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Objective To explore the clinical and biological characteristics and treatment effect of acute myeloid leukemia (AML) in elderly patients. Methods The clinical data of 62 patients over 60 years old with AML were retrospectively analyzed, and they were compared with those of 60 younger adult patients (18-59 years old) at the same period. Results In elderly patients, the complete remission (CR) rate was 27. 7% and the overall effective rate was 44.7%, which were lower than those of younger adult patients (74.1% and 87.9 %, 1-espectively, P＜0.01). The early death rate was 19.0% and the mortality rate during the first two induction chemotherapy was 29. 3% in the elderly, which were higher than those of younger adult patients (3. 3% and 6.7%, 1-espectively,P＜0.01). The 27.4% of elderly patients were diagnosed as myelodysplastic syndrome (MDS)-transformed AML, which were more than that of younger adult patients (10.0%, P＜0.05), and they had lower CR rate than those without MDS (P＜0. 05). The 28.2% of elderly patients had lymphoid antigen positive AML and 71.8% of patients showed CD34+ which were higher than those of younger adult patients (8. 1% and 48.6%, respectively, P＜0. 05), and they had lower CR rate than those of lymphoid antigen negative and those of CD34- AML (P＜0. 05). Elderly patients had less favorable and more unfavorable karyotypes than younger adult patients (45.7 % and 15.4 %, P＜0. 05). Conclusions The elderly patients with AML have more unfavorable prognostic factors than younger adult patients. They have lower CR rate and higher mortality rate. There are many specialties in elderly patients and the treatment strategy should be made more individually.
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Objective To explore the immunophenotype of chronic lymphocytic leukemia (CLL). Methods The immunophenotypes of 31 patients with CLL were determined by immunocytometry. Results Among 31 cases with CLL, the positive expression rates of CD19,HLA-DR, CD5, CD23, CD20, CD22, CD38, FMC7 and CD10 were 100%, 96.8%, 90.3%,90.3%, 83.9%, 54.8%, 32.3%, 6.5% and 0.0%, respectively. Conclusions The imrnunophenotype analysis is very important for diagnosing CLL and it can early detect monoclonal B-cell lymphocytosis of CLL, which is the early phase of CLL, and provide early warning for patients.
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Objectives To evaluate the efficacy of rituximab combined with chemotherapy in the treatment of diffuse large B-cell lymphoma (DLBCL) and the relationship of clinical prognosis with the International Prognostic Index (IPI) by the using rituximab in autologous peripheral stem cell transplantation (APBSCT) for the patients of DLBCL. Methods 21 patients with DLBCL, 11 patients of them were at IPI low risk, and 3 patients were IPI at low intermediate risk, 3 patients were at IPI high intermediate risk, 4patients IPI high risk. Rituximab combined with CHOP regimen (cyclophosphamide, adriamycin, vincfistine and prednisone) was given for 4～8 courses. 5 patients received APBSCT. The mobilizing regimen was rituximab combined with cyclophosphamide(CTX) and etoposide(VP16). The conditioning regimen were CBV(CTX combined with VP16 and carmustine). Results In 21 patients, the complete response rate was 61.9 %,with overall response rate 90.5 %. 2-year progression free survival was (69.74±10.43)%. 2-year overall survival was (84.44:1:8.35) %. The complete response rate was 92.9 % and overall response rate was 100 % in the patients IPI≤2. The overall response rate was 71.4 % in the patients with IPI≥3. The complete response rate was higher in the patients with IPI≤ 2 (P＜0.01). The amount of mononuclear cells (M NC) in harvest were 7.34 (4.6～8.53)×108/kg. The CD+34 cells in harvest were 8.82 (2.1～10.34)×1O6/kg. The mean time of neutrephil recovering to 0.5×109/L after APBSCT was +9 day. The mean time of platelet recovering to 20×109/L after APBSCT was +12 day. The major adverse reaction were infusion related response (14.3 %) and hematological toxieities. Conclusion The efficacy of rituximab combined with chemotherapy in the treatment of DLBCL is effective, The complete response rate was higher in the patients with IPI≤2 than in the patients with IPI≥3.Using rituximab in mobilizing regimen, all patients had harvested enough CD+34 cells. Rituximab given at +1day did not affect the hematopoiesis reconstruction.
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Objective To evaluate the characteristics of immunophenotyping in adults with acute myeloid leukemia (AML) using muhi-eolor flow cytometry. Methods Immunophenotyping was performed by three color flow cytometry using CD45/SSC gating. Results In 126 patients with AML, the myeloid antigen of CD13, CD33 and CD117 was highly expressed. The positive rate was 86.4 %, 70.2 % and 90.4 %, respectively. The CD34 and HLA-DR were lowerly expressed as 63.5 % and 61.7 %, respectively. About 34.2 % of lymphoid-assoeiated antigen expression of all the AML patients. The lymphoid-associated antigens of CD7 and CD19 expression in patients with AML was 23.6 % and 2.3 %, respectively. Conclusion Multi-color flow cytometry is an important method for diagnosis and prognosis for AML.
