Evaluation of the relationship between the platelet reactivity detected by two methods and CYP2C19 gene polymorphism in patients with ACS after receiving clopidogrel / 解放军医学杂志

Objective To evaluate the relationship between the platelet reactivity detected by two methods and CYP2C19 gene polymorphism in patients with ACS after taking clopidogrel, and analyze the correlation between different platelet detection methods. Methods Forty-three patients with ACS, admitted in the General Hospital of Northern Theater Command from July to October 2019, met the diagnostic criteria and received dual antiplatelet therapy, were enrolled in present study. The gene chip was used for CYP2C19 genotype detection, vasodilatory stimulated phosphorylated protein (VASP) method and two revulsiveinduced phototurbidimetric methods [arachidonic acid-induced phototurbidimetry (AA-LTA), diphosphate adenosine-induced phototurbidimetry (ADP-LTA)] were used to evaluate the antiplatelet reactivity, and the correlation between different evaluation methods was compared. Results Of the 43 patients, 17(39.5%) were EMs, 19(44.2%) were IMs, and 7(16.3%) were PMs. The results of VASP were higher in PMs and IMs patients than those in EMs, and the results of AA-LTA were higher in PMs than those in IMs and EMs. The differences were significant (P<0.01). The platelet reactivity of VASP was positively correlated with AA-LTA and ADP-LTA (r=0.383, 0.369, P<0.05); and AA-LTA was positively correlated with ADP-LTA (r=0.496, P=0.011). Conclusions There is a difference between CYP2C19 gene polymorphism and platelet reactivity, and the correlation between VASP method and LTA method is low. CYP2C19 genotype can be used as a reference for the platelet reactivity of patients after clopidogrel therapy.

Effect of CYP2C19 polymorphism on platelet P2Y12 receptor signaling pathway in Chinese patients with stable coronary artery disease after the treatment of clopidogrel / 复旦学报（医学版）

Objective To investigate the molecular mechanism of the effect of CYP2C19 polymorphism on the efficacy of clopidogrel in patients with stable coronary artery disease (SCAD).Methods A total of 208 patients with coronary artery disease undergoing percutaneous coronary intervention (PCI) were included.The CYP2C19 variant alleles were detected by gene sequencing.Platelet reactivity was assessed by thrombelastograph at 24 h after 300 mg clopidogrel loading.P2Y12-Gi signaling was measured by flow cytometric analysis of vasodilator-stimulated phosphorylation-platelet reactivity index (VASP-PRI) and Akt phosphorylation (P-Akt) index.Results Among 208 patients,40.9％ were classified as CYP2C19 * 1/* 1 (non-carriers) with no loss-of-function (LOF),44.7％ as CYP2C19 * 1/* 2 or CYP2C19 * 1/* 3 (one LOF carriers) and 14.4％ as CYP2C19 * 2/*2 or CYP2C19 * 2/* 3 (two LOF carriers).Both postclopidogrel platelet aggregation and VASP-PRI increased by the presence of one LOF allele,and were even more pronounced with the presence of two LOF alleles.In contrast,P-Akt index only increased in two LOF carriers.VASP-PRI was positively associated with postclopidogrel platelet aggregation (r =0.661,P＜0.001),but not with P-Akt index.Conclusions The two CYP2C19 LOF carriage was associated with more active state of P2Y12-Gi signaling in postclopidogrel platelet in Chinese patients with SCAD.

Evaluation of clopidogrel response variability and identification of the CYP2C19 polymorphism in Mexican patients / Evaluación de la variabilidad en la respuesta a clopidogrel e identificación del polimorfismo CYP2C19 en pacientes mexicanos

