磷酸化酶

To produce the scientific evidence for developing and manufacturing new antitumor drugs.Methods: 1 The inhibitory effect on cell growth of Hela was measured by MTT assay in treated or untreated groups (3.125, 6.25, 12.5, 25, 50μg/ml TAM and control) for three different treatment times (24h, 48h and 72h).2 Apoptosis and cell cycle were measured by FCM in four experimental groups (0, 4, 16, 40μg/ml TAM) for 48h.3 Adopting Wright and Giemse's staining to observe the morphology of Hela cells which treated with 40μg/ml TAM.4 Using invasion experiment to detect the Hela cells'invasive abilities which treated with 40μg/ml TAM.5 The protein expressional levels of P-ERK, ERK, C-myc and Cyclin D1 in Hela cells untreated or treated with 4, 16, 40μg/ml TAM for 24h were measured by Western blot.6 Expression of anti-apoptotic gene bcl-2, apoptotic gene bax and MMP-9 in Hela cells of four experimental groups (0, 4, 16, 40μg/ml TAM for 24h), were observed by revers transcription PCR.7 The protein expression of P-ERK, ERK, Bcl-2 and Bax in Hela cells treated with 40μg/ml TAM for 24h observed by laser scanning microscopes.

1采用四甲基偶氮唑蓝法检测不同浓度北豆根总碱(3.125、6.25、12.5、25、50μg/ml)处理不同时间(24、48和72小时)对Hela细胞增殖反应的抑制作用。2采用流式细胞技术(flow cytometry，FCM)检测不同浓度北豆根总碱(0、4、16、40μg/ml)作用48小时，对Hela细胞凋亡及周期变化的影响。3瑞氏-姬姆萨染色后显微镜观察北豆根总碱(0、40μg/ml)作用24小时后Hela细胞形态学变化。4采用Transwell小室法检测北豆根总碱(0、40μg/ml)作用24小时后对Hela细胞侵袭性的影响。5采用免疫印迹方法检测不同浓度北豆根总碱(0、4、16、40μg/ml)作用24小时后，Hela细胞中磷酸化ERK、ERK、C-myc、CyclinD1的表达变化。6采用逆转录-聚合酶链反应(revers transcription PCR，RT-PCR)半定量检测北豆根总碱(0、4、16、40μg/ml)作用24小时，Hela细胞抗凋亡基因bcl-2、促凋亡基因bax、基质金属蛋白酶-9(MMP-9)的表达变化。7应用激光共聚焦显微镜(laser scanning microscope，LSM)观察北豆根总碱(0、40μg/ml)作用24小时后，Hela细胞内P-ERK、ERK、Bcl-2、Bax蛋白的表达变化。

METHODS: In this study, we isolated rat pancreatic acini by collagenase digestion; measured the Ca2+/calmodulin-independent activity of autophosphorylated form of the CaM kinase Ⅱ both before and after stimulation of the acini with muscarinic secretagogue bethanechol.

用胶原水解酶法分离大鼠胰腺腺泡，并检测乌拉胆碱刺激腺泡细胞前后自身磷酸化型的CaM kinaseⅡ的活性。

With brain stereotaxis technique PP2A was inhibited in the nucleus basalis of Meynert and the cytoskeletal protein tau, neurofilament were found hyperphosphorylated and aggregating. The neurite became short and thickening, acetyltransferase aggregated in the body of neuron and the acetylcholine level in the nucleus basalis of Meynert was significantly decreased followed with spatial memory defict of rats.

利用立体定位技术抑制大鼠基底核PP2A活性，使基底核神经元微管相关蛋白tau及神经细丝出现过度磷酸化和聚积，轴突变短增粗，胆碱乙酰转移酶在胞体聚集，而使基底核乙酰胆碱含量降低，大鼠出现空间记忆障碍。

Furthermore, phosphorylated ACV directly inhibits HIV-1 reverse transcriptase, terminating DNA chain elongation, and can trap RT at the termination site.

