Kidney and heart specimens (4 mm) fixed in 10% formalin, paraffin-embedded, and stained with Masson's trichrome and Periodic acid-Schiff (PAS) reagent (Sigma-Aldrich). Semi-qualitiative histologic changes were assessed by a masked observer. The total numbers of kidney glomeruli were counted, and the degree of glomerular damage was estimated at 400× magnification from the degree of mesangial expansion in PAS stained tissue as follow: <25%, 25–50%, 50–80%, >80% [79 (link)]. Kidney interstitial fibrosis was estimated at 200× magnification on Masson's trichrome-stained sections using 10 randomly selected fields for each animal by the following semiquantitative criteria: 0, area of damage <5%; 1, areas of damage 5–10%; 2, damage involving 10–25%; 3, damage involving 25–50%; 4, >50% of the area being affected [73 (link)]. The cardiac fibrosis area was determined at 200× magnification in Masson's trichrome stained sections using ImageJ 1.36b (National Institutes of Health, USA).