PCR产物生物发光分析技术研究

目的基于水母发光蛋白生物发光分析技术建立一种PCR产物定量检测技术。方法利用RT-PCR对CK19-mRNA进行放大,PCR引物的5’端用生物素标记,然后用交联了地高辛寡核苷酸探针与目标生物素化的DNA模板杂交。杂交复合物与包被在微孔板上的链亲合素结合,加入标记了水母发光蛋白的地高辛抗体与地高辛特异结合,再加入钙离子激发水母发光蛋白发光,发光强度正比于CK19-mRNA的量。结果探测灵敏度可达22 amol/L的RT-PCR产物。结论该定量检测技术是一种灵敏的、可靠的、非放射性的微量CK19-mRNA定量方法。     

化学发光法检测血清CK18水平在胃癌诊断中的应用

目的 寻找新型血清CK18的检测方法,并作为一种胃癌肿瘤标志物与CA72-4指标进行比较.方法 分别以CK-3A9和CK-2A6作为固相抗体和标记抗体,制备化学发光试剂盒(CK18-3A9)对正常人及胃癌患者血清CK18进行检测;通过ROC曲线比较CK18-3A9与CA72-4在胃癌诊断中的意义.结果 CK18-3A9化学发光测定试剂盒诊断胃癌灵敏度为44.00%,特异度为98.50%;诊断胃腺癌灵敏度为42.65%,特异度为98.50%;诊断正常人血清假阳性率为0%;诊断炎症血清假阳性率为6.00%.结论 在该研究考察的范围内,与CA72-4相比,CK18-3A9在诊断胃癌时有更好的灵敏度和特异度.     

CTn-I与CK、CK-MB检测在急性心肌梗死中的应用比较

目的评价肌钙蛋白（cardiac troponin—I，CTn—I）与肌酸激酶（CK）、肌酸激酶同工酶MB（creatine kinase isoenzyme MB，CK—MB）等检测指标对急性心肌梗死的诊断的临床准确性。方法对近二年所测定的150例患者的心肌酶谱及肌钙蛋白（CTn—I）和临床诊断治疗的资料做了回顾性分析。结果通过以上分析，我们可以看出CTn—I检测兼有心肌酶的优点，同时比心肌酶特异性更高，窗口期更长、阳性结果表现更早，金标法CTn—I检测卡操作更为方便，尤其适合急诊床边检测的需要。以往通过心电图和心肌酶进行确诊时容易出现漏诊或操作繁琐，往往延误了治疗的最佳时机，而CTn—I检测卡可以用全血在15min之内快速诊断，可以在第一时间为临床治疗提供最为准确的诊断依据。结论CTn—I能提高诊断的准确率和减少漏诊。     

高压氧对正常大鼠血中一氧化氮及磷酸肌酸激酶含量的影响

目的　探讨 0 .2 MPa高压氧 (HBO)对正常组织一氧化氮 (NO)系统和血清磷酸肌酸激酶含量的影响。方法　实验用 Wistar大鼠 36只 ,随机分成对照组和 HBO处理后第 1、5、1 0、2 0、30天组。分别于 0 .2 MPa HBO处理后第 1、5、1 0、2 0、30天用流式细胞仪测定大鼠海马钙离子 ([Ca2 ]i)水平、用分光光度法测定血浆 NO浓度、用放免法测定大鼠海马神经原环磷酸鸟苷 (c GMP)含量及用琼脂糖凝胶电泳法测定血清磷酸肌酸激酶脑型同功酶 (CK- BB)浓度的变化。结果　 HBO处理后第 1、5、1 0、2 0天 NO系统与对照组相比虽有所变化 ,但差异无显著性 (P>0 .0 5 )。血清 CK- BB在 HBO的第 1、5、1 0、2 0天与对照组相比差异亦不显著 (P>0 .0 5 ) ,而第 30天组 ,除 c GMP外 ,其它指标均明显异常 (P<0 .0 5 )。结论　 0 .2 MPa HBO环境暴露每日 1次持续 2 0 d,未见其引起 NO系统损伤及脑损伤所致的血清酶学的改变。说明在一定时程内和一定压力下 ,HBO处理对正常组织是安全的     

Indications of hepatic and cardiac toxicity caused by subchronic Tricholoma flavovirens consumption.

The yellow tricholoma (Tricholoma flavovirens or Tricholoma equestre) is a wild mushroom species that was previously considered edible and tasty. Recently, it caused several cases of delayed rhabdomyolysis in humans and elevated serum creatine kinase (CK) activities in laboratory mice (Mus musculus) in a dose-response study. The present study continued to examine the effects of prolonged T. flavovirens consumption at 12 g freshly frozen mushroom kg(-1)d(-1) on the plasma clinical chemistry and organ histology of mice. The plasma CK and CK-MB activities and the plasma bilirubin concentrations were higher in the exposed mice than in the controls. In addition, pericardial lymphocyte infiltrates were present in the mice that had consumed the mushroom. The results indicate myo-, cardio- and hepatotoxic effects of T. flavovirens.     

New responsibilities for aged kinases in B-lymphomas

The knowledge accumulated over the last decade on B-cell–derived non-Hodgkin lymphoma (B-NHL) pathogenesis has led to the identification of several molecular abnormalities, opening new perspectives in the design of novel therapies. Indeed, drugs targeting specific biochemical pathways critical for B-NHL cell survival, proliferation, and fitness within the malignant microenvironment are now available to the clinician: the B-cell receptor signaling inhibitors of BTK, PI3Kδ, ζ, γ, and SYK or the pro-apoptotic BH3-mimetics are clear examples of it. Moreover, it is emerging that malignant B-cell growth is sustained not only by mutations in oncogenes/tumor suppressors but also by the “addiction” to nononcogene (ie, nonstructurally altered) molecules. In this regard, a consistent body of data has established that the Ser/Thr kinases CK1, CK2, and GSK3 are involved in malignant lymphocyte biology and act as pro-survival and signaling-boosting molecules, both in precursor and mature B-cell tumors. Currently, an experimental and clinical groundwork is available, upon which to design CK1-, CK2-, and GSK3-directed antilymphoma/leukemia therapies. In this review, we have examined the main features of CK1, CK2, and GSK3 kinases, summarized the data in B-NHL supporting them as suitable therapeutic targets, and proposed a perspective on potential future research development.     

CK19和DNA拓扑异构酶Ⅱ在甲状腺肿瘤患者组织中表达的临床意义

目的 研究甲状腺肿瘤患者组织中CK19和DNA拓扑异构酶Ⅱ与甲状腺肿瘤病理特征的相关性.方法 用免疫组织化学法检测23例甲状腺肿瘤患者组织中CK19和DNA拓扑异构酶Ⅱ的轰达.结果 12例甲状腺癌患者组织中CK19均阳性表达,DNA拓扑异构酶Ⅱ阴性表达;而在11例良性甲状腺病变患者组织中DNA拓扑异构酶Ⅱ阳性表达,CK19呈阴性表达.结论 CK19可以作为甲状腺癌的肿瘤标志物之一;而对于TopoⅡα呈阳性表达的良性甲状腺疾病,可以选择对该酶具有敏感性的药物进行治疗,以及明确诊断提供依据.