ELISA检测D-Dimer、APA、TAT探讨恒古骨伤愈合剂治疗兔TDVT分子机理的实验研究
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摘要
【目的】
1.应用恒古骨伤愈合剂治疗兔创伤性深静脉血栓(TDVT),通过观察血栓形成情况评估其疗效;
2.应用ELISA技术检测D-二聚体(D-Dimer)、抗磷脂抗体(APA)、凝血酶-抗凝血酶复合物(TAT)在恒古骨伤愈合剂治疗兔TDVT三个关键时相点的表达量;
3.通过观察上述三个指标在不同组别的表达变化,探究恒古骨伤愈合剂治疗兔TDVT过程中对凝血系统的影响。
【方法】
一、恒古骨伤愈合剂治疗兔TDVT的疗效观察及血液、血管组织采集
1.实验动物及分组:本实验共64只SPF级新西兰大白兔,按给药不同随机分为正常对照组(A组共8只)、恒古骨伤愈合剂组(B组共28只)、生理盐水对照组(C组共28只);再根据观察时间不同分别将B、C组分为造模后24小时观察组(B_1、C_1组,每组各8只),造模后72小时观察组(B_2、C_2组,每组各8只)及造模后120小时观察组(B_3、C_3组,每组各8只);
2.造模方法:实验采用左侧股静脉钳夹+髋人字石膏固定方法对B、C组兔进行造模。实验兔麻醉后,取腹股沟内侧沿股动脉走行方向作一长4-5cm纵行切口,暴露约2.5cm的左侧股静脉,用12.5mm全齿蚊式血管钳分五段各钳夹1次,力量为血管钳紧2扣,每次持续3秒;用1号丝线全层间断缝合皮肤切口,不放置引流,均行石膏固定;
3.给药时间及剂量:B组兔造模后立刻灌喂恒古骨伤愈合剂,以后每隔24h喂药一次(剂量为1.17ml/kg);C组兔在对应时间灌喂生理盐水(剂量为1.17ml/kg);
4.观察、取材时间及方法:造模前对A组各兔抽取耳缘静脉血1.5ml,装入标准枸橼酸钠抗凝管,混匀、离心、取血浆-20℃保存;造模后24小时,B组随机取8只作为B_1组,C组随机取8只作为C_1组,剪开切口缝线,肉眼观察并比较两组血栓发生率,并取长约1.0cm的股静脉进行HE染色、光镜观察确定血栓形成状态,同时,由耳缘静脉抽取每只兔各1.5ml静脉血,装入标准枸橼酸钠抗凝管,混匀,3000rpm/min离心20min后取血浆-20℃保存;造模后72小时和120小时分别随机取8只作为B_2、C_2、B_3、C_3组,以相同方式取材、采血;
5.血栓发生率统计学处理:对各组资料数据进行x~2检验,以P＜0.05为有统计学意义,P＜0.01为有显著的统计学意义。
二、ELISA检测D-Dimer、APA、TAT探讨恒古骨伤愈合剂治疗TDVT对凝血系统的影响
1.ELISA检测:采用ELISA方法分别检测A、B_1、B_2、B_3、C_1、C_2、C_3各组大白兔血液中D-Dimer、APA、TAT的表达量;
2.统计学处理:使用SPSS11.5统计软件包对所得表达量进行统计学处理。各组计量资料数据用均数士标准差((?)±s)表示,两组数据间比较用单因素方差分析(One-way ANOVA)和S-N-K检验,以P＜0.05为差别有统计学意义,P＜0.01为差别有显著的统计学意义。
【结果】
一、恒古骨伤愈合剂治疗兔TDVT的疗效观察及血液样本采集
1.实验动物一般情况:B、C两组兔共死亡5只,动物死亡率8.93%
2.血栓发生情况:该模型中,股静脉血栓形成高峰期在造模后24h,血栓发生率分别为:B组66.7%(16/24),C组79.2%(22/24),差别有统计学意义(P＜0.05);之后血栓开始消退,在恒古骨伤愈合剂治疗72h,血栓可基本消退,此时血栓发生率为:B组25.0%(4/16),C组75.0%(12/16),差别有统计学意义(P＜0.05);造模后120h,B_3组血栓已完全消退,C_3组血栓发生率为50%(4/8),两者比较差别有显著的统计学意义(p＜0.01)。
3.各组所获取的股静脉组织,肉眼、光镜观察结果与相应血栓状态一致。
二、ELISA检测D-Dimer、APA、TAT探讨恒古骨伤愈合剂治疗TDVT对凝血系统的影响
1.C组(9.11±1.11ng/ml)与A组(1.70±0.30ng/ml)相比,D-dimer、APA、TAT的表达量存在统计学差异
2.C组(9.11±1.11ng/ml)与B组(2.27±0.62 ng/ml)相比,D-dimer、APA、TAT的表达量存在统计学差异
【结论】
1.恒古骨伤愈合剂能有效降低兔TDVT发生率,促进血栓消退;
2.恒古骨伤愈合剂可能通过影响凝血系统,降低血液高凝状态治疗兔TDVT。
Objective.
