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981.
目的:探讨一氧化氮供体药物硝酸甘油对对裸鼠胰腺癌皮下成瘤的抑制作用及对瘤体组织中低氧诱导因子1α(hypoxia induced factor 1α,HIF-1α),P糖蛋白(P-glucoprotein,P-gP)和谷胱甘肽S转移酶-π(Glutathione S-transferase pi,GST-π)的表达影响.方法:40只6-8周的18-22g的BALB/c裸鼠双侧腹股沟皮下注射胰腺癌PANC-1细胞成瘤,瘤体超过150mm3后随机分组,干预组用硝酸甘油帖剂干预,对照组贴安慰剂,并分别绘制生长曲线.采用免疫组织化学方法检测成瘤裸鼠中瘤体组织中HIF-1α、P-gP和GST-π的表达.结果:32例裸鼠成瘤,成瘤率80%,干预28天后,干预组和对照组的瘤体大小分别为1550.4±148.6mm3和750.6±71.2mm3.16只对照组瘤体组织中HIF-1α阳性表达12例(75%),P-gp阳性表达10例(62.5%),GsT-π阳性表达8例(50%).在16例干预组裸鼠瘤体组织中HIF-1α、P-gP、GST-π阳性表达分别是11例(68.8%),8例(50%),7例(43.8%).结论:特定剂量硝酸甘油对裸鼠皮下成瘤有明显抑制作用,且对HIF-1α及P-gp、GsT-π的表达也有一定的抑制作用. 相似文献
982.
1998年,Thomson等成功分离出了人类胚胎干细胞(embryonic stem cell,ES)系为研究再生医学和人类早期发育开辟了一个新领域.通过转染人类ES细胞进行遗传操作(genetic manipulation)可以帮助我们更好地理解人类Es细胞的生物学特性,同时为研究人类ES细胞在细胞及基因治疗中潜在的治疗用途提供了基础.本文综述了人类ES细胞外源基因转染方法及其应用的进展. 相似文献
983.
984.
Lijuan Zhang Zhaoqin Zhu Huaiqi Jing Jingyun Zhang Yanwen Xiong Meiying Yan Shouyi Gao Long-Fei Wu Jianguo Xu Biao Kan 《BMC microbiology》2009,9(1):114-13
Background
The Twin-arginine translocation (Tat) system serves to translocate folded proteins, including periplasmic enzymes that bind redox cofactors in bacteria. The Tat system is also a determinant of virulence in some pathogenic bacteria, related to pleiotropic effects including growth, motility, and the secretion of some virulent factors. The contribution of the Tat pathway to Vibrio cholerae has not been explored. Here we investigated the functionality of the Tat system in V. cholerae, the etiologic agent of cholera. 相似文献985.
Background
The phylogeny of Cetacea (whales) is not fully resolved with substantial support. The ambiguous and conflicting results of multiple phylogenetic studies may be the result of the use of too little data, phylogenetic methods that do not adequately capture the complex nature of DNA evolution, or both. In addition, there is also evidence that the generic taxonomy of Delphinidae (dolphins) underestimates its diversity. To remedy these problems, we sequenced the complete mitochondrial genomes of seven dolphins and analyzed these data with partitioned Bayesian analyses. Moreover, we incorporate a newly-developed "relaxed" molecular clock to model heterogenous rates of evolution among cetacean lineages. 相似文献986.
Xia Xiong Sha Huang Hai Zhang Jianjun Li Jianying Shen Jixian Xiong Yong Lin Liping Jiang Xianchun Wang Sonping Liang 《Proteome science》2009,7(1):41-10
Background
Dorsal root ganglion (DRG) neurons are primary sensory neurons that conduct neuronal impulses related to pain, touch and temperature senses. Plasma membrane (PM) of DRG cells plays important roles in their functions. PM proteins are main performers of the functions. However, mainly due to the very low amount of DRG that leads to the difficulties in PM sample collection, few proteomic analyses on the PM have been reported and it is a subject that demands further investigation. 相似文献987.
