基于PDA的嵌入式ECG监护分析仪

目的为了方便快捷地实时监测心律失常,设计一种基于PDA的嵌入式ECG监护分析仪。方法仪器功能包括低功耗MSP430系列单片机控制心电信号的采集、放大、滤波、转换和在PDA上实时显示、存储和分析,并实现心电数据的远程传输。对仪器工作原理进行分析,把监护分析仪总体结构划分为子模块,并具体介绍各子模块的功能与实现。结果与结论与医院现有的三导全自动心电图机、Holter动态心电图机进行对照,本监护分析仪携带方便、操作简单、接收的心电信号无失真,连续工作时间大于20h等。各高频干扰信号对ECG信号无影响;软件可对心电信号进行实时显示,可连续记录15h的原始心电数据。     

C3d-P28增强乙型肝炎病毒特异性基因免疫效果的研究

目的　观察补体C3d P2 8对基因免疫的调节作用及其不同拷贝数对调节作用的影响 ,为增强基因免疫效果寻求新方法。方法　PCR法获得补体C3d P2 8编码基因并以头尾串连方式将1～ 4拷贝C3d P2 8编码基因克隆至pVAON33,构建pVAON33 P2 8.[1～ 4 ]重组质粒 ,然后将HBV preS2 S编码基因分别插入pVAON33和pVAON33 P2 8.[1～ 4 ]质粒获得pVAON33 S2 S和pVAON33 S2 S P2 8.[1～ 4 ]重组质粒。肌肉注射各重组质粒DNA(每只 10 0 μg 10 0 μl)初次免疫小鼠 ,并以pVAON33为对照 ;12周后皮下注射HBsAg蛋白加强免疫各组小鼠 ,ELISA法检测免疫小鼠血清特异性抗 HBs IgG。结果　pVAON33 S2 S重组质粒免疫小鼠可诱导产生特异性抗 HBs IgG ,含不同拷贝C3d P2 8编码基因的重组质粒可诱导更高的特异性抗体 ,其中pVAON33 S2 S P2 8.4重组质粒诱导的抗体水平最高 (P <0 .0 1)。蛋白加强免疫后 ,含C3d P2 8编码基因重组质粒免疫组抗 HBs IgG迅速上升 ,并明显高于pVAON33 S2 S重组质粒免疫组 (P <0 .0 5 ) ,pVAON33 S2 S P2 8.4重组质粒诱导的抗体仍维持最高水平。结论　不同拷贝的C3d P2 8能不同程度地增强HBV preS2 S基因免疫诱导的特异性体液免疫及其蛋白加强后的回忆反应 ,其中 4拷贝C3d P2 8的增强作用较为显著。     

细胞间黏附分子1基因K469E多态性与冠状动脉粥样硬化性心脏病的关联研究

目的研究中国汉族人群中细胞间黏附分子1（intercellular adhesion moleculel，ICAM1）基因K469E多态性与冠状动脉粥样硬化性心脏病（简称冠心病）的关联。方法采用聚合酶链反应．限制性片段长度多态性方法检测了173例冠心病患者和141名对照的ICAM1基因K469E基因型和等位基因的分布。结果基因型频率符合Hardy-Weinberg平衡。冠心病组的KK基因型的频率显著高于对照组（64．2％比48．9％，P〈0．01），同样，冠心病组K等位基因的频率显著高于对照组（79．2％比69．9％，P〈0．01）。经Logistic回归分析排除年龄，性别，和冠心病其它危险因素的影响后，KK纯合子患冠心病的危险性是KE和EE基因型的2．35倍（95％CI：1．03-5．36，P〈0．05）。结论ICAM1基因K469E多态性与中国汉族人冠心病的危险性相关，其中K等位基因可能是冠心病的遗传危险因素。     

一个遗传性非综合征型耳聋家系的突变分析

目的分析一个遗传性非综合征型耳聋家系的突变，并探讨缝隙连接蛋白beta2(gap junction protein beta 2，GJB2)基因235delC突变是否会加重线粒体A1555G突变导致的非综合征型耳聋症状。方法对一个母系遗传性非综合征型耳聋核心家系72个成员取外周血提取DNA，经聚合酶链反应扩增后，利用Alw26Ⅰ限制性内切酶酶切及直接测序验证，对其线粒体DNA突变进行研究；利用ApaⅠ限制性内切酶酶切及直接测序验证，筛查核心家系中GJB2基因235delC突变情况，并对GJB2基因235delC和线粒体A1555G突变的关系进行研究。结果在27名母系成员中均发现具有线粒体A1555G突变，呈母系遗传；具有耳聋表型的为21人(77．8％)，家族外显率高；所筛查的包括配偶在内的72名个体中，仅3例具有GJB2基因235delC杂合子突变，且均出现在母系成员中，但3例的耳聋表型却不同。结论线粒体A1555G突变是本家系耳聋遗传易感性的基础，在该家系中GJB2基因的235delC杂合子突变未加重线粒体A1555G突变导致的非综合征型耳聋。     

