Hematologic Diseases ; Humans ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells

Objective To investigate the relationship between the expression of DNA methyltransferases(DNMT)and clinical prognosis in children with acute leukemia(AL).Methods The mRNA expressions of DNMT1,DNMT3A,DNMT3B,p15,mdrl were measured in 56 AL children and 20 normal controls by semi-quantitative reverse transcriptionpolymerse chain reaction.Results In 56 cases of children with AL,the positive rate of DNMT1 was 73.2%(41/56);the positive rate of DNMT3A was 67.9%(38 /56);the positive rate of DNMT3B was 64.3%(36/56).Thirty-one cases showed positive expressions of the 3 DNMT simultaneously,4 cases with negative expressionss imultaneously,21 cases with at least 1 positive expression of the DNMT,positive rate of p15 was 19.6%(11/56);positive rate of mdrl was 28.6%(16/56),all 3 simultaneous expressions of the 3 DNMT in AL children were significantly higher than those in normal controls(P

OBJECTIVETo investigate the mechanism of demethylation therapy of leukemia by 5-aza-2'-deoxycytidine (5-aza-CdR).

METHODSBy using MTT test, NBT reduction reaction and DNA agarose gel electrophoresis, changes in proliferation, differentiation and apoptosis were observed in K562, HL-60 and fresh leukemia cells after treated with 5-aza-CdR. The mRNA expressions of DNMTs, p15, p53 and bcl-2 were measured by RT-PCR. The status of p15(INK4B) gene methylation was examined by methylation-specific PCR (MSP-PCR).

RESULTSThe growth inhibition of K562, HL-60 and fresh leukemia cells displayed a dose and time-dependent manner after treated by 5-aza-CdR. The differentiation-inducing ability on HL-60 cells was obvious at 0.5 micromol/L of 5-aza-CdR. The up-regulation of p15 mRNA and p53 mRNA expression and down-regulation of bcl-2 mRNA expression were obvious as compared with the control, but the DNMTs expression was not significantly different from the control. The methylation status of p15 gene in fresh leukemia cells decreased gradually with increasing concentration of 5-aza-CdR.

CONCLUSIONThe proliferation of leukemia cells was obviously inhibited by 5-aza-CdR, its mechanism maybe related to the up-regulation of p15 and p53 genes and down-regulation of bcl-2 gene. The decrease of p15 gene methylation was associated with the competitive inhibition of 5-aza-CdR.

Antimetabolites, Antineoplastic ; pharmacology ; Apoptosis ; drug effects ; Azacitidine ; analogs & derivatives ; pharmacology ; Cell Cycle Proteins ; genetics ; metabolism ; Cell Proliferation ; drug effects ; Cell Transformation, Neoplastic ; drug effects ; Cyclin-Dependent Kinase Inhibitor p15 ; DNA Methylation ; drug effects ; DNA Modification Methylases ; metabolism ; Gene Expression Regulation, Neoplastic ; drug effects ; HL-60 Cells ; Humans ; K562 Cells ; RNA, Messenger ; biosynthesis ; genetics ; Tumor Suppressor Proteins ; genetics ; metabolism

OBJECTIVETo explore the biological function of vascular endothelial cells from hypertrophic scar, and to analyze the relationship between them.

METHODSThe samples from human hypertrophic scar and normal skin tissue were harvested for histological examination. Then vascular endothelial cells were purified and isolated from the samples, and the level of transforming growth factor (TGF) beta1, platelet derived growth factor (PDGF), endothelin1 (ET)-1, fibroblast growth factor (FGF)2 and vascular endothelial growth factor (VEGF) were determined in a single cell with ELISA.

RESULTSFew capillary vessels were observed in normal skin under microscope, while an increased number of them were present in hypertrophic scar, with slender, tortuous in morphology and even occluded. The diameter of blood capillary in hypertrophic scar was tiny under electron microscope, and the exfoliation of endothelial cells was observed. The levels of TGF-beta1, PDGF, ET-1, bFGF and VEGF from vascular endothelial cells from hypertrophic scar were 60 +/- 8, 30 +/- 4, 0.12 +/- 0.03, 52 +/- 5, 18.1 +/- 1.2 microg/cell, respectively, which were obviously lower than those in normal skin (P < 0.05).

CONCLUSIONThe biological function of vascular endothelial cells was attenuated in the hypertrophic scar, which mightbe the result of the production of large amounts of collagen in the scar tissue, as well as hypoxia.

