氯沙坦和卡维地洛对缺血再灌注心肌细胞凋亡的影响

目的 :研究氯沙坦和卡维地洛对缺血再灌注心肌细胞凋亡的影响 ,比较氯沙坦和卡维地洛对缺血再灌注心肌损伤的保护作用。方法 :结扎Wistar大鼠左冠状动脉前降支 ,建立大鼠缺血再灌注动物模型 ,采用原位末端标记细胞凋亡法检测心肌细胞凋亡 ,并利用光学显微镜进行细胞计数。结果 :单纯缺血 再灌注组心肌细胞凋亡数较假手术组明显增多 (37.5 3± 9.2 2 /视野∶0 .18± 0 .0 9/视野 ,P <0 .0 5 ) ,氯沙坦和卡维地洛组心肌细胞凋亡数分别为 8.74± 3.5 1/视野和 7.6 3± 4 .0 5 /视野 ,较单纯缺血 再灌注组明显减少 (P <0 .0 5 ) ,氯沙坦和卡维地洛两组间无明显区别 (P >0 .0 5 )。结论 :氯沙坦和卡维地洛对缺血再灌注心肌细胞损伤具有相似的保护作用 ,可明显减少缺血再灌注心肌细胞凋亡     

大鼠心肌缺血/再灌注损伤心肌细胞凋亡与Fas、FasL基因表达的变化

目的：探讨大鼠实验性心肌缺血再灌注时心肌细胞凋亡与Fas及Fas蛋白配体（Fas Ligand,FasL）基因表达的变化及与心肌组织损伤的关系。方法：以穿线结扎或松扎左冠状动脉制备大鼠心肌缺血再灌注模型。64只大鼠随机分成假手术组（假手术24h）、缺血再灌注I组（缺血30min、再灌注24h）、缺血再灌注Ⅱ组（缺备30min、再灌注72h）及缺血再灌注Ⅲ组（缺血3h、再灌注24h）。以缺口末端标记法检测心肌细胞凋亡的变化,S－P免疫组化法分别检测Fas与FasL蛋白水平变化,采用逆转录聚合酶链反应法检测Fas基因mRNA的表达改变,并分析心肌组织病理学损伤程度。结果：心肌缺血再灌注后心肌细胞凋亡指数Fas蛋白阳性染色指数与炎性细胞FasL蛋白阳性染色指数均增加,且均随缺血或再灌注时间延长而进一步增高; Fas基因的mRNA表达也上调,但以再灌注24h时达高峰;心肌缺血再灌注后心肌组织呈大小不一的灶性坏死,坏死周围有爆炸性一细胞浸润。结论：心肌缺血再灌注时心肌细胞凋亡、Fas基因的蛋白与mRNA表达水平及炎性细胞的FasL蛋白表达量均增加,心肌细胞凋亡与Fas/FasL系统参与了心肌缺血再灌注损伤过程。     

大鼠急性心肌缺血再灌注损伤诱导细胞凋亡的实验研究

目的:观察心肌缺血再灌注(IR)损伤与心肌细胞凋亡关系及凋亡调控基因Bcl-2、Bax表达情况。方法:选用健康雄性SD大鼠29只,随机分为:假手术组(n=8),缺血组(n=12),缺血再灌注组(n=9),分别取上述大鼠心肌组织。(1)观察心肌组织及线粒体的形态结构,并进行体视学分析。(2)采用缺口末端标记法(TUNEL)原位检测凋亡细胞。(3)用免疫组化SP法检测心肌细胞Bcl-2、Bax表达。结果:(1)假手术组心肌纤维排列整齐,细胞间质血管未见明显异常,细胞核膜完整,缺血组及缺血再灌注组可见心肌嗜酸性变、空泡变性、心肌纤维紊乱及收缩带形成,心肌纤维间出血及灶性心肌坏死。心肌细胞线粒体体视学分析,与假手术组比较,心肌缺血组形状因子显著降低(P<0.05),面密度显著增加(P<0.05),缺血再灌注组形状因子显著降低(P<0.01),面密度及周密度均显著增加(均P<0.05)。(2)与假手术组比较,缺血组细胞凋亡率明显升高(P<0.001),缺血再灌注组细胞凋亡率明显升高(P<0.001)。缺血再灌注组与缺血组比较细胞凋亡率明显升高(P<0.05)。(3)与假手术组比较,缺血再灌注组凋亡调控基因Bcl-2、Bax表达明显升高(P<0.01,P<0.001)。缺血再灌注组与缺血组比较凋亡调控基因Bcl-2、Bax表达明显升高(均P<0.01)。结论:心肌缺血及再灌注在导致细胞形态、线粒体超微结构改变的同时,诱导心肌细胞的凋亡,Bcl-2、Bax蛋白表达在心肌细胞凋亡的发生中起重要作用,细胞凋亡加重了缺血再灌注损伤。     

