高效液相色谱-荧光检测法快速测定脊髓和脑内氨基酸

目的建立快速测定脊髓和脑内氨基酸浓度的方法。方法通过邻苯二甲醛和β-巯基乙醇柱前衍生、反相高效液相色谱结合荧光(激发波长280nm,发射波长340nm)检测氨基酸浓度。用0.1mol/L磷酸盐缓冲液(pH值6.00~6.25)和甲醇(46%)混合液(加入四氢呋喃2%)作流动相,流速为1ml/min。结果在0.1~40.0μmol/L范围内,氨基酸浓度与相对峰面积之间的相关系数(r)均值为0.9992,日内和日间变异系数(CV)分别为2.41%~5.23%、3.51%~6.48%,平均回收率为90.13%~98.47%。结论本法灵敏度高、快速、简便易行,可用于脊髓和脑内氨基酸测定。     

毛细管电泳快速测定脑脊液中谷氨酰胺含量

目的：建立毛细管区带电泳快速测定脑脊液中谷氨酰胺的方法。方法：在碱性条件下，谷氨酰胺与2，4－二硝基氟苯（DNFB）反应生成DNFB氨基酸衍生物，以pH9.8的缓冲液电泳分离，214nm波长测定衍生产物。结果：衍生产物可在5min内分离测定，在101－2025μmol/L范围内呈线性，r=0.9975,批内CV3．9％，批间CV6．1％，平均回收率97．18％，最低检测浓度11μmol/L。结论：毛细管区带电泳法具有快速简便、准确、灵敏的特点，是临床及科研测定谷氨酰胺的理想方法。     

高效毛细管电泳法快速测定血清中游离氨基酸

目的 建立快速测定人血清中游离氨基酸的方法.方法 用高效毛细管电泳法对40例健康体检者及32例肾病患者血清氨基酸进行检测.结果 8 min内分离16种氨基酸,线性范围在10～700 μmol/L,最低检出限为2.5～7.9 μmol/L,回收率86.3%～107.4%,批内精密度2.8%～10.3%,批间精密度3.5%～11.6%.对32例肾脏病患者血清中氨基酸测定结果与40例健康对照组氨基酸测定结果进行统计学分析.结论 高效毛细管电泳法具有快速、准确、灵敏等特点,为临床及科研工作中测定氨基酸提供了一种有效的方法.     

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。     

应用高效毛细管电泳技术快速分离尿肌酐

目的建立1种快速测定尿中肌酐浓度的毛细管电泳方法.方法用pH2.5、0.1 mmol/L磷酸作为电泳缓冲液,利用非涂层石英毛细管48.5 cm×50 μm(id),采用外标法定量,检测波长191 nm.结果本法在34.5～8 840 μmol/L浓度范围内呈良好的线性(r=0.998),日内和日间变异系数分别为3.5%、6.2%,平均回收率94.7%.与苦味酸法有良好的相关性(Y=1.004X+0.004,r=0.998,n=45).结论本法线性范围宽、简单、快速,可应用于临床样品的检测.     

血清钙的偶氮氯膦Ⅰ测定法

目的建立一种简便、灵敏、试剂稳定的分光光度法测定血清钙.方法用国产试剂偶氮氯膦Ⅰ作显色剂测定血清钙.结果显色络合物最大吸收波长为580 nm,线性范围达4.5 mmol/L,摩尔吸光系数为7.72×103L·mol-1·cm-1.回收率为98.3%～101.3%,批内变异系数和批间变异系数分别为1.87%与2.13%,与原子吸收分光光度法y比较具有良好的相关性,Y=1.075X-0.120,r=0.984.Mg+3 mmol/L、Cu2+、Fe3+、Zn2+各100μmol/L、胆红素280μmol/L甘油三酯4.8 mmol/L、血红蛋白2.5 g/L以下对本法测定结果无影响.结论该方法具有快速、简便、灵敏可靠等优点,适合于临床应用.     

