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Thermal cycling consisted of initial activation (95C for 7 min) followed by 35 cycles of denaturation (95C, 30 s), annealing (56C, 30 s), and extension (72C, 30 s) and a final extention at 72C for 7 min

In this study, we choose the following immune cell marker genes as the label for each cell cluster and use the UMAP feature map and bubble map to show the expression of the marker genes 22 : Naive T (LEF1, CCR7), Effector memory CD8 + T (CD8A), Activated CD4 + T (IL7R), Circulating NKs (FCGR3A, TYROBP), Naive B cell (IGHD, TCL1A), Regulatory T cell (IL2RA), Memory B cell (AIM2, TNFRSF13B), NKs (GZMB, XCL1), Plasma cell (JCHAIN, MZB1), Monocyte (LYZ, S100A9), Proliferating CD4 + T (MKI67, TOP2A), Megakaryocyte (PPBP, PF4), DCs (PLD4, SOX4)

Pioglitazone carries an FDA warning about bladder cancer risk based on epidemiological signals, though absolute risk increases appear small

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