β受体阻滞剂卡维地洛、美托洛尔对心力衰竭大鼠体液因子及ANP、TNF-α、IL-6、NE的影响

目的分析β受体阻滞剂卡维他洛、美托洛尔对心力衰竭(心衰)大鼠体液因子及血清心钠素(ANP)、肿瘤坏死因子(TNF)-α、白细胞介素(IL)-6、去甲肾上腺素(NE)的影响。方法从军事医学院动物实验中心购买80只大鼠,建立心衰模型,用随机数字法分为假手术组、安慰剂组、卡维地洛组、美托洛尔组各20只,观察6 w,给予血流动力学、心脏超声等检查分析NE、IL-6、TNF-α、血管紧张素(Ang)Ⅱ、ANP等指标。结果假手术组左室短轴缩短率(FS)、左室最大下降速率(-dp/dt)、左室最大上升速率(+dp/dt)、左室收缩末压(LVESP)、平均血压(MBP)、心率(HR)显著高于其他3组,左室舒张末压(LVEDP)显著低于其他3组(P0. 05)。假手术组NE、IL-6、TNF-α、AngⅡ、ANP水平显著高于其他3组(P0. 05);安慰剂组IL-6、TNF-α、NE、ANP水平显著高于卡维地洛组和美托洛尔组(P0. 05);卡维地洛组IL-6、AngⅡ水平显著高于美托洛尔组(P0. 05)。假手术组左室质量指数(LVWI)、室间隔厚度(VSD)、左室后壁厚度(LVPWD)、左室收缩末期直径(LVESD)、左室舒张末期直径(LVEDD)显著低于其他3组(P0. 05);卡维地洛组和美托洛尔组LVWI、VSD显著低于安慰剂组(P0. 05),LVESD、LVWI、VSD显著高于假手术组(P0. 05)。结论β受体阻滞剂可更为全面降低其细胞因子、神经内分泌指标,可能因其具备改善心功能作用,并对左室重塑产生抑制。而卡维地洛和美托洛尔药物对比,前者在降低细胞因子、神经内分泌、心肌重塑等方面作用更为显著。     

美托洛尔和卡维地洛对慢性心力衰竭的影响

目的比较选择性β1受体阻滞剂美托洛尔和非选择性β受体阻滞剂卡维地洛治疗对慢性心力衰竭(CHF)代谢底物、细胞因子及心脏功能的影响。方法选择CHF患者86例(CHF组)及健康体检者25例(正常对照组)。CHF组患者又随机分为美托洛尔组(43例)和卡维地洛组(43例)。记录两组CHF患者治疗前后心功能分级及不良事件次数及TNF-α、白细胞介素-1β(IL-1β)和白细胞介素-6(IL-6)含量。所有入选者均测定血清游离脂肪酸(FFA)含量。结果美托洛尔组和卡维地洛组患者经过治疗后心功能明显改善(P<0.01),其中卡维地洛组更加明显(P<0.01)。美托洛尔组不良事件40次,卡维地洛组不良事件24次(P<0.01)。血浆TNF-α、IL-1β和IL-6较治疗前显著降低(P<0.05)。卡维地洛组较美托洛尔组TNFα和IL-1β降低更明显(P<0.01,P<0.05)。CHF组患者血清FFA含量同正常对照组比较明显升高(P<0.01)。治疗后卡维地洛组血清FFA较美托洛尔组降低更明显(P<0.01)。结论β受体阻滞剂可以改善CHF患者心功能,降低血浆细胞因子及FFA水平,非选择性的β受体阻滞剂卡维地洛优于选择性的β1受体阻滞剂美托洛尔。     

β受体阻滞剂卡维地洛改善慢性心衰患者的舒张功能及二尖瓣返流

几项研究表明β受体阻滞剂可改善慢性心衰者左室工作,提高心脏收缩力,降低病残率和死亡率。卡维地洛(Carvedilol)是一种β受体阻滞剂,兼有α_1血管舒张及促进新陈代     

