目的 比较高效液相色谱-串联质谱法(HPLC-MS/MS)和化学发光微粒子免疫检测技术(CMIA)测定人全血他克莫司(FK506)浓度结果的相关性,以及2种方法在他克莫司浓度监测工作中的应用.方法 收集253例心脏移植患者(n=581)术后服用他克莫司的全血样本,采用HPLC-MS/MS法测定人全血他克莫司血药浓度,通过专属性、标准曲线与定量下限、精密度、准确度、基质效应和提取回收率以及稳定性试验进行方法学验证;并与CMIA法的测定结果进行比较,观察、比较2种检测手段的测定结果及相关性.结果 用HPLC-MS/MS测定人全血他克莫司浓度,线性范围2～30 ng·mL-1,定量下限为2ng· mL-1;日内及日间精密度(RSD％)均小于15％.按照Bland-Altman法计算,95.52％ LoA=(0.22,5.62),95.52％的差值都位于一致限内,两种方法测定值一致性良好.回归方程为FK506 HPLC-MS/MS=0.70×FK506CMIA-0.17,Pearson相关系数为0.902 4(P ＜0.05),说明两种方法具有良好的相关性.结论 HPLC-MS/MS法测定他克莫司血药浓度定量特异性强,灵敏度高.与CMIA法相比,HPLC-MS/MS测定值低,对他克莫司母药有高度的选择性.

Objective To investigate the effect of age and gender on blood pressure (BP) rhythm of patients with essential hypertension based on 24-hour ambulatory blood pressure.Methods 312 patients with essential hypertension were analyzed by 24-hour ambulatory blood pressure monitoring (ABPM).They were divided into four groups according to age:the ＜ 40-year-old group,40-49-year-old group,50-59-year-old group and the ≥ 60--year-old group.The influence of age and gender on the characteristics of blood pressure rhythm and morning hypertension was studied.Results The incidence of dippers,non-dippers,reverse dippers and ultra-dippers was 45.19％,41.35％,7.37％ and 6.09％,respectively.The proportion of dippers in the youngest group (65％) was the highest while reverse dippers were prevalent(18.92％) in the elderly group.Gender had no significant effect on blood pressure type.Only reverse dippers were more common in the female group than in the male group(P ＜ 0.01).Cases of morning hypertension accounted for 96.5％.Age had no effect on the incidence of morning hypertension,but there was significant difference between the male group (98.6％) and the female group (92.4％) in this regard.Conclusions Elderly patients with abnormal circadian rhythm outnumber younger patients.Male patients are more prone to morning hypertension.More attention should be paid to abnormal circadian rhythm,especially to morning hypertension,in order to individualize antihypertensive treatments.

Purpose The goal of this study was to develop a population pharmacokinetic (PK) and PK/pharmacodynamics (PD) model for ibutilide, to evaluate the time course of its effect on QT interval in Chinese. Methods The population PK and PK/PD model were developed using data from 40 Chinese healthy volunteers using nonlinear mixed-effects modeling, and the final population PK/PD model was applied on 100 patients with atrial fibrillation (AF) and/or atrial flutter (AFL). Findings The PK parameters of ibutilide were best described by a 3-compartment model with first-order elimination. No statistically significant covariate was found for each PK model parameter. Individualized QT interval correction, by heart rate, was performed by a power model, and the circadian rhythm of QT intervals was described by 2 mixed-effect cosine functions. The QT interval data of ibutilide was well characterized by a sigmoid Emax model ( E ( C ) = E m a x γ × C γ / ( EC 50 γ + C γ ) ) with an effect compartment. The final PK/PD model was used to estimate individual parameters of patient data and found good predictions compared with healthy volunteers; AF and/or AFL patients had lower Emax and higher EC50. Implications A population PK and PK/PD model for ibutilide in healthy volunteers was developed and could well capture ibutilide’s PK/PD characteristics. The final PK/PD model was applied on patients with AF and/or AFL successfully.

