基于磁珠的ABO血型蛋白质芯片的构建及初步应用分析

目的：构建一种以磁珠为标记物的,用于血型检测的可视化蛋白质芯片。方法用人免疫球蛋白G （IgG）及磁珠标记的抗人IgG进行琼脂糖浓度、点样浓度、温度和固定时间等实验条件的摸索。根据优化的条件,经过铺片、点样、固定、封闭等程序构建ABO血型检测蛋白质微阵列,加入血液样品进行反应,最后用磁珠标记的抗体进行检测。结果通过条件的优化发现,在1．0％的琼脂糖基片表面,以100μg/m L的蛋白质稀释液点样,之后在24℃固定8 h ,能够得到更优的信号强度。检测不同稀释度的抗原抗体,正定型效价可达到1∶8192,反定型可达到1∶4096。检测14例临床样品,结果样点清晰、肉眼可见,且结果判定与试管法一致性达100％。结论成功构建了基于磁珠的血型检测可视化蛋白质微阵列。     

布鲁氏菌血清 IgM,IgG 抗体胶体金免疫层析检测方法的建立及初步临床应用

目的　应用胶体金免疫层析技术建立一种快速检测布鲁氏菌 IgM,IgG 抗体的方法,并进行初步的性能验证。方法　采用枸椽酸三钠还原法制备胶体金颗粒,标记纯化的抗重组布鲁氏菌抗原,将鼠抗人 IgG 抗体、鼠抗人 IgM 抗体和兔抗布鲁氏菌抗体喷在硝酸纤维素膜上作为检测线和质控线,对 pH 值和抗体浓度等条件优化;评估试剂盒灵敏度、特异度、交叉反应、准确度、精密度和干扰实验;用该方法对临床的 400 份血清进行检测,试管凝集试验验证其准确性。结果　该试纸条最佳胶体金标记的 pH 值为 7.5,标记重组抗原质量浓度为 30?g/ml。方法的灵敏度和稳定性较好且与其他病原菌无交叉反应。血清总胆红素、血脂和血红蛋白浓度的增高会使试纸条显色结果减弱。对400份临床样本进行检测,其结果与试管凝集试验的符合率为 93.75%。结论　该试验建立的布鲁氏菌 IgM/IgG 抗体的胶体金免疫层析检测方法具有较好的特异度和稳定性,整个检测过程可在 15min 内完成,能够快速检测样品血清,适用于现场快速检测。     

软海绵酸胶体金免疫检测法的建立与应用

目的 建立并应用软海绵酸(OA)胶体金免疫层析检测法.方法 通过纯化抗OA单抗、制备胶体金、标记单抗、制备胶体金试剂条、样品模拟提取与加标回收检测、贝类染毒提取及检测,建立和优化金标免疫层析法.结果 选择30 nm胶体金颗粒用于单抗标记,金标单抗浓度为200 μg/mL,金标单抗包被量为2.5 μL/cm,二抗包被浓度为1.0 mg/mL,反应时间为3～5 min,对加标贝肉和染毒贝类的检测灵敏度为25 ng/mL.结论 建立了可用于OA检测的胶体金免疫层析检测法,满足规定的贝类OA含量安全阈值,为腹泻性贝毒检测与食品安全检验提供了新的技术方法.     

胶体金免疫斑点法检测人巨细胞病毒pp150抗体

目的：建立一种简便、快速的检测血清中人巨细胞病毒（HCMV）抗pp150抗体的胶体金免疫斑点法。方法：采用Frens法制备胶体金颗粒，标记葡萄球菌A蛋白（SPA），将本所利用基因工程技术生产的pp150抗原固定于0．45μm孔径的硝酸纤维素膜上，制成免疫斑点检测装置。血清中抗pp150 IgG与硝酸纤维素膜上的抗原结合，然后滴加胶体金标记的SPA，在膜上可形成肉眼可见的红色斑点。结果：胶体金免疫斑点法与ELISA试剂盒对比检测了200份临床确诊血清标本中抗pp150 IgG抗体，胶体金法的特异性和敏感性分别为92％和90％。结论：胶体金免疫斑点法检测血清中的抗pp150抗体特异性、灵敏度较高，简便快速，不需要昂贵的仪器。     

