应用有限采样法估算头孢克洛在健康志愿者体内暴露程度的药动学参数

目的:应用有限采样方法(LSS)和图形化评价方法,建立1至4个时点血药浓度和时间时点的回归方程,估算头孢克洛体内暴露程度的药动学参数AUC0-240min和Cmax,为个体化用药提供依据及方法。方法:健康受试者口服用药头孢克洛250mg,应用LSS法对240min内血药浓度数据进行建模,并用Jackknife法验证模型,以绝对预测误差(APE)、误差均方及图形检测评价建模结果。结果:2至4个时点的LSS法可以准确地预测头孢克洛在体内暴露程度。Jackknife法验证结果显示,AUC0-240min用一个时点,48例数据中有23例(47.9%)的绝对误差超过15%,采用2、3、4个时点建模预测的结果较好,48例中分别有8(16.7%)、4(8.3%)和0例预测结果的APE超过15%。Cmax用一个时点,48例数据中有24例(50.0%)的绝对误差超过15%。采用2、3、4个时点建模预测结果也较好,48例中分别有8(16.7%)、7(14.6%)和7(14.6%)例预测结果的APE超过15%。结论:应用有限采样法,采用2至4个血药头孢克洛浓度即可较准确地预测AUC0-240min和Cmax,从而方便地指导个体用药。     

HPLC法测定丹参颗粒中丹酚酸B的含量

目的建立丹参养心颗粒中丹酚酸B含量的测定方法。方法采用HPLC法测定丹酚酸B的含量,色谱柱：OSD C18反相色谱柱（5μm,4.6×250 mm）;流动相：乙腈-2.5%甲酸（20：80）;流速：1.0 ml/min;检测波长：286 nm;柱温：35℃。结果平均回收率为99.55%,RSD%=0.42%。结论方法简便可行,重现性好,可很好地控制丹参养心颗粒的内在质量。     

毛细管电泳法测定丹参中丹酚酸B的含量

丹参(Salvia miltiorrhiza Bge．)为常用中药，具有去淤止痛，活血通经之功效。丹参的多种水溶性酚酸中以丹酚酸B的抗脂质过氧化和清除自由基作用最强，且在药材中含量最高…。有报道采用薄层扫描法和高效液相色谱法测定丹参中多种水溶性酚酸类成分，而毛细管电泳方法作者未见文     

从丹参中提取高纯度的丹参酚酸B

目的建立提取分离高纯度丹参酚酸B的方法。方法以HPLC-UV为检测手段,通过对提取温度、时间、用水量以及萃取次数等的优化,比较几种大孔吸附树脂对丹参酚酸B的吸附及洗脱性能,筛选制备高纯度丹参酚酸B的工艺条件。结果确定最佳工艺为加10倍量水、85℃恒温水浴提取1.5 h、提取2次,上清液用乙酸乙酯萃取4次,丹参酚酸B提取率为87.1%,纯度为95.5%。结论新建方法稳定高效,能较好地提取纯化丹参酚酸B。     

HPLC法测定丹参冲剂中丹酚酸B的含量

目的：采用HPLC法测定丹参冲剂中丹酚酸B的含量。方法：采用DiamonsilC18柱（5μ,200×4.6mm）;以甲醇-乙腈-甲酸-水（30：10：1：59）为流动相;流速：1.0ml·min-1;检测波长：286nm。结果：丹酚酸B在12.08～120.80μg·ml-1浓度范围内线性关系良好,平均加样回收率为99.42%（n=9）,RSD=0.53%。结论：建立的方法可准确测定丹参冲剂中丹酚酸B的含量。     

不同提取方法对丹参中丹酚酸B含量测定的影响

目的研究不同提取方法对丹参中丹酚酸B含量测定的影响。方法采用高效液相色谱法比较了煎煮、加热回流、索氏提取和超声处理4种不同提取方法中丹酚酸B的含量。结果超声方法提取的丹酚酸B含量比加热回流等方法高,差异明显。结论在提取丹参中丹酚酸B时,超声提取方法优于加热回流等方法。     

