土壤环境基因组技术及其在新药发现中的应用

土壤是微生物最重要的生境，土壤微生物具有极大的多样性。然而传统培养技术只能得到约1％的微生物纯培养，其余都是未被纯培养微生物。利用土壤环境基因组技术，可以把未被纯培养微生物的基因克隆到载体中并在宿主中进行表达，获得比已培养微生物丰富的代谢产物多样性，这为我们发现新颖结构先导化合物以及新药开辟了新的道路。本文介绍应用环境基因组技术构建土壤DNA文库的思路和方法。     

医院图书馆与医务人员卫生法学教育

为了改变卫生人员和医学生的知识结构,自觉执行卫生法,培养适应新时代有道德、有文化、守纪律、高素质的卫生人员,必须要加强学习卫生法学教育。文章阐述了卫生法的概念,及其教育的意义,卫生法学在医疗实践中的作用;同时提出了图书馆如何配合医院开展卫生法学教育的措施,扩大藏书内容,开办法律讲座学习班,做好法律知识宣传工作,坚持以医教研为中心,深化服务质量。     

基层医院重症监护病房的医院感染特点及其对策

目的分析基层医院重症监护病房(ICU)的医院感染特点,提出相应的防范措施,以有效降低医院感染发生率. 方法以主动监测与系统回顾相结合的方法,对2003年全年入住ICU 383例患者的相关临床资料进行分析评判,并与全院住院患者进行对比. 结果 ICU的医院感染率是28.8%,显著高出同期的医院平均感染率23.6个百分点;因医院感染而导致死亡的占ICU总死亡的36%;ICU医院感染的病原菌以G-菌为主,占50%,而双重感染达78.7%;下呼吸道感染为医院感染的主要部位占61.7%,但多部位多器官感染占29.6%. 结论 ICU医院感染的特点是高发病率、高死亡率、高耐药性;其相关因素有病情重、年龄大、基础疾病多、侵入性诊疗操作多与交叉感染等.     

大鼠脊髓损伤修复差异表达cDNA文库的构建

目的　 运用改良消减杂交技术构建大鼠脊髓损伤修复过程中出现的差异表达基因文库 ,为寻找在脊髓损伤后的修复过程中起关键作用的分子提供帮助。方法　在经典消减杂交的基础上引进SMART反转录、长距离PCR、磁性分离系统、离心柱色谱等方法建立改良消减杂交技术 ,通过两轮消减杂交构建差异表达基因的cDNA消减文库 ,并对文库中的差异基因进行批量克隆与生物信息学分析。结果　 将两轮消减杂交后的差异表达基因转化大肠杆菌JM10 9以建立差异表达基因文库 ,库容量大小为 2× 10 3。从文库中随机挑取 90个克隆进行酶切鉴定与测序分析 ,得到 4 0个差异基因序列 ,其中 32个已知序列 ,8个新序列。 结论　 通过改良消减杂交成功建立了脊髓损伤修复过程中出现的差异表达基因文库 ,为筛选在大鼠脊髓损伤修复过程中起关键作用的基因 ,进一步探讨脊髓损伤修复的分子机制奠定了基础     

日本血吸虫磷酸丙糖异构酶编码基因的克隆与序列测定

目的克隆和鉴定日本血吸虫磷酸丙糖异构酶(SjTPI)编码基因,为寻找血吸虫病的候选疫苗联合应用打基础.方法设计合成引物,抽提日本血吸虫成虫总RNA,用RT-PCR法从中扩增出SjTPI基因编码序列,将其克隆入pGEM-T载体,用双酶切、以重组质粒为模板进行PCR扩增和测序进行鉴定.结果 RT-PCR法从成虫总RNA中扩增出大小为759 bp SjTPI基因编码序列,重组质粒pGEM-SjTPI经双酶切、PCR扩增,均可获得一条与RT-PCR产物一致的DNA片段,序列测定结果表明具有一个长度为759 bp的完整开放阅读框,与日本血吸虫(菲律宾株)和曼氏血吸虫磷酸丙糖异构酶核苷酸序列有高度同源性(分别为99%和88%).结论该实验成功地克隆了SjTPI编码基因,为进一步研究提供了条件.     

应用激光显微分离技术在骨肉瘤组织原位筛选噬菌体肽库

目的将激光显微分离(laser capture microdissection, LCM) 技术应用于噬菌体表面呈现肽库的筛选过程,建立一种可以直接在天然组织中筛选肽库的方法。方法将新鲜人骨肉瘤组织块在噬菌体肽库溶液振荡孵育后制成组织冰冻切片,免疫组化染色检测噬菌体在组织中的浸润扩散。改进常规LCM切片处理方法,以冻干法替代酒精/二甲苯法脱水,以期在LCM操作过程中提高切片上噬菌体的存活率。LCM法分离摄取骨肉瘤切片上的肿瘤靶细胞,转染回收这些细胞上特异结合的噬菌体。滴定法检测所筛选的噬菌体对特异性细胞的亲和力。结果利用LCM技术,可以由肿瘤冰冻切片上收集到足够的与特异性噬菌体短肽,应用于噬菌体表面呈现肽库筛选。经过3轮筛选后所获得的噬菌体与人骨肉瘤组织的特异性亲和力提高16倍。结论本研究首次将LCM技术应用于噬菌体表面呈现肽库的筛选,可以使我们直接在新鲜人肿瘤组织中筛选与特定细胞群甚至单个细胞亲合的短肽;同时又避免了天然组织中其他杂质细胞的污染,为研制细胞特异性导向载体提供了一个新的途径。     