Adolescent ; Adult ; Cells, Cultured ; Cicatrix, Hypertrophic ; metabolism ; pathology ; Collagen ; metabolism ; Endothelial Cells ; metabolism ; Endothelin-1 ; metabolism ; Female ; Fibroblast Growth Factor 2 ; metabolism ; Humans ; Male ; Platelet-Derived Growth Factor ; metabolism ; Skin ; blood supply ; Transforming Growth Factor beta1 ; metabolism ; Vascular Endothelial Growth Factor A ; metabolism ; Wound Healing ; Young Adult

BACKGROUNDLenalidomide has emerged as an important treatment for patients with multiple myeloma (MM). However, its role in the management of MM is still controversial and requires further clarification. The aim of this study was to evaluate efficacy and safety of lenalidomide for MM using a meta-analysis.

METHODSWe searched the electronic databases including: PubMed, EMBASE and the Cochrane Center Register of Controlled Trials. Seven randomized clinical trials were identified, which included a total of 2357 patients with MM who received lenalidomide-containing, noncontaining lenalidomide regimens or placebo as induction therapy or maintenance therapy. The outcomes included overall response (OR) rate, complete response (CR) rate, 3-year progression-free survival (PFS) rate, 3-year overall survival (OS) rate, and different types of treatment-related adverse events. We calculated the risk ratios (RRs) as well as their 95% confidence intervals of these outcomes and pooled the results using RevMan 5.2 software.

RESULTSFor patients with previously untreated MM, OR rate and CR rate was significantly higher in lenalidomide-containing group than the control group. For relapsed or refractory MM patients, lenalidomide-containing regimens significantly improved the OR rate, CR rate, 3-year PFS rate and 3-year OS rate. With regard to MM patients after autologous stem cell transplantation, lenalidomide maintenance therapy significantly improved 3-year PFS rate but did not result in improved 3-year OS rate. In terms of toxicities, lenalidomide therapy has a higher rate of Grade 3-4 grade cytopenias, infection, deep-vein thrombosis, and diarrhea. Furthermore, the incidence of second primary malignancies was significantly higher in the lenalidomide group.

CONCLUSIONSThe lenalidomide-containing regimens as induction therapy clearly increased response rates and improved intervals of survival with acceptable toxicity rates for patients with MM. However, when physicians choose to use the lenalidomide as maintenance therapy, whether the benefits outweigh the risks should be taken into account.

Angiogenesis Inhibitors ; adverse effects ; therapeutic use ; Humans ; Multiple Myeloma ; drug therapy ; Randomized Controlled Trials as Topic ; Thalidomide ; adverse effects ; analogs & derivatives ; therapeutic use ; Treatment Outcome

To improve the recognition of immunoglobulin D multiple myeloma and explore its clinical feature and laboratory examination characteristics, so as to reduce the the missed diagnosis and misdiagnosis, a case of IgD multiple myeloma (MM) with myelofibrosis and bone marrow necrosis is reported. The clinical feature, treatment and prognosis of IgDlambda MM were discussed. Immunoglobulin D multiple myeloma is a rare disease and predominantly occurs in young male patients, which shows an aggressive clinical course with poor response to conventional treatment and unfavorable prognosis. Immunoglobulin D multiple myeloma was usually misdiagnosed as a light chain type multiple myeloma by using routine laboratory examination. Immunoglobulin D monoclonal protein is not easy to be detected owing to its low protein level, resulting in missed diagnosis. Immunofixation electrophoresis is highly sensitive and specific for diagnosis of IgD MM, can enhance accuracy of diagnosis for this rare disease.
Adult ; Diagnosis, Differential ; Humans ; Immunoglobulin D ; blood ; Immunoglobulin lambda-Chains ; blood ; Male ; Multiple Myeloma ; blood ; complications ; diagnosis ; Primary Myelofibrosis ; blood ; complications ; diagnosis

OBJECTIVETo investigate the effect of ERCC1 gene on the repair capability of damaged DNA in lung cancer A549 cells induced by benzo[a]pyrene.

METHODSRecombinant plasmid expressing ERCC1 antisense RNA was constructed and transfected into A549 cells by Lipofectin reagent. The stable-transfected cell colonies were selected by hygromycin. Cell viability was determined by the MTT assay. The level of ERCC1 mRNA was measured by Northern Blot analysis. Single cell gel electrophoresis assay was applied to determine the cellular DNA damage and fifty cells for each group were counted.

RESULTSSeven positive colonies expressing ERCC1 antisense RNA were screened. There was no growth rate difference between the antisense-transfected cells and the parental cells. The endogenous mRNA level in transfected colonies decreased in varied degrees, i.e. 12% approximately 86% of that of the parental cells in Northern Blot assay. After 24 h treatment of 10 micro mol/l benzo[a]pyrene, the repair capability for DNA damage in transfected colonies was reduced to 29% approximately 71% of that of the parental cells. Also, a statistically significant correlation was observed between expression of ERCC1 mRNA and repair capability (r = 0.84).

