胃泌素对大鼠胃粘膜环氧合酶及生长因子表达的影响

目的：探讨胃泌素对大鼠胃粘膜环氧合酶（COX）及生长因子表达的影响。方法：雄性SD大鼠皮下注射胃泌素1 μg/kg、10 μg/kg或100 μg/kg，Western blot和免疫组化检查胃粘膜COX-1、COX-2、肝细胞生长因子（HGF）和肝素结合表皮生长因子样生长因子（HB-EGF）表达。评价胃泌素受体拮抗剂YM022对COX-1、COX-2、HGF和HB-EGF表达的影响。结果：胃泌素剂量依赖性地增加大鼠胃粘膜COX-2和HB-EGF表达，而对COX-1和HGF表达无明显影响；YM022阻断胃泌素诱导的COX-2和HB-EGF表达。 结论： 胃泌素调节大鼠胃粘膜COX-2和HB-EGF蛋白表达，提示COX-2和HB-EGF参与与胃泌素相关联的胃粘膜增生和胃癌等的发病过程。     

血管内皮生长因子对外周血内皮祖细胞功能的影响

目的: 观察不同浓度血管内皮生长因子(VEGF）对体外培养人外周血内皮祖细胞（EPCs）生物学功能的影响，探讨VEGF促进EPCs分裂生长的合适浓度。方法: 密度梯度离心法获取人外周血单个核细胞（MNCs），接种至人纤维连接蛋白（HFN）包被的培养板上，培养4 d后收集贴壁细胞，换用配有不同浓度（对照组、10 μg/L、20 μg/L、50 μg/L）VEGF的培养基继续培养3 d后进行细胞免疫组化鉴定EPCs，采用MTT比色法、改良的Boyden小室和黏附能力测定实验，观察EPCs的增殖、迁移和黏附能力。结果: 从人外周血能成功分离EPCs细胞，并能分化为血管内皮细胞；从冠心病患者分离的EPCs增殖能力较非冠心病患者弱；VEGF在较低浓度（10 μg/L和20 μg/L）时即能显著促进EPCs生长的各项生物学指标，但高浓度（50 μg/L）时并不能进一步增强这一效应；低浓度（10 μg/L）下VEGF对冠心病患者作用较非冠心病患者弱，高浓度时对两者促进作用相近。结论: 冠心病患者EPCs功能显著减弱，较低浓度的VEGF即可显著增强EPCs的各项生物学功能，可能对损伤血管的再内皮化有益。     

急性髓系白血病血管内皮生长因子表达与预后的研究

目的 :观察人白血病细胞系血管内皮生长因子 (VEGF)表达水平 ,研究急性髓系白血病 (AML)患者血清VEGF表达水平与预后的关系。方法 :采用酶联免疫吸附法 (ELISA)对 4 9例初治、10例复发AML患者血清及人白血病细胞系U937、K5 6 2、HL - 6 0、TF - 1和NB4培养上清液 (4 8小时 )VEGF表达水平进行检测。结果 :五种人白血病细胞系培养上清液中均测到VEGF高表达。 4 9例初治、10例复发AML患者的血清VEGF表达水平分别为 2 0 1 17± 110 93pg ml和 2 32 5 9± 118 6 2pg ml,均明显高于正常对照组 (12 5 6 2± 4 5 4 3pg ml;p <0 0 5 )。初治AML患者中VEGF高表达组 (>2 0 1 17pg ml)完全缓解 (CR)率为 4 8% ,低表达组 (<2 0 1 17pg ml)CR率为 77% ,两者比较差异显著 (p<0 0 5 )。结论 :血管内皮生长因子在刺激白血病细胞增殖、迁移中发挥重要作用。AML患者血清VEGF水平与预后具相关性     

A synthetic 10-kD heat shock protein (hsp10) from Mycobacterium tuberculosis modulates adjuvant arthritis

