An intracellular P2X receptor required for osmoregulation in Dictyostelium discoideum

P2X receptors are membrane ion channels gated by extracellular ATP that are found widely in vertebrates, but not previously in microbes. Here we identify a weakly related gene in the genome of the social amoeba Dictyostelium discoideum, and show, with the use of heterologous expression in human embryonic kidney cells, that it encodes a membrane ion channel activated by ATP (30-100 muM). Site-directed mutagenesis revealed essential conservation of structure-function relations with P2X receptors of higher organisms. The receptor was insensitive to the usual P2X antagonists but was blocked by nanomolar concentrations of Cu2+ ions. In D. discoideum, the receptor was found on intracellular membranes, with prominent localization to an osmoregulatory organelle, the contractile vacuole. Targeted disruption of the gene in D. discoideum resulted in cells that were unable to regulate cell volume in hypotonic conditions. Cell swelling in these mutant cells was accompanied by a marked inhibition of contractile vacuole emptying. These findings demonstrate a new functional role for P2X receptors on intracellular organelles, in this case in osmoregulation.     

Extracellular sodium regulates airway ciliary motility by inhibiting a P2X receptor.

The mucociliary system is responsible for clearing inhaled particles and pathogens from the airways. This important task is performed by the beating of cilia and the consequent movement of mucus from the lungs to the upper airways. Because ciliary motility is enhanced by elevated intracellular calcium concentrations, inhibition of calcium influx could lead to disease by jeopardizing mucociliary clearance. Several hormones and neurotransmitters stimulate ciliary motility, one of the most potent of which is extracellular ATP (ATP0), which acts by releasing calcium ions from internal stores and by activating calcium influx. Here we show that, in airway ciliated cells, extracellular sodium ions (Na+(0)) specifically and competitively inhibit an ATP0-gated channel that is permeable to calcium ions, and thereby attenuate ATP0-induced ciliary motility. Our finding points to a physiological role for Na+(0) in ciliary function, and indicates that mucociliary clearance might be improved in respiratory disorders such as chronic bronchitis and cystic fibrosis by decreasing the sodium concentration of the airway surface fluid in which the cilia are bathed.     

Establishment of a high-throughput screening system for universal anti-HIV targets

The process of identifying novel human immunodeficiency virus 1 (HIV-1) inhibitors presents a challenge for industrial and scientific research.Virus-cell-based screening approaches offer some advantages in the quest for novel inhibitors because they include multiple targets in a single screen and in some cases reveal targets not captured in biochemical assays.In this study,a high-throughput screening (HTS) system for HIV-1 inhibitors was developed,which allows the simultaneous screening of all the HIV-1 tar...     

P2X receptors as cell-surface ATP sensors in health and disease

P2X receptors are membrane ion channels activated by the binding of extracellular adenosine triphosphate (ATP). For years their functional significance was consigned to distant regions of the autonomic nervous system, but recent work indicates several further key roles, such as afferent signalling, chronic pain, and in autocrine loops of endothelial and epithelial cells. P2X receptors have a molecular architecture distinct from other ion channel protein families, and have several unique functional properties.     

Molecular mechanism of ATP binding and ion channel activation in P2X receptors

P2X receptors are trimeric ATP-activated ion channels permeable to Na+, K+ and Ca2+. The seven P2X receptor subtypes are implicated in physiological processes that include modulation of synaptic transmission, contraction of smooth muscle, secretion of chemical transmitters and regulation of immune responses. Despite the importance of P2X receptors in cellular physiology, the three-dimensional composition of the ATP-binding site, the structural mechanism of ATP-dependent ion channel gating and the architecture of the open ion channel pore are unknown. Here we report the crystal structure of the zebrafish P2X4 receptor in complex with ATP and a new structure of the apo receptor. The agonist-bound structure reveals a previously unseen ATP-binding motif and an open ion channel pore. ATP binding induces cleft closure of the nucleotide-binding pocket, flexing of the lower body β-sheet and a radial expansion of the extracellular vestibule. The structural widening of the extracellular vestibule is directly coupled to the opening of the ion channel pore by way of an iris-like expansion of the transmembrane helices. The structural delineation of the ATP-binding site and the ion channel pore, together with the conformational changes associated with ion channel gating, will stimulate development of new pharmacological agents.     

关于丢番图方程x3-1=7y2的初等解法

关于丢番图方程x3-1=7y2的初等解法的问题至今仍未得到解决.主要运用递归序列、同余式方法、Maple小程序和二次剩余初等方法以及Pell方程的解的性质,证明了此方程仅有整数解(x,y)=(1,0),(2,±1),(4,±3),(22,±39).     

