Changing common sense:Anti-platelet/coagulation therapy against cirrhosis

Until recently,anti-platelet/coagulation therapy had not been recommended for patients with cirrhosis.Although venous thrombosis is one of the representative complications of cirrhosis and ischemic disorders associated with atherosclerosis are not infrequent in cirrhotic patients,many clinicians have tended to hesitate to introduce anti-platelet/coagulation therapy to their patients.Undoubtedly,this is due to the increased risk of hemorrhagic diathesis in cirrhotic patients.However,accumulating evidence has revealed the benefits of anti-platelet/coagulation therapy for cirrhotic patients.In addition to the safety of the therapy carried out against cardiovascular diseases in cirrhotic patients,some clinical data have indicated its preventive effect on venous thrombosis.Moreover,the efficacy of antiplatelet/coagulation therapy against cirrhosis itself has been demonstrated both clinically and experimentally.The conceptual basis for application of anti-platelet/coagulation therapy against cirrhosis was constructed through two pathologic studies on intrahepatic thrombosis in cirrhotic livers.It may be better to use thrombopoietinreceptor agonists,which have been tested as a treatment for cirrhosis-related thrombocytopenia,in combination with anti-platelet drugs to reduce the risk of venous thrombosis.During the last decade,the World Journal of Gastroenterology,a sister journal of World Journal of Hepatology,has been one of the main platforms of active discussion of this theme.

Hemorrhagic Bartholin’s cyst in a woman using anti-platelet medication:A case report and review of the literature

BACKGROUND We report the case of a postmenopausal female with a hemorrhagic Bartholin’s cyst who has been using an antiplatelet medication.CASE SUMMARY A postmenopausal woman,84 years of age,had a medical history of hypertension,diabetes mellitus,coronary artery disease(three-vessel disease),chronic kidney disease(stage 3),and dementia.The patient has been taking clopidogrel,an antiplatelet medication,for several years.She presented at our outpatient clinic complaining of painful swelling over her left vulva for several days.A Bartholin’s cyst over the left vulva was suspected,and the patient underwent marsupialization under local anesthesia,which was well-tolerated.During the incision procedure,bright-red blood with some blood clots was discharged,and a hemorrhagic Bartholin’s cyst was observed.There was no recurrence of the hemorrhagic Bartholin’s cyst during the 6-mo subsequent follow-up period.CONCLUSION Hemorrhagic Bartholin’s cysts rarely occur.We report the case of a postmenopausal female with a hemorrhagic Bartholin’s cyst who had been on antiplatelets and was successfully treated with marsupialization.No recurrence was noted during the 6-mo follow-up period.Older females taking antiplatelets should be cautious of bleeding when presenting with a Bartholin’s cyst.

抗血小板溶栓素对局灶性脑缺血/再灌注大鼠脑损伤的作用观察

目的:观察抗血小板溶栓素(anti-platelet thrombolysin,APT)对缺血性脑损伤的保护作用并初步探讨其作用机制。方法:SD大鼠,随机分为假手术组(Sham)、模型组(缺血再灌注组)、阳性组(依达拉奉注射液)、APT高、中、低组。双侧颈动脉结扎,建立脑缺血模型,测定大鼠脑含水量,HE染色观察脑组织病理改变;线栓法建立大鼠脑缺血/再灌注模型,进行行为学评分,酶联免疫法(ELISA)及免疫组化测定脑组织中Toll样受体4(TLR4)、c-Jun氨基末端激酶(JNK)、Bax蛋白含量。结果:与模型组比较,APT高、中剂量组可明显改善缺血再灌注后大鼠神经功能障碍症状,降低脑水肿程度,改善缺血后脑组织的病理改变,明显降低缺血后脑组织中TLR4、JNK、Bax蛋白表达。结论:APT对缺血性脑损伤有较好的保护作用,其机制可能与抑制脑组织中TLR4/JNK/Bax信号通路表达,从而抑制细胞凋亡有关。