1.To explore therapeutic effect of Osteoking in treatment of rabbits TDVT by observe thrombosis situation.
2.After treatment with Osteoking in the rabbits TDVT.ELISA applied to detected expression levels of D-Dimmer、APA、TAT at different phases of rabbits TDVT.
3.By observing the changes of D-Dimmer,APA,TAT during the different group of traumatic deep vein thrombosis(TDVT),to explore the role of fibrinolytic and coagulation systems during the treatment with Osteoking.
Methods.
1.Therapeutic effect observation with Osteoking in rabbits TDVT and collection of blood and vascular tissue samples
Animal and grouping:According to different drug treated with Osteoking,64 SPF rabbits were divided randomly into control group(group A,total 8),Osteoking group(group B,total 28),physiological saline group(group B,total 28).According to different observation phases B,C groups were divided into other 6 groups: post-modeling 24h group(B_1,C_1 group,respectively 8),post-modeling 72h group(B_2,C_2 group,respectively 8) and post-modeling 120h group(B_3,C_3 group, respectively 8).
Modeling:Directly clamping left femoral vein combined with hip spica fixation was performed to establish acute TDVT rabbits models with B,C groups.Rabbits were anesthetized,supine position fixation,sterilization,exposing left femoral veins.2.5cm isolated femoral vein was clamped at five points separately with 12.5mm mosquito-hemostatic forceps,once each point;the clamping strength was fastening two barb of hemostatic forceps,lasting 3 seconds each time,then the incision was sutured,no drainage was set,left posterior limbs were fixed with hip spica casts.
Administration and dosage:Once every 24 hours,administrate in group B with Osteoking about 1.17ml/kg,while C group with normal saline with the same dosage.
Observation,methods and time for obtaining blood and vein tissue:To collect A group rabbits blood about 1.5ml from ear-edge vein at 24 hours after modeling 8 rabbits were selected randomly from group B to establish B_1 group,while the rabbits were selected randomly from group B to establish B_1 group,then resect the suture of the wound gross observe ratio of the left femoral vein thrombosis between the two groups.Resected 1.0cm vascular tissue put into stationary liquid for the further HE staining histological analysis and light microscope.8 rabbits were selected randomly at post-modeling 72h and 120h to establish group B_2,C_2,B_3,C_3 respectively,then obtain blood and vascular specimen for the same method of group B_1.
Statistical analysis about TDVT incidence rate:Information on the data in each group using x~2 test,P＜0.05 for statistically significant,P＜0.01 for significant statistical significance.
2.ELISA detection of D-dimer,APA,TAT to explore Osteoking treatmentTDVT impact on the coagulation system
ELISA were used to detect the blood D-dimer,APA,TAT expression of rabbits in A、B_1、B_2、B_3、C_1、C_2、C_3 group.
Using SPSS 11.5 statistical software package analyze the expression of the statistical volume.Measurement data in each group are demonstrate by the standard deviation(±s),compare with two sets of data using one-way ANOVA and SNK test, P＜0.05 for the differences were statistically significant,P＜0.01 for the significant difference in statistical significance.
Results
1.The condition of rabbits:5 rabbits died in group B,C,and mortality is 8.93%.
Thrombosis:In this model,post-modeling 24h is the thrombotic crest-time of femoralvein,the average incidence of TDVT between the two groups is 80.8%,then thrombosus begin spontaneous to resolution.After Osteoking treatment thrombosus almost resolution.After modeling 24h left femoral vein DVT incidence rate:Group B are 66.7%,Group C 79.2%,This had statistical significance(p＜0.05);After modeling 72h Group B are25.0%,Group C are 75.0%also had statistical significance(p＜0.05). After modeling 72h Group B_3 almost resolution.Group B_3 are 50%the two groups comparison with each other has significant statistical significance.(p＜0.01)
Gross observation and light microscope observation of vascular specimens of each group consistent with the corresponding state thrombosis.
2.ELISA detection of D-dimer,APA,TAT to explore Osteoking treatmentTDVT impact on the coagulation system.
D-dimer,APA,TAT expression of group C(9.11±1.11ng/ml) comparison with group A(1.70±0.30ng/ml) exsist significant statistical different.
D-dimer,APA,TAT expression of group C(9.11±1.11ng/ml) comparison with group B(2.27±0.62 ng/ml)exsist significant statistical different.
Conlusion.
1.Treatment with Osteoking in the rabbits TDVT could decrease the incidence of thrombosis and promote thrombolysis.
2.Osteoking may affect the coagulation system,reducing blood hypercoagulation in treated rabbits TDVT.
