Objective To construet a novel DNA vaccine pcDNA3.1+/flk-1_((n1-4)) against tumor angiogenesis.Methods flk-1_((n1-4)) cDNA amplified by RT-PCR was inserted into eucaryotic expression vector pcDNA3.1+ and the recombinant plasmids pcDNA3.1+/flk-1_((n14)) was constructed.Confirmed by DNA sequence analysis,the recombinant plasmids was transfected into eucaryotic expression system COS-7 cells by lipofectamine.The protein expression of flk-1_((n1-4)) was detected by Western blot. Results Extracellular loop 1-4 of flk-1 was cloned and the recombinant plasmid was successfully constructed,which has been proved by DNA sequencing and compared with the data in Gene Bank.The protein expression of flk-1_((n1-4)) was detected in eucaryotic expression system COS-7 cells.Conclusions The DNA vaccine against tumor angiogenesis was successfully constructed,which found a basis for the further study on animal experiment and clinical application.

Objective:To investigate the strategies and methods of pancreatic cancer immunotherapy adopt cytotoxic T lymphocytes activated by dendritic cells(DCs),which modified by heat shock protein 70 peptide complex(HSP70-PC).Methods:HSP70 peptide complex from lumps of tumor-bearing mouse innoculated with pancreas cancer cell(MPC83)were purified by ConA-Sepharose and ADP-Agarose affinity chromatography.Dendritic cells derived from normal murine bone marrow were induced by GM-CSF and IL-4.MTT method was applied to analyses the proliferation activities of DCs.To achieve the specific T lymphocyte cells,DCs modified with HSP70 peptide complex were coculture with murine spleen lymphocytes for 48 hours.Then these activated lymphocytes were cocultured with MPC83 at ratio of effect-target 10∶1 and 20∶1 in vitro and the killing activities were detected by MTT method.Results:High purify HSP70-PC was obtained;104 DCs need 50～100ng of HSP70-PC to modify,100?g of HSP70-PC peptide complex can be extracted from per 1g lump;T cells stimulated by HSP70-PC modify DCs showed more specific cytotoxic activity to MPC83 than lymphocytes activated by DCs without HSP70-PC in vitro.Conclusion:We can extract high purity HSP70-PC for clinical individual immunotherapy from pancreatic cancer lump by using hypobaric affinity chromatography;DCs vaccine modified with HSP70-PC has significant kill effect against pancreatic cancer cell in vitro.

Objective To establish the quality standard of Tiaojing Zhixue Granules.Methods Radix Astragali,Radix Paeononiae Alba,Herba Ecliptae and Fructus Psoraleae in Tiaojing Zhixue Granules were identified by TLC;Paeoniflorin content in Tiaojing Zhixue Granules was determined by HPLC.Results The relevant spots in Radix Astragali,Radix Paenoniae Alba,Herba Ecliptae and Fructus Psoraleae can be identified by TLC.The content of paeoniflorin in Radix Paenoniae Alba can be determined by HPLC.The linearity of paeoniflorin was good in the range of 0.0968 ~ 0.4838 ?g(r = 0.9999).The average recovery of paeoniflorin was 99.71 % with RSD = 1.33 %.Conclusion The established quality standard is simple,feasible and repeatable,and can be used for quality supervisory of Tiaojing Zhixue Granules.

OBJECTIVE To investigate the risk factors and to take some useful measures to prevent and reduce infection in order to enhance medical quality,to ensure medical security,to strengthen hospital infection manangement and to prevent hospital infection effectively. METHODS We investigated the prevalence rate of hospital infection among our hospitalized patients in 2001,2003 and 2005, respectively. RESULTS The hospital infection rate was 4.6-6.42% in these years.Risk factors and the abuse of antibiotic were decreasing. CONCLUSIONS In order to control hospital infection rate,mensures should be taken including intensively monitoring the departments with high infection rate,strengthening hospital operation,rationally using the antibiotics,and studying the management for hospital infection.