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BACKGROUND: Cord blood stem cells are one of ideal target cells for gene therapy, but low gene transferring rate is the main difficulty at recent. Janus kinase tyrosine 2 (JAK2) plays an important role in self-renewing of cord blood stem/progenitor cell12s. Therefore, cord blood CD34+ cell line modified by target-amplified JAK2 genes has been developed yet by using gene regulating expression technique in order to overcome low transferring rate of cord blood genes.OBJECTIVE: To investigate the feasibility and reliability of a long-term amplified regulation for cord blood stem/progenitor cells mediated by transgene JAK2. SETTING: Department of Hematology, Beijing Hospital, Ministry of Health.MATERIALS: The experiment was carried out in the Laboratory of Hematological Department, Beijing Hospital, Ministry of Health from June 2003 to April 2006. Cord blood was derived from umbilical cord which was immediately cut from healthy, full-term and natural-parturition infants and was provided by Department of Obstetrics & Gynecology, Beijing Hospital. The experiment was approved by the local ethical committee, and informed consent was obtained from expectant mothers and their relatives for the use of cord blood cells. MiniMACS magnetic separation apparatus and immunomagnetic beads adsorbing CD34 single antibody were provided by Miltenyi Biotec Company, Germany; flow cytometer by FACScalibur, USA; recombinant human stem cell factor (rhSCF), Flt3 ligand (FL), human interleukin-6 (hIL-6), granulocyte macrophage colony stimulating factor (GM-CSF), granulocyte colony-stimulating factor (G-CSF) and thrombopoeitin (TPO) by PeproTec Company; nude mice of the SPF level by Animal Center of Beijing Medical University.METHODS: Retroviral vector MGI-F2JAK2, which was composed of functional catalytic domain of JAK2 genes and two site proteins (2xF36v, F2) combined with synthetic drug (AP20187) of target gene of small molecules, was constructed. AP20187 might specially combine with F36v to cause dimerization of JAK2 so as to activate signal conduction in cells. In addition, the vector included green fluorescence protein reporter gene, which was regarded as a label to detect proliferation. MiniMACS magnetic separation apparatus was used to purify and separate cord blood CD34+ cells. While, retrovirus supernatant including JAK2 was used to transfer cord blood CD34+ cells. After transduction, CD34+ cells were cultured with stem cell factor (SCF), Flt3 ligand, TPO and IL-6 and divided into control group (not adding AP20187) and experimental group (AP20187).MAIN OUTCOME MEASURES: ① Flow cytometer was used to detect percentage of green fluorescence protein reporter gene in the CD34+ cells and to determine gene transferring rate. ② Colony culture results of cord blood stem/progenitor cells after amplification. ③ Nude mice were given subcutaneous injection of ten-week cultured cord blood CD34+ cells at costa and neoplasia was observed after 30 days. RESULTS: ① Plentiful amplification of CD34+ cells was observed in both experimental group and control group. With the culture time passing by, positive rate of gel-filtered platelet of amplified CD34+ cells in the experimental group was gradually increased based on the basic level and more than 95% in the 11th week; however, positive rate of green fluorescence protein reporter gene in the control group was gradually decreased below the basic level and disappeared finally. ② Transgenic CD34+ cells in the experimental group still could generate brust forming unit-erythroid (BFU-E), colony-forming units granulocute/monocyte (CFU-GM) and multipotential hematopoietic progenitors (CFU-Mix); especially, CFU-GM was the main cell in hemopoietic progenitor cell (HPC). ③ Nude mice did not have neoplasia. CONCLUSION: Human cord blood CD34+ cells of transferring JAK2 genes may cooperate with other cytokines to amplify cord blood stem/progenitor cells in vitro for long. Therefore, this is potentially valuable for stem cells to treat some hereditary hematologic disease.
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AIM:To explore the feasibility and biological characterization of long-term regulated expansion of JAK2 transduced human CD34+ cord blood cells in vitro.METHODS: A retrovirus (RV) vector which contains JAK2 catalytic domain and two binding sites for a chemical inducer, dimerization (AP20187), was cloned (designated MGI-F2JAK2). CD34+cells were enriched from cord blood with a MiniMACS system. The purified CD34+cells were transfected with supernatant from the retrovirus packaging cell line that expressed JAK2. Following transduction, cells were expanded into four groups: AP20187 alone, FL alone, TPO, alone, AP20187+FL+TPO, respectively. The expanded cells were monitored by GFP expression, immunophenotyping, progenitor colony assay, karyotype analysis as well as tumorigenesis in nude mice. RESULTS: The purity of selected CD34+ cells was over 91% and gene transfer rate was 49.32%?6.21%. Only the group of AP20187 +FL+ TPO was obtained a significant sustained outgrowth of the transduced CD34+ cord blood cells. The percentage of GFP+ cells consistently produced a rise to the 90% peak level by the end of 8th week of culture. Flow cytometry analysis showed that the phenotype of the expanded cells was CD33+, CD61+ and Gly-A+ partial positive; CD38+ and HLA-DR+ strong positive, while CD2, CD7 and CD19 were almost negative. Colony assays performed in methycelluos, which can give rise to BFU-E, CFU-GM and CFU-Mix, the CFU-GM was predominantly in all colonies. The tumor was not observed in nude mice and the karyotype analysis was normal from expanded cells.CONCLUSION: The results demonstrate that AP20187-mediated activation of JAK2 signaling is capable of stimulating expansion JAK2 transduced CB CD34+ cells in combination with FL and TPO. This system may have applications for studies in signaling transduction, hematopoiesis, and for gene and cell therapy.