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Objective:To investigate the expression of platelet receptor CD62P in septic rats and the anti-inflammatory effect of ticagrelor and its protective effect on myocardial injury in septic rats.Methods:Thirty-two male SD rats were randomly(random number) divided into 4 groups: sham group, cecal ligation and puncture group(CLP), low dose group: the dose of 10 mg/kg, high dose group: the dose was 50 mg/kg, 8 rats in each group. The rats in the sham operation group were only treated with abdominal switch and cecum stripping, and the rats in the sepsis group, the low dose group and the high dose group were treated with CLP method to establish the sepsis model. The rats in the ticagrelor administration group were treated with ticagrelor at a dose of 10 mg/kg and 50mg/kg by gavage, respectively. The sham operation group and the sepsis group were treated with normal saline (1 mL/kg) by gavage. The rats were administrated twice by gavage 12 hours before and 12 hours after surgery. Blood samples were collected from the abdominal aorta 24 hours after modeling and then pathological specimens were collected. The expression of platelet surface receptor CD62P was detected by flow cytometry. The levels of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were detected by enzyme-linked immunosorbent assay (ELISA). The levels of myocardial injury markers including CKMB and LDH were detected. The levels of transaminase, creatinine and white blood cell were detected. HE staining was used to observe the pathological morphology of myocardial tissue. Cardiomyocyte apoptosis was observed by TUNEL assay.Results:① Compared with sham group, the expression of CD62P in CLP group significantly increased ( P<0.01). Compared with the CLP group, the expression levels of CD62P in the two treatment groups significantly decreased, and the HD group was more significant ( P <0.01).②ELISA results showed that compared with sham group, the level of IL-6 in CLP group was significantly increased ( P<0.05). Compared with the CLP group, the HD group significantly decreased ( P< 0.05). There was no significant decrease in IL-6 level in the LD group. The level of TNF-α in CLP group was significantly higher than that in sham group ( P< 0.01). ③ Compared with sham group, the expression levels of CKMB and LDH in CLP group and two ticagrelor intervention groups significantly increased ( P <0.01). Compared with the CLP group, CKMB and LDH in the treatment group significantly decreased ( P <0.05), and the HD group decreased more significantly ( P<0.01). ④ Compared with sham group, WBC, ALT, CR values in CLP group significantly increased, while after the intervention with ticagrelor, WBC, ALT, CR values in rats significantly decreased ( P <0.05), and the difference significantly related to the dose. ⑤ The pathological results showed that the morphology of myocardial cells in sham group was normal. The CLP group most myocardial cell injury. LD and HD group the CLP group obviously reduce myocardial cell injury.⑥ Tunel staining showed that compared with a small number of positive cells in Sham group, a large number of positive cells were stained in CLP group. The apoptosis of myocardial cells in LD and HD groups significantly reduced compared with CLP group. Conclusions:Sepsis activates platelets and stimulates the overexpression of CD62P, which induces excessive activation of inflammatory response, induces apoptosis and damage of cardiomyocytes, and leads to septic myocardial injury. The cardioprotective effect of ticagrelor may be related to the inhibition of the reduction of CD62p expression after platelet activation, and the expression level of CD62p has a dose-dependent relationship with ticagrelor.
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【Objective】 To investigate the effect of trehalose added to M-Sol on platelet preservation at 4℃ in vitro. 【Methods】 Seven bags of ABO homotype concentrated platelets were randomly selected and divided into 7 groups according to storage temperature, preservation medium and trehalose concentration: 22℃+ PRP group, PRP group, M-Sol+ PRP group, M-Sol+ PRP+ 1 g/L trehalose group, M-Sol+ PRP+ 5 g/L trehalose group, M-Sol+ PRP+ 15 g/L trehalose group and M-Sol+ PRP+ 25 g/L trehalose group. PRP group and M-Sol preservation groups were stored at 4℃. Plt, PDW, MPV, maximum aggregation, CD62p positive rate, GLU and LAC concentration were detected on the 1st, 3rd, 5th and 7th day after preservation, and the changes of GLU and LAC concentration within 7 days were calculated. 【Results】 With the extension of preservation time, Plt decreased in all groups, and there was no significant difference among groups at the same time (P>0.05). PDW and MPV increased in all groups. When preserved to the 7th day, the PDW 10.22±0.43(fL) and MPV 8.74±0.40(fL) of M-Sol+ PRP+ 5 g/L trehalose group were the lowest, which was significantly different from that of 22℃+ PRP group (P0.05). During the preservation period, the maximum aggregation degree of each group decreased gradually. Except for the PRP group, the maximum aggregation degree of the M-Sol+ PRP+ 5g/L trehalose group was the highest(27.29±6.62), which was significantly higher than that of the 22℃+ PRP group on the 7th day after preservation (P<0.05). On the 7th day after preservation, the positive rate of CD62p in M-Sol+ PRP+ 5g/L trehalose group was the lowest(15.46±2.46), and there was significantly different compared with other groups (P<0.05). 【Conclusion】 Adding appropriate concentration of trehalose to M-Sol can inhibit platelet activation at 4℃ and reduce platelet storage lesion.
