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<p><b>OBJECTIVE</b>To explore the correlation between expression of surviving gene in acute leukemic cells and its clinical effects.</p><p><b>METHODS</b>By using semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR) technique, surviving gene expression in 50 previously untreated acute leukemia (AL) patients was analysed. The apoptosis of primary leukemia cells cultured in vitro was assayed with terminal deoxyribonucleotidyl transferase mediated dUTP-biotin nick end labeling (TUNEL).</p><p><b>RESULTS</b>Surviving gene expression levels in cells of AL patients at diagnosis were significantly higher than that in normal bone marrow mononuclear cells (MNCs) (82.0% vs 33.3%, P < 0.05). The expression level was higher in ALL cells than in ANLL cells (89.5% vs 75.0%). Among 22 cases of ANLL, bone marrow remission (BMR) rate was higher in surviving gene negative expression cells from patients accepted a course of chemotherapy than in positive expression cells (83.3% vs 25.0%, P = 0.023). Among 13 ANLL patients received a course of HA regimen chemotherapy, the BMR was higher in patients surviving mRNA negative expression cells than in positive cells (100.0% vs 27.3%). Patients with surviving/beta-actin ratio>0.6 attained lower BMR.</p><p><b>CONCLUSION</b>Higher expression level of surviving mRNA in AL cells may be one of the reasons that leukemic cells are insensitive to chemotherapy.</p>
Subject(s)
Adolescent , Adult , Aged , Female , Humans , Male , Middle Aged , Antineoplastic Agents , Therapeutic Uses , Chromosomal Proteins, Non-Histone , Genetics , Metabolism , Drug Resistance, Neoplasm , Genetics , Physiology , Gene Expression Regulation, Leukemic , Inhibitor of Apoptosis Proteins , Leukemia, Myeloid, Acute , Drug Therapy , Genetics , Metabolism , Microtubule-Associated Proteins , Neoplasm Proteins , Precursor Cell Lymphoblastic Leukemia-Lymphoma , Genetics , Metabolism , RNA, Messenger , Remission Induction , Treatment OutcomeABSTRACT
<p><b>OBJECTIVE</b>To evaluate the effect of mycophenolate mofetile (MMF) on acute graft versus host disease (aGVHD) prophylaxis after allogeneic bone marrow transplantation (allo-BMT) in a murine model.</p><p><b>METHODS</b>Acute GVHD was induced in male BALB/c mice by total-body irradiation (TBI) followed by female allogeneic (C57BL/6J) bone marrow and spleen cells transplantation. Acute GVHD was assessed both physically and histologically. Severe aGVHD was developed in the recipients and the mean survival time (MST) of untreated BM recipients was 6 days. After allo-BMT, animals were divided into 6 groups. Group 1 was given MMF (30 mg.kg(-1).d(-1)), group 2 CsA (1.5 mg.kg(-1).d(-1)) and MTX (0.4 mg.kg(-1).d(-1)), group 3 MMF (30 mg.kg(-1).d(-1)) in combination with CsA and MTX, group 4 MMF (60 mg.kg(-1).d(-1)) in combination with CsA and MTX, group 5 MMF (10 mg.kg(-1).d(-1)) in combination with CsA and MTX, and group 6 no agents for aGVHD prophylaxis. The physical signs, MST, peripheral blood counts, and aGVHD histopathologic examination were observed in all recipients.</p><p><b>RESULTS</b>Animals in control group developed typical aGVHD and 100% of mortality, with a MST of 6 days. The MSTs of group 1 approximately 5 were significantly longer than that of control, being 3.4, 8.4, 9.0, 6.1 and 8.8 days longer, respectively (P < 0.05). The MSTs of groups 2 approximately 5 were longer than that of group 1 (P < 0.05), but there was no statistical significance between groups 3 approximately 5 and 2. There was no statistical difference in peripheral blood count among groups 1 approximately 5. Histopathological examination of skin, liver, and gastrointestinal tract showed typical signs of aGVHD. Animals received immunosuppressive agents (MMF, CsA, MTX) showed the less severe signs.