Cord blood is the main source of stem cell outside of bone marrow and peripheral blood, and cord blood transplantation, especially unrelated cord blood transplantation (UCBT) has been widely used. The major challenges in UCBT are delayed engraftment and graft rejection. Many approaches have been investigated to enhance collection of HSC and HPC in cord blood units,sinduding use of double unit UCBT, adopt reduced intensity conditioning (RIC) approach, amplification of cord blood cells in vivo or ex vivo, and co-infusion with a haploidentical T cell depleted graft or mesenchymal stem cells(MSC),or injecting cord blood cells directly into the bone marrow.Those approaches may greatly increase the clinical use of cord blood cells for transplantation.

Objective To establish a rapid assay for the determination of methotrexate (MTX) in serum.Method The assay was based on the ultraviolet absorbance of methotrexate at 306 nm. The separation of the drug was done by high performance capillary electrophoresis (HPCE). The bare fused-silica capillary was 60 cm in total length, 50.5 cm in efficient length and 75?m in diameter. The voltage of 25 kV was applied. The running buffer was 75 mmol/L phosphate, pH7.4. The performance of methodology was evaluated.Result The complete separation of MTX was achieved within 10 min. The linearity of the assay was from 1.1 ?mol/L to 1100.0 ?mol/L. The minimal detection limit was 0.55 ?mol/L.The recovery of MTX was from 88.2% to 98.2%. Within-run precision was 4.2% and between-run precision was ~5.4%. Conclusion The result indicated that the method was an effective method for clinical and scientific research with advantages of rapidity, simplicity and accuracy.

Objective:To investigate the relationship between levels of plasma thrombopoietin(TPO) and thrombocytopenia.Methods:Sixty-eight thrombocytopenia with different causes were injected with rhIL-11 domestically made, by 25 ?g/(kg?d) for 10 days continuously. The plasma TPO levels were determined by quantitative sandwich enzyme immunoassay technique in thrombocytopenic patients between before and after treatment.Results:(1)The serum TPO controls of patients with thrombocytopenia after acute leukemia(AL) chemotherapy were lower than normal controls. The plasma TPO levels in aplastic anemia(AA) and myelodysplastic syndrome(MDS) patients were all elevated compared to those in normal controls.The serum TPO levels in patients with idiopathic thrombocytopenic purpura(ITP) and liver cirrhosis had no difference with the normals. Bone marrow megakaryocytes(MK) count in after chemotherapy AL and AA patients were lower than those with normal serum TPO levels controls. (2)In all above patients used interleukin-11(rhIL-11), their TPO levels were close to normal in efficiency patients. On the contrary, their TPO did not change dramatic. Those react to rhIL-11, their TPO levels were inversely correlated with platelet count.Conclusion:Testing serum TPO levels would be helpful for distinguish the thrombocytopenia with different causes, and provides theorical support to patients to use rhIL-11 reasonably.

Purpose　The aim is to obtain the cDNA sequence of encoding extramembrane human FL gene with high level expression in E.coli. Methods　The primers were designed based on the known FL cDNA sequence. The total RNA was isolated from fetal liver cells , and then RT-PCR was performed. The fragment was cloned into pUC-18T vector, and further sequenced by automatic sequence analyzer. The gene was inserted into GST fusion expression vector between BamH Ⅰ and EcoR Ⅰ sites. The recombinant plasmid was transformed into E.coli strain DH5 α and induced with 1mmol/L IPTG.Results　The 546bp DNA fragment was amplified by RT-PCR method from fetal liver cells and its sequence was identical to the published sequence encoding human FL. The expressed fusion protein, with molecular weight of about 22kD, was about 10% of the total bacteria protein by SDS-PAGE and densitometry analysis.Conclusion　cDNA was cloned successfully. This study provided a basis for the further fundamental research and clinical application of FL.

Objective To develop an assay with polyclonal antibodies against a fragment derived from human albumin for determination of a peptide in urine, and to provide an early diagnostic tool for GVHD. Methods A small peptide composed of 16 amino acids (LVRYTKKVPQVSTPTL) was synthesized artificially. The immunogen was then diluted into 100 μg/kg body weight of rabbit. Subcutaneous injection in the immune animals was performed on both sides of spine and groin with 2.5 ml antigen suspension for three times, in order to prepare the polyclonal antibodies. The peptide antigen was then labeled with fluorescein isothiocyanate (FITC), and detected by LIF-CE-IA with the pre-prepared antibodies. Results The titer of serum. The migration time of the labeled antigen was 5.93 min, while the migration time of antigen-antibody complex was 6.47 min. The linear range of the method was 16 to 512 ng/ml, and the minimum detection limit was 10 ng/ml. Conclusions The polyclonal antibodies against the peptide antigen was isolated successfully, which possessed high titer and specificity. These results indicated that the assay was simple and rapid and could be applied for the early diagnosis of patient with GVHD.