Objective To establish the population pharmacokinetic and pharmacokinetic models of ibutilide in healthy subjects,and to study the dose-exposure-drug effect relationship and variation of ibutilide in healthy subjects and to provide the basis for clinical practice of ibutilide.Methods A total of 40 healthy subjects were enrolled in this single-center,randomized,dose-escalation trial.13 plasma concentrations of ibutilide and 29 QT interval observations were collected from each subject.NONMEM 7.2 was used to establish and evaluate the pharmacokinetic and pharmacodynamic models.Results The pharmacokinetic model of ibutilide was best described by the three-compartment model with linear elimination,and the pharmacodynamic model was rationally characterized by the sigmoid Emax model.Goodness-of-fit (GOF) was validated by visual predictive check(VPC) and Bootstrap,which indicated that the population pharmacokinetic and pharmacodynamic models of ibutilide in healthy subjects were stable and reliable.Parameters with high confidence were obtained,and the individual variation of population pharmacodynamic parameters the exponent of the sigmoid equation (γ) and the maximum response of QT interval (Emax) were 133.50％ and 64.10％,respectively.Conclusion The population pharmacokinetic and pharmacodynamic models of ibutilide in healthy volunteers were established for the first time and the presence of ibutilide-sensitive and poor-response population were observed,which suggested the necessity of individualized administration for ibutilide.

Objective:To evaluate the linearity of Coulter automated hematology analyzer LH 780. Methods:The linearity control used to be samples, the linear range of WBC, RBC, Hgb, Plt were evaluated with Coulter original reagents, calibrator and control. Results:The test results of the four parameters showed good linear relationship. The linear range of WBC, RBC, Hgb, Plt were (0.03~363.5)×109 /L, (0.00~7.92)×1012/L, 0.3~236.0 g/L and (0.3~2511.0)×109/L, respectively. Conclusion:For Coulter hematology analyzer LH 780, there is a wide linearity that can meet the demands of clinical laboratories.

目的 探讨冠状动脉造影与支架置入术对腺苷二磷酸(ADP)诱导的血小板聚集率的影响.方法 前瞻性纳入2012年5月1日至2013年4月30日中国医学科学院阜外医院冠心病患者343例,术前至少7d连续服用阿司匹林100 mg,每日1次;氯吡格雷75 mg,每日1次.根据支架置入情况分为单纯冠状动脉造影组(造影组,173例)和支架置入组(170例).患者在接受冠状动脉介入操作之前及之后24 h内,分别采集空腹血样本,用光学比浊法测定血小板聚集率,比较两组患者术前、术后血小板聚集率的变化,支架置入组有66例患者自愿参加血小板聚集率复测亚组分析.结果 两组患者术前、术后血小板聚集率比较,差异无统计学意义(P＞0.05);造影组患者术前、术后血小板聚集率比较,差异无统计学意义(P=0.062),而支架置入组患者术后血小板聚集率显著高于术前[(55.59±10.47)％比(52.47±11.97)％,P＜0.001],差异有统计学意义;支架置入组患者术后血小板聚集率增加大于造影组[(3.12±8.31)％比(1.06±7.40)％,P=0.010],差异有统计学意义.随着患者支架置入数量的增加,术后血小板聚集率呈递增趋势,但差异无统计学意义(P＞0.05).支架置入组血小板聚集率复测亚组患者(66例)术后30 d血小板聚集率显著低于术后24 h[(54.71±11.64)％比(56.68±10.21)％,P=0.019],且与术前基线值比较,差异无统计学意义[(54.71±11.64)％比(54.26±12.23)％,P=0.901].结论 冠状动脉支架置入操作可导致术后血小板活性升高,该作用可在术后30 d内消失,而单纯冠状动脉造影则无此影响.

