流式细胞术与微量淋巴细胞毒法检测HLA-B27的比对

目的:评价实时流式细胞术(FCM)和微量淋巴细胞毒法(MCLT)检测HLA-B27对强直性脊柱炎(AS)的临床诊断价值.方法:采用FCM和MCLT同步检测131例AS患者外周血标本的HLA-B27的表达.结果:HLA-B27阳性率在AS组高达96.47%,AS组与非AS组和健康体检组差异均有统计学意义(P<0.01).两种方法比较,具有高度一致性,Kappa值>0.70.FCM的准确性达96.3%,特异性、灵敏度分别为96.7%和96.5%.与MCLT技术相比,减少了假阳性.结论:FCM法对HLA-B27测定特异性、灵敏度都明显超过MCLT法,是目前较为理想的方法.     

试纸法快速测定全血血糖与已糖激酶法测定血清糖结果比较

目的：了解试纸法快速测定血糖结果与参考方法测定结果的符合程度。方法：用Surestep Plus稳步倍加型血糖仪快速测定血糖与Beckman Synchron CXΔ测定血清糖作参考对照。结果：6份毛细血管(末梢)全血，静脉全血，静脉血清用试纸法快速测定血糖与静脉血清已糖激酶法测定血糖结果之间差异无显意义。结论：试纸法快速测定全血血糖结果与参考方法测定血糖结果高度正相关r=0．981。     

密度梯度离心联合上皮细胞黏附分子免疫磁珠富集法检测卵巢癌循环肿瘤细胞及其与肿瘤复发转移的相关性

目的检测卵巢癌患者外周血中循环肿瘤细胞(CTCs)与肿瘤复发转移的相关性。方法采集50例卵巢癌患者的外周血,应用密度梯度离心联合上皮细胞黏附分子免疫磁珠富集法检测外周血中的CTCs的数目,并比较其与患者临床特征之间的关系。结果 50例卵巢癌患者中32例为CTCs阳性,CTCs阳性与远处转移(P<0.05)和复发密切相关(P<0.05),而与性别、年龄、肿瘤部位、TNM分期、肿瘤分化程度及淋巴转移无关(P>0.05)。结论 CTCs可以预测卵巢癌远处转移和复发。     

LC-MS/MS法和mFPIA法测定全血环孢素A浓度的相关性比较

目的:对高效液相色谱电喷雾串联质谱(LC-MS/MS)法和单克隆荧光偏振免疫(mFPIA)法测定肾移植术后稳定期患者不同时间点的全血环孢素A(CsA)浓度进行相关性比较。方法:分别采用LC-MS/MS法和mFPIA法测定患者全血CsA浓度,对测定结果进行双侧配对t检验比较差异性,用Passing-Bablok回归分析法比较不同采血时间点2种方法测定结果的相关性。结果:2种方法测定值之间具有显著性差异;服药后0、3.0、4.0h时2法测定值的相关性最好,0.5、0.75、12.0h相关性最差。结论:对CsA血药浓度监测结果的分析需考虑监测方法以及不同采血时间点的影响。     

比较用凝集素微量离心柱法和酶联免疫吸附法检测甲胎蛋白异质体对原发性肝癌的诊断价值北大核心CSCD

目的比较用凝集素微量离心柱法(ACSC)和酶联免疫吸附法(ELISA)检测甲胎蛋白异质体对原发性肝癌的诊断价值。方法选取2013年7月至2015年12月于本院就诊的原发性肝癌(HCC)患者85例,慢性肝炎(CH)与肝硬化(LC)患者38例;用ELISA法检测甲胎蛋白异质体(AFP-L3)浓度,用ACSC法检测AFP(甲胎蛋白)-L3/AFP%,比较2种检测办法对良恶性肝病的鉴别诊断价值。结果 ACSC法在HCC中AFP-L3/AFP%≥10%阳性率为84.70%(72例/85例),ELISA法在HCC中AFP-L3>37.89 ng·m L^(-1)的阳性率为90.59%(77例/85例)。2种方法检测CH与LC的阳性率分别为13.16%(5例/38例)、10.53%(4例/38例),两者比较差异有统计学意义(P<0.05)。在2种方法对良恶性肝病鉴别诊断性能评价的ROC曲线中,AUC值分别为0.89,0.82(均P<0.01),95%可信区间分别为0.81~0.98,0.74~0.90。2种方法诊断HCC的敏感性分别为84.71%(72例/85例),88.24%(75例/85例);特异性分别为86.84%(33例/38例),89.47%(34例/38例);准确性分别为85.36%(105例/123例),88.62%(109例/123例),这三者比较差异无统计学意义(均P>0.05)。结论 2种方法诊断HCC的敏感性、特异性、准确性类似,但ELISA法具有操作简单、价格低廉等优点,更适合在临床上推广应用。     

