938 resultados para NADH Tetrazolium Reductase


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The aim of this study was to estimate the acute effects of low dose C-12(6+) ions or X-ray radiation on human immune function. The human peripheral blood lymphocytes (HPBL) of seven healthy donors were exposed to 0.05 Gy C-12(6+) ions or X-ray radiation and cell responses were measured at 24 h after exposure. The cytotoxic activities of HPBL were determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT); the percentages of T and NK cells subsets were detected by flow cytometry; mRNA expression of interleukin (IL)-2, tumor necrosis factor (TNF)-alpha and interferon (IFN)-gamma were examined by real time quantitative RT-PCR (qRT-PCR); and these cytokines protein levels in supematant of cultured cells were assayed by enzyme-linked immunosorbent assays (ELISA). The results showed that the cytotoxic activity of HPBL, mRNA expression of IL-2, IFN-gamma and TNF-alpha in HPBL and their protein levels in supernatant were significantly increased at 24 h after exposure to 0.05 Gy C-12(6+) ions radiation and the effects were stronger than observed for X-ray exposure. However, there was no significant change in the percentage of T and NK cells subsets of HPBL. These results suggested that 0.05 Gy high linear energy transfer (LET) C-12(6+) radiation was a more effective approach to host immune enhancement than that of low LET X-ray. We conclude that cytokines production might be used as sensitive indicators of acute response to LDL (C) 2009 COSPAR. Published by Elsevier Ltd. All rights reserved.

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实验目的:随着科技的发展,人类活动范围已经逐渐向外太空扩展,对于人类太空探索的最大威胁是太空中的各种粒子辐射。这些辐射包括太阳辐射(质子和电子)和银河辐射(质子占85%,氦离子占14%,重离子占1%)。众所周知,重离子与常规X和γ射线相比有较高的传能线密度(linear energy transfer, LET)和相对生物学效应(relative biological effectiveness, RBE),对机体组织和器官有较强的影响。放射治疗是肿瘤治疗的重要手段之一,由于肿瘤细胞的异质性,其对放、化疗的反应相差悬殊。本研究的目的是: 1评估辐射对健康机体产生的生物学风险; 2研究抗氧化剂氮乙酰半胱氨酸(NAC)对机体辐射损伤的保护作用 3不同肿瘤细胞辐射敏感性的差异。实验方法: 1 X射线或12C6+离子对小鼠进行不同剂量的全身辐射。NAC处理组小鼠在照射前1小时腹腔注射200mg/kg的NAC,对照组注射等体积的生理盐水。照射后不同时间点取样,利用流式细胞仪检测小鼠免疫细胞周期和凋亡情况,单细胞电泳检测淋巴细胞DNA损伤,MTT法(3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide)检测脾脏NK(natural killer,NK)细胞活性,微核法检测淋巴细胞染色体损伤情况,小鼠体内干扰素-γ(Interferon-γ,IFN-γ)由ELISA方法得到,小鼠血清中超氧化物岐化酶(Surperoxide dismutase SOD)由分光光度法测定,并观察胸腺和脾脏指数变化。 2 不同剂量X射线和12C6+离子辐射人肺腺癌细胞H1299和A549,用细胞克隆法检测照射后细胞存活曲线,流式细胞仪检测细胞周期和凋亡,Western-blot 检测A549 细胞P53蛋白表达。 结果: 1小鼠外周血淋巴细胞、胸腺细胞和脾脏淋巴细胞周期随着X射线照射剂量的增大而被阻滞在了G0/G1期,相同剂量的12C6+离子辐射时外周血淋巴细胞周期被阻滞在S期,分次连续X射线照射时,外周血淋巴细胞周期随着累积剂量的增加被阻滞在G2/M期;细胞凋亡比例随着照射剂量的增加而增加。小鼠血清中IFN-γ水平和脾脏中NK细胞活性在重离子照射剂量为0.05Gy时有显著增加,脾脏NK细胞活性随着照射剂量的增加而减弱。 2重离子照射后,小鼠淋巴细胞DNA和染色体的损伤随辐射剂量和照射后时间的延长而加剧。脾脏NK细胞活性在照射后各个时间点减弱,血清中IFN-γ水平和SOD酶活性随着重离子照射剂量的增加而降低。预防性给予NAC,12C6+离子辐射对淋巴细胞DNA和染色体所致损伤,胸腺细胞周期和凋亡,脾脏NK细胞活性,血清中IFN-γ的水平和SOD酶的活性的损伤与盐水组比较均有显著改善。 3 X射线照射对肺腺癌H1299细胞周期和凋亡率未产生明显影响,重离子照射后随着照射剂量的增加细胞周期被阻滞在G2/M期,细胞凋亡率也呈剂量依赖性;X射线和12C6+离子照射A549细胞后,细胞周期均被阻滞在G2/M期,凋亡率剂量依赖性增加。A549细胞P53蛋白的表达水平随着重离子照射剂量的增加而增加。结论: 1重离子辐射造成细胞DNA和染色体损伤随着照射剂量的增加和照射后时间的延长而增加,比X射线辐射损伤复杂和难以修复,产生这种现象的机理为辐射导致活性氧分子簇的产生,细胞因子和与细胞氧化反应有关的酶活性的变化,同时这种损伤对胸腺细胞周期、凋亡和胸腺、脾脏指数以及机体免疫系统都有影响;低剂量重离子辐射(0.05Gy)对小鼠机体的免疫力有刺激作用,机体免疫能力随着照射剂量增加和照射后时间的推移而减弱,不同的免疫器官对辐射的敏感性也不同; 2 200mg/kg 的NAC对辐射所致小鼠免疫系统损伤有很好的保护作用; 3 肺腺癌细胞H1299比同系A549具有较强的辐射敏感性,A549细胞凋亡的增加与P53蛋白表达水平升高有关