CONCLUSIONAntisense ERCC1 RNA decreased the repair capability for damaged DNA in lung cancer cells induced by benzo[a]pyrene.

Benzo(a)pyrene ; toxicity ; Cell Line, Tumor ; DNA Damage ; drug effects ; DNA Repair ; drug effects ; DNA-Binding Proteins ; genetics ; metabolism ; pharmacology ; Endonucleases ; genetics ; metabolism ; pharmacology ; Humans ; Lung Neoplasms ; pathology ; Plasmids ; RNA, Antisense ; pharmacology ; RNA, Messenger ; metabolism ; Repressor Proteins ; Transfection

To investigate the relationship between the expression of DNMT and clinical prognosis in adult patients with acute leukemia (AL), the mRNA expressions of DNMT, p15(INK4B), mdr1 were measured in 72 AL patients and 20 normal controls by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR); the ratio of p15 CpG land methylation was measured in 56 AL patients and 14 normal controls by methylation-specific PCR (MSP-PCR). The results showed that all three DNMT mRNA expressions in AL patients were significantly higher than that in normal controls (P < 0.01). When the internal control was changed into PCNA, a kind of cell proliferation marker gene, the difference still showed a statistic significance. All three DNMT genes were significantly expressed and positively correlated with AL patients, showing high synergistic expression, and there was a negative correlation between the levels of p15, mdr1 gene expression and DNMT. The complete remission (CR) rate in AL patients with the positive expression of all DNMT genes was significantly higher than that of AL patients with partially positive or negative expression (P < 0.01) of DNMT genes. In 56 AL patients, the P15I(NK4B) was completely methylated in 55.4% (31 of 56), partly methylated in 21.4% (12 of 56) and all 14 cases of normal controls were not methylated. It is concluded that DNMT genes are abnormally high expressed in adult AL patients, which lead to methylation-silence of tumor suppressor genes by CpG land hypermethylation, the AL patients with high expression of DNMT are more sensitive to chemotherapy, which may be a good prognostic factor for AL patients.
ATP Binding Cassette Transporter, Sub-Family B ; ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; Acute Disease ; Adult ; Cyclin-Dependent Kinase Inhibitor p15 ; genetics ; DNA (Cytosine-5-)-Methyltransferase 1 ; DNA (Cytosine-5-)-Methyltransferases ; biosynthesis ; genetics ; Female ; Gene Expression Regulation, Leukemic ; Humans ; Leukemia ; genetics ; pathology ; Male ; Prognosis ; Proliferating Cell Nuclear Antigen ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

This study was purposed to investigate the expression of c-fes gene in leukemia patients and its clinical significance. The expression of c-fes mRNA in bone marrow cells from 121 cases of acute and chronic leukemia patients, and the expression of c-fes mRNA in peripheral blood mononuclear cells of 20 normal persons were detected by real time-quantitative reverse transcription polymerase chain reaction (RQ-PCR). The results showed that the level of c-fes mRNA in AML patients was higher than that in normal controls [(48.017 +/- 57.170) x 10(-3) vs (0.152 +/- 0.398) x 10(-3)] (p < 0.0001); but there was no significant differences of level of c-fes mRNA between samples of ALL and normal controls(0.047 +/- 0.068) x 10(-3) vs(0.152 +/- 0.398) x 10(-3) (p>0.05); the level of c-fes mRNA in CML patients was higher than that in normal persons (21.605 +/- 24.818) x 10(-3) vs (0.152 +/- 0.398) x 10(-3) (p < 0.0001). The positive expression rate of c-fes gene in CML-CP patients (80%) was higher than that in CML-AP patients (66.7%) and CML-BP (28.6%) patients. In AML patients, c-fes gene was expressed higher in M(2) (80.77%) and M(3) (92.86%) patients. The remission rate of AML (except M(3))patients who had expression of c-fes gene was 81.08%, which was higher than that of patients with no expression of c-fes gene (40.00%). It is concluded that c-fes gene expression was found in myeloid leukemias, whereas low or no expression in lymphocytic leukemias. The differentiation of myelocytic cells may be related to c-fes gene. All AML (except M(3))patients with high level of c-fes mRNA may get good prognosis.
Adult ; Case-Control Studies ; Female ; Gene Expression ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; Leukemia, Myeloid ; genetics ; Leukemia, Myeloid, Acute ; genetics ; Male ; Prognosis ; Proto-Oncogene Proteins c-fes ; genetics ; RNA, Messenger ; genetics

Adult ; Bone Plates ; utilization ; Clavicle ; surgery ; Female ; Humans ; Internal Fixators ; trends ; Male ; Middle Aged ; Young Adult