Elderly Transient Ischemic Attack Of PAC-1, CD62P And Carotid Atherosclerosis

Background and PurposeBlood in the circulatory system of the human body, blood platelets plays an importantrole in the process of formation of thrombus. Most of them are in the resting State ofplatelet. Platelet activation of platelets in the resting state by a variety of physical, chemicalor biological factors, making platelets are stimulated and activated platelets. Only afteractivation of resting platelets occur adhesion, deformation, aggregation, release of a seriesof process, the formation of plaque, eventually leading to the formation of blood clots.Most major glycoprotein is both PAC-1and CD62P. The physiological role of PAC-1isthe fibrinogen receptor, which is mainly involved in the combination of fibrinogen topromote platelet aggregation, the formation of the PAC-1is a prerequisite for plateletaggregation. CD62P is mediated through blood neutrophils, monocytes and platelets orendothelial cells occurred mutual adhesion, aggregation process, respectively, inthrombosis and inflammation plays an important role in. When after platelet activationoccurs, surface of the platelet glycoprotein expression CD62P markedly enhancedcapability than the original, as markers of Platelet Activation. The experimental detectionof platelet membrane by flow cytometry on the positive expression rate of CD62P, PAC-1,we can understand the level of Platelet Activation.His experiment we tested PAC-1, CD62P level analysis of elderly patients with TIA,and carotid artery color Doppler ultrasound examination, respectively, to observe thechanges in the level of Platelet Activation in elderly patients with TIA and indicators ofchange with carotid atherosclerosis. Study on activation of platelet in TIA of carotidatherosclerosis pathogenesis, reflects the specific molecular markers of Platelet Activationexpression CD62P, PAC-1, and compared with healthy control group, specifically betweenTIA with Platelet Activation and carotid atherosclerosis in the elderly, for prevention ofTIA and TIA, and intervention of progression to find new methods. Materials and methods：Object of study：Cooperation with examination centers in Taian city hospital,2010veteran cadrehealth examination information, Department information, select a normal Checkup of50cases, male32cases,18cases of women with an average age of65.4±6.9years. Used fordetection of platelet-activating protein CD62P, PAC-1, and health groups as TIA. Normalexamination who are excluded past suffers from cardiovascular disease, brain vasculardisease, respiratory disease, digestive system disease, urinary system disease and thehypertension disease, diabetes, tumor, rheumatism immune sexual disease, chronicinfection, has recent surgery, and activities sexual bleeding and the has bleeding tendencies,disease history of examination who, general secondary check, as blood General, liverkidney Gong, blood glucose, lipid, coagulation blood system, obey the General, ECG,chest tablets, check results are is not has exception.Collection of Taian city hospital senile two wards, since the period between October2010and74cases of inpatients with TIA. Transient ischemic attack (transient ischemicattack,TIA) Group (group a)74. Male52cases,22cases of women with an average age of67.4±6.2.Exclusion criteria:1.2weeks before the admission of patients with aspirin, effect ofantiplatelet drugs such as heparin.2. the former suffers from diabetes, acute cerebralvascular disease, cardiac infarction diseases such as atrial fibrillation.3. esophageal andgastric varices, active Ulcerative disease, and so on.4. patients with allergy to aspirin drug.Detection of platelet-activating protein：TIA patients after onset at24h,48h,7d,14d extraction antecubital venous blood,respectively, for the detection of CD62P, PAC-1.Experimental detection method：TIA patients must use a disposable blood taking needle,each pumping venous blood were pierce to the heart of the matter.Subjects were nextmorning after fasting, select the median cubital vein puncture, extraction of venous bloodof about2ml in3.2%sodium citrate anticoagulant blood, blood and anticoagulants in theratio of9:1, gently after mixing, in blood in10minutes using a fluorescein-labeledanti-platelet monoclonal immunofluorescence staining.Fluorescent staining method：1. Of IgG1PE (CD62P PE isotype control) were added in the control tube, PAC-1,CD61PerCP fluorescent antibody20μL of RGDS (PAC-1blocker)10μL.2. Respectively in the test tube PAC-1FITC,CD62P PE,CD61PerCP20μ l offluorescent antibody. Care and joined in a test tube (no activation or activation) blood sample5μ l, after mixing gently, dark and light in the laboratory incubation15-20minutes.3. On the care and test tube is respectively added with precooling of2to8DEG Cfixed in1%paraformaldehyde solution1ml, full mixing, and then placed in2~8c darklight place for30minutes.4.12h inner flow cytometry, CD61PerCP and measuring scattered light scatterdiagram with platelet door, isotype controls set negative groups, determination of plateletmembrane PAC-1, the positive expression of CD62P percentage.5. The same method of detection of50cases of healthy control group of patients withmicro-whole blood platelet activation-specific molecular marker levels of CD62P, PAC-1the percentage of positive expression on the platelet surface.Color Doppler ultrasound detection of caroti：1. Subjects using United States-GE company vivid seven color Doppler ultrasounddetectors, frequency of probe for5-12MHz, carotid artery ultrasonography.2. Designed by an experienced ultrasound technicians are responsible for operationand by color Doppler ultrasound diagnostic imagingCheck method：（1）Subject line before examination by color Doppler ultrasound of the carotid artery,ask subjects to sit or lie to rest about5minutes.