To explore the effect of recombinant human interleukin 11(rhIL-11), granulocyte-colony stimulating factor (rhG-CSF) combined with cyclophosphamide (CTX) on mobilizing peripheral blood stem/progenitor cells in C57BL/6 mice. A total of 48 C57BL/6 mice were randomized into four groups. CTX (200mg/kg) was injected intraperitonealy on the first day (d0) in the treatment group. rhIL-11 alone or in combination with rhG-CSF was administered subcutaneously, beginning from 24 hours after CTX for 15 consecutive days in two other groups. The changes in peripheral blood white cell counts, platelet counts and the percentage of hematopoietic stem/progenitor cells of the mice were observed. It was shown that rhIL-11 or/and rhG-CSF could increase the numbers of WBC, PLT, CFU-GM, CFU-E, CFU-MK progenitor cells, and the percentage of CD34 positive cells in peripheral blood of myelosupressed mice. The results demonstrated that rhIL-11 alone or in combination with G-CSF and CTX could mobilize bone marrow haematopoietic stem/progenitor cells into peripheral blood.

As the first line of therapy for chronic myeloid leukemia, the outcome of imatinib is encouraging. The plasma concentration of imatinib is correlated with remission rate and survival, and affected by α1 acid glycoprotein(AGP) and those drugs which will change the activities of cytochrome P450(CYP3A4). It is conductive for the treatment of CML by monitoring the plasma concentration of imatinib.

Objective To set up a new real-time quantitative PCR method for the detection of minimal residual disease in chronic myeloid leukemia patients, and to assess the bcr/abl fusion gene expression in chronic myeloid leukemia patients before and after treatment with imatinib mesylate by real-time quantitative PCR method. Methods The bcr/abl fusion gene expression in 30 patients with bcr/abl-positive chronic myeloid leukemia was analyzed by using real-time quantitative reverse transcription PCR (RQ-PCR) method. The patients treated with imatinib in a dose of 400mg/d for 1 year and 2 years were also examined (8 cases for each). In 19 new patients the same study was also conducted. Results The real time quantitative PCR method could detect 10 copies in the test. The average bcr/abl expression levels in new patients or patients who had been treated with imatinib for 1 year and 2 years were 68.18%?26.67%, 0.16%?0.15% and 0.04%?0.02%, respectively. The average logarithm reduction values after treatment were 2.82 in the first year and 3.36 in the second year. In 25% of patients (4/16) negative FISH results could not be obtained, but it was much lower than that of before imatinib-treatment. When FISH became negative, RQ-PCR showed positive results. Conclusions RQ-PCR is a more sensitive technique in the detection of bcr/abl fusion gene than the FISH. It is an important way to monitor the tumor cell during the treatment with imatinib mesylate in chronic myeloid leukemia patients.

0 05). The PBSC with red blood cells(8～11ml) without elimination before infusion were harvested by CS 3000 Plus.The leve1 of blood group antigens and antibodies and bone marrow cells morphology were examined at different intervals after transplantation. No patient showed acute haemolysis in the ABO incompatible group. The median time of blood type transformation was 40 2?25 days. The median time for recovery of normal WBC and Plt in ABO incompatible groups was 10 days longer than that in the ABO compatible group. The number of patients with Hb

To investigate the role of vascular endothelial growth factor (VEGF) in occurrence and progression of acute myeloid leukemia (AML), enzume linked immunosorbent assay(ELISA) was used for detection of VEGF concentration in plasma from AML patients and normal bone marrow donors.The mean VEGF concentration in the plasma from refractory (558 90pg/ml) and non refractory (392 54pg/ml) AML patients was higher than that from normal donors (57 27pg/ml) and AML patients post Allo BMT (77 31pg/ml).There were significant differences between refractory and non refractory AML group. The baseline VEGF level (196 14pg/ml) of patients in complete remission (CR) after a median follow up of 6 months was significantly lower than that of patients with newly diagnosed or relapsed AML, but significantly higher than that of patients with Allo BMT AML and normal donors. Therefore, abnormal VEGF expession may play an important role in development of AML, and VEGF might be used to evaluate prognosis of AML.