温育加稀释法可解除冷凝集素对血常规检测的干扰

摘要：目的：解决冷凝集素对血常规检测的干扰。 方法：分别在室温（25 ℃）、37 ℃温育15 min（温育法）和37 ℃温育15 min后1∶2稀释（温育加稀释法）3种不同条件下,用Sysmex-2100血液分析仪检测健康人对照组及冷凝集素阳性组的EDTA-K²抗凝全血标本的血常规。 结果：对照组温育法的血常规检测结果与温育加稀释法无明显差异,而冷凝集素阳性组温育加稀释法检测结果优于温育法。 结论：温育加稀释法可以有效地解决冷凝集素对血常规检测的干扰。     

斑点金免疫渗滤法检测大肠杆菌的研究

目的建立一种快速、简便、准确、灵敏度高的大肠杆菌斑点金免疫渗滤检测方法。方法选用混合纤维素膜为固相载体,以胶体金标记羊抗兔IgG,双抗免疫渗滤检测大肠杆菌。结果建立了灵敏度为253~1.04×106个/ml的简便、快速的大肠杆菌检测方法。结论斑点金免疫渗滤实验检测大肠杆菌与传统的检测大肠杆菌方法比较,具有检测速度快、灵敏度高、应用方便等优势。     

等位基因特异性PCR结合金磁微粒免疫层析检测膀胱癌患者尿液中TERT突变

目的建立可检测人尿液DNA中端粒酶逆转录酶(TERT)启动子区域突变的方法。方法设计等位基因特异性PCR引物,利用温度梯度PCR确定等位基因特异性PCR程序,利用金磁微粒检测板进行扩增产物检测。收集34例膀胱癌患者的尿液,提取DNA,利用该文建立的等位基因特异性PCR结合金磁微粒免疫层析方法检测TERT启动子区域突变。结果建立了结合等位基因特异性PCR和金磁微粒免疫层析的TERT启动子区域C228T和C250T突变检测方法。34例膀胱癌患者尿液中8例患者存在C228T突变,1例患者存在C250T突变。结论该研究建立了TERT启动子区域C228T和C250T突变检测方法,可应用于膀胱癌患者尿液DNA突变检测。     

检测肌红蛋白的半定量金免疫层析法的研究

目的建立一种半定量检测人血清肌红蛋白的金免疫层析方法,用于急性心肌梗死的快速检测。方法一株抗人肌红蛋白单克隆抗体标记胶体金,另一株抗人肌红蛋白单克隆抗体包被膜作为检测线,兔抗鼠IgG抗体包被膜作为对照线,建立半定量金免疫层析测定方法。结果本法测定结果分为3个半定量判定区段：〈70、70-150和〉150μg/L;与化学发光免疫测定对照检测83份血清标本,符合性良好。结论金免疫层析测定法具有半定量的特点,方法简便、快速、准确,适用于急性心肌梗死的早期检测。     

免疫金银染色法在蛋白质微阵列检测TORCH感染的实验研究

目的研制基于免疫金银染色法的TORCH检测蛋白质微阵列。方法将TORCH抗原点样于已经被琼脂糖膜修饰的玻片表面,与待测血清和胶体金标记的抗人IgM(μ链)孵育,用银增强微阵列信号,肉眼观察或普通的平板扫描仪记录结果。以不同浓度的纯化人IgM制备标准曲线。结果蛋白质微阵列的灰度质和IgM的浓度呈线性关系。60份待测血清与ELISA法同时检测,诊断符合率为93.3%。结论该法具有高信息通量、低成本、制备与检测快速简单的优点。     

