ape data were expanded to match with all degrees

lgorithm was used to quantify the damage extent by minimizing the errors between the measured data and numerical results using the two-level search strategy proposed. Numerical results show that the proposed two-level search strategy is often more efficient in quantification of damage extent than the direct search strategy when there are a lot of candidates in the possible damage domain.damage identification; frequencies; genetic algorithm; mode shapes; two-level search strategy教育部留学回国人员科研启动基金资助项目0华中科技大学学报(自然科学版)Journal of Huazhong University of Science and Technology77-79O346.5A004;57;C;AA004_57;程远胜000500030005000773-75基于可靠度的抗震结构优化设     

携带LMO3基因逆转录病毒载体的构建及其在NIH/3 T3细胞中的表达

目的：构建单纯LIM蛋白3（LMO3）的逆转录表达载体,并观察其在 NIH/3T3细胞中的表达情况。方法重组载体pLXSN-LMO3经酶切及测序鉴定后,脂质体法转染到pA317包装细胞,G418筛选稳定的病毒产生细胞株。重组逆转录病毒体外感染NIH/3T3,并对其进行病毒滴度检测,NIH/3T3细胞分为3组：以pLXSN-LMO3转染为实验组,以pLXSN转染为阴性对照组,正常细胞为空白对照组。采用免疫荧光组化染色和蛋白印迹（ Western blot）法鉴定NIH/3T3细胞中LMO3的表达。结果重组逆转录病毒载体pLXSN-LMO3经酶切及测序鉴定构建正确,其病毒滴度平均可达4.04×106 cfu/ml,各组NIH/3T3细胞免疫荧光组化染色及Western blot检测均有LMO3蛋白表达,其中实验组高表达LMO3,与阴性对照组、空白对照组比较差异具有统计学意义（P〈0.05）。结论成功构建了携带LMO3基因逆转录病毒载体,并能在NIH/3T3细胞中高表达LMO3蛋白,为下一步开展基因治疗奠定了基础。     

药物制剂新技术概述及其药剂制作中的实例研究

进入21世纪以来,我国的药物制剂新技术在日新月异的发展。药物制药技术的研究也引起了国内外学者的重视。本文主要介绍了泡腾技术、固体分散技术、纳米技术在我国药物制剂方面的应用。在泡腾技术的实例方面主要从胃内漂浮剂方面、泡腾片方面来研究,指出延长药物吸收时间和提高生物利用度。在固体分散技术主要从聚乙烯吡咯烷酮和乙基纤维素在制药过程中的应用方面展开研究,指出二者催溶性能的优越性。在纳米技术方面主要从抗菌方面、药用气雾剂、聚合物纳米粒等方面来研究。     

地龙的研究进展

地龙是我国常用动物药材,药理作用广泛。笔者从化学成分、药理作用、品种鉴定与分类、养殖加工、品质评价等方面对地龙的研究现状做较全面的综述,指出了各方面研究可能存在的研究空白、重点及难点,为后续地龙相关研究提供参考依据。     

中药穴位敷贴治疗儿童哮喘基础和临床研究现状

从基础研究和临床研究2个方面,包括中医理论研究、实验研究、总体疗效、改善症状、调理体质、疗程时间等,总结了中药穴位敷贴治疗儿童哮喘的研究现状。指出今后研究可设计大样本以便随机对照临床试验研究,同时可在穴位敷贴对儿童中医体质的改善方面开展,以期探讨穴位敷贴的中医作用机制。     

鳖甲化学成分和药理药效研究进展

目的较系统地总结鳖甲在化学成分和药理药效方面的研究情况,为今后进一步深入研究奠定基础。方法查阅近10年来鳖甲的研究资料进行整理和归纳。结果鳖甲的化学成分、药理药效等研究方面有很大进展。结论鳖甲在抗衰老,促进免疫功能,尤其是抗肿瘤方面研究已越来越被重视。     

