Objective To study the survivin expression in mononuclear cells of both bone marrow and peripheral blood in childhood acute leukemia(AL).Methods Twenty-one children with AL were divided into newly diagnosed group(n=12) and complete remission(CR) group(n=9).The control group was consisted of 15 children without tumor.Mononuclear cells were separated from bone marrow and peripheral blood (PBL) by gradient density centrifugation and RNA was extracted,and RT-PCR was performed to detect survivin mRNA.Results In newly diagnosed AL group,6 out of 12 patients had positive survivin expression in bone marrow(50%) and 4 out of 11 patients in the PBL(36.37%),both of which were higher than the control group(P0.05).In 9 CR patients,all the patients had the negative expression in the PBL,but 1 positive expression in bone marrow(11.11%).The positive expression in CR group was significantly lower than the newly diagnosed AL group.The only one patient with positive survivin expression in CR group had relapse 7 weeks later.In newly diagnosed patients,the average immature leukocytes were(76.43?43.28)?10~(9)/L in the patients with positive survivin expression and(13.85?5.86)?10~(9)/L in those with negative survivin expression(P

Glycogen synthase kinase 3β (GSK3 β) is a conserved cytoplasmic serine/threonine protein kinase.It can not only negatively regulate the classical Wnt and Akt signal pathways which are associated with leukemia,but also positively regulate the NF-κB,non-classical Wnt and lysosomal apoptosis signal pathways,thereby regulating the proliferation,differentiation and apoptosis of leukemia cells.At present,a variety of GSK.3 β inhibitors and activators have been reported in the researches of leukemia,which play important roles in reducing the adverse reactions of chemotherapy,reversing multidrug resistance and enhancing killing effect to leukemia cells.GSK3β is expected to become a target of leukemia molecular targeted therapy.

Objective To study the effect of survivin gene expression on the lymphocyte’s proliferation and function in cultured K562 cells. Methods The constructed recombinant vector pEGFP-C1-survivin and the plasmid pTZU6+1-survivin encoding short hairpin RNA of survivin were transfected into K562 cells respectively to generate K562/survivin+ cells and K562/survivin-cells. K562/survivin+ cells were selected by G418. The survivin mRNA and protein levels in the 3 kinds of cells (K562/survivin+,K562 and K562/survivin-) were detected by semi-quantitative RT-PCR and immunohistochemistry methods. The peripheral blood mononuclear cells (PBMC) from healthy subjects were co-cultured with these 3 cells respectively in mixed lymphocyte-tumor cell culture (MLTC). Lymphocyte proliferation in the supernatant was evaluated by MTT assay. Nature killer activity was detected by flow cytometry and IFN-? level was measured by ELISA assay. Results Proliferation index in K562/survivin+ group was much lower than that in the other 2 groups (P

Objective To construct an recombinant adenovirus vector of vascular endothelial growth factor small interfering RNA(Ad5-VEGFsi) and to observe its inhibitory effect on the cell proliferation of human leukemia K562 cells. Methods The specific human VEGF siRNA was subcloned into the shuttle plasmid pSES-HUS and then cotransfected with plasmid pAdeasy-1 to produce pAd5-VEGFsi by homologous recombination. The identified recombinant plasmid pAd5-VEGFsi was packaged and amplified in 293 cells. At 72 h after the transfection of Ad5-VEGFsi into K562 cells, VEGF mRNA expression and the protein level of VEGF in the cell culture supernatant were determined by RT-PCR and ELISA, respectively. Cell proliferation was measured by MTT assay and cell apoptosis by flow cytometry. Results The recombinant adenovirus Ad5-VEGFsi was successfully constructed. Ad5-VEGFsi with the titer of 4.6?1011 pfu/ml was harvested by CsCl gradient purification. Compared with those in the control, EGF mRNA in cells decreased by 66.55% (P

