大黄中有效成分提取分离条件的优化

目的 :优化从大黄中提取分离大黄蒽醌类有效成分的条件 ,以提高其收率和纯度。方法 :用乙醇回流提取大黄 4次 ,得到粗提取液后 ,用硼酸盐缓冲溶液萃取 ,萃取液经高压制备色谱分离纯化 ,得大黄酸、大黄素、大黄酚单体成分 ,丙酮重结晶后得纯品。色谱柱为YWGC18柱 (4 .6mm× 2 5 0mm ,10 μm)流动相为甲醇 水 高氯酸 (9 1 0 .0 1) ,流速 1.0ml·min-1,检测波长 2 5 4nm。结果 :用高压制备系统可分离纯化大黄蒽醌类有效成分 ,得到大黄酸、大黄素、大黄酚单体成分 ,各成分纯度在 80 %以上 ,大黄素收率达 0 .6%。结论 :醇提法提取大黄蒽醌类有效成分的方法简单 ,提取率高。高压制备系统可分离纯化大黄蒽醌类有效成分 ,得单体纯品 ,且收率高纯度好。     

The genetic basis of antiplatelet and anticoagulant therapy: A pharmacogenetic review of newer antiplatelets (clopidogrel,prasugrel and ticagrelor) and anticoagulants (dabigatran,rivaroxaban, apixaban and edoxaban)

Introduction: The study of pharmacogenomics presents the possibility of individualised optimisation of drug therapy tailored to each patients’ unique physiological traits. Both antiplatelet and anticoagulant drugs play a key role in the management of cardiovascular disease. Despite their importance, there is a substantial volume of literature to suggest marked person-to-person variability in their effect.

Areas covered: This article reviews the data available for the genetic cause for this inter-patient variability of antiplatelet and anticoagulant drugs. The genetic basis for traditional antiplatelets (i.e. aspirin) is compared with the newly available antiplatelet medicines (clopidogrel, prasugrel and ticagrelor). Similarly, the pharmacogenetics of warfarin is compared with the newer direct oral anticoagulants (DOACs) in detail.

Expert Opinion: We identify strengths and weaknesses in the research thus far; including shortcomings in trial design and a review of newer analytical techniques. The direction of this research and its real-world implications are discussed.     

Investigational drugs for the treatment of acute myocardial infarction: focus on antiplatelet and anticoagulant agents

Background: Advances in our understanding of the complex pathophysiologic mechanisms responsible for high-risk atherosclerotic plaque rupture resulting in acute myocardial infarction (AMI) have led to the development of numerous antiplatelet and anticoagulant agents for treatment of AMI.

Areas covered: We review various antithrombotic drugs which were recently investigated for the treatment of AMI. A MEDLINE search for relevant articles on newer antiplatelet agents and anticoagulants drugs for the treatment of AMI was performed, and important original investigations were reviewed. We also briefly discuss agents that completed evaluation and were recently recommended by expert guidelines.

Expert opinion: The antiplatelet agents cangrelor and vorapaxar and the anticoagulant rivaroxaban, have shown promise for the reduction of ischemic events when administered during, and in the acute phase following AMI. However, these agents have not been compared with more potent P2Y₁₂ inhibitors, prasugrel, and ticagrelor. Finding an optimum combination of these agents to achieve an appropriate risk (bleeding) – benefit (reduction in ischemic events) balance is challenging. Further evaluation of agents that show promise is important for enhancing our armamentarium of pharmacologic agents for the successful treatment of AMI.     

煎煮方法对生大黄主要成分含量的影响

目的:探讨不同煎煮方法和煎煮时间对生大黄煎液中主要蒽醌类成分及没食子酸含量的影响。方法:用高效液相色谱法测定生大黄煎液中总的和游离的芦荟大黄素、大黄酸、大黄素、大黄酚、大黄素甲醚及没食子酸含量。色谱柱:Diamonsil C18(250mm×4.6mm,5μm);柱温30℃;流速1mL.min-1;测定蒽醌类成分流动相为甲醇-0.1%磷酸(83∶17),检测波长254nm;测定没食子酸流动相为甲醇-0.01%磷酸(10∶90),检测波长273nm。结果:采用后下和正常煎煮方法,在10～60min内,时间越长,各游离或结合蒽醌衍生物及没食子酸的含量均有不同程度增加。结论:大黄煎液中游离或结合蒽醌含量与泻下作用无直接量效关系。     

