The Role of Cdk5 in neuronal death and synaptic dysfunctions through phosphorylating FOXO1 and BAG3

细胞周期蛋白依赖性激酶5（Cyclin-dependentkinase5，Cdk5）是由脯氨酸介导的丝氨酸/苏氨酸蛋白激酶，其在神经系统中发挥着重要的作用。Cdk5的活化主要靠神经系统中广泛表达的两个激活因子p35和p39。Cdk5对神经系统的正常生理功能具有重要作用去，其可通过磷酸化众多底物的丝氨酸/苏氨酸位点，在神经元的迁移分化，突触传递，轴突树突的发育过程以及神经元的存活中都发挥着重要的作用。此外，在某些病理条件下，Calpain所介导的p35切割形成p25，可以过度激活Cdk5，诱导神经元凋亡和突触功能的紊乱，介导了多种神经退行性疾病的发生发展，暗示着Cdk5很可能成为治疗神经退行性疾...Cyclin-dependent kinase 5 (Cdk5) is a multifaceted proline-directed serine or threonine kinase implicated to play pivotal roles in nervous system. Cdk5 two activator proteins p35 or p39 ubiquitously expressed in neuronal cells. In the physiological condition, Cdk5 plays vital roles in the CNS development by phosphorylating the specific serine or threonine site of numerous substrate proteins that a...学位：理学博士院系专业：医学院_生理学学号：2452013015433

单晶6H-SiC经氦离子辐照及退火后的微观组织研究

在400℃下对单晶6H-SiC进行了400keV氦离子辐照,辐照剂量为1×1016He+/cm2,随后在1200和1500℃退火30min。采用透射电子显微镜和扫描电子显微镜对辐照态和退火态SiC进行微观结构观察与分析。结果表明,单晶6H-SiC在400℃经氦离子辐照后,仅观察到由辐照引起的位移损伤带,而未观察到明显尺寸的氦气泡。但经1200℃退火30min后,在辐照损伤区域形成了呈血小板状的气泡簇,其主要分布在（0001）晶面上,少量形成在（1120）晶面。辐照未在6H-SiC表面上形成明显尺寸的缺陷,而经1200℃退火30min后,SiC表面出现大尺寸的起泡和凹坑,进一步提高退火温度至1500℃时,表面起泡和形成凹坑更严重,并产生了大量裂纹。本研究同时对微观结构演化的机理进行了分析与讨论。中国工程物理研究院NPL,CAEP项目资助(2015AB001

Expression and test of the neutralization Fab antibody against infectious bursal disease virus

为表达抗鸡传染性法氏囊病病毒(IbdV)抗体fAb并检测其中和活性,本研究将抗IbdV抗体的轻链(l)和重链片段(fd)基因分别克隆于P ET-27b(+)载体中,并转化于大肠杆菌rOSETTA(dE3)进行诱导表达。将l和fd片段包涵体蛋白变性后等量混合于复性液中,制备fAb并对其进行活性鉴定。结果显示l和fd蛋白相对分子质量大小分别为25 ku和28 ku。WESTErn blOT和ElISA检测结果表明,获得的抗体fAb大小约为50 ku,并且与VP2蛋白和不同病毒株均具有特异性结合能力。体外中和试验结果表明,获得的IbdV抗体fAb具有中和活性,可以有效阻断IbdV(b87株)对鸡胚成纤维细胞(df1)的感染。本实验获得的IbdV抗体fAb有望成为治疗Ibd的候选生物制剂,为研制治疗Ibd抗体制剂奠定了基础。To express the neutralizing Fab antibody against infectious bursal disease virus(IBDV),the gene of light chain(L)or heavy chain fragment(Fd) against IBDV was cloned into the prokaryotic expression plasmid,respectively,and then the recombinant L or Fd was expressed in E.coli Rosetta(DE3),respectively,and purified through sole denaturation and co-renaturation of inclusion body.Western blot results showed that the Fab was approximately 50 ku.ELISA results showed that the Fab exhibited binding ability and specify to VP2 for different IBDV strains.The results of neutralization test in vitro showed that the Fab exhibited neutralizing activity to IBDV-B87 strainin chicken embryo fibroblast(DF1) cells.The Fab antibody prepared in this study is expected to become a candidate drug for treatment of IBD,which laid the foundation for the treatment of IBD.黑龙江省应用技术研究与开发计划项目(GC13C104