Effects of elevated CO2 concentration, supplemental irrigation and nitrogenous fertilizer application on rain-fed spring wheat yield 下载免费PDF全文
It is expected that the CO2 concentration of the Earth’s atmosphere will reach 600–1000 ppm by the end of the 21st century. Therefore, in this study, we evaluated the effects of elevated CO2 concentrations on the development of rain-fed spring wheat in an attempt to identify a practical pathway to increase crop production. To accomplish this, a field experiment was conducted at Guyuan Experimental Station in a semiarid region of China during 2005–2007. During this experiment, the CO2 concentration was increased to 40.0 ppm and supplemental irrigation and nitrogenous fertilizer (N fertilizer) were applied. The experimental results showed that the elevated CO2 concentration significantly improved the thousand-grain weight and the grain number per spike. Furthermore, supplemental irrigation and N fertilizer application during the elongation and booting stage of rain-fed spring wheat in conjunction with an elevated CO2 concentration improved the water use efficiency (WUE), nitrogen use efficiency (NUE), thousand-grain weight, and the yield by 14.6%, 39.6%, 9.3%, and 14.7%, respectively, when compared to groups subjected to the same treatment but not grown under elevated CO2 concentrations. Furthermore, the spring wheat yield was improved by 81.8% in response to an elevated CO2 concentration, 60 mm of supplemental irrigation and applied N fertilizer (37.5 g m?2 NH4NO3). However, the presence of an elevated CO2 concentration without supplemental irrigation and N fertilizer only resulted in an increase in the wheat yield of 7.8%. Consequently, the combination of elevated CO2 concentration, supplemental irrigation and N fertilizer application played an important role in the improvement of WUE, NUE, thousand-grain weight, and grain yield of rain-fed spring wheat in this region. 相似文献
988.
Nian Xiong Jinsha Huang Zhentao Zhang Zhaowen Zhang Jing Xiong Xingyuan Liu Min Jia Fang Wang Chunnuan Chen Xuebing Cao Zhihou Liang Shenggang Sun Zhicheng Lin Tao Wang 《PloS one》2009,4(11)
A clinically-related animal model of Parkinson''s disease (PD) may enable the elucidation of the etiology of the disease and assist the development of medications. However, none of the current neurotoxin-based models recapitulates the main clinical features of the disease or the pathological hallmarks, such as dopamine (DA) neuron specificity of degeneration and Lewy body formation, which limits the use of these models in PD research. To overcome these limitations, we developed a rat model by stereotaxically (ST) infusing small doses of the mitochondrial complex-I inhibitor, rotenone, into two brain sites: the right ventral tegmental area and the substantia nigra. Four weeks after ST rotenone administration, tyrosine hydroxylase (TH) immunoreactivity in the infusion side decreased by 43.7%, in contrast to a 75.8% decrease observed in rats treated systemically with rotenone (SYS). The rotenone infusion also reduced the DA content, the glutathione and superoxide dismutase activities, and induced alpha-synuclein expression, when compared to the contralateral side. This ST model displays neither peripheral toxicity or mortality and has a high success rate. This rotenone-based ST model thus recapitulates the slow and specific loss of DA neurons and better mimics the clinical features of idiopathic PD, representing a reliable and more clinically-related model for PD research. 相似文献
989.
Changhui Deng Xinghua Xiong Andrew N. Krutchinsky 《Molecular & cellular proteomics : MCP》2009,8(6):1413-1423
We have developed and applied a method unifying fluorescence microscopy and mass spectrometry for studying spatial and temporal properties of proteins and protein complexes in yeast cells. To combine the techniques, first we produced a variety of DNA constructs that can be used for genomic tagging of proteins with modular fluorescent and affinity tags. The modular tag consists of one of the multiple versions of monomeric fluorescent proteins fused to a variety of small affinity epitopes. After this step we tested the constructs by tagging two yeast proteins, Pil1 and Lsp1, the core components of eisosomes, the large protein complexes involved in endocytosis in Saccharomyces cerevisiae, with a variety of fluorescent and affinity probes. Among the modular tags produced we found several combinations that were optimal for determining subcellular localization and for purifying the tagged proteins and protein complexes for the detailed analysis by mass spectrometry. And finally, we applied the designed method for finding the new protein components of eisosomes and for gaining new insights into molecular mechanisms regulating eisosome assembly and disassembly by reversible phosphorylation and dephosphorylation. Our results indicate that this approach combining fluorescence microscopy and mass spectrometry into a single method provides a unique perspective into molecular mechanisms regulating composition and dynamic properties of the protein complexes in living cells.Fluorescent proteins have become invaluable probes for studying molecular processes in living cells with light microscopy techniques (1–3). Proteins, organelles, and entire cells can be selectively visualized using a variety of fluorescent proteins fused to the proteins of interest (1–6). Combined with genetics and molecular biology techniques fluorescence microscopy provides an efficient tool for observing molecular