</p><p><b>CONCLUSIONS</b>MMF markedly prolonged MST of allo-BMT recipients, delayed the onset of aGVHD signs. The prophylaxis effect of CsA + MTX with or without MMF was better than that of MMF alone, synergism between MMF of 10 or 30 mg.kg(-1).d(-1) and CsA + MTX was better than that of MMF of 60 mg.kg(-1).d(-1) and CsA + MTX.</p>
Subject(s)
Animals , Female , Mice , Bone Marrow Transplantation , Cyclosporine , Pharmacology , Dose-Response Relationship, Drug , Drug Synergism , Graft Survival , Graft vs Host Disease , Pathology , Immunosuppressive Agents , Pharmacology , Methotrexate , Pharmacology , Mice, Inbred C57BL , Models, Animal , Mycophenolic Acid , Pharmacology , Time Factors , Transplantation, HomologousABSTRACT
<p><b>OBJECTIVE</b>To investigate the mechanisms of arsenic trioxide (As(2)O(3)) induced demethylation.</p><p><b>METHOD</b>Methylation of p15INK4B gene in MUTZ-1 cell was detected by PCR using a methylation specific primer (MSP), the expression of P15, DNA methyltransferase (DNMT) 1, DNMT3A and DNMT3B gene by RT-PCR, the As(2)O(3) induced growth inhibition of MUTZ-1 cell by MTT method.</p><p><b>RESULTS</b>P15 gene failed to express in MUTZ-1 cells after methylation. The expression was recovered after the cells exposed to As(2)O(3). As(2)O(3) could significantly down-regulate DNMT3A and DNMT3B but not DNMT1 gene on mRNA level in a dose dependent manner.</p><p><b>CONCLUSION</b>As(2)O(3) could activate the expression of p15 gene by demethylation or/and by inhibiting DNMT3A and DNMT3B gene.</p>
Subject(s)
Humans , Arsenicals , Pharmacology , Cell Line , Cell Proliferation , Cell Survival , Cyclin-Dependent Kinase Inhibitor p15 , Genetics , DNA (Cytosine-5-)-Methyltransferase 1 , DNA (Cytosine-5-)-Methyltransferases , Genetics , DNA Methylation , Gene Expression Regulation , Myelodysplastic Syndromes , Genetics , Pathology , Oxides , Pharmacology , RNA, Messenger , Genetics , Metabolism , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
AIM: To investigate the effects of transforming growth factor ?1 (TGF-?1) on murine-derived dendritic cells (DC). METHODS: Murine bone marrow cells were cultured with GM-CSF and TGF-?1 to develop TGF ?-DC. Then they were stimulated by lipopolysaccharide (LPS). Their phenotypes were assessed by flow cytometry (FCM). The allogeneic stimulating capacity of DC was measured by mixed lymphocyte reaction (MLR) using BrdU ELISA method. IL-12 p70 protein was detected by ELISA and the expressions of Toll like receptor 4 (TLR4) on DCs were measured by semi-quantitative RT-PCR and FCM. RESULTS: Compared to immature DC (imDC) cultured with GM-CSF alone, the expressions of CD_80, CD_86, I-A~b and CD_40 in TGF ?-DC were lower. The TGF ?-DC was resistant to maturation by LPS. Maturation resistance was evident from a failure to up-regulate CMs, to stimulate larger T cell proliferation and to increase secretion of IL-12 p70. Down-regulation of TLR4 expression on TGF ?-DC was also found. CONCLUSION: TGF-?1 inhibits the expression of co-stimulatory molecules on DC. It is resistant to maturation stimulus (LPS) and might be linked with TLR4 down-regulation.
