三七皂苷单体对高血压病人血小板功能的影响

目的:以硝苯地平及1{β[3-4-甲基苯丙氧基-甲氧苯]-1-H-盐酸咪唑}(SK&F96365)为对照,探讨三七皂苷单体人参皂苷-2A对血小板聚集功能的作用。方法:肘静脉采高血压病人血标本30份,正常人血标本30份,常规制备富含血小板血浆,分别用不同浓度的硝苯地平、SK&F96365、人参皂苷-2A孵育,以2μmol/L二磷腺苷为诱聚药,连续5次测定5分钟血小板的聚集率,以血小板最大聚集率(maximal platelet aggregation,PAGmax)为观察指标。结果:①高血压病人PAGmax为0.89±0.06,与正常人PAGmax0.68±0.07比较,差异有统计学意义(P<0.01);②两种浓度(10 μmol/L、20 μmol/L)的硝笨地平时正常人和高血压病人PAGmax均无影响;③4种浓度(2.5 μmol/L、5μmol/L、10μmol/L、20μmol/L)的SK&F96365对高血压病组的PAGmax均有抑制作用(均为P<0.05);④4种浓度(2.5μmol/L、5μmol/L、10μ,mol/L、20 μmol/L)的人参皂苷-2A对高血压病组的PAGmax均有抑制作用(均为P<0.05);3种浓度(5 μmol/L、10 μmol/L、20 μmol/L)的人参皂苷-2A对正常组的PAGmax均有抑制作用(均为P<0.05)。结论:高血压病人血小板聚集功能明显高于正常人,血小板呈高反应状态,是血栓形成的重要因素之一;硝苯地平不能抑制血小板聚集;SK&F96365能抑制血小板的聚集功能;人参皂     

家兔动脉粥样硬化与血小板源内皮型氧化氮合酶表达的相关性

Objective To explore the correlation of atheroselerosis progression and the expression of platelet derived endothelial nitric oxide synthase (eNOS) in rabbits. Methods A total of 24 male New Zealand white rabbits were used in this study. Six of the animals were fed with normal food (control group). Eighteen rabbits were fed with cholesterol-rich food (1 g/d) for 12 weeks to establish the atherosclerosis model. Among 18 models, 6 rabbits were executed immediately and their aorta and platelet samples were collected for further analysis (model group), 6 rabbits were orally administered with pravastatin (10 rag/d) for additional 12 weeks (treated group), and the remaining 6 rabbits were left untreated until the end of the study (untreated group). The control, treated and untreated animals were then killed, and the aorta and platelet samples were collected for eNOS expression analysis (RT-PCR). Results The aorta samples in model and untreated group exhibited rough intima and a lot of longitudinal fatty streaks, which indicated that atherosclerosis models were established successfully. While in treated group, the degree of atherosclerosis was decreased. The average percent of thickness of fatty streaks or atheroselerotic plaques relative to the whole thickness of vessel walls was 0. 04±0. 02, 0. 82±0. 16, 0. 33±0. 18,0. 77±0. 14 in control, model, treated and untreated group, respectively. The thickness of fatty streaks or atherosclerotic plaques was significantly increased in the model and untreated groups and decreased in treated group compared with the control group (both P<0. 05). The expressions of platelet derived eNOS/mRNA were 1. 02± 0. 28, 0. 41± 0. 27, 1.00 ± 0. 77, 0. 40±0. 29 in control, model, treated and untreated group, respectively. The expression of eNOS/mRNA was markedly decreased in model group and untreated group compared with the control group, but was increased in treated group compared with untreated and model groups (F=3. 544, P = 0. 024). Conclusions There is a negative correlation between eNOS expression and atherosclerosis development, which suggests that the reversal effect of pravastatin on atheroselerosis progression and plaque formation may relate to the expression of platelet derived eNOS.     

获得性血友病甲2例报告并文献复习

目的: 探讨获得性血友病甲的临床特点,以提高对该病的认识.方法: 分析2例获得性血友病甲患者的临床资料,包括其临床表现、实验室检查及治疗经过,并结合相关文献进行复习.结果与结论: 2例患者均为老年男性,表现为自发性广泛皮下出血及软组织血肿,活化部分凝血活酶时间(activated partial thromboplastin time,APTT)明显延长且不能被正常血浆纠正,血浆凝血因子Ⅷ凝血活性部分(factor Ⅷ coagulant activity,FⅧ :C)活性下降,抗FⅧ:C抗体滴度升高,予输注FⅧ浓缩制剂及肾上腺皮质激素、人血丙种球蛋白后出血控制.获得性血友病甲可致严重出血,及早诊断、及早控制出血、及时补充缺乏的凝血因子,清除抗FⅧ:C抗体是成功治疗的关键.     

