新疆维族妇女感染的HPV 型别分布的研究

目的:探讨人乳头瘤状病毒(Human papillomavirus,HPV)在新疆南部维族妇女人群中的型别分布情况。方法:以年龄在30-59岁的新疆伽师县夏普吐勒乡维吾尔族妇女人群为基础进行筛查,签署知情同意书后,采集受试者宫颈脱落细胞,利用PCR和基因芯片技术检测HPV DNA并分型。结果:共2473名妇女入选。HPV总的感染率为9.1%,高危型中HPV-16的感染率最高为6.9%,其他高危型的感染率从高到低依次为:HPV-59、HPV-56、HPV-18、HPV-33、HPV-58、HPV-51、HPV-31、HPV-45、HPV-52、HPV-68、HPV-35、HPV-39。低危型中HPV11感染率最高,其他低危型的感染率从高到低依次为HPV-42、HPV-43、HPV-6、HPV-53、HPV-66、HPV-73。HPV-44、-83、-MM4没检测到。多重感染率为34.2%。结论:新疆维吾尔族妇女人群中以HPV16感染为主,其次为HPV59、56、18、33等。HPV59可能是新疆维吾尔族妇女较易感染的类型。体现了新疆维吾尔族妇女感染HPV的特殊性。     

宫颈癌组织人乳头瘤病毒的荧光偏振基因分型

采用荧光偏振人乳头瘤病毒(human papillomavirus,HPV)分型新方法探讨了8种常见型别HPV在陕西宫颈癌患者中的流行情况。首先,用HPV GP5 ／GP6 通用引物PCR扩增65例早期宫颈癌(Ⅱa期内)和72例慢性宫颈炎病变组织DNA粗提物,继之将模板指导的末端延伸反应与荧光偏振检测技术结合(TDI-FP),用GP5 ／GP6 扩增区内的HPV6、11、16、18、31、33、35和58型特异性探针与PCR产物杂交后,荧光素标记的特异碱基(TAMRA-ddTTP或R110-ddGTP)在GP5 ／GP6 产物中相应的模板指导下,掺入延伸至相应探针末端,致使对应的TAMRA或R110 FP值升高,从而对扩增的HPV阳性产物进行HPV分型。65例宫颈癌患者中检出HPV57例,阳性率87．69％,72例慢性宫颈炎患者中检出HPV28例,阳性率38．89％,两组间HPV阳性率有显著性差异。宫颈癌与慢性宫颈炎患者中4种最常见的HPV型别分别是HPV 16(45．6％)、HPV 18(22．8％)、HPV 58(17．5％)、HPV 31(7．02％)和HPV 16(35．7％)、HPV 11(32．1％)、HPV 6(21．4％)、HPV 18(10．7％)。慢性宫颈炎患者中检出的HPV型别57．14％属高危型。HPV 16在两组中均最为多见。中国陕西宫颈疾病患者中HPV感染有其特点,世界范围内少见的HPV 58在陕西宫颈癌与慢性宫颈炎患者中均较为多见,在进行HPV新诊断方法及疫苗研制时应考虑到这种特点。     

人乳头瘤病毒HPV31型宫颈癌细胞模型的建立

宫颈癌与人乳头瘤病毒感染密切相关,建立宫颈癌实验模型可为宫颈癌的研究提供理想的模拟实验条件。我们通过应用基因重组技术,分别以HPV31型E6和E7基因为目的基因,通过原核表达、蛋白纯化和免疫小鼠等获得其特异性检测抗体。我们还通过构建E6和E7基因真核表达载体、转染C33A细胞、博莱霉素抗性筛选和表达检测等步骤,获得一种稳定的体外宫颈癌细胞系。经酶切鉴定及测序证实细胞基因组已重组插入质粒中的目的基因。我们已成功筛选到稳定的目的mRNA和蛋白表达的阳性细胞系,建立了稳定的人乳头状瘤病毒31型(HPV31)的宫颈癌细胞株,为研究宫颈癌提供了体外实验模型。     

Some Novel Adenosine Mimics: Synthesis and Anticancer Potential against Cervical Cancer caused by Human Papilloma Virus

Two novel adenosine analogs, viz. 9-(1′-β-D-arabinofuranosyl)-6-nitro-1,3-dideazapurine or Ara-NDDP (1) and 9-(5′-deoxy-5′-S-(propionic acid) (1′-β-D-ribofuranosyl) adenine or SAH analog (2), indigenously synthesized, have been found to be potential anticancer agents against cervical cancer caused by human papilloma virus.     

