxysteroid dehydrogenase type 1 (11beta-HSD-1), and sterol regulatory element binding protein (SREBP)-1c was determined in subcutaneous adipose tissue from type 2 diabetic and age- and BMI-matched healthy men by real-time polymerase chain reaction analysis. RESULTS: Expression of adiponectin, CAP, 11beta-HSD-1, and SREBP-1c was similar between healthy and type 2 diabetic subjects. Insulin infusion for 3 hours did not affect expression of CAP, 11beta-HSD-1, or adiponectin mRNA in either group. However, insulin infusion increased SREBP-1c expression by 80% in healthy, but not in type 2 diabetic, subjects. DISCUSSION: Our results provide evidence that insulin action on SREBP-1c is dysregulated in adipose tissue from type 2 diabetic subjects. Impaired insulin regulation on gene expression of select targets in adipose tissue may contribute to the pathogenesis of type 2 diabetes.     

陕西地区妇女乳腺癌组织中58型人乳头瘤病毒感染分析

目的：检测乳腺癌组织中58型人乳头瘤病毒E7基因，探讨人乳腺癌组织中是否存在HPV58感染。方法采用聚合酶链反应技术（PCR），结合人乳头瘤病毒（HPV）型特异性引物，检测了141例乳腺癌组织和55例非癌乳腺病变组织中的HPV58 E7基因。结果141例乳腺癌标本PCR结果显示，HPV58 E7基因检出率为12．06％（17/141），55例非癌乳腺病变组织中HPV58 E7基因检出率为1．82％（1/55），两者存在显著性差异（χ2＝4．973，P＝0．026）。结论 HPV58在乳腺癌组织中有相当高的感染率，提示可能与乳腺癌的发生有关。     

人乳头瘤病毒16型感染与食管癌发生相关性的Meta分析

[目的]探讨人乳头瘤病毒(Human Papillomavirus,HPV)感染与我国与食管癌发生的病因学关系。[方法]在国内万方网,检索有关HPV与食管癌相关的研究文献,研究纳入标准是:以聚合酶链反应或原位杂交为检测方法检测HPVDNA,并且研究病例既有食管癌组织又有正常对照组织。将所得数据进行Meta分析。[结果]入选12篇文献进行HPV16型特异性检测,其结果显示,食管癌组织标本中HPV16占38.6,正常组织标本中HPV16占15.9,HPV16感染率与正常对照组织相比差异有统计学意义(P﹤0.05)。[结论]高危型HPV16感染可能是食管癌发生的重要病原因子。     

Therapeutic vaccination of rabbits with a ubiquitin-fused papillomavirus E1, E2, E6 and E7 DNA vaccine

Previously, we showed that intracutaneous vaccination of rabbits with DNA vectors encoding ubiquitin-fused versions of the cottontail rabbit papillomavirus (CRPV) early proteins E1, E2, E6 and E7 protected against subsequent challenge with CRPV. Here, we tested the immunotherapeutic activity of a vaccine composed of the four CRPV DNA vectors (designated UbE1267) in rabbits. The results show that the UbE1267 DNA vaccine, relative to empty vector DNA, virtually eliminated papilloma growth in rabbits with subclinical infection and greatly reduced papilloma volumes in rabbits bearing papillomas at the time of vaccination. These results in a physiologically relevant animal model of high-risk human papillomavirus (HPV) infection indicate that DNA vaccines targeting the early papillomavirus proteins may have a role in the treatment of HPV-associated lesions in humans.     

