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骨巨细胞瘤组织S100A8和S100A9表达及与肿瘤影像学表现的相关性分析

浙江大学学报（医学版） 2015年第3期44卷 329-334页

作者：廖金生丁晓毅许顺良浙江大学医学院附属第一医院放射科浙江杭州310003 上海交通大学医学院附属瑞金医院放射科上海200025

目的：检测骨巨细胞瘤患者肿瘤组织中S100A8、S100A9mRNA及蛋白的表达水平，探讨其与骨巨细胞瘤的影像学及生物学行为之间的关系。方法：收集2009年1月至2012年6月在上海交通大学医学院附属瑞金医院诊治的43例骨巨细胞瘤患者的临床资料... 详细信息

目的：检测骨巨细胞瘤患者肿瘤组织中S100A8、S100A9mRNA及蛋白的表达水平，探讨其与骨巨细胞瘤的影像学及生物学行为之间的关系。方法：收集2009年1月至2012年6月在上海交通大学医学院附属瑞金医院诊治的43例骨巨细胞瘤患者的临床资料，并采集骨巨细胞瘤新鲜组织和6例正常成人股骨的骨髓组织，运用半定量逆转录PCR及蛋白质印迹法检测肿瘤组织中S100 A8、S100 A9 mRNA及其蛋白表达水平，并分析与不同肿瘤影像学表现如CT检查显示的骨质破坏程度，MRI检查显示的实性成分比例及周边水肿程度等关系。结果：43份骨巨细胞瘤组织中有40份表达S100 A8、S100 A9 mRNA及其蛋白，与正常骨髓组织比较，骨巨细胞瘤组织S100 A8、S100 A9 mRNA及其蛋白表达增加，差异均有统计学意义（均P＜0．05）。影像学不同实质成分比例的骨巨细胞瘤患者S100 A8蛋白表达差异具有统计学意义（ P ＜0．05）；不同骨质破坏程度的骨巨细胞瘤患者S100 A9蛋白表达差异也有统计学意义（ P ＜0．05）。结论：骨巨细胞瘤组织中S100 A8和S100 A9表达增加，并与骨巨细胞瘤骨质破坏程度及实性成分比例、水肿程度具有一定相关性，S100 A8、S100 A9可能参与骨巨细胞瘤的发生发展。

关键词：巨细胞瘤骨/病理学巨细胞瘤,骨/放射摄影术 RNA 信使/遗传学 S100蛋白质类/生物合成体层摄影术 X线计算机磁共振成像

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Effects of hypoxic preconditioning on the changes of expression of neuroglobin mRNA and labeled positive cells following cerebral ischemia in gerbils

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Neural Regeneration Research 2006年第5期1卷 397-400页

作者： Yong Zhang Yanqun Chang Zhenfang Liu Qingxi Fu Xiao Zhang Department of Neurology Shandong Provincial HospitalShandong University Jinan250021 Shandong ProvinceChina Depayment of Anesthesiology Shandong Provincial HospitalShandong University Jinan250021 Shandong ProvinceChina

