Preliminary Study on NF-κB Signaling Pathway Regulating Brucella Intracellular Survival Molecular Mechanisms

By the infection of Brucella virulent strain and attenuated strain in mice macrophage RAW264.7,the assay was aimed to explore the relationship between NF-κB signaling pathways and Brucella virulent strain and attenuated strain in intracellular survival.Use different MOI Brucella (2308,RB51,16M and M5) to infect mice macrophage RAW264.7,after 0,4,8 and 24 h infected,cracking cell and collecting supernatant,we detected the effect of Brucella on activation of NF-κB signaling pathway by Western blotting.Different concentrations of NF-κB signaling pathway inhibitor were incubated with mice macrophage RAW264.7,with different multiplicities of infection (MOI) of Brucella infecting cells,ELISA kits to detect the expressions of TNF-α,IL-1β and IL-6 cytokine;At the same time,count the number of intracellular bacteria of CFU.The results showed that rough cattle Brucella strains RB51 could strongly activate NF-κB signaling pathway,smooth cattle Brucella strains 2308 was weak in the activation;At the same time,the activation of NF-κB signaling pathway was concentration dependent.When the MOI was 80,infection time was 8 h,NF-κB activation degrees of rough cattle Brucella strains RB51 and smooth cattle Brucella strains 2308 were the strongest,and this pathway was involved in producing TNF-α and IL-6;NF-κB signaling pathway inhibitor BAY11-7082 affected Brucella intracellular survival.So rough cattle Brucella strains RB51 intracellular survival and NF-κB signaling pathway activity were closely related.The results laid the foundation for the further study of Brucella intracellular pathogenesis,also provided scientific basis for the research of new drugs to Brucella,and prevention and treatment of brucellosis.     

水稻条纹叶枯病的研究 Ⅳ.病叶细胞的病理变化

在水稻条纹叶枯病叶超薄切片的电镜检查中,可见一些叶肉细胞和维管束鞘细胞中叶绿体有不同程度的降解和淀粉粒累积,一些细胞不同程度坏死,叶肉细胞、维管束鞘细胞以及伴胞的细胞质或液泡中有一个或多个蛋白体存在,一些伴胞中有特异性的非外壳蛋白存在,一些细胞中有一些不定形的粒状内含体或砂状结构,而在病株的不同叶位,这种砂状结构数量不同,以+1叶所见最多.研究结果还佐证了前文有关叶绿体中淀粉粒的过量累积可能引起叶绿体的破坏及其后叶片褪绿病斑形成的假说,并推测这可能是同化产物的运输通道受到影响的结果.     

棉铃虫兼性迁飞的初步研究

本文用吊飞装置测定了棉铃虫成虫不同日龄的飞行能力,测得３日龄雌蛾平均飞行速度为０．９２ｍ／ｓ,平均飞行时间为９．７２±３．６２ｈ,并与其他迁飞性和非迁飞性昆虫作了比较。在实验室条件下研究了温度和食料对生殖前期及繁殖的影响,证实此虫适于迁飞的生理时期较短,最多仅１～２个夜晚。用雌蛾卵巢解剖方法比较了江苏丰县和辽宁朝阳两地二代和三代的虫源性质,分析了两地的发蛾特征。证实朝阳１９９５～１９９６年的二代和部分三代成虫均从外地迁入,但与粘虫不同,迁入种群无突增现象,蛾群稀疏而持续不断。而江苏丰县则在１９９４～１９９６年均无明显迁入现象,以本地虫源为主。讨论了中国棉铃虫的迁飞势能和兼性迁飞的特性。     

STAT6介导的巨噬细胞极化对布鲁氏菌胞内存活的影响

旨在探讨STAT6介导的巨噬细胞极化对布鲁氏菌胞内生存的影响。本研究采用布鲁氏菌光滑株S2308（S2308）和粗糙型疫苗株RB51（RB51）侵染巨噬细胞。利用qRT-PCR检测M1型巨噬细胞标志因子p65、NOS2和IL-1β,M2型巨噬细胞标志因子STAT6、ARG1、IL-10的mRNA表达水平;流式细胞术检测M1型标记分子CD86和M2型标记分子CD206的表达;Western blot检测p-STAT6蛋白及抑制剂AS对蛋白的抑制作用;ELISA检测M1型细胞因子TNF-α、IL-12和M2型细胞因子IL-4、IL-10的表达量;最后对胞内菌落进行CFU计数。qRT-PCR结果显示,在感染8、12 h时可显著诱导M1型因子mRNA转录表达,72 h时低表达,而M2型因子在72 h时高表达;流式细胞术结果显示,S2308感染12 h可显著诱导CD86的表达,感染72 h可显著诱导CD206的表达,但RB51对二者无影响;Western blot结果显示,S2308菌株在感染72 h时激活STAT6信号通路,而RB51几乎不激活该通路,抑制剂AS在2 μmol·L^-1浓度时抑制效果最佳;ELISA结果显示,AS抑制剂可显著抑制IL-4、IL-10的释放,并促进TNF-α、IL-12的释放;CFU计数结果显示,S2308组的胞内菌呈先降低后显著上升趋势,加入AS抑制剂后可显著抑制布鲁氏菌胞内复制。布鲁氏菌S2308在感染后期能够通过STAT6诱导M1型巨噬细胞向M2型转化,并促进Th2型细胞因子的释放,从而有利于布鲁氏菌的胞内生存。而RB51几乎不激活该通路,不影响胞内生存。     

