缺磷胁迫导致高等植物叶片中磷脂酰甘油（PG）含量降低的机理研究

磷脂酰甘油（PG）是植物类囊体膜中唯一的磷脂，在它的sn-2位上总是连着一个棕榈酸（16：0）或反式十六碳烯酸（16：1 trans）。由于PG的分子结构独特，对它的功能已有了很多研究，目前认为PG在维持类囊体膜的结构与功能方面具有非常重要的作用。缺磷胁迫下，蓝藻、衣藻及拟南芥、大麦等物种中均检测到了PG含量的下降。对这一现象的常见解释是缺磷导致了PG生物合成受阻，从而引起了其含量的降低。但迄今为止尚没有试验证据支持。本研究比较了缺磷对不同叶龄的小麦与烟草叶片中PG含量与PG水解酶的活性的影响，同时对缺磷叶片酶粗提液水解外源PG后的主要产物、几种磷脂酶抑制剂对上述酶反应的影响等进行了研究，以阐明缺磷条件下叶片中PG含量下降的主要原 因。 缺磷小麦第一叶完全展开时，PG含量与PG水解酶活性均与对照相似;而第三叶完全展开时，尽管缺磷第三叶中PG水解酶活性也与对照相似，但其PG含量低于对照。这一结果表明，在小麦叶片完全展开之前，缺磷条件未影响叶片中的PG水解酶活性，第三叶中较低的PG含量应由PG的生物合成受阻引起。并且，由于缺磷植株第一叶完全展开时PG含量未受影响而第三叶中却表现出了轻微降低，可以推测叶片萌发越晚，PG生物合成受到的抑制就会越严重。 为了研究叶片衰老过程中PG含量下降的原因，我们比较了6，10，14与18日龄时缺磷与对照小麦植株第一叶中PG的相对含量与PG水解酶活性。研究发现：6日龄时，刚刚完全展开的缺磷和对照小麦第一叶中无论是PG含量还是PG水解酶活性都较为相似;而随着叶片的逐渐衰老，缺磷植株第一叶中PG含量大幅度下降，同时伴随着PG水解酶活性的急剧上升。18日龄时，缺磷小麦第一叶中的PG含量较对照降低了69.1%，其PG水解酶活性也远高于对照，37ºC下温育30min后，缺磷叶片的酶粗提液使外源PG含量降低了74.16%，而对照中只降低了13.7%。上述结果表明，缺磷条件下，小麦叶片中PG含量降低的程度与PG水解酶活性的强弱密切相关，PG水解加剧是导致老叶中PG含量降低的一个重要原因。 磷脂酶是水解磷脂的主要酶类。目前在植物体中发现的磷脂酶种类主要有磷脂酶D（PLD）、磷脂酶C（PLC）与磷脂酶A（PLA）。通过薄层层析（TLC），我们发现缺磷小麦叶片的酶粗提液水解外源PG后的主要产物是磷脂酸（PA）、二脂酰甘油（DAG）与游离脂肪酸（FFA）。将n-丁醇加入到缺磷小麦叶片的体外酶反应体系中后，观察到PA、DAG与FFA的生成量均表现出一定程度的降低。由于n-丁醇是PA经PLD途径生成的抑制剂，因此，上述结果表明PLD参与了缺磷条件下小麦叶片中PG的水解。硫酸新霉素是PLC的非特异性抑制剂，低浓度的硫酸新霉素（100μM 和 200μM ）加入到缺磷小麦叶片的体外酶反应体系后，三种产物的生成受到了严重抑制，表明PLC也与缺磷叶片中PG的降解密切相关。 为了进一步分析缺磷导致PG含量降低的原因，我们以烟草为试验材料，检测了缺磷胁迫对烟草嫩叶和老叶中的PG含量、PG水解酶活性、与PG降解相关的酶的种类及PLC、PLDα、PLDβ与PAT-1基因在mRNA上表达水平的的影响。结果表明，缺磷烟草叶片中PG含量的降低由PG生物合成受阻与PG降解加剧共同导致，PLC和PLD活性与烟草叶片中PG的降解有关。缺磷植株老叶中PG水解酶活性及PLC、PLDα、PLDβ基因在mRNA水平上的表达量均高于对照，表明在磷胁迫条件下，老叶中PG水解酶活性可能受到转录水平上的调节， PLC、PLDα、PLDβ转录活性的增强导致了PLC、PLD活性加强，从而引起PG降解的加剧，最终导致了PG含量的降低。与PLC、PLDα和PLDβ不同，缺磷胁迫对patatin蛋白（表现PLA2活性）的编码基因PAT-1在转录水平上的表达无影响，TLC分析PG的水解产物也未检测到溶血磷脂酰甘油（LPG）的生成。由此可见，PLA活性可能与缺磷条件下PG的降解无关。