丹参在炮制大生产过程中丹酚酸B的损失情况研究

目的考察丹参药材炮制大生产过程中指标性成分丹酚酸B的含量变化及损耗情况。方法进行10批次丹参饮片炮制生产,应用HPLC法,在生产过程各主要工艺点进行跟踪取样,测定丹酚酸B的含量并计算损耗;同时在其中一个批次药材炮制大生产过程中,检测每个工艺环节对丹酚酸B的含量的影响。结果原药材、水洗药材、润透药材和切片4个取样点丹酚酸B的平均含量分别为6.83%,4.72%,4.46%和3.55%,相对原药材的损失情况分别为30.84%,34.59%和47.99%。水洗工艺直接导致丹酚酸B损失22.11%,最终损失高达47.48%。结论严格控制(优化)水洗工艺,可有效保留丹酚酸B的含量,提高丹参的炮制效率。     

改进丹参饮片的炮制方法提高丹酚酸B含量

目的研究中药材气相置换软化的机理,进行气相置换软化。方法采用气相置换润药机软化丹参与普通方法进行比较。结果采用气相置换润药设备软化丹参具有药材含水量率低,软化效果好,丹酚酸B含量高的特点。结论气相置换软化法使药材可在低含水量的情况下,快速、均匀软化,且可提高丹酚酸B的含量。     

基于脑微血管内皮细胞保护的注射用丹参多酚酸质量生物标志物研究

目的 基于血管内皮保护的关键药效环节,发现能够反映注射用丹参多酚酸（SAFI）的质量生物标志物（Qbiomarkers）,为完善其质量控制评价提供参考。方法 90℃加热处理SAFI 4、18、60 h,得到不同化学质量的SAFI（SAFI4、SAFI18、SAFI60）,高效液相色谱（HPLC）法分析紫草酸、丹酚酸B、丹酚酸Y、丹参素钠、原儿茶醛、丹酚酸D、迷迭香酸变化趋势。体外培养小鼠脑微血管内皮细胞（bEnd.3）,建立氧糖剥夺/复氧（OGD/R）模型,给予SAFI、SAFI4、SAFI18、SAFI60处理18 h,CCK-8法检测细胞相对存活率,试剂盒法检测上清液乳酸脱氢酶（LDH）、NO水平,划痕实验检测细胞迁移率;通过DIA蛋白质组学分析不同化学质量的SAFI给药后产生的差异蛋白,结合药效学及偏最小二乘回归（PLSR）分析筛选SAFI的Q-biomarkers。结果 与SAFI相比,SAFI4、SAFI18、SAFI60样品中的紫草酸、丹酚酸B和丹酚酸Y含量下降,丹参素钠含量上升,原儿茶醛、丹酚酸D、迷迭香酸变化不明显。药效学结果表明,与模型组相比,SAFI、SAFI4及SAFI18组细胞相对存活率显著上升、LDH泄漏量显著下降、NO水平显著下降、细胞迁移率显著升高（P＜0.05、0.01、0.001） ,对细胞具有显著保护作用;而SAFI60组无明显保护作用。蛋白质组学结果表明,18个差异蛋白的含量随药效学和成分的变化而发生一致性上升或下降,与药效进行相关性分析发现Kdr、Cxcl12的变量重要性投影值（VIP）皆大于1,尤其Kdr ［血管内皮生长因子受体2 （VEGFR2）］与血管内皮功能高度相关,可能是潜在的SAFI的Q-biomarkers。结论 通过不同化学质量的SAFI的内皮保护药效及差异蛋白关联分析发现,VEGFR2可以作为SAFI的Q-biomarker。     

丹参酚酸盐在健康志愿者的药代动力学和药效学研究

目的:研究丹参酚酸盐在健康人体的药代动力学,以及血药浓度与血小板粘附性和聚集率的关系。方法:9名健康志愿者根据交叉设计,静脉滴注丹参酚酸盐200、300、400 mg,用药后不同时间采血样,用高效液相色谱法测定丹参酚酸盐血药浓度;玻球法测定血小板粘附率;比浊法测定血小板聚集率。结果:丹参酚酸盐药动学代谢呈二室模型,消除半衰期约1 h,表观分布容积较大,提示为全身分布,峰浓度约为14 mg·L-1。血小板粘附率在用药后呈波动性下降,5 h(即用药后4 h)药效渐渐消失,可见一个药效滞后环,在停药后3 h、血药浓度在1.91mg·L-1时药效达峰值。血小板粘附率在静滴药物期间与药浓相关,药效峰值与药浓峰值吻合,也呈现药效滞后环。结论:丹参酚酸盐在健康人体消除快,对血小板聚集率和粘附率均有降低作用,前者达峰时间在滴注给药末,后者在停滴后3 h,均呈现较好的滞后效应。     