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AIM: To explore the influence of CD47 molecules on the maturation and function of cultured dendritic cells (DCs). METHODS: Monocyte cell-derived DCs were propagated in granulocyte-macrophage colony-stimulating factor (GM-CSF) and lipopolysaccharide (LPS) plus interleukin (IL)-4, in the presence or absence of anti CD47 monoclonal antibodies (anti-CD47 mAbs). Flow cytometry was used to detect the cell surface phenotype. The concentration of IL-12P70 in supernatant was measured by ELISA technique. The antigen-presenting functions of DCs were determined in one-way mixed leukocyte reaction by Brdu-ELISA. Electrophoretic mobility shift assays (EMSA) was used to examine NF-?B activity. RESULTS: The anti-CD47 mAbs markedly suppressed the expression of CD80,CD86,CD83,CD1a,HLA-DR on the surface of DCs (P
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AIM: To explore the relationship between hypermethylation of p15INK4B gene and the pathogenesis of hematopoietic malignances. METHODS: The expression and methylation of p15INK4B gene and the expression of DNA methyltransferase genes (DNMTs) in bone marrow cells from 54 cases with hematopoietic malignances were detected by RT-PCR, Western blot, and methylation-specific PCR. RESULTS: The p15INK4B gene was methylated more often in high-risk myelodysplastic syndrome (MDS) patients, patients at blast phase of chronic myeloid leukemia (CML-BP) and acute leukemia patients than that in low-risk MDS patients (P
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AIM: To explore the effect of berbamine(BER) on apoptosis in K562 cells and its possible molecular mechanisms. METHODS: The apoptosis rate was measured by flow cytometry while electron microscopy and DNA electrophoresis were used to evaluate the characteristic changes of apoptosis, RT-PCR and Western blot were used to examine the expression levels of apoptosis related gene bcr/abl and BCR/ABL protein. RESULTS: By FCM, the apoptosis rate of K562 cells treated with 8.0 mg/L BER for 24 h and 72 h increased from (29.20?3.82)% to (61.77?4.35)% (P
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OBJECTIVE To analyze the risk factors,clinical characteristics and prevention countermeasures of(nosocomial) fungal infection in patients with acute leukemia.METHODS To adopt investigation way to review and analyze the clinical data of nosocomial fungal infection in 39 cases from 178 acute leukemia inpatients.RESULTS The(nosocomial) fungal infection rate was 21.91%.The nosocomial fungal infection rate in elderly group was(higher) than that of younger group,and in granulocytosis group(≥0.5?10~9/L) was higher than that of
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Effect of both granulocyte transfusions and antibiotics on the treatment of acute agranulocytosis with severe infections in 9 of 15 patients were reported. However, all 6 patients with agranulocytosis ind- uced by various drugs survived from sev- ere infections. It revealed that granulocyte count of each transfusion was more sign- ificant for the therapeutic effect.
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AIM: To investigate the infection of huma n embryo fibroblasts (HF) with CMV as well as the effect of CMV on ?-actin and mi crofilaments. METHODS: RT-PCR assay was used to detect the mRNA expression of CMV immediate early (IE) gene, ?-actin and GAPDH genes in HF cells infected wit h CMV. The morphological changes and microfilaments in infected cells were obser ved with transmission electron microscopy. RESULTS: The morphology of HF cells infected with CMV changed si gnificantly from fusiform shape to round shape. The mRNA expression of CMV immed iate early gene was detected. The increase in mRNA level of IE gene was parallel with the infected titer of CMV. However, t he expression level of ?-actin mRNA in HF cells infected with CMV was decreased compared with the uninfected cells, while the expression of GAPDH mRNA did not change. CMV particles were observed in the cells by electron microscope. Microfi laments were found ruptured and shortened after the infection of CMV. CONCLUSIONS: CMV was able to infect human embryo fibroblasts and replicated in the cells. Also the CMV infection affected the expressi on of ?-actin mRNA and the microfilaments.
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Dami) existed. The transcriptional levels of survivin gene in K562/ADR cell strain was 1.6-fold higher than that in K562 cell strain,but wasn't detected in peripheral blood mononuclear cells(PBMNCs).All-trans retinoic acid significantly decreased survivin mRNA expression levels. CONCLUSION: These data suggest that the survivin gene might be a potential therapeutics target due to the role in anti-apoptosis and drug resistance.
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AIM: To characterize the gene expression of sortilin on adipogenic and osteogenic differentiation in mesenchymal stem cells (MSCs) in vitro and explore its significance.METHODS: MSCs derived from human bone marrow were isolated and cultured in vitro ,then were stimulated in osteogenic medium and adipogenic medium,respectively. Osteopontin and lipoprotein lipase were detected by RT-PCR. Sortilin expression was analyzed by semiquantitative RT-PCR. RESULTS: 1.MSCs displayed the potential of differentiation into osteoblast and adipocyte. 2.Sortilin was upregulated one day after osteogenic induction and remained upregulated for a week. The expression of sortilin was significant increased on day 3( P 0.05).CONCLUSION: Sortilin may be useful to modulate the osteogenic differentiation and may not be necessary for adipocyte commitment in MSCs. The regulation of sortilin expression may provide new protocal and strategy for the treatment of osteoporosis and osteopenic disease.