Endoscopic mucosal biopsies were obtained from the esophagus, gastric antrum and body, duodenum, terminal ileum, cecum, ascending, transverse, descending, and sigmoid colon, and rectum, respectively. Biopsy tissues were immediately fixed in formalin and processed in paraffin wax. Sections were cut at 3 μm and stained with hematoxylin and eosin for histopathologic examination.
Eosinophils were counted in five randomly selected high power fields (HPF). Quantification of eosinophils was performed using an Axioskop40 microscope (Mirax-Carl Zeiss, Oberkochen, Germany) at 400× magnification. Cell counting was performed by two pathologists who were blinded to the patients' statuses, and the average value over the five HPFs was calculated for each subject.
The histopathologic diagnosis of EoGID was made when the total number of infiltrating eosinophils per HPF exceeded 15 in the esophagus, 20 in the stomach and duodenum, and 25 in the colon and rectum. Terminal ileum and cecum were excluded, considering that tissue eosinophils may be present in normal children.20 (link)21 (link)22 (link)23 (link)