BACKGROUND： Neuroglobin （NGB）, as newly discovered the third member of the globin family binding oxygen, mainly exists in brain of human beings and vertebrates, and it is closely correlated with the oxygen supply in brain. OBJECTIVE： To observe the changes of the expression of neuroglobin and number of positive cells labeled immunohistochemically following cerebral ischemia in gerbils after hypoxic preconditioning. DESIGN ： A complete randomized grouping design and controlled experiment. SETTING ： Department of Neurology, Department of Anesthesia, Shandong Provincial Hospital, Shandong University. MATERIALS ： Sixty-six adult male Mongolian gerbils of clean degree, about 50-65 g, at an average of 57.5 g were provided by the Experimental Animal Center of Capital Medical University [certificate number of animal quality：SCXK（Beijing）2000-0012]. TRNzil （Tianwei Shidai Company, Beijing）, polymerase chain reaction （PCR） primers （synthetized by Invitrogen Company, Shanghai）; reverse transcription-PCR （RT-PCR） one-step kit （Toyobo Company）; PCR instrument （GeneAmp PCR System 240）; mice brain NGB monoclonal antibody （Academy of Military Medical Sciences）; DAB （Zhongshan Company, Beijing）. METHEDS ： The study was completed from December 2004 to June 2005 in Shandong Provincial Hospital. ① The 66 gerbils were randomly divided into sham-operated group （n =6）, cerebral ischemia group （n =30） and hypoxic preconditioning group （n =30）. The gerbils in the hypoxic preconditioning group were put in the environment which contained 02 （0.08 in volume fraction） and N2 （0.92 in volume fraction） at temperature of 25 ℃ for 2 hours. After 5 hours, the gerbils in the hypoxic preconditioning group and cerebral ischemia group were anesthetized, then bilateral common carotid arteries were ligated. In the sham-operated group, bilateral common carotid arteries were only isolated without ligation and hypoxic preconditioning. ② At 1, 5, 10, 30 and 60 minutes after cerebral ischemia, the exp

关键词： RNA 信使/遗传学疾病模型动物脑缺血/代谢脑缺血/病理学缺血预处理沙鼠亚科一氧化氮/代谢成年人动物男(雄)性人类小鼠

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Pep-1-vMIP-II 对小鼠抗HIV-I基因表达的作用

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基础医学与临床 2013年第12期33卷 1538-1543页

作者：高媛南玉龙杨磊谭晓华尹洪萍汪浩石河子大学/新疆地方与民族高发病教育部重点实验室石河子市人民医院普外科新疆石河子832002 杭州师范大学健康管理学院杭州师范大学医学实验中心浙江杭州310036

目的:探讨融合蛋白Pep-1-vMIP-II在影响宿主ARGs基因表达抵抗HIV-I感染中的作用。方法:小鼠背部涂抹Pep-1-vMIP-II 2h后,用免疫组织化学方法测定不同组织器官中vMIP-II的穿膜情况。用实时定量PCR,检测24h后不同组织器官中ARGs mRNA表达... 详细信息

目的:探讨融合蛋白Pep-1-vMIP-II在影响宿主ARGs基因表达抵抗HIV-I感染中的作用。方法:小鼠背部涂抹Pep-1-vMIP-II 2h后,用免疫组织化学方法测定不同组织器官中vMIP-II的穿膜情况。用实时定量PCR,检测24h后不同组织器官中ARGs mRNA表达水平;用Graphpad Prism 5软件分析实验数据。结果:皮肤涂抹Pep-1-vMIP-II组2h后vMIP-II在肾脏分布明显增加(P<0.05);涂抹Pep1-vMIP-II 24h后,白细胞、肾、心、肝和肺中CCL5的mRNA表达水平有明显差异(P<0.05),脑组织中CCL5的表达未见明显增加;MX1、MX2在多种组织中都表达上调(P<0.05)。结论:Pep-1-vMIP-II可以穿透小鼠皮肤进入肾脏组织,主要通过激活CCL5基因表达发挥调控作用。

关键词： HIV-1 RNA 信使/遗传学白细胞肺基因表达皮肤肾心缩期小鼠

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RNA聚合酶Ⅱ中的CTD结构在mRNA转录和加工偶联过程中的重要作用

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遗传 2003年第1期25卷 102-106页

作者：韩玉波张飞雄首都师范大学生物系100037

RNA聚合酶Ⅱ最大亚基末端的羧基端结构域(CTD)是由以七氨基酸(-Tyr-Ser-Pro-Thr-Ser-Pro-Ser-)为重复单位的高度保守的重复序列组成的。因其结构与功能上的特殊性,它在近年来受到学术界的广泛关注。诸多的实验表明,该结构可通过与mRNA... 详细信息

