Azacitidine ; analogs & derivatives ; therapeutic use ; Clinical Trials, Phase III as Topic ; Humans ; Leukemia, Myeloid, Acute ; drug therapy ; Multicenter Studies as Topic

OBJECTIVE To determine the characterization, anti-tumor efficacy and pharmacokinetics of bufalin- loaded PEGylated liposomes compared with bufalin entity. METHODS Bufalin- loaded PEGylated liposomes and bufalin- loaded liposomes were prepared reproducibly with homogeneous particle size by the combination of thin film evaporation method and high pressure homogenization method. The particle size and zeta potential of the liposomes were determined by dynamic light scattering technique. The direct imaging of morphology of liposomes was charactered by transmission electron microscope. The content of bufalin in liposomes was analysed by HPLC method. The entrapment efficiency and the particle size was applied to assess the stability profile, after storage at 4℃ on day 0, 7, 15, 30 and 90. The in-vitro release behaviours of bufalin from liposomes were conducted using dialysis bag technique at 37℃. In-vitro cytotoxicity studies were carried out using MTT〔3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide〕assay on several kinds of tumor cell lines including SW620, PC-3, MDA-MB-231, A549, U251, U87 and HepG2. In-vivo pharmacokinetic study of bufalin liposomes was evaluated by HPLC method. RESULTS Their mean particle sizes were 127.6 nm and 155.0 nm, mean zeta potentials were 2.24 mV and - 18.5 mV, entrapment efficiencies were 76.31% and 78.40% , respectively. In- vitro release profile revealed that the release of bufalin in bufalin- loaded PEGylated liposomes was slower than that of bufalin-loaded liposomes. The cytotoxicity of blank liposomes has been found within acceptable range, whereas bufalin-loaded PEGylated liposomes showed enhanced cytotoxicity to U251 cells compared with bufalin entity. In-vivo pharmacokinetics indicated that bufalin-loaded PEGylated liposomes could extend eliminate half-life time of bufalin in plasma in rats. CONCLUSION The results suggested that bufalin-loaded PEGylated liposomes improved the solubility and increased the drug concentration in plasma.

The transcriptional coactivator MN1 has been identified as a gene overexpressed in certain types of human acute myeloid leukemia. Overexpression of this gene is associated with all inv (16) AML, retinoic acid-resistance, a worse prognosis as well as a shorter survival in AML patients with a normal karyotype. This article reviews the role of MN1 in acute myeloid leukemia including MN1 gene structure and action mechanism, MN1-TEL and AML with normal karyotype, MN1 and inv (16) AML, MN1 and retinoic ocid-resistance, and so on.
Humans ; Leukemia, Myeloid, Acute ; genetics ; Oncogene Proteins, Fusion ; genetics ; Transcription Factors ; genetics ; Tumor Suppressor Proteins ; genetics

The objective of this study was to evaluate the efficacy and toxicity of the fludarabine combination with high-dose cytarabine (Ara C), idarubicin and granulocyte colony-stimulating factor (G-CSF) (IDA-FLAG regimen) in treatment of refractory/relapsed acute leukemia (AL) patients. 4 patients were male aged from 32 to 44 years, consisted of 3 cases of acute myeloid leukaemia (AML) and 1 cases of acute lymphocytic leukaemia (ALL). All the patients were treated with idarubicin (10 - 12 mg/m(2)/d, days 1 to 3), fludarabine (50 mg/d, days 1 to 5), cytarabine (2 g/m(2)/d, days 1 to 5) and granulocyte colony-stimulating factor (G-CSF, 300 microg/d, days 0 to 5). The results showed that after one course of induction therapy, 4 patients all achieved complete remission (CR), in which 2 patients were in continuous CR after a follow-up of 3 and 4 months; 1 patient relapsed after 10 months and another one patient died of thrombotic thrombocytopenic purpura at 4 months after allogeneic peripheral blood stem cell transplantation. Myelosuppression and infections due to neutropenia were the most frequent adverse effects, severe nonhematologic toxicity and the early death were not observed in these patients. In conclusion, the IDA-FLAG regimen is effective in treatment of patients with refractory and relapsed AL, the adverse effects from this regimen were well tolerated by patients, which gains time for further treatment.
Adult ; Antineoplastic Combined Chemotherapy Protocols ; administration & dosage ; therapeutic use ; Cytarabine ; therapeutic use ; Granulocyte Colony-Stimulating Factor ; therapeutic use ; Humans ; Idarubicin ; therapeutic use ; Leukemia ; drug therapy ; Leukemia, Myeloid, Acute ; drug therapy ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; drug therapy ; Vidarabine ; analogs & derivatives ; therapeutic use