phenotypes useful for dissecting the pathways of cell cycle progression and cell response to internal and external signals (7). However, understanding the mechanism controlling the properties of proteins in cells can be a challenging task, frequently requiring a comprehensive characterization of the proteins at the molecular level.The proteins tagged with green fluorescent protein (GFP)1 can be also purified using GFP antibodies. Cheeseman and Desai (8) and Cristea et al. (9) have enriched GFP-tagged proteins and protein complexes for further detailed analysis by MS. The MS-based methods for protein analysis are fast, sensitive, and able to identify both proteins in complex protein mixtures and residues bearing post-translational modifications (10, 11). Thus, the addition of affinity purification and mass spectrometry steps enabled the researchers to study protein interactions and the post-translational modifications in the context of the protein subcellular localization. Juxtaposition of the protein localization, composition of the protein complexes, and post-translational modifications frequently yield a unique perspective of the cellular processes and the molecular mechanisms of their regulation (12, 13).Using fluorescent proteins also as affinity probes can be problematic in several instances. First of all, the good quality antibodies against the rapidly increasing number of fluorescent proteins (3, 6) are not yet readily available. Furthermore raising antibodies specifically recognizing fluorescent proteins originating from the same organism but fluorescing a different color can be difficult or even impossible because such proteins frequently differ by mutations of only a few amino acids (1–6). Thus, we seek an alternative approach to the design of tags suitable for subcellular localization and purification of proteins and protein complexes that is 1) independent of the availability of antibody to a specific form of a fluorescent protein, 2) suitable for multiplexing, i.e. simultaneous observation of subcellular localization of several proteins and affinity purification of the proteins and stably associated protein complexes, and 3) flexible and easy to modify to incorporate better versions of fluorescent proteins and affinity tags after they are discovered.One possible solution that satisfies the stated requirements is to use a modular tag containing a version of a fluorescent protein fused to an affinity epitope. In this case we can decouple requirements for both modules and optimize the performance of each one independently for fluorescence microscopy and affinity purification experiments. To our knowledge, this possibility was first realized by Thorn and co-worker (14) who have fused 3HA (three repeats of YPYDVPDYA epitope from hemagglutinin protein) and 13MYC (13 repeats of EQKLISEEDL epitope, corresponding to a stretch of the C-terminal amino acids of the human c-MYC protein) tags to several variants of fluorescent proteins. The authors have argued that the fusion of the fluorescent proteins to the affinity epitopes may enable fluorescence and immunochemical analysis but did not test this idea. Cheeseman and Desai (8) fused the S-peptide and hexahistidine epitopes to the GFP protein to enable additional tandem purification steps. Su and co-workers (15) also fused a hexahistidine tag (His6) to GFP to purify recombinantly produced proteins. Although hexahistidine tag performs well for isolation of overexpressed recombinant proteins, it works poorly for affinity purification of low abundance, endogenously expressed proteins (16). A double affinity tag containing a single MYC epitope and hexahistidine was also used to purify recombinantly produced fluorescent proteins (6).Here we describe the design and implementation of the modular fluorescent and affinity tags. These tags contain a variety of fluorescent proteins, which can be used exclusively for obtaining subcellular visualization, and several small epitope tags that can be utilized to perform two-step affinity purification. To test the performance of the constructs produced, we tagged two yeast proteins, Pil1 and Lsp1, the core components of eisosomes, with a variety of modular tags.Eisosomes are large heterodimeric protein complexes recently discovered in Saccharomyces cerevisiae (17). There are ∼50–100 eisosomes in each mature yeast cell distributed uniformly in a characteristic dotted pattern at the cell surface periphery. Each eisosome contains ∼2000–5000 copies of Pil1 and Lsp1. It was shown that eisosomes serve as portals of endocytosis in yeast. The function of eisosomes is regulated by reversible phosphorylation (18, 19).Among the constructs tested, we found several combinations of fluorescent protein and affinity tags that were optimal for determining subcellular localization and purification of the proteins and protein complexes. We applied these tags to further investigate eisosomes and found several new protein components of the complexes and obtained new insights into molecular mechanisms regulating eisosome integrity by reversible phosphorylation and dephosphorylation. Our results indicate that an approach combining fluorescence microscopy and mass spectrometry into a single method provides a unique perspective into molecular mechanisms regulating composition and dynamic properties of the protein complexes in living cells. 相似文献
990.
Tomomi Kiyota Masaru Yamamoto Huangui Xiong Mary P. Lambert William L. Klein Howard E. Gendelman Richard M. Ransohoff Tsuneya Ikezu 《PloS one》2009,4(7)