RNA聚合酶Ⅱ最大亚基末端的羧基端结构域(CTD)是由以七氨基酸(-Tyr-Ser-Pro-Thr-Ser-Pro-Ser-)为重复单位的高度保守的重复序列组成的。因其结构与功能上的特殊性,它在近年来受到学术界的广泛关注。诸多的实验表明,该结构可通过与mRNA加工因子的相互作用而对其加工过程产生直接或间接的影响,在mRNA的转录和加工的偶联过程中具有重要的作用。本文即是根据近年来的实验结果对其结构和功能做出的综述性介绍。

关键词： RNA 信使/遗传学 RNA聚合酶Ⅱ/遗传学重复序列核酸转录遗传综述

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围生期甲低大鼠行为发育及其与海马AR mRNA表达的相关性研究

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浙江大学学报（医学版） 2008年第3期37卷 283-288页

作者：季钗黄新文杨荣旺赵正言浙江大学医学院附属儿童医院浙江杭州310003

目的:探讨围生期甲状腺功能低下对青春期不同性别大鼠行为发育的影响及其与海马AR mRNA表达的相关性。方法:孕SD雌鼠复制围生期甲低仔鼠模型雌雄各24只,随机各选雌雄12只为治疗组,于出生当日起每天腹腔注射T4(2μg/100 g)至断奶,其余为... 详细信息

目的:探讨围生期甲状腺功能低下对青春期不同性别大鼠行为发育的影响及其与海马AR mRNA表达的相关性。方法:孕SD雌鼠复制围生期甲低仔鼠模型雌雄各24只,随机各选雌雄12只为治疗组,于出生当日起每天腹腔注射T4(2μg/100 g)至断奶,其余为甲低组;另设正常对照组雌雄各12只。第60天进行旷场试验和避暗试验。行为评估结束后收集海马组织,用RT-PCR法测定AR mRNA水平。结果:在旷场试验中,甲低组大鼠跨格数较正常组增多,直立次数和排粪便数减少(P<0.05),治疗组雄性的跨格数未达正常(P<0.05);避暗试验:甲低组大鼠错误次数较正常组明显增多(P<0.01),雌性组少于雄性组(P<0.01),雄性组记忆潜伏期明显缩短(P<0.001),治疗组学习能力较甲低组明显增强,与正常组无显著性差异(P>0.05)。海马AR mRNA表达在甲低组雄性大鼠较正常组减少,治疗组与正常组无显著性差异(P>0.05)。雄性大鼠海马AR mRNA表达与跨格数呈负相关(r=-0.537,P=0.001),与避暗试验错误次数也呈负相关(r=-0.532,P=0.001),与潜伏期呈正相关(r=0.564,P=0.000)。结论:围生期甲状腺功能低下使青春期大鼠的行为发育改变,在雄性尤为明显,这可能与雄性大鼠海马AR mRNA表达尚未恢复正常有关,生后及时治疗能减轻这种损害。

关键词：甲状腺功能减退症受体雄激素海马行为动物记忆障碍 RNA 信使/遗传学

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Intergenic transcription and developmental remodeling of chromatin subdomains in the human β-globin locus

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MOLECULAR CELL 2000年第2期5卷 377-386页

作者： Gribnau, J Diderich, K Pruzina, S Calzolari, R Fraser, P Erasmus Univ Dept Genet & Cell Biol NL-3000 DR Rotterdam Netherlands Babraham Inst Lab Chromatin & Gene Express Cambridge CB2 4AT England

Gene activation requires chromatin remodeling complexes, which hyperacetylate histones and enable factor access;however, the targeting mechanisms leading to the establishment and maintenance of large, hyperacetylated DNase-sensitive chromatin domains are unknown. Recent work has shown that histone acetyltransferases are associated with RNA-pol II complexes, suggesting that transcription of chromatin plays a role in chromatin modification. Here we show the human P-globin locus is divided into three differentially activated chromatin subdomains. Large transcripts precisely delineate the active domains at key cell cycle points associated with chromatin transitions and remodeling. We identify an element that initiates these transcripts, located in a region required for chromatin activation. The results suggest that intergenic transcription is required for chromatin remodeling of chromosomal domains.

