Human vascular endothelial cells(VEC)were isolated from umbilicalyeins by trypsin digestion.Spleen cells from BALB/c mice having been im-munized with the isolated endothelial cells were fused with the mousemyeloma cell line SP2/0.After screening and subcloning,three differentmonoclonal antibodies(EC1,EC2,EC3)against human umbilical VEC wereobtained.These antibodies were proved to belong to immunoglobulin sub-classes(IgG_2,IgG_3,IgG_2).EC1 was shown to bind strongly to the vascularendothelial cell membrane,but not to react with peripheral lymphocytes andmonocytes in microcytotoxicity test.We also explored the possible applica-tion of the monoclonal antibodies in VEC typing.

Objective To investigate the prognosis and survival situation of the patients with high risk refractory lymphoma treated by autologous hematopoietic stem cell transplantation .Methods The follow up data in 110 patients with high risk refractory lymphoma treated by autologous hematopoietic stem cell transplantation in our hospital were retrospectively analyzed .The survival and prognosis factors of the patients after autologous hematopoietic stem cell transplantation were analyzed by using the Kaplan-Meier survival analysis and Cox proportional hazard regression analysis .Results The median survival time was 39 .4 months in 110 cases ,the 3‐year overall survival rate (OS) and progression free survival rate (PFS) were 80 .9% and 76 .4% respectively .The pa‐tients achieved CR status before transplantation(P=0 .016) and consolidation therapy after transplantation (P=0 .006) were the favorable prognostic factors of the patients undergoing transplantation .The prognosis in the patients with high LDH values ,IPI score>2 and bone marrow infiltration and HBV infection were poor(P<0 .05) .Conclusion Autologous hematopoietic stem cell transplantation can improve the long‐term survival rate in the patients with high risk refractory lymphoma .

Objectives To evaluate the potential of antisense c myc gene therapy for acute promyelocytic leukemia by investigating the biological effects and molecular mechanisms in induction of differentiation of HL 60 cell line using recombinant antisense c myc adenovirus (Ad AS c myc). Methods Cultured HL 60 cells treated with Ad AS c myc or Ad LacZ with polybrene and protamine sulfate were analyzed by X gal staining, morphology, MTT, flow cytometric analysis, RT PCR, and immunocytochemical techniques in vitro . Results HL 60 cells could be transfected effectively by Ad LacZ+protamine sulfate (79.8%). The level of c myc transcription and the version could be strongly inhibited by Ad AS c myc in the transfected HL 60 cells. Ad AS c myc could strongly inhibit the cell growth in HL 60 cells (51%). Ad AS c myc could reduce the ratio of nuclear/cytoplasm, and increase the activity of peroxidase in HL 60 cells. Ad AS c myc could lead to the blocking of G 0/G 1 phase in HL 60 cells. Ad AS c myc could also increase the expression of c fos in HL 60 cells. Conclusion The expression of Ad AS c myc can inhibit the growth and induce differentiation of HL 60 cells in vitro . The biological effects of Ad AS c myc may be closely associated with the activity of peroxidase and c fos gene in HL 60 cells. Ad AS c myc is of clinical potential in gene thera py for acute promyelocytic leukemia.

Objective:To establish a feasible method for extracting polysaccharide from the discarded fibrous roots of Radix Panacis Quingueforlii afterpanaquilon had been extracted, and determine the polysaccharide content. Methods:Enzymolysis technique and alcohol was applied for decolorization, and phenol-sulfuric-acid method was used to determine the active polysaccharide content.The content of trace element and heavy mental was measured by element-analyzer and atomic fluorescence photometer respectively. Results: The yield of polysaccharide from the fibrous roots was close (about 11.7%) to the main root.And the content of heavy metal can match the national standard.Conclusion:It is valuable to extract the polysaccharide from the discarded fibrous root of Radix Panacis Quingueforlii.

