降钙素原生物素-链霉亲和素酶联免疫吸附分析法的建立

目的建立生物素一链霉亲和素酶联免疫吸附分析（biotin．streptavidinamplifiedELISA,BA．ELISA）法半定量检测人血清中降钙素原含量。方法通过双抗体夹心法,结合生物素一链霉亲和素系统的放大作用,建立降钙素原的半定量检测方法。并对该试剂的各项性能指标进行评价。结果该方法的最低检测限为0．12mg／mE,线性范围为0．12～20ng／mL,回收率为98．59％。分析内和分析间变异系数分别为4．9％～16．8％和7．1％～14．9％。与降钙素原结构类似物及血清中常见的干扰物质特异性良好。通过对73例血清样本进行检测分析,当阈值为0．5ng／mL时,敏感性和特异性分别为100％和96．0％。结论本研究建立的降钙素原生物素一链霉亲和素酶联免疫吸附分析法具有灵敏、简便、准确等特点,具有良好的应用前景。     

人血浆β淀粉样蛋白42ELISA的建立及其临床初步应用

目的建立人血浆中β淀粉样蛋白42（Aβ42）酶联免疫吸附试验（ELISA）,并初步观察其对诊断阿尔茨海默病（AD）的应用价值。方法用鼠抗Aβ42片段（1～28氨基酸残基）单克隆抗体包被微孔板,用生物素标记的兔抗Aβ42片段（40～42氨基酸残基）为标记抗体,应用生物素-亲和素系统（ABS）进行放大,采用棋盘滴定方法确定最佳实验条件,用Aβ42标准品建立ELISA标准曲线,分别检测不同痴呆程度AD患者及健康人血浆中Aβ水平。结果本研究建立的方法测定范围为20—500pg／mL,最低检出量20pg／mL,批内和批间变异系数（c叻分别为3．8％和7．9％。轻度AD患者血浆中AB42水平为（146．4±9．7）pg／mL,明显高于健康对照组[（95．I±7．2）pg／mL],中晚期AD患者血浆Aβ42浓度明显下降[（99．4±17．6）pg／mL],其水平降至与健康对照组差异无统计学意义。结论建立的人血浆Aβ42ELISA可作为AD早期诊断的有效方法,可在临床推广应用。     

血清肌红蛋白光激化学发光免疫测定法的建立

目的采用光激化学发光免疫测定技术（LICI）建立定量检测肌红蛋白（Mb）的方法。方法使用针对Mb不同表位的2株单克隆抗体，一株包被发光微粒，另一株进行生物素化，与包被有链霉亲合素的感光微粒共同构成检测试剂，优化检测条件并评价检测性能。结果该方法的灵敏度为1．36ng／ml，在33．8～1521．0ng／ml的范围内线性良好。批内和日间变异系数分别为3．05％～4．41％和5．80％～6．56％。在血红蛋白浓度〈2．6g／L，总胆红素浓度〈342μmol／L、三酰甘油浓度〈11．3mmol／L时的干扰率无临床意义。与Roche Elecsys电化学发光法有较好的相关性（r=0．9971）。结论血清MbLICI的建立有助于进一步对检测试剂盒的研制。     

生物素-链霉亲和素酶免疫法PSA测定的评价

目的 评价生物素-链霉亲和素酶免疫分析法(BSA-ELISA)测定前列腺特异性抗原(PSA)并与化学，发光法(CLIA)进行比较。方法根据EP9-A文件要求采集数据，并以相应软件分析这两种方法的相关性；同时对BSA-ELISA的测定范围，变异系数(CV)，最小检测限进行测定。结果两种方法测定的结果具有较好的相关性。回归方程Y=0．985X-0．131，相关系数(r)=0．997。PSA的批内平均CV为2．21％，批间平均CV为3．08％。测定范围为1．50～85．00μg／L。最小检测限为0．30μg／L。结论采用全自动酶免疫分析仪的BSA-ELISA与CLIA自动分析仪测定PSA的相关性好，结果稳定，偏差小，可以在临床上常规应用。     

