高效液相色谱法快速检测单胺类神经递质及其代谢产物

目的　建立高效液相色谱法快速测定肾上腺素 (E)、去甲肾上腺素 (NE)、多巴胺 (DA)、5′ 羟色胺 (5′ HT)、5 羟吲哚乙酸 (5 HIAA)、高香草酸 (HVA)及多巴柯 (DOPAC) 7种单胺类神经递质及其代谢产物。方法　流动相选用柠檬酸 柠檬酸三钠 甲醇 离子对试剂 EDTA Na2 混合液 (pH 4 .2 ) ;色谱柱采用 30mm× 3mmC18柱 ,用电化学检测器检测。结果　最低检测限为 2～ 30ng/ml,平均回收率为 80 .1%～ 10 7.3%。分别测定 6例正常大鼠及 6例帕金森病 (PD)模型组大鼠的 5种单胺类递质和 12名正常人及 2例嗜铬细胞瘤患者 3种单胺类递质 ,结果皆有明显区别和升高。结论　此法能满足大鼠PD模型的研究和嗜铬细胞瘤患者实验诊断的需要。     

高效液相色谱法快速检测单胺类神经递质及其代谢产物

目的建立高效液相色谱法快速测定肾上腺素(E)、去甲肾上腺素(NE)、多巴胺(DA)、5′-羟色胺(5′-HT)、5-羟吲哚乙酸(5-HIAA)、高香草酸(HVA)及多巴柯(DOPAC)7种单胺类神经递质及其代谢产物.方法流动相选用柠檬酸-柠檬酸三钠-甲醇-离子对试剂-EDTA-Na2混合液(pH 4.2);色谱柱采用30 mm×3 mm C18柱,用电化学检测器检测.结果最低检测限为2～30 ng/ml,平均回收率为80.1%～107.3%.分别测定6例正常大鼠及6例帕金森病(PD)模型组大鼠的5种单胺类递质和12名正常人及2例嗜铬细胞瘤患者3种单胺类递质,结果皆有明显区别和升高.结论此法能满足大鼠PD模型的研究和嗜铬细胞瘤患者实验诊断的需要.     

不同发育时期大鼠脑海马区胆固醇对发育与成熟的意义

背景胆固醇可使体外培养的神经元自身突触数量增加,囊泡数量及递质释放效能亦明显增高.目的建立大鼠脑海马组织中微量胆固醇的高效液相色谱检测方法,观察胆固醇在大鼠脑海马发育和成熟中的作用.设计重复测量实验.材料实验于2004-01/06在武汉大学人民医院神经科学研究室完成.选择健康雄性Wistar乳鼠(出生24 h内)、幼鼠(28～30 d,体质量40～60 g)和成鼠(90～100 d,体质量200～250 g)各10只.方法大鼠在麻醉状态下断头取海马称重,制备匀浆,离心,过滤后采用美国安捷伦公司反相色谱柱(ZORBAX XDB C18 4.6*250 mm 5 μm,Agilent),以乙腈-异丙醇(41)为流动相;流速0.8 mL/min;检测波长208nm.VARIAN Prostar 325紫外检测器和美国VARIAN公司的LCWORKSTATION V6.2色谱数据处理系统检测和处理色谱数据,获得胆固醇色谱图.样品中胆固醇含量(mg/g)=10×0.996 5×样品中胆固醇的峰面积×标准胆固醇的含量(g/L)÷标准胆固醇的峰面积主要观察指标①乳鼠、幼鼠和成鼠脑海马胆固醇含量与其色谱峰面积的关系.②乳鼠、幼鼠和成鼠脑海马胆固醇含量.结果30只大鼠均进入结果分析.①胆固醇含量在0.132 3～2.647 2 g/L时与其色谱峰面积呈良好的线性关系(r=0.999 93).②乳鼠、幼鼠和成鼠脑海马胆固醇含量3组样品的胆固醇含量分别为(2.92±0.03),(11.20±3.41),(12.91±1.25)mg/g,差别均有极显著性意义(P＜0.01).结论用高效液相色谱法检测大鼠脑海马组织中的微量胆固醇灵敏,特异性好,且分析时间短.在大鼠发育过程中,脑海马中胆固醇含量渐增,说明胆固醇对大鼠脑海马结构、功能的发育和成熟可能发挥了关键性的作用.     

