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 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 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

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 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.

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

BACKGROUND: Preliminary study has proven that adult human bone marrow-derived mesenchymal stem cells (MSCs) suppress peripheral blood lymphocytes proliferation.But the mechanism was still to be investigated.OBJECTIVE: To study the negatively regulatory effect of adult human MSCs on allogeneic lymphocyte proliferation by cell-free condition.DESIGN,TIME AND SETTING: Cytological observation in vitro,which was performed in the Lanzhou General Hospital,Lanzhou Military Area Command of Chinese PLA between October 2005 and December 2007.MATERIALS: The bone marrow sample was provided by the allo-transplantation donor.The peripheral blood lymphocytes were provided by the healthy volunteer.METHODS: Adult human MSCs were separated with Percoll + adherence method.Allogeneic peripheral blood lymphocytes were obtained from healthy donors with Ficoll solution and the cell concentration was adjusted as 2×109/L for use.100μ L MSCs culture supernatant was taken out in 96-well plates.The groups were following: A superuatant + 3-day MSCs culture media (100 μ L/well); B superuatant + phytohemagglutini (PHA; 1 g/L,5 μ L); C medium + LG-DMEM culture media containing 10% fetal bovine serum (100 μ L); D medium + PHA (1 g/L,5 μ L).The cells were incubated at 37 ℃ with 5% CO2 in a fully humidified atmosphere for three days.MAIN OUTCOME MEASURES: Effect of MSCs supematant on proliferation and transformation of variant lymphocytes.RESULTS: Peripheral blood lymphocytes proliferation was suppressed as compared with the blank control group and PHA group after MSCs culture,and the inhibition ratio was 9.00% (P ＜ 0.05).When lymphocytes were stimulated by PHA,the suppression effects were even stronger and the inhibition ratio was 20.91% (P ＜ 0.01).CONCLUSION: Adult human MSCs supernatant can suppress peripheral blood lymphocytes proliferation and transformation; furthermore,PHA can enhance the inhibitory effect,suggesting the negative regulation is at least in part due to indirectly inhibiting lymphocytes via soluble cytokines.

OBJECTIVE: To explore the curative effect of nasal type extranodal NK/T-cell lymphoma by sequential chemotherapy combined radiotherapy compared with chemtherapy. METHOD: Fifty-seven cases of nasal type extranodal NK/T-cell lymphoma confirmed by pathological morphology and immuno-histochemistry were divided into chemotherapy combined radiotherapy group (34 cases) or chemotherapy group (23 cases). Twenty-three patients in the chemotherapy group alternately applied CHOP, VDLP and MEOP regimen after each two treatments into the clinical observation; Chemotherapy combined radiotherapy group of 34 patients, in addition to the above chemotherapy, applied three-dimensional conformal radiation therapy of the primary site by linear accelerators. Then all of patients were ceased treatment and followed up 3-5 years. RESULT: (1) After treatment, effective rate of two groups was 91.2% and 87.0%, there was no obvious difference (P > 0.05); After follow up about 1 year, effective rate of two groups was 76.5% and 73.9%, there was no obvious difference (P > 0.05); (3) After follow up about 3 years and 5 years, disease free survival (DFS) of two groups was 61.3%, 47.6% and 43.5%, 21.4%, there was obvious difference (P < 0.05). (4) Long-term survival is closely related to treatment mode. (5) B symptoms, advanced (III, IV) stage, the International Prognostic Index (IPI), KPS scores were correlated with prognosis, and were independent prognostic factors. CONCLUSION Treatment with chemotherapy and radiotherapy for nasal type extranodal NK/T-cell lymphoma had obvious curative effect and may improve long-term survival efficiently compared with chemotherapy alone.
Adult ; Antineoplastic Combined Chemotherapy Protocols ; Combined Modality Therapy ; Female ; Humans ; Lymphoma, Extranodal NK-T-Cell ; drug therapy ; radiotherapy ; therapy ; Male ; Middle Aged ; Radiotherapy ; methods ; Treatment Outcome ; Young Adult

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.