Recombinant glycoprotein based vaccine for Chandipura virus infection

Chandipura virus (CHPV) has emerged as an important pediatric encephalitis-causing pathogen with very high mortality in India. No specific vaccine or treatment is available till date. We attempted to prepare a candidate vaccine employing recombinant CHPV Glycoprotein (rGp). The Glycoprotein gene (G-gene) of CHPV was expressed using Baculovirus expression system. The rGp was purified by HPLC and used for mice immunization, 3 doses, and 4 weeks apart. One microgram rGp was found to be optimum. Sero-conversion was observed as early as 2nd week by detecting anti-CHPV IgG antibodies. Antibody titres were immunogen-concentration dependent. Intracerebral challenge of the immunized mice with 100 LD₅₀ of the homologous strain demonstrated 90% protection. In in vitro neutralization, antibodies from the immunized mice were able to neutralize heterologous viruses. There was 60% T cell proliferation observed against rGp in immunized mice. The study shows that rGp induces both arms of immune response and represents an ideal vaccine candidate for further evaluations.     

瘢痕疙瘩形成机制的研究进展

瘢痕疙瘩是烧伤外科研究的重要课题，其形成机制尚未完全明确，本文就其在遗传机制、免疫作用、细胞增殖与胶原代谢、细胞因子等方面的研究进展进行综述。     

血管内皮型一氧化氮合酶基因Glu298 Asp多态性与老年脑梗死的相关性研究

目的　通过病例对照研究 ,了解中国老年人群血管内皮型一氧化氮合酶 (eNOS)基因目的　通过病例对照研究 ,了解中国老年人群血管内皮型一氧化氮合酶 (eNOS)基因Glu2 98Asp多态性的分布 ,分别探讨其与老年脑梗死及血脂以及一氧化氮等脑梗死危险因素的关系。 方法对门诊及住院中确诊的 4 0例老年脑梗死和 16 9例性别、年龄相匹配的老人 ,测量他们的身高、体重及座位血压 ,并测定他们的空腹血脂、空腹血糖 (FBS)及一氧化氮 (NO)等 ,应用聚合酶链反应 (PCR)和限制性片断长度多态性 (RFLP)检测eNOS基因Glu2 98Asp多态性。结果　脑梗死和对照组eNOS基因Glu2 98Asp多态性构成有显著性差异 (χ2 =4 31,P =0 0 38) ,脑梗死组Glu/Asp基因型高于对照组(32 5 %vs 17 8% ) ;脑梗死组 2 98Asp等位基因频率高于对照组 (16 2 5 %vs 8 9% ) ,但是两组等位基因频率的分布比较 ,没有显著性差异 (χ2 =3 81,P =0 0 5 1)。结论　eNOS基因Glu2 98Asp多态性在中国老年人群中存在 ,并且基因...     

湖北人群Toll样受体4基因多态性研究

目的 研究湖北人群Toll样受体4(TLR4)Asp299Gly和Thr399Ile基因多态性分布特征。方法 采用等位基因特异性聚合酶链反应(ASPCR)-限制酶片断长度多态性(RFLP)检测TLR4 Asp299Gly和Thr399 Ile基因多态性。结果 在所有的253例中国湖北人群样本中，没有检测到TLR4 Asp299Gly和Thr399Ile基因多态性存在。结论 TLR4基因多态性在不同地区和人群发生的频率是不同的，与白种人相比，中国湖北人群的TLR4Asp299G1y和Thr399Ile的基因多态性非常罕见。     

间日疟原虫深圳株红内期SSUrDNA基因片段的克隆与序列分析

目的 体外扩增间日疟原虫深圳株红内期小亚单位核糖体核糖核酸编码基因(SSUrDNA)片段，研究其结构与功能。方法 设计一对特异性引物，采用聚合酶链反应(PCR)从间日疟原虫患者血样中扩增出间日疟原虫SSUrDNA片段，以PUC19质粒T载体构建重组子导入大肠杆菌JM109；阳性克隆双酶切鉴定后，双脱氧末端终止法测定序列。结果 间日疟原虫SSUrDNA扩增片段大小为341bp；阳性克隆双酶切及PCR扩增均得到预期大小的片段；序列测定插入片段为341bp，与Sal I株顺序相比，仅在第151位处缺失一个碱基C。结论 成功克隆了间日疟原虫SSUrDNA片段．该序列在间日疟原虫虫株间高度保守。     

外加磁场对半乳糖化白蛋白磁性阿霉素纳米粒在大鼠体内分布影响的研究

目的观察半乳糖化白蛋白磁性阿霉素纳米粒(ADR-GHMN)在正常肝脏中的靶向性,并观察ADR-GHMN在全身各脏器的分布特征及外加磁场对其分布的影响.方法大鼠正中开腹,肝动脉插管并固定,肝动脉注射125I-ADR-GHMN(相当于阿霉素0.5 mg/kg),左外叶加磁场,磁场应用30 min,移去磁场后,动物立即处死;对照组:肝动脉注射ADR-GHMN,左外叶不加磁场,30min后,移去磁场后,动物立即处死,立即取靶区肝、非靶区肝、肾、心、肺、小肠、脾及周围血作γ计数.肝组织作病理切片.结果注入的纳米粒75～85%分布于肝脏,其它脏器极少.病理切片显示磁区小动脉见大量纳米粒存在,对照组及非磁区肝中纳米粒很少见.结论ADR-GHMN在正常肝组织中有明显的磁靶向性;在磁场作用下,ADR-GHMN主要分布于肝脏,其它脏器含量很少;试验组肾、心、肺、小肠、脾及外周血于对照组的放射活性比较明显降低,表明磁性物质的存在使这些脏器的相对药物暴露明显减少.     