Age at start of using tobacco on the risk of head and neck cancer: Pooled analysis in the International Head and Neck Cancer Epidemiology Consortium (INHANCE)

BackgroundTobacco use is a well-established risk factor for head and neck cancer (HNC). However, less is known about the potential impact of exposure to tobacco at an early age on HNC risk.MethodsWe analyzed individual-level data on ever tobacco smokers from 27 case-control studies (17,146 HNC cases and 17,449 controls) in the International Head and Neck Cancer Epidemiology (INHANCE) consortium. Adjusted odds ratios (ORs) and 95% confidence intervals (CIs) were estimated using random-effects logistic regression models.ResultsWithout adjusting for tobacco packyears, we observed that younger age at starting tobacco use was associated with an increased HNC risk for ever smokers (OR_{<10 years vs. ≥30 years}: 1.64, 95% CI: 1.35, 1.97). However, the observed association between age at starting tobacco use and HNC risk became null after adjusting for tobacco packyears (OR_{<10 years vs. ≥30 years}: 0.97, 95% CI: 0.80, 1.19). In the stratified analyses on HNC subsites by tobacco packyears or years since quitting, no difference in the association between age at start and HNC risk was observed.ConclusionsResults from this pooled analysis suggest that increased HNC risks observed with earlier age at starting tobacco smoking are largely due to longer duration and higher cumulative tobacco exposures.     

5434例女性泌尿生殖道HPV感染情况及HPV亚型分析

目的了解泰顺地区女性易感人群人乳头瘤病毒(HPV)感染率和HPV亚型分型情况。方法收集2017年1月至2017年12月于泰顺县人民医院体检及就诊的女性患者泌尿生殖道标本,共5434例。采用PCR反向点杂交法进行HPV分型检测,并对结果进行分析。结果5434例标本中HPV阳性1895例,HPV阳性1309人次。高危型HPV检出1127例,占59.4%;低危型HPV检出356例,占18.8%。入选患者中单一感染935例,占71.4%;多重感染374例,占28.6%。高危型中检出最多的是HPV52型,共210例,占11.1%;其次是HPV58型137例,占7.2%;HPV16型124例,占6.5%;HPV18型62例,占3.3%。HPV52阳性患者集中于41~50岁,HPV53阳性患者集中于31~40岁,HPV16、HPV18阳性患者集中于20~40岁及>60岁。随着患者年龄的增长各HPV亚型的阳性率也在增长,且各年龄段患者均以高危型HPV感染为主(62.36%)。结论泰顺地区女性患者HPV亚型及患者年龄分布情况与我国南方女性HPV感染情况相符,HPV感染率随患者年龄增加而增高。高危型HPV检测对子宫颈癌等相关疾病的筛查及预防具有较高的应用价值。     

人乳头瘤病毒16E6基因通过增加HMGB1表达调节口腔癌CAL27细胞侵袭的实验研究

人乳头瘤病毒16型(Human papillomavirus type 16,HPV16)感染与口腔癌、宫颈癌的发病有关,HPV16 E6基因编码的蛋白是重要的癌蛋白,已经被证实能够通过增加高迁移率族蛋白B1(High mobility group box-B1,HMGB1)表达来促进宫颈癌细胞的侵袭,但是否能调控口腔癌细胞的侵袭仍未明确。为研究HPV16 E6基因通过增加HMGB1表达调节口腔癌CAL27细胞侵袭的作用,口腔癌CAL27细胞被分为对照组、空白质粒组、HPV16 E6质粒组、NC-si RNA组(短片断干扰RNA阴性对照组)、NC-si RNA+HPV16 E6质粒组、HMGB1-si RNA+HPV16E6质粒组,检测细胞中HPV16 E6及HMGB1的表达、细胞的侵袭数目、培养基中HMGB1的含量。结果显示,HPV16 E6质粒组细胞中HPV16 E6及HMGB1的表达量、培养基中HMBG1的含量、细胞的侵袭数目均高于对照组及空白质粒组(P<0.05);HMGB1-si RNA组细胞中HMGB1的表达量明显低于对照组及NC-si RNA组(P<0.05);NC-si RNA+HPV16 E6质粒组的细胞侵袭数目均明显高于NC-si RNA组(P<0.05),HMGB1-si RNA+HPV16 E6质粒组的细胞侵袭数目均明显低于NC-si RNA+HPV16 E6质粒组(P<0.05)。本研究提示,HPV16 E6基因能够促进口腔癌CAL27细胞的侵袭且这一作用与增加HMGB1表达有关。     

