正交试验法优化普抗合剂制备工艺

以干浸膏得率和黄芩苷提取率为考核指标,采用正交试验法对普抗合剂水提取醇沉淀制备工艺进行考察.普抗合剂最佳制备工艺方案为加水量10倍,提取2次,每次1 h,55%的乙醇沉淀杂质 .该工艺科学合理,适合于大规模工业生产.     

猪肌肉生长抑制素基因的克隆及原核表达

本试验旨在通过基因工程方法获得重组肌肉生长抑制素蛋白。根据基因库肌肉生长抑制素基因序列设计合成特异性引物,引物两端分别加上EcoRⅠ和SalⅠ酶切位点及保护碱基,提取汉普夏猪的骨骼肌总RNA,用RT-PCR方法扩增肌肉生长抑制素全长基因,并将其克隆到pMD18-T载体上,测序后EcoRⅠ和SalⅠ双酶切,克隆到pET-30a( )中,构建原核表达载体pET-30a-M,将重组表达质粒转化至E.coliBL21(DE3)中,诱导表达。结果表明,成功扩增出肌肉生长抑制素全长基因;克隆载体经过DNA序列测定,所得基因与国外报道的完全一致;成功构建原核表达载体pET-30a-M;经IPTG诱导,表达出了6×His-M融合蛋白,用组氨酸标签抗体做Western印记证明产物大约48 ku,与预期大小相符,肌肉生长抑制素蛋白在大肠杆菌中成功的进行了表达。     

用"母犊素"提高产母犊率的应用试验

应用“母犊素“分别对荷斯坦牛、西门塔尔牛、杂种牛和土种牛提高产母犊率的试验,结果表明试验组牛平均产母犊率为763%,对照组为38.4%,比对照组提高37.9%,差异极显著(p＜0.01).品种间的差异不明显,产母犊率最高的为杂种牛达81.0%,取得了较为理想的效果.     

渭北旱塬优质苹果种植的农业地质环境特征

渭北旱塬是我国苹果种植的最优气候区。以白水县为例，通过对其气候适生区内果园地质条件的对比分析，寻找出了具体地质要素间的差异，归结出了两类气候区内地貌、土壤母质类型、母质矿物质含量三类要素的组合类型，为进一步探讨地质条件对苹果品质的影响特征，细分气候优生区，促进该区苹果生产优质化提供依据。     

科尔沁沙地农村生活能源利用与沙漠化

科尔沁沙地沙漠化的形成离不开脆弱的自然条件,与不合理的人类活动也密不可分,且与后者的关系日渐密切。针对沙漠化产生的过耕、过牧、过樵等不合理人类活动,人们采取了不同的应对措施,退耕、以草定畜、严禁采伐等。文章通过对典型区的调查和分析发现,针对过度樵采所采取的措施并不全面和彻底,长期以来,人们忽视了在半干旱气候条件下半农半牧区生物质能的合理利用,使之成为沙漠化发展的重要促进因素,需要在沙漠化防治过程中引起足够重视。     

Prokaryotic Expression and Polyclonal Antibody Preparation of σC Protein of Novel Duck Reovirus DH13 Strain

The aims of the experiment was to optimize the prokaryotic expression system of σC protein,prepare polyclonal antibody against σC protein of novel duck reovirus (NDRV),and evaluate the titer of the antibody.The σC gene of NDRV-DH13 strain was amplified by RT-PCR,ligated into pET-30a(+) and pET-32a(+) expression vector,constructed prokaryotic expression plasmid,which were transformed into E.coli BL21(DE3) and the expression of the σC protein were induced by IPTG.The proteins expression were analyzed by SDS-PAGE.The recombinant protein without His tag was purified by digestion,and the recombinant protein with His-tagged was purified by Ni-NTA column.Then the polyclonal antibody was obtained from rabbits which had been immunized by the purified protein without His tag.Anti-His-labeled mAb and NDRV-σC positive serum were used as primary antibodies to evaluate antibodies specificity,the antibodies titer was detected by indirect ELISA (iELISA).SDS-PAGE results showed that the molecular weight of the expression on recombinant proteins were 34 and 37 ku respectively,the proteins were highly expression.Western blotting showed that they had the specific reaction and the prepared antibodies had higher affinity with σC protein,the titer were about 1:25 600 by iELISA detection.This study successfully constructed and optimized the prokaryotic expression system of the polyclonal antibody against σC protein,laid a foundation for the further study of σC protein and the research of genetically engineered vaccine.     

