Objective To approach the dynamic changes of CD~+_ 34 cells and T lymphocyte subsets and best time of peripheral blood stem cell harvest when using rHuG-CSF for peripheral blood stem cell mobilization in donors and patients.Methods From 2001 to 2002 12 donors and 16 patients in Lanzhou General Hospital who received chemotherapy 7 days ago were injected rHuG-CSF 300 ?g/d for mobilization peripheral blood stem cells,and flow cytometry were used to detect the changes of CD~+_ 34 cells and T lymphocyte subsets for 5 days.Results (1)Before using rHuG-CSF,there was obvious difference between patients and donors in bone marrow CD~+_ 34 cells(P

Objective To study the immunomodulatory functions of adult human bone marrow-derived mesenchymal stem cells in vitro.Methods Adult human BMMSCs were separated with Percoll((1.073) g/ml) and cultured in Dulbecco's Modified Eagle's Medium with low glucose containing 10% fetal calf serum.The purity of BMMSCs was identified with the spindle-fibroblastic morphology by microphotograph and the phenotypes were tested by fluorescence-activated cell sorter(FACS).BMMSCs were plated in 96-well plates.After treated with mitomycin,BMMSCs were co-cultured for 72 h with allogeneic lymphocytes isolated from peripheral blood.Effects of MSCs on PHA induced lymphocytes transformation were investigated.The proliferation of lymphocytes was measured by MTT method.Results Peripheral blood lymphocytes proliferation was suppressed by BMMSCs and the inhibition ratio was (60.68%)(P

Objective To investigate the effect of immunocompetent cells and immune regulator on the apoptosis of human mesenchymal stem cells ( MSCs) and on mRNA expression of heme oxygenase-1. Methods MSCs were cultured by density gradient centrifugation and then identified by flow cytometry. RT-PCR was used to detect HO-1 mRNA expression and flow cytometry was used to analyze cell apoptosis after the stimulation of IFN-? and PHA-activated T cells. Results The mRNA expression of Heme oxygenase-1 was observed in MSCs and decreased after the stimulation of IFN-? and activated T cells. IFN-?,znpp-Ⅸ and combined these two caused obvious cell apoptosis in MSCs,with an apoptotic rate of ( 56. 50 ? 0. 16) % ,( 56. 85 ? 2. 27) % ,and ( 82. 53 ? 2. 65) % respectively. All of them had a significant difference compared with the normal MSCs [( 7. 56 ? 1. 43) % ,P

Objective To explore the feasibility and safety of cotransplantation of autologous bone marrow-derived mesenehymal stem cells (MSCs) and peripheral blood stem cells in hematological malignant diseases and to observe its effect on hematopoietic reconstruction after cotransplantation. Methods Adult human MSCs were isolated from the healthy bone marrow of the patient himself with Percoll (1. 073 g/ml) and cultured in Dulbecco's modified Eagle's medium with low glucose containing 10% AB type human serum. After conditioning regimen of high-dose chemotherapy and radiotherapy, cotransplantation of autologous bone marrow-derived MSCs and peripheral blood stem cells was done in five patients with hematological malignant diseases. Results The process of the infusion was safe and there were no adverse reactions or other toxicities related to the infustion of MSCs. The median time to achieve neutrophil counts greater than 0. 5 × 109/L was 9.4 days ( ranging from 8 to 11 days) after cotransplantation and platelet counts greater than 20 × 109/L 12. 2 days (ranging from 10 to 14 days). Conclusion Cotransplantation of autologous bone marrow-derived MSCs and peripheral blood stem cells in hematological malignant diseases is feasible and safe. The rapid hematopoietic reconstruction after cotransplantation shows that MSCs have an effect on hematopeiesis, but the mechanism is still to be investigated.

BACKGROUND:The incidence rate of peripheral T cel lymphoma is high in Asia, and peripheral T cel lymphoma is aggressive with generaly poor prognosis. However, there is no standard treatment strategy. OBJECTIVE:To retrospectively analyze the therapeutic effect of autologous hematopoietic stem cel transplantation on peripheral T cel lymphoma as wel as relevant toxic and side effects. METHODS:A retrospective review was conducted in 35 patients with peripheral T cel lymphoma who underwent autologous hematopoietic stem cel transplantation from March 2003 to April 2014, including 22 cases of extranodal NK/T-cel lymphoma (nasal type), 1 case of angioimmunoblastic T-cel lymphoma, 8 cases of peripheral T cel lymphoma (non-specific), 3 cases of ALK-positive anaplastic large cel lymphoma, and 1 case of ALK-negative anaplastic large cel lymphoma. Al of 35 patients were classified pathologicaly according to WHO pathological type in 2001 and 2008, and received the high-dose chemotherapy with vincristine, cytarabine, etoposide, mitoxantrone, semustine, cyclophosphamide, and total body irradiation. RESULTS AND CONCLUSION: After a median folow-up of 54 (9-120) months, the probabilities of overal survival and disease-free survival after transplantation were 80% (n=28) and 71% (n=25), respectively. Eight cases (23%) relapsed after transplantation, seven of which died. It was safe with mild and moderate transplantation related side-effects on opportunistic infections, oral cavity mucosa and bladder responses and so on, and there were no severe, life-threatening late complications. Autologous hematopoietic stem cel transplantation may be an effective and safe treatment for peripheral T cel lymphoma, and there is a better benefit in peripheral T cel lymphoma patients with first complete remission.

