恒河猴TRIM5α的组织分布及不同刺激促进PBMC中TRIM5α转录水平的上调

TRIM5α(tripartite motif protein 5-alpha)蛋白是恒河猴体内一种非常重要的限制因子,能抑制人免疫缺陷病毒(HIV-1,human immunodeficiency virus type 1)、马感染性贫血病毒(EIAV, equine infectious anemia virus)和猫免疫缺陷病毒(FIV, feline immunodeficiencyvirus)等逆转录病毒的复制.恒河猴TRIM5α的组织分布以及在受到外界刺激时TRIM5α mRNA表达量的变化研究还未见报道.本研究从中国恒河猴的各组织中提取总RNA,以β-actin基因作为内参照,通过半定量RT-PCR检测各组织中TRIMSα mRNA的表达.选择HIV-GFP-VSVG假病毒感染外周血单核细胞(peripheral blood mononuclear cell,PBMC),非特异性刺激剂--佛波脂(Phorbol myfismte acetate,PMA)+离子霉素(ionomycin,Ion)及CD28抗体+CD49d抗体分别共刺激恒河猴PBMC,研究不同刺激对恒河猴TRIM5α mRNA表达水平的影响.结果表明:TRIM5α mRNA表达于所研究的恒河猴21种组织中,免疫系统和泌尿生殖系统组织中表达量最高,而神经系统组织,如大脑、脊髓中表达较少,其他组织中未见明显的表达差异;HIV-GFP-VSVG感染和用PMA+Ion、CD28抗体+CD49d抗体分别共刺激PBMC能促进PBMC中TRIM5α mRNA的转录水平的上调.

蛙虹彩病毒cop基因的克隆表达及亚细胞定位

从蛙虹彩病毒(Rana gryliovirus,RGV)基因组中克隆了含凋亡相关结构域的新基因-cop(Caspaserecruit ment domain only protein,COP)基因的全部编码区,成功构建了重组表达载体,进行了原核表达,并在鲤鱼上皮瘤细胞(Epithelioma papulosumcyprini,EPC)中进行了亚细胞定位.序列分析表明,RGVcop基因全长288 bp,编码一个长为95 aa,分子量为10.4×103的推定蛋白.二级结构预测表明其含有5个α螺旋.同源性比对分

