不同实验顺序和不同厂家试剂对测定尿酸的干扰因素分析和解决方案

目的 探讨全自动生化分析仪测定尿酸(UA)时产生交叉污染的来源及避免对策.方法 筛查出对UA测定存在污染的试剂;确定干扰试剂的种类后进行试剂污染试验,检测交叉污染的情况.结果 在试剂干扰检测中发现,高密度脂蛋白胆固醇(HDL-C)测定对UA的测定存在污染;试剂污染试验发现,HDL-C对UA检测结果有明显的正干扰.结论 全自动生化分析仪测定UA会存在项目间的交叉污染,可通过调整项目检测顺序、建立试剂针特殊清洗程序,或者更换具有抗污染成分的试剂等方法避免交叉污染.     

Th17 lymphocyte levels are higher in patients with ruptured than non-ruptured lumbar discs,and are correlated with pain intensity

Background

Th17 lymphocytes have important roles in inflammation and autoimmune disease. Research on relationship between Th17 lymphocytes and pain associated with lumbar disc herniation (LDH) is limited. The purpose of this study was to examine the association of pain and Th17 lymphocyte and interleukin (IL)-17 levels in patients with herniated and non-herniated lumbar discs.

Methods

Thirty-four patients with single lumbar intervertebral disc herniation (median age, 44 years), and 17 healthy adults (median age, 37 years) were enrolled. Patients were divided into 2 groups depending on their magnetic resonance imaging (MRI) results and visual observations during surgery (group P, non-ruptured disc, n = 15; group E, ruptured disc, n = 19). Patients received posterior or transforaminal lumbar interbody fusion. Preoperative pain intensity was recorded using a visual analogue scale (VAS) score. The percentage of Th17 lymphocytes and IL-17 and prostaglandin E2 (PGE2) levels in peripheral blood were determined. Disc tissue was examined by immunohistochemistry for Th17 and IL-17 expression.

Results

Preoperative VAS pain scores were significantly higher in group E than group P (8.32 ± 1.04 vs. 6.33 ± 2.68, respectively, p = 0.009). Similarly, PGE2 level was greater in group E than group P (3.75 ± 1.41 pg/ml vs. 2.63 ± 0.89 pg/ml, respectively, p = 0.011). Compared to healthy controls (1.05 ± 0.19%), the percentage of Th17 cells was significantly greater in group P (1.52 ± 0.62%, p = 0.031), and the percentage in group E (2.99 ± 1.09%, p < 0.001) was significantly greater than in group P. The IL-17 expressions were similar. VAS pain score was positively correlated with Th17 proportion (r = 0.489, p = 0.003), and IL-17 concentration (r = 0.458, p = 0.007). PGE2 was also positively correlated with Th17 proportion (r = 0.539, p = 0.001), and IL-17 concentration (r = 0.500, p = 0.003). The expression of IL-17 was higher in the cells of group E and group P compared with normal tissue (p < 0.001).

Conclusions

Immune system activation is responsible, at least in part, for the pain experienced by patients with LDH, and increased levels of Th17 lymphocytes and IL-17 contribute to the pain.     

生化试剂对循环酶法测定血清总胆汁酸的干扰因素分析和改进措施

目的探讨常规生化试剂对循环酶法测定血清总胆汁酸(Total Bile Acid,TBA)时的干扰因素,寻找消除携带污染的有效方法。方法以TBA为检测项目,以ALT、AST等20个生化常规项目试剂作为观察项目,确认对TBA测定产生明显干扰的常规生化试剂,并通过设置特殊程序减少仪器的携带污染。结果在20个观察项目中,UA试剂对测定TBA结果影响有统计学意义(p<0.01),存在系统误差;TG、CHOL项目试剂对测定TBA结果影响有统计学意义(p<0.05),对TBA测定有明显的正干扰。结论全自动生化分析仪测定TBA时会存在项目间的交叉污染,设置合理的特殊冲洗程序可以有效地消除携带污染对TBA测定的干扰。     

我国体外诊断试剂标准现状分析及思考

目的：为我国体外诊断试剂标准化建设提出相关建议。方法：总结分析我国体外诊断试剂标准化建设及标准体系结构,重点挖掘目前标准化工作与行业发展不适宜的内容,并提出针对性建议。结果与结论：近年来,我国体外诊断试剂行业发展迅猛,国家出台了一系列法规政策,以推动体外诊断试剂行业有序发展。体外诊断试剂在我国分属药品和医疗器械两类,标准体系也分为两部分,相应的标准法规已基本建立健全。但是,目前我国体外诊断试剂标准体系存在标准相对滞后、缺乏强制性标准、一些新兴技术产品尚无标准等问题。需进一步优化体外诊断试剂标准体系,完善标准组织管理并加强标准执行能力,以便更有力地推动我国体外诊断试剂整体质量水平的提升。     