Gastrointestinal bleeding is a common complication of gastrointestinal diseases and gastrointestinal surgery, which may lead to hemorrhagic shock or cause death if not treated properly and promptly.Currently, endoscopic treatment for gastrointestinal bleeding includes clip closure, endoscopical injection of saline/adrenaline, argon plasma coagulation (APC), electrocoagulation and heater probe coagulation etc..Endoscopic clip closure mainly includes traditional closure through-the-scope clip (TTSC) and more novel closure over-the-scope clip (OTSC).This article reviewed the use of OTSC in the treatment of gastrointestinal bleeding.

Objective To study the immunophenotypic feature of CD+2 adult B cell acute lymphoblastic leukemia (CD+2 B-ALL) and provide evidences for the diagnosis, therapy and prognosis. Methods The immunophenotypes of 18 cases of adult CD+2 B-ALL and 68 cases of adult CD-2 B cell acute lymphoblastic leukemia(CD-2 B-ALL) were assayed by a panel of monoclonal antibodies (McAbs) with FACS. Results The age of CD+2 B-ALL adults was younger compared to that of CD-2 B-ALL. The patients in the CD; group were similar to CD-2 group for the expression of most of the cell surface antigens assayed. However, it was notable that expression frequencies of CD10 [(73.78±26.67) %] in CD+2 B-ALL was higher than those in CD; B-ALL [(52.84±35.25) %] (t = 2.35, P<0.05) and CD33 [(15.46±27.41) %] were significantly lower than that of CD2 BALL [(31.15+27.72) %] (t = 2.16, P<0.05). Both groups showed high positive expression for CD34 72.2 % (13/ 18) and 80.9 % (55/68), respectively (χ2 = 0.64, P >0.05). The CD20 expression in CD+2 B-ALL was lower than that in CD-2 B-ALL significantly (χ2 = 11.38, P <0.05). The myeloid antigen (CD13 or CD33) expression in CD+2 B-ALL 44.4 % (8/18) was lower than that in CD-2 B-ALL 72.1 %(49/68) (χ2 = 4.86, P <0.05). Conclusion Patients in the CD+2 B-ALL were similar to CD-2 B-ALL for expression of most of the cell surface antigens assayed. CD+2 B-ALL showed low expression frequency of myeloid antigens (CD13, CD33) and CD20 antigens than CD-2 B-ALL. These results suggested that those with the CD+2 B-ALL immunophenotype generally presented with favorable prognostic features and usually could achieve good outcomes after treatment.

Objective To assess the clinical value of 131I-metaiodobenzylguanidine (MIBG) scintigraphy in pheochromocytoma. Methods A total of 430 patients with clinically suspected pheochromocytoma underwent 131I-MIBG whole body scintigraphy, 326 among them underwent B-ultrasound, 400 for CT and 77 for MR examination respectively. While 178 among them were diagnosed with pathology and the others were diagnosed clinically. Results Of all the patients, 108 were diagnosed pheochromocytoma, including 89 131I-MIBG scan positive and 19 negative. The sensitivity, specificity and accuracy of 131I-MIBG were 82.41%, 100% and 95.70%, respectively. 131I-MIBG scan detected 90.00% of unilateral adrenal, 45.45% of bilateral adrenal, 85.71% of ectopic and 66.67% of malignant lesions, respectively. The proportion of patients with positive 131I-MIBG scan increased from 20.69% in all patients to 35.15% in patients with clinical symptoms and positive conventional imaging (at least one of B-ultrasonography, CT or MR was positive) and 64.58% in those with clinical symptoms, positive conventional imaging, and elevated 24 h urinary catacholamines. In 59 patients with adrenal incidentaloma, 8 were scan-positive and all had confirmed pheochromocytoma, while 2 of scan-negative patients also had confirmed pheochromocytoma. Conclusion 131I-MIBG scintigraphy is the first choice for the diagnosis of both adrenal and extra-adrenal pheochromocytoma. However, it is inappropriate to take this method as the initial screening approach.

