1.Effect and mechanism of glycosylphosphatidilinoditol- specific phospholipase D on the adhesion function of bone marrow mononuclear cells separated from myeloid leukemia patients
Guangfen XIAO ; Fangping CHEN ; Bin FU ; Guangping WANG ; Zaifu JIAN
Chinese Journal of Tissue Engineering Research 2010;14(1):187-190
BACKGROUND: There still is rarely report about the effect of glycosylphosphatidilinoditol-specific phospholipase D (GPI-PLD) on the adhesion function of leukemic cells through screening Medline and CNKI databases.OBJECTIVE: To observe the effect of GPI-PLD on the adhesion function of bone marrow mononuclear cells separated from myeloid leukemia patients, and to investigate the related mechanism. DESIGN, TIME AND SETTING: This study addressing cytology in vitro was conducted at the Hematological Laboratory of Xiangya Hospital from January to June 2004.MATERIALS: Bone marrow was collected from myeloid leukemia patients at the Department of Hematology, Xiangya Hospital, China.METHODS: The GPI-PLD activity of bone marrow mononuclear cells separated from myeloid leukemia patients was measured by using GPI-anchored placental alkaline phosphatase as substrate and Triton-X114 partition. By use of 1,10-phenanthroline, the activity of GPI-PLD was inhibited, the experiment was divided into 2 groups: treatment group adding phenanthroline to achieve a final concentration of 1 mmol/L, while control group adding the same amount of phosphate buffered saline. The adhesion rate to the fibronectin and CD24 expression of these cells were measured by MTT and immunohistochemical method, respectively.MAIN OUTCOME MEASURES: GPI-PLD activity of myeloid leukemic cells, cell adhesion rate, CD24 expression were all measured. RESULTS: The GPI-PLD activity of bone marrow mononuclear cells separated from myeloid leukemia patients was inhibited significantly after these cells were treated by 1 mmol/L 1,10-phenanthroline for 5 hours compared with control groups [(5.40±2.96)%, (42.08±7.21)%, P < 0.01]. At the same time, the adhesion rate of these cells were increased after the GPI-PLD activity was inhibited [(61.19±29.14)%, (49.78±26.73)%, P < 0.01], and the CD24 expression was also up-regulated [(18.5±11.14)%, (16.02±9.68), P < 0.01].CONCLUSION: The adhesion rate of bone marrow mononuclear cells separated from myeloid leukemia patients can be promoted by inhibiting GPI-PLD activity. At the same time, the CD24 expression of GPI-anchored proteins on bone marrow mononuclear cells is improved.
2.Effect of Teicoplanin to Cure Neutropenic Patients with Infection
Jing LIU ; Xin LI ; Tiebin JIANG ; Guangfen XIAO ; Erhua WANG
Chinese Journal of Nosocomiology 2009;0(22):-
OBJECTIVE To investigate the effect and safety of teicoplanin to cure the neutropenic patients with infection.METHODS The 64 neutropenic patients with fever were treated with teicoplanin for empirical therapy,and then its effect and safety were evaluated.RESULTS Teicoplanin was effective in 53 patients,its effective rate was 88.3%,the adverse event rate was only 4.7%,and the adverse reactions were slight and reversible and it was well tolerant to patients.CONCLUSIONS Teicoplanin is one of the antibiotic agents with high performance,low toxicity and fast effect,which is an ideal agent for empirical therapy for neutropenic patients with fever in hematology units.
