自动进排料密闭式固态发酵罐的设计

为解决当前固态发酵劳动强度大,生产效率低的问题,设计内置纵向排列带扇形缺口的浅盘组和升降旋转控制刮板的中心轴来实现自动进排料和翻料的固态发酵罐,可实现自动化生产,原位灭菌,提高生产效率。     

In vivo characterization of inflammatory biomarkers in swine and the impact of flunixin meglumine administration

Non-steroidal anti-inflammatory drugs (NSAID) are a family of chemicals that function to reduce pain, fever, and inflammation, and they are commonly used in people and animals for this purpose. Currently there are no NSAIDs approved for the management of inflammation in swine due to a lack of validated animal models and suitable biomarkers to assess efficacy. A previous in vitro study examining biomarkers of inflammation identified fourteen genes that were significantly altered in response to Escherichia coli lipopolysaccharide (LPS)-induced inflammation. In the present study, five of those fourteen genes were tested in vivo to determine if the same effects observed in vitro were also observed in vivo. Plasma levels of prostaglandin E(2) (PGE(2)), an essential mediator of fever and inflammation, were also determined. Two groups of swine were stimulated with LPS with the second group also treated with flunixin meglumine. Blood was collected at 0, 1, 3, 6, 8, 24, and 48h post LPS-stimulation. The RNA was extracted from the blood and quantitative real-time-PCR (qRT-PCR) was utilized to determine the expression patterns of CD1, CD4, serum amyloid A2 (SAA2), Caspase 1, and monocyte chemoattractant protein 1 (MCP-1). The LPS-stimulated animals demonstrated a statistically significant alteration in expression of SAA2 and CD1 at 3h post-stimulation. Flunixin meglumine treated animals' demonstrated reduced expression of CD1 in comparison to the LPS-stimulated swine at 24 and 48h post LPS-stimulation. Flunixin meglumine treated animals exhibited reduced expression of SAA2 at 48h post-stimulation compared to LPS-stimulated swine. Swine treated with LPS demonstrated statistically significant increases in plasma PGE(2) at 1h post-stimulation. Swine treated with flunixin meglumine had no increase in plasma PGE(2) levels at any time. These results demonstrate that PGE(2) production, along with two out of five genes (SAA2 and CD1) have the potential to serve as early biomarkers of inflammation as well as indicators of NSAID efficacy.     

牛血清中口蹄疫病毒非结构蛋白抗体时间分辨荧光免疫分析检测方法的建立

为研究快速定量检测牛血清中口蹄疫病毒非结构蛋白抗体的方法,试验通过优化抗原表达条件等步骤,在大肠杆菌原核表达系统中表达可溶性的3A-3B融合蛋白,并基于纯化的可溶性融合蛋白建立口蹄疫病毒非结构蛋白抗体时间分辨荧光免疫分析检测试剂盒。结果表明:建立的方法能够检测牛血清中的口蹄疫病毒非结构蛋白抗体,敏感性高,特异性强,对其他相关的牛类病原无交叉反应,其组内与组间变异系数分别低于10%和15%,具有良好的重复性。对300份临床牛血清样品进行检测,同Procheck公司的口蹄疫非结构蛋白抗体试剂盒进行比较,阳性样品符合率96%,阴性样品符合率93.3%,总的符合率95.7%。重复性试验组内与组间变异系数均小于10%。文章首次建立了口蹄疫病毒非结构蛋白抗体时间分辨荧光免疫分析检测方法,同传统的ELISA方法相比,该检测方法特异性相当、敏感性更高,操作更简单、快速,具有较高的应用推广价值。     

牛初乳粉片混合工艺的优化研究

为提高牛初乳粉片的混合工艺的混合均匀度,采用响应面试验设计优化混合工艺参数。在单因素试验的基础上,确定采用投料方式Ⅲ,以混合时间、装载系数为主要影响因素,以混合均匀度为响应值,采用响应面法对混合工艺参数进行优化。结果表明:牛初乳粉片混合工艺最优参数为:混合时间15 min,装载系数35%,混合均匀度为94.68%,与模型预测的理论值96.52%均比较接近。采用响应面分析优化得到的牛初乳粉片混合工艺参数具有可行性和可靠性,可用于实际操作。     

Localisation and expression of TRPV6 in all intestinal segments and kidney of laying hens

