Neutrophils play important role in anti - tumor response of tumor-bearing host as a kind of important effector cells. In order to identify the anti-tumor mechanisms of G-CSF gene therapy, we investigated the number and functions of the peripheral neutrophils in the C-26 colon adenocarcinoma-bearing mice receiving G - CSF gene therapy and high - dose chemotherapy. After G - CSF gene therapy, the number of neutrophils in peripheral blood was increased markedly in C-26 mice receiving high - dose 5 - Fu as compared with control groups including in vivo administration of rhG - CSF. The more potent cytotoxicity to C - 26 cells could be detected. The phagocytic activity, the secretion of IL-1, TNF, NO of the neutrophils were significantly enhanced. These data showed that G - CSF gene therapy can increase the number of neutrophils, activate the functions more effectively than in vivo administration of rhG - CSF.

Granulocyte colony - stimulating factor (G - CSF) is a hematopoietic growth factor that is responsible for the differentiation and proliferation of hematopoietic progenitor cells to mature granulocyte, and can increase the number of peripheral neutrophils. It has been demonstrated that it could inhibit the metastasis of the murine tumors in spontaneous and experimental metastasis models by in vivo administration of recombinant human G - CSF. In order to examine the antitumor effect of G - CSF gene therapy on mice receiving high - dose chemotherapy, C - 26 colon adenocarcinoma - bearing mice which were prepared by S. c. injection of 1?105C-26 cells were i. p. injected with rhG-CSF (2?g/day?14day) or implanted with 1?107 collagen encapsulated NIH3T3-G-CSF cells which secrete high level of G - CSF after gene transfection. In our experiment, rhG-CSF could inhibit the tumor growth and extend the survival time of early stage C-26 bearing mice. However, G - CSF gene therapy could inhibit the tumor growth and prolong the survival both in early or middle stage C-26 mice. The results showed that both rhG - CSF and G - CSF gene therapy have exact antitumor effect and G - CSF gene therapy show more effective than rhG - CSF in vivo. Then we investigated the therapeutic effects of G - CSF gene therapy on C-26-bearing mice receiving high - dose chemotherapy (5-Fu 150mg/mice i. p.) . More effective results could be observed in C-26 - bearing mice receiving high dose chemotherapy after G - CSF gene therapy. The results also suggested that G-CSF gene therapy can inhibit the tumor growth more effectively both in C-26-bearing mice or C-26-bearing mice receiving high - dose chemotherapy.

To observe the effect of IL-2 gene modification on the character of biology and function in dendritic cells( DC) and to investigate the immune mechanism of specific anti-tumor of IL-2 gene modification in DC. Methods: DCs were prepared from mouse bone marrow and genetically modified by IL-2 adenovirus. Then observe the changes of DC morphology by scanning electro-microscopy, analyzed molecules on DC by FACS, examined the expression of IFN-? mRNA in DC by RT-PCR.The stimulatory capacity of DC to T cells detected by MLR their capacity of antigen present were measured by 3H-TdR mix into assay. Results: After IL-2 gene modification, the morphology of DC was changed, its pseudopod was more and longer. The expression of Ia, B7-1, B7-2 and CD40 molecules was more on DC surface. The IFN-7 mRNA was expressed in the DC-EL-2 and DC-rhIL-2.DC-IL-2 could stimulate allogeneic T cells more potently and IL-2 gene-modified DC could induce more potent antigen-specific autogeneic CTL. Cooclusioo: IL-2 genetic modification can promote DC growth and up regulate their expression of membrane immune molecules that are relevant for antigen presentation of DC,and enhanced the biologic activity of the DC.

Objective:To evaluate the effects of dibutyl phthalate on rat sperm production and quality. Methods:Totally 150 male rats were randomly divided into the low dose group (50 mg·kg-1), the middle dose group (200 mg·kg-1), the high dose group (1 000 mg·kg-1 ) , the blank control group and the solvent control group ( peanut oil as the control) with 30 ones in each. After continu-ous administration for every 30 days, 10 rats from each group were anatomized, the weight of testes and epididymides were determined, and one side of epididymis was used to carry out the sperm analysis including counting, survival rate and morphology. Results:After intragastric administration for 90 days, the sperm count and survival rate, the weight of testis and epididymis and organ coefficient in the middle dose group and high dose group were decreased significantly(P<0. 05 or 0. 01). Conclusion:The long-term administration of dibutyl phthalate at high dose exhibits notable toxicity on rat reproductive function.

