Aβ1-42淀粉样蛋白在大肠杆菌中的可溶性表达

本研究中用融合标签技术通过单质粒转化和双质粒转化分别促进Aβ1-42淀粉样蛋白的可溶性表达。构建重组载体pET（yd-b42）、pET（Msb-b42）、pET（Od-b42）、pAY-sls[ydb42]、pAY-sls[tydb42],并在大肠杆菌中表达,通过SDS-PAGE验证分析。标签yd、Msb、Od、tyd都能很好的促进Aβ1-42淀粉样蛋白可溶性表达,其中yd、tyd的促溶效果最好。Aβ1-42淀粉样蛋白能在大肠杆菌中可溶性表达。为阿尔茨海默病的治疗提供进一步理论基础。     

Improved porcine model for Shiga toxin‐producing Escherichia coli infection by deprivation of colostrum feeding in newborn piglets

Porcine edema disease (ED) is a toxemia caused by enteric infection with Shiga toxin 2e (Stx2e)‐producing Escherichia coli (STEC). ED occurs most frequently during the weaning period and is manifested as emaciation associated with high mortality. In our experimental infection with a specific STEC strain, we failed to cause the suppression of weight gain in piglets, which is a typical symptom of ED, in two consecutive experiments. Therefore, we examined the effects of deprivation of colostrum on the sensitivity of newborn piglets to STEC infection. Neonatal pigs were categorized into two groups: one fed artificial milk instead of colostrum in the first 24 h after birth and then returned to the care of their mother, the other breastfed by a surrogate mother until weaning. The oral challenge with 10¹¹ colony‐forming units of virulent STEC strain on days 25, 26 and 27 caused suppression of weight gain and other ED symptoms in both groups, suggesting that colostrum deprivation from piglets was effective in enhancing susceptibility to STEC. Two successive STEC infection experiments using colostrum‐deprived piglets reproduced this result, leading us to conclude that this improved ED piglet model is more sensitive to STEC infection than the previously established models.     

一株不表达K88、K99、987P粘附因子的鸽源产毒素性大肠杆菌的分离鉴定

从病鸽尸体中分离出一株大肠杆菌,通过生化试验、血清学鉴定及动物致病性试验,表明该菌是一株具有很强致病力的不表达Ｋ８８、Ｋ９９、９８７Ｐ粘附素的鸽源产毒素性大肠杆菌。     

Surveillance for highly pathogenic avian influenza in migratory shorebirds at the terminus of the East Asian-Australasian Flyway

AIM: To determine if migratory birds arriving in New Zealand in the Southern Hemisphere spring of 2004 were infected with the highly pathogenic avian influenza (AI) virus, H5N1.

METHODS: Cloacal and faecal samples were collected from migratory red knots following their arrival in New Zealand in October 2004. Two species of resident sympatric birds, wrybill and mallard duck, were sampled prior to, and following, the arrival of migratory birds.

RESULTS: No AI viruses were isolated from migratory or resident shorebirds. Non-pathogenic AI viruses were isolated from six resident mallard ducks, comprising the endemic subtypes H4 (n=2), H7 (non-pathogenic), H10, and H11 (n=2).

CONCLUSIONS: Highly pathogenic AI H5N1 virus was not detected in migratory shorebirds or sympatric water birds in the Firth of Thames, New Zealand, in 2004-2005, despite the possible proximity of migratory birds to outbreaks of the disease in East Asia in 2004.     

猪CTSD全长cDNA的克隆和表达分析

以人CTSD mRNA全长序列为基序,在db EST库中搜索同源性大于80%、重叠大于40个碱基的猪EST下载经Seqman软件装配后,利用RT-PCR扩增到猪CTSD基因部分cDNA序列,进一步通过RACE技术获得cDNA全长（GenBank登录号为DQ018727）,猪CTSD基因cDNA全长2032 bp包括93 bp 5′UTR区、706 bp 3′UTR区和1 233bp ORF,编码410个氨基酸残基。其编码区与人、鼠、牛、羊CTSD编码区同源性分别为87.9%、78.2%、86.6%和85.0%,其氨基酸序列与人、鼠、牛、羊氨基酸序列同源性分别为86.6%、80.1%、86.0%和84.7%。疏水性分析和信号肽预测发现猪CTSD氨基酸序列存在至少7个疏水性区域,信号肽位点为第1-20个氨基酸残基。在NCBI进行Blast P搜索结果显示猪CTSD氨基酸结构中含有Asn（Asparagine,天门冬酰胺）结构域,与天冬氨酸蛋白酶3D结构同源性高达99.7%,具有真核生物天冬氨酸蛋白酶的典型结构。以猪多组织RNA池为模板,以1026 bp部分cDNA序列作为杂交探针进行Northern杂交,得到一条2000 bp左右特异条带,从而验证了RACE产物为全长cDNA并揭示该基因只有一个转录本。为了研究猪CTSD基因在体内不同组织内表达的特点,采用半定量RT-PCR对CTSD基因在猪10个组织中的表达进行研究,结果表明在10个组织中均有表达,且表达量与-βactin内参接近。     

