HLA-DRB1、-DQB1基因多态性与食管鳞癌遗传关联性

目的　从基因水平探讨食管鳞癌HLA DRB1 , DQB1等位基因的遗传易感性 ,以阐述其免疫遗传学特征。方法　运用序列特异性引物聚合酶链反应技术 ,检测无亲缘关系湖北汉族健康人 1 36例、食管鳞癌患者 42例的HLA DRB1 , DQB1等位基因。结果　湖北汉族人食管鳞癌患者与正常人比较 ,HLA DRB1 0 90 1等位基因分布频率显著增高 (0 .2 50 0比 0 .1 397,P =0 .0 2 8,OR =2 .0 53 ,病因分数 =0 .1 2 82 ) ,HLA DQB1 0 30 1基因分布频率显著增高 (0 .2 976比 0 .1 875 ,P =0 .0 4 6 ,OR =1 .835 ,病因分数 =0 .1 35 4)。两者间其余HLA DRB1、 DQB1等位基因分布频率差异均无显著性。结论　HLA DRB1 0 90 1及 DQB1 0 30 1等位基因均与食管鳞癌正关联 ,为其易感基因。该两等位基因测序结果与其基因库第 2外显子序列吻合。     

HLA-DQB1-HLA-DRB1单倍型与中国南方汉族肺结核的相关性分析

目的　探讨HLA DQB1 HLA DRB1单倍型在中国南方汉族肺结核发病机制中的可能作用。方法 采用病例 对照的研究方法 ,应用PCR SSP技术对110例中国南方汉族肺结核患者和 10 1例中国南方汉族健康对照者的 2 0个HLA DRB1和 8个HLA DQB1等位基因进行分型 ,比较两组间DQ2,3(8) DRB1、DQ3(7) DRB1、DQ3(8,9) DRB1、DQ2 ,3(7,9) DRB1、DQ2 DRB1、DQ4 DRB1、DQ5DRB1和DQ6 DRB1单倍型频率 (HF)并计算其相对危险性 (RR)。结果 DQ2 ,3(8) DR14 .1、DQ3(7)DR16单倍型的频率肺结核病例组显著高于对照组 (6 .10vs .0 .5 0、4 .18vs .0 .99) ,其RR分别为 13.4 0和 4 .41;DQ2 DR1、DQ2 DR12、DQ2 DR13.3、DQ3(7) DR1、DQ3(7) DR13.3、DQ3(8,9) DR13.3、DQ2,3(7,9) DR1、DQ2 ,3(7,9) DR13.3、DQ2 ,3(7,9) DR13.4、DQ4 DR4单倍型的频率肺结核病例组显著低于对照组(分别为 1.84vs .5 .6 0、1.37vs .5 .6 0、4 .18vs .11.0 0、2 .30vs .9.89、12 .6 2vs .2 2.2 8、5 .6 1vs .11.5 6、3.70vs .14 .4 0、16 .88vs .2 8.94、5 .13vs .12 .12、2 .30vs.6 .13) ,其RR分别为 0 .31、0 .2 3、0 .34、0 .2 1、0 .4 7、0 .4 4、0 .4 6、0 .38和 0.35。结论 DQ2 ,3(8) DR14 .1、DQ2 DR12、DQ2 ,3(7,9 )-DR13.4、DQ4-DR4单倍型的存在可能与中国南方汉族肺结核的发病有着关联,而其他单倍型差异的显著性则可能是因其组成基因的基因频率差异所致。     