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To observe the contribution of FoxO1 on its downstream molecules expression and function in Jurkat cells,Jurkat cells were infected with FoxO1 expression lentivirus and interference lentivirus to establish FoxO1-KLF2-S1P1 signaling pathways mod-el.After infection of 120 h,FoxO1,KLF2,S1P1 and CD62L mRNA levels and the protein expression of FoxO1,FoxO1-p and KLF2 were increased (P<0.05) in FoxO1-overexpression group.Converse results(P<0.05) were observed in the interference group.The proportions of S1P1+cells were increased in both groups.It was notably that S1P1+cells were decreased(P<0.05) in interference group after infection of 72 h.The proportion of CD62L+cells was increased(P<0.05) in overexpression group,it was decreased(P<0.05) in the interference group.This vitro experiments showed S1P1 and CD62L could be regulated by FoxO1 lentivirus,and provided an experimental basis for research about intracellular FoxO1-KLF2-S1P1 signaling pathways and cellular functions.
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Objective To investigate the effects of endothelial cell, neutrophil and platelet activation on the development and thrombophilia of malignant lymphoma. Methods The levels of cell activating factors soluble thrombomodulin (sTM) in 40 patients with malignant lymphoma and 30 healthy controls in Jiangdu People ''s Hospital of Yangzhou from October 2009 to November 2016 were tested by enzyme linked immunoabsorbent assay (ELISA). The flow cytometry (FCM) method was used to measure the level of CD11b on the surface of anti-freezing whole blood neutrophil and CD62p on the surface of platelet. The results were analyzed by ANOVA and LSD-t test. Results The level of sTM in patients with malignant lymphoma before treatment was significantly higher than that after treatment and in the control group [(49.7±9.3) ng/ml vs. (6.3± 1.8) ng/ml, (5.6±1.5) ng/ml respectively;F=736.633, P=0.000]. There were significant differences between the levels before and after treatment and before treatment and in the control group (t= 33.789, P= 0.000;t=31.782, P=0.000). The average fluorescence intensity (MFI) of neutrophil surface CD11b in patients with malignant lymphoma before treatment was significantly higher than that after treatment and in the control group (184 ± 43 vs. 118 ± 12, 112 ± 15 respectively; F=101.845, P=0.000). There were significant differences between the levels before and after treatment and before treatment and in the control group (t=12.228, P= 0.000; t= 12.184, P= 0.000). The level of platelet surface CD62p in patients with malignant lymphoma before treatment was significantly higher than that after treatment and in the control group [(6.4 ± 2.4) ％ vs. (1.2 ± 0.7) ％, (1.3 ± 0.5) ％ respectively; F= 141.481, P= 0.000]. There were significant differences between the levels before and after treatment and before treatment and in the control group (t=15.010, P= 0.000; t= 13.679, P= 0.000). Conclusions The levels of cell activating factors might be correlated with the progression of malignant lymphoma. Furthermore, endothelial cell, neutrophil and platelet activation have accelerated the development of malignant lymphoma, and the interaction among them may result in the prethrombotic state.
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Objective To explore the correlation between CD62p and clopidogrel resistance in patients undergoing percutaneous coronary intervention with coronary heart disease combined with diabetes mellitus.Methods 150 patients ndergoing percutaneous coronary intervention with coronary heart disease combined with diabetes mellitus were enrolled in our hospital from August 2013 to December 2016,of all patients accepted clopidogrel treatment,the platelet aggregation rate were analyzed before and after treatment,which all patients divided into two groups as the range of decreasing platelet aggregation rate whether ≥ 10％,clopidogrel effective group (C2) and clopidogrel resistance group (C 1) were detected CD62p and platelet aggregation maximum (PAGM),the relevance of which range with platelet aggregation rate was analyzed.Results After treatment,the platelet aggregation rate of all patient decreased signifieantly (P ＜ 0.05).Of all patients divided into clopidogrel effective group (C2) and clopidogrel resistance group (C 1) as the range of decreasing platelet aggregation rate,and the CD62p of Group C1 decreased compared with before treatment (P ＜ 0.05),the level of CD62p and PAGM in Group C2 were decreasing compared with pre-treatment,and the range of which was larger than those Group C1 (P ＜ 0.05).The reference of CD62p and PAGM in Group C2 (r =0.424,P =0.001) was better than that Group C1 (r =0.387,P =0.020).Conclusions Clopidogrel for patients undergoing percutaneous coronary intervention with coronary heart disease combined with diabetes mellitus possessed inhibition for platelet activity,and clopidogrel resistance might be associated with platelet activity.