基态7Li2(X1Σ+g)分子的解析势能函数

利用分子反应静力学的原理,导出了7L i2分子X1Σg 态的合理离解极限;使用HF、B3LYP、B3P86、QC ISD和QC ISD(T)方法和D95、D95V、D95V(d,p)、D95V(3df,3pd)、6-311G、6-311G(d)、6-311G(d,p)、6-311G(3df,3pd)基组,对7L i2分子X1Σg 态的平衡几何、谐振频率和离解能进行了优化计算,比较得出QC ISD(T)/6-311G(3df,3pd)为最优方法/基组.在QC ISD(T)/6-311G(3df,3pd)水平下,对7L i2分子X1Σg 态进行了单点能扫描,并用正规方程组拟合出了其解析势能函数.根据拟合出的解析势能函数计算出了X1Σg 态的光谱常数Be=0.656 cm-1,αe=0.006 cm-1和ωeeχ=2.85 cm-1及二阶、三阶和四阶力常数f2=26.159 aJ.nm-2,f3=-533.479 aJ.nm-3和f4=5 688.5 aJ.nm-4.     

Urinary bladder hyporeflexia and reduced pain-related behaviour in P2X3-deficient mice

Extracellular ATP is implicated in numerous sensory processes ranging from the response to pain to the regulation of motility in visceral organs. The ATP receptor P2X3 is selectively expressed on small diameter sensory neurons, supporting this hypothesis. Here we show that mice deficient in P2X3 lose the rapidly desensitizing ATP-induced currents in dorsal root ganglion neurons. P2X3 deficiency also causes a reduction in the sustained ATP-induced currents in nodose ganglion neurons. P2X3-null mice have reduced pain-related behaviour in response to injection of ATP and formalin. Significantly, P2X3-null mice exhibit a marked urinary bladder hyporeflexia, characterized by decreased voiding frequency and increased bladder capacity, but normal bladder pressures. Immunohistochemical studies localize P2X3 to nerve fibres innervating the urinary bladder of wild-type mice, and show that loss of P2X3 does not alter sensory neuron innervation density. Thus, P2X3 is critical for peripheral pain responses and afferent pathways controlling urinary bladder volume reflexes. Antagonists to P2X3 may therefore have therapeutic potential in the treatment of disorders of urine storage and voiding such as overactive bladder.     

Warm-coding deficits and aberrant inflammatory pain in mice lacking P2X3 receptors

ATP activates damage-sensing neurons (nociceptors) and can evoke a sensation of pain. The ATP receptor P2X3 is selectively expressed by nociceptors and is one of seven ATP-gated, cation-selective ion channels. Here we demonstrate that ablation of the P2X3 gene results in the loss of rapidly desensitizing ATP-gated cation currents in dorsal root ganglion neurons, and that the responses of nodose ganglion neurons to ATP show altered kinetics and pharmacology resulting from the loss of expression of P2X(2/3) heteromultimers. Null mutants have normal sensorimotor function. Behavioural responses to noxious mechanical and thermal stimuli are also normal, although formalin-induced pain behaviour is reduced. In contrast, deletion of the P2X3 receptor causes enhanced thermal hyperalgesia in chronic inflammation. Notably, although dorsal-horn neuronal responses to mechanical and noxious heat application are normal, P2X3-null mice are unable to code the intensity of non-noxious 'warming' stimuli.     

Oracle7.X数据库故障的恢复

介绍了Ｏｒａｃｌｅ数据库故障类型、故障恢复方法及故障恢复所使用的数据结构，给出了两个故障恢复实例，为Ｏｒａｃｌｅ数据库系统管理损恢复故障提供有益的经验和技巧。     

面向客户关系的X2X商务模式研究

文章回顾了商务模式的发展历程，并对现有电子商务模式常见的划分进行了列举和对比，并基于客户关系的角度提出了面向客户关系的电子商务X2X综合模式，最后对于电子商务的模式选择与革新从面向客户的角度给出了一些指导原则。     

CH2(X3B1)+O2→CH2O+O(3P)自由基反应机理理论研究

采用相关能校正自洽场分子轨道MP2/6-311G**从头计算法,结合能量梯度法研究了CH2(X3B1)+O2→CH2O+O的反应机理,优化了反应势能面上反应物、过渡态、中间体和产物的几何构型,并采用内禀反应坐标理论(IRC)计算了该反应的反应途径,得出该反应为一历经中间体的分步反应,支持了实验工作者提出的机理.     