关键词：乙酰基转移酶类/代谢细胞周期染色质/代谢染色质/超微结构染色体/代谢基因重排珠蛋白类/遗传学组蛋白酰基转移酶肝/代谢基因座调控区模型遗传学 RNA聚合酶Ⅱ/代谢 RNA 信使/生物合成 RNA 信使/遗传学酿酒酵母蛋白质类转录激活人类

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ETL, a novel seven-transmembrane receptor that is Developmentally regulated in the heart - ETL is a member of the secretin family and belongs to the epidermal growth factor seven-transmembrane subfamily

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JOURNAL OF BIOLOGICAL CHEMISTRY 2001年第6期276卷 4150-4157页

作者： Nechiporuk, T Urness, LD Keating, MT Univ Utah Eccles Inst Human Genet Dept Human Genet Salt Lake City UT 84112 USA Univ Utah Eccles Inst Human Genet Dept Internal Med Div Cardiol Salt Lake City UT 84112 USA Univ Utah Eccles Inst Human Genet Howard Hughes Med Inst Salt Lake City UT 84112 USA

Using differential display of rat fetal and postnatal cardiomyocytes, we have identified a novel seven-trans-membrane receptor, ETL. The cDNA-predicted amino acid sequence of ETL indicated that it encodes a 738-aa protein composed of a large extracellular domain with epidermal growth factor (EGF)-like repeats, a seven-transmembrane domain, and a short cytoplasmic tail. ETL belongs to the secretin family of G-protein-coupled peptide hormone receptors and the EGF-TM7 subfamily of receptors. The latter are characterized by a variable number of extracellular EGF and cell surface domains and conserved seven transmembrane-spanning regions. ETL mRNA expression is up-regulated in the adult rat and human heart. In situ hybridization analyses revealed expression in rat cardiomyocytes and abundant expression in vascular and bronchiolar smooth muscle cells. In COS-7 cells transfected with Myc-tagged rat ETL, rat ETL exists as a stable dimer and undergoes endoproteolytic cleavage of the extracellular domain. The proteolytic activity can be abolished by a specific mutation, T455A, in this domain. In transfected mammalian cells, ETL is associated with cell membranes and is also observed in cytoplasmic: vesicles. ETL is the first seven-transmembrane receptor containing EGF-like repeats that is developmentally regulated in the heart.

关键词：选择性剪接氨基酸序列碱基序列 COS细胞染色体图染色体人 1对 DNA引物二聚作用表皮生长因子/遗传学基因表达调控发育期心脏/生长和发育分子序列数据心肌/代谢 RNA 信使/遗传学受体细胞表面/化学受体细胞表面/遗传学受体 G-蛋白偶联分泌素/遗传学序列同源性氨基酸增量调节动物人类大鼠

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Reverse transcription-PCR differential display analysis of Escherichia coli global gene regulation in response to heat shock

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APPLIED AND ENVIRONMENTAL MICROBIOLOGY 1999年第12期65卷 5386-5393页

作者： Gill, RT Valdes, JJ Bentley, WE Univ Maryland Dept Chem Engn College Pk MD 20742 USA Univ Maryland Inst Biotechnol Agr Biotechnol Ctr College Pk MD 20742 USA USA Edgewood Res Dev & Engn Ctr Aberdeen Proving Ground MD 21010 USA

