四川地区宫颈癌组织HPV18和HPV45 E6基因突变分析

采用聚合酶链反应技术对四川地区2003~2004年收集的60例宫颈癌患者的癌组织DNA进行人乳头瘤病毒(HPV) E6基因扩增, 获得HPV18和45型E6基因. 序列分析发现, 18型三例E6基因有同样的两处同义突变; 45型两例E6基因发生突变, 一例有两处碱基突变, 另一例发生六处碱基突变, 其中两处涉及氨基酸变化, 均位于E6抗原决定簇区. HPV45型E6基因中134位c→t, 157位c→t, 259位g→t和341位t→c的碱基点突变未见报道. 另外， 该地区HPV18和45型突变株之间存在碱基互变,  它们之间的最小差异比野生型HPV18和HPV45之间的差异小4.05%, 该数值比在非洲发现的突变株的要小很多, 该结果支持HPV18和HPV45可能起源于非洲的观点.     

Thermosensitive mucoadhesive gel formulation loaded with 5-Fu: cyclodextrin complex for HPV-induced cervical cancer

Human Papilloma Virus (HPV) infections are the major cause of cervical cancers. To achieve a better therapeutic efficacy and patient compliance in the treatment for HPV-induced cervical cancers, anticancer agent 5-fluorouracil has been formulated in a vaginal gel using the thermosensitive polymer Pluronic^® F127 together with alternative mucoadhesive polymers e.g., hyaluronic acid, Carbopol 934 and hydroxypropylmethylcellulose. To increase its aqueous solubility and to achieve the complete release of 5-FU from the gel, the drug was incorporated as its inclusion complex with 1:1 molar ratio with either β-cyclodextrin or hydroxypropyl-β-cyclodextrin. Following the characterization of drug:CD complexes, thermosensitive gel formulations containing different mucoadhesive polymers and the drug in free or complexed form were characterized in vitro by determining the gelation temperature and the rheological behavior of different formulations along with the in vitro release profiles of these formulations in pH 5.5 citrate buffer. It was observed that complexation with cyclodextrin accelerated the release of 5-FU with the exception of formulation containing Carbopol 934 as mucoadhesive polymer. As far as rheological properties are concerned, favorable thermosensitive in situ gelling properties were obtained with formulations containing HPMC as mucoadhesive polymer. Complete release of 5-FU from gels were obtained with both complexes of β-CD and HP-β-CD and cytotoxicity studies against HeLa human cervical carcinoma cells demonstrated that 1% 5-FU:CD complexes were equally effective as 1% free 5-FU indicating better therapeutic efficacy with lower dose.     

U.S. EPA Regulatory Perspectives on the Use of QSAR for New and Existing Chemical Evaluations

Abstract

As testing is not required, ecotoxicity or fate data are available for ≈ 5% of the approximately 2,300 new chemicals/year (26,000 + total) submitted to the US-EPA. The EPA's Office of Pollution Prevention and Toxics (OPPT) regulatory program was forced to develop and rely upon QSARs to estimate the ecotoxicity and fate of most of the new chemicals evaluated for hazard and risk assessment. QSAR methods routinely result in ecotoxicity estimations of acute and chronic toxicity to fish, aquatic invertebrates, and algae, and in fate estimations of physical/chemical properties, degradation, and bioconcentration. The EPA's Toxic Substances Control Act (TSCA) Inventory of existing chemicals currently lists over 72,000 chemicals. Most existing chemicals also appear to have little or no ecotoxicity or fate data available and the OPPT new chemical QSAR methods now provide predictions and cross-checks of test data for the regulation of existing chemicals. Examples include the Toxics Release Inventory (TRI), the Design for the Environment (DfE), and the OECD/SIDS/HPV Programs. QSAR screening of the TSCA Inventory has prioritized thousands of existing chemicals for possible regulatory testing of: 1) persistent bioaccumulative chemicals, and 2) the high ecotoxicity of specific discrete organic chemicals.     

