对职业生涯规划教育贯穿人才培养全过程的思考

当前,职业生涯规划教育越来越受到高职院校的重视,在全民创业、万众创新的时代,更需要系统考虑职业生涯规划教育的开展与落实,使学生认识到职业生涯规划教育的重要意义。     

五种药用植物提取液抑菌作用研究

采集石菖蒲、酸浆、矮茶子、鸡屎藤和串果藤5种药用植物,以乙醇为溶剂提取其有效成分,并将提取液分别对大肠杆菌、变形杆菌和枯草芽孢杆菌进行抑菌活性试验。试验结果表明,5种植物都具有抑菌作用,但不同植物对不同细菌的抑菌效果有差异。石菖蒲和酸浆提取液对3种细菌都具有较强抑制能力,且对高温稳定。矮茶子、鸡屎藤提取液对大肠杆菌有抑制作用,串果藤提取液对枯草芽孢杆菌有抑制作用,但其抑菌能力不及石菖蒲和酸浆2种植物。     

Identification of regulated proteins by resveratrol in glutamate-induced cortical injury of newborn rats

Glutamate induces neuronal damage by generating oxidative stress and neurotoxicities. The neurological damage caused by glutamate is more severe during brain development in newborns than in adults. Resveratrol is naturally present in a variety of fruits and medicinal plants and exerts a neuroprotective effect against brain damage. The goal of this study was to evaluate the neuroprotective effects of resveratrol and to identify changed proteins in response to resveratrol treatment during glutamate-induced neonatal cortical damage. Sprague-Dawley rat pups (7 days old) were randomly divided into vehicle, resveratrol, glutamate, and glutamate and resveratrol groups. The animals were intraperitoneally injected with glutamate (10 mg/kg) and/or resveratrol (20 mg/kg) and their brain tissue was collected 4 hr after drug administration. Glutamate exposure caused severe histopathological changes, while resveratrol attenuated this damage. We identified regulated proteins by resveratrol in glutamate-induced cortical damaged tissue using two-dimensional gel electrophoresis and mass spectrometry. Among identified proteins, we focused on eukaryotic initiation factor 4A2, γ-enolase, protein phosphatase 2A subunit B, and isocitrate dehydrogenase. These proteins decreased in the glutamate-treated group, whereas the combination treatment of glutamate and resveratrol attenuated these protein reductions. These proteins are anti-oxidant proteins and anti-apoptotic proteins. These results suggest that glutamate induces brain cortical damage in newborns; resveratrol exerts a neuroprotective effect by controlling expression of various proteins with anti-oxidant and anti-apoptotic functions.     

鸡FKBP5基因的克隆、生物信息学及组织表达谱分析

本研究旨在对鸡FK506结合蛋白5(FK506 binding protein 5,FKBP5)基因进行克隆和生物信息学分析,并检测其在鸡不同组织中的表达量。以鸡脾脏cDNA为模板,通过PCR方法扩增和克隆鸡FKBP5基因完整CDS区序列,并进行同源性比对及系统进化树构建;使用在线软件分析FKBP5蛋白的理化性质、疏水性、跨膜结构、信号肽、二级结构和三级结构;利用实时荧光定量PCR检测其在肢体内外翻畸形(valgus-varus deformity,VVD)组肉鸡和正常组肉鸡组织中的表达量,并绘制组织表达谱。结果显示,鸡FKBP5基因CDS区序列全长1350 bp,编码449个氨基酸;同源性比对和进化树分析表明,鸡FKBP5基因氨基酸序列与鹌鹑、鸭、壁虎、中华鳖、小鼠、人、猪、斑马鱼的同源性分别为98.4%、96.2%、85.8%、87.4%、81.1%、85.2%、86.1%和61.2%,与鹌鹑、鸭的亲缘关系最近,爬行类和哺乳类动物次之,与斑马鱼(鱼类)亲缘关系最远。鸡FKBP5蛋白分子质量为50.43 ku,理论等电点(pI)为5.94,半衰期为30 h,肽链N端为蛋氨酸(Met),不稳定系数为23.80,属于稳定蛋白;跨膜区和信号肽预测结果显示,FKBP5蛋白不属于跨膜蛋白和分泌型蛋白。结构域分析结果表明,该蛋白包括2个FKBP型肽基脯氨酰异构酶(FKBP-type peptidylprolyl isomerases)、3个四肽重复(TPR)序列,主要包含α-螺旋(46.77%)、延伸链(12.47%)和无规则卷曲(40.76%),且该蛋白与HSPA2、PPID、STIP1、HSP90AA1和HSP90AB1等互作蛋白具有很强的相关性。实时荧光定量PCR结果显示,FKBP5基因在鸡各组织中广泛表达,其中在软骨和法氏囊中表达量较高,在发病组肉鸡组织中的表达量均高于正常组肉鸡,且在心脏、腿肌、法氏囊、胸腺、软骨中表达量差异显著(P<0.05),在脾脏中表达量差异极显著(P<0.01)。本试验结果可为肉鸡骨骼疾病及腿部健康选育提供参考。     