卡维地洛与美托洛尔治疗心力衰竭的疗效对比观察

目的:比较卡维地洛与美托洛尔治疗充血性心力衰竭(CHF)的疗效及对神经激素、细胞因子的影响。方法:选择CHF患者120例,随机分3组。A组为对照组:予以血管扩张剂、利尿剂、地高辛等常规心力衰竭治疗。B、C组在上述治疗基础上分别予以美托洛尔50mg,bid、卡维地洛25mg,bid口服,维持该剂量至6个月。用药前后分别观察左室射血分数(LVEF)、每搏输血量(SV)、短轴缩短率(FS)、左室舒张末期内径(LVEDD)、左室收缩末期内径(LVESD)、相对室壁厚度(RWT)及醛固酮(ALD)、肿瘤坏死因子-α(TNF-α)、内皮素(ET)、心钠素(ANP)等指标变化情况。结果:6个月后B组及C组LVEDD、LVESD缩小,RWT增厚,LVEF、FS、SV明显提高,而ALD、ET、TNFα、ANP水平明显降低,与治疗前及A组比较均差异有统计学意义。B、C组再入院率及病死率均明显低于A组,同时C组LVEF改善优于B组。结论:美托洛尔、卡维地洛均可明显降低CHF患者神经激素、细胞因子的水平,逆转心室重塑,改善心脏功能。卡维地洛疗效及耐受性略优于美托洛尔。     

卡维地洛对老年慢性心力衰竭合并室性心律失常患者血清肾上腺素受体自身抗体的影响

目的 研究卡维地洛对老年慢性心力衰竭(心衰)合并室性心律失常(室律失常)患者血清抗β1、β2和α1肾上腺素受体自身抗体的影响.方法 将68例老年冠心病心衰合并有室律失常的患者随机分为两组,在常规强心利尿治疗的同时,一组给予美托洛尔、另一组给予卡维地洛治疗,检测两组治疗前和治疗后6个月心脏超声、B型利钠肽(BNP)、动态心电图和抗β1、β2和α1受体自身抗体阳性率的改变,同时检测治疗前后血压、心率、肝和肾功能的变化.结果 与治疗前相比,两组均使心衰患者的基础心率和BNP下降,左室射血分数(LVEF值)升高,心脏功能得到改善;与美托洛尔组相比,卡维地洛治疗后收缩压和BNP下降更明显,差异有统计学意义(P<0.01).美托洛尔治疗后,可使患者血清中抗β1受体自身抗体的阳性率下降(P<0.05),而对抗β2和α1 受体自身抗体的阳性率没有影响(P>0.05).卡维地洛治疗后,血清中抗β1、β2和α1受体自身抗体的阳性率均明显下降(P<0.01),室律失常的发生率也比美托洛尔治疗组明显下降(P<0.01).结论 对老年冠心病慢性心衰合并室律失常患者,应用卡维地洛比美托洛尔能更有效地降低室律失常的发生.     

卡维地洛对心肌梗死后大鼠心肌细胞因子表达及心室重塑的影响

目的探讨细胞因子在心肌梗死(MI)后大鼠心室重塑中的作用,以及卡维地洛对心肌细胞因子表达和心室重塑的影响。方法结扎大鼠左冠状动脉前降支建立MI模型,随机分为对照组和卡维地洛组,另设假手术组。卡维地洛组给予卡维地洛灌胃,对照组和假手术组仅以等体积生理盐水灌胃,观察4周末卡维地洛对心肌细胞肿瘤坏死因子-α(TNF-α)、白介素-1β(IL-1β)、白介素-6(IL-6)、转化生长因子β1(TGF-β1)和白介素-10(IL-10)mRNA的表达及心室重塑的影响。结果MI后4周末大鼠心肌细胞因子表达(梗死区:TNF-α1.07±0.23、IL-1β1.07±0.36、IL-61.25±0.47、TGF-β11.25±0.16I、L-100.84±0.06;非梗死区:TNF-α1.05±0.09I、L-1β1.04±0.14、IL-61.04±0.13、TGF-β11.05±0.07I、L-100.77±0.09)较假手术组明显增加(P<0.01)。与对照组比较,卡维地洛组血液动力学指标及心室重塑改善,心肌致炎性细胞因子mRNA表达(梗死区:TNF-α0.58±0.06I、L-1β0.71±0.08I、L-60.63±0.08;非梗死区TNF-α0.67±0.19、IL-1β0.58±0.11、IL-60.59±0.15)和致纤维化性细胞因子TGF-β1mRNA表达(梗死区0.72±0.14,非梗死区0.73±0.16)降低,而抗炎性细胞因子IL-10mRNA表达差异无统计学意义。结论细胞因子可能参与了MI后心室重塑。卡维地洛改善心室重塑的作用机制至少部分与对细胞因子基因表达作用有关。     

