The sirtuin SIRT6 regulates lifespan in male mice

The significant increase in human lifespan during the past century confronts us with great medical challenges. To meet these challenges, the mechanisms that determine healthy ageing must be understood and controlled. Sirtuins are highly conserved deacetylases that have been shown to regulate lifespan in yeast, nematodes and fruitflies. However, the role of sirtuins in regulating worm and fly lifespan has recently become controversial. Moreover, the role of the seven mammalian sirtuins, SIRT1 to SIRT7 (homologues of the yeast sirtuin Sir2), in regulating lifespan is unclear. Here we show that male, but not female, transgenic mice overexpressing Sirt6 (ref. 4) have a significantly longer lifespan than wild-type mice. Gene expression analysis revealed significant differences between male Sirt6-transgenic mice and male wild-type mice: transgenic males displayed lower serum levels of insulin-like growth factor 1 (IGF1), higher levels of IGF-binding protein 1 and altered phosphorylation levels of major components of IGF1 signalling, a key pathway in the regulation of lifespan. This study shows the regulation of mammalian lifespan by a sirtuin family member and has important therapeutic implications for age-related diseases.     

Regulation of cellular response to oncogenic and oxidative stress by Seladin-1

Expression of multiple oncogenes and inactivation of tumour suppressors is required to transform primary mammalian cells into cancer cells. Activated Ha-RasV12 (Ras) is usually associated with cancer, but it also produces paradoxical premature senescence in primary cells by inducing reactive oxygen species followed by accumulation of tumour suppressors p53 and p16(INK4a) (ref. 4). Here we identify, using a direct genetic screen, Seladin-1 (also known as Dhcr24) as a key mediator of Ras-induced senescence. Following oncogenic and oxidative stress, Seladin-1 binds p53 amino terminus and displaces E3 ubiquitin ligase Mdm2 from p53, thus resulting in p53 accumulation. Additionally, Seladin-1 associates with Mdm2 independently of p53, potentially affecting other Mdm2 targets. Ablation of Seladin-1 causes the bypass of Ras-induced senescence in rodent and human fibroblasts, and allows Ras to transform these cells. Wild-type Seladin-1, but not mutants that disrupt its association with either p53 or Mdm2, suppresses the transformed phenotype. The same mutants are also inactive in directing p53-dependent oxidative stress response. These results show an unanticipated role for Seladin-1, previously implicated in Alzheimer's disease and cholesterol metabolism, in integrating cellular response to oncogenic and oxidative stress.     

SRO1 regulates heavy metal mercury stress response in Arabidopsis thaliana

Heavy metals in the environment are harmful limiting factors for the normal growth and development of plants. Here, we isolated and identified an Arabidopsis thaliana T-DNA insertion mutant, named srol-1, which showed a hyper-sensitive response to HgCl2. The SRO1 protein contains a WWE domain that mediates proteinprotein interactions. Under HgCl2 treatment, when compared with the wild-type plants, the growth of srol-1 was repressed dramatically and the number of true leaves was reduced and etiolated. The electrolyte leakage rates showed that cell membrane integrity in srol-1 was damaged more severely than in the wild type. DAB （3,5-diaminobenzidine） staining and confocal microscopy showed that Hg2＋ stress induced more hydrogen peroxide accumulation in srol-1 than in the wild type. The qRT-PCR results indicated that the expression of some abiotic stress-induced genes, such as L-ascorbate peroxidase （APX1）, was reduced under oxidative or Hg2＋ stress. Transgenic plants containing a GFP：：SRO1 fusion protein showed that SRO1 was localized in the nucleus of the cells. SRO1 was shown to be expressed in various tissues, and was most highly expressed in the vigorous tissues. Our results suggest thatSRO1 may play an important role in the stress response of A. thaliana to heavy metals.     

米非司酮对人输卵管胰岛素样生长因子-1及其受体表达的影响

目的: 探讨米非司酮对人输卵管上皮胰岛素样生长因子-1(IGF-1)及其受体(IGF-1R)在增殖中、晚期表达情况的变化.方法:取育龄妇女增殖中、晚期输卵管,米非司酮组(8例)于术前12 h给予米非司酮25 mg口服,对照组(7例)不服用米非司酮,常规石蜡切片,用SP免疫组化法和图像分析技术检测IGF-1及IGF-1R的PU值的表达变化.结果:在月经周期的增殖中、晚期,无论米非司酮组或对照组,峡部、壶腹部、伞部上皮细胞IGF-1及其受体的表达,差异无显著性(P>0.05);在月经周期的增殖中、晚期,米非司酮组和对照组在输卵管相同部位上皮细胞IGF-1的表达差异无显著性(P>0.05);在月经周期的增殖中、晚期,在输卵管相同部位上皮细胞IGF-1R的表达米非司酮组较强,与对照组相比差异有显著性(P<0.05). 结论:米非司酮作用下,月经周期的增殖中、晚期人输卵管上皮IGF-1表达与对照组无明显差异,而IGF-1R的表达则明显强于对照组,说明米非司酮可能对该时期的输卵管环境造成改变.     

