四君子汤对脾虚大鼠空肠黏膜损伤的修复作用及机制

通过检测脾虚大鼠空肠中PCNA和IGF-1的表达,探讨四君子汤对脾虚大鼠空肠黏膜修复的影响。试验分对照组、脾虚组、四君子颗粒治疗组和治疗对照组,每组采取空肠中段作样本;H.E.染色法观察空肠组织结构,免疫组织化学法检测PCNA和IGF-1的表达。脾虚组大鼠空肠绒毛长度、隐窝深度极显著降低(P<0.01);四君子颗粒治疗组大鼠绒毛长度、隐窝深度的显著高于治疗对照组(P<0.05)。脾虚组大鼠空肠中PCNA、IGF-1阳性细胞数极显著增加(P<0.01);四君子颗粒治疗组PCNA、IGF-1阳性细胞个数显著高于治疗对照组(P<0.05)。结果表明,四君子汤可以促进脾虚模型大鼠空肠的修复过程。     

沙门菌CRISPR的结构与功能研究进展

近年发现沙门菌中恒定地存在两个CRISPR位点,沙门菌利用它们不仅能抵御外源质粒和噬菌体的入侵,还能介导自身短期的表型变化及长期亚类的进化。同时由于其结构的多样性,使之广泛应用于沙门菌分型和进化的研究。论文简要综述了沙门菌CRISPR系统的基本结构、作用机制和功能及其在沙门菌分型和进化应用方面的研究进展。     

Effects of Feeding Different Dietary Crude Protein and Digestible Energy to Yili Mares during Late Lactation Stage on Growth Development and Blood Biochemical Parameters of Foals

In this paper, thirty Yili mares aged 11 to 12 years with weighing 410 kg ± 30 kg, parity of 4 to 5 in third lactation month were selected, and were randomly divided into six groups, each group with five repeats.Two factors quadratic regression orthogonal rotational combination design was adopted to evaluate the effects of feeding different dietary CP and DE to mares on growth development and blood biochemical parameters of foals.Daily DE and CP feeding levels of the 6 groups were 92 MJ/d and 1.20 kg/d, 125 MJ/d and 1.20 kg/d, 92 MJ/d and 1.55 kg/d, 107 MJ/d and 1.35 kg/d, 125 MJ/d and 1.44 kg/d, 115 MJ/d and 1.55 kg/d, respectively.The results showed that body weight, body height, chest circumference and cannon circumference of foals of four and five months were not significant affected by maternal protein and energy intake (P>0.05).As compared with ration contained high DE (125 MJ/d) and low CP (1.20 kg/d), feeding with ration contained moderate DE (107 MJ/d) and CP (1.35 kg/d) improved body length of foals of four and five months significantly (P<0.05).From the blood biochemical parameters, such as total protein, albumin, triglyceride, high-density lipoprotein cholesterol, low density lipoprotein cholesterol, urea nitrogen, creatinine, alanine aminotransferase, there were no significant differences among all groups (P>0.05).Mares fed with ration contained moderate DE (107 MJ/d) and CP (1.35 kg/d) obviously improved the aspartate aminotransferase and uric acid level.In summary, Yili mares during lactation late stage fed with 1.35 kg/d CP and 107 MJ/d DE daily could significantly improve body length development of foals, and foals nitrogen metabolism was altered by maternal dietary nutrition through increasing blood aspartate aminotransferase activity.     

