To establish an economical and reproducible method for the high-quality RNA extraction from pancreas, we isolated total RNA from rat pancreas with TRIzol

The black seeds, from the Ranunculaceae family, have been traditionally used by various cultures as a natural remedy for several ailments. In this study, we examined the effect of black seed oil as an immunomodulator in a rat model of allergic airway inflammation. Rats sensitized to ovalbumin and challenged intranasally with ovalbumin to induce an allergic inflammatory response were compared to ovalbumin-sensitized, intranasally ovalbumin-exposed rats pretreated with intraperitoneally administered black seed oil and to control rats. The levels of IgE, IgG1 and ova-specific T-cell proliferation in spleen were measured by ELISA. The pro-inflammatory cytokine IL-4, IL-5, IL-6 and TGF-b1 mRNA expression levels were measured by reverse transcription polymerase chain reaction. The intraperitoneal administration of black seed oil inhibited the Th2 type immune response in rats by preventing inflammatory cell infiltration and pathological lesions in the lungs. It significantly decreased the nitric oxide production in BALF, total serum IgE, IgG1 and OVA-specific IgG1 along with IL-4, IL-5, IL-6 and TGF-b1 mRNA expression. Black seed oil treatment resulted in decreased T-cell response evident by lesser delayed type hypersensitivityand lower T-cell proliferation in spleen. In conclusion, black seed oil exhibited a significant reduction in all the markers of allergic inflammation mainly by inhibiting the delayed type hypersensitivity and T-cell proliferation. The data suggests that inhibition of T-cell response may be responsible for immunomodulatory effect of black seed oil in the ratmodel of allergic airway inflammation.

基于复杂性疾病的临床异质性，或称表型异质性;遗传异质性，或称位点异质性;多基因的微效作用;异位显性（epistasis）;拟表型(phenocopy);环境因素的作用等特点，鉴定复杂性疾病的易感基因是一种艰巨有时难以完成的任务。近交系动物遗传背景清晰，动物实验可控制环境因素的干扰，以及转基因动物在研究基因功能中独特的作用，应用动物模型为筛选鉴定人类复杂性疾病的易感基因提供了不可替代的工具。以大鼠类风湿关节炎模型为例，定位克隆易感基因步骤包括: (1)选择亲代近交系, 建立关节炎模型; (2)分离育种，连锁分析、确定数量性状位点(QTLs);(3)建立Congenic系、窄化QTL区域;(4)位置克隆基因;(5)易感基因的功能验证。我们应用这一策略从朴日斯烷(Pristane)诱导的关节炎大鼠模型中已筛选出十余个控制关节炎发生发展的非MHC基因位点，在此基础上克隆的Pia4基因被鉴定为是中性粒细胞胞浆因子1（Ncf1），说明此一研究策略是完全可行的，并且可能发现新的功能基因或已知基因的新功能。

Type II collagen (CII) is a target for autoreactive Tcells in both rheumatoid arthritis and the murine model collagen-induced arthritis. The determinant core of CII has been identified as CII260-270, and the alteration of this T cell epitope by posttranslational modifications is known to be critical for development of arthritis in mice. Using CIIspecific Tcell hybridomas we have now shown that the immunodominant Tcell epitope in the normal (healthy) human and rat joint cartilage is O-glycosylated at the critical T cell receptor recognition position 264 with a mono- or di-saccharide attached to a hydroxylysine. In contrast, in the arthritic human and rat joint cartilage there are both glycosylated and non-glycosylated CII forms. Glycosylated CII from normal cartilage could not be recognized by T cells reactive to peptides having only lysine or hydroxylysine at position 264, showing that antigen-presenting cells could not degrade the O-linked carbohydrate. Thus, the variable forms of the glycosylated epitope are determined by the structures present in cartilage, and these vary during the disease course. We conclude that the chondrocyte determines the structures presented to the immune system and that these structures are different in normal versus arthritic states.