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Objective To explore the relationship between platelet activating factors and four platelet parameters in diabetic retinopathy (DR).Methods Eighty patients in our hospital were chosen from January 2015 to December 2015,regular ophthalmological examination and fundus fluorescence were performed on all cases.On the basis of international clinical diabetic retinopathy classification standard,the patients were divided into four groups:No DR group with 20 cases,non-proliferative diabetic retinopathy (NPDR) group with 20 cases,and proliferative diabetic retinopathy (PDR) group with 20 cases.20 healthy people who received physical examination during the same period were recruited as the control group.The flow cytometry instrument was used to determinate platelet activating factor CD62p and CD63,automatic blood analyzer was used to determinate platelet parameter which including blood platelet count (PLT),mean platelet volume (MPV),plateletcrit (PCT) and platelet distribution width (PDW),and the changes of platelet activation factor,parameters and its clinical significance in DR were discussed.Results With the development of DR,CD62p,CD63,MVP were numerical gradually increased,and PLT were numerical gradually reduced,the differences were statistically significant (all P ＜ 0.05);Compared with control group,CD62p,CD63,MVP were increased and PLT were reduced in other group,the differences were statistically significant (all P ＜ 0.05);Compared with pure diabetes group,CD62p,CD63,MVP were increased and PLT were reduced in NPDR group and PDR group,the differences were statistically significant (all P ＜ 0.05);Compared with NPDR group,CD62p,CD63,MVP were increased and PLT were reduced in PDR group,the differences were statistically significant(all P ＜ 0.05).PDW were numerical gradually increased in control group,pure diabetes group,NPDR group,PDR group,the differences were statistically significant(all P ＜ 0.05);Control group compared with pure diabetes group,NPDR group compared with PDR group,the differences in PDW was not statistical significant (all P ＞ 0.05);But compared with control group or pure diabetes group,PDW of NPDR group and PDR group increased,the differences were statistically significant(all P ＜ 0.05).The differences in PCT among groups was not statistically significant(all P ＞ 0.05).Conclusion The platelet activating factor CD62p,CD63 and parameters of platelet show differences in each diabetic retinopathy period,platelet activating factor in the occurrence and development of diabetic retinopathy have important significance.Monitor the platelet activating factor and parameters of platelet can help surveillance of the disease.
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Objective To adopt 5 kinds of platelets aggregation function analyzer to explore and study the reliable method for detecting the platelets aggregation function and accurate parameters .Methods The platelets aggregation function in the control group and the single clopidogrel group was simultaneously detected by utilizing the ADP induced light turbidimetric platelet aggre‐gation analyzer (LTA) test ,flow cytometry for PAC‐1(receptor of Fg ,Ca2+ ,GPⅡb/Ⅲa forming complex) and CD62p(P selectin) activation percentage detection test ,Innova PL‐11 for platelets analysis test ,VerifyNow anti‐platelet therapy monitoring system and thrombelastogram(TEG) .Results (1)The 6‐parameter differences of ADP% ,PAC‐1 and CD62p receptor activation percentage ac‐tivated by ADP ,MAR% ,INHI% ,ADP induced MA value(mm) detected by TEG had statistical differences between the control group and the case group(P0 .05) .(2)ADP% was positively correlated with (100‐INHI)% ,MAR% ,ADP activated CD62p and PAC‐1 receptor activation percentage(r=0 .565 ,0 .939 ,0 .769 ,0 .583 ,P<0 .05 );while which had no correlation with ADP induced platelets aggregation value(% Agg) detected by TEG(r=0 .794 ,0 .715 ,0 .889) .Conclusion (1)Clopidogre has the anti‐platelets effect .(2)LTA is easily operating ,cheap and stable in detection results ,has high popularizing rate in hospitals and is the first option method for clinically monitoring the platelets aggregation function .
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Objective To understand how Vibrio vulnificus hemolysin (VvhA) affects the viability of murine liver CD4+ T cells as well as its effects on the numbers of mitochondria and the expression of CD62L. Methods The primary murine liver monocytes (MNs) were isolated from C57BL/ 6 mice and then treated with recombinant VvhA (rVvhA) for 6 hours in vitro. The viability of murine liver CD4+T cells and the expression of CD62L were measured by staining with anti-mouse CD4, CD8, CD44, CD62L and cell via-bility fluorescent dye or fluorescent antibody. Moreover, the cells were simply incubated with MitoTracker or JC-1 probes to label mitochondria and mitochondrial membrane potential, which were further analyzed by using flow cytometry analysis. Results With the increase in the doses of rVvhA, the viability of murine liv-er CD4+T cells was decreased from 81. 5% to 15. 8% . The expression of CD62L on the surface of murine liver CD4+T cells was dramatically decreased. Both the murine liver na?ve and effector CD4+ T cells were sensitive to the cytotoxicity of rVvhA. Moreover, treating murine liver CD4+ T cells with rVvhA resulted in significantly decreased numbers of mitochondria and lower mitochondrial membrane potential. Conclusion The cytotoxicity of rVvhA to murine liver CD4+T cells might be achieved through inhibiting the expression of CD62L, decreasing the numbers of mitochondria and lowering mitochondrial membrane potential.