Identification and Molecular Characteristics Analysis of Porcine Epidemic Diarrhea Virus Variants

To ascertain a diarrhea case in a pig farm in Shandong province,the pathological changes of dead piglets were observed and nested RT-PCR test was carried out on 7 diarrhea samples for porcine epidemic diarrhea virus (PEDV).Pathological examination revealed that intestine was detected as enlargement,hyperemia and edema.After histological examination,the typical microscopic lesions of intestine were disappearance of epithelial cells,villus shrinkage and shortening.The result of nested RT-PCR showed that all of the 7 samples could amplify a specific target band of PEDV.Molecular characteristics of two field strains showed that they had an amino acid homology of 92.3% to 92.4% with vaccine CV777 and 96.6% to 98.6% with other previous field strains which sequences were downloaded from GenBank.Phylogenetic tree analysis further revealed that all of PEDV strains could be mainly divided into two clusters of G1 and G2. G2 consisted of the field strains of our study and other field strains from USA,China and so on,which had the same sequence characteristics of two insertions and one deletion,while G1 consisted of all vaccines of CV777 and several older field strains from China and Korea.These results indicated that our field strains were the dominant strains in recent epidemic diarrhea occurrence.Moreover,its molecular characteristics might be a characterization of differentiating the field and vaccine strains.     

Marc145细胞源EDC3基因的克隆及其生物信息学分析

为获得Marc145细胞源EDC3基因序列,分析其序列特征及编码蛋白的结构和功能,本试验采用RT-PCR方法从Marc145细胞中扩增EDC3基因,并进行克隆和序列测定;应用DNAStar软件分析该基因的核苷酸和氨基酸序列,与参考序列经BLAST比对后分析同源性,并构建系统进化树;利用生物信息学方法对其编码区蛋白进行二级结构、三级结构、B细胞表位、跨膜结构域和信号肽预测。结果显示,Marc145细胞源EDC3基因长度为1 527 bp,共编码507个氨基酸;EDC3基因编码区核苷酸序列与绿猴、猕猴、狒狒、人、倭黑猩猩、马、野猪、虎鲸、绵羊、非洲象和大熊猫的同源性在91.5%~99.2%之间,与非哺乳动物原鸡同源性最低,仅为81.2%;EDC3氨基酸序列与上述物种的同源性在95.3%~99.6%之间,与原鸡的同源性仅为88.8%。系统进化树结果显示,Marc145细胞源EDC3基因与绿猴的亲缘关系最近,其次是灵长类。蛋白结构预测结果表明,EDC3蛋白主要由α-螺旋和无规则卷曲组成,分别为23.38%和47.35%,二级结构与三级结构预测结果相符。该蛋白存在多个B细胞优势抗原表位,无跨膜结构域及信号肽区域。本试验结果可为Marc145细胞源EDC3基因功能的进一步研究提供参考。     

包衣技术提高药物药学性能的研究进展

兽药对畜牧生产中的疾病防控和健康养殖意义重大,但由于某些药物味苦、对胃黏膜刺激性大、稳定性差和药理作用部位靶向性差等限制了其在临床上的使用。包衣技术在改善药物适口性、提高药物稳定性和实现药物控缓释和靶向性等方面具有显著优势。作者总结了国内外关于包衣技术在改善药物适口性、提高药物稳定性、制备控缓释剂型和高效靶向释药剂型方面的研究进展,分析了包衣膜的形成原理,剖析了几种常见包衣技术的优缺点和其产业化面临的挑战,在此基础上展望了包衣技术在兽药领域的应用前景,以期为基于包衣技术的兽药新制剂的开发提供新思路。     

Preliminary Study on NF-κB Signaling Pathway Regulating Brucella Intracellular Survival Molecular Mechanisms

