Objective To study whether IL-1β, a catabolic factor of cartilage metabolism, induces premature senescence of articular chondrocytes, and whether caveolin-1 mediates IL-1β-induced cellular senescence. Methods Cultured human articular chondrocytes were stimulated with 10 ng/ml IL-1β. Cellular senescent phenotypes were analyzed by cellular morphology, cell growth arrest (flow cytometry), telomere erosion (Southern blotting), life span (population doublings), and specific senescence-associated β-galac-tosidase (SA-β-Gal) activity. Expression level of caveolin-1 was modulated by anti-sense oligunucleotide or transfection of caveolin-1 gene. Caveolin-1 protein was analyzed by Western blotting. Results Incubation of chondrocytes with IL-1β markedly increase the percentage of cells in G0/G1 phase and reduce the percentage in S phase. Single stimulation with IL-1β enables chondrocytes to become big and flat, and SA-β-Gal activity in chondrocytes is enhanced. Repeated stimulation with IL-Iβ resulted in accelerats erosion of mean telomere length, and shortens life span. Down-regulation of caveolin-1 with anti-sense oligonucleotide significantly inhibits the features of chondrocytes senescence induced by IL-1β. In contrast, caveolin-1 overexpreasion enhanced SA-β-Gal activity in the chondrocytes. Conclusion IL-1β induces features of stress-induced premature senescence and telemere-dependent replicative senescence of articular chondrocytes, which is mediated by caveolin-1. These data suggest that IL-1β induces premature senescence of articular chondro-cytes by upregulation of caveolin-1, which facilitates the development of osteoarthritis.

Objective: To observe effects of Yuzhang Decoction on in vitro proliferation of bone marrow CD_(34) cells in the patient of aplastic anemia (CAA) and its mechanism. Methods: To adopt monoclone antibody-immune magnetic beads separation system (MACS) to separate and purify CD_(34) cells in bone marrow of the patient of CAA, and make cell culture, applying ~3H-TdR incorporation to detect effect of Yuzhang Decoction on proliferation of CD_(34) cells of bone marrow. Results: CD_(34) cells of bone marrow showed concentration-dependent proliferation in the Yuzhang Decoction group with significantly differences (P

Objective To evaluate the characteristics of immunophenotyping in adults with acute myeloid leukemia (AML) using muhi-eolor flow cytometry. Methods Immunophenotyping was performed by three color flow cytometry using CD45/SSC gating. Results In 126 patients with AML, the myeloid antigen of CD13, CD33 and CD117 was highly expressed. The positive rate was 86.4 %, 70.2 % and 90.4 %, respectively. The CD34 and HLA-DR were lowerly expressed as 63.5 % and 61.7 %, respectively. About 34.2 % of lymphoid-assoeiated antigen expression of all the AML patients. The lymphoid-associated antigens of CD7 and CD19 expression in patients with AML was 23.6 % and 2.3 %, respectively. Conclusion Multi-color flow cytometry is an important method for diagnosis and prognosis for AML.

