高效液相色谱法测定血浆肝素含量

为探讨高效液相色谱（ＨＰＬＣ）法直接测定血浆游离肝素含量的可行性及其意义，用ＨＰＬＣ分离血浆普通肝素（ｕｎｆｒａｃｔｉｏｎａｔｅｄｈｅｐａｒｉｎ）钠，肝素钙以及低分子量肝素（ＬＭＷＨ），探讨ＨＰＬＣ法所选色谱方法为Ｃ－１８反相柱分离，流动相为甲醇与醋酸铵，外标法定量，紫外肖吸收波长为２１５ｎｍ，回收率为９０．４％～９６．８％，血浆肝素含量在１００～６５００Ｕ／Ｌ之间呈良好线性关系，最低检测限为３０     

血浆游离肝素的胶束动电毛细管电泳分析

目的探讨胶束动电毛细管电泳法（ＭＥＣＣ）测定血浆中游离肝素含量的可行性及意义。方法利用最新的高效毛细管电泳分离技术，用外标法定量，测定血浆中肝素；并对其回收率、线性范围、最低检测限、批内与批间变异系数进行实验，并与凝血酶进行比较。结果建立胶束动电毛细管电泳测定血浆中游离肝素含量方法，并进行了初步临床应用。该方法的回收率为９５．６％～９８．７％；血浆肝素含量在８０～７０００Ｕ／Ｌ之间呈良好的线性关系，最低检测限为２５Ｕ／Ｌ。批内与批间变异系数分别小于３．１％和４．５％。结论ＭＥＣＣ检测血浆中肝素是一种简单、快速的新方法。此法适宜于肝素含量高的血样测定     

凝血酶法测定血浆肝素含量及其临床应用

为了保证肝素的疗效及降低出血的危险性，用试管法和仪器法研究了凝血酶时间（ＴＴ）与血浆肝素含量之间的关系，发现在一定浓度的凝血酶作用下，血浆肝素含量在相应范围内与凝固时间的对数值呈良好的线性关系（直线回归ｒ＝０．９７～０．９９），批内变异系数０．８％～２．８％，批间变异系数为１．６％～２．４％。其中凝血酶浓度为１００００Ｕ／Ｌ的工作曲线适应性较好，能满足各种剂量的检测。尚建立了同时测定患者原血浆与对倍稀释血浆肝素含量的方法，扩大了检测范围，最高达血浆肝素含量６４００Ｕ／Ｌ，且能用于纤维蛋白原或抗凝血酶－Ⅲ严重减少时的肝素含量检测。该方法适用于多种肝素制剂（肝素钠、肝素钙和低分子量肝素）。使用上述方法对接受肝素治疗的患者进行了血药浓度的观察，发现按公斤体重给药血中肝素含量的差异较大，疗效难以掌握，应实行剂量个体化的给药原则。大剂量给药（如体外循环）需用鱼精蛋白中和试验时，该方法较传统的肝素中和试验（鱼精蛋白法）更为准确。     

凝血酶法测定血浆肝素含量及其临床应用

为了保证肝素的疗效及降低出血的危险性，用试管法和仪器法研究了凝血酶时间（ＴＴ）与血浆肝系含量之间的关系，发现在一定浓度的凝血酶作用下，血浆肝素含量在相应范围内与凝固时间的对数值呈良好的线性关系（直线回归ｒ＝０．９７～０．９９），批内变异系数０．８％～２．８％，批间变异系数为１．６％～２．４％，其中凝血酶浓度为１００００Ｕ／Ｌ的工作曲线适应性较好，能满足各种剂量的检测，尚建立了同时测定患者原血浆与对     