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High levels of low-density lipoprotein cholesterol (LDL-C) enhance platelet activation, whereas high levels of high-density lipoprotein cholesterol (HDL-C) exert a cardioprotective effect. However, the effects on platelet activation of high levels of LDL-C combined with low levels of HDL-C (HLC) have not yet been reported. We aimed to evaluate the platelet activation marker of HLC patients and investigate the antiplatelet effect of atorvastatin on this population. Forty-eight patients with high levels of LDL-C were enrolled. Among these, 23 had HLC and the other 25 had high levels of LDL-C combined with normal levels of HDL-C (HNC). A total of 35 normocholesterolemic (NOMC) volunteers were included as controls. Whole blood flow cytometry and platelet aggregation measurements were performed on all participants to detect the following platelet activation markers: CD62p (P-selectin), PAC-1 (GPIIb/IIIa), and maximal platelet aggregation (MPAG). A daily dose of 20 mg atorvastatin was administered to patients with high levels of LDL-C, and the above assessments were obtained at baseline and after 1 and 2 months of treatment. The expression of platelets CD62p and PAC-1 was increased in HNC patients compared to NOMC volunteers (P<0.01 and P<0.05). Furthermore, the surface expression of platelets CD62p and PAC-1 was greater among HLC patients than among HNC patients (P<0.01 and P<0.05). Although the expression of CD62p and PAC-1 decreased significantly after atorvastatin treatment, it remained higher in the HLC group than in the HNC group (P<0.05 and P=0.116). The reduction of HDL-C further increased platelet activation in patients with high levels of LDL-C. Platelet activation remained higher among HLC patients regardless of atorvastatin treatment.
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Adolescente , Niño , Femenino , Humanos , Masculino , Logro , Trastorno por Déficit de Atención con Hiperactividad/psicología , Atención/fisiología , Análisis de Varianza , Trastorno por Déficit de Atención con Hiperactividad/diagnóstico , Estudios de Cohortes , Escolaridad , Escalas de Valoración Psiquiátrica , Sensibilidad y EspecificidadRESUMEN
Objective To detect clopidogrel effect with light transmission aggregometry (LTA)and flow cytometry (FC). Methods ①Venous blood samples were taken from 71 inpatient with acute corotary syndrome (ACS)in PLA General Hos-pital,including unstable anqina,ST segment elevation myocardial infarction and non ST segment elevation myocardial infarc-tion (46 males,25 females)by random number table from June 2011 to March 2012,whose average age was 69(57~92).②All of them were served 160 mg aspirin and 300 mg clopidogrel after they were in hospital in the beginning,and then served with 75 mg/d clopidogrel for 6 months.On some day,firstly,they were required withdrawing drug for 10 days,and then ve-nous blood samples were separately taken from them before their served-clopidogrel again and their severd-clopidogrel 2 hours later.③The samples were assayed with LTA and FC simultaneously and the platelet aggregation rates before served-clopidogrel (ADPLTA-before serving ),platelet aggregation rates after served-clopidogrel (ADPLTA-after serving ),inhibition rates (ADPLTA-INDU ),PAC-1 activity percentage before served-clopidogrel (PAC-1 before serving ),PAC-1 activity percentage after served-clopidogrel (PAC-1 after serving ),inhibition rates (PAC-1 INHI ),CD62p activity percentage before served-clopidogrel (CD62pbefore serving ),CD62pactivity percentage after served-clopidogrel (CD62pafter serving ),inhibition rates (CD62pINHI )weregotten.All volunteers were signed informed consents and the experiment was approved by the hospital ethics committee.Re-sults ①The paired samples t-test was (t=-2.082,P =0.041)between ADPLTA-before serving (0%~97%)and ADPLTA-after serving (12%~97%),the paired samples t-test was (t = 3.663,P < 0.01)between PAC-1 before serving (15.1% ~ 78.9%)and PAC-1 after serving (14.5% ~ 78.3%);the paired samples t-test was (t = 2.082 and P = 0.041)between CD62pbefore serving (1.5% ~80.8%)and CD62pafter serving (1.4%~41.4%).②The pearson coeffcient correlation results were:ADPLTA-INDU (0%~28.2%) and PAC-1 INHI (0.6%~ 9.1%)(r = 0.297,P = 0.012);ADPLTA-INDU (0% ~ 28.2%)and CD62pINHI (0.1% ~ 48.5%)(r =0.220,P =0.065);PAC-1 INHI (0.6%~9.1%)and CD62pINHI (0.1%~48.5%)(r=0.736,P <0.001).Conclusion Because the correlation was bad between the inhibition rates of clopidogrel detected by FC and them by LTA,FC didn’t apply to clin-ical routine examination of the platelet aggregation.But it could be used to scientific researchs and auxiliary confirmation of routine examination results.