滴金法同步检测甲、乙、丙型肝炎病毒免疫标志物方法学建立及评价

目的　建立同步检测甲、乙、丙型肝炎病毒免疫标志物的金免疫渗滤法 (GIFA)的实验方法 ,并对其性能作评价。方法　采用胶体金作为免疫示踪物 ,标记羊抗人IgG单克隆抗体 ,将相应的病毒抗原HAV Ag、HBc Ag及HCV Ag包被在硝酸纤维素薄膜表面 ,标本中所含抗体与相应的抗原结合后 ,加入金标记抗体 ,即可在薄膜表面形成肉眼可见的红色斑点。结果　自制的GIFA纸条法检测 64 5份血清标本 ,与ELISA、RIA法作对比 ,符合率分别为 97.4%、96.9%、10 0 .0 1% ,最低检测浓度分别为 2ng/ml、1ng/ml、1ng/ml。 结论　本实验方法具有较高的特异性和灵敏度 ,操作简便。     

Guns and Knives in New Mexico: Patterns of Penetrating Trauma, 1978–1993

Objective: To identify patterns of nonfatal and fatal penetrating trauma among children and adults in New Mexico using ED and medical examiner data.
Methods: The authors retrospectively sampled in 5-year intervals all victims of penetrating trauma who presented to either the state Level-1 trauma center or the state medical examiner from a 16-year period (1978–1993). Rates of nonfatal and fatal firearm and stabbing injury were compared for children and adults.
Results: Rates of nonfatal injury were similar (firearm, 34.3 per 100,000 person-years; stabbing, 35.1). However, rates of fatal injury were significantly different (firearm, 21.9; stabbing, 2.7; relative risk: 8.2; 95% confidence interval: 5.4, 12.5). From 1978 to 1993, nonfatal injury rates increased for children (p = 0.0043) and adults (p < 0.0001), while fatal penetrating injury remained constant. The increase in nonfatal injury in children resulted from increased firearm injury rates. In adults, both stabbing and firearm nonfatal injury rates increased.
Conclusions: Nonfatal injury data suggest that nonfatal violence has increased; fatal injury data suggest that violent death rates have remained constant. Injury patterns vary by age, mechanism of trauma, and data source. These results suggest that ED and medical examiner data differ and that both are needed to guide injury prevention programs.     

Dissociable correlates of two classes of retrieval processing in prefrontal cortex

Although substantial evidence suggests that the prefrontal cortex (PFC) implements processes that are critical for accurate episodic memory judgments, the specific roles of different PFC subregions remain unclear. Here, we used event-related functional magnetic resonance imaging to distinguish between prefrontal activity related to operations that (1) influence processing of retrieval cues based on current task demands, or (2) are involved in monitoring the outputs of retrieval. Fourteen participants studied auditory words spoken by a male or female speaker and completed memory tests in which the stimuli were unstudied foil words and studied words spoken by either the same speaker at study, or the alternate speaker. On "general" test trials, participants were to determine whether each word was studied, regardless of the voice of the speaker, whereas on "specific" test trials, participants were to additionally distinguish between studied words that were spoken in the same voice or a different voice at study. Thus, on specific test trials, participants were explicitly required to attend to voice information in order to evaluate each test item. Anterior (right BA 10), dorsolateral prefrontal (right BA 46), and inferior frontal (bilateral BA 47/12) regions were more active during specific than during general trials. Activation in anterior and dorsolateral PFC was enhanced during specific test trials even in response to unstudied items, suggesting that activation in these regions was related to the differential processing of retrieval cues in the two tasks. In contrast, differences between specific and general test trials in inferior frontal regions (bilateral BA 47/12) were seen only for studied items, suggesting a role for these regions in post-retrieval monitoring processes. Results from this study are consistent with the idea that different PFC subregions implement distinct, but complementary processes that collectively support accurate episodic memory judgments.     