硒多糖的化学与药理研究进展

硒多糖以其特有的硒和多糖的生物学活性成为近年来研究的热点,本文就国内外近年来对硒多糖的化学方面的提取、分离、纯化、分析和药理学方面的生物学活性、临床应用进行综述,希望对广大从事此方面研究的工作者有所帮助。     

巴西蘑菇抗病毒作用研究动向

在巴西蘑菇抗癌、调节免疫功能等方面已经有不少研究,但在抗病毒方面的研究不多,为探求巴西蘑菇在抗病毒方面是否值得研究．特以国内外发表的文献为材料,介绍巴西蘑菇抗病毒作用的研究动向。可以得出巴西蘑菇具有抗病毒药理作用。并能协同机体免疫而辅助抗病毒,但作用机制还有待进一步研究。     

2009年成瘾研究进展

在美国,每年有超过40万的人死于吸烟、暴饮暴食、物质滥用以及意外事故和自杀。此外,合并有精神疾病的成瘾者（双重诊断）相当普遍。约29％患有精神疾病的人和一半左右患严重精神疾病（如精神分裂症）的人有成瘾史。因此,成瘾相关的预防、研究及治疗依然是精神病学领域关注的重点。我们将回顾2008—2009年在药物滥用、食物和其他成瘾方面的研究。 在药物试验、药物的选择、成瘾易感性、干预的预后以及复吸的可能性方面存在明显的个体差异。在过去的几十年中,在成瘾的认定、预防和治疗方面取得的成绩是有目共睹的。例如,虽然直到DSM—Ⅲ修订版出台才将可卡因列为有滥用潜力的物质,但这一发现拓展了以往局限于对成瘾躯体戒断症状的研究、治疗的范围。最近在动物模型方面已有所突破,目前正在进行有关成瘾大鼠的研究。此外,神经生物学、影像学、蛋白质组学、纳米技术、药理学以及行为科学研究方面的进展也对成瘾领域的研究产生影响。本文综述了我们在成瘾研究中的部分工作以及其他成瘾研究方面取得的进展。     

创新我国中药产品技术标准管理——介绍美国对化妆品监督的特点

经过20世纪的半个多世纪的努力,我国科学工作者在如中药技术标准的研究与制定方面,中药药效物质基础研究、分析方法研究、质量标准研究等方面取得了长足的进步.但是,中药技术标准在继承祖国中医药传统和应用现代技术提高中药产品的质控水平发展方面,已经走到了十字路口上.     

Mechanisms of multimodal sensing by extracellular Ca2+-sensing receptors: a domain-based survey of requirements for binding and signalling

In this article we consider the molecular basis of sensing and signalling by the extracellular calcium-sensing receptor. We consider the nature of its ligands and sensing modalities, the identities of its major protein domains and their roles in sensing, signalling and trafficking as well as the significance of receptor homo- and hetero-dimerization. Finally, we consider the current, incomplete, state of knowledge regarding the requirements for ligand-specific signalling.This article is part of a themed section on Molecular Pharmacology of GPCR. To view the editorial for this themed section visit http://dx.doi.org/10.1111/j.1476-5381.2010.00695.x     

Protein Domain Analysis of C. botulinum Type A Neurotoxin and Its Relationship with Other Botulinum Serotypes

Botulinum neurotoxins (BoNTs) are highly potent poisons produced by seven serotypes of Clostridium botulinum. The mechanism of neurotoxin action is a multistep process which leads to the cleavage of one of three different SNARE proteins essential for synaptic vesicle fusion and transmission of the nerve signals to muscles: synaptobrevin, syntaxin, or SNAP-25. In order to understand the precise mechanism of neurotoxin in a host, the domain structure of the neurotoxin was analyzed among different serotypes of C. botulinum. The results indicate that neurotoxins type A, C, D, E and F contain a coiled-coil domain while types B and type G neurotoxin do not. Interestingly, phylogenetic analysis based on neurotoxin sequences has further confirmed that serotypes B and G are closely related. These results suggest that neurotoxin has multi-domain structure, and coiled-coil domain plays an important role in oligomerisation of the neurotoxin. Domain analysis may help to identify effective antibodies to treat Botulinum toxin intoxication.     