好好芭水通道蛋白ScPIP1的功能验证与分析

目的 对转ScPIP1基因烟草植株进行研究以检测ScPIP1的功能及转基因植株的抗逆能力.方法 使用PEG4000对材料进行人工模拟干旱胁迫;使用NaCl对材料进行盐胁迫;使用显微镜对材料叶片气孔形态及指标进行测定.材料的萌发及生长指标使用标尺测量.对所获数据进行统计学分析.结果 Pt2的气孔密度是WT和Pt5的2倍,气孔开度则是WT大于Pt5,Pt5大于Pt2.在不同浓度PEG-4000和NaCl胁迫条件下,Pt2转基因植株的萌发率均大于WT和Pt5.且在胁迫下生长过程中,Pt2的根长与株高均大于WT和Pt5.结论 ScPIP1参与了植物对非生物逆境的应答,并且Pt2是有较高抗逆能力的转基因系.     

IGF-1对绵羊皮肤中GHR、IGF-1、IGF-1R、kAP3.2和KAP6-1基因表达的影响

选取36只健康、体况一致的新疆细毛羊随机分成6组。在每只羊的左侧肩胛部皮下局部注射0．5mL（10ng／mL）的IGF-1,右侧肩胛部皮肤为未注射IGF-1的对照组,然后分别于第0、3、6、9、12、50d采集各皮肤样品,用反转录多聚酶链式反应（RT-PCR）方法,定量分析绵羊皮肤中生长激素受体（GHR）、胰岛素样生长因子1（IGF-1）和胰岛素样生长因子1型受体（IGF-1R）、角蛋白关联蛋白KAP3．2和KAP6-1mRNA的相对丰度。试验结果表明,IGF-1对GHR基因的表达有下调的作用,对IGF-1、IGF-1R基因的表达没有显著的影响,而对KAP3．2、KAP6—1基因的表达具有显著的促进作用。说明IGF-1促进绵羊角蛋白关联蛋白基因的表达可能是通过生长轴以外的其他途径实现的。     

酿酒酵母氧化应激系统的结构生物学基础

酿酒酵母是目前研究背景最为清楚的单细胞真核生物,迄今已知有78个基因编码的蛋白质直接参与其氧化应激反应.这些蛋白质按照功能可以被分为三大类:感应蛋白、调控蛋白和效应蛋白.我们从效应蛋白出发,沿着硫氧还蛋白系统和谷氧还蛋白系统的电子传递路线,逐一解析了所有关键节点蛋白质的三维结构.结合这些蛋白质的生化性质研究、蛋白质-蛋白质复合物的鉴定和结构解析,以及酵母基因组数据库中日益更新的实验数据,我们已初步建立参与酵母氧化应激反应的效应蛋白在原子分辨率上的相互作用网络.这些研究将为我们理解人类氧化应激反应的作用机理提供重要提示,进而可能用于疾病治疗和抗衰老药物的设计.     

Mitochondrial dysfunction and oxidative stress in neurodegenerative diseases

Many lines of evidence suggest that mitochondria have a central role in ageing-related neurodegenerative diseases. Mitochondria are critical regulators of cell death, a key feature of neurodegeneration. Mutations in mitochondrial DNA and oxidative stress both contribute to ageing, which is the greatest risk factor for neurodegenerative diseases. In all major examples of these diseases there is strong evidence that mitochondrial dysfunction occurs early and acts causally in disease pathogenesis. Moreover, an impressive number of disease-specific proteins interact with mitochondria. Thus, therapies targeting basic mitochondrial processes, such as energy metabolism or free-radical generation, or specific interactions of disease-related proteins with mitochondria, hold great promise.     

A sphingosine-1-phosphate receptor regulates cell migration during vertebrate heart development

Coordinated cell migration is essential in many fundamental biological processes including embryonic development, organogenesis, wound healing and the immune response. During organogenesis, groups of cells are directed to specific locations within the embryo. Here we show that the zebrafish miles apart (mil) mutation specifically affects the migration of the heart precursors to the midline. We found that mutant cells transplanted into a wild-type embryo migrate normally and that wild-type cells in a mutant embryo fail to migrate, suggesting that mil may be involved in generating an environment permissive for migration. We isolated mil by positional cloning and show that it encodes a member of the lysosphingolipid G-protein-coupled receptor family. We also show that sphingosine-1-phosphate is a ligand for Mil, and that it activates several downstream signalling events that are not activated by the mutant alleles. These data reveal a new role for lysosphingolipids in regulating cell migration during vertebrate development and provide the first molecular clues into the fusion of the bilateral heart primordia during organogenesis of the heart.     

