The ex vivo expansion characteristic of endothelial progenitor cells

AIM: To explore the ex vivo expansion characteristics of the endothelial progenitor cells (EPCs). METHODS: CD34⁺ cells were selected from umbilical cord blood mononuclear cells (MNC) by MiniMACS system, expanded at the same conditions as that for total MNC, coincubation of CD34⁺ and CD34^- from the same donation for EPCs. In addition, we tested the effect of vessel endothelial growth factor (VEGF) and passage on cell differentiation, expansion kinetics and apoptosis. EPCs were determined and quantified by immunocytochemistry and flow cytometry. RESULTS: Coculture of CD34⁺ and CD34^-,total MNC led to a significant increase in the expansion of CD34⁺ cells compared with CD34 enrichment (P＜0.05). There was a trend toward decreased apoptosis in cultures when early passage was performed once the linear cord like structures appeared. There was no significant effect on apoptosis between with VEGF and without VEGF group (P＞0.05). These differentiated EPCs were stained positive for CD34⁺, von Willebrand factor (vWF), KDR, CD31 and incorporate acetylated low-density lipoprotein (LDL). CD34⁺ and AC133⁺cells accounted for 68.2%±6.3% (n=6) and 57.2%±9.8% (n=6) of attaching (AT) cells at day 7 of culture, respectively. CONCLUSIONS: Coculture of CD34⁺ and CD34^- or culture of MNC enhances ex vivo expansion of EPCs. Early passage decreases apoptosis rate, VEGF has no significant effect on ex vivo expansion of EPCs.     

Radioligand binding assay of insulin receptor in rabbit kidney during ischemic and reperfusion

AIM: To observe the change of insulin receptor in rabbit kidney with acute ischemic-reperfusion injury. METHODS: 15 Japanese white rabbits were allocated randomly into control group, ischemic-reperfusion group(IR group). IR group received clamping for 1 h followed by 2 h or 48 h of reperfusion. At 2 h or 48 h after reperfusion, glucose and insulin in serum were determined. Insulin receptor in renal tissue was analyzed by radioligand binging assay(BAD). RESULTS: The level of serum glucose increased after 2 h reperfusion in 2 groups, but in IR group the value increased much more higher than those in control groups(P＜0.05). Plasma insulin of IR group was significantly higher than that in control after 2 h reperfusion(P＜0.05). Scatchard analysis of data resulted in curvilinear profiles, indicating that there are two classes of receptors with different affinity or the presence of a single class of receptors with a negative cooperative hormone-receptor interaction. Data analyzed for a two-site model showed that the values of Bmax₁(high affinity site), Bmax₂(low affinity site) and Kd₁, Kd₂ were significantly lower than that of control (P＜0.05) after 2 h perfusion. 48 h after IR there was no difference of Bmax₁, Bmax₂, and Kd₁ between 2 groups,but Kd₂ of IR group was higher than that of control (P＜0.05). CONCLUSIONS: The results indicate that the effect of intrinsic insulin decreases in the progress of the renal ischemic-reperfusion. The resulting high serum glucose may aggravate renal injury in the progress of ischemic-reperfusion.     

3周龄断奶仔猪胰酶发育规律的研究

以 3周龄断奶仔猪为试验材料 ,研究了断奶对仔猪胰酶活性的影响。试验表明 ,3周龄断奶可以显著降低仔猪胰脏中胰淀粉酶、胰蛋白酶、胰糜蛋白酶和胰脂肪酶的活性 ,且在断奶后 2周继续维持断奶时水平。断奶降低了小肠内容物中胰淀粉酶和胰脂肪酶的活性 ,但空肠和回肠胰蛋白酶、胰糜蛋白酶的活性在断奶后并未下降。     

