生长因子EGF、bFGF对猪孤雌胚体外发育的影响

在胚胎发育的不同阶段，分别在培养基中添加EGF和bFGF，研究EGF和bFGF对猪孤雌胚体外发育的作用。结果表明：在1细胞阶段添加EGF或bFGF，添加EGF能够显著提高孤雌胚的卵裂率（P〈0．05）；2～4细胞阶段添加EGF和bFGF，添加EGF组和添加bFGF组的囊胚率都显著高于对照组（P〈0．05），而添加bFGF组的囊胚率和囊胚细胞数都略高于对照组和添加EGF组。说明EGF和bFGF有利于猪孤雌胚的体外发育，而且，bF-GF能够通过提高囊胚细胞数而提高猪孤雌胚的质量。     

绿头野鸭与我国主要地方鸭品种遗传多样性研究

利用17个微卫星标记,采用PCR扩增,8%非变性聚丙烯酰胺凝胶电泳、银染,对绿头野鸭种群和我国9个主要地方鸭品种的遗传多样性进行了检测,统计了各群体的平均有效等位基因(Ne)、平均遗传杂合度(H)、多态信息含量(PIC)、群体间的遗传距离(Ds)和相似系数(I)。采用UPGMA和NJ法构建系统发生树。结果表明:17个微卫星座位在绿头野鸭种群和我国9个主要地方鸭品种中的多态信息含量均为高度多态,所有鸭种的平均PIC、H、Ne分别为0.6506、0.6896、4.1±1.5,基因多态性和遗传多样性相对较高;家鸭各品种间的遗传距离在0.1631~0.6844范围内,绿头野鸭与家鸭品种间的遗传距离分别在0.3096~0.8219之间。根据系统发育树推断中国主要地方鸭品种不是单一起源于绿头野鸭。中国主要地方鸭品种大致可以分为5支。一支为北京鸭、荆江鸭,一支为中山鸭、建昌鸭及高邮鸭,一支为靖西大麻鸭和金定鸭,连城鸭为一支,绍兴鸭为另一支。     

禽传染性支气管炎DNA疫苗在鸡体内的分布和安全性

采用本实验室构建的3种禽传染性支气管炎病毒基因的真核表达质粒pIBVS1、pIBVM、pIBVN,按各50 mg/L配制成质量浓度为150 mg/L的DNA疫苗,经腿部肌肉分点注射免疫1周龄SPF雏鸡。分别在免疫后24 h,73、0及60d,采集试验鸡的血液、心、肝、脾、肺、肾、胸腺、性腺（卵巢/睾丸）及注射部位肌肉进行组织总DNA抽提,以组织总DNA为模板进行PCR扩增。另外,在对照组DNA模板中加入不同拷贝数的质粒,确定PCR反应的灵敏性。以纯化后的组织总DNA为模板,PCR法检测质粒DNA整合到鸡细胞染色体基因组上的可能性,评价疫苗的安全性。结果表明,该疫苗在24 h内迅速分布于全身,并能在血液及所有检测的组织器官内持续分布60 d;纯化后的组织基因组DNA经PCR扩增均呈阴性,未发现整合现象,证实该DNA疫苗的安全性好。     

提高益生素作用效果的研究进展

益生素具有无污染、无毒副、无药残等特点,是抗生素的良好替代品。但益生素作用效果还存在许多争议,加强其作用效果的研究迫在眉睫。目前,人们主要通过高效菌株的选育、复合益生素的开发与益生素有协同作用的饲料添加剂研究来提高益生素的功效。寻求与益生素有协同作用的饲料添加剂被认为是最有效的方法,而广泛的应用于生产实践中。     

不同光增方式和周期对蛋鸡生产性能和血液生化指标的影响

研究光增方式和光照周期对蛋鸡蛋品质和血液生化指标的影响.将20周龄海兰褐商品蛋鸡320只,随机分为8组,每组4个重复,每个重复10只鸡.试验光照周期采用渐增和骤增2种,光周期设11L13D、13L11D、15L9D、17L7D 4个水平.结果表明①不同光增方式对蛋形指数和哈氏单位有显著影响(P＜0.05),以渐增条件下较好.不同光增方式对血液生化指标没有显著影响(P＞0.05).②不同光照周期对蛋黄颜色有显著影响(P＜0.05),以15L9D处理组蛋黄颜色最好.不同光照周期对血液生化指标有一定影响,以13L11D处理组较好.     

Classification of Morchella species from Yunnan and Zhejiang Provinces Based on rDNA ITS Sequences

Fourteen Morchella samples (eleven from Yunnan and three from Zhejiang Provinces) were selected on the basis of differences in fruit body morphology. Ribosomal DNA internal transcribed spacers (ITS) were amplified in each case using the universal primer pair, ITS-1 and ITS-4, and the amplification products were purified and sequenced. Comparisons with sequence data in GenBank revealed that the 11 Morchella isolates collected from Yunnan belonged to four species: Morchella elata, Morchella conica, Morchella crassipes and Morchella costata. The three isolates collected from Zhejiang Province (M12, M13 and M14) were designated as unknown Morchella species. When Verpa conica (AJ544206) (from the genus Verpa belonging to the same family as Morchella) was taken as the outgroup, the 14 isolates formed three groups, M. elata, M. costata (Group 1); Morchella esculenta, M. conica (Group 2); and M. crassipes, M12, M13 and M14 (Group 3).     

