Therefore, the sera from healthy volunteers and patients with NAFL, NASH, and NASH-HCC were fractionated by size-exclusion chromatography, and the fractionated samples were assayed for IgM-bound AIM and IgM-free AIM by ECLIA using beads coated with anti-AIM antibody No. 11 and Ru-labeled anti-AIM antibody No. 8. This evidence concerns the gene CD40LG and hepatocellular carcinoma.