口蹄疫病毒全长cDNA克隆的快速构建

利用RT PCR和 3′cDNA末端快速扩增 (RACE)技术分别扩增出包含口蹄疫病毒(FMDV)全基因组的 2个重叠cDNA片段 ,将其分别插入到载体 pcDNA3.1zeo( )和pGEM T Easy中 ,然后利用片段本身单一酶切位点将FMDV全长cDNA克隆至 pcDNA3.1zeo( )载体 ,经PCR扩增、酶切鉴定和序列测定。结果表明 ,重组质粒 (pcDNA3.1/FMDV)携带了FMDVOH/99基因组全长cDNA序列     

传染性腔上囊病病毒VP2基因表达条件的优化

用摇瓶培养法,对用作生产传染性腔上囊病基因工程亚单位疫苗的基因工程菌种BL21/pET28a VP2的表达条件进行了优化研究。其结果为:温度 37 ℃,诱导表达时间 3~7 h,用乳糖诱导时终浓度为20 mmol/L时即达到最高表达量;用IPTG诱导时终浓度为0.33 mmol/L时即达到最高表达量;乳糖的诱导效果优于IPTG。     

The Stability of Prostate-Specific Antigen in Semen Under Various Temperatures

Prostate-specific antigen (PSA) is most commonly used for identifying semen, especially in the absence of sperm. However, PSA concentration varies according to storage temperature and duration, and little is known about its stability in semen. This study was therefore aimed to determine the stability under five different temperatures: −80, −20, 4, 25, and 37°C; and nine different durations: 1, 2, 3, 5, 7, 14, 30, 90, and 180 days. All samples were stored at −80°C after being secreted from the volunteers' body until analyzed. Results showed that the PSA concentration declined significantly over time under all temperatures studied except −80°C. At −20 and 4°C, PSA was still detectable on Day 180 with 50% and 70% decrease from its original concentration, respectively. At 25 and 37°C, PSA was detected up to Day 7 and 3, respectively. This information might assist forensic scientists understand more about PSA nature and integrate it into their works.     

African biomedical scientists and the promises of “big science”

Abstract

An ongoing genomic revolution in biology has exponentially increased the rate, possibilities, scope and cost of biological research. Infectious disease genomics are often justified by the potential they have to ameliorate Africa’s disease burden. The molecular biology revolution that preceded genomic science widened the gap between the skill sets of many African biologists and their contemporaries elsewhere. Gap closure through genomics and the application of genomic data to health problems requires participation of, and leadership from, African scientists. However, few African scientists participate in genomics, and providing biological samples is their predominant contribution. Health-related applications are emerging from genomic activity for some infectious diseases that are endemic in Africa but not for many others. This article argues that the arrival of next-generation diagnostics, surveillance tools, drugs and vaccines could be accelerated by improving the nature and degree of participation of African scientists in genomic and post-genomic inquiry.     

耳穴贴压提高肿瘤危重症患者鼻肠管置管成功率临床观察

目的观察耳穴贴压法对肿瘤重症患者留置鼻肠管成功率的影响。方法将120例肿瘤危重症患者根据随机数字表法分为耳穴贴压组、甲氧氯普胺组和对照组,每组40例。统计分析3组患者置管成功率、置管时间、2次置管率、置管并发症等,监测患者置管前、置管中、置管后1 h的心率(heart rate,HR)、呼吸频率(respiratory rate,RR)、平均动脉压(mean arterial pressure,MAP)、指端血氧饱和度(percutaneous oxygen saturation,SpO 2)变化。结果耳穴贴压组、甲氧氯普胺组置管成功率均高于对照组(P<0.05),两者的二次置管率均明显低于对照组(P<0.05)。3组患者置管时间比较,差异均有统计学意义(P<0.05),甲氧氯普胺组与耳穴贴压组置管时间较对照组明显缩短(P<0.05);3组患者置管深度比较,差异无统计学意义(P>0.05)。与置管前比较,置管中3组患者HR、RR均明显加快(P<0.05),MAP明显增高(P<0.05),SpO 2均无明显变化(P>0.05);与置管中比较,置管后1 h,3组患者HR、MAP、RR均明显降低(P<0.05);与置管前比较,置管后1 h,3组患者HR、MAP、RR、SpO 2均无明显变化(P>0.05)。置管中,甲氧氯普胺组和耳穴贴压组患者HR明显低于对照组(P<0.05)。结论耳穴贴压法可提高肿瘤危重症患者鼻肠管的置管成功率,无明显并发症发生。     