Type 1 diabetes in mice is linked to the interleukin-1 receptor and Lsh/Ity/Bcg genes on chromosome 1.

Human type 1 (insulin-dependent) diabetes is a common auto-immune disease of the insulin-producing beta cells of the pancreas which is caused by both genetic and environmental factors. Several features of the genetics and immunopathology of diabetes in nonobese diabetic (NOD) mice are shared with the human disease. Of the three diabetes-susceptibility genes, Idd-1 -3 and -4 that have been mapped in mice to date, only in the case of Idd-1 is there any evidence for the identity of the gene product: allelic variation within the murine immune response I-A beta gene and its human homologue HLA-DQB1 correlates with susceptibility, implying that I-A beta is a component of Idd-1. We report here the mapping of Idd-5 to the proximal region of mouse chromosome 1. This region contains at least two candidate susceptibility genes, the interleukin-1 receptor gene and Lsh/Ity/Bcg, which encodes resistance to bacterial and parasitic infections and affects the function of macrophages.     

Post-traumatic stress disorder is associated with PACAP and the PAC1 receptor

Pituitary adenylate cyclase-activating polypeptide (PACAP) is known to broadly regulate the cellular stress response. In contrast, it is unclear if the PACAP-PAC1 receptor pathway has a role in human psychological stress responses, such as post-traumatic stress disorder (PTSD). Here we find, in heavily traumatized subjects, a sex-specific association of PACAP blood levels with fear physiology, PTSD diagnosis and symptoms in females. We examined 44 single nucleotide polymorphisms (SNPs) spanning the PACAP (encoded by ADCYAP1) and PAC1 (encoded by ADCYAP1R1) genes, demonstrating a sex-specific association with PTSD. A single SNP in a putative oestrogen response element within ADCYAP1R1, rs2267735, predicts PTSD diagnosis and symptoms in females only. This SNP also associates with fear discrimination and with ADCYAP1R1 messenger RNA expression in human brain. Methylation of ADCYAP1R1 in peripheral blood is also associated with PTSD. Complementing these human data, ADCYAP1R1 mRNA is induced with fear conditioning or oestrogen replacement in rodent models. These data suggest that perturbations in the PACAP-PAC1 pathway are involved in abnormal stress responses underlying PTSD. These sex-specific effects may occur via oestrogen regulation of ADCYAP1R1. PACAP levels and ADCYAP1R1 SNPs may serve as useful biomarkers to further our mechanistic understanding of PTSD.     

氧化胁迫在TRPV1受体去敏后的活化以及甲醛唤起的痛觉效应中的作用

通过小鼠脚掌注射试验,比较了辣椒素与甲醛在活化TRPV1受体中的直接性与间接性.此外,我们还验证了氧化胁迫对去敏受体的逆转作用,最后验证了甲醛的急性氧化毒性和抗氧化剂对甲醛致痛的抑制作用.该研究证实了氧自由基对TRPV1受体的敏化、活化的正向作用,同时,还证明了这一作用在甲醛活化TRPV1受体介导痛觉过程中的存在性.     

Segregation of receptor and ligand regulates activation of epithelial growth factor receptor

Interactions between ligands and receptors are central to communication between cells and tissues. Human airway epithelia constitutively produce both a ligand, the growth factor heregulin, and its receptors--erbB2, erbB3 and erbB4 (refs 1-3). Although heregulin binding initiates cellular proliferation and differentiation, airway epithelia have a low rate of cell division. This raises the question of how ligand-receptor interactions are controlled in epithelia. Here we show that in differentiated human airway epithelia, heregulin-alpha is present exclusively in the apical membrane and the overlying airway surface liquid, physically separated from erbB2-4, which segregate to the basolateral membrane. This physical arrangement creates a ligand-receptor pair poised for activation whenever epithelial integrity is disrupted. Indeed, immediately following a mechanical injury, heregulin-alpha activates erbB2 in cells at the edge of the wound, and this process hastens restoration of epithelial integrity. Likewise, when epithelial cells are not separated into apical and basolateral membranes ('polarized'), or when tight junctions between adjacent cells are opened, heregulin-alpha activates its receptor. This mechanism of ligand-receptor segregation on either side of epithelial tight junctions may be vital for rapid restoration of integrity following injury, and hence critical for survival. This model also suggests a mechanism for abnormal receptor activation in diseases with increased epithelial permeability.     

