Molecular Cloning and Expression of IL2 cDNA from the Tibet Pig

Interleukin-2 is a vital cytokine secreted by activated T lymphocytes, and plays important role in the regulation of cellular and humoral immunity of animals. In our experiment, IL2 cDNA of the Tibet Pig was first cloned by RT-PCR from ConA-stimulated lymphocytes in the blood and subcloned into pMD-18 T vector, which then was identified with endonuclease restriction. The sequencing result showed that Tibet pig IL-2 (TPIL-2) cDNA was 503 bp long (ORF was 465 bp) (Genbank accession number: AY 294018). The recombinant prokaryotic and eukaryotic expression plasmids of the cDNA were then constructed to analyse the ability to stimulate the proliferation of porcine lymphocytes in vitro. The recombinant porcine IL-2 expressed in the prokaryotic cells was found to be of 43 kDa molecular mass, which was consistent with a 17.4 kDa protein deduced from the IL-2 cDNA sequence (glutathione S-transferase molecular mass is 26 kDa); the recombinant protein in eukaryotic cells was confirmed by use of specific rabbit anti-porcine IL-2 serum in an ELISA. The bioactivity of TPIL-2 was detected through MTT colorimetry by stimulating the proliferation of pig ConA-stimulated blasts in vitro. The results indicate that the TPIL-2 significantly promoted the proliferation of ConA-stimulated blasts of pig. This confirms that IL-2 cDNA of the Tibet pig was successfully cloned and expressed in prokaryotic and eukaryotic cells, which lays the foundation for the the preparation of specific recombinant IL-2 protein and development of novel immune adjuvants to raise the immunity of pigs against various infectious pathogens and increase the immunoprotective efficacy of vaccines.     

冬小麦秸秆不同处理饲喂肉牛的效果观察

将冬小麦秸秆经微贮、氨化处理后进行饲喂肉牛增重育肥试验,对照牛喂自然干麦秸。48 头供试牛(每组16 头),试验结果为:喂微贮麦秸组头均日增重0.89kg,喂氨化麦秸组头均日增重0.84kg,对照组头均日增重0.57kg。试验组与对照组比较差异极显著(P< 0.01),两试验组差异不显著(P> 0.05)。将各组耗料(包括精料、粗料)、增重按市价进行效益分析,60 天试验期结束,微贮组平均每头获得114.00 元,氨化组平均每头获利63.13 元,对照组平均每头亏损14.36 元。冬小麦秸杆处理后饲喂肉牛效果明显。微贮法优于氨化法,成本低,易操作,饲喂效果好,在农区应大力推广。     

Dietary Supplementation with Microencapsulated Lutein Improves Yolk Color and Lutein Content in Fresh and Cooked Eggs of Laying Hens

This study was conducted to compare the efficacy of diet supplemented with non-microencapsulated lutein (NL) and microencapsulated lutein (ML) in laying hens. A total of 270 Hy-line Brown laying hens (54 weeks old) were allocated to three groups with six replicates of 15 hens and were adapted to a wheat-soybean meal basal diet for two weeks. Next, the control birds were fed the basal diet, and the test birds were fed the basal diet supplemented with 600 mg/kg NL (12 mg/kg available lutein) or 90.1 mg/kg ML (10 mg/kg available lutein) for 35 days. Supplementation of lutein did not affect the productive performance of laying hens, but improved (P<0.05) the yolk color and red/green value (a*), with eggs from the ML group displaying improved color and a* values from the 15^th day of the experimental period. The blue/yellow value (b*) for the yolk showed an increase (P<0.05) through both NL and ML supplements. The yolk color of fried and boiled eggs and a* value of the yolk in fried eggs were improved (P<0.05) only through ML supplemented diet. Both NL and ML supplements resulted in lower (P<0.05) lightness and higher (P<0.05) a* values of yolk in boiled eggs, as well as higher (P<0.05) b* values in fried and boiled eggs. Yolk lutein content in fresh, fried, and boiled eggs was increased (P<0.05) in NL and ML groups with the latter being higher. In conclusion, ML improved yolk pigmentation and lutein retention in laying hens better than NL.     

细胞外高钙对鸡肾小管上皮细胞活力和NO产生的影响

为探讨一氧化氮（NO）在高钙致鸡痛风中的作用，利用已建立的鸡原代肾小管上皮细胞的体外研究模型，经不同浓度的钙进行处理．通过MTT比色法观察了高钙对细胞活性的影响；采用硝酸还原酶法测定了不同浓度高钙处理原代肾小管上皮细胞后培养上清液中亚硝酸根／硝酸根（NO2^-／NO3^0）的含量（NO的静态氧化形式）。结果表明，分别用4．0mmol／L（试验组Ⅰ），8．0mmol／L（试验组Ⅱ），12．0mmol／L（试验组Ⅲ），16．0mmol／L（试验组Ⅳ）Ca^2＋处理细胞，各时段组的细胞活性与对照组相比均显著降低（P〈0．01），且细胞外钙浓度越高，细胞活性降低越明显。高钙处理细胞24h后．各组培养液中NO的含量较对照组升高不显著；48h后，试验Ⅳ组的NO比对照组显著升高；72h后，各试验组与对照组相比NO的含量均显著升高。这些结果说明，细胞外高钙能够直接损害肾小管上皮细胞，而NO可能在肾小管上皮细胞的损害中起一定的作用。     

在适应变革中实现平稳转型——在华西希望集团饲料经营分析会上的讲话

2004年是我国饲料行业具有转折意义的一年。经过这一年的市场利好,饲料行业出现了长达几年的严冬。据统计,截止于2006年底,全国已有5000多家中小饲料企业停产。2007年已经过去快半年了,由于受整个行情的影响,大企业的销量及利润大幅下滑,相当一部分企业出现亏损。[第一段]     