酪氨酸激酶Syk在肝癌中的表达及与血管生成的关系

目的探讨酪氨酸激酶Syk在肝细胞性肝癌中的表达及对肿瘤微血管生成的影响。方法采用RT-PCR检测Syk mRNA在肝细胞性肝癌及癌旁正常组织中的表达，免疫组化SABC法检测标本中CD34的表达反映肿瘤的微血管密度（MVD）。结果24例癌旁正常组织中Syk mRNA均表达阳性，32例肝细胞性肝癌中Syk mRNA表达率为46.9％（15／32），其中低分化组阳性表达率23．1％（3／13），明显低于高分化组阳性表达率63．2％（12／19）（P〈0．05）。肿瘤微血管密度（MVD）检测：低分化组（Ⅲ级Ⅳ级为49．2±3.6，54±4.3），明显高于高分化组（Ⅰ级Ⅱ级为13．6±4．5，32.3±3．2）与正常组织（5．9±1．7），有显著统计学意义（P〈0．05）。Syk mRNA的表达与CD34的表达明显负相关（r=-0.97）。结论肝细胞性肝癌中Syk基因的缺失对癌组织血管的生成起重要的作用。     

uPA、uPAR和PTEN在膀胱移行细胞癌中的表达及其与肿瘤侵袭和转移的关系

目的研究膀胱移行细胞癌uPA、uPAR及抑癌基因PTEN蛋白的表达及其与肿瘤侵袭、转移的关系。方法应用免疫组织化学SP法检测60例膀胱移行细胞癌组织uPA、uPAR和PTEN蛋白的表达。结果uPA、uPAR和PTEN阳性表达率分别为50.00%、50.00%和41.67%。uPA和uPAR的高表达与膀胱移行细胞癌的分化程度、浸润和转移关系密切(P<0.05);PTEN蛋白的低表达与膀胱移行细胞癌的分化程度、浸润和转移密切相关(P<0.05);膀胱癌中uPA和uP-AR与PTEN蛋白表达呈负相关(r=-0.481,P<0.05)。结论uPA和uPAR的高表达可能作为膀胱移行细胞癌侵袭、转移的指征之一,PTEN可能作为膀胱移行细胞癌预后不良的指标;uPA和uPAR对肿瘤细胞侵袭转移的作用可能在一定程度上受到PTEN的调控。     

Therapeutic effect of 5-FC on YCD modified murine P388 leukemia in vivo

Cytosine deaminases (CDs) in bacteria andfungi are found to deaminate prodrug 5 - FC intocytotoxic agent 5 - FU .Yeast CD (YCD) is clonedfrom Saccharomyces cerevisiae.It has been shownpreviously that YCD was more efficient inconversing5 - FC than bacterial CD(b CD) [1-3 ] .As anovel suicide gene therapy system,YCD/ 5 - Fcsystem may be promising in cancer therapy andprevention of graft versus host disease.In thepresent study,we established a P388/ DBA murineleukemia model by infusin…     

Determination of phospholipidosis potential based on gene expression analysis in HepG2 cells.

Phospholipidosis (PLD) is characterized by an intracellular accumulation of phospholipids in lysosomes and the concurrent development of concentric lamellar bodies. Recently, H. Sawada et al. (2005, Toxicol. Sci. 83, 282-292) identified 17 genes as potential biomarkers of PLD in HepG2 cells. The present study was undertaken to determine if this set of genes measured by quantitative PCR could be validated in the same cell line. The objective was also to investigate the dose-response relationship to further validate the assay and to select the concentrations to use for screening activities. In a first experiment (one concentration tested), out of the 17 genes, the best gene biomarkers of PLD (i.e., 11 genes) were selected for practical screening reasons. Based on these genes, 91.6% (i.e., 11 of 12) of the compounds known to induce PLD were identified as positive and all the negative compounds (i.e., five of five) were also confirmed. When the data obtained in the first experiment were compared to the data by Sawada et al., (2005) the coefficient of correlation calculated was slightly higher than 75%. In the second experiment (26 compounds [all 17 compounds from the first experiment plus 9 other compounds] tested at a minimum of three concentrations), 93.3% (14/15) of the compounds known to induce PLD were identified as such and all the negative controls (six compounds) were also confirmed. Three compounds likely to induce PLD were identified as positive in our assay. Finally, two compounds for which no data are available were also tested. When both experiments 1 and 2 were compared, the coefficient of correlation for 16 compounds tested at the same concentrations reached 87.7%. In conclusion, the present study further confirms the utility of gene expression in HepG2 cells to identify a potential to induce PLD. Finally, based on the data presented, researchers are encouraged to use a range of minimum three concentrations (e.g., 12.5, 25, and 50 microM) to screen for PLD in the human HepG2 cell line.