BACKGROUND: Cord blood stem cells are one of ideal target cells for gene therapy, but low gene transferring rate is the main difficulty at recent. Janus kinase tyrosine 2 (JAK2) plays an important role in self-renewing of cord blood stem/progenitor cell12s. Therefore, cord blood CD34+ cell line modified by target-amplified JAK2 genes has been developed yet by using gene regulating expression technique in order to overcome low transferring rate of cord blood genes.OBJECTIVE: To investigate the feasibility and reliability of a long-term amplified regulation for cord blood stem/progenitor cells mediated by transgene JAK2. SETTING: Department of Hematology, Beijing Hospital, Ministry of Health.MATERIALS: The experiment was carried out in the Laboratory of Hematological Department, Beijing Hospital, Ministry of Health from June 2003 to April 2006. Cord blood was derived from umbilical cord which was immediately cut from healthy, full-term and natural-parturition infants and was provided by Department of Obstetrics & Gynecology, Beijing Hospital. The experiment was approved by the local ethical committee, and informed consent was obtained from expectant mothers and their relatives for the use of cord blood cells. MiniMACS magnetic separation apparatus and immunomagnetic beads adsorbing CD34 single antibody were provided by Miltenyi Biotec Company, Germany; flow cytometer by FACScalibur, USA; recombinant human stem cell factor (rhSCF), Flt3 ligand (FL), human interleukin-6 (hIL-6), granulocyte macrophage colony stimulating factor (GM-CSF), granulocyte colony-stimulating factor (G-CSF) and thrombopoeitin (TPO) by PeproTec Company; nude mice of the SPF level by Animal Center of Beijing Medical University.METHODS: Retroviral vector MGI-F2JAK2, which was composed of functional catalytic domain of JAK2 genes and two site proteins (2xF36v, F2) combined with synthetic drug (AP20187) of target gene of small molecules, was constructed. AP20187 might specially combine with F36v to cause dimerization of JAK2 so as to activate signal conduction in cells. In addition, the vector included green fluorescence protein reporter gene, which was regarded as a label to detect proliferation. MiniMACS magnetic separation apparatus was used to purify and separate cord blood CD34+ cells. While, retrovirus supernatant including JAK2 was used to transfer cord blood CD34+ cells. After transduction, CD34+ cells were cultured with stem cell factor (SCF), Flt3 ligand, TPO and IL-6 and divided into control group (not adding AP20187) and experimental group (AP20187).MAIN OUTCOME MEASURES: ① Flow cytometer was used to detect percentage of green fluorescence protein reporter gene in the CD34+ cells and to determine gene transferring rate. ② Colony culture results of cord blood stem/progenitor cells after amplification. ③ Nude mice were given subcutaneous injection of ten-week cultured cord blood CD34+ cells at costa and neoplasia was observed after 30 days. RESULTS: ① Plentiful amplification of CD34+ cells was observed in both experimental group and control group. With the culture time passing by, positive rate of gel-filtered platelet of amplified CD34+ cells in the experimental group was gradually increased based on the basic level and more than 95% in the 11th week; however, positive rate of green fluorescence protein reporter gene in the control group was gradually decreased below the basic level and disappeared finally. ② Transgenic CD34+ cells in the experimental group still could generate brust forming unit-erythroid (BFU-E), colony-forming units granulocute/monocyte (CFU-GM) and multipotential hematopoietic progenitors (CFU-Mix); especially, CFU-GM was the main cell in hemopoietic progenitor cell (HPC). ③ Nude mice did not have neoplasia. CONCLUSION: Human cord blood CD34+ cells of transferring JAK2 genes may cooperate with other cytokines to amplify cord blood stem/progenitor cells in vitro for long. Therefore, this is potentially valuable for stem cells to treat some hereditary hematologic disease.

Objective To explore the immunophenotype of chronic lymphocytic leukemia (CLL). Methods The immunophenotypes of 31 patients with CLL were determined by immunocytometry. Results Among 31 cases with CLL, the positive expression rates of CD19,HLA-DR, CD5, CD23, CD20, CD22, CD38, FMC7 and CD10 were 100%, 96.8%, 90.3%,90.3%, 83.9%, 54.8%, 32.3%, 6.5% and 0.0%, respectively. Conclusions The imrnunophenotype analysis is very important for diagnosing CLL and it can early detect monoclonal B-cell lymphocytosis of CLL, which is the early phase of CLL, and provide early warning for patients.

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 After studying the change rule of the blood brain barrier(BBB) permeability, to provide the imaging evidence of radiotherapy effect evaluation and choose optimal chance of chemotherapy during whole brain radiotherapy (WBRT) in patients with brain metastasis by 99mTc DTPA brain SPECT. Methods 19 patients with brain metastasis who undergone WBRT by 60Co were studied. All patients underwent 99mTc DTPA brain SPECT before, during WBRT. The total counts radio of the tumor and contralateral brain (T/N) before and during WBRT(20 Gy) was calculated. Results 1) The average T/N before WBRT and during WBRT in 32 tumors was 1.892?1.094 and 2.167?1.637 respectively. (t =2.433,P

Objective To comprehensively evaluate the effect of the program of treatment and assistance to advanced schis-tosomiasis patients in Hunan Province from 2004 to 2013. Methods The fund investment of the program,the profits of hospi-tals and the improvement of the patients’health were investigated by data collection and questionnaire survey. The evaluation index system of treatment and assistance to advanced schistosomiasis in Hunan Province was constructed by the Delphi method and analytic hierarchy process,and the program was assessed comprehensively. Results The evaluation index system includ-ing 6 primary indices and 33 secondary indices was established. Among all the primary indices,the score of the treatment and assistance(22.25)was the highest,and that of the satisfaction assessment(8.15)was the lowest,and the score of the compre-hensive assessment was 87.06. The average cure rate of the patients was 13.08%from 2004 to 2013. More than 60%of the pa-tients’disease condition got better,and nearly 70%of the patients’psychological condition improved,and more than 70%of patients’self-help ability and social contact improved,as well as family happiness increased. In addition,the annual average cost for caretakers decreased by 2000 Yuan,and the profits of all the fixed-point hospitals for treatment and assistance in-creased. Conclusion The effectiveness and efficiency of the treatment and assistance to advanced schistosomiasis patients in Hunan Province is obvious,and the government should continuously invest in the program.

ischemia reperfusion model .DADLE might have a protective effect on lung tissues of ALI in rats .