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Objective To explore the level of CD62E+ endothelial microparticles in acute cerebral infarction patients and its relationship with vascular risk factors,degree of severity and prognosis.Methods Seventy patients with acute cerebral infarction and 70 non-cerebral infarction patients (controls),admitted to our hospital from January 2013 to September 2013,were selected in our study; U.S.national institutes of health stroke scale (NIHSS) was performed to evaluate the degree of severity; Barthel index (BI) was used to evaluate the prognosis; the levels of blood lipids,blood glucose,uric acid,fibrinogen (FIB),homocysteine (Hcy),C-reactive protein (CRP) and lipoprotein-associated phospholipase A2 (Lp-PLA2) were routinely measured; CD62E+ levels were detected by flow cytometry,and the relationships of CD62E+ level with vascular risk factors,degree of disease and prognosis were analyzed.Results The blood CD62E+ level of patients with acute cerebral infarction ([1.55 ±0.67]microparticle/μL) was significantly higher than that in the control group ([1.01 ±0.66] microparticle/μL,P＜0.05); CD62E+ level in patients with NIHSS scores＞5 ([2.03±0.61] microparticle/μL) was significantly higher than that in patients with NIHSS scores ≤ 5 ([1.25±0.37] microparticle/μL,P＜0.05).Spearman correlation analysis showed that CD62E+ level was significantly correlated with the stroke severity (NHISS scores,r=0.537,P=0.000) and lipoprotein-associated phospholipase A2 (Lp-PLA2) level (r=0.327,P=0.006),and negatively correlated with prognosis indicator (r=-0.634,P=0.000); other factors showed no significant correlation.Conclusion CD62E+ microparticle can reflect the severity and prognosis of acute brain infarction,and it may be an important biological marker for clinical application.Besides,the correlation between Lp-PLA2 and CD62E+ means that the level ofCD62E+ micro particles is associated with the stability of athemsclerotic plaques.
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Objective To investigate the expression and clinical significance of platelet glycoprotein CD62p,CD63 and neutrophil surface CD64 in sepsis.Methods Fifty-six children with sepsis from March 2010 to March 2013 in Communicable Disease Department of our hospital were divided into severe sepsis group(n =16) and general sepsis group (n =40),normal control group included 34 subjects from health check.CD62p,CD63 and CD64 were detected by flow cytometry in children with sepsis,and compared with normal control group.Results The levels of CD62p,CD63 and CD64 in severe sepsis group were higher than those of general sepsis group (P ＜ 0.01).The levels of CD62p,CD63 and CD64 in general sepsis group were higher than those of normal control group (P ＜ 0.01).Correlation analysis indicated that CD62p and CD63 were in positive correlation with CD64 in children with sepsis(r =0.817,0.796,P ＜0.001).The positive correlations of CD62p,CD63 and CD64 with pediatric critical illness score were also found(CD62p:r =0.883,P ＜0.001;CD63:r=0.862,P ＜0.001;CD64:r=0.805,P ＜0.001).Conclusion CD62p,CD63 and CD64 are closely related to the severity of infection and diseases,and may be used as immune parameters for the estimation of the clinical severity and the prognosis of acute and severe diseases.
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Objective To investigate the change of serum CD62p of coronary heart disease (CHD) patients after different doses of clopidogrel administration.Methods One hundred and ninety-one patients with CHD were selected as our subjects.Of which,95 cases were with SAP and 66 cases were with non ST segment elevation acute coronary syndrome (NST-ACS).SAP patients were randomly given clopidogrel at dose of 75 mg/d or 150 mg/d and served as A and B groups.NST-ACS patients were randomly given 300 mg clopidogrel,then randomly divided into C and D groups with sequentially taking clopidogrel at dose of 75 mg/d or 150 mg/d respectively.Thirty healthy peoples were served as E group without drug intervention.Concentrations of serum CD62p were detected by Elisa before taking clopidogrel,24 h and the fifty day of after taking clopidogrel.Results (1) Before taking clopidogrel,the serum concentrations of CD62p in CHD patients were higher (A group:(7.62 ± 2.99) ng/L,B group:(8.48 ± 3.13) ng/L,C group:(9.50 ± 3.32) ng/L,D group:(10.22 ± 5.14) ng/L than that of healthy control group ((5.49 ± 1.99) ng/L,P ＜ 0.05).The Serum CD62p levels in SAP patients were lower than that of NST-ACS patients (P ＜ 0.05).(2)The serum concentrations of CD62p in A and B groups at before taking clopidogrel were (7.62 ±2.99) ng/L and (8.48 ±3.13) ng/L respectively,higher than that four days after taking clopidogrel ((6.79 ± 2.51) ng/L,(6.37 ± 1.80) ng/L;t =2.390,4.520;P ＜0.05 or P ＜0.01).There was no statistical significant difference between A and B groups(P ＞0.05).(3) In C and D groups,the serum CD62p at before taking clopidogrel were (9.50 ±3.32) ng/L and (10.22 ±5.14) ng/L,higher than that after taking clopidogrel for four days ((8.21 ± 2.62) ng/L,(8.17 ± 2.37) ng/L; t =2.084,2.157 ; P ＜ 0.05).No significant difference was seen between C and D groups (P ＞ 0.05).Conclusion The serum CD62p in patients with CHD was higher than that in the healthy control.Clopidogrel administration can decrease serum CD62p in CHD patients.