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根据ISO11268-1:1993方法,完成了土壤中苯并[a]芘对赤子爱胜蚓(Eisenia fetida)的人工污染模拟实验.并采用Clontech PCR-SelectTMcDNASubtration Kit构建了污染组和对照组蚯蚓之间的抑制消减文库.经测序和比对分析,在苯并[a]芘诱导上调文库中发现5个线粒体DNA(mtDNA)编码的基因,且在上调文库中具有较高的频率.它们分别为:细胞色素c氧化酶亚基Ⅰ、Ⅱ、Ⅲ,NADH氢脱氢酶亚基1,ATP合成酶亚基.研究表明,土壤中苯并[a]芘污染改变了蚯蚓线粒体基因组的表达水平,线粒体基因组表达异常是多环芳烃污染胁迫的重要分子生物标记物.

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The study on the response of a mutant and a wild type of Arabidopsis to 660 μl·L -1 CO 2 and ambient CO 2 showed that under elevated CO 2,the stomatal numbers of the mutant increased,while those of the wild type decreased. The chlorophyll content and NR (nitrate reductase) activity of the mutant increased,but those of the wild type had no obvious response. The mutant was not reproductively mature after the continuous exposure to increased CO 2 for five months. The results provided evidence of plant response to the changes of atmospheric CO 2 concentration,and the clues to related studies on other plants.

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In this work, the excel lent catalytic activity of highly ordered mesoporous carbons (OMCs) to the electrooxidation of nicotinamide adenine dinucleotide (NADH) and hydrogen peroxide (H2O2) was described for the construction of electrochemical alcohol dehydrogenase (ADH) and glucose oxidase (GOD)-based biosensors.

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In this paper, the unique properties of highly ordered mesoporous carbons modified glassy carbon electrode (OMCs/GE) are illustrated from comparison with carbon nanotubes modified glassy carbon electrode (CNTs/GE) for the electrochemical sensing applications.

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A fullerene/ionic-liquid composite was explored. Transmission Electron Microscopy (TEM) study showed that in the composite, C-60 mainly exists as nano-clusters, Raman spectrum proved that the composite formed only by physical Mix of C-60 and 1-Butyl-3-methyl-imidazolium hexafluorophosphate (BMIPF6), the combination did not change the chemical naturation of C-60. The electrochemical properties of the composite modified electrode, including the electrode reaction control function and the interfacial potential effect were studied.

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The size-controlled synthesis of monodispersed gold nanoparticles (AuNPs) stabilized by polyelectrolyte-functionalized ionic liquid (PFIL) is described. The resulting AuNPs' size, with a narrow distribution, can be tuned by the concentration of HAuCl4. Such PFIL-stabilized AuNPs (PFIL-AuNPs) showed a high stability in water at room temperature for at least one month; they were also quite stable in solutions of pH 7-13 and high concentration of NaCl.

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In this paper, the characterization and application of a chemically reduced graphene oxide modified glassy carbon (CR-GO/GC) electrode, a novel electrode system, for the preparation of electrochemical sensing and biosensing platform are proposed. Different kinds of important inorganic and organic electroactive compounds (i.e., probe molecule (potassium ferricyanide), free bases of DNA (guanine (G), adenine (A), thymine (T), and cytosine (C)), oxidase/dehydrogenase-related molecules (hydrogen peroxide (H2O2/beta-nicotinamide adenine dinucleotide (NADH)), neurotransmitters (dopamine (DA)), and other biological molecules (ascorbic acid (AA), uric acid (UA), and acetaminophen (APAP)) were employed to study their electrochemical responses at the CR-GO/GC electrode, which shows more favorable electron transfer kinetics than graphite modified glassy carbon (graphite/GC) and glassy carbon (GC) electrodes.

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This study demonstrates a novel compartment-less glucose/O-2 biofuel cell (BFC) based on highly ordered mesoporous carbons (OMCs) with three-dimensionally (3D) interconnected and ordered pore structures. OMCs are used as supports for both stably confining the electrocatalyst (i.e., meldola's blue, MDB) for NADH oxidation and the anodic biocatalyst (i.e., NAD(+)-dependent glucose dehydrogenase, GDH) for glucose oxidation, and for facilitating direct electrochemistry of the cathodic biocatalyst (i.e., laccase, LAC) for O-2 electroreduction. In 0.10 M pH 6.0 PBS containing 20 mM NAD(+) and 60 mM glucose under the air-saturated atmosphere, the open circuit voltage (0.82 V) and the maximum power output (38.7 mu W cm(-2) (at 0.54V)) of the assembled compartment-less OMCs-based BFC are both higher than those of carbon nanotubes (CNTs)-based BFC (0.75 V and 2.1 mu W cm(-2) (at 0.46 V)).