慢性运动性疲劳大鼠海马多种单胺类递质水平变化规律的研究

目的探讨慢性运动性疲劳状态下,大鼠海马组织各种单胺类递质的变化特点。方法建立慢性运动性中枢疲劳大鼠模型,造模时间为4周;对照组不采取任何干预措施,与模型组相同条件正常喂养,时间四周。所有实验动物取材部位为大鼠双侧海马,检测方法为高效液相色谱电化学检测法,研究指标选取5-羟色胺(5-HT)、去甲肾上腺素(NE)、肾上腺素(E)和多巴胺(DA)的含量进行定量分析。结果与对照组相比,模型大鼠在慢性运动性中枢疲劳状态下,其海马组织中5-羟色胺含量明显升高(P0.01),同时多巴胺和肾上腺素的含量明显下降,二者均在统计学有显著性差异(P0.01,P0.05);而同样属于单胺类神经递质的去甲肾上腺素的水平则明显升高(P0.01)。结论在中枢性疲劳发生时,大鼠海马内四种重要的单胺类神经递质的水平均发生了改变,与额叶皮质研究发现存在不同,这说明大鼠慢性运动性疲劳状态下中枢神经递质确实发生着明显的改变,同时这种变化在不同脑组织中存在着明显差异。     

慢性运动性中枢疲劳大鼠额叶皮质单胺类递质变化的研究

目的研究慢性运动性疲劳状态下,大鼠中枢神经系统各种单胺类递质的变化规律。方法应用慢性运动性中枢疲劳大鼠模型,取材大鼠额叶皮质,采用高效液相色谱电化学检测的方法,对实验大鼠额叶皮质组织5-羟色胺、肾上腺素、去甲肾上腺素及多巴胺的含量进行定量分析。结果与正常饲养大鼠比较,疲劳大鼠在持续训练条件下发生运动性中枢疲劳后,其额叶皮质5-羟色胺含量明显升高,同时多巴胺的含量明显下降,并在统计学出现非常显著性差异(P0.01);而同样属于单胺类神经递质的肾上腺素和去甲肾上腺素的水平则没有出现统计学差异。结论在中枢性疲劳发生时,中枢神经系统内两种重要神经递质5-羟色胺和多巴胺的水平同时发生了改变,且这两种变化的方向均为抑制性趋势,这也说明该模型大鼠形成了中枢疲劳状态,这种状态的形成引起了脑内递质水平调节机制的变化。     

高效液相色谱法快速检测单胺类神经递质及其代谢产物

背景:单胺类神经递质及其代谢产物对实验研究帕金森病和实验诊断嗜铬细胞瘤具有重要意义,是当今的研究热点。目的:建立高效液相色谱法同时快速测定肾上腺素、去甲肾上腺素、多巴胺、5-羟色胺、5-羟吲哚乙酸、高香草酸及多巴柯7种单胺类神经递质及其代谢产物。设计:病例-对照观察和随机对照动物实验。单位:青岛大学医学院附属医院检验科及青岛大学医学院附属医院海阳分院。对象:实验于2003-10/12在青岛大学医学院生理教研室和青岛大学医学院附属医院检验科完成。①2例嗜铬细胞瘤患者,均经CT,MRI或病理切片和临床证实为该病。12例正常对照均为在校大学生,且经查排除高血压、内分泌系统疾病和其他疾病。②选用健康雌性Wistar大白鼠12只,体质量150～180g。随机数字表法分为2组,正常对照组和帕金森病组,每组大鼠各6只。方法:制备7种物质样品(肾上腺素、去甲肾上腺素、多巴胺、5-羟色胺、5-羟吲哚乙酸、高香草酸及多巴柯)的标准色谱图,同时进行样品精密度和回收率的检测。采用高效液相色谱法检测12例正常对照和2例嗜铬细胞瘤患者24h尿中的儿茶酚胺类物质。同时以6-羟基多巴胺损毁帕金森病组大鼠大脑一侧前脑束,以监测正常和帕金森病大鼠健侧及损毁侧尾壳核区纹状体内单胺类神经递质及其代谢产物的释放特点。主要观察指标:①样品精密度和回收率检测结果。②12例正常对照和2例嗜铬细胞瘤患者24h尿中儿茶酚胺类物质的测定结果。③各组大鼠纹状体内单胺类神经递质及其代谢产物的释放特点。结果:12例正常对照和2例嗜铬细胞瘤患者均获得24h尿中儿茶酚胺类物质检测结果,并进入结果分析。实验动物在实验中无死亡。①高效液相色谱法对肾上腺素、去甲肾上腺素、多巴胺、5-羟色胺、5-羟吲哚乙酸、高香草酸及多巴柯7种单胺类物质的线性范围为2～1000μg/L,最低检测限为2～30μg/L,批内精密度为3.0%～8.6%,批间精密度为3.8%～9.7%。多巴胺的平均回收率为107.3%、去甲肾上腺素为89.4%、肾上腺素为80.1%,其余4种物质为89.2%～95.7%。②2例嗜铬细胞瘤患者24h尿中儿茶酚胺类物质的含量显著高于正常人,其中1例以肾上腺素明显增高为主,另1例以去甲肾上腺素明显增高为主,均高于正常人10倍左右。③帕金森病大鼠健侧尾壳核内多巴胺及其代谢产物多巴柯、高香草酸含量均在正常范围内,损毁侧尾壳核内多巴胺、5-羟色胺、多巴柯及高香草酸均显著降低,差异有显著性意义(P<0.01);5-羟色胺及其代谢产物5-羟吲哚乙酸含量无明显改变。④帕金森病大鼠健侧尾壳核内多巴胺更新率(多巴柯/多巴胺和高香草酸/多巴胺的比值)升高;损毁侧尾壳核内多巴胺更新速度也加快,与正常对照组比较差异均有显著性意义(P<0.05)。结论:高效液相色谱法能满足对大白鼠帕金森病模型的研究和对嗜铬细胞瘤患者实验诊断的需要。     