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Objective To investigate the changes of platelet aggregation rate and platelet surface CD 41+CD62P+ expression af-ter clopidogrel discontinuation in the patients with percutaneous coronary interventions (PCI)) .Methods The platelet aggregation rates and platelet surface CD41+CD62P+ in 52 PCI patients with oral clopidogrel for near 12 months and discontinuation soon were measured before clopidogrel therapy(T0 ) ,in 1 week after clopidogrel therapy(T1 ) ,1 week before clopidogrel discontinuation(T2 ) , 1 week(T3 ) and 1 month after clopidogrel discontinuation (T4 ) .Results Compared with T0 ,the platelet aggregation rate and the expression of platelet CD41+CD62P+ at T1 were significantly decreased ,the difference showing statistical significance (P<0 .05) , which at T2 maintained the lower levels ;which at T3 were increased ,which at T4 were recovered to those at T0 .The platelet aggre-gation rates at various time points were (44 .20 ± 18 .36)% ,(25 .38 ± 12 .10)% ,(23 .74 ± 8 .15)% ,(51 .79 ± 10 .55)% and(45 .97 ± 16 .42)% respectively ,and the positive rates of CD41+ CD62P+ were(12 .96 ± 11 .48)% ,(3 .93 ± 3 .33)% ,(4 .72 ± 3 .14)% , (13 .90 ± 10 .38)% and(10 .84 ± 8 .13)% ,respectively .Conclusion In the patients treated with 12-month clopidogrel after PCI ,the platelet aggregation rate and the CD41+CD62P+ positive rate are mildly increased at 1 week after clopidogrel discontinuation and gradually returned to the level before discontinuation at 1 month after clopidogrel discontinuation .
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Objective To investigate the clinical significance of platelet activation markers CD62p and PAC-1 in patients with hypertension and coronary heart diease.Methods To select patients who were in the Second Affiliated Hospital of Dalian Medical University between January 2012 and September 2012,42 patients with hypertension,46 patients with coronary heart disease.Flow cytometry was used to test the positive percentage of the platelet surface activation markers CD62p and PAC-1 .To observe the difference among hypertension group,coronary heart disease group and health control group;hyper-tension and coronary heart disease group.Results The positive percentage of platelet activation markers CD62p and PAC-1 in hypertension (36.36±9.62,7.18±8.20)%,coronary heart disease group (42.74±14.60,8.81±12.53)% showed sta-tistically significant differences (t=4.150~5.853,P<0.01)compared with healthy control group (26.82±9.13 ,1.09± 1.05)%.The positive percentage of CD62p in coronary heart disease group (42.74±14.60)% was also higher than that of hypertension (36.36±9.62)%,and the difference between them was of statistical significance (t=2.444,P<0.05).Con-clusion The hypertensive patients and coronary heart disease were in pre-thrombotic state,the activation of platelet increase significantly.Molecular markers should be measured in hypertensive patients and coronary heart disease for prevention and treatment of thrombotic disease complication.
