Relapse, which puzzled several generations of hematologists, is the bottle-neck of radical treatment for leukemias. The progress of Human Microbiome Project at the beginning of 21st century suggested that human body was a super-organism constituted by the core of human cells and symbiotic microorganisms. The elucidation and characterization of endogenous retrovirus and prion protein suggested the possible effects of co-evolutional microorganisms on human health. Recently, the elucidation of the roles of tunneling nanotubes in intercellular communication and transportation suggested a novel way for cellular communication and transport of oncogenic materials. The role and significance of in vivo cell fusion have been studied in more detail. On the other hand, donor cell leukemia was reported. All of these approaches provide novel insights for studying the mechanism of leukemia relapse. Based on previous work, the authors suggest the hypothesis: there are two possible mechanisms for the relapse of leukemias: the minimal residual disease (MRD) and intercellular transportation of oncogenic materials.
Cell Fusion ; Humans ; Leukemia ; pathology ; Neoplasm, Residual ; pathology ; Recurrence

OBJECTIVETo study melanoma cell fusion and find a highly efficient fusion method for tumor cells.

METHODSMelanoma cells were labeled with green fluorescent protein and red fluorescent protein, respectively, and fused by a modified phytohaemagglutinin (PHA)-ECM830 fusion method. Melanoma fusion cells were selected by the fluorescence activated cell sorting. DNA content was determined by propidium iodide staining.

RESULTSMelanoma cells were labeled with green fluorescent protein and red fluorescent protein markers and successfully fused through PHA-ECM830 fusion method. The fusion efficiency (7.18%) was much higher compared with ECM830 electricfusion method (0.50%). Melanoma fusion cells were successfully obtained by the fluorescence activated cell sorting.DNA content was doubled in melanoma fusion cells compared to B16-F10 melanoma cells. The proliferation rate of melanoma fusion cells was significantly decreased in vitro and in vivo.

CONCLUSIONSWe successfully obtained the melanoma fusion cells by the improved PHA-ECM830 fusion method. The proliferation rate of melanoma fusion cells dramatically decreases.

Animals ; Cell Fusion ; methods ; Cell Line, Tumor ; Cell Proliferation ; Melanoma, Experimental ; pathology ; Mice ; Phytohemagglutinins ; pharmacology

Bioseperation, cell cultivation and cell electrofusion are three main biological processes in space laboratories. Microgravity is free from the influences of convection and sedimentation. Therefore, it is an ideal realm for cell electrofusion and hence it can be used in the research of monoclonal antibody, cross breeding and microgravity biology. This paper reviews the research of cell electrofusion under microgravity, including the changes of cytoskeleton and the mechanism of cell electrofusion.
Animals ; Cell Culture Techniques ; Cell Fusion ; methods ; Electric Stimulation ; Electroporation ; methods ; Mice ; Microelectrodes ; Weightlessness ; Weightlessness Simulation

STUDY DESIGN: A randomized, controlled animal study. PURPOSE: To investigate the effectiveness of fusion and new bone formation induced by demineralized bone matrix (DBM) strips with jelly strengths. OVERVIEW OF LITERATURE: The form of the DBM can make a difference to the outcome. The effect of different jelly strengths on the ability of DBM to form new bone is not known. METHODS: Forty-eight rabbits were randomized into a control group and two experimental groups. In the control group (group 1), 1.4 g of autologous iliac crest bone was placed bilaterally. In the experimental groups, a high jelly strength DBM-hyaluronic acid (HA)-gelatin strip (group 2) and a low jelly strength DBM-HA-gelatin strip (group 3) were used. The fusion was assessed with manual manipulation and radiographs. The volume of the fusion mass was determined from computed tomographic images. RESULTS: The fusion rates as determined by manual palpation were 37.5%, 93.8% and 50.0% in group 1, group 2, and group 3, respectively (p<0.05). By radiography, the fusion rate of High jelly strength DBM strip was statistically significantly greater than that of the other alternatives (p<0.05). The mean bone volume of the fusion mass as determined by computed tomography was 2,142.2+/-318.5 mm3, 3,132.9+/-632.1 mm3, and 2,741.5+/-380.4 mm3 in group 1, group 2, and group 3, respectively (p<0.05). CONCLUSIONS: These results indicate that differences in the structural and mechanical properties of gelatin that are associated with jelly strength influenced cellular responses such as cell viability and bony tissue ingrowth, facilitating greater bone fusion around high jelly strength implants.
Animals ; Bone Matrix* ; Cell Survival ; Gelatin ; Osteogenesis ; Palpation ; Rabbits ; Radiography ; Spinal Fusion