Human papillomaviruses in Amerindian women from Brazilian Amazonia

We evaluated the prevalence of human papillomavirus (HPV) infection in Amerindian women from a tribe in Brazilian Amazonia. Demographic data, pap smears and cervical samples for HPV DNA detection by polymerase chain reaction (PCR) were obtained for women aged above 10 years old. In total, 79 (85.9%) out of 92 eligible women who lived there were interviewed; all women already had engaged in sexual activity. Seventy-eight and 49 women allowed collection of pap smears and PCR samples, respectively. Cytological signs of HPV infection were observed in 11 patients; 6 of these were probed for HPV infection and 1 shown to be HPV 16. Overall prevalence of HPV infection detected by PCR was 14.3%. Three patients presented high-risk HPV DNA types:two HPV 16 and one co-infection of HPV 16 and 58. Cervical infection by oncogenic HPV types occurs in Amerindian women and cervical cancer screening should be a priority in this setting.     

HUMAN PAPILLOMAVIRUS HPV-16 DNA AS AN EPITHELIOTROPIC VIRUS THAT INDUCES HYPERPROLIFERATION IN SQUAMOUS PENILE TISSUE

The prevalence of human papillomavirus HPV-16DNA sequences in 57 penile carcinoma biopsies was examined using the polymerase chain reaction (PCR) with type specific internal probes, employing HPV consensus primers from the L1 region. The cases comprised 39 typical squamous cell carcinoma and 18 specimens with different subtype. PCR products were analyzed and HPV-16DNA was detected in a high percentage of specimens. Thirty-eight biopsies were HPV-16DNA positive. This determination was correlated with cellular differentiation and growth pattern. Our data corroborates that squamous cell carcinoma was invariably associated with HPV-16DNA.     

人乳头瘤病毒(HPV)6/11、16/18型检测及结果分析的临床意义

目的用实时荧光定量PCR法检测低危型人乳头状瘤病毒HPV(如6/11型)和高危型人乳头状瘤病毒HPV(如16/18型),探讨其在宫颈癌防治方面的意义。方法采用FQ-PCR方法检测624例不同年龄组患者感染HPV6/11、HPV16/18型情况。结果624例标本中HPV6/11、HPV16/18检测的阳性率分别为26.3%,8.3%,低危型远高于高危型(P<0.01)。HPV6/11、HPV16/18阳性患者的年龄分布不同,年龄段的感染情况也不同,HPV6/11型感染主要集中在20～40岁,HPV16/18型感染者年龄均在41～50岁。结论荧光定量PCR检测低危型HPV和高危型HPV,方法简便准确,快速。     

Screening for autoantibodies in human subjects immunized with Pr-beta-HCG-TT.

Sera from 3 human volunteers immunised with Pr-β-HCG-TT vaccine were tested for possible reactivity with other human tissues. They were found to be negative for antinuclear, antimicrosomal antibodies and rheumatoid factor. The sera from these subjects did not give any reaction with human thyroid, pituitary, parathyroid, adrenal, testes and ovaries as examined by immunofluorescence techniques. They were also devoid of reaction with tissue substrates from baboons, mice and rabbits.     

基于实时荧光环介导等温扩增的人乳头瘤病毒58型基因检测

目的 建立基于实时荧光环介导等温扩增的人乳头瘤病58型基因检测体系，用于其快速检测和早期诊断。 方法 运用LAMP在线引物设计软件Primer Explorer V5, 针对人乳头瘤病毒58型的L1区域保守基因设计引物，在反应前加入荧光核酸染料SYBR Green Ⅰ，并对该反应体系中的dNTPs、Mg2+和内引物（FIP、BIP）浓度进行优化,优化结果通过“S”型扩增曲线判断，并对优化后的实时荧光环介导等温扩增体系进行特异性、灵敏度检测及临床标本检测。 结果 针对人乳头瘤病毒58型所建立的实时荧光环介导等温扩增检测体系具有良好的特异性及灵敏度，且与临床实验室检测结果（qPCR方法）的符合率达100%，该检测方法的灵敏度可达10拷贝/μL。 结论 本研究建立了人乳头瘤病毒58型的实时荧光环介导等温扩增检测体系，它是一种耗时少、操作简便、特异性强、灵敏度高的方法，适用于HPV临床样本的快速检测。     