1.应用恒古骨伤愈合剂治疗兔创伤性深静脉血栓(TDVT),通过观察血栓形成情况评估其疗效;
2.应用ELISA技术检测D-二聚体(D-Dimer)、抗磷脂抗体(APA)、凝血酶-抗凝血酶复合物(TAT)在恒古骨伤愈合剂治疗兔TDVT三个关键时相点的表达量;
3.通过观察上述三个指标在不同组别的表达变化,探究恒古骨伤愈合剂治疗兔TDVT过程中对凝血系统的影响。
【方法】
一、恒古骨伤愈合剂治疗兔TDVT的疗效观察及血液、血管组织采集
1.实验动物及分组:本实验共64只SPF级新西兰大白兔,按给药不同随机分为正常对照组(A组共8只)、恒古骨伤愈合剂组(B组共28只)、生理盐水对照组(C组共28只);再根据观察时间不同分别将B、C组分为造模后24小时观察组(B_1、C_1组,每组各8只),造模后72小时观察组(B_2、C_2组,每组各8只)及造模后120小时观察组(B_3、C_3组,每组各8只);
2.造模方法:实验采用左侧股静脉钳夹+髋人字石膏固定方法对B、C组兔进行造模。实验兔麻醉后,取腹股沟内侧沿股动脉走行方向作一长4-5cm纵行切口,暴露约2.5cm的左侧股静脉,用12.5mm全齿蚊式血管钳分五段各钳夹1次,力量为血管钳紧2扣,每次持续3秒;用1号丝线全层间断缝合皮肤切口,不放置引流,均行石膏固定;
3.给药时间及剂量:B组兔造模后立刻灌喂恒古骨伤愈合剂,以后每隔24h喂药一次(剂量为1.17ml/kg);C组兔在对应时间灌喂生理盐水(剂量为1.17ml/kg);
4.观察、取材时间及方法:造模前对A组各兔抽取耳缘静脉血1.5ml,装入标准枸橼酸钠抗凝管,混匀、离心、取血浆-20℃保存;造模后24小时,B组随机取8只作为B_1组,C组随机取8只作为C_1组,剪开切口缝线,肉眼观察并比较两组血栓发生率,并取长约1.0cm的股静脉进行HE染色、光镜观察确定血栓形成状态,同时,由耳缘静脉抽取每只兔各1.5ml静脉血,装入标准枸橼酸钠抗凝管,混匀,3000rpm/min离心20min后取血浆-20℃保存;造模后72小时和120小时分别随机取8只作为B_2、C_2、B_3、C_3组,以相同方式取材、采血;
5.血栓发生率统计学处理:对各组资料数据进行x~2检验,以P＜0.05为有统计学意义,P＜0.01为有显著的统计学意义。
二、ELISA检测D-Dimer、APA、TAT探讨恒古骨伤愈合剂治疗TDVT对凝血系统的影响
1.ELISA检测:采用ELISA方法分别检测A、B_1、B_2、B_3、C_1、C_2、C_3各组大白兔血液中D-Dimer、APA、TAT的表达量;
2.统计学处理:使用SPSS11.5统计软件包对所得表达量进行统计学处理。各组计量资料数据用均数士标准差((?)±s)表示,两组数据间比较用单因素方差分析(One-way ANOVA)和S-N-K检验,以P＜0.05为差别有统计学意义,P＜0.01为差别有显著的统计学意义。
【结果】
一、恒古骨伤愈合剂治疗兔TDVT的疗效观察及血液样本采集
1.实验动物一般情况:B、C两组兔共死亡5只,动物死亡率8.93%
2.血栓发生情况:该模型中,股静脉血栓形成高峰期在造模后24h,血栓发生率分别为:B组66.7%(16/24),C组79.2%(22/24),差别有统计学意义(P＜0.05);之后血栓开始消退,在恒古骨伤愈合剂治疗72h,血栓可基本消退,此时血栓发生率为:B组25.0%(4/16),C组75.0%(12/16),差别有统计学意义(P＜0.05);造模后120h,B_3组血栓已完全消退,C_3组血栓发生率为50%(4/8),两者比较差别有显著的统计学意义(p＜0.01)。
3.各组所获取的股静脉组织,肉眼、光镜观察结果与相应血栓状态一致。
二、ELISA检测D-Dimer、APA、TAT探讨恒古骨伤愈合剂治疗TDVT对凝血系统的影响
1.C组(9.11±1.11ng/ml)与A组(1.70±0.30ng/ml)相比,D-dimer、APA、TAT的表达量存在统计学差异
2.C组(9.11±1.11ng/ml)与B组(2.27±0.62 ng/ml)相比,D-dimer、APA、TAT的表达量存在统计学差异
【结论】
1.恒古骨伤愈合剂能有效降低兔TDVT发生率,促进血栓消退;
2.恒古骨伤愈合剂可能通过影响凝血系统,降低血液高凝状态治疗兔TDVT。
Objective.
1.To explore therapeutic effect of Osteoking in treatment of rabbits TDVT by observe thrombosis situation.
2.After treatment with Osteoking in the rabbits TDVT.ELISA applied to detected expression levels of D-Dimmer、APA、TAT at different phases of rabbits TDVT.
