南京市两对半中标试剂与美国Abbott试剂的比较

目的：对南京市2004年中标的五家公司的两对半试剂（ELISA法）进行评价。方法：利用美国Abbott作为参比试剂,对五家中标试剂分别进行对比、评价。结果：五家公司中标试剂与美国Abbott试剂相比特异性较好但敏感性存在一定差异,五家中标试剂之间也存在质量上的差别。结论：五家中标试剂可以使用但质量均有待提高。     

Quantitative immunohistochemical expression of c Kit in breast carcinomas is predictive of patients' outcome

Background:

c Kit (CD117) expression in tissues has been reported as a relevant target for specific therapy in some human malignancies, but has been poorly documented in breast carcinomas

Methods:

The prognostic significance of c Kit in a series of 924 breast carcinomas (mean follow-up, 79 months) was investigated using standardised high-throughput quantitative densitometry of immunohistochemical precipitates in tissue microarrays.

Results:

c Kit was expressed in 14.7% breast carcinomas (and in 42 out of 586 node-negative tumours). In univariate analysis, (log-rank test) the score of c Kit expression correlated with poor patient outcome P=0.02 and particularly in node-negative cases (P=0.002). In multivariate Cox analysis, c Kit was an indicator of metastasis independent of 25 other concomitantly evaluated markers of prognosis. Logistic regression showed that c Kit ranked 10 out of 25 (P=0.041), and was included in a 10-marker signature that allowed 79.2% of the patients to be correctly classified in the metastatic or metastasis-free categories independently of hormone receptors and HER-2 status. Interestingly, c Kit was also a significant predictor of metastasis in node-negative tumours (2 out of 25 ranking, P<0.0001) and included in a six-marker signature of prognosis, correctly classifying 88.6% of the patients (P<0.0001).

Conclusion:

We concluded that, as assessed by quantitative immunohistochemistry, c Kit is an independent prognostic indicator that could also potentially serve as a target for specific therapy in breast carcinomas.     

Fine-needle aspiration cytology diagnosis of metastatic gastrointestinal stromal tumor in the liver: a report of three cases

Gastrointestinal stromal tumor (GIST) is the designation for a major subset of gastrointestinal mesenchymal tumors that histologically, immunocytochemically, and genetically differ from leiomyomas, leiomyosarcomas, and schwannomas. GISTs derive from the interstitial cells of Cajal and, in addition to variable expression of smooth muscle and neural markers, they characteristically express CD34 and CD117. The cytological appearance, including immunocytochemical and mutational analysis of c-kit gene in primary GIST has been well described. To our knowledge, only two cases of metastatic GIST diagnosed by fine-needle aspiration (FNA) have been reported. We illustrate three cases of metastatic GIST in the liver. Two cases had no prior history of gastrointestinal tumor and the third case had a 4-yr previous history of duodenal tumor. Consistent immunocytochemistry and ultrastructual studies supported the diagnosis of GIST. We emphasize that in the appropriate clinical and radiological setting, a confident diagnosis of GIST can be established by FNA of metastatic lesions.     

动物组织中盐酸克伦特罗残留ELISA检测试剂盒的研制

目的本研究采用直接竞争酶联免疫吸附试验(ELISA)对动物组织中盐酸克伦特罗(CBL)残留量进行快速筛选检测。方法将CBL与蛋白质载体偶联制备了CBL-BSA和HRP-CBL结合物,以CBL-BSA免疫实验兔获得特异性抗体,并成功建立了CBL残留直接竞争ELISA检测方法及检测试剂盒。结果检测线性范围0.1-62.5 ng/mL,检测低限为0.10 ng/mL;尿、肝、肉和饲料的加标回收率为60.3%-115.0%;与英国Randox试剂盒对比测试结果无显著差异;重现试验变异系数为3.4%-7.2%;试剂盒贮藏在2-8℃5个月和30℃7 d质量稳定。结论本试剂盒具有操作简便、快速、灵敏等特点。     

两种提取人参水煎液miRNA方法的比较

目的 筛选获取新鲜人参水煎液中高质量miRNA的提取方法.方法 选择改良Trizol法和通用植物miRNA提取试剂盒法提取新鲜人参水煎液miRNA,所得miRNA经DNaseⅠ处理后进行琼脂糖凝胶电泳和Agilent 2100生物分析仪检测,比较两种方法所得miRNA的纯度和质量.结果 改良Trizol法步骤简单、快捷省时、成功率较高但纯度较低;通用植物miRNA提取试剂盒法步骤较多耗时较长,但miRNA纯度高质量好.结论 通用植物miRNA提取试剂盒法更适用于提取人参水煎液中miRNA,提取的miRNA完整性好、纯度高,可以满足荧光定量PCR、建库测序等后续研究的需要.     

输卵管介入再通单人操作套件与双人操作套件的对比研究

目的:探究输卵管介入再通(FTR)手术单人操作的可行性,提出单人操作套件的改进方案。方法:选取经子宫输卵管造影(HSG)诊断的65例输卵管近端梗阻患者,根据FTR术中操作方法不同,将采用双人套件操作的患者纳入双人套件组(33例),采用单人套件操作的患者纳入单人套件组(32例),比较两组患者FTR术中子宫形态及位置、钳夹宫颈与否、再通术成功率、透视时间、辐射剂量及患者视觉疼痛量表(VAS)得分情况。结果:两组FTR术中再通成功率及相关手术指标比较,差异均无统计学意义(t=1.34,t=1.12,t=0.89,t=0.56;P>0.05);三角形宫腔患者手术透视时间长、术后VAS得分高,与倒三角形及梭形宫腔比,三角形宫腔患者差异有统计学意义(F=6.34,F=5.22;P<0.05)。结论:FTR术中单人操作套件可较好地替代双人操作套件功能,对于操作难度高的患者,单人操作套件仍需改进;在三角形宫腔操作困难时应尽早采用双人操作。     

可溶性人胎盘生长因子ELISA检测试剂盒的建立及性能分析

目的:研发人血清及尿液可溶性胎盘生长因子(sPLGF)的ELISA检测试剂盒并进行性能测试。方法:以抗人PLGF单克隆抗体包被微孔板,加入重组sPLGF和生物素标记的检测抗体,形成抗原抗体复合体,加入链亲合素辣根过氧化物酶和底物TMB实现比色反应;从线性范围、准确度、精密度、样品基质及样品前处理方法等指标进行性能评价,并与市售进口同类产品进行比较。结果:捕获抗体的最适包被量为400ng/well,包被条件为4℃孵育16~20h,检测抗体的最适工作浓度为1μg/ml,样品最适反应条件为37℃孵育2h,适合的封闭液及样品稀释液组合为3%BSAPBS和0.1%BSA-PBS;验证该试剂盒的有效检测范围为15.6~2000pg/ml,准确度为85%~115%;批次内变异系数CV≤10%,批次间变异系数CV≤15%;其检测范围、准确度、精密度、灵敏度及样品适用范围等与市售进口试剂盒基本一致。结论:本项目开发的人sPLGF检测试剂盒实现了人血清和尿液中的内源sPLGF的定量检测,其检测性能稳定、可靠,具有比进口试剂盒检测范围更广、价格更低的优势,可用于临床检测血液及尿液中的内源性sPLGF水平。