After a 24-h period of fixation in 4% paraformaldehyde, the intestinal segments were taken out, and dehydrated using the graded concentrations of ethanol (70% to 100%) and chloroform56. After dehydration, the segments were embedded in paraffin, and then placed in a refrigerator to make the paraffin sufficiently hard. Cross-sections of the segments were cut at approximately 5 µm thickness with a microtome and stained with haematoxylin and eosin56. In each section, 10 fields were examined using a light microscope with a computer-assisted morphometric system (BioScan Optimetric, BioScan Inc., Edmonds, WA). The villous height (VH), the associated crypt depth (CD), and villous width were measured, and then the ratio of villus height to crypt depth (VH/CD) and villous surface area were calculated. Villus height is defined as the distance from the villus tip to crypt mouth, and crypt depth is defined as the depth of the distance from the crypt mouth to the base, and villus width is defined as the width at half of villus height59. All intestinal histological analyses were done by the same person, who was blinded to the treatments.

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