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Objective To observe the clinical findings about the endothelial cell injury related to the genesis of inflammatory cytokines and coagulation.Methods A total of 70 critically ill patients with SIRS (systemic inflammatory response syndrome) admitted to intensive care unit (ICU) between September 2009 and February 2010 were enrolled for a prospective and control study.According to diagnostic criteria of Sepsis/SIRS,the patients were divided into two groups:sepsis group (n =38) and SIRS group (n =32),and another 20 healthy volunteers served as control group.Patients in the sepsis group and SIRS group were matched by clinical signs of high blood pressure,diabetes and its complications.The demographics of patients including age,sex,body mass index (BMI),smoking and alcohol addict were comparable among the different groups.The 6 ml peripheral blood samples were collected within 24 h after admission to ICU for enzyme-linked immunosorbent assay (ELISA) to detect the plasma levels of s-CD62P,TNF-α,and hsCRP.And variables of coagulation function such as platelet (PLT),prothrombin time (PT),activated partial thromboplastin time (APTT),D-dimer and antithrombin-Ⅲ (AT-Ⅲ) were analyzed during 24 h after admission to ICU.Meanwhile sequential organ failure assessment (SOFA) score of critically ill patients was evaluated.Data were expressed in mean ± standard deviation and were statistically analyzed by using SPSS 17.0 statistical software.The differences in plasma levels of s-CD62P of patients in each group were analyzed by ANOVA and Kruskal Wallis test.The relationship between s-CD62P and inflammatory cytokines as well as with coagulation were determined by Pearson correlation analysis.Changes were considered as statistically significant if P value was less than 0.05.Results ① Compared with control group and SIRS group,the levels of s-CD62P,TNF-α and high sensitive C-reactive protein (hs-CRP) were significantly higher in sepsis group (P ＜ 0.05).② The plasma levels of D-dimer,PT,APTT in sepsis group and SIRS group were significantly higher than those in control group,while the platelet count (PLT) and the activity of AT-Ⅲ were obviously lower (P ＜ 0.05).③ In sepsis group,the plasma levels of hs-CRP and TNF-α positively correlated with PT,APTT,D-dimer,and negatively correlated with AT-Ⅲ,PLT (P ＜ 0.05).④ Plasma levels of s-CD62P were significantly correlated with plasma levels of TNF-α,hs-CRP,D-dimer,PT,APTT,whereas correlated negatively well with PLT,AT-Ⅲ (P ＜ 0.05).Conclusions The plasma s-CD62P concentration is elevated as a early biomarker in patients with sepsis,and it acted as one of pathogenic factors responsible for endothelial cell damage.Coagulation and mediators of inflammation promotes each other,aggravating the severity of the sepsis.The plasma s-CD62P may be the important factor associated with initiation of coagulation development and inflammatory reaction.
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@#BACKGROUND: Current studies on CD62P have focused mainly on cardiovascular diseases, while only few studies have evaluated the effects of CD62P on the development of sepsis and the association between endothelial cell injury with inflammation and coagulation. This study attended to explore the association between endothelial cell injury with inflammation and coagulation by evaluating the expression of soluble CD62P (s-CD62P) in plasma and its mechanism in patients with sepsis, thus to provide the evidence of effective treatment of sepsis with anti-adhesion therapy targeted CD62P. METHODS: A total of 70 critically ill patients with systemic inflammatory response syndrome (SIRS) admitted to intensive care unit (ICU) between September 2009 and February 2010 were enrol ed for a prospective and control study. According to the diagnostic criteria of sepsis/SIRS, the patients were divided into two groups: a sepsis group (n=38) and a SIRS group (n=32). Another 20 healthy volunteers served as a control group. Patients in the sepsis group and SIRS group were matched by clinical signs of high blood pressure, diabetes and its complications. The demographics of the patients including age, sex, body mass index (BMI), smoking and alcohol addict were compared among the groups. Six mL peripheral blood samples were collected within 24-hour admission in ICU for enzyme-linked immunosorbent assay (ELISA) to detect the plasma levels of s-CD62P, TNF-α, and hs-CRP. And variables of coagulation function such as platelet (PLT), prothrombin (PT), activated partial thromboplastin time (APTT), D-dimer and antithrombin-III (AT-III) were analyzed during 24 hours after admission to ICU. Meanwhile sequential organ failure assessment (SOFA) score of critically ill patients was evaluated. Data were expressed as mean±standard deviation and were statistical y analyzed by using SPSS 17.0 statistical software. The differences in plasma levels of s-CD62P of patients in each group were analyzed by ANOVA and the Kruskal-Wallis test. The relations between s-CD62P and inflammatory cytokines as well as with coagulation were determined by Pearson's product moment correlation coefficient analysis. Changes were considered as statistically significant if P value was less than 0.05. RESULTS: Compared with the control group and SIRS group, the sepsis group demonstrated significantly higher levels of s-CD62P, TNF-α and highly sensitive C-reactive protein (hs-CRP) (P<0.05). The plasma levels of D-dimer, PT, and APTT in the sepsis and SIRS groups were significantly higher than those in the control group, while the platelet count and the activity of AT-III were obviously lower (P<0.05). In the sepsis group, the plasma levels of hs-CRP and TNF-α were positively correlated with PT, APTT, and D-dimer, and negatively correlated with AT-III and PLT (P<0.05). The plasma levels of s-CD62P were significantly correlated with the plasma levels of TNF-α, hs-CRP, D-dimer, PT, and APTT, whereas they were correlated negatively well with PLT and AT-III (P<0.05). CONCLUSIONS: The concentration of plasma s-CD62P is elevated as a early biomarker in patients with sepsis, and it serves as one of the pathogenic factors responsible for endothelial cell damage. Coagulation and mediators of inflammation promote each other, aggravating the severity of sepsis. Plasma s-CD62P may be an important factor for the development of coagulation and inflammatory reaction.