Pin1，CyclinD1在宫颈癌中的表达及与HPV16／18感染的关系

研究肽基脯氨酰顺反异构酶Pin1,CyclinD1在宫颈液基细胞上皮内病变和宫颈癌中的表达与HPV16/18感染的关系,探讨其对宫颈上皮内病变和宫颈癌诊断的意义。经液基细胞学筛查,采用原位杂交方法对80例宫颈上皮内病变和宫颈癌,13例正常组织进行HPV检测分型,同时对HPV16/18阳性的标本进行Pin1,CyclinD1免疫组化。在正常宫颈组织、CINⅠ,CINⅡ-Ⅲ,浸润癌组织中Pin1表达率分别为7.7%(1/13)、57.1%(12/21)、68.4%(13/19)、85%(34/40)(P<0.05);CyclinD1表达率分别为0%、9.52%(2/21)、20.5% (4/19)、55%(22/40)。HPV16/18的表达率也随着宫颈病变的升级而升高,分别为15.4%(2/13)、42.9%(9/ 21)、57.9%(11/19)、82.5%(33/40)。Pin1和HPV16/18在宫颈病变中的表达高度相关(r_0=1.0,P<0.05);而CyclinD1的表达与HPV16/18相关性较小(r_0=0.4,P<0.05)。认识Pin1,cyclinD1和HPV16/18三者之间的关系及对宫颈上皮内病变和宫颈癌的筛查诊断具有重要意义。     

宫颈癌患者人乳头瘤病毒(HPV)主要型别及其感染研究

本文探讨了江西省和广东省宫颈癌患者人乳头瘤病毒(Human papillomavirus,HPV)感染及其型别分布,分析了高危型HPV对各种宫颈病变的感染情况,为宫颈癌的早期发现和临床诊治提供科学依据。首先采用细胞学、HPV DNA检测(第二代杂交捕获法,HC2)、电子阴道镜和宫颈化学着色方法筛查宫颈癌患者,经病理镜检确诊,然后用GP PCR-SBT法对宫颈癌患者进行HPV基因分型。江西省溪口镇、古市镇及修水县城宫颈癌癌前病变发生率为5．7‰。HC2方法发现宫颈癌患者13种高危型HPV DNA阳性率为89．9％,宫颈上皮内瘤样病变的为84．8％,对照组为24．5％。采用GP PCR-SBT方法进行基因分型发现,江西省宫颈癌患者存在HPV16、58、31、33、18、66、6、11、56和81十种型别,其中HPV81型在国内外鲜有报道。据此提出生殖道高危型HPV感染是妇女宫颈癌发病的重要因素。并发现江西省宫颈癌高发区妇女高危型HPV感染率为24．5％。建立了HPV基因分型的方法,对HPV致宫颈病变的分子机制进行了分析。     

反向点杂交法快速检测HPV基因型的临床应用

应用反向点杂交法(RDB)的原理,针对HPV 6B, 11, 16, 18, 31, 33和35设计了7条序列作为未标记的特异性寡核苷酸(SSO)探针,分别固定在尼龙膜条上,形成7个点,再与经PCR扩增的样品DNA序列杂交,即可在一个膜条上分辨出这7型HPV中的任一型.此法快速简便,特异性高,不存在假阳性;且因PCR灵敏度高,亦不易出现假阴性.用PCR-RDB法检测保存的宫颈癌组织石蜡包埋标本32例,结果:HPV16阳性22例(68.8%),HPV18阳性5例(15.6%),HPV16/18双重感染2例(6.3%),阴性仅3例(9.3%).     