Objective:To investigate the specific immune act ivated effects in the mouse immunized by MHC I-restriced tumor antigen peptide pulsed dendritic cells(DC) with IL-2 gene modified.Methods:DC were transfected with IL-2 gene via adenovirus vector(DC-IL-2) the mRNA of mIL-2 was examined by RT-PCR.Syngeneic mice were immunized by DC-IL-2 pul sed with MHC-I-restricted Mutl tumor antigen peptiede of 3LL Lewis lung carcin oma (DC-IL-2-Mutl),the effects of lymphocyte subsets in the draining lymphoid node from the tumor antigen pulsed DCs with gene modified immunized mice by flo w-cytometric analysis(FACS) analysis.Results:The mRNA of mIL-2 was expressed in the DCs.The ratio of CD8+T cells was hosted in the dra ining lymphoid node from Mutl pulsed DCs(DC-Mutl) immunized mice.The ratio of C D8+T cells and NK cells were all hosted obviously in the draining lymphoid nod e from DC-IL-2-Mutl immunized mice .Conclusion:Immunized mice with DC-IL-2-Mutl can induce the mouse specific antitumor immunized affect in vivo and can activate a variety of immunized effect in the mouse.

Objective To explore the regulating effects of Egr-1 promoter activated by ionizing radiation (IR) and doxorubicin (ADM) on the expression of granulocyte-macrophage colony-stimulating factor (GM-CSF) genes. Methods The human GM-CSF cDNA and enhanced green fluorescent protein (EGFP) cDNA were linked together with IRES(internal ribosome entry site) and then inserted into the expression vector pCIneo under control of the Egr-1 promoter(Egr-EG). The vector was transferred into human bone marrow stromal cell line HFCL by liposome transfection. And the cells were exposure to ADM and IR. The activity of EGFP in HFCL/EG cells were detected by FACS. The effect of N-acetylcysteine on the expression of EGFP following exposure to ADM and IR was examined. The amounts of GM-CSF in HFCL/EG after chemotherapy or radiation were measured with ELISA. The effects of GM-CSF in HFCL/EG cultural supernatants on expansion of CFU-GM derived from cord blood were also studied. RT-PCR analysis for the expression of GM-CSF mRNA in HFCL/EG after exposure to ADM or IR. Results The percentage of EGFP+ HFCL/EG cells exposed to ADM and IR was increased compared with non-treatment group (1.2 % and 15.2 % vs 18.2 %, t = 5.11, P < 0.01). The levels of secreted GM-CSF in HFCL/EG cells exposed to ADM and IR was increased (P < 0.01), but no difference between ADM group and IR group (P 0.05). The expression of EGFP by HFCL/EG treated with ADM and IR was significantly decreased by N-acetylcysteine. The effects of GM-CSF in HFCL/EG cultural supernatants on expansion of CFU-GM in ADM group and IR group were significantly higher than that in HFCL group and non-treatment group. However, The CFU-GM count of IR group was higher than that of ADM group. The expression of GM-CSF mRNA in HFCL/EG cells exposed to ADM and IR was significantly increased(t = 4.37, P < 0.01). Conclusions GM-CSF gene expression regulated by Egr-1 promoter induced by ADM and IR could help the recovery from hematopoietic injury.

Objective: To determine the anti rejection effects of CTLA 4 Ig fusion protein on cardiac allografts in mice and to discuss its mechanism in vivo . Methods: BALB/c recipients were performed cervical heterotopic heart transplantation to receive C57BL/6 donor hearts with a cuff technique. BALB/c recipients were intraperitoneally injected with CTLA 4 Ig [100 ?g/d?15 times], control immunoglobins and PBS to observe the survival time of allografts with ECG. The hyporesponsiveness of splenic T cell, the polarization of the T subsets were analyzed after the recipients treated with CTLA 4 Ig. Results: After treated with CTLA 4 Ig, the survival of cardiac grafts was significantly prolonged compared with the control groups, and more than 40% cardiac grafts survived over 2 months. The splenic T cells isolated from recipients did not respond to restimulation of donor splenocytes in MLR, but did exhibit the capacity to proliferate in response to C3H splenocytes(third party).The levels of IL 2 and IFN ? decreased and the level of IL 10 increased in CTLA 4 Ig treated mice. Conclusion: Administration of CTLA 4 Ig can induce donor specific tolerance, which induce T subsets to polarize toward Th2 subset and hyporesponsiveness to alloantigen, and prolong the survival time of the cardiac grafts effectively. [