Objective:To establish a feasible method for extracti ng polysaccharide from the discarded fibrous roots of Radix Panacis Quingueforli i afterpanaquilon had been extracted, and determine the polysaccharide content. Methods:Enzymolysis technique and alcohol was applied for decolorization, and ph enol-sulfuric-acid method was used to determine the active polysaccharide conte nt.The content of trace element and heavy mental was measured by element-analyze r and atomic fluorescence photometer respectively. Results: The yield of polysac charide from the fibrous roots was close (about 11.7%) to the main root.And the content of heavy metal can match the national standard.Conclusion:I t is valuable to extract the polysaccharide from the discarded fibrous root of R adix Panacis Quingueforlii.

Objective To observe the efficacy of arsenic trioxide(ATO) combined all trans retinoic acid (ATRA) versus cytara‐bine (Ara‐C) combined ATRA in the treatment of acute promyelocytic leukemia(APL) .Methods We enrolled 65 patients in our department during the period between January 2002 and August 2008 ,and they were randomly assigned to receive ATRA combined ATO (treatment group ,n= 27) or ATRA combined DA ,HA ,NA which were major of Ara‐C (control group ,n= 38) .Then observe the differences of between the two groups ,such as complete remission(CR) ,the time to complete remission ,overall survival(OS) ,e‐vent free survival(EFS) ,the 5 years disease free survival (DFS) and adverse reactions .Results The CR rate of treatment group (ATRA + ATO) and control group (chemotherapy + ATRA) was 81 .48% and 68 .42% ,respectively ,and the time to complete re‐mission was (28 .50 ± 3 .97)d and (30 .56 ± 2 .39)d ,respectively ,showed that there was no statistical difference between the two groups ( P > 0 .05 ) .The 5 years DFS of the CR patients in the two groups was 51 .9% (ATRA + ATO ) and 50 .0%(Chemotherapy + ATRA) ,respectively ,showed that there was no statistical difference between the two groups(P > 0 .05) .The 5 years EFS of the CR patients in the two groups was 48 .1% and 39 .5% ,respectively ,showed that there was no statistical difference between the two groups(P> 0 .05) .The 5 years DFS of the patients in the two groups was 55 .6% and 67 .6% ,respectively ,showed that there was no statistical difference between the two groups(P > 0 .05) .Bone marrow suppression in the treatment group was significantly lower than in the control group(P< 0 .05) .Conclusion ATRA + ATO can prolong the CR rate ,OS ,EFS and 5 years EFS of newly diagnosed APL patients .ATRA combined with chemotherapy has similar efficacy ,ATRA + ATO has lower bone marrow suppression than the ATRA combined with chemotherapy ,thus may reduce the risk of early death .

Objective Gene microarray technology was used to investigate the differential gene expression of apoptosis related genes in NB4 cells induced by arsenic trioxide. Methods The databases of Evntrez and Human IPI were searched with "apoptosis or apoptotic" as the key words, and 1384 apoptosis related genes were found after the redundant genes were eliminated by chromosomal localization. The probes of these genes were designed using OligoArray 2.0, and then analyzed by BLAST. All the probes were immobilized on the glass slide, which were used as oligonucleotide microarray. After NB4 cells were treated with 2umol/L As2O3 for 48h, the total RNA were extracted. cDNAs of control group and test group were fluorescently labeled with Cy3 and Cy5, respectively, by RT-PCR. The fluorescent samples were hybridized with an oligonucleotide microarray containing 1384 apoptosis related genes to search for the differentially expressed genes in the cells with or without As2O3 treatment. The hybridization signals were scanned by oligonucleotide microarray, and then the fluorescent intensity of Cy3 and Cy5 and the ratio of two fluoresceins were analyzed using certain software. Then the differential expressed genes were analyzed after As2O3 treatment, in which the most distinctly differential expressed genes were chosen as targets, and through PCR amplification and gel electrophoresis the above genes were verified. Results There are 4 genes up-regulated and 12 genes down-regulated in expression in NB4 cells after 48h treatment with 2umol/L As2O3, which were in accordance with the results of RT-PCR and oligonucleotide microarray. Conclusion Differential gene expression in NB4 cells was induced by As2O3 treatment. These differentially expressed genes, with relation to signal transduction, transcription regulation, cell cycles, oxidation response, protein translation and cell differentiation, may play an important role in NB4 cell apoptosis.

