| Objectives:1To make further study on outcomes as hemorheology, erythrocyte (RBC), platelet (PLT), prothrombin time (PT), etc. for blood stasis syndrome with early formation of post-trauma stasis at early stage after trauma (injury) and explore intervention effect and mechanism of Astragalus membranaceuss for such pathological stage.II To exploratorily apply with principal component analysis (PCA) and analysis of variance (ANOVA) for data study and observation from both overall and partial perspectives, and find information on changes within outcomes and correlations among outcomes during early post-injury blood stasis to provide future research on syndromes and conditions according to Traditional Chinese Medicine (TCM) with new consideration.Methods:210rabbits were randomized into blank controlled (BC) group, modeling group and TCM group. Then such3groups were divided into7groups as1hour (H) group,8H group,24H group,48H group,72H group,120H group and168H group. The post-injury blood stasis model of rabbit was set by quantitative striking. While other2groups were offered normal saline (NS), TCM group were fed with decoction. Blood samples were collected at certain times for examination of17outcomes as hemorheology, RBC, PLT, PT, etc selected and determined based on literature. PCA was applied with dimensionless and dimension reduction process for outcomes examined, and several un-related principal components were set. Then new variances as principal component scores were calculated with one-way ANOVA. Hence results were interpreted with contribution rates besides correlation coefficients of each principal component. Outcome-groups and single outcomes in each group were analyzed to make conclusions. Results:I Results for modeling groupThere were5principal components with eigenvectors over1generated from PCA among17outcomes were selected in modeling group, with accumulative contribution rate as85.09%. The contribution rate of the first principal component was achieved by26.87%, including information on outcomes (indicators) as RBC, whole blood viscosity (at low-shear, medium-shear and high-shear rates) and plasma viscosity. Such outcomes increased at1H, then decreased at120H and168H significantly. Contribution rate of the second principal component produced22.226%, which contained PT, PLT, platelet distribution width (PDW), mean platelet volume (MPV) and platelet-large contrast ratio (P-LCR). Such indicators showed no change at7time-points. Contribution rate of the third principal component presented13.69%, expressing indicators as mean corpuscular-hemoglobin concentration (MCHC), red blood cell distribution width (RDW) and coefficient of variation for red blood cell volume (RDW-CV). MCHC increased within1H, meanwhile values of RDW and RDW-CV decreased. No other changes were found at other time-points. The fourth principal component contributed12.61%with hemoglobin (HGB) and hematocrit (HCT). Both increased at1H,8H and168H while declined at48H and120H remarkably. The fifth principal component covered both mean corpuscular volume (MCV) and mean corpuscular hemoglobin (MCH) with contribution rate as9.70%. Both outcomes increased at1H and120H and declined at72H and168H obviously.II Results for TCM groupSix principal components selected by eigenvectors more than1generated from PCA among17outcomes were studied for TCM group, producing total accumulative contribution rate as85.09%. The first principal component contributed22.20%. RBC, HGB, HCT, RDW and RDW-CV were included, and the former three outcomes increased after1H, and fell at120H. Whereas latter two outcomes fell at1H yet increased at72H and120H. The second principal component offered21.73%portion of contribution, and outcomes as PDW, MPV, P-LCR, PLT as well as plasma viscosity were clustered in this part. The former three ones dropped followed with raised at1H, remained above normal level at72H and168H, and fell at1H and120H. Compared with that, the latter two outcomes raised followed by dropped at1H and then decreased, reduced at72H and168H, and increased at120H. The third principal component contributed for15.21%. Whole blood viscosities at different shear rates (as low-shear, medium-shear and high-shear) were grouped in this part. Such indicators rose at1H and reduced at8H. The fourth principal component made11.53%of total contribution. MCV and MCH were studied. Both increased at24H. The fifth principal component reported9.51%. MCHC was focused and stated no significant change at any time-point observed. As for the sixth principal component, which contributed for6.03%. This part was related with PT. The outcome dropped at72H significantly and increased at120H and168H.