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For cells, they were first washed with PBS and fixed at room temperature for 15 min, followed by incubation with the staining solution at pH 6.0 overnight at 37 C

Macroscopic Assessment The depth of destruction in articular cartilage was evaluated according to the measures described by Pelletier et al 32 using a four-point scale (0 = normal-appearing surface, 1 = negligible fibrillation or a minor yellowish discoloration of the surface, 2 = destruction spread out into the superficial or central layers only, 3 = destruction that spread into the deep layers, and 4 = destruction that spread to the subchondral bone)

180 NockNLBockCNeslund-DudasCBeebe-DimmerJRundleATangDet al

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Monozygotic female twins discordant for phenotype of Wilsons disease