Furthermore, we established the ECLIA method capable of measuring both IgM-free AIM and IgM-bound AIM (total AIM); we also investigated the existing AIM form in blood by measuring the fractionated samples from the sera of patients with NASH and NASH-HCC using size-exclusion chromatography to investigate the clinical significance of IgM-free AIM in NASH-HCC. This evidence concerns the gene CD40LG and hepatocellular carcinoma.