OBJECTIVE This article is to construct a pharmaceutical care curriculum system under the background of “healthy China”,and to improve the adaptability of pharmaceutical graduates in higher vocational colleges, in order to better serve the society.METHODS Our teaching activities are students-centered, and we adopted a variety of teaching methods and means, built an information-based teaching environment, integrated professional quality and ideological and political education into the teaching process.RESULTS The students′ achievements have been improved, their pharmaceutical service practice skills have been significantly enhanced, and their ideological awareness has been improved.CONCLUSION The reform and practice of pharmaceutical care course can greatly improve the teaching quality of pharmaceutical specialty in Higher Vocational Colleges and enhance the students′ ability to adapt to their future posts.

目的:考察辣椒素对文拉法辛及其活性代谢物O-去甲基文拉法辛体内药动学的影响.方法:选取10只健康SD大鼠,采用自身对照方法,考察单剂量给予盐酸文拉法辛(20.25 mg·kg-1)(对照组)以及连续7d灌胃辣椒素后再给予盐酸文拉法辛(实验组)的药动学差异.采用已验证的液相色谱-串联质谱法(LC-MS/MS)测定血浆中文拉法辛及O-去甲基文拉法辛的浓度,经DAS3.2.4软件计算药动学参数,SPSS软件计算两组实验的差异是否具有显著性.结果:对照组和实验组大鼠血浆中文拉法辛的AUC0-10、Cmax、Tmax、t1/2分别为(780.81±709.76)μg·L-1·h-1和(1037.84±582.63)μg·L-1·h-1、(517.57±462.46)μg·L-1和(876.64±301.79)μg·L-1、(0.32±0.20)h和(0.25±0.09)h、(1.00±0.84)h和(0.89±0.18)h;O-去甲基文拉法辛的AUC0-10、Cmax、Tmax、t1/2分别为(163.60±77.93)g·L-1·h-1和(240.41±62.69)μg·L-1·h-1、(96.83±46.51)μg·L-1和(182.52±46.40) μg·L-1、(0.46±0.30)h和(0.25±0.06)h、(1.19±0.32)h和(2.65±1.58)h;其中实验组的AUC0-10、Cmax均增加,差异具有显著性.结论:辣椒素增加文拉法辛的吸收,提高生物利用度,即能影响文拉法辛的体内药动学过程.

目的 研究在大鼠体内辣椒素对氯氮平及其代谢物的血药浓度及药代动力学的影响.方法 按自身对照方法设计.对照组予以单剂量31.5 mg· kg-1氯氮平,灌胃;经过5d的清洗期后,实验组将对照组大鼠连续灌胃8mg·kg-1辣椒素7d,于第8天灌胃辣椒素(8 mg· kg-1)与氯氮平(31.5 mg· kg-1)混悬液.分别于不同时间点采集血样,并用HPLC-MS/MS法检测氯氮平和去甲氯氮平的血药浓度,用DAS 3.2.4软件计算药代动力学参数.结果 对照组和实验组中氯氮平的主要药代动力学参数如下:Cmax分别为(323.59±97.03)和(560.49±208.53) μg·L-1,tmax分别为(0.95 ±0.35)和(0.85±0.27)h,t1/2分别为(2.13±0.63)和(2.34±0.95)h,AUC0-∞分别为(1769.71±780.02)和(2110.97±1109.88)μg·L-1·h.结论 辣椒素与氯氮平同用时可能会发生药物间相互作用,原因可能是辣椒素会增加氯氮平及其代谢物在大鼠体内的吸收,且抑制氯氮平的消除,因此,临床使用时应注意其潜在影响.

