Preparation of cells
Using the Fresenius Kabi system, PBMCs were collected from each patient under electrocardiographic monitoring. The cells were cultured overnight, then adherent cells (including monocytes) and suspension cells (including lymphocytes) were collected separately.

Preparation of the DC vaccine
Isolated PBMCs were resuspended in a serum-free culture medium. The PBMCs were then seeded into a 175-cm3 flask to a concentration of 2-4×106/ml, divided among three bottles and then placed in a 37 ̊C, 5% CO2 incubator. After 2 hours of incubation, the 175-cm3 flasks were gently agitated; the suspension cells were discarded (or collected and used for culturing CIK cells), then washed 1-2 times with serum-free medium and, following addition of the DC medium, placed again in the 37 ̊C, 5% CO2 incubator. Autologous serum, granulocyte-macrophage colony-stimulating factor, interleukin (IL)-4 and tumor necrosis factors (all from PeproTech EC, Ltd., London, UK) were added on the 3rd day. The tumor antigen was added on the 5th day and the ripening factor was added following addition of the tumor antigen. DC cells were collected between the 7th and 8th days and then washed three times with saline.

Preparation of CIK cells
The culture bottles were pre-coated with a 5 μg/ml final concentration of CIK reagents before preparing the CIK cells, then placed in a 4 ̊C environment overnight and washed with phosphate- buffered saline 2-3 times on day 0. The CIK cells were resuspended in the initial CIK cell medium at a concentration of 2-3×106/ml and spread into a culture bag according to the number of cells. Autologous plasma was then added and cultured in a 37 ̊C, 5% CO2 incubator. IL-2, interferon 2γ and CD3 monoclonal antibody (100 μg/ul) were added using 5-ml disposable syringes with needles.