学龄前儿童末梢全血中微量元素含量检测结果分析

目的:了解厦门地区学龄前儿童末梢全血中5种微量元素水平及其与年龄、性别的关系。方法:采用原子吸收分光光度法测定厦门地区0~7岁儿童末梢全血中5种微量元素铜、锌、钙、镁、铁的含量,对其结果进行统计学分析。结果:学龄前儿童缺锌和缺铁的比例较高;不同年龄组儿童锌和铁的含量具有显著性差异,低年龄组的含量低于高年龄组,且婴儿期的锌和铁的含量明显低于大龄儿童;不同性别组儿童除血钙含量有显著性差异外,其他元素的含量无显著性差异。结论:不同年龄、性别、不同生活习惯都可能引起儿童体内微量元素含量的不同,建议根据不同情况对儿童尤其是婴幼儿进行微量元素缺乏的防治工作。     

单纯性肥胖儿童血清IGF-Ⅰ和血脂变化及相关性研究

目的:探讨单纯性肥胖儿童(SOC)血清胰岛素样生长因子(IGF-Ⅰ)和血脂水平及两者相关性。方法:单纯性肥胖儿童和正常对照组儿童各50例,应用常规生化检验方法检测其总胆固醇(TC)、甘油三脂(TG)、低密度脂蛋白胆固醇(LDL-C)及高密度脂蛋白胆固醇(HDL-C)等,用ELISA法测定IGF-Ⅰ水平。结果:与健康儿童相比,单纯性肥胖儿童血清IGF-Ⅰ、TG、TG、LDL-C及ApoB均明显高于对照组(P<0.05),而HDL-C、ApoAI均明显低于对照组(P<0.05)。血清IGF-Ⅰ的浓度与血脂TC、TG、LDL-C及ApoB呈负相关关系(P<0.01),与HDL-C、ApoAI呈正相关关系(P<0.01)。结论:血清IGF-Ⅰ浓度的改变与单纯性肥胖儿童血脂代谢紊乱存在显著相关性。     

厦门地区婴幼儿腹泻诺如病毒的基因分型研究

目的:了解厦门地区婴幼儿腹泻诺如病毒(Norovirus,NV)分子流行病学特征。方法:收集2010年5月-2011年4月全年轮状病毒抗原阴性的急性非细菌性腹泻标本,用实时荧光RT-PCR法扩增NV ORF1-ORF2结合区,测序扩增片段后进行比对、分型及构建系统进化树。结果:全年共获得366份标本,共检测出81份为NV阳性标本,其中GI组5例,GII组76例,NV在厦门地区全年可见散发,但在每年7月和11月有明显的流行高峰,且腹泻感染患儿主要集中在3岁以下患儿,已分型的NV基因组(55例)以GII-4/2006b占绝对优势,其构成比达60.00%(33/55),其余为GII-3占25.45%(14/55),GII-6占5.45%(3/55),GII-2占3.64%(2/55),此外GII-12、GI-4、GI-5各占1.81%(1/55)。结论:厦门地区NV夏秋季发病率较高,GII-4/2006b变异株为优势株,且感染的NV可能存在病毒重组。     

厦门市3075例儿童血铅检测与分析

目的：了解厦门市区儿童血铅水平分布状况,为预防儿童铅中毒提供依据。方法：随机选择我院门诊就诊的儿童3 075例,年龄0～12岁,原子吸收石墨炉法测定血铅值。结果：儿童血铅均值为64.98 g/L,铅中毒检出率为9.33%,随着年龄增加血铅均值及铅中毒检出率呈现逐渐增高的趋势,男童血铅均值及铅中毒检出率高于女童。结论：厦门市儿童铅中毒率接近国内其他城市平均水平,但血铅水平有随年龄增长而增加的趋势,铅污染对儿童身体健康危害应引起重视。     

