钙蛋白酶及其在内耳的研究进展

钙蛋白酶（calpain）是一类Ca2＋依赖性的半胱氨酸蛋白酶,可参与细胞骨架重构、细胞分化、信号转导、胚胎发育以及调节神经递质释放等多种生理活动。近年来的研究表明,calpain在各种因素引起内耳细胞死亡的病理过程中具有重要的作用。本文就calpain的结构、特性、功能以及在内耳方面的研究进展做一综述。     

钙蛋白酶(Calpain)抑制剂药理活性的研究进展

钙离子(Ca²⁺)作为细胞的第二信使，是参与人体内各项活动的重要调控因子，钙蛋白酶(Calpain),它是一种中性半胱氨酸蛋白酶，主要受Ca²⁺激活，Calpain在许多生理过程中发挥了很大的作用，例如细胞周期的调控与凋亡，还有细胞信号转导和葡萄糖转运等.近年来，人们对这类酶的兴趣大大增加，因为它与脑缺血和其他神经损伤、神经退行性疾病如阿尔茨海默病、心肌缺血相关的神经细胞死亡有很大关系，因此本文对钙蛋白酶抑制剂在神经细胞损伤、失血性休克、缺血再灌注以及阿尔茨海默病研究等方面进行了综述.     

钙蛋白酶在肉嫩化中的作用及其活性测定

介绍了与肉成熟有关的钙激活蛋白酶及其在肉嫩化中的作用、并对钙蛋白酶的活性测定方法、影响酶活性的因素等作了介绍.     

嗜热菌Lon蛋白酶的功能分析

Lon蛋白酶是一种在各种生物体内广泛分布并且具有多种生物功能的蛋白质.以嗜热栖热菌（Thermus thermophilus HB8）的Lon蛋白酶（TTLon）为研究对象,将TTLon蛋白酶的编码基因克隆至pTrc His载体,并在大肠杆菌中进行了成功表达,采用Ni-NTA亲和层析对其进行分离纯化,并通过一系列实验证明Lon蛋白酶能够与ATP结合,具有依赖ATP的蛋白酶活性和依赖ATP的分子伴侣活性.     

玉米巯基蛋白酶抑制剂的纯化及部分性质

从玉米种子纯化的两种巯基蛋白酶抑制剂（ＣＰＩ－Ｉ和ＣＰＩ－Ⅱ）分子量分别为９５００和１３０００,经们都只抑制巯基蛋白酶的水解活性,而对丝氨酸蛋白酶则无抑制作用,属于巯基蛋白酶水解抑制剂,它们在ｐＨ３．０～１１．０或１００℃温度范围内保有１００％的抑制木瓜蛋白的抑制活性,表明在这些外界条件下其分子非常稳定。     

金属蛋白酶临床应用展望

金属蛋白酶是一个大家族,因其需要钙、锌等金属离子作为辅助因子而得名。其家族成员众多,A解联金属蛋白酶（ADAMS）和基质金属蛋白酶（MMPs）等都属于金属蛋白酶超家族。金属蛋白酶在人体内广泛存在,并参与许多重要生理过程,如消化、血管生成和细胞侵润/转移。以往研究主要集中在促进肿瘤侵润转移方面,目前研究表明金属蛋白酶在炎症反应中也有非常重要的作用。     

腈基类化合物对半胱氨酸蛋白酶的抑制机理

半胱氨酸蛋白酶是人体自身产生的重要酶类物质,它广泛的参与人体新陈代谢和蛋白的水解。但过量产生的半胱氨酸蛋白酶会导致骨质疏松和乳腺癌。腈基类化合物分子是一种新发现的具有抑制半胱氨酸蛋白酶作用的靶向药物分子,高效且低副作用。但是它的抑制机理一直没有得到解决。本文介绍借助量子力学/分子力学（Q M/M M）的计算研究,解得了此类化合物对于半胱氨酸蛋白酶的抑制机理,并借助分子学软件设计了一种新型药物分子,有助进一步药物研究。     

雅致放射毛霉AS3.2778碱性蛋白酶的纯化及水解特性

毛霉蛋白酶可以高效地水解大豆蛋白,而且对蛋白水解物具有良好的脱苦效果.为了开发这一蛋白酶系,采用离子交换、疏水层析及凝胶层析等方法,从雅致放射毛霉AS3.2778的发酵麸曲中分离纯化出一蛋白酶组分,并对其水解特性进行了探讨.结果显示：纯化后的蛋白酶是一单体蛋白,其相对分子质量大约为32000;该蛋白酶在55℃、pH8.0-10.0的条件下对大豆蛋白显示出较强的水解能力,其水解效果明显优于商品化的蛋白酶（如木瓜蛋白酶、Alcalase及Protamex）,表明该蛋白酶比以上几种商品化蛋白酶具有更广泛的肽键选择性,对大豆蛋白亲和力更强,在大豆蛋白肽链上有相对较多的酶切位点.     

