猪白细胞介素-2重组蛋白的原核表达及其生物学活性分析

用已构建好的重组表达载体pET30-IL2转化BL-21E.coli,挑取单克隆,利用诱导剂IPTG诱导表达;通过改变诱导剂的浓度、诱导的时间来摸索最佳诱导条件,在最佳诱导条件下大量表达,并利用镍层析柱纯化重组蛋白,用MTS法测定其生物学活性。结果表明,表达产物经SDS-PAGE分析,表达出25ku的融合蛋白,表达产物主要以包涵体的形式存在,最佳的诱导剂浓度为0.1mM,最佳的诱导时间为4h;对表达产物的包涵体处理后,用镍琼脂糖凝胶FF纯化,所得蛋白的纯度约在90%以上。通过透析除去部分尿素,复性后在体外利用MTS法检测其生物学特性,结果表明其仍具有较好的生物学活性,这为进一步大量制备和研究猪IL-2重组蛋白奠定了基础。     

Effects of autophagy inhibitor bafilomycin A1 on macrophages polarization

AIM:To investigate the effect of bafilomycin A1 (Baf A1) on polarization in mouse macrophages RAW264.7. METHODS:The macrophages RAW264.7 were treated with Baf A1, the concentration of M1/M2 polarization related cytokines were determined by ELISA. The markers of M1/M2 polarization in the macrophages were determined by flow cytometry. The formation of autophagicbody was observed by immunofluorescence. Western blot was used to detect the expression of autophagy related protein levels. RESULTS:The concentration of M1 related proinflammatory cytokine tumor necrosis factor-α (TNF-α) was increased significantly after Baf A1 intervention (P<0.01). However, the concentration of M2 related anti-inflammatory cytokines IL-10 and IL-13 showed no significant difference. The double positive rate of CD197 and HLA-DR (M1 marker) positive cells in Baf A1 treated group were significant higher than that in control group (P<0.05), indicated that Baf A1 induced polarization of macrophage to M1. The results of immunofluoresence showed that the autophagosomes formation was notable increased in Baf A1 group (P<0.05), meanwhile Western blot results showed the expression of autophay related protein LC3-Ⅱ but not P62 was marked up-regulated (P<0.05). For autophagy activator rapamycin (Rapa) treated group, autophagosome formation was also significant increased (P<0.05), but the expression of P62 was notable down-regulated (P<0.05). CONCLUSION:Baf A1 induces polarization of mouse macrophage RAW264.7 to M1, which may be related to the inhibitory effect on the formation of autolysosome.     

感应热处理工艺对拖拉机轴类零件扭转强度的影响

对驱动轮工艺轴进行了7种工艺方案的感应热处理表面强化,然后对工艺轴进行扭转破坏试验。通过对驱动轮工艺轴扭转断裂的研究,为驱动轮轴及承受扭力的轴类零件结构设计、工艺流程设计和热处理技术要求提供指导,从而提高其扭转疲劳强度和使用寿命。     

Evaluation on Tolerance of Montmorillonite as a Mycotoxin Adsorbent

In order to study the tolerance dose of Kunming mice to montmorillonite as a mycotoxin adsorbent in diets,the experiment selected 60 healthy male Kunming mice with initial average body weight of (17±0.12)g,randomly divided them into 5 treatments with two replicates per group and 6 mice per replicate.Five experimental diets were added to 0 (control group),0.5%,1%,2% and 5% montmorillonite,respectively.After 30 days,the growth performance,nutrients apparent digestibility,serum biochemical indices and internal organization were tested to determine the tolerance dose of montmorillonite for Kunming mice.The results showed that the ADFI was significantly increased in 0.5% group (P< 0.05),while there were no significant differences in ADFI and ADG between 1% group and control group (P> 0.05),then the ADFI was significantly decreased with the increasing of montmorillonite supplemental level (P< 0.05).The ADG in 2% group had no significant difference when compared with the control group (P> 0.05),while significantly lower than that in 0.5% and 1% groups (P< 0.05).The apparent digestibility of DM,CP and EE showed no significant effect at any level of montmorillonite (P> 0.05).Serum ALT,AST activities and MDA content were gradually increased with the montmorillonite supplemental level increasing,and that in 1%,2% and 5% groups were significantly higher than that in control group (P< 0.05),but there were no significant differences between 0.5% group and control group (P> 0.05).Tissue sections showed that mice livers were damaged in varying degrees when the montmorillonite supplemental level was 5%.In conclusion,based on a comprehensive consideration of all indexes,it could be seen that the tolerance dose of the tested montmorillonite for Kunming mice was between 0.5% and 2% when mycotoxin contents in diets were in security range.     

