体内电穿孔在质粒介导的基因转移及DNA免疫中的应用

Objective To investigate the effects of in vivo electroporation on plasmid mediated reporter gene expression and immunogenicity of DNA vaccine. Methods Luciferase expression plasmid was administered intramuscularly to BALB/c mice at 8μg and 40μg dosage level through injection with or without eletroporation Luciferase expression level in murine muscle was detected by IVIS imaging system 24 h after injection. DNA vaccine plasmid p1.0-gp1455m carrying codon-optimized env gene of CN54 strain ( HIV-1 CRF07_BC) was administered to mice at dosages of 8μg and 40μg through the two approaches mentioned above. Mice were immunized at week 0,2 and 4. Env-specific immune responses were detected at two weeks post the second and the third vaccinations. Env-specific antibody immune responses were determined by ELISA. Euv-specific cellular immune responses were determined by IFN-γ ELISPOT. Results Luciferase expression level in murine muscle was significantly increased as much as 35 folds through in vivo eletroporation. Results of ELISA and ELISPOT revealed that in vivo eletroporation could significantly enhance both the humoral and cellular immune responses induced by DNA vaccination. The responses induced by electrodelivered p1.0-gp1455m at 8 μg dosage were better than those induced by simple intramuscular injection with 40 μg of plasmid DNA. On the other hand, 2 injections followed by electroporation elicited comparable level of humoral and cellular immune responses with those induced by 3 injections without electroporation. Conclusion In vivo electroporation was capable of enhancing both the plasmid-mediated gene expression and immunogenicity of DNA vaccine.     

静脉吸毒者丙型肝炎病毒感染的研究

目的 进一步研究静脉吸毒者（ＩＶＤＵＳ）＿中丙型肝炎病毒（ＨＣＶ（）感染的特点。方法 采用ＥＬＩＳＡ法对１１４份ＩＶＤＵＳ,其中包括人类免疫缺陷病毒（ＨＩＶ）阳性５０份,ＨＩＶ阴性６４份,以３主１１８份非静脉吸毒者（ＮＩＶＤＵＳ）血清进行抗ＨＣＶ检测;并采用ＲＴ－ＰＣＲ法对其中抗ＨＣＶ阳性者和４０例抗ＨＣＶ阳性的非吸毒者血清用ＨＣＶ２型引物进行ＨＣＶ０ＲＮＡ检测。结果 ＨＩＶ阳性的ＩＶＤＵＳ抗ＨＣ     

福建省HIV-l流行毒株的基因序列测定和亚型分析

目的　通过DNA序列分析 ,确定福建省HIV 1流行毒株的亚型。方法　从福建省HIV 1抗体阳性者中选择 9例经性接触感染HIV 1者 ,采集其全血分离周围血淋巴细胞 (PBMCs)提取DNA作为PCR扩增的模板 ,设计合成 3对套式引物扩增HIV 1前病毒DNA的C2～V3区用于测序。所测序列与HIV 1中的 6个亚型的 10个国际标准或参考序列进行比较 ,确定被检标本的型别。结果使用PCR技术对 9份福建HIV 1阳性感染者PBMC样品进行扩增 ,获得HIV 1膜蛋白 (env)基因的核酸片段 ,并对其约 2 47bp核苷酸序列进行了分析、比较 ,证实 9份样品中 7份为E亚型 ,其余 2份分别为B和C亚型毒株。E亚型各株间序列存在一定差异 ,组内基因离散率为 6 2 8± 2 40。结论　福建省目前HIV 1的流行株主要为E亚型 ,流行株的来源较为复杂 ,株间序列差异不仅与感染时间有关 ,也和感染地点不同等因素有关 ,这些特点不同于我国其他一些省份发现的经血传播的HIV 1B或C亚型 ,其流行时间不长 ,株间序列差异不大     