高效液相色谱荧光检测法测定尿雌三醇

报告用高效液用色谱（ＨＰＬＣ）测定孕妇尿内总雌三醇（Ｅ＿３）的方法。尿标本在酸水解后用乙醚提取Ｅ＿２，蒸干乙醚，残留物用流动相（甲醇／乙腈／水＝４０／１７／４３）重组。标本成分Ｃ＿８柱分离，以２８５ｎｍ波长激发，在６１０ｎｍ测定Ｅ＿３的自然荧光。本法批内ＣＶ为１２％～１．５％（ｘ＝３２．７～１２８．２ｕｍｏｌ／Ｌ），批间ＣＶ别为２．３％和４０％（分别是３８．４和９８．９ｕｍｏｌ／Ｌ）。本法在４．３３～１３８．７ｕｍｏｌ／Ｌ（１２５～４０．００ｍｇ／Ｌ）间呈线性，最低检出１．９ｕｍｏｌ／Ｌ（０．５２ｍｇ／Ｌ，信号／噪声比为３）。     

高效液相色谱法测定血浆同型半胱氨酸

目的 建立测定血浆同型半胱氨酸的高效液相色谱（ＨＰＬＣ）方法。方法 血浆同型半胱氨酸经巯基乙醇还原后用碘乙酸封闭巯基,。再与邻苯二甲醛（ＯＰＡ）反应完成衍生化,以磺在同型半胱氨酸为内标,经Ｃ１８反相柱进行色谱分离,荧光检测器测定结果。结果测定线性范围达１５０μｍｏｌ／Ｌ,最低检测限为１．０μｍｏｌ／Ｌ日内不精密度３．７－４．２％,日间不精密度３．８－４．３％;方法回收率９３－１０２％。结论本方法     

脐血血浆中Epo、G-CSF、IL-2、IL-6含量的测定

我们利用放射免疫法（ＲＩＡ）和酶联免疫吸附法（ＥＬＩＳＡ）检测脐血血浆中红细胞生成素（Ｅｐｏ）、粒细胞集落刺激因子（ＧＣＳＦ）、白细胞介素（ＩＬ）２、ＩＬ６等造血细胞生长因子,报道如下。材料和方法１　脐血血浆　取自２０名健康产妇足月顺产脐血,孕期３８～４０周,以（１５～２０）×１０３Ｕ／Ｌ肝素抗凝,常规方法分离脐血血浆。２　正常成人外周血血浆　由２０名健康献血员提供,以（１５～２０）×１０３Ｕ／Ｌ肝素抗凝,常规方法分离成人外周血血浆。３　试剂来源　由深圳晶美生物工程有限公司提供。４　检测方法…     

人凝血因子Ⅸ∶Ag酶联免疫测定法的建立

采用抗人凝血因子Ⅸ单克隆和多克隆两种抗体，建立了人凝血因子Ⅸ抗原（ＦⅨ∶Ａｇ）酶联免疫测定法（ＥＬＩＳＡ）。标准曲线相关系数为０．９９３（Ｐ＝０．００１，ｎ＝５），测定范围（６．２５～１００）Ｕ／Ｌ，最低检测量１．５Ｕ／Ｌ；批内及批间变异系数分别为７．２５％和４．７３％，回收率９９．７％；与人血浆中其它蛋白质未见交叉反应。测得２０人份正常人血浆中ＦⅨ∶Ａｇ为（１０２７±１７２）Ｕ／Ｌ，与ＦⅨ促凝活性（ＦⅨ∶Ｃ）测得值比较，差异无显著性（Ｐ＞０．５）。重症乙型血友病人血浆ＦⅨ∶Ａｇ测得值与临床诊断相符。     

钼酸铵显色法测定血清过氧化氢酶

介绍一种简便的以钼酸铵显色的血清过氧化氢酶分光光度测定法。该法酶活性在１００ｋＵ／Ｌ内呈良好线性。批内变异ＣＶ＝３．６％，天间变异ＣＶ＝４．５７％。平均回收率为１００．１４％。高ＴＧ、高Ｂｉｌ血清标本，可使Ａ值增加，但不影响Ｃａｔ测定值。ＶｉｔＣ、肝素均可使酶活性减低。由于红细胞内富含Ｃａｔ，溶血标本能显著增加酶活性。血清标本酶活性在－４℃４８ｈ内无变化，在－２５℃能保持５～６周。３８８例体检健康者血清Ｃａｔｘ＝５７．６１ＫＵ／Ｌ，经Ｄ检验，当Ｐ＝０．０５时，属正态分布；Ｐ＝０．１时，属非正态分布。中位数（Ｍｄ）＝５６．２２ＫＵ／Ｌ，百分位数法计算，其９５％范围为２１．５８～１０８．８７（ｋＵ／Ｌ）。初步观察１１９例患者Ｃａｔ活性，显示各类癌症患者Ｃａｔ活性明显减低。     