A reverse transcription (RT)-PCR technique was developed to analyze global gene regulation in Escherichia coli. A novel combination of primers designed specifically for the start and stop regions of E. coli genes (based on the findings of Fislage et al. [R, Fislage, M, Berceanu, Y, Humboldt, M. Wendt, and N. Oberender, Nucleic Acids Res. 25:1830-1835, 1997]) was used as an alternative to the poly(T) primers often used in eukaryotic RT-PCR. The validity of the technique was demonstrated by applying it to heat shock analysis, Specifically, RT-PCR-amplified total RNA from heat-shocked and non-heat-shocked cells were hybridized with slot blots of the Kohara set (U, Kohara, K, Akiyama, and K. Isono, Cell 50:495-508, 1987;S. Chuang, D, Daniels, and F. Blattner, J, Bacteriol. 175:2026-2036, 1993), The signals obtained for heat-shocked and control cultures of each clone were compared, and differences in intensity were evaluated by calculating induction ratios. Clones that were considered significantly induced were subsequently mapped by the Southern blot technique in order to determine specific gene upregulation, Also, for several genes, Northern blotting and total RNA dot blotting were performed to confirm that the transcript levels in the original RNA samples were different. This technique extended previously described methods for studying global gene regulation in E, coli by incorporating a PCR amplification step in which global, mRNA-specific primers were used. In addition, the method employed here can be easily extended to study E. coli global gene regulation in response to additional environmental stimuli.

关键词：碱基序列印迹法 RNA 印迹法 DNA 染色体图/方法 DNA引物 DNA探针大肠杆菌/遗传学大肠杆菌/生长和发育发酵基因表达调控细菌 RNA 细菌/遗传学 RNA 信使/遗传学可重复性结果逆转录聚合酶链反应/方法

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Complexes of the G protein subunit Gβ₅ with the regulators of G protein signaling RGS7 and RGS9 -: Characterization in native tissues and in transfected cells

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JOURNAL OF BIOLOGICAL CHEMISTRY 2000年第32期275卷 24872-24880页

作者： Witherow, DS Wang, Q Levay, K Cabrera, JL Chen, J Willars, GB Slepak, VZ Univ Miami Sch Med Dept Mol & Cellular Pharmacol Miami FL 33136 USA Univ Miami Sch Med Program Neurosci Miami FL 33136 USA Univ So Calif Sch Med Dept Ophthalmol Los Angeles CA 90033 USA Univ Leicester Dept Cell Physiol & Pharmacol Leicester LE1 9HN Leics England

A novel protein class, termed regulators of G protein signaling (RGS), negatively regulates G protein pathways through a direct interaction with G alpha subunits and stimulation of GTP hydrolysis. An RGS subfamily including RGS6, -7, -9, and -11, which contain a characteristic G gamma-like domain, also has the unique ability to interact with the G protein beta subunit G beta(5). Here, we examined the behavior of G beta(5), RGS7, RGS9, and G alpha in tissue extracts using immunoprecipitation and conventional chromatography. Native G beta(5) and RGS7 from brain, as well as photoreceptor-specific G beta(5)L and RGS9, always co-purified as tightly associated dimers, and neither RGS-free G beta(5) nor G beta(5)-free RGS could be detected, Co-expression in COS-7 cells of G beta(5), dramatically increased the protein level of RGS7 and vice versa, indicating that cells maintain G beta(5) : RGS stoichiometry in a manner similar to G beta gamma complexes. This mechanism is non-transcriptional and is based on increased protein stability upon dimerization. Thus, analysis of native G beta(5)-RGS and their coupled expression argue that in vivo, G beta(5) and G gamma-like domain-containing RGSs only exist as heterodimers. Native G beta(5)-RGS7 did not co-precipitate or co-purify with G alpha(o) or G alpha(q);nor did G beta(5)-L-RGS9 with G alpha(t). However, in transfected cells, RGS7 and G beta(5)-RGS7 inhibited G alpha(q)-mediated Ca2+ response to muscarinic M3 receptor activation. Thus, G beta(5)-RGS dimers differ from other RGS proteins in that they do not bind to G alpha with high affinity, but they can still inhibit G protein signaling.

