Objective To investigate the method and optimum conditions of electric transfection,and the major influential factors of electransfection efficiency and the survival rate of dendritic cells. Methods RNA was extracted from human hepatocarcinoma cell line(Bel 7402).Purified monocytes as precursor DC-s(pDC-s) were separated from human peripheral blood cells(PBMCs) by density gradient centrifugalization with lymphocyte gradation fluid and adherence method,pDCs were incubated in RPMI-1640 medium containing rhGM-CSF(8?10~5IU/L) and rIL-4(5?10~5IU/L) for 7 days and made them fully differentiate into immature DCs(imDCs).The total RNA human hepatocarcinoma cell and green fluorescent protein(GFP) were electransfected respectively into imDCs by electroporation apparatus with different electric voltages,times of impulse,cell concentrations,temperatures and electroporation buffers.Numbers of green fluorescence positive cells and the total cell number were counted respectively under fluorescent microscope,and visible light microscope.One day after the electric transfection,the cells were stained with 0.4% trypan blue,and electransfection efficiency and the cell survival rate were counted. Results Electransfection efficiency was increased to the highest value,up to about 49.7% when imDCs with the concentration of 5?10~6 cells/ml were mixed with 40?g-total RNA of human hepatocarcinoma cell,the electric voltage of electroporation apparatus was set at 300V,and the time of impulse was 500Us.Conclusion Electric transfection provides a technical possibility to make human hepatocarcinoma RNA into imDCs.The major influential factors of the electransfection efficiency were electric voltage and impulsing time.As receptor cells,the imDCs growing condition was also an important influential factor.

Autoimmune pancreatitis (AIP), which is always associated with autoimmune manifestations, was introduced in 1995 and has been recognized as a type of chronic pancreatitis. Despite numerous studies in Japan, Europe and the United States in recent years, no consensus has been reached about the diagnostic criteria for AIP that may be difficult to distinguish from pancreatic cancer(PC). Nevertheless, the results find it dramatically responds to steroid therapy.

Objective To investigate the distribution and the number of splenic lymphocytes in the patients with severe acute respiratory syndrome(SRAS).Methods Immunohistochemical method with four specific antibodies(CD3,CD4,CD8,CD20) was used to detect the distribution of lymphocyte special antigens in the spleens of six cases who died of SARS and six normal cases as the controls.The numbers of lymphocytes were analyzed with image analysis system.Results In the spleen of SARS patients,splenic corpuscle and periarterial lymphatic sheath (white pulp) were damaged severely. In the number of periarterial lymphatic sheath decreased 93.39% and in the number of splenic capsule decreased about 80% even disappeared at all. Red pulp hemorrhage necrosis was widely spread. In red pulp the number of CD3~(+)T cells decreased 71.76%,even disappeared at all,in the number of CD4~(+) and CD8~(+)T cells markedly decreased 86% and 84% respectively.The number of CD20~+ B cell decreased more than 80%. Conclusion T cells and B cells in spleens of SARS patients decreased widely spread and markedly in number,indicating that the immune system had been damaged severely,and the destroyment of the immunosystem may be the primary lesion of SARS.

Objective To investigate the number,distribution and expression intensity of dendritic cells and T lymphocytes in human cervical intraepithelial neoplasia and invasive carcinoma for seeking biotherapeutic evidence of cervical carcinoma. Methods The number,distribution and expression intensity of CD-(1a),S-100,CD-3, CD-8 positive cells were determined in cervical intraepithelial neoplasia and invasive carcinoma and human normal cervix tissue as the control,by using immunohistochemical technique and computerized imaging analysis system. Results Compared with human normal cervical tissue,the number and the expression intensity of CD~+-(1a),S-100~+,CD~+-3,CD~+-8 cells in cervical intraepithelial neoplasia significantly increased(P

Objective To investigate the distribution and significance of CD20 positive cells in the early human embryo. Methods Distribution and morphology of CD20 positive cells in 10 specimens of human embryo aged from 6 to 7 weeks were brown,and studied with immunohistochemical method. Results 1.CD20 positive cells appeared in the liver of early human embryo.2.The immunohistochemical positive substances were brown,and found chiefly in the nuclei of B cells as unevenly-distributed granules,and were not detected in the cytoplasma and on the cell membrane.Conclusion The accuracy of location and distribution of CD20 positive cells in the nuclei of B cells of early human embryo liver may provide an important clue for further exploration of the functional mechanism of CD20 in the process of B cell proliferation and differentiation.