Simultaneous Integrated Boost Radiotherapy in Unresectable Stage IV (M0) Head and Neck Squamous Cell Cancer Patients: Daily Clinical Practice

AimTo evaluate clinical outcome in locally-advanced stage IV (M0) head and neck cancer patients treated using intensity-modulated radiotherapy (IMRT) with simultaneous integrated boost (SIB) in daily clinical practice.BackgroundDespite SIB-IMRT has been reported as a feasible and effective advanced head and neck cancer treatment, there are few data about its concurrent use with systemic therapies.Material and MethodsWe reviewed 41 staged IV (M0) head and neck cancer patients treated in two radiotherapy units in the city of Messina (Italy) during the last six years, using intensity modulated techniques-SIB. 22/41 patients had concomitant chemotherapy or cetuximab. Acute and late toxicities, objective response (OR) rate, local control (LC) and overall survival (OS) have been evaluated.Results37/41 patients received the planned doses of radiotherapy, 2 patients died during the therapy. The major acute regional toxicities were skin reaction and mucositis. A case of mandibular osteoradionecrosis was recorded. At completion of treatment, OR was evaluated in 38 patients: 32/38 patients (84.2%) had complete (55.3%) and partial (28.9%) response. The 1- and 5-year LC rates were 73.4% and 69.73%, respectively. The 1-, 3-, and 5-year OS rates were 85.93%, 51.49% and 44.14%, respectively. No statistically significant differences in outcomes have been observed in patients treated with radiotherapy alone vs. irradiation concomitant to chemo/biotherapy. The median OS was 45 months.ConclusionSIB-IMRT is safeand can be used with concomitant chemotherapy/biotherapy in real-life daily clinical practice. SIB-IMRT alone is a valid alternative in patients unfit for systemic therapies.     

Immunization with a poly (lactide co-glycolide) encapsulated plasmid DNA expressing antigenic regions of HPV 16 and 18 results in an increase in the precursor frequency of T cells that respond to epitopes from HPV 16, 18, 6 and 11

A phase II trial was conducted in subjects with human papillomavirus (HPV) associated high-grade cervical dysplasia testing the safety and efficacy of a microparticle encapsulated pDNA vaccine. Amolimogene expresses T cell epitopes from E6 and E7 proteins of HPV types 16 and 18. An analysis was performed on a subset of HLA-A2+ subjects to test whether CD8+ T cells specific to HPV 16, 18, 6 and 11 were increased in response to amolimogene immunization. Of the 21 subjects receiving amolimogene, 11 had elevated CD8+ T cell responses to HPV 16 and/or 18 peptides and seven of these also had increases to corresponding HPV 6 and/or 11 peptides. In addition, T cells primed and expanded in vitro with an HPV 18 peptide demonstrated cross-reactivity to the corresponding HPV 11 peptide. These data demonstrate that treatment with amolimogene elicits T cell responses to HPV 16, 18, 6 and 11.     

Cervico-vaginal dysplasia--papillomavirus-induced and HIV-1 infection: role of correlated markers for prognostic evaluation

Sexually-transmitted diseases (STD) can facilitate the progression of HIV-1 infection. Among them, as we have previously demonstrated, cervico-vaginal dysplasia-papillomavirus (HPV)-induced, together with HSV-2 co-infection, seems to be correlated with a more evident immunodepression in HIV-positive women, compared with other sexually transmitted diseases. Here we have analysed some of the main correlated markers of HIV-1 infection progression: CD4 + T lymphocyte concentration, CD4 +/CD8 + T cells ratio, HIV-1 RNA loads and haemoglobin (Hb) concentration in 30 HIV-1 positive women co-infected with HPV, and suffering from cervico-vaginal dysplasia, in different stages. In particular, we noticed a positive correlation, evaluated by Spearman's test, between the degree of progression of dysplastic stages (CIN1 --> 3) until invasive carcinoma (IC) and HIV-1 RNA loads (C(s) = +0.78; p < 0.001), and in contrast, a negative correlation between the same stages of progression and respectively CD4 + T cell concentration (C(s) = -0.54; p = 0.01), ratio (C(s) = - 0.63; p = 0.002) and Hb concentration (C(s) = -0.85; p < 0.001). In conclusion, it is important to underline that low levels of Hb generally paralleled the degree of immunodepression. In fact CD4 + T cell levels and ratio positively correlated with Hb concentrations respectively, with C(s) = + 0.83; p < 0.001 and C(s) = + 0.90; p < 0.001. Finally, the most efficacious antiretroviral combined therapy (HAART = Highly Active Antiretroviral Therapy) can improve the above described laboratory parameters in HIV-1/HPV co-infected women and seems to prevent the progression of CIN1 to the following stages of the dysplastic disease.     