Human tissue kallikrein-binding protein (Kallistatin, KAL), a secretory protein that participates in the regulation of multiple signaling pathways by binding to the extracellular receptor, however, at present has not been reported about the intracellular activity, and whether it has the similar biological activity with extracellular activity. Here we constructed no signal peptide KAL (NSK) into the adeno-associated virus vector to explore the intracellular activity of KAL. Both the endothelial cell and lung cancer cells could express KAL, but not secreted after rAAV2-NSK transfection. The proliferation and migration of human umbilical vein endothelial cells (HUVECs) were inhibited, but the apoptosis rate was not affected. The proliferation rates, mobility and tubule formation of all the three tested lung cancer cells, such as NCI-H446, NCI-H460 and A549, were inhibited to different extents. This cellular study not only confirmed the intracellular activity, but also suggested it may serve as a kind of "balance factor" in multi-targeted controlling, which may provide a new train of thoughts to explain the regulatory contradiction in PI3K-Akt signaling pathways by KAL.

Objective To investigate the clinical and laboratory features of acute promyelocytic leukemia (APL).Methotis 513 APL patients in the last two decades were retrospectively analyzed in this research.We investigated the clinical features including age,sex,abnormality of peripheral hemogram before treatment.therapeutic effect and follow-up and laboratory data such as morphology,immunology,cytogenetics and molecular biology(MICM).Results The median age of the APL patients was 33 years old and the ratio of male and female was 1.21:1.Before treatment,the median level of WBC was 4.3×109/L and the deteetion rate of abnormal promyelocyte on blood film was 85.8%;with immunophenotypie detection,the expression levels of CD117、CD34、HLA-DR、CD7、CD14 and CD19 in APL were found to be lower and the expression 1evels of CD2、CD33 and MPO higher than those in other subtypes of acute myelocytie leukemia(AML)(beth P<0.01).Specific abnormal chromosome t(15;17)was detected in 91.7%of the patients,of whom 75.9%had standard translocation of t(15;17),being the most common one and 15.8% of the patients had t(15;17)with additional abnormal chromosome.There was only 7.5%of the patients with nolnlal karyotype.However,the presence of both simple translocation and complex translocation was seldom seen.With molecular biological detection.PML/RARα fusion gene positive rate was 99.6%.In a relativelv long clinical follow-up,we found that the complete remission(CR)rate in APL patients was 84.7%.incidence of DIC was 13.4%and five-year survival rate was 30.7%.111e median count of WBC in CR group was lower than that non-remission group(P<0.01).There were no significant differences on expressions of CD34 and CD2 and changes of cytogenetics between the two groups(P>0.05).Conclusions Comprehensive evaluation of MICM could be of important significance in the diagnosis and prognosis iudgrnent for APL patients.The CR rate in these patients with high WBC eount was considerable low.

Objective To study the changes and significance of CD4+CD25+ regulatory T cells and C-reactive protein(CRP) in patients with acute exacerbation of chronic obstructive pulmonary disease(COPD). Methods Flow cytometry was used to detect the frequency of CD4+CD25+ regulatory T cells in peripheral blood from 36 patients with acute exacerbation of COPD( COPD group) and 36 patients with clinical stability of COPD(control group one)and 36 normal individuals(control group two). The level of CRP was detected routinely. Results The ratio of CD4+CD25+ regulatory T cells number in peripheral blood of COPD group to the total number of CD4+T cell was (2.56±1.83 )%, and it was significantly decreased compared to the other two groups (P all<0.01 ). The level of CRP in COPD group was markedly higher than that in the other two groups (P all<0.01 ). The level of CD4+CD25+ regulatory T cells in patients with acute exacerbation of COPD had negative relation with CRP. Conclusions CD4+CD25+ regulatory T cells participate in inflammatory response. The proportion of CD4+CD25+ regulatory T cells decreases in patients with acute exacerbation of COPD, and it may result in maladjustment of cytoimmunity.