Interprofessional approaches to creating safe,high quality health care

Three supplementary perspectives are presented arguing that interprofessional collaboration is both necessary and desirable. Nonetheless, there are often too many serious intra-professional barriers and obstacles to interprofessional collaboration to make it successful. Some of these barriers, it is argued and illustrated, are found in the multiple ways in which professional identity is tacitly acquired and embodied in the practitioners' habitual, everyday practice. The paper then explores ways in which reflection, especially Second order reflection, can help to elucidate and overcome these obstacles, as well as increasing professional adaptability and competence.     

Systematic Reviews Published in the January 2011 Issue of The Cochrane Library

ABSTRACT

The Cochrane Library of Systematic Reviews is published quarterly as a DVD and monthly online. The January 2011 issue (first quarterly DVD for 2011) contains 4515 complete reviews, 1985 protocols for reviews in production, and 13,521 one-page summaries of systematic reviews published in the general medical literature. In addition, there are citations of 641,000 randomized controlled trials, and 14,018 cited papers in the Cochrane methodology register. The health technology assessment database contains over 9300 citations. One hundred and seven new reviews have been published in the last 3 months, of which five have potential relevance for practitioners in pain and palliative medicine.     

Modified micro-superior percutaneously-assisted total hip: early experiences & case reports

Because of the extensile nature and familiarity of the standard posterior-lateral approach to the hip, a family of "micro-posterior" approaches has been developed. This family includes the Percutaneously-Assisted Total Hip (PATH) approach, the Supercapsular (SuperCap) approach and a newer hybrid approach, the Supercapsular Percutaneously-Assisted Total Hip (SuperPATH) approach. Such approaches should ideally provide a continuum for the surgeon: from a "micro" (external rotator sparing) posterior approach, to a "mini" (external rotator sacrificing) posterior approach, to a standard posterior approach. This could keep a surgeon within his comfort zone during the learning curve of the procedure, while leaving options for complicated reconstructions for the more practiced micro-posterior surgeons. This paper details one author's experiences utilizing this combined approach, as well as permutations of this entire micro-posterior family of approaches as applied to more complex hip reconstructions.     

Determination of creatine kinase isoenzyme MB activity in serum using immunological inhibition of creatine kinase M subunit activity. Activity kinetics and diagnostic significance in myocardial infarction.

This is a new method for the determination of creatine kinase isoenzyme MB activity in serum. The method uses direct activity measurement of creatine kinase B subunit activity after blocking of CK-M subunit activity by inhibiting antibodies. The test takes no longer than 15 min. The method yields an intra-serial C.V. of 2.0-12.9%, and a C.V. from day to day of 5.5%. The detection limit is 3.4 U/l creatine kinase MB. In the 95 cases with proven myocardial infarction several types of creatine kinase MB activity kinetics could be determined. The percentage of creatine kinase MB of peak CK-total is 6-25%, with a mean of 11.1%. The amount of creatine kinase MB with respect to total CK activity after reinfarction is higher than the amount after initial infarction.     

Structure and function of "metalloantibiotics"

Although most antibiotics do not need metal ions for their biological activities, there are a number of antibiotics that require metal ions to function properly, such as bleomycin (BLM), streptonigrin (SN), and bacitracin. The coordinated metal ions in these antibiotics play an important role in maintaining proper structure and/or function of these antibiotics. Removal of the metal ions from these antibiotics can cause changes in structure and/or function of these antibiotics. Similar to the case of "metalloproteins," these antibiotics are dubbed "metalloantibiotics" which are the title subjects of this review. Metalloantibiotics can interact with several different kinds of biomolecules, including DNA, RNA, proteins, receptors, and lipids, rendering their unique and specific bioactivities. In addition to the microbial-originated metalloantibiotics, many metalloantibiotic derivatives and metal complexes of synthetic ligands also show antibacterial, antiviral, and anti-neoplastic activities which are also briefly discussed to provide a broad sense of the term "metalloantibiotics."