高效液相色谱法快速测定氨基酸类神经递质

目的建立一种同时快速测定大鼠纹状体中谷氨酸(Glu)、天冬氨酸(Asp)、γ-氨基丁酸(GABA)、甘氨酸(Gly)、牛磺酸(Tau)5种氨基酸类神经递质的高效液相色谱法.方法采用Kromasil C18(4.6×250 mm,5 μm)为色谱柱,丹酰氯为柱前衍生试剂,以曲唑酮为内标,以0.1 mol/L醋酸钠缓冲液∶甲醇＝58∶ 42、14 mmol/L庚烷磺酸钠为流动相等度洗脱.结果在1～200 mg/L范围内,各组分线性关系良好(r=0.999).各氨基酸的平均回收率为79.1%～92.9%.结论本方法简便、快速、准确, 可用于临床大样本测定和研究神经及精神疾病氨基酸类神经递质的改变.     

高效液相色谱法测定生物样品中五羟色胺含量

[目的]建立高效液相色谱法测定生物样品中五羟色胺含量的方法。[方法]采用高效液相色谱一电化学检测法。[结果]HPLC法测定生物样品中五羟色胺的线性范围为0．05～50ng／ml，r=0．9997。平均回收率大于90％，日内和日间相对标准偏差均小于5％。[结论]本法简便、准确、快捷，可用于生物样品中的5-羟色胺的测定。     

高效液相色谱法测定复方五倍子液中没食子酸的含量

[目的]建立复方五倍子液中没食子酸的含量测定方法.[方法]采用高效液相色谱法,WelchromTM C18柱,流动相为甲醇-0.1%磷酸水溶液(15∶85),柱温25℃,流速1.0 mL/min,检测波长为273 nm.[结果]相关系数r=0.9999,平均加样回收率为98.4%,相对标准偏差(RSD)为0.40%,具...     

