凝胶抗球蛋白试验的特点及实验分析

目的 分析评价凝胶抗球蛋白试验(GAT)的特点，增加输血安全性。方法 以不同稀释度的商品化抗D(IgG)为基础制备不完全抗体和致敏红细咆为模拟标本，通过一系列实验(并结合文献)分析GAT(DiaMed产品)的特点。结果红细胞洗涤与否不影响GAT检测的结果；外加AB型血浆至40μl后，才使GAT(1：64抗D血清25pl与0．8％O型混合红细胞悬液50μl反应)产生的强阳性略有减弱；与专用稀释液ID-diluent2比较，用生理盐水替代稀释液导致结果阳性程度降低，而换用某厂家的LIM稀释液则出现不同程度的红细胞溶血现象；GAT可测的抗D(IgG)最高效价为1：256，反应灵敏度明显高于试管法，而近似于聚凝胺法。结论 GAT反应过程中无洗涤步骤，使用专用稀释液，检测灵敏度较高。GAT检测中使用配套试剂材料和严格按照说明书操作非常重要。     

IgG抗-A引起新生儿溶血病1例

目的对1例母亲为B型Rh(D)阳性、患儿为AB型Rh(D)阳性,由母体产生IgG抗-A引起的新生儿溶血病进行分析探讨。方法检测母亲及患儿的ABO、Rh(D)血型及不规则抗体筛查、新生儿溶血病三项试验、患儿母亲血清与患儿红细胞的反应性、患儿母亲的抗体效价。结果患儿血型为AB型Rh(D)阳性,患儿母亲为B型Rh(D)阳性,患儿直接抗人球蛋白试验阳性,母亲血清与患儿红细胞反应为阳性,新生儿体内检测到游离IgG抗-A,红细胞放散液中检测到IgG抗-A,母亲IgG抗-A效价为64。结论 B型母亲体内IgG抗-A导致AB型患儿发生ABO血型系统新生儿溶血病。     

流式细胞术检测不同RhD血清型红细胞RhD抗原的表达

本研究旨在建立流式细胞术(FCM)检测红细胞表面RhD抗原的实验方法,并研究不同RhD血清型红细胞表面D抗原表达的差异。选取标准的RhD( )和RhD(-)细胞按1∶1比例混合,用间接标记法[一抗为IgG抗-D,二抗为FITC-抗-IgG F(ab′)2]染色,应用FCM检测RhD( )细胞的比例,并确立IgG抗-D的最佳使用稀释度。采用FCM检测RhD阳性、弱D型、RhDel型、RhD阴性红细胞表面D抗原数量。结果显示,IgG抗-D单克隆抗体的最佳使用稀释度是1∶4,1×106/50μl。RhD阳性、弱D型、RhDel型、RhD阴性者RhD( )红细胞百分率分别为(96.8±2.97)%、(79.5±9.88)%、(47.8±11.43)%、(3.7±2.96)%,红细胞表面D抗原平均荧光强度依次为33.3±6.21道、18.6±5.39道、7.10±1.17道、0.79±0.55道。结论:4种不同血清表现型的RhD抗原数量的差异有显著性,以RhD阳性的抗原数量最多,弱D型次之,RhDel型最少。     

流式细胞术产前非侵入性定量检测胎-母出血的研究

目的　建立产前非侵入性定量检测RhD阴性孕妇胎 母出血的流式细胞术实验方法。方法　RhD阴性红细胞与RhD阳性红细胞按一定比例混合 ,用抗 D作为一抗结合D+ 红细胞 ,以羊抗人IgGF(ab′) 2 FITC作为二抗 ,连接抗 D ,应用流式细胞术检测D+ 红细胞占D-红细胞的比例 ,建立抗 D及抗人IgGF(ab′) 2 FITC的量效关系 ,确定两者最佳使用剂量。在此实验条件下 ,进行流式细胞术检测值与实际值的相关性分析 ,判定可以检测出的D-红细胞中混有D+ 红细胞的准确出血量范围 ,应用建立的流式细胞术方法测定 9例RhD阴性孕妇外周血中的胎 母出血量。结果　抗 D的有效使用剂量为 1∶4 0 0稀释 ,10 0 μl/5× 10 6细胞 ;羊抗人IgGF(ab′) 2 FITC的有效使用剂量为 2 5 μg/5× 10 6细胞 ;流式细胞术测定比例与已知实际比例的相关系数r =0 998;准确检测的胎 母出血量为 (0 .6～ 30 )ml。 9例RhD阴性孕妇外周血中除 2例未检出D+ 细胞而无法判断外 ,其余 7例均检测出胎 母出血 ,胎 母出血量为 (1.2～ 6 .4 8)ml。结论　针对孕期发生胎 母出血反应的RhD阴性孕妇 ,可以应用流式细胞术方法在产前准确无损伤地定量检测胎 母出血量 ,指导临床预防、治疗Rh新生儿溶血病     

