临床常规生化项目室内质控方法的设计

目的：探讨保证临床质量要求的常规生化项目室内质控规则的设计。方法收集2012年7—12月碱性磷酸酶(ALKP)、钾、钙的质控数据,以室内质控的累积变异系数(CV)作为项目的不精密度,应用卫生部临床检验中心室间质评结果计算偏倚,选择美国临床实验室改进修正法规88能力比对检验评价限为总允许误差,借助操作过程规范图设计符合本实验室切实可行的室内质控方法,从而提高误差检出概率( Ped),降低假失控概率( Pfr)。结果ALKP、钾每天测定质控品个数为2,选择1-3 s单质控规则即可满足Ped>90%,Pfr<5%;钙每天测定质控品个数为4,选择1-3 s/2-2 s/R-4 s/4-1 s/12-X作为质控规则,满足Ped>90%,Pfr<5%。结论不同生化分析项目应根据分析方法性能特征的不同,选择满足临床质量要求的控制方法。     

ELISA定性试验同一质控物不同批号试剂间连续质控法的建立与应用

目的 全自动酶免分析系统定性酶联免疫吸附法(ELISA)测定同一乙型肝炎表面抗原(HBsAg)质控物不同批号试剂间连续Levey-Jennings质控图法的建立与应用.方法 每批试剂均测定超低温保存可报告范围内不同浓度的5份HBsAg阳性标本的S/CO值,始终以原批号试剂测定S/C0值为Y,新批号试剂测定S/C0值为X1,得一直线回归方程Y=bX1 a;在换用新批号试剂时,得一新的直线回归方程(Y=bX2 a);依此类推.每个工作日均以浓度为2 ng/ml的HB sAg质控血清测定的S/CO值(X)通过该批号直线回归方程计算Y值连续作Levey-Jenning s质控图.结果 超低温保存不同浓度的5份HBsAg阳性标本的S/CO值在5批新试剂与原批号试剂间的直线回归方程分别为Y=3.2 38X1 0.48、Y=3.165X2 0.62、Y=2.966X3 0.85和Y=3.128X4 0.624、Y=3.25X5 0.54.结论 同一质控物在不同批号试剂间可以连续作Levey-Jennings质控图,该质控法有效、实用.     

应用固定范围法开展定性ELISA质量控制初探

目前,国内实验室大多采用定量试验的质控方法(如:Levey-Jennings质控图法﹑多规则的质控方法)开展ELISA定性试验的统计学室内质控,由于受到ELISA本身方法学特点的影响,这些方法在实际应用中会出现很多问题.近年也有学者介绍了美国CLIA'88最终规则规定的双重响应室内质控图法[1],用于检出假阳性反应和假阴性反应,但忽略了对测定精密度的监控,不利于检验质量的持续改进.本文应用固定范围法对定性ELISA统计质控方法进行了初步探讨,兼顾了两类方法的优点.     

肝炎综合征患儿肝功能、血脂及维生素E浓度的相关性分析

目的探讨肝炎综合征患儿血清胆红素、转氨酶、血脂及维生素E浓度的相关性。方法回顾性分析60例确诊为非胆道闭锁的肝炎综合征患儿的血清转氨酶、血脂及维生素E浓度的相关性。结果直接胆红素与总胆固醇呈正相关(r=0.314,P=0.015),与维生素E呈负相关(r=-0.435,P=0.001),丙氨酸氨基转移酶和谷氨酸氨基转移酶与维生素E呈负相关(r=-0.361,P=0.006;r=-0.421,P=0.001)。30例患儿补充维生素E后,TC、TG、TBIL及DBIL水平均显著降低(P<0.05或P<0.01)。结论补充维生素E可保护肝细胞膜,辅助改善婴儿肝炎综合征患儿的肝功能,进而促进胆汁排泄,降低胆红素和血脂水平。     

薄层扫描法测定固脾通鼻片中黄芪甲苷含量

目的测定固脾通鼻片中黄芪甲苷含量.方法 0.2%羧甲基纤维素钠作粘合剂的硅胶G薄层板;以氯仿-甲醇-水(40∶13∶1)溶液为展开剂;λs=510nm,λR=690nm.结果线性范围0.266～2.128μg(r=0.996),平均加样回收率为95.6%;RSD=2.64%(n=5).结论本法简便、准确、重现性好,可用于质量控制.     

