PRRSV tGP5基因克隆及pcDNA3.1-tGP5真核表达载体的构建

根据GenBank公布的猪繁殖与呼吸综合征病毒(PRRSV)S1株ORF5基因的核苷酸序列设计一对特异性引物,用PCR扩增PRRSV GP5蛋白非中和性表位缺失的tGP5基因,将基因定向连接在真核表达载体pcDNA3.1的HindⅢ和EcoRⅠ多克隆位点之间,构建重组质粒pcDNA3.1-tGP5。转化DH5α感受态大肠埃希菌,提取质粒。重组质粒经PCR、双酶切鉴定及DNA序列测定,成功构建了pcDNA3.1-tGP5基因的重组体。     

Identification of RSVP14 and RSVP20 components by two-dimensional electrophoresis and Western-blotting.

We have already shown that RSVP14 and RSVP20, two ram seminal plasma (SP) proteins postulated to be involved in sperm capacitation and gamete interaction can protect spermatozoa against cold-shock. In this study, we use two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) for the analysis of SP proteins of Rasa Aragonesa rams, using enhanced protein solubilization in the presence of tributyl phosphine (TBP) and a polyacrylamide linear gradient gel with a narrow pH range (4-7). The image analysis of the 2D map detected 195 protein spots, with isoelectric points (pIs) ranging from 4.5 to 6.6, and molecular weight (M(r)) from 11.7 to 90.4. Staining of 2D gels with Pro-Q Emerald 300 Glycoprotein Stain revealed that most significant proteins in ram SP are glycosylated. The removing of protein N-linked oligosaccharides improved the gel resolution. 2D-PAGE analysis of the whole fraction 6 (F6) separated from ram SP by exclusion chromatography showed six main protein spots, four (a, b, c, d) in the 14 kDa and two (e, f) in the 20 kDa region. Western-blot analyses indicated that the anti-P14 antibody recognized four spots on the SP map, 4, 5, 6 and 7, that matched with spots a, b, c, d of F6 map. The anti-P20 antibody recognized spots 13 and 14 of SP map that corresponded to spots e, f of F6 map. The deduced sequences by de novo sequencing evidenced that protein spots 7 and 13 have significant similarities to BSP family, while protein spots 4 and 14 did not appear to be homologous with any reported protein in the current mammalian Proteinbank databases.     

Pharmacokinetics and pharmacodynamics of chlorpyrifos in male and female cattle after topical administration

The aim of this work was to study the pharmacokinetic behaviour and the inhibitory effect on acetylcholinesterase and butyrylcholinesterase activities of chlorpyrifos in male and female cattle after pour-on administration. Determination of cholinesterase activity in plasma and erythrocyte was carried out according to Ellman kinetic method. The mean baseline activities were 9338.39 ± 1331.61 and 13220.69 ± 2274.18 to acetylcholinesterase and 624.65 ± 39.32 and 641.68 ± 88.08 IU/L to butyrylcholinesterase in females and males, respectively. Acetylcholinesterase was the predominant form of cholinesterase analyzed, with low levels of butyrylcholinesterase. The basal acetylcholinesterase activities of the bulls were significantly greater than those of cows. The inhibitory effect of topical chlorpyrifos administration was lower on butyrylcholinesterase than on acetylcholinesterase. Chlorpyrifos peak plasma concentration (male:10.920 ± 4.18; female:12.12 ± 3.88 μg/L) were reached at 11.92 ± 9.19 and 8.17 ± 7.67 h in male and female, respectively. The values of area under curve were 185.96 ± 168.45 and 278.89 ± 270.00 μg·h/L and mean residence time were 13.95 ± 8.10 and 14.90 ± 9.80 h in male and female, respectively.     

Importance of house dust and storage mites in canine atopic dermatitis in the geographic region of Galicia, Spain

Sensitisation to mites is frequent in atopic dogs. The main mite genus involved in canine atopic dermatitis is Dermatophagoides. The importance of storage mite allergens in dogs has been controversial. The aim of this study was to evaluate the sensitisation rates against storage mites (Lepidoglyphus destructor and Tyrophagus putrescentiae) and house dust mites (Dermatophagoides farinae and D. pteronyssinus) in atopic dogs from Galicia, a highly humid and temperate region of Spain, using a FcepsilonRIalpha-based immunoglobulin E (IgE) in vitro test. The study was performed on 95 dogs suffering from atopic dermatitis and presenting detectable specific serum IgE levels: 91.6% of the dogs tested positive for storage mites, whereas sensitisation to house dust mites was detected in 87.4%. These results indicate the importance of storage mites in this specific geographic area.     

PK结肠溶胶囊对犬结肠炎的疗效评价

为评价PK结肠溶胶囊对犬结肠炎的治疗效果,探索更安全有效的治疗药物,以16只中华田园犬作为试验动物制作急性溃疡性结肠炎模型,随机平均分为4组,试验组分别口服PK结肠溶胶囊1颗/kg(正常剂量组)、2颗/kg(2倍剂量组)及柳氮磺吡啶结肠溶胶囊(阳性药物组,1颗/kg),测定给药后犬血液白细胞总数(WBC)、C-反应蛋白(CRP)参数变化,用内窥镜观察肠黏膜愈合状况。结果:2倍剂量组和阳性药物组给药后第4dWBC恢复到正常范围,正常剂量组给药后第7d未恢复到正常范围;2倍剂量组给药后第7dCRP恢复到正常范围,正常剂量组和阳性药物组未恢复到正常范围;内窥镜观察肠黏膜愈合状况疗效最好的是2倍剂量组。结论:PK结肠溶胶囊以2颗/kg剂量治疗犬结肠炎疗效明显,效果优于1颗/kg的正常剂量和柳氮磺吡啶。     

