Comparative study of large yellow croaker (Pseudosciaena crocea) and marine medaka (Oryzias melastigma) Hepcidin gene expression and peptide functions

抗菌肽是一类从生物体中分离的小分子多肽，具有抗细菌，真菌，病毒，肿瘤和寄生虫等多种作用。由于抗菌肽来源于生物体本身，同时病原菌不易对其产生耐药性，所以被视为一种极有潜力的抗生素的替代品。Hepcidin抗菌肽是鱼类抗菌肽家族中的重要一种，鱼体内的Hepcidin更是鱼类先天免疫因子的重要组成部分。本研究运用分子生物学技术对养殖鱼类大黄鱼（Pseudosciaenacrocea）Hepcidin和模式动物海水青鳉（Oryziasmelastigma）Hepcidin的基因表达特性与功能进行了比较研究和探讨，主要研究成果如下： 1. 克隆获得了源于海水养殖大黄鱼Hepcidin（Pc-hepc）...Antimicrobial peptides (AMPs) are a group of small peptides amplified from organisms which have anti-bacterial activity, anti-fungi activity, anti-virus activity, anti-tumors activity, etc. AMPs are thought to be very good substitutes for antibiotics that they can defense pathogens invading with less drug tolerance for they are derived from organism themselves. Hepcidin is an important AMP family,...学位：理学博士院系专业：海洋与环境学院环境科学与工程系_环境科学学号：2262007015392

Prevention for Seismic Secondary Geological Disasters and Aseismic nalysis for Rural and School Buildings in Xiamen City

2008年5月12日，我国四川省汶川县发生里氏8.0级特大地震。此次大地震造成了大量的房屋破坏，农村房屋的破坏最为严重。农村房屋抗震能力低下、抗震措施不足是最主要的原因。学校建筑在这次地震中也受到不同程度的破坏，作为公共性建筑和避难场所，这样的破坏会造成很大的损失，其根本的原因是学校建筑结构体系的缺陷。除了以上两种原生灾害外，地震次生地质灾害所带来的破坏也极为严重。 地质情况不佳、农村房屋抗震性能差、学校在役建筑抗震能力低恰恰是厦门地区存在的抗震薄弱环节，如果地震来临，也将遭受严重的破坏，厦门市这几个方面的系统研究还比较少，为此，本文针对这三个方面做了深入的研究和探讨。 本文首先分析了地震...A destructive earthquake with the seismic magnitude of 8.0 degree occrued in May 12 2008, known as Wenchuan earthquake in Sichuan Province. This severe earthquake led to the collapse and severe damage to huge numbers of buildings, especially the rural houses. The main reason is lower seismic defence capacity and deficient seismic measures. As a public building and evacuee shelter, school buildings...学位：工学硕士院系专业：建筑与土木工程学院土木工程系_结构工程学号：2532007115219

论我国对外反倾销实践的特点及存在的问题

反倾销是保护国内产业健康发展,维护经济安全的一个重要手段。本文分析了入世后中国对外反倾销的特点,并分析了现存反倾销实践中所存在的问题。最后,笔者对如何完善反倾销实践及应注意的问题,提出了自己的看法

典型框架式学校建筑的抗震性能分析

本文在研究汶川地震灾区学校建筑震害的基础上,探讨了中小学校典型框架结构即外廊悬挑式单跨框架结构的抗震性能。基于静力非线性方法,用ETABS对典型框架结构进行弹塑性分析,对不同水准下结构的出铰顺序,出铰数量,侧向位移等进行对比,得出相应的抗震性能评价,本文还对该框架结构的薄弱部位进行分析,提出加固改造的相关建议。本文的研究工作对提高学校建筑的抗震能力具有实用价值

提高闽南地区农村房屋抗震能力的探讨

本文从农村房屋震害分析入手,针对闽南地区农村房屋结构特点及在抗震设防中存在的问题,提出了改善农村房屋抗震性能的具体措施。本文还就新建农村房屋的抗震设防及既有农村房屋的加固改造提出相关建议

地震次生地质灾害的分析及防治对策

本文基于对5.12汶川大地震灾后现场灾情以及灾后抢险、恢复重建工作等方面的调查,分析了地质灾害及造成这些地质灾害的原因,并通过分析,提出防治地质灾害的措施。本文还就减轻地质灾害、提高规划选址水平、增强防灾意识等提出建议,供今后在工程建设时参考