第十八届美国理论与应用力学大会总结

1会议概况2018年6月5—9日,第18届美国理论与应用力学大会(18th U.S. National Congress of Theoretical and Applied Mechanics, USNCTAM2018)在美国芝加哥召开.本次大会由美国力学国家委员会和中国力学学会联合主办,旨在探讨和交流近四年世界范围内在理论和应用力学领域的基础研究、创新技术的最新进展,吸引了来自世界各地的近千名专家学

CDK5-dependent BAG3 degradation modulates synaptic protein turnover

阿尔茨海默病（AD）是严重威胁人类健康的重大神经系统疾病，AD的发生发展与衰老密切相关，目前临床治疗方法十分有限。因此迫切需要从AD致病早期入手，发现和鉴定导致AD神经功能紊乱的机制和靶点，为AD的早期防治提供基础。张杰教授及其团队从高通量磷酸化蛋白质组学入手，系统研究了CDK5在神经细胞中的磷酸化底物，鉴定出了在蛋白质量控制中发挥重要功能的BAG3蛋白是CDK5的全新底物。课题组从磷酸化蛋白质组学入手，发现和阐明了细胞周期蛋白激酶5（CDK5）通过调控BAG3在维持突触蛋白水平调控中的作用机制，及其在阿尔茨海默病（AD）发生发展中的机理。 该研究是多个团队历时8年合作完成的，香港中文大学的周熙文教授、美国匹兹堡大学的Karl Herrup教授、美国Sanford-Burnham研究所的许华曦教授、美国梅奥医学中心的卜国军教授，厦门大学医学院的文磊教授、张云武教授、赵颖俊教授、薛茂强教授，军事医学科学院的袁增强教授等都参与了该工作。 厦门大学医学院2012级博士生周杰超等为文章的第一作者，张杰教授为通讯作者。Background Synaptic protein dyshomeostasis and functional loss is an early invariant feature of Alzheimer’s disease (AD), yet the unifying etiological pathway remains largely unknown. Knowing that cyclin-dependent kinase 5 (CDK5) plays critical roles in synaptic formation and degeneration, its phosphorylation targets were re-examined in search for candidates with direct global impacts on synaptic protein dynamics, and the associated regulatory network was also analyzed. Methods Quantitative phospho-proteomics and bioinformatics analyses were performed to identify top-ranked candidates. A series of biochemical assays were used to investigate the associated regulatory signaling networks. Histological, electrochemical and behavioral assays were performed in conditional knockout, shRNA-mediated knockdown and AD-related mice models to evaluate its relevance to synaptic homeostasis and functions. Results Among candidates with known implications in synaptic modulations, BCL2-associated athanogene-3 (BAG3) ranked the highest. CDK5-mediated phosphorylation on Ser297/Ser291 (Mouse/Human) destabilized BAG3. Loss of BAG3 unleashed the selective protein degradative function of the HSP70 machinery. In neurons, this resulted in enhanced degradation of a number of glutamatergic synaptic proteins. Conditional neuronal knockout of Bag3 in vivo led to impairment of learning and memory functions. In human AD and related-mouse models, aberrant CDK5-mediated loss of BAG3 yielded similar effects on synaptic homeostasis. Detrimental effects of BAG3 loss on learning and memory functions were confirmed in these mice, and such were reversed by ectopic BAG3 re-expression. Conclusions Our results highlight that neuronal CDK5-BAG3-HSP70 signaling axis plays a critical role in modulating synaptic homeostasis. Dysregulation of the signaling pathway directly contributes to synaptic dysfunction and AD pathogenesis.This work was supported by the National Science Foundation in China (Grant: 31571055, 81522016, 81271421 to J.Z.; 81801337 to L.L; 81774377 and 81373999 to L.W.); Fundamental Research Funds for the Central Universities of China-Xiamen University (Grant: 20720150062, 20720180049 and 20720160075 to J.Z.); Fundamental Research Funds for Fujian Province University Leading Talents (Grant JAT170003 to L.L); Hong Kong Research Grants Council (HKUST12/CRF/13G, GRF660813, GRF16101315, AoE/M-05/12 to K.H.; GRF16103317, GRF16100718 and GRF16100219 to H.-M,C.); Offices of Provost, VPRG and Dean of Science, HKUST (VPRGO12SC02 to K.H.); Chinese University of Hong Kong (CUHK) Improvement on Competitiveness in Hiring New Faculty Funding Scheme (Ref. 133), CUHK Faculty Startup Fund and Alzheimer’s Association Research Fellowship (AARF-17-531566) to H.-M, C. 该研究受到了国家自然科学基金、厦门大学校长基金、福建省卫生教育联合攻关基金等的资助

