结核分枝杆菌Rv1009基因蛋白的克隆和表达

目的 克隆表达结核分枝杆菌促Rv1009基因,序列测定正确后进行融合、表达.方法 采用热启动聚合酶链反应（Polymerase Chain Reaction,PCR）从结核分枝杆菌H37Rv基因组中扩增出Rv1009编码基因,用限制性内切酶消化后插入pGEX 4T-2载体中,将重组质粒转化大肠杆菌BL21（DE3）,目的基因经IPTG诱导,表达Rv1009基因蛋白.结果 经PCR扩增在1300bp处发现一条目的片段,获得了结核分枝杆菌H37Rv株Rv1009基因蛋白,经诱导后高效表达分子量为64KD的外源蛋白,与预期分子量大小一致,凝胶自动扫描分析,在A600值为0.6,IPTG终浓度为0.3 mmol/L,诱导表达3 h时融合蛋白表达量即达峰值,占菌体总蛋白的22.8%.结论 构建了结核分枝杆菌Rv1009基因重组表达载体,获得了RPF样融合蛋白的高效表达,为今后深入研究奠定了基础.     

Stereoselective metabolism of lansoprazole by human liver cytochrome P450 enzymes.

The stereoselective metabolism of lansoprazole enantiomers was evaluated by incubation of human liver microsomes and cDNA-expressed cytochrome p450 (p450) enzymes to understand and predict their stereoselective disposition in humans in vivo. The intrinsic clearances (Clint) of the formation of both hydroxy and sulfone metabolites from S-lansoprazole were 4.9- and 2.4-fold higher than those from the R-form, respectively. The sums of formation Clint of both metabolites were 13.5 and 57.3 microl/min/mg protein for R- and S-lansoprazole, respectively, suggesting that S-lansoprazole would be cleared more rapidly than the R-form. The p450 isoform selective inhibition study in liver microsomes, and the incubation study of cDNA-expressed enzymes, demonstrated that the stereoselective sulfoxidation is mediated by CYP3A4 and that the hydroxylation is mediated by CYP2C9 and CYP3A4 as well as by CYP2C19. Total Clint values of hydroxy and sulfone metabolite formation catalyzed by all these p450 enzymes were consistently higher for S-lansoprazole than for the R-form. The CYP3A4 produced the greatest difference of Clint between S- and R-enantiomers, mainly due to a difference of sulfoxidation metabolism (Clint 76.5 versus 10.8 microl/min/nmol of p450, respectively), whereas CYP2C19-catalyzed hydroxylation resulted in a minor difference of Clint between S- and R-enantiomers (179.6 versus 143.3 microl/min/nmol of p450, respectively). However, the affinity of CYP2C19 on hydroxylation was 5.7-fold higher for S-enantiomer than for the R-form (Km 2.3 versus 13.1 microM), suggesting that the role of CYP2C19 on stereoselective hydroxylation would be more prominent at concentrations around the usual therapeutic level. These findings suggest that both CYP2C19 and CYP3A4 are major enzymes contributing to the stereoselective disposition of lansoprazole, but stereoselective hydroxylation of lansoprazole enantiomers is mainly influenced by CYP2C19, especially at the usual therapeutic doses.     

中风后癫痫的临床研究

本文报告３６例中风后癫痫发作的临床和ＣＴ资料。通过回顾性病历复习和随访发现，中风后癫痫的发生率占同期住院中风患者的５．２６％，以蛛网膜下腔出血和脑栓塞发生癫痫比率最高，分别为１５％和１２．５％。癫痫发作与ＣＴ所见病灶分布密切相关，皮质病灶较皮质下病灶更易发生病病。癫痫发作可发生于中风后任何时期，但早期癫痫以出血性中风多见，而迟发性癫痫则更多见于脑梗塞患者。     

33例肝占位性病变MRI误诊的分析

我院１９８８年以来４０００余例腹部ＭＲＩ中，发现肝占位性病变误诊３３例。所用机型为美国Ｄｉｓｏｎｉｃｓ公司０．５Ｔ超导ＭＲＩ，自旋回波序列（ＳＥ序列）、常规Ｔ１加权（Ｔ１ＷＩ）、质子加权（ＰＤＷＩ）、Ｔ２加权（Ｔ２ＷＩ），覆盖全肝。其中，ＰＨＣ误诊为ＭＨＣ５例、ＭＨＣ误诊为ＰＨＣ６例、ＰＨＣ误为ＨＨＥ５例、ＭＨＣ误诊为ＨＨＥ２例、ＨＨＥ误诊为ＰＨＣ６例、肝硬化结节误诊为ＰＨＣ２例、炎性假瘤误诊为ＰＨＣ３例、肝结核误诊为ＰＨＣ１例、ＨＣＹ误诊为ＨＨＥ３例。本文从病变的影像学特征和扫描技术方面详细探讨了误诊的原因和鉴别诊断要点。     

