An Integrated Technique for Identification of Differential Genes Expressed in Patients with Cancer

Summary: To develop a method for identification of differential gene expression between different cell populations, several convenient techniques of molecular biology, including subtractive hybridization, suppression PCR, T/A cloning and sequencing, were used to identify genes expressed differentially in CD45^- and CD45^- cells isolated from U266 cell line of multiple myeloma. Our results showed that the levels of abundant genes scale down 20 times through subtractive hybridization.Plasmid DNA from CD45^- cell clones was hybridized with forward or backward cDNA probes synthesized from CD45^- and CD45 cells, respectively. A few of differentially expressed genes reconfirmed by RT-PCR were identified from 500 expressed clones of CD45^- cells. It is concluded that a strategy for gene expression identification developed from conventional molecular biological methods can be used in different laboratories.     

生物型人工硬脑膜应用的实验研究

OBJECTIVE: To evaluate the safety and efficacy of a dural graft prepared using porcine membrane in duraplasty. METHODS: Eighteen New Zealand rabbits were randomly divided into groups A (n=4), B (n=4), C (n=5), and D (n=5) sacrificed 3, 14, 30 and 90 d after duraplasty, respectively. Each animal underwent bilateral parietal craniectomy behind the coronal suture and beside the midline to expose the dura, which was cut on the right side and substituted with the dural graft. The exposed dura on the left was kept intact as control. The rabbits were observed for WBC counts before the operation and before sacrifice by transcardiac formalin perfusion, respectively. The meninges and brain tissues were histologically examined after sacrifice. RESULTS: The WBC count varied little after the operation (P>0.05). Microscopic examination demonstrated tissue repair on both the implantation side and control side, without graft adhesion to the cortical surface. In group A, a large number of leukocytes were seen gathering on the lateral dura, suggesting acute tissue repair. In group B, endothelial cells covering the inner surface of the graft could be seen. Fibroblasts and fibrocytes were seen in the grafts between collagen fibers in group C, and in group D, fibroblasts and fibrocytes increased between the collagen fibers and the suture healed. CONCLUSION: The dura graft is safe and applicable for dural defect repair.     

急性重型颅脑损伤ET和CGRP含量变化及其临床意义

目的　探讨急性重型颅脑损伤 (ASBI)患者血浆中内皮素 (ET)和降钙素基因相关肽 (CGRP)的含量变化及其临床意义。方法　用放免法测定 2 8例ASBI患者伤后 6～ 2 4h血浆中ET及CGRP含量。结果　ASBI患者ET含量明显增多 ,差别有显著性 (P <0 .0 5 ) ;CGRP也有一定的增多 ,但无显著性差异。结论　ET和CGRP共同参与ASBI病理生理反应过程 ,ET释放增加是加重脑继发性损害的重要因素     

黄芪注射液对糖尿病肾病患者尿Ⅳ型胶原的影响及意义

目的 观察黄芪注射液对糖尿病肾病（DN）患者尿Ⅳ型胶原(CⅣ)探讨黄芪注射液治疗DN的机制。方法 分别测定黄芪注射液治疗组(30例)，对照组(28例)治疗前后尿CⅣ、24小时尿白蛋白排泄率(UAE)、α1-微球蛋白(α1-MG）。结果 黄芪注射液能明显降低DN患者尿CⅣ水平，减少DN患者UAE、α1—MC。结论 黄芪注射液对DN患者肾脏有保护作用，可能与其抑制DN患者CⅣ有关。     

Ⅰ、Ⅲ型前胶原基因第2外显子核酶对靶RNA的体外切割活性的研究

目的 体外研究锤头型α1Ⅰ型及Ⅲ型前胶原基因第2外显子片段核酶对各自靶RNA分子的切割活性及反应条件。同时观察两反义核酶对瘢痕中成纤维细胞胶原合成的影响。方法 将含α1Ⅰ型及Ⅲ型前胶原基因第2外显子片段的重组质粒(pT-Ⅰ、pT-Ⅲ)，经体外^32P标记转录后形成产物靶RNA。同时将含特异性核酶基因的重组质粒(pT-gⅠ、pT-gⅢ)进行非标记的体外转录，产物(核酶)与各自的^32P-靶RNA按不同的条件混和反应，电泳后放射自显影观察结果。将构建好的核酶以脂质体包裹后导入培养的成纤维细胞内，采用图像分析法观察核酶对成纤维细胞Ⅰ型、Ⅲ型胶原蛋白mRNA合成的影响。结果 两种核酶在37℃、42℃均能有效切割各自的靶RNA，对Mg^2 浓度的要求范围较宽(10～20mmol／L)；反应温度从65℃逐渐降至并维持在37℃的条件下核酶切割活性显提高。Ⅰ、Ⅲ型胶原蛋白mRNA的表达明显降低，胶原蛋白生成降低，胶原生成明显受抑制。结论 针对α1Ⅰ型及Ⅲ型前胶原基因第2外显子片段的核酶能有效地在体外对靶RNA进行切割并能有效地抑制瘢痕中成纤维细胞胶原的合成。     

星状神经节阻滞治疗交感型颈椎病12例分析

交感型颈椎病临床主要表现为疼痛、感觉异常和植物神经功能紊乱,目前以非手术治疗为主,方法较多[1～3],各具特色.我科自2002年10月以来采用星状神经节阻滞治疗交感型颈椎病12例,取得满意的效果,总结如下.     

胆囊胆固醇结石病的遗传基础

胆囊胆固醇结石病的发生与胆汁胆固醇过饱和、成核缺陷以及胆囊收缩功能异常有关，同时受到遗传因素的影响。为进一步探索胆石病发病机制，复习近年来国内外胆石病的遗传基础研究取得的进展。     

凝血因子VII及其基因多态性与冠心病

大部分的冠脉缺血性事件 (包括猝死、心肌梗死和不稳定型心绞痛 )是因粥样斑块破裂后触发血栓形成所致。凝血功能失调在冠心病的发病中起重要作用 ,尤其在外源性凝血途径中发挥主要作用 ,凝血因子VII(FVII)是该途径中起关键作用的因子     

颅脑损伤患者脑型肌酸激酶同工酶变化的临床意义

近年研究发现，颅脑损伤患者脑型肌酸激酶同工酶活性(CK-BB)均有不同程度的升高。2001年11月至2002年9月，我们对52例急性颅脑损伤患者进行血清CK-BB活性测定，旨在探讨急性颅脑损伤患者CK-BB活性变化的临床意义。