抗血小板溶栓素对大鼠脑缺血再灌注损伤保护作用机制研究

目的:探讨抗血小板溶栓素(anti-platelet thrombolysin,APT)对大鼠脑缺血再灌注损伤保护作用的机制。方法:采用TLR4-siRNA在体转染技术,下调大鼠脑组织中Toll样受体4(TLR4)蛋白表达。分别取野生型和TLR4下调的SD大鼠,随机分为假手术组,模型组,TLR4阻断剂(TAK-242 2 mg/kg)组,APT高、中、低组(0. 02、0. 01、0. 005 mg/kg),每组8只。线拴法建立大鼠局灶性脑缺血/再灌注损伤模型,G-LISA法测定大鼠脑组织中RhoA蛋白活性,Western blot法测定脑组织中TLR4、RhoA相关卷曲螺旋形成蛋白激酶(ROCK1/2)、磷酸化的c-Jun氨基末端激酶(pJNK)蛋白表达。结果:与对照组比较,TLR4-siRNA在体转染大鼠脑组织中TLR4蛋白表达明显下降;在野生型大鼠中,与模型组比较,抗血小板溶栓素可明显降低脑组织中TLR4蛋白表达,抑制RhoA活性,减少ROCK1/2、p-JNK蛋白表达;在TLR4下调大鼠中,与模型组比较,APT对脑组织中RhoA活性,ROCK1/2、p-JNK蛋白表达无明显影响。结论:抗血小板溶栓素对大鼠脑缺血再灌注损伤保护作用机制主要与其抑制TLR4/RhoA/ROCK信号通路有关。

溶栓素的纯化研究

目的 :研究溶栓素的纯化工艺。方法 :采用超滤法截留分离 ,强阴离子交换层析去除核酸 ,弱阴/阳离子交换层析和疏水层析组合法精细分离溶栓素。结果 :聚丙烯酰胺凝胶电泳考马斯亮蓝染色法结合显示最终纯化产物为一条染色带 ,纯化倍数为 144 .83,回收率为 38.6 6 %。结论 :溶栓素的纯化工艺基本体现了工业生产工艺的特点 ,具有一定的实用性。

溶栓素的性质及生物活性研究

目的 :研究溶栓素的理化性质、作用机理及其体内溶栓作用。方法 :采用不连续系统 SDS-聚丙烯酰胺凝胶电泳法测定溶栓素的分子量 ,纤维蛋白平板法测定溶栓素的体外活性和研究溶栓素的作用机理 ,不同温度和 p H值下测定体外溶栓作用以确定溶栓素的最适温度和 p H值 ,大鼠颈动脉血栓形成法测定溶栓素的体内溶栓效果。结果 :溶栓素的相对分子量为 30 6 80 ,最适温度是 4 5℃ ,最适 p H值为7.8,溶栓素的作用机理与人血纤溶酶相似 ,有直接纤溶作用 ,溶栓素的体内溶栓作用在同等剂量的条件下优于尿激酶。结论

抗血小板溶栓素对缺糖缺氧再灌注损伤大鼠原代皮质神经细胞保护作用

目的探讨抗血小板溶栓素(APT)对原培养大鼠皮质神经细胞缺糖缺氧损伤的保护作用及其机制。方法大鼠原代皮质神经细胞培养6 d后,经APT 80μg/m L、40μg/m L、20μg/m L、Toll样受体4(TLR4)阻断剂HTA125 10μg/m L预处理24 h,皮质神经细胞缺糖缺氧3 h,恢复正常培养24 h,建立缺糖缺氧再灌注损伤模型。实验分为正常组,模型组,APT低、中、高组,HTA组。倒置显微镜下观察细胞形态,免疫组化染色鉴定神经元。MTT法检测细胞存活率。比色法检测细胞培养液中丙二醛(MDA)含量和乳酸脱氢酶(LDH)活性。G-LISA法检测细胞中Rho蛋白A(Rho A)活性,Western-bloting检测细胞中TLR4、Rho相关卷曲螺旋形成蛋白激酶(ROCK)蛋白表达水平。结果细胞培养6 d后,免疫组化鉴定显示神经细胞纯度达90%以上;与模型组比较,APT低、中、高剂量能明显增加神经细胞存活率;中、高剂量能明显降低培养液中MDA含量和LDH活性,降低细胞中Rho A活性,抑制细胞中TLR4,ROCK1/2蛋白表达。结论 APT对大鼠原代皮质神经细胞缺糖缺氧再灌注损伤有较好的保护作用,其机制与抑制TLR4、Rho A、ROCK信号通路有关。