有机硒对种公牛精子活力、免疫力和血液生化指标的影响

试验选用5头成年种公牛作试验牛,采用分期试验,试验前期饲喂基础日粮,试验中期和后期添加一定量的有机硒,试验期采集种公牛的精液进行精子活力的检测,并分别采取血样进行血常规检测和免疫球蛋白G（IgG）的测定.试验结果表明：在种公牛日粮中添加一定量的有机硒能显著提高种公牛的精子活力和免疫力（P〈0.05）,但对血常规主要指标无显著的影响。     

BRCA1基因多态性及其与奶牛乳房炎抗性的相关性研究

为了研究BRCA1基因突变与荷斯坦奶牛体细胞数和体细胞评分的关系,试验通过对BRCA1基因外显子13、14进行克隆、序列比对和挖掘已有突变的方法确定该基因的多态位点,采用SNaPshot技术检测了BRCA1基因25025 T>A和46126 G>T突变位点在北京郊区荷斯坦奶牛群体中的分布,并对突变位点与体细胞数和体细胞评分进行了关联分析。结果表明,荷斯坦奶牛BRCA1基因2个位点均检测到3种基因型,其中25025 bp位点TT基因型为优势基因型,46126 bp位点GT基因型为优势基因型。25025 bp位点AA基因型个体体细胞数(P<0.05)和体细胞评分(P<0.01)都显著低于TT和TA基因型;46126 bp位点TT基因型个体体细胞数显著低于GG和GT基因型个体(P<0.05),但3种基因型个体体细胞评分无显著差异(P>0.05)。本研究结果初步表明,BRCA1基因25025和46126 bp位点可作为中国荷斯坦牛乳房炎抗性的标记辅助选择。     

松嫩平原艾蒿无性系种群根茎的年龄结构研究

将艾蒿根茎划分为不同年龄级,按长度和生物量统计其年龄结构,分析各龄根茎数量特征的变化规律.结果表明:艾蒿根茎的最大存活年龄为4龄;其根茎长度年龄结构为增长型;生物量年龄结构为稳定型;开花期根茎累积长度为6984.8 cm·m-2,根茎生物量为103.7 g·m-2;单位长度根茎的干物质贮存量随着龄级的增加而不断增大;高龄根茎具有较强的生活力;根茎长度比根茎生物量具有更大的生态可塑性.     

Construction and application of a bovine immune-endocrine cDNA microarray

A variety of commercial DNA arrays specific for humans and rodents are widely available; however, microarrays containing well-characterized genes to study pathway-specific gene expression are not as accessible for domestic animals, such as cattle, sheep and pigs. Therefore, a small-scale application-targeted bovine immune-endocrine cDNA array was developed to evaluate genetic pathways involved in the immune-endocrine axis of cattle during periods of altered homeostasis provoked by physiological or environmental stressors, such as infection, vaccination or disease. For this purpose, 167 cDNA sequences corresponding to immune, endocrine and inflammatory response genes were collected and categorized. Positive controls included 5 housekeeping genes (glyceraldehydes-3-phosphate dehydrogenase, hypoxanthine phosphoribosyltransferase, ribosomal protein L19, beta-actin, beta2-microglobulin) and bovine genomic DNA. Negative controls were a bacterial gene (Rhodococcus equi 17-kDa virulence-associated protein) and a partial sequence of the plasmid pACYC177. In addition, RNA extracted from un-stimulated, as well as superantigen (Staphylococcus aureus enterotoxin-A, S. aureus Cowan Pansorbin Cells) and mitogen-stimulated (LPS, ConA) bovine blood leukocytes was mixed, reverse transcribed and PCR amplified using gene-specific primers. The endocrine-associated genes were amplified from cDNA derived from un-stimulated bovine hypothalamus, pituitary, adrenal and thyroid gland tissues. The array was constructed in 4 repeating grids of 180 duplicated spots by coupling the PCR amplified 213-630 bp gene fragments onto poly-l-lysine coated glass slides. The bovine immune-endocrine arrays were standardized and preliminary gene expression profiles generated using Cy3 and Cy5 labelled cDNA from un-stimulated and ConA (5 microg/ml) stimulated PBMC of 4 healthy Holstein cows (2-4 replicate arrays/cow) in a time course study. Mononuclear cell-derived cytokine and chemokine (IL-2, IL-1alpha, TNFalpha, IFN-gamma, TGFbeta-1, MCP-1, MCP-2 and MIP-3alpha) mRNA exhibited a repeatable and consistently low expression in un-stimulated cells and at least a two-fold increased expression following 6 and 24 h ConA stimulation as compared to 0 h un-stimulated controls. In contrast, expression of antigen presenting molecules, MHC-DR, MHC-DQ and MHC-DY, were consistently at least two-fold lower following 6 and 24 h ConA stimulation. The only endocrine gene with differential expression following ConA stimulation was prolactin. Additionally, due to the high level of genetic homology between ovine, swine and bovine genes, RNA similarly acquired from sheep and pigs was evaluated and similar gene expression patterns were noted. These data demonstrate that this application-targeted array containing a set of well characterized genes can be used to determine the relative gene expression corresponding to immune-endocrine responses of cattle and related species, sheep and pigs.     

NDV基因型与其毒力、现有疫苗免疫保护之间的关系

新城疫(Newcastle Disease,ND)是由新城疫病毒(NDV)引起的侵害禽类的急性接触性传染病,该病自1926年首次在英格兰新城(Newcastle)和印尼