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(P< 0 .05);PRU of BASE and P2Y12 receptor ,thrombin induced MA value (mm) had no statistical differences between these two groups(P>0 .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 .

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.

Objective To detect the antiplatelet function of clopidogel by Thrombelastography (TEG)and light transmmi-tance aggregometry (LTA)and analyze efficiency of two methods.Methods ①Venous blood samples were taken from 48 outparients and inpatient with acute corotary syndrome (ACS)in PLA General Hospital,including unstable anqina,ST seg-ment elevation myocardial infarction and non ST segment elevation myocardial infarction (32 males,16 females)by random number table from March to July 2011,whose average age was 73 (62~90).②All of them were served 160 mg aspirin and 300 mg clopidogrel after they were in hospital at first time,and then served with 75 mg/d clopidogrel for 9 months.After that,venous blood samples were drew from them who were served clopidogrel 2 hours later.③Platelet aggregation function was assayed with LTA and TEG.All volunteers were signed informed consent and the experiment was approved by the hos-pital ethics committee.Results The platelet aggregation rate detected with LTA induced by adenosine diphosphate (ADPL-TA-INDU)was 40~80% in 41 individuals (85.4%),>80% in 7 individuals (14.6%),coefficient of variation (CV)was 0.19.The inhibition ration of platelet function (INHI)detected with TEG (ADPTEG-INHI)was 10~60% in 40 individu-als (83.3%),<10% in 2 individuals (4.2%),>60 in 6 individuals (12.5%),CV was 0.54.The sum of ADPLTA-INDU and ADPTEG-INHI (ADPLTA-INDU+ADPTEG-INHI)tended to a constant (Mean=98%,SD=12.1).There was no significant indifference between every (ADPLTA-INDU+ADPTEG-INHI)and their Mean (98%)by paired t-test.The ADPLTA-INDU was negative correlated with ADPTEG-INHI (r=-0.75,P<0.01).Conclusion The effect of clopidogrel was mild and the efficiency of TEG and LTA was similar in detecting the antiplatelet function of clopidogrel,but the preci-sion of TEG was lower than LTA.

Objective To investigate the compound donkey-hide gelatin syrup in reducing adverse reactions of qi-blood weakness patients caused by clozapine.Methods 132 patients from Psychiatric Hospital of Yunnan Province between January 2010 and June 2010 were randomly divided into a treatment group and a control group.Both groups were taken clozapine orally.On this basis,the treatment group was taken compound donkey-hide gelatin syrup and the control group was taken placebo syrup.After 8 weeks treatment for both groups,the PANSS,TESS,physical examination and experiment examination were observed to evaluate the clinical efficacy and safety.Results ① the total curative effect:the treatment group was 73.53％,the control group was 65.63％,showing statistical difference (x2=2.543,P＜0.05).② PANSS scores changes before and after the treatment:PANSS score at 2,4,6,8 weeks after the treatment of both groups were [(72.51 ±27.55),(60.54±24.03),(53.12± 15.27),(48.15± 11.88) in treatment group respectively,and (70.71 ±23.90),(58.89± 18.95),(53.06± 14.38),(48.98 ± 9.78) in the control group,respectively],both showing significant difference than the same group before the treatment [(103.99±39.12) in the treatment group,(99.78±34.35) in the control group] (P＜0.05).But there was no statistical significance between two groups (F=2.413,P＞0.05).③ adverse reactions:during the treatment liver function,blood cell analysis,dystonia,Parkinson's obstacle,akathisia,abnormal gastrointestinal reaction,heart rate,heart rate variability and blood pressure in the treatment group was significantly lower than the control group (x2=4.562,P＜0.05).Conclusion Compound donkey-hide gelatin syrup can effectively relieve adverse reactions in qi-blood weak psychosis patients after clozapine treatment and improve their drug tolerance.