Abstract Objective Drug inhibition of platelet P2Y12 adenosine diphosphate receptor has reduced the incidence of adverse cardiovascular events after percutaneous coronary interventions. The analysis of the phosphorylation status of vasodilator-stimulated phosphoprotein by flow cytometry has shown a predictive value for adverse events and stent thrombosis. Polymorphisms of CYP2C19 in high risk patients may also relate to adverse cardiovascular events. Methods Ninety patients were enrolled. Patients received a 600 mg clopidogrel loading dose. Blood samples were obtained at the time of the procedure and 24 h later, platelet reactivity was assessed by vasodilator-stimulated phosphoprotein phosphorylation measurement using flow cytometry. Low response to clopidogrel was defined as a platelet reactivity index ≥ 50%. The presence of CYP2C19*2 was identified with the restriction enzyme Smal. Results Mean platelet reactivity index: 53.45 ± 22.48% in the baseline sample and 57.14 ± 23.08% at 24 h (p = 0.183); 40% of patients behaved as good responders, the rest behaved as non-responders with 38% of patients showing platelet reactivity indexes between 50-70% and 22% showing indexes above 70%. The CYP2C19*2 polymorphism was found in 17% of patients, with a 3.9% AA homozygous genotype carriers. Conclusion Response to the clopidogrel loading dose showed a wide variability among patients with 40% responding to the drug according to previously established cut-off values. Our results showed that 3.9% of patients show the AA genotype. To our knowledge, this is the first study involving clopidogrel response by flow citometry and genotype typification in Mexican Mestizo population.

Resumen Objetivo La inhibición del receptor plaquetario P2Y12 se ha asociado con reducción en incidencia de eventos cardiovasculares mayores en pacientes sometidos a intervenciones coronarias percutáneas. El estudio de la fosfoproteína estimulada por vasodilatadores mediante citometría de flujo tiene valor predictivo para desarrollo de eventos adversos y trombosis del stent. Los polimorfismos del CYP2C19 en pacientes de alto riesgo pueden también asociarse con eventos adversos. Método 90 pacientes, dosis de carga de clopidogrel: 600 mg. Se obtuvieron muestras de sangre basales y post-24 horas. La reactividad plaquetaria se estudió mediante medición de fosfoproteína estimulada por vasodiatadores por citometría de flujo. Se consideró baja respuesta al clopidogrel un índice de reactividad plaquetaria ≥50%. La presencia del CYP2C19*2 se identificó con enzima de restricción Smal. Resultados La media del índice de reactividad plaquetaria fue: 53.45 ± 22.48% en muestras basales y 57.14 ± 23.08% a 24 h (p = 0.183); 40% de los pacientes repondieron a clopidogrel, el resto de comportó como no-respondedores, un 38%, mostró índices de reactividad plaquetaria entre 50 -70% y 22%, índices > 70%. El polimorfismo CYP2C19*2 se encontró en 17% pacientes, con un 3.9% portadores de genotipo homozigótico AA. Conclusiones La respuesta a clopidogrel mostró amplia variabilidad entre pacientes, el 40% presentó respuesta de acuerdo con puntos de corte pre establecidos. Un 3.9% de los pacientes presentó genotipo AA. Consideramos que este es el primer estudio realizado en población mestizo-mexicana utilizado citometría de flujo para evaluar la respuesta a clopidogrel así como la tipificación genética de los pacientes.

The Relationship of VASP Signaling Molecule and Colorectal Cancer / 昆明医科大学学报

GCC/PKG/VASP,PKA/VASP et.al are associated with the occurrence and development of colorectal cancer.As one of the important signaling molecules of above signal pathways,VASP has constributed to the invasion,metastasis,apoptosis,angiogenesis and other events of colorectal cancer.The role of VASP signaling pathways in the development of CRC and its potential clinlic significance are reviewed in this article.

Effect of Cordycepin-Enriched WIB801C from Cordyceps militaris Suppressing Fibrinogen Binding to Glycoprotein IIb/IIIa

In this study, we investigated the effects of cordycepin-enriched (CE)-WIB801C, a n-butanol extract of Cordyceps militaris-hypha on collagen-stimulated platelet aggregation. CE-WIB801C dose dependently inhibited collagen-induced platelet aggregation, and had a synergistic effect together with cordycepin (W-cordycepin) from CE-WIB801C on the inhibition of collagen-induced platelet aggregation. CE-WIB801C and cordycepin stimulated the phosphorylation of VASP (Ser157) and the dephosphorylation of PI3K and Akt, and inhibited the binding of fibrinogen to glycoprotein IIb/IIIa (alphaIIb/beta3) and the release of ATP and serotonin in collagen-induced platelet aggregation. A-kinase inhibitor Rp-8-Br-cAMPS reduced CE-WIB801C-, and cordycepin-increased VASP (Ser157) phosphorylation, and increased CE-WIB801C-, and cordycepin-inhibited the fibrinogen binding to alphaIIb/beta3. Therefore, we demonstrate that CE-WIB801C-, and cordycepin-inhibited fibrinogen binding to alphaIIb/beta3 are due to stimulation of cAMP-dependent phosphorylation of VASP (Ser157), and inhibition of PI3K/Akt phosphorylation. These results strongly indicate that CE-WIB801C and cordycepin may have preventive or therapeutic potential for platelet aggregation-mediated diseases, such as thrombosis, myocardial infarction, atherosclerosis, and ischemic cerebrovascular disease.