Bm-TFF2, a trefoil factor protein with platelet activation activity from frog Bombina maxima skin secretions

In mammals, trefoil factor family (TFF) proteins are involved in mucosal maintenance and repair, and they are also implicated in tumor suppression and cancer progression. A novel two domain TFF protein from frog Bombina maxima skin secretions (Bm-TFF2) has been purified and cloned. It activated human platelets in a dose-dependent manner and activation of integrin a(11b)beta(3) was involved. Aspirin and apyrase did not largely reduce platelet response to Bm-TFF2 (a 30% inhibition), indicating that the aggregation is not substantially dependent on ADP and thromboxane A2 autocrine feedback. Elimination of external Ca2+ with EGTA did not influence the platelet aggregation induced by Bm-TFF2, meanwhile a strong calcium signal (cytoplasmic Ca2+ release) was detected, suggesting that activation of phospholipase C (PLC) is involved. Subsequent immunoblotting revealed that, unlike in platelets activated by stejnulxin (a glycoprotein VI agonist), PLC gamma 2 was not phosphorylated in platelets activated by Bm-TFF2. FITC-labeled Bm-TFF2 bound to platelet membranes. Bm-TFF2 is the first TFF protein reported to possess human platelet activation activity. (c) 2005 Elsevier Inc. All rights reserved.

A phospholipase A1 platelet activator from the wasp venom of Vespa magnifica (Smith)

Wasp is an impor tant venomous animal that can induce human fatalities. Aortic thrombosis and cerebral infarction are major clinical symptoms after massive wasp stings but the reason leading to the envenomation manifestation is still not known. In this paper, a toxin protein is purified and characterized by Sephadex G-75 gel filtration, CM-Sephadex C-25 cationic exchange and fast protein liquid chromatography (FPLC) from the venom of the wasp, Vespa magnifica (Smith). This protein, named magnifin, contains phospholipase-like activity and induces platelet aggregation. The cDNA encoding magnifin is cloned from the venom sac cDNA library of the wasp. The predicted protein was deduced from the cDNA with a sequence composed of 337 amino acid residues. Magnifin is very similar to other phospholipase A(1) (PLA(1)), especially to other wasp allergen PLA(1). Magnifin can activate platelet aggregation and induce thrombosis in vivo. The current results proved that PLA(1) in wasp venom could be contributable to aortic thrombosis after massive wasp stings. (c) 2007 Elsevier Ltd. All rights reserved.

Characterization and cloning of a novel phospholipase A(2) from the venom of Trimeresurus jerdonii snake

A phospholipase A(2) (PLA(2)) called jerdoxin, was isolated from Trimeresurus jerdonni snake venom and partially characterized. The protein was purified by three chromatographic steps. SDS-polyacrylamide gel electrophoresis in the presence or absence of dithiothreitol showed that it had a molecular mass of 15 kDa. Jerdoxin had an enzymatic activity of 39.4 mumol/min/mg towards egg yolk phosphatidyl choline (PC). It induced edema in the footpads of mice. In addition, jerdoxin exhibited indirect hemolytic activity. About 97% hemolysis was observed when 2 mug/ml enzyme was incubated for 90 min in the presence of PC and Ca2+. No detectable hemolysis was noticed when PC was not added. Ca2+ was necessary for jerdoxin to exert its hemolytic activity, since only 52% hemolysis was seen when Ca2+ was absent in the reaction mixture. Furthermore, jerdoxin inhibited ADP induced rabbit platelet aggregation and the inhibition was dose dependent with an IC50 of 1.0 muM. The complete amino acid sequence of jerdoxin deduced from cDNA sequence shared high homology with other snake venom PLA(2)s, especially the D49 PLA(2)s. Also, the residues concerned to Ca2+ binding were conserved. This is the first report of cDNA sequence of T jerdonii venom PLA(2). (C) 2002 Elsevier Science Ltd. All rights reserved.

N49 phospholipase A(2), a unique subgroup of snake venom group II phospholipase A(2)

A novel phospholipase A(2) (PLA(2)) with Asn at its site 49 was purified from the snake venom of Protobothrops mucrosquamatus by using SP-Sephadex C25, Superdex 75, Heparin-Sepharose (FF) and HPLC reverse-phage C-18 chromatography and designated as TM-N49

Fluorescence analysis of phospholipase A(2) isolated from Chinese agkistrodon blomhoffii Ussurensis venom

The general and synchronous spectra of phospholipase A(2) (PLA(2)) isolated from Chinese agkistrodon blomhoffii Ussurensis snake venom were studied. The chromophores of PLA(2) were mainly contributed by tyrosine and tryptophane residues when the intervals between the excitation wavelength and the emssion waveleagth (Delta lambda) were 20nm and 75nm, respectively. The pH of buffers could change the fluorescence intensities of PLA(2) by changing the charge distribution of its amino acid chain. Ca2+ can not only increase the emission fluorescence intensity of PLA(2) but also improve the reaction rate of PLA(2) with its corresponding substrate DPPC.