DNA vaccination using bacillus Calmette-Guerin-DNA as an adjuvant to enhance immune response to three kinds of swine diseases

In order to enhance the immune efficacy of DNA vaccination, experiments were conducted to investigate the regulating effects of Bacillus Calmette-Guerin (BCG)-DNA as an adjuvant on immune responses of mice against foot-and-mouth disease (FMD), Aujeszky's disease (AjD) and classical swine fever (CSF). BCG-DNA was purified from BCG by ion-exchange chromatography. Three DNA vaccines (pVSG, pVgD and pVE2) against the respective infection were constructed, and BCG-DNA was coimmunized to mice by muscle injection. The results showed that titres of specific immunoglobulin (Ig)G to the vaccines mounted remarkably in the sera of the adjuvant covaccinated mice (P < 0.01). Antibody isotype IgG2a and IgG1 also increased, respectively, in mice coimmunized with BCG-DNA compared with those of the control groups (P < 0.01). Cellular immune cytokine interferon-gamma and cytotoxic T lymphocytes were detected in coimmunized BCG-DNA groups (P < 0.05). Whereas interleukin-4, humoral immune cytokine, was not significant (P > 0.05). These results suggest that codelivery of BCG-DNA with DNA vaccines against FMD, AjD and CSF can enhance the induction of antigen-specific, especially, cell-mediated immunity.     

抗α-玉米赤霉醇单克隆抗体的研制及初步应用

目的:制备抗α-玉米赤霉醇(α-ZER)的单克隆抗体(mAb),建立对动物源性食品中残留α-ZER及其同系物的检测方法。方法:结合O-羧甲基羟胺法和EDC法制备α-ZER-BSA偶联物,免疫BALB/c小鼠,采用杂交瘤技术制备mAb。用夹心ELISA测定mAbIg亚类;按Beatty法计算亲和常数;间接ELISA法测定效价;用直接竞争ELISA检测mAb与α-ZER的同系物、己烯雌酚、19-去甲睾酮、链霉素及氯霉素等的交叉反应;绘制α-ZER标准品竞争抑制曲线,检定抗体的灵敏度。用该抗体建立的直接竞争ELISA对37份动物肝组织样品进行检测,并与HPLC检测结果比较。结果:紫外线扫描证明,α-ZER-BSA偶联成功。实验获得8株可稳定分泌抗α-ZERmAb杂交瘤细胞株,其中1株(4E5)分泌的mAb效价较高,为5.142×107,其Ig亚型为IgG1。该mAb能特异识别α-ZER及其同系物,而与其它常用兽药无交叉反应。建立了α-ZER直接竞争ELISA,从HPLC确认为阴性的37份样品中,筛出8份阳性样品。结论:利用mAb建立的直接竞争ELISA检测方法,适合动物源性食品中残留α-ZER及其同系物的快速筛选。     

注射型藻酸钙/BMP复合材料的制备及成骨性能研究

研究藻酸钙（CaAG）／BMP-2复合材料的成骨性能，并观察骨髓基质细胞（BMSC）的加入对成骨性能的影响。采用藻酸钠、骨形态发生蛋白-2（BMP-2）和氯化钙（CaCl2）混合制备藻酸钙／BMP复合材料，以加有骨髓基质细胞的该材料为对照，进行体外观察和肌袋试验，通过用X射线、ALP染色、Von Kossa染色、HE染色、新生骨计量等方法，研究其诱导成骨活性。CaAG-BMP组和CaAG-BMP—BMSC组均在4～6周形成新生骨组织，两组标本均为编织骨形态．X线检查、AKP染色、Von Kossa染色、HE染色提示成骨过程相似。两组在新生骨计量上无显著性差异。藻酸钙／BMP复合材料可在肌肉环境下有效形成一定体积的新生骨组织；应用骨骼肌异位诱导成骨原理，该复合材料在成骨过程中无需加入外源性的种子细胞。     

Holoprosencephaly: from Homer to Hedgehog

Holoprosencephaly (HPE), a common developmental defect affecting the forebrain and face, is etiologically heterogeneous and exhibits wide phenotypic variation. Graded degrees of severity of the brain malformation are also reflected in the highly variable craniofacial malformations associated with HPE. In addition, individuals with microforms of HPE, who usually have normal cognition and normal brain imaging, are at risk for having children with HPE. Some obligate carriers for HPE may not have any phenotypic abnormalities. Recurrent chromosomal rearrangements in individuals with HPE suggest loci containing genes important for brain development, and abnormalities in these genes may result in HPE. Recently, Sonic Hedgehog (SHH) was the first gene identified as causing HPE in humans. Proper function of SHH depends on cholesterol modification. Other candidate genes that may be involved in HPE include components of the SHH pathway, elements involved in cholesterol metabolism, and genes expressed in the developing forebrain.