The Inducement Of Qing Du Fu Zheng Recipe On DC From BMNC Of ANLL-CR Patients

PartⅠLiteratureAcute leukemia (AL) is one of the common hematology tumors, which has some characteristics, as like with serious condition and the fast development, easy to relapse after remission, low long-term survival rate. As the improvement in Chinese medicine and Western medicine theory, continuous development of the treatment to leukemia is keeping up so that the rate of acute leukemia in complete remission (CR) and long-term survival is with a considerable degree of improvement or even cure. However, most patients still relapse after complete remission, for which the fundamental reason is the existence of minimal residual leukemia cells. To control such a state of minimal residual leukemia (MRD) is one of the hotspots of leukemia research.The international consensus is going like this; the matter of immune function besides, the presence of minimal residual leukemia has more to do with leukemia cell immune escape mechanisms, so that to kill the minimal residual leukemic cells must rely on how the body's own immune surveillance functions are performing. In this stage, after surviving a long-term chemotherapy, drug sensitivity of leukemic cells has become very low and this is the best time for the body to kill residual tumor cells assisting by immune therapy aids.Based on the knowledge about pathogenesis, such like righteousness weak, cult poison invasion of, Chinese Medicine for minimal residual disease at present has the basically same understanding of the pathogenesis. The body is on "evil to being bad," "righteousness weak" and "not entirely over evil "pathophysiological state. PartⅡExperimental study Qingdu Fuzheng recipe on ANLL-CR patients BMNC induction of DC source1. Research purposesIn this experiment, Qing du Drink and Yang zheng Troche in acute myeloid leukemia patients in remission bone marrow mononuclear cells (BMNC) were cultured in vitro to observe the dendritic cells (DC) induced transformation of the situation. To induce transformation of normal DC and purified T cells were cultured with autologous T cells after stimulation was observed after stimulating T cells against human leukemia cell line K562 cells in vitro.Qing du Drink and Yang zheng Troche are the effective hospital treatment of acute leukemia which is developed by our hospital. This study is researching from the new ways of treatment of leukemia combined by Chinese medicine, and from a new entry point that is to increase remission in patients with antigen presenting ability of dendritic cells. The purpose is to look for the effective ways for leukemia remission but to prevent its recurrence.2. Method2.1 The source of specimen2.1.1 Bone marrow mononuclear cells derived from the Guangzhou University of TraditionalChinese Medicine Department of Hematology, First Clinical Medical College of patients hospitalized with acute myeloid leukemia after complete remission (AML-CR) of bone marrow in two cases. Patients meet diagnostic criteria for acute leukemia after complete remission after standard chemotherapy, consistent with acute leukemia in complete remission criteria.2.1.2 T cells from normal adult peripheral blood and peripheral blood of leukemia patients with complete remission after one each.2.1.3 Leukemia cell line K562 cells.2.2 Preparation of herb medicineGroup of New Zealand rabbits, respectively Qingdu drink, keep positive, keep positive Qingdu drink and by low, medium and high doses of fed animals, preparation of the three groups with different concentrations of serum. 2.3 Separation of bone marrow mononuclear cells to T cellsApplication procedures for sheep erythrocyte rosette separated from the bone marrow T cells in the bone marrow mononuclear cells (TD-MNC).2.4 Training from AML-CR patients with TD-MNC in DCThe TD-MNC cells with different Chinese medicines containing serum and cytokines in joint training, divided into:drinking group Qingdu, GM-CSF IL-4+IFN-α+TNF-αgroup and disinfection drink+GM-CSF+IL-4+IFN-α+TNF-αgroup, support positive group, the raising of positives+GM-CSF+ IL-4+IFN-α+TNF-αgroup and disinfection drink and keep positive+GM-CSF +IL-4+IFN-α+TNF-αgroup. Equalization of the group with serum is low and medium and high dose group.2.5 DC identification includemorphology, immunological detection, and the secretion of cytokines IL-2, IL-6 determination.2.6 Purification of peripheral blood T cells (magnetic activated cell sorting method)Normal, AML-CR