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometric analysis of phospholipase A(2) and fibrinolytic enzyme, two enzymes obtained from Chinese Agkistrodon blomhoffii Ussurensis venom

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) was used to analyze two enzymes, phospholipase AZ and fibrinolytic enzyme isolated from Chinese Agkistrodon blomhoffii Ussurensis venom. Using sinapinic acid as the matrix, positive ion mass spectra of the enzymes were obtained, In addition to the dominant protein [M+H](+) ions, multimeric and multiply charged ions were also observed in the mass spectra, The higher the concentration of the enzymes, the more multiply charged polymer and multimeric ions were detected, Our results indicate that MALDI-TOFMS can provide a rapid and accurate method for molecular weight determination of snake venom enzymes, Mass accuracies of 0.1 and 0.3 % were achieved by analysis of highly dialyzed phospholipase A2 and fibrinolytic enzyme, and these results are much better than those obtained using sodium dodecyl sulfate-palyacrylamide gel electrophoresis. MALDI-TOFMS thus provides a reliable method to determine the purity and molecular weight of these enzymes, which are of potential use as therapeutants, Copyright (C) 1999 John Wiley & Sons, Ltd.

Nonequilibrium microstructures and their evolution in a Fe-Cr-W-Ni-C laser clad coating

The laser-solidified microstructural and compositional characterization and phase evolution during tempering at 963 K were investigated using an analytical transmission electron microscope with energy dispersive X-ray analysis. The cladded alloy, a powder mixture of Fe, Cr, W, Ni, and C with a weight ratio of 10:5:1:1:1, was processed with a 3 kW continuous wave CO2 laser. The processing parameters were 16 mm/s beam scanning speed, 3 mm beam diameter. 2 kW laser power, and 0.3 g/s feed rate. The coating was metallurgically bonded to the substrate, with a maximum thickness of 730 mu m, a microhardness of about 860 Hv and a volumetric dilution ratio of about 6%. Microanalyses revealed that the cladded coating possessed the hypoeutectic microstructure comprising the primary dendritic gamma-austenite and interdendritic eutectic consisted of gamma-austenite and M7C3 carbide. The gamma-austenite was a non-equilibrium phase with extended solid solution of alloying elements and a great deal of defect structures, i.e. a high density of dislocations, twins, and stacking faults existed in gamma phase. During high temperature aging, in situ carbide transformation occurred of M7C3 to M23C6 and M6C. The precipitation of M23C6, MC and M2C carbides from austenite was also observed.

铁基抗高温磨损激光熔覆涂层强韧设计和研究II:Fe-Cr-C-W-Ni激光熔覆层的微观组织及性能

本文以对热工模具进行失效分析的基础上，利用激光熔覆技术，在５ＣｒＭｎＭｏ基础上设计并制备强韧兼备的抗高温磨损涂层。

裂纹顶端位错的运动和小范围屈服的K_(Ⅱc)

本文从裂纹顶端位错运动的简化模型估计形成塑性区所消耗的功;从而计算临界裂纹扩展力。由于Ⅰ型加载和Ⅱ型加载的应力状态有所不同,裂纹顶端塑性功耗也会不一样。基于上述思想,考虑了小范围屈服的K_(Ⅰc)和K_(Ⅱc)的关系,得出K(Ⅱc)>K(Ⅰc)的结论,符合实验结果;并就K(Ⅱc)/K(Ⅰc)的比值与材料性能和温度的关系作了讨论。

铁基抗高温磨损激光熔覆涂层强韧设计和研究 Ⅱ:Fe-Cr-C-W-Ni激光熔覆层的微观组织及性能

利用ＯＭ、ＳＥＭ、ＴＥＭ研究了Ｆｅ－Ｃｒ－Ｃ－Ｗ－Ｎｉ激光熔覆涂层熔覆态及其高温时效态的微观组织结构。结果表明激光熔覆层组织细小，具有强韧两相组成（奥氏体和Ｍ_７Ｃ_３碳化物）的微观结构特征，高温时效处理组织中有Ｍ_（２３）Ｃ_６、Ｍ_６Ｃ、Ｍ_２Ｃ等新碳化物形成。显微硬度和冲击磨损实验证实了激光熔覆态和峰值时效态熔覆层均具有良好的力学性能。