生物素—链霉亲和素免疫学法测定地高辛直接包被方法的研究

笔者曾经成功地应用德国ＢｏｅｈｒｉｎｇｅｒＭａｎｈｅｉｍ公司生产的现成的链霉亲和素包被管建立了生物素－链霉亲和素免疫学潮测定地高辛的反应模型。本文用国产的链霉亲和素自行包被，采用的是一种直接包被方法，即将链霉亲和素溶于去离子水，冰箱过液包被２４ｈ，并应用到地高辛的临床检测中，其灵敏度为０．０７８１μｇ／Ｌ，最低检测限为０．１９５２μｇ／Ｌ，测定三份低、中、高浓度血清标本，批内变异系数分别为８．７％     

间接包被ELISA法检测血清游离β-HCG

目的 研制生物素-亲合素间接包被ELISA法定量检测血清游离β-HCG试剂盒。 方法 用生物素化牛血清清蛋白和链霉亲合素包被微孔反应板,生物素化抗游离β-HCG抗体和酶标记抗游离β-HCG抗体检测游离β-HCG,评价试剂盒相关技术参数。 结果 本试剂盒具有较好的稳定性,灵敏度0.1 ng/ml,回收率95.9％~102.5％,批内CV 7.5％~11.1％,批间CV12.5%~18.1%,与FSH、TSH、LH和α-HCG的交叉反应率均<0.1％,检测范围0.1~500 ng/ml,与美国DRG公司试剂盒的相关系数r2=0.789 2,P<0.05。 结论 本方法研制的试剂盒灵敏度高,特异性好,操作方法简便,适于临床常规应用。     

B型钠尿肽酶免疫检测的研究

目的 采用酶免疫分析技术,建立B型钠尿肽(BNP)酶联免疫吸附方法(ELISA).方法 使用纯化的对一抗(兔抗BNP抗体)特异的二抗(羊抗兔抗体)包被微孔板,然后再加入一抗.制成固相抗体.检测中往包被抗体的微孔中依次加入BNP抗原或检测样品、生物素化抗人BNP抗体、HRP标记IgG,经过彻底洗涤后加底物液(OPD)显色,用酶标仪在492nm波长下测定吸光度(A值),计算样品浓度.结果 该法测定BNP的灵敏度为15 pg/ml,批内和批间CV分别为5.3±1.3%和7.1±1.5%,平均回收率为95.9±1.4%,线性范围为25～4 000 pg/ml.该方法测定正常组(n=20)及心衰组(n=25)血浆BNP分别为56.8±25.9(0～132 pg/ml)及462.3±158.7(163.4～697.8 pg/ml),而免疫放射法测定值分别为59.3±27.2(0～140 pg/ml)及470.1±162.5(165.8～708.6 pg/ml),两种方法所测结果高度相关,相关系数r=0.92,P<0.01.结论 应用酶免疫技术初步建立测定BNP的ELISA法,检测范围宽,重复性好,具有较高灵敏性及特异性.     

BAS-TRFIA法检测抗环瓜氨酸肽抗体方法学的建立

目的利用生物素-链霉亲和素放大系统建立一种可快速、灵敏地定量检测患者血清中抗环瓜氨酸肽(anticyclic citrullinated peptide,CCP)抗体的时间分辨荧光免疫分析法(time-resolved fluoroimmunoassay,TRFIA)。方法将链霉亲和素同铕标记二乙烯三胺五乙酸酐结合制备铕标记链霉亲和素衍生物;生物素同兔抗人IgG抗体结合制备生物素化IgG抗体,后者在免疫检测系统中可联结铕标记亲和素和抗CCP抗体从而形成复合物。最后通过测量Eu3+-链霉亲和素在615nm的荧光值计算血清抗CCP抗体水平。结果该方法具有更宽的线性范围,其线性范围为0.58-9463U/ml,而应用ELISA试剂盒检测线性范围只有18.48-591.4U/ml。此外,该方法的抗CCP抗体检测限可达0.5U/mL。回收率为96.45-104.63%。用BAS-TRFIA和ELISA法测得的值具有很好的相关性(R2=0.892 7)。结论本研究数据表明我们的建立的BAS-TRFIA法在测定抗CCP抗体上较传统ELISA试剂盒有很大改进,为RA的诊断和治疗提供了一个更为理想的免疫方法学选择。     

