基于电化学技术的microRNA生物传感器

MicroRNA(miRNA)是一种内源性的非编码单链RNA,通过与mRNA的3'端非翻译区(UTR)的不完全互补或完全互补结合抑制靶mRNA的翻译或促使靶mRNA的降解来调控基因的表达,参与细胞的增殖、凋亡、分化和代谢等重要过程。MiRNA表达的变化可以起到癌基因和抑癌基因的作用,是一种潜在的肿瘤标志物,因此,miRNA的检测技术引起了人们的关注。由于电化学检测方法具有灵敏、快速、低成本和低能耗等特点,研究者广泛开展了应用电化学技术来发展miRNA检测的研究。本文将对基于电化学技术的miRNA检测方法进行综述。     

超灵敏检测miRNA

最新研究表明MicroRNAs(miRNA)在细胞内的异常表达与人类疾病具有密切的关系,并且miRNA有望成为一种肿瘤标志物和治疗靶点。miRNA在细胞增殖、分化和凋亡等生物学过程中起着重要作用。最新研究表明miRNA在细胞内的异常表达与人类疾病具有密切的关系,并且miRNA有望成为一种肿瘤标志物和治疗靶点。传统的Northern杂交法和实时定量PCR技术常被用于miRNA的分析,但其灵敏度低与特异性差等局限性限制了这两种方法的广泛应用。因     

MicroRNA体外检测的研究进展

MicroRNA (miRNA)是一类内源性、进化高度保守的小分子非编码RNA,通过识别同源序列及干扰转录、翻译或表观遗传以调节基因的表达。研究发现,某些miRNA的异常表达与疾病相关,可作为生物标志物或药物靶点为疾病诊断、治疗及预后提供新思路,而准确测定miRNA的表达是其应用于临床的关键。本文结合近年来研究成果对传统检测方法及其改进和等温核酸扩增的新技术进行概述,分析这些方法的优势与不足。     

基于DNA “Y”型结构的指数型核酸恒温扩增策略检测microRNA

该文基于“Y”型DNA模板和恒温指数扩增反应(EXPAR)技术的信号放大功能,建立了一种快速低序列依赖性的miRNA检测方法(Y-EXPAR)。以let-7b为检测目标,设计两条部分互补的DNA模板,与目标miRNA共同构成“Y”型结构,能够同时进行双向扩增,反应效率高,反应时间短,并降低了放大性能对模板序列的依赖。方法检出限低至0.3 amol/L(相当于10 μL溶液中有1.8拷贝的let-7b),并能区分单碱基差异的let-7家族同源序列。应用于肿瘤细胞裂解液let-7b的检测,POI值与细胞数目对数值呈线性关系,说明Y-EXPAR在实际样品miRNA的检测中具有优势。得益于扩增的高特异性、抗干扰性和低序列依赖性,该方法为miRNA的快速检测提供了新思路,在miRNA相关的疾病研究和临床诊断方面具有良好应用前景。     

纳米孔道单分子检测结直肠癌MicroRNAs

MicroRNA（miRNA）可用于癌症的早期诊断、预后判断,其分析检测具有重要临床意义.结直肠癌的发生、发展与miRNA 21、miRNA 92等的异常表达明显相关.本研究设计了以poly（dT）_n为引导链的DNA探针（probe）并尝试使用α-溶血素（α-HL）纳米孔道单分子检测方法检测结直肠癌miRNAs.miRNA·probe复合物分子穿过α-HL纳米孔道限域空间时,由于probe链长、序列不同导致probe-α-HL相互作用不同,miRNA 92·probe 92、miRNA 21·probe 21、miRNA 16·probe 16输出为形态、阻断时间不同的多台阶特征信号,实现了三种miRNAs的有效区分.实验证明,此方法可以用于检测血清实际样品.因此,未来有望使用α-HL构建miRNA超灵敏单分子生物传感器.     

RNA生物标志物体外检测方法研究进展

RNA生物标志物主要包括编码蛋白的mRNA和非编码蛋白的microRNA (miRNA)、环状RNA (circula rRNA, circRNA)及长链非编码RNA (lncRNA)等.RNA生物标志物种类多,生物信息量丰富,相比DNA更能动态反映细胞生物学功能和调控过程,因此,RNA生物标志物的定量检测和表达分析对于细胞功能研究、疾病的诊断和治疗及预后监测等具有重要意义.本文重点介绍了常见RNA生物标志物mRNA、miRNA、circRNA和lncRNA体外检测方法的原理及研究进展,并对RNA生物标志物检测面临的挑战及发展趋势进行了展望.     

