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Objective:To investigate the effects of hemophagocytic syndrome also known as hemophagocytic lymphohistiocytosis (HLH) on the clinical features and therapeutic efficacy of patients with Epstein-Barr virus-positive T-cell lymphoma (EBV-TCL).Methods:The clinical data of patients with EBV-TCL diagnosed by pathological examination in the First Affiliated Hospital of Guangzhou Medical University from November 2015 to August 2020 were retrospectively analyzed. According to whether they were accompanied with HLH at the time of onset, patients were divided into HLH group (10 cases) and non-HLH group (13 cases), and the clinical features and prognosis of the two groups were compared. The curative effects of different treatment methods and patients with different plasma EBV-DNA titers were compared.Results:Among 23 patients, 3 cases (13.0%) were in Ann Arbor stage Ⅰ-Ⅱ, 20 cases (87.0%) were in stage Ⅲ-Ⅳ; the International Prognostic Index (IPI) score was 1 point in 3 cases (13.0%), 2 points in 4 cases (17.4%), 3 points in 8 cases (34.8%), 4 points in 8 cases (34.8%). In the HLH group, there were 2 cases of aggressive NK-cell leukemia and 3 cases of childhood systemic EBV-TCL. There were no cases of above two pathological types in the non-HLH group. In the HLH group, the proportions of patients with fever, bone marrow invasion, IPI score > 2 points, and EBV-DNA > 10 4 copies/ml were higher than those in the non-HLH group (all P < 0.05). The objective response rate (complete remission plus partial remission) of all patients after chemotherapy was 47.8% (11/23); there were 3 cases undergoing hematopoietic stem cell transplantation in both the HLH group and the non-HLH group, and all achieved objective remission. The objective remission of 7 patients and 10 patients who did not undergo hematopoietic stem cell transplantation in the HLH group and non-HLH group after lymphoma chemotherapy had 0 case and 5 cases, respectively, and the difference was statistically significant ( P = 0.044). In the chemotherapy alone group, 5 of 17 patients had objective remission, 6 patients in the chemotherapy plus transplantation group had objective remission, and the difference was statistically significant ( P = 0.039). Among 16 patients whose plasma EBV-DNA titers turned negative, 11 patients had objective remission, and 7 patients whose plasma EBV-DNA titers were continuously positive had no objective remission, and the difference was statistically significant ( P = 0.001). The 1-year overall survival rate of all patients was 69.3%, and the 2-year overall survival rate was 52.0%. In the HLH group, the 1-year and 2-year overall survival rates of 7 patients receiving chemotherapy alone and 3 patients receiving chemotherapy plus transplantation were 42.9% and 66.7%, respectively. In the non-HLH group, the 1-year overall survival rates of 10 patients receiving chemotherapy alone and 3 patients receiving chemotherapy plus transplantation were 80.0% and 100.0%, respectively; the 2-year overall survival rates were 26.7% and 100.0%,respectively. The overall survival of patients receiving chemotherapy plus transplantation was better than that of those receiving chemotherapy alone in both the HLH group and the non-HLH group, and differences were statistically significant (all P < 0.05). Conclusions:The general clinical stage of patients with EBV-TCL is later, and the prognosis of EBV-TCL patients with HLH is worse. The therapeutic efficacy may be related to plasma EBV-DNA titers. Hematopoietic stem cell transplantation can improve the remission rate.
