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Ultrathin sections (50 nm) were placed onto 200 mesh copper grids and double-stained with uranyl acetate and lead citrate before transmission electron microscopy analysis (HITACHI HT7700 Exalens)

Although the frequency of each cell population was not different between the declining and prolonged groups, analysis of the differentially expressed gene of each cluster revealed that the IL-1 expression levels of the CD14 + CD16 - classical monocyte cluster were significantly higher in the declining group than those in the prolonged group

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