无机纳米粒子作为基因载体的研究进展

转染是将具生物功能的核酸转移、运送到细胞内,并使其在细胞内维持生物功能的过程。作为现代生物化学和分子生物学中的一种主要技术手段,转染对于基因治疗有重要的意义。无机纳米粒子作为基因载体受到人们日益广泛的关注,其具有易于制备,可进行多样化的表面修饰等多种优势。本文将概述无机纳米粒子作为基因载体的现状及其对基因表达的影响。     

利用纳米粒子的光学性质高敏感性检测核酸的研究进展

将纳米粒子的光学性质应用于核酸检测是纳米技术应用的一个重要内容,其检测敏感性可达到10-15mol/L(fmol/L,飞摩尔浓度)或亚飞摩尔浓度水平,而且检测成本低、时间短、具有和PCR类似的敏感性、稳定性好,不需要复杂、大型、昂贵的仪器,使得核酸检测可以走出专业的实验室,在野外、家庭中进行。本文主要综述纳米粒子和核酸的偶联,以及利用纳米粒子的光吸收性质、光散射性质、高强度荧光性质进行核酸检测的应用进展。     

生物分子的纳米粒子标记和检测技术

生物分子的标记和检测一直是生物分析领域的重要内容 .近年来 ,纳米材料与生物检测技术的结合 ,使得生物分子的检测有了重要的发展 ,这一交叉学科现已成为生物分析领域最具活力的研究方向 .对近期出现的新型纳米粒子标记物的性质、检测原理、特点和应用进行了评述 ,并分析了用该标记物进行分析的可能发展方向     

脂质纳米粒子在药物中的应用

脂质纳米粒子是用生物可降解的脂质制备,故这种载体系统拥有很好的生物相容性和安全性。本文着重介绍脂质纳米粒子在药物中的应用,如抗肿瘤药物、抗病毒药物、抗炎症药物、免疫药物、抗真菌药物、降血糖药物等。最后,指出了脂质纳米粒子的发展前景。     

应用纳米磁性粒子大幅度提高精制效率超越纳米技术的界限

改良简便且精确地分离、精制、检测样品中微量的蛋白质和病毒等的方法的是生物领域里永恒的课题。     

纳米粒子PCR研究进展

纳米粒子PCR作为纳米技术与生命科学交叉的成果备受关注,对于推动分子生物学技术发展具有重要的意义。详细介绍了纳米粒子PCR的最新研究进展、作用机理,以及纳米粒子PCR应用的具体方面。     

基于超声驱动磁性纳米粒子的肿瘤细胞灭杀

磁性纳米粒子肿瘤热疗技术是目前国际上肿瘤研究的热点.本文提出了一种基于超声驱动磁性纳米粒子(UDMNP)运动进行肿瘤细胞灭杀的新技术,实现磁性纳米粒子的肿瘤治疗.系统研究了肝癌肿瘤细胞HepG2的治疗效果,在一定超声频率下,改变超声功率和超声作用时间,UDMNP具有明显灭杀效果.实验结果显示,较小超声功率下,肿瘤细胞损伤较小,随着超声功率增加,UDMNP对肿瘤细胞表现出明显的灭杀作用.同时,随着作用时间增加,同一超声功率驱动下UDMNP对细胞的灭杀效果也明显提高,光学显微镜观察到细胞形态发生明显变化.本文提出的UDMNP肿瘤细胞灭杀方法的显著优势是减少了化学毒性和有害辐射,是一种物理性机械损伤技术,对促进磁性纳米粒子的临床医学应用有重要意义.     