ObjectiveTo explore the inflammatory factors C-reactive protein (CRP),interleukin (IL)-6,tumor necrosis factor (TNF)-α and white blood cell (WBC) count differences between acute myocardial infarction(AMI ) thrombolysis treatment and unthormbolysis treatment,and find out the relevance between the inflammatory factors and the prognosis.MethodsAccording to the condition of accepting AMI thrombolysis treatment,the 229 patients of AMI were divided into the thrombolysis group( 131 cases) and the unthrombolysis group(98 cases).The levels of myocardial troponin I (cTnI),creatine kinase(CK),creatine kinase isoenzyme-MB (CK-MB) were detected at the time of patients sent into the hospital for the immediate,6-hour later and 24-hour later.After 6-month's follow-up,prognosis was compared between two groups.ResultsTwenty-seven cases lost in the thrombolysis group.One case died within 6 months and the mortality was 1.0％(1/104) in the thrombolysis group,and 6 cases died within 6 months and the mortality was 6.1％(6/98 ) in the unthrombolysis group.There was significant difference between two groups (P ＜ 0.05 ).The levels of CK,CK-MB in the thrombolysis group advanced,and compared with that in the unthrombolysis group,there were significant differences (P ＜ 0.05 ).The levels of TNF-α,IL-6 in the thrombolysis group were significantly higher than those in the unthrombolysis group (P＜ 0.05),and CRP and WBC count had no significant differences between two groups (P ＞ 0.05).The repatency rate was 79.4％( 104/131 ) in the thrombolysis group,the levels of TNF- α,IL-6 in repatency patients were higher than those in non-repatency patients.There were significant differences(P ＜ 0.05 ).ConclusionsThe thrombolysis is an effective way to cure AMI.The increase of TNF- α,IL-6 after the thrombolysis is considered to be related to reperfusion injury,and CRP,IL-6,TNF-α and WBC count can forecast the inflammation of myocardial necrosis and take an impotant role in predicting the prognosis of the AMI.The antiinflammatory and antioxidation treatment is significant to improve the prognosis of the AMI.

Objective To investigate the expression of Foxo3a in the lumbar dorsal root ganglia (DRG) after sciatic nerve injury in rats. Methods Forty-two adult rats were randomly divided into control group and experimental group. Rats in experimental group were subjected to sciatic nerve clamp. Expression and distribution of Foxo3a, growth associated protein (CAP) and axon regeneration in DRC were detected by Western blot and immunofluorescent staining. Results (1) After sciatic nerve injury, Foxo3a protein levels began to reduce at day 1, reached the lowest level at day 2 and then gradually rose to nomal level at four weeks after sciatic nerve injury. Proliferating cell nuclear antigen (PCNA) and GAP-43 were up-regulated from day 2, and the level of PCNA reached peak at day 7 after sciatic nerve injury in DRC. (2) Down-regulation of Foxo3a was observed predominantly in neurons and glial cells. PCNA mostly expressed in glial cells and hardly in neurons. Conclusion Down-regulation of Foxo3a in lumbar DRG may correlate with axonal regeneration and the proliferation of glial cells after sciatic nerve injury.

Objective To investigate the expression of Foxo3a and p27kip1 in lumbar dorsal root ganglia (DRG) after injury of sciatic nerve in rats. Methods Adult rats were randomly divided into control group and experimental group. The rats in experimental group were subjected to sciatic nerve clamp.Expression and distribution of Foxo3a and p27kip1 and cellular proliferation and axon regeneration in DRG was detected by western blot and immunohistochemistry. Results Foxo3a protein levels begined to reduce at 1 day (7.0 ± 3.5), reached valley at 2 day (6.0 ± 3.8) after injury, and following Foxo3a downregulation, p27kip1 protein levels begined to decrease at 2 day (29.0 ± 3.5), reached valley at 7 day (21.0± 3.0) after injury. Down-regulation of Foxo3a and p27kip1 was expressed predominantly in neurons and glial cells by double immunolabelling. Foxo3a and p27kip1 were expressed in neurons [(37.8 ± 5.7)%, (43.3 ±4.3)%] and glial [(22.4 ± 3.9)%, (13.8 ± 3.2)%] cells in sections of DRG at 2 day after injury less than neurons [(73.6 ± 2.5)%, (84.1 ± 3.7)%] and glial [(61.3 ± 4.4)%, (68.7 ± 5.6)%] cells in sections of normal DRG. Proliferating cell nuclear antigen (PCNA) and GAP-43 were up-regulation from 2 day, and PCNA reached peak at 7 day after injury.The glial cells were the main type of cellular proliferation.Conclusion Down-regulation of Foxo3a and p27kip1 in lumbar DRG is correlated with the proliferation of glial cells and axonal regeneration after sciatic nerve injury.