特发性血小板减少性紫癜242例临床分析

目的:总结儿童和成人特发性血小板减少性紫癜(idiopathicthrombocytopenicpurpura,ITP)患者的临床特点。方法:对242例ITP患者的临床资料进行回顾性分析。结果:儿童ITP组,140例,男女比例为1.5∶1,急性型87例(62.1%),起病前50.7%有诱因,治疗前血红蛋白为(121±16)g/L;成人ITP组,102例,男女比例为1∶3.6,74.5%为慢性型,仅15.7%发病前有诱因,治疗前血红蛋白量为(108±30)g/L。两者的性别构成、分型、起病诱因、血红蛋白量比较有统计学意义(P<0.05)。无论儿童还是成人ITP,治疗前骨髓巨核细胞数增多者的疗效明显优于巨核细胞数正常或减少者(均为P<0.05)。随访病例中31例(78%)儿童ITP痊愈,2例(5%)成人ITP痊愈。结论:儿童ITP通常起病急、病情可自行缓解,预后好;成人ITP起病隐袭,多呈慢性经过,病情反复发作。ITP患者治疗前骨髓巨核细胞数增高者比正常或减少者疗效好,检查骨髓巨核细胞数有助于临床判断疗效。     

姜黄素在急性髓性白血病HL—60细胞中，对凋亡调控蛋白Bax、Bak、Mcl—1的影响

为了探讨姜黄素在诱导急性髓性白血病HL－60细胞凋亡时，Bcl-2基因家族成员是否参与了其调控过程。选用Bcl-2基因家族成员Mcl-2、Bax、Bak蛋白为研究对象，SABC法检测其表达状况；用流式细胞术测定DNA直方图中Sub-G1峰值、末端TdT酶标技术检测TUNEL细胞阳性率。发现当25μmol/L姜黄素分别处理HL－60细胞24h、36h、48h后，可使Mcl-1表达下调，Bax和Bak上调，呈时间依赖性。在各处理组，Mcl-1、Bax、Bak表达同时照组相比有显著差异（F值分别为3.443、8.328；P值分别为0.040、0.001）。结果表明：Bcl-2基因家族成员确实参与了姜黄素诱导HL－60细胞凋亡的调控过程。     

家兔动脉粥样硬化与血小板源内皮型氧化氮合酶表达的相关性

Objective To explore the correlation of atheroselerosis progression and the expression of platelet derived endothelial nitric oxide synthase (eNOS) in rabbits. Methods A total of 24 male New Zealand white rabbits were used in this study. Six of the animals were fed with normal food (control group). Eighteen rabbits were fed with cholesterol-rich food (1 g/d) for 12 weeks to establish the atherosclerosis model. Among 18 models, 6 rabbits were executed immediately and their aorta and platelet samples were collected for further analysis (model group), 6 rabbits were orally administered with pravastatin (10 rag/d) for additional 12 weeks (treated group), and the remaining 6 rabbits were left untreated until the end of the study (untreated group). The control, treated and untreated animals were then killed, and the aorta and platelet samples were collected for eNOS expression analysis (RT-PCR). Results The aorta samples in model and untreated group exhibited rough intima and a lot of longitudinal fatty streaks, which indicated that atherosclerosis models were established successfully. While in treated group, the degree of atherosclerosis was decreased. The average percent of thickness of fatty streaks or atheroselerotic plaques relative to the whole thickness of vessel walls was 0. 04±0. 02, 0. 82±0. 16, 0. 33±0. 18,0. 77±0. 14 in control, model, treated and untreated group, respectively. The thickness of fatty streaks or atherosclerotic plaques was significantly increased in the model and untreated groups and decreased in treated group compared with the control group (both P<0. 05). The expressions of platelet derived eNOS/mRNA were 1. 02± 0. 28, 0. 41± 0. 27, 1.00 ± 0. 77, 0. 40±0. 29 in control, model, treated and untreated group, respectively. The expression of eNOS/mRNA was markedly decreased in model group and untreated group compared with the control group, but was increased in treated group compared with untreated and model groups (F=3. 544, P = 0. 024). Conclusions There is a negative correlation between eNOS expression and atherosclerosis development, which suggests that the reversal effect of pravastatin on atheroselerosis progression and plaque formation may relate to the expression of platelet derived eNOS.     