目的:评价高效液相色谱串联质谱法(LC-MS/MS)测定人血清中奥氮平的不确定度.方法:通过分析LC-MS/MS法测定人血清奥氮平浓度过程中的影响因素,包括测量重复性、天平称量、溶液配制、基质效应、血清样本预处理和标准曲线拟合等,评价测定过程中各因素引起的不确定度,计算合成不确定度和扩展不确定度.结果:人血清中低(6 ng·mL-1),中(60ng·mL-1)和高(150 ng·mL-1)浓度奥氮平的扩展不确定度分别为0.418,3.851,10.108 ng·mL-1 (P=95％,k=2).结论:LC-MS/MS法测定人血清中奥氮平的不确定度主要由基质效应和标准曲线拟合引入.

Kynurenine pathway (KP) activation is associated with many neuropsychiatric diseases, such as major depressive disorder (MDD) and Alzheimer's disease (AD). Investigations conducted on MDD seldom shed light on KP changes in late-life depression (LLD), though memory deficit (MD) in patients with LLD is a predictable sign of AD. Thus, we aimed to investigate whether tryptophan (TRP) metabolism and kynurenine (KYN) metabolism were imbalanced in patients with LLD with MD and in patients with LLD without MD. We explored KP characteristics between LLD with MD and LLD without MD groups. We investigated 85 patients with LLD and MD, 71 patients with LLD without MD, and 129 healthy controls (HCs). Serum concentrations of TRP, KYN, and kynurenic acid (KYNA) were detected by liquid chromatography-tandem mass spectrometry. Cognition performance was assessed by the Mini-Mental State Examination (MMSE). Language ability was assessed by the Boston Naming Test (BNT). Depressive symptoms were assessed by the 17-item Hamilton Depression Scale (HAMD-17). Lower TRP and KYNA levels, a lower KYNA/KYN ratio and a higher KYN/TRP ratio were found in patients with LLD and MD compared to those in HC. Low levels of TRP and KYN, in the absence of a changed KYN/TRP ratio, were found in patients with LLD without MD. The KYNA/TRP ratio and MMSE, BNT, and HAMD-17 scores were associated with the presence of LLD. MMSE scores and a trend for the KYN/TRP ratio were associated with the presence of MD in patients with LLD. Aside from MMSE scores, there was a trend toward an association between the KYN/TRP ratio and the presence of MD in patients with LLD. In conclusion, profound shifts in TRP metabolism and KYN metabolism were found in patients with LLD and MD but not in patients with LLD without MD.

OBJECTIVE To compare the results of lamotrigine (LTG) concentration determined by high performance liquid chromatography with diode array detector (HPLC-DAD or HPLC) and mass spectrometer (LC-MS/MS) in serum,and evaluate the correlation and economy of two methods.METHODS Totally 117 serum samples of lamotrigine were detected by HPLC and LC-MS/MS,respectively.Pssing-Bablok regression and Bland-Altman bias plots were used to evaluate the correlation and difference of two methods,and the economy and rationality of two methods were assessed by using the cost-effectiveness analysis method.RESULTS There was no statistically significant difference between the two methods (P =0.707).Passing-Balok regression equation was LTGHPLc =1.074 LTGLcMS/MS + 0.071 and Pearson correlation co-efficient was 0.997.The bias of HPLC for LTG concentration was (0.216 ± 0.347)μg· mL-1,higher than that by LC-MS/MS (95％CI:-0.465-0.897).When daily sample size was ＞9 cases,the profit of HPLC was ＞56.04 yuan.With sample size ＞ 16 cases,the profit of LC MS/MS was ＞36.46 yuan/day,while the profit of HPLC was ＞653.77 yuan/day.When HPLC method achieves maximum sample size (58 cases/day) and the daily size of LC-MS/MS was 66 cases/day,two methods could produce similar daily profit (￥4 240.20 vs 4 305.43).The profit of LC-MS/MS method was ＞5 927.64 yuan/day with sample size ＞ 85 cases,which was far higher than HPLC method.CONCLUSION There is a great correlation between LTG concentrations detected respectively by the two methods.The instrument cost of HPLC is lower than LC-MS/MS method,and HPLC is recommended for the situation of sample size ＜58 cases/day.The detection amount of LC-MS/MS is 3 4 times higher than HPLC method,whose effectiveness is higher for sample size ＞80 cases/day.