不同检测系统胆红素测定结果的偏倚分析

目的:探讨不同检测系统间胆红素测定结果是否具有可比性。方法:以日立7170A生化分析仪,罗氏原装试剂及程序,c.f.a.s校准品和质控品组成的检测系统1作为目标检测系统,检测系统2和3为待评系统,在21个工作日内分别测定朗道质控物(水平2、水平3)各21次和新鲜血清样本45份。结果:经方差分析,朗道质控物和新鲜样本总胆红素(TBIL)、直接胆红素(DBIL)测定值各检测系统间的总体差异均有显著性(P<0.01),各系统间的相关系数大于0.975,临床可接受性能评价为检测系统2的TBIL结果部分超过允许范围,检测系统2的DBIL和检测系统3的TBIL、DBIL均超过允许范围。结论:3个检测系统测定TBIL、DBIL的结果之间存在不可比性。     

反向点杂交技术应用于β-地中海贫血基因研究

目的 应用反向点杂交技术(RDB)建立敏感．特异、简便的检测方法来同时检测17种不同的β-珠蛋白基因突变。方法 应用反向点杂交技术对734例来我院做产前检查的高危患者进行检测。结果 发生突变位点共68例,占9.26％。其中,654／N18例占2．45％,41～42／N31例占4．22％,22～28／N5例占0．68％,BE／N5例占0．41％,-28／N4例占0.54％,17／N4例占0．54％,14～15／N1例占0．14％,-29／41～421例占14％,71～72／N1例4占0．14％。结论 反向点杂交方法应用于进行β-地中海贫血的血液学筛查、基因诊断、携带者检出具有简便、快捷、准确的特点。     

用酶增强免疫分析法监测他克莫司血药浓度的质控评估

目的：评价酶增强免疫分析法（EMIT）监测患者全血中他克莫司浓度的质量,建立并改进他克莫司血药浓度监测的质量控制方法。方法：以标准质控为样本,进行预防性质量控制和室内质量控制研究。对2008年血药浓度监测中随行质控样本的测定值做回顾性研究并进行统计学分析,建立新的质控规则。结果：他克莫司低、中、高浓度（4.3、8.9、18.0 ng/ml）2009年质控样本的日内、日间RSD为1.8%-11.2%,平均回收率为90.5%-114.0%,符合《中华人民共和国药典》生物样品测定的要求。2008年质控样本的低、中、高浓度（3.8、7.5、15.0 ng/ml）的随行质控RSD分别为21.8%、14.2%和15.5%,适合本单位的质控规则为12S/13S/22S/41S/7tr。结论：EMIT法准确度和精密度良好,是一种较理想的他克莫司血药浓度测定方法,但影响因素较多,特别是温度对其影响较大,因此必须做好质量控制。     

反向点杂交技术应用于β-地中海贫血基因研究

目的 应用反向点杂交技术(RDB)建立敏感、特异、简便的检测方法来同时检测17种不同的β-珠蛋白基因突变。方法 应用反向点杂交技术对734例来我院做产前检查的高危患者进行检测，发生突变位点共68例，占92.6％。其中，654／N 18例占2.45％，41-42／N 31例占4．22％，22-28/N 5例占0.6％，BE／N 5例占0．41％，-28/N 4例占0.54％，17／N 4例占0.5％，14-15／N 1例占0．14％，-29／41-42 1例占14％，71-72／N 1例占0．14％。结论 反向点杂交方法应用于进行β-地中海贫血的血液学筛查、基因诊断、携带者检出具有简便、快捷、准确的特点。     

