国内埃博拉病毒研究最新动态

对国内埃博拉病毒诊疗检测产品的近期研究成果及相关专利进行总结归纳,旨在为埃博拉病毒检测试剂及治疗药物的进一步开发提供参考。     

Cell Growth and Flavonoids Production in Suspension Culture of Saussurea medusa

Cell suspension cultures were established from Saussurea medusa Maxim. callus cultures. The effects of different rotation speeds of the gyratory shaker, different inoeulum sizes and different pH values of the medium on cell growth and flavonoid formation were studied. The result showed that the optimum rotation speed, inoeulum size and initial pH value of the medium were 90–120 r/min,50– 80 g FW/L and 5.5–6.0 respectively for cell growth and flavonoids formation in the suspension cultures. Sucrose was better than glucose and fructose for the suspension cultures. The optimum concentration of sucrose for cell growth and flavonoid production was 40 g/L, and the concentration of flavonoids could be as high as 1 423.25 mg/L. High performance liquid chromatographic analysis of cell suspension culture extracts showed that the concentrations of jaceosidin and hispidulin in the flavonoids were 22.11% and 0. 15% respectively.     

Prevalence and evolution of drug resistance HIV-1 variants in Henan, China

To understand the prevalence and evolution of drug resistant HIV strains in Henan China after the implementation of free antiretroviral therapy for AIDS patients. 45 drug naive AIDS patients, 118 AIDS patients who received three months antiretroviral therapy and 124 AIDS patients who received six months antiretroviral treatment were recruited in the southern part of Henan province. Information on general condition, antiretroviral medicines, adherence and clinical syndromes were collected by face to face interview. Meanwhile, 14ml EDTA anticoagulant blood was drawn. CD4/CD8 T cell count, viral load and genotypic drug resistance were tested. The rates of clinical improvement were 55.1% and 50.8% respectively three months and six months after antiretroviral therapy. The mean CD4 cell count after antiretroviral therapy was significantly higher than in drug naive patients. The prevalence rate of drug resistant HIV strains were 13.9%, 45.4% and 62.7% in drug naive patients, three month treatment patients and six month treatment patients, respectively.The number of resistance mutation codons and the frequency of mutations increased significantly with continued antiretroviral therapy. The mutation sites were primarily at the 103, 106 and 215 codons in the three-month treatment group and they increased to 15 codon mutations in the six-month treatment group. From this result, the evolution of drug resistant strains was inferred to begin with the high level NNRTI resistant strain, and then develop low level resistant strains to NRTIs. The HIV strains with high level resistance to NVP and low level resistance to AZT and DDI were highly prevalent because of the AZT DDI NVP combination therapy. These HIV strains were also cross resistant to DLV, EFV, DDC and D4T. Poor adherence to therapy was believed to be the main reason for the emergence and prevalence of drug resistant HIV strains. The prevalence of drug resistant HIV strains was increased with the continuation of antiretroviral therapy in the southern part of Henan province. Measures, that could promote high level adherence,provide new drugs and change ART regimens in failing patients, should be implemented as soon as possible.     

小粒野生稻STK类抗病基因同源序列的克隆与分析

根据丝氨酸/苏氨酸蛋白激酶类(serine-threonine kinase,STK)抗病基因结构中保守结构域,设计引物,以小粒野生稻基因组DNA为模板,扩增获得10条STK类抗病基因同源序列.同源性分析表明其都具有STK保守结构域,与已克隆的STK类抗病基因有不同程度的相似性,为进一步克隆小粒野生稻中的STK类抗病基因提供依据.     

抗人胎盘酸性铁蛋白单链抗体的构建、表达与鉴定

目的:制备人胎盘酸性铁蛋白(PAF),构建、表达并鉴定抗人PAF单链可变区抗体片段(scFv)。方法:设计以SfiⅠ、NotⅠ为酶切位点、以(Gly4Ser)3为linker的2对引物,从抗人PAF单克隆抗体可变区基因的克隆载体中扩增VH和VL基因,用重叠延伸PCR在VH和VL基因间引入连接短肽,构建VH-linker-VL的scFv基因。经SfiⅠ、NotⅠ酶切后克隆到原核分泌型表达载体pUC19/119上,转化大肠杆菌TG1和筛选后测序验证,IPTG诱导表达,SDS-PAGE鉴定其相对分子质量(Mr)。以人PAF为抗原,scFv为一抗,抗His单克隆抗体为二抗,间接ELISA鉴定其抗体活性。结果:重叠延伸PCR扩增产物经凝胶电泳可见约700bp的条带,DNA序列分析证明2株抗体具有完整的scFv序列,并含有c-myc和His。IPTG诱导阳性菌表达产物经SDS-PAGE鉴定有Mr约27000的显示条带,符合scFv与表达标签融合蛋白的理论值;间接ELISA证明表达产物可与PAF结合。结论:成功构建并表达了抗人PAF单链抗体,为进一步建立检测PAF的间接ELISA奠定了基础。     