山楂果实中蛋白质氨基酸及无机元素的分析测定

采用等离子发射光谱法及日立—８３５型氨基酸自动分析仪,分别测定了山楂原种（ＣｒａｔａｅｇｕｓｐｉｎｎａｔｉｆｉｄａＢｇｅ．）的８个单株、山里红（Ｃ．ｐｉｎｎａｔｉｆｉｄａＢｇｅ．ｖａｒ．ｍａｊｏｒＮ．Ｅ．Ｂｒ．）及伏山楂（Ｃ．ｐｒａｅｃｏｘＤａｉ＆Ｆｕ．）果实中的无机元素和蛋白质氨基酸组成含量。结果测得２１种无机元素,其中大量元素６种,微量元素１５种;同时测得蛋白质氨基酸１７种,其中人体必需氨基酸８种。     

蛋白冠与纳米粒子的相互作用

近年来,尽管纳米粒子在生物医学领域的研究中取得了巨大的进展,但很少能进入临床试验阶段,其中,很大程度取决于人们缺乏对纳米粒子与生理环境之间相互作用的认知,对纳米粒子进入体内后的生物学特性了解有限。在生理环境下,蛋白质会吸附于纳米粒子表面,从而形成蛋白冠,这种纳米粒子-蛋白冠复合物的形成严重影响纳米粒子的生物学特性,限制了纳米粒子的临床应用,因此,蛋白质与纳米粒子之间的相互作用应该被深入研究。目前,对纳米粒子-蛋白冠复合物的研究属于一个相对较新的研究领域。概括了蛋白冠的研究现状,对蛋白冠与纳米粒子相互作用所产生的影响进行了重点阐述,也介绍了预防和减少蛋白冠形成的方法,为纳米粒子的进一步研发提供了思路。     

纳米探针用于检测环境中重金属污染物的研究进展

重金属污染对生态环境和人类健康具有极大的危害,建立灵敏、快捷、高效的重金属检测方法具有非常重要的意义.现有的检测技术依赖大型仪器设备,在检测条件、时间以及成本上都有较高的要求,难以满足当前检测工作的需要.随着纳米技术的飞速发展,各种纳米材料不同于块体材料的优异特性被广泛开发,在化学和生物检测领域已有广泛的应用.本文主要综述了近几年来常用的几种纳米探针在重金属检测应用中的研究进展,并对各种纳米探针的特点及检测原理进行了阐述和总结.这些纳米探针包括半导体荧光量子点,荧光纳米粒子、金纳米颗粒等材料,由于他们独特的荧光特性、吸收特性、表面等离子共振(SPR)效应、表面能量转移(SET)效应等,在重金属离子检测领域有很大的应用前景.并且根据目前实际环境监测工作的需要,对基于纳米探针的检测手段进行了讨论和展望,旨在为重金属污染物检测研究的发展和进步提供参考.     

Protein Detection Methods in Proteomics Research

In proteomics research chemical as well as physical methods are used to detect proteins subsequently to their separation. Physical methods are mostly applied after chromatography. They are either based on spectroscopy like light absorption at certain wavelengths or mass determination of peptides and their fragments with mass spectrometry. Chemical methods are used after two-dimensional electrophoresis and employ staining with organic dyes, metal chelates, fluorescent dyes, complexing with silver, or pre-labeling with fluorophores. In some cases autoradiography is still used. Since all of these techniques are very different in terms of sensitivity, their usefulness for quantitative determinations varies significantly. This review will describe the various protein detection methods applied to electrophoresis gels.     

Protein Networks Reveal Detection Bias and Species Consistency When Analysed by Information-Theoretic Methods

We apply our recently developed information-theoretic measures for the characterisation and comparison of protein–protein interaction networks. These measures are used to quantify topological network features via macroscopic statistical properties. Network differences are assessed based on these macroscopic properties as opposed to microscopic overlap, homology information or motif occurrences. We present the results of a large–scale analysis of protein–protein interaction networks. Precise null models are used in our analyses, allowing for reliable interpretation of the results. By quantifying the methodological biases of the experimental data, we can define an information threshold above which networks may be deemed to comprise consistent macroscopic topological properties, despite their small microscopic overlaps. Based on this rationale, data from yeast–two–hybrid methods are sufficiently consistent to allow for intra–species comparisons (between different experiments) and inter–species comparisons, while data from affinity–purification mass–spectrometry methods show large differences even within intra–species comparisons.     