Estudo da expressão e participação de VASP e Zyxin na diferenciação hematopoiética, na leucemia mieloide crônica e na via de sinalização BCR-ABL / VASP and Zyxin expression and participation in hematopoietic differentiation, in chronic myeloid leukemia and BCR-ABL signaling pathway

VASP (Vasodilator-stimulated phosphoprotein) e Zyxin são proteínas reguladoras de actina que controlam a adesão célula-célula. Zyxin dirige a montagem da actina através da interação e recrutamento da VASP a sítios específicos da adesão. A fosforilação da VASP ou da Zyxin altera suas atividades nas junções aderentes. PKA fosforila VASP em serina 157, regulando, assim, importantes funções celulares de VASP. VASP interage com ABL e é substrato da oncoproteína BCR-ABL. A presença da proteína BCR-ABL promove a oncogênese em pacientes com leucemia mieloide crônica (LMC) devido à ativação constitutiva da atividade tirosina quinase. Apesar de já descrita alteração da expressão de VASP e Zyxin em diferentes tumores epiteliais, o papel de VASP e Zyxin na LMC, na via de sinalização BCR-ABL e a participação destas proteínas na hematopoiese são desconhecidos. Desta maneira, demonstramos aqui ausência de p-VASP ser157 em células de medula óssea de pacientes com LMC, em contraste com a presença de p-VASP ser157 em doadores saudáveis. Pacientes com LMC em remissão, responsivos a inibidores de tirosina quinase, apresentam p-VASP ser157, enquanto os pacientes resistentes não expressam p-VASP ser157. Utilizando células K562 inibidas para VASP ou Zyxin, observamos que VASP e Zyxin modulam as proteínas anti-apoptóticas BCL-2 e BCL-XL da via de sinalização do BCR-ABL. Em adição, células K562 silenciadas para a VASP apresentam diminuição na atividade de FAK y925 e demonstramos que VASP interage com FAK. A expressão de VASP e Zyxin e de suas formas ativas aumenta durante a diferenciação megacariocítica e a inibição de VASP implica em diminuição na expressão do marcador CD61. Identificamos no presente estudo a participação de VASP e Zyxin na via do BCR-ABL, regulando a expressão de proteínas efetoras anti-apoptóticas e, também, na diferenciação megacariocítica...

VASP (vasodilator-stimulated phosphoprotein) and Zyxin are actin regulatory proteins that control cell-cell adhesion. Zyxin directs actin assembly by interacting and recruiting VASP to specific sites of adhesion. The phosphorylation of VASP and Zyxin modifies their activity in cell-cell junctions. PKA phosphorylates VASP at serine 157 regulating VASP cellular functions. VASP interacts with ABL and VASP is a substrate of BCR-ABL oncoprotein. The presence of BCR-ABL protein drives oncogenesis in patients with chronic myeloid leukemia (CML) due to a constitutive activation of tyrosine kinase activity. It has been described an altered expression of VASP and Zyxin in different types of tumor; however the function of VASP and Zyxin in CML, in BCR-ABL pathway and in hematopoiesis remains unknown. We describe here the absence of p-VASP ser157 in CML bone marrow cells, in contrast to p-VASP ser157 expression in healthy donors. Patients responsive to tyrosine kinase inhibitors present p-VASP ser157, while resistant patients do not have p-VASP ser157. In K562 cells we observed that VASP and Zyxin modulate anti-apoptotic proteins BCL-2 and BCL-XL. VASP depletion in K562 cells decreases FAK y925 activity and VASP interacts with FAK. Expression of VASP, p-VASP, Zyxin and p-Zyxin increases during megakaryocyte differentiation and VASP inhibition affects this differentiation through reduced CD61 expression in VASP depleted cells. We identify here the participation of VASP and Zyxin in BCR-ABL pathway affecting anti-apoptotic proteins and, also, in megakaryocyte differentiation. Then, the altered expression of VASP activity in CML patients may contribute to CML pathogenesis, affecting cellular differentiation or leukemic cell adhesion...