Clinical trials of an antiplatelet agent,ticlopidine, in diabetes mellitus

Summary

At present there is no simple, reliable and noninvasive method for monitoring progression and improvement in diabetic microangiopathy. However; some diabetic patients with severe microvascular complications show a fairly specific pattern of impaired left ventricular function (abnormal relaxation, cavity filling and wall thinning) and abnormalities of haemorheology (increased viscosity, erythrocyte rigidity and beta-thromboglobulin and decreased threshold for platelet ADP aggregation). A single-blind, 6-months’ crossover study of an antiplatelet agent, ticlopidine, was conducted in 20 diabetics with clinical evidence of microvascular disease. Response to therapy was monitored by digitised M-mode echocardio-graphic analysis of left ventricular diastolic function and haemorheology. All patients had abnormal basal values with no significant change during the 3-month placebo run-in period but, although significant alterations in viscosity, erythrocyte deformability, beta-thromboglobulins and ADP threshold were observed, no change in left ventricular function was detected. It is concluded that, while it may be possible to alter abnormal haemorheology in diabetes, there was no change in one parameter of microvascular end-organ damage.     

CX-659S: a novel diaminouracil derivative that has antioxidative and acute anti-inflammatory activities

We investigated the antioxidative activities and the effects on acute inflammation in mice of a novel diaminouracil derivative, CX-659S ((S)-6-amino-5-(6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxamido)-3-methyl-1-phenyl-2,4(1H,3H)-pyrimidinedione). CX-659S showed potent scavenging activities against the hydroxyl radical and peroxynitrite and inhibited lipid peroxidation in rat brain homogenates in vitro. Topically applied CX-659S dose-dependently inhibited arachidonic acid- and 12-O-tetradecanoylphorbol 13-acetate (TPA)-induced ear edema in mice. Consistent with its antioxidative properties in vitro, CX-659S dramatically attenuated the accumulation of lipid peroxides in the mouse ear elicited by repeated application of TPA. Previously, we reported the effectiveness of CX-659S against contact hypersensitivity reactions in both mouse and guinea pig models. These present results further suggest the therapeutic potential of CX-659S for acute skin inflammation that may involve oxidative tissue damage.     

海藻多糖UH3的结构特征、抗凝和溶栓活性研究

目的 对海藻多糖UH3的结构特征、抗凝血和溶栓活性进行研究。方法 通过热水提取法，从海藻中提取硫酸多糖，采用强阴离子交换色谱和凝胶渗透色谱对多糖进行分离纯化；采用高效液相色谱、高效凝胶渗透色谱（HPGPC）、红外光谱及甲基化方法对多糖的结构进行表征；通过活化部分凝血活酶时间（APTT）、凝血酶时间（TT）、凝血酶原时间（PT）及血凝块溶解法研究多糖体外抗凝血和溶栓活性。结果 多糖UH3在HPGPC色谱图上呈单一对称峰，分子量为50.3 kDa，硫酸根和糖醛酸含量分别为12.27和12.85%；UH3主要由鼠李糖组成，含有少量葡萄糖醛酸和木糖；UH3糖链中鼠李糖主要以→4)-Rhap-(1→、 →3,4)-Rhap-(1→及少量的T-Rhap形式存在，木糖以→4)-Xylp-(1→和T-Xylp存在形式，糖醛酸以→4)-GlcAp-(1→和T-GlcAp形式存在；硫酸根主要位于→4)-Rhap-(1→的C-3位。多糖UH3对APTT和TT有显著延长作用，并能明显提高溶栓率。结论 海藻多糖UH3是一种结构新颖的具有抗凝和溶栓活性的硫酸多糖。     