类泛素蛋白及其中文命名

泛素家族包括泛素及类泛素蛋白,约20种成员蛋白.近年来,泛素家族领域取得了迅猛发展,并已与生物学及医学研究的各个领域相互交叉.泛素家族介导的蛋白质降解和细胞自噬机制的发现分别于2004和2016年获得诺贝尔奖.但是,类泛素蛋白并没有统一规范的中文译名. 2018年4月9日在苏州召开的《泛素家族介导的蛋白质降解和细胞自噬》专著的编委会上,部分作者讨论了类泛素蛋白的中文命名问题,并在随后的\"泛素家族、自噬与疾病\"(Ubiquitinfamily,autophagy anddiseases)苏州会议上提出了类泛素蛋白中文翻译草案,此草案在参加该会议的国内学者及海外华人学者间取得了高度共识.冷泉港亚洲\"泛素家族、自噬与疾病\"苏州会议是由美国冷泉港实验室主办、两年一度、面向全球的英文会议.该会议在海内外华人学者中具有广泛影响,因此,参会华人学者的意见具有一定的代表性.本文介绍了10个类别的类泛素蛋白的中文命名,系统总结了它们的结构特点,并比较了参与各种类泛素化修饰的酶和它们的生物学功能.文章由45名从事该领域研究的专家合作撰写,其中包括中国工程院院士1名,相关学者4名,长江学者3名,国家杰出青年科学基金获得者18名和美国知名高校华人教授4名.他们绝大多数是参加编写即将由科学出版社出版的专著《泛素家族介导的蛋白质降解和细胞自噬》的专家

Emodin Attenuates The Neurotoxicity Induced by Nitric Oxide Through Inhibiting FOXO1 Transcriptional Activity

氧化应激所产生的活性氧和一氧化氮(nitric oxide,NO)自由基在心脑血管疾病、神经退行性疾病中发挥着重要作用,过量的NO可以导致自由基损伤,并诱导神经元的凋亡.大黄作为中医传统药物具有重要的药用价值,临床应用非常广泛.近年研究表明,大黄素具有抗氧化、免疫调节、抗菌、抗炎等功能,被广泛应用于肠道疾病、肾病、心血管疾病、胰腺炎等病症的治疗.Forkhead转录因子1(FOXO1)是Forkhead转录因子家族的一个重要成员,FOXO1对于胰岛素信号通路、DNA修复、清除活性氧损伤、细胞周期和凋亡的调控非常重要.而NO自由基对FOXO1的调控作用还不清楚,我们研究发现,NO的供体GSNO(亚硝基谷胱甘肽)或者L-Arg(L-精氨酸)可显著提高FOXO1的转录活性并促进其下游促凋亡基因Fas L、Bim的转录表达,进而诱导神经元死亡.我们进一步研究发现,大黄素可以通过降低FOXO1的转录水平以及蛋白质水平,缓解NO所诱导的神经元凋亡.该研究揭示了NO自由基诱导神经元损伤的新机制,同时也为了解大黄素的抗氧化作用提供了新的实验依据,对大黄素等中药有效成分的临床应用提供了重要参考.Reactive oxygen species(ROS) and NO free radicals generated from oxidative stress play an important role in the pathogenesis of neurodegenerative disease and cardiovascular and cerebrovascular diseases. Excessive NO production can cause free radical damage and induce neuron death. As one of the traditional Chinese medicine,Emodin have valuable and widely clinical applications. Recently, Emodin has been reported to have antioxidant,immunomodulatory, antibacterial, anti-inflamatory functions etc. FOXO1 is a vital member of the Forkhead family of transcription factors known to regulate the transcription of genes involved in cell cycle arrest, DNA repair in response to oxidative stress or apoptosis. However, it is unclear how the NO effect on FOXO1. In this study, we observed vital role of FOXO1 dependent transcriptional activation on neuronal death in response to Nitric oxide over-production. We found that NO donor GSNO or L-Arginine significantly increased the FOXO1 transcriptional activity, which induce the FOXO1 downstream proapoptotic genes(Fas L, Bim) expression and finally induce neuronal death. In addition, we screen natural Chinese herb extracts targets to regulating FOXO1 activity. Emodin shows dramatically effect in suppression of FOXO1 transcription activity and protein levels. What's more, Emodin can attenuate neuronal death induced by L-Arg. The current study first demonstrate that Nitric Oxide could regulate FOXO1 transcriptional activity to damage neuronal cell, which will benefit to deep understanding the neurotoxicity of NO free radical. Secondly, by Chinese herb extracts and antioxidants screen, we identified Emodin as one FOXO1 activity modulator which can attenuate neurotoxicity induced by NO. The current study will also provide a new insight for explaining the mechanisms of pathology of neurodegenerative disease and neuroprotective effects of Emodin