This study was purposed to investigate the mutational status of DNA methyltransferase (DNMT3a) gene and the clinical features of AML patients with DNMT3a mutations. Using PCR combined with directly sequencing, the somatic mutations of DNMT3a involving residue of amino acid 882 were detected in 77 AML patients. Furthermore, the clinical features of these patients were also studied. The results showed that the DNMT3a mutation were detected in 7 out of 59 patients with de novo AML (11.9%), which included 4 patients with DNMT3a R882C, 2 patients with DNMT3a R882H and 1 patient with DNMT3a Y874C. Morphology examination indicated that 2 patients were M(2), 1 patient was M(4) and 4 patients were M(5). Cytogenetic analysis revealed that karyotype in 5 out of 7 patients with DNMT3a mutation were normal. In total of 27 patients with normal karyotype 5 patients (22.7%) were found harboring DNMT3a mutation, while no DNMT3a mutation was found in 21 patients with abnormal karyotype. The mutation rate in patients with positive CEBPA was obviously higher than that in patients with negative CEBPA (p = 0.002). Immunophenotype analysis showed that 4 patients (4/7, 57.1%) with DNMT3a mutation expressed lymphoid antigens including CD4 or/and CD7. There were no statistical significance in age, gender, blast cells of bone marrow, white blood cell and platelet counts, hemoglobin level, ratio of CR, mutations of FLT3-ITD, NPM1 and c-kit between patients with DNMT3a mutation and patients with wild DNMT3a (p > 0.05). It is concluded that the DNMT3a mutations are more prevalent in AML patients with normal karyotype accompanying with positive NPM1 and/or CEBPA mutation, the role of DNMT3a mutation in AML prognosis needs to be further studied.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; CCAAT-Enhancer-Binding Proteins ; genetics ; Child ; DNA (Cytosine-5-)-Methyltransferases ; genetics ; Female ; Humans ; Leukemia, Myeloid, Acute ; genetics ; Male ; Middle Aged ; Mutation ; Nuclear Proteins ; genetics ; Young Adult

OBJECTIVETo investigate the value of multiplex fluorescence in situ hybridization (FISH) in the detection of complex karyotypic abnormalities of acute myeloid leukemia (AML).

METHODSMultiplex FISH was used in combination with conventional cytogenetics (CC) and interphase FISH to study 14 cases of AML with complex karyotypic abnormalities.

RESULTSIn the 14 cases of AML studied, conventional cytogenetics detected 23 numerical and 56 structural chromosome abnormalities. Among them 4 gained whole chromosome and 4 lost whole chromosome which were confirmed by multiplex FISH. Twelve chromosome losses detected by CC were revised as derivative chromosomes resulted from various structural aberrations, and 26 derivative and 19 marker chromosomes were characterized precisely by multiplex FISH. Most of them were resulted from unbalanced translocations, including 2 complex 8; 21 translocations, which have not been reported previously: t (8; 21), der (8) t (8; 21) (8pter --> 8q22::21q22 --> 21qter), der (21) t (8; 21; 8) (8qter --> 8q22:: 21p13 --> 21q22::8q22 --> 8qter) and t (21; 8; 18; 1), der (8) t (8; 21) (8pter --> 8q22:: 21q22 --> 21qter), der (21) t (21; 8; 18; 1) (21p13 --> 21q22?::8q22 --> 8q24 ?:: 18??::1q??q??). The complex karyotypic abnormalities involved nearly all chromosomes, of which the chromosomes 17, 7 and 5 were more involved than the rest.

CONCLUSIONMultiplex FISH in combination with conventional cytogenetics may characterize the complex chromosomal abnormalities more precisely. Introduction of this technique to the study of AML with complex chromosomal abnormalities is warranted.

