Adult male Sprague-Dawley rats were injected with either LENK-SQ-Am NPs (20 mg/kg) or 5% dextrose (n = 3 animals per group). At 24 or 48 hours after injection, the animals were anesthetized with 2% isoflurane gas in oxygen flow (1 liter/min), and blood was collected from tail blood vessels by anticoagulant (heparin, 500 UI/ml)–treated syringes. The blood samples were centrifuged at 1000g for 15 min, and the plasma was collected and stored at −20°C before analysis. The AST and ALT levels in plasma were analyzed by Cerba Vet, France.

In a separate experiment, 24 or 48 hours after injection, rats were deeply anesthetized by Dolethal. Liver, kidneys, spleen, heart, and lungs were then excised, fixed by 4% formaldehyde, paraffin-embedded, and cut into 5-μm-thick sections. Hematoxylin and eosin staining was performed on all the organs for analysis of the morphology (Zeiss).

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