A targeted neurotransmitter quantification and nontargeted metabolic profiling method for pharmacometabolomics analysis of olanzapine by using UPLC-HRMS

Neurotransmitters (NTs) are specific endogenous metabolites that act as “messengers” in synaptic transmission and are widely distributed in the central nervous system. Olanzapine (OLZ), a first-line antipsychotic drug, plays a key role in sedation and hypnosis, but, it presents clinical problems with a narrow therapeutic window, large individual differences and serious adverse effects, as well as an unclear mechanism in vivo. Herein, a simultaneous targeted NT quantification and nontargeted metabolomics method was developed and validated for pharmacometabolomics analysis of OLZ by using ultra-high-performance liquid chromatography coupled with high-resolution mass spectrometry (UPLC-HRMS). Considering the low physiological concentrations of NTs, a full MS scan and target selective ion monitoring (tSIM) scan were combined for nontargeted metabolomics and targeted NT quantification, respectively. By using this strategy, NTs at a very low physiological concentration can be accurately detected and quantified in biological samples by tSIM scans. Moreover, simultaneously nontargeted profiling was also achieved by the full MS scan. The newly established UPLC-HRMS method was further used for the pharmacometabolomics study of OLZ. Statistical analysis revealed that tryptophan, 5-hydroxytryptophan, 5-hydroxytryptamine, γ-aminobutyric acid etc. were significantly downregulated, while tyrosine was significantly upregulated, which suggested that OLZ could promote the downstream phase II reaction of 5-hydroxytryptamine, inhibit tyrosine hydroxylase activity, and increase the activity of γ-aminobutyric acid transaminase. In conclusion, this method could provide novel insights for revealing the pharmacodynamic effect and mechanism of antipsychotic drugs.

We developed a method that would provide novel insights for revealing the pharmacodynamic effect and mechanism of antipsychotic drugs (olanzapine).     

手术联合置管冲洗引流病灶治疗脊柱结核的Meta分析

目的：系统分析手术联合置管冲洗引流病灶治疗脊柱结核的治疗效果。方法：两位研究者独立对数据库包括中国知网、万方数据知识服务平台、维普数据库、PubMed、EMbase、Cochrane Library进行文献检索,收集关于手术联合置管冲洗引流病灶治疗脊柱结核的随机对照试验 (RCT) 与回顾性研究。语种限中、英文。两位研...     

Improved detection of human papillomavirus harbored in healthy skin with FAP6085/64 primers

FAP59/64, FAP6085/6319, and CUT primer sets were designed for detecting cutaneous HPV and have been used in many clinical and epidemiology studies. The FAP6085/64 primer set was first evaluated in this study and the FAP6085/64 combination was found to be much more sensitive than all three original primer sets by using HPV plasmids as a template. To confirm further the effectiveness of the FAP6085/64 primer set in human DNA templates, 90 palmar exfoliated cell DNA samples were used to detect the cutaneous HPV by both the FAP59/64 and FAP6085/64 primer sets. The overall proportion of HPV detection in those skin samples was 77.8% (70/90) using FAP6085/64, as compared to 55.6% (50/90) using FAP59/64. The FAP6085/64 primer set was also applied in a population based study. The proportion of HPV detection was 73.96% (2076/2807) in skin samples collected from healthy individuals, and a total of 336 different PV types were found. Sixty (17.9%) of them were fully characterized HPV types, 127 (37.8%) were putative HPV types which had been described previously, 149 (44.3%) were novel putative HPV types, and two animal PVs were also detected. These results suggest that the FAP6085/64 primer set was sensitive and effective for detection of cutaneous HPV in healthy skin samples.     