反相高效液相色谱法分离测定他达拉非的含量

目的建立分离测定他达拉非及其产品的反相高效液相色谱法。方法以Dionex Kromasil ODS1柱(250 mm×4．6 mm,5μm)为色谱柱,流动相为甲醇-乙腈-用三乙胺调节pH值至3．0的0．05 mol／L磷酸溶液(44:13:43),流速为1．5 mL／min,检测波长为290 nm,进样量20μL。结果他达拉非线性浓度范围为11．2-86．7μg／mL,r=0．999 9(n=6),平均回收率为100．20％,RSD为0．58％(n=6),重复进样RSD为0．38％(n=5),检出限为0．9 ng(S／N=3)。结论该法简便、快速、准确,适用于测定他达拉非及其产品的含量。     

4R危机管理理论在脓毒症患者胃肠功能障碍预防管理中的应用

目的　探讨4R危机管理理论在脓毒症患者胃肠功能障碍预防管理中的应用效果。方法　选择2020年12月至2021年12月广东省第二中医院收治的82例脓毒症患者为研究对象进行前瞻性研究,运用随机数字表法将其分为对照组和观察组,各41例。对照组男23例、女18例,年龄（76.17±9.27）岁,实施脓毒症常规护理干预;观察组男22例、女19例,年龄（77.22±9.03）岁,在对照组基础上实施基于4R危机管理理论的胃肠功能障碍预防管理。比较两组患者急性生理与慢性健康评分系统Ⅱ（APACHEⅡ）评分、胃肠功能损伤评分、胃肠功能障碍发生率以及护理满意度。计量资料行独立样本t检验,计数资料行χ²检验。结果　干预后,观察组APACHEⅡ评分、胃肠功能损伤评分［（19.41±3.15）分、（24.85±4.49）分］均低于对照组［（23.27±3.41）分、（29.07±5.12）分］,差异均有统计学意义（t=5.315、3.969,均P<0.001）。干预后,观察组患者胃肠功能障碍发生率为7.32%（3/41）,明显低于对照组24.39%（10/41）,差异有统计学意义（χ²=4.479,P=0.034）。干预后,观察组患者护理总满意度为97.6%（40/41）,高于对照组85.4%（35/41）,差异有统计学意义（χ²=3.905,P=0.048）。结论　运用4R危机管理理论可改善脓毒症患者病情和胃肠功能,降低胃肠功能障碍发生率,提高护理满意度。     

Dual agents loaded PLGA nanoparticles: Systematic study of particle size and drug entrapment efficiency

PLGA nanoparticles simultaneously loaded with vincristine sulfate (VCR) and quercetin (QC) were prepared via O/W emulsion solvent evaporation. Six independent processing parameters and PLGA characteristics were assessed systematically to enhance the incorporation of the dual agents with different properties (VCR and QC, hydrophilic and hydrophobic molecule, respectively) into PLGA nanoparticles and control particle size. Approaches investigated for the enhancement of drug entrapment efficiencies and the controlling of particle size included the influence of the molecular weight (MW) of PLGA and the lactide-to-glycolide (L:G) ratio of PLGA, PLGA concentration, PVA concentration, initial QC content, acetone-to-dichloromethane (A/D) volume ratio, aqueous phase pH and aqueous to organic phase (W/O) volume ratio. The nanoparticles produced by optimal formulation were submicron size (139.5+/-4.3 nm, n=3) with low polydispersity index (0.095+/-0.031, n=3). Nanoparticles observed by transmission electron microscopy (TEM) showed extremely spherical shape. The entrapment efficiencies determined by high performance liquid chromatography (HPLC) by ultracentrifuge method were 92.84+/-3.37% for VCR and 32.66+/-2.92% for QC (n=3). The drug loadings were 0.0037+/-0.0001% for VCR and 1.36+/-0.12% for QC (n=3).     