3.By observing the changes of D-Dimmer,APA,TAT during the different group of traumatic deep vein thrombosis(TDVT),to explore the role of fibrinolytic and coagulation systems during the treatment with Osteoking.
Methods.
1.Therapeutic effect observation with Osteoking in rabbits TDVT and collection of blood and vascular tissue samples
Animal and grouping:According to different drug treated with Osteoking,64 SPF rabbits were divided randomly into control group(group A,total 8),Osteoking group(group B,total 28),physiological saline group(group B,total 28).According to different observation phases B,C groups were divided into other 6 groups: post-modeling 24h group(B_1,C_1 group,respectively 8),post-modeling 72h group(B_2,C_2 group,respectively 8) and post-modeling 120h group(B_3,C_3 group, respectively 8).
Modeling:Directly clamping left femoral vein combined with hip spica fixation was performed to establish acute TDVT rabbits models with B,C groups.Rabbits were anesthetized,supine position fixation,sterilization,exposing left femoral veins.2.5cm isolated femoral vein was clamped at five points separately with 12.5mm mosquito-hemostatic forceps,once each point;the clamping strength was fastening two barb of hemostatic forceps,lasting 3 seconds each time,then the incision was sutured,no drainage was set,left posterior limbs were fixed with hip spica casts.
Administration and dosage:Once every 24 hours,administrate in group B with Osteoking about 1.17ml/kg,while C group with normal saline with the same dosage.
Observation,methods and time for obtaining blood and vein tissue:To collect A group rabbits blood about 1.5ml from ear-edge vein at 24 hours after modeling 8 rabbits were selected randomly from group B to establish B_1 group,while the rabbits were selected randomly from group B to establish B_1 group,then resect the suture of the wound gross observe ratio of the left femoral vein thrombosis between the two groups.Resected 1.0cm vascular tissue put into stationary liquid for the further HE staining histological analysis and light microscope.8 rabbits were selected randomly at post-modeling 72h and 120h to establish group B_2,C_2,B_3,C_3 respectively,then obtain blood and vascular specimen for the same method of group B_1.
Statistical analysis about TDVT incidence rate:Information on the data in each group using x~2 test,P＜0.05 for statistically significant,P＜0.01 for significant statistical significance.
2.ELISA detection of D-dimer,APA,TAT to explore Osteoking treatmentTDVT impact on the coagulation system
ELISA were used to detect the blood D-dimer,APA,TAT expression of rabbits in A、B_1、B_2、B_3、C_1、C_2、C_3 group.
Using SPSS 11.5 statistical software package analyze the expression of the statistical volume.Measurement data in each group are demonstrate by the standard deviation(±s),compare with two sets of data using one-way ANOVA and SNK test, P＜0.05 for the differences were statistically significant,P＜0.01 for the significant difference in statistical significance.
Results
1.The condition of rabbits:5 rabbits died in group B,C,and mortality is 8.93%.
Thrombosis:In this model,post-modeling 24h is the thrombotic crest-time of femoralvein,the average incidence of TDVT between the two groups is 80.8%,then thrombosus begin spontaneous to resolution.After Osteoking treatment thrombosus almost resolution.After modeling 24h left femoral vein DVT incidence rate:Group B are 66.7%,Group C 79.2%,This had statistical significance(p＜0.05);After modeling 72h Group B are25.0%,Group C are 75.0%also had statistical significance(p＜0.05). After modeling 72h Group B_3 almost resolution.Group B_3 are 50%the two groups comparison with each other has significant statistical significance.(p＜0.01)
Gross observation and light microscope observation of vascular specimens of each group consistent with the corresponding state thrombosis.
2.ELISA detection of D-dimer,APA,TAT to explore Osteoking treatmentTDVT impact on the coagulation system.
D-dimer,APA,TAT expression of group C(9.11±1.11ng/ml) comparison with group A(1.70±0.30ng/ml) exsist significant statistical different.
D-dimer,APA,TAT expression of group C(9.11±1.11ng/ml) comparison with group B(2.27±0.62 ng/ml)exsist significant statistical different.
Conlusion.
1.Treatment with Osteoking in the rabbits TDVT could decrease the incidence of thrombosis and promote thrombolysis.
2.Osteoking may affect the coagulation system,reducing blood hypercoagulation in treated rabbits TDVT.
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[18].徐淑云.药理实验方法(第三版):202-204
[19].Thomas DP,Merton RE,vessel wall injury andWood RE,et al.The venous thrombosis relationship between:An experimental study.Br J Haematol,1985,59(1):449-457.
[20].荆志成,韩志岩.静脉血栓形成研究动向.中华结核和呼吸杂志,2000,23(9):522-523.
[21].Meissner MH,Chandler WL,Elliott JS.Venous thromboembolism in trauma:a local manifestation of systemic hypercoagadability.J Trauma,2003,54(2):224-231.