1. The aim of this study was to investigate the localisation and expression of the epithelial Ca2+ channel TRPV6 (transient receptor potential vanilloid channel type 6) in different intestinal segments and kidney of laying hens during peak lay. 2. Immunohistochemical analysis of the intestine indicated that TRPV6 was localised to the brush-border membranes of the duodenum, jejunum, ileum, caecum, and rectum. Expression was weaker in the rectum, and little or no expression was found in crypt and goblet cells. In addition, TRPV6 mRNA was quantified amongst different intestinal segments, and expression was highest in the duodenum and jejunum. Furthermore, Western blotting indicated that the duodenum expressed the greatest amount of TRPV6 and the rectum the least with the other segments expressing intermediate levels. 3. In the kidney, distinct immunopositive staining for TRPV6 was detected at the apical domain of the distal convoluted tubules (DCT) and medullary connecting tubules (CNT). Interestingly, distribution of TRPV6 extended to the proximal convoluted tubules (PCT). Furthermore, the kidney expressed lower TRPV6 mRNA and protein levels compared with that in the duodenum. 4. In conclusion, the epithelial Ca2+ channel TRPV6 is strongly expressed in the apical cells of the entire intestine and the renal tubules, suggesting that active Ca2+ transcellular transport plays a crucial role in dietary calcium (re)absorption in laying hens.     

重组质粒pACYC184-hok/sok的构建及稳定性研究

以低拷贝质粒pACYC184为载体,将hok/sok基因插入到载体BamH Ⅰ位点,构建重组质粒pACYC184-hok/sok稳定系统,同时构建hok/sok基因缺失突变重组质粒pACYC184-hok/sok(M).通过对构建的重组菌进行连续传代和对不同代次的重组质粒进行SphⅠ酶切,分析hok/sok基因的引入对宿主细胞生长的影响.结果表明,重组质粒pACYC184-hok/sok在无抗生素筛选压力下连续传代,传至第135代时质粒稳定率仍是100%,且传代前后的质粒SphⅠ酶切图谱没有发生变化,DNA序列测定表明,插入的hok/sok基因没有发生任何突变.突变重组质粒pACYC184-hok/sok(M)传代前后的质粒SphⅠ酶切结果虽没有发生变化,但其相应的重组菌传至第15代,质粒几乎完全丢失.生长曲线的测定结果表明,与舍pACYC184重组菌生长曲线相似,pACYC184-hok/sok重组菌生长较快,而pACYC184-hok/sok(M)重组菌生长缓慢.以上结果表明,含hok/sok稳定系统,其稳定性明显高于无hok/sok稳定系统的质粒pACYC184以及突变质粒pACYC084-hok/sok(M).重组质粒pAqCYC184-hok/sok具有良好的结构稳定性和分离稳定性.     

羽毛角蛋白降解的研究进展

羽毛角蛋白是一种硬性蛋白,在动物饲料、复合材料、食品包装和医药制品等领域展现出良好应用前景。但因羽毛角蛋白难溶解和降解的特殊性,一般方法很难获得可溶性角蛋白,酶法相对于传统物理和化学降解法具有反应条件温和、环境污染小和降解产物稳定等优点,是具有很高实用价值的羽毛降解途径。文中重点介绍角蛋白酶对羽毛的降解作用,并概述物理、化学方法的降解原理以及角蛋白的综合利用。     

二甲苯胺噻唑神经药理作用的研究

本实验对二甲苯胺噻唑(静松灵)的镇静、镇痛、肌松、安定和抗惊厥等中枢抑制作用进行了较为系统的研究;观察了该药对大鼠自发性脑电变化的影响;对该药的作用机理作了初步探讨。结果:(1)二甲苯胺噻唑具有良好的镇静、镇痛和松肌作用,较大剂量有一定的安定作用,但安全性差;(2)对戍四氮、士的宁和苦味毒所致惊厥反应无对抗作用;(3)对神经于神经冲动的传导具有阻断效应,(4)可诱导大鼠脑电出现高幅慢波,惊醒反应消失,妥拉苏林和育亨宾对此有拮抗作用;(5)妥拉苏林和育亨宾对二甲苯胺噻唑制动大有催醒作用,且可缓解该药的中毒反应。结果提示,中枢性α_2—肾上腺素能受体参与了二甲苯胺噻唑中枢抑制作用的产生过程。     

巴马小型猪在达芬奇机器人手术系统中的麻醉管理

为探讨巴马小型猪在达芬奇机器人微创手术中的麻醉管理策略,以丙泊酚麻醉作诱导,异氟醚维持麻醉的复合麻醉方法对巴马小型猪进行全身麻醉,同时用达芬奇机器人对其分别进行心脏、肺脏、肝脏和胃的微创手术,于术前、术中及术后监测巴马小型猪动脉血气和血流动力学变化。结果各组试验猪均顺利完成手术,手术平均时间(166±34)min,麻醉时间(181±38)min,气胸(气腹)时间(122±33)min,在整个麻醉过程及苏醒过程中,各组试验猪安全平稳,未出现体动表现。由此可见,该麻醉方法可用于达芬奇机器人手术系统,且为人医临床应用中的麻醉管理奠定基础。