Study On Immune Function Of DCs And TCs In Peripheral Blood Of Patients With Different Attributes Of Yin And Yang In Patients With Hepatic Failure

Part-ⅠHuman peripheral blood dendritic cells were cultured and identified Purpose:Peripheral blood dendritic cells in vitro method.Method:Under sterile conditions remove the healthy man fresh peripheral blood20ml.Extraction (density gradient centrifugation) mononuclear cells (PBMCs), Take adherent cells in complete medium (containing recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF,)1000U/ml、Recombinant human interleukin4(rhIL-4),500U/ml of recombinant human tumor necrosis factor a (of rhTNF-α)20ng/ml induced culture of DCs, Under the inverted microscope to observe the morphological characteristics of in vitro culture. transmission electron microscopy analysis of the internal structure of the DCs cells, using flow cytometry phenotype molecule expression, and identification.Result:This experimental system can be induced by DCs from peripheral blood PBMCs formation about every20ml (1.3～3.6×106), TEM confirmed that was typical of DCs cell morphology, flow cytometry was used to detect the phenotype of DCs molecular, ELISA kit DCs supernatant of IFN-α, IL-4secretion levels. Expression of DCs, respectively FITC-CD1α44.57±3.12%. PE-HLA-DR90.45±4.37%、 FITC-CD8078.92±2.18%、PE-CD8677.02±1.93%, FITC-CD8340.29±4.17%;Of IFN-alpha, IL-4secretion level of28.56±8.31,37.7710.28.Conclusion:RhGM-CSF and rhIL-4, of rhTNF-α was amplified by a large number of DCs from peripheral blood, and laid the foundation fo research and clinical application of DCs. Part-ⅡExtraction of human peripheral blood T lymphocytesPurpose:The establishment of T lymphocytes in vitro separation and extraction methodMethod:Fresh healthy human blood mononuclear cells separated by density gradient centrifugation after moved to37℃,5%CO2incubator to incubate overnight. Supernatant by adding the T cells in nylon wool column, incubator tending1h, PBS rinse to get the T lymphocytes.Result:Lymphocytes collected directly with PE-labeled mouse anti-human CD3and CD8, FITC-labeled CD4marker, detected by flow cytometry. The results showed normal levels of CD3+, CD4+and CD8+ratio of CD4/CD8expression levels, respectively are70.92±1.77%.42.95±2.01%,24.24±1.35%.1.78±0.07%.Conclusion:In this study, the morphology of T lymphocytes, the performance of preliminary observation and analysis, and to lay the foundation for further in-depth study and clinical application of T lymphocytes. Part-Ⅲ HBV-related liver failure, jaundice, Yin Huang, Yang yellow card and peripheral blood immune cell functionPurpose:In this study, by observing the HBV-related liver failure in patients with jaundice Yin Jaundice, sun yellow card peripheral blood dendritic cells and T lymphocyte function, to understand the HBV-related liver failure, jaundice Yin Jaundice, sun yellow card two disease typeimmune expression of the characteristics.Method:The study included patients with HBV-related liver failure, jaundice, divided into the two groups of sun yellow group Yin Huang group, another a group of healthy controls, PBMCs in peripheral sources in vitro cultured DCs and T lymphocytes induced by application flowtype cell count detection of DCs surface molecules, HLA-DR, CD80, and CD86, CD83, Peripheral circulating CD1α, as well as T-cell surface molecules CD3and CD4and CD8, and detected in the supernatant of DCs with IFN-alpha, IL-4secretion levelsobserve and compare HBV-related liver failure in patients with yin yang yellow card with immune cells and inflammatory factor expression differences.Result1. Of HBV-related liver failure Yang Huang, Yin Huang two syndromes in patients with DCs surface molecules of HLA-DR, Peripheral circulating CD1α, CD83, CD80, and CD86, expression rate, and T cells of CD3, CD4and CD4/CD8expression ratesstatistically significant (P<0.05) compared with healthy people, CD8expression rates and healthy people was significantly different (P<0.01).2. Factor of the HBV-related liver failure in two certificates of patients with type DCs secrete IFN-α alpha compared with healthy people with significant difference (P<0.01), Yin Huang IL-4compared with healthy people statistical difference (P<0.01)3. DCs surface molecules between the2HBV-related liver failure patients yang yellow group, Yin Huang group of HLA-DR, Peripheral circulating CD1α, CD80, no significant (P>0.05), Yin Huang group of CD83, CD86expression rate of less than positivethe yellow group (P<0.01), TCs surface molecules CD3and CD4was no significant difference (P>0.05), yin Huang group, CD8, CD4/CD8ratio of expression of a significant difference (P<0.01) than the sun yellow group.Conclusion:1. DCs of HBV-related liver failure patients, the TCs of the functional state of cellular immune function and inflammation in IFN-α increased performance.2. Yang Huang syndrome DCs, TCs functional status of the performance of cellular immune function and increased inflammation IFN-α3. Yin Jaundice in patients with DCs, the functional state of TCs performance for cellular immune function, inflammation of IFN-α, anti-inflammatory factor IL-4increased4. Yang yellow card, Yin Huang syndrome DCs and the functional status of TCs, indicating Yin Jaundice in patients with cellular immune function is low, Yin Huang syndrome at the same time the existence of inflammatory cytokines IFN-α and anti-inflammatory factor IL-4increased...