免疫性抗D和抗B致新生儿Rh伴ABO溶血1例实验分析

目的 分析非O型母亲相关的免疫性抗D和抗B致新生儿Rh伴ABO溶血的实验室特点,为临床明确诊断提供帮助.方法 对患儿和母亲进行常规实验室检查、改良直接抗人球蛋白试验(改良Coombs试验)、血型检测以及吸收放散试验,红细胞抗体释放试验检测导致患儿红细胞受累的IgG抗体,血清游离抗体试验检测患儿和母亲血清中IgG抗体.结果 患儿血红蛋白140g/L,网织红细胞计数7.8%.总胆红索722.0μmol/L.患儿血型为B型CcDee,红细胞抗体释放试验抗D阳性和抗B弱阳性.血清游离抗体试验抗D阳性.母亲为第四胎第二产,血型为A型ccdee,血清游离抗体试验抗D阳性.结论 患儿红细胞上存在的大量IgG抗体型抗D和部分抗B是导致发生新生儿溶血病(HDN)的主要原因.A型母亲由于多次妊娠使体内累积IgG型抗B抗体并导致患儿发生HDN,提示临床对于有多次妊娠史的非O型孕妇.需考虑其婴儿发生HDN的可能性.     

输血相容性检测室内质控品制备技术优化与性能评价

目的优化自制输血相容性检测室内全血质控品制备技术,检测其保存过程中综合性能指标的变化,确定合理的处理流程、保存期限并评价其应用价值。方法选择多人份采集时间≤10 d的B型RhD阴性健康献血者标本,将其混合后离心留取上清血浆,再将混合浓缩红细胞分成2组,MAP组:使用MAP红细胞保养液洗涤2次;Saline组:使用生理盐水洗涤2次。将2组洗涤后的红细胞分别与MAP红细胞保存液、对应混合血浆按照1∶2∶3的体积比混合。选择商品化IgG抗-D试剂做抗-D效价测定,确定出现最后1个2+凝集强度的稀释倍数,并按照此稀释倍数分别在2组质控品中填加相应体积的IgG抗-D。将2组混合悬液分装在硬质塑料试管中盖帽4℃保存,每天在室温放置1 h,分别在保存的0、35、42、49 d检测质控品标本红细胞与标准抗-B的凝集强度、IgM抗-A与反定A细胞的凝集强度、IgG抗-D与RhD阳性O型红细胞的凝集强度、上清液中Na+、K+、LDH、乳酸、FHb浓度、红细胞形态变化以及质控品中细菌繁殖情况。结果保存过程中2组质控品红细胞B抗原、IgG抗D抗体反应活性均无明显变化(P>0.05),IgM抗A抗体反应活性虽出现波动(P<0.01),但凝集强度变化均在1+范围内;2组质控品K+、乳酸浓度均随保存时间延长明显增高(P<0.01),但保存35、42d时2组各指标之间无明显差异(P>0.05);2组质控品FHb浓度均随保存时间延长而增高,但相同保存时间2组之间比较无明显差异(P<0.01)。保存49 d时MAP组和Saline组FHb浓度分别为(857.1±301.5)mg/L和(595.3±334.9)mg/L,与保存0d相比均明显升高(P<0.01),MAP组部分质控品FHb浓度已经超过中度溶血标准(1 000 mg/L);2组质控品保存末期(≤42 d)红细胞都会出现一定程度的皱缩并形成棘突,但2组之间无明显差异;2组质控品保存过程中都未见细菌生长。结论本室采用2种方法自制的全血质控品质量无明显差异,保存期都能达到42 d,且管间差异小、抗原抗体反应活性稳定,能够满足输血相容性检测室内质控的相关要求,适合在输血相容性检测实验室推广。     

