Hippocampal transcriptomics of PND group and control group
12 PND models were established by exploratory laparotomy under 1.4% isoflurane anesthesia as reported previously. Briefly, mice were anesthetized by the inhalation of 2.0% isoflurane (100% oxygen flow 1L/min), and after the mouse righting reflex disappeared, 1.4% isoflurane anesthesia was maintained (100% oxygen flow 1L/min)[20]. After skin preparation and disinfection, stratified incision was made along the median abdominal line to make a 2.5cm longitudinal incision. The left and right rectus ophthalmic forceps were separated and a mesangial arterial loop (3-5cm) was gently pulled out of the ileum. The ileum (3-5cm) was placed on the surface of normal saline wet sterile gauze and the exposed intestinal segment was covered to protect the incision. After the intestinal contents were removed, the exposed intestinal segment was evenly and gently twisted with the thumb, indicator finger, and middle finger for around 10 min, separated by gauze. Be careful to knead the intestinal segment gently to avoid mesenteric arteriovenous bleeding or serious mechanical tissue damage. The small intestine was then reinserted and the incision closed with 4-0 absorbable sutures. Finally, 5% lidocaine cream was used for analgesia after surgery. The procedure was conducted under sterile conditions and lasted approximately half an hour on a thermal blanket. The 12 control models did not receive any treatment. After RNA extraction and library construction, Illumina NovaSeq 6000 and Fastp were used for RNA sequencing and assessing the quality of the sequencing data; then, clean reads were consistent with the mouse genome (GRCm38/mm10) using Hisat2. The reads number of each gene was counted by RSEM, and the FPKM of each gene was calculated.