[22].George C,Velmahos MD.Prevention of venous thromboembolism after injury-An evidence-based report-part Ⅱ:analysts of risk factors and evaluation of the tole of vena caval filters.) Trauma.2000,49:140-144
[23].尚德俊,侯玉芬,陈柏楠.周围静脉疾病学[M].北京:人民军医出版,2001:25-26
[24].汪忠镐.重视血管外科的基本原则.中华实用外科杂志 2000;20(6):323
[25].毕书祝,朱红岩.下肢深静脉血栓形成的中西医结合治疗.临床和实验医学杂志2007,6(2):121
[26].金星,秦红松,尚德俊,中西医结合治疗下肢深静脉血栓形成206例临床观察,中国中西医结合杂志,1997,17(5):267
[27].崔书克,刘成藏.中西医结合治疗下肢深静脉血栓形成120例[J].河南中医,2002,22(2):43-44.
[28].史以菊,吴俊芳,夏作理等.三七总皂甙对大鼠局灶性脑缺血的保护作用.中国临床药理学与治疗学杂志,1999:4(4):272
[29].简道林,余金甫,黄海波等.三七总皂甙对缺血一再灌注兔脑保护作用机制的研究.中国危重病急救医学.1999,11(3):145
[30].詹合琴,李平法,杨锦南等.三七皂苷Rgl对t-PA和PA.I-1水平的影响.中国药理学通报,2006;22(7):869-72
[31].许军,王阶,温林军三七总皂苷干预血栓形成研究概况.云南中医中药杂志,2003;24 (5):46-47
[32].白玉莲,王美玲,刘景德.红花对冠心病患者血液流变学和临床的影响.中医药研究,1991,5(5):33.
[33].黄正良,高其明,崔祝梅,红花黄素的药理研究.中草药.1984,15(8):12
[34].ShitritD,Heyd J,Raveh D,et al.Diagiostic value of the D-dimer testin deep vein thrombosis:imp roved results by a new assay method and by using discriminate levels[J].Thromb Res,2001,102(2):125-131
[35].罗慰慈.现代呼吸病学[M].北京:人民军医出版社,1997.
[36].肖慧芳.抗凝血酶治疗弥漫性血管内凝血的研究进展.2008 14(1):36-37
[37].EISELE B,LAMYM,TH IJS LG,et al.Antithrombin Ⅲ in patients with severe sepsis.A randomized,placebo-controlled,double-blind multicenter trial plus a meta-analysis on all randomized,placebo-controlled,double2blind trialswith antithrombin Ⅲ in severe sep sis[J].Intensive CareMed,1998,24(7):663-672
[38].INTHORN D,HOFFMANN JN,HATL WH,et al.Effect of antithrombin Ⅲ supp lementation on inflammatory response in patients with severe sep sis[J].Shock,1998,10(2):90-96.
[39].高飞,章金勇.严重创伤患者凝血纤溶特点分析.重庆医学,2008:37(2):258-259.
[40].NIESSEN RW,LAMPING RJ,JANSEN PM,et al.Antithrombin acts as a negative acute phase protein as established with studies on HepG2 cells and in baboons[J].Thromb Haemost,1997,78(3):1088-1092.
[41].ESMON CT.Are nature anticoaguLants candidates formodulating the inflammatory response to endotoxin[J].Blood,2000,95:1113-1116
[42].芦璐,侯明,高阳等,深静脉血栓形成患者抗活化蛋白C与抗磷脂抗体相关性分析.临床血液学杂志.2007,5(20)3:149-151
[43].Schorer AE,Wickham NW,Watson KV,et al,Lupus anticoagulant induces a selective defect in thrombin -mediated endothelial prostacyclin release and platelet aggregation[J].Br J Haematol,1989,71(3):399-407.
[44].邓宇斌.抗磷脂抗体的共辅因子β 2GPI的研究进展[J].上海免疫学杂志,1997,17(1):61-62.
[45].Ilahi OA,Reddy J,Ahmad I.Deep venous thrombosis after knee arthroscopy:a meta-analysis.Arthroscopy.2005 Jun;21(6):727-730.
[1].Greenfield LJ,He AR,Henke PK,Hingorani A,Hull RD,Kessler CM,McBane RD,McLafferty R.Acute venous disease:venous thrombosis and venous trauma.J Vasc Surg.2007 Dec;46 Suppl S:25S-53S.
[2].Rosendaal FR,VAN Hylckama Vlieg A,Doggen CJ.Venous thrombosis in the elderly.J Thromb Haemost.2007 Jul;5 Suppl 1:310-7.
[3].Goldhaber SZ,Visani L,De Rosa M.Acute pulmonary embolism:clinical outcomes in the International Cooperative Pulmonary Emholism Registry(ICOPER).Lancet,1999,353:1386-1389
[4].HeitJA,Cohen AT,Anderson FA,VTE Impact Assessment Group.Estimated annual number of incident and recurrent,nonfatal and fatal venous thromboembolism(VTE) events in the U.S.Blood 2005;1OC:27.
[5].Brotman DJ,Deitcher SR,Lip GY,et al.Virchow's triad revisited.South Med J.2004;97(2):213-4
[6].潘家绮,张之南.血栓形成的分子标志物及其临床意义.中华内科杂志,1990,29(7).395-397.
