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For pharmaceutical treatment, rapamycin (1.5 mg/kg/day) and PD98059 (10 mg/kg/day) were administrated to allogeneic recipients via intraperitoneal injection, respectively. Control animals received the appropriate vehicle injections. Treatment started on day 11 after surgery and lasted for 3 weeks. Two and 8 weeks after transplantation, the aortic grafts were harvested. Serial cross-sections (5 ��m thick) were cut. Six cross-sections from each animal were selected at 250-��m intervals for hematoxylin-eosin and Masson staining, followed by morphometric analysis. http://www.selleckchem.com/products/Rapamycin.html Immunohistochemistry and immunofluorescence were performed on cultured VSMCs and MSCs, as well as paraffin-embedded sections of arteries, using the avidin�Cbiotin peroxidase complex method. Apoptosis of VSMCs was detected using an In Situ Cell Death Detection Kit (Roche Diagnostics, Mannheim, Germany). Primary VSMCs were isolated from the descending thoracic aorta of 4-week-old Sprague-Dawley rats, using an explant http://www.selleck.cn/products/AP24534.html method (28). All experiments were conducted on cells between passages 3 and 8. Rat BM MSCs were obtained from femurs of Sprague-Dawley rats as previously described with some modifications (29). VSMCs were incubated with Opti-MEM I medium (Invitrogen) containing 200 multiplicity of infection (MOI) per cell of Ltv-vector or Ltv-p53 in the presence of polybrene (5 ��g/ml). Conditioned media (CM) from Ltv-p53-infected VSMCs (Ltv-p53 CM) or Ltv-vector-infected VSMCs (Ltv-vector CM) were prepared. Conditioned medium from uninfected http://www.selleckchem.com/products/jq1.html VSMCs (uninfected CM) was prepared as a control. MSC migration was performed using transwell chambers containing 5-��m pore filters (Corning, Inc., New York, NY, USA). MSC proliferation was quantified using a BrdU Labeling and Detection Kit (Roche Applied Science, Penzgerg, Germany). RNA was isolated from cultured SMCs or rat aortic grafts using TRIzol Reagent (Invitrogen, Carlsbad, CA, USA). RNA expression was analyzed by RT-PCR using iQ SYBR Green Supermix in an iCycler Real-Time PCR Detection System (Bio-Rad, Munchen, Germany). Western blotting was used to detect the expression of p53, TGF-��, PDGF-BB and SDF-1�� in cultured VSMCs, as well as the expression of signal molecules in cultured MSCs. All data were presented as mean �� SEM. After demonstration of homogeneity of variance with Bartlett test, one-way ANOVA, followed by Student�CNewman�CKeuls test where appropriate, was used to evaluate the statistical significance. Values of p