3－磷酸甘油脱氢酶偶联法测定血清醛缩酶活性

探讨用３－磷酸甘油脱氢酶偶联法测定血清中醛缩酶（ＡＬＤ）活性的最佳反应条件。反应体系的终末浓度：ＴＥＡ１００ｍｍｏｌ／Ｌ，ＦＤＰ４ｍｍｏｌ／Ｌ，碘乙酸０．２２ｍｍｏｌ／Ｌ，ＮＡＤＨ０．２６ｍｍｏｌ／Ｌ，ＧＤＨ≥１０００Ｕ／Ｌ，ＴＰＩ≥１５００Ｕ／Ｌ，ＬＤ≥１０００Ｕ／Ｌ。最适ｐＨ在７．８～８．０，Ｋｍ为７．２×１０－３ｍｍｏｌ／Ｌ。批内ＣＶ：酶活性在７．３４Ｕ／Ｌ和６５．０６Ｕ／Ｌ时，ＣＶ分别为５．７％和１．４％；批间ＣＶ：酶活性在１１．８９Ｕ／Ｌ和１００．０８Ｕ／Ｌ时，ＣＶ分别为６．０％和３．３％，酶活性线性范围至少可达１８０Ｕ／Ｌ。健康人６０名，ＡＬＤ活性为４．５３±１．１７（ｘ±ｓ）Ｕ／Ｌ，男女两组均值无显著性差异。本文对ＴＥＡ－ＨＣｌ（ｐＨ８．０）、Ｔｒｉｓ－ＨＣｌ（ｐＨ８．０）和Ｃｏｌｉｄｉｎｅ－ＨＣｌ（ｐＨ７．５）三种缓冲液用于ＡＬＤ活性测定效果进行了评价，结果表明在ＴＥＡ缓冲液中所测ＡＬＤ活性最高；ＴＥＡ和Ｃｏｌｉｄｉｎｅ两种缓冲液的浓度在２５～１５０ｍｍｏｌ／Ｌ范围内对ＡＬＤ活性无影响，Ｔｒｉｓ缓冲液在５０ｍｍｏｌ／Ｌ时测得酶活性较高，缓冲液浓度过高或过低，酶活性均有所下降     

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.     

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.     

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.     

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.     

俯卧位通气对高海拔地区肺复张治疗无效急性呼吸窘迫综合征患者氧合的影响

目的 探讨俯卧位通气对高海拔地区肺复张术(RM)治疗无效急性呼吸窘迫综合征(ARDS)患者的治疗作用.方法 从海拔2260m的地区医院筛选RM治疗无效的41例ARDS患者[平均氧合指数( PaO2/FiO2)较RM前升高＜20％视为RM无效],依不同病因分为肺内源性ARDS组(ARDSp组)和肺外源性ARDS组(ARDSexp组),每组再按信封法随机分为俯卧位组和仰卧位组,即ARDSp俯卧位组(11例)、ARDSp仰卧位组(9例)、ARDSexp俯卧位组(10例)、ARDSexp仰卧位组(11例).在通气前及通气1、2、3、4h监测动脉血氧分压( PaO2)、PaO2/FiO2、静态顺应性(Cst)、气道阻力(Raw)的变化.结果 通气lh时,ARDSexp俯卧位组PaO2/FiO2( mm Hg,l mm Hg=0.133 kPa)即较通气前显著升高(157.4±40.6比129.3±48.7,P＜0.05),并随通气时间延长呈持续增高趋势,4h达峰值(219.1 ±41.1);且ARDSexp俯卧位组通气3h内PaO2/FiO2较其他3组显著增高,另3组间则差异无统计学意义.ARDSp俯卧位组、ARDSexp俯卧位组通气4h时PaO2/FiO2均较相应仰卧位组显著增高(208.8±39.7比127.4±47.1,219.1±41.1比124.9±50.8,均P＜0.05).4组通气前后Cst无显著改变,各组间差异也无统计学意义.ARDSp俯卧位组通气4h时Raw(cmH2O·L-1·s-1)较通气前显著降低(6.8±1.7比10.7±1.8,P＜0.05),且明显低于其他3组;其他3组各时间点Raw组内及组间比较差异均无统计学意义.结论 俯卧位通气作为ARDS机械通气重要策略之一,可以改善RM无效高原ARDS患者的氧合,为抢救患者赢得宝贵的时间.     