2型糖尿病尿微量白蛋白与大血管病变的相关关系研究

近年来 ,尿微量白蛋白与糖尿病大血管病变的关系越来越受到重视[1,2 ] 。本文通过分析 2 2 6例 2型糖尿病病人的尿白蛋白排泄率 (UAER) ,目的在阐明其与大血管病变及相关因素之间的关系 ,现报告如下。1　对象与方法1 1　对象　共收集住院及专科门诊的 2型糖尿病病人 2 2 6例 ,均符合WHO(1998年 )糖尿病诊断标准 ,按尿白蛋白排泄率将病人分三组 ,Ⅰ组 (正常白蛋白尿组 )为 <2 0 μg/min ,Ⅱ组 (微量白蛋白尿组 ) >2 0～ 2 0 0 μg/min ,Ⅲ组 (大量白蛋白尿组 )为 >2 0 0 μg/min ,非糖尿病组 30例为正常对照组。各组例数及临床情况如见表 …     

HPLC—MS法和mFPIA法测定全血环孢霉素A及其代谢物的比较

目的:对测定全血环孢霉素 A 的 HPLC-MS 法和单克隆荧光偏振免疫分析法(mFPIA 法)进行评价。方法:建立HPLC-MS 测定全血环孢霉素 A 及其代谢物的方法,采用多元线性回归对 HPLC-MS 法和 mFPIA 法测定的结果进行比较。结果:当患者肝、肾功能异常时,mFPIA 法测定结果同环孢霉素 A 及主要代谢物浓度呈线性相关,易导致检测结果偏高。结论:当患者肝、肾功能异常时,mFPIA 法不适宜作为环孢霉素 A 的临床监测方法。     

Neo-Sensitab纸片扩散法与NCCL SM27-A微量稀释法测定97株酵母对抗真菌药物敏感性的比较

目的比较Neo-Sensitab纸片扩散法和NCCLS微量稀释法的一致性,评价该商品化方案的稳定性。方法同时采用NCCLSM27-A微量稀释法和Neo-Sensitab纸片扩散法测定临床分离的97株酵母对两性霉素B(AMB)、氟康唑(FCZ)、5-氟胞嘧啶(5-FC)、酮康唑(KCZ)和伊曲康唑(ITC)等五种常用抗真菌药物的敏感性。结果两种方法的药敏结果一致率分别为AMB100%,FCZ94.8%,5-FC93.8%,KCZ75.3%和ITC84.5%。结论纸片扩散法和微量稀释法药敏测试结果一致率较高,以FCZ、AMB和5-FC最佳,该方法检测敏感株的准确率较高,而在区分耐药株和中介株方面有所不足。     

激光散射法和电阻抗法在全血血小板计数中的比较

目的探讨激光散射法和电阻抗法在全血血小板（PⅡ）计数的准确性。方法运用SYSTEMXT-2000i全自动血细胞分析仪同时检测电阻抗法血小板和激光散射法血小板,按电阻抗法将检测结果分为PLT〈50×10。·L～、（50—100）×10^9·L^-1、（100—300）×10^9·L^-1、〉300×10^9·L^-1共4组,将两种检测方法结果按误差分为：误差在10％以内标本212例,误差在10％以上标本150例,采用10g.L^-1草酸铵溶液稀释人工显微镜计数血小板,评价激光散射法和电阻抗法的准确性,以手工计数为准。结果电阻抗法和激光散射法计数血小板,在误差10％以内,两种方法在四组标本中差异均无显著性（P〉0．05）;在误差10％以上时,当PLT〈100×10^9·L^-1和〉300×10^9-L^-1。时,电阻抗法与显微镜法比较,差异均有显著性（P〈0．05）,而激光法与显微镜法比较,差异没有显著性（P〉0．05）;所有组别电阻抗法与显微镜法比较,差异均有显著性（P〈0．05）,而激光法与显微镜法比较差异均无显著性（P〉0．05）。结论电阻抗法计数血小板仍然是目前最经济有效的方法。对于低值和高值血小板检测,激光散射法比电阻法准确性高,人工显微镜复检是最客观准确的方法之一。     

