Differential gene expression analysis for own control of single subject

For HSCR segments, the normal segment of each subject serves as his or her own control of the aganglionic segment, allowing sufficient power to detect a significant effect of the segment with only a small number of subjects. The genes that appeared in the three or above subjects were the potential DEGs. The number of myofibroblasts and smooth muscle cells was too few to perform the differential expression analysis in several subjects, so we merged the two cell types for further analysis.