The heat shock protein, hsp10, is an abundant protein in Mycobacterium tuberculosis (Mtb), its nucleotide sequence encoding a protein of 99 amino acids with a molecular mass of 10±7kD. This sequence is phylogenetically conserved, being represented by the GroES homologue of Escherichia coli. Hsp 10 and GroES are members of the chaperonin 10 family of molecular chaperones, and GroES is necessary for the optimal activity of GroEL, a member of the chaperonin 60 family and the E coli homologue of mycobacterial hsp65. Since hsp65 has been implicated in both experimental and human rheumatoid arthritis, we aimed to assess the immunomodulatory effects of its co-chaperonin, hsp10, in experimental arthritis. Our results show that an aqueous solution of a mycobacterial hsp10 delayed the onset and severity of adjuvant-induced arthritis in rodents when administered after disease induction but before joint involvement occurred. This biological activity was specific for the hsp10 of Mtb, since neither GroES nor the rat homologue was effective. Using synthetic hsp10 fragments, the activity was localized to the N-terminal region of the molecule. Assessment of circulating antibody levels to mycobacterial hsp10 and hsp65 indicated that all arthritic rats had increased litres to both hsp10 and hsp65: hsp10-treated rats showed further elevation of this humoral response not only to hsp10 but also to hsp65 when compared with the untreated arthritic control. This is the first report of the immunomodulatory activity of mycobacterial hsp10 in experimental arthritis, and exhibits a potential role for this co-chaperonin in pathophysiological situations.     

Egr-1基因剔除减少炎性相关因子在实验性胰腺炎中的表达

目的：探讨Egr-1基因剔除对实验性胰腺炎小鼠胰腺组织中炎性相关因子表达的影响。 〖HTH〗方法：利用Egr-1基因剔除小鼠，采用大剂量雨蛙素诱导的实验性胰腺炎模型，观察Egr-1基因剔除后，胰腺组织水肿、MPO水平、血清淀粉酶水平、肺组织MPO水平的改变。并利用定量PCR的方法，检测胰腺组织中炎症相关因子组织因子（TF）、纤维蛋白溶酶原激活因子抑制因子（PAI-1）、单核细胞趋化吸引蛋白1（MCP-1）、Gro-1、IL-6和细胞间黏附分子-1（ICAM-1） mRNA的表达。 〖HTH〗结果：Egr-1基因剔除小鼠胰腺组织水肿明显轻于野生型，但组织MPO水平与血清淀粉酶与野生型组间相比无明显差异；肺组织MPO水平明显低于野生型。定量PCR检测结果表明， Egr-1基因剔除组，胰腺组织中TF、PAI-1，以及MCP-1、ICAM-1和IL-6　mRNA的表达，明显少于野生型组。 〖HTH〗结论：Egr-1基因剔除可明显减轻急性胰腺炎的严重程度，其作用可能通过减少胰腺组织中TF、PAI-1，以及MCP-1、ICAM-1和IL-6的表达而实现。     

中药“神经再生素”促神经生长过程中基因差异性表达的初步研究

为了探索中药“神经再生素”促神经生长过程中基因水平的变化。本实验采用 dd-PCR方法 ,从体外培养背根神经节细胞加药组和不加药组中获得两者的差异表达片段 ,并经反杂交筛选、克隆测序、DNA序列检索分析、Northern验证。结果表明 ,差异显示共获得 8个差异条带 ,一个为下调基因 ,其余为上调基因 ,其中 2个 c DNA序列与 RRAJ5 16 1基因 (增殖相关基因 )、AF196 3 15基因 (锌指样蛋白 DDP2 ) 10 0 %同源 ,2个 c DNA序列与 AK0 0 175 7基因、STA5 SRR基因 (t RNA合成酶 )部分同源。结论是中药“神经再生素”在促神经生长过程中 ,对神经元基因的选择性表达起着重要的调控作用。     

Roles of tumour necrosis factor-alpha (TNF-alpha), transforming growth factor-beta (TGF-beta), and IL-10 in the modulation of intercellular adhesion molecule-1 (ICAM-1) expression by macrophages during mycobacterial infection

Profiles of ICAM-1 expression on cultured murine peritoneal macrophages infected with Mycobacterium avium complex (MAC) were examined, with special reference to modulating roles of TNF-alpha, TGF-beta, and IL-10. When macrophages were infected with MAC, ICAM-1 expression, measured by microscopic counting of ICAM-1+ macrophages stained with anti-ICAM-1 antibody, ELISA, and flow cytometric analysis, was rapidly increased, peaking at day 3 (early-phase up-regulation) due to endogenous TNF-alpha, and thereafter gradually declined to the normal level within 1 week or more (late-phase down-regulation). The late-phase ICAM-1 down-regulation was also seen in macrophages phagocytosing heat-killed MAC and those stimulated with lipopolysaccharide but not in macrophages phagocytosing latex beads. ICAM-1 mRNA expression was augmented markedly at day 1 after MAC infection and thereafter decreased. While TNF-alpha and IL-10 production by MAC-infected macrophages was observed during the first 3 days, TGF-beta production was initiated from day 3 and continued until day 14. Exogenously added TGF-beta strongly inhibited the early-phase increase in ICAM-1 expression by infected macrophages, and the blockade of endogenous TGF-beta with anti-TGF-beta antibody markedly inhibited late-phase ICAM-1 down-regulation. Moderate blocking effect was also observed for anti-IL-10 antibody. On the other hand, late-phase ICAM-1 down-regulation was not prevented by the addition of exogenous TNF-alpha. Therefore, TGF-beta and IL-10, especially the former, appear to play active roles in the late-phase down-regulation of ICAM-1 in MAC-infected macrophages during long-term cultivation.     

