Application of Serum-Free Culture Medium for Preparation of A-NK Cells

To compare the differences between proliferation and cytotoxicity of adherent natural killer （A-NK） cells cultured with serum-free medium AIMV and standard serum-containing medium in vitro, and also observe the assisting effect of IL-12 on the activation and the morphology character of IL-2-treated A-NK cells, cellular proliferation was evaluated by MTT method in vitro. The morphology of the target cells killed by A-NK cells was observed through electroscope. All of the A-NK cells cultured in serum-free medium AIMV could rapidly proliferate and keep high cytotoxicity compared with that in standard serum-containing medium. A-NK cells activated by both moderate-dose IL-2 and IL-12 were superior to the high-dose IL-2-treated A-NK cells. These data indicated that serum-free medium AIMV could replace standard serum-containing medium for culturing A-NK cells, and moderate-dose IL-2 and IL-12 could reduce side effects caused by high-dose IL-2. The study provided a new experimental basis for experimental and clinical preparation of A-NK cells. Cellular ＆ Molecular Immunology.     

Characterization of mouse allergens

The major allergens present in mouse skin, serum, and urine have been identified. Skin extracts, serum, and urine were chromatographed, and the activities of the fractions were monitored by histamine release from the leukocytes of individuals sensitive to mice. Fractionation of skin extracts revealed two major allergens. The large allergen has a molecular weight of approximately 67,000 daltons and by biochemical and immunochemical criteria appears to be identical to mouse albumin. The smaller molecular weight allergen is approximately 17,000 daltons. The same two allergens are also found in mouse serum and mouse urine. Histamine release by leukocytes of individuals allergic to mice demonstrated that some individuals react predominantly to the large allergen, some to the small allergen, and one group of patients reacts to both allergens.     

Circulating levels of placental protein 12 and chorionic gonadotrophin following RU 38486 and Gemeprost for termination of first trimester pregnancy

First trimester termination of pregnancy was successfully inducedin ten patients with RU 38486 followed 2 days later by a prostaglandin(Gemeprost) pessary. Human chorionic gonadotrophin (HCG) valuesremained unaltered until after the abortion. The levels of placentalprotein 12 (PP12) showed an immediate and significant fall followingRU 38486, then rose to values substantially higher than thoseat the initial visit after 2 days. These findings show thatRU 38486 has a direct inhibitory effect on tissues producingPP12 and confirm the progesterone dependency of this protein     

gp130胞外区近膜端缺失的突变体介导IL-6信号的研究

目的研究ｇｐ１３０胞外区远膜端和近膜端在ＩＬ－６信号转导中的作用。方法用分子克隆手段构建全长的膜结合型ｇｐ１３０分子和胞外区第３０１～５８２位氨基酸残基缺失的突变体分子，并通过阻滞电泳方法比较突变体与野生型分子在传导信号方面的差异。结果通过酶切鉴定、序列分析和表达检测，证明野生型和突变体ｃＤＮＡ表达载体构建正确，并有效表达；阻滞电泳结果表明，在ＳＫＯ－００７细胞和Ｍｏｌｔ－４细胞中，突变体分子与野生型ｇｐ１３０都能介导ＩＬ－６信号。结论ｇｐ１３０分子胞外区远膜端在ＩＬ－６信号转导中发挥重要作用；而近膜端并非ＩＬ－６信号转导所必需。     

IL-12 inhibits in vitro immunoglobulin production by human lupus peripheral blood mononuclear cells (PBMC)

Systemic lupus erythematosus (SLE) is a prototypical autoimmune disease characterized by polyclonal B cell activation and by the production of anti-double-stranded (ds) DNA antibodies. Given the inhibitory effects of IL-12 on humoral immune responses, we investigated whether IL-12 displayed such an activity on in vitro immunoglobulin production by SLE PBMC. Spontaneous IgG, IgG1, IgG2, IgG3 and IgM antibody production was dramatically reduced by addition of IL-12. These results were confirmed by Elispot assays detecting IgG- and anti-dsDNA-secreting cells. While IL-6 and TNF titres measured in PBMC supernatants were not modified by addition of IL-12, interferon-gamma (IFN-γ) titres were up-regulated and IL-10 production down-regulated. Since addition of IFN-γ did not down-regulate immunoglobulin production and since the inhibitory activity of IL-12 on immunoglobulin synthesis was not suppressed by anti-IFN-γ antibody, we concluded that the effect of IL-12 on immunoglobulin production was not mediated through IFN-γ. Our data also argue against the possibility that down-regulation of endogenous IL-10 production was responsible for the effect of IL-12. Thus, inhibition of IL-10 production by IFN-γ was not accompanied by inhibition of immunoglobulin production, and conversely, restoration of IL-10 production by anti-IFN-γ antibody did not suppress the inhibitory activity exerted by IL-12 on immunoglobulin production. Taken together, our data indicate that reduction of excessive immunoglobulin and anti-dsDNA antibody production by lupus PBMC can be achieved in vitro by IL-12, independently of IFN-γ and IL-10 modulation.     

The COX-2 inhibitor, rofecoxib, ameliorates dextran sulphate sodium induced colitis in mice

Objective: We have evaluated the efficacy of the selective cyclo-oxygenase (COX)-2 inhibitor, rofecoxib, for the prevention of experimental colitis.Material and methods: To induce colitis BALB/c mice received 5% dextran sulphate sodium (DSS) in their drinking water continuously for 7 days. Rofecoxib (2.5–10 mg/kg body weight, p.o.) was administered throughout the treatment period with DSS. Colitis was quantified by a clinical damage score, colon length, weight loss, stool consistency and rectal bleeding. Inflammatory response was assessed by neutrophil infiltration, determined by histology and myeloperoxidase (MPO) activity. Interleukin (IL)-1