Objective To study the way in showing collateral ligaments of knee by MRI and MRI findings of the normal collateral ligaments.Methods MR imaging examinations were performed in 55 healthy volunteers . MR imaging included T1WI and PDWI sequence on the coronal, sagittal and coronal posterior oblique plane, in order to observe the collateral ligaments, and to measure the medial collateral ligament and the lateral collateral ligament dimension, which were compared with the results of the cadaver published in the related anatomic literatures.Results (1)On sagittal MRI, the angle between the line of medial collateral ligament and the tibia long axis was 0.55°±0.25°,the angle between the line of lateral collateral ligament and the long axis of the fibular neck was 11.47°±1.61)°.(2)On coronal MRI, 96% of the medial collateral ligaments was completely showed on only one slice, at the same time, 82% of the lateral collateral ligaments was completely showed on only one slice .On 11°coronal posterior oblique plane,90% of the lateral collateral ligaments was completely showed on only one slice .(3)The normal medial and lateral collateral ligaments were hypointense string on both T1WI and PDWI coronal MR images , with an average length of 11.53 cm for the medial collateral ligament and 5.31 cm for the lateral collateral ligament.Conclusion Using sagittal plane completely shown the tibia and fibular neck as the standard plane and thickness of 3mm in examining the collateral ligaments of knee by MR imaging can display the medial and the lateral collateral ligament clearly .

Objective To determine the prognostic significance of non-thyroidal illness syndrome(NTIS) and FT3 on long-term all-cause and cardiovascular mortality in patients with coronary artery disease(CAD).Methods A total of 1 354 patients underwent coronary angiograhy and participated in the study.After screening,984 patients with CAD were enrolled finally and divided into NTIS group and euthyroid group.The admitted patients were also classified into Tertile 1-3 groups based on FT3 value.The relationship of long-term all-cause and cardiovascular mortality with FT3 was investigated by multivariate Cox regression analysis.Results 129 out of 984 patients had NTIS.As FT3 value decreased,both long-term all-cause mortality (Tertile 1 group 9.6％,Tertile 2 group 11.5％,Tertile 3 group 20.9％,P＜0.01) and cardiovascular mortality (Tertile 1 group 4.5 ％,Tertile 2 group 7.2％,Tertile 3 group 11.0％,P＜0.01) gradually increased.After adjusting for all factors,FT3 (HR =0.614,95％ CI 0.439-0.859)was independently associated with long-term all-cause mortality.FT3 (HR =0.605,95％ CI 0.370-0.986)was also a risk factor for cardiovascular mortality.Conclusion NTIS exists in patients with CAD without myocardial infarction.FT3 reduction is an independent risk factor for long-term all-cause and cardiovascular mortality in patients with CAD.

BACKGROUND:To in vitro isolate neural stem cel s with high purity and uniform biological properties and to establish a complete set of neural stem cel culture system is the basis for neural stem cel research.
OBJECTIVE:To establish an isolation and culture system for neural stem cel s from newborn mouse hippocampus, olfactory bulb and cortex and to analyze the biological properties of cel s.
METHODS:Neural stem cel s were isolated from the hippocampus, olfactory bulb and cortex tissue of newborn Kunming mice by mechanical separation and trypsin digestion. Serum-free culture technology, mechanical pipetting and trypsin digestion were used for subculture of neural stem cel s. 10%fetal bovine serum was used to induce differentiation of neural stem cel s. Neural stem cel s and their differentiated products were identified by
immunofluorescent staining of Nestin, CD133,β-TubulinIII, glial fibril ary acidic protein.
RESULTS AND CONCLUSION:The neural stem cel obtained from newborn mouse hippocampus, olfactory bulb and cortex had the capacity of self-renewal and differentiation which were positive for Nestin and CD133. After induction with fetal bovine serum, neural stem cel could differentiation toβ-tubulinIII or glial fibril ary acidic protein positive cel s that were neurons and astrocytes. This experiment has successful y established the neural stem cel isolation, culture, identification and induction system, providing experimental basis for subsequent studies of neural stem cel s.