AN CLUSTER ALGORITHM FOR THE DATASET WITH MIXED ATTRIBUTES AND APPLICATION TO COMPUTER DYNAMIC FORENSIC

针对目前基于k-MEAnS算法的计算机取证技术存在对符号类型数据处理能力欠缺、误报率较高的问题,提出了一种处理混合型属性的聚类算法的计算机取证技术.该方法将对符号类型特征进行编码映射,并使用主成分分析对编码后增加的维数进行降维,从而解决了在计算机取证中使用聚类分析无法对符号型数据进行处理的问题.文中详细的阐述了改进的具体实现方案,并通过实验验证了该方法的可行性.This paper analyses the exists problems of the current computer dynamic forensic techniques base on K-Means algorithm: can not analysis the feature composed by character,higher false-detection rate,etc,brings forward some improvement.We transform the feature of character to numerical value by mapping,then use the technique of Principal Components Analysis to reduce increased dimensionality after mapping.In this paper,we introduce the improved method concretely,and show the feasibility and effect through an experiment.福建省自然基金项目(2008F50602);福建省自然基金-青年人才项目(2008F3101)的支

Uptake and accumulation of microplastics in a cereal plant wheat

Microplastics pollution is becoming a global environmental concern, and growing evidence has demonstrated the accumulation and distribution of microplastics in terrestrial ecosystems. Once entering into soil, microplastics can change the physical, chemical and biological properties of soil, and then affect the growth of plants. Currently, most attentions have focused on the toxic effects of microplastics on terrestrial plants, only very limited report showed the uptake of microplastics by higher plants under hydroponic culture conditions. The nutrient solution is useful in understanding the mechanism of microplastics uptake, however, it does not account for the importance of affecting factors in the real environment (e.g., the presence of soil organic matter) and therefore do not represent the actual uptake of microplastics in the real-world. Here, we aim to determine whether wheat plants growing in a sand matrix are able to take up 0.2 mum polystyrene (PS) microbeads and translocate these particles from roots to shoots. Wheat was chosen as a representative of cereal crops because it is one of the main staple foods worldwide. A simple and rapid approach for the imaging of fluorescently labelled PS microbeads within plant tissues by confocal laser scanning microscope (CLSM) was used to investigate the uptake, accumulation, translocation and distribution of microspheres in the wheat plant. Two different fluorescent dyes were encapsulated into the PS microbeads matrix and they were used to detect the localization of PS beads in the root and the green tissue respectively. The presence of PS microbeads in plant tissue was then verified using scanning electron microscopy (SEM). Confocal images revealed that the PS luminescence signals were mainly located in the vascular system and on the cell walls of the cortex tissue of the wheat seedling roots after exposure in sand matrix with a concentration of 0.5 g kg~(-1) of PS beads for 21 d, indicated that the beads passed through the intercellular space via the apoplastic transport system. Microbeads clusters were observed in the intercellular space of epidermal tissues and the steles by SEM. Once inside the central cylinder, the 0.2 mum PS beads were transferred from the roots to the stems and leaves via the vascular system. Here, for the first time, we provide evidence of the adherence, uptake, accumulation, and translocation of submicrometer (0.2 mum) PS within the cereal plant in real sand matrix. Our findings provide a methodology and scientific basis for study of the accumulation mechanism of microplastics in soil-crop systems and their potential risk in food chain transfer