Ca2 +对小麦种根及其根毛生长发育的影响

低浓度的CaCl2对小麦种根生长、根毛发生和生长无明显影响,高浓度的CaCl2(0.1 mol/L)对种根和根毛生长有抑制作用,但不影响根毛的发生。Ca2+专一性整合剂EGTA抑制种根生长和根毛的发生及生长,添加一定浓度的外源CaCl2,这种抑制作用可被消除。CaM抑制剂TFP(三氟拉嗪)和CPZ(氯丙嗪)对种根生长、根毛发生和生长均有抑制作用,添加外源CaM可减弱或消除这种抑制作用。     

应用RAPD方法对近交系小鼠进行遗传检测的研究

目的　为实验室日常检测近交系小鼠的遗传背景提供一种分子生物学方法。方法　用 6条随机引物对 6个品系近交系小鼠基因组DNA进行PCR扩增。结果　 6条随机引物中p2、p3、p5和p6四引物扩增的条带差异较为明显。结论　RAPD方法是一种有效的近交系小鼠遗传检测手段     

灰葡萄孢丝裂原活化蛋白激酶编码基因bmp1和bmp3的功能

【背景】植物病原真菌丝裂原活化蛋白激酶(Mitogen-activated protein kinase,MAPK)信号途径参与病菌有性生殖、细胞壁完整、菌丝侵染、致病力、胁迫响应等过程,灰葡萄孢MAPK信号途径参与病菌生长发育、致病力以及胁迫响应,但MAPK信号途径基因在灰葡萄孢中的功能尚未完全阐明,该信号途径对灰葡萄孢的生长发育和致病力的调控机制尚不明确。【目的】明确灰葡萄孢MAPK编码基因bmp1、bmp3在病菌生长发育、致病力以及氧化胁迫响应过程的功能,为进一步阐明MAPK信号途径调控灰葡萄孢生长发育和致病力的分子机制奠定基础。【方法】利用RNAi技术构建灰葡萄孢MAPK编码基因bmp1和bmp3的RNAi突变体,并以野生型BC22菌株为对照,对bmp1和bmp3基因的RNAi突变体的表型、致病力以及对氧化胁迫的敏感性进行分析。【结果】灰葡萄孢bmp1和bmp3基因的RNAi突变体其菌落形态、菌丝形态均与野生型BC22菌株没有明显差别;bmp1基因的RNAi突变体生长速率明显减慢,分生孢子产量明显降低;bmp3基因的RNAi突变体的生长速率与野生型BC22菌株没有明显差别,不能产生分生孢子。bmp1和bmp3基因的RNAi突变体在番茄果实的表面均不能产生明显的致病症状,而且不能穿透玻璃纸。bmp1基因的RNAi突变体在含有H_2O_2的培养基上受抑制的程度显著低于野生型,而在含甲萘醌的培养基上受抑制的程度显著高于野生型;bmp3基因的RNAi突变体在含有H_2O_2和甲萘醌的培养基受抑制的程度均显著高于野生型。【结论】灰葡萄孢bmp1基因正调控病菌生长、分生孢子形成、致病力和穿透能力,参与调控病菌对氧化胁迫的响应;灰葡萄孢bmp3基因正调控病菌分生孢子形成、致病力、穿透能力以及对氧化胁迫的响应。     

广东石门台自然保护区的蕨类植物区系分析

通过野外调查和标本参考,广东英德石门台国家自然保护区蕨类植物区系共含47科, 96属, 293种。分析表明:(1)热带、亚热带分布性质的科为主(31科),占66.0%,非热带性分布科(16科),占34.4%。(2)热带、亚热带性质的属占优势(64属,占66.7%),其中泛热带分布属(24,25.0%)和热带亚洲分布属(19, 19.8%)是最重要的分布类型。世界分布属占26.4%,温带分布属很不明显,占7.3%。属的这种分布格局,显示出该区蕨类植物区系从热带成分向亚热带成分的渗透和过渡。(3)热带、亚热带分布种(139种)所占比例最大(47.4%),其中以热带亚洲性质的占优势(103, 35.2%);温带成分占较大的比例(34.1%)。种的分析结果与属的分析结果相差很大,出现区系相互渗透的现象。(4)在种的相似性比较上,与东部车八岭的相似性最高(38.5/59.2);与西部大瑶山种的相似性最低(21.5/35.6)。该区的蕨类植物区系与周围地区关系较为密切。     

重金属在鲫鱼卵巢的富集及对卵细胞的损伤效应

近些年来,随着国民经济的快速发展,环境和生态问题也日益凸显,其中水域环境受重金属的污染亦愈演愈烈,水生动物的生存受到严峻的挑战。在水环境中,各种重金属元素不但在水生动物的皮肤、肌肉、鳃和其他内脏器官中富集,而且在其赖以生息繁衍、绵延种群的