子宫内膜异位症相关性不孕患者的子宫内膜差异表达基因cDNA文库的构建和筛选

目的:筛选子宫内膜异位症相关性不孕患者在着床窗口期子宫内膜差异表达基因,以了解子宫内膜容受性形成的分子基础。方法:采用抑制性消减杂交技术(SSH),建立人子宫内膜着床窗口期基因表达的消减文库,并通过测序和同源性比较确定差异表达的基因;用RT-PCR技术验证其中的翻译控制肿瘤蛋白(TCTP1)基因、载脂蛋白D(ApoD)基因、促红细胞生成素(EPO)基因在分泌期中期子宫内膜中的表达。结果:对消减文库中20个差异表达的克隆进行测序与同源性比较,筛选出8个在分泌期中期有差异表达的基因。其中TCTP1、ApoD、EPO基因在分泌期中期的表达水平有不同程度的升高,分别为1.35±0.12、1.24±0.10、1.05±0.23,分别与正常对照组的1.05±0.23、0.75±0.16、0.68±0.08比较,差异均有统计学意义(P<0.05)。结论:应用SSH可有效地筛选出内异症相关性不孕患者着床窗口期子宫内膜差异表达基因,为进一步研究子宫内膜容受性形成的分子机理提供新的线索。     

Construction, Expression and Identification of a Recombinant BCG Vaccine Encoding Human Mycobacterium Tuberculosis Heat Shock Protein 65

Heat shock protein 65 (HSP65) is one of the most important protective immunogens against the tuberculosis infection. The signal sequence of antigen 85B and the whole HSP65 DNA se-quence of human Mycobacterium tuberculosis (M. tuberculosis) were amplified from BCG genome and plasmid pCMV-MTHSP65 respectively by polymerase chain reactions (PCR). These two se-quences were cloned into the plasmid pBCG-2100 under the control of the promoter of heat shock protein 70 (HSP70) from human M. tuberculosis, yielding the prokaryotic shuttle expression plas-mid pBCG-SP-HSP65. Results of restriction endonuclease analysis, PCR detection and DNA se-quencing analysis showed that the two cloned DNA sequences were consistent with those previously reported, and the direction of their inserting into the recombinant was correct and the reading frame had been maintained. The recombinants were electroporated into BCG to construct the recombinant BCG vaccine and induced by heating. The induced expression detected by SDS-PAGE showed that the content of 65 kD protein expressed in recombinant BCG was 35.69% in total bacterial protein and 74.09% in the cell lysate supernatants, suggesting that the recombinant HSP65 gene could express in BCG with high efficiency and the expressed proteins were mainly soluble. Western-blot showed that the secretive recombinant proteins could specifically combine with antibody against M.tuberculosis HSP65, indicating that the recombinant proteins possess the biological activity of HSP65.     

Expression and Purification of SARS Coronavirus Membrane Protein

To construct a recombinant plasmid Pet23a-M, the gene encoding severe acute respiratory syndrome (SARS) coronavirus membrane protein was amplified by RT-PCR and cloned into the expression plasmid Pet23a. Results of restriction endonuclease analysis, PCR detection and DNA sequencing analysis revealed that the cloned DNA sequence was the same as that reported. The recombinants were transformed into Escherichia coli (E. Coli) BL21 (DE3) and induced by Isopropyl-β-D-thiogalactopyranoside (IPTG). The expression of 27 kD (1 kD=0. 992 1 ku) protein was detected by SDS-PAGE and pured by metal chelated chromatography. Results of Western-blot showed that this expressed protein could react with antibodies in sera of SARS patients during convalescence. This provided the basis for the further study on SARS virus vaccine and diagnostic agents.     