卡维地洛对大鼠缺血再灌注心肌细胞凋亡及基因表达的影响

目的 :研究卡维地洛对缺血再灌注心肌细胞凋亡及bcl 2、bax基因表达的影响 ,探讨卡维地洛抑制缺血再灌注心肌细胞凋亡的可能分子机制。方法 :结扎Wistar大鼠左冠状动脉前降支 (LAD) ,建立大鼠缺血再灌注动物模型。 30只大鼠分 3组 (每组 10只 ) :卡维地洛组 (卡维地洛治疗 )、缺血再灌注组和假手术组。用Tunnel法检测心肌细胞凋亡 ,并用光学显微镜进行细胞计数 ,免疫组化和原位杂交法检测bcl 2、bax基因表达 ,并利用图像分析系统检测二者的平均光密度值 (A值 ) ,进行定量分析。结果 :心肌细胞凋亡数在缺血再灌注组、假手术组和卡维地洛组分别为 36 .8± 9.0、0 .2± 0 .1和 9.5± 3.0 ,各组间差异有统计学意义 (P <0 .0 1)。缺血再灌注组、假手术组和卡维地洛组的bcl 2mRNAA值分别为 0 .0 6± 0 .0 1、0 .0 8± 0 .0 1和 0 .0 9± 0 .0 1,bcl 2蛋白平均A值分别为0 .0 8± 0 .0 2、0 .14± 0 .0 1和 0 .15± 0 .0 3,卡维地洛组与缺血再灌注组相比差异无统计学意义 (P >0 .0 5 ) ;缺血再灌注组bax蛋白的平均A值为 0 .13± 0 .0 2 ,假手术组为 0 .0 7± 0 .0 1,卡维地洛组为 0 .0 6± 0 .0 1,卡维地洛组与缺血再灌注组相比差异有统计学意义 (P <0 .0 5 ) ,bcl 2 /bax蛋白比值在缺血再灌注组、假手术组和卡     

硝酸甘油对心肌细胞凋亡和Fas基因蛋白及mRNA表达的影响

目的　探讨硝酸甘油对持续缺血和缺血再灌注损伤时心肌细胞凋亡和 Fas基因蛋白及 m RNA表达的影响。方法　将大白鼠随机分为 5组 ,以活结结扎左冠状动脉的前降支 ,分别造成阻断冠脉血流和再灌注。以缺口末端标记法 (TUNEL)标记凋亡细胞 ;以 S- P免疫组化及逆转录聚合酶链反应 (RT- PCR)法分别检测 Fas基因蛋白与 m RNA的表达变化。结果　凋亡细胞原位标记与半定量分析表明 :硝酸甘油处理组与持续缺血组、缺血再灌注组比较心肌细胞凋亡程度明显降低 ,Fas基因蛋白及 m RNA表达明显减少。结论　硝酸甘油预处理能能明显减轻心肌细胞凋亡程度 ;其机制可能与下调 Fas基因蛋白及 m RNA表达有关。     

丹参注射液对大鼠心肌缺血再灌注时心肌细胞凋亡的影响

目的 :研究丹参注射液对大鼠心肌缺血再灌注时心肌细胞凋亡的影响。方法 :用在体左冠状动脉前降支穿线结扎法制备心肌缺血再灌注模型。 6 0只SD大鼠分为 3组 :假手术组 (10只 ) ,缺血再灌注组 (2 5只 ) ,丹参注射液干预组 (2 5只 )。缺血 30min ,再灌注 12 0min。采用缺口末端标记法 (TUNEL)以及S P免疫组化法 ,检测心肌细胞凋亡指数 (AI)、Bcl 2、Bax基因蛋白表达。结果 :假手术组AI 2 .2± 0 .6 ,Bcl 2OD值 0 .112± 0 .0 18,BaxOD值 0 .10 2± 0 .0 31;缺血再灌注组AI 2 8.9± 3.6 ,Bcl 2OD值 0 .0 92± 0 .0 14 ,BaxOD值 0 .136± 0 .0 2 2 ;丹参注射液干预组AI 2 6 .2± 3.4 ,Bcl 2OD值 0 .10 4± 0 .0 2 1,BaxOD值 0 .12 2± 0 .0 12。与假手术组比较 ,缺血再灌注组AI增加 ,Bax阳性表达增强 ,Bcl 2阳性表达减弱 (P <0 .0 1) ;与缺血再灌注组比较丹参注射液干预组 ,AI、Bax阳性表达下降 (P <0 .0 1) ,Bcl 2阳性表达上调 (P <0 .0 5 )。结论 :丹参注射液通过抑制心肌细胞凋亡 ,下调Bax基因蛋白表达 ,上调Bcl 2基因蛋白表达 ,起到心肌缺血再灌注损伤的保护作用。     

