RNA干扰基因敲除MAGE-1在恶性胶质瘤U87细胞中的初步研究

目的：通过构建抑制MAGE-1的短片段双链核糖核酸（siRNA）表达载体，鉴定其在人恶性胶质瘤细胞系U87细胞中对MAGE．1基因表达的干涉作用。方法：化学合成2对编码短发夹RNA序列的靶向MAGE—1基因寡核苷酸链，克隆至经BglⅡ、HindⅢ双酶切的pSUPER载体上，重组构建核糖核酸干扰（RNAi）质粒载体。利用RT-PCR、流式细胞术和荧光显微镜，检测经稳定转染后胶质瘤U87细胞中MAGE-1的表达，以了解siRNA的干涉效果。结果：重组构建的pSUPER—MAGE—1载体经双酶切、电泳及插入基因片段序列分析，表明寡核苷酸链成功地插入至预计位点，且序列与预期完全一致。稳定转染后G418筛选出的U87多克隆细胞MAGE-1的表达经RT—PCR、流式细胞术和荧光显微镜检测，2对siRNA均有较明显的干扰作用。结论：载体的成功构建并能对U87细胞中的MAGE—1分子进行RNAi，为进一步研究MAGE—1在肿瘤中的作用，分析基因功能，展开肿瘤基因治疗奠定了基础。     

门诊护理投诉原因分析及对策

目的分析门诊常见护理投诉原因并提出护理对策。方法采用回顾性研究分析,统计我院近3年门诊护理投诉,并对其进行调查核实,详细分析原因。结果近3年门诊护理投诉共78例,其中有效护理投诉42例（53.85%）;不同职称、工龄护理人员被投诉率比较,差异有显著性意义（均P〈0.01）。结论服务意识淡薄、护患沟通不良是门诊护理投诉的主要原因,通过转变服务意识,再造流程,增加便民服务等措施能及时化解护患矛盾,提高患者的满意度。     

大咯血的急诊外科治疗

目的　总结 4 4a急诊手术治疗大咯血经验。方法　回顾性研究 195 8～ 2 0 0 1年在本院急诊手术的 89例大咯血患者 ,收集临床表现、诊断方法、原发病因、治疗和结果等相关资料。结果　 85 4 %病例的X线表现与出血部位一致 ,其它诊断方法 (支气管镜、CT、血管造影术等 )作为重要补充。由于 1982年后的早期外科干预 ,术前咯血总量、咯血平均量和低血压、休克发生比例较前明显减少 ,术后并发症从 1982年前的 2 2 2 %降至 17 1% ,围手术期死亡率从 9 3%降至 0。急诊全肺切除术并发症 ( 2 3 3% )高于肺叶切除术 ( 15 2 % )。在过去 4 4a肺结核一直是大咯血的主要原发病因。结论　符合新的外科治疗标准的患者应及早手术 ;选择切除范围要保守 ,首选叶切     

良性前列腺增生中医证型与尿动力学参数相关性研究

目的:探讨良性前列腺增生(BPH)中医证型与尿动力学参数的相关性。方法:152例BPH患者,中医辨证分为肾阴不足、肾阳虚弱、瘀阻水道、脾气虚弱、肺热气郁、湿热下注、痰浊郁结七型,均进行尿动力学检查。结果:152例BPH患者中,肾阳虚弱型71例(46.71%),瘀阻水道型40例(26.31%),肾阴不足型14例(9.21%)。膀胱出口梗阻为III～VI度肾阳虚弱型有58例,瘀阻水道型有38例,其中严重梗阻(V～VI)26例。膀胱逼尿肌收缩功能极弱中(n=12),肾阳虚弱型4例,占33.33%(4/12),瘀阻水道型7例,占58.33%(7/12);膀胱逼尿肌收缩功能弱者中(n=48),肾阳虚弱型为27例,占56.25%(27/48),瘀阻水道型17例,占35.42%(17/48)。结论:BPH中医各辨证分型与膀胱出口梗阻和膀胱逼尿肌收缩力有一定的相关性,从而为中医证型的量化和客观化提供相应的证据。     