Designing a potent L1 protein-based HPV peptide vaccine: A bioinformatics approach

BackgroundOncogenic human papilloma viruses (HPV) are the cause of various types of cancer, specifically cervical cancer. L1 protein is the main protein of HPV capsid which targeted in many vaccine-producing attempts. However, they have not enough coverage on the various high risk HPV types. Therefore, having a low cost potent HPV vaccine to protect against all members of the α-papillomaviridea family will be promising. In this study, L1 protein-based peptide vaccine was designed using immunoinformatics methods which provides physicochemical properties such as stability in room temperature, potential of antigenicity, non-allergic properties and no requirement with eukaryotic host system.ResultsThe designed vaccine has two HPV conserved epitopes with lengths 18 and 27 amino acids in all members of α-papillomaviridea. These peptides promote humoral and cellular immunity and INF-γ responses. In order to ensure strong induction of immune responses, Flagellin, a Toll like receptor 5(TLR-5) agonist, and a short synthetic toll like receptor 4 (TLR-4) agonist were also joined to the epitopes. Structure of the designed- vaccine was validated using Rampage and ERRAT and a high quality 3D structure of the vaccine protein was provided. Docking studies demonstrated an appropriate and stable interaction between the vaccine and TLR-5.ConclusionsThe vaccine is expected to have a high quality structure and suitable properties including high stability, solubility and a high potential to be expressed in E.coli. High potentiality of the vaccine in inducing humoral and cellular immune responses, may be considered as an anti-tumor vaccine.     

Impact of Collection Volume and DNA Extraction Method on the Detection of Biomarkers and HPV DNA in First-Void Urine

The potential of first-void (FV) urine as a non-invasive liquid biopsy for detection of human papillomavirus (HPV) DNA and other biomarkers has been increasingly recognized over the past decade. In this study, we investigated whether the volume of this initial urine stream has an impact on the analytical performance of biomarkers. In parallel, we evaluated different DNA extraction protocols and introduced an internal control in the urine preservative. Twenty-five women, diagnosed with high-risk HPV, provided three home-collected FV urine samples using three FV urine collection devices (Colli-Pee) with collector tubes that differ in volume (4, 10, 20 mL). Each collector tube was prefilled with Urine Conservation Medium spiked with phocine herpesvirus 1 (PhHV-1) DNA as internal control. Five different DNA extraction protocols were compared, followed by PCR for GAPDH and PhHV-1 (qPCR), HPV DNA, and HBB (HPV-Risk Assay), and ACTB (methylation-specific qPCR). Results showed limited effects of collection volume on human and HPV DNA endpoints. In contrast, significant variations in yield for human endpoints were observed for different DNA extraction methods (p < 0.05). Additionally, the potential of PhHV-1 as internal control to monitor FV urine collection, storage, and processing was demonstrated.     

甘肃地区女性宫颈HPV感染的现状研究

目的:了解甘肃地区女性人乳头瘤病毒(HPV)感染的基因亚型、年龄及地区分布等.方法:采用凯普医用核酸分子快速导流杂交基因芯片技术,对7318名兰州女性进行生殖道21种HPV感染基因亚型筛查.结果:甘肃地区女性HPV感染率为19.9%(1 452/7 318),其中高危型HPV感染率14.78%(1 082/7 318),低危型HPV感染率3.38%(247/7 318),高危型中HPV16为主要致病亚型,占28.59%(415/1 452),低危型中HPV6占6.30%(98/1 452).1452例感染HPV的标本中,单一感染1 188例,占81.82%(1 188/1 452),多重感染264例,占18.18%(264/1 452).按年龄分组中≥50岁组HPV感染率最高,占24.24%.按地区分析,陇南地区HPV感染率最高,达到30.97%(83/268).结论:甘肃地区为宫颈癌高发区,其中HPV16是感染最多的型别,对该地区女性进行子宫颈癌的筛查和防治已成为当务之急.     