[7].朱广谨,周朝凤.纤溶系统激活的实验研究——儿茶酚胺对纤溶活性的影响.中国病理生理杂志,1991,7(3):267-271
[8].Rosendaal FR.Venous thrombosis:a multicausal disease.Lancet,1999,353:1167-1173
[9].Kearon C.Epidemiology of venous thromboembolism.Semin Vasc Med,2001,1:7-26.
[10].荆志成,韩志岩.静脉血栓形成研究动向.中华结核和呼吸杂志,2000,23(9):522-523.
[11].Meissner MH,Chandler WL,Elliott JS.Venous thromboembolism in trauma:a local manifestation of systemic hypercoagadability.J Trauma,2003,54(2):224-231.
[12].George C,Velmahos MD.Prevention of venous thromboembolism after injury-An evidence-based report-part Ⅱ:analysts of risk factors and evaluation of the tole of vena caval filters.) Trauma.2000,49:140-144.
[13].Michelle M,Gear H,Pharm D,et al.The risk assessment profile score identifies trauma patients at risk for deep vein thrombosis.Surg,2000.128:631-640.
[14].尚德俊,侯玉芬,陈柏楠.周围静脉疾病学[M].北京:人民军医出版,2001:25-26
[15].张强.抗凝药物在血管外科中的应用.中华实用外科杂志.2000;20(6):327
[16].龙景培,胡铭荣,王雄等.下肢深静脉血栓形成23例.医学理论与实践.2000;13(1):35
[17].Greenfield LJ.Current status of surgical therapy for deep vein thrombosis.Am J Surg 1985;10(suppl):64
[18].汪忠镐.重视血管外科的基本原则.中华实用外科杂志 2000;20(6):323
[19].金星,秦红松,尚德俊,中西医结合治疗下肢深静脉血栓形成206例临床观察,中国中西医结合杂志,1997,17(5):267
[20].高德,唐祖宣.《伤寒论》诊疗程序临床验证105例小结,1995,11(3):31-32
[21].王景春,王志平.辨治髂股静脉血栓形成210例.辽宁中医杂志,1996,23(6):265-265
[22].李玉幸,王凤莲.活血化瘀法治疗下肢深静脉血栓形成134例临床及实验观察.1997;19(1):13-14
[23].尚德俊,张秀英.中西医结合治疗下肢深静脉血栓形成研究进展.山东中医药大学学报.2000,24(4):309-313
[24].崔书克,刘成藏.中西医结合治疗下肢深静脉血栓形成120例[J].河南中医,2002,22(2):43-44.
[25].史以菊,吴俊芳,夏作理等.三七总皂甙对大鼠局灶性脑缺血的保护作用.中国临床药理学与治疗学杂志,1999,4(4):272
[26].简道林,余金甫,黄海波等.三七总皂甙对缺血再灌注兔脑保护作用机制的研究.中国危重病急救医学.1999,11(3):145
[27].詹合琴,李平法,杨锦南等.三七皂苷Rg1对t-PA和PA I-1水平的影响.中国药理学通报,2006;22(7):869-72
[28].许军,王阶,温林军三七总皂苷干预血栓形成研究概况.云南中医中药杂志,2003;24(5):46-47
[29].白玉莲,王美玲,刘景德.红花对冠心病患者血液流变学和临床的影响.中医药研究,1991,5(5):33.
[30].黄正良,高其明,崔祝梅,红花黄素的药理研究.中草药.1984,15(8):12Meissner MH,Wakefield TW,Ascher E,Caprini JA,Comerota AJ,Eklof B,Gillespie DL.
[2].付妍,王大为.血栓栓塞性疾病的急诊治疗临床医生和预防.中国.2006,34(8):51-53.
[3].邱贵兴,戴魁戎,杨庆铭,等.预防骨科大手术后深静脉血栓形成的专家建议—深静脉血栓形成预防座谈会纪要.中华骨科杂志.2005;25(10):636-640.
[4].Piovella F,Wang CJ,Lu H,et al.Deep-vein thrombosis rates after major orthopedic surgery in Asia.An epidemiological study based on postoperative screening with centrally adjudicated bilateral venography.Thromb Haemost.2005,11;3(12):2664-2670.
[5].John A.Heit.The Epidemiology of thromboembolism in the community.[J]Arteriosclerosis,Thromb,Vasc,Biol.2008;28:370-372.
[6].Fedullo PF,Auger WR,Channick RN,et al.Chronic thromboembolic pulmonary hypertension.Clin Chest Med.2001;22:561-581
[7].Cohen AT,Agnelli G,Anderson FA,et al.Venous thromboembolism(VTE) in Europe.The number of VTE events and associated morbidity and mortality.Thromb Haemost.2007,10;98(4):756-764.
[8].庄金满.下肢深静脉血栓形成的治疗.中国微创外科杂志.2008年4月8(4):379-380
[9].Thiagarajan P,Wu KK.Mechanisms of antithrombotic drugs.Adv Pharmacol.2000,46:297-324.