传染性法氏囊病超强毒Gx株VP3基因在病毒致弱过程中的变异研究

利用SPF鸡胚对鸡传染性法氏囊病超强毒vvIBDV Gx株进行培育，通过鸡胚成纤维细胞对病毒进行传代致弱，对其结构蛋白VP3基因核苷酸及推导的氨基酸序列进行分析，从而揭示vvIBDV Gx从超强毒力向弱毒力转化过程中基因的序列变化规律。对不同代次细胞毒序列分析的结果表明，细胞毒在第8代以前，VP3基因序列没有改变，与标准超强毒HK46株氨基酸同源性达99％以上；细胞毒第9代核苷酸和氨基酸序列发生了改变；10代毒VP3基因与标准弱毒P2株氨基酸序列同源性达100％；细胞适应毒传至20代，其VP3基因序列不再改变。从而说明，vvIBDV Gx株在致弱过程中，VP3基因也随之改变。     

穴播条件下株行距对紫花苜蓿种子产量和质量的影响

在甘肃省酒泉市研究了穴播条件下不同株、行距处理对紫花苜蓿种子生产性能的影响，结果表明：在产种第一年（2004年）行距和株距对种子产量和产量构成因素均产生了显著的影响（P〈0．05），随着行距从60cm递增至100cm，种子产量也随之从1240kg／hm^2下降到982kg／hm^2，而各株距处理的种子产量则从15cm的1426kg／hm^2依次递减至60cm的807kg／hm^2；行距60cm和株距15cm组合的产量最高，为1833kg／hm^2，100cm行距和株距60cm组合的产量最低，为753kg／hm^2。株行距的增加显著降低了枝条密度，但结荚花序数／枝条和荚果数／结荚花序却显著提高。品种间种子产量的差异不显著。品种间的结荚花序数／枝条、种子重量、正常种苗率、发芽势和硬实率存在显著的差异（P〈0．05）。     

特异抗体致敏的SPA茵体浓缩新城疫病毒方法在临床诊断中的应用

为提高新城疫临床诊断的准确率,建立了新城疫病毒浓缩方法,将粪便及脑、肺等器官组织病料中的新城疫病毒经特异性抗体致敏的金黄色葡萄球菌(SPA菌)吸附浓缩后,用RT-PCR进行检测.结果表明,所建立的新城疫病毒浓缩方法应用于临床诊断后,可显著提高检测敏感性,从而提高临床诊断的准确率.     

Biological characters of mesenchymal stem cells of human umbilical cord blood

AIM：To study the isolation,expansion and purification of mesenchymal stem cells (MSCs) from human umbilical cord blood (UCB),and investigate some biological identities of MSCs.METHODS：(1) MSCs of UCB,adult bone marrow (BM) and fetus BM were isolated by centrifugation with Ficoll,and the different kinds of MSCs were observed everyday.(2) Surface markers of MSCs were identified by flow cytometry.(3) The level of HGFs (TPO,SCF,FLT-3L,IL-6) secreted by different sources of MSCs was checked by ELISA method.RESULTS：(1) No difference in morphology of the colonies between UCB MSCs and BM MSCs was observed.However,the mononuclear cells needed in culture of UCB MSCs was about 3 times more than that in culture of BM MSCs.The times of UCB MSCs colony formation and confluencing were longer than that of BM in primary culture.(2) After passaged,there was no significant difference in the proliferation rates of 3 kinds of MSCs.Only 4 of 15 UCB samples contained a homogeneous population of MSCs.(3) UCB MSCs shared the same markers with BM MSCs.Neither hematopoietic marker nor immunologic recognition antigens were expressed.(4) The level of hematopoietic growth factors (HGFs) secreted by 3 kinds of MSCs was similar.CONCLUSIONS：(1) MSCs were isolated from UCB,but the amount of MSCs in UCB was smaller than that in BM,and just seldom samples of UCB contained homogeneous MSCs.(2) MSCs from UCB and BM shared the same biological characteristics,such as proliferation ability,surface markers,immunophenotypes and HGFs secretion.