（2）Participants check body position to take back, under the shoulder pad pillow, asksubjects to relax, head and neck extension as far as possible, biased on one side, fullyexposed neck.（3）Neck artery color Super check method: will probe gently placed neck roots(clavicle Shang margin), chest lock milk bursting muscle front tracheal next, first forlongitudinal cut sweep check, displayed vascular long axis section, from neck total arterynear heart end along its vascular go line direction to head side mobile, turn displayed necktotal artery (CCA) near end, and middle and distal, to min fork Department respectivelysweep check neck within artery (ICA) and neck outside artery (EcA), vertebral artery (vA)and clavicle Xia artery (SA). As probes into the highest part of the anterior.（4）Color Doppler ultrasound examination in carotid artery: a carotid artery. TIAgroup patients with carotid arterial intima-media thickness (IMT), detection of carotidartery plaque and its properties. Real-time Imaging array probe, frequency is5~12MHz,measurement sites in the carotid artery1cm artery proximal segment of the Department, Department of carotid bifurcation and ball after1cm of the distal internal carotid arterywall, about3points, measuring3times, averaged in. Local vascular cavity defect area rateof blood flow in IMT>1.2mm or10mm2is defined as the carotid artery atheroscleroticplaque; strong echoes for hard spots, low ECHO for soft spots.According to the results of color Doppler ultrasonography, transient cerebral ischemiaattack with CA is divided into sets of TIA and TIA without CA for groups, control groupsare divided into control group with CA and without CA’s controls.Statistically：Use the SAS statistical software for statistical analysis. Each set of data to mean±standard deviation (x±s). Group data using analysis of variance. TIA and PlateletActivation, Pearson correlation analysis of correlation between blood coagulation. P<0.05think there are significant differences, P>0.01think there is a very significant difference.Result：1.TIA in the elderly with normal control group PAC-1, CD62P positive expressionrate of old age PAC-1, TIA group CD62P positive expression rate control groupsignificantly higher than normal, two group comparison difference statistically significant(P<0.01).2.Older TIA, carotid atherosclerosis and healthy control group comparison:(1).TIA group with or without associated with carotid atherosclerosis in the elderlypatients with TIA, PAC-1, CD62P positive expression rate significantly higher than thecorresponding control group; two group comparison difference statistically significant(P<0.01).(2).In elderly patients with TIA or healthy control group, PAC-1, associated withcarotid atherosclerosis CD62P positive rates significantly higher than non-carotidatherosclerosis group, and associated with carotid atherosclerosis in elderly patients withTIA group maximum. Comparison of two groups differences are statistically significant(P<0.01).3.PAC-1and carotid atherosclerotic plaque, CD62P positive expression rate of:associated with carotid atherosclerosis in elderly patients with TIA in the group, groupPAC-l, CD62P atherosclerotic plaque positive rates significantly higher thannon-atherosclerotic plaques set. Comparison of two groups differences are statisticallysignificant (P<0.01).4. Carotid artery atheromatous plaque nature of PAC-l, the positive rate of CD62Pexpression: relationship with carotid atheromatous plaque in elderly TIA group, accompanied by unstable plaque group TIA PAC-1, the positive expression rate of CD62Pwas significantly higher than that in stable plaque group.There was significant differencebetween two groups (P <0.01)Conclusions：In this experimental study, select PAC-1and CD62P as the level of detection ofplatelet activation markers. Experimental results show that TIA CD62P and PAC-1levelsin the group than the normal positive expression of the appropriate control groupsignificantly increased. In the TIA group, PAC-1and CD62P level with the severity ofcarotid intimal lesions gradually increased. This suggests that must exist in PlateletActivation in patients with TIA, and PAC-1and CD62P level with increased carotid arterydamage rises. The reason for this may be after carotid atherosclerosis, vascular endothelialdamage, substances such as collagen and Thrombin generation, prompting plateletmorphology has changed, in the form of calcium and intracellular phosphorylationreactions. Is the second gathering of carotid artery atherosclerotic plaques, forming clumpsthat may cause Vascular Stenosis in the Orwellian newspeak, making blood flow isobstructed, slows, blood flow velocity shear is higher than the normal physiological level,therefore, high shear forces Act allows expression of platelet membrane PAC-1, CD62Plevels play a greater role. This study also found that the level of level of Platelet Activationand carotid atheromatous plaque in the nature of a relationship between them. TIA unstableplaques set of vulnerable plaque with contrast, find PAC-1and CD62P expression clearlyobvious than the latter. The reason for this may be that TIA in patients with unstableplaques on the wall could easily fall off, surface of the platelets adhesion in unstableplaques, platelet activation and the release of PAC-1and CD62P. Prothrombotic State, isthe State of blood coagulation, is because after damage to vascular endothelial cells,platelets and white blood cells in the blood are activated, coagulation factors are activation,fibrinolytic system pathology, blood clots. In short, significantly elevated levels of PlateletActivation in patients with TIA, and with the increase of carotid atherosclerosis in patientswith elevated. Determination of degree of Platelet Activation in patients with TIA helps oncarotid atherosclerosis in judgment, to provide early intervention, reduce TIA recurrentcerebral infarction, prevention has important significance.