Objective:To establish the quality control method for compound Jieyu granules. Methods:Liquorice was identified by TLC. saikosaponin a, Saikosaponin d and rutin were determined by HPLC. Results:The spots on TLC plates were clear without any in-terference. The linearity was achieved within the range of 0. 508-16. 200 μg (r=0. 999 8) for saikosaponin a, 0. 503-16. 100 μg(r=0.999 7) for saikosaponin d, and 0.130-4.250 μg(r =0.999 9) for rutin. The average recovery was 99.7%(RSD =2.03%), 99. 8%(RSD=1. 44%) and 102. 6%(RSD=1. 40%), respectively. Conclusion:The method is simple, reliable and accurate, and can be applied as the quality control method for compound Jieyu granules.

Objective To investigate the effect of exosomes derived from hepatocellular carcinoma cells on the polarization of tumor-associated macrophages (TAMs), and to reveal the novel mechanism of hepatocellular carcinoma formation. Methods Hepatocellular carcinoma cell-derived exosomes were isolated by ultracentrifugation, and the characteristics of exosomes were identified by transmission electron microscope (TEM), Dynamic Light Scattering (DLS), and Western blotting. The model of macrophage polarization was induced and verified by quantitative real-time PCR and Western blotting. The t -test was used for comparison of normally distributed continuous data between two groups. A one-way analysis of variance was used for comparison between multiple groups, and the LSD- t -test was used for further comparison between two groups. Results TEM showed that hepatocellular carcinoma cell-derived exosomes were round or oval vesicles, LDS showed that the exosomes had a particle size of 172.65±2.34 nm, and Western blotting showed highly positive expression of the biomarkers TSG101 and CD63 in exosomes. There was a significant increase in the expression of CD68 after the addition of 15 ng phorbol ester to induce human-derived mononuclear macrophages for 24 hours to achieve adherent growth (1.00±0.25 vs 6.67±0.98, t =11.20, P < 0.001). Western blotting showed that compared with the control group (L02 cell-derived exosomes), the hepatocellular carcinoma cell-derived exosomes (at low, middle, and high doses) induced M2 polarization of macrophages and increased the expression of the markers Arg-1 and CD163 (all P < 0.05). Conclusion Hepatocellular carcinoma cell-derived exosomes promote M2 polarization of TAMs.