PMC7499122_01
Preoperative adrenal CT and post-operative specimen of the patient. The right adrenal nodule with low density (arrow) was identified on CT (a), and surgical resection specimen revealed three nodules (red arrow) (b).
fendo-11-00593-g0001.jpg
PMC7499122_01_fendo-11-00593-g0001.jpg
F1
Adrenal computed tomography (CT) was performed, and a right adrenal nodule (13 mm in diameter) with low density was identified (Figure 1). Three nodules were found in the right adrenal resection specimen, and only the largest nodule was observed by CT (Figure 1).
Expression of CYP11B1/2 in three nodules of patients. Histological and immunohistochemical staining of three nodules (N1, nodule 1; N2, nodule 2; N3, nodule 3) of this patient and cortisol-producing adenoma from another patient (control, as a reference for positive CYP11B2 staining). HE, hematoxylin-eosin staining (a-d); CYP11B2: immunohistochemical staining of aldosterone synthase (i-l); CYP11B1: immunohistochemical staining of 11beta-hydroxylase (e-h); 200x magnification.
fendo-11-00593-g0002.jpg
PMC7499122_01_fendo-11-00593-g0002.jpg
F2
Immunohistochemical staining for the enzymes involved in aldosterone (CYP11B2) and cortisol (CYP11B1) (Figure 2) biosynthesis suggested that all the three nodules secreted aldosterone, whereas none of them secreted cortisol.
Gene sequence of KCNJ5 and PRKACA in three nodules. Genomic DNA was isolated from the patient's three adrenal nodules and amplified by PCR. Sanger sequencing detected a p.Leu168Arg mutation of KCNJ5 gene (A) in two nodules (N1, N3) and no p.Leu206Arg mutation of PRKACA gene in all three nodules. Positive control: a cortisol-producing adenoma harboring a p.Leu206Arg mutation of PRKACA gene (B); WT, wild type.
fendo-11-00593-g0003.jpg
PMC7499122_01_fendo-11-00593-g0003.jpg
F3
KCNJ5 mutations were found in two of the three nodules, whereas no PRKACA mutations were found (Figure 3).