植物中钙解码机制的研究进展

Ca2＋作为重要的第二信使,在植物生长发育中起着非常重要的作用．目前已经鉴定的植物Ca2＋传感器蛋白有：钙调素（CaM）、类钙调素蛋白（CMLs）、钙依赖型蛋白激酶（CDPKs）和类钙调磷酸酶B亚基蛋白（CBLs）及其互作蛋白激酶（CIPKs）,这些传感器蛋白多以基因家族形式存在,并且在植物中能够形成错综复杂的钙离子信号传递网络系统．主要从植物Ca2＋传感器蛋白的生化特性、生理功能和靶蛋白3个方面,综述了植物解码钙信号机制的最新研究进展．     

基质金属蛋白酶与肿瘤关系研究进展

基质金属蛋白酶是一组Zn2+依赖性蛋白水解酶,能降解细胞外基质,金属蛋白酶抑制剂则能抑制其活性,这两组酶在很多瘤组织中表达增高.文章就基质金属蛋白酶及其抑制剂的表达与肿瘤的关系及在淋巴瘤方面的最新研究进展做了综述.     

Neurotoxicity induces cleavage of p35 to p25 by calpain

Cyclin-dependent kinase 5 (cdk5) and its neuron-specific activator p35 are required for neurite outgrowth and cortical lamination. Proteolytic cleavage of p35 produces p25, which accumulates in the brains of patients with Alzheimer's disease. Conversion of p35 to p25 causes prolonged activation and mislocalization of cdk5. Consequently, the p25/cdk5 kinase hyperphosphorylates tau, disrupts the cytoskeleton and promotes the death (apoptosis) of primary neurons. Here we describe the mechanism of conversion of p35 to p25. In cultured primary cortical neurons, excitotoxins, hypoxic stress and calcium influx induce the production of p25. In fresh brain lysates, addition of calcium can stimulate cleavage of p35 to p25. Specific inhibitors of calpain, a calcium-dependent cysteine protease, effectively inhibit the calcium-induced cleavage of p35. In vitro, calpain directly cleaves p35 to release a fragment with relative molecular mass 25,000. The sequence of the calpain cleavage product corresponds precisely to that of p25. Application of the amyloid beta-peptide A beta(1-42) induces the conversion of p35 to p25 in primary cortical neurons. Furthermore, inhibition of cdk5 or calpain activity reduces cell death in A beta-treated cortical neurons. These observations indicate that cleavage of p35 to p25 by calpain may be involved in the pathogenesis of Alzheimer's disease.     

Expression and proteolytic activity of calpain in lens epithelial cells of oxidative cataract

Objective: To study the role of calpain in the mechanism of oxidative cataract through detecting the level of intracellular free Ca^2 , the expression and proteolytic activity of calpain in the lens epithelial cells (LECs) of H2O2-induced cataract. Methods:Rat lenses were cultured in vitro and cataract was induced by H2O2. The level of intracellular free Ca^2 was measured by fluorescence determination with fura-2/AM. The expression of m-calpain protein in LECs was detected with immunohistochemical method. The proteolytic activity in LECs was measured using a fluorogenic synthetic substrate.Results: There were significant differences of the level of intracellular free Ca^2 (P=0.001, 0.000, 0.000), the expression of m-calpain (P=0.001, 0.000, 0.000) and the proteolytic activity ofcalpain (P=O.O01,0.000, 0.000) between H2O2-induced and control group at 6, 12 and 24 h, respectively. Conclusions: H2O2 can increase intracellular free Ca^2 , then enhance the expression and proteolytic activity of calpain which may play a role in the mechanism of oxidative cataract of rat.     

An essential role for postsynaptic calmodulin and protein kinase activity in long-term potentiation

The phenomenon of long-term potentiation (LTP), a long lasting increase in the strength of synaptic transmission which is due to brief, repetitive activation of excitatory afferent fibres, is one of the most striking examples of synaptic plasticity in the mammalian brain. In the CA1 region of the hippocampus, the induction of LTP requires activation of NMDA (N-methyl-D-aspartate) receptors by synaptically released glutamate with concomitant postsynaptic membrane depolarization. This relieves the voltage-dependent magnesium block of the NMDA-receptor ion channel, allowing calcium to flow into the dendritic spine. Although calcium has been shown to be a necessary trigger for LTP (refs 11, 12), little is known about the immediate biochemical processes that are activated by calcium and are responsible for LTP. The most attractive candidates have been calcium/calmodulin-dependent protein kinase II (CaM-KII) (refs 13-16), protein kinase C (refs 17-19), and the calcium-dependent protease, calpain. Extracellular application of protein kinase inhibitors to the hippocampal slice preparation blocks the induction of LTP (refs 21-23) but it is unclear whether this is due to a pre- and/or postsynaptic action. We have found that intracellular injection into CA1 pyramidal cells of the protein kinase inhibitor H-7, or of the calmodulin antagonist calmidazolium, blocks LTP. Furthermore, LTP is blocked by the injection of synthetic peptides that are potent calmodulin antagonists and inhibit CaM-KII auto- and substrate phosphorylation. These findings demonstrate that in the postsynaptic cell both activation of calmodulin and kinase activity are required for the generation of LTP, and focus further attention on the potential role of CaM-KII in LTP.     