木芙蓉的繁育系统及传粉生物学

以木芙蓉为研究材料,运用杂交指数、花粉数/胚珠数、座果率、种子萌发率等指标对木芙蓉的繁育系统及传粉生物学等进行研究。结果表明:木芙蓉单花的花期1 d,整体花期7—11月份;木芙蓉的杂交指数为4,繁育系统为异交、部分自交亲和、需要传粉者;木芙蓉的花粉数/胚珠数为415±11.3,繁育系统属于兼性异交类型;木芙蓉自然座果率为74%;同株同花授粉的座果率为40%;人工异株异花授粉座果率为71%;各种授粉方式下,单个果实的种子个数为128~148,差别不大;各种授粉方式下,平均种子萌发率为47%~60%,没有显著差异;传粉昆虫有蜂类、甲虫,其中中华蜜蜂、黑小蜜蜂为主要传粉昆虫。     

Acquisition of the potential for sperm motility in steelhead (Oncorhynchus mykiss): effect of pH on dynein ATPase

Salmonid sperm pre-incubated at extracellular pH (pH_e) values less than about 7.4 do not become motile upon water activation whereas sperm maintained above about pH 8.0 demonstrate maximal motility upon activation. The basis for this permissive effect of elevated pH_e on sperm motility is not known. Since it is conceivable that the pH sensitivity of dyneinATPase (the molecular motor that drives flagellar movement) could be the basis of, or contribute to this pH dependency, the pH sensitivity of this enzymatic activity was evaluated in membrane-permeabilized axonemes (isolated flagella) ofsteelhead sperm. DyneinATPase activity was found to be sensitive to pH. This activity in permeabilized axonemes was about 3.5-fold higher at pH 7.6 compared to 7.0. To determine whether the pH sensitivity ofATP regeneration might affect the interpretation of the effect of pH on dyneinATPase activity, the pH sensitivities of creatine kinase and adenylate kinase were established. The rates ofATP generation by these enzymes were insensitive to pH between 6.5 and 8.0. The results of these studies are consistent with the hypothesis that prior maintenance at pH_e, in part, controls the potential for sperm motility upon water activation via an influence on dyneinATPase activity. However, the potential for motility ofsteelhead sperm is particularly sensitive to prior maintenance at pH_e values between about 7.4 and 8.0 whereas the dyneinATPase activity of permeabilized axonemes was particularly sensitive to pH values between 7.0 and 7.6. Phosphorus NMR spectroscopy was used to determine that sperm intracellular pH (pH_i) increased with increasing pH_e between 7.0 and 8.5 and pH_i was, on average 0.4–0.5 pH units lower than pH_e. Therefore the pH_e sensitivity of the potential for motility appears to correspond to the pH_i sensitivity of dyneinATPase activity. The data indicate that pH_i is directly related to pH_e and that prior incubation at pH_e may, in part, control the sperm's potential for motility upon water activation via an influence on dyneinATPase activity.     

Minimising aerobic respiratory demands could form the basis to sub-lethal copper tolerance by rainbow trout gill epithelial cells in vitro

Mechanisms of Cu tolerance were investigated in respiratory epithelial cell cultures, from rainbow trout gills, by studying O₂ consumption and protein synthesis rates, intracellular Na concentration and TER. The lowest concentration found to reduce O₂ consumption was 25 M Cu. This did not affect either protein synthesis rate or intracellular Na concentration and was interpreted in terms of copper tolerance; i.e., how these two energetically demanding processes are maintained despite a reduction in aerobic ATP supply. The relationship between protein synthesis rate and synthesis cost is exponential and the cost of protein synthesis in gill cells was found to be minimal (i.e., this cell occupies a position on the asymptotic section of the protein synthesis rate/synthesis cost model) and unaffected by 25 M Cu. Thus protein synthesis rates could be maintained since any reduction would represent an insignificant energy saving. Intracellular Na concentrations and O₂ consumption rates were linearly correlated suggesting reducing intracellular maintenance costs would have a greater significance in terms of overall energetic conservation. Intracellular Na maintenance costs, calculated from O₂ consumption rates and intracellular Na concentrations, were found to decline after exposure to 25 M Cu. Since TER was unaffected this implied the reduced costs arose from membrane `channel arrest'. Thus the Na/K ATPase energy demands, associated with maintaining intracellular Na concentration, could be reduced by decoupling metabolic demand and membrane function. Therefore this study may demonstrate how the flexibility of cellular energetics enables gill epithelial cells to tolerate sub-lethal Cu.