CO_2加富对不同砧木嫁接黄瓜农艺性状及净光合速率的影响

为明确北方高寒地区普通日光温室内增施CO_2对温室嫁接黄瓜生长及光合特性的影响,以白籽南瓜和黑籽南瓜嫁接的黄瓜为材料,设CO_2(600±50)μL/L(B1)、(800±50)μL/L(B2)、(1 000±50)μL/L(B3)、(350±50)μL/L(CK)4个浓度。试验结果表明,同一生育期内,随着CO_2浓度增加,白籽南瓜和黑籽南瓜嫁接的黄瓜株高、茎粗、叶面积、叶片叶绿素SPAD值和净光合速率均呈先增加后降低趋势,总体表现为B2处理B3处理B1处理CK,即(800±50)μL/L为最适宜CO_2施用量;CO_2加富可以促进白籽南瓜和黑籽南瓜嫁接黄瓜苗生长,但2种嫁接黄瓜对CO_2敏感性不同,黑籽南瓜嫁接的黄瓜形态指标、叶片叶绿素SPAD值和净光合速率明显优于白籽南瓜嫁接的。     

农村户用沼气服务体系评价建模及应用——基于2010年壳牌中国能源调查河南省偃师市调研项目

作为新能源的沼气具有巨大的应用前景.近年来,在我国农村沼气事业的发展过程中出现了以服务体系不完善为主的诸多问题,如何科学、合理地评价农村沼气服务体系水平,发现原因及时解决,对我国农村能源战略的发展具有重要的现实意义.本文首先根据实地调研情况并使用专家咨询法构建了服务体系评价的指标体系；其次,建立了基于三角白化权函数的灰色评价模型；最后根据调研数据,运用评价模型分析了偃师市农村沼气服务体系的现状.     

离子阱液质联用仪应用于植物叶片游离氨基酸快速分析方法的建立

建立一种液相色谱-离子阱串联质谱(LC-MS-MS)同时分析植物组织中23种游离氨基酸的方法。植物样品经盐酸-乙醇法一步提取后直接进样,无需进行衍生和固相萃取等其他前处理步骤。液相色谱采用Intertsil OSD-4C18(250mm×3.0mm)色谱柱,采用甲醇-甲酸体系进行洗脱,对23种氨基酸获得比较理想的分离效果。结果表明,该方法通用性良好、样品制备简单(一步提取,无需衍生)、灵敏度高、检出范围宽、高效快速,可以在30min内完成整个分析过程。     

贵州省黔东南地区草地贪夜蛾寄生性天敌资源调查初报

草地贪夜蛾Spodoptera frugiperda(J. E. Smith)于2019年5月入侵贵州黔东南地区,严重威胁当地农业生态。黔东南地区丰富的生物资源是挖掘草地贪夜蛾自然天敌的重要基础。为明确黔东南地区草地贪夜蛾天敌的发生情况,2019年5月至7月,采用五点取样法对选取的3个地点开展调查。结果表明,草地贪夜蛾在黔东南地区的主要寄生性天敌有5类,分别为白僵菌属、绿僵菌属、茧蜂科、姬蜂科和黑卵蜂属昆虫,寄生率最高分别为45.50%、57.00%、3.00%、20.00%和12.50%,寄生草地贪夜蛾幼虫期4种天敌综合控制力最高达96.50%。本研究可为草地贪夜蛾天敌资源挖掘与利用提供重要参考。     

用模糊综合评判法预测马尾松毛虫幼虫高峰期发生量

为了提高马尾松毛虫一代、二代幼虫高峰期发生量预报的准确性,为有效防治马尾松毛虫提供科学依据,本文运用模糊综合评判的6个数学模型预测安徽省潜山县马尾松毛虫一代、二代幼虫高峰期的发生量,验证预报1989年、1994年、2002年和2017年一代马尾松毛虫幼虫高峰期发生量,预报结果分别为2级、4级、2级和1级,与实况级别完全吻合,预报结果准确。预报1989年、1994年、2002年和2017年二代马尾松毛虫幼虫高峰期发生量,预报结果依次是4级、5级、2级和1级。同样与实况级别全部相同。预报的准确率为100%,模糊综合评判法是一个运算简便、准确性高的预报方法。