LC-MS/MS method for simultaneous determination of viramidine and ribavirin levels in monkey red blood cells

A high performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) method has been developed for the simultaneous determinations of total viramidine (viramidine, viramidine monophosphate, viramidine diphosphate, and viramidine triphosphate) and total ribavirin (ribavirin, ribavirin monophosphate, ribavirin diphosphate, and ribavirin triphosphate) in monkey red blood cells (RBC). The method involves the addition of internal standards and perchloric acid, conversion of viramidine or ribavirin phosphorylated metabolites to viramidine or ribavirin, purification with an aminopropyl (NH(2)) solid phase extraction (SPE) cartridge, and LC-MS/MS analysis. The MS/MS is selected to monitor m/z 245-->113, 250-->113, 244-->112, and 249-->112 for ribavirin, [(13)C]ribavirin, viramidine, and [(13)C]viramidine, respectively, using positive electrospray ionization. The calibration curves are linear over a concentration range of 100-10,000 ng/mL (0.412-41.2 microM) with a lower limit of quantification (LLOQ) of 100 ng/mL for both compounds. Mean inter-assay recoveries for ribavirin are 101%, 98.9%, and 96.0%, with coefficient of variance (%CV) values between 1.95 and 4.50% for 100, 1000, and 10,000 ng/mL quality control (QC) samples, respectively. Mean inter-assay recoveries for viramidine are 96.3%, 101%, and 102%, with coefficient of variation (%CV) values between 3.61 and 7.22%, for 100, 1000, and 10,000 ng/mL QC samples, respectively. Over-curve dilution QC at 400 microg/mL (1639 microM) for both viramidine and ribavirin are used to ensure the dilution accuracy (25 X dilutions) for monkey samples. The method has been used to simultaneously determine the total concentrations of ribavirin and viramidine in monkey RBC following 5, 15, and 36 weeks dosing of viramidine or ribavirin (60 mg/kg). The concentrations of total ribavirin following ribavirin dosing are 1242 microM at week 5, 1257 microM at week 15, and 1146 microM at week 36. The concentrations of total ribavirin following viramidine dosing are 634 microM at week 5, 716 microM at week 15, and 683 microM at week 36. Only small amounts of viramidine are detected in RBC following viramidine dosing, 7.80 microM at week 5, 6.63 microM at week 15, and 10.4 microM at week 36. The results suggest that ribavirin levels in RBC were at steady state at week 5 of ribavirin or viramidine dosing. At steady state, ribavirin levels in RBC are approximately 2x after ribavirin dosing than viramidine dosing. The relatively small percentage of viramidine in RBC suggests that viramidine either poorly penetrated into RBC or was extensively converted to ribavirin following entry into RBC.     

重氮法与钒酸盐氧化法测定总胆红素的方法学比较

目的分析重氮法与钒酸盐氧化法测定血清总胆红素结果的相关性,并对两法进行偏倚评估,建立仪器间的校正方法。方法依据美国国家临床实验室标准委员会（NCCLS）EP9．A文件方案,每天随机抽取8份血清标本,分别用重氮法与钒酸盐氧化法进行总胆红素测定,共测定5d。对检验结果进行离散点检查,计算线性方程和相关系数并进行偏倚估计。结果以重氮法（X）为参比方法,对钒酸盐氧化法（Y）进行评估,两者测定总胆红素的回归方程为Y=1．1588X-2．4616,相关系数R2=0．998。总胆红素浓度为34．2、171μmol／L、342μmol／L时,钒酸盐氧化法的相对偏倚分别为8．72％、14．42％、15．12％。结论两种方法测定血清总胆红素具有良好的相关性,其比例误差是0．1588,恒定误差是．2．4616μmol／L。其相对预期偏倚随总胆红素浓度增高而增高。     

Quantification of recombinant human parathyroid hormone (rhPTH(1-84)) in human plasma by immunoassay: commercial kit evaluation and validation to support pharmacokinetic studies

