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.     

玉米霜霉病的侵染途径及防治措施

系统综述了玉米霜霉病的症状表现、发生规律和防治措施等方面的研究进展.加强进境植物检疫、及时清除病残体和选育高抗的玉米品种是防治玉米霜霉病的重中之重.     

绵羊卵泡发育过程中关键信号通路研究进展

卵泡从原始卵泡发育为成熟卵泡，直至排卵、黄体发育等过程都受到精密的调控，产生大量的优势卵泡是绵羊产多羔及实现快速扩繁的关键因素。研究发现，相关信号通路和转录因子通过影响绵羊卵泡中卵母细胞、颗粒细胞的生长，进而调控卵泡的发育成熟，对这些信号通路进行深入了解，有助于探索卵泡发育的调控机制，早日实现绵羊高效繁育。Notch是卵泡发育过程中发挥重要作用的高度保守信号通路，PI3K/AKT/mTOR信号通路各成员都是广泛存在于细胞内的信号转导分子，在卵泡发育早期发挥了主要作用，还有间隙连接（gap junction，GJ）和跨带突触（transzonal projections，TZPs）等物理连接方式，在细胞间的交流通讯起到重要作用。作者详细介绍了Notch信号通路、PI3K/AKT/mTOR信号通路、间隙连接及跨带突触的结构功能在绵羊卵泡发育中的调控作用，为进一步探明绵羊卵泡发育的调控机制提供参考。     

Cotton Research Center,Shandong Academy of Agricultural Sciences,Jinan 250100, China

During the long process of evolution, plants have adapted to their environments through the biosynthesis of secondary metabolites (SM) using the acetate-malonate pathway, mevalonic acids pathway, or shikimic acid pathway, leading to resistance to different pests. As a major cash crop, cotton can produce many secondary metabolites with insecticidal activity, and show self-defense mechanisms under biotic stress conditions. These characteristics of cotton ensure reduced pest incidence and maintenance of ecological balance. This paper discusses the types of SMs, their synthesis, and insect resistance of cotton secondary metabolites in relation to their defense function. We propose possible pest-control strategies using secondary metabolites in cotton.     

番鸭产蛋各期肝脂肪酸组成及AMPK信号通路基因表达研究

旨在探究产蛋各期番鸭肝腺苷酸活化蛋白激酶（AMPK）信号通路基因表达和肝脂肪酸组成,为番鸭肝脂质代谢应答产蛋提供机理研究。本研究选取开产前22周龄、产蛋初期30周龄、产蛋中期40周龄和产蛋末期60周龄母番鸭各15羽,全自动生化仪测定血脂水平,苏木精-伊红（HE）染色和油红O染色观察肝组织学结构,实时荧光定量PCR （qRT-PCR）检测肝AMPK通路基因表达,气相色谱-质谱联用法（GC-MS）检测产蛋各期肝脂肪酸组成。结果表明,总胆固醇（TC）、低密度脂蛋白胆固醇（LDL-C）、甘油三酯（TG）和极低密度脂蛋白胆固醇（VLDL-C）水平在40周龄显著高于22、30和60周龄（P<0.05）;高密度脂蛋白胆固醇（HDL-C）水平在30和40周龄显著低于22和60周龄（P<0.05）。肝HE和油红O染色切片显示,肝在22周龄呈实质状,至产蛋40和60周龄,肝脂滴沉积明显（P<0.05）。AMPKα1在22、30、40和60周龄呈本底低水平表达,并显著低于产蛋各期肉碱脂酰转移酶1（CPT1）和脂肪酸合成酶（FAS）的表达量（P<0.05）,FAS在22、40和60周龄均呈高水平表达（P<0.05）;羟甲基戊二酰辅酶A还原酶（HMGR）、肝细胞核因子4α（HNF4α）、乙酰辅酶A羧化酶1（ACC1）和固醇调控元件结合蛋白-1（SREBP1c）在40周龄表达量显著高于22和30周龄（P<0.05）。肝中饱和脂肪酸（SFA）、单不饱和脂肪酸（MUFA）和多不饱和脂肪酸（PUFA）主要由C16∶0、C18∶0、C18∶1和C20∶2 n-6构成,分别占总脂肪酸含量的32%、16%、30%和9%。C14∶0和C16∶0含量在40周龄显著高于22周龄（P<0.05）;C24∶0、C20∶2 n-6和PUFA含量在60周龄显著高于22和40周龄（P<0.05）。综上,番鸭产蛋期肝通过上调FAS等脂质合成基因表达,合成大量长链脂肪酸,沉积于肝,并增加血脂水平。     