[10].Comerota AJ,Paolini D.Treatment of acute iliofemoral deep venous thrombosis:a strategy of thrombus removal.Eur J Vasc Endovasc Surg.2007,33(3):351-60;discussion 361-362.
[11].Bulger CM,Jacobs C,Patel NH.Epidemiology of acute deep vein thrombosis:Tech Vasc Interv Radiol.2004,7(2):50-54.
[12].ShitritD,Heyd J,Raveh D,et al.Diagiostic value of the D-dimer testin deep vein thrombosis:imp roved results by a new assay method and by using discriminate levels[J].Thromb Res,2001,102(2):125-131
[13].罗慰慈.现代呼吸病学[M].北京:人民军医出版社,1997
[14].肖慧芳.抗凝血酶治疗弥漫性血管内凝血的研究进展.2008 14(1):36-37
[15].芦璐,侯明,高阳等,深静脉血栓形成患者抗活化蛋白C与抗磷脂抗体相关性分析.临床 血液学杂志.2007.5(20)3:149-151
[16].赵学凌.创伤性肢体深静脉血栓形成的新型动物模型建立及相关研究.博士学位论文,2004,7
[17].王兵.急性创伤性肢体深静脉血栓大鼠动物模型的建立及局部静脉血管基因表达谱变化.博士学位论文,2005,5
[18].徐淑云.药理实验方法(第三版):202-204
[19].Thomas DP,Merton RE,vessel wall injury andWood RE,et al.The venous thrombosis relationship between:An experimental study.Br J Haematol,1985,59(1):449-457.
[20].荆志成,韩志岩.静脉血栓形成研究动向.中华结核和呼吸杂志,2000,23(9):522-523.
[21].Meissner MH,Chandler WL,Elliott JS.Venous thromboembolism in trauma:a local manifestation of systemic hypercoagadability.J Trauma,2003,54(2):224-231.
[22].George C,Velmahos MD.Prevention of venous thromboembolism after injury-An evidence-based report-part Ⅱ:analysts of risk factors and evaluation of the tole of vena caval filters.) Trauma.2000,49:140-144
[23].尚德俊,侯玉芬,陈柏楠.周围静脉疾病学[M].北京:人民军医出版,2001:25-26
[24].汪忠镐.重视血管外科的基本原则.中华实用外科杂志 2000;20(6):323
[25].毕书祝,朱红岩.下肢深静脉血栓形成的中西医结合治疗.临床和实验医学杂志2007,6(2):121
[26].金星,秦红松,尚德俊,中西医结合治疗下肢深静脉血栓形成206例临床观察,中国中西医结合杂志,1997,17(5):267
[27].崔书克,刘成藏.中西医结合治疗下肢深静脉血栓形成120例[J].河南中医,2002,22(2):43-44.
[28].史以菊,吴俊芳,夏作理等.三七总皂甙对大鼠局灶性脑缺血的保护作用.中国临床药理学与治疗学杂志,1999:4(4):272
[29].简道林,余金甫,黄海波等.三七总皂甙对缺血一再灌注兔脑保护作用机制的研究.中国危重病急救医学.1999,11(3):145
[30].詹合琴,李平法,杨锦南等.三七皂苷Rgl对t-PA和PA.I-1水平的影响.中国药理学通报,2006;22(7):869-72
[31].许军,王阶,温林军三七总皂苷干预血栓形成研究概况.云南中医中药杂志,2003;24 (5):46-47
[32].白玉莲,王美玲,刘景德.红花对冠心病患者血液流变学和临床的影响.中医药研究,1991,5(5):33.
[33].黄正良,高其明,崔祝梅,红花黄素的药理研究.中草药.1984,15(8):12
[34].ShitritD,Heyd J,Raveh D,et al.Diagiostic value of the D-dimer testin deep vein thrombosis:imp roved results by a new assay method and by using discriminate levels[J].Thromb Res,2001,102(2):125-131
[35].罗慰慈.现代呼吸病学[M].北京:人民军医出版社,1997.
[36].肖慧芳.抗凝血酶治疗弥漫性血管内凝血的研究进展.2008 14(1):36-37
[37].EISELE B,LAMYM,TH IJS LG,et al.Antithrombin Ⅲ in patients with severe sepsis.A randomized,placebo-controlled,double-blind multicenter trial plus a meta-analysis on all randomized,placebo-controlled,double2blind trialswith antithrombin Ⅲ in severe sep sis[J].Intensive CareMed,1998,24(7):663-672
[38].INTHORN D,HOFFMANN JN,HATL WH,et al.Effect of antithrombin Ⅲ supp lementation on inflammatory response in patients with severe sep sis[J].Shock,1998,10(2):90-96.
[39].高飞,章金勇.严重创伤患者凝血纤溶特点分析.重庆医学,2008:37(2):258-259.
