鸭MyoG基因编码区的克隆及其在鸭心肌组织发育中的表达研究

肌细胞生成素(myogenin,MyoG)在胚胎发育中发挥作用,并能与在心脏发育过程中起重要作用的MEF2基因家族协同促进肌肉的分化,而其在心肌发育中的作用却鲜有报道。为研究其在胚胎心肌组织发育过程中的作用,本研究扩增了鸭MyoG基因CDS序列,并采用实时荧光定量PCR法检测了MyoG在鸭E10、E14、E18、E22、E27 d及出生后1周雏鸭(P7 d)心肌组织中共6个阶段的表达变化。基因扩增得到鸭MyoG基因CDS序列共684bp,编码227个氨基酸,预测其含有碱性螺旋-环-螺旋(bHLH)结构域。结果表明:MyoG在鸭E10 d中的表达量最高,显著高于E14、E18、E22和E27 d(P0.05);E14 d与E18 d,E22 d和E27 d之间的表达差异不显著(P0.05);MyoG在鸭出壳前心肌组织中的表达量整体呈现下降趋势,出生后1周,表达量又明显上升,与E22 d和E27 d差异显著(P0.05)。MyoG在鸭心肌组织的发育中起作用,其机理可能是通过MyoG的bHLH结构域直接或以MEF2基因家族介导间接实现对心脏发育的调控。     

抗原表位研究方法进展

抗原表位是抗原分子中的主要功能单位,能有效刺激机体的细胞免疫和体液免疫。随着免疫学和生物信息学技术的不断发展,用于研究T细胞表位和B细胞表位的方法得到了很大的提高。论文中概述了近几年用于研究T细胞表位的预测方法和鉴定方法,以及在B细胞表位研究中所用的表位肽扫描技术、蛋白质切割法、噬菌体展示技术、X-衍射与核磁共振、表位预测方法等技术,并对每种研究方法进行了比较,为从事抗原表位研究的人员提供参考,从而更有利于表位肽疫苗的研制和诊断方法的建立。     

洛阳生生牧业犊牛健康管理经验

近年来,牧场逐步走向现代化、规模化发展,生产水平大幅度提高,后备牛的培育也提升到了新的高度。犊牛健康管理是牧场成败的关键,但如何做好犊牛健康管理,成为制约奶牛业健康发展的瓶颈之一。为此,本文对河南洛阳生生牧业有限公司的犊牛健康管理工作进行了认真总结,旨在了解犊牛管理现状,分析存在的问题,探索适合大型牧场应用的犊牛管理模式,促进奶业健康、可持续发展。     

柳树叶饲料化调制加工技术研究

为了探索垂柳(Salix babylonica)树叶的饲料化加工技术,在研究6个生育期(展叶期、开花初期、飞絮期、青绿期、枯黄期、落叶期)柳树叶营养物质含量变化的基础上,分别将其调制成青贮饲料,测定并比较不同生育期柳树叶制备青贮饲料的营养物质含量及发酵品质指标,并对枯黄期和落叶期树叶进行添加乳酸菌菌剂青贮对比试验。结果表明,根据原料营养物质判断,展叶期柳树叶营养品质与优质豆科牧草品质近似,但是随着生育期的延迟,柳树叶营养物质含量和饲用价值显著下降。将不同生育期柳树叶直接青贮之后,无品质改善效果,青贮饲料发酵品质差,饲用价值不大。但是,添加乳酸菌菌剂后调制的柳树叶青贮饲料发酵品质得到了显著改善。综上表明,柳树叶如果直接饲喂利用,最好选择在较为幼嫩的展叶期;如果开发利用其他生育期或枯落后的柳树叶,以添加乳酸菌菌剂调制成青贮饲料为好。     

紧实度对青贮玉米有氧稳定期发酵品质、微生物数量的效应研究

本研究旨在分析不同紧实度对全株玉米青贮有氧暴露期间发酵品质和微生物变化的影响,为生产实践选择适宜的青贮紧实度提供参考。以新饲玉10号青贮玉米为材料,发酵装料密度设计为5个梯度(350,400,500,600,700 kg/m³),发酵期为50 d,检测开窖后各紧实度处理第12,24,36,60,108 h青贮发酵品质和主要微生物的变化,并用多通道温度记录仪监测温度变化。结果表明,有氧暴露时间与紧实度的交互作用对pH、乳酸、乙酸和氨态氮含量以及乳酸菌、霉菌、酵母菌和好氧细菌数量产生极显著影响(P<0.01)。开窖108 h后,紧实度为600 kg/m³青贮饲料的乳酸菌数量最多,达到8.17 lg cfu/g FW,其pH值和氨态氮含量最低,霉菌和酵母菌数量最少,分别为5.38和7.72 lg cfu/g FW;且紧实度600 kg/m³有氧暴露后稳定的时间显著高于其他处理(P<0.05),达到100 h。通过综合评价,在有氧暴露后紧实度为600 kg/m³的全株玉米青贮发酵品质及有氧稳定性最好,建议600 kg/m³为全株玉米最佳青贮紧实度。     