丙型肝炎病毒抗体桥式双抗原夹心时间分辨荧光免疫分析法的建立

目的建立丙型肝炎病毒抗体桥式双抗原夹心时间分辨荧光免疫分析法。方法采用丙型肝炎病毒基因重组多表位嵌合蛋白作为包被抗原,生物素标记抗原作为桥接抗原,链霉亲和素标记铕作为示踪物,配制以β-萘甲酰三氟丙酮为主要成分的增强液,自建方法的校准品以国家标准血清物质GBW（E）090047为溯源,应用桥式双抗原夹心法建立丙型肝炎病毒抗体时间分辨荧光免疫分析法。结果自建方法在0．031—4．00NCU／ml内,相关系数可达0．99;分析内和分析间变异系数均小于10％;灵敏度可达0．016NCU／ml;校准品的效价比在0．90～1．10,平均回收率为101．47％;与相关疾病的抗原或抗体无明显交叉反应,其表观浓度值均小于0．03NCU／ml;参考值为0．05NCU／ml;符合国家检定标准的要求;37℃恒温箱烘烤7d后检测性能无明显改变;与同类方法学比对符合性好。结论自建方法精密度好,灵敏度高,特异性强,准确性好,线性范围宽,符合率高,能满足临床检测需要。     

急性心肌梗死患者血浆心肌营养素-1水平及对预后预测的价值

【目的】探讨急性心肌梗死（AMI）患者血浆心肌营养素-1（CT-1）水平的变化及其对预后预测的价值。【方法】检测86例AMI患者入院时血浆CT-1及心肌肌钙蛋白I（cTnI）的浓度,另选24名健康体检者为对照组,根据CT—l浓度将AMI患者分为CT-1〈300pg／mL组（n=52）和CT-1〉300pg／mL组（n=34）,记录患者1个月内发生的心血管事件（心血管意外死亡、心力衰竭、再发心绞痛或再发心肌梗死）,观察AMI后CT-1水平的变化、CT-1与cTnI的相关性及其对AMI患者的预后价值。【结果】AMI患者血浆CT-1浓度明显高于对照组（P〈0．01）;血浆CT-1水平与cTnI成正相关（r=0．761,P〈0．01）;CT-1〉300pg／mL组心血管事件发生率明显高于CT-1〈300pg／mL组（P〈0．01）;多变量回归分析,CT-1是预测AMI患者发生近期心血管事件的有效指标（RR：1．753,95％CI为1．112～3．438,P=0．012）。【结论】血浆CT-1水平能够反映AMI患者心肌坏死程度,也是预测AMI患者发生近期心血管事件的有效指标。     

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.     

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.     

Clinical Pharmacokinetics of Morphine

Morphine, the most widely used mu-opioid analgesic for acute and chronic pain, is the standard against which new analgesics are measured. A thorough understanding of the pharmacokinetics of morphine is required in order to safely and effectively use this analgesic in a wide variety of patients with different levels of organ function. A MEDLINE search was conducted to identify literature published between 1966 and January 2002 relevant to the pharmacokinetics of morphine. These publications were reviewed and the literature summarized regarding unique and clinically important elements of morphine disposition relative to its parenteral administration (including intravenous, intramuscular, subcutaneous, epidural and intrathecal administration), absorption profile (immediate release, controlled release, and sublingual/buccal, and rectal administration), distribution, and its metabolism/ excretion. Special populations, including infants, elderly, and those with renal/liver failure, have a unique morphine pharmacokinetic profile that must be taken into account in order to maximize analgesic efficacy and reduce the risk of adverse events.     

