Histopathological changes in liver tissues with AHF were examined by H&E staining according to our laboratory methods previously described13. Briefly, approximately 5×5×3-mm cuboids from the right lobe of the liver were routinely fixed, washed, dehydrated, cleared and embedded in paraffin. Afterwards, liver sections (4 μm) were stained with H&E (Beyotime Institute of Biotechnology) according to the manufacturer’s instructions. The liver sections were dehydrated with gradient ethanol, cleared in xylene and sealed with neutral gum, and then histological assessment was performed under a microscope.