拷贝数变异在畜禽中的研究进展

拷贝数变异（copy number variation,CNV）具有多种形式的变异结构,在品种多样性、生物进化和疾病相关性等研究中起着重要作用,并具有片段长度大、覆盖范围广等特点。随着分子生物学的发展及DNA测序技术的日渐成熟,人们对遗传变异的研究不断向DNA分子水平深入,多态性标记在畜禽育种中已逐渐成为动物育种研究的趋势和主流。由于CNV对基因的调控和表达所造成的影响更为显著,因此,CNV在重要畜禽中的研究越来越多。目前,已检测出大量有关畜禽重要经济性状的基因序列变异,并有许多研究均表明CNV与动物的重要经济特征及疾病的发生有关。笔者主要通过参考国内外相关的研究报道,简述了CNV的相关研究背景、概念、突变机制,归纳总结了CNV对牛、羊、猪、鸡的经济性状、繁殖性状和疾病调控的影响,以期通过对这些重要畜禽的基因组学研究揭示其适应性遗传机理和表型性状差异的遗传基础,开发相应的分子遗传标记,为畜禽的标记辅助选育提供理论基础。     

猪圆环病毒2型灭活疫苗中病毒抗原含量实时荧光定量PCR检测方法的建立

依据GenBank公布的猪圆环病毒2型Cap基因序列,在保守区域设计特异性引物和TaqMan探针,优化反应体系,建立评价猪圆环病毒2型灭活疫苗中病毒含量的实时荧光定量PCR检测方法,对方法的特异性、敏感性和重复性进行试验,并验证灭活剂用量和灭活时间对检测结果的影响。结果显示：该方法只对猪圆环病毒2型基因有特异性扩增,其他3种对照病毒基因的扩增结果均为阴性;检测灵敏度达到10^2.0 TCID₅₀/mL,比普通PCR方法高100倍;方法的重复性好,对同一样品进行10次检测,变异系数为2.28%;不同灭活剂用量和灭活时间对结果的影响较小,不会因各厂家使用的灭活剂用量和灭活时间不同,影响疫苗对比实验的公平性。该研究成功建立了一种评价猪圆环病毒2型灭活疫苗中病毒含量的实时荧光定量PCR检测方法,用于猪圆环病毒2型灭活疫苗样品中病毒抗原含量的定量,其结果可以反映不同猪圆环病毒2型灭活疫苗样品中抗原含量差异,为研究猪圆环病毒2型灭活疫苗病毒抗原含量评估方法提供了新的思路。     

Prokaryotic Expression and Antigenicity Analysis of Porcine Japenese Encephalitis Virus Envelope Protein Domain Ⅲ

In order to analyze the antigenicity of porcine Japanese encephalitis virus (JEV) E protein domain Ⅲ, which was expressed by pET-28a vector with His-tag and purified through Ni-NTA, the BALB/c mice were immunized with the purified protein.We identified the antigenicity of domain Ⅲ of E protein and the anti-mice and anti-porcine JEV E Ⅲ protein specific antibody titers by SDS-PAGE, Western blotting, indirect ELISA and IFA.SDS-PAGE results showed the expressed target protein existed mainly in the form of inclusion body.Western blotting, ELISA test results showed that the protein had good reactivity with anti-serum.The mice immunized with the purified JEV E Ⅲ protein generated 1×10⁵ anti-JEV E Ⅲ protein specific antibody titers by ELISA, and the porcine immunized with the porcine JEV generated 5.1×10⁴ anti-JEV specific antibody titers.The IFA results showed that JEV E Ⅲ protein anti-serum could identify JEV antigen.The above results showed that the recombinant JEV E Ⅲ had good antigenicity.These results provided important basis for development of diagnostic antigen for JEV.     