Objective To analyze the fusion genes derived from 29 types of chromosome structural aberrations in leukemia patients,and the significances on the MICM typing,risk grouping,and minimal residual disease(MRD)monitoring of leukemia.Methods The bone ulan-ow or blood samples from 141 leukemia patients were analyzed with a novel multiplex nested RT-PCR.In addition.chromosomal karyotypes were investigated in some patients.Results Of the 141 leukemic samples,66(46.8%)carried 13 types of MLL/AF6,MLL/AF9,dupMLL MLI/ENL,CBFβ/MYH11 and TLS,ERG.Fusion genes were positive in 27 of 57 ALL patients(47.4 q%),and 33 of 78 AML patients(42.3%),respectively.In these ALL or AML patients,7 or 6 chromosome structural aberrations were found. Conclusion This multiplex nested RT-PCR reaction could screen 29 types of chromosome structural aberrations at the same time. It may be helpful for the diagnosis, risk grouping,prognosis evaluation and the detection of minimal residual diseases after chemotherapy and bone marrow transplantation in these leukemia patients.

Objective To investigate the efficacy and safety of posaconazole in the prophylaxis or salvage treatment of invasive fungal disease (IFD) in patients with hematological diseases.Methods 25 patients with hematological diseases receiving posaconazole treatment from Feb 2014 to Feb 2015 were analyzed retrospectively.The patients' average age was 32.6 years old (16-64 years old).18 patients received posaconazole for IFD prophylaxis, and 7 patients for IFD salvage treatment.Results 18 patients receiving posaconazole for IFD prophylaxis had no clinical manifestation of IFD (that is no case of breakthrough fungal infection) during treatment or in the 12 weeks after treatment, and the average prophylaxis period was 21 d (14-35 d).Among 7 patients receiving posaconazole for IFD salvage treatment, 6 patients were effective, including 4 cases cured and 2 cases effective.There were no obviously side effects of posaconazole in these patients.Conclusion Posaconazole has good clinical response for IFD in the hematologic diseases patients whether in prophylaxis or in salvage treatment.

Objective To analyze the treatment efficacy and safety of a modified GMALL protocol for adult acute lymphoblastic leukemia (ALL). Methods Data of 37 patients with newly diagnosed adult ALL treated with a modified GMALL protocol from January 2005 to December 2009 were retrospectively analyzed,and compared with that of 44 patients treated with an in-house conventional protocol at the same period.Results The complete remission (CR) rate was 89.2 %(33/37) treated with modified GMALL protocol. The cumulative overall survival (OS) rates at 1 year, 2 years, 3 years and 4 years were 77.5 %, 48.0 %, 40.0 %and 40.0 %, respectively. The main adverse events were grade 3 or grade 4 hematological toxicities and infections which were easily managed, and the treatment-related mortality rate was low. The OS of modified GMALL protocol was superior to that of the conventional protocol. Conclusion The modified GMALL protocol has a satisfying effect and the adverse events can be tolerated for adult ALL, so its clinical application can be encouraged.

[Objective] To retrospectively analyze the outcome of unrelated umbilical cord blood transplantation in the treatment of aggressive-phase chronic myeloid leukemia.[Methods] Fourteen consecutive patients with aggressive-phase chronic myeloid leukemia were treated with unrelated umbilical cord blood transplantation,thirteen patients were treated with myeloablative unrelated CBT and one patients were treated with nonmyeloablative unrelated CBT.All patients received standard cyclosporine A (CsA) and mycophenolate mofetil(MMF) as a graft-versus-host disease (GVHD) prophylaxis.[Results] 14 patients were all successfully engrafted.The median times for their neutrophil returning to ≥0.5×109/L and for platelet returning to ≥20×109/L were 22.8 days and 37.8 days,respectively.Acute GVHD occurred in 10 of 13 evaluable patients.The grading of acute GVHD was gradeⅡ-Ⅳin 6 patients(46.2 ％).Chronic GVHD occurred in 7 of 11 evaluable patients(63.6％).Relapse occurred in 2 of 15 patients,lextramedullary relapse was included.9 of 14 patients were alive and event-free after CBT.The probability of OS rate at 5 years was 64.3 ％,the probability of DFS rate at 5 years was 5,7.1％.[Conclusion]Unrelated umbilical cord blood transplantation is effective in the treatment of aggressive-phase chronic myeloid leukemia.

Objective To analyze the clinical and biological features of mixed acute leukemia(MAL).Methods Bone marrow specimens of 38 MAL patients were evaluated to prove the diagnosis and the classification by morphoiogic,immunologic examinations.These patients were treated with protocols suitable for both acute myeloid leukemia(AML)and acute lymphoblastic leukemia(ALL).Results All MAL patients had a leukemia syndrome.Morphologically,the subtypes of M1,M2 and M5 were predominant in AML,as L2 Was in ALL.Immunologically,coexpression of myeloid and B lineage associated antigens was predominant,about 68.4%;cytogenetically,Ph chromosome was observed in 33.3%(5/15)of MAL patients,and immunophenotype was B-M;1 Ph chromosome(+)MAL patient,fusion gene bcr-abl 190(+)and immunophenotype was B-M.In 38 cases,32 patients received chemotherapy.The complete remission rate was 28.1%(9/32).CR of.normal karyotype was significantly higher than that of abnormal ones.Conclusion Patients with MAL have unique biological features and the complete remission rate was low and the prognosis was poor.