鸭源新城疫强毒株对番鸭细胞因子及Toll样受体mRNA表达的影响

为研究人工感染鸭源新城疫强毒株(Md/CH/LGD/1/2005)诱导番鸭细胞因子及Toll样受体基因的表达情况,选用80只15日龄SPF番鸭,随机分为攻毒组和对照组,每组各40只,攻毒组以滴鼻点眼的方式接种1×107.0EID50鸭源基因Ⅶ型新城疫病毒(NDV)Md/CH/LGD/1/2005,对照组接种灭菌的磷酸盐...     

猪水泡病病毒非结构蛋白编码区的克隆与序列分析

以猪水泡病病毒 (Swinevesiculardiseasevirus ,SVDV)HK’70为材料 ,用特异性引物扩增出非结构蛋白编码区的 4个重叠目的片段 ,连接于pMD 18 T载体 ,转化JM 10 9感受态细胞。经对重组质粒双酶切、PCR鉴定、序列分析表明 ,HK’70非结构蛋白编码区核苷酸长 4 0 0 2nt ,氨基酸长 1334aa ;与J1’73、H/ 3’76、SVDV(Seechrn等测株 )、NET/ 1/ 92的核苷酸同源性分别为 98.6 %、98.3%、98.3%、91.2 % ,推导氨基酸序列的同源性分别为 98.9%、99.1%、99.2 %和 97.2 %。     

重组鸡白细胞介素6对不同种类疫苗的免疫增强作用

为评价重组鸡白细胞介素6(rChIL-6)在商品鸡体内对不同种类疫苗的免疫增强作用,采用不同接种方式,应用rChIL-6分别对不同种类新城疫病毒(NDV)单联疫苗和新城疫病毒-传染性支气管炎病毒-禽流感病毒H9亚型三联疫苗(NDV-IBV-H9)进行了免疫增强试验以及传染性法氏囊病病毒(IBDV)单联油乳剂灭活疫苗的免疫增强田间试验,并进行了将rChIL-6与NDV Ⅳ系预混合乳化制备成的灭活疫苗的免疫试验.结果表明,将rChIL-6与NDVⅣ系、Ⅰ系活疫苗预混合后分别采用滴鼻点眼、肌肉注射接种方式的免疫增强效果优于其他方式;rChIL-6与NDV抗原预混后制备成油乳剂灭活疫苗的免疫增强效果优于将相同抗原量的NDV油乳剂灭活疫苗和rChIL-6同时分点免疫接种.rChIL-6对NDV油乳剂灭活疫苗的免疫增强效果优于活疫苗,对NDV单联灭活疫苗的免疫增强效果优于NDV-IBV-H9三联灭活疫苗.田间试验表明,rChIL-6对IBDV油乳剂灭活疫苗首免和二免都具有显著的免疫增强作用,二免的免疫增强效果优于首免.这些研究结果为进一步开展新型鸡基因工程免疫增强剂研究奠定了基础.     

过氧化氢酶-氯霉素偶联物的合成及其结合比的测定

利用混合酸酐法合成了氯霉素琥珀酸酯衍生物．再用碳二亚胺将该衍生物与过氧化氢酶偶联,用紫外分光光度法测定已纯化的偶联物中氯霉素与过氧化氢酶的结合比。结果,偶联物中氯霉素与过氧化氢酶的结合比达到30．67:1,偶联物中过氧化氢酶的活性损失约30％。结果表明,该偶联物可作为竞争性ELISA法或免疫传感器法测试氯霉素时所需的酶标抗原。     

多重RT-PCR快速检测鉴别新城疫病毒强毒株和弱毒疫苗株方法的建立

根据新城疫病毒(NDV)基因结构的特点及强、弱毒株融合基因裂解位点的序列差异设计了4条引物,建立了一种可以快速鉴别NDV强毒株和弱毒疫苗株的多重PCR方法。检测结果显示,强毒株可以扩增出442 bp的特异性片段和671 bp的通用片段,弱毒疫苗株可以扩增出252bp的特异性片段和671 bp的通用片段。该方法只需进行一次RT-PCR,整个过程可在数小时内完成。经敏感性测定,该方法最低能检测到100 pg的NDV RNA。