恒河猴TRIM5α 的组织分布及TRIMCyp 限制HIV-1 病毒颗粒产生的初步研究

三重基序蛋白TRIM5α(Tripartite motif protein 5 alpha)是哺乳动物细胞中一种重要的限制因子，广泛分布于各种哺乳动物细胞中。人类TRIM5α mRNA 广泛表达于人类各个组织中，并且I 型干扰素IFN-α/β/γ 均能与TRIM5α 基因启动子的ISRE 元件结合，上调TRIM5α mRNA 的表达。恒河猴(Macaca mulatta)TRIM5α 是恒河猴体内重要的限制因子。目前对恒河猴尤其是中国恒河猴TRIM5α 的组织分布以及在受到外界刺激时TRIM5α mRNA 表达量的变化研究还未见报道。本论文通过从中国恒河猴各组织中提取总RNA，以β-actin 基因作为内参照，通过逆转录PCR 检测各组织中TRIM5α mRNA 的表达。我们选择用HIV-GFP-VSVG 感染、用佛波脂(Phorbol myfismte acetate, PMA)+离子霉素(Ionomycin, Ion)，CD28 抗体+CD49d 抗体分别共刺激恒河猴PBMC，研究不同刺激对中国恒河猴TRIM5α mRNA 表达量的影响。研究发现：TRIM5α mRNA 广泛表达于恒河猴各组织中，在免疫系统和泌尿生殖系统各组织，如腹淋巴结、睾丸和附睾中表达量最高，而在神经系统各组织如大脑、脊髓中表达量比较少，在其他各组织中未见明显的表达差异。此外HIV-GFP-VSVG 感染、PMA+ Ion 与CD28 抗体+CD49d 抗体分别共刺激PBMC 均能促进PBMC TRIM5α mRNA 表达量的上调。 TRIM5α 作为恒河猴体内的最主要的限制HIV-1 感染的限制因子，除了可能通过促进HIV-1 的脱壳和阻止整合前复合物PIC(pre-integration complex)入核，恒河猴TRIM5α 还能限制HIV-1 病毒颗粒的产生。在这个过程中B30.2 结构域是非必需的，而B-box2 和Coiled-Coil 结构域起着决定性的作用。因为鹰猴(Aotes trivirgatus)TRIMCyp(omTRIMCyp) 蛋白和北平顶猴(Macaca leouina) TRIMCyp(npmTRIMCyp）蛋白的B-box2 和Coiled-Coil 结构域与恒河猴TRIM5α 的B-box2 和Coiled-Coil 具有很高的同源性，我们希望了解鹰猴TRIMCyp 蛋白和北平顶猴TRIMCyp 蛋白对HIV-1 病毒颗粒的产生是否有限制作用。本论文主要通过将质粒pNL4.3 分别与质粒pLPCX 、pLPCX-npmTRIMCyp-HA 、 pLPCX-omTRIMCyp-HA和pLPCX-rhTRIM5α-HA共转染293T细胞，通过western blot 检测细胞内Gag 蛋白和TRIM5 蛋白的表达情况，研究omTRIMCyp 蛋白和 npmTRIMCyp 蛋白对HIV-1 病毒颗粒产生的限制作用。结果表明：北平顶猴 TRIMCyp 蛋白、鹰猴TRIMCyp 蛋白都能不同程度的促进HIV-1 病毒Gag 蛋白的降解。

Evaluation of ferricyanide effects on microorganisms with multi-methods

In this study, we report the effects of ferricyanide on organisms based on the changes in physiological state and morphology of Escherichia coli (E coli) DH 5 alpha after being pretreated by ferricyanide. The impact on bacterial cell growth and viable rate of exposure to different concentrations of ferricyanide was determined, and the morphology change of E. coli was studied by atomic force microscopy (AFM). Finally, recovery test was used to evaluate the recovery ability of injured cells. The results showed that the effects on growth and morphology of E. coli were negligible when the concentration of ferricyanide was below 25.0 mM. While the results showed 50.8% inhibition of growth in the presence of 50.0 mM ferricyanide for 3 h, 89.6% viability was detected by flow cytometry (FCM) assay. AFM images proved that compact patches appeared on the bacterial surface and protected the bacterial viability. Furthermore, the results revealed that deterioration of bacterial surface closely related to the incubation time from 0.5 to 3 h at 100.0 mM ferricyanide. In the recovery test, microbial cell population and dissolved oxygen individually decreased 36.7% and 28.3% with 25.0 mM ferricyanide.

Amorphane sesquiterpenes from a marine Streptomyces sp.

The chemical investigation of the crude extract of the marine-derived Streptomyces sp. M491 yielded three new sesquiterpenes, namely, 10 alpha,11-dihydroxyamorph-4-ene (4), 10 alpha,15-dihydroxyamorph-4-en-3-one (6), and 5 alpha,10 alpha,11-trihydroxyamorphan-3-one (7). In addition, the known compounds 10 alpha-hydroxyamorph-4-en-3-one (2), o-hydroxyacetanilide, genistein, prunetin, and indole-3-carbaldehyde and the macrolide antibiotic chalcomycin A were identified. The structures were determined on the basis of spectroscopic analysis, especially 1D and 2D NMR data. This is the first report of these sesquiterpenes from bacteria.