脂肪抽吸术对胰岛素敏感性及脂质代谢的影响

目的了解脂肪抽吸术对胰岛素抵抗与脂质代谢的影响,并探讨其临床意义。方法对20例接受腹部脂肪抽吸术者术前及术后2个月的血清三酰甘油、总胆固醇、高密度脂蛋白胆固醇、低密度脂蛋白胆固醇,胰岛素敏感性进行测量并进行比较。结果术后与脂肪抽吸术前比较,术后胰岛素敏感性增强（P〈0.01）,血清总胆固醇与低密度脂蛋白胆固醇水平下降。结论腹部脂肪抽吸术在去除大量皮下脂肪的同时,对胰岛素抵抗、脂质代谢等心脑血管危险因素也有非常有益的影响。     

^1H-magnetic resonance spectroscopy screening for animals with acute cerebral infraction suitable forthrombolytic therapy

BACKGROUND: As a non-invasive technique which can provide comprehensive biological information, 1H-magnetic resonance spectroscopy (1H-MRS) may provide valuable reference data for irreversible recovery or reversible changes in ischemic tissue after stroke. OBJECTIVE: To monitor and evaluate the effect of the urokinase thrombolytic therapy after experimental acute cerebral ischemia by 1H-MRS technology and investigate its adaptability. DESIGN: Randomly controlled animal study. SETTINGS: Shenzhen Hospital of Peking University and National Key Laboratory of Pattern and Atom & Molecular Physics, Wuhan Physics and Mathematics Institute, Chinese Academy of Science. MATERIALS: Eleven healthy adult Sprague-Dawley (SD) rats, weighing 260–300 g and of both genders, were supplied by Experimental Animal Center of Tongji Medical Collage, Huazhong University of Science and Technology [SCXK (e) 2004-007]. 4.7T superconducting nuclear magnetic resonance meter was provided by Brucker Company. METHODS: The experiment was carried out in Shenzhen Hospital of Peking University and National Key Laboratory of Pattern and Atom & Molecular Physics, Wuhan Physics and Mathematics Institute, Chinese Academy of Science from August 2003 to December 2005. ① The rats were randomly divided into 30-minute self-thrombo-embolism group (n =6) and 60-minute self-thrombo-embolism group (n =5). Six rats in 30-minute self-thrombo-embolism group were occluded with clot embolus for 30 minutes and 5 rats in 60-minute self-thrombo-embolism group were occluded for 60 minutes. 10 000 U/kg urokinase was dissolved in 2 mL saline and the operation lasted for 5 minutes. ② 1H-MRS was performed before thrombolysis and at 3 hours and 24 hours after successful embolization. The metabolic changes of N-acetyl-L-aspartic acid (NAA)/phosphocreatine (PCr) + creatine (Cr), choline phosphate (Cho)/PCr+Cr and lactic acid (Lac)/PCr+Cr in the region of interests were analyzed. ③ The T2W image was conducted 24 hours after the thrombolytic therapy with TR=500 ms and TE=25 ms. ④ The subjects were sacrificed immediately after 1H-MRS and the brain tissues were cut into pieces and stained with HE method; in addition, pathological changes were observed under optic microscope. MAIN OUTCOME MEASURES: ① Metabolic changes of NAA/PCr+Cr, Cho/PCr+Cr and Lac/PCr+Cr in the region of interests; ② T2W image at 24 hours after the thrombolysis; ③ pathological observation of brain tissue. RESULTS: Eleven rats were all involved in the final analysis. ① Metabolic changes in the region of interests : In 30-minute self-thrombo-embolism group, the Lac peak emerged immediately after the embolism, but the ischemic zone decreased 3 hours after the thrombolytic therapy (0.252±0.01, 0.603±0.01, P < 0.01). Lac/(PCr+Cr) ratio was 0.290±0.01 at 24 hours after thrombolysis, which was higher than that at 3 hours after thrombolysis (P < 0.01). The NAA/ (PCr+Cr) ratio decreased significantly at 3 hours after the thrombolysis as compared with that before thrombolysis (0.922±0.16, 1.196±0.01, P < 0.05). In 60-minute self-thrombo-embolism group, the Lac/(PCr+Cr) ratio was higher at 3 hours after thrombolysis than that before thrombolysis (0.846±0.12, 0.601±0.11, P < 0.05) and the NAA/(PCr+Cr) decreased at 3 hours after the embolism. Fluctuation of NAA/ (PCr+Cr) ranged from 0.68 to 0.75 before thrombolysis and from 0.71 to 0.75 at 3 hours after thrombolysis. ② T2W image: T2W image showed that 2 subjects in 30-minute self-thrombo-embolism group whose Lac/NAA was higher than 0.7 suffered from intracranial hemorrhage. This meant that the subjects with Lac/NAA > 0.7 were more likely to suffer from intracranial hemorrhage. ③ Histological and morphological examinations: Optic microscope demonstrated that interspace surrounding nerve cells was widened at ischemic center; neurons were swelling; nucleus was stained lightly; pyknosis and mesenchymal edema were mainly observed in lateral cortex of brow and vertex and in lateral part of corpus striatum. CONCLUSION: ①Compound parameters in ischemic area before thrombolysis should be regarded as an important predicting marker for thrombolytic therapy, effect evaluation and termination. ② 1H-MRS combining with other imaging technique is a detecting way for screening cases who are suitable for thrombolytic therapy.     