Objective To analyse the number of dendritic cells and macrophages and the size of macrophages in the spleen of SARS patients so as to provide evident for the study of pathology and pathogenesis of SARS. Methods Immunohistochemical method with four specific antibodies(S-100,CD68,HLA-DR,CD83) were used to detect the dendritic cells and macrophages in the spleens of six dead patients of SARS and six accidental deaths as the controls,The number or the size of these positive cells was analysed with image analysis system. Results In the spleens of SARS patients,the number of S-100~+ dendritic cells in the white pulp was reduced by 80.4% on average,and even disappeard.The number of CD68~+ macrophages in the red pulp was reduced by 39.48% in SARS spleens,and the average size of individual macrophages was increased by 2.21 times.HLA-DR~+ antigen presenting cells(APC) reduced remarkably in the SARS spleen white pulp.CD83~+ mature dendritic cells did not exist in either SARS spleens or the control spleens.Conclusion The function of antigen presentation had been damaged severely,which supports that SARS should be categorized as a viral immune deficiency disease.SARS virus doesn't induce the maturation of DC.The increase in the size of macrophages indicated that they were in an activated state and may play a role in the pathogenesis of SARS.

lymphocyte,and especially the small lymphocytes were moreheavily labeled than the larger ones,and the Con A receptor of NK cell exhibitedpolarization along the cell surface.These results suggest that:1.the extent of ConA receptors was related to the functional activities of the ceils,the more active thecell is,the more the Con A receptors will be;2.the receptors of Con A wereprobably related to the cell recognition,for instance,on the NK cell Con A rece-ptors showed polar distribution;3.the number of receptors of Con A was dependenton the cell differentiation,the more mature the cell is,the more the Con Areceptors are.

Objective To observe the morphologic changes of immature dendritic cells(imDCs) before and after the electransfection of human hepatic cancer cell RNA. Methods Monocytes were purified from human peripheral blood,and induced into imDCs.Then human hepatic cancer cell RNA was electransfected into monocyte-derived imDCs.ImDCs were identified by the immunocytochemical method with 7 specific antibodies before and after electransfection.These dendritic cells were observed by scanning electron microscopy. Results After electransfection of human hepatic cancer cell RNA there were few changes of molecule expressions in imDCs.ImDCs were in round,oval and irregular shapes before electransfection.Their sizes were not identical but all bigger than monocytes.There were many protrusions in different shapes which looked like dendrite,or/and bubble,veil cloud on the surface of these imDCs.Although there were some cell fusions and cell deaths after electransfection,most imDCs recovered from the damage.Electransfecting human hepatic cancer cell RNA into imDCs would make pores on cell membrane.Conclusions The pores on cell membrane make it possible that the exogenous material enters imDCs.This study can prove the possibility of electransfecting human hepatic cancer cell RNA into imDCs to make cancer vaccine,which provides a new way for tumor biologic therapy.

The effects of the Biological Suppressor of Cancer(BSG)on human normal hone marrow cells and leukaemic cells were studied.There are some evidences that the BSC possesses a biological activities which apparently kill leukaemic cells and inhibit their DNA synthesis,but it has no such effects on erythroblast.The BSC extracted from ascites of ascitic tumors shows no cancer lines specificity,nor tissue specificity.

Objective To investigate the histochemical changes of 3?-hydroxysteroid dyhydrogenase(3?-HSD), sucinic dehydrogenase(SDH) and lipid on rat adrenal cortex in the carcinogenesis-resistance with selentum. Methods Wistar rats stomach carcinogenesis (formation of aneuploid cells in the glandular stomach mucosa) was induced by MNNG(N-methy1-N-nitroso-guanidine,20 mg/kg)gavage. The changes of the adrenal cortex were studied by histochemistry and image analysis during preventing glandular stomach cancer with selenium(0.1 mg/kg and 2.0 mg/kg) Results 1.Dietary Se inhibited the aneuploid cell formation,which induced by MNNG gavage in Wistar rat stomach; 2.The histochemical reactions of 3?-HSD and SDH were both significantly stronger in experimental group than that in normal control group in the course of the rat stomach carcinogenesis(P