手转胎头术失败的原因与分娩结局临床分析

目的 探讨手转胎头术失败的原因与分娩结局.方法 选择2008年1月至2010年12月于我院住院分娩的持续性枕横位、枕后位产妇198例,根据行手转胎头术后结果分为成功组126例、失败组72例.比较两组分娩结局,对比分析失败原因.结果 失败组胎儿体质量≥3500 g的发生率[76.4％(55/72)]明显高于成功组[31.7％(40/126)],差异有统计学意义(x2=30.177,P=0.001)、失败组宫缩乏力发生率[58.3％(42/72)]高于成功组[38.1％ (48/126)],差异有统计学意义(x2=7.569,P=0.006)、失败组骨盆临界或轻度狭窄发生率[38.9％ (28/72)]高于成功组[23.8％(30/126)],差异有统计学意义(x2 =5.030,P=0.002)、失败组手转胎头时机不当(宫口开大＜6 cm、胎头位于坐骨棘上及宫口开大8～10 cm、胎头位于坐骨棘下≥2 cm)发生率[61.1％(44/72)]高于成功组[38.9％(49/126)],差异有统计学意义(x2=9.084,P=0.003).失败组母儿并发症(产后出血、产褥病率、胎儿窘迫、新生儿窒息)发生率高于成功组(x2 =9.586,P=0.002、x2=9.334,P=0.002、x2=5.910,P=0.015、x2=5.240,P=0.022)、失败组剖宫产发生率[72.2％(52/72)]明显高于成功组[34.1 ％(43/126),x2=26.641,P=0.001)].结论 手转胎头术能使难产变顺产,降低剖宫产率,减少母儿并发症,但须积极预防、处理导致手转胎头术失败的原因,对矫正失败后继续矫正及试产应慎重.     

Evidence-Based Pain Management and Palliative Care in Issue Four for 2009 of The Cochrane Library

ABSTRACT

The Cochrane Library of Systematic Reviews is published quarterly. Issue 4 for 2009 contains 4027 complete reviews, 1906 protocols for reviews in production, and 11447 one-page summaries of systematic reviews published in the general medical literature. In addition, there are citations of 600,000 randomized controlled trials, and 12,200 cited papers in the Cochrane methodology register. The health technology assessment database contains over 7500 citations. This edition of the Library contains 90 new reviews, of which 19 have potential relevance for practitioners in pain and palliative medicine.     

Wie zuverlässig sind Angaben zu nichtentzündlichen ausgedehnten Schmerzen?

ZusammenfassungFragestellung Es wurde geprüft, wie sich der Differenziertheitsgrad zweier Schmerzmessmethoden auf Angaben zur Ausgedehntheit klinischer Schmerzen auswirkt. Zugleich wurde der Referenzzeitraum variiert, über den die Patienten berichten sollten.Methode Erfasst wurde der Einfluss zu Lasten der Befragungsdifferenziertheit durch den Vergleich zweier Körperschema-Bildvorlagen. Drei Referenzzeiträume (Schmerz aktuell, letzte Woche, letztes halbes Jahr) wurden vorgegeben.Ergebnisse Patienten mit ausgedehnten Schmerzen gaben bei differenzierter Befragung um so mehr Schmerzen an, je weiter die Schmerzen zurück lagen und je größer der Berichtszeitraum war. Patienten mit gelenknahen Schmerzen gaben bei hoch differenzierter Befragung weniger ausgedehnte Schmerzen in der Vergangenheit an als bei globaler Einschätzung. Patienten mit Rückenschmerzen berichteten bei differenzierter Befragung zum aktuellen Schmerz über weniger ausgedehnte Schmerzen als bei globaler Befragung.Schlussfolgerung Die Angaben zur Schmerzausdehnung variieren vor allem bei Patienten mit ausgedehnten Schmerzen in Abhängigkeit von der Differenziertheit der Befragung. In diesen Fällen ist die Wahrscheinlichkeit erhöht, dass sich die Beschwerdesymptomatik zumindest teilweise erst in der Reaktion auf die situativen Befragungsbedingungen konstituiert und daher nicht auf andere Befragungsbedingungen generalisiert werden kann.