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.

Objective To investigate the research quality of present medical graduates and propose suggestions for improvement.Method A self-designed questionnaire entitled Investigating Questionnaire for Research Quality Analysis of Medical Graduates in Peking Union Medical College Hospital was distributed to the clinical and academic medical graduates in the hospital from July to August 2015.Among 276 collected questionnaires,270 were validated as effective.SPSS 18.0 software was used to statistically describe the result and to perform t test on different subgroups.Results The research quality average of the surveyed hospital was 10.28.Results revealed that in regard of the three aspects of research quality,research consciousness ranked first,followed by the research ability,while scientific spirit was the weakest.Academic medical graduates showed significantly higher scores than clinical graduates in terms of total research quality and every single aspect (P values less than 0.05).Conclusions To better cultivate the academic leaders in medical research,it is necessary to strengthen the research training for the graduates,practice their scientific thinking especially the clinical graduates,reinforce tutors' guidance,and promote communication and collaboration.

Adolescent ; Adult ; Automobile Driving ; China ; Electrocardiography ; Female ; Heart ; physiopathology ; Humans ; Male ; Middle Aged ; Noise, Occupational ; adverse effects ; Occupational Exposure ; adverse effects ; Stress, Psychological ; etiology ; physiopathology ; Vibration ; adverse effects

OBJECTIVE To investigate the incidence and pathogens of infection in 1659 consecutive cases in single center hematological unit.METHODS The incidence,pathogen,and outcome of infection in 1659 hospitalized patients with hematological disorders from 1999 to 2006 were retrospectively analyzed.RESULTS The overall incidence of infection was 24.4% according to the person-times of hospitalization,which included 22.1% of nosocomial infection and 2.3% of community acquired infection.Most of the pathogenic bacteria of the nosocomial infection were Gram-negative.The most common bacteria in the sputum samples included Enterobacter cloacae(19.3%) and Pseudomonas aeruginosa(14.8%).The most common bacteria in the blood samples were coagulase negative Staphylococcus(CNS,39.3%),the next was Pseudomonas aeruginosa and Escherichia coli.There were 4.21% bacteria resistant to most of antibiotics in nosocomial infection.There were 114 fungi isolated.Candida albicans was accounted for 35.1%.The mortality due to nosocomial infection was 7.4%.CONCLUSIONS The patients in hematology ward are susceptible to infection.The pathogens of nosocomial infection are most likely G-bacteria.Some bacteria are resistant to almost all antibiotics.The incidence of fungal infection is increasing in the near 8 years.

Objective To explore the karyotype distribution in elderly patients with acute leukemia (AL) and compare the prognostic characteristics of karyotype by age grouping.Methods Chromosomal karyotypes were analyzed in 215 cases with AL using the short-term culture of bone marrow cells and G-banding technique.Results There were 202 cases with enough mitosis for analysis and 149 cases(73.8％)with abnormal clone in 215 patients with AL.The rates of abnormal clone were 73.0％ (27/37),74.4％(64/86) and 73.4％ (58/79) in patients aged ≤30,31-59 and ≥60 years,respectively,and no difference were found among age groups (P=0.982).Among 171 patients with acute myeloid leukemia (AML) with detected mitosis,there were 41 better-risk cases (24.0 ％) with most frequent aberration of t(15;17) accounting for 65.9 ％,80 intermediate-risk cases (46.8 ％ ) with principal of normal karyotype accounting for 53.8 ％,and 50 poor-risk cases (29.2 ％)with complex karyotype occupied by 84.0％.The karyotype percentage of better-risk,intermediaterisk and poor-risk were 50.0％,36.4％ and 13.6％ in patients aged ≤30 years,24.3％,48.7％ and 27.0％ in aged 31-59 years,and 16.0％,48.0％ and 36.0％ in aged ≥ 60 years,respectively.The rate of better-risk karyotype was higher in patients aged ≤30 years than the other two groups (P=0.021and P=0.001) and the ratio of poor-risk karyotype higher in patients aged ≥ 60 years than in patients aged ≤30 years (P=0.046).Among 29 patients with acute lymphoblastic leukemia (ALL),10 cases had poor-risk and 19 cases had intermediate-risk karyotype.Conclusions Karyotype analysis provides an important basis for risk assessment and the rate of poor-risk karyotype may increase with the ageing in patients with AML.