Chemical modification of proteins: comments and perspectives*

The use of chemical modification of proteins has increased exponentially during the past two decades. Today the many different uses of chemical modification include determination of relative reactivities of side chain groups, the quantitation of individual amino acids, development of affinity reagents, mechanism-based reagents for pharmaceutical uses, cross-linking reagents, special techniques for bioprostheses, blocking reagents for peptide synthesis, and reagents for specific cleavages of peptide bonds. Chemical modification should continue to be a primary tool in protein chemistry. It can supply information or products difficult or impossible to attain by the newer powerful technique of in vitro mutagenesis as well as serve as a supplementary procedure for the latter.     

斑点免疫渗滤法检测结核杆菌抗体试剂盒的研制

介绍斑点免疫渗滤法检测结核杆菌抗体（ＴＢ Ａｂ）试剂盒的研制方法。该法是以酶联免疫吸附试验原理和亲和层析原理为基础，采用硝酸纤维素薄膜（ＮＣ膜）作为固相载体而建立一种简便（三步）、快速（５～１０ｍｉｎ）、特异性强、灵敏度高（ｎｇ水平）的新方法。该法的逐步完善将在流行病学调查和临床医学检查中得到广泛应用。     

自配AXSYM全自动免疫分析仪S0lution 3、S0lution 4试剂与原装试剂的比较分析

目的为解决美国雅培AXSYM免疫分析仪Solution 3、Solution 4试剂国产化问题,采用自配试剂以代替原装试剂.方法将自配试剂的理化性质、精密度、准确度与原装试剂进行对比分析.结果自配试剂与原装试剂理化性质一致,采用ME-lA法、FPIA法、ICAI法分别测定AFP、T4、Folate质控液10次,其CV值均小于5%,与原装试剂对比,两者无显著性差异(P＞0.05).结论自配美国雅培AXSYM免疫分析仪Solution 3、Solution 4试剂完全可以代替原装进口试剂.     

Fluorogenic reagents, having benzofurazan structure, in liquid chromatography

Newly synthesized fluorescent reagents, NBD-F and PBD-SO₂F for amines, DBD-F for thiols and amines, and ABD-F and SBD-F for thiols are reviewed in terms of their reactivity, fluorescence characteristics, stability, selectivity, and their applicability to analysis.     

Biologically displayed random peptides as reagents in mapping protein-protein interactions

Summary Random peptide libraries, also known as combinatorial peptide libraries, represent an exciting new set of reagents for the purposes of mapping protein-protein interactions and drug discovery. Millions to trillions of peptides can be expressed biologically, such that the plurality of peptides are accessible and physically linked to the genetic element encoding them. These libraries can be screened by affinity selection to isolate those members in the library that display binding to the target molecule of interest. Sequencing of the associated DNA or RNA elements then leads to the elucidation of the binding peptide's primary structure. From the collection of isolated binding peptides, a consensus motif is frequently evident; these motifs are informative in investigating the interactions of natural proteins. Finally, because synthetic forms of identified peptides are often pharmacological antagonists, they can serve as drug leads.     

Induction of long-term potentiation in the basolateral amygdala does not depend on NMDA receptor activation.

Long-term potentiation (LTP) can be induced in the lateral and basolateral amygdala by stimulating synaptic afferents in the external capsule (EC). We examined the sensitivity of amygdaloid LTP to the NMDA receptor antagonist 2-amino-5-phosphonopentanoate (AP5), which is known to block LTP induction in the Schaffer collateral/CA1 synapses in the hippocampus. While relatively high concentrations (100 microM) of DL-AP5 were effective in preventing LTP induction in the lateral and basolateral amygdala in vitro, the same concentrations also significantly depressed synaptic responses to low-frequency stimulation. Furthermore, at 50 microM, a concentration sufficient to block both synaptic responses mediated by NMDA receptors and LTP induction in the hippocampus and neocortex, AP5 did not affect the probability of inducing LTP in the amygdala. Application of 10 microM 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), which blocks non-NMDA excitatory amino acid receptors, reduced the monosynaptic response to EC stimulation by 85%. The remaining CNQX-insensitive response did not appear to be mediated by NMDA-type receptors, since it was not reduced by 50 or 100 microM AP5, and showed none of the voltage sensitivity characteristic of NMDA responses. These data suggest that while the induction of LTP in the amygdala produced by EC stimulation is blocked by high doses of AP5, plasticity at these synapses probably does not require activation of NMDA receptors.