亲和层析法纯化溶栓素

目的 :探讨亲和层析纯化溶栓素的方法。方法 :采用高碘酸钠法制备溶栓素抗体的亲和层析介质纯化溶栓素。结果 :提纯的溶栓素经聚丙烯酰胺凝胶电泳 ( PAGE)鉴定为一条带 ,活性回收率为 49.5 %。结论 :亲和层析的高碘酸钠法是纯化溶栓素的一种分离速度快、特异性强、经济、安全可靠的方法 。

极性有机溶剂辅助阴、阳离子交换组合色谱法

目的研究极性有机溶剂辅助阴、阳离子交换色谱串接组合纯化蛋白质的可行性。方法以溶栓素为纯化对象 ,采用极性有机溶剂辅助阴、阳离子交换色谱串接组合与常规方法的纯化结果相对比。结果聚丙烯酰胺凝胶电泳考马斯亮蓝染色法结果显示 ,以含低浓度乙醇的缓冲液进行相同的离子交换色谱 ,具有较好的分离效果。结论极性有机溶剂辅助阴、阳离子交换色谱有效串接组合是一种较好的组合纯化方式 ,但是这种方法使用的前提条件 。

溶栓素的分离纯化及特性测定

目的研究一种高效分离纯化溶栓素的方法并测定其pI和相对分子质量(Mr)。方法采用阴、阳离子交换色谱等技术分离纯化溶栓素并通过对缓冲体系的pH值、离子强度的调节及对初始供试品pH值的调节来优化纯化过程,以等电聚焦聚丙烯酰胺凝胶电泳(IEF PAGE)和SDS PHGE方法分别测定纯品的pI和Mr。结果溶栓素达到简捷、高效分离纯化的目的,电泳鉴定为一条蛋白质带,并测定溶栓素的pI为4 .5 0、Mr 为35 10 0。结论溶栓素的纯化工艺达到高纯度纯化程度。

Influences of blood lipids on the occurrence and prognosis of hemorrhagic transformation after acute cerebral infarction: a case-control study of 732 patients

Background: To study the influence of blood lipid levels on hemorrhagic transformation(HT) and prognosis after acute cerebral infarction(ACI).Methods: Patients with ACI within 72 h of symptoms onset between January 1 st, 2015, and December 31 st, 2016, were retrospectively analyzed. Patients were divided into group A(without HT) and group B(HT). The outcomes were assessed after 3 months of disease onset using the modified Rankin Scale(m RS). An m RS score of 0–2 points indicated excellent prognosis, and an m RS score of 3–6 points indicated poor prognosis.Results: A total of 732 patients conformed to the inclusion criteria, including 628 in group A and 104 in group B. The incidence of HT was 14.2%, and the median onset time was 2 d(interquartile range, 1–7 d). The percentages of patients with large infarct size and cortex involvement in group B were 80.8% and 79.8%, respectively, which were both significantly higher than those in group A(28.7 and 33.4%, respectively). The incidence rate of atrial fibrillation(AF) in group B was significantly higher than that in group A(39.4% vs. 13.9%, P<0.001). The adjusted multivariate analysis results showed that large infarct size, cortex involvement and AF were independent risk factors of HT, while total cholesterol(TC) was a protective factor of HT(OR=0.359, 95% CI 0.136–0.944, P=0.038). With every 1 mmol/L reduction in normal TC levels, the risk of HT increased by 64.1%. The mortality and morbidity at 3 months in group B(21.2% and 76.7%, respectively) were both significantly higher than those in group A(8.0% and 42.8%, respectively). The adjusted multivariate analysis results showed that large infarct size(OR=12.178, 95% CI 5.390–27.516, P<0.001) was an independent risk factor of long-term unfavorable outcomes, whereas low-density lipoprotein cholesterol(LDL-C) was a protective factor(OR=0.538, 95% CI 0.300–0.964, P=0.037). With every 1 mmol/L reduction in normal LDL-C levels, the risk of an unfavorable outcome increased by 46.2%. Major therapies, including intravenous recombinant human tissue plasminogen activator(r TPA), intensive lipid-lowering statins and anti-platelets, were not significantly related to either HT or long-term, post-ACI poor prognosis.Conclusions: For patients with large infarct sizes, especially those with cortex involvement, AF, or lower levels of TC, the risk of HT might increase after ACI. The risk of a long-term unfavorable outcome in these patients might increase with a reduction in LDL-C.