高光效水稻品种对氧化胁迫的响应

文章通过对氧化剂甲基紫精处理下高光效水稻杨育粳1号与常规对照品种镇稻11号的鲜重株高的统计、丙二醛含量与叶绿素质量比的测定以及氧化胁迫相关的基因表达分析,研究了水稻的抗氧化机理.研究发现,在20μmol/L甲基紫精处理下,杨育粳1号的鲜重、株高分别是镇稻11号的1.44倍、1.1倍,叶绿素a和b的质量比分别是镇稻11号的2.0倍及2.9倍;活性氧解毒酶mRAN凝胶分析显示,SODA1、APX2及CATB表达量均随甲基紫精浓度增大而增加,且同浓度下前两者在杨育粳1号中的表达量明显高于对照组,据此可推断杨育粳1号抗氧化能力高于镇稻11号.     

AMPK regulates NADPH homeostasis to promote tumour cell survival during energy stress

Overcoming metabolic stress is a critical step for solid tumour growth. However, the underlying mechanisms of cell death and survival under metabolic stress are not well understood. A key signalling pathway involved in metabolic adaptation is the liver kinase B1 (LKB1)-AMP-activated protein kinase (AMPK) pathway. Energy stress conditions that decrease intracellular ATP levels below a certain level promote AMPK activation by LKB1. Previous studies showed that LKB1-deficient or AMPK-deficient cells are resistant to oncogenic transformation and tumorigenesis, possibly because of the function of AMPK in metabolic adaptation. However, the mechanisms by which AMPK promotes metabolic adaptation in tumour cells are not fully understood. Here we show that AMPK activation, during energy stress, prolongs cell survival by redox regulation. Under these conditions, NADPH generation by the pentose phosphate pathway is impaired, but AMPK induces alternative routes to maintain NADPH and inhibit cell death. The inhibition of the acetyl-CoA carboxylases ACC1 and ACC2 by AMPK maintains NADPH levels by decreasing NADPH consumption in fatty-acid synthesis and increasing NADPH generation by means of fatty-acid oxidation. Knockdown of either ACC1 or ACC2 compensates for AMPK activation and facilitates anchorage-independent growth and solid tumour formation in vivo, whereas the activation of ACC1 or ACC2 attenuates these processes. Thus AMPK, in addition to its function in ATP homeostasis, has a key function in NADPH maintenance, which is critical for cancer cell survival under energy stress conditions, such as glucose limitations, anchorage-independent growth and solid tumour formation in vivo.     

Severe impairment of interleukin-1 and Toll-like receptor signalling in mice lacking IRAK-4

Toll-like receptors (TLRs), which recognize pathogen-associated molecular patterns, and members of the pro-inflammatory interleukin-1 receptor (IL-1R) family, share homologies in their cytoplasmic domains called Toll/IL-1R/plant R gene homology (TIR) domains. Intracellular signalling mechanisms mediated by TIRs are similar, with MyD88 (refs 5-8) and TRAF6 (refs 9, 10) having critical roles. Signal transduction between MyD88 and TRAF6 is known to involve the serine-threonine kinase IL-1 receptor-associated kinase 1 (IRAK-1) and two homologous proteins, IRAK-2 (ref. 12) and IRAK-M. However, the physiological functions of the IRAK molecules remain unclear, and gene-targeting studies have shown that IRAK-1 is only partially required for IL-1R and TLR signalling. Here we show by gene-targeting that IRAK-4, an IRAK molecule closely related to the Drosophila Pelle protein, is indispensable for the responses of animals and cultured cells to IL-1 and ligands that stimulate various TLRs. IRAK-4-deficient animals are completely resistant to a lethal dose of lipopolysaccharide (LPS). In addition, animals lacking IRAK-4 are severely impaired in their responses to viral and bacterial challenges. Our results indicate that IRAK-4 has an essential role in innate immunity.     

金属微层裂模拟中的抗拉伸不稳定人为应力方法

金属微层裂是金属在熔化状态下近表面物质的一种拉伸破坏现象,由于该问题的复杂性,一直以来模拟与实验的定量化对比存在难度.本文使用有限元方法模拟了爆轰冲击金属锡微层裂现象,发现微层裂区的模拟出现非物理现象,从而导致自由面速度和微层裂区密度分布均难以比对实验自由面DPS测速及X光结果.针对有限元模拟中出现的微层裂区密度、速度不连续等非物理现象,本文提出了一种抗拉伸不稳定人为应力方法.引入抗拉伸不稳定人为应力后的模拟结果表明:金属微层裂区密度、速度连续性有极大改善,自由面网格脱离现象得到有效抑制,在密度分布及自由面速度上均能够与实验较好符合.通过定量化对比分析认为,虽然抗拉伸不稳定人为应力是并不真实存在的虚假应力,但其作用于金属微层裂稀疏区能够达到稳定计算、减弱或消除非物理现象的作用,具有作用范围限定强、幅值小、不影响原格式守恒性等特点.本文研究突破了金属微层裂模拟与实验难以定量对比的瓶颈问题,为金属微层裂动力学规律分析及物理模型研究提供了重要的模拟技术支撑.