此外，磷酸化的阿昔洛韦能直接抑制HIV-1逆转录酶，将其阻止在终止位点，从而终止DNA链的延长。

To investigate the expression, location and function, long distance PCR was used to expand the targets containing open reading frame. The expressive vectors of prokaryotic and eukaryotic cells were constructed after product purification and connecting with vectors. The prokayotic vectors were induced to express the protein by IPTG and the protein was seen by denaturalization PAG electrophoresis and dyeing. The eukayotic expressive vectors were transfected into culture cells and the protein was found in the nulceolus under the observation of the confocal microscope. The transfected cells were chosen by the geneticin (G418). The cell cyele was examined by flow cytometer and the cell numbers were reduced obviously in the stage of G〓-G〓 and G〓-M, but increased distinguishably in the S stage.

为研究磷酸化应激诱导蛋白的表达、定位和功能，采用了长距离聚合酶链反应扩增含有开放阅读框架的靶片段，经产物纯化、与载体连接，分别构建了STIP1基因的原核表达载体和真核表达载体；IPTG诱导原核表达载体，变性聚丙酰胺凝胶电泳与染色后，可见相应大小的蛋白质表达；STIP1基因的真核表达载体转染细胞系后，共聚焦显微镜下观察STIP1蛋白主要分布于细胞核内；用药物筛选STIP1基因转染的细胞后，流式细胞仪检测细胞周期，可见G〓-G〓期和G〓-M期的细胞明显降低，S期细胞明显增多。

Other polysaccharides are formed from other sugar, which rose by enzymic transformations of phosphorylated hexoes and sugar nucleotides.

其它多聚糖则是由磷酸化己糖和核苷酸糖通过酶作用生成的其它糖聚合形成的。

The protein which act on the phosphated tyrosine receptor kinase is phosphoinsitol 3 kinase , phospholipase C gamma and connective protein Shc, at last the signals can be transferred.

与酪氨酸磷酸化的Trk A相互作用的蛋白质是磷脂酶Cγ和磷脂酰肌醇(-3)激酶(PI3K)，及连接蛋白质Shc，最终传播NGF信号。

Nucleotide-induced eNOS phosphorylation and activity were inhibited by BAPTA-AM (an intracellular free calcium chelator), rottlerin, and protein kinase C siRNA.

BAPTA-AM（一种细胞内游离钙螯合剂），rottlerin和蛋白激酶C siRNA抑制核苷酸介导的内皮型一氧化氮合酶磷酸化和活化。

In conclusion , ERK and telomerase serve a function to some extent in drug resistance of leukemia and ovariancarcinoma cells. The inhibition of ERK signal transduction pathways led to reduction of phosphorylated ERK1 and ERK2protein expression level , and successionally down2regulated the telomerase activity.

通过阻抑ERK通路，降低磷酸化ERK1/ 2蛋白表达水平，进而下调端粒酶活性，可以达到增强白血病和卵巢癌耐药细胞对HRT 或DDP 敏感性的目的。

It will also damage the biomembrane, for example, the damage ofmitochondria will decrease the tricarboxylic acid cycle and oxidative phosphorylation,so the synthesis of ATP will be decresed.

如造成线粒体膜损伤，可减少三羧酸循环和氧化磷酸化，ATP合成障碍；对膜蛋白如酶、受体、离子通道等造成损伤，降低运动能力。

According to the clear water experiment, aeration performance of the new equipment is good with high total oxygen transfer coefficient and oxygen utilization ratio.

曝气设备的动力效率在叶轮转速为120rpm～150rpm时取得最大值，此时氧利用率和充氧能力也具有较高值。

The environmental stability of that world - including its crushing pressures and icy darkness - means that some of its most famous inhabitants have survived for eons as evolutionary throwbacks, their bodies undergoing little change.

稳定的海底环境─包括能把人压扁的压力和冰冷的黑暗─意谓海底某些最知名的栖居生物已以演化返祖的样态活了万世，形体几无变化。