甘草次酸类化合物的制备和抗肿瘤活性研究

目的制备18α-和18β-甘草次酸类复合物,并对体外抗癌活性进行研究。方法以18β-甘草次酸(Ⅰ)为起点,进行构型转化得到其光学异构体18α-甘草次酸(Ⅱ),再经C30-位羧基的甲酯化分别得到18β-甘草次酸甲酯(Ⅲ)和18-α-甘草次酸甲酯(Ⅳ),并与环磷酰胺的抗癌活性体内代谢产物——二氯磷酰氮芥偶联而得2个抗癌复合物18β-甘甲磷氮芥(Ⅴ)和18α-甘甲磷氮芥(Ⅵ);用四大光谱法(NMR、MS、IR、UV)对各化合物的化学结构进行鉴定分析;利用人类肝癌细胞株BEL-7402为体外实验模型,用MTT法观察各化合物对人肝癌细胞增殖的抑制活性;抗肿瘤药物顺铂作为阳性对照物。结果对化合物的合成工艺进行优化;18α-甘草次酸的构象转化得率为45%;目标化合物18β-甘甲磷氮芥(Ⅴ)和18α-甘甲磷氮芥(Ⅵ)的产率分别为35%和25%。所制备的甘草次酸衍生物中,化合物Ⅱ和Ⅵ对BEL-7402肿瘤细胞的增殖显示明显强的抑制活性,浓度在5～500μg/mL范围内,对肿瘤细胞增殖的抑制率分别为2.6%～86%及1.3%～50.1%;在相同条件下,对照物顺铂的抑制率为20.0%～73.41%。结论目标化合物的制备中磷酰酯化反应的条件控制(无水、物料比1∶1.25,温度65℃和反应时间3～4h)是合成关键;化合物Ⅱ和Ⅵ在中高浓度时,与顺铂具有较类似的抑制肝癌细胞增殖活性。甘草次酸类化合物的构型转化及与抗癌基团偶联可能提高其抗癌潜力。     

Research Article: Binding of Cordycepin Monophosphate to AMP-Activated Protein Kinase and its Effect on AMP-Activated Protein Kinase Activation

It had been reported that cordycepin could activate AMP-activated protein kinase. One possible mechanism is that cordycepin mediated AMP-activated protein kinase activation by conversion into cordycepin monophosphate, which acts as an AMP analog to activate AMP-activated protein kinase. To confirm the aforementioned hypothesis, we investigate the binding of cordycepin monophosphate to AMP-activated protein kinase using molecular docking. The modeling results indicate that cordycepin monophosphate binds to AMP-activated protein kinase with high affinity. The hydrogen bonds provide attractive forces between molecules. Our results further identify the key residues contributing to the interaction. Also, the modeling results predict that cordycepin monophosphate and AMP would have similar binding modes with AMP-activated protein kinase. Further investigation of AMP-activated protein kinase activation in vitro provides the evidence that cordycepin monophosphate functioned as an AMP mimic to activate AMP-activated protein kinase.     

新疆农村地区慢性阻塞性肺疾病的危险因素调查

目的 探讨新疆农村地区COPD危险因素的特点。方法 2007年12月至2010年12月,采用统一的流行病学调查表,以整群随机抽样方法抽取新疆南部和北部农村常住人口3575人为调查对象,进行问卷调查和肺功能检测。以支气管舒张试验后FEV1/FVC＜ 70％,并排除其他心肺疾病作为COPD的诊断标准。采用logistic回归分析方法进行危险因素分析。结果 将资料完整的3489人纳入分析,新疆农村COPD总患病率为4.0％ (138/3489),诊治率仅为14.5％(20/138),维吾尔族的检出率最高(5.1％,90/1774),其次是哈萨克族(3.3％,26/784),汉族的检出率最低(2.4％,22/921),50岁以上人群的COPD检出率明显增高(＞10％,94/743)。COPD患病风险增加的因素有维吾尔族（OR=2.79,95％CI为1.71 ～4.57）、年龄＞30岁（OR =3.41,95％ CI为1.62～7.18）、家族呼吸系统疾病史（OR= 1.68,95％ CI为1.11 ～2.53）和烹饪行为（OR= 1.47,95％CI为1.02 ～2.12）。结论 新疆农村地区COPD患病风险与维吾尔族、年龄、家族呼吸系统疾病史及烹饪行为密切相关。需进一步了解不同民族间COPD患病风险与基因的关系,以及该地区COPD患病风险与生物燃料烟雾的关系。     

