Bcl-2, Bax and caspase-3. A: Bcl-2 expression. Compared with sham animals, vehicle treated group showed a slight more positive staining of Bcl-2 cells. Hydrogen saline increased the number of Bcl-2 cells (Upper panel: 100 ×; Lower panel: 200 ×; Scale bar: 20 μm). B: Bax expression. A marked increase of Bax positive cells was observed in samples collected from vehicle treated animals, and hydrogen saline reduced the number of Bax positive cells (Upper panel: 100 ×; Lower panel: 200 ×; Scale bar: 20 μm). C. Hydrogen saline applied at 3 h after reperfusion increased the number of positive BCl-2 cells and reduced the number of positive Bas cells at 24 h after MCAO (**P< 0.01 vs. MCAO). D. A marked increase of caspase-3 activity was observed in the vehicle treated animals and hydrogen saline applied at 3 h after reperfusion decreased caspase-3 activity at 24 h after MCAO (**P< 0.01 vs. MCAO).