目的 观察肝大部切除大鼠再生血清和肝细胞生长因子（HGF）诱导骨髓干细胞向肝实质细胞分化的作用；探讨骨髓干细胞向肝细胞分化和增殖的体外培养条件和方法，为患者应用自身骨髓细胞修复损伤肝脏提供实验依据。方法 制作肝大部切除大鼠肝再生动物模型，收集术后24h血清；分别应用鼠肝再生血清和HGF对大鼠骨髓细胞进行定向诱导分化培养；以免疫组化、RT-PCR和westernblot等方法对分化不同阶段细胞进行检测。将分化细胞经尾静脉输入同系大鼠，观察输注细胞在肝、脾内的生长情况。结果 活体移植分化细胞能定向迁移至肝和脾；体外及体内分化细胞在mRNA水平和蛋白贡水平均表达白蛋白，并在一定时期内表达甲胎蛋白。结论 大鼠骨髓干细胞在肝大部切除再生血清或HGF的诱导下能横向分化为肝实质样细胞。

Prostate-specific membrane antigen (PSMA) is a cell surface glycoprotein expressed predominantly in prostate secretory acinar epithelium and prostate cancer cells as well as in several extraprostatic tissues. Mouse monoclonal antibody 4G5 specific to the extracellular domain of PSMA was used to screen two phage displayed peptide libraries (9aa linear and 9aa cys library). Three 4G5-reactive phagotopes were identified. Sequence analysis of isolated clones demonstrated that the interaction motif VDPA/SK has high homology to 719-725aa on PSMA. Immunohistochemical staining of the prostate cancer sample with the PSMA-mimic phagotope (mimotope) immunized serum antibodies demonstrate that the mimotope isolated from the phage displayed peptide libraries can induce PSMA specific immune response in vivo.

Pertussis toxin (PTX) has been widely used as an adjuvant to induce Th1-mediated organ-specific autoimmune diseases in animal models. However, the cellular and molecular mechanisms remain to be defined. In this study, we showed that dendritic cells (DC) stimulated with PTX (PTX-DC) were able to substitute for PTX to promote experimental autoimmune uveitis (EAU). EAU induced by PTX-DC revealed a typical Th1 response, characterized by high uveitogenic retinal Ag interphotoreceptor retinoidbinding protein (IRBP)-specific IFN-and IL-12 production in the draining lymph nodes, as well as increased levels of anti-IRBP IgG2a and decreased levels of anti-IRBP IgG1 in the serum of IRBP-immunized mice. Furthermore, PTX-DC preferentially induced T cells to produce the Th1 cytokine, IFN-. After being stimulated with PTX, DC exhibited up-regulation of MHC class II, CD80, CD86, CD40, and DEC205. PTX-DC had also increased allostimulatory capacity and IL-12 and TNF-production. Serum IL-12 was increased in naive mice that received PTX-DC i.p. In addition, PTX activated extracellular signal-regulated kinase in DC. Following the inhibition of extracellular signal-regulated kinase signaling, the maturation of PTX-DC was reduced. Subsequently, the ability of PTX-DC to promote IFN-production by T cells in vitro and to induce EAU in vivo was blocked. The results suggest that PTX might exert an adjuvant effect on DC to promote their maturation and the production of proinflammatory cytokines, thereby eliciting a Th1 response.

We investigated whether recipient dendritic cells (DCs), pretreated with nuclear factor-kB oligodeoxyribonucleotide decoy (NF-kB ODN decoy) and loaded with ultraviolet B-irradiated donor apoptotic splenocytes (Apo-SCs), were able to induce murine cardiac allograft tolerance. Heterotopic vascularized heart transplantation was performed from BALB/c to C57BL/6 mice, and recipients (C57BL/6) were given one injection of recipient DCs pretreated with NF-kB ODN decoy and loaded with donor (BALB/c) apoptotic SCs (decoy Apo-SCs DCs) through the portal vein at 7 days, before heart transplantation in the absence of immunosuppression. The cardiac allograft survival time and the expressive levels of intragraft cytokine genes [interleukin (IL)-2, IL-10, and interferon-γ] were evaluated. Our results indicated that injection of decoy Apo-SCs DCs significantly prolonged vascularized heart allograft survival and led to skewing of intragraft cytokine expression towards T helper 2 (IL-10). The mechanisms can be useful for therapy of allograft rejection with minimization of systemic immunosuppression.

目的 构建含小鼠次级淋巴组织趋化因子（SLC）cDNA的重组腺相关病毒载体。方法 以C57BL/6J小鼠的淋巴组织为材料抽提总RNA，用RT-PCR方法克隆小鼠SLC的成熟肽基因。PCR产物回收后经EcoRI和Xho双酶切，插人pAAv-IRES-hrTGFP质粒，用磷酸钙法，与pAAV-Rc和pHetpeR三者共同转染HEK293细胞。包装成重组的病毒颗粒，并通过绿色荧光蛋白（CFP）报告基因荧光检测及抽提病毒DNA，进行PCR扩增等进一步证实重组病毒的形成。结果 约500bp的小鼠SLC成熟肽基因被成功克隆，序列分析表明，所克隆的SLC基因与基因库注册的相同，组载体的酶切及测序鉴定表明SLC基因被定向插入。重组载体转染HEK293细胞，经荧光显微镜和病毒DNA的PCR检测，证实已完成对重组病毒的包装。并表达GEP。结论 成功构建了SLC成熟肽基因重组腺相关病毒载体。