Chemical ligation of unprotected peptides directly from a solid support

In this article we describe a new, convenient procedure to carry out intramolecular (cyclization) and intermolecular native chemical ligations of unprotected peptides directly from a solid support. Our solid-phase ligation approach eliminates the need to manipulate peptide ?thioacid and peptide ^αthioester intermediates in aqueous solution before the ligation step, thereby leading to a reduction in handling losses and significantly increasing the overall efficiency of the chemical ligation strategy. A key step in our ligation scheme is the ability to generate fully unprotected peptides tethered to a solid support through an ”thioester linkage. This can be achieved efficiently using optimized Boc-solid-phase peptide synthesis on a 3-mercaptopropionamide-polyethylene glycol-poly-(N,N-dimethylacrylamide) copolymer support (HS-PEGA). Once the synthesis is complete, the fully protected peptide ?thioester resin is treated with HF to give the corresponding fully unprotected peptide ?thioester resin. Using this procedure several polypeptides ranging from 15 to 47 residues were synthesized successfully. These peptide-resins were then used to perform both intramolecular (head-to-tail cyclizations) and intermolecular solid-phase ligations. The intramolecular solid-phase ligations proceeded much faster than their intermolecular counterparts, but in both cases the reactions were observed to be remarkably clean. The presence of aromatic thiol cofactors significantly accelerated the relatively slow intermolecular ligations. This novel methodology was then extended to provide a general method for performing sequential intermolecular ligations, allowing easy access to much larger polypeptide and protein systems.     

免疫球蛋白结合结构域的研究进展

Z结构域,又叫免疫球蛋白结合结构域,是由金黄色葡萄球菌蛋白A(Staphylococal Protein A,SPA)中B结构域改构而成。SPA是从金黄色葡萄球菌的细胞壁中发现的能与多种哺乳动物的抗体相结合的天然蛋白,组成SPA的5个结构域中,以B结构域的抗体结合力及稳定性占绝对优势。将B结构域28～29位敏感性残基Asn-Gly定向诱变成为Asn-Ala后,得到的结构域被称为Z结构域,其稳定性更高,可耐受羟铵和溴化氢的处理。首尾相连的两个Z结构域,被命名为ZZ结构域,具有与SPA相类似的抗体结合容量以及更高的稳定性,可以耐受工业纯化中苛刻的环境。ZZ结构域以其独特的反3α螺旋结构、以及特有的抗体结合能力已广泛应用于外源蛋白的可溶表达及免疫检测、抗体纯化等领域。该文将对ZZ与抗体结合的机制研究以及它在各领域的应用研究进行综述。     

GST-TAT-P53c融合蛋白的表达及对p53基因突变型肿瘤细胞的促凋亡作用

目的研究P53蛋白C端(P53c)对p53基因突变型肿瘤细胞SW480的促凋亡作用。方法用人类免疫缺陷病毒TAT蛋白的蛋白转导域(TAT)将P53c运载进入肿瘤细胞,用氧依赖性降解区域(ODD)控制P53c在组织中的稳定性。通过PCR方法制备TAT-ODD-p53c(TOPc),TAT-p53c(TPc)及p53c基因,与pGEX4T载体连接后在大肠杆菌中表达谷胱甘肽-S-转移酶(GST)-TAT-ODD-P53c(GST-TOPc),GST-TAT-P53c(GST-TPc)及GST-P53c等融合蛋白。免疫组化方法检测TAT蛋白运载融合蛋白穿过SW480细胞膜的作用,MTT法检测融合蛋白对SW480细胞活力的影响,流式细胞仪检测致SW480细胞凋亡作用。结果成功构建pGEX系列表达载体,并在大肠杆菌中可溶性表达。Western印迹结果表明,重组蛋白可以和GST单克隆抗体特异性结合。免疫组化显示GST-TPc,GST-TOPcm及GST-TOPc均能进入SW480细胞,GST-TPc能明显降低SW480细胞活性,导致SW480细胞凋亡。含ODD的融合蛋白在常氧环境中对肿瘤细胞有轻度促凋亡作用。结论TAT可以介导其融合蛋白跨越细胞膜。GST-TPc可引起p53突变型肿瘤细胞凋亡。     

