An enzyme immunoassay for mouse epidermal growth factor utilizing a liquid phase double-antibody system

Summary An enzyme immunoassay for mouse epidermal growth factor (EGF) involving a liquid phase double-antibody system was developed. The EGF--galactosidase conjugate prepared was stable for at least 8 months. By this method, EGF was detectable at a concentration as low as 20 pg per tube. The concentrations of EGF in various tissues of mice are also presented.This work was supported in part by a Grant-in-Aid for Scientific Research from the Ministry of Education, Science and Culture of Japan.     

光交联聚乙烯醇水凝胶及青霉素敏生物传感器研究

合成了一种光交联聚乙烯醇（ＰＶＡ－ＱＱ）水凝胶，着重讨论了该水凝胶固定化β－内酰胺青霉素酶膜及其与离子敏感场效应管（ｐＨ－ＦＥＴ）结合构成的青霉素敏生物传感器的性能。结果表明，ＰＶＡ－ＱＱ水凝胶固定化酶膜具有较高的活性及优良的温度、酸碱度、时间及力学稳定性，由该酶膜制备的青霉素敏ＦＥＴ的响应时间小于６０ｓ，灵敏度大于３ｍＶ／ｍＭ，线性响应范围为０￣１６ｍＭ。     

BP网络的一种优质高效学习算法

对ＢＰ算法的突出问题———收敛性进行分析,综合汇集了五条改进措施,提出一种新的算法．计算机仿真结果表明,该算法对于连续函数,收敛效果较佳;用于正弦函数,迭代１５次即满足误差精度要求Ｅｍａｘ＝１０－３．     

“N1＋的＋N2”短语结构中“的”的隐现规律考察

从认知语法的距离象似性角度来看,“N1＋的＋N2”结构以领属关系居多,领属关系中,N1有的是对N2的限定,有的是N1的部分属性投射到N2。表示限定时,靠近原型领属,结构中的“的”一般会出现,表示属性投射时,接近边缘性领属,“的”一般不会出现。“的”隐现象似于概念距离。     

Rapid reductive degradation of azo dyes by a unique structure of amorphous alloys

An Fe-based amorphous alloy was found to significantly enhance the reductive degradation of an azo dye in aqueous solution. The surface-area normalized rate constant was up to 2.0 L m-2 min-1 at room temperature,an order of magnitude larger than any previously reported value. The activation energy of the degradation process was calculated according to the Arrhenius equation,and a much lower value than those previously reported was obtained. Valence band measurement indicated that a depressed valence band maximum and a widened empty band were established in the amorphous ribbon. The unique structure of the amorphous alloy is important-it may reduce the activation energy of the degradation process and enhance the activity of the electrons,thus accelerating the degradation process.     

Elucidation of the relationship between enzyme activity and internal motion using a lysozyme stabilized by cavity-filling mutations

We investigated the activity and the internal motions of a stabilized mutant hen lysozyme (HEL) in which the residues M12 and L56 were mutated to L and F, respectively (LF mutant HEL). The result of the activity measurements against glycol chitin at various temperatures suggested that the temperature dependence of the activity of LF mutant HEL shifted to the high-temperature side compared with that of wild-type HEL. The detailed internal motions of LF mutant HEL in the absence and presence of a substrate analogue, (NAG)₃, were examined by model-free analysis at 35°C. The results showed that the internal motions of LF mutant HEL in the presence of (NAG)₃ were drastically restricted compared with those in wild-type HEL. Our findings thus suggested that the mutation to the stabilized lysozyme restricted internal motions required for the enzymatic reaction.Received 8 February 2005; accepted 10 March 2005Y. Yoshida and T. Ohkuri contributed equally to this work.     

胰岛素信号通路PI3K／Akt对海马神经元β-淀粉样前体蛋白裂解酶1mRNA水平的影晌

目的通过胰岛素和磷脂酰肌醇-3激酶（P13K）抑制剂渥曼青霉素（wortmannin）对P13K／丝氨酸苏氨酸蛋白激酶（P13K／Akt）信号通路的激活和抑制作用,观察P13K／Akt信号通路对海马神经元β-淀粉样前体蛋白裂解酶1（BACEl）mRNA水平表达的影响。方法20只sD大鼠随机分为空白对照组、假手术组、胰岛素组和渥曼青霉素组,海马立体定向注射胰岛素和P13K抑制剂渥曼青霉素。逆转录一聚合酶链反应（RT-PCR）检测P13K／Akt信号传导下游蛋白Akt以及BACEImRNA水平。结果注射胰岛素的海马P13K信号通路下游信号分子：AktmRNA表达上调（分别较空白和阴性对照组P=0．047,P=0．002）,而BACElmRNA表达下调（分别较空白和阴性对照组P=0．004,P=0．01）。渥曼青霉素组的P13K下游信号分子AktmRNA表达明显被抑制（分别较空白和阴性对照组P=0．002,P=0．039）,同时BACEImRNA的表达较对照组上调（分别较空白和阴性对照组P=0．039,P=0．018）。结论胰岛素信号通路P13K／AM可以调节BACEl的转录水平参与阿尔茨海默病的发病机制。     

外源性H2S通过调节β-位淀粉样前体蛋白裂解酶1表达对PC12细胞APP／Aβ代谢的影响

目的观察外源性硫化氢对嗜铬细胞瘤细胞β-位淀粉样前体蛋白裂解酶1（BACE1）的调节作用,进而探讨其对淀粉样前体蛋白／β-位淀粉样蛋白代谢途径的影响。方法用硫氢化钠作外源性H2s供体,实验设空白对照组、NaHs50μmol／L组、NaHS100μmol／L组和NariS200μmol／L组,按分组浓度处理PC12细胞24h后,RT-PCR和Western blot法检测细胞内BACE1 mRNA及蛋白表达,并用Western blot法继而检测APP代谢过程中关键蛋白APP、C99、C83表达变化,EusA法检测细胞培养液中AB40和AB42水平。结果NaHS在实验浓度范围内从基因与蛋白两个水平上呈剂量依赖性下调BACE1表达,并下调C99、Ap40和Ap42蛋白表达,上调C83蛋白,各NaHS组分别与对照组比较,差别均有统计学意义（P〈0．05）,而对APP蛋白表达没有影响,各组间比较差别无显著性（P〉0．05）。结论外源性H2s具有通过调节PC12细胞BACE1表达下调APP／Aβ代谢的作用。     

肥胖以及有氧耐力运动对大鼠血清抗氧化酶和一氧化氮的影响

研究了肥胖以及耐力性运动对大鼠血清抗氧化酶和一氧化氮水平的影响,结果表明,高脂饮食组实验大鼠血清中SOD和CAT浓度显著性下降,MDA明显增加,NO活性下降;高脂饮食结合有氧耐力运动组大鼠血清中SOD和CAT活性以及MDA明显上升,血清中NO水平恢复明显.肥胖大鼠体重增长被明显抑制.结合生化试验结果,确定高脂饮食诱发肥胖后可造成实验动物血清中抗氧化酶活性下降,自由基生成增加,血管舒张功能下降,系统的有氧耐力运动可以有效的逆转高脂饮食造成的自由基增加,NO恢复至正常水平.