猪TLR5基因定点突变表达载体构建

TLR5特异识别细菌鞭毛蛋白,在机体免疫反应中发挥重要作用,人TLR5基因突变能影响蛋白功能并与一些疾病的易感性密切相关.研究采用RT-PCR方法克隆了猪TLR5基因的全长编码区;采用PCR介导的定点突变技术得到该基因137(G/A)和1205(C/T)位点突变的两个变异体;以pcDNA3.1+为载体成功构建野生型(TLR5-WT-pcDNA3.1+)和突变型(TLR5-G137A-pcDNA3.1+和TLR5-C1205T-pcDNA3.1+)真核表达重组质粒.研究结果可为下一步细胞水平上的突变功能分析提供基础.     

Betanodavirus ability to infect juvenile European sea bass,Dicentrarchus labrax,at different water salinity

Viral encephalopathy and retinopathy (VER) is one of the most devastating and economically relevant diseases for marine aquaculture. The presence of betanodavirus in freshwater fish is recorded, but very little is known about VER outbreaks in marine species reared in freshwater. Our study investigated the ability of betanodavirus to cause disease in European sea bass, Dicentrarchus labrax, reared at different salinity levels. Fish were challenged with RGNNV or mock infected by bath at different salinity levels (freshwater, 25‰ and 33‰). Fish were checked twice a day and the dead ones were examined by standard virological techniques, by rRT‐PCR and by histochemical and immunohistochemical analyses. All the infected groups showed a significant higher mortality rate than the one of the mock‐infected group. VERv presence was confirmed by rRT‐PCR. Histochemical and immunohistochemical analyses highlighted the typical lesions associated with VER. Our results highlight that salinity does not affect the ability of betanodavirus to induce clinical signs and mortality in European sea bass infected under experimental conditions. These results underline the great adaptation potential of VERv, which in combination with its already known high environmental resistance and broad host range, may explain the diffusion of this disease and the threat posed to aquaculture worldwide.     

SPME/GC-MS法分析不同龙眼品种果实中的香气成分

采用SPME/GC-MS法对4个龙眼品种果实香气成分进行了分析鉴定。结果表明,4个龙眼品种共检测出44种芳香物质,其中烷类11种,烯类18种,酯类10种,醇类3种,酮类1种,炔类1种,它们构成4种龙眼主要的香气成分。4个龙眼品种在香气组成和含量上有所差异,储良、东良、东丰、石硖中分别含有25种、24种、26种、21种香气成分。其中,4种龙眼共有的香气成分有7种:分别是十七烷、罗勒烯(顺式)、罗勒烯(反式)、别罗勒烯、1,3,8-对-薄荷三烯、α-石竹烯、(E)-β-金合欢烯,但其相对含量都有所差异;此外,各品种也具有自己独特的香气成分,如储良特有的香气成分有9种:包括十六烷、2-甲基-4-亚甲基-5-(2,2-二甲基环丙基)-1-戊烯、5-环丙基戊酸乙酯、二十酸乙酯、棕榈酸乙酯、9-十六碳烯酸乙酯、香叶基芳樟醇、芳樟醇、2,7-二甲基-3-辛烯-5-炔;东丰特有的3种:包括1-碘十一烷、反式,反式-法尼基酸甲酯、2-羟基十二烷酸甲酯;石硖特有的4种:包括十一烷、(-)-异丁香烯、1,3,3-三甲基-2-乙基环己烯、喇叭茶醇;东良没有特有的香气成分。     

Polyclonal activation of canine B lymphocytes evaluated by a protein A reverse hemolytic plaque assay

This paper describes the optimal culture and assay conditions for the polyclonal activation of canine lymphocytes with pokeweed mitogen and the quantitation of immunoglobulin secreting plaque-forming cells (PFC) using a staphylococcal protein A-reverse hemolytic plaque assay. The assay permits the quantitation of total immunoglobulin secreting PFC as well as class-specific immunoglobulin secreting PFC. On the optimal day of culture, a mean of 176 IgA PFC/10(6), 575 IgM PFC/10(6), 1276 IgG PFC/10(6), and 2158 total PFC/10(6) cells were generated following polyclonal activation. This study provides a simple and reproducible assay for the delineation of the immunoregulatory mechanisms involved in the differentiation of canine B lymphocytes.     

IMAGING DIAGNOSIS—MAGNETIC RESONANCE IMAGING OF DIFFUSE LEPTOMENINGEAL OLIGODENDROGLIOMATOSIS IN A DOG WITH “DURAL TAIL SIGN”

A case of diffuse leptomeningeal oligodendrogliomatosis affecting the brain and spinal cord of a dog is presented. A 7.5‐year old, male neutered Staffordshire bull terrier presented for evaluation of a chronic history of tetraparesis and seizures, with a multifocal neuroanatomical localization was determined. Extra‐axial intradural lesions with an atypical presentation of a dural tail sign were seen on MRI. Histologically, the lesions were consistent with leptomeningeal oligodendrogliomatosis. To the authors’ knowledge, a dural tail sign has not previously been reported as an MRI characteristic of diffuse leptomeningeal oligodendrogliomatosis in dogs.