N-乙酰半胱氨酸对家兔缺血再灌注损伤心肌细胞凋亡的影响

目的观察N-乙酰半胱氨酸(NAC)对家兔缺血再灌注损伤心肌细胞凋亡的影响,并探讨其机制。方法 30只家兔随机等分为假手术组、心肌缺血再灌注(I/R)组和NAC组。NAC组和I/R组制作缺血再灌注模型,前者在血流灌注后阻断主动脉和肺动脉血管10 s,同时于心尖部注入NAC溶液0.5 mL;I/R组在再灌注开始即刻给予生理盐水0.5 mL。假手术组家兔开胸后只穿线,不结扎冠状动脉左前降支。再灌注3 h后,用黄嘌呤氧化酶法测定血清超氧化物歧化酶(SOD)活性,硫代巴比妥酸法测定血清丙二醛(MAD)水平。三组各取缺血区心肌组织,用原位末端标记法测定心肌细胞凋亡,并计算凋亡指数(AI)。用免疫组化SABC法检测缺血心肌组织中的Fas蛋白,计算Fas蛋白阳性表达指数(PI)。结果 I/R组的MDA水平高于、SOD活性低于NAC组和假手术组(P均<0.05);I/R组缺血区心肌细胞AI、心肌细胞中Fas蛋白PI明显高于NAC组和假手术组(P均<0.05)。结论 NAC可减少家兔缺血再灌注损伤心肌细胞的凋亡,可能是通过其抗氧化作用及下调Fas蛋白表达来实现的。     

人参皂苷Re抑制心肌缺血再灌注损伤中性粒细胞浸润和髓过氧化物酶活性的研究

目的 :观察人参皂苷Re对心肌缺血再灌注中性粒细胞 (PMNs)浸润及髓过氧化物酶 (MPO)活性的影响 ,探讨人参皂苷Re对心肌缺血再灌注损伤的防治作用。方法 :结扎SD大鼠左冠状动脉前降支 (LAD) ,建立大鼠缺血再灌注动物模型 ;苏木精 伊红染色光镜检测心肌组织中PMNs浸润的数量 ;MPO法测定心肌MPO活性。结果 :①心肌缺血再灌注后 ,光镜下假手术组未发现PMNs ,缺血再灌注组和人参皂苷Re治疗组 (简称Re治疗组 )PMNs浸润明显增加。缺血再灌注组PMNs数为 (8.16 6 7± 1.14 72 )个 /视野 ;Re治疗组为 (4 .5 0 0 0±1.0 4 88)个 /视野 ;两者与假手术组比较差异有统计学意义 (P <0 .0 1) ;Re治疗组与缺血再灌注组相比 ,PMNs浸润明显减少 ,差异有统计学意义 (P <0 .0 1)。②缺血再灌注组、Re治疗组MPO活性均较假手术组明显增加(P <0 .0 1) ,Re治疗组与缺血再灌注组相比MPO活性明显降低 (P <0 .0 1)。结论 :心肌缺血再灌注诱导PMNs的浸润和MPO活性的增加 ,人参皂苷Re可显著抑制心肌缺血再灌注PMNs浸润和MPO活性     

缺血预处理对心肌缺血再灌注诱发凋亡的保护机制

目的：通过观察心肌细胞凋亡指数与心肌中Fas、C-fOS表达阳性率的变化,探讨缺血预处理对心肌缺血再灌注诱发细胞凋亡的保护作用及与心肌中Fas、c-fos基因表达的关系. 方法：将入选大鼠随机分为假手术对照组（A组）、缺血组（B组）、缺血再灌注组（C组）、缺血预处理组（D组）,每组15只,测定各组大鼠心肌中细胞凋亡和Fas、c-fos的表达,并分析三者之间的相关性. 结果：与A组比较,B、C及D组大鼠的心肌凋亡指数、心肌中Fas与c-fos的阳性率均升高（P＜0.01）;与C组比较,B组及D组大鼠的心肌凋亡指数、心肌中Fas与C-fOS的阳性率均降低（P＜0.01）.Pearson相关分析显示心肌凋亡指数、Fas、c-fos三者之间存在显著正相关. 结论：缺血预处理可能通过下调凋亡相关基因Fas、c-fos的表达,减少细胞凋亡而发挥心肌保护作用.     