宫颈HPV感染与宫颈瘤样病变的相关性

为了研究湘西地区妇女宫颈人乳头瘤病毒（HPV）感染与宫颈瘤样病变的相关性,为宫颈癌的早期预防提供科学参考,选取692例湘西籍患者为研究对象,所有患者均行宫颈HPV检测,以其中HPV检查结果阳性的234例患者作为观察组,阴性的234例患者作为对照组,均在阴道镜监视下进行宫颈多点活检及宫颈管搔刮病理学检查,病理检测结果分为癌前病变（CINⅠ,Ⅱ,Ⅲ）、宫颈癌、慢性宫颈炎.结果显示：湘西地区女性中HPV感染以HPV16（25.64%）类型最多,其余依次为HPV52,HPV51,HPV58,HPV18,HPV53为常见;CIN的加重程度随HPV感染率升高,CINⅠ,Ⅱ,Ⅲ两两比较差异具有统计意义;HPV阳性率为33.81%,HR-HPV（+）31.74%,其中单一感染HR-HPV（+）21.97%,双重或多重感染11.99%,LR-HPV（+）感染1.88%.说明在宫颈癌的筛查中高危HPV检查具有临床价值,多途径切断高危HPV感染对本地区妇女宫颈癌的一级预防具有重要意义.     

Development of interdigitated arrays coated with functional polyaniline/MWCNT for electrochemical biodetection: application for human papilloma virus

In this study, polyaniline-multiwalled carbon nanotube film (PANi-MWCNT) has been polymerized on interdigitated platinum electrode arrays (IDA), fabricated by MEMS technology for the detection of human papillomavirus (HPV) infection, using immobilized peptide aptamers as affinity capture reagent. Label-free, electrochemical detection of the specific immune reaction between antigen peptide aptamer HPV-16-L1 (with a molecular weight of 1825 Da), the most common genotype in cytological normal women worldwide, and its specific antibody of HPV-16 (which is much bigger with molecular weight of ca. 150 kDa) on multifunctional PANi-MWCNT based arrays was reported. The most significant advantage of this technique consists of reagentless and multiple detection of antigen-antibody complex formation on well conducting IDA interface of PANi-MWCNT, without intermediate steps or any labeling reagents, as normally required in the previous works.     

Preliminary report on detection of papillomaviruses types 6, 11, 16, and 18 in laryngeal benign and malignant lesions

Recently, clinicians have observed an increase in the incidence of laryngeal papilloma coincident with the rising incidence of venereal warts. The lesions occur in young adults of professional singing age. Different types of human papillomavirus (HPV) are presented. Gynecologic exam revealed cervical tumors, involving 18 HPV in benign and malignant lesions. Thirty patients with the following pathologic conditions were studied: 10 papillomas, 10 severe dysplasias, and 10 invasive cancers. All the lesions were located on the free edge of the vocal folds or the anterior commissure involving the epithelium. The results are as follows: 11 patients (33%) had virus infection, of which four had HPV 6 or HPV 11 on laryngeal papilloma, three had HPV 16 or HPV 18 on severe dysplasia, four had HPV 16 or HPV 18 on carcinoma; and 19 (66%) had no virus infection. Gynecologic research in this field is more advanced. However, this preliminary report raises interesting research questions in laryngology.     

Evaluating the potential of vaccine-induced type replacement for high-risk human papillomaviruses

Persistent infection with human papillomavirus (HPV) is the main cause of cervical cancer. Current HPV vaccines protect against both HPV-16 and -18, which are known to cause approximately 70% of cervical cancer cases worldwide. These vaccines have shown to be highly effective in preventing infection by their targeted types. However, there is a broad diversity of HPV types not targeted by the vaccines, and there is controversy about a possible increase in the prevalence of these non-targeted types after a vaccination program. Here, we propose a within-host metapopulation model to study the possibility of vaccine-induced type replacement for oncogenic types. It is generally believed that the theoretical possibility of type replacement strongly depends on the existence of natural type competition mechanisms. Nevertheless, our results suggest that type replacement is viable at the within-host level if the degree of cross-protection induced by the vaccine is low, even if there is no underlying competition among HPV types. Consequently, the impact of current HPV vaccines at both the immunological and epidemiological levels rely upon the level of cross-protection.