Objective The prognosis of patients with acute myocardial infarction (AMI) is related to age,comorbidities,and other factors,in which non-thyroid sick syndrome (NTIS) may also be an important factor.In this study,determination of blood free triiodothyronine (FT3) was used to explore the short-term and long-term mortality relationship of NTIS with acute myocardial infarction.Methods A total of 1 019 cases of newly diagnosed patients with acute myocardial infarction were enrolled.According to FT3 levels,the enlisted subjects were divided into Quartile 1-4 groups; survival group and non-survival group; normal thyroid function and NTIS group.The enrolled subjects were followed-up for 6-90 months,with the median follow-up time of 44.5 months.Using logistic regression and Cox hazards model,the relationships of short-term and long-term mortality in AMI with NTIS or FT3 were compared.Results The incidence of NTIS in patients with AMI was 27.78％.With the progressively decreasing FT3 and FT3/FT4 ratio,the mortality rates were progressively increased (Quartile 1 group 9.4％,Quartile 2 group 13.8％,Quartile 3 group 14.3％,Quartile 4 group 34.0％,P＜0.01).After being adjusted,FT3 was the strongest influencing factor of mortality within 30 days (RR =0.212,95％ CI 0.125-0.359).Multivariate Cox regression analysis showed that FT3 was independently associated with long-term mortality (RR =0.674,95％ CI 0.514-0.885).Kaplan-Meier showed significant difference in mortality between quartile 1-3 groups and the Quartile 4 group.Even FT3 level was within the low normal range,it was related with the mortality in AMI.Conclusions NTIS is common in patients with AMI.After being adjusted,FT3 was the strongest predictor of mortality within 30 days,and low FT3level in AMI patients was an independent risk factor for long-term all-cause mortality.Even FT3 level was within the normal range,it was still related with mortality in myocardial infarction.

Objective To evaluate a new platelet function analyzer PL-11 with three major platelet function methods.Methods A randomized controlled trial was adopted.Totally 20 healthy young students from People's Liberation Army Medical School were enrolled in the study during July and August in 2013.Subjects were treated with aspirin or clopidogrel and the platelet function was measured by using of PL-11,as well as light transmittance aggregometry (LTA),VerifyNow and thromboelastography (TEG).Results When monitor aspirin response,correlations between arachidonic acid (AA) induced PL-11 and other methods were:LTA,0.614; VerifyNow,0.829; TEG,0.697,respectively.Biases between AA induced PL-1 1 and LTA were 1.94％ at baseline and 24.02％ during aspirin treatment.Cut-off value of aspirin response tested with AA induced PL-11 was 33.3％.When monitor clopidogrel response,correlations between adenosine piphosphate (ADP) induced PL-11 and other methods were:LTA,0.767; VerifyNow,0.851; TEG,0.608.Biases between ADP induced PL-11 and LTA were 3.01％ at baseline and 6.56％ during clopidogrel therapy.Cut-off value of clopidogrel response tested with ADP induced PL-1 1 was 66％.Conclusion Results of different platelet function testing methods were not equally effective.PL-11 has a high capability when monitoring short aspirin or clopidogrel response.

Diffuse axonal injury (DAI) is a common primary brain injury.The main mechanism of DAI is the relative displacement of brain tissue junction caused by shearing force during cranial rotation.The pathological changes are the deformation and rupture of white matter nerve fiber bundles.The mechanism of injury and recovery is still unclear,and unified diagnostic criteria are also lacking.Apolipoprotein E (APOE) gene polymorphisms are associated with nervous system development,injury,and repair process.The APOE gene contains six genotypes by the combination among three genes and synthesizes three proteins including apoE2,apoE3 and apoE4.The abnormal spatial structure of apoE4 decreases its ability of transporting lipids,resulting in reduced repair ability after nerve injury.The molecular mechanism of ApoE in the development of axonal injury is complicated.apoE protein can bind to receptor proteins such as low density lipoprotein receptor related protein 1 (LRP1) and heparan sulfate proteoglycan (HSPG) to mediate axonal repair,and it can also damage nerves by influencing calcium influx and producing toxic fragments through hydrolysis.Correcting the abnormal structure of apoE4 is helpful to nerve repair,and apoE analogues also have certain neuroprotective effects.This article reviews the injury characteristics,pathological characteristics,APOE gene polymorphisms,and the role of apoE synthesis in DAI,to provide new insights for elucidation of mechanism of DAI and related clinical application.