Objective In the treatment of acute myeloid leukemia (AML-M2a), the first CCR (continuous complete remission) has been one of the most critical indicators to the prognosis of the patients. Using microarray approaches, gene expression profiles have been studied in patients with different CCR, in order to find out the genes relevant to the progresses of the AML. Methods Bone marrow mononuclear cells were collected and used as different experimental groups respectively. Group A composed of three AML patients with CCR<6 months, while group B composed of three AML patients with CCR>12 months. mRNAs were purified and labeled with Cy3 and Cy5 respectively, which were used to hybridize against the Agilent human 1B 60mer oligonucleotide microarrays. Results In the 20173 genes tested, 21 genes were found expressed differentially between these two groups. Of these differentially expressed genes, 10 genes were up-regulated while 11 genes were down-regulated in group A. Conclusion Through microarray studies, 21 genes including APP were found to be differentially expressed in AML patients whom were treated with standard chemotherapy. Theses genes can be early indicators for the diagnosis as well as prognosis of the refractory AML.

Echinoderms are deuterostome, and occupy the top taxonomic position of invertebrates, where is the evolutionary linkage between invertebrates and vertebrates.This is a critical phylogenetic vantage point to infer both the early evolution of bilaterian immunity and the underpinnings of the vertebrate adaptive immune system.The available published literatures on echinoderm immunological mechanisms are compiled and discussed here to understand its evolution process and the hot spots or directions in future investigations.Echinoderms have an innate immune system like vertebrates but their adaptive attributes have not been observed.Its immune responses are based on coelomocytes activity working in parallel with a variety of humoral factors that react directly with invading pathogens.Comparative studies of immune functions in echinoderms have demonstrated that there is a complement system in echinoderms that appears to have alternative pathway and lectin pathway but lack classical pathway and terminal pathway.The sea urchin innate immune system has a number of large gene families.Further investigations are needed to understand the unknown immune-associated genes, proteins, immune signaling pathways and effector molecules, and to know the origin, the underlying mechanisms and evolution of innate immune system.

Objective To explore the apoptosis effect induced by bortezomib combined with homoharringtonine or arsenious acid in HL-60 cell line and the mechanism.Methods Cell' s apoptosis was demonstrated by MTT assay and Hoechst33342 staining.Expression of bcl-2,Caspase-9,Caspase-3 and PARP protein was detected by Western blot.Results HL-60 cells' apoptosis could be induced by bortezomib,homoharringtonine and arsenious acid respectively.Proliferation inhibition of HL-60 cells could be enhanced significantly when treated by bortezomib combined with homoharringtonine or arsenious acid compared with treated by any of the three drugs alone (P ＜ 0.05).At the same time morphology shows the apoptosis induced by drugs combined is more obviously than by one drug.Western blot showed bcl-2 protein was down-regulated and Caspase-9,Caspase-3 and PARP proteins were all cleaved activation when cells were treated by 15 μmol/ L arsenious acid alone,but only cleaved activation of PARP and down-regulation of bcl-2 protein be detected when cells were treated with 30 nmol/L homoharringtonine alone,expression of Caspase-9 and Caspase-3 had no change compared with the control.The changes of associated proteins were paralleled with the cell' s apoptosis when treated with combined drugs.Conclusion HL-60 cells' apoptosis effect is inhanced significantly when bortezomib combined with homoharringtonine or arsenious acid.Arsenious acid and bortezomib can inhibit caspase signaling pathway and down-regulate the expression of bcl-2 protein together,but homoharringtonine and bortezomib can only down-regulate the expression of bcl-2 protein and induce cleaved activation of PARP together.