A liquid chromatography-tandem mass spectrometry (LC-MS) method to quantify tolvaptan and its two main metabolites and applied to human study was first developed and validated as a measure of compliance in clinical research. Because of the structure similarity of tolvaptan and its multiple metabolites, the method was optimized to obtain a chromatographic and MS separation of the endogenous interference and isotope ions as well as high analysis throughput. Tolvaptan, its two main metabolites and the internal standard were extracted from human serum (0.1mL) using solid-phase extraction, separated on a Waters nova-pak C18 column (150×3.9mm, 5μm) using isocratic elution with a mobile phase composed of acetonitrile, water and formic acid (65:35:0.25, v/v/v). The total run-time was shortened to 3.5min. The mass transition ranges under positive electrospray ionisation that were monitored for quantitation included m/z 449-252 for tolvaptan, m/z 479-252 for metabolite DM-4103, m/z 481-252 for metabolite DM-4107 and m/z 463-266 for the internal standard (IS). The limit of quantification in plasma for all three analytes was 1ng/mL. The method was validated over a linear range from 1 to 500ng/mL for all three analytes with acceptable inter- and intra-assay precision and accuracy. The stability of the analytes was determined to be suitable for routine laboratory practices. The method was successfully applied to samples taken from research volunteers who ingested a 15mg tolvaptan tablet.

目的 对ACL TOP 500全自动凝血分析仪进行检测性能评价,包括精密度、准确度和线性范围,并验证参考区间.方法 使用原装配套试剂、校准品和质控品,评价凝血酶原时间(PT)、活化的部分凝血活酶时间(APTr)和纤维蛋白原(FIB)的精密度和准确度,至少20次质控品的检测结果用于评价精密度;20份样本的检测结果与另一台同型号仪器比对用于评价准确度;采用5份样本评价FIB的线性范围;20份健康个体样本用于验证上述3个项目及国际标准化比值(INR)的参考区间.结果 所有项目两水平质控品检测结果的变异系数均＜3.6％;全部样本各项目与同型号仪器的比对结果的差异在总误差范围内;FIB的线性范围为0.52 g/L ～ 7.74 g/L;拟验证的参考区间可接受.结论 ACLTOP 500全自动凝血分析仪的检测性能良好,能满足临床实验室的要求.

本文尝试在质量管理体系基础上,嵌入风险管理理念,探讨药物临床试验机构的风险管理模式.根据临床试验相关法规与风险管理通用原则,通过风险识别、风险评估和风险处置,将32项风险因素纳入至临床试验项目风险管理表中,初步建立药物临床试验机构风险管理模式,以此改进药物临床试验项目的管理质量,有效降低风险的发生率.

BACKGROUND:As an acute phase protein, α1-antitrypsin (AAT) has been extensively studied in acute coronary syndrome, but it is unclear whether a relationship exists between AAT and stable angina pectoris (SAP). The purpose of the present study was to investigate the association between AAT plasma levels and SAP.METHODS:Overall, 103 SAP patients diagnosed by coronary angiography and clinical manifestations and 118 control subjects matched for age and gender were enrolled in this case-control study. Plasma levels of AAT, high-sensitivity C-reactive protein (hsCRP), lipid profiles and other clinical parameters were assayed for all participants. The severity of coronary lesions was evaluated based on the Gensini score (GS) assessed by coronary angiography.RESULTS:Positively correlated with the GS (r = 0.564, P < 0.001), the plasma AAT level in the SAP group was significantly higher than that in the control group (142.08 ± 19.61 mg/dl vs. 125.50 ± 19.67 mg/dl, P < 0.001). The plasma AAT level was an independent predictor for both SAP (odds ratio [OR] = 1.037, 95% confidence interval [CI]: 1.020-1.054, P < 0.001) and a high GS (OR = 1.087, 95% CI: 1.051-1.124, P < 0.001) in a multivariate logistic regression model. In the receiver operating characteristic curve analysis, plasma AAT level was found to have a larger area under the curve (AUC) for predicting a high GS (AUC = 0.858, 95% CI: 0.788-0.929, P < 0.001) than that of hsCRP (AUC = 0.665, 95% CI: 0.557-0.773, P = 0.006; Z = 2.9363, P < 0.001), with an optimal cut-off value of 137.85 mg/dl (sensitivity: 94.3%, specificity: 68.2%).CONCLUSIONS:Plasma AAT levels correlate with both the presence and severity of coronary stenosis in patients with SAP, suggesting that it could be a potential predictive marker of severe stenosis in SAP patients.