Human papillomaviruses and cervical cancer in Bangkok. I. Risk factors for invasive cervical carcinomas with human papillomavirus types 16 and 18 DNA

Personal interviews, tests for antibodies to herpes simplex virus type 2, Treponema pallidum, and hepatitis B, tests for hepatitis B surface antigen (HBsAg), and polymerase chain reaction-based assays for human papillomavirus (HPV) DNA in cervical scrapings were obtained from 190 women with squamous cell and 42 women with adenomatous cervical carcinoma and from 291 hospitalized controls diagnosed in Bangkok, Thailand, between September 1991 and September 1993. Risk was strongly associated with oncogenic HPV types, with types 16 and 18 predominating in squamous and adenomatous lesions, respectively. The 126 cases with HPV-16 and the 42 cases with HPV-18 were compared with 250 controls with no evidence of any HPV. The risk of both viral tumor types increased with decreasing age at first intercourse in this predominantly monogamous population, which may be explained by more visits to prostitutes by the husbands of cases with early than late age at first intercourse. HPV-16 tumors were weakly associated with HBsAg carrier state and smoking. The risk of tumors of both viral types increased with parity and use of oral contraceptives but not with injectable progestogens. Factors that may predispose to persistent, oncogenic HPV-16 or -18 infection may include estrogens or progestins in the presence of estrogens, immunosuppression, and smoking, but other factors related to low socioeconomic status are also involved.     

复发性流产者妊娠早期E-钙粘素的表达及其与HPV感染的相关性

目的:探讨E-钙粘素(E-cadherin)在复发性流产者妊娠早期滋养层细胞中的表达及其与人乳头状瘤病毒(HPV)感染的相关性。方法:采用免疫组织化学与PCR法分别检测34例复发性流产者(观察组)与41例正常早孕女性(对照组)妊娠早期滋养层细胞中E-cadherin的表达水平及HPV-DNA。结果:观察组滋养细胞中E-cadherin的表达低于对照组(P<0.001),HPV感染率(29.4%)则高于对照组(11.8%,P=0.039);HPV感染阳性的标本中,观察组E-cadherin的阳性表达(20%)也低于对照组(100%,P=0.015)。结论:妊娠早期滋养层细胞E-cadherin的异常表达可能与复发性流产的发生有关,而HPV可能参与这一过程。     

乙型肝炎病毒前C区1896 G→A突变对母婴垂直传播的影响

目的:探讨乙型肝炎病毒(HBV)前C区1896 G→A点突变对母婴垂直传播的影响。方法:收集60例本院妇产科HBeAg阴性/HBV DNA阳性的孕妇血标本,同时收集这些孕妇的60例新生儿脐带血。应用PCR固相杂交法检测孕妇血清中HBV DNA前C区1896 G→A突变,荧光定量PCR(FQ-PCR)检测HBV孕妇血清和新生儿脐带血中HBV DNA载量。分析HBV前C区1896 G→A点突变及孕妇血清HBV DNA载量对母婴垂直传播率的影响。结果:60例孕妇血标本共检测到38例HBV前C区1896 G→A点突变(63.3%);突变组母婴垂直传播发生率为44.7%(17/38),未突变组母婴垂直传播发生率为40.9%(9/22),二组比较无统计学差异(χ2=0.0831,P>0.05)。孕妇血中HBV DNA高载量组(≥1×105 copies/ml)母婴垂直传播发生率为72.0%(18/25),低载量组(<1×105 copies/ml)母婴垂直传播发生率为22.9%(8/35),二组相比有显著性差异(χ2=14.3426,P<0.01)。结论:HBV前C区1896 G→A点突变未对母婴HBV垂直传播率造成影响,孕妇血清中HBV DNA载量升高增加了母婴HBV垂直传播的危险性。     

Therapeutic efficacy of vesicular stomatitis virus-based E6 vaccination in rabbits