联合进口与国产试剂检测献血者抗-HCV的必要性分析

目的　探讨5种不同厂家酶免疫试剂的灵敏度和联合应用的必要性。方法　将2 0份抗 HCV阳性标本按1∶10 0～1∶15 0 0不同倍数进行稀释,用5种进口与国产酶免疫试剂检测不同稀释倍数抗 HCV阳性标本。结果　2 0份阳性标本经5种不同厂家试剂检测,结果均为阳性,无一漏检。随着2 0份血样稀释倍数增加,阳性标本检出数和检出阳性标本的试剂数逐渐减少,当稀释倍数达到1∶110 0～1∶130 0时,国产与进口试剂在灵敏度上的差异有显著性(P <0 .0 1)。结论　献血者初复检宜分别选用灵敏度不同的国产和进口试剂联合检测,以确保检测结果的可靠性。     

全自动微柱玻璃珠技术提高血型相容性检测效能

探讨基于微柱玻瑞珠技术的ORTHO AutoVue Innova全自动血型及配血分析系统能否满足输血科血型血清学实验要求.用ORTHO AutoVue Innova全自动血型及配血分析系统和盐水介质法分别检测IgM类抗C、抗c、抗D、抗E和抗e系列稀释液各16份与相应抗原阳性的红细胞凝集反应的强度;用ORTHO AutoVue Innova全自动血型及配血分析系统、凝聚胺法和抗人球蛋白法分别检测IgG类抗D稀释液各16份与RhD阳性红细胞凝集反应的强度;再比较分析微柱玻璃珠法与传统对照实验的检测灵敏度.结果表明,ORTHO AutoVue Innova全自动血型及配血法对IgM类抗C、抗c、抗D、抗E和抗e的检测灵敏度分别为1:69.8、1:33.4、1:1448.1、1:139.6和1:32.0;盐水介质法的相应灵敏度分别为1:16.7、1:16.6、1:430.5、1:34.9和1:9.9.2种方法检测灵敏度的差异均具有统计学意义(IgM类抗C、杭c、抗D、抗E及抗e的t值分别为14.38、5.48、10.25、12.65和9.59,p均＜0.05).对于IgG类抗D微柱玻璃珠法、凝聚胺法和抗人球蛋白法的检测灵敏度分别为1:980.6、1:181.0和1:304.4,三种检测方法间灵敏度的差异具有统计学意义(F=51.15,p<0.01).结论:ORTHO AutoVue Innova全自动血型及配血法比传统方法具有更高的检测灵敏度,将其用于常规血型血清学检测是安全可靠的.     

免疫性抗D和抗B致新生儿Rh伴ABO溶血1例实验分析

目的分析非O型母亲相关的免疫性抗D和抗B致新生儿Rh伴ABO溶血的实验室特点．为临床明确诊断提供帮助。方法对患儿和母亲进行常规实验室检查、改良直接抗人球蛋白试验（改良Coombs试验）、血型检测以及吸收放散试验。红细胞抗体释放试验检测导致患儿红细胞受累的IgG抗体,血清游离抗体试验检测患儿和母亲血清中IgG抗体。结果患儿血红蛋白140g/L,网织红细胞计数7．8％,总胆红索722．0μmol／L。患儿血型为B型CcDee,红细胞抗体释放试验抗D阳性和抗B弱阳性,血清游离抗体试验抗D阳性。母亲为第四胎第二产,血型为A型ccdee,血清游离抗体试验抗D阳性。结论患儿红细胞上存在的大量IgG抗体型抗D和部分抗B是导致发生新生儿溶血病（HDN）的主要原因。A型母亲由于多次妊娠使体内累积IgG型抗B抗体并导致患儿发生HDN,提示临床对于有多次妊娠史的非O型孕妇,需考虑其婴儿发生HDN的可能性。     

流式细胞计数法和试管疑集法检定红细胞相关C3d的初步比较

该研究比较了检定红细胞相关补体和免疫球蛋白的流式细胞计数法和试管凝集法。对20例病人用抗—C3d、抗IgG、抗—IgM和抗—IgA作流式细胞计数分析试管法直接抗人球蛋白试验(DATs)阳性的红细胞。对l例胎儿淋巴细胞溶血危险的病人连续采样进行了测试。抗IgG的流式细胞计数法和试管法的结果如下：20例中的12例样品两种方法都有反应，20例中的6例两种方法都不反应。20例中有两例试管反应与流式细胞计数无反应；抗—C3d的结果显示，20例中有9例两种方法都反应，20例中有11例在试管法有反应和流式细胞计数法的反应不一致；IgM的流式细胞计数中，三份样品与抗—IgM反应。一份来自移植了B型供者的干细胞的A型妇女的样品显示，移植后3—6天里，流式细胞计数检测到与抗—IgG和(或)抗—C3d发生反应，试管法则为阴性反应。结论 流动细胞计数法在检测红细胞相关C3d时灵敏度高于试管法。进一步的研究则需要检测出C3d的水平与临床结果的关系。     

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.     