Chemical Constituents of a Marine-Derived Endophytic Fungus Penicillium commune G2M

Cultivation of the endophytic fungus Penicillium commune, which was isolated from the semi-mangrove plant Hibiscus tiliaceus, afforded one new compound 1-O-(2,4-dihydroxy-6-methylbenzoyl)-glycerol (1) along with thirteen known products, including 1-O-acetylglycerol (2), N-acetyltryptophan (3), 3-indolylacetic acid methyl ester (4), 1-(2,4-dihydroxy-3,5-dimethylphenyl)ethanone (5), 2-(2,5-dihydroxyphenyl)acetic acid (6), (4R,5S)-5-hydroxyhexan-4-olide (7), thymidine (8), uracil (9), thymine (10), ergosterol (11), beta-sitosterol (12), beta-daucosterol (13), and ergosta-7,22-dien-3 beta,5 alpha,6 beta-triol (14). The structures of these compounds were established by detailed NMR spectroscopic analysis, as well as by comparison with literature data or with authentic samples.

Ergosterimide, a new natural Diels-Alder adduct of a steroid and maleimide in the fungus Aspergillus niger

Ergosterimide (1), a natural Diels-Alder adduct of ergosteroid and maleimide, was characterized from the culture extract of Aspergillus niger EN-13, an endophytic fungus isolated from the marine brown alga Colpomenia sinuosa. In addition, four known steroids including (22E,24R)-ergosta-5,7,22-trien-3 beta-ol (2), (22E,24R)-ergosta-4,6,8(14),22-tetraen-3one (3), (22E,24R)-5 alpha,8 alpha-epidioxyergosta-6,22-dien-3 beta-ol (4), and (22E,24R)-ergosta-7,22dien-3 beta,5 alpha,6 beta-triol. (5) were also isolated and identified. The structures of these compounds were elucidated by extensive analysis of 1D and 2D NMR and IR spectra and MS data. The plausible biosynthetic pathway of 1 was also discussed. To the best of our knowledge, 1 is the first natural Diels-Alder adduct of steroid and maleimide reported so far. (C) 2007 Elsevier Inc. All rights reserved.

Two new diterpenes from Euphorbia kansuensis

Two new lathyrane diterpenes, 3 beta,5 alpha-dihydroxy-15 beta-cinnamonyloxy-14-oxoladiyra-6Z, 12E- diene (1) and 3 beta,5 alpha,20-trihydroxy-15 beta-cinnamoyloxy-14-oxolathyra-6Z, 12E-diene (2), were isolated from the roots of Euphorbia kansuensis. Their structures were determined by spectroscopic methods. (C) 2008 Elsevier B.V. All rights reserved.

A new guaiane diterpenoid from Euphorbia wallichi

A new guaiane-type diterpenoid, (1 alpha, 5 alpha, 7 alpha)-3,10(18), 11-dictytriene-19-acid, was obtained from the roots of Euphorbia wallichii. This is the first isolation of guaiane diterpene from this genus of Euphorbia. The structure was elucidated by spectral methods. And the compound was tested for the cytotoxicities on the cancer cell line P-388 and A-549 in vitro.

Synthesis of 2-benzylthio-5-phenyl-3,4-disubstituted thiophenes by intramolecular condensation of alpha-oxo ketene dibenzylthioacetals

A facile route to 2-benzylthio-5-phenyl-3,4-disubstituted thiophenes was described. Catalyzed by sodium hydroxide, the title compounds were synthesized in moderate to good yields simply from the intramolecular aldol type condensation of alpha-oxo ketene dibenzylthioacetals. The chemical selectivity for this annulation reaction was studied and discussed.

SYNTHESIS AND MOLECULAR-STRUCTURE OF THE 5-COORDINATE ZIRCONIUM COMPLEX (CH3C5H4)2ZRCL(ALPHA-C10H7COO)

Bis(methylcyclopentadienyl)chloro(alpha-naphthoyloxy)zirconium, (CH3C5H4)2Zr Cl(alpha-C10H7COO), has been synthesized by the reaction between (MeCp)2ZrCl2 and equimolar sodium alpha-naphthoate in toluene at room temperature. The molecular structure of the title complex was determined by X-ray diffraction. The molecules have a five-coordinate bent metallocene structure in which the alpha-naphthoxyligand is bidentate [Zr-O, 2.317(2), 2.260(2) angstrom]. The remaining distances are [Zr-Cl, 2.521(1) angstrom; Zr-C, 480-2, 565 angstrom; Zr-cent-CH3C5H4 2.217 and 2.230 angstrom].