HPLC法测定复方奥硝唑大黄口腔膜中的大黄酸、大黄素及大黄酚的含量

目的建立反相高效液相色谱法(RP-HPLC)测定复方奥硝唑大黄口腔膜中大黄酸、大黄素及大黄酚的含量。方法使用Fortis Xi C18色谱柱(250 mm×4.6 mm,5μm),甲醇-质量分数为0.1%的磷酸溶液(体积比为85∶15)为流动相,流速为1.0 mL.min-1,检测波长为254 nm。结果大黄酸、大黄素、大黄酚峰与相邻色谱峰分离度均高于3.0,方法专属性强;各组分在测定范围内线性关系良好,回归方程分别为:大黄酚A大黄酚=3.298 3×107ρ-602 4(R2=0.999 0),大黄酸A大黄酸=2.138 3×107ρ-576 2(R2=0.997 0),大黄素A大黄素=2.075 0×107ρ-489 1(R2=0.9990);各组分加样回收率均高于94.0%。测得复方奥硝唑大黄口腔膜中大黄酸、大黄素、大黄酚的平均含量为353、106、121μg.cm-2。结论本方法测定奥硝唑大黄口腔膜中大黄酸、大黄素及大黄酚,可应用于奥硝唑大黄口腔缓释膜剂的质量控制。     

生熟大黄总提取物灌胃给药后游离蒽醌在SD大鼠组织中的分布差异研究

大黄应用范围广、疗效确切,然而近年来国内外实验报道大黄中蒽醌成分具有肝、肾细胞毒性。本文对比研究了生熟大黄总提取物灌胃给药后游离蒽醌在正常SD大鼠组织中的分布。肝、脾和肾组织中游离蒽醌类成分含量采用UPLC-MS/MS法测定。生熟大黄总提物(相当于生药量14.69 g.kg-1)灌胃给药,连续12周。结果表明,游离蒽醌在生大黄组大鼠组织中的分布浓度高于熟大黄组;大黄游离蒽醌在同一组织中的分布浓度顺序大致为大黄酸>大黄素>芦荟大黄素;大黄酸在肝、脾和肾组织中的分布浓度明显高于芦荟大黄素和大黄素,且大黄酸在生大黄组的组织分布浓度明显高于熟大黄组;停药恢复4周后,大黄游离蒽醌在组织中检测不到。结果提示生大黄的组织毒性可能高于熟大黄;大黄酸可能是大黄主要的毒性物质之一;大黄游离蒽醌在组织中可能无蓄积毒性。炮制过程不仅影响了有效成分的含量还可能影响其成分的组成,通过影响吸收和作用过程中成分之间的相互作用,致使生大黄组动物组织中游离蒽醌的分布浓度高于熟大黄组,这与传统中医药理论炮制减毒的思想认识基本一致。     

萱藻多糖的制备及抗凝血抗血栓活性研究

目的制备萱藻多糖并对家兔体外凝血及小鼠体内血栓形成的抑制作用进行研究。方法采用热水浸提-乙醇沉淀法制备萱藻多糖;检测家兔心脏动脉血活化部分凝血酶时间(APTT),凝血酶原时间(PT)和凝血酶时间(TT)。以角叉菜胶诱导小鼠尾部血栓模型,检测萱藻多糖体内抗血栓作用。结果萱藻多糖可显著延长家兔血浆APTT、PT和TT;对角叉菜胶诱导的小鼠尾部血栓具有显著的抑制作用(P<0.01)。结论萱藻多糖具有抗凝和抑制实验性血栓形成的作用。     

沙蚕(Perinereis aibuhitensis)硫酸多糖的结构表征及抗凝血活性研究

目的 对来源于沙蚕（Perinereis aibuhitensis）的硫酸多糖进行结构表征和抗凝血活性研究。方法 采用两步酶解法从沙蚕中提取多糖;利用强阴离子交换色谱和凝胶渗透色谱对粗多糖进行分离纯化;通过离子色谱法、高效液相色谱法（HPLC）、高效凝胶渗透色谱-多角度激光光散射仪（HPGPC-MALLs）联用技术、硫酸软骨素酶ABC酶法分析、核磁共振氢谱（1H-NMR）等方法,研究纯化多糖的化学组成和结构特征;通过测定APTT、PT和TT评价其体外抗凝血活性。结果 从沙蚕中纯化得到了2种硫酸多糖组分PAE1和PAE2,二者的总糖含量、蛋白含量、硫酸根含量及分子量分别为65.21%、14.31%、0.33%、24.49 kDa和53.08%、11.33%、13.46%、57.39 kDa。PAE1主要由Gal（43.58%）、Glc（32.63%）、GalN（8.71%）、GlcA（7.66%）及少量Fuc（2.77%）组成;PAE2主链为硫酸软骨素C（GlcAβ1→3GalNAc6S）,支链主要由Fuc（35.33%）、Gal（20.9%）和Glc（8.98%）构成。PAE1和PAE2均可明显延长APTT和PT。结论 首次从沙蚕中提取、分离得到2种硫酸多糖,其中PAE2是1种含有Fuc、Gal与Glc支链并具有明显的体外抗凝血活性的结构新颖的类硫酸软骨素,该发现为沙蚕硫酸多糖的开发提供了依据。     