3.Glycosylphosphatidilinoditol-specific phospholipase D expression in bone marrow mononuclear cells derived from acute leukemia patients
Guangfen XIAO ; Fangping CHEN ; Guangping WANG ; Bin FU ; Junming XIE ; Yingni CHENG ; Qun LI ; Zaifu JIAN
Chinese Journal of Tissue Engineering Research 2010;14(6):1134-1137
BACKGROUND: The correlation of gycosylphosphatidilinoditol-specific phospholipase D (GPI-PLD) activity, mRNA expression to leukemia type, hepatosplenomegaly and/or lymphadenopathy has been rarely reported. OBJECTIVE: To explore the correlation of GPI-PLD expression to leukemia type and hepatosplenomegaly and/or lymphadenopathy of acute myeloid leukemia (AML) patients. METHODS: Fresh bone marrow specimens were obtained from 43 newly diagnosed AML patients, 28 acute lymphocytic leukemia (ALL) patients, and 21 normal persons. Bone marrow mononuclear cells were harvested by density gradient centrifugation. GPI-anchored human placent alkaline phosphatase was used as substrate. GPI-PLD activity was determined bytriton-X114 phase partitioning procedure. GPI-PLD mRNA expression was detected by semi-quantitative RT-PCR. The relationship of GPI-PLD activity, mRNA expression and leukemia type, hepatosplenomegaly and/or lymphadenopathy was analyzed. RESULTS AND CONCLUSION: Compared with control group, GPI-PLD activity and mRNA expression in bone marrow mononuclear cells were significantly higher in AML group (P < 0.01), while they were significantly lower in the ALL group (P < 0.01). Of 43 patients with AML patients, 13 patients had hepatosplenomegaly and/or lymphadenopathy. The GPI-PLD activity (%) and mRNA expression were significantly higher in AML patients without hepatosplenomegaly and lymphadenopathy than those patients with hepatosplenomegaly and/or lymphadenopathy (P < 0.05). These results demonstrated that GPI-PLD activity alteration is consistent with GPI-PLD mRNA expression in AML patients, and the expression levels correlate to leukemia type and hepatosplenomegaly and/or lymphadenopathy of AML patients.
4.Expression of DLK1 gene in acute leukemias and its function in erythroid differentiation of K562 cell line.
Xueyuan TANG ; Chan LONG ; Chenghong WANG ; Guangfen XIAO
Journal of Central South University(Medical Sciences) 2009;34(9):886-891
OBJECTIVE:
To determine the expression of DLK1 gene in acute leukemias (AL) and its function in erythroid differentiation of K562 cells.
METHODS:
We detected the expression of DLK1 gene in 65 different acute leukemia categories (a test group) and 34 normal bone marrow controls (a control group) with RT-PCR. DLK1 protein in 20 out of the 65 AL patients and 13 of the 34 controls was assayed by Western blot. The K562 cell line was induced to erythroid differentiation by hemin. We observed the relationship between its expression and erythroid differentiation.
RESULTS:
Both leukemia cells and normal marrow cells expressed DLK1. The expression of DLK1 mRNA in patients in the test group was higher than that in the control group (P=0.018), while there was no significance between acute lymphoblastic leukemia and acute myelogenous leukemia (P>0.05).The expression of DLK1 mRNA in the test group at onset had no relation with the WBC and platelet count in the total peripheral blood, and the same was true for blast cell rates in bone marrow cells.The level of DLK1 protein in the test group was higher than that in the control group, which was consistent with the mRNA expression (P=0.042). The expression of DLK1 mRNA decreased gradually with K562 cells towards hemin-induced erythroid differentiation.
CONCLUSION
DLK1 gene may be involved in leukemia,but the mRNA level of DLK1 has no relation with some clinical characteristics of AL patients at onset. DLK1 may inhibit the erythroid differentiation of K562 cells.
Acute Disease
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Adolescent
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Adult
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Aged
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Calcium-Binding Proteins
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Case-Control Studies
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Cell Differentiation
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drug effects
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genetics
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Cell Transformation, Neoplastic
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genetics
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Child
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Child, Preschool
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Erythroid Cells
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pathology
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Erythroid Precursor Cells
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pathology
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Female
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Humans
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Intercellular Signaling Peptides and Proteins
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genetics
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metabolism
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K562 Cells
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Leukemia
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genetics
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metabolism
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Male
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Membrane Proteins
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genetics
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metabolism
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Middle Aged
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RNA, Messenger
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genetics
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metabolism
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Young Adult