Edge-defined, film-fed growth (EFG) technique for the preparation of alpha-Al2O3 : C crystal with highly sensitive thermoluminescence response

In this work, alpha-Al2O3:C, a highly sensitive thermoluminescence dosimetry crystal, was grown by the EFG method in which a graphite heating unit and shield acted as the carbon source during the growth process. The optical, luminescent properties and dosimetric characteristics of the crystal were investigated. The as-grown crystal shows a single glow peak at 536 K, which is associated with Cr3+ ions. After annealing in H-2 at 1673 K for 80 h, the crystal shows a single glow peak at 460 K and a blue emission band at 415 nm. The thermoluminescent response of the annealed crystal shows linear-sublinear-saturation characteristics in the dose range from 5 x 10(-6) to 100 Gy.

Growth of highly sensitive thermoluminescent crystal alpha-Al2O3 : C by the temperature gradient technique

In this work, an alpha-Al2O3:C crystal with highly sensitive thermoluminescence was directly grown by the temperature gradient technique (TGT) using Al2O3 and graphite powders as raw materials. The optical and luminescent properties and the dosimetric characteristics of the crystal were investigated. An as-grown alpha-Al2O3:C crystal shows a single glow peak at 462 K and a blue emission peak at 415 nm. The thermoluminescence (TL) response of the crystal shows a linear-sublinear-saturation characteristic. In the dose range from 5 x 10(-6) to 10Gy, the alpha-Al2O3:C crystal shows excellent linearity, and saturation was observed at about 30Gy. The sensitivity of the crystal decreases as the heating rate increases. (c) 2008 Elsevier B.V. All rights reserved.

Growth of alpha-Al2O3:C crystal with highly sensitive optically stimulated luminescence

In this work. an alpha-Al2O3:C crystal was directly grown by the temperature gradient technique (TGT) using Al2O3 and graphite powders as the raw materials. The optical, optically stimulated luminescence (OSL) properties and dosimetric characteristics of as-grown crystal were investigated. As-grown alpha-Al2O3:C crystal shows strong absorption band at 205, 230 and 256 nm. Three-dimensional thermoluminescence (TL) emission spectrum of the crystal shows a single emission peak at similar to 415 nm. The OSL decay curve can be fitted to two exponentials, the faster component and the slower component. The OSL response of the crystal shows a linear-sublinear-saturation characteristic. As-grown alpha-Al2O3:C crystal shows excellent linearity in the dose range from 5 x 10(-6) to 50 Gy. For doses higher than the saturation dose (100 Gy). the OSL sensitivity decreases as the dose increases. Crown Copyright (C) 2008 Published by Elsevier B.V. All rights reserved.

Effects of process on microstructure and properties of translucent Dy-alpha-SiAlON sintered at lower temperatures

Hot pressing (HP) at higher sintering temperature has been a traditional and prevalent technique for the fabrication of alpha-SiAlON. In order to prepare translucent SiAlON more easily, LiF was used as a non-oxide sintering additive to lower the sintering temperature to <= 1650 degrees C. As a result, all of the samples possessed a good hardness and fracture toughness. At the same time, the lower temperature sintered samples showed a higher optical transmittance in the range of 2.5-5.5 mu m wavelength (0.5 mm in thickness). The maximum infrared transmission reached 68% at a wavelength of 3.3 mu m. The present work shows that the sintering process has a strong effect on microstructure and property of alpha-SiAlON. To be exact, a lower sintering temperature and longer holding time can produce some fully-developed microstrcture, which is beneficial for the optical transmittance. (C) 2008 The Ceramic Society of Japan. All rights reserved.