以社区为基础的静脉吸毒人群共用注射器具行为特征研究

目的 了解凉山彝族自治州某地静脉吸毒人群共用注射器具的行为特征。方法 以社区为基础调查静脉吸毒人群人口学和共用注射器具吸毒行为情况。结果 在静脉吸毒的379人中，曾经共用注射器具的247人(65．2％)；近3个月使用别人用过的注射器具吸毒的166人(43．8％)。本人静脉注射转变到与他人共用注射器具吸毒的平均时间为363．73天，其中87人(35．2％)首次静脉注射吸毒即与他人共用注射器具。彝族吸毒人群从本人静脉吸毒转变到与他人共用注射器具吸毒的时间短，与他人共用注射器具吸毒的次数，以及近3个月使用别人用过的注射器具吸毒的次数，高于其他吸毒人群。低文化水平吸毒人群从本人静脉吸毒转变到与他人共用注射器具吸毒的时间短，与他人共用注射器具吸毒的次数，以及近3个月使用别人用过的注射器具吸毒的次数，高于其他吸毒人群。结论 针对彝族和低文化水平吸毒人群开展干预，以改变共用注射器具静脉吸毒的高危行为。     

艾滋病和艾滋病病毒感染诊断标准

1范围本标准规定了艾滋病和艾滋病病毒感染的诊断标准。本标准适用于全国各级医疗卫生机构和人员对艾滋病和艾滋病病毒感染的诊断和报告。2术语和定义下列术语和定义适用于本标准。2.1艾滋病病毒,人免疫缺陷病毒human immunodeficiency virus HIV是导致艾滋病的病原体。     

TZM-b1细胞系检测我国HIV-1病毒表型耐药性

目的 利用TZM-b1细胞系检测艾滋病病毒Ⅰ型(HIV-1)对逆转录酶抑制剂(AZT、d4T、ddI、NVP)的药物敏感性,并用该方法检测中国抗病毒治疗失败者的表型耐药.方法 用两种实验室适应毒株SF33和ⅢB来评价TZM-b1细胞方法的敏感性;用外周血单个核细胞(PBMC)共培养法分离扩增抗病毒治疗失败者的临床毒株;用TZM-b1法检测16例临床毒株对逆转录酶抑制剂的表型耐药.结果 AZT、d4T、ddI、NVP对SF33/ⅢB毒株的抑制50%病毒复制的药物浓度(IC50)分别为0.009/0.005μmol/L、0.125/0.113μmol/L、0.495/1.905μmol/L、0.093/0.045μmol/L.16例临床分离毒株中分别有13(81.3%)株、13(81.3%)株对AZT和NVP耐药.其中7株(53.8%)显示对AZT高度耐药;对NVP耐药的均为高度耐药,其中8株IC50升高1 000倍以上.结论 TZM-b1细胞系可用来快速检测HIV-1病毒表型耐药.我国抗病毒治疗失败者的表型耐药主要表现为对AZT和NVP耐药.     

中国南方四省区流行的HIV-1 CRF01_AE病毒株基因特征研究

Objective To analyze the genetic characteristics of HIV-1 CRF01_AE strains prevailing in. the four provinces of southern China. Methods Plasma samples were collected from the newly diagnosed HIV-1 individuals reported in 2006 in Guangdong, Guangxi, Jiangxi and Hunan province. The gag and env gene fragments were amplified from RNA template extracted from plasma using RT and nested PCR methods. CRF01_AE sequences were analyzed by phylogcnetie methods and characterized by calculating the genetic distance and Entropy analysis. Results Two main epidemic clusters were found to exist in the CRF01 AE strains from 210 HIV-1 CRF01 AE infected individuals collected in the 4 provinces, southern China. It was found that no international reference strain was closely correlated with cluster Ⅰ , which including 123 samples. The strains in cluster Ⅱ, consisting 57 cases of samples, were closely related with the strains identified in Vietnam. Genetic distance analysis of gag and env genes showed that the diversity of cluster Ⅰ was obviously less than that of cluster Ⅱ. Data on nucleotide polymorphism showed that nucleotides compositions of 42 sites in gag and 40 sites in env wer esignificantly different between the two clusters. When compared with cluster Ⅱ , the polymorphism decreased at 61 nucleotide sites but increased at 21 sites in cluster Ⅰ. Conclusion This was the first report describing that two main epidemic clusters were existed in CRF01_AE strains prevailing in the 4 provinces, Southern China. The vires in cluster Ⅰ was the dominant strain in this region, with shorter period of circulation and higher proportion seen in the HIV-infected population, which might belong to CRF01_AE strain with certain features facilitating the spread of the virus. The virus in cluster Ⅱ was highly homology with the CRF01_AE strains from Vietnam, and seemed to have had several events of epidemics in populations in border regions of China and Vietnam.     