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Water-soluble supramolecular inclusion complexes of alpha-, beta-, and gamma-cyclodextrin-bicapped C-60 (CD/C-60) have been investigated for their photoinduced DNA cleavage activities, with the aim to assess the potential health risks of this class of compounds and to understand the effect of host cyclodextrins having different cavity dimensions. Factors such as incubation temperature, irradiation time, and concentration of NADH or CDs/C-60 supramolecular inclusion complexes have been examined. The results show that alpha-, beta-, and gamma-CDs/C-60 are all able to cleave double-stranded DNA under visible light irradiation in the presence of NADH. However, a difference in the photoinduced DNA cleavage efficiency is observed, where the cleavage efficiency increases in the order of alpha-, beta-, and gamma-CD/C-60. The difference is attributed to the different aggregation behavior of the inclusion complexes in aqueous solution, which is correlated to the cavity dimension of the host cyclodextrin molecules.

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Paclitaxel-loaded poly(ethylene glycol)-b-poly(L-lactide (LA)) (PEG-PLA) micelles were prepared by two methods. One is physical encapsulation of paclitaxel in micelles composed of a PEG-PLA block copolymer and the other is based on a PEG-PLA-paclitaxel conjugate, abbreviated as "conjugate micelles" Their physicochemical characteristics, e.g. critical micelle concentration (CMC), morphology, and micelle size distribution were then evaluated by means of fluorescence spectroscopy, scanning electron microscopy (SEM), and dynamic light scattering (DLS). The results show that the CMC of PEG-PLA-paclitaxel and PEG-PLA are 6.31 x 10(4) and 1.78 x 10(-3) g L-1, respectively. Both micelles assume a spherical shape with comparable diameters and have unimodal size distribution. Moreover, in vitro drug delivery behavior was studied by high performance liquid chromatography (HPLC). The antitumor activity of the paclitaxel-loaded micelles against human liver cancer H7402 cells was evaluated by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) method.

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A triblock poly(lactic acid)-b-poly(ethylene glycol)-b-poly(lactic acid) (PLA-PEG-PLA)/paclitaxel (PTX) conjugate was synthesized by the reaction of carboxyl-terminated copolymer PLA-PEG-PLA with PTX in the presence of dicyclohexylcarbodiimide and dimethylaminopyridine. Carboxyl-terminated copolymer PLA-PEG-PLA was prepared by the reaction of the hydroxyl end groups in copolymer PLA-PEG-PLA with succinic anhydride. Its structure was confirmed by NMR and gel permeation chromatography. The PLA-PEG-PLA/PTX conjugates could self-assemble into micelles in aqueous solutions with a low critical micelle concentration. Dynamic light scattering and environmental scanning electron microscopy analyses of the PLA-PEG-PLA/PTX micelles revealed their spherical structure and size of 220 nm. The antitumor activity of the conjugate against woman Hela cancer cells, evaluated by the 3-(4,5-dimethylthiazol2-yl)-2,5-diphenyl tetrazolium bromide method, showed that the conjugates had an antitumor activity similar to that of pure PTX. The obtained PLA-PEG-PLA/PTX conjugates are expected to be used in clinical practice.

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The homogeneous electrocatalytic reduction of 1,2-diiodoethane by anions of the supramolecular complex of (beta-CD)(2)/C-60 in DMF solution is reported. The results show that the trianion of (beta-CD)(2)/C-60 exhibits electrocatalytic behavior towards the reduction of 1,2-diiodoethane, whereas the diani on is unable to reduce the diiodoethane. The second-order catalytic rate constant in DMF solution was determined to be 3.1 x 10(5) M-1 s(-1) by analysis of voltammetric responses under pseudo-first-order conditions with respect to (beta-CD)(2)/C-60. The results suggest that the host beta-cyclodextrin molecules have little effect on the electrocatalytic ability of the encapsulated C-60 toward organic halides.

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The report described a method of more stably dispersing oxidized carbon nanotubes (CNTs) by forming complex with polycation and the layer-by-layer self-assembly behavior of the complex with polyanion was studied. The properties of the self-assembled multilayer film containing carbon nanotubes were studied. Cyclic voltammetry, UV-vis-NIR spectroscopy, electrochemical impedance spectroscopy and scanning electron microscopy were used for characterization of film assembly. UV-vis-NIR spectroscopy and cyclic voltammetry study indicated the uniform growth of the film. Electrochemical impedance spectroscopy results showed that incorporating of carbon nanotubes in the polyelectrolyte multilayers; decreased in the electron-transfer resistance R, indicating more favorable electrochemical reaction interface. The electrocatalytic property of the multilayer modified electrode to NADH was investigated mainly with different numbers of the bilayers; and the results showed that along with the increase of the assembled bilayers the overpotential of NADH oxidation decreased. The detection lit-nit Could reach 6 mu M at a detection potential of 0.4 V.