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 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 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 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 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.

BACKGROUND:The elderly hip fracture patients with chronic obstructive pulmonary disease (COPD) have a higher postoperative mortality than those only with hip fractures. In recent years, it has become an issue of concerns. Because the mechanism is unknown, however, there are no effective clinical interventions for these patients. OBJECTIVE:To observe the effect of bone marrow stromal stem cel s (BMSCs) on the level of pulmonary inflammation in aged rats with chronic obstructive pulmonary disease (COPD) after hip fractures. METHODS:Thirty male Sprague-Dawley rats, 12 months old, were exposed to smoking for 4 months and randomized into three groups, smoking, smoking+hip fracture+normal saline, smoking+hip fracture+BMSCs groups. Animal models of hip fracture were made in the latter two groups. Twenty-four hours after hip fracture, the lower lobe and peripheral blood samples were taken from al rats in the three groups, to evaluate the pathological changes of lung tissue and detect levels of inflammatory factors in the lung tissue and peripheral blood. RESULTS AND CONCLUSION:After closed hip fracture, aged COPD rats exhibited inflammatory cel infiltration, mucus secretion, airway stenosis or occlusion;the levels of interleukin-6, tumor necrosis factor-αand interleukin-10 in the lung tissue and peripheral blood were increased. After intravenous injection of BMSCs, the pathological changes of the lung tissue were reduced, and the levels of pro-inflammatory factors, tumor necrosis factor-αand interleukin-6, decreased, but the level of anti-inflammatory factor interleukin-10 further increased, which were significantly different from those in the normal saline group (P<0.05). These findings indicate that BMSCs can relieve acute lung injury in aged COPD rats with hip fractures.

Objective To study the syndrome of inappropriate ADH secretion(SIADH)after all ogeneic hematopoietic stem cell transplantation(allo-HSCT)and the possible etiology.Methods SIADH was occurred in one patient with acute lymphoblastic leukemia who received secondary haploidentical stem cell transplantation after graft failure following umbilical cord blood transplantation.Results An 11-year-old boy was diagnosed as high-risk acute lymphoblastic leukemia,and the transplantation of unrelated double umbilical cord blood was performed for the first time,however,there was no engraftment at 28 days after the first transplantation.Secondary haploidentical stem cell transplantation was administered at day 33 after the first transplantion.The bone marrow and the peripheral blood stem cells from the patient's father transplantation was adopted.Pre-engraftment syndrome was developed in the patient at day 7 after secondary haploidentical HSCT followed by severe hyponatraemia(lowest serum sodium 115.8 mmol/L),natriuresis,hypo-osmolality of plasma,and twitch at day 26 after allo-HSCT.Then SIADH was diagnosed.The clinical condition was improved after restriction of water and administration of hypertonic saline,and eventually SIADH was controlled completely.By now,the patient had lived free of disease more than one year,with a normal diet and serum sodium in normal ranges.Conclusions SIADH after allo-HSCT is a rare fatal acute complication of central nervous system,whic h are probably associated with numerous transplant-related causes.Early accurate diagnosis and treatment promptly are great importance.

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.