Four 14(13→12)‑Abeolanostane Triterpenoids with 6/6/5/6‑Fused Ring System from the Roots of Kadsura coccinea

Four new rearranged 6/6/5/6-fused lanostane-type triterpenoids,kadcoccitanes A-D(1-4),were isolated from the roots of Kadsura coccinea,and their structures were mainly elucidated by comprehensive analysis of their spectroscopic data.Additionally,the structure of 1 was ambiguously verifed by single-crystal X-ray difraction,while the structure of 2,which features a novel 8,16-epoxy motif,was validated by quantum chemical calculation of NMR parameters and ECD spectrum.Moreover,1 and 4 were found to exhibited anticoagulant activity,while 3 and 4 were found to possess anti-platelet aggregation activity.

Novel derivative of Danshensu acts as anti-thrombotic agent for modulation of protein disulfide isomerase family member ERp57

OBJECTIVE To identify the specific targets of a novel derivative of Danshensu ADTM as the protein disulfide isomerase(PDI)family proteins including ERp57.To further investigate the underlying mechanism of ADTM to modulate ERp57 to regulate platelet function with direct interaction withαⅡbβ3 integrin.METHODS To isolate the protein targets that bound to ADTM,a biotin-conjugated ADTM analogue(BAA)was designed and synthesized.BAA(300μmol·L-1)was incubated with rat blood platelet lysates and the BAA-protein complexes were pulled down with NeutrAvidin-agarose followed by protein profiling using LC-MS/MS.To determine platelet aggregation in vitro,rabbit platelets were incubated with the indicated concentrations of compounds and aggregation was induced by ADP(10μmol·L-1)or AA(200μmol·L-1)and measured using a platelet aggregometer.To determine platelet aggregation-induced by ADP in rat in vivo,ADTM(5-20mg·kg-1)in comparison with DSS(10mg·kg-1)and clopidogrel(18mg·kg-1)were administered daily by i.v.injection for 5d,respectively.To determine the action of ADTM on the ERp57/αⅡbβ3 interaction,it was examined by immunoprecipitation with anti-αⅡbβ3antibody,followed by detection of ERp57 immunoreactivity using immunoblotting.RESULTS BAA could bind to various proteins involved in platelet function.In particular,platelet aggregation-associated proteins were identified with>95% protein identification probability including ERp72,ERp57ERp5 and PDI,which are members of the protein disulfide isomerase(PDI)family related to platelet function and redox homeostasis.ADTM exhibited potent inhibition on the redox activity of ERp57 in a concentration-dependent manner(IC50=100 300μmol·L-1).In in vitro studies,ADTM exhibited concentration-dependent inhibition on ADP-induced and AA-induced platelet aggregation with comparable effects to aspirin and clopidogrel.In vivo study showed that ADP-induced platelet aggregation was significantly compromised(>40%reduction)in rats treated with ADTM(20mg·kg-1).Similarly,ADTM also exhibited significant anti-thrombotic effect in vivo as shown in the ferric chloride(FeCl3)-induced venous thrombosis.Immunoprecipitation with anti-αⅡbβ3antibody,followed by detection of ERp57 immunoreactivity using immunoblotting showed that ADTM disrupted the interaction of ERp57 with αⅡ bβ3.CONCLUSION These results demonstrated that ADTM exhibited broad-spectrum anti-platelet activities and ERp57 is a potential therapeutic target for anti-platelet therapy.