目的探索脑瘫患儿早期发现、早期干预的工作模式。方法临床、保健、康复相结合 ,采用产前检查、孕妇学校、产时儿科医生进产房、新生儿行为神经评定测定、高危儿抚触训练室、婴儿期儿童保健监测、各街道防治站常规体检筛查转诊等多种形式结合早期发现并干预脑瘫高危儿。结果此工作模式较模式前显著提高了脑瘫的早期诊断率及康复疗效。结论临床、保健、康复相结合的方式整合了医疗资源,起到了促进脑瘫早发现、早诊断、早治疗的作用 ,值得推广。

OBJECTIVETo investigate the effects of IL-2 gene modification enhancement of the antigen-presenting function of the mouse bone marrow derived dendritic cells and on the activation of CTL induced by MHC class I molecule restricted antigen peptides as well as the related immunological mechanisms.

METHODSDCs were prepared from mouse bone marrow and modified by recombinant IL-2 adenovirus (DC-IL-2). The IL-12 and IFN-gamma levels in culture supernatant of DC and CTL were examined by ELISA, the expression of costimulatory molecules and fluorescent intensity of endocytosis of OVA-FITC in DC by FACS, the capacity of presenting 3LL cell tumor antigen by (3)H-TdR incorporation method, the MHC class I-restricted tumor-antigen-peptide Mut1 of 3LL cells pulsed DC-IL-2 to induce CTL cytotoxicity by (51)Cr 4-hr releasing assay.

RESULTSAfter IL-2 gene modification, DC-IL-2 could produce high level of IL-12 [(78.4 +/- 6.6) pg.(1 x 10(6) cells)(-1).ml(-1)]. The expression of costimulatory molecules on DC-IL-2 was increased, the fluorescent intensity of DC captured OVA-FITC was enhanced, and the proliferation of allo-T cells from 3LL bearing mouse pulsed with Mut1 was also enhanced. Mut1 antigen peptide pulsed DC-IL-2 could induce more potent antigen-specific CTL cytotoxicity and excrete high concentration of IFN-gamma [(1 168 +/- 58.4) pg/ml] in vivo.

CONCLUSIONIL-2 gene modification of DC can activate second signal for DC presenting antigen, and enhance the function for capturing and presenting tumor antigen. DC-IL-2 pulsed with MHC class I restricted tumor-antigen-peptide can induce specific anti-tumor immune response more effectively. Owing to IL-2 gene modification, the functions of IL-12 excretion and T cell activation of DC were promoted, so that the capacity of CTL excreting IFN-gamma was enhanced, which are relevant to the immune mechanism.

Adenoviridae ; genetics ; Animals ; Antigen Presentation ; immunology ; B7-1 Antigen ; genetics ; metabolism ; Dendritic Cells ; cytology ; immunology ; Female ; Interleukin-12 ; secretion ; Interleukin-2 ; genetics ; Lymphocyte Activation ; genetics ; immunology ; Mice ; Mice, Inbred C57BL ; Recombination, Genetic ; genetics ; immunology ; T-Lymphocytes, Cytotoxic ; cytology ; immunology

The transcriptional repressor RE1 silencer transcription factor(NRSF/REST) is an important factor that restricts some neuronal traits in neurons.Since these traits are also present in pancreatic islet cells,NRSF-regulated genes involved in islet function are searched.A NRSE-like motif was analysed in human insulin promoter.The role of NRSE was evaluated by generating a model of insulin-secreting cells that firmly express NRSF.The presence of NRSF led to a decrease in activity of human insulin promoter by stable or transient transfection with human insulin-promoter luciferase.The predicted NRSE-like motif also confers NRSF-dependent transcriptional repression in the context of a surrogate gene promoter.Specific binding activity of NRSF/REST to the NRSE-like motif was confirmed by EMSA.Moreover,the binding activity is competed by consensus NRSE sequence.These data showed that human insulin promoter is regulated by the transcriptional repressor NRSF/REST via the NRSE-like motif.