关键词：脑/代谢 COS细胞细胞膜/代谢二聚作用 GTP结合蛋白质β亚单位 GTP结合蛋白质类/代谢异源三聚体GTP结合蛋白质类/化学异源三聚体GTP结合蛋白质类/遗传学异源三聚体GTP结合蛋白质类/代谢氯醋甲胆碱/药理学蛋白质结合 RGS蛋白质类/化学 RGS蛋白质类/遗传学 RGS蛋白质类/代谢 RNA 信使/遗传学受体毒蕈碱M3 受体毒蕈碱/药物作用受体毒蕈碱/生理学重组蛋白质类/化学重组蛋白质类/代谢信号传导/药物作用信号传导/生理学转录遗传转染动物大鼠

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Muscle develops a specific form of small heat shock protein complex composed of MKBP/HSPB2 and HSPB3 during myogenic differentiation

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JOURNAL OF BIOLOGICAL CHEMISTRY 2000年第2期275卷 1095-1104页

作者： Sugiyama, Y Suzuki, A Kishikawa, M Akutsu, R Hirose, T Waye, MMY Tsui, SKW Yoshida, S Ohno, S Yokohama City Univ Sch Med Dept Biol Mol Kanazawa Ku Yokohama Kanagawa 2360004 Japan Chinese Univ Hong Kong Dept Biochem Shatin NT Peoples R China Kyoto Univ Grad Sch Med Dept Pathol & Tumor Biol Sakyo Ku Kyoto 6068501 Japan

Previously, we identified a new mammalian sHSP, MKBP, as a myotonic dystrophy protein kinase-binding protein, and suggested its important role in muscle maintenance (Suzuki, A., Sugiyama, Y., Hayashi, Y., Nyu-i, N., Yoshida, M., Nonaka, I., Ishiura, S., Arahata, K., and Ohno, S. (1998) J. Cell Biol. 140, 1113-1124). In this paper, we develop the former work by performing extensive characterization of five of the six sHSPs so far identified, that is, HSP27, alpha B-crystallin, p20, MKBP/HSPB2, and HSPB3, omitting lens-specific alpha A-crystallin. Tissue distribution analysis revealed that although each sHSP shows differential constitutive expression in restricted tissues, tissues that express all five sHSPs are only muscle-related tissues. Especially, the expressions of HSPB3, identified for the first time as a 17-kDa protein in this paper, and MKBP/HSPB2 are distinctly specific to muscles. Moreover, these sHSPs form an oligomeric complex with an apparent molecular mass of 150 kDa that is completely independent of the oligomers formed by HSP27, alpha B-crystallin, and p20. The expressions of MKBP/HSPB2 and HSPB3 are induced during muscle differentiation under the control of MyoD, suggesting that the sHSP oligomer comprising MKBP/HSPB2 and HSPB3 represents an additional system closely related to muscle function. The functional divergence among sHSPs in different oligomers is also demonstrated in several ways: 1) an interaction with myotonic dystrophy protein kinase, which has been suggested to be important for the maintenance of myofibril integrity, was observed only for MKBP/HSPB2;2) a myotube-specific association with actin bundles was observed for HSP27 and alpha B-crystallin, but not for MKBP/HSPB2;and 3) sHSPs whose mRNAs are induced by heat shock are alpha B-crystallin and HSP27. Taken together, the results suggest that muscle cells develop two kinds of stress response systems composed of diverged sHSP members, and that these systems work independently in muscle

关键词：载体蛋白质类/遗传学载体蛋白质类/代谢细胞分化细胞系晶体蛋白质类/遗传学基因表达调控基因文库 HSP27热休克蛋白质类热休克蛋白质类/遗传学热休克蛋白质类/代谢哺乳动物肌骨骼/细胞学肌骨骼/代谢器官特异性 RNA 信使/遗传学重组蛋白质类/代谢逆转录聚合酶链反应序列比对序列同源性氨基酸转录遗传转染成年人动物女(雌)性人类男(雄)性小鼠大鼠

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