Cardiac implications of thrombotic thrombocytopenic purpura

Thrombotic thrombocytopenic purpura(TTP) is a multisystem disorder that essentially can affect any organ in the human body. The hallmark of the pathogenesis in TTP is the large von Willebrand factor multimers on plateletmediated micro-thrombi formation, leading to microvascular thrombosis.Autopsy studies showed that cardiac arrest and myocardial infarction are the most common immediate causes of death in these patients. Clinical manifestations of cardiac involvement in TTP vary dramatically, from asymptomatic elevation of cardiac biomarkers, to heart failure, MI and sudden cardiac death. There is limited knowledge about optimal cardiac evaluation and management in patients with TTP. The absence of typical cardiac symptoms,combined with complicated multi-organ involvement in TTP, may contribute to the under-utilization of cardiac evaluation and treatment. Prompt diagnosis and timely initiation of effective therapy could be critically important in selected cases. Based on our experience and this review of the literature, we developed several recommendations for focused cardiac evaluation for patients with acute TTP:(1) patients with suspected or confirmed TTP should be screened for the potential presence of cardiac involvement with detailed history and physical,electrocardiogram and cardiac enzymes;(2) clinical deterioration of TTP patients warrants immediate cardiac reevaluation;(3) TTP patients with clinical evidence of cardiac involvement should be monitored for telemetry, cardiac biomarkers and evaluated with transthoracic echocardiography. These patients require urgent targeted TTP treatment as well as cardiac-specific treatment. Aspirin therapy is indicated for all TTP patients. Since epicardial coronary artery involvement is rare, cardiac catheterization is usually not required, given the high risk for hemorrhage and kidney injury;(4) we recommend evidence-based medical therapy for ischemic symptoms and heart failure. TTP patients with evidence of cardiac involvement would also benefit from routine cardiology follow up during remission.

Synthesis, activity evaluation and 3D-QSAR study of some novel derivatives of 4, 5, 6, 7-tetrahydrothieno [3,2-c] pyridine

A series of novel derivatives of 4, 5, 6, 7-tetrahydrothieno [3,2-c] pyridine were synthesized and structurally characterized by 1H NMR and MS. Their in vivo anti-platelet aggregation activities were evaluated. A 3D-QSAR was performed using the CoMFA and the CoMSIA. This model provided useful guidelines for novel anti-platelet thienopyridines design.

Association of primary biliary cirrhosis with idiopathic thrombocytopenic purpura

Although both primary biliary cirrhosis (PBC) and idiopathic thrombocytopenic purpura (ITP) are autoimmune diseases, the association of the 2 diseases is rare. Here, we report a case of ITP that developed during the follow-up of PBC in a 74-year- old man. The patient had been diagnosed with PBC 12 years previously, and had received treatment with ursodeoxycholic acid. The platelet count decreased from approximately 60 × 109/L to 8 × 109/L, and the association of decompensated liver cirrhosis (PBC) with ITP was diagnosed. Steroid and immune gamma globulin therapy were successful in increasing the platelet count. Interestingly, human leukocyte antigen genotyping detected the alleles DQB10601 and DRB10803, which are related to both PBC and ITP in Japanese patients. This case suggests common immunogenetic factors might be involved in the development of PBC and ITP.

Adequate antiplatelet regimen in patients on chronic anti-vitamin K treatment undergoing percutaneous coronary intervention

AIM: To investigate the impact of dual antiplatelet therapy (DAT) in patients on antivitamin K (AVK) regimen requiring percutaneous coronary intervention (PCI).METHODS: Between February 2006 and February 2008, 138 consecutive patients under chronic AVK treatment were enrolled in this registry. Of them, 122 received bare metal stent implantation and 16 received drug elutingstent implantation. The duration of DAT, on top of AVK treatment, was decided at the discretion of the clinician. Adequate duration of DAT was def ined according to type of stent implanted and to its clinical indication. RESULTS: The baseline clinical characteristics of patients reflect their high risk, with high incidence of comorbid conditions (Charlson score ≥ 3 in 89% of the patients). At a mean follow-up of 17 ± 11 mo, 22.9% of patients developed a major adverse cardiac event (MACE): 12.6% died from cardiovascular disease and almost 6% had an acute myocardial infarction. Major hemorrhagic events were observed in 7.4%. Adequate DAT was obtained in only 44% of patients. In the multivariate analysis, no adequate DAT and Charlson score were the only independent predictors of MACE (both P = 0.02). CONCLUSION: Patients on chronic AVK therapy represent a high risk population and suffer from a high MACE rate after PCI. An adequate DAT regimen and absence of comorbid conditions are strongly associated with better clinical outcomes.