Relative Quantification of Hepcidin Gene Transcripts in Pseudosciaena crocea

以大黄鱼(PSEudOSCIAEnA CrOCEA)管家基因18S rrnA和β-ACTIn作为内参基因,分别比较2个内参基因建立的相对定量曲线,最终确立以18S rrnA为参比基因,定量分析大黄鱼的HEPCIdIn抗菌肽基因.该相对定量分析方法所得结果与nOrTHErn-blOT方法一致.应用建立的HEPCIdIn基因的实时荧光相对定量研究方法,对大黄鱼头肾中的HEPCIdIn基因转录物进行相对定量,为今后开展鱼体内免疫相关基因的表达特性、诱导机制等工作奠定研究基础.同时,克隆得到的大黄鱼β-ACTIn基因和18S rrnA基因片段已提交基因库,并获得登录号.Two house-keeping genes,18S rRNA and β-actin,were cloned as endogenous genes for relative quantification of an antimicrobial peptide hepcidin in large yellow croakers(Pseudosciaena crocea) using the real-time RT-PCR.In comparision of the slopes of the relative standard curves between the two endogenous genes and the target gene,a relative quantification method for hepcidin gene using 18S rRNA as the calibrator gene was established.Results obtained with this method were consistent with the rusults from Northern-blot,suggesting feasibility and effectiveness of the method.This paper provides an effective method to investigate expression patterns and induction profiles of the hepcidin gene in large yellow croakers,and other immunity-related genes in fish bodies.The cloned fragments of 18S rRNA and β-actin genes from the large yellow croaker have been submitted to the GenBank,which have provided accession numbers.福建省科技计划重点资助项目(2007I0022);上海市高校选拔优秀青年教师科研专项基金资助项目(SHU08021);上海大学创新基金资助项目(A10011109006

Construction of SSH library from haemocyte of variously colored abalone challenged with bacteria and differential expression analysis of macrophage expressed protein

以雌性杂色鲍为对象,以大肠杆菌、副溶血弧菌、溶壁微球菌、表皮葡萄球菌、金黄色葡萄球菌的混悬液做为攻毒菌,利用抑制性差减杂交(SSH)技术构建细菌攻毒的杂色鲍血淋巴细胞SSHcDNA文库。随机挑取生长菌落110个克隆子,进行菌液PCR鉴定,计算文库重组率为98.18%,文库容量为1.37×106pfu。将重组子测序,经BLAST一致性搜索比对分析,有一重组片段含有穿孔素(Perforin)保守结构域,为巨噬细胞表达蛋白(MEP)类穿孔素部分cDNA序列,片段大小为1551bp,连续编码517个氨基酸残基,申请GenBank登录号为EU272049。经半定量PCR和荧光定量PCR差异显示分析,发现在细菌感染状态下MEP基因在血淋巴细胞中存在明显的上调表达现象。Abalones are considered to be the most precious delicacy from the sea, and become very important commercial seafood in aquaculture worldwide. Variously colored abalone (Haliotis diversicolor Reeve, 1846) has been widely cultured on the southeast coast for more than twenty years. However, abalone culture frequently suffers from bacterial infection and mass mortality of reared abalones causes serious economic losses. Unfortunately, knowledge of the defense mechanism in this animal is still lacking. In this study, using suppression subtractive hybridization (SSH) technology, a forward SSH li-brary was constructed from haemocytes of H. diversicolor, with the content of 1.37×106 pfu and the recombinant rate of98.18%. After the recombinant plasmids were sequenced, partial cDNA of macrophage expressed protein (MEP) was recognized based on BLAST searches in NCBI, with the size of 1 551 bp, and continuously encoding 517 amino acids. Semi-quantitative PCR and quantitative real-time PCR results showed that MEP cDNA was distinctly up-regulated in haemocytes of the bacterial-challenged group compared to the unchallenged group. The gene information obtained from this library will provide new insights into the immune mechanism of H. diversicolor and facilitate future study of target genes involved in the response to invading microorganisms.国家高技术研究发展计划项目(863计划)(编号:2007AA091406)资助~

生育酚结合蛋白通过磷酸肌醇3激酶途径抑制前列腺癌的生长

【目的】探讨生育酚结合蛋白（TAP）对前列腺癌细胞的生长调节及其分子机制。【方法】四甲基偶氮唑盐生长试验（MTY）测定细胞增殖情况，体外集落形成试验测定各细胞的致癌能力，高效液相层析法检测细胞内维生素E的浓度，利用基因转染、基因沉默、免疫印迹、Northem blot，RT-PCR、荧光定量PCR、免疫沉淀等方法研究TAP对前列腺细胞生长的作用及机制。【结果】TAPmRNA水平在前列腺癌细胞LNCaP、PC-3、DU-145、CWR22R中均较正常前列腺细胞RWPE-1低。TAP可促进癌细胞保留维生素E并增强其抗前列腺癌增殖的效应。无维生素E作用下．转染TAP可抑制前列腺癌细胞LNCaP、DU-145的生长，第6天细胞数比对照组分别减少35．8％与42．4％；LNCaP细胞克隆形成率下降54．3％（P〈0．05）。利用siRNA在良性前列腺细胞HPr-1中沉默TAP基因，培养9d后，细胞数比对照组增加124．3％（P〈0．01）。TAP通过抑制磷酸肌醇P13激酶信号．而非通过影响细胞周期或雄激素受体信号发挥作用。免疫沉淀实验表明TAP通过抑制P13K的亚单位p110与p85的相互作用，进而干扰P13K-Akt信号通路；持续激活Akt的活性可削弱TAP对前列腺癌细胞生长的抑制能力。【结论】TAP不仅能促进前列腺癌细胞摄取和增强维生素E的抗前列腺癌效应，还可通过非维生素E途径发挥作用．可能是防治前列腺癌的有价值的分子靶点