Objective: To investigate the diagnostic value of high-density lipoprotein (HDL) associated serum amyloid A (SAA) to apolipoprotein A-I (apoA-I) ratio (SAA/apoA-I) in patients with coronary artery disease (CAD). <br> Methods: A total of 152 patients who received coronary angiography in our hospital were studied. According to the degree of coronary stenosis, the patients were divided into 2 groups. CAD group,n=77 and Control group,n=75. HDL was isolated by density gradient ultracentrifugation and the levels of SAA and apoA-I both in HDL and plasma were examined by ELISA. <br> Results: Compared with Control group, CAD group had increased ratio of HDL-SAA/HDL-apoA-I,P<0.001. Multivariate logistic regression analysis indicated that the ratio of HDL-SAA/HDL-apoA-I was independently related to the risk of CAD (OR=9.521, 95% CI 3.345-27.100,P<0.001). Receiver operating characteristic curve (ROC) analysis presented that the ratio of HDL-SAA/HDL-apoA-I had the medium value for CAD diagnosis (AUC=0.712, 95% CI 0.632-0.793,P<0.001), the best cut-off value was at 3.27. The plasma ratio of SAA/apoA-I was positively related to the ratio of HDL-SAA/HDL-apoA-I (r=0.857,P<0.001), the ratio of SAA/apoA-I had the similar value for CAD diagnosis (AUC=0.676, 95% CI 0.591-0.761,P<0.001), the best cut-off value was at 8.3. <br> Conclusion: The ratio of SAA/apoA-I in HDL is positively related to the risk of CAD, it might be become a potential diagnostic marker in patients with CAD.

The purpose of this paper is to study a management model for clinical trial institute,which is based on the quality management systems(QMS) proposed in ISO 9000 international standard.The QMS in Fuwai Clinical Trial Institute was established using process approach in accordance with clinical trial legal or regulation,standard operating procedure(SOP) and our work requirements.The main activities include defining management responsibility,implementing processes,reviewing project management,finding problem,and taking action to continually improve.The QMS in our institute has efficiently improved clinical trial management and met drug registration requirements for the sponsor or Contract Research Organization(CRO) through Plan-Do-Check-Act methodology.

目的 通过对Coulter LH780血细胞分析仪自动和手动吸样模式检测结果比对,评价2种模式检测结果是否具有可比性.方法 采集20份抗凝(乙二胺四乙酸二钾,EDTA-K2)全血.每份样本分别用自动和手动吸样模式各检测2次,以自动吸样模式为参比模式,计算各项指标的相对差异,以国家卫生行业标准(WS/T406-2012)和t检验方法对检测结果进行分析.结果 红细胞计数(RBC)、白细胞计数(WBC)、血红蛋白浓度(HGB)项目检测结果为100％可接受,PLT项目检测结果90％可被接受,但差异无统计学意义(P＞0.05).结论 在严格执行仪器使用和保养标准操作规程和良好室内质控的基础上,血细胞分析仪自动和手动吸样模式的检测结果具有可比性.

目的:比较研究BCRP基因多态性对瑞舒伐他汀单剂量和多剂量连续给药后人体药动学的影响。方法:筛选24例健康受试者,按BCRP 421 C>A基因型分组:421CC野生组（n=15）和421CA+AA突变组（n=9）。受试者每日口服规定剂量的瑞舒伐他汀,连续7 d;每日按规定时间点采集受试者血样和尿样;采用液相色谱-串联质谱联用法（HPLC-MS/MS）测定血浆和尿样中的瑞舒伐他汀浓度。结果:单剂量给药后,瑞舒伐他汀在BCRP 421CA+AA突变组的AUC0～t,C max和尿药排泄百分数Percent e xcretion显著高于421CC野生型组（P=0.030,0.015和0.041）,半衰期t1/2和达峰时间T max在两组人群中无差异（P>0.05）。与单剂量结果不同,多剂量连续给药达稳态后,稳态AUC ss,C max、总清除率CL total s s/F和肾清除率CL R s s在野生型组和突变组之间均无显著性差异（P>0.05）,但半衰期t1/2在野生组显著延长[（14.1±3.1）vs（11.8±2.2）h,P=0.035],且野生组的蓄积比Rac也高于突变组（P=0.064）。结论:BCRP 421C>A基因多态性是影响瑞舒伐他汀人体药动学改变的重要因素。