慢性心力衰竭患者外周血中血浆肾素活性和利钠肽水平的变化及卡维地洛对其影响

目的　观察慢性心力衰竭 (心衰 )患者外周血中血浆肾素活性 (PRA)、心钠肽 (ANP)及脑钠肽 (N BNP)水平的变化及卡维地洛对其影响。方法　 6 0例慢性心衰患者随机分为常规治疗组 (血管紧张素转换酶抑制 +利尿剂 +地高辛 )和卡维地洛组 (常治疗药物 +卡维地洛 ) ,随访 12w ,采用放射免疫法测定二组治疗前后和 30例健康体检者 (正常对照组 )外周血中PRA、ANP、及N BNP水平。同时使用核素心室显像测定心衰患者左心室射血分数 (LVEF)。结果　心衰患者外周血中PRA、ANP及N BNP水平较正常对照组显著升高 ,其中ANP及N BNP水平在卡维地洛治疗前与LVEF负相关 ,在卡维地洛治疗后与LVEF密切相关 ,但PRA水平与LVEF无关。治疗后卡维地洛组外周血中PRA、ANP及N BNP水平较常规治疗组下降更明显。结论　外周血中ANP及N BNP水平在慢性心衰的病理生理机制中起着重要作用 ,甚至在 β受体阻滞剂治疗后仍可用于指导心衰患者的治疗。β受体阻滞剂能抑制心衰患者神经内分泌的过度激活。     

β受体阻断剂对慢性心力衰竭患者血浆vWF及TNF-α的影响

目的:观察充血性心力衰竭(CHF)患者口服β受体阻断剂美托洛尔和卡维地洛后血浆血管性假血友病因子(vWF)及肿瘤坏死因子-α(TNF-α)的改变,并探讨其改变的临床意义。方法:63例CHF患者随机给予美托洛尔或卡维地洛达目标剂量后12周,观察血浆vWF及TNF-α浓度的变化。30例体检正常健康者作为对照组。血浆vWF采用酶标法测定,TNF-α采用生物活性法测定。结果:2组CHF患者基础血浆vWF和TNF-α浓度高于健康对照组(P<0.01);服药4周后,美托洛尔组和卡维地洛组血浆vWF和TNF-α浓度下降(P<0.05),12周后进一步下降(P<0.01)。结论:β受体阻断剂能下调CHF患者血浆vWF及TNF-α浓度。     

卡维地洛对慢性心力衰竭患者心肌重构和炎性细胞因子的影响

目的探讨卡维地洛对慢性心力衰竭(心衰)患者心肌重构和血浆肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)和白细胞介素-6(IL-6)的影响。方法72例心衰患者随机分为治疗组和对照组各36例。对照组给予洋地黄、利尿剂及血管紧张素转换酶抑制剂。治疗组在此基础上加用卡维地洛。随访半年,治疗前后采用超声心动图、胸部X线和化学发光法测定心功能、心胸比率和血浆TNF-α、IL-1β和IL-6的变化。结果两组治疗后心功能改善,总有效率为77.8%和50%,P<0.01;心胸比率分别为(0.55±0.07和0.58±0.06,P<0.05)。治疗组治疗后左室舒张期末内径和左室收缩期末内径分别为(58.7±6.9)mm和(44.7±7.2)mm,显著低于对照组(61.3±2.7)mm和(50.9±9.2)mm;左室射血分数为(46.3±9.7)%,显著高于对照组(39.9±6.8)%,P<0.01;血浆TNF-α、IL-1β和IL-6分别为(46.08±13.27)pg/ml、(31.32±4.06)pg/ml和(30.26±10.59)pg/ml,显著低于对照组(59.23±18.65)pg/ml、(39.58±11.38)pg/ml和(54.96±20.21)pg/ml,P<0.01)。副作用发生率,两组相比较差异无统计学意义。结论慢性心力衰竭在常规治疗基础上加用卡维地洛安全有效,可以改善心功能,逆转心肌重构,降低血浆主要细胞因子水平。     