缺血-再灌注诱导家兔房室结细胞凋亡的研究

为探讨缺血 再灌注对在体家兔房室结细胞凋亡的作用 ,以及凋亡相关调控基因蛋白 (Fas、bcl 2、bax)的表达 ,以家兔为实验对象 ,麻醉后开胸暴露心脏 ,结扎右冠状动脉 ,建立房室结缺血 再灌注模型。于预定时间处死动物 ,快速取出房室交界区组织 (3mm× 2mm× 1mm) ,10 %福尔马林磷酸盐缓冲液固定。采用TUNEL法检测房室结细胞凋亡 ;免疫组化法检测Fas、bcl 2、bax基因蛋白表达。结果 :①对照组及单纯缺血 10 ,30min组均未检测到细胞凋亡 ,缺血 6 0min组可见少量散在的凋亡细胞 ,缺血 12 0min房室结可见大量的细胞凋亡 ;②缺血 再灌注组 :不同缺血时间组均出现细胞凋亡 ,凋亡细胞数随缺血时间的延长而增加 (P <0 .0 5 ) ;③随缺血时间的延长 ,Fas、bax的表达明显增加 (P <0 .0 1) ,而bcl 2的表达则无明显增加 (P >0 .0 5 ) ;④缺血 再灌注组较相应时间单纯缺血组细胞凋亡数、Fas、bax表达均明显增加 (P <0 .0 1)。结论 :单纯缺血及缺血 再灌注均可诱导房室结细胞发生凋亡 ,这可能与该条件下Fas、bax基因蛋白的过量表达有关。     

妊娠时间与肺结核复发的相关性研究及诊治对策

目的 分析肺结核史患者妊娠时间和肺结核复发间相关性.方法 选取我院收治的有肺结核史的妊娠妇女576例作为研究对象,对其妊娠前肺结核治疗、治愈后妊娠时间、妊娠后复发肺结核等进行分析,总结有肺结核史育龄女性的妊娠时间和肺结核复发之间的关系.结果 肺结核治愈后不同时间段妊娠者的结核复发率比较,差异具有显著性（P＜0.05）,停药后间隔时间越久妊娠,肺结核复发的几率越小.结论 加强孕期痰菌检查,及早发现复发肺结核,提高母婴安全.     

我国骨关节结核诊治的回顾与展望

骨关节结核是危害人们健康的严重感染性疾病,近95％由他处结核病继发而来.罹患骨关节结核疾病后几乎均将致残,严重影响人们的健康、工作和生活.建国以来在党和国家的关心和支持下,骨关节结核的诊治水平取得了长足进步.时至今日,由于多种原因,学科发展和被重视程度受到一定的制约,同整个医疗行业的发展不相适应.回顾过去,展望未来,我们需要重新审视骨关节结核的诊治方法,努力推进骨关节结核诊疗技术的科学发展.     

Effect of phosphorylation of MAPK and Stat3 and expression of c-fos and c-jun proteins on hepatocarcinogenesis and their clinical significance

AIM To study the effect of phosphorylation ofMAPK and Stat3 and the expression of c-fos andc-jun proteins on hepatocellular carcinogenesisand their clinical significance.METHODS SP immunohistochemistry was usedto detect the expression of p42/44~(MAPK), p-Stat3,c-fos and c-jun proteins in 55 hepatocellularcarcinomas (HCC) and their surrounding livertissues.RESULTS The positive rates and expressionlevels of p42/44~(MAPK), p-Stat3, c-fos and c-junproteins in HCCs were significantly higher thanthose in pericarcinomatous liver tissues (PCLT).A positive correlation was observed between theexpression of p42/44~(MAPK) and c-fos proteins, andbetween p-Stat3 and c-jun, but there was nosignificant correlation between P42/44~(MAPK) and p-Stat3 in HCCs and their surrounding livertissues.CONCLUSION The abnormalities of Ras/Raf/MAPK and JAKs/ Stat3 cascade reaction maycontribute to malignant transformation ofhepatocytes. Hepatocytes which are positive forp42/ 44~(MAPK), c-fos or c-jun proteins may bepotential malignant pre-cancerous cells.Activation of MAPK and Stat3 proteins may be anearly event in hepatocellular carcinogenesis.     