HLA-DR基因与中国南方汉族部分人群肺结核易感基因的研究

目的　探讨人类白细胞抗原 (HLA) DR基因与中国南方汉族部分人群肺结核发病的关联性 ,并寻找与肺结核发病可能相关的HLA易感基因。方法　采用病例 对照的研究方法 ,应用聚合酶链反应 序列特异性引物 (PCR SSP)技术对 110例中国南方汉族肺结核患者 (肺结核病例组 )和 10 1例中国南方汉族健康对照者 (健康对照组 )的 2 3个HLA DR等位基因进行分型 ,比较其等位基因频率(GF)并计算其比值比 (OR)。结果　HLA DR基因PCR SSP分型显示 :(1)肺结核病例组中的DR16等位基因的基因频率显著高于健康对照组 ,两组的GF值分别为 12 6 2 %、5 6 0 % ,两者之间差异具有显著性 (χ2 =5 915 ,PC<0 0 5 ,OR值为 2 5 3)。 (2 )肺结核病例组中的DR1、DR13 3等位基因的基因频率分别为 8 0 8%、2 3 5 7% ,显著低于健康对照组的 2 9 2 9%、5 0 2 4 % ,两组比较 ,差异具有显著性 (χ2值分别为 17 84 7和 14 2 5 8,PC 值均 <0 0 1;OR值分别为 0 2 6、0 33)。结论　 (1)DR16等位基因与南方汉族部分人群的肺结核发病可能密切相关 ,或与真正起作用的易感基因连锁。 (2 )中国南方汉族部分人群DR1、DR13 3等位基因的表达对结核分枝杆菌感染者的发病可能具有拮抗作用。     

HLA-DRB1、DQB1基因与汉族哮喘的相关性研究

探讨 HL A- DRB1、DQB1位点基因在中国汉族哮喘家系中与哮喘的相关性。用序列特异性引物 -聚合酶链反应 (PCR- SSP)方法 ,对 10 1例哮喘家系成员和 6 0例正常对照者进行了 HL A- DRB1、DQB1等位基因的分型 ,并分析了 DRB1、DQB1基因在两组中的分布。结果示 ,与正常对照组比较 ,哮喘患者组 DRB1* 15等位基因频率 (2 0 .93% )较正常对照组 (6 .6 7% )明显增高 (χ2 =10 .95 ,P<0 .0 5 ) ;DQB1* 0 6 0 1等位基因频率在哮喘患者组(31.4 0 % )较正常对照组 (7.5 % )明显增高 (χ2 =2 3.0 8,Pc<0 .0 1)。同时发现 HL A- DRB1* 0 7等位基因频率在对屋尘螨抗原皮试阳性家系成员中 (2 7.4 2 % )较家系中皮试阴性者 (5 .36 % )显著增高 (χ2 =10 .83,Pc<0 .0 5 )。HL A- DRB1* 15和 DQB1* 0 6 0 1可能是汉族哮喘的遗传等位易感基因 ,HL A- DRB1* 0 7在限定对屋尘螨抗原特异性 Ig E反应过程中起重要作用     

北京地区中老年人糖尿病和IGT患病率与增龄相关性的分析

目的　阐明中老年人糖尿病 (DM)和 IGT患病的增龄性特点及增龄的患病风险。方法　采用随机分级整群抽样方法 ,横断面调查北京市城乡 4个社区 2 35 4例≥ 4 0岁的中老年人。结果　 DM患病率为 11.36 % (标化率 :10 .6 4 % ) ,IGT患病率为 12 .17% (标化率 :11.5 6 % ) ,其与增龄有显著的相关性 (DM:r =0 .99,P =0 .0 0 2 ;IGT:r =0 .92 ,P =0 .0 2 9)。不同年龄组的血糖均值秩和分析 (IGT:χ2 =9.816 ,P =0 .0 4 4 ,总体 :χ2 =10 6 .94 4 ,P <0 .0 0 0 5 )在IGT和总体中证明有显著随年龄增加而增长的趋势。有 3个显著不同的患病风险 (OR)年龄阶段 ,即 4 0～ 4 9岁最低(DM:4 .95 % ,OR=0 .4 1;IGT:8.35 % ,OR=0 .6 6 ) ;5 0～ 6 9岁较高 (DM:12 .13%～ 13.5 7% ,OR=1.0 8～ 1.2 2 ;IGT:11.0 2 %～ 13.6 0 % ,OR=0 .89～ 1.13) ;70～ 99岁最高 (DM:19.0 2 %～ 2 1.0 5 % ,OR=1.83～ 2 .0 8;IGT:19.0 1%～2 1.0 5 % ,OR=1.6 9～ 1.92 )。患病高峰年龄为 DM(95 %可信区间 ,CI) :4 1.0～ 80 .4岁 ,IGT(95 % CI) :36 .9～ 82 .1岁。结论　因 DM和 IGT患者人数随增龄积累 ,年龄越大 ,患病率越高 ,表现出高龄群体高患病风险的增龄特点。年龄是 DM发生的一独立危险因素。     

