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The brains were demineralized by EDTA and embedded in paraffin followed by 4% cold paraformaldehyde overnight for fixing. Specifically, 20 μm-thick sections were successively permeated, rinsed and differentiated based on standard procedures and well-prepared for microscopic examinations. The neurons and Nissl bodies in hippocampal CA1 regions were photographed using Olympus BX53 microscope (Olympus Co., Tokyo, Japan). Image-pro Plus 6.0 (Media Cybernetic Inc., Silver Springs, MD, USA) was used to calculate neuronal cellular number.
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