Objective To explore the cytogenetic characteristics of acute myeloid leukemia(AML) patients.Methods The karyotype analysis was performed in 178 AML using the short-term culture of bone marrow cell and G-banding technique.Results Among the 178 patients,171 had enough metaphases for analysis and 128(74.9%)had clonal karyotypic abnormalities.Twenty-seven patients were secondary to myelodysplastic syndrome (MDS-AML),with 25 (92.6%) patients carrying clonal karyotypic abnormalities.Among the remaining 144 patients of de novo AML,103(71.5%)had clonal karyotypic abnormalities.The rate of abnormal clonal karyotype was higher in MDS-AML than that of de novo AML (P=0.021).Among the 171 patients,41(24.0%)were in favorable risk group,80(46.8%)in intermediate risk group and 50(29.2%)in adverse risk group.t(15;17)was the most common chromosomal aberration.The maiority intermediate risk chromosomal aberration was;normal karyotype.The most common cytogenetic abnormality among adverse group was a complex karyotype.Adverse cytogenetic aberrations,such as -5/5q-,-7/7q-,frequently occurred in conjunction with one another as part of a complex karyotype.Totally 75 patients were 60 years or older,among them,16.0%were in favorable risk group,48.0%in intermediate risk group and 36.0%in adverse risk group.Among 96 younger patients,30.2%were in favorable risk group.45.8%in intermediate risk group and 24.0%in adverse risk group.The rate of favorable risk chromosomal aberration was lower in elder patients than in younger(P=0.03 1).The rate of adverse risk chromosomal aberration and the rate of monosomal karyotype were higher in MDSAML than in de novo AML patients(P<0.001).Conclusions The most common favorable,intermediate and adverse chromosomal aberrations were t(15;17),normal karyotype and complex karyotype respectively.The karyotype was poor in MDS-AML and elder AML patients.

Objective To explore the clinical characteristics,therapy reactions and prognosis of the elderly patients with idiopathic thrombocytopenic purpura(ITP). Methods A total of 43elderly ITP patients(age≥60 years old)including 16 men and 27 women were reviewed and further followed up for 1 month to 15 years. Results Until June 2007,35 elderly ITP patients survived,platelet counts were sustained(30-50)×109/L in 7 cases,but no significant bleeding was found.Thirty-six patients had adrenocorticosteroid therapy first, 25 patients were sensitive to adrenocorticosteroid therapy,4 patients underwent splenectomy,and 3 patients achieved a normal platelet count. Immunosuppressive agents(vinscristine,cyclophosphamide, azathioprine and Cyclosporin A)treatments were held in 5 6 case-times,Cyclosporin A and azathioprine were more effective than vinscristine and cyclophosphamide.The refractory rate was 13.9%.One patient progressed to monoclonal gammopathy of unknown significance and 1 to lymphoma.Eight patients died.1 patient died of brain bleeding after trauma,3 patients died of malignant neoplasm,4 patients died of heart failure induced by infection. Conclusions The clinical features of elderly ITP patients are atypical.the mortal bleeding in them was rare,treatment strategy should be individualized tO each elderly patient.

Objective To explore the outcome of monoclonal gammopathy of undetermined significance (MGUS). Methods The data from 14 MGUS patients in our hospital including clinical features, outcome and change of M protein concentration were analyzed retrospectively. Results The MGUS didn't have the clinical manifestations of multiple myeloma (MM), the time of outcome from MGUS to MM was about 4-20 years (mean time, 10 years). The most types of MM were IgA and IgG, 6 cases were IgA type, 6 cases were IgG type and 2 cases were light chain type. The concentration of immune globulin in general showed an upward trend year by year. A few showed fold lines ascend. Conclusions The elevated monoclonal immunoglobulin may develop into MM after many years. We must follow up frequently to avoid error diagnosis and missed diagnosis.

To explore the epidemiological character of Methicillin-resistant Stapkylococcus aureus (MRSA) by the phenotyping and genotyping motheds and to investgate the source, transmission, and the spread of nosocomial MRSA infection, consequently, reducing the nosocomial infection of MRSA. In this study, 19 MRSA strains were isolated from patients and environment in a hospital in two months. Patterns of resistantce against 16 antimicrobial agents and pulsed-field gel electrophoresis ( PFGE) of these strains were analyzed to find the relationship among those isolates Clustering analysis was made from the patterns. Some isolates with high homology was found in 19 MRSA, 11 of them belong to type A, and 8 of them belong to the same subtype A1. They were endemic in burn ward, oncological ward and ICU. In addition, 4 isolates were clustered into group B, all found in the same ward of burn unit Thus, our results indicated a outbreak of MRSA ( A type) in this hospital and the potential prevalence of MRSA (B type) , which might be mediated by health care stuff. It is essential to enhance the infection control implementation and to utilize the PFGE genotyping system for the real-time surveillance of MRSA.

Objective To compare the saccharometabolism with the pancreatic islets functions and insulin resistance index in children with severe stress. Methods Thirty children with severe stress and 30 healthy children in control group were tested. The levels of fasting blood glucose (FBG), fasting insulin (FINS) and fasting C - peptide (FCP) were detected by radioimmunoassay respectively and insulin sensitivity index (ISI), insulin resistance index (IR) and fasting blood cell function index (FBCI) were calculated statistically. Results There were significant differences between the children with severe stress and the normal controls in the levels of FINS, FCP and FBG,(all P0.05). Conclusion There is insulin resistance with the significant decrease in the insulin sensitivity index and significant increase in insulin resistance index in the children with severe stress, which may cause the disorder in glucose metabolism in children with severe stress.