测定环孢素全血浓度的2种方法对比及临床应用

目的对比荧光偏振免疫法(FPIA)和均相酶扩大免疫分析法(EMIT)2种测定方法监测环孢素(CsA)血药浓度的差异。方法收集98例次环孢素全血样品,同日同一患者的全血样品分别采用FPIA和EMIT法测定血药浓度。结果 EMIT和FPIA测定结果相关系数r为0.9795,EMIT法较FPIA法平均低(11±12)%(P<0.05,n=98),其中CsA血药谷值浓度(c_0)EMIT法较FPIA法平均低(14±9)%(P<0.05,n=69)。CsA血药峰值浓度(c_2)EMIT法较FPIA法平均低(2±5)%(P>0.05,n=29)。结论 2种不同方法测定全血中CsA血药浓度具有显著差异,EMIT法较FPIA法具有较高的特异性。     

Semiautomated high-performance liquid chromatographic method for the determination of benzodiazepines in whole blood

A semiautomated method for the determination of five frequently prescribed benzodiazepines (BZD) (clonazepam, diazepam, flunitrazepam, midazolam, and oxazepam) in whole blood samples by reversed-phase high-performance liquid chromatography following simple online enrichment and clean-up on a short precolumn is described. After precipitation of protein and red cells with a mixture of organic solvents (methanol/acetonitrile, 50:50), the aliquot is centrifuged and the organic upper phase evaporated under a gentle stream of nitrogen. The residue is reconstituted by adding 500 microL of a mixture of phosphate buffer (20mM, pH 2.2) and acetonitrile (70:30, v/v). The sample is then directly introduced into the column-switching column. The precolumn is first washed with phosphate buffer at pH 7.2. Compounds retained on the precolumn are then eluted in the back-flush mode and separated on a C8 semi-microcolumn (Lichrospher select B, 125 x 3 mm). The BZD studied are determined by a diode-array detector at 254 nm. The method shows excellent linearity between 25 and 1,000 ng/mL for clonazepam, flunitrazepam, and midazolam and between 25 and 5,000 ng/mL for diazepam and oxazepam. The recoveries are around 80% for clonazepam and oxazepam and around 90% for the three others. Coefficients of variation for between-day and within-day assays are < 15% for low concentrations close to the limit of quantitation and < 5% for high concentrations.     

用LC/LC-UV系统建立全血中环孢霉素A自动化测定方法

目的建立自动化LC/LC-UV系统建立全血中环孢霉素A测定方法。方法 LC1色谱柱FRO-XB CN（20mm×4.6mm,10μm,ANAX）,流动相为水-乙腈=35∶65（v/v）,流速为1.0mL.min-1,中央处理体系温度为40℃,LC2系统中分析色谱柱为AC-CN（200mm×4.6mm,5μm,ANAX）,流动相为乙腈-水,采用梯度系统,0～15min流动相中乙腈比例从40%直线上升到53%,流速为0.7mL.min-1。转移辅助流速为0.7mL.min-1。目标物转移窗口为2.20～3.20min;定量环为500μL,样品用乙腈预处理。结果环孢霉素A在0.026～1.010μg.mL-1与峰面积线性关系良好,平均相关系数为0.999 1,萃取回收率在101.0%～103.0%,最低定量浓度为0.013μg.mL-1。日内RSD〈5%（n=6）,日间RSD〈5%（n=18）,准确度在100.3%～102.6%。结论用自动化LC/LC-UV系统建立的环孢霉素A测定方法人为干预步骤少、准确性与精密度良好,方法开发、运行及控制自动化程度高,适合于环孢霉素A血药浓度测定及动力学研究。     

A rapid, reliable high-performance liquid chromatographic micromethod for the measurement of cyclosporine in whole blood