Objective To understand the roles of JAK STAT pathway in the process of differentiation of human cord blood CD34 + hematopoietic stem cells into dendritic cells (DCs) by detecting the expressions and activation of JAK2 and STAT5. Methods CD34 + hematopoietic stem cells isolated from human umbilical cord blood and cultured for two weeks were induced to differentiate into DCs in vitro . Total cellular JAK2 and STAT5 and tyrosine phosphorylated protein stimulated by granulocyte/macrophage colony stimulating factor (GM CSF) at different time points (0, 7, and 14) during DC differentiation were detected by Western blotting. Results The amount of JAK2 protein was similar at 0, 7, and 14 d without GM CSF stimulation. With the differentiation of cells into DCs, the amount of tyrosine phospho JAK2 induced by GM CSF increased markedly. Both total cellular and tyrosine phospho STAT5 expression increased markedly during DC differentiation. Maximal tyrosine phospho STAT5 expression was later than JAK2. Conclusion JAK STAT pathway may take part in the signal mechanism of DCs differentiation from CD34 + hematopoietic stem cells stimulated by GM CSF.

Objective To establish the cell models of c-myb+/+ and c-myb+/-ES in vitro by gene targeting with ES cell culture system, with the aim to examine the detailed role of the transcription factor c-myb in haemopoietic commitment and differentiation. Methods Haemopoietic differentiation was initiated by seeding ES cells in LIF-free methylcellulose medium. The embryoids of c-myb+/+ and c-myb+/-ES were analyzed by methylcellulose colony assay and real-time PCR, the formative process of embryoids and the expression of relative genes were compared in each haemopoietic system at different differentiation stages and procedures. Results The formative process of embryoids was similar for both c-myb+/+ and c-myb+/-ES cells, but the size and frequency of EBs were reduced in the case of the c-myb+/-cells. Similar kinetics gain existed for the formation of CFU-Es in both groups, but the number in c-myb+/-group was less than that of c-myb+/+ group, and the colonies were generally smaller. BFU-E was first detectable on day 7, and the peak value emerged on day 10 in both groups. Similar kinetics gain existed for the formation of CFU-M in the two groups, but the number was larger in c-myb+/-group than that in c-myb+/+ group, while the number of CFU-GM in c-myb+/-group was less than that in c-myb+/+ group. Real-time PCR analysis showed no changes on the gene expressions of ?-globin, ?-globin, GATA3, CD34, Lys, c-fms and c-mpl in both groups. Conclusions Sub-level expressions of c-myb (55%) and c-myb+/-are sufficient to allow progenitor to expand, but they throw a negative influence on the terminal differentiation. Sub-level expression of c-myb may influence the erythropoiesis and granulocytic development, but throw no influence on the precursor cells to differentiate into macrophages. The expressive levels of c-myb have no effect on the expressions of ?-globin, ?-globin, GATA3, CD34, Lys, c-fms and c-mp.

Objective To investigate the relationship between genetic polymorphism of methylenetetrahydrofolate reductase (MTHFR) and the risk of acute lymphocytic leukemia (ALL). Methods Eighty-three patients with ALL and a cohort of 83 matched healthy objects were included, and DNA was extracted from their peripheral blood. PCR-RFLP was used to determine the genotypes of MTHFR C677T and A1298C. The adjusted odds ratio (OR) and 95% confidence interval (CI) were calculated using unconditional logistic regression model. Results It was found that the frequency of the MTHFR C677T TT genotype among patients was significantly different from that among control objects (P=0.008). The MTHFR C677T TT genotype had an increased risk of ALL compared with that of 677CC genotype (OR=3.229, 95%CI: 1.328-7.847, P=0.01). No significant association between the MTHFR C677T CT genotype or A1298C polymorphism and the risk of leukemia. Conclusion The present findings suggest that 677C→T polymorphism in MTHFR may be a genetic susceptibility factor for acute lymphocytic leukemia.