四川省西昌市吸毒人群HCV感染及其危险因素研究

目的　了解四川省西昌市吸毒人群吸毒方式及性行为特征与HCV感染的关系。方法　于2 0 0 4 - 0 5 / 0 7,调查四川省西昌市吸毒人群的社会人口学、吸毒方式及性行为特征与HCV感染的关系。采集血样进行HCV和梅毒抗体检测。结果　在调查的吸毒人群6 19人中,HCV感染率为5 5 .9% (346 / 6 19) ,其中静脉吸毒者的HCV感染率为71 9% (2 35 / 32 7)。在单因素和多因素分析中,曾经静脉注射吸毒(OR =2 .6 3;95 %CI为1 .79～3 .89,P <0 .0 0 1)和曾经共用针头或注射器具吸毒(OR =2 .98,95 %CI为1 .78～5 . 0 0 ,P <0 . 0 0 1)与HCV感染有统计学意义。结论　吸毒人群HCV感染与其高危吸毒行为有关。     

共用注射器静脉吸毒与HIV感染关系的研究

目的探讨吸毒人群共用注射器静脉吸毒与HIV感染的关系。方法于2004-05/07,采用横断面研究调查四川省西昌市吸毒人群的社会人口学、吸毒频率、共用注射器静脉吸毒的次数和伙伴人数以及性行为特征。同时采集血样进行HIV和抗体检测。结果调查吸毒人群451人,HIV感染率为15.1%(68/451),其中静脉吸毒人群HIV感染率为17.8%(66/370)。在多因素Logistic回归模型分析中,与HIV感染关系有统计学意义的变量是彝族(OR=2.91;95%CI,1.67～5.10)、至今共用注射器具静脉吸毒的总次数(1～19次OR=4.67;95%CI,2.35～9.26;20次及以上OR=4.20;95%CI,1.52～11.61)和至今共用注射器具静脉吸毒的伙伴人数7人及其以上(OR=2.17;95%CI,1.01～4.68)。结论吸毒人群共用注射器具静脉吸毒的社会网络大小与其HIV感染有关,需通过降低共用注射器具静脉吸毒的次数和伙伴人数来控制HIV的传播流行。     

四川省西昌市吸毒人群首次海洛因滥用情况调查

目的 了解四川省西昌市吸毒人群首次海洛因滥用的发生情况。方法 于2004年5～7月，从社区中搜集吸毒人群，调查其社会人口学特征、首次使用毒品的时间和方式及种类、首次吸毒前吸烟、饮酒、朋友、家庭成员和亲戚吸毒的情况。结果 在调查的451名吸毒人员中，首次使用的毒品均为海洛因，其中，首次吸毒方式为口吸和静脉注射的分别为80．7％和19．3％。从出生到首次吸毒的发生率为4．43／100人年（95％CI=4．02～4．84）。多因素Cox比例风险模型分析结果显示，彝族及其他少数民族（FIR：1．47，95％CI=1．22～1．78）、15岁以前开始吸烟（FIR=1．54，95％CI=1．25～1．83）与首次吸毒发生的关系差异有统计学意义。结论 需进一步了解吸毒人群首次吸毒的发生情况及其影响因素，为开展针对性的健康教育和行为干预措施提供科学指导。