patients are depicting a blood 50ml, by Ficoll density gradient centrifugation of peripheral blood mononuclear cells (PBMNC), the use of bead negative selection method were purified peripheral blood T cells.2.7 allogeneic T cells and stimulate autologous T cellsWere collected purified allogeneic T cells, AML-CR T cells from patients with AML-CR patients with TD-MNC derived DC to 10:1 ratio by adding 24-hole incubated 5d, medium supplemented with 100U/ml of IL-2.2.8 stimulate T cells to K562 Cells (MTT)T cells in each group with K562 cells, respectively 10:1,20:1,40:1 effect: target ratio of culture based on cell kill rate.3. The resultsThe experimental results show that the group of Qing du Drink and Yang zheng Troche combined with cytokines can promote DC expression of CDla, HLA-DR, CD80, CD86, and to promote DC high secretion of cytokines IL-2, IL-6 is significantly higher than the cell group, which is statistically significant. Also, the Chinese medicine group is higher than the cell group on efficiency butt and the killing rate. The Chinese medicine group also explains the greater the concentration of T cells, tumor cell-killing rate is increased. The Chinese group compared with the control group shows that the concentration of medicine in a joint cell factor can be better than simply promote the use of DC cytokines.PartⅢClinical ObservationAttachment:Qing du fu zheng recipe on AL-CR in patients with bone marrow cells of CD antigen expression and proliferation1. Research purposesWe observe the expression rate of CD34, CD34/CD2, CD34/CD19, CD13/CD33 of acute leukemia patients' bone marrow cells after complete remission to understand the effect of the treatment for tiny residual leukemia; To compare the difference of bone marrow cell proliferation cycle between before and after treatment, we try to deeply study the possible mechanism of this treatment to tiny residual leukemia.2. Research MethodsThis study adopts random comparison to observe Qing du Drink and Yang zheng Troche's impact on bone marrow cells CD antigen's expression and proliferation cycle of acute leukemia patients after the complete remission.2.1 Case from Traditional Chinese Medicine from the University Hospital and Department of Hematology, First Affiliated Hospital outpatients.2.2 treatments2.2.1 control:according to conventional chemotherapy to consolidate maintenance treatment (ANLL with the DA or MA or HA program, ALL or VDLP program with VDP) in chemotherapy, such as infection, bleeding, etc. according to conventional anti-infection and hemostasis.2.2.2 Observer Group:Traditional Chinese principle of treatment with the control group. Consolidation chemotherapy, while maintaining oral Qing du Drink and Yang zheng Troche, Japan 1, sub-second service, for 4 weeks. 2.2.3 The two groups were collected before treatment and four weeks after the bone marrow, experimental observation.2.3 Detection of bone marrow cells the expression of CD antigens by flow cytometry before and after treatment in patients with bone marrow cells CD34, CD34/CD2, CD34/CD19, CD13/CD33 expression.2.4 Detection of bone marrow cell cycle by flow cytometry before and after treatment of bone marrow cell proliferation cycle in the expression.3. Experimental resultsThe study combined with the measurement of patient's bone marrow cell cycle by flow the cytometry machine indicates that Qing du Drink and Yang zheng Troche have the capability of restraining the CD34's expression, indicates that Qing du Drink and Yang zheng Troche might restrain the proliferation of leukemia cells so that it could decrease the expression rate of CD13/CD14/CD33, which could represent myeloid leukemia cells and CD3/ CD19, which could represent lymphobiastic leukemia cell.PartⅣConclusionsThe research on Chinese monomer or compound in leukemia tumor antigen presenting has not been seen yet. This paper firstly used the serum of pharmacology to study compound prescription drug drinking clean, keep positive synergistic cytokines to Inducement of bone marrow mononuclear sourcing dendritic cells of acute leukemia and its biological characteristics after complete remission and observe the effects of clear Fuzheng drug compound in clinical to bone marrow cells immunophenotype and generation cycle of acute leukemia patients after complete remission, try to discover the mechanism of this treatment to acute myelocytic leukemia after complete remission, in order to make a further improvement of Chinese medicine treatment to this disease.