多头带绦虫Tm16与Tm18抗原基因的克隆及原核表达

为原核表达Tml6和Tm18重组蛋白,本研究以自然感染羊源脑多头坳原头节基因组DNA为模板分别扩增Tml6和Tm 18杭原基因全基因片段,测序鉴定后合成其开放阅读框架DNA片段,将基因组序列中的内含子去除并对其稀有密码子进行改造优化,构建重组表达质粒pGEX-Tm 16和pGEX-Tml8.转化大肠杆菌BL21后以IPTG诱导表达,SDS-PAGE分析表达产物并进行纯化.结果显示:在大肠杆菌中表达出带有GST标签的大小约为39.6 ku和39.4 ku的重组蛋白,经谷胱甘肤琼脂糖树脂纯化得到高纯度的可溶性的GST-Tm 16及GST-Tml8重组蛋白,western blot分析表明重组蛋白均能够被兔抗GST单克隆杭体特异性识别.     

13种蒲公英属植物核型似近系数聚类分析

为探究蒲公英属植物亲缘关系,本研究对13种蒲公英属（Taraxacum. F. H. Wigg.）植物进行了核型似近系数聚类分析,并计算了物种间遗传距离。结果表明：当核型似近系数接近0.6960时,13种蒲公英可以聚为4大类群;当核型似近系数接近0.4641时,13种蒲公英共属一类。异苞蒲公英与狭戟片蒲公英的亲缘关系最近,其似近系数最大,为0.9969,核型进化距离最小,为0.0031;多裂蒲公英和亚洲蒲公英的亲缘关系最远,似近系数为0.3343,核型进化距离为1.0959。本研究结果为蒲公英属植物的亲缘关系辨析、种质鉴定及物种进化分析提供依据。     

Effects of Mixed Caulis Lonicerae japonicae Extracts on Fracture Healing and Anti-inflammation

This experiment was aimed to study the effects of mixed Caulis Lonicerae japonicae extracts on fracture healing and inflammation. Rabbit right radius fracture model was induced by dental drill in order to study the medicine's effect on fracture healing. Moreover,the contents of Ca,P and alkaline phosphatase in serum were measured to explore the influences of mixed Caulis Lonicerae japonicae extracts on serum biochemical indexes of fractured rabbits.RAW264.7 cell was induced by LPS in order to research the effect of medicine on inflammation. The concentrations of TNF-α,IL-1β,IL-6 and NO were measured by ELISA. The results showed that the rabbit fracture model was successfully prepared by dental drill, and the extracts of Caulis Lonicerae japonicae could increase the levels of Ca and alkaline phosphatase and reduce the P content in the serum of fractured rabbits. The TNF-α,IL-1β and NO levels in RAW264.7 cell induced by LPS were significantly inhibited by the extracts with concentrations ranging from 50 to 200 μg/mL (P<0.05). However, the extracts had no significant effect on the level of IL-6 (P>0.05). These results demonstrated that the extracts of mixed Caulis Lonicerae japonicae could promote the fracture healing and possessed good resistance inflammation effect.     

Ethyl acetate fraction of flavonoids from Polygonum hydropiper L. modulates pseudorabies virus-induced inflammation in RAW264.7 cells via the nuclear factor-kappa B and mitogen-activated protein kinase pathways

Pseudorabies virus (PRV) infection leads to severe inflammatory responses and tissue damage, and many natural herbs exhibit protective effects against viral infection by modulating the inflammatory response. An ethyl acetate fraction of flavonoids from Polygonum hydropiper L. (FEA) was prepared through ethanol extraction and ethyl acetate fractional extraction. An inflammatory model was established in RAW264.7 cells with PRV infection to evaluate the anti-inflammatory activity of FEA by measuring cell viability, nitric oxide (NO) production, reactive oxygen species (ROS) release, and mRNA expression of inflammatory factors, inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2). Its functional mechanism was investigated by analyzing the phosphorylation and nuclear translocation of key proteins in the nuclear factor-kappa B (NF-κB) and mitogen-activated protein kinase (MAPK) signaling pathways. Our findings indicate that PRV induced inflammatory responses in RAW264.7 cells, and the responses were similar to that in lipopolysaccharide (LPS)-stimulated cells. FEA significantly suppressed NO synthesis and down-regulated both expression and secretion of COX-2, iNOS, and inflammatory cytokines (P<0.05 or P<0.01). FEA also reduced NF-κB p65 translocation into the nucleus and decreased MAPK phosphorylation, indicating that the NF-κB/MAPK signaling pathway may be closely related to the inflammatory response during viral infection. The findings suggested the potential pharmaceutical application of FEA as a natural product that can treat viral infections due to its ability to mitigate inflammatory responses.     

成对或不成对反转试验设计

本文系统探讨了反转试验（交叉设计）设计的数学模型。统计分析方法和原理以及设计要求，根据t-测验思想，给出了成对或不成对反转试验设计的原理和方法。