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 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.

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 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.

Objective To analyze the clinical features and the effect of therapy on cutaneous emphysema of chest wall and/or pneumomediastinum complicated in severe acute triphosgene poisoning patients.Methods Among 81 triphosgene poisoning patients,5 complicated with cutaneousemphysema of chest wall and/or pneumomediastinum were analyzed in respect of the clinical data including age,gender,arterial blood gas (ABG),modes of mechanical ventilation support and so on.Results Five patients consisting of 3 males and 2 females,aged (23.20 ± 5.17) years,were complicated with cutaneous emphysema of chest wall and/or pneumomediastium with a prevalence rate of 0.06％.Of them,4 were alleviated completely and 1 died of acute respiratory distress syndrome (ARDS).There was no significant difference in arterial blood gas analysis (ABG) between patients with cutaneousemphysema and/or pneumomediastinum and patients without ( P ＞ 0.05 ).Conclusions Triphosgene-induced acute lung injury treated with mechanical ventilation support with high PEEP is highly suggested as high risk factor for the formation of cutaneous emphysema of chest wall and/or pneumomediastinum in severe acute Triphosgene poisoning patients.It is very important to set the PEEP level of mechanical ventilation support as low as possible for avoidance of alveolar rupture.

Objective To retrospectively analyze the engraftment, transplant-related complications and survival after unrelated cord blood transplantation (UCBT) in patients with hematologic malignancies. Methods Fifty consecutive patients with hematological malignancies (median age, 19 years; median weight, 53 kg) were treated with UCBT in single center from April 2000 to August 2009. Thirty-nine patients were high-risk or refractory. Double UCB grafts were used for 26 patients, while single UCB graft for 24 patients. Myeloablative conditioning was given to 45 cases and non-myeloablative regimens to 5 cases. All patients were given a combination of cyclosporin A (CsA) and mycophenolate mofetil (MMF) for graft-versus-host disease (GVHD) prophylaxis. Results The median total nucleated cell (TNC) dose was 4.0 (range, 1.95-16.24)×10~7 TNC/ kginfused, and CD34~+ cell dose was 2.74(range, 0.67-29.28)×10~5/kginfused. Forty-two of 50 patients acquired engraftment with implantation rate being 86%. The median time to engraftment (absolute neutrophil count>500/mm~3 and platelets 20 000/L) was 19 and 34 days. The cumulative incidence of neutrophil engraftment by day 42 was 86.3%(95% confidence interval [CI] 0.769-0.957); the cumulative incidence of platelets engraftment by day 120 was 72.3% (95% CI 0.620-0.821). Twenty cases developed acute GVHD, and the incidence of acute GVHD of grades Ⅲ/Ⅳ by day 100 was 7.1%. The incidence of chronic GVHD within 2 years was 17.4%. During a median follow-up period of 22 months (range 4-116), Overall 6-month, 1-year and 2-year survival rate was 66.2%(95% CI 0.590-0.734), 57.4%(95% CI 0.496-0.652), 54.2%(95% CI 0.462-0.622), respectively. For the patients with non-advanced hemotologic malignancies, 6-month, 1-year and 2-year survival rate was 73.2% (95% CI 0.659-0.805), 66.1% (95% CI 0.579-0.743), and 62.2% (95% CI 0.542-0.682) respectively. Five cases relapsed. The cumulative incidence of relapse within 2 years was 16.2% (95% CI 0.099-0.225). Twenty-one cases died mainly due to infection. Conclusion UCBT could be safely and effectively used for adult patients with hematologic malignancies.

Objective To investigate the safety and clinical outcome of total body irradiation (TBI) and the real-time in vivo dosimetry with semiconductor dosimeter in hematogenous stem cell transplantation (HSCT). Methods Fifty-seven patients requiring HSCT were treated with TBI. The TBI was given with the semi-sitting or standing position or lateralcumbent posture, using 6 MV X-ray beams and opposed parallel fields technique (two or four fields, AP/PA fields) in a single fraction or multiple fractions. The real-time in vivo dosimetry was performed with six diodes positioned on the surface of patients to adjust the dose homo-geneity of the midplane using the different thickness lead sheets. Results Mild to moderate nausea, vomi-ting and swollen parotid occurred in 41 patients after TBI, which were relieved after allopathy therapy. No radiation-induced interstitial pneumonia was observed. All patients fulfilled the HSCT. The homogeneity of relative dose (normalize to umbilicus dose) in the different positions accorded with the requirement of the prescription dose. Conclusions The opposed parallel radiation, with the semi-sitting or standing positions or lateralcumbent posture, combined with the real-time in vivo dosimetry with semiconductor dosimeter is an effective and safe technique for TBI.