Reduced vas deferens contraction and male infertility in mice lacking P2X1 receptors

P2X1 receptors for ATP are ligand-gated cation channels, present on many excitable cells including vas deferens smooth muscle cells. A substantial component of the contractile response of the vas deferens to sympathetic nerve stimulation, which propels sperm into the ejaculate, is mediated through P2X receptors. Here we show that male fertility is reduced by approximately 90% in mice with a targeted deletion of the P2X1 receptor gene. Male mice copulate normally--reduced fertility results from a reduction of sperm in the ejaculate and not from sperm dysfunction. Female mice and heterozygote mice are unaffected. In P2X1-receptor-deficient mice, contraction of the vas deferens to sympathetic nerve stimulation is reduced by up to 60% and responses to P2X receptor agonists are abolished. These results show that P2X1 receptors are essential for normal male reproductive function and suggest that the development of selective P2X1 receptor antagonists may provide an effective non-hormonal male contraceptive pill. Also, agents that potentiate the actions of ATP at P2X1 receptors may be useful in the treatment of male infertility.     

P2X4 receptors induced in spinal microglia gate tactile allodynia after nerve injury

Pain after nerve damage is an expression of pathological operation of the nervous system, one hallmark of which is tactile allodynia-pain hypersensitivity evoked by innocuous stimuli. Effective therapy for this pain is lacking, and the underlying mechanisms are poorly understood. Here we report that pharmacological blockade of spinal P2X4 receptors (P2X4Rs), a subtype of ionotropic ATP receptor, reversed tactile allodynia caused by peripheral nerve injury without affecting acute pain behaviours in naive animals. After nerve injury, P2X4R expression increased strikingly in the ipsilateral spinal cord, and P2X4Rs were induced in hyperactive microglia but not in neurons or astrocytes. Intraspinal administration of P2X4R antisense oligodeoxynucleotide decreased the induction of P2X4Rs and suppressed tactile allodynia after nerve injury. Conversely, intraspinal administration of microglia in which P2X4Rs had been induced and stimulated, produced tactile allodynia in naive rats. Taken together, our results demonstrate that activation of P2X4Rs in hyperactive microglia is necessary for tactile allodynia after nerve injury and is sufficient to produce tactile allodynia in normal animals. Thus, blocking P2X4Rs in microglia might be a new therapeutic strategy for pain induced by nerve injury.     

矩阵方程X+A~*X~2A=P的Hermite正定解及扰动分析

考虑非线性矩阵方程X＋A·X^2A=P,其中A是一个n×n阶的复矩阵,P是一个n×n阶的Hermite正定矩阵,A＊表示矩阵A的共轭转置。推导出矩阵方程的Hermite解的存在及唯一性条件,同时给出唯一解的存在区间。最后对该唯一解进行扰动分析,给出不依赖于扰动解的扰动边界。     

Fe_2P_2O_7制备锂离子电池正极材料LiFePO_4的碳包覆优化研究

利用Fe_2P_2O_7和碳酸锂为原材料,并通过不同的碳包覆合成LiFePO_4/C复合材料.利用XRD、SEM、碳硫分析仪、恒流充放电法和循环伏安对产物的组成、结构、形貌和电化学性能进行测试,确定含碳量为2.45wt%的LiFePO_4/C复合材料具有更好的电化学性能.实验结果表明,在0.1C倍率下,锂离子电池的放电比容量为130.49 m Ah/g,在1C倍率下,锂离子电池的放电比容量为108.58 m Ah/g.     

4-氯-7-硝基苯并噁二唑的合成及对次氯酸根的荧光检测

以2,6-二氯苯胺为原料合成了4-氯-7-硝基苯并噁二唑的荧光探针,其结构经过NMR进行了表征。利用荧光光谱探讨了该探针对次氯酸根的荧光响应性能。实验结果表明,该探针对次氯酸根具有良好的选择性和灵敏性。在V_(CH_3CH_2OH):V_(PBS)=1:1(50 m M PBS buffer,pH7.4)的缓冲溶液中,探针随着次氯酸根浓度的增加而增强,探针对次氯酸浓度在0~16μM范围内具有较好的线性关系,其对次氯酸根的检测下限为0.23μmol/L。     

P2P软件中UPnP功能的实现

主要研究UPnP在P2P软件中的应用及UPnP功能实现的原理和步骤.因为现在互联网有一部分机器是工作在内网,而UPnP应用于P2P软件中可以将软件性能提高30%左右,利用UPnP实现端口映射对P2P软件性能提高有大?篌帮助.