In vitro stability and metabolism of O2', O3', O5'-tri-acetyl-N6-(3-hydroxylaniline) adenosine in rat, dog and human plasma: chemical hydrolysis and role of plasma esterases

O2', O3', O5'-tri-acetyl-N(6)-(3-hydroxylaniline)adenosine (WS070117), a new structure-type lipid regulator, is being developed in pre-clinical study. In order to monitor drug kinetics it is essential to understand pre-analytical factors that may affect drug assay. In vitro stability and metabolism were investigated using high-performance liquid chromatography (HPLC) method in this study. The hydrolysis products were identified by HPLC-mass spectrometry (MS)/MS method. The esterases involved in WS070117 hydrolysis was assigned via inhibition rate assay. It was found that WS070117 was chemically unstable in alkaline solutions compared to acidic and near neutral solutions. Enzymatic hydrolysis was even more rapid. Hydrolytic rate constants differ between species, being 4.24, 5.96?×?10(-3) and 6.85?×?10(-2) min(-1) in rat, dog and human plasma at 37°C, respectively. The hydrolysis was catalyzed by plasma esterase because NaF (sodium fluoride: a general esterase inhibitor) inhibited WS070117 hydrolysis and metabolite production. Hydrolysis was fast in rat plasma and was catalysed by carboxylesterase and butyrylcholinesterase. In dog plasma, carboxylesterase, butyrylcholinesterase and paraoxonase were mainly responsible. Butyrylcholinesterase was the major esterase involved in WS070117 hydrolysis in human plasma. The WS070117 hydrolysis in plasma proceeded by gradual loss of acetyl groups. The knowledge of in vitro drug stability and metabolic pathways identified in this study will be essential for future pre-clinical and clinical pharmacokinetics studies.     

Effect of Cydonia oblonga Mill. fruit and leaf extracts on blood pressure and blood rheology in renal hypertensive rats

Ethnopharmacological relevance

Quince, Cydonia oblonga Mill. (COM), is used in traditional Uyghur medicine to treat or prevent cardiovascular diseases. Uyghur people have greater longevity and lower blood pressure than other central Asian populations. We therefore tested COM fruit and leaf extracts on blood pressure and rheology in renal hypertensive rats (RHR).

Materials and methods

Two-kidney, one-clip (2K1C) renal hypertensive rats were divided randomly into eleven groups: sham, model, and model treated with daily doses of 80 and 160 mg/kg aqueous or ethanol extracts of COM fruit or leaves, or 25 mg/kg captopril (n=10 per group), given orally once daily for 8 weeks. Blood pressure was measured before treatment and every 2 weeks thereafter. Blood rheology was tested after 8 weeks.

Results

Model rats had higher blood pressure than sham 8 weeks after the procedure (systolic blood pressure 193±7 vs. 138±8 mmHg, p<0.05). Those treated with captopril had decreased blood pressure within 2 weeks but that did not return to the level found in the sham group at 8 weeks (167±7, p<0.05 vs. model). With the COM extracts, the effect on blood pressure was notable after 4 weeks. At 8 weeks blood pressure was similar with captopril and with 160 mg ethanol leaf extract (166±4, p<0.05 vs. model), the most effective of the extracts. Model rats had higher blood viscosity and lower erythrocyte deformability than sham. Captopril had little effect on blood rheology; whereas COM extracts reduced whole blood viscosity and improved erythrocyte deformability to levels approaching those found in sham.

Conclusions

COM extracts have antihypertensive activity in renal hypertensive rats. The additional effect on rheology, compared to captopril, may convey added interest. Further studies of these effects in man appear warranted.