Specific proteolysis of the c-mos proto-oncogene product by calpain on fertilization of Xenopus eggs

The Xenopus c-mos proto-oncogene product, pp39mos, accumulates in the unfertilized egg during maturation, is hyperphosphorylated and exhibits protein kinase activity. On fertilization, or soon after the completion of meiosis, the accumulated pp39mos undergoes selective proteolysis. Using an in vitro protease assay system, we show here that this specific proteolysis is caused by the calcium-dependent cysteine protease, calpain.     

循环流化床锅炉燃烧调整试验研究

锅炉进行燃烧调整是机组投入生产前的一个重要的阶段,通过燃烧调整,对机组各项性能进行综合检验。对东方锅炉(集团)股份有限公司生产的DG1070/17.4-Ⅱ1型锅炉的燃烧调整试验的结果进行了分析总结,指出了该锅炉存在的问题,并提出了相关的建议,同时阐述了燃烧调整过程中主要参数的控制方式及目标值,为日后机组的安全、稳定运行提供了参考。     

A Study on the Molecular Switch of Gene Expression of the Mouse Heart Nuclear DNA Fragments

It is observed by in situ stain that LDH(1-5)…nNAD+ can probably enter the nucleopore and can be bound bound specifically with the genes that encode them. During the in vitro expression, the dilution of heart nuclear DNA fragments could enhance the expression activity of LDH/DNA and the amount of expressed LDH(1-5) is in proportion to the amount of dissociable LDH(1-5) on the LDH/DNA. With the integration of 14CLeu to the proteins, it is also observed that the addition of LDH(1-5)…nNAD+ can suppress the in vitro expression activity of LDH/DNA. AFM bservation shows that the regulation sequence at the both ends of active genes may be bound with such active factors as proteins encoded by the genes which probably is the main molecular switch of gene expression and regulation we have been always searching for. Our work shows the prospective application of the combination of AFM and isotope labeling in the research of biological reaction.     

变结构控制方法在汽车驱动防滑控制中的应用

汽车驱动防滑控制系统是一种新型的主动安全控制技术,通过对滑模变结构控制方法的研究,将其应用于驱动防滑控制中,通过对其进行仿真计算,定性描述了其特性,为汽车驱动防滑控制系统的动力性设计提供了一定的理论依据。     

梭梭根际根瘤菌对紫花苜蓿生长及耐盐性的影响

目的 土壤盐渍化是降低植物生产力、制约农业发展的主要非生物胁迫因素之一,豆科植物共生固氮菌对土壤盐渍化非常敏感,因此探讨盐胁迫下荒漠植物根际根瘤菌资源对紫花苜蓿生长及耐盐性的影响,有望为豆科植物新型复合微生物菌肥的研发和增强紫花苜蓿(Medicago sativa)耐盐能力提供理论基础和优质菌种资源。方法 以从梭梭(Haloxylon ammodendron)根际分离得到的3株根瘤菌(WAW-10、WA30-5和WM30-21)为研究对象,探究0和300 mmol/L NaCl处理下3株梭梭根际根瘤菌对紫花苜蓿生长和耐盐性的影响。结果 接种3株梭梭根际根瘤菌均促进了正常条件和盐胁迫下紫花苜蓿植株的生长,叶绿素含量、根系活力、含碳量和含氮量均显著提升,尤以菌株WM30-21的综合效果最佳;各接菌处理均可诱导紫花苜蓿结瘤,尤以菌株WM30-21的效果最佳。盐胁迫下,接种3株梭梭根际根瘤菌均提高了紫花苜蓿的过氧化氢酶活性,降低了相对质膜透性和丙二醛含量,从而维持了质膜的相对完整性;并提高了紫花苜蓿叶片可溶性糖和脯氨酸含量,从而增强了渗透调节能力;还降低了紫花苜蓿植株中的Na⁺含量、维持K⁺含量的相对稳定,从而提高了钾、钠离子物质量之比(K⁺/Na⁺),其中以菌株WM30-21的效果最佳。结论 接种3株梭梭根际根瘤菌均不同程度地促进了紫花苜蓿的生长,并通过维持质膜的相对完整性、增强渗透调节能力和提高K⁺/Na⁺来增强其耐盐性,其中以菌株WM30-21效果最佳。     

中国频率规范下RFID防碰撞算法性能分析

在射频识别(RFID)系统中,频率规范决定了通信数据率,通信数据率是决定多目标识别性能的重要因素之一.根据我国最新发布的超高频RFID频率规范,通过对信号的频谱仿真分别确定了单读写器环境下,以及标签信号邻道返回时的密集读写器环境下,RFID系统的最大通信数据率,在此通信数据率下对各种符合ISO18000-6C协议的多标签防碰撞算法进行了仿真,结果表明,在不考虑算法在时间和硬件资源上的开销后,Vogt算法在识别超过300个标签时,识别速度(平均每秒识别标签的数量)最高;单读写器环境下最高识别速度约1050个/s,密集读写器环境下邻道返回时的最高识别速度约270个/s.