Heart rate variability after myocardial infarction: what we know and what we still need to find out

Heart rate variability (HRV) is represented by the variation of the time intervals between consecutive heartbeats or the instantaneous heart rates, and can be assessed with linear and non-linear parameters. It is a sensitive indicator of an overall system complexity and adaptability and can be used to diagnose the autonomic dysfunction and quantify the associated risk in a variety of cardiac and non-cardiac disorders. The aim of this review is to summarize the current literature on the value of HRV in predicting the risk of long-term all cause, cardiac, and arrhythmic mortality in survivors of myocardial infarction (MI). We also emphasize the lack of evidence on the role of therapeutic interventions such as medications, bio-behavioral treatments, cardiac rehabilitation, and exercise, in modifying the HRV in post-MI patients.     

脯氨酸羟化酶抑制剂的研究进展

低氧诱导因子(hypoxia-inducible factor,HIF)是细胞适应低氧环境的关键性转录因子,在组织贫(缺)血和炎症性疾病中稳定表达,对缓解病情具有重要作用。脯氨酸羟化酶(proline hydroxylase domain,PHD)能实现HIF的羟基化,是体内降解HIF的最主要蛋白酶。PHD抑制剂可以有效增加体内HIF蛋白含量,对于贫血及炎症性疾病治疗具有积极意义。本研究对小分子PHD抑制剂的最新进展进行综述。     

作用于雄激素受体不同结合位点的前列腺癌治疗药物的研究进展

前列腺癌是最常见的男性泌尿生殖系统的恶性肿瘤。雄激素受体在前列腺癌的发生、发展中起着重要作用。目前,所有治疗前列腺癌的药物(包括第一代的氟他胺、比卡鲁胺、尼鲁米特和第二代的恩扎鲁胺)都与雄激素受体的配体结合口袋结合,并且这些药物有着相似的分子结构,这可能引起药物之间的交叉耐药。为了避免耐药性的产生,研究者们致力于发现雄激素受体上新的药物结合位点。除雄激素受体配体结合位点外,主要对作用于氮端结合位点上的第一活性功能区(AF1)、第二活性功能区(AF2)、AF2附近的第三结合功能区(BF3)和DNA结合位点(DBD)的药物进行综述。     

A negative charge in transmembrane segment 1 of domain II of the cockroach sodium channel is critical for channel gating and action of pyrethroid insecticides

Voltage-gated sodium channels are the primary target of pyrethroids, an important class of synthetic insecticides. Pyrethroids bind to a distinct receptor site on sodium channels and prolong the open state by inhibiting channel deactivation and inactivation. Recent studies have begun to reveal sodium channel residues important for pyrethroid binding. However, how pyrethroid binding leads to inhibition of sodium channel deactivation and inactivation remains elusive. In this study, we show that a negatively charged aspartic acid residue at position 802 (D802) located in the extracellular end of transmembrane segment 1 of domain II (IIS1) is critical for both the action of pyrethroids and the voltage dependence of channel activation. Charge-reversing or -neutralizing substitutions (K, G, or A) of D802 shifted the voltage dependence of activation in the depolarizing direction and reduced channel sensitivity to deltamethrin, a pyrethroid insecticide. The charge-reversing mutation D802K also accelerated open-state deactivation, which may have counteracted the inhibition of sodium channel deactivation by deltamethrin. In contrast, the D802G substitution slowed open-state deactivation, suggesting an additional mechanism for neutralizing the action of deltamethrin. Importantly, Schild analysis showed that D802 is not involved in pyrethroid binding. Thus, we have identified a sodium channel residue that is critical for regulating the action of pyrethroids on the sodium channel without affecting the receptor site of pyrethroids.