Overweight and Obesity and Progression of ADPKD

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Replication and Inhibitors of Enteroviruses and Parechoviruses

The Enterovirus (EV) and Parechovirus genera of the picornavirus family include many important human pathogens, including poliovirus, rhinovirus, EV-A71, EV-D68, and human parechoviruses (HPeV). They cause a wide variety of diseases, ranging from a simple common cold to life-threatening diseases such as encephalitis and myocarditis. At the moment, no antiviral therapy is available against these viruses and it is not feasible to develop vaccines against all EVs and HPeVs due to the great number of serotypes. Therefore, a lot of effort is being invested in the development of antiviral drugs. Both viral proteins and host proteins essential for virus replication can be used as targets for virus inhibitors. As such, a good understanding of the complex process of virus replication is pivotal in the design of antiviral strategies goes hand in hand with a good understanding of the complex process of virus replication. In this review, we will give an overview of the current state of knowledge of EV and HPeV replication and how this can be inhibited by small-molecule inhibitors.     

Effect of phosphorylation of MAPK and Stat3 and expression of c-fas and c-jun proteins on hepatocarcinogenesis and their clinical significance

AIM To study the effect of phosphorylation ofMAPK and Stat3 and the expression of c-fos andc-jun proteins on hepatocellular carcinogenesisand their clinical significance.METHODS SP immunohistochemistry was usedto detect the expression of p42/44MAPK, p-Stat3,c-fos and c-jun proteins in 55 hepatocellularcarcinomas (HCC) and their surrounding livertissues.RESULTS The positive rates and expressionlevels of p42/44MAPK, p-Stat3, c-fos and c-junproteins in HCCs were significantly higher thanthose in pericarcinomatous liver tissues (PCLT).A positive correlation was observed between theexpression of p42/44MAPK and c-fos proteins, andbetween p-Stat3 and c-jun, but there was nosignificant correlation between p42/44MAPK and p-Stat3 in HCCs and their surrounding livertissues.CONCLUSION The abnormalities of Ras/Rat/MAPK and JAKs/ Stat3 cascade reaction maycontribute to malignant transformation ofhepatocytes. Hepatocytes which are positive forp42/ 44MAPK, c-fos or c-jun proteins may bepotential malignant pre-cancerous cells.Activation of MAPK and Stat3 proteins may be anearly event in hepatocellular carcinogenesis.     

心电图、心电向量图与冠状动脉造影结果对比分析

目的:通过分析心电图(Electrocardiogram,ECG)和心电向量图(Vectorcardiogram,VCG)的改变与冠脉造影（CAG）结果进行对比,探讨ECG、VCG在冠状动脉病变中的诊断价值。方法: 选择2008年1月~2009年12月临床拟诊断为冠心病患者108例,行常规ECG、VCG检查,并于1周内进行CAG,对检查结果依据各自的诊断标准进行判定,以CAG为标准诊断法,利用四格表法,计算相关评价真实性的指标并进行比较。结果: ①VCG检测的灵敏度、特异度、准确度显著高于ECG(P<0.05,P<0．01)。②ECG、VCG阳性率与冠脉病变支数组间比较:在单支病变、双支病变中,VCG阳性率明显高于ECG(P<0.05),左主干或三支病变无统计学意义;组内比较:ECG组左主干或三支病变组较单支病变、双支病变阳性率高(P<0.05,P<0.01);VCG组左主干或三支病变组较单支病变阳性率高(P<0.05);与双支病变阳性率比较无统计学意义;③ECG、VCG阳性率与冠脉病变程度组间比较:冠脉病变狭窄50%～69%的VCG阳性率明显高于ECG (P<0.05),其他两组阳性率比较无统计学意义;组内比较:ECG组冠脉病变狭窄≥90%较50%～69%、70%～89%的阳性率高(P<0.05,P<0.01); VCG组狭窄≥90%较50%～69%阳性率高（P<0.01),其他无统计学意义。结论: VCG对冠心病检测价值显著高于ECG。     

Detection and characterization of the product of hydroethidine and intracellular superoxide by HPLC and limitations of fluorescence