Objective To investigate the influence of very late antigen-4 (VLA-4) antibody and VLA-4 antibody combined with VP_(16) in vitro on childhood leukemic cells' apoptosis and explore the protection of bone marrow stromal cells (BMSC) upon leukemic cells and its related mechanisms. Methods Leukemic bone marrow stromal cells were isolated by human lymphocyte separation medium and in vitro culture of BMSC (adherent) and leukemia cells (suspended) BMSC+leukemic cells group were as control. Then VLA-4 antibody and/or VP_(16) were added respectively to VLA-4 antibody group, VP_(16) group and VLA-4 antibody combined with VP_(16) group to detect the apoptosis of leukemic cells in different groups through Annexin V-FITC double-labeled flow cytometry and the expression of Survivin, bcl-2 genes in each group of leukemic cells detected by RT-PCR. Results The results showed by flow cytometry that compared with the control groups, for 12 h or 24 h, the early and total apoptosis rates of leukemic cells of the three experimental groups were significantly increased(P <0.05); the early and total apoptosis rates of leukemic cells treated with VLA-4 antibody combined with VP_(16) group was markedly increased, compared with the control group (P <0.05); the comparison of the early and total apoptosis rates for the three experimental groups between 12 h and 24 h was significantly different (P<0.01). Moreover, RT-PCR results showed that the expression of Survivin and bcl-2 genes of leukemic cells in three experimental groups was reduced in varying degrees and the reduction of VLA-4 antibody combined with VP_(16) group was the most obvious. Conclusion BMSC plays a protective role on leukemic cells, and VLA-4 antibody can block the adhesion between BMSC and leukemic cells promoting leukemic cells apoptosis and enhance the sensibility of apoptosis of leukemic cells induced by chemotherapeutics.

Objective To investigate the expressions of Livin ? and Livin ? in marrow mononuclear cells(MMNCs)of childhood acute leukemia(CAL)and explore the clinical significance.Methods Real-time quantitative PCR and Western blot were used to detect the mRNA and protein expressions of Livin ? and Livin ? in MMNCs of CAL,respectively.Results Both Livin? and Livin? expression rates and expression levels were higher in preliminary diagnosis group(n=51)of CAL,including acute lymphoblastic leukemia(n=39)and acute myeloid leukemia(n=12),compared with the control group(P5%)to induction chemotherapy of ALL than in the patients without chemotherapy(P

Aim To investigate the cytotoxic activity of HDAC inhibitor Trichostatin A (TSA)combined with anticancer drugs targeting DNA on T24 bladder cancer cell line.Methods MTT assay was used to detect the inhibitory rate of TSA alone or combined with ADM, MMC and DDP respectively on T24 bladder cancer cell in cancer cell proliferation by administering TSA alone or combined with ADM, MMC and DDP respectively.Jin′s equation was used to evaluate the efficacy of drug combination. Results The growth inhibitory rate of TSA combined with ADM, MMC and DDP respectively was in a concentration-dependent manner. The synergism of TSA combined with MMC was most significant. When administered in lower or moderate concentration, TSA combined with ADM or DDP in lower or moderate concentration demonstrated synergic effect too.Conclusion HDAC inhibitor TSA enhances the cytotoxic activity of anticancer drugs targeting DNA on bladder cancer cells and is promising to be used in chemotherapeutic regimens for advanced bladder cancer.