Ethnopharmacological relevance: Xiaoyao San (XYS) is a classic Chinese herbal formula for treatment of depression. The present study aimed to investigate the antidepressant effects of XYS in a rat model of chronic unpredictable mild stress (CUMS) and the underlying mechanisms. Materials and methods: A CUMS rat model of depression was established via 4 weeks of unpredictable stimulation. Then the rats were orally administered paroxetine and XYS for 2 weeks with continued stress. Behavioral assessments, including an open field test (OFT), sucrose preference test (SPT) and forced swim test (FST), were conducted to evaluate the antidepressant effects of XYS. The concentrations in rat plasma of tryptophan (Trp) and its metabolic products, including kynurenine (Kyn) and quinolinic acid (QUIN), were determined using high performance liquid chromatography tandem mass spectrometry with electrochemical detection (HPLC-MS/MS). The mRNA and protein levels in rat hippocampus of depression-related brain derived neurotrophic factor (BDNF), cyclic AMP response element binding protein (CREB) and nerve cell adhesion molecule (NCAM) were determined by real-time qPCR and Western blot, respectively. Enzyme Linked Immunosorbent Assay (ELISA) was used to detect the activities of indoleamine 2,3-dioxygenase (IDO) and kynurenine-3-monooxygenase (KMO) in rat plasma. Results: The results showed that a successful CUMS rat model was established through 4 weeks of continuous unpredictable stimulation, as indicated by the significant decrease in locomotor activity and increase in immobility time in the OFT, reduction in body weight and food intake etc. Compared with the normal group, the concentrations of Kyn and QUIN had significantly (p < 0.05) decreased at day 28 in the control group, but then improved after drug treatment with paroxetine and XYS. There were no obvious changes in the activities of IDO and KMO. Compared with the normal group, the mRNA of NCAM, CREB and BDNF were significantly down regulated (p < 0.001) in the control group, BDNF gene was up-regulated by paroxetine or XYS treatment, NCAM and CREB gene did not change in XYS group, protein expressions of BDNF and CREB were significantly increased, and NCAM was significantly reduced (p < 0.05). Conclusions: XYS reversed the abnormalities of the tryptophan-kynurenine metabolic pathways in depressed rats and achieved an excellent antidepressant effect. Its direct impact may be observed as changes in biological indicators in rat hippocampus tissue.

The kynurenine pathway, in which tryptophan is metabolized to kynurenine and kynurenic acid, has been linked to depression. A rapid and highly reproducible liquid-chromatography-tandem mass spectrometry (LC-MS/MS) method were established for determining tryptophan, kynurenine and kynurenic acid in human serum. Biological samples were precipitated with methanol before separation on an Agilent Eclipse XDB-C18. The stable-isotope-labeled internal standards (kynurenine-13C415N and kynurenic acid-d5) were used for quantification. Detection was performed using multiple reaction monitoring in electrospray ionization mode at m/z 205.1→188.1 for tryptophan, m/z 209.1→146.1 for kynurenine, m/z 190.1→144.1 for kynurenic acid. Good linearity of analyte to internal standard peak area ratios was seen in the concentration range 1,000-50,000 ng/mL for tryptophan, 100-5,000 ng/mL for kynurenine and 1-60 ng/mL for kynurenic acid. Pooled drug-free human serum was purified using activated charcoal and the method was shown to be linear, with validation parameters within acceptable limits. The newly developed method was successfully used to determine concentrations of tryptophan, kynurenine and kynurenic acid in serum from 26 healthy volunteers and 54 patients with depression. Concentrations of tryptophan and kynurenine were lower in serum from depressed individuals than from healthy individuals.

