Figure 1.

Effect of SMI on tumor vessels in LoVo colon cancer xenograft mice. (A) Schematic plans of animal experiments for synergic anti-tumor effect study. Mice were intraperitoneally injected with SMI (10 mL/kg/day) and 5FU (15 mg/kg/3 days) for 14 days. (B) Images and volumes of tumors dissected from mice with 14-day treatment (n = 6). (C) Schematic plans of animal experiments for anti-angiogenic effect study. Mice were intraperitoneally injected with SMI for 15 days. (D) Morphologies of tumor vessels captured using multi-photon laser scanning microscopy (Zeiss) after 13 days of SMI treatment. Disordered vessels and branch points are indicated by white and yellow arrows, respectively. The color represents the depth of the vessel (depth = 180–250 μm). Statistical quantification of microvessels (<10 μm in vessel diameter) and branches were in the right side of the graph (n = 3–4). (E) Maximum intensity projection of the Z-positions with CD31 staining of tumor vascular endothelial cells. Disordered vessels and branch points are indicated by white and yellow arrows, respectively. Statistical quantification of CD31⁺ microvessels (<10 μm in vessel diameter) and vessel branches were shown in the right side of the graph (n = 3; thickness = 100 μm). (F) Double staining of CD31 (red) and α-SMA (green) of tumor slides (thickness: upper, 12 μm; lower, 100 μm). Vessels with pericyte coverage are indicated by white arrows. Statistical quantification of pericyte coverage based on calculations of the α-SMA⁺ fractions was shown in the right side of the graph (n = 3; thickness = 12 μm). *P < 0.05, ^**P < 0.01 vs. 5FU group for B. Data were expressed as mean ± SEM. *P < 0.05, ^**P < 0.01 vs. vehicle group for D, E and F. Scale bar = 100 μm. SMI: Shenmai injection.