各具特色的国外妇幼保健工作

本文通过对国际上几个主要国家的妇幼保健工作的介绍,使人们对不同特色的妇幼保健事业有所了解,他山之石,可以攻玉,希望能够对我国妇幼保健工作的发展提供借鉴。     

Comparison of the APACHE III, APACHE II and Glasgow Coma Scale in acute head injury for prediction of mortality and functional outcome

Objectives: This study examines the efficacy of the predicting power for hospital mortality and functional outcome of three different scoring systems for head injury in a neurosurgical intensive care unit (NICU). Design: On the day of admission, data were collected from each patient to compute the Acute Physiology, Age, and Chronic Health Evaluation (APACHE) II and III, and Glasgow Coma Scale (GCS) scores. Hospital mortality was defined as the deaths of patients before discharge from hospital. Early mortality was defined as death before the 14th day after admission. Late mortality was defined as death after the 15th day from admission. Functional outcome was evaluated by Index of Independence in Activities of Daily Living (Index of ADL). Setting: An 8-bed NICU in a 1270-bed medical center in Taichung Veterans General Hospital. Patients and participants: Two hundred non-selected patients with acute head injury were included in our study in a consecutive period of 2 years. Patients less than 14 years old were not included. Interventions: None. Measurements and results: Sensitivity, specificity and correct prediction outcome were measured by the chi-square method in three scoring systems. The Youden index was also obtained. The best cut-off point in each scoring system was determined by the Youden index. The difference in Youden index was calculated by Z score. A difference was also considered if the probability value was less than 0.05. The area under Receiver Operating Characteristic (ROC) curve was computed. Then the area under ROC of each scoring system was compared by Z score. There was statistical significance if p was less than 0.05. For prediction of hospital mortality, the best cut-off points are 55 for APACHE III, 17 for APACHE II and 5 for GCS. The correct prediction outcome is 82.4% in APACHE III, 78.4% in APACHE II and 81.9% in the GCS. The Youden index has best cut-off points at 0.68 for APACHE III, 0.59 for APACHE II, and 0.56 for GCS. The area under Receiver Operating Characteristic (ROC) curve is 0.90 in the APACHE III, 0.84 in the APACHE II and 0.86 in the GCS. There are no statistical differences among APACHE III and II, and GCS in terms of correct prediction outcome, Youden Index and the area under the ROC curve. Other physiological variables excluding GCS in APACHE III and II (AP III-GCS, AP II-GCS) have less statistical value in the determination of mortality for acute head injury. For the prediction of late mortality, APACHE III and II yield significantly better results in the area under the ROC curve, correct prediction and Youden index than those of GCS. Other physiological variables (AP III-GCS and AP II-GCS) play an important role in the prediction of late mortality in APACHE scores. For prediction of the functional outcome of surviving patients with acute head injury, the APACHE III yields the best results of correct prediction outcome, Youden index and the area under the ROC curve. Conclusion: The APACHE III and II may not replace the role of GCS in cases of acute head injury for hospital or early mortality assessment. But for prediction of the late mortality, the APACHE III and II have better accuracy than GCS. Other physiological variables excluding GCS in the APACHE system play a crucial contribution for late mortality. GCS is simple, less time-consuming and economical for patients with acute head injury for the prediction of hospital and early mortality. The APACHE III provides better prediction for severe morbidity than GCS and APACHE II. Therefore, the APACHE III provides a good assessment not only for hospital and late mortality, but also for functional outcome. Received: 22 May 1995 Accepted: 2 September 1996     

利用目的基因随机表位库筛选葡激酶抗原决定簇

目的：用目的基因随机表位库方法寻找葡激酶(straphylokinase SAK)的显性表位。方法：①SAK免疫BALB／C小鼠，免疫亲和层析纯化抗血清后，抗SAK抗体用生物素标记；②构建SAK随机表位肽库，随机挑取12个独立克隆测序，分析库DNA片段的分布和碱基含量情况；③以多抗为靶蛋白用克隆原位杂交法筛选肽库；④构建SAK的缺失突变体mSAK，Western blot分析mSAK的免疫反应性。结果：①筛选肽库得到一个由19个氨基酸组成的免疫显性表位区，命名为A1区；②mSAK不能与抗SAK多抗反应。结论：用简便、有效的方法筛选得到了SAK的一个表位A1区，初步确定A1区是SAK引起免疫反应的重要区域。