By the infection of Brucella virulent strain and attenuated strain in mice macrophage RAW264.7,the assay was aimed to explore the relationship between NF-κB signaling pathways and Brucella virulent strain and attenuated strain in intracellular survival.Use different MOI Brucella (2308,RB51,16M and M5) to infect mice macrophage RAW264.7,after 0,4,8 and 24 h infected,cracking cell and collecting supernatant,we detected the effect of Brucella on activation of NF-κB signaling pathway by Western blotting.Different concentrations of NF-κB signaling pathway inhibitor were incubated with mice macrophage RAW264.7,with different multiplicities of infection (MOI) of Brucella infecting cells,ELISA kits to detect the expressions of TNF-α,IL-1β and IL-6 cytokine;At the same time,count the number of intracellular bacteria of CFU.The results showed that rough cattle Brucella strains RB51 could strongly activate NF-κB signaling pathway,smooth cattle Brucella strains 2308 was weak in the activation;At the same time,the activation of NF-κB signaling pathway was concentration dependent.When the MOI was 80,infection time was 8 h,NF-κB activation degrees of rough cattle Brucella strains RB51 and smooth cattle Brucella strains 2308 were the strongest,and this pathway was involved in producing TNF-α and IL-6;NF-κB signaling pathway inhibitor BAY11-7082 affected Brucella intracellular survival.So rough cattle Brucella strains RB51 intracellular survival and NF-κB signaling pathway activity were closely related.The results laid the foundation for the further study of Brucella intracellular pathogenesis,also provided scientific basis for the research of new drugs to Brucella,and prevention and treatment of brucellosis.     

Establishment and Optimization of INH Epitope Peptide Vaccine for Detection of Antibody

In this study, an indirect ELISA method was established to detect inhibin hormone (INH) epitope peptide vaccine antibody, it would provide oretical reference for the determination of Fine-wool sheep after active immune body INH epitope peptide vaccine antibody. On the basis of the predecessors, using indirect ELISA method to determinate serum INH epitope peptide antibody levels of sheep, and through control different experimental conditions to look for the best experimental conditions. Through explorating the experimental conditions, finally, the testing experiment conditions were determined, which was blocked solution with skimmed milk powder, INH and GnIH synthetic peptides dilution degrees for 20 000 times, the optimum reaction time was 60 min, the best color action time was 15 min. In this experiment, a kind of method to detection antibody in the body after INH active immune sheep was built, it would provide a reference for future research.     

Effect of Different Concentrate-roughage Ratios on Rumen Degradability of Cassava Stems and Leaves Total Mixed Ration in Goat

The rumen degradations of TMR were studied in this experiment,in which cassava stems and leaves were used as roughage and formulated with concentrate in different proportions.The purpose of this study was to formulate the available TMR for finishing goat,and provide a theoretical basis for the applications of cassava stems and leaves in goat production.Six Hainan Black goats with permanent rumen fistulas were selected,the rumen degradabilities of DM,CP,NDF and ADF of TMR with 6 different concentrate to forage ratio（2:8,3:7,4:6,5:5,6:4 and 7:3）were tested using nylon bag method.The results showed that,the DM effective degradabilities at the ratio of 4:6,5:5,6:4,7:3 were significantly higher than that of 2:8 and 3:7（P<0.05).CP effective degradabilities at the ratio of 2:8,3:7,4:6 were significantly higher than that of 5:5,6:4 and 7:3（P<0.05).The NDF effective degradation at the ratio of 5:5 was significantly higher than that in the other groups（P<0.05).The ADF degradation at the ratio of 3:7 was significantly higher than that of 4:6,5:5, 6:4 and 7:3（P<0.05).The results indicated that,the nutrient degradation in rumen were higher when the concentrate to forage（cassava stems and leaves）ratio in TMR were 3:7,4:6 and 5:5,and the recommended ratio range were 4:6 to 5:5 in production practice.     

F107(F18ab)菌毛单抗的制备、鉴定与初步应用

将大肠杆菌107／86和水肿病菌株在TSB中培养，鉴定其表达F107菌毛，前者作为免疫原免疫BABL／C小鼠，后者作为检测抗原包装酶标板。细胞融合后经ELISA和玻板凝集检测，得到一株阳性杂交瘤细胞株，命名为F7－1。该细胞株制备的腹水ELISA效价为：2^16，试管凝集价为：1：5120，在免疫印迹中可与提取的F107菌毛特异性条带发生反应。利用此单抗检测表明，91％的水肿病菌株表达F107菌毛。