MicroRNAs电化学生物传感器在乳腺癌检测中的研究进展

MicroRNAs是一类内源性非编码小RNA分子,可调控靶基因的表达。特异性microRNAs的失调在诸如癌症、心血管疾病、免疫疾病、神经退行性疾病和皮肤疾病等的发展过程中起着关键作用,常作为疾病早期诊断和预后的生物标志物。电化学生物传感器由于其灵敏、快速、成本低等优势,已经成为传统microRNAs检测方法的一种很有前途的替代方法。本文综述了基于microRNA电化学生物传感器检测乳腺癌的研究进展,并对其所构建的电化学生物传感器的检测方法进行了分析探讨。     

aprE基因表达的分子生物学和微量热法分析

利用分子生物学方法(SDS-PAGE和酶活检测法)未检测到所克隆的aprE基因在大肠杆菌中的表达产物(碱性蛋白酶), 而微量热法检测结果发现: 重组菌株的生长代谢产热曲线之间存在明显的差异. 根据这些差异, 分析了该基因的上、下游调控序列对该基因在大肠杆菌中表达的重要作用, 从而进一步对该基因进行了亚克隆, 得到了生物学方法可检测到的表达产物. 由此推测, 微量热技术有可能为检测外源基因表达及其调控, 以及为指导进一步基因工程操作提供一种新的快速灵敏的技术和方法.     

基于流式微球分析技术的超高灵敏度蛋白激酶活性分析

蛋白激酶引发的蛋白质磷酸化在细胞信号转导过程中发挥着极其重要的调控作用。研究表明,众多的人类疾病和蛋白激酶的活性异常密切相关,蛋白激酶也因而成为治疗癌症、炎症和其它免疫调控紊乱等复杂性疾病的重要药物靶点,开发、筛选小分子蛋白激酶活性抑制剂是当前新药研发的重要方向。因此,建立操作简单、成本低廉、灵敏度高的蛋白激酶活性检测方法是实现以蛋白激酶为靶点进行疾病诊断及新药开发的先决条件。传统激酶活性分析方法主要是依赖放射性32P标记或对特定磷酸化     

硒蛋白的分子生物学及与疾病的关系*

硒蛋白是微量元素硒在体内存在和发挥生物功能的主要形式。因硒蛋白的活性中心硒代半胱氨酸由传统终止码TGA编码,故从基因组中预测硒蛋白以及用基因工程技术表达硒蛋白均很困难。有关硒抗氧化、对癌症、神经退行性疾病和病毒作用的报导较多,但结论并不一致。本文综述了硒蛋白基因预测、蛋白质表达调控以及硒和硒蛋白对癌症、神经退行性疾病和病毒的作用及机制等方面的近期进展,研究提高硒蛋白生物信息学预测准确率和基因工程表达量的方法,分析了解硒蛋白与疾病发生发展的关系和机制,探索不同硒蛋白作为预防药物开发、作为癌症治疗和药物筛选靶标的可能性。     

二氧化碳和丙烯直接合成甲基丙烯酸的NiPMo/TiO2催化剂的研究

用溶胶-凝胶法以磷钼酸(MPA)的镍盐溶液水解钛酸四丁酯制备了NiPMo/TiO2催化剂.使用ICP、 XRD、 TG-DTA、 IR、 TPD-MS和微反应技术研究了催化剂的化学组成、热稳定性、化学吸附性质和催化反应性能.杂多钼酸盐与TiO2通过O2-在TiO2表面发生了键合.在623 K下,杂多阴离子仍保持原有的Keggin结构.CO2在Lewis酸位Ni(Ⅱ)和Lewis碱位Ni-O-Mo的桥氧协同作用下生成CO2卧式吸附态Ni(Ⅱ)←O-(CO)←(O--Ni).丙烯有多种吸附态在催化剂上吸附.在563 K、 1 MPa和空速1500 h-1的反应条件下,丙烯的摩尔转化率为3.2%,产物MAA选择性为95%.     

Toward new camptothecins. Part 6: Synthesis of crucial ketones and their use in Friedländer reaction

In the context of the preparation of camptothecin and luotonin A analogs, the synthesis of some key keto-precursors and their use in Friedländer condensation are described. This paper also focuses on the stability of these keto intermediates and emphasizes the major differences between indolizinones and pyrroloquinazolinones series. Noteworthy is also the report of some original structures isolated as by-products of some experiments.     

