Fig 2: Baseline aCECs levels of placebo group and anlotinib group. A, CD31-labeled aCECs levels of the two groups. B, CD105-labeled aCECs levels of the two groups. aCECs, activated circulating endothelial cells

Fig 3: Characterization of human umbilical cord mesenchymal stem cells (UC-MSCs). Isolated primary UC-MSCs were cultured to passage 5, and then characterized for their surface markers and multipotent differentiation capacity. A typical expression pattern of surface markers for mesenchymal stem cells (MSCs) was determined by flow cytometry with high positive expression of CD105, CD90, CD73 (> 98%), and negative expression of CD19, CD34, CD14, CD45, and human leucocyte antigen-DR (HLA-DR) (< 0.3% positive). Multiple differentiation capacity of UC-MSCs was determined with different induction media as described in Methods. The phenotypes of induced differentiation were visualized by staining with Oil Red O for adipogenesis (scale bars = 50 µm), Alizarin Red for osteogenesis, and Alcian blue for chondrogenesis (scale bars = 100 µm) as indicated, respectively. After induction, a significant enhanced expression of adipogenic, osteogenic, and chondrogenic genes was detected by quantitative real-time polymerase chain reaction (*p < 0.05, **p < 0.01 vs control). Adipoblast related genes were as follows: FABP4, fatty-acid-binding protein 4; PPAR?, peroxisome proliferation-activated receptor ?; C/EBP-a, CCAAT/enhancer-binding proteins a; Pref-1, preadipocyte factor 1; LPL, lipoprteinlipase. Osteoblast related genes were as follows: BGLAP, bone gamma carboxyglutamate protein; SP7, Sp7 transcription factor; Runx2, runt-related transcription factor-2; OPN, secreted phosphoprotein 1; ALP, alkaline phosphatase. Chondrocyte related genes were as follows: ACAN, aggrecan; MIA, melanoma inhibitory activity; COL1A2, collagen type I a 2 chain; COL2A1, collagen type II a 1 chain; COL10A1, collagen type X a 1 chain. mRNA, messenger RNA.

Fig 4: Analysis of PFS and OS in anlotinib group with a cutoff value of aCECs min/baseline = 1. A, PFS of CD31-labeled aCECs min/baseline <1 and aCECs min/baseline =1 group. B, OS of the two groups. C, PFS of CD105-labeled aCECs min/baseline <1 and aCECs min/baseline =1 group. D, OS of the two groups. PFS, progression-free survival; OS, overall survival. aCECs, activated circulating endothelial cells

Fig 5: The clonogenic capacity and surface marker expression of hFDSPCs. The clonogenic capacity of hFDSPCs was examined using CFU assays. a hFDSPCs cultured at P1 to P3 were seeded in 6-well plates at a density of 50 cells/cm2, with cell clones observed after 7 days. b Colony formation numbers were counted; the data were plotted in graphs and analysed using Graph Pad Prism Software (n = 6). c The colony morphology was observed under an inverted microscope, bar = 1000 µm. d Immunophenotyping of hFDSPCs was characterised using flow cytometry analysis. hFDSPCs were analysed for expression of the following markers (n = 4): CD90 (97.53% ± 0.48%), CD44 (97.28% ± 0.52%), CD105 (22.57% ± 2.09%), CD34 (1.30% ± 0.75%), CD45 (0.73% ± 0.39%), CD29 (98.62% ± 1.18%), CD13 (98.06% ±0.26%), CD59 (98.80% ± 1.02%), CD31(2.07% ± 0.31%), and CD133 (0.88% ± 0.16%). Isotype-matching IgG-FITC and IgG-PE were used to determine nonspecific signals. Data are shown as means ± SD, **p < 0.01, ***p < 0.001