高效液相色谱法快速检测单胺类神经递质及其代谢产物

背景：单胺类神经递质及其代谢产物对实验研究帕金森病和实验诊断嗜铬细胞瘤具有重要意义,是当今的研究热点.目的：建立高效液相色谱法同时快速测定肾上腺素、去甲肾上腺素、多巴胺、5-羟色胺、5-羟吲哚乙酸、高香草酸及多巴柯7种单胺类神经递质及其代谢产物.设计：病例-对照观察和随机对照动物实验.单位：青岛大学医学院附属医院检验科及青岛大学医学院附属医院海阳分院.对象：实验于2003-10/12在青岛大学医学院生理教研室和青岛大学医学院附属医院检验科完成.①2例嗜铬细胞瘤患者,均经CT,MRI或病理切片和临床证实为该病.12例正常对照均为在校大学生,且经查排除高血压、内分泌系统疾病和其他疾病.②选用健康雌性Wistar大白鼠12只,体质量150～180 g.随机数字表法分为2组,正常对照组和帕金森病组,每组大鼠各6只.方法：制备7种物质样品（肾上腺素、去甲肾上腺素、多巴胺、5-羟色胺、5-羟吲哚乙酸、高香草酸及多巴柯）的标准色谱图,同时进行样品精密度和回收率的检测.采用高效液相色谱法检测12例正常对照和2例嗜铬细胞瘤患者24 h尿中的儿茶酚胺类物质.同时以6-羟基多巴胺损毁帕金森病组大鼠大脑一侧前脑束,以监测正常和帕金森病大鼠健侧及损毁侧尾壳核区纹状体内单胺类神经递质及其代谢产物的释放特点.主要观察指标：①样品精密度和回收率检测结果.②12例正常对照和2例嗜铬细胞瘤患者24 h尿中儿茶酚胺类物质的测定结果.③各组大鼠纹状体内单胺类神经递质及其代谢产物的释放特点.结果：12例正常对照和2例嗜铬细胞瘤患者均获得24 h尿中儿茶酚胺类物质检测结果,并进入结果分析.实验动物在实验中无死亡.①高效液相色谱法对肾上腺素、去甲肾上腺素、多巴胺、5-羟色胺、5-羟吲哚乙酸、高香草酸及多巴柯7种单胺类物质的线性范围为2～1 000μg/L,最低检测限为2～30μg/L,批内精密度为3.0%～8.6%,批间精密度为3.8%～9.7%.多巴胺的平均回收率为107.3%、去甲肾上腺素为89.4%、肾上腺素为80.1%,其余4种物质为89.2%～95.7%.②2例嗜铬细胞瘤患者24 h尿中儿茶酚胺类物质的含量显著高于正常人,其中1例以肾上腺素明显增高为主,另1例以去甲肾上腺素明显增高为主,均高于正常人10倍左右.③帕金森病大鼠健侧尾壳核内多巴胺及其代谢产物多巴柯、高香草酸含量均在正常范围内,损毁侧尾壳核内多巴胺、5-羟色胺、多巴柯及高香草酸均显著降低,差异有显著性意义（P＜0.01）;5-羟色胺及其代谢产物5-羟吲哚乙酸含量无明显改变.④帕金森病大鼠健侧尾壳核内多巴胺更新率（多巴柯/多巴胺和高香草酸/多巴胺的比值）升高;损毁侧尾壳核内多巴胺更新速度也加快,与正常对照组比较差异均有显著性意义（P＜0.05）.结论：高效液相色谱法能满足对大白鼠帕金森病模型的研究和对嗜铬细胞瘤患者实验诊断的需要.     

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.     

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.     

Prospects for the control of neglected tropical diseases by mass drug administration

The prospects for the control of neglected tropical diseases, including soil-transmitted helminthiasis, shistosomiasis, lymphatic filariasis, onchocerciasis and trachoma, through mass drug administration, are exemplified by the elimination of the trachoma as a public-health problem in Morocco. In spite of this and other striking successes, mass drug administration programs are faced with major challenges resulting from suboptimal coverage and lack of efficacy. At current suboptimal coverage rates, programs may need prolongation for an extended period, increasing costs and undermining sustainability. Community participation through health education and information appears to be crucial to improve coverage and to achieve sustainability. Implementation of complementary measures, such as vector control, improved hygiene and environmental sanitation, are important to further control transmission and to prevent re-emergence of the infection and, again, may only be achieved effectively through community-based initiatives. To reduce costs and to relieve pressure on the health system, combining neglected tropical disease programs in areas where diseases coexist and integration with existing control programs for malaria, tuberculosis and HIV/AIDS is advocated. The risk of developing drug resistance is of particular concern in view of the lack of alternative drugs, and reduced treatment efficacy due to emerging resistance is evident for the soil-transmitted helminths and onchocerciasis. Given the risk for the development of drug resistance and the need for a high degree of participation, close attention should be paid to the monitoring of the coverage and efficacy of the different program components.     

Bacterial meningitis: an update of new treatment options

The outcome of bacterial meningitis critically depends on the rapid initiation of bactericidal antibiotic therapy and adequate management of septic shock. In community-acquired meningitis, the choice of an optimum initial empirical antibiotic regimen depends on the regional resistance patterns. Pathogens resistant to antibacterials prevail in nosocomial bacterial meningitis. Dexamethasone is recommended as adjunctive therapy for community-acquired meningitis in developed countries. In comatose patients, aggressive measures to lower intracranial pressure <20 mmHg (in particular, external ventriculostomy, osmotherapy and temporary hyperventilation) were effective in a case–control study. Although many experimental approaches were protective in animal models, none of them has been proven effective in patients. Antibiotics, which are bactericidal but do not lyse bacteria, and inhibitors of matrix metalloproteinases or complement factor C5 appear the most promising therapeutic options. At present, vaccination is the most efficient method to reduce disease burden. Palmitoylethanolamide appears promising to enhance the resistance of the brain to infections.     