[40].NIESSEN RW,LAMPING RJ,JANSEN PM,et al.Antithrombin acts as a negative acute phase protein as established with studies on HepG2 cells and in baboons[J].Thromb Haemost,1997,78(3):1088-1092.
[41].ESMON CT.Are nature anticoaguLants candidates formodulating the inflammatory response to endotoxin[J].Blood,2000,95:1113-1116
[42].芦璐,侯明,高阳等,深静脉血栓形成患者抗活化蛋白C与抗磷脂抗体相关性分析.临床血液学杂志.2007,5(20)3:149-151
[43].Schorer AE,Wickham NW,Watson KV,et al,Lupus anticoagulant induces a selective defect in thrombin -mediated endothelial prostacyclin release and platelet aggregation[J].Br J Haematol,1989,71(3):399-407.
[44].邓宇斌.抗磷脂抗体的共辅因子β 2GPI的研究进展[J].上海免疫学杂志,1997,17(1):61-62.
[45].Ilahi OA,Reddy J,Ahmad I.Deep venous thrombosis after knee arthroscopy:a meta-analysis.Arthroscopy.2005 Jun;21(6):727-730.
[1].Greenfield LJ,He AR,Henke PK,Hingorani A,Hull RD,Kessler CM,McBane RD,McLafferty R.Acute venous disease:venous thrombosis and venous trauma.J Vasc Surg.2007 Dec;46 Suppl S:25S-53S.
[2].Rosendaal FR,VAN Hylckama Vlieg A,Doggen CJ.Venous thrombosis in the elderly.J Thromb Haemost.2007 Jul;5 Suppl 1:310-7.
[3].Goldhaber SZ,Visani L,De Rosa M.Acute pulmonary embolism:clinical outcomes in the International Cooperative Pulmonary Emholism Registry(ICOPER).Lancet,1999,353:1386-1389
[4].HeitJA,Cohen AT,Anderson FA,VTE Impact Assessment Group.Estimated annual number of incident and recurrent,nonfatal and fatal venous thromboembolism(VTE) events in the U.S.Blood 2005;1OC:27.
[5].Brotman DJ,Deitcher SR,Lip GY,et al.Virchow's triad revisited.South Med J.2004;97(2):213-4
[6].潘家绮,张之南.血栓形成的分子标志物及其临床意义.中华内科杂志,1990,29(7).395-397.
[7].朱广谨,周朝凤.纤溶系统激活的实验研究——儿茶酚胺对纤溶活性的影响.中国病理生理杂志,1991,7(3):267-271
[8].Rosendaal FR.Venous thrombosis:a multicausal disease.Lancet,1999,353:1167-1173
[9].Kearon C.Epidemiology of venous thromboembolism.Semin Vasc Med,2001,1:7-26.
[10].荆志成,韩志岩.静脉血栓形成研究动向.中华结核和呼吸杂志,2000,23(9):522-523.
[11].Meissner MH,Chandler WL,Elliott JS.Venous thromboembolism in trauma:a local manifestation of systemic hypercoagadability.J Trauma,2003,54(2):224-231.
[12].George C,Velmahos MD.Prevention of venous thromboembolism after injury-An evidence-based report-part Ⅱ:analysts of risk factors and evaluation of the tole of vena caval filters.) Trauma.2000,49:140-144.
[13].Michelle M,Gear H,Pharm D,et al.The risk assessment profile score identifies trauma patients at risk for deep vein thrombosis.Surg,2000.128:631-640.
[14].尚德俊,侯玉芬,陈柏楠.周围静脉疾病学[M].北京:人民军医出版,2001:25-26
[15].张强.抗凝药物在血管外科中的应用.中华实用外科杂志.2000;20(6):327
[16].龙景培,胡铭荣,王雄等.下肢深静脉血栓形成23例.医学理论与实践.2000;13(1):35
[17].Greenfield LJ.Current status of surgical therapy for deep vein thrombosis.Am J Surg 1985;10(suppl):64
[18].汪忠镐.重视血管外科的基本原则.中华实用外科杂志 2000;20(6):323
[19].金星,秦红松,尚德俊,中西医结合治疗下肢深静脉血栓形成206例临床观察,中国中西医结合杂志,1997,17(5):267
[20].高德,唐祖宣.《伤寒论》诊疗程序临床验证105例小结,1995,11(3):31-32
[21].王景春,王志平.辨治髂股静脉血栓形成210例.辽宁中医杂志,1996,23(6):265-265
[22].李玉幸,王凤莲.活血化瘀法治疗下肢深静脉血栓形成134例临床及实验观察.1997;19(1):13-14
[23].尚德俊,张秀英.中西医结合治疗下肢深静脉血栓形成研究进展.山东中医药大学学报.2000,24(4):309-313
[24].崔书克,刘成藏.中西医结合治疗下肢深静脉血栓形成120例[J].河南中医,2002,22(2):43-44.
[25].史以菊,吴俊芳,夏作理等.三七总皂甙对大鼠局灶性脑缺血的保护作用.中国临床药理学与治疗学杂志,1999,4(4):272
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