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Platelet flow cytometry is an emerging tool in diagnostic and therapeutic hematology. It is eminently suited to study the expression of platelet surface receptors both qualitatively as well as quantitatively. It can serve as a useful marker for the documentation of in vivo platelet activation, and thus, fore-warn the risk of thromboembolism in patients with diabetes mellitus, coronary syndromes, peripheral vascular diseases, and pre-eclampsia. This technique can also be extended to study and compare the effect of various antiplatelet drugs on the level of activation of platelets and to establish any dose-effect relationship of these drugs. Topographical localization of platelet granules and study of platelet-platelet and platelet-leukocyte interaction is also possible by this procedure. All these parameters serve as pointers towards the presence of activated platelets in the circulation with its thromboembolic consequences. This is a simple reliable and cost effective technique which has a wide application in the diagnosis of various inherited and acquired platelet disorders. Study of platelet cluster of differentiation (CD) markers in various inherited disorders i.e. Bernard Soulier’s disease, von Willebrand disease, Glanzman’s disease, and Grey platelet syndrome may help categories the molecular lesions in these oft under-studied disorders.
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Objective To analyze the changes of PAIgG, CD62P, CD4+CD25+ Foxp3+Tr,and IL-18 before and after treatment in peripheral blood of children with acute idiopathic thrombocytopenic purpura(ITP) and investigate the function of these factors in the pathogenesis of ITP.Methods Forty-one cases of acute ITP children were divided into the effective group(35cases) and the ineffective group (6cases) according to the clinical treatment. To detect PAIgG,CD62P,and the number of Tr cells by using flow cytometry ,IL-18 plasma levels by ELISA assay,and analyze the variations of these indicators before and after treatment in children with acute ITP. Results In the effective treatment group, PAIgG, CD62P before treatment were 53.05%,(14.18±5.04 )%, which were significantly higher than that after treatment [18.62%, ( 8.36±1.95 )%] and control group[5.26%,(2.65±0.59) %,all P<0.01],and PAIgG,CD62P after treatment were also higher than that in control group [all P<0.05].IL-18,CD4 + T lymphocytes, Tr/CD4+T-lymphocyte ratios before treatment [415.47 ±38.92 ) ng/L,( 25.64 ± 5.81 )%,( 2.67 ± 0.14 )%]were significantly lower than that after treatment [(512.85±42. 17)ng/L,(35.08±6.07)% ,(4.76±0.58)%] and control group[(506. 39±32.28) ng/L,(35.32±2.27)% ,(5.37 ±0.69)% ,all P<0.01]. IL-18, CD4 +T lymphocytes, Tr/CD4 +T-lymphocyte ratios after treatmenthad no statistically significant difference compared with control group( all P<0.05 ). In ineffective group, the test results of PAIgG, CD62P, IL-18, CD4 +T lymphocytes, Tr/CD4+ T-lymphocyte ratios showed no significant change before and after treatment( all P<0.05 ).IL-18 had negative correlations with PAIgG,CD62P respectively before and after treatment(all P<0.05 ). Tr cells / CD4 + T had negative correlations with PAIgG,CD62P respectively (all P<0.05). Conclusions The amount of Tr, IL-18 were reduced, while CD62P and PAIgG increased in peripheral blood of children with acute ITP. IL-18, Tr , CD62P and PAIgG play important roles in the pathogenesis of acute ITP.
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Objective To investgate role of TLR2 in the activation, the innate immune and inflammation of human platelets. Methods Human washed platelets were separated from healthy people(n=5) and were stimulated with different concentrations(1μg/ml, 5μg/ml, 10μg/ml) of TLR2 agonistPam3CSK4(a synthetic bacterial lipoproteins). Then the platelet aggregation rate, the expression of CD62p and TLR2 on the platelet surface were measured. Results The platelet aggregation rate were (28.32±5.67)%, (52.56±8.54)% and (76.24±11.23)%, respectively, at concentration of 1μg/ml, 5μg/mland 10μg/ml of Pam3CSK4, more than (12.83±2.43)% at 0μg/ml of it. In addition, the expression of CD62p were (18.45±2.66)%, (22.45±2.04)%, (29.53±4.08)%, respectively at above concentration of Pam3 CSK4, more than (11.20±1.67)% of CD62p at control group(P<0.01). The expression of TLR2 was not significantly increased at a lower concentration of Pam3CSK4(1μg/ml) with (16.85±6.10)% compared with(10.81±3.99)% at the control group. However, it were (21.15±9.90)% and (22.52±9.26)%, respectively, at a higher concentration(5μg/ml, 10μg/ml)more than(10.81±3.99)% at the control group(P<0.05). Conclusion Pam3CSK4 induce aggregation, activation and the up-regulation of TLR2 of platelet by stimulating TLR2 receptor of it. Thereafter, TLR2 play an important role in the innate immune of platelet.