巨噬细胞激活及钙作用的初步研究

经TG诱发的小鼠腹腔渗出液的巨噬细胞及人的巨噬细胞样细胞株U937受PAF(100ng ml).Zymosan A(0.25mg ml).Can A(50μg ml)LPS(1μg ml)等作用后.能引起胞内游离Ca~(2-)浓度的增加,巨噬细胞内酸性磷酸酶增多,细胞骨架更为舒展、丰满.胞内游离Ca~(2-)的增加是由于胞内钙库的释放与胞外钙的内流.上述困子作用后,可使巨噬细胞产生呼吸爆发.其中.Zymosan A的作用尤为强烈.同时还出现胞膜流动性的降低、当胞外环境中有Ca~(2-)时.可增强巨噬细胞的呼吸爆发.以上提示:在巨噬细胞的激活中Ca~(2-)具有重要的作用.     

Mossy cell synaptic dysfunction causes memory imprecision via miR‐128 inhibition of STIM2 in Alzheimer's disease mouse model

Recently, we have reported that dentate mossy cells (MCs) control memory precision via directly and functionally innervating local somatostatin (SST) inhibitory interneurons. Here, we report a discovery that dysfunction of synaptic transmission between MCs and SST cells causes memory imprecision in a mouse model of early Alzheimer's disease (AD). Single‐cell RNA sequencing reveals that miR‐128 that binds to a 3′UTR of STIM2 and inhibits STIM2 translation is increasingly expressed in MCs from AD mice. Silencing miR‐128 or disrupting miR‐128 binding to STIM2 evokes STIM2 expression, restores synaptic function, and rescues memory imprecision in AD mice. Comparable findings are achieved by directly engineering MCs with the expression of STIM2. This study unveils a key synaptic and molecular mechanism that dictates how memory maintains or losses its details and warrants a promising target for therapeutic intervention of memory decays in the early stage of AD.     

三种淡水鱼类在中国南北两个地区的肠道菌群差异比较

山东省德州市禹城市和湖南省长沙市望城区分别属于我国北方和南方地区,环境和气候差异明显。本试验以禹城市和望城区两地的草鱼(Ctenopharyngodon idella)、鲫鱼(Carassius auratus)和鲤鱼(Cyprinus carpio)为研究对象,对肠道中的菌群进行16s rRNA V4高变区扩增,基于IonS5 TM XL测序平台进行测序,并对相关数据进行分析。结果表明,在门水平上鲫鱼、鲤鱼、草鱼的肠道优势菌主要为变形菌门(Proteobacteria)、梭杆菌门(Fusobacteria)、硬壁菌门(Firmicutes),拟杆菌门(Bacteroidetes),各组中这四种菌门所占比之和均在80%以上;基于Bray-Curtis距离值的秩次进行组间差异显著性检验,山东草鱼组和湖南草鱼组比较中组间差异大于组内差异(R=0.307,P<0.05),且线性判别分析显示湖南草鱼组的特征性菌群为弧菌属,而山东草鱼组的为梭菌纲,推测特征性菌群差异与环境的影响有关;有害菌种的统计分析表明,湖南鲫鱼组和鲤鱼组的嗜水气单胞菌属相对丰度明显高于对应的山东鲫鱼组和鲤鱼组,山东草鱼组和鲤鱼组的弧菌属相对丰度高于对应的湖南草鱼组和鲤鱼组,可能受外界环境的影响,不同地区抵御外来有害细菌的能力也不一样。本研究对两地的淡水鱼类肠道微生物群落结构进行了比较和分析,为进一步的鱼类发育研究奠定基础。     

地涌金莲MlCCD8b基因的克隆与表达分析

该研究以地涌金莲(黄色苞片型YN01和红色苞片型RD05)为材料,采用RACE技术克隆获得地涌金莲CCD8b基因的cDNA全长,进行氨基酸序列比对及系统进化树构建,并采用实时荧光定量PCR技术检测MlCCD8b基因在不同地涌金莲类型和不同组织中的表达模式。结果表明：(1)序列分析显示,MlCCD8b的ORF全长1 671 bp,编码556个氨基酸,存在1个类胡萝卜素加氧酶家族的典型保守结构域RPE65,推测其相对分子量为61 574.26 Da,等电点6.61,亚细胞定位于叶绿体基质中的类囊体上,所编码的MlCCD8b蛋白为亲水性蛋白。(2)同源对比分析及构建系统进化树发现,地涌金莲MlCCD8b蛋白与小果野蕉亚种、凤梨等单子叶植物的CCD8b蛋白遗传关系最近。(3)荧光定量PCR检测结果表明,MlCCD8b在吸芽数量多的黄色苞片型YN01的所有组织中均有表达,而在吸芽数量少的红色苞片型RD05中,其苞片内未能检测到MlCCD8b的表达,但其他组织中皆有表达;MlCCD8b在2种类型地涌金莲中呈现一致的组织表达特异性,即在花序轴中的表达量最高,其次是吸芽芽点、根尖和叶片,在苞片中的表达量最低或不表达。(4)2种类型地涌金莲同一组织部位比较结果显示,RD05的花序轴、吸芽芽点、根尖和叶片中的MlCCD8b相对表达量分别是YN01的4.47、4.67、2.09和1.10倍。(5)利用高效液相色谱 串联质谱法测得RD05根尖部位的5 脱氧独脚金醇含量是YN01的15.57倍,与MlCCD8b在根尖的表达趋势一致。研究认为,MlCCD8b基因可能通过调控独角金内酯的合成,促进或抑制地涌金莲吸芽的萌生。该研究结果可为MlCCD8b基因的生物学功能研究提供依据,并为今后通过分子辅助育种调控MlCCD8b的表达,从而控制地涌金莲吸芽数量提供理论支持。     

Homeotic transformation from stamen to petal in Lilium is associated with MADS-box genes and hormone signal transduction

Plant Growth Regulation - As one of the most popular bulbous plant with high ornamental value, Lilium spp. has been used as cut flower or green plants in urban landscapes and home gardening. We...     