大黄总蒽醌的提取精制工艺研究

目的:优选大黄总蒽醌的提取精制工艺。方法:以大黄总蒽醌的转移率为指标,采用单因素法确定提取工艺,通过正交试验确定阴离子交换树脂精制大黄总蒽醌的工艺。结果:最终确定的工艺为大黄药材用10倍量的30%乙醇浸泡,加热回流提取3次,时间分别为1,0.5,0.5 h,合并提取液,减压回收乙醇得浓缩液,将浓缩液用4% NaOH调pH到10,过处理好的阴离子交换树脂柱,用去离子水洗至流出液呈中性,用2 mol·L^-1盐酸酸化,用10倍量体积的85%以上乙醇解析,回收乙醇,将析出的沉淀过滤,50 ℃以下干燥,得黄棕色粉末,总蒽醌含量54.5%,转移率34.6%。结论:制备得到的总蒽醌含量较高,采用阴离子交换树脂提取纯化大黄总蒽醌的工艺可行。     

Triple antiplatelet therapy vs. dual antiplatelet therapy in patients undergoing percutaneous coronary intervention: an evidence-based approach to answering a clinical query

AIMS

Outcomes of patients undergoing percutaneous coronary intervention (PCI) with drug-eluting stents (DES) and bare metal stents (BMS) have not been evaluated separately for specific dual and triple antiplatelet agent use. The purpose of this meta-analysis was to determine whether triple antiplatelet therapy (combination of clopidogrel, aspirin and cilostazol) has any advantage in efficacy compared with standard dual antiplatelet therapy (aspirin and clopidogrel) in patients undergoing PCI.

METHODS

Electronic and printed sources were searched till May 2008 for randomized controlled clinical trials (RCTs) of cilostazol in combination with aspirin and clopidogrel. Pooled weighted mean difference (WMD) and pooled odds ratio (OR) with 95% confidence intervals (CIs) were calculated.

RESULTS

A total of four RCTs including 1457 patients with a median follow-up period of 6–9 months were included in the analysis. The rates of major adverse cardiac and/or cerebrovascular events (MACE/MACCE), stent thrombosis and bleeding were not significantly different between triple and dual antiplatelet therapy groups. Pooled analysis showed that cilostazol was associated with significantly decreased incidence of in segment restenosis (ISR) (OR 0.51, 95% CI 0.38, 0.68; P < 0.00001), increased minimum luminal diameter (MLD) (WMD 0.16, 95% CI 0.10, 0.22; P < 0.00001) for both DES and BMS and also individually. However, the rates of target vessel revascularization (OR 0.45, 95% CI 0.25, 0.83; P = 0.01 and late lumen loss (pooled WMD 0.14, 95% CI 0.2, 0.07; P = 0.001) were decreased significantly only in the DES group receiving triple therapy.

CONCLUSIONS

Cilostazol appears to be effective in reducing the rates of ISR without any significant benefit for MACE/MACCE.     

Antiplatelet and antithrombotic activities of timosaponin B-II, an extract of Anemarrhena asphodeloides

