Pharmacodynamic analysis
Collection of peripheral blood mononuclear cells (PBMCs) were extracted from whole blood in a CPT Vacutainer as recommended by the manufacturer. Briefly, 8 ml of whole blood were collected from patients at baseline, week 12 and 28 and centrifuged at 1500 × g for 20 min at room temperature to isolate the PBMC fraction. The PBMC fraction was then transferred to a 15-ml conical tube, with phosphate buffered saline (PBS) and added PBS to fill the tube and were centrifuged at 600 × g for 10 min at room temperature. After PBS was aspirated, the PBMC pellet was snap frozen and stored at 80°C until use. For analysis, using western immunoblotting was used and probed with phospo-p70S6 kinase and GAPDH antibody (both purchased from Thermofisher).