Thymidine labeling index in breast tumors

The relationship between primary tumor proliferative activity and clinical and pathologic characteristics was analyzed in relation to menopausal status in 32 patients with malignant or benign breast disease. The thymidine labeling index (TLI) showed significantly higher median values in the cancer patients (3.48 per cent) than in the patients with benign diseases (1.02 per cent). TLI was not significantly affected by delayed incubation at room temperature for about 1 hour. In the breast cancer patients, TLI did not significantly correlate to tumor size, the presence of axillary lymph node metastasis or pathologic nuclear grading. The only significant difference was limited to the breast cancer patients without axillary lymph node metastasis in relation to menopausal status; the TLI in the premenopausal patients (5.10 per cent) was significantly higher (p<0.05) than that in the postmenopausal patients (2.28 per cent). These data thus suggest that among premenopausal patients without axillary lymph node metastasis, those with a high TLI could be potential candidates for adjuvant chemotherapy.     

通塞益脑液治疗急性脑梗塞的机理探讨

本文以结扎左侧颈总动脉造成实验性急性脑梗塞的沙土鼠模型,探讨通塞益脑液治疗急性脑梗塞的机理。发现结扎术前胃饲通塞益脑液者,可降低急性脑梗塞后脑组织中过氧化脂质的含量;结扎术后给药者,可降低脑组织中钙的含量。初步认为通塞益脑液对急性脑梗塞的脑组织有保护作用。     

新型麻醉药异丙酚的氧化动力学

采用高效液相色谱法测定了异丙酚在不同温度下及其甲醇 水（Ｖ∶Ｖ＝７０∶３０）混合溶液在不同ｐＨ值下的反应速率常数，并求出了异丙酚氧化反应的活化能，结果表明ｐＨ值与温度对异丙酚的氧化动力学均有影响     

Expression and distribution of vascular endothelial growth factor protein in human brain tumors

Marked neovascularization is a hallmark of many neoplasms in the nervous system. Recent reports indicate that the endothelial mitogen vascular endothelial growth factor (VEGF) may play a critical role in the regulation of vascular endothelial proliferation in malignant gliomas. Using novel monoclonal antibodies to the VEGF polypeptide we have determined the expression and cellular distribution of VEGF protein in a representative series of 171 human central nervous system (CNS) tumors by immunohistochemistry and immunoblotting. In agreement with previous in situ hybridization data, 19 out of 20 glioblastomas (95%) showed immunoreactivity for VEGF, whereas both the percentage of immunoreactive tumors and the extent of immunoreactivity for VEGF were significantly lower in astrocytomas. Of the pilocytic astrocytomas (WHO grade I) 44% were immunoreactive for VEGF, but we observed several cases with pronounced vascular proliferates in the absence of VEGF. In ependymomas, meningiomas, hemangioblastomas, and primitive neuroectodermal tumors, there was no correlation between VEGF expression, vascular endothelial proliferation and the grade of malignancy. Oligodendrogliomas and the oligodendroglial component of mixed gliomas lacked immunoreactive VEGF, indicating that endothelial growth factors other than VEGF may regulate tumor angiogenesis in these neoplasms. Western blot analysis showed a predominant VEGF protein species of 23 kDa and confirmed the immunohistochemical data in all cases. Our findings demonstrate that VEGF is expressed in a wide spectrum of brain tumors in which it may induce neovascularization. However, other angiogenic factors also appear to contribute to the vascularization of CNS neoplasms. Received: 18 April 1996 / Revised, accepted: 20 August 1996     

二维彩色多普勒超声心动图诊断扩张型心肌病（附100例分析）

应用ＡＴＬ－ｕｌｔｒａｍａｒｋ超６型彩色多普勒超声诊断仪诊断扩张型心肌病１００例，并与性别、年龄相应的正常人对比分析。结果：除扩张型心肌病各组室间隔及左室后壁舒张期厚度与相应正常对照组相比无显著性差异（Ｐ＞０．０５）外，其左室舒张内径及收缩内径、左房内径明显大于正常对照组（Ｐ＜０．０１），左室射血分数及左室内径缩短分数较正常对照组明显降低（Ｐ＜０．０１），并见心腔内血流速度缓慢，颜色变淡。另在７７例病人中探及４０例瓣膜返流。提示该诊断方法对扩张型心肌病具有直观、简便、准确、迅速的优点，为本病的诊断及鉴别诊断提供了良好的手段。