益气清热方对幽门螺杆菌ureA基因表达影响的研究

目的：观察益气清热方对幽门螺杆菌（Hp）urea基因表迭的影响。方法：将益气清热方、黄芪及黄芩按Hp最大耐受浓度加入培养基进行Hp液体培养，72小时后提取RNA，应用实时荧光RT-PCR方法定量检测HpureA基因mRNA。结果：与对照组相比，黄芪和复方组HpureA基因mRNA水平低于对照组，有显著性差异（P〈0．001），而黄芩组与对照组相比无显著差异。结论：益气清热方和黄芪对HpureA基因的表达有下调作用。     

分子信标实时定量PCR法检测胃癌中Survivin基因mRNA的表达

目的 采用分子信标实时定量PCR方法检测胃癌样本中Survivin基因mRNA的表达量，并分析其与临床资料之间的关系。方法 设计Survivin基因特异性分子信标探针和引物，以cDNA克隆重组质粒为标准品，建立Survivin基因实时定量检测方法；并检测胃癌样本中Survivin基因mRNA的模板拷贝数。结果 Survivin基因mRNA分子信标实时定量检测方法的线性范围为10^3-10^10拷贝数，批间变异为28．16％，批内变异为13．34％，灵敏度为42个拷贝数，平均回收率为109．83％；胃癌组织、癌旁组织和正常组织以及胃良性病变组织间比较，mRNA模板拷贝数差异有显著性（P〈0．05）；胃癌样本中mRNA模板拷贝数与淋巴结转移和2年生存期以及病理类型相关（P〈0．05）。结论 成功建立Survivin基因分子信标实时定量检测方法。胃癌中该基因表达的模板拷贝数可作为预测淋巴结转移、评价预后的一个重要指标。     

护理质量时间-行为位点监控考核应用研究

目的通过护理时间-行为程序实时动态监控，使护理工作落实到位，护理质量提高，有效减少护理纠纷，提高患者满意率。方法设计22个监控考核点，时间位点11个，行为位点11个，对全院35个临床科室全面实时监控。结果病区执行监控后各项护理质量考核总平均分值明显高于执行前；患者满意率均有所提高，其中患者满意率提高明显，实施护理时间-行为位点监控前的护理纠纷投诉率31．96％，患者满意度88．15％，到实施后的纠纷投诉率降低为4．12％，患者满意度提高到98．61％。其结果差异有显著性。结论本研究采用的护理质量时间-行为位点监控是一项高效，便于临床操作的双向评估、双向监督的管理模式，值得推广运用。     

星形胶质细胞毒蕈碱样乙酰胆碱受体M1、M3、M5亚型基因克隆及序列分析

目的克隆胶质细胞M1、M3、M5受体亚型基因序列,比较胶质细胞和神经细胞M1、M3、M5受体亚型基因序列、蛋白质序列的差异。方法根据神经细胞M1、M3、M5受体基因序列全长设计特异性探针,采用RT-PCR方法扩增胶质细胞M1、M3、M5受体亚型基因序列,并对其进行克隆测序。结果测序得到胶质细胞M1、M3、M5受体亚型基因序列,与神经细胞比较,M1受体差异碱基4个,发生氨基酸改变的有1个;M3受体差异碱基有8个,发生氨基酸改变的位点有4个;M5受体差异碱基1个,发生氨基酸改变的有1个。结论胶质细胞与神经细胞M1、M3、M5受体亚型在基因和蛋白序列上具有明显差异。     