动物大肠杆菌耐药性的变化趋势

本试验对50年代至90年代初分离保存的50株大肠杆菌进行了药物敏感性测定。结果表明，随着时间的推移，菌株对15种抗菌药物的耐性呈现不同程度的上升趋势，50年代的大肠杆菌分离株对15种抗菌药物均敏感，60年代的分离株对链霉素、四环素产生耐药，70年代对氨苄西林、氯霉素、磺胺甲基异唑、四环素、链霉素、甲氧苄胺嘧啶6种抗生素产生耐药，80年代至90年代初对阿莫西林／奥格门丁、庆大霉素、卡那霉素、萘啶酸、头孢噻吩、氨苄西林、氯霉素、磺胺甲基异唑、四环素、链霉素、甲氧苄胺嘧啶11种抗生素产生耐药，菌株的多重耐药性呈上升趋势。     

Isolation and characterization of antimicrobial-resistant Escherichia coli from national horse racetracks and private horse-riding courses in Korea

Limited information is available regarding horse-associated antimicrobial resistant (AR) Escherichia (E.) coli. This study was designed to evaluate the frequency and characterize the pattern of AR E. coli from healthy horse-associated samples. A total of 143 E. coli (4.6%) were isolated from 3,078 samples collected from three national racetracks and 14 private horse-riding courses in Korea. Thirty of the E. coli isolates (21%) showed antimicrobial resistance to at least one antimicrobial agent, and four of the AR E. coli (13.3%) were defined as multi-drug resistance. Most of the AR E. coli harbored AR genes corresponding to their antimicrobial resistance phenotypes. Four of the AR E. coli carried class 1 integrase gene (intI1), a gene associated with multi-drug resistance. Pulsed-field gel electrophoretic analysis showed no genetic relatedness among AR E. coli isolated from different facilities; however, cross-transmissions between horses or horses and environments were detected in two facilities. Although cross-transmission of AR E. coli in horses and their environments was generally low, our study suggests a risk of transmission of AR bacteria between horses and humans. Further studies are needed to evaluate the risk of possible transmission of horse-associated AR bacteria to human communities through horse riders and horse-care workers.     

一种改进的尖孢镰刀菌的实时荧光定量PCR检测方法

为实现荧光定量PCR对土壤病原真菌数量更为高效、灵敏的检测,本研究将液体培养的孢子悬浮液和长年耕作的水稻土制作成孢子浓度为4×10~1~4×10~6 spore/g的带菌土,采用MoBio PowerSoil@DNA Isolation Kit提取模拟带菌土壤总DNA,引入常规PCR预扩增包含qPCR目标序列的1 446bp片段,以预PCR产物为模板进行qPCR,构建荧光定量PCR标准曲线。研究结果表明:以试剂盒提取的带菌土壤总DNA为模板绘制的qPCR标准曲线相关系数r为0.985,检测下限为4×10~3 spore/g土;引入预PCR后,qPCR标准曲线相关系数r为0.974,检测下限为4×10~2 spore/g土,较未引入时提高了10倍,结合不同土壤病原真菌的特异性引物,该检测方法可为土壤病原真菌的有效定量检测提供技术支撑。     

细胞骨架在猪GV期与MⅡ期卵母细胞的分布规律

采用免疫荧光技术,在激光扫描共聚焦显微镜下观察并分析猪GV期和MⅡ期卵母细胞的细胞骨架分布。结果发现,猪新鲜GV期与MⅡ期卵母细胞的微管、微丝和染色体结构与分布,具有不同的特点和规律。在GV期卵母细胞,结构完整的微管尚未形成;染色体未发生致密化,仍存在于生发泡内;而微丝则分布于质膜下及整个胞质中。MⅡ期卵母细胞的微管,主要存在于纺锤体上,少量微管出现在第一极体周围;染色体排列于纺锤体赤道板上;微丝均匀分布于质膜下。本研究表明,微管、微丝和染色体三者间密切联系、相互作用,共同参与卵母细胞成熟与发育进程的调控。     

猪丹毒和猪肺疫混合感染综合诊治

疫病一直是生猪养殖中影响猪群健康的重要因素,受到疾病影响的猪群会给养殖户带来巨大的经济损失,严重阻碍养殖业的发展。该文将简要概述猪丹毒和猪肺疫的病理状况,分析猪丹毒和猪肺疫混合感染的发病特征和临床症状,并研究综合诊治猪丹毒和猪肺疫混合感染的方法,降低猪丹毒和猪肺疫混合感染对生猪养殖的影响。