Nanoparticle-based detection technology for DNA analysis

With the current rapid development of nanotechnology and synthesis technology for designed oligonucleotides or oligonucleotide-modified nanoparticle conjugates, the combined strategies have become one of the most valuable methods in detection technology for DNA analysis. Using the uniquely recognizable interactions of pre-designed DNA molecules in assembling nanoparticles, various novel approaches have been recently developed towards detecting specific DNA sequences. Here we describe the key fundamentals and issues of this promising strategies ranging from the initial findings of rationally designed DNA-based assembly of nanoparticles to the extended chip-based detection system. Some limitations of these new strategies and possible approaches will be also discussed for the practical application in the area of DNA microarray detection.     

Detection of Protein Ubiquitination

Ubiquitination, the covalent attachment of the polypeptide ubiquitin to target proteins, is a key posttranslational modification carried out by a set of three enzymes. They include ubiquitin-activating enzyme E1, ubiquitin-conjugating enzyme E2, and ubiquitin ligase E3. Unlike to E1 and E2, E3 ubiquitin ligases display substrate specificity. On the other hand, numerous deubiquitylating enzymes have roles in processing polyubiquitinated proteins. Ubiquitination can result in change of protein stability, cellular localization, and biological activity. Mutations of genes involved in the ubiquitination/deubiquitination pathway or altered ubiquitin system function are associated with many different human diseases such as various types of cancer, neurodegeneration, and metabolic disorders. The detection of altered or normal ubiquitination of target proteins may provide a better understanding on the pathogenesis of these diseases.  Here, we describe protocols to detect protein ubiquitination in cultured cells in vivo and test tubes in vitro. These protocols are also useful to detect other ubiquitin-like small molecule modification such as sumolyation and neddylation.Download video file.^{(96M, mp4)}     

端粒酶活性检测的几种方法

端粒酶活性检测方法的不断发展改进为癌症的诊断治疗以及人们对衰老的进一步研究提供了新途径和新思路。近年来端粒酶活性的检测方法有：(1)基本方法。(2)TRAP法。(3)改良的TRAP法。(4)间接检测法等。     

蛋白质C缺陷及其检测方法

蛋白质C(Protein C，PC)是一种维生素K依赖性的丝氧酸激酶，在止血系统中起重要的调节作用。PC基因异常或一些后天因素可造成PC缺陷，这些因素常共同作用引发静脉血栓。PC总量测定法可用于测定PC总量和PC缺陷分型。PC抗凝活性测定法可测定PC抗凝血活性，其中发色底物法是诊断PC缺陷的首选方法。探讨了实际使用这些方法可能会遇到的一些问题及解决方法。     

Kaede for Detection of Protein Oligomerization

Photoconvertible fluorescent proteins such as Kaede are routinely used for tracking proteins, organelles, and whole cells. Kaede was the first identified photoconvertible fluorescent protein and has since become the most commonly used photoconvertible fluorescent protein in vertebrates. Kaede can be irreversibly converted from a green to a red fluo- rescent form upon UV/blue light irradiation and fluorescence of each form can be isolated separately by appropriate filter sets. Spectral properties of the Kaede forms allow F6rster resonance energy transfer （FRET） from the green form as donor to the red form as acceptor. As a sample containing oligomerized Kaede-containing proteins is exposed to UV or blue light, FRET first increases as green Kaede is converted to red and then decreases as the green donor becomes depleted. Thus, FRET information is potentially obtained from a number of independent measurements taken as photoconversion proceeds. We demonstrate here the application of this approach to detect homo-aggregation and conformational dynamics of plant pro- tein constructs. Structural alterations of 2-cys peroxiredoxin-Kaede were successfully detected depending on the redox state in living plant cells. Photoconversion was performed gradually and donor emission, acceptor emission, and FRET-derived sensitized acceptor emission were measured at each step of conversion. Since photoconvertible proteins have not been rou- tinely used in plants, two plasmids have been designed to facilitate plant applications. The plasmids allow either transient expression of Kaede-containing protein constructs in plant cells or Gateway cloning and stable transformation of plants.