实时荧光定量RT-PCR检测内皮细胞t-PA mRNA方法的建立

目的建立实时RT-PCR检测人微血管内皮细胞组织型纤溶酶原激活物(t-PA)mRNA的表达。方法提取人微血管内皮细胞总RNA,经RT-PCR获得靶基因(t-PA)及管家基因(β-actin)的PCR产物。纯化后,作为标准品梯度稀释,采用SYBR G reen I定量PCR检测,建立标准曲线。方法学考核参数为特异性、线性范围、精密度和重复性。分析全反式维甲酸对内皮细胞表达t-PA mRNA的干预效果。结果定量方法特异性好,检测的灵敏度达103拷贝,线性范围为103~1010拷贝,循环阈值与PCR体系中起始模板量的对数值之间有着良好的线性关系(r2>0.990),批内变异≤3.10%,批间变异≤4.93%。1.25~20.00μmol.L-1的全反式维甲酸能明显上调内皮细胞t-PA mRNA的表达(P<0.01),且呈剂量依赖性。结论实时荧光定量RT-PCR的方法可对t-PA mRNA的表达进行准确定量,有助于溶栓药物药理学研究和新药筛选。     

上皮性卵巢癌中nm23-H1基因突变

目的　本文研究nm2 3 H1基因在上皮性卵巢肿瘤中的突变情况 ,以期寻找监测卵巢癌转移并能指导治疗的指标。方法　利用非同位素PCR SSCP技术研究nm2 3 H1基因的 5个外显子的突变情况。结果　正常卵巢、良性卵巢癌及交界性瘤均未发现突变 ,卵巢癌突变率显著增高。低分化程度卵巢癌突变率高于高、中分化程度的卵巢癌。浆液性卵巢癌nm2 3 H1基因突变率高于其他组织学类型。Ⅲ、Ⅳ期卵巢癌突变率高于Ⅰ、Ⅱ期卵巢癌。发生淋巴结转移的卵巢癌突变率高于无淋巴结转移的卵巢癌。结果　nm2 3 H1基因突变容易发生于分化程度较低、有淋巴结转移的晚期卵巢癌病例中。     

A molecular approach (multiplex polymerase chain reaction) for diagnosis of rhinosporidiosis

A New multiplex PCR have been developed in our laboratory using primer sets, aiming amplification of both D.N.A target fragments obtained in 18S RNA of commonly encountered fungi in human being and in Rhinosporidium seeberi using F1-fw/F2-rev (500 bp target) and Rhino-fw/ Rhino-rev (.177 bp target). This multiplex PCR has been found to be able to delect R. seeberi from clinical samples and differentiate it from other fungi. Furthermore, by this multiplex PCR, R. seeberi, phylogenitically appears to belong to a member of so called DRIPs clade of fish parasite not a cyanobacterium as claimed previously, by some workers.     

250例非淋菌尿道炎诊治体会

非淋菌尿道炎是相对于淋菌性尿道炎的一种高发性传播疾病。其中绝大部分是由砂眼衣原体和解脲支原体所引起，属中医“淋症”范畴，病机多为湿热下注所致。笔者采用自拟通淋合剂配合四环素治疗，疗程平均二周，取得了痊愈率94％，有效率100％的显著疗效，值得推广应用。     

Trajectory reconstruction from trace evidence on spent bullets. II. Are tissue deposits eliminated by subsequent impacts?

STR-based individualisation of biological deposits on bullets after perforation of tissue, can identify the person injured or killed by a particular bullet and comparison with the firearms used can identify the weapon and thus possibly the person who did the shooting. In this study, the effect of subsequent impacts on intermediate targets such as loss of cells was investigated by amplification of mitochondrial (mt) DNA. Bovine tissue was perforated and the 9 mm Luger FMJ bullets were recovered from the bullet collector. The mt cytochrome-b (cyt-b) gene could be amplified by the polymerase chain reaction (PCR) from 14 out of 15 bullets. Examination with a scanning electron microscope (SEM) and an energy-dispersive X-ray spectrometer (EDS) demonstrated the presence of minute dried tissue deposits on all bullets (n = 10) but was not able to establish preferential locations. In a series of 25 gunshots, various intermediate targets (glass, wood, car metal, gypsum board, asphalt) were perforated/ impacted following perforation of tissue and the cyt-b gene could be typed from all bullets. It is concluded that subsequent impacts on intermediate targets do not eliminate enough biological deposits to render DNA analysis impossible and that the amplification of mtDNA is a useful additional method. Received: 5 September 2000 / Accepted: 25 November 2000