GPⅡb/Ⅲa受体拮抗剂RGDS对血小板聚集和胞浆游离钙的影响

[目的]探讨血小板GPⅡb/Ⅲa受体拮抗剂RGDS对血小板聚集和血小板[Ca2+]i的影响.[方法]比浊法测定PAG;采用Fura-3/AM荧光探针标记血小板胞浆钙离子,使用共聚焦显微镜观察单个血小板荧光强度的变化,计算机图像系统分析血小板[Ca2+]i的变化.[结果]凝血酶(0.03 U/mL)为激动剂时,血小板PAG(M)为(78.2±12.4)%.在所取的62.5～1 000μmol/L RGDS内的5种浓度下,RGDS可呈浓度依赖性地抑制凝血酶诱导的PAG(M).正常人静息血小板[Ca2+]i荧光强度值为376.1±70.5;凝血酶(0.03 U/mL)激动的血小板[Ca2+]i荧光强度值升高为977.9±108.8;GPⅡb/Ⅲa受体拮抗剂RGDS(250 μmol/L)对静息血小板的[Ca2+]i荧光强度值无抑制作用;GPⅡ b/Ⅲa受体拮抗剂RGDS(250μmol/L)作用于血小板后,凝血酶(0.03 U/mL)激动的血小板[Ca2+]i荧光强度值升高受到抑制,抑制率为(9.37±7.5)%.[结论]凝血酶(0.03 U/mL)可以引起血小板的聚集和血小板[Ca2+]i的升高.GP Ⅱb/Ⅲa受体拮抗剂RGDS可以抑制凝血酶(0.03 U/mL)引起的血小板聚集和[Ca2+]i的升高.     

The effect of curcumin on mismatch repair (MMR) proteins hMSH2 and hMLH1 after ultraviolet (UV) irradiation on HL-60 cells

Summary To understand the expression and effect of mismatch repair genes, hMSH2 and hMLH1, and to investigate anti-leukemic cell proliferation mechanism of curcumin, the levels of both genes were detected by multiple comparative RT-PCR. The protein of hMSH2 was determined by flow cytometry (FCM) and the gene mutation of hMSH2 and hMLH1 were detected by PCR-SSCP and microsatellite instability assay. After UV irradiation, the gene expression of hMSH2 and hMLH1 was not increased and showed no response. This phenomenon was not ascribed to gene mutation, because PCP-SSCP and microsatellite instability assay revealed no abnormal gel-shift band in both genes. After irradiation and addition of curcumin, the expression of hMSH2 mRNA increased and the cellular apoptotic rate also increased at the same time. The difference was statistically significant as compared with groups without addition of curcumin and control groups (P<0.05). Our results suggested that when MMR system was inhibited by the same agents, curcumin can remove this suppression and switch to cellular apoptosis. CHEN Yan, male, born in 1952, Professor This project was supported by a grant from the National Natural Sciences Foundation of China (Serial No. 39770934).     

The Effect of Curcumin on Mismatch Repair(MMR) Proteins hMSH2 and hMLH1 after Ultraviolet(UV) Irradiation on HL-60 Cells

To understand the expression and effect of mismatch repair genes, hMSH2 and hMLH1,and to investigate anti-leukemic cell proliferation mechanism of curcumin, the levels of both geneswere detected by multiple comparative RT-PCR. The protein of hMSH2 was determined by flow cy-tometry (FCM) and the gene mutation of hMSH2 and hMLH1 were detected by PCR-SSCP and mi-crosatellite instability assay. After UV irradiation, the gene expression of hMSH2 and hMLH1 wasnot increased and showed no response. This phenomenon was not ascribed to gene mutation, becausePCP-SSCP and microsatellite instability assay revealed no abnormal gel-shift band in both genes. Af-ter irradiation and addition of curcumin, the expression of hMSH2 mRNA increased and the cellularapoptotic rate also increased at the same time. The difference was statistically significant as comparedwith groups without addition of curcumin and control groups (P<0.05). Our results suggested thatwhen MMR system was inhibited by the same agents, curcumin can remo     

HIF—1α基因表达沉默对肝癌细胞P13K／AKT信号转导通路影响的研究

目的：探讨沉默缺氧诱导因子-1α（hypoxiainduciblefactor-1α,HIF-1α）基因表达对肝癌细胞P13K／AKT信号转导通路的影响。方法：选用大鼠CBRH-7919肝癌细胞株作为研究对象,利用氯化钻（CoCl2）建立厌氧模型。构建HIF-1α小分子干扰RNA（smallinterferingRNA,siRNA）特异性载体复合物,小分子RNA干扰技术沉默肝癌细胞HIF-1α的基因表达,分别利用RT-PCR和免疫印迹法检测肝癌细胞HIF-1α、血管内皮生长因子（vascularendothelialgrowthfactor,VEGF）、磷酸化AKT（p-AKT）、AKT和P13K在iTIRNA和（或）蛋白水平的表达变化。结果：肝癌细胞在缺氧条件下表达HIF-1α较常氧对照组显著增多,差异具有统计学意义,PmRNA=0．002,P蛋白=0．003。特异性HIF-1αsiRNA表达载体转染肝癌细胞后,肝癌细胞HIF-1α（PmRNA和P蛋白均〈0．001）、VEGF（PmRNA=0．001,P蛋白〈0．001）和p-AKT（P§自〈0．001）的表达明显减少;AKT和P13K的表达显著增多,与对照组的差异均具有统计学意义,P蛋白值分别为0．032和0．020。结论：特异性siRNA干扰沉默HIF-1α基因表达可抑制肝癌细胞P13K／AKT信号转导通路的激活。