Chronic hepatitis B： role of anti-platelet therapy in inflammation control

Platelets play a known role in the maintenance of vascular homeostasis, but these cells are emerging as important cellular mediators of acute and chronic inflammatory diseases. Platelets are key elements in the pathogenesis of acute and chronic liver disease associated with hepatitis B virus （HBV） infection by promoting the accumulation of virus-specific CD8＋ T cells and nonspecific inflammatory cells into the liver parenchyma. This review discusses major platelet functions in immune and inflammatory responses, with an emphasis on recent pre-clinical studies that suggest that the inhibition of platelet activation pathways represent an alternative therapeutic strategy with potential use in the reduction of virus-specific T cell-mediated chronic inflammation, liver fibrosis and hepatocellular carcinoma in patients who are chronically infected with HBV.

The value of using polymorphisms in anti-platelet therapy

OBJECTIVES and BACKGROUNDS： Cardiovascular events occure as a result of various risk factors, such as uric acid （UA）, inflammation, hormones and other materials that induce C- reactive protein （CRP） expression. These factors lead to complement activation, and endothelial damages. Damaged endothelial cells release heparan sulfate which inhibits tissue factor activity and von Willed brand factor （VWVF） and causes aggregation. Finally this cascade of events cause platelets aggregation and leads to heart ischemia and cardiovascular events. DISCUSSION： Anti-platelet therapy is an interesting premise. Anti-platelet resistance patients and bleeding as a result of using ticagrelor and prasugrel should be considered in this treatment methods. Anti-platelet drugs such as clopidogrel are prescribed in cardiovascular events. Platelets have VWF receptors and P2Y12 receptors on their surface, and thus, targeting these receptors can be useful in treatment. The active metabolites of clopidogrel bind to P2Y12R and inhibit ADP binding; thus, clopidogrel inhibits aggregation by interfering in several events as a result of the inhibition of ADP attachment to P2Y12R of the platelet. However, the polymorphisms of P2~ 12 and other genes mentioned in Table 1 showed treatment resistance in anti-platelet therapy, highlighting that these SNPs can be helpful in anti-platelet therapy. CONCLUSION： The knowledge of these SNPs may decrease the number of unwanted effects that endanger patients with cardiovascular diseases and avoids ineffective anti-platelet therapy in several patients. Clopidogrel, ticagrelor, prasugrel, and aspirin and CYP2C19 and their SNPs are very important subjects in anti-platelet therapy. To present the importance of using phannacogenetics in anti-platelet therapy, we discuss here the association between these drugs and the SNPs for therapeutic resistance.

The first examples of ilexgenin A hybrids as a new class of multi-potent,anti-platelet agents

Seventeen novel ilexgenin A hybrids（lA-aspirin） and（IA-NO）,as donor hybrids（IA-NO will release NO in vivo and function as NO donor）,were designed and synthesized in order to develop new multi-targeting agents for the treatment of platelet disorders.Their in vitro activities against ADP,AA and thrombin were evaluated.As a result,IA hybrids achieved substantial increases in the three tested pathways compared with IA.Encouragingly,the most potent hybrid compounds 6d and 14d displayed about 8-fold higher potency than aspirin,and 3-fold higher potency than the simultaneous administration of aspirin and IA in inhibiting ADP-induced aggregation with IC_（50） values of 0.15 mmol/L and 0.14 mmol/L,respectively. The results suggest these IA hybrids are good candidates for multi-target therapies,and especially,may be considered as promising ADP agonists.