Isolation and Identification of BPIV3 Genotype C

To identify the infection agents from Ningxia Hui Autonomous region, where feedlot cattle indicated bovine respiratory disease complex (BRDC), the M gene of the bovine parainfluenza virus type 3 was amplified by RT-PCR.The PCR product was ligated to pMD18-T vector and cloned to E.coli DH5α.The positive clones were sequenced and compared with the reference strains in GenBank by the molecular biology software.Sequence alignment results showed that a BPIV3 strain was isolated from the samples and named NX49, the M gene of NX49 included 1 056 nucleotides.Evolutionary analysis showed that the NX49 belonged to BPIV3 C genotype and shared 99.4% nucleotide identity with that of the SD0835 isolated in Shandong province.The characterization of the NX49 demonstrated that it was sensitive to temperature, acid and organic matter.The presence of Mg²⁺ showed no protection against the treatment at high temperature.The HA test suggested that the NX49 enables to agglutinate the guinea pig RBC at 4 ℃ and the titer was 1∶4.The study isolated a BPIV3 genotype C strain successfully, which facilitate the study of molecular evolution and epidemiology of BPIV3 in China.     

Acute and Sub-chronic Toxicity Test on Rats of a New Kind of Antidiarrheal Chinese Herbal Medicine Compound Preparation for Livestock

In order to understand the security of a new kind of antidiarrheal Chinese herbal medicine compound preparation for livestock,acute and sub-chronic toxicity test were conducted.Acute toxicity test used the largest drug dose method,20 Wistar rats were orally treated with the Chinese medicine compound preparation.In the sub-chronic toxicity test,80 rats were randomly divided into 4 groups with 20 rats in each group and orally given a dose of 3 000,1 500,750 and 0 mg/(kg·BW）of Chinese medicine compound preparation once a day for 30 days.The general clinical status was observed,rats weight were measured and the dose was adjusted every week during the test,after the test measured blood routine index,biochemistry index,and preceded the gross anatomy observation,weighing each major organs and calculated the viscera coefficient,and proceded main viscera histopathological observation between the high dose group and the control group.The acute toxicity results showed that every rat would be alive gavaged with the lethal dose（LD₅₀）of compound preparation larger 5 g/(kg·BW).The sub-chronic toxicity autopsy showed that except heart,lung,and testicles in individual rats appeared mild bleeding in the high dose group,the other dose group organs found no abnormal change.The haematological index showed except mononuclear cell rate（P<0.05),and hematocrit declined significantly（P<0.05）in the high dose group,all the indexes of the other groups were in the normal range,there was no significant difference from the control group.The test suggested the Chinese medicine compound preparation was no toxicity under the condition of this test according to acute toxicity classification standard of exogenous chemicals by WTO,there was no effect on the growth and development of rats in the sub-chronic toxicity test,and there was no chronic toxicity at least 1 500 mg/kg feeding conditions in short-term repeated application.     

水氮调控对苜蓿与无芒雀麦混播草地生长和辐射利用的影响

针对我国内陆干旱区人工草地生产管理粗放及气候资源利用不充分等问题,探究合理的牧草种植与水氮供应模式,以充分挖掘区域牧草的生产潜力。采用3 a生(2018年播种)紫花苜蓿(简称‘苜蓿’)和无芒雀麦,分析种植模式(苜蓿与无芒雀麦混播,D1;无芒雀麦单播,D2)、施氮量(低氮量N1:60 kg·hm^-2;高氮量N2:120 kg·hm^-2)和灌水量(以灌水下限占田间持水量θ_f的百分比计,分枝期均充分灌水(75%～85%θ_f),现蕾和初花期轻度亏水W1:65%～75%θ_f、中度亏水W2:55%～65%θ_f、重度亏水W3:45%～55%θ_f,灌水上限均为85%θ_f)对牧草叶面积指数(LAI)、干物质累积量、累积截获光合有效辐射量(CIPAR)、辐射利用效率(RUE)、产量(Y)、耗水量(ET_a)、水分利用效率(WUE)和氮肥偏生产力(PFP_N)的影响。结果表明：(1)牧草LAI和...