CoFe2O4自形成磁性液体场致结构化对磁化的影响

The Langevin paramagnetic theory can’t describe the relation between magnetization of ferrofluids and applied magnetic field. The structuralization of ferrofluids, which is considered the main influence factor of the magnetization, is regarded. The part of magnetization works is deposited when the structure is forming. This action influences the magnetization of ferrofluids directly or indirectly. On the base of the “compressing” model, the Langevin function that usually describes the magnetization of ferrofluid is modified, and a well-fitted curve is obtained. An equation of the relation between the equivalent volume fraction after being “compressed” and the intensity of magnetic field is discovered, which approximately describes the process of magnetization. The relation between the approximate initial susceptibility and the volume fraction can be obtained from modified formula.     

Highly regioselective Buchwald–Hartwig amination at C-2 of 2,4-dichloropyridine enabling a novel approach to 2,4-bisanilinopyridine (BAPyd) libraries

The highly regioselective Buchwald–Hartwig amination at C-2 of the cheap and readily accessible reagent, 2,4-dichloropyridine with a range of anilines and heterocyclic amines is described. This new methodology is robust and provides a facile access to 4-chloro-N-phenylpyridin-2-amines on 0.25 mol scale. These intermediates undergo a further Buchwald–Hartwig amination at higher temperature to enable rapid exploration of the chemical space at C-4 and to provide a library of 2,4-bisaminopyridines.     

KMnO4-mediated oxidative CN bond cleavage of tertiary amines: Synthesis of amides and sulfonamides

KMnO₄-mediated oxidative CN bond cleavage of tertiary amines producing secondary amine was introduced, which was trapped by electrophiles (acyl chloride and sulfonyl chloride) to form amides and sulfonamides. The reaction could take place at mild condition, tolerating a wide range of function groups and affording products in moderate to excellent yields.     

Chemistry of heterocyclic compounds at the A. E. Favorsky Irkutsk Institute of Chemistry, Siberian Branch of the Russian Academy of Sciences, over 50 years (Review)

The review contains a concise historical account and information on the most significant researches undertaken by the staff at the A. E. Favorsky Irkutsk Institute of Chemistry, Siberian Branch of the Russian Academy of Sciences on the Chemistry of Heterocyclic Compounds. Dedicated to Academician of the Russian Academy of Sciences B. A. Trofimov on his 70th jubilee. Translated from Khimiya Geterotsiklicheskikh Soedinenii, No. 10, pp. 1443–1502, October, 2008.     

Sulfur-directed metal-free and regiospecific methyl C(sp3)–H imidation of thioanisoles

Regiospecific and direct imidation of the methyl C(sp³)–H bond of thioanisoles is realized under mild and metal-free conditions with N-fluorobis(benzenesulfonyl)imide as an oxidant and nitrogen source. Proposed mechanism suggests that thionium ion intermediates and a Pummerer-type reaction are involved. The imidation has advantages such as high step-economy, excellent functionality tolerance, and regiospecificity, giving structurally diverse imidation products.     

Selective syntheses of 2H-1,3-oxazines and 1H-pyrrol-3(2H)-ones via temperature-dependent Rh(II)-carbenoid-mediated 2H-azirine-ring expansion

The Rh(II)-catalyzed reaction of 2-carbonyl-substituted 2H-azirines with ethyl 2-cyano-2-diazoacetate or 2-diazo-3,3,3-trifluoropropionate provides an easy access to 2H-1,3-oxazines and 1H-pyrrol-3(2H)-ones. These compounds can be selectively prepared from the same starting material using temperature as the only varied parameter. The 2-azabuta-1,3-diene intermediate, a common precursor for both heterocyclic products, isomerizes into 2H-1,3-oxazine under kinetic control, while 1H-pyrrol-3(2H)-one is the sole product of the reaction at elevated temperatures. According to DFT-calculations a one-atom oxazine ring contraction involving ring-opening to a 2-azabuta-1,3-diene intermediate, followed by a 1,5- and 1,2-prototropic shift leads to the consecutive formation of imidoylketene and azomethine ylide, which then further undergo cyclization to the pyrrole derivative.     

Synthesis of 1-benzyloxypyrazin-2(1H)-one derivatives

Different approaches for the synthesis of 1-benzyloxypyrazin-2(1H)-one derivatives from simple amino acids have been investigated. A library of 33 precursors for the preparation of N-hydroxy pyrazinones was obtained in moderate to good yields.