HPLC测定冠心丹参滴丸中丹参酮Ⅱ_A的含量

目的建立冠心丹参滴丸中丹参酮ⅡA的含量测定方法。方法运用高效液相色谱法测定,色谱柱为ZORBAXXDB-C18(4.6mm×150mm,5μm),流动相:甲醇-水(80∶20),流速1ml.min-1,检测波长270nm,柱温:室温。结果丹参酮ⅡA在0.0458~0.1026μg范围内峰面积与测定量呈良好的线性关系,回归方程为Y=6.7426×103X-3.5967×102(r=0.9999),平均加样回收率为97.75%,RSD为1.81%(n=5)。结论本法灵敏,快速,准确,可用于测定冠心丹参滴丸中丹参酮ⅡA的含量。     

Cigarette smoking substantially alters plasma microRNA profiles in healthy subjects

Circulating microRNAs (miRNAs) are receiving attention as potential biomarkers of various diseases, including cancers, chronic obstructive pulmonary disease, and cardiovascular disease. However, it is unknown whether the levels of circulating miRNAs in a healthy subject might vary with external factors in daily life. In this study, we investigated whether cigarette smoking, a habit that has spread throughout the world and is a risk factor for various diseases, affects plasma miRNA profiles. We determined the profiles of 11 smokers and 7 non-smokers by TaqMan MicroRNA array analysis. A larger number of miRNAs were detected in smokers than in non-smokers, and the plasma levels of two-thirds of the detected miRNAs (43 miRNAs) were significantly higher in smokers than in non-smokers. A principal component analysis of the plasma miRNA profiles clearly separated smokers and non-smokers. Twenty-four of the miRNAs were previously reported to be potential biomarkers of disease, suggesting the possibility that smoking status might interfere with the diagnosis of disease. Interestingly, we found that quitting smoking altered the plasma miRNA profiles to resemble those of non-smokers. These results suggested that the differences in the plasma miRNA profiles between smokers and non-smokers could be attributed to cigarette smoking. In addition, we found that an acute exposure of ex-smokers to cigarette smoke (smoking one cigarette) did not cause a dramatic change in the plasma miRNA profile. In conclusion, we found that repeated cigarette smoking substantially alters the plasma miRNA profile, interfering with the diagnosis of disease or signaling potential smoking-related diseases.     

Biomarkers for the effects of selective serotonin reuptake inhibitors (SSRIs) in healthy subjects