替罗非班在糖尿病合并冠脉三支病变患者PCI 术后的应用

目的：探讨替罗非班在糖尿病合并冠脉三支病变经皮冠状动脉介入治疗患者中的效果及安全性。方法选择120例接受经皮冠状动脉介入治疗的糖尿病合并冠脉三支病变患者作为临床观察对象,随机分为替罗非班组（n＝64）与对照组（n＝56）,对照组常规用药,替罗非班组在常规用药基础上予替罗非班治疗。比较两组在术后即刻TIMI血流、校正TIMI帧数、术后48 h的血小板计数和血小板集聚率与术后30 d的左室射血分数（LVEF）、左室舒张末期内径（LVEDD）、左室收缩末期内径（LVESD）;同时观察术后30 d的主要不良心血管事件（MACE）发生率和出血事件发生率。结果替罗非班组术后TIMI血流改善显著,达TIMI 3级血流明显高于对照组,术后校正TIMI帧数及术后48 h血小板集聚率均低于对照组（ P＜0.05）;替罗非班组术后30 d LVEF高于对照组（P＜0.05）, LVEDD、LVESD均低于对照组,两组比较差异均有统计学意义（P＜0.05）;术后30 d,替罗非班组心绞痛、心肌梗死、MACE发生率显著降低,与对照组比较差异有统计学意义（ P＜0.05）;两组出血事件发生率比较差异无统计学意义。结论对于需要经皮冠状动脉介入术治疗的糖尿病合并冠脉三支病变患者,合理使用替罗非班,能够有效提高心肌灌注水平,保护心功能,降低MACE的发生率,且不增加出血事件发生率,值得在临床中加以推广。     

人结核杆菌热休克蛋白65和Ag85A基因真核双表达质粒的构建和体外表达

目的构建人结核杆菌热休克蛋白65（Hsp65）与Ag85A基因的共表达载体pIRES-Hsp65-Ag85A,并检测其在体外的表达。方法利用聚合酶链反应（PCR）法及基因重组技术,以人结核杆菌基因组DNA为模板,扩增获得Hsp65与Ag85A的全长基因,并将其构建到真核表达质粒pIRES中获得共表达质粒pIRES-Hsp65-Ag85A。将该质粒转染Hela细胞,通过RT-PCR的方法检测目的基因的表达。结果经过测序证实重组质粒构建成功,该质粒在体外转染Hela细胞后可表达Hsp65与Ag85A两者的mRNA。结论成功构建了共表达质粒pIRES-Hsp65-Ag85A,该质粒能在人子宫颈癌细胞中表达。     

Construction and Expression of Bivalent Membrane-anchored DNA Vaccine Encoding Sj14FABP and Sj26GST Genes

In order to construct a eukaryotic co-expression plasmid containing membrane-anchored Sjc14FABP and Sjc26GST genes and identify their expression in vitro, Sj 14 and Sj26 genes were obtained by RT-PCR with total RNA of Schistosoma japonicum adult worms as the template and cloned into eukaryotic expression plasmid pVAC to construct recombinant plasmids pVAC-Sj14 and pVAC-Sj26. Then a 23 amino-acid signal peptide of human interleukin-2 (IL-2) upstream Sj14 or Sj26 gene and a membrane-anchored sequence containing 32 amino-acids of carboxyl-terminal of human plaeental alkaline phosphatase (PLAP) downstream were amplified by PCR as the template of plasmid pVAC-Sj14 or pVAC-Sj26 only to get two gene fragments including Sj14 gene and Sj26 gene. The two modified genes were altogether cloned into a eukaryotic co-expression plasmid pIRES, resulting in another new recombinant plasmid pIRES-Sj26-Sj 14. The expression of Sl14 and Sl26 genes was detected by RT-PCR and indirect immunofluorescent assays (IFA) when the plasmid pIRES-Sj26-Sj14 was transfected into eukaryotic Hela cells. Restriction enzyme analysis, PCR and sequencing results revealed that the recombinant plasmids pVAC-Sj14, pVAC-Sj26 and pIRES-Sj26-Sj 14 were successfully constructed and the expression of modified Sj 14 and Sj26 genes could be detected by RT-PCR and IFA. A bivalent membrane-anchored DNA vaccine encoding Sj14 and Sj26 genes was acquired and expressed proteins were proved to be mostly anchored in cellular membranes.