Before surgery, the

animals were kept under standard laboratory conditions. In brief, a 1.5 cm side-to-side surgical EGDA was created between the first duodenal loop and the gastro-esophageal junction, about 3 cm distal to Treitz’s ligament, with accurate mucosa-to-mucosa opposition (Figure 1), so that duodenal and gastric contents flowed back into the esophagus. Unlike other models, this “”Kumagai-Hattori”" model check details preserves the animal’s normal stomach function and nutritional status [19, 21, 22]. Figure 1 Pathology findings of the esophageal cancer model. (A) Schematic illustration of the surgical intervention of the Kumagai-Hattori model (left) and selleck chemicals llc representative macroscopic picture (right): unfixed esophagus, stomach and jejunum (excised en bloc) are opened through the dorsal wall (mucosal surface upward). (B-G) Histological findings observed (H&E staining): (B) anastomosis ulcer;

(C) squamous cell polypoid hyperplasia; (D) multilayered epithelium; (E) specialized columnar epithelium (intestinal metaplasia); (F) adenocarcinoma; (G) squamous cell cancer. (Original magnifications, 40×, 20× and 10×) Postoperatively, Bioactive Compound Library high throughput the animals had free access to water and food. No treatments with any known carcinogen were applied. Ten of the 74 rats died (mainly of respiratory complications) within 7 days after surgery and were not considered. As in already published experimental models, the animals were sacrificed

at different times after surgery (i.e. Group A [22 rats] after <10 weeks [range = 3–9.9], Group B [22 rats] after 10–30 weeks [range = 10–29.7], and Group C [20 rats] after >30 weeks [range = 31–54]) [19, 21, 22, 27, 28]. This study was approved by the Institutional Animal Care Committee of the University of Padova. All procedures were performed in accordance to the Italian law on the use of experimental animals (DL n. 116/92 art. 5) and according to the “”Guidelines on the Care and Use of Laboratory Animals”" (NIH publication 85–93, revised in 1985). Pathology Immediately after death, the thoracic and abdominal cavities were examined and the esophagus, stomach, and jejunum were excised en bloc. The esophagus was opened longitudinally through the dorsal wall. With Glutamate dehydrogenase the mucosal surface uppermost, the margins of the specimen were fixed to a cork plate with pins. Gross specimens were fixed in 10% neutral-buffered formalin for 24 hours. All specimens were examined grossly (see gross pathology) and cut serially (2–3 mm thick coronal sections). The tissue samples were routinely processed. Tissue sections 4 μm thick were obtained from paraffin blocks and stained with Haematoxylin & eosin. Lung, liver, kidney and spleen tissues were also collected for histological assessment. Two experienced gastrointestinal pathologists (GI & MF) reviewed all the slides.