Human internal mammary artery (IMA) transplantation and stenting: a human model to study the development of in-stent restenosis

Preclinical in vivo research models to investigate pathobiological and pathophysiological processes in the development of intimal hyperplasia after vessel stenting are crucial for translational approaches (1,2). The commonly used animal models include mice, rats, rabbits, and pigs (3-5). However, the translation of these models into clinical settings remains difficult, since those biological processes are already studied in animal vessels but never performed before in human research models (6,7). In this video we demonstrate a new humanized model to overcome this translational gap. The shown procedure is reproducible, easy, and fast to perform and is suitable to study the development of intimal hyperplasia and the applicability of diverse stents. This video shows how to perform the stent technique in human vessels followed by transplantation into immunodeficient rats, and identifies the origin of proliferating cells as human.     

Biomarkers for early detection of high risk cancers: From gliomas to nasopharyngeal carcinoma

Nasopharyngeal carcinoma (NpC) is a malignant disease associated with Epstein-Barr virus infection, and often diagnosed at an advanced stage. This significantly curtails patient survival. We hypothesize that a panel of biomarkers can be assembled to assess NpC incidence, early detection, and tumor progression during therapeutic intervention. Our thesis rests on a model of successfully predicting high-risk gliomas by means of a carefully crafted panel of molecular mitotic biomarkers (i.e., securin, survivin and MCM2). The strategy we propose holds strong promise for prevention and cure of NpC. The approach we propose seeks to identify certain biomarkers from viral materials, patient tissues and assessment of related diseases, whose signatures, taken together, will be endowed with some degree of congruency, or sense of a coordinated language (i.e., “votes”). Biomarker “voting” will then permit to outline a broad coordinated molecular map for the molecular and epigenetic characterization of each individual patient''s NpC tumor. We will draw on the process of contrasting biomarkers in health and disease, which rests on the auto-proteomic concept particularly relevant in high-risk cancer individuals, such as is the case for NpC. In brief we defend, current advances in human proteome profiling proffers the possibility of having individual baseline proteomic profiles using local body fluids (e.g., saliva, nasal secretions, sputum) or systemic fluids (e.g., plasma, serum, cerebrospinal fluid) to unravel a personalized molecular map for high-risk NpC individuals. Regular check-up will monitor for new or impending manifestations of NpC, and provide a secure assessment of incidence and early detection.     

Clinical characteristics and molecular epidemiology of human metapneumovirus in children with acute lower respiratory tract infections in China, 2017 to 2019: A multicentre prospective observational study

Human metapneumovirus (HMPV) infection is one of the leading causes of hospitalization in young children with acute respiratory illness. In this study, we prospectively collected respiratory tract samples from children who were hospitalized with acute lower respiratory tract infection in six hospitals in China from 2017 to 2019. HMPV was detected in 145 out of 2733 samples (5.3%) from the hospitalized children. The majority of HMPV-positive children were under the age of two (67.6%), with a median age of one year. HMPV can independently cause acute lower respiratory tract infection in young children, while all patients showed mild clinical symptoms. Of all the co-infected patients, HMPV was most commonly detected with enterovirus (EV) or rhinovirus (RhV) (38.0%, followed by respiratory syncytial virus (RSV) (32.0%). The highest detection rate occurred from March to May in both northern and southern China. Out of 145 HMPV positive samples, 48 were successfully typed, of which 36 strains were subgrouped into subtypes A2c (75%), eight strains were included in subtype B1 (16.7%), and four strains were included in subtype B2 (8.3%). Moreover, 16 A2c strains contained 111-nucleotide duplications in the G gene. Twenty-seven complete HMPV genomes were successfully obtained, and 25 (92.6%) strains belonged to subtype A2c, whereas one strain was included in subgroup B1 and another was included in subgroup B2. A total of 277 mutations were observed in the complete genomes of 25 A2c strains. All results presented here improve our understanding of clinical characteristics and molecular epidemiology of HMPV infection in children.     