DNA sequencing,genomes and genetic markers of microbes on fruits and vegetables

The development of DNA sequencing technology has provided an effective method for studying foodborne and phytopathogenic microorganisms on fruits and vegetables (F & V). DNA sequencing has successfully proceeded through three generations, including the tens of operating platforms. These advances have significantly promoted microbial whole-genome sequencing (WGS) and DNA polymorphism research. Based on genomic and regional polymorphisms, genetic markers have been widely obtained. These molecular markers are used as targets for PCR or chip analyses to detect microbes at the genetic level. Furthermore, metagenomic analyses conducted by sequencing the hypervariable regions of ribosomal DNA (rDNA) have revealed comprehensive microbial communities in various studies on F & V. This review highlights the basic principles of three generations of DNA sequencing, and summarizes the WGS studies of and available DNA markers for major bacterial foodborne pathogens and phytopathogenic fungi found on F & V. In addition, rDNA sequencing-based bacterial and fungal metagenomics are summarized under three topics. These findings deepen the understanding of DNA sequencing and its application in studies of foodborne and phytopathogenic microbes and shed light on strategies for the monitoring of F & V microbes and quality control.     

Thirty-five percent oxygen pre-conditioning protects PC12 cells against death induced by hypoxia

The present study is designed to investigate the effect of pre-conditioning with 35% O₂ on PC12 cell death induced by hypoxia. This study investigated whether 35% O₂ pre-conditioning for 3 h, followed by 12 h recovery, can protect PC12 cells against death induced by subsequent exposure to hypoxia for 72 h. The result showed that pre-conditioning with 35% O₂ partly blocked the decrease in 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction induced by hypoxia in PC12 cells. PC12 cells pre-conditioned with 35% O₂ could generate a small quantity of reactive oxygen species (ROS), which activated the extracellular signal-regulated kinase (ERK) signalling pathway, then the over-expression of the B-cell lymphoma/leukaemia-2 (Bcl-2) was induced, which subsequently protected PC12 cell against death resulting from hypoxia exposure. In conclusion, 35% O₂ pre-conditioning could protect PC12 cells against hypoxic insult.     

UFD4 lacking the proteasome-binding region catalyses ubiquitination but is impaired in proteolysis

The ubiquitin system recognizes degradation signals of protein substrates through E3-E2 ubiquitin ligases, which produce a substrate-linked multi-ubiquitin chain. Ubiquitinated substrates are degraded by the 26S proteasome, which consists of the 20S protease and two 19S particles. We previously showed that UBR1 and UFD4, two E3 ligases of the yeast Saccharomyces cerevisiae, interact with specific proteasomal subunits. Here we advance this analysis for UFD4 and show that it interacts with RPT4 and RPT6, two subunits of the 19S particle. The 201-residue amino-terminal region of UFD4 is essential for its binding to RPT4 and RPT6. UFD4(DeltaN), which lacks this N-terminal region, adds ubiquitin to test substrates with apparently wild-type activity, but is impaired in conferring short half-lives on these substrates. We propose that interaction of a targeted substrate with the 26S proteasome involves contacts of specific proteasomal subunits with the substrate-bound ubiquitin ligase, with the substrate-linked multi-ubiquitin chain and with the substrate itself. This multiple-site binding may function to slow down dissociation of the substrate from the proteasome and to facilitate the unfolding of substrate through ATP-dependent movements of the chaperone subunits of the 19S particle.     

Identification of tripeptides recognized by the PDZ domain of Dishevelled

The development of inhibitors of Dishevelled (Dvl) PDZ protein–protein interactions attracts attention due to a possible application in drug discovery and development. Using nuclear magnetic resonance (NMR) spectroscopy, we found that a tripeptide VVV binds to the PDZ domain of Dvl, which is a key component involved in Wnt signaling. Using a computational approach calculating the binding free energy of the complexes of the Dvl PDZ domain and each of the tripeptides VXV (X: any amino acid residue except Pro), we found that a tripeptide VWV had the highest binding affinity. Consistent with the computational result, experimental results showed that the binding of the tripeptide VWV to the Dvl PDZ domain was stronger than that of the tripeptide VVV. The binding affinity of the tripeptide VWV was comparable to that of the organic molecule NSC668036, which was the first identified Dvl PDZ inhibitor. The three-dimensional structure of the complex Dvl1 PDZ/VWV was determined to investigate the role of the energetically favorable W(?1) residue in binding. These interactions were also explored by using molecular dynamic simulation and the molecular mechanics Poisson–Boltzmann surface area method. Taken together, these two tripeptides may be used as modulators of Wnt signaling or as a scaffold to optimize an antagonist for targeting Dvl1 PDZ protein–protein interaction.