A randomised controlled trial of exercise and hot water bottle in the management of dysmenorrhoea in school girls of Chandigarh, India.

Objectives : To estimate the prevalence of primary dysmenorrhoea among school girls and to compare the impact of exercise and hot water bottle on the occurrence and severity of primary dysmenorrhoea among the study population. Material and methods : A cross sectional study was done to estimate the prevalence of dysmenorrhoea in two schools of Chandigarh, India. For the Randomised Controlled Trial, group randomisation of the two schools was done into 2 intervention groups (exercise & hot water bottle groups). 53 girls in school 1 and 75 girls in school 2 participated in the intervention. Comparison of baseline Menstrual Distress Questionnaire (MDQ) scores & Visual Analogue Scale for Pain (VASP) scores were done with 1st, 2nd & 3rd month post intervention scores using mean, standard deviation, t-test. Results : Prevalence of dysmenorrhoea was 60.7%. Median age of the school girls was 14 years. The mean VASP score decreased from 5.75 to 2.96 (P<0.0001) and from 5.16 to 2.06 (P<0.0001) at 3 months, in the exercise and hot water bottle group respectively. The mean MDQ score decreased from 14.53 to 7.85 (P<0.0001) and from 14.92 to 8.16 (P<0.0001) at 3 months, in the exercise and hot water bottle group respectively. Conclusion : Both exercise & hot water bottle can be used in dysmenorrhoeic girls in home setting to provide relief from pain and menstrual distress.

Effect of cell adhesion on vasodilator-stimulated phosphoprotein phosphorylation / 基础医学与临床

Objective To study the changes of Vasodilator-stimulated Phosphoprotein phosphorylation which was dynamically regulated by cell adhesion to extracellular matrix,and to investigate the roles of VASP in the process of tumor cell adhesion and de-adhesion.Methods HeLa cells were divided into four groups: stably adherent cells,cells held in suspension,cells replated onto dishes coated with collagen I and positive control(cells treated with Forskolin).Changes in cell morphology were observed in inverted phase contrast microscope.Phosphorylation of VASP was detected by Western blot,and PKA activity was measured by PKA assay kit.(Results In) untreated,stably adherent cells,VASP was predominantly in the unphosphorylated form; Detachment of cells stimulated PKA activity and induced PKA-dependent phosphorylation of VASP;VASP was phosphorylated gradually following reattachment but returned to base level of phosphorylation during active cell spreading.Conclusion During the process of HeLa cell adhesion and de-adhesion,dynamical changes of VASP from the form of phosphorylation to dephosphorylation via PKA have been observed.

Fetal vesico-amniotic shunting procedure (VASP) for posterior urethral valve syndrome at 19weeks' gestation / 대한산부인과학회잡지

An ultrasonographic examination revealed increased fetal bladder size and decreased AFI as well as fetal bilateral hydronephrosis at 173weeks' gestation. Diagnosis of the fetal posterior urethral valve syndrome was made. Percutaneous fetal bladder puncture with aspiration and amniocentesis was performed. The fetus was normal male karyotype and with a predicted good renal function(sodium concentration, chloride concentration, and osmolarity at 74 mEq/L, 60 mEq/L, and 148 mOsm, respectively). So, the fetus underwent amnioinfusion and vesico-amniotic shunting procedure (VASP) using a double-basket catheter at 194weeks' gestation in order to prevent development of dysplastic kidneys and hypoplastic lungs. The healthy male baby was delivered at 384weeks' gestation and had normally functioning kidney. Cutaneous vesicostomy was performed for the newborn since the urethral orifice was small. The one year old infant is now well and waiting for urethroscopic valve ablation procedure.