Construction and identification of eukaryotic expression vector of bcl-2 siRNA

AIM: To construct eukaryotic expression vector of small interfering RNA(siRNA) specific to bcl-2 and investigate the effect of recombinant plasmid on suppressing bladder cancer cell growth.METHODS: siRNA of bcl-2 gene was designed according to the principle of RNAi-based medicine, and was converted into cDNA coding expression of small hairpin RNAs(shRNA) of siRNA. The cDNA was synthesized and inserted into plasmid pGenesil-1. The recombinant eukaryotic expression vectors of pGenesil-1545 and pGenesil-1555 were controlled by the U6 promoter of RNA polymerase Ⅲ, identified by the restriction map and the sequence analysis, and transfected into T24 cells. After T24 cells were transfected for 72 h, expression of bcl-2 mRNA was assayed by RT-PCR; and MTT was used to observe the proliferation of T24 cells.RESULTS: The recombinant plasmids of pGenesil-1545 and pGenesil-1555 were identified by the restriction map and the sequence analysis. The sequences completely coincided with the designs. The expression of the bcl-2 mRNA in T24 cells transfected with recombinant plasmid decreased nearly 80%, and the growth of T24 cells was suppressed significantly.CONCLUSION: The siRNA eukaryotic expression vector against bcl-2 gene is successfully constructed. It effectively downregulates the expression of bcl-2 in T24 cells and suppresses the cell growth.     

The effects of 17β-estradiol on cardiomyocyte hypertrophy induced by endothelin

AIM:To investigate the effect of 17β-estradiol (E₂) on myocardial hypertrophy induced by endothelin-1(ET-1) and the related mechanism. METHODS:Myocardial cells from neonate rats were cultured in vitro and myocardial hypertrophy model was established with ET-1.The effects of 17β-estradiol on myocardial hypertrophy were observed. The role of ERK1/2 in the effects of 17β-estradiol was also detected. RESULTS:Compared with control group,ET-1 increased cell protein content,cell surface area and ［³H］^-Leucine ［³H］^-Leu) incorporation. Pretreatment with E₂ for 24 h could inhibit the increase in cell protein content,cell surface area and ［³H］^-Leu incorporation induced by ET-1.ET-1 significantly stimulated ERK1/2 activity,which was prevented by pretreatment with E₂.Tamoxifen,estradiol receptor antagonist,partially inhibited the effect of E₂. The ability of ET-1 to stimulate ［³H］^-Leu incorporation was significantly blocked by PD98059,which could enhance the inhibitory effect of E₂ on the increase of ［³H］^-Leu incorporation in cardiomyocytes induced by ET-1.CONCLUSION:E₂ can inhibit cardiomyocyte hypertrophy induced by ET-1. This effect is mediated by estrogen receptor. ERK1/2 signal pathway is closely correlated with the inhibitory effect of E₂ on cardiomyocyte hypertrophy induced by ET-1.     

Caspase 3 inhibitor Ac-DEVD-CHO attenuates 10-hydroxycamptothecin-induced activation of NF-κB in Bcap37 cells

AIM:To investigate the role of caspase 3 inhibitor Ac-DEVD-CHO in caspase 3 signaling pathway and NF-κB activation induced by 10-hydroxycamptothecin (HCPT) in human breast carcinoma cells. METHODS:The cell growth inhibition was measured by MTT assay. Agarose gel electrophoresis was performed for detecting cell apoptosis. Western blotting was used for determining protein expression. DIG-EMSA was conducted to measure the DNA-binding activation of NF-κB. RESULTS:Caspase 3 inhibitor Ac-DEVD-CHO attenuated HCPT-induced apoptosis in human breast carcinoma. Ac-DEVD-CHO also suppressed the degradation of caspase 3 and IκBα,and arrested the activation of NF-κB. CONCLUSION:Caspase 3 inhibitor Ac-DEVD-CHO regulates the activation of caspase 3 and NF-κB,and attenuates apoptosis in Bcap37 cell line induced by HCPT.     

应用隶属函数法综合评价葡萄种间抗寒性

在冷冻胁迫条件下,测定了葡萄属12个种45个株系枝条的膜透性变化、含水量动态变化及萌芽能力3项指标,并以种内不同株系间3项指标的平均值综合评价了12个种的抗寒性。结果表明,我国葡萄属野生种(东亚种群)山葡萄、燕山葡萄以及美洲种群中的河岸葡萄为高抗类型;蘡薁葡萄、复叶葡萄、毛葡萄为中抗类型;秋葡萄、刺葡萄、瘤枝葡萄为低抗类型;秦岭葡萄、麦黄葡萄和欧洲葡萄为不抗类型。抗寒性指标间相关性分析表明,利用平均隶属度评价葡萄抗寒性最为科学,为鉴定葡萄新种质提供了依据。