小鼠H-Y单克隆抗体ELISA检测方法的建立

以纯系 BAL B/ c雄性小鼠脾细胞腹腔注射免疫同系雌性小鼠 9次 ,获得的抗血清经雌、雄鼠脾细胞吸收后用于精子细胞毒性试验 ,测得 H- Y抗血清效价为 1/ 16 0。选取免疫应答最好的雌鼠脾细胞与 SP2 / 0骨髓瘤细胞融合 ,用精子细胞毒性方法筛选效价较高的细胞株制备腹水 ,建立优化的 EL ISA反应条件。优化后的 EL ISA反应条件为 :抗原4℃过夜 ,加 H- Y抗血清 37℃反应 12 0 m in,加 HRP- Ig G 37℃反应 30 min,加 TMB 2 5℃ 30 min。优化后的 EL ISA与常规 EL ISA比较 ,可以明显降低阴性吸光值 ,检测灵敏度从 1/ 32 0上升为 1/ 12 80     

日粮锌、硒水平对肉鸡肠道黏膜屏障结构的影响

为了探讨微量元素锌和硒相互作用对肉鸡肠道黏膜屏障结构的影响，将24只1日龄AA肉鸡随机分3组，分别饲喂添加有高锌高硒(锌1000mg／kg、硒5mg／kg)、低锌低硒(锌34mg／kg、硒0．08mg／kg)或常锌常硒(锌50mg／kg、硒0．15mg／kg)的日粮45d后，观察肠黏膜上皮细胞、上皮内淋巴细胞和盲肠扁桃体的形态结构变化。结果表明：高锌高硒或低锌低硒组肉鸡的肠黏膜结构有明显的损伤，表现为肠黏膜上皮细胞萎缩，绒毛长度下降，上皮内淋巴细胞数量减少；盲肠扁桃体的弥散淋巴组织和淋巴小结中，淋巴细胞数量减少，细胞出现肿胀，有的核消失，结缔组织增生，淋巴小结萎缩。尤其是高锌高硒组的损伤最为严重。而常锌常硒组肉鸡肠黏膜和盲肠扁桃体的形态结构正常。结论：日粮中按锌50mg／kg、硒0．15mg／kg的比例添加，对于维持肠道黏膜的正常屏障结构是合适的。过高或过低的锌和硒对小肠黏膜有毒性作用，破坏其屏障功能；而且高锌和高硒可相互促进以增强其毒性作用。     

Expression and role of CD134 and NF-κB in renal tissue of lupus nephritis

AIM:To investigate the role of CD134 (OX40) and NF-κB in the pathogenesis of lupus nephritis (LN).METHODS:Renal in situ CD134 and NF-κB expression were examined in 40 biopsy specimens from LN patients by immunohistochemistry and microwave-based immunohistochemistry, respectively. The relationship between expression of CD134 and NF-κB was analyzed.RESULTS:The expression of glomerular and tubular CD134 and NF-κB in LN were higher than that in normal control, especially in class Ⅳ LN, where there was intense staining of endothelial cell, distal tubules, and interstitial mononuclear cell. The CD134 expression of glomerular and tubular was closely related to NF-κB expression, respectively (r=0.5542, P＜0.05;r=0.6279, P＜0.05). CONCLUSIONS:The abnormal expression of costimulatory molecule CD134 was well evidenced in LN. Strong expression of renal in situ NF-κB was likely mediated by CD134 signal pathway, which may play an important role in the pathogenesis of LN.     

食用菌复合保水颗粒菌种生产工艺及其应用研究

运用微生物发酵技术，配合新型合成材料，研制出食用菌复合保水颗粒菌种。这类菌种可保持活力60d。这一工艺可用于糙皮侧耳、金针菇、灵芝、白灵菇、杏鲍菇、鸡腿蘑和鲍鱼菇等多种食用菌苗种的制作。栽培试验证明，此类菌种的菌丝生长速度、现蕾时间、生物学效率与常用的棉籽壳菌种相似。     

猪卵泡卵母细胞体外成熟与冷冻保存

研究了激素、猪卵泡液、不同类型血清对猪卵泡卵母细胞体外成熟的影响 ;比较了卵泡直径小于 2 mm、2～ 5 mm和大于 5 mm的卵母细胞体外成熟能力的差异 ,并对不同发育阶段猪卵母细胞的冷冻保存进行了研究。结果表明 :猪卵母细胞体外培养 4 8h时 ,培养的前 2 4 h培养液中加入激素 ,后 2 4 h去掉激素 ,卵母细胞的 A级成熟率 (5 1.73% )和总成熟率 (83.2 5 % )最高 ,极显著高于前 2 4 h不加激素 ,后 2 4 h添加激素培养的成熟率 (P<0 .0 1) ;也显著高于不含激素的培养液连续培养 4 8h的成熟率 (P<0 .0 5 ) ;但与添加激素连续培养 4 8h的成熟率差异不显著 (P>0 .0 5 )。在体外成熟培养液中 ,添加 10 % (体积分数 ) ECS的成熟率 (72 .86 % )显著高于添加 10 % (体积分数 ) NCS的成熟率(6 2 .2 1% ) (P<0 .0 5 ) ,而添加 10 % p FF则抑制卵母细胞的体外成熟。随着卵泡直径的增大 ,卵母细胞体外成熟能力逐渐增强。采用程序冷冻保存方法 ,成熟卵母细胞的成活率 (35 .5 9% )显著高于培养前 (2 4 .6 4 % )和培养 2 4 h(2 3.36 % ) (P<0 .0 5 )。培养 2 4 h(77.2 2 % )和培养成熟 (72 .81% )的卵母细胞解冻后的形态完整率均显著高于培养前(5 3.2 4 % ) (P<0 .0 5 )     