Here we report the structural characterization of the product formed from the reaction between hydroethidine (HE) and superoxide (O(2)(.-)). By using mass spectral and NMR techniques, the chemical structure of this product was determined as 2-hydroxyethidium (2-OH-E(+)). By using an authentic standard, we developed an HPLC approach to detect and quantitate the reaction product of HE and O(2)(.-) formed in bovine aortic endothelial cells after treatment with menadione or antimycin A to induce intracellular reactive oxygen species. Concomitantly, we used a spin trap, 5-tert-butoxycarbonyl-5-methyl-1-pyrroline N-oxide (BMPO), to detect and identify the structure of reactive oxygen species formed. BMPO trapped the O(2)(.-) that formed extracellularly and was detected as the BMPO-OH adduct during use of the EPR technique. BMPO, being cell-permeable, inhibited the intracellular formation of 2-OH-E(+). However, the intracellular BMPO spin adduct was not detected. The definitive characterization of the reaction product of O(2)(.-) with HE described here forms the basis of an unambiguous assay for intracellular detection and quantitation of O(2)(.-). Analysis of the fluorescence characteristics of ethidium (E(+)) and 2-OH-E(+) strongly suggests that the currently available fluorescence methodology is not suitable for quantitating intracellular O(2)(.-). We conclude that the HPLC/fluorescence assay using HE as a probe is more suitable [corrected] for detecting intracellular O(2)(.-).     

Effects of sex and time of day on metabolism and excretion of corticosterone in urine and feces of mice

Non-invasive techniques to monitor stress hormones in small animals like mice offer several advantages and are highly demanded in laboratory as well as in field research. Since knowledge about the species-specific metabolism and excretion of glucocorticoids is essential to develop such a technique, we conducted radiometabolism studies in mice (Mus musculus f. domesticus, strain C57BL/6J). Each mouse was injected intraperitoneally with 740 kBq of 3H-labelled corticosterone and all voided urine and fecal samples were collected for five days. In a first experiment 16 animals (eight of each sex) received the injection at 9 a.m., while eight mice (four of each sex) were injected at 9 p.m. in a second experiment. In both experiments radioactive metabolites were recovered predominantly in the feces, although males excreted significantly higher proportions via the feces (about 73%) than females (about 53%). Peak radioactivity in the urine was detected within about 2h after injection, while in the feces peak concentrations were observed later (depending on the time of injection: about 10h postinjection in experiment 1 and about 4h postinjection in experiment 2, thus proving an effect of the time of day). The number and relative abundance of fecal [3H]corticosterone metabolites was determined by high performance liquid chromatography (HPLC). The HPLC separations revealed that corticosterone was extensively metabolized mainly to more polar substances. Regarding the types of metabolites formed, significant differences were found between males and females, but not between the experiments. Additionally, the immunoreactivity of these metabolites was assessed by screening the HPLC fractions with four enzyme immunoassays (EIA). However, only a newly established EIA for 5alpha-pregnane-3beta,11beta,21-triol-20-one (measuring corticosterone metabolites with a 5alpha-3beta,11beta-diol structure) detected several peaks of radioactive metabolites with high intensity in both sexes, while the other EIAs showed only minor immunoreactivity. Thus, our study for the first time provides substantial information about metabolism and excretion of corticosterone in urine and feces of mice and is the first demonstrating a significant impact of the animals' sex and the time of day. Based on these data it should be possible to monitor adrenocortical activity non-invasively in this species by measuring fecal corticosterone metabolites with the newly developed EIA. Since mice are extensively used in research world-wide, this could open new perspectives in various fields from ecology to behavioral endocrinology.     

大鼠骨髓间充质干细胞的分离培养和外源基因的导入

目的探讨绿色荧光蛋白基因转染骨髓间质干细胞的可行性。方法采用F icoll-PaqueTMP lus淋巴细胞分离液,根据细胞密度梯度原理,分离大鼠骨髓间充质干细胞(rM SC s)并进行体外原代培养和传代扩增,倒置相差显微镜观察细胞生长情况,免疫细胞化学法对其初步鉴定。流式细胞仪分析转染效率。结果原代和传代培养的细胞呈现梭形外观,具有较强的生长增殖能力;细胞均一表达CD44、CD54、CD106、CD29抗原。电穿孔法转染rM SC s转染率为32.8%±3%。结论采用比重为1.077 g/L的F icoll-PaqueTMP lus能分离获得大鼠骨髓间充质干细胞,经原代培养和传代培养能够迅速扩增。电穿孔法具有较高的介导外源基因表达于rM SC s的效率。