HPLC法测定末梢血中苯丙氨酸与酪氨酸及在苯丙酮尿症中的应用

目的 建立一种简单、快速的反相高效液相色谱法测定末梢血液中的苯丙氨酸(Phe)与酪氨酸(Tyr)并用于苯丙酮尿症(PKU)的筛查.方法 末梢血标本与等量5%(v/v)高氯酸溶液混合处理去除蛋白后离心取上清液20 μl直接进样分析.用Hypersil C8色谱柱(6.0mm×300mm,10μm)分离,流动相为乙腈-水(体积比5:95),流速1.5 ml/min,紫外检测波长210 nm,室温下测定.结果 末梢血中Tyr和Phe均分离良好,保留时间分别为5.88 min和8.43 min.102例健康儿童指血Phe浓度67.7±15.4/μmol/L,Tyr浓度为62.2±13.9μmol/L,Phe/Tyr比值为1.15±0.27.32例正常新生儿足跟血Phe浓度为66.2±20.5 μmol/L,Tyr浓度为59.5±18.8/μmol/L,Phe/Tyr比值为1.12±0.24.结论 该法灵敏度高,特异性好,方法 简便快速,成本低康,适合于末梢血中Phe和Tyr的同时测定及PKU的筛查与监控.     

苯丙酮尿症患儿家庭照顾网上调查分析

苯丙酮尿症(PKU)是常染色体隐性遗传病,由于编码苯丙氨酸羟化酶(PAH)的基因发生突变[1,2],导致肝内苯丙氨酸羟化酶活性缺乏或显著降低,苯丙氨酸(Phe)转化为酪氨酸(Tyr)障碍,血中Phe水平显著升高、Tyr降低,进而影响脑的发育[3].     

高效液相色谱法及串联质谱法在苯丙酮尿症筛查诊断中的应用

目的考查了高效液相色谱(HPLC)法和串联质谱(MS/MS)法在遗传代谢性疾病苯丙酮尿症(PKU)筛查中的应用及意义。方法利用MS/MS法和HPLC法分别分析了1 860例出生3d至11岁儿童的干血滤纸片及全血标本中苯丙氨酸(Phe)、酪氨酸(Tyr)浓度及其比值。结果 MS/MS法和HPLC法Phe线性范围为26.02~101.11μmol/L和32.04~132.50μmol/L,Tyr线性范围为41.50~253.07μmol/L和32.85~111.50μmol/L,平均回收率Phe为97.36%和98.43%,Tyr为96.71%和98.99%,批内CV Phe为4.31%和3.97%,Tyr为4.09%和4.01%,批间CV Phe为5.73%和4.58%,Tyr为6.01%和5.24%。结论两种方法均能灵敏,特异的测定血中Phe和Tyr浓度,满足对PKU筛查及诊断的需要。     

苯丙酮尿症患者血清S-100B蛋白测定的临床意义

目的探讨苯丙酮尿症患者血清中S—100B蛋白的水平及与苯丙氨酸的关系。方法用ELISA方法测定27例初诊苯丙酮尿症患儿、5例复诊患儿及42例正常儿童血清内的S-100B蛋白水平，用高效液相色谱荧光法测定血清内的苯丙氨酸浓度并对两者进行分析比较。结果苯丙酮尿症初诊患者血清S-100B水平（0．92±0．15μg／L）显著高于复诊者（0．32±O．11μg／L）与健康对照组（0．22±0．09μ g／L），差异有统计学显著性意义（P〈0．01）；复诊者高于健康对照组，差异有统计学意义（P〈0．05）。初诊患者血清S-100B的浓度与患者体内苯丙氨酸浓度呈正相关（相关系数0．62，P〈0．01）。结论血清S-100B蛋白的水平可作为苯丙酮尿症患者监测脑损伤的指标。     

细菌抑制法测定血苯丙氨酸筛查新生儿苯丙酮尿症

细菌抑制法测定血苯丙氨酸筛查新生儿苯丙酮尿症浙江海盐县人民医院（３１４３００）郑俊夫，沈云芳海盐县妇保所陈萍苯丙酮尿症是由于苯丙氨代谢障碍引起的遗传性疾病。由于苯丙氨酸羟化酶的先天缺陷，苯丙氨酸不能转化为酪氨酸，尿中出现苯丙酮酸等代谢产物。对新生儿的...     