OBJECTIVE: To investigate the curative effect of combined transplantation of bone marrow and umbilical cord blood of same sibling in children with β-thalassemia major.METHODS: Eight thalassemia major patients undergoing combined transplantation of bone marrow and umbilical cord blood of same sibling aged from 4.0 to 7.5 years, 5 boys and 3 girls, were recruited at the Department of Pediatrics, Nanfang Hospital,Southem Medical University from January 2005 to March 2009. The patients were classified into three classes according to Pesarothalassamia classification, class Ⅰ to class Ⅱ 7 cases and class Ⅲ 1 case. Donors ranged 1-4 years received 10 μg/kg per day of subcutaneous granulocyte colony-stimulating factor (G-CSF) for 5 consecutive days. Bone marrow was harvested on the fifth day. Bone marrow and umbilical cord blood of the same sibling then were transfused into the patient.RESULTS: Recovery of hematopoiesis was gained in all patients 4 weeks following transplantation. Seven patients suffered from infection of different degree. Four patients developed mild venous occlusive disease. Two patients developed grade Ⅰ acute graft-versus-host disease (GVHD), and one developed grade Ⅰ chronic GVHD. Seven patients were alive and one died of pulmonary infection and heart failure 32 days following transplantation.CONCLUSION: Combined transplantation of granulocyte colony-stimulating factor primed bone marrow and umbilical cord blood of same sibling in children with β-thalassemia major is safe and effective with promising results. However, complications should be paid high attention following transplantation.

Objective To explore the effect of recombinant human granulocyte colony stimulating factor (rhG-CSF) mobilization on TH17/Treg cells and its impact on suppressor of cytokine signaling-3 (SOCS3) gene expression in CD4+ T cells in donors' peripheral blood.Method Sixteen donors were injected subcutaneously with rhG-CSF 5 μg/kg every day for 5 consecutive days for peripheral blood stem cells mobilization.At the first 0,3,5 day,the mononuclear cclls (MNCs) in peripheral blood or graft and serum specimens were taken.The CD4 + T cells in MNCs were sorted using immuno-magnetic beads.The ratio of TH 17 and Treg cells in MNCs,cytokines concentrations of IL-17A,IL-21,ID23 and TGFβ1 in serum,and SODC3 gene expression in CD4+ T cells were detected by using flow cytometry,ELISA,and reverse transcription real-time quantitative PCR (RT-qPCR),respectively.Results (1)The ratio of Th17 cells (CD3+ CD8 CD17+) and Treg cells (CD4+ CD25+ Foxp3+) in MNCs in peripheral blood and graft at the first 0,3 and 5 days after mobilization was (2.69 ± 0.81) ％,(0.91 ± 0.33) ％,(0.35 ± 0.12) ％,(0.21 ± 0.05) ％,and (0.56 ± 0.24) ％,(0.72 ± 0.22％),(1.59 ± 0.54) ％,(3.38 ± 0.52) ％,respectively,showing a significant declining and increasing trend respectively (P＜0.05); (2)The cytokine concentrations in serum at the first 0,3 and 5 days after mobilization were 7.33 ± 0.89,5.78 ± 1.03 and 3.32 ± 0.84 μg/L for IL-17A; 124.56 ± 15.18,117.12 ± 14.45 and 64.94 ± 11.25 μg/L for IL-21 ; 183.52 ± 59.35,280.49 ± 69.75 and 393.62 ± 57.25μg/L for TGF-β1 (P＜0.01) ; and 45.89 ± 6.95,46.25 ± 7.44 and 47.45 ± 10.75 μg/L for IL-23,respectively.The IL-17A and IL-21 concentrations showed significant declining trend,contrarily TGF-β1 with an increasing trend,while IL-23 concentration had no change.After rhG-CSF mobilization,the SOCS3 gene expression in CD4 + T cells of peripheral blood and graft at the first 0,3,5 days was gradually increased.Conclusion rhG-CSF suppresses Th17 cells and promotes regulatory T cells generation,meanwhile decreases IL-17A and IL-21 and elevates serum TGF-β1 concentrations,and contributes to CD4 + T cells differentiation to Tregs,probably by elevating SOSC3 gene expression in CD4+ T cells.