山东地区类风湿关节炎与HLA-DRB1基因共同表位的关联性研究

目的　探讨山东地区汉族人群类风湿关节炎 (RA)与HLA DRB1基因共同表位 (SE)的关联性。方法　采用特异性引物聚合酶链反应 (PCR SSP)方法对山东地区人群 1 32例RA患者及1 30名正常健康者的HLA DRB1 0 1、 0 4、 1 0的 1 7个等位基因进行检测。结果　山东地区RA患者中携带有SE的基因频率显著高于正常对照组 (5 0 0 %∶2 2 3% ,P <0 0 1 ) ,HLA DR4亚型 0 4 0 5是主要的易感基因 (2 2 8%∶1 0 0 % ,P <0 0 0 5 )。其他亚型包括DRB1 0 1 0 1 (3 8%∶3 1 % ) , 0 1 0 2 (2 3%∶2 3% ) , 0 1 0 3(3 8%∶3 1 % ) , 0 1 0 4 (3 0 %∶2 3% ) , 0 4 0 1 (1 0 6 %∶4 6 % ) , 0 4 0 4 (9 1 %∶4 6 % ) , 0 4 0 7(8 3%∶9 2 % ) , 0 4 0 3(6 8%∶3 1 % ) , 0 4 0 2 (6 8%∶4 6 % ) , 0 4 0 8(5 3%∶1 5 % ) , 0 4 0 9(2 3%∶0 ) , 0 4 0 6 (1 5 %∶0 ) , 0 4 1 0 (0 8%∶0 ) , 0 4 1 1 (0 8%∶0 )和 1 0 0 1 (1 1 4 %∶6 9% )的差别均无统计学意义。Logistic回归分析表明 :SE纯合子对RA的危害性要比其杂合子大 (P <0 0 0 1 )。结论　SE与山东地区汉族RA易感性及疾病严重性有关联     

慢性乙型肝炎干扰素疗效与人类白细胞抗原-等位基因的关系

目的　探讨慢性乙型病毒性肝炎干扰素 α治疗 6个月疗效与人类白细胞抗原 (HLA)部分等位基因的关系。方法　应用聚合酶链反应 序列特异性引物 (PCR SSP)技术对 6 0例上海地区接受干扰素 α正规治疗 6个月的慢性乙型肝炎患者的HLA Ⅱ类分子DRB1、DQA1、DQB1进行等位基因多态性分析。结果　无应答组的HLA DRB1 0 4携带率高于有应答组 (P <0 .0 2 5 ) ,有应答组的HLA DQA1 0 5 0 5 (P <0 .0 2 5 )、DQB1 0 30 1(P <0 .0 0 5 )的携带率高于无应答组。结论　干扰素 α治疗无应答与HLA DRB1 0 4 (P <0 .0 2 5 )呈正相关 ,与HLA DQA1 0 5 0 5、HLA DQB1 0 30 1呈负相关     

细胞间黏附分子-1基因K469E多态性与冠状动脉支架置入后再狭窄相关性的初步探讨

目的　炎症反应是冠状动脉支架置入后再狭窄的重要因素 ,细胞间黏附分子 1(ICAM 1)是介导白细胞在血管内皮细胞表面紧密黏附的重要黏附分子。本研究的目的在于探索ICAM 1基因多态性与中国北方汉族人群中冠状动脉支架置入后再狭窄的相关性。方法　收集冠状动脉支架术后行冠状动脉造影随访的患者 12 4例 ,同时收集传统危险因素、手术相关因素信息。应用PCR RFLP方法确定ICAM 1K4 6 9E基因型。结果　入选的 12 4例患者中再狭窄者 72例 ,无再狭窄者 5 2例 ;12 4例中ICAM 1K4 6 9E基因型频率为 :KK纯合子 5 0 8% ,EE纯合子 7 3% ,EK杂合子 4 1 9%。再狭窄患者中KK纯合子和E等位基因携带基因型的频率分别为 5 8 3%、4 1 7% ;无再狭窄患者中二者的频率分别为 4 0 4 %、5 9 6 %。二者分布差异有显著性 (P =0 0 4 9)。调整混杂因素后显著性差异更为明显 ,KK纯合子OR值为 2 6 ,95 %可信区间为 1 2～ 5 8(P =0 0 18)。危险因素分层发现在肥胖 (体重指数≥ 2 7)、高脂血症患者KK纯合子的再狭窄危险更高 ,OR值分别为 9 3、3 7(P值均小于 0 0 5 )。结论　ICAM 14 6 9KK纯合子冠状动脉支架置入后再狭窄危险性较高 ,且在肥胖或高脂血症患者中作用更为显著。     