目的 以总蛋白(TP)为例,考察Roche Cobas C501全自动生化分析仪的精密度、准确度和可报告范围,以验证其检测性能.方法 参考美国临床和实验室标准化协会(CLSI) EP5-A2、EP6-A等文件,使用C501的原装配套试剂及校准品,检测总蛋白(TP)的精密度、准确度和可报告范围,用EP Evaluator软件统计分析.结果 TP两水平精密度检测结果的批内标准差(S)、总的S均不超过验证值,精密度可以接受;在对照系统与实验系统之间,20个患者血清样本的检测结果的误差指数(EI)均在±1.00之间,准确度可以接受;覆盖可报告范围的5个水平样本中的4个结果呈线性,线性可接受.结论 Cobas C501分析仪TP项目的精密度、准确度、可报告范围达到了厂家声明的检测性能,此分析仪的检测性能得到了验证.

A liquid chromatography-tandem mass spectrometry method to quantify carvedilol enantiomers in human plasma was developed and validated as a measure of compliance in clinical research. Carvedilol enantiomers were extracted from human serum (0.5 mL) via liquid-liquid extraction with methyl tert-butyl ether (2.5 mL). Carvedilol-related compound C served as the internal standard. The analyte and internal standard were separated on a Sino-Chiral AD column (150 × 4.6mm, 5 μm, amylose tris-3,5-dimethylphenylcarbamate coated on silica-gel) using isocratic elution with mobile phases of methanol, water and diethylamine (94:6:0.01, v/v). The total run-time was 10.5 min. Carvedilol enantiomers were quantified using a triple quadrupole mass spectrometer operated in multiple-reaction-monitoring mode using positive electrospray ionisation. The mass transitions monitored for quantitation were carvedilol (m/z 407→222) and carvedilol-related compound C (m/z 497→222). The limits of quantification for the S- and R-carvedilol enantiomers in plasma were both 0.08 ng/mL. The method was validated in the linear range of 0.08-50 ng/mL with acceptable inter- and intra-assay precision and accuracy and stability suitable for routine laboratory practice. The method was successfully applied to samples taken from research volunteers treated with carvedilol sustained-release tablet 18 mg. Cmax and AUClast were 9.1 ± 5.1 ng/mL and 59.4 ± 39.6 ng h/mL for R-carvedilol, 4.0 ± 2.3 ng/mL and 24.7 ±15.0 ng h/mL for S-carvedilol, respectively. tmax and t1/2 were 4.6 ± 1.9h and 9.6 ± 4.5h for R-carvedilol, and 4.7 ± 1.0 h and 10.7 ± 5.7 h, respectively.

目的：研究表明，冠心病患者血浆高密度脂蛋白（HDL）组成的改变可能与冠心病风险相关。本研究拟探讨HDL组分中载脂蛋白A1（apoA1）和血浆淀粉样蛋白A（SAA）与冠心病风险的关系，及二者在冠心病诊断中的价值。

目的:探讨脂滴包被蛋白(perilipin,PLIN)基因多态性与原发性高血压患者美托洛尔治疗后甘油三酯升高的相关性。方法:入选患者97例,服用美托洛尔缓释片100 mg·d-1,持续8周;服药前后分别测量血糖、甘油三酯(TG)、总胆固醇(TC)、高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)水平;直接测序法分析PLIN rs2304796 C>T多态性;使用SPSS 15.0统计软件进行数据分析。结果:服用美托洛尔后,患者血浆TG较服药前升高[(2.05±2.42)vs(2.38±2.71)mmol·L-1,P=0.016)]。血糖、TC、HDL-C、LDL-C服药前后无明显变化。PLIN rs2304796 C>T多态性与美托洛尔服药后TG升高相关,其中,CC型携带者升高(0.23±1.02)mmol·L-1,CT型升高(0.26±1.07)mmol·L-1,TT型升高(1.40±3.21)mmol·L-1,P=0.047。结论:perilipin蛋白rs2304796 C>T多态性与美托洛尔服用后TG的升高相关,TT基因型携带者TG水平升高明显。