AIMS: Studies of novel centrally acting drugs in healthy volunteers are traditionally concerned with kinetics and tolerability, but useful information may also be obtained from biomarkers of clinical endpoints. This paper provides a systematic overview of CNS-tests used with SSRIs in healthy subjects. A useful biomarker should meet the following requirements: a consistent response across studies and drugs; a clear response of the biomarker to a therapeutic dose; a dose-response relationship; a plausible relationship between biomarker, pharmacology and pathogenesis. METHODS: These criteria were applied to all individual tests found in studies of selective serotonin reuptake inhibitors (SSRIs), performed in healthy subjects since 1966, identified with a systematic MedLine search. Separate databases were created to evaluate the effects of single or multiple dose SSRI-studies, and for amitriptyline whenever the original report included this antidepressant as a positive control. Doses of the antidepressant were divided into high- and low-dose ranges, relative to a medium range of therapeutic doses. For each test, the drug effects were scored as statistically significant impairment/decrease (-), improvement/increase (+) or no change (=) relative to placebo. RESULTS: 56 single dose studies and 22 multiple dose studies were identified, investigating the effects of 13 different SSRIs on 171 variants of neuropsychological tests, which could be clustered into seven neuropsychological domains. Low single doses of SSRIs generally stimulated tests of attention and memory. High doses tended to impair visual/auditory and visuomotor systems and subjective performance, while showing an acceleration in motor function. The most pronounced effects were observed using tests that measure flicker discrimination (improvement at low doses: 75%, medium doses: 40%, high doses: 43% of studies); REM sleep (inconsistent decrease after medium doses, decrease in 83% of studies after high doses); and EEG recordings, predominantly in alpha (decrease in 60% and 43% of studies after low and medium doses, respectively) and in theta activity (increase in 43% and 33% of studies after medium and high doses, respectively). Amitriptyline generally impaired central nervous system (CNS) functions, which increased with doses. Multiple doses caused less pronounced effects on the reported tests. The most responsive tests to amitriptyline appeared to be EEG alpha and theta, and REM sleep duration. CONCLUSIONS: SSRIs in healthy subjects appear to cause slight stimulating effects after low doses, which tend to diminish with dose. The most consistent effects were observed with flicker discrimination tests, EEG (alpha and beta bands), REM sleep duration, and subjective effects at higher doses. These effects are small compared with amitriptyline and other CNS-active drugs. Multiple dosing with SSRIs caused even fewer measurable differences from placebo, probably due to adaptive processes. SSRI-effects are best detected with a test battery that is sensitive to general CNS-stimulation, but such tests only comprise a very small portion of the close to 200 different methods that were found in current review.     

LSS法估算COPD患者茶碱缓释片单用及联用加替沙星时稳态药-时曲线下面积

目的用有限采样法（limited sampling strategy,LSS）估算慢性阻塞性肺疾病（COPD）患者单用茶碱缓释片及联用加替沙星时稳态药一时曲线下面积（AUC0-12^ss）。方法测得16名COPD患者口服茶碱缓释片稳态时（单用及联用加替沙星）的血药浓度,以多元线性回归法建立（AUC0-12^ss与不同时间点的血药浓度的关联数学模型,并用Jackknife法进行验证。结果单点预测时可以采用给药后2h或4h的血药浓度值预报药-时曲线下面积,单用茶碱回归方程为：AUC0-12^ss=-3．01＋11．15C2（R^2=0．97,P〈0．05）或AUC0-12^ss=-1．34＋10．91 C4（R^2=0．97,P〈0．05）。联用加替沙星的回归方程：AUC0-12^ss-3．14＋10．27 C2（R^2=0．90,P〈0．05）或AUC0-12^ss=-9．68＋11．74 C4（R^2=0．96,P〈0．05）。二点预测可用给药后2h及12h的血药浓度值,单用茶碱回归方程为：AUC0-12^ss=-0．28＋8．47 C2＋3．18 C12（R^2=0．98,P〈0．05）。联用加替沙星回归方程为：AUC0-12^ss=5．74＋3．64 C2＋8．54 C12（R^2=0．99．P〈0．05）。结论考虑到给药后2h为药物的达峰时间,推荐单点预测时用给药后2h的血药浓度值;用2个浓度点进行预测采用给药后2、12h的血药浓度值。     

Stratification and Partial Ascertainment of Biomarker Value in Biomarker-Driven Clinical Trials

This article examines the role of stratification of treatment assignment with regard to biomarker value in clinical trials that accept biomarker-positive and -negative patients but have a primary objective of evaluating treatment effect separately for the marker-positive subset. It also examines the issue of incomplete ascertainment of biomarker value and how this affects inference about treatment effect for the biomarker-positive subset of patients. I find that stratifying the randomization for the biomarker ensures that all patients will have tissue collected but is not necessary for the validity of inference for the biomarker-positive subset if there is complete ascertainment. If there is not complete ascertainment of biomarker values, it is important to establish that ascertainment is independent of treatment assignment. Having a large proportion of cases with biomarker ascertainment is not necessary for establishing internal validity of the treatment evaluation in biomarker-positive patients; independence of ascertainment and treatment is the important factor. Having a large proportion of cases with biomarker ascertainment makes it more likely that biomarker-positive patients with ascertainment are representative of the biomarker-positive patients in the clinical trial (with and without ascertainment), but since the patients in the clinical trial are a convenience sample of the population of patients potentially eligible for the trial, requiring a large proportion of cases with ascertainment does not facilitate generalizability of conclusions.     