Immunoassays utilizing commercial kits designed for diagnostic use can be adapted and validated to meet Good Laboratory Practice (GLP) requirements to support pharmacokinetic (PK) studies. We illustrate in this paper a systematic approach for commercial kit evaluation and GLP-compliant method validation to establish selectivity, sensitivity, linearity, accuracy, precision and stability. Immunoassay kits for human parathyroid hormone (hPTH) quantification from three different vendors were assessed in a side-by-side comparison for their suitability for the PK analysis of recombinant humanPTH (rhPTH) in EDTA plasma. Two immunoradiometric (IRMA) assay kits and one immunoluminometric assay (ILMA) kit were evaluated. Since PTH is present as an endogenous component of human plasma, QC preparation in the biological matrix was handled differently than for a xenobiotic drug compound. The endogenous concentration of PTH was determined in plasma samples from 32 individual lots using the three kits. The lots with the lowest endogenous concentrations of PTH were selected, pooled to form the low QC and spiked with rhPTH to prepare the mid and high QCs. Four evaluation batches were run with each of the three commercial kits to evaluate reference standard linearity, and QC accuracy and precision. Selectivity against PTH peptide fragments PTH(7-84) and PTH(3-84) were assessed by cross-reactivity and accurate spike-recovery to the QC samples at two concentrations. One of the kits was chosen for full method validation because it had the lowest cross-reactivity against hPTH fragments (3-84) and (7-84), a wider dynamic range and the least total error. The accuracy and precision from six validation batches of the QCs were 相似文献   

Safety and pharmacology of paclitaxel in patients with impaired liver function: a population pharmacokinetic-pharmacodynamic study

AIMS: To assess quantitatively the safety and pharmacology of paclitaxel in patients with moderate to severe hepatic impairment. METHODS: Solid tumour patients were enrolled into five liver function cohorts as defined by liver transaminase and total bilirubin concentrations. Paclitaxel was administered as a 3-h intravenous infusion at doses ranging from 110 to 175 mg m(-2), depending on liver impairment. Covariate and semimechanistic pharmacokinetic-pharmacodynamic (PK-PD) population modelling was used to describe the impact of liver impairment on the pharmacology and safety of paclitaxel. RESULTS: Thirty-five patients were included in the study, and PK data were assessed for 59 treatment courses. Most patients had advanced breast cancer (n = 22). Objective responses to paclitaxel were seen in four patients (11%). Patients in higher categories of liver impairment had a significantly lower paclitaxel elimination capacity (R2 = -0.38, P = 0.05), and total bilirubin was a significant covariate to predict decreased elimination capacity with population modelling (P = 0.002). Total bilirubin was also a significant predictor of increased haematological toxicity within the integrated population PK-PD model (P < 10(-4)). Data simulations were used to calculate safe initial paclitaxel doses, which were lower than the administered doses for liver impairment cohorts III-V. CONCLUSIONS: Total bilirubin is a good predictor of paclitaxel elimination capacity and of individual susceptibility to paclitaxel-related myelosuppression in cancer patients with moderate to severe liver impairment. The proposed, adapted paclitaxel doses need validation in prospective trials.     

医疗机构临床试验质量控制标准实践研究

目的:建立机构内部的临床试验质量控制标准,规范临床试验质量控制工作.方法:根据质量控制实施阶段及侧重点,对质控进行分类;制定以项目启动质控细则及知情同意书、筛选入选表、研究者文件夹等单元模块质控细则,并依照已制定的质量控制标准实施质量控制.结果:制定以质控分类、质控标准细则、质控流程、质控问题判定为内容的临床试验质量控...     