The initiation of a liver transplant program in a pediatric hospital prompted the development of a high-performance liquid chromatography method for monitoring cyclosporine, which requires minimal amounts of blood and produces fast, reliable results. A protein-free filtrate is prepared by mixing 250 microliters of whole blood with 750 microliters of organic solvent (acetonitrile and methanol in the ratio of 9:1) containing the internal standard cyclosporine D. After centrifugation the supernatant is subjected to a quick clean-up using two types of disposable cartridges (C18 and silica Bond Elut), and the eluate is dried, reconstituted in equal parts of acetonitrile and water, and subjected to a final heptane wash to remove late eluting peaks. Chromatography is performed at 75 degrees C using a supelcosil C18 column (15 cm X 4.6 mm) with 3 microns packing and a mobile phase of 77/23 of acetonitrile/phosphate buffer at pH 2.5. Flow rate is 0.6 ml/min, providing a chromatography time of 10 min. Absorbance is measured at 214 nm at a sensitivity setting of 0.01 absorbance units full scale. Recovery for cyclosporine A is between 92 and 104%, and the lower level of sensitivity is 15 micrograms/L. Between-day precision provided coefficient of variation values less than 5% for a control serum of 150 micrograms/L. The method is linear to 3,000 micrograms/L. No interfering substances have been found. The method has proved to be consistent, reliable, and simple enough to be performed by all staff members. Column life for a system used daily is at least 3 months.     

流式细胞仪高通量药物筛选技术的建立和应用

目的建立流式细胞仪高通量药物筛选技术,并用于筛选逆转肿瘤细胞多药耐药性的药物。方法使用流式细胞仪高通量附件,建立高通量药物筛选技术;以罗丹明123作为荧光指示剂,利用耐药HR-20活细胞筛选逆转耐药性的药物。结果通过阳性药物,确定了逆转耐药性药物筛选模型的Z′因子为0.88。应用此模型筛选8 000多种化合物。结论流式细胞仪高通量药物筛选技术是一种精确度较高、稳定性较好、有效的高通量药物筛选方法。     

Development and validation of a GC/MS method for the determination of tadalafil in whole blood

Tadalafil is a phosphodiesterase type 5 (PDE-5) inhibitor and it is used in the treatment of pulmonary arterial hypertension and erectile dysfunction. A sensitive and specific method is described for the determination of tadalafil in whole blood. Tadalafil and its internal standard (protriptyline) were isolated from the matrix by solid phase extraction, and were analyzed by gas chromatography/mass spectrometry (GC/MS) after derivatization by N,O-bis(trimethylsilyl)-trifluoracetamide (BSTFA) with 1% trimethylchlorsilane (TMCS). Limits of detection and quantification for tadalafil were 0.70 and 2.00 μg/L, respectively. The calibration curve was linear between 2.00 and 500.0 μg/L, with a correlation coefficient higher than 0.991. The values obtained for intra- and inter-day accuracy was found to be between −10.5 to 8.5% and −4.2 to 4.5%, respectively, while intra- and inter-day precision were less than 8.4 and 11.2%, correspondingly. Absolute recovery was determined at three concentration levels and ranged from 92.1 to 98.9%. The proposed method is the first fully validated GC/MS method for the determination of tadalafil in whole blood and it can be routinely applied by toxicological laboratories, for pharmacokinetic studies, for therapeutic drug level monitoring or for the investigation of related forensic cases.     

Clinical use of the determination of serotonin in whole blood

Whole blood serotonin (5-hydroxytryptamine, 5-HT) was assayed, and factors possibly influencing 5-HT content were investigated in healthy controls. No significant circadian rhythm or effect of dexamethasone or meals was observed. After use of fluvoxamine, a specific 5-HT reuptake inhibitor, the whole-blood 5-HT concentration of patients was strongly reduced. After treatment with tranylcypromine, an unspecific monoamine oxidase inhibitor (MAOI), the 5-HT content was increased. Determination of whole blood 5-HT in patients treated with fluvoxamine presents a measure of drug compliance. Furthermore, the method may have clinical potential for finding an adequate dose when a 5-HT reuptake inhibitor or an MAOI is used.     

Automated solid-phase extraction method for the determination of piperaquine in whole blood by rapid liquid chromatography

A bioanalytic method for the determination of piperaquine in whole blood by solid-phase extraction and rapid liquid chromatography has been developed and validated. Whole blood was hemolyzed with deionized water, and an internal standard was added to the samples before they were loaded onto a PRS cation-exchange solid-phase extraction column. Piperaquine and internal standard were analyzed by liquid chromatography on a Chromolith Performance (100 x 4.6 mm) column with mobile phase acetonitrile:phosphate buffer, I = 0.1, pH 2.5 (8:92, vol/vol), flow rate 4 mL x min-1, and UV detection at 345 nm. The intraassay precision for whole blood was 3.2% at 3.00 microM and 12.3% at 0.100 microM. The interassay precision for whole blood was 1.8% at 3.00 microM and 5.2% at 0.100 microM. The lower limit of quantification and the limit of detection were 0.050 microM and 0.010 microM, respectively.