目的 建立同时检测抑郁大鼠血浆中文拉法辛及其代谢物O-去甲基文拉法辛的高效液相色谱串联质谱(LC-MS/MS)方法.方法 采用甲醇直接沉淀法处理大鼠血浆样品.色谱柱为Agilent Eclipse plus-C18(4.6 mm× 100 mm,3.5 μm),流动相为甲醇-水(60:40,v/v),流速为0.5 mL·min-1.正离子多反应监测(MRM)扫描分析,离子通道:文拉法辛m/z 278.15→58.20,O-去甲基文拉法辛m/z 264.10→58.10.结果 文拉法辛和O-去甲基文拉法辛的线性范围均为4～400 ng·mL-1,定量下线均为4ng·mL-1,日内、日间精密度RSD均小于11％,相对提取回收率均在90％～ 105％.结论 该方法操作简单,灵敏度高,特异性强,可用于抑郁大鼠血浆内文拉法辛和O-去甲基文拉法辛的含量测定.

目的 考察犬尿氨酸活性物质在健康人与抑郁症患者血清中的含量差异,及抑郁症患者经rTMS治疗后犬尿氨酸活性物质含量变化趋势.方法 收集健康人(26例)及重复经颅磁刺激治疗前后抑郁症患者(54例)的血清;采用LC-MS/MS检测方法测定血清中色氨酸、犬尿氨酸和犬尿喹啉酸的含量.结果 抑郁症患者血清中色氨酸和犬尿氨酸浓度显著低于正常人,其差异有统计学意义(P＜0.002),但犬尿喹啉酸浓度差异无统计学意义(P＞0.05);抑郁症患者经rTMS治疗前后3种物质浓度差异无统计学意义(P＞0.05).结论 健康人与抑郁症患者血清中色氨酸和犬尿氨酸含量有差异,这两物质含量变化可能与抑郁症发病机制有关系.

目的 采用高效液相色谱-串联质谱(LC-MS/MS)法建立同时检测人血浆中奥氮平与拉莫三嗪浓度方法.方法 以Agilent Eclipse plus C18(100 mm× 4.6 mm,5 μm)为分析柱,流动相为含5mmol·L-1甲酸铵的甲醇-水体系(75:25),流速为0.5mL· min-1,以奥氮平-D3及拉莫三嗪-13C3-D3为内标,采用蛋白沉淀法进行样本处理,采用选择性离子监测(MRM),通过三重四级杆串联质谱仪正离子模式同时分析奥氮平(m/z 313→256)与拉莫三嗪(m/z 256→145)血浆浓度,并应用于检测46份临床样本.结果 奥氮平在2～200 μg·L-1(R2 =0.997 2)内线性较好,高、中、低浓度质控日内(n=6)及日间(n=3)精密度RSD值均低于7.13％,准确度在97.47％ ～ 102.70％内,提取回收率为94.24％,103.31％,102.28％;拉莫三嗪在0.2～25 mg·L-1(R2=0.998 8)内线性较好,高、中、低浓度日内(n=6)及日间(n=3)精密度RSD值均低于5.56％,准确度在93.97～102.55％内,提取回收率为100.89％、93.66％、91.27％.结论 本方法可简便、灵敏、高效测定人血浆中奥氮平与拉莫三嗪血药浓度且特异性好,人血浆内源性杂质不干扰奥氮平和拉莫三嗪的测定.

目的 研究老年痴呆伴有高血压患者常用的降压药的临床使用疗效及安全性.方法 共收集老年科2011～ 2014年住院老年人痴呆伴有高血压患者共64例(其中老年性痴呆39例,血管性痴呆15例,混合性痴呆10例),统计入组患者使用的降压药和抗痴呆用药,服药剂量,合并用药、临床疗效及不良反应.结果 老年痴呆伴有高血压患者最主要服用的降压药是钙通道阻滞剂,其中使用氨氯地平(包括左旋氨氯地平)患者占总人数的65.6％.患者服药前后血压明显的降低(P＜0.05)且患者血压得到有效控制,不良反应发生率低.结论 钙通道阻滞剂在老年痴呆伴有高血压患者人群使用较为广泛,且具有疗效显著,安全性高的特点.