Acute Disease ; Adolescent ; Adult ; Female ; Humans ; Leukemia, Myeloid ; genetics ; pathology ; Male ; Middle Aged ; Spectral Karyotyping ; methods ; Translocation, Genetic ; Young Adult

To investigate the effect of velcade on multiple myeloma cell line U266 apoptosis and its mechanism, cell viability was estimated by trypan blue dye exclusion. Annexin-V, mitochondrial transmembrane potential (delta psi m) and reactive oxygen species (ROS) labeled by DCFHDA were examined by flow cytometry, the expression of bcl-2 mRNA was detected by semi-quantitative RT-PCR. The results showed that the velcade inhibited the growth of U266 cells and reduced cell viability accompanied by appearance of morphologic characteristics of apoptosis. Velcade at 50 nmol/L increased Annexin V positivity and fluorescence intensity of DCF because of ROS generation while it decreased the delta psi m of U266 cells. Expression of anti-apoptotic gene bcl-2 mRNA also decreased. It is concluded that velcade inhibited the growth and reduce cell viability of U266 cells. Velcade can induce U266 cells apoptosis by intrinsic cell apoptotic pathway.
Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Boronic Acids ; pharmacology ; Bortezomib ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Humans ; Multiple Myeloma ; metabolism ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; biosynthesis ; genetics ; Pyrazines ; pharmacology ; RNA, Messenger ; biosynthesis ; genetics

This study was aimed to investigate the sensitivity and clinical application value of interphase-dual-color and dual-fusion fluorescence in situ hybridization (DD-FISH). The minimal residual disease (MRD) in 19 patients with chronic myelogenous leukemia (CML) after allogeneic hematopoietic stem cell transplantation (allo-HSCT) was detected by DD-FISH, and the detected results were compared with those of conventional cytogenetics (CC) and reverse transcription-polymerase chain reaction (RT-PCR). The samples were collected from bone marrow or peripheral blood or smears of bone marrow. The results indicated that 14 out of 19 patients achieved and maintained continuous complete molecular remission after transplantation. In these patients, CC assay displayed normal donor karyotype, result of RT-PCR was negative, complete donor chimerism was detected after 2 months of transplantation, result of DD-FISH was negative, average time of the follow-up survey was 11.25 months, MRD did not increase. Results of CC and RT-PCR in 1 patient showed negative, while FISH of sex chromosome showed mixed chimerism, result of DD-FISH was positive, MRD did not increase, no therapy was given for this patient, clinical state of patient was stable. Three patients with hematological relapse demonstrated obvious increase of MRD detected by DD-FISH and sex FISH, result of RT-PCR was found positive in them, but the abnormal result of CC was observed only in 1 patient. After donor lymphocyte infusion and imatinib mesylate treatment, these 3 patients achieved cytogenetic remission again, results of DD-FISH, CC and PCR were negative in them. DD-FISH, CC and PCR in bone marrow and peripheral blood from one patient with extramedullary relapse revealed negative results, and the complete chimerism was found in this patient. It is concluded that interphase-dual-color and dual-fusion fluorescence in situ hybridization is a more reliably sensitive and practicable method for monitoring MRD in patients with CML after allo-HSCT, and can be used in detection of chromosome sample and blood or bone marrow smears. Dynamic detection of bcr/abl fusion gene level by FISH may predict disease changes and guide individual therapy.
Adolescent ; Adult ; Female ; Fusion Proteins, bcr-abl ; genetics ; Hematopoietic Stem Cell Transplantation ; Humans ; In Situ Hybridization, Fluorescence ; methods ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; therapy ; Male ; Middle Aged

This study was aimed to establish the technique of fluorescence immunophenotyping and interphase cytogenetics as a tool for the investigation of neoplasms (FICTION) used to smear of bone marrow, so as to develop a new technique for detection of the molecular cytogenetic abnormalities in multiple myeloma (MM). By using the bone marrow smear as the carrier and the anti-CD138 antibody linked with FITC, direct fluorescence staining was applied to mark plasma cells (PCs) and differences were compared in the proportion of both PCs marked by fluorescence staining and PCs detected in morphology. At the same time, the chromosome 8 centromere probe was used in interphase fluorescence in situ hybridization (I-FISH) for detection of the chromosome 8 abnormalities in PCs marked by fluorescence staining. The results showed that there was no significant difference between the proportions of both PCs marked by fluorescence staining and PCs detected in morphology on smear (p>0.05). 4 out of 9 patients (44%) had the chromosome 8 abnormalities, including 3 cases with -8 (33%) and one case with +8 (11%). It is concluded that the FICTION technique on the basis of bone marrow smear is characterized by convenience, specificity and accuracy. Therefore, it can be used for molecular cytogenetic research in MM.
Aged ; Bone Marrow ; pathology ; Chromosome Aberrations ; Female ; Humans ; Immunophenotyping ; In Situ Hybridization, Fluorescence ; methods ; Male ; Middle Aged ; Multiple Myeloma ; genetics ; pathology