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.     

Prise en charge des entorses externes de cheville : étude clinique préliminaire sur l’efficacité du dispositif Myolux™

Ankle sprains are the most common injury of the musculoskeletal system and are associated with significant societal and economic impacts. It has been proven that classical therapeutic strategies may not be effective in preventing recurrent injuries: the recurrence rates reported in the literature can reach 73%. In order to provide an effective rehabilitation solution, a destabilizing orthosis was developed. This device is equipped with a mechanical articulator reproducing the subtalar mechanics and placed under the heel. In this paper, we present the main results of a preliminary clinical study conducted between 2004 and 2007. All subjects included in this study were treated with the abovementioned orthosis during 10 rehabilitation sessions of 30 minutes each. Data show a relatively low recurrence rate of 12% for the overall population. Moreover, it's of primary importance to note that this satisfactory ratio is largely reduced (3% of recurrence rate) for the 29 patients who performed one training session per month after the 10th initial rehabilitation sessions. Hence, the destabilizing orthosis appears to be an effective solution to prevent recurrent ankle sprains. However, joint protection requires long-term and regular training sessions. This result has motivated the development of a similar device allowing patients to perform training sessions at home. Finally, data obtained in this study are promising awaiting the final results of the comparative, multicentric and independent clinical trials currently managed by the Hospices Civils de Lyon.     

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.     

Clinical Trial Updates and Hotline Sessions presented at the European Society of Cardiology Congress 2007

This article provides information and commentaries on trials which were presented at the Hotline and Clinical Trial Update Sessions at the European Society of Cardiology Congress 2007 in Vienna. The key presentations were performed by leading experts in the field with relevant positions in the trials or registries. It is important to note that unpublished reports should be considered as preliminary data, as the analysis may change in the final publications. The comprehensive summaries have been generated from the oral presentation and the webcasts of the European Society of Cardiology and should provide the readers with the most comprehensive information of relevant publications.     

Balance and Boundaries in Grief Counseling: An Intervention Framework for Volunteer Training

Volunteers or paraprofessional counselors are commonly used to provide supportive care to the bereaved. These counselors generally are trained in basic listening skills, providing a generic, nonspecific approach to intervention that remains to be proven effective. The present paper outlines a framework that provides paraprofessionals with a broader model for intervention with the bereaved. Attention to boundaries as a helper and balance in the grief recovery are central to the model. Interventions are described that provide the paraprofessional counselor with more options for tailoring their counseling strategy to the individual. These include techniques that are presumed to be more specific to the enhancement of grief recovery.     

A new method for the simultaneous analysis of unmodified and modified urinary nucleosides and nucleobases by high performance liquid chromatography

Details are given of a new, rapid and simple pre-fractionation method and an isocratic high performance liquid chromatography system suitable for parallel analysis of nucleosides and nucleobases from urine and other biological fluids. The quantitative recovery and excellent reproducibility of the method is demonstrated by analysis of representative standard RNA catabolites. The advantage of this new method for application to biological samples is discussed.     

Adsorption of Charged Non-albumin Bound Drugs by Amorphous Silica

We investigated the in vitro drug adsorption of PQ 10150 sodium silicate gel (AIS, Santa Clara, CA) with particle size of 230 um and surface area of 400 nr/g. We observed 99% to 88% adsorption of gentamicin; a mean 91 % of disopyramide; a mean 89% of quinidine at low concentration, falling to 75% at higher concentration. Insulin was 88% adsorbed at low concentrations but less so (65%) at higher concentrations. We observed a mean 83 % adsorption of procainamide, a mean 84% of N-acetyl procainamide, 74% oflidocaine, 73% of amitriptyline; and 44% of desipramine. We found an average 14% reduction of total digoxin concentration when serum containing digoxin (2 to 33 ng/mL) was exposed to sodium silicate, while the reduction in free digoxin concentration was 16%. Five percent ethosuximide was also removed. The adsorption of theophylline, phenobarbital, acetaminophen, phenytoin, ethylene glycol, methotrexate, salicylate, thiocyanate and diazepam was minimal and not significant. We conclude that significant amounts of charged, non-albumin bound drugs can be removed by PQ 10150 sodium silicate gel.