Modeling of drug response in individual subjects

Pharmacokinetic and drug response data from individual subjects are analyzed empirically by two different mathematical techniques. The drugs involved are the antiarrhythmic agent disopyramide, whose kinetics can be described by a two-compartment model, and two cardiac glycosides, digoxin and -methyl digoxin, for which three-compartment models are appropriate. The first analytical approach uses multiple linear regression to describe response in terms of the amount of drug in several kinetic compartments. The second approach describes response in terms of the drug concentration in an effect compartment. Both approaches describe the data equally well and require the same number of parameters for model specification.     

Population pharmacokinetic analysis of clopidogrel in healthy Jordanian subjects with emphasis optimal sampling strategy

Aim: Clopidogrel is metabolized primarily into an inactive carboxyl metabolite (clopidogrel‐IM) or to a lesser extent an active thiol metabolite. A population pharmacokinetic (PK) model was developed using NONMEM® to describe the time course of clopidogrel‐IM in plasma and to design a sparse‐sampling strategy to predict clopidogrel‐IM exposures for use in characterizing anti‐platelet activity.Methods: Serial blood samples from 76 healthy Jordanian subjects administered a single 75 mg oral dose of clopidogrel were collected and assayed for clopidogrel‐IM using reverse phase high performance liquid chromatography. A two‐compartment (2‐CMT) PK model with first‐order absorption and elimination plus an absorption lag‐time was evaluated, as well as a variation of this model designed to mimic enterohepatic recycling (EHC). Optimal PK sampling strategies (OSS) were determined using WinPOPT based upon collection of 3–12 post‐dose samples.Results: A two‐compartment model with EHC provided the best fit and reduced bias in C_max (median prediction error (PE%) of 9.58% versus 12.2%) relative to the basic two‐compartment model, AUC_0‐24 was similar for both models (median PE% = 1.39%). The OSS for fitting the two‐compartment model with EHC required the collection of seven samples (0.25, 1, 2, 4, 5, 6 and 12 h). Reasonably unbiased and precise exposures were obtained when re‐fitting this model to a reduced dataset considering only these sampling times.Conclusions: A two‐compartment model considering EHC best characterized the time course of clopidogrel‐IM in plasma. Use of the suggested OSS will allow for the collection of fewer PK samples when assessing clopidogrel‐IM exposures. Copyright © 2013 John Wiley & Sons, Ltd.     

健康人群乙型肝炎疫苗接种效果评估

目的 了解健康人群乙型肝炎(乙肝)疫苗接种情况,评价乙肝预防效果,为制定免疫策略提供依据.方法 按照两阶段抽样法,在马鞍山市三区一县抽取8个行政村1-59岁常住人口2038人,进行问卷调查,并采集静脉血,用ELISA法检测乙肝血清标志物.率的比较采用卡方检验.结果 乙肝疫苗调查接种率为50.54%,城市高于农村,低年龄组(98.14%)到高年龄组(0.95%)逐渐降低,男性高于女性.散居和托幼儿童接种率最高(96.58%和98.77%),学生次之(87.17%),农民最低(4.26%).1～59岁人群HBsAg阳性率从低年龄组到高年龄组逐渐增高(O%-10.48%),抗-HBs阳性率为14.58%-50.18%,15-岁组最低(14.58%).1990-2007年间乙肝报告发病率以1999年最高,为67.0660/10万,之后呈逐年下降趋势.2005-2007年平均乙肝报告发病率在≤14岁的人群中低于4.6731/10万,在≥15岁的人群中高于32.0789/10万.乙肝血源疫苗接种后HBsAg和抗-HBs阳性率分别为0.98%和39.54%,与基因工程疫苗接种后阳性率(0.49%和43.28%)的差异无统计学意义(x2=0.107,P=0.744;x2=1.004,P=0.316).结论 应继续开展新生儿乙肝疫苗接种,推广成年人接种,并将预防乙肝资源向农村倾斜.