Objective To elucidate the immunologic mechanism and clinical effect of high intensity focused ultrasound (HIFU) combined with dendritic cell and cytokine induced killer cell (DC-CIK) immunotherapy on patients with pancreatic cancer.Methods Seventy-two pancreatic cancer patients were divided randomly into 2 equal groups,one treated with HIFU only the other treated with HIFU and DC-CIK immunotherapy.Ultrasound imaging and a variety of immunological indexes were recorded before and after treatment and the clinical effects in the two groups were compared.Moreover,autogenous tumor cells were isolated from the combination therapy group and the killing activity of DC-CIK which loaded tumor antigen processed by HIFU on autogenous tumor cells was observed.Results Tumor antigen processed by HIFU can improve the killing activity of DC-CIK on autogenous tumor cells.After treatment,the immunological indexes,of all patients were better than before treatment.(58.26 ± 17.97 versus 52.15 ± 14.22 pg/ml with IL-12 22.14 ± 6.39 versus 17.36 ± 5.73 ng/ml with HSP70 and 0.94 ± O.34 versus 1.32 ± O.61 ng/ml with TGF-β,P ＜ 0.05 ) ; The combination group was significantly better than the HIFU group with regard to the average scores of quality of life (75.89 ± 19.65 versus 67.22 ± 16.34,P＜0.05),pain (3.15 ±0.82 versus 3.59 ± 1.04,P ＜0.05),tumor markers (107.55 ±27.58 versus 123.63 ±34.12 U/ml) and survival time (18.92±6.47 versus 13.36 ±5.78 mos).Conclusion HIFU can improve the immunologic status and anti-tumor response in patients with pancreatic cancer.HIFU combined with DC-CIK has good synergistic therapeutic effect for treating pancreatic cancer.

BACKGROUND: As a kind of semitransparent membrane, human amniotic membrance contains many kinds of nutrients, which is a good biological material loaded with keratinocytes.OBJECTIVE: To construct epidermal substitute of the skin from human amniotic membrane loaded with porcine keratinocytes and examine the morphological characteristics of the growth and proliferation of keratinocytes seeded on human amniotic membrane.DESIGN: Single sample study and repetitive measured observation based on the cells.SETTING: Institute of Combined Injuries of Chinese PLA, Academy of Preventive Medicine, Third Military Medical University of Chinese PLA.MATERIALS: The experiment was completed in the State Key Laboratory of Trauma, Burn and Combined Injury and Institute of Combined Injuries of Chinese PLA, Academy of Preventive Medicine, Third Military Medical University of Chinese PLA from January to November 2001. Porcine keratinocytes was collected from Guizhou minipigs aged 3 weeks.METHODS: The primarily cultured keratinocytes of Guizhou minipigs were subcuhured, expanded and bred on the stroma surface of human amniotic membrance at the density of 1.63 × 105/cm2. The growth and proliferation of keratinocytes were observed under inverted microscope every day. From the 3rd day and the 15th day after being cultured, the growth of keratinocytes on human amniotic membrane was examined under light microscope and electron microscope.MAIN OUTCOME MEASURES: The growth of keratinocytes on human amniotic membrane was examined RESULTS: Keratinocytes evidently adhered to the stroma surface of human amniotic membrane about 30 minutes after being cultured, which was observed under inverted microscope. Most keratinocytes grew and adhered to the stroma surface of human amniotic membrane within 24 hours. Monolayer of keratinocytes formed and completely covered human amniotic membrane within 3 days. It was observed under the light microscope that the monolayer of keratinocytes adhered to human amniotic membrane and arrayed tightly. The keratinocytes presented in the shape of polygon, and plasmalemmas of keratinocytes formed many pseudopods under the observation with scanning electron microscope. Keratinocytes adhered to human amniotic membrane well and with many keratinofilaments in them under the observation with transmission electron microscope. Keratinocytes arrayed on human amniotic membrane densely with many cellular debris and some keratinocytes formed cavitations in them due to aging after growth for 15 days under the observation with inverted microscope.CONCLUSION: Human amitotic membrane is a good carrier of keratinocytes cultured on it in vitro, and is able to promote the proliferation of keratinocytes significantly. However, when keratinocytes were loaded on the human amniotic membrane for 15 days, some keratinocytes formed cavitations in them due to aging.