This study was purposed to investigate immune reconstitution at 12 months after allogeneic peripheral blood stem cell transplantation (all-PBSCT) and its relation with the influencing factors such as age, HLA compatibility, graft versus host disease and viral infection. The T lymphocyte subgroups (CD3(+), CD4(+), CD8(+)), B lymphocyte (CD19(+)) and NK (CD16(+)CD56(+)) cells in peripheral blood and serum immunoglobulin concentrations (IgG, IgA and IgM) of 37 patients were analyzed by flow cytometry and scatter turbidimetry, respectively at 1, 3, 6 and 12 months after transplantation. The results showed that CD3(+) cell percentage was (47.5 +/- 23.2)% at 1 month, (75.1 +/- 6.4)% at 3 months, (69.7 +/- 12)% at 6 months and (71.7 +/- 4.2)% at 12 months. CD4(+) cell percentage was (13.3 +/- 6.4)% at 1 month, (20.2 +/- 11.4)% at 3 months, (46.9 +/- 10.3)% at 6 months and (29.1 +/- 18.7)% at 12 months. CD8(+) cell percentage was (43.1 +/- 23.2)% at 1 month, (42.6 +/- 16.9)% at 3 months, (69.7 +/- 12)% at 6 months and (47 +/- 5.6)% at 12 months. CD16(+)56(+) cell percentage was (14.4 +/- 8.4)% at 1 month, (15.9 +/- 7.6)% at 3 months, (14.7 +/- 6.6)% at 6 months and (13.6 +/- 3.4)% at 12 months. CD19(+) cell percentage was (6.4 +/- 5.6)% at 1 month, (11.7 +/- 2.4)% at 3 months, (13.3 +/- 7.3)% at 6 months and (16.7 +/- 5.7)% at 12 months. The serum concentration of IgA was (0.37 +/- 0.14) g/L at 1 month, (0.28 +/- 0.21) g/L at 3 months, (0.42 +/- 0.18) g/L at 6 months and (0.53 +/- 0.34) g/L at 12 months. The serum concentration of IgG was (12.7 +/- 3.8) g/L at 1 month, (16.3 +/- 5.2) g/L at 3 months, (14.3 +/- 6.2) g/L at 6 months and (15.4 +/- 6.9) g/L at 12 months. The serum concentration of IgM was (0.56 +/- 0.24) g/L at 1 month, (0.64 +/- 0.16) g/L at 3 months, (1.1 +/- 0.35) g/L at 6 months and (1.2 +/- 0.28) g/L at 12 months. There were no significant differences between percentage of T lymphocyte subgroups in peripheral blood and serum immunoglobulin concentrations of the patients > or = 45 years old and the patients < 45 years old. The CD19(+) cell percentage of the patients with chronic GVHD at 12 month was less than that of the other ones at 12 months after transplantation. CD4(+) and CD19(+) cell percentage recovery in the patients of haploidentical transplantation was later than that in patients of HLA complete identical transplantation. The CD4(+)/CD8(+) cell ratio and CD4(+) cell percentage of those patients infected with herpes zoster were significantly lower than those without herpes zoster. It is concluded that the CD3(+) cell percentage begins to recover at 3 months after allo-PBSCT. CD4(+) cell percentage begins to recover at 6 months after allo-PBSCT. CD8(+) cell percentage begins to recover at 1 month after allo-PBSCT. B cell percentage recovers at 3 to 6 months after allo-PBSCT. NK cell percentage recovers at 1 to 3 months after allo-PBSCT. The serum concentration of IgG recovers to normal at 1 month after transplantation which is associated with routine infusion of immunoglobulin. The concentration of IgM gradually recovers to normal at 3 months after transplantation. The concentration of IgA does not recover to normal at 12 months after transplantation. The function of B cells recovers slowly in patients with cGVHD. The CD4(+) cell absolute value and CD4(+)/CD8(+) ratio significantly decrease in patients with herpes zoster.
Adolescent ; Adult ; Female ; Graft Survival ; Graft vs Host Disease ; Humans ; Killer Cells, Natural ; immunology ; Lymphocyte Count ; Male ; Middle Aged ; Peripheral Blood Stem Cell Transplantation ; Postoperative Period ; T-Lymphocyte Subsets ; immunology ; Young Adult