Objective To approach curative effect of using DLI +IL-2 as immunobiotherapy after Mixed-HSCT in acute myelogenous leukemia.MethodAfter times of chemotherapy,8 cases of patients with acute myelogenous leukemia received Mixed-HSCT,then were treated with DLI +IL-2 for 2-7 times.Observed clinical effect for 1 to 5 years.Result DFS in 8 cases of patients with acute myelogenous leukemia received Mixed-HSCT and treated with DLI +IL-2 for 2-7 times were 62.5%.There were no GVHD.Conclusion Immuno-biotherapy with DLI +IL-2 after Mixed-HSCT in patients of acute myelogenous leukemia may be a method to increase DFS efficiently.

Objective To explore the dynamic changes of CD34 + cells and T lymphocyte subsets and best time of harvesting peripheral blood stem cell when G-CSF was used for peripheral blood stem cell mobilization in donors and patients. Methods A total of 12 donors and 16 patients who received chemotherapy for 7 d were injected G-CSF of 300 ?g/d to mobilize peripheral blood stem cells for 5 d and flow cytometry were used to detect the changes of CD34 + cells and T lymphocyte subsets everyday for 5 d. Results ① Before G-CSF treatment, there were obvious differences in bone marrow CD34 + cells between patients and donors (P

Objective To study the immunoregulatory effects of bone marrow mesenchymal stem cells (MSC) on allogeneic peripheral B lymphocytes in vitro. Methods MSCs were isolated and cultured from bone marrow by gradient centrifugation. Mononuclear cells were isolated routinely from peripheral blood, then monocytes were eliminated by L-leucine methy ester method. Remained T lymphocytes were eliminated by AET-SRBC rosette method. The action of MSCs and its supernatant on B lymphocytes proliferation in the presence of anti-human IgM goat antibodies (anti-IgM) was investigated by MTT. The IgG, IgM in the supernatant were detected by ELISA. The percent of apoptosis B lymphocytes, co-cultured with MSCs for 24 or 48h, was assayed by FACS. Results MSCs and its supernatant inhibited B lymphocytes proliferation and Ig secretion. The inhibitory effect depended on the amount of MSCs and condition of its supernatant. The date of FACS indicated that the apoptosis ratio of B lymphocytes, co-cultured with MSCs for different times, were non-significant. The inhibitory effect of MSCs on B lymphocytes was temporary and reversible. Conclusion MSCs have immunoregulatory effects on B lymphocytes, and its mechanisms are complex, not only correlating with the concentration of MSCs but also the action between cells and the secretory cytokine of MSCs.

BACKGROUND:Studies have shown that paeoniflorin functions as replenishing blood and treatment of autoimmune diseases, and bone marrow mesenchymal stem cels also play an important role in the body’s blood and immune function. However, paeoniflorin effects on bone marrow mesenchymal stem cel proliferation and cytokine secretion and expression are rarely reported. OBJECTIVE:To investigate the effect of paeoniflorin on proliferation of bone marrow mesenchymal stem cels and the expression of interleukin-6. METHODS:Human bone marrow mesenchymal stem cels were separated and culturedin vitro by density gradient centrifugation combined with attachment method. The biological characteristics of bone marrow mesenchymal stem cels were identified by flow cytometry and osteogenic/adipogenic induction. The proliferation of bone marrow mesenchymal stem cels under different concentrations of paeoniflorin was detected by MTT method. The mRNA expression and secretion of interleukin-6 in the supernatant of bone marrow mesenchymal stem cels were detected by RT-PCR and ELISA, respectively. RESULTS AND CONCLUSION: Bone marrow mesenchymal stem cels were isolated successfuly and had osteogenic and adipogenic differentiation potential. Compared with the controlgroup, 2 μmol/L and 10 μmol/L paeoniflorin significantly promoted the proliferation of bone marrow mesenchymal stem cels. 10 μmol/L paeoniflorin could significantly decrease the proportion of bone marrow mesenchymal stem cels in G0/G1 phase and increase this proportion in S phase. Compared with the control group, the experimental group could significantly increase the secretion and mRNA expression of interleukin-6 (P < 0.01). It is concluded that paeoniflorin at certain concentrations can obviously promote the proliferation of bone marrow mesenchymal stem cels, and increase the expression and secretion of interleukin-6.

Objective To investigate the curative effect of allogeneic hematopoietic stem cell transplantation (allo-HSCT) and the method of the pretreatment choice on myelodysplastic syndrome (MDS).Methods 13 cases who were undergoing allo-HSCT including 10 HLA-matched, 2 HLA partially matched,and one umbilical cord blood transplanted patients were enrolled in this study. The infusion number of MNC and CD+34 cells is 6.92 (2.65-21.33)×108/kg and 4.47 (1.49-10.22) ×106/kg, respectively. Of the 13 cases,5 adopted TBI, fludarabine(Flud), and cyclophosphamide(Cy) pretreatment, 3 chose BU/Cy pretreatment, and 3 chose TBI and Cy pretreatment, 2 chose Ara-C+BU+Cy+VM26 pretreatment. In order to prevent GVHD, 2cases of HLA partially matched were treated with ATG, cyclosporin A (CsA), MTX and MMF aftertransplantation, while the others were treated with CsA and MTX only. Results 9 out of 13 cases achieved hematopoietic reconstruction completely. 4 died of complications. Conclusion The allogeneic hematopoietic stem cell transplantation is an efficient regimen to cure MDS. The pretreatment should adopt the individuation.