We represent a pathologically proven case of a four-year-old male patient with renal cell carcinoma associated with Xp11.2 translocation/TFE3 gene fusion, which is rare but more frequent in children or young adults. Computed tomography showed about 2.5 cm size ill-defined mass in the right kidney. The mass was hyperechoic on ultrasound. Magnetic resonance imaging demonstrated a mass with capsular enhancement and diffusion restriction. We present a case of Xp11.2 renal cell carcinoma and provide review of the literature.
Carcinoma, Renal Cell ; Child ; Diffusion ; Gene Fusion ; Humans ; Kidney ; Magnetic Resonance Imaging ; Male ; Young Adult

BACKGROUND: Preoperative autologous donation is one of the most widely used methods of autotransfusion. However donation of predetermined units cannot often be achieved because of falling hematocrit with donations going on. We studied on the minimal effective dosage of recombinant human erythropoietin(rhEPO) used in preoperative autologous donation. METHODS: We conducted randomized trial of rhEPO in 45 adults for posterior spinal fusion procedures who had basal hematocrit less than 40 percent. The patients received either nothing(control group) or rhEPO(25 or 50 units/kg) intravenously twice a week for 21 days, during which time up to 3 units of blood was collected. Patients were excluded from donation when their hematocrit values were less than 33 percent. All patient received iron sulfate(256mg orally three times daily). RESULTS: The mean number of units collected per patient(mean+/-SD) was 3+/-0 for the 50-unit group(P<0.05 when compared with control group), 2.84+/-0.36 for the 25-unit group and 2.67+/-0.49 for the control group. The red cell volume donated by the patients who received rhEPO(347 ml, 325 ml) was greater than that donated by the control group(255 ml, P<0.05). The differences between hematocrits of the first and the third preoperative donations were significantly less in 50-unit group(1.50+/-2.05) and 25-unit group(1.51+/-1.85) than that of control group(3.73+/-1.66). Two patients of the 25-unit group and 5 patients of the control group required additional homologous blood postoperatively. And there were no significant differences in the pattern of postoperative changes of hemoglobin and hematocrit among each group. CONCLUSIONS: Fifty units per kilogram of body weight is considered appropriate to be the minimal effective dosage for preoperative autologous donation.
Adult ; Blood Transfusion, Autologous ; Body Weight ; Cell Size ; Erythropoietin* ; Hematocrit ; Humans* ; Iron ; Spinal Fusion

Fractures in ankylosing spondylitis (AS) are often difficult to treat and surgical treatment may be fraught with complications. We describe the use of a robotic-assisted device in the surgical treatment of an unstable L1 fracture in an elderly patient with chronic lymphocytic leukemia and AS. The postoperative course was uneventful and the patient was discharged after 3 days. The use of a robotic-assisted device in spine surgery is particularly indicated in difficult high risk cases.
Aged ; Humans ; Leukemia, Lymphocytic, Chronic, B-Cell ; Spinal Fusion* ; Spine ; Spondylitis, Ankylosing*

BACKGROUND: Immunotherapy using dendritic cells (DC) loaded with tumor antigens may represent a potentially effective method for inducing antitumor immunity. We evaluated the effectiveness of DC-based antitumor immune response in various conditions. METHODS: DC were cultured from peripheral blood mononuclear cells (PBMNC) in myelogenous leukemia (ML) and lysates of autologous leukemic cells are used as tumor antigen. The effectiveness of interleukin-12 (IL-12) and CD40L (CD154) on the antigen presenting function of lysates-loaded DC was analyzed by proliferation, cytokine production, and cytotoxicity tests with activated PBMNC (mainly lymphocytes). For generating antigen-loaded DC, direct fusion of DC with ML was studied. RESULTS: Antigen loaded DC induced significantly effective antitumor immune response against autologous leukemic cells. Administration of IL-12 on the DC based antitumor immune response showed higher proliferation activity, IFN-gamma production, and cytotoxic activity of PBMNC. Also, fused cell has a potent antitumor immune response. CONCLUSION: We conclude that lysates-loaded DC with IL-12 may be effectively utilized as inducer of antitumor immune reaction in ML and in vivo application with DC-based antitumor immunotherapy or tumor vaccination seems to be feasible.
Antigens, Neoplasm ; CD40 Ligand ; Cell Fusion ; Dendritic Cells* ; Humans ; Immunotherapy ; Interleukin-12* ; Leukemia ; Leukemia, Myeloid* ; Vaccination