不同肾上腺素能受体阻滞剂对改善压力负荷大鼠左室重塑的作用

目的　比较卡维地洛、美托洛尔和特拉唑嗪对压力超负荷性左室心肌胶原结合蛋白(collagen bindingprotein ,Colligin)和肌球蛋白同工酶 (α/ β MHC)表达的影响 ,探讨卡维地洛改善左室重塑的新机制。方法　将腹主动脉缩窄术后 4周的雄性Wistar大鼠随机分为腹主动脉缩窄术、卡维地洛、美托洛尔、特拉唑嗪 4组 (n =8)。治疗 12周后 ,观察各组大鼠各项指标的变化。结果　术后 16周大鼠左室明显肥厚 ,α/ β MHCmRNA的比值下降 ,ColliginmRNA表达及Ⅰ /Ⅲ型胶原和Colligin蛋白含量增加 (P <0 0 5 ) ;药物治疗 12周后 ,卡维地洛能够明显改善左室肥厚 ,美托洛尔次之 ,而特拉唑嗪作用不明显 ;卡维地洛组大鼠左室心肌Ⅰ /Ⅲ型胶原和Colligin蛋白的表达下降 ,α/ β MHCmRNA表达比值增加 ,ColliginmRNA表达下调 (P <0 0 5 ) ,但美托洛尔组和特拉唑嗪组无此变化 (P >0 0 5 )。结论　卡维地洛与美托洛尔均能明显改善左室肥厚 ,而非选择性 β肾上腺素能受体阻滞剂卡维地洛更能下调心室细胞外基质蛋白和逆转肌球蛋白同工酶转换 ,其作用机制有待进一步确定。     

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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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.     

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.     

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.     

荣宝和氯硝柳胺灭螺效果比较及成本分析

目的 评价新型灭螺药物荣宝杀灭钉螺的效果,探讨其推广应用价值.方法 按目前推荐的荣宝灭螺剂量,喷洒法为30 g/m2,浸杀法为50 g/m3;氯硝柳胺喷洒法和浸杀法分别采用2 g/m2和2 g/m3杀螺剂量,分别在室内和现场进行灭螺试验,观察两种药物的灭螺效果并初步分析评估其成本.结果 在现场气温22～30℃条件下,荣宝50 g/m3浸杀3、5、7 d后,螺袋内钉螺校正死亡率均达到100.0%,与氯硝柳胺2 g/m3灭螺效果相似;荣宝30 g/m2剂量喷洒3、5、7、15 d后,钉螺校正死亡率分别为54.5%、58.0%、69.0%、79.1%,氯硝柳胺喷洒组钉螺校正死亡率分别为61.0%、69.4%、76.7%、77.9%.在室温18℃条件下,荣宝以30 g/m2喷洒3、5、7、15 d后,钉螺校正死亡率分别为72.9%、87.2%、91.5%、76.1%;而相应2 g/m2氯硝柳胺喷洒后的钉螺校正死亡率分别为81.3%、95.7%、97.9%、80.4%.同样完成1000 m2的喷洒灭螺任务,荣宝所需灭螺药物和人力资费成本比氯硝柳胺多支出0.114元/m2;完成72 m3的浸杀灭螺任务,荣宝所需灭螺药物和人力资费成本比氯硝柳胺多支出0.127元/m3.50 g/m3荣宝浸杀灭螺剂量,对成鱼(＞250 g)的活力不会造成影响,但对鱼类幼苗仍具较强毒性.结论 荣宝与氯硝柳胺灭螺效果相似,由于其成本较高,氯硝柳胺仍然是目前首选灭螺药物,但荣宝的鱼类毒性低,可作为氯硝柳胺之外有益的补充灭螺药物.     

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)(.-).