钾离子通道Kir2.1/C-末端功能区的分子动力学模拟

<正>内向整流型钾离子通道(Kir,inwardlyrectifyingpotassium)在细胞激活、细胞内外钾离子K~+的动态平衡、胰岛素分泌等细胞生理过程中起重要作用。而细胞内各种不同因素和第二信使对Kir的调控则是实现其不同生理功能的途径。已有实验结果表明,4,5二磷酸磷脂酰肌醇(PIP2,phosphatidylinositol4,5-bisphosphate)与Kir相互作用的强弱决定了Kir对各种调控因素的响应程度。根据Kir2.1与Kir3.1胞内C-末端X-ray

Visual C++ 6.0学习教程

c="http://img3.douban.com/mpic/s1281493.jpg" border="0" alt="" hspace="8" width="102" height="144" align="left" />MicrosoftVisualC十十6.0作为Microsoft Visual Studio的重要组成部分，包含了迄今为止功能最为强大的基于Windows的应用框架，在同类产品中处于领先地位。VisualC十十6.0是Microsoft迄今为止最全面、最完善的程序开发工具，为了适应各种编程风格，该软件提供了各种各样的辅助工具，在发挥编程能力和提高灵活性方面达到了空前的水平。与以往VisualC十十的各种版本相比较，VisualC十十6.0在编程环境、程序语言技术等方面做了许多改进，从而使VisualC十十更加适合专业程序员快速进行应用程序的开发。

本书内容丰富、图文并茂，是一本适合各种读者学习VisualC十十6.0的优秀参考书。

目 录
第一章 VisualC十十6.0简介及安装
1.1VisualC十十6.0新特性
1.2viSualC十十6.0开发环境简介
1.3如何学习使用VisualC十十6.0
1.4VisualC十十6.0的安装
第二章 走进C十十的世界
2.1类和对象的简介
2.2继承和多态性――一个具体的例子
2.3内嵌对象
2.4在栈中申请对象
2.5全程对象的申请
2.6对象之间的相互关系――指针数据成员
2.7this指针的使用
2.8对指针的引用
2.9友元类和友元函数
2.10静态类成员
2.11重载运算符
2.12从代码中分离出类定义
2.13匈牙利表示法
第三章 VisualC十十6.0的编程环境
3.1VisualC十十6.0主窗口
3.2VisualC十十6.0工具栏
3.3VisualC十十6.0菜单栏
3.4项目与项目工作区
3.5资源与资源编辑器
第四章 编一个最简单的VC十十程序
4.1什么是AppWizard？
4.2迎接你的第一个AppWizard程序
4.3“Iamaprogrammer.”在哪儿？
第五章 程序框架入门
5.1一个简化过的程序框架
5.2WinMain（）：第一个动作
5.3登记窗口类
5.4创建一个窗口
5.5显示窗口
5.6显示出那条消息
5.7窗口类与窗口对象
第六章 消息循环
6.1在消息循环中兜圈子
6.2对事件做出响应：WindowFun（）
6.3响应不同的消息
6.4现在你还跟得上吗？
6.5设备界面进行交互
第七章 精通程序框架
7.1WinMain（）函数在哪儿？
7.2应用程序框架和源文件
7.3工具条、状态条和打印等选项
7.4程序的控制流程
第八章 使用classWizard编程
8.1使用ClassWizard添加消息处理函数
8.2classWizard功能介绍
8.3传送鼠标消息
8.4保存鼠标绘图的信息
第九章 视图与文档
9.1Document－View模式
9.2从视图中分离出文档
9.3保存文档
9.4再访MyProg2.cpp
第十章 对象连接与嵌入（OLE）及其自动化
10.1公共对象模式（COM）
10.2类厂（classfactory）
10.3OLE自动化
10.4IDispatch接口
第十一章 动态连接库（DLLs）
11.1为什么使用DLL
11.2传统的DLL
11.3MFC库DLL
11.4MyProg4A――编写自己的类库扩展DLL
11.5MyProg4B――使用MFC库扩展DLL
11.6资源访问
第十二章 图形设备接口
12.1设备环境类
12.2GDI对象
12.3Windows的颜色映射
12.4映射方式
12.5字体
12.6MyProg3例程序
12.7MyProg3B程序
12.8MyPr0g3C例程序――使用CScrollView
第十三章 对话框
13.1在状态条上显示对话控件的帮助信息
13.2利用Fi1eOpen通用对话框打开多个文件
13.3定制通用文件对话框
13.4扩展和缩减一个对话框
13.5显示一个模式或无模式对话框
13.6编写定制的DDX/DDV例程
第十四章 剖析工具Spy十＋
14.1窗体
14.2消息
14.3进程与线程
第十五章 代码调试
15.1TRACE
15.2调试框架
15.3自我诊断
15.4调试代码的作用
15.5用Dump（）显示对象的信息
15.6检查内存