Millions of people worldwide are currently infected with human papillomaviruses (HPVs). A therapeutic HPV vaccine would have widespread applicability because HPV-associated lesions are difficult to treat and may progress to carcinoma. We developed three attenuated VSV recombinants expressing the cottontail rabbit papillomavirus (CRPV) early protein E6 for use as vaccines. In cultured cells, two vectors expressed different levels of the E6 protein, and one expressed a ubiquitin-E6 fusion protein. All three were tested for therapeutic efficacy in the cottontail rabbit papillomavirus (CRPV)-rabbit model. Mock vaccination had no effect on papilloma growth. In contrast, inoculation with any of the VSV-E6 vaccines reduced the rate of papilloma growth to as little as 24% the rate in the controls. In five experiments, these effects were achieved after a single immunization. Furthermore, complete papilloma regression occurred in some rabbits observed for 4 months. A VSV-based papillomavirus E6 vaccine could have significant advantages over other therapeutic HPV vaccine candidates described to date.     

The sexually transmitted disease model for cervical cancer: incoherent epidemiologic findings and the role of misclassification of human papillomavirus infection.

The definition of cervical neoplasia as a sexually determined disease caused by some types of human papillomavirus has been widely accepted. Recent epidemiologic studies, however, have failed to identify a correlation between sexual activity and human papillomavirus infection. Moreover, sexual activity has also been shown to be independent of human papillomavirus infection in increasing cervical cancer risk. These incoherences are analyzed with respect to etiologic models for cervical neoplasia and by considering the role of misclassification of human papillomavirus infection in interpreting the relations assumed under these models. Even small levels of misclassification can considerably distort (1) the presumed prevalence of viral infection, (2) the association between sexual activity and human papillomavirus infection, and (3) the ability to control the relation between sexual activity and cancer by human papillomavirus infection. In field surveys, the presumed rates of human papillomavirus infection based on a DNA assay such as the filter in situ hybridization may be a gross overestimation of the true viral prevalence. Use of moderately misclassified human papillomavirus infection test results for effect estimation and covariate adjustment in data analysis may seriously distort the underlying relations. Consequently, considering these conditions, the apparent incoherence of recent epidemiologic findings should not be construed as evidence against the role of human papillomavirus in the etiology of cervical cancer or the validity of the sexually transmitted disease model.     

人乳头状瘤病毒负荷量与宫颈癌及宫颈癌前病变的相关研究

目的:探讨人乳头状瘤病毒(HPV)负荷量与子宫颈癌及其癌前病变的关系。方法:2003年3月~2003年12月在我院行宫颈癌筛查的2 651例患者为研究对象,采用杂交捕获2代(HCⅡ)方法定量检测宫颈HPV DNA的含量(以此表示宫颈HPV负荷量)。按HPV DNA含量分成5组,分别为:HPV<1,1≤HPV≤100,101≤HPV≤500,501≤HPV≤1 000,HPV>1 000。对其中HPV DNA阳性妇女,随后进行阴道镜检查、宫颈多点活检和颈管诊刮术。结果:794例阳性患者中的783例进行了阴道镜下多点活检和细胞学诊断,其中子宫颈炎650例(83.23%),C IN 102例(13.47%),宫颈癌26例(3.3%)。650例对照组、102例癌前病变组和26例宫颈癌组中,3组的HPV DNA定量总体分布有显著差别(P<0.005)。在HPV-DNA定量>100时,宫颈癌前病变的阳性率显著升高(P<0.005);当HPV-DNA的定量>500时,宫颈癌的阳性率显著升高(P<0.001)。C INⅡ、C INⅢ及宫颈癌在HPV DNA定量>1时的诊断阳性率较显著<1时显著高(P<0.01)。结论:HCⅡ方法是检测宫颈HPV感染的有效手段,HPV DNA含量随宫颈病变的严重程度而增加,可作为宫颈癌前病变及宫颈癌的筛查及病情追踪监测的1项客观指标。