老年同型半胱氨酸与C反应蛋白水平与冠心病发病的相关性

目的观察老年冠心病（CHD）患者血浆同型半胱氨酸（Hcy）、C反应蛋白（CLIP）、低密度脂蛋白胆固醇（LDL-C）水平的变化及临床意义。方法检测并比较42例稳定型心绞痛患者（SAP组）、67例急性冠状动脉综合征患者（ACS组）及82例健康查体者（对照组）的Hcy、ClIP、LDL-C水平。结果SAP、ACS组血浆Hcy、CRP、LDL-C水平明显高于对照组（P均〈0．05）,SAP与ACS组比较,Hcy、CRP水平有统计学差异（P〈0．01）,LDL-C水平无统计学差异（P〉0．05）;不同狭窄支数患者的Hcy、CRP水平比较有统计学差异（P〈0．05或〈0．01）。结论Hcy和CRP可作为预测老年CHD严重程度的指标,LDL-C对老年CHD严重程度的预测价值不高。     

Expression and Fuactional Role of HERG1, K~+ Channels in Leukemic Cells and Leukemic Stem Cells

In order to investigate the expression and functional role of HERG1 K channels in leukemic cells and leukemic stem cells (LSCs), RT-PCR was used to detect the HERG1 K channels expression in leukemic cells and LSCs. The functional role of HERG1 K channels in leukemic cell proliferation was measured by MTT assay, and cell cycle and apoptosis were analyzed by flow cy- tometry. The results showed that herg mRNA was expressed in CD34 /CD38-, CD123 LSCs but not in circulating CD34 cells. Herg mRNA was also up-regulated in leukemia cell lines K562 and HL60 as well as almost all the primary leukemic cells while not in normal peripheral blood mononuclear cells (PBMNCs) and the expression of herg mRNA was not associated with the clinical and cytoge- netic features of leukemia. In addition, leukemic cell proliferation was dramatically inhibited by HERG K channel special inhibitor E-4031. Moreover, E-4031 suppressed the cell growth by induc- ing a specific block at the G1/S transition phase of the cell cycle but had no effect on apoptosis in leukemic cells. The results suggested that HERG1 K channels could regulate leukemic cells prolif- eration and were necessary for leukemic cells to proceed with the cell cycle. HERG1 K channels may also have oncogenic potential and may be a biomarker for diagnosis of leukemia and a novel potential pharmacological target for leukemia therapy.     

Expression and fuactional role of HERG1, K<Superscript>+</Superscript> channels in leukemic cells and leukemic stem cells

Summary In order to investigate the expression and functional role of HERG1 K⁺ channels in leukemic cells and leukemic stem cells (LSCs), RT-PCR was used to detect the HERG1 K⁻ channels expression in leukemic cells and LSCs. The functional role of HERG1 K⁺ channels in leukemic cell proliferation was measured by MTT assay, and cell cycle and apoptosis were analyzed by flow cytometry. The results showed that herg mRNA was expressed in CD34⁺/CD38⁻, CD123⁻ LSCs but not in circulating CD34⁺ cells. Herg mRNA was also up-regulated in leukemia cell lines K562 and HL60 as well as almost all the primary leukemic cells while not in normal peripheral blood mononuclear cells (PBMNCs) and the expression of herg mRNA was not associated with the clinical and cytogenetic features of leukemia. In addition, leukemic cell proliferation was dramatically inhibited by HERG K⁺ channel special inhibitor E-4031. Moreover, E-4031 suppressed the cell growth by inducing a specific block at the G1/S transition phase of the cell cycle but had no effect on apoptosis in leukemic cells. The results suggested that HERG1 K⁺ channels could regulate leukemic cells proliferation and were necessary for leukemic cells to proceed with the cell cycle. HERG1 K⁺ channels may also have oncogenic potential and may be a biomarker for diagnosis of leukemia and a novel potential pharmacological target for leukemia therapy. LI Huiyu, female, born in 1960, Associate Professor This project was supported by a grant from National Science Foundation for Distinguished Young Scholars of China (No. 30225038).     

WISp39基因对U937细胞增殖、凋亡和周期的影响

为了探讨一种新发现的p21调控蛋白WISp39对白血病细胞的增殖、凋亡和周期的影响,构建了WISp39的真核表达质粒pLenti6/V5-WISp39,并导入了人髓系白血病细胞系U937细胞,转染后用定量PCR检测了WISp39表达的变化,用CCK-8法测定细胞增殖活性,流式细胞术（FACS）检测细胞凋亡和周期的变化。结果显示：pLen-ti6/V5-WISp39转染48小时后,实验组中WISp39mRNA表达上升了（5.5±1.2）倍,同时U937细胞的增殖明显受到抑制,抑制率为37.6%;进一步的分析表明,在实验时间内,WISp39表达的升高不能明显诱导U937细胞的凋亡,但G0/G1期细胞比例由（40.59±0.7）%上升到（49.79±1.1）%。结论：WISp39对于U937无明显的诱导凋亡作用,但通过对细胞周期的影响,使停滞在G/G期的细胞增多,从而抑制了U937增殖。     

SARI基因过表达对K562细胞系生物学影响的研究

目的:通过构建SARI慢病毒表达载体,探讨SARI基因过表达对K562细胞生物学功能的影响。方法:采用RT-PCR方法获得目的基因并重组p LOV.CMV.GFP质粒,构建pLOV.CMV.GFP-SARI表达载体。通过测序鉴定、病毒包装和滴度测定后,获得阳性病毒颗粒,并以病毒颗粒感染K562细胞系。采用Q-PCR及Western blot方法验证感染后K562细胞SARI基因转录和蛋白水平的表达情况,同时采用CCK-8法检测K562细胞的增殖情况,以流式细胞术分析细胞凋亡和细胞周期变化。结果:利用RT-PCR方法获得SARI基因并成功构建了含SARI基因的慢病毒表达载体,从分子及蛋白水平验证了其在感染后K562细胞中的高表达;与空白组和感染空载体的Mock组相比,过表达SARI载体组的细胞增殖显著受抑、细胞凋亡率增加,而细胞周期无显著变化。结论:SARI基因过表达可抑制K562细胞的增殖并促进其凋亡,提示诱导SARI基因过表达可能对于抑制CML发生发展具有重要的作用。