OBJECTIVEThis study was to establish a stable, effective and reproducible human acute promyelocytic leukemia model in severe combined immunodeficient (SCID) mice by using NB4 cell line, and to investigate the disease course character and biological behaviors.

METHODSThree-five-week-old SCID beige mice were divided randomly into two groups: experimental and control group. SCID mice of experimental group were transplanted by tail vein (iv) injection of 5×10(6) NB4 cells. The WBC cell count and the positive rate of promyelocytes in peripheral blood were dynamically monitored by using smears. Morphological examination and histopathological assay were employed to confirm NB4 cell infiltration in organs (liver, spleen, lung, kidney and brain). The expression level of PML-RARα fusion protein was detected by Western blot.

RESULTSWithin two weeks there was no significant difference in peripheral blood WBC count between two groups (P > 0.05), meanwhile, NB4 cells were not found. At the day 21 and 28 after inoculation, the peripheral blood white blood cell count of experimental group reached to (4.79 ± 1.13)×10(9)/L and (7.62 ± 2.24)×10(9)/L respectively, which were significantly higher than that in control group (P < 0.05); simultaneously, the positive rates of promyelocytes on smears were (2.14 ± 0.63)% and (6.6 ± 2.76)%, respectively. Morphological observation showed single or multiple tumor lumps at day 21 after inoculation; HE staining of tissue biopsies demonstrated a large number of promyelocyte in the liver, spleen, lung, kidney and brain tissue. Cell immunofluorescence results showed that the CD33 expression of bone marrow cells in mice of experimental group was strongly positive (P < 0.05). Western blot confirmed that the PML-RARα fusion protein was expressed variously in liver, kidney and brain tissue.

CONCLUSIONSThe human acute promyelocytic leukemia SCID mouse model is succesfully established by tail vein injection of NB4 cells. This model can mimic the characters of involved bone marrow and diffuse growth of cells. This model is a useful tool to explore the pathogenic mechanism and experimental treatment of human leukemia.

Animals ; Cell Line ; Granulocyte Precursor Cells ; Humans ; Leukemia, Promyelocytic, Acute ; Mice ; Mice, SCID ; Oncogene Proteins, Fusion

OBJECTIVETo investigate the inhibitory effect of RNA interference on chronic myeloid leukemia (CML) bcr/abl oncogene expression.

METHODSThe small interference RNAs (siRNAs) were synthesized in vitro. K562 cells stably expressing bcr/abl gene were transfected with the siRNA by electroporation, both the non-transfected cells and non-specific siRNAs transfected cells were taken as controls. The enhanced green fluorescent protein (EGFP) plasmid was used as positive control and the transfection efficiency was detected by flow cytometry. Inhibitory effect of siRNAs was demonstrated by real-time quantitative RT-PCR and Western blots. Cell proliferation was measured by MTT assay and apoptosis by Annexin V-FITC assay.

RESULTSThe transfection efficiency was about 70%. The synthesized siRNAs inhibited CML bcr/abl oncogene expression at both mRNA and protein levels. siRNAs could inhibit K562 cell proliferation to 47% and 56% at 24 h and 48 h after transfection, respectively, and induce cell apoptosis from 1.00% in control group to 15.05% and 19.4% at 24 h and 48 h respectively.

CONCLUSIONAt the cell level, inhibition of CML bcr/abl oncogene expression by chemically synthesized siRNAs provides the new method for anti-leukemia study.

Apoptosis ; genetics ; Cell Proliferation ; Fusion Proteins, bcr-abl ; genetics ; Humans ; K562 Cells ; RNA, Small Interfering ; Transfection