Production of IL-6 by T cells from the femoral head of patients with rapidly destructive coxopathy (RDC)

RDC is a syndrome with unknown etiology that causes rapid destruction of a hip joint. We have investigated the production of osteoclast-activating cytokines (IL-6, IL-1α and tumour necrosis factor-alpha (TNF-α)), interferon-gamma (IFN-γ) and IL-8 by T cells in the affected joint. The level of IL-6 produced by the T cell lines (TCL) established from the femoral head was significantly higher than that from patients' or healthy donors' peripheral blood mononuclear cells (PBMC). IL-6 production by the TCL from synovial membrane or from patients' PBMC was also significantly higher than that from healthy donors' PBMC. IL-1α production by the TCL from the femoral head was significantly higher than any of the other groups when all the TCL were used for the analysis. TNF-α production was highest in the TCL from patients' PBMC. The levels of IFN-γ or IL-8 were not significantly different among these four groups. The plasma levels of all these cytokines except for IFN-γ, that was rather lower, in RDC patients were not significantly different from those in osteoarthrosis or trauma patients, or healthy donors. These results suggest that T cells at the affected femoral head, and also synovial membrane to some extent, are involved in bone resorption through the production of IL-6 and probably IL-1α in patients with RDC.     

Immunopathological features of palatine tonsil characteristic of IgA nephropathy: IgA1 localization in follicular dendritic cells

IgA nephropathy (IgAN) is generally thought to be mediated by the glomerular deposition of circulating immune complexes containing IgA as the major antibody component. Upper respiratory infections and tonsillitis often precede IgAN. and in some cases tonsillectomy is affective for the (treatment of IgAN. Thus, the tonsil seems to be a unique organ causing initial and/or progressive events to generate nephritogenic immune complexes in IgAN. in this study we focused on the analysis of immunopathological features of the palatine tonsil characteristic of IgAN patients by using an immunohistochemical technique. The IgAl subclass was demonstrated in follicular dendritic cells (FDC) of the tonsil of IgAN patients, but not in FDC of non-IgAN controls. On the other hand, IgA2, IgG, IgM and C3 did not show any differences in distribution between the two groups. Moreover, the expression of decay-accelerating factor (DAF), an inhibitor of homologous complement activation, and transforming growth factor-beta I (TGF-/β1). an inducer of antibody-producing ceils to IgA class switching, in FDC and interdigitating dendritic cells of the tonsil, respectively, which was also clarified in this study for the first time, was found to be identically distributed in the two groups. These findings may support the idea that IgA1. possibly in an immune complex form, is trapped by FDC and plays an important role in the persistent activation of particular B cell repertoires responsible for ihe onset and/or progression of IgAN.     

Cloned cellular reagents to define antigens encoded between HLA-DR and glyoxylase

Primed lymphocyte typing reagents have been used to define antigens encoded by genes of a locus (loci) mapping between HLA-DR and glyoxalase I. This locus, which we shall refer to as the third locus of the HLA-D region, has been variously referred to as D beta, PL beta, PL3, and SB. Generating discriminatory primed lymphocyte typing reagents which can be used to define these antigens, however, has been extremely difficult. Donors of responding and stimulating cells for the priming combinations have usually been matched not only for the DR, D, and MB/MT antigens but also for the HLA-A, -B, and -C antigens. Even under these very restricted conditions, not all bulk primed lymphocyte typing reagents that are generated are discriminatory enough to be useful for antigen definition. We have derived "clones" from bulk priming combinations in which stimulator and responder differed for known antigens of this third locus. Even though the bulk reagents that were prepared did not provide discriminatory results, approximately 7-12% of the clones derived from the bulk priming combination proved to be highly discriminatory. We have been able to obtain these results with regard to all three antigens of the third locus so far evaluated. The very ease of screening clones and deriving discriminatory reagents, as compared with screening responder-stimulator combinations, allows the ready derivation of cellular reagents that define the antigens of this third locus.