几种绿色饲料添加剂在肉鸡生产上的应用效果

选1日龄AA肉鸡390只,分成13组,分别添加不同种类和不同种类组合的绿色饲料添加剂。试验结果表明,益生素能显著(p<0.05)提高肉鸡的生产性能,改善肉品质。     

猪原始生殖嵴细胞(PGCs)建系因素的研究

从五指山猪(WSZP)近交系第8～13代培育群中，先后选用21头5～10月龄青年母猪，分别于授精后25～30d采集胎儿106个，进行原始生殖嵴(PGCs)细胞分离、培养等建系技术研究。以DMEM F10(1:1)为基础培养液，按添加或不添加生长因子，将培养液分为A、B、C3种，并以STO细胞作饲养层，在38℃、5.0％CO2和湿润的气相中进行培养建系。结果获得胚胎生殖嵴细胞(EG)细胞系6个细胞株，其中1个EG细胞株传至11代、2个传至5代、1个传至4代、2个传至3代冻存。并进行了AKP染色、体外分化、冷冻-解冻复苏和嵌合体制作等鉴定研究。研究发现：不同胚龄对EG细胞建系具有一定影响，不同培养液对EG细胞建系效果不同，STO细胞饲养层的质量是建株、传代、冷冻-解冻复苏的关键因素之一。EG细胞系的初步建立，为今后筛选进入种系的EG细胞系、实施体外基因操作提供了可能。     

植物育种的物理学方法

介绍了草原生态恢复中选育优良牧草种子的一些物理学方法,包括电场、磁场、物理辐射、激光、离子束和交变应力等在植物的生物学效应、诱变育种和转基因技术中的应用.     

日粮维生素E水平对后备种母鸡生长发育的影响

研究日粮维生素E对后备种母鸡体形、肌肉和生殖器官生长发育的影响。发现日粮维生素E水平对20周龄后备种母鸡的体重、体重均匀度、胸骨长度、胫长度、屠宰率、半净膛率、全净膛率、胸肌率、腿肌率、翅重率、腹脂率、冠高度和卵巢重量没有显著影响(P>0 05 ), 但高剂量维生素E日粮显著增加输卵管的长度和重量(P<0 05)。结果表明日粮高维生素E水平(180mg/kg) 可加速20周龄后备种母鸡输卵管的生长发育。     

高海拔地区鸡胚发育关键因素研究

对藏鸡(T)、饲养在北京的藏鸡(TB)、矮小隐性白(D)、农大小型蛋鸡商品代种蛋(M)、寿光鸡(S)、藏鸡×矮小隐性白(TD)、藏鸡×寿光鸡(TS)7组种蛋在西藏林芝进行孵化研究,通过阶段死亡率、出雏率、失水率数据分析了影响高海拔地区胚胎发育的关键因素。结果表明胚胎本身的高原生活力是鸡蛋高海拔孵化主要因素,其次是蛋壳传导力。     

鸡白细胞介素2原核表达载体的构建及表达产物的鉴定

将鸡白细胞介素2(IL-2)的cDNA克隆到原核表达载体pET-30a( )上，构建原核重组表达质粒pET-IL-2，并在大肠杆菌BL21(DE3)中诱导表达，37℃用IPTG诱导3.5h后，SDS-PAGE检测表明，表达蛋白以包涵体的形式存在。将诱导后的工程菌超声波裂解，离心后的沉淀用PBS洗涤5～6次，得到高纯度的目的蛋白。     

出血性大肠杆菌O157eae-stx1/2B融合基因的构建和表达

构建表达eae和stx1／2B的融合基因，克隆eae基因的C端280个氨基酸残基(Int280)基因部分，以正确地阅读框定向插入到含有stx1／2B融合基因的质粒，构建重组质粒，将其转化于BL21(DE3)，用IPTG进行诱导表达，经SDS—PAGE电泳检测，该融合蛋白获得了高效表达。薄层扫描分析表明：目的蛋白表达量占菌体总蛋白含量的50．67％。由于该融合蛋白由eae、stx1B、stx2B等三部分抗原组成，可刺激机体产生针对紧密素和StxB的抗体，在EHEC O157亚单位疫苗设计或单克隆抗体抗制备中具有重要价值。