1. Antithrombotic agents are effective in the treatment of ischaemic stroke. Timosaponin B-II (TB-II) is a major active component of Anemarrhena asphodeloides Bunge (Liliaceae; rhizome) that has protective effects against cerebral ischaemic damage. The present study examined the antiplatelet and antithrombotic actions of TB-II. 2. In in vitro experiments, TB-II (20, 40 and 80 mg/mL) potently and dose-dependently inhibited ADP-induced platelet aggregation. Furthermore, 1, 3 and 6 mg/kg TB-II prolonged activated partial thromboplastin time by 9.29, 16.86 and 25.50%, respectively, but had no effect on the prothrombin time. Furthermore, 1, 3 and 6 mg/kg TB-II significantly reduced the wet weight, dry weight and length of the thrombi (%inhibition (based on wet weight): 13.6, 19.8 and 24.7%, respectively). 3. In a rabbit arteriovenous shunt model, 1, 3 and 6 mg/kg, i.v., TB-II had no effect on thrombus formation. Plasma euglobulin lysis time and fibrin degradation product were not affected by 1, 3 and 6 mg/kg TB-II, but plasminogen levels were decreased significantly by 14.4, 18.3 and 29.0%, respectively. 4. The results of the present study demonstrate significant antiplatelet and anticoagulation effects of TB-II and suggest that these actions could contribute to its neuroprotective effect against damage following cerebral ischaemia damage.     

生、熟大黄及其在下瘀血汤中对热结血瘀模型大鼠血液流变学的影响

目的 比较生、熟大黄及其入下瘀血汤对热结血瘀模型大鼠的血液流变学的影响,以阐释生大黄泻热通便、熟大黄活血化瘀的炮制作用。方法 采用ig热性中药结合sc盐酸肾上腺素的方法,复制大鼠热结血瘀模型,再ig不同剂量的生、熟大黄以及生、熟大黄组成下瘀血汤,观察对模型大鼠血液流变学的影响。结果 熟大黄各剂量组与等剂量的生大黄相比,其血液流变学的各项指标检测值均有所改变,其中熟大黄高剂量组全血黏度、红细胞刚性指数与变形指数（TK）与生大黄相比,差异具有显著性（P＜0.01）;熟大黄复方组与等剂量生大黄复方组比较,其血液流变学的各项指标也有明显改变。结论 熟大黄的活血化瘀作用强于生大黄,大黄炮制后可增强其活血化瘀作用。     

大黄中大黄酚-8-O-β-D-吡喃葡萄糖苷的提取、纯化及含量测定方法的研究

目的对大黄中大黄酚-8-O-β-D-吡喃葡萄糖苷进行提取、纯化及含量测定的方法进行研究。方法采用大孔吸附树脂对大黄酚-8-O-β-D-吡喃葡萄糖苷进行纯化,采用高效液相色谱法,以自制大黄酚-8-O-β-D-吡喃葡萄糖苷标准品作对照,对大黄药材中有效成分大黄酚-8-O-β-D-吡喃葡萄糖苷进行了含量测定。色谱柱：Kromasil C18（250mm×4.6mm,5μm）;柱温：40℃;流动相∶乙腈-水（30∶70,v/v）;流速：1.0mL·min-1;检测波长：420nm。结果被测峰和其他峰可完全分离,在10～200μg·mL-1内具有良好的线性关系,r=0.9998,测得药用大黄中大黄酚-8-O-β-D-吡喃葡萄糖苷含量为0.713%,回收率：97.82%,RSD：0.84%。结论这种方法准确、可靠,可用于大黄药材的质量控制。     

Developing drugs for use before,during and soon after percutaneous coronary intervention

Introduction: Percutaneous coronary intervention (PCI) is a milestone for treating coronary artery disease (CAD). Antithrombotic therapy is essential to prevent ischemic complications, including the microvascular no-reflow, while minimizing bleeding events.

Areas covered: This overview discusses available and developing drugs for PCI including anticoagulants, antiplatelets and treatment of no-reflow.

Expert opinion: For years unfractionated heparin (UFH) has been the unique anticoagulant to be used before and during PCI. Enoxaparin showed similar efficacy and safety, yet, based on recent trials, bivalirudin has been shown to have some benefits, particularly for patients with ST-segment elevation myocardial infarction (STEMI). The evidence concerning new anticoagulants is still preliminary, except for new oral anticoagulants, particularly rivaroxaban that showed intriguing findings and is currently under investigation. Dual antiplatelet therapy (DAPT) is the standard of care after PCI, but new developments have recently emerged. Indeed, ticagrelor and prasugrel are currently recommended over clopidogrel due to their significant reduction of ischemic events in acute coronary syndrome (ACS) whereas clopidogrel remains the choice in stable CAD. Among new agents, vorapaxar and cangrelor showed positive but limited evidence and might be considered at least in selected patients. Conversely, evidence on effective treatments for no-reflow remains limited and would require future dedicated research.