血清锌自动化分析法的临床应用

1　材料和方法1.1　仪器　日立 7170 A全自动生化分析仪 (日本产 )。1.2　试剂　 1锌标准液 :30 .6μmol/ L ;2试剂 :购买中美合资宁波亚太生物技术公司生产的试剂盒。1.3　方法　日立 7170 A测定参数 :标准液浓度 30 .6 ,标本量5 .0μL ,试剂量 10 0μl测定方法 1点法 ,副 /主波长 70 0 nm /5 70 nm,测定点 (10 ) (5 )。2　结果2 .1　波长选择　锌与络合剂 5 - Br PAPS反应产生紫红色络合物的最大吸收波长为 5 6 0 nm,本仪器无此波长 ,故选用 5 70 nm为主波长。2 .2　标准曲线　按本方法测定 ,锌含量在 2～ 6 0μmol/ L范围内观察仪器…     

HPLC法测定奈韦拉平的血药浓度

目的:建立高效液相色谱法测定奈韦拉平血药浓度的方法。方法:采用Hypersil C18柱(150 mm×4.6 mm,0.5μm);流动相∶乙腈:0.01 mol/L磷酸二氢钠(含0.01 mol/L三乙胺,磷酸调pH至5.0)=18∶82;流量:1.0mL/min;检测波长:240nm。结果:线性范围0.1~5.0 mg/L(r=0.9996);最低检测浓度为0.1 mg/L;日内、日间相对标准差(RSD%)均小于10%;低、中、高三个浓度的提取回收率分别为99.98%、99.41%和100.53%。结论:该法简便、快速、准确,专属性高,适用于奈韦拉平药动学和生物等效性研究。     

高效液相色谱法测定人血清甜菜碱方法的建立

目的 采用柱前衍生高效液相色谱法-紫外检测法建立一种测定人血清甜菜碱浓度的方法.方法 以对溴苯乙酰溴和18-冠醚-6溶于乙腈制成衍生液,将其与甜菜碱反应形成的衍生物用Supelcosil LC-SCX色谱柱进行分离.流动相为乙腈∶水体积比90∶10,含16 mmol/L的氯化胆碱,等度洗脱,流速为0.8 ml/min,检测波长为259 nm,利用甜菜碱标准品绘制标准曲线,外标法定量检测20名健康大学生志愿者血清甜菜碱.结果 甜菜碱的测定线性范围为6.25～200.00 μmol/L,回归方程Y=1 568.1X-2 747.5,R2=0.999 8.最低检测限为3.0 μmol/L.批内不精密度为1.88%～3.79%(平均3.24%),批间不精密度为3.14%～6.76%(平均4.39%).方法 回收率为95.89%～102.86%(平均99.16%).结论 成功建立一种检测人血清甜菜碱浓度的方法,适用于实验室及临床常规检测.     

高效液相色谱法测量患儿血清甲氨蝶呤浓度

目的：建立测定患儿血清中甲氨蝶呤浓度的方法,用于临床患儿血药浓度监测。方法色谱柱采用岛津原装柱（250 mm×4．6 mm,5μm）,流动相为0．025 mol/L 磷酸二氢钠溶液（pH 为5．41）-甲醇（76∶24,V/V）,流速1．0 mL/min,检测波长313 nm,柱温35℃,样品经10％高氯酸沉淀蛋白后,取上清液130μL 加10μL 1 mol/L NaOH 后直接进样。结果血浆中甲氨蝶呤的浓度在0．00～4．84μmol/L 和4．84～10．00μmol/L 范围内与峰面积线性关系良好,r 1＝1,r2＝1,平均回收率分别为61．67％和71．83％,日内及日间精密度的 RSD 小于10％和12％（n＝3）。结论所用方法简便快速、准确可靠、灵敏度高、重现性好、线性范围宽和成本低,适于甲氨蝶呤的血药浓度监测。     

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.     

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."     

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.