Detecting D-Dimer,APA,TAT By ELISA To Explore The Molecule Mechanism Of Osteoking In Rabbit TDVT Treatment

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 samplesAnimal 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(B₁,C₁ group,respectively 8),post-modeling 72h group(B₂,C₂ group,respectively 8) and post-modeling 120h group(B₃,C₃ 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 B₁ group,while the rabbits were selected randomly from group B to establish B₁ 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 B₂,C₂,B₃,C₃ respectively,then obtain blood and vascular specimen for the same method of group B₁.Statistical analysis about TDVT incidence rate:Information on the data in each group using x² 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 systemELISA were used to detect the blood D-dimer,APA,TAT expression of rabbits in A,B₁,B₂,B₃,C₁,C₂,C₃ 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.Results1.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 B₃ almost resolution.Group B₃ 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.