Ⅲ Analysis for Time-point Resultsi1H results:5principal components generated in1H group, offering accumulative contribution rate of86.21%in total. Low-shear viscosity, high-shear viscosity, plasma viscosity, MCV, RDW-CV, PLT, PDW, MPV as well as P-LCR were included in the first principal component (contribution rate:30.29%), followed as RBC, HCT, MCH and MCHC in the second principal component (20.96%), HGB and PDW in the third principal component (14.25%), medium-shear viscosity in the fourth principal component (12.76%), and PT in the fifth principal component (7.95%).Change in outcomes:Outcomes as low-shear/medium-shear/high-shear viscosity, plasma viscosity, MCV, RDW-CV and PLT increased at this time-point, but Astragalus membranaceuss made these values decrease. PDW, MPV and P-LCR decreased yet were promoted ascending in values by fed of Astragalus membranaceuss decoction. No change found in PT at1H after trauma (injury), and Astragalus membranaceuss shortened this period in our study. There were no significant changes observed in other outcomes among BC group, modeling group and TCM group. ii8H results:Five principal components were generated in this group (85.71%). The first principal component (30.93%) contained PLT, HCT, MCV, PDW, MPV besides P-LCR. The second principal component (18.90%) covered RBC, HGB, MCHC, RDW and RDW-CV. Three outcomes were classified in the third principal component (13.58%), such as PT, low-shear viscosity plus high-shear viscosity. Only medium-shear viscosity in the fourth principal component (12.23%), leaving MCH in the fifth principal component (10.07%).Outcome changes:PT and whole blood viscosities (low-shear, high-shear) didn’t showed obvious change at8H. Astragalus membranaceuss reduced these outcomes. No significant change was detected in other outcomes in all3groups.iii24H results:Six principal components in this group (88.323%). The first principal component (22.50%) were comprised by MCHC, PDW, MPV, P-LCR and PLT. The second principal component (20.71%) such as high-shear viscosity, RBC, HGB and HCT. The third principal component (14.91%) as plasma viscosity, RDW-CV and MCH. The fourth principal component (12.19%) as low-shear viscosity and medium-shear viscosity. The fifth principal component (11.52%) as MCV. The sixth principal component (6.50%) as PT.Outcome changes:MCHC, PDW, MPV, P-LCR, PLT, RDW-CV, MCH besides plasma viscosity presented no significant change at24H. Astragalus membranaceuss made both PLT and MCH increased, while led decrease in the other6outcomes. Whole blood viscosities at low-shear and medium-shear rate went down at24H, and Astragalus membranaceuss increased such dropping tendency. No remarkable change found in other outcomes among three groups.iv48H results:Six principal components in this group (85.50%). The first principal component (28.99%) plasma viscosity, HGB, HCT, RDW, PLT, PDW, MPV and P-LCR. The second principal component (17.69%) such as high-shear viscosity, RDW-CV, MCV and MCH. The third principal component (13.50%) as PT and RBC. The fourth principal component (9.64%) as MCHC. The fifth principal component (9.03%) as low-shear viscosity and medium-shear viscosity. However, the sixth principal component (6.65%) indicated not clear information concerning any outcome.Outcome changes:17outcomes in all three groups showed not significant change at post-trauma48H.v72H results:Six principal components in this group (85.99%). The first principal component (29.76%) reported MCV, MCH, RDW, PDW, MPV, P-LCR, plasma viscosity, RBC and PLT. The second principal component (18.88%) such as PT, HGB and HCT. The third principal component (12.68%) indicated no clear information concerning any outcome. The fourth principal component (10.327%) as RDW-CV. The fifth principal component (8.18%) as low, medium and high-shear viscosity. Moreover, the sixth principal component (6.16%) as MCHC.Outcome changes:There were no significant change in MCV, MCH, RDW, PDW, MPV, P-LCR, RBC, PLT, MCHC or plasma viscosity at post-trauma72H. Astragalus membranaceuss reduced levels in RBC, PLT, MCHC plus plasma viscosity while increased MCV, MCH, RDW, PDW, MPV and P-LCR. Furthermore, significant changes were not tested in other indicators among BC group, modeling group and TCM group.vi120H results:Six principal components in this group (87.52%). The first principal component (30.88%) reported RBC, HGB, HCT, MCV, RDW, RDW-CV and PLT. The second principal component (19.20%) such as PDW, MPV and P-LCR. The third principal component (12.64%) as medium-shear viscosity and MCH. The fourth principal component (11.17%) as PT, MCHC and high-shear rate. The fifth principal component (7.39%) as low-shear viscosity. The sixth principal component (6.25%) as plasma viscosity.Outcome changes:Three outcomes, RBC, HGB and HCT, decreased at post-trauma120H in both modeling group and TCM group. On the other hand, MCV, RDW, RDW-CV besides PLT increased significantly in both groups. Other outcomes reported no significant change among3groups.vii168H results:Five principal components in this group (85.19%). The first principal component (35.84%) reported low, medium and high-shear rate, PLT, plasma viscosity, HGB, HCT, MCV and MCH. The second principal component (20.69%) such as MCHC, PDW, MPV and P-LCR. The third principal component (12.19%) indicated information on RBC and RDW-CV. The fourth principal component (9.86%) as PT and RDW. The fifth principal component (6.61%) indicated unclear information about any outcome.Outcome changes:17outcomes were found unchanged in three groups at this time-point.Conclusions:Ⅰ There are significant changes in both thin fluid and thick fluid inside rabbit during early post-trauma stage. Thus this indicates delivery and distribution of thin and thick fluids as a vital pathological condition for this period.Ⅱ Astragalus membranaceuss may resist blood-stasis syndrome at early post-trauma stage through promotion of distribution of thin and thick fluids, which performs more obviously in1hour after trauma/injury.Ⅲ Common factors to induce change in outcomes could be found through overall study on outcome groups. Such method of comprehensive analysis on outcomes to pursue common performance system is worked according to conceptions of TCM syndromes. |
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