与甲壳素缝线体外复合培养大鼠成肌细胞株L6细胞的形态学

目的：探讨甲壳素作为骨骼肌组织工程支架材料的可行性。 方法：实验于2002-06／12在南方医科大学应用解剖学研究所完成。大鼠成肌细胞株L6与甲壳素医用缝合线体外复合培养。倒置相差显微镜下观察细胞的形态和排列情况。并分别在体外复合培养的第1，2，3，4，5，6天，取出细胞支架复合体，扫描电镜观察成肌细胞的形态和排列情况。 结果：①扫描电子显微镜观察显示，甲壳素医用可吸收缝线的表面比较粗糙。②倒置相差显微镜下可见大鼠L6细胞与甲壳素制备的医用可吸收缝合线间黏附良好，细胞沿缝线排列呈现串珠样，并可以见到细胞聚集现象。③扫描电子显微镜观察，接种24h后，细胞主要为圆球形，随后细胞迁移、伸展、并见突起；第3天，成肌细胞主要为梭形，并见融合趋势；4d后，细胞沿材料纵轴排列并相互融合，并见肌小管样结构形成及细胞外基质分泌。 结论：甲壳素具有作为支架以构建组织工程化骨骼肌的潜能，支架的平行排列有助于工程化骨骼肌纤维良好方向性的形成。     

547例垂体腺瘤延迟诊断原因及影像诊断分析

目的:提高垂体腺瘤特别是微腺瘤早期影像诊断率。方法:通过对所搜集病史资料完整,经医学影像学确诊和手术病理证实的547例垂体腺瘤(内有251例微腺瘤)延迟诊断原因的分析,找到影响垂体腺瘤早期诊断的因素。结果:296例垂体大腺瘤蝶鞍X线平片、CT、MR常规检查即可诊断。CT骨窗蝶鞍片有助于垂体微腺瘤的诊断,251例微腺瘤经高分辨力CT薄层冠状强化扫描和高场强MR薄层强化扫描得以明确诊断。结论:熟悉垂体腺瘤的首发症状,及时行CT、MR常规扫描可明确大腺瘤的诊断,CT骨窗片可发现垂体微腺瘤的佐证。高场强MR薄层强化扫描是目前诊断垂体微腺瘤的最佳方法,高分辨力CT冠状薄层强化扫描次之,结合神经内分泌学检查是提高垂体腺瘤影像学早期诊断率的主要措施。     

斑马鱼：肾脏发育与疾病研究的理想模式生物

斑马鱼（zebrafish）为小型热带鱼类，具有世代周期短（2～3个月）、产卵量大、形体较小（成体仅长3～4cm）的特点，尤其是卵子体外受精，胚胎发育速度快，仅需2．5d就可发育为幼鱼。此外，斑马鱼的精子还可通过冷冻来保存，而且胚体完全透明，胚胎发育过程可以直接观察使得发育异常的突变体很容易被识别，这就给遗传操作和人工诱变提供了极为有利的条件。