枯草杆菌的淀粉酶基因在大肠杆菌中的表达

根据枯草杆菌(Bacillus subtilis)基因文库中的淀粉酶基因的碱基序列,设计引物。以枯草杆菌菌体DNA为模板,利用PCR扩增出分子量大约为2.3 kb的淀粉酶基因片段。借助于核酸内切酶和连接酶把该基因植入到pHMSXD1质粒中,构建pHMSXD2质粒载体。通过高压电击处理把载体植入到大肠杆菌(JM10)中并成功表达。该株大肠杆菌在酶基因表达后,其淀粉酶活力达到1.037 6 U/mL(LB培养液)和1.361 0 U/mL(矿物元素培养液),分别比原始的枯草杆菌的淀粉酶活力提高了46倍和286倍。     

基于遗传算法的LQG/LTR航空发动机性能优化

针对飞行控制系统控制器的设计中存在建模误差以及飞行过程中外界干扰的影响,采用LQG/LTR鲁棒控制方法完成某型战斗燃油机控制系统控制器的设计。为了提高战斗机控制精度,解决LQG/LTR鲁棒控制方法中存在的权矩阵Q和R选择困难的局限性,加入遗传算法,进行在线寻优。同时针对不同飞行条件下系统以及系统建模过程中造成的参数不确定性问题分别进行仿真，并与传统PID以及控制效果较好的基于遗传算法的PID控制进行对比仿真。理论研究和系统仿真结果表明,基于遗传算法的LQG/LTR控制系统与基于遗传算法的PID控制相比,不但具有良好的鲁棒性,而且响应快速,控制精度高,满足了战斗机飞行控制的要求。     

薄皮甜瓜新品种京玉绿宝的选育

京玉绿宝是以白沙蜜5代自交系357为母本,以地方品种240提纯系选后代397为父本配制而成的薄皮甜瓜一代杂种。果实扁卵圆形,单果质量200~400g,果面光滑无棱,果皮深绿色,果肉白绿色,可溶性固形物含量11%~15%,早熟,抗逆性较强,适合保护地及少雨露地栽培,每667m2产量1600kg。已在北京、河北、辽宁、吉林、内蒙古等地示范推广3000hm2。     

Mycoplasma pneumoniae induces IL-1β production through activating NLRP3 inflammasome by ROS in RAW264.7 cells

AIM: To investigate whether Mycoplasma pneumoniae (Mp)-induced interleukin-1β (IL-1β) production in RAW264.7 cells is through the activation of NLRP3 inflammasome via reactive oxygen species (ROS). ME-THODS: RAW264.7 cells were randomly divided into 3 groups. In normal group, RAW264.7 cells were treated without Mp. In model group, RAW264.7 cells were treated with 1∶ 10 multiplicity of infection (MOI) of Mp. In NAC group, RAW264.7 cells were pretreated with N- acetylcysteine (NAC) at a concentration of 5 mmol/L for 30 min before infection with Mp. The RAW264.7cells were infected with Mp (1∶ 10 MOI) for 4, 8, 16 and 24 h in model group and NAC group, respectively. The intracellular ROS level was analyzed by flow cytometry. The mRNA expressions of NLRP3, ASC and caspase-1 were detected by real-time PCR. The protein levels of NLRP3, ASC and caspase-1 p20 were determined by Western blot. The levels of pro-inflammatory cytokine IL-1β in the supernatant were measured by ELISA. RESULTS: Compared with normal group, the production of ROS were significantly increased at 4, 8, 16 and 24 h after infection, the mRNA expression of NLRP3, ASC and caspase-1 were increased at 8, 16 and 24 h after infection, the protein levels of NLRP3, ASC and caspase-1 p20 were increased at 16 and 24 h after infection, and the releases of IL-1β were increased at 24 h after infection in model group (P<0.01). Compared with the model group, the level of ROS in NAC group decreased, so as the expression of NLRP3, ASC and caspase-1 at mRNA and protein levels and the releases of IL-1β in the supernatant at the corresponding time points. CONCLUSION: Mp may stimulate the ROS production to activate NLRP3 inflammasome in RAW264.7 cells.     

高等院校科研型实验室管理探索——以广东海洋大学食品科学与工程学科为例

探讨高等院校科研型实验室的特点和管理模式,通过健全管理制度,发挥科研项目的引领作用,促进校企合作,建设大型仪器平台,发挥教师、研究生和本科生的作用,提高科研型实验室的管理水平。