新疆地区维吾尔族及汉族溃疡性结肠炎患者人类白细胞抗原-DRB1等位基因与抗中性粒细胞胞质抗体的相关性

目的 研究人类白细胞抗原(HLA)-DRB1基因多态性与新疆地区维吾尔族及汉族溃疡性结肠炎(UC)患者抗中性粒细胞胞质抗体(ANCA)的关联性.方法 用间接免疫荧光法分别对62例维吾尔族UC患者、58例汉族UC患者、188名维吾尔族健康对照者、184名汉族健康对照者进行血清ANCA检测,采用聚合酶链反应直接测序分型法(PCR-SBT)进行HLA DRB1基因分型,分别在维吾尔族和汉族内比较ANCA阳性者、ANCA阴性者及健康对照者的HLA-DRB1等位基因频率,并按UC临床类型、严重程度、受累范围进行分层分析.应用SPSS 17.0统计软件进行x2检验,当P＜0.05时,计算比值比(OR)和95％可信区间(95％ CI).结果 维吾尔族UC患者ANCA阳性率[53.2％ (33/62)]高于汉族UC患者[34.5％ (20/58)],差异有统计学意义(x2=4.269,P＝0.045).在维吾尔族中,ANCA阳性UC患者HLA-DRB1* 13的基因频率(0.202)显著高于ANCA阴性患者(0.017,x2＝10.092,P＝0.016,OR＝16.000,95％CI:2.892～88.524)和健康对照组(0.075,x2=9.351,P＝0.040,OR=3.407,95％CI:1.666～6.971).ANCA阳性的全结肠炎型UC患者HLA-DRB1* 13基因频率(9/15)高于ANCA阴性的全结肠炎型UC患者(1/14),差异有统计学意义(x2=8.955,P=0.040,OR＝19.500,95％CI:2.787～136.461).在汉族中,HLA-DRB1 各等位基因在ANCA阳性者、ANCA阴性者和健康对照者间差异均无统计学意义(P均＞0.05),分层分析所获结果亦然.结论 在新疆地区维吾尔族UC患者中,HLA-DRB1* 13可能与ANCA有关,还可能与全结肠炎型患者的ANCA有关.在新疆地区汉族患者中则可能无此关联性.     

HLA-DR-DQ单倍型与类风湿关节炎的关系

目的　探讨HLA -DR -DQ单倍型与我国汉族类风湿关节炎 (RA)的相关性。方法　以 10 0名健康人为对照 ,采用PCR -RFLP法对汉族人群中 35例RA患者的DRBl、DRB3、DRB5、DQAl和DQBl基因位点多态性进行分析。结果　DRBl 0 4 0 5 -DQAl 0 30 1-DQBl 0 4 0 1单倍型频率在RA患者明显高于正常人 (分别为14 0 %和 4 5 % ,P <0 0 1) ,该单倍型阳性RA患者的病情重于其它RA患者 ,包括关节肿胀数、晨僵时间、RF滴度和病情严重例数在阳性组明显高于阴性组 (P均 <0 0 5 ) ;而DRBl 15 0 1-DRB5 0 10 1-DQAl 0 10 2 -DQBl 0 6 0 2单倍型频率在正常人明显高于RA患者 (分别为 12 %和 4 3% ,P <0 0 1)。结论　DRBl 0 4 0 5 -DQAl 0 30 1-DQBl 0 4 0 1单倍型与RA发病及病情严重程度相关 ;而DRB1 15 0 1-DRB5 0 10 1-DQA1 0 10 2 -DQBl 0 6 0 2单倍型则可能对易患RA起保护作用     

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.     

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.     

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.     

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)(.-).     

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.