改良鸟巢式护理干预在新生儿黄疸患儿中的应用效果

目的　探讨新生儿黄疸患儿采用改良鸟巢式护理的效果。方法　采用前瞻性研究,选取2020年1月至2021年12月在郑州大学附属儿童医院接受蓝光治疗的新生儿黄疸患儿80例,采用随机数字表法分为2组,各40例。观察组：男21例,女19例;胎龄30～41周;采用改良鸟巢式护理干预。对照组：男23例,女17例;胎龄31～41周;采用常规鸟巢式护理干预。统计两组患儿的基线资料,记录两组相关指标（日睡眠时间、日哭闹时间、住院时间、体温波动）,比较两组护理前、护理1 d、护理3 d及护理5 d时的胆红素［间接胆红素（IBIL）及总胆红素（TBIL）］水平,统计两组患儿不良反应情况,并评估两组家属对护理的满意度。统计学方法采用t检验、χ²检验。结果　观察组体温波动比对照组小［（0.35±0.10）℃比（0.83±0.12）℃］,日哭闹时间和住院时间［（45.37±4.09）min、（4.83±0.82）d］均比对照组［（56.73±5.82）min、（6.08±0.78）d］短,日睡眠时间比对照组长［（21.12±0.79）h比（17.26±2.93）h］,差异均有统计学意义（t=19.435、10.100、6.986、8.045,均P<0.001）;护理1 d、3 d、5 d时,两组血清IBIL、TBIL水平均依次较护理前降低,且观察组比对照组低（均P<0.05）;观察组患儿不良反应发生率比对照组低［5.00%（2/40）比20.00%（8/40）］,差异有统计学意义（χ²=4.114,P=0.043）;观察组家属护理满意度高于对照组［95.00%（38/40）比77.50%（31/40）］,差异有统计学意义（χ²=5.165,P=0.023）。结论　新生儿黄疸患儿采用改良鸟巢式护理干预可改善睡眠质量,减少哭闹时间及并发症,调节患儿的胆红素水平,将患儿家属的护理满意度提高。     

Preparation of clinical‐grade 89Zr‐panitumumab as a positron emission tomography biomarker for evaluating epidermal growth factor receptor‐targeted therapy

Panitumumab is a fully human monoclonal antibody approved for the treatment of epidermal growth factor receptor (EGFR) positive colorectal cancer. Recently, panitumumab has been radiolabeled with ⁸⁹Zr and evaluated for its potential to be used as immuno‐positron emission tomography (PET) probe for EGFR positive cancers. Interesting preclinical results published by several groups of researchers have prompted us to develop a robust procedure for producing clinical‐grade ⁸⁹Zr‐panitumumab as an immuno‐PET probe to evaluate EGFR‐targeted therapy. In this process, clinical‐grade panitumumab is bio‐conjugated with desferrioxamine chelate and subsequently radiolabeled with ⁸⁹Zr resulting in high radiochemical yield (>70%, n = 3) and purity (>98%, n = 3). All quality control (QC) tests were performed according to United States Pharmacopeia specifications. QC tests showed that ⁸⁹Zr‐panitumumab met all specifications for human injection. Herein, we describe a step‐by‐step method for the facile synthesis and QC tests of ⁸⁹Zr‐panitumumab for medical use. The entire process of bioconjugation, radiolabeling, and all QC tests will take about 5 h. Because the synthesis is fully manual, two rapid, in‐process QC tests have been introduced to make the procedure robust and error free. Copyright © 2013 John Wiley & Sons, Ltd.     

茵栀黄口服液治疗新生儿病理性黄疸疗效观察

目的 观察茵栀黄口服液治疗新生儿病理性黄疸的临床疗效。方法 将新生儿病理性黄疸81例随机分为对照组40例和治疗组41例,对照组采用西医常规治疗,治疗组在对照组的基础上给予茵栀黄口服液治疗,观察两组患儿临床疗效、黄疸消退时间及血清胆红素变化情况。结果 两组患儿治疗总有效率差异无统计学意义（P〉0.05）,但显效率比较,治疗组优于对照组（P〈0.01）,治疗组黄疸消退时间快于对照组（P〈0.05）,且在治疗后第3、6、8d治疗组患儿血清胆红素水平低于对照组（P〈0.05）。结论 茵栀黄口服液对于治疗新生儿高胆红素血症有较好疗效。