Effects of Exogenous Glycine Betaine on the Physiological Characteristics in Medicago sativa Seedlings under Low-temperature Stress北大核心CSCD

In this study,the 'Ganong 5th' alfalfa (Medicago sativa L.) was used as experimental material,and two application methods (leaf application,root application) and six exogenous betaine concentrations (0,10,20,30,40 and 50 mmol·L-1) were used to treat the seedlings of alfalfa. Plant growth and related physiological indexes were measured after 7 days of low temperature (4?) stress,and the physiological mechanism of exogenous betaine to alleviate the low temperature stress of alfalfa was explored. The results showed that exogenous GB alleviated the loss of plant height,root length,leaf dry weight,root dry weight and chlorophyll content,and increase the content of soluble sugar (SS) and free proline (Pro),and reduced the increase in leaf relative membrane permeability and MDA content. Exogenous GB increased antioxidant enzyme activities and non-enzymatic antioxidant content. And also,exogenous GB enhanced the osmotic adjustment capacity of the plant by increasing endogenous GB accumulation and BADH activity. These results indicated that exogenous GB enhanced the resistance of Medicago sativa to low-temperature stress under low temperature adjustment. The membership function analysis indicated that spraying 30 mmol·L-1 GB on the leaves and spraying 40 mmol·L-1 GB on the roots had the best effect on alleviating the low-temperature stress of alfalfa seedlings. © 2019 Ｊｏｕｒｎａｌ ｏｆ Ｒｅｆｒｉｇｅｒａｔｉｏｎ. All rights reserved.     

饲粮铁添加水平对产蛋期种鹅生产性能、繁殖性能、蛋品质及血液生理生化指标的影响

本试验旨在研究饲粮铁添加水平对产蛋期种鹅生产性能、繁殖性能、蛋品质及血液生理生化指标的影响,旨在探讨产蛋期种鹅对铁的需要量。选择34周龄体况相近的种鹅120只,随机分为6组,每组4个重复,每个重复5只(1公4母)。各组(Ⅰ~Ⅵ组)饲粮铁添加水平分别为0、20、40、60、80、100mg/kg。预试期1周,正试期10周。结果表明:1)Ⅴ组平均蛋重显著或极显著高于Ⅰ、Ⅱ、Ⅲ组(P<0.05或P<0.01)。由二次曲线拟合方程得出,饲粮铁添加水平为80.61mg/kg时平均蛋重最高。2)Ⅴ组种蛋合格率、种蛋孵化率和健雏率显著或极显著高于其他各组(P<0.05或P<0.01),Ⅴ组种蛋受精率显著或极显著高于Ⅰ、Ⅱ、Ⅲ、Ⅵ组(P<0.05或P<0.01)。由二次曲线拟合方程得出,饲粮铁添加水平为79.87mg/kg时,种蛋合格率最高;饲粮铁添加水平为84.82mg/kg时,种蛋受精率最高。3)Ⅴ组蛋黄色泽显著高于Ⅰ、Ⅱ、Ⅲ、Ⅳ组(P<0.05)。4)Ⅳ、Ⅴ组血清尿素氮和肌酐含量显著或极显著低于Ⅰ、Ⅱ、Ⅲ组(P<0.05或P<0.01),Ⅳ、Ⅴ组血清铁含量显著或极显著高于Ⅰ、Ⅱ、Ⅲ组(P<0.05或P<0.01)。5)Ⅴ组血液中红细胞数显著或极显著高于Ⅰ、Ⅱ组(P<0.05或P<0.01),Ⅴ组血液中红细胞压积、血红蛋白浓度、平均血红蛋白含量显著或极显著高于其他各组(P<0.05或P<0.01),Ⅳ、Ⅴ、Ⅵ组血液中平均红细胞体积显著或极显著高于Ⅰ、Ⅱ、Ⅲ组(P<0.05或P<0.01)。由此可见,饲粮铁添加水平对产蛋期种鹅平均蛋重、繁殖性能、蛋黄色泽及血液生理生化指标有显著影响,综合考虑,产蛋期种鹅饲粮铁添加水平为80.61~84.82mg/kg为宜。     

伊拉配套系曾祖代和祖代引进重庆后的繁殖性能测定分析

对重庆市引进的伊拉曾祖代4个系和祖代2个系开展连续3个胎次的繁殖性能测定,结果显示伊拉曾祖代C系平均窝产仔数和平均初生窝重显著高于A、B、D系(P<0.05),伊拉曾祖代D系21日龄平均窝重和35日龄平均断奶个体重显著高于A、B、C系;伊拉祖代D系平均窝产仔数、平均出生窝重、泌乳力(21日龄平均窝重)、35日龄平均断奶个体重均显著高于B系(P<0.05)。结果表明从法国引进的伊拉兔已基本适应重庆地区高温高湿的环境条件,初生窝重、初生产活仔数、断奶个体重等繁殖性能各项指标基本稳定。     

玉屏风多糖脂质体的制备及表征研究

优选玉屏风多糖脂质体的最佳工艺并对其进行表征验证。采用逆向蒸发法制备玉屏风多糖脂质体,以膜材比、脂药比和超声时间为考察因素,包封率为评价指标,采用L9(34)正交试验设计优选制备条件,以超速离心法测定包封率,体外释药试验验证,纳米颗粒分析仪和透射电镜测定其粒径大小与形态。制备玉屏风多糖脂质体的最佳工艺为膜材比8∶1,脂药比4∶1,超声时间90s,包封率88.38%,24h累积释放率73.45%,平均粒径132.65nm。制备的玉屏风多糖脂质体具有较高的包封率与良好的缓释作用,粒径和形态较均匀,重现性好。