Imaging choices in occult hip fracture

Background: Hip fracture is a common injury, with an incidence rate of > 250,000 per year in the United States. Diagnosis is particularly important due to the high dependence on the integrity of the hip in the daily life of most people. Objectives: In this article we review the literature focused on hip fracture detection and discuss advantages and limitations of each major imaging modality. Discussion: Plain radiographs are usually sufficient for diagnosis as they are at least 90% sensitive for hip fracture. However, in the 3–4% of Emergency Department (ED) patients having hip X-ray studies who harbor an occult hip fracture, the Emergency Physician must choose among several methods, each with intrinsic limitations, for further evaluation. These methods include computed tomography, scintigraphy, and magnetic resonance imaging. Conclusion: We present an evidence-based algorithm for the evaluation of a patient suspected to have an occult hip fracture in the ED. Also outlined are future directions for research to distinguish more effective techniques for identifying occult hip fractures.     

Systematic Reviews Published in the April 2012 Issue of the Cochrane Library

ABSTRACT

The Cochrane Library of Systematic Reviews is published quarterly as a DVD but monthly online. The April 2012 issue (second DVD for 2012) contains 5045 complete reviews, 2182 protocols for reviews in production, and 17,084 short summaries of systematic reviews published in the general medical literature. In addition, there are citations of 674,000 randomized controlled trials, and 15,400 cited papers in the Cochrane methodology register. The health technology assessment database contains just over 11,000 citations. One hundred and seventeen new reviews have been published in the last 3 months of which 12 have potential relevance for practitioners in pain and palliative medicine. The impact factor of the Cochrane Library stands at 6.186. Readers are encouraged to access the full report for any articles of interest as only a brief commentary is provided.     

Platelets,thrombosis, coagulation,and atherosclerosis

When I first got the invitation to join a medical delegation going to Moldova, I thought for a moment that our destination was the fictional country in the old Marx Brothers movie Duck Soup. On further checking, it turns out that entertaining place was called Freedonia. I now know that Moldova is indeed a real country, bordered on the west by Romania and on the other three sides by the Ukraine. It is a proud country, rich with traditions, and its people are warm, giving, eager to learn ways to improve their healthcare system, and deeply appreciative of our attempts to help them in the task.     

Advance Care Planning

Predictors of patient wishes and influence of family and clinicians are discussed. Research findings on patient decision-making relating to preferences in end-of-life care are described. Advance directives and durable powers of attorney are defined and differentiated. Most patients have not participated in advance care planning and the need for more effective planning is documented. Appropriate times for discussions of such planning are described. Scenarios discussed include terminal cancer, chronic obstructive pulmonary disease, AIDS, stroke, and dementia. Patient satisfaction is discussed, as is a structured process for discussions about patient preferences. Results of patient responses to hypothetical scenarios are described. Invasiveness of interventions, prognosis and other factors that favor or discourage patient preferences for treatment are discussed. Findings resulting from research funded by the Agency for Healthcare Research and Quality (AHRQ) are discussed. This research can help providers offer end-of-life care based on preferences held by the majority of patients under similar circumstances.     

Spiritual Dimensions of Nursing Practice. edited by V. B. Carson

The Cochrane Library of Systematic Reviewsis published quarterly. Issue one for 2004 of the library was published in February 2004. This issue contains 3,329 reviews and protocols of which 1,921 are fully published reviews. The trials database now stands at over 400,000 records with an additional 4,427 one-page summaries of non-Cochrane reviews in the NHS database of reviews of effectiveness (DARE). This version of the library contains the results of an extensive search for RCTs on EMBASE. The latest library contains 84 new reviews, seven are considered relevant to practitioners in pain and palliative care. References are published in the same format as the citation for Cochrane reviews.     

Évaluation,par l’imagerie ultrasonore,du recrutement musculaire,de la force motrice et de la détente verticale du quadriceps après électromyostimulation (cas de 15 étudiants de l’INJS)

The increase of strength seems to be an element of performance in some sports.