Binding of the Bovine and Porcine Pancreatic Secretory Trypsin Inhibitor (Kazal) To Human Leukocyte Elastase: A Thermodynamic Study

Abstract

The effect of pH and temperature on the apparent association equilibrium constant (K_a) for the binding of the bovine and porcine pancreatic secretory trypsin inhibitor (Kazal-type inhibitor, PSTI) to human leukocyte elastase has been investigated. At pH8.0, values of the apparent thermodynamic parameters for human leukocyte elastase: Kazal-type inhibitor complex formation are: bovine PSTT – K_a = 6.3 × 10⁴M^?1, δ5G° = -26.9kJ/mol, δH° = +11.7kJ/mol, and δS° = +1.3 × 10² entropy units; porcine PSTI –K_a = 7.0 × 10³M^?1,δG° = -21.5kJ/mol, δH° = +13.0kJ/mol, and δS° = +1.2 × 10² entropy units (values of K_a δG° and δS° were obtained at 21.0°C; values of δH° were temperature independent over the range (between 5.0°C and 45.0°C) explored). On increasing the pH from 4.5 to 9.5, values of K_a for bovine and porcine PSTI binding to human leukocyte elastase increase thus reflecting the acidic pK-shift of the His57 catalytic residue from ?7.0, in the free enzyme, to ?5.1, in the serine proteinase: inhibitor complexes. Thermodynamics of bovine and porcine PSTI binding to human leukocyte elastase has been analyzed in parallel with that of related serine (pro)enzyme/Kazal-type inhibitor systems. Considering the known molecular models, the observed binding behaviour of bovine and porcine PSTI to human leukocyte elastase was related to the inferred stereochemistry of the serine proteinase/inhibitor contact region(s).     

Nuclear patterns of human breast cancer cells during apoptosis: characterisation by fractal dimension and co-occurrence matrix statistics

An analytical strategy combining fractal geometry and grey-level co-occurrence matrix (GLCM) statistics was devised to investigate ultrastructural changes in oestrogen-insensitive SK-BR3 human breast cancer cells undergoing apoptosis in vitro. Apoptosis was induced by 1 μM calcimycin (A23187 Ca²⁺ ionophore) and assessed by measuring conventional cellular parameters during the culture period. SK-BR3 cells entered the early stage of apoptosis within 24 h of treatment with calcimycin, which induced detectable changes in nuclear components, as documented by increased values of most GLCM parameters and by the general reduction of the fractal dimensions. In these affected cells, morphonuclear traits were accompanied by the reduction of distinct gangliosides and loss of unidentifiable glycolipid molecules at the cell surface. All these changes were shown to be involved in apoptosis before the detection of conventional markers, which were only measurable during the active phases of apoptotic cell death. In overtly apoptotic cells treated with 1 μM calcimycin for 72 h, most nuclear components underwent dramatic ultrastructural changes, including marginalisation and condensation of chromatin, as reflected in a significant reduction of their fractal dimensions. Hence, both fractal and GLCM analyses confirm that the morphological reorganisation of nuclei, attributable to a loss of structural complexity, occurs early in apoptosis. This work was supported by grant 31-57626.99 from the Swiss National Science Foundation and grant FOR 450.94 from the Swiss Cancer League.     

Medicinal chemistry of indole derivatives: Current to future therapeutic prospectives

Indole is a versatile pharmacophore, a privileged scaffold and an outstanding heterocyclic compound with wide ranges of pharmacological activities due to different mechanisms of action. It is an superlative moiety in drug discovery with the sole property of resembling different structures of the protein. Plenty of research has been taking place in recent years to synthesize and explore the various therapeutic prospectives of this moiety. This review summarizes some of the recent effective chemical synthesis (2014–2018) for indole ring. This review also emphasized on the structure–activity relationship (SAR) to reveal the active pharmacophores of various indole analogues accountable for anticancer, anticonvulsant, antimicrobial, antitubercular, antimalarial, antiviral, antidiabetic and other miscellaneous activities which have been investigated in the last five years. The precise features with motives and framework of each research topic is introduced for helping the medicinal chemists to understand the perspective of the context in a better way. This review will definitely offer the platform for researchers to strategically design diverse novel indole derivatives having different promising pharmacological activities with reduced toxicity and side effects.