Statistical Analysis Treatment groups were compared using analysis of var iance, paired check this t test or Wilcoxon signed rank test, depending on data distribution, using the JMP software. A p value of 0. 05 was considered statistically significant. All values were expressed as mean SEM. Statistical significances were denoted with three different symbols. Results The current experimental studies include three parts quality control of APS. effects of APS on mice. effects of APS on the apoptosis of cells. In part 2, mice were subjected to radiotherapy to create a bone marrow damaged model. Then we study the effects of APS on the recovery of various hemato poietic, especially those thrombopoietic cells. All treat ments were conducted on these control mice. In part 3, apoptosis were induced in cells by serum depletion.
The effects of APS were tested on these apoptotic cells. In our experiments, TPO was used as the positive control. And a PI3K inhibitor was used to test if APSs effects implicate the PI3K pathway. Quality control of APS Since the APS preparations usually contain various che mical compositions, qualitative and quantitative defining the experimental materials is important. This is to ensure that various preparations of RAS have consistent biological activities and any follow up studies can repro duce the results generated in previous studies. Here, we used two methods to define our APS preparations. First, the RAS samples were subjected to UV spectral analysis and the spectrum were compared to that of a standard. The IR spectrums of our RAS experimental material and the RAS standard material are shown in Figure 1.
The above analyses were repeated five times and the correlation coefficients of the spectrums were found to be greater than 0. 9973. Then we mea sured the total amounts of polysaccharides in the APS preparations using the anthrone sulfuric acid assay. Cilengitide Glu cose was used as the standard and the standard curve was found to be y 0. 0069 x 0. 0045 with a correla tion coefficient of 0. 9992. By comparing to the standard curve, polysaccharides were found to represent 90% of the total APS preparations. Both bacterial endotoxins or lipopolysaccharide and b D glucan can trigger TAL gelation, which is used for the detection and semi quantification of the LPS or b glucan content of the APS preparation. We found that the solid gel was formed for APS dilutions at concentrations of 5, 1. 667, 0. 556, and 0. 185 mg/ml. In comparison, gelation occurred at concentrations of 0. 125 and 0. 06 EU/ml for the control standard endo toxin. So the LPS or b glucan content of the APS pre paration was estimated to be approximately 0. 06/0. 185 or 0. 32 EU/mg of APS extracts. This is equivalent to 0. 128 ng endotoxin in 1 mg of APS.