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BACKGROUND:Hematopoietic reconstruction of malignant tumor in hematopoietic system is related to disease itself,pretreatment program and therapeutic tool after transplantation;especially,mobilization.collection and cryopreservation of auto-peripheral blood stem cell play a key role in successful reconstruction of hematopoietic system after transplantation.OBJECTIVE:To investigate the reconstruction of hematopoietic system through mobilization, collection and cryopreservation of auto-peripheral blood stem cell in patients with malignant tumor and analyze the effective factors on quantity and quality of auto-peripheral blood stem cell.DESIGN:Case analysis based on malignant tumor in hematopoietic system.SETTING:Department of Blood,Guangzhou General Hospital of Guangzhou Military Area Command of Chinese PLA;Department of Blood,Zhujiang Hospital,Nanfang Medical University.PARTICIPANTS:A total of 18 patients with malignant tumor in hematopoietic system were selected from Department of Blood,Guangzhou General Hospital of Guangzhou Military Area Command of Chinese PLA.Their ages ranged from 1 6to 56 years.Among them,2 patients had acute myelogenous leukemia(AML),1 acute lymphoblastic Ieukemia(ALL),2 lymphoblastic Ieukemia (LL),2 chronic granulocytic leukemia(CGL),4 multiple myeloma(MM),and 7 non.Hodgkin lymphoma.Granulocyte colony-stimulating factor(G-CSF)was made by Chugai Pharmaceutical Company Limited (batch number:N3G31).METHODS:①All patients were mobilized with associated chemotherapy+G-CSF.Associate chemotherapy:Patients with leukemia were given 2 g/m2 arabinosyl cytosine every 12 hours from lhe first to the third days and 200 mg/m2 etoposide or 50 mg/m2 fludarabine from the first to the fifth days. In addition patients with MM were treated with arabinosyl cytosine as the same way mentioned above and with 1 g/m2 cyclophosphamide from the first to the second days. And patients with lymphoma were given 2 g/m2 cyclophosphamide from the first to the second days. When numbers of leucocyte of all patients decreased below 1.0×109L-1 after chemotherapy.G-CSF started mobilization and the collection was stopped with 5μg/(kg·d)subcutaneous injection.②When numbers of leucocyte increased to (4.0-10.0)×109 L-1,hemopoietic stem cells of peripheral blood were collected till the amount of mononuclear cells≥4.0×108/kq or numbers of CD34+ cells≥2.0×108/kg.And then,the samples were dealt with cooling device.maintained in liquid nitrogen at-196℃ and defrosted in water bath at 37-40℃.③Focal sites of patients were pretreated with local irradiation with 200 cGy/time and 5 times/week for 4 successive weeks.The total dosage was 40 Gy.At 48 hours later,(55.3±28.7)mL hemopoietic stem cells of peripheral blood were transfused back. And the duration from transfusion to collection was about(56.5±22.3)days.300 μg/d G-CSF was subcutaneously injected into all patients at 1 day after transplantation and the reaction was stopped at the phase of neutrophil≥0.5×109L-1. Finally. Refusing-staining rate of trypan blue of peripheral blood stem cell, amount of mononuclear cells, number of granulation-monophyly progenitor cell colony and percentage of CD34+ cells were detected before and after thaw.MAIN OUTCOME MEASURES:①Collection of auto-peripheral blood stem cell;②survival rate and related markers of auto-peripheral blood stem cell after cryopreservation;③hematopoietic reconstruction of auto-peripheral blood stem cell after transplantation.RESULTS:All 18 patients with malignant tumor in hematopoietic system were involved in the final analysis.The mean collection time of auto-peripheral blood stern cell was 12.6 days after chemotherapy.the collection times were 1.9.total number of leucocyte was(8.93±1.27)×1 0.L-1 on the first day,and collection rate of mononuclear cell was (138.33±28.61)%. ②Refusing-staining rate of trypan blue of auto-peripheral blood stem cell was similar before and after cryopreservation[(96.26±1.33)%, (92.75±2.04)%,P＞0.05].in addition,after cryopreservation,recovery rates of mononuclear cells,CD34+ cells and granulation-monophyly progenitor cell were(91.96±1.37)%, (85.94±0.64)%and (87.69±4.53)%,respectively.Collection rate of mononuclear cells,number of granulation-monophyly progenitor cell colony and percentage of CD34+ cells were lower in patients with myeloma than in those with leukemia and lymphoma (t=2.524-3.268.P＜0.05).③At 15 days after transplantation,15 patients had the neutrophil≥0.5× 109L-1;at 20 days after transplantation,blood platelet was≥20 × 100 L-1.granulation-monophyly progenitor cells[(18.67-26.82)× 105/kg] of 5 patients grew poorly if the course of chemotherapy was more than 10 times.Among them,3 patients had delayed hematopoietic reconstruction after transplantation of auto-peripheral blood stem cell.CONCLUSION:①High-dose chemotherapy combined with G-CSF can shorten collection time of peripheral blood stem cell and improve collection rate of mononuclear cells.②Increase of chemotherapy times before transplantation can affect quantity and quality of auto-peripheral blood stem cell and cause delayed hematopoietic reconstruction.
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AIM: To study the cytoskeleton of mesench ymal stem cells (MSCs), the ultrastructure and function relationship by using atomic force microscope (AFM). METHODS: The ultrastructures and morphological feature of MSCs c ultured for 1 d and 5 d were studied by AFM. RESULTS: The special structures that possess peculiar morphologi cal characteristic of MSCs such as cytoskeleton, pseudopod, microfilament etc we re identified by AFM, and these special structures are difficult to observe unde r electronic microscopy or other conventional optical microscopy. CONCLUSIONS: AFM is a powerful tool to study ultrastructures, mo rphological features, and cytoskeleton of stem cells in near physiological condi t ions. Its application prospect in cellular biology is extensive. The special cyt oskeleton and other structures of MSCs observed above may represent the structur al base of multi-differentiation potential of MSCs.