Digital imaging among medical facilities

The Department of Veterans Affairs (VA) in the USA operates a network of 172 medical centres which all utilize a hospital information system (HIS) which has been developed and is currently maintained by the VA. During the past several years, an image management and communication module has been developed, installed and clinically utilized at the Washington DC and Maryland VA Medical Centres. This image management and communication system, referred to as the decentralized hospital computer program (DHCP) imaging system, is fully integrated with a commercial picture archiving and communication system (PACS). The system is utilized to capture, archive, and display all images generated within the hospital including radiology, nuclear medicine, pathology, endoscopy, bronchoscopy, and dermatology, intraoperative photographs, ECG data, and a limited number of paper documents. The ultimate goal of the project is to have all patient text and image data available at any clinical workstation to any authorized user anywhere within the network of medical centres. Clinical requirements for an imaging workstation include ease of use, rapid and reliable access to the complete set of patient information, and images which are of acceptable quality to meet the requirements of the user and the subspecialty. Patient confidentiality and data security must be safeguarded at all times. Integration of the images with the remainder of the patient's database was found to be critical to the success of the project. The experience at the Washington and Maryland facilities suggests that an imaging system that is successfully integrated with a hospital information system can provide substantial clinical and economic benefits both within and among medical centres. Clinical acceptance and utilization of the system has been excellent, particularly in diagnostic radiology where DHCP Imaging has been interfaced to a commercial PAC system. Based upon this initial experience, the VA has begun to deploy the system throughout its large network of medical centres.     

Myocardial elastography at both high temporal and spatial resolution for the detection of infarcts

Myocardial elastography is a novel method for noninvasively assessing regional myocardial function, with the advantages of high spatial and temporal resolution and high signal-to-noise ratio (SNR). In this paper, in-vivo experiments were performed in anesthetized normal and infarcted mice (one day after left anterior descending coronary artery [LAD] ligation) using a high-resolution (30 MHz) ultrasound system (Vevo 770, VisualSonics Inc., Toronto, ON, Canada). Radiofrequency (RF) signals of the left ventricle (LV) in longitudinal (long-axis) view and the associated electrocardiogram (ECG) were simultaneously acquired. Using a retrospective ECG gating technique, 2-D full field-of-view RF frames were acquired at an extremely high frame rate (8 kHz) that resulted in high-quality incremental displacement and strain estimation of the myocardium. The incremental results were further accumulated to obtain the cumulative displacements and strains. Two-dimensional and M-mode displacement images and strain images (elastograms), as well as displacement and strain profiles as a function of time, were compared between normal and infarcted mice. Incremental results clearly depicted cardiac events including LV contraction, LV relaxation and isovolumetric phases in both normal and infarcted mice, and also evidently indicated reduced motion and deformation in the infarcted myocardium. The elastograms indicated that the infarcted regions underwent thinning during systole rather than thickening, as in the normal case. The cumulative elastograms were found to have higher elastographic SNR (SNR(e)) than the incremental elastograms (e.g., 10.6 vs. 4.7 in a normal myocardium, and 6.0 vs. 2.4 in an infarcted myocardium). Finally, preliminary statistical results from nine normal (m = 9) and seven infarcted (n = 7) mice indicated the capability of the cumulative strain in differentiating infracted from normal myocardia. In conclusion, myocardial elastography could provide regional strain information at simultaneously high temporal (>/=0.125 ms) and spatial ( approximately 55 microm) resolution as well as high precision ( approximately 0.05 microm displacement). This technique was thus capable of accurately characterizing normal myocardial function throughout an entire cardiac cycle, at the same high resolution, and detecting and localizing myocardial infarction in vivo.