Fas对镉激活PC12细胞线粒体凋亡通路的影响

旨在探究死亡受体Fas在镉致大鼠肾上腺嗜铬细胞瘤细胞（PC12）凋亡中的作用及其对线粒体通路的调控机制,用10 μmol·L^-1镉处理Fas基因沉默的PC12细胞株12 h,通过Western blot检测BH3相互作用域死亡激动剂（BID）、半胱氨酸蛋白酶-9（caspase-9）、半胱氨酸蛋白酶-3（caspase-3）、多聚二磷酸腺苷核糖聚合酶（PARP）的活化情况,Bcl-2相关X蛋白（Bax）、B细胞淋巴瘤/白血病-2基因（Bcl-2）、凋亡诱导因子（AIF）、核酸内切酶G（Endo G）的表达情况,以及细胞色素C（Cyt C）在细胞内的分布情况,免疫荧光染色检测AIF核转位。结果显示,镉极显著上调tBID/BID比值和Bax/Bcl-2比值,诱导Cyt C从线粒体释放到细胞质,激活caspase-9、caspase-3和PARP,增加AIF和Endo G蛋白表达水平（P<0.01）,并诱导AIF核转位;沉默Fas极显著抑制镉引起的tBID/BID比值和Bax/Bcl-2比值升高,Cyt C从线粒体释放到胞浆,caspase-3、PARP蛋白活化和AIF、Endo G蛋白表达水平极显著升高（P<0.01）,显著抑制镉激活的caspase-9（P<0.05）,并抑制AIF核转位。综上表明,Fas通过调控线粒体通路参与镉致PC12细胞凋亡。     

MSTN基因对牛肌动蛋白细胞骨架调节通路的影响

为探究肌肉生长抑制素（myostatin,MSTN）对牛骨骼肌生长发育的作用机制,本研究前期利用定量蛋白质组学与磷酸化蛋白质组学分析野生型蒙古牛（MG.WT）和MSTN^+/-蒙古牛（MG.MSTN^+/-）腿臀肌肌肉组织中蛋白质水平和磷酸化修饰水平的差异变化,使用已建立的牛骨骼肌卫星细胞体外诱导成肌分化模型,检测设计合成的MSTN siRNA （si-MSTN）干扰效果;采用实时荧光定量PCR和Western blotting方法检测转染si-MSTN的增殖期（GM）和分化第3天（DM3）牛骨骼肌卫星细胞中肌动蛋白细胞骨架调节通路相关基因的mRNA和蛋白水平的表达变化,研究敲低MSTN表达对肌动蛋白细胞骨架调节通路的影响。结果显示,在MSTN^+/-蒙古牛肌肉组织中共鉴定到16个肌动蛋白细胞骨架调节通路相关基因表达丰度上调;转染si-MSTN细胞中的MSTN表达水平极显著降低（P<0.01）;在转染si-MSTN的GM期牛骨骼肌卫星细胞中,肌动蛋白细胞骨架调节通路相关基因ENAH、ACTN4和Cdc42的mRNA水平均显著升高（P<0.05）,PFN1、RhoA和ACTN4的蛋白水平均显著或极显著升高（P<0.05;P<0.01）;在转染si-MSTN的DM3牛骨骼肌卫星细胞中,ENAH、CFL1、SCIN和Cdc42基因mRNA水平均显著升高（P<0.05）,RhoA基因mRNA水平极显著升高（P<0.01）,PFN1和ACTN4的蛋白水平均显著升高（P<0.05）。结果表明,干扰MSTN可以促进肌动蛋白细胞骨架调节通路相关基因的表达,探明了MSTN可能通过介导肌动蛋白细胞骨架调节通路影响牛骨骼肌卫星细胞增殖和成肌分化的分子机制,为进一步研究MSTN对牛成肌分化的调控机制提供参考。     

外源ACC通过激活乙烯合成及信号转导诱导小豆抗锈性

大量研究表明, 乙烯可激发植物对死体营养型真菌的抗性, 但我们前期研究发现, 乙烯合成前体ACC可提高小豆对活体营养型真菌——锈菌的抗性, 为初步明确其机制, 本研究分析了ACC处理对小豆乙烯合成及信号转导的影响, 结果表明, ACC处理显著提高了乙烯合成基因VaACS1及信号通路关键基因VaEIN2?VaEIN3?VaERF5的表达水平?此外, ACC处理后再接种锈菌, 小豆锈病的发病程度显著降低?对接种锈菌后不同时间VaPR2和VaPR4的表达分析表明, 相比ACC处理后不接种对照, ACC处理后再接种锈菌的处理, 接种后1~5 d这两个基因表达量显著升高; 与水处理不接种锈菌相比, 水处理接种锈菌5~8 d后VaPR2和VaPR4的表达量虽显著上调, 但应答时间较ACC处理滞后, 且总体表达水平低, 表明ACC激活乙烯通路进而诱导防卫反应基因上调表达是其诱导小豆抗锈性的关键?     

Mechanical Signalling in Plant: Perception and Transduction Pathway

The metabolism and development of plants are controlled through interplay between genetic and environmental cues.For optimal growth plants require the ability to coordinate complex developmental processes and at the same time sense and respond to environmental stimuli.The responses can be divided into three main events: stimulus perception,generation and transmission of a signal,and subsequent changes in downstream biochemical processes.Because of space limitation this review does not provide an exhaustive account of plant signal transduction.Rather,the authors intention is to cover new and important results in this field,including the mechanochemical transform mediated by integrin and integrin-like protein,the change of membrane fluidity respond to stimulus,and the wide accepted transduction pathway of calcium signaling system.The research emphasis and tendency are prospected.