遗传性疾病患儿筛查中股静脉采血操作护理体会

遗传性甲状腺功能低下症是因先天酶缺陷使甲状腺素合成不足所致,属于单基因遗传病.苯丙酮尿症（Phenylketonu-ria,PKU）是由于苯丙氨酸羟化酶缺乏所致的一种代谢病,属于常染色体隐性遗传病.上述两种疾病的早期确诊对于患儿预后及后期智能发育至关重要,而要获得早期诊断,遗传性甲状腺功能低下症通常需要采用放射免疫法检测血清TSH和T4浓度,而苯丙酮尿症则通常需要对经Cuthrie细菌抑制法筛查阳性者做苯丙氨酸或酪氨酸测定[1].完成以上检测,均需采集静脉血约3mL.     

利用质谱仪测苯丙氨酸和酪氨酸的比值鉴别诊断新生儿高苯丙氨酸血症的灵敏度

为探索电射流串联质谱法测氨基酸用于鉴别诊断不同类型的高苯丙氨酸血症(HPA)的敏感性,对78例苯丙氨酸浓度大于150umol/L新生儿的干血标本进行研究,用电射流串联质谱仪(ESI-TMS)定量分析苯丙氨酸(Phe)和酪氨酸(Tyr),并借助于两者的比值区分苯丙酮尿症(PKUn=23)和无PKU的HPA病人(nonpku-HDAn=35;P＜0.001),灵敏度达100%.一过性HPA新生儿(n=9)的苯丙氨酸和酪氨酸比值,低于nonpku-HPA(P＜0.001).以ESI-TMS测氨基酸,提供一个鉴别诊断HPA的有效诊断工具,也是用干血斑进行新生儿筛选的适宜方法.     

高效液相色谱-荧光法同时测定血清中的色氨酸和酪氨酸

目的建立一种高效液相色谱（HPLC）-荧光法（FD）同时测定血清色氨酸（Trp）和酪氨酸（Tyr）的方法。方法色谱条件：Megres C18色谱柱（250 mm×4.6 mm,内径为5μm）,流动相为10%乙腈溶液,流速为1.2 mL/min,Tyr和Trp的荧光检测激发波长和发射波长分别为λexTyr=228 nm、λexTrp=285 nm和λemTyr=306 nm、λemTrp=353 nm。血清样本经5%高氯酸溶液去除蛋白质后取上层清液直接进样进行分析测定。且对样本的保存方法进行了探讨。结果 Tyr的保留时间为3.4 min,线性范围为0.275~275μmol/L,最低检出浓度为0.004μmol/L,回收率为90.5%~108.8%。Trp的保留时间为7.6 min,线性范围为0.490~196μmol/L,最低检测浓度为0.005μmol/L,回收率为88.8%~97.2%。Tyr和Trp的日内、日间测定的相对标准偏差均〈5%,苯丙氨酸、5-羟色胺、犬尿喹啉酸、犬尿氨酸和肌酐等物质对该法均无干扰。样本应-20℃冰冻保存。结论该方法简便、快速、敏感、特异,可同时测定血清Tyr和Trp,适合于临床和科研应用。     

PHE/TYR比值在新生儿苯丙酮尿症筛查中的应用

目的 分析Phe/Tyr比值在新生儿苯丙酮尿症筛查中的作用.方法 采用API-300 SPOTCHECK连续流动荧光分析系统检测1103例正常新生儿的筛查血片和27例已经确诊的PKU阳性患儿血片的Phe和Tyr值,并进行分析.结果 API-300 SPOTCHECK连续流动荧光分析法的阳性复合率为103.7%,阴性复合率为99.9%.结论 Phe/Tyr 比值是提高PKU筛查准确度的重要参数.     

苯丙酮尿症患儿家庭照顾网上调查分析

苯丙酮尿症（PKU）是常染色体隐性遗传病，由于编码苯丙氨酸羟化酶（PAH）的基因发生突变^[1,2]，导致肝内苯丙氨酸羟化酶活性缺乏或显著降低，苯丙氨酸（Phe）转化为酪氨酸（Tyr）障碍，血中Phe水平显著升高、Tyr降低，进而影响脑的发育^[3]。在新生儿和婴儿早期无典型的临床表现，当发现症状时已出现精神运动发育迟缓和不可逆的中枢神经系统损害，造成严重的精神障碍^[4]。据顾学范等^[4]的调查报告，我国PKU发病率为1／11144。我国每年新增数千名患儿，PKU给社会和家庭带来沉重的精神和经济负担。目前唯一的治疗方案即为饮食治疗，而饮食治疗主要由家庭中患儿的照顾者来承担。由于此病是一种罕见的遗传性疾病，照顾者缺乏治疗的相关知识，因此对于PKU患儿家庭治疗的关注成为医护人员责无旁贷的使命。然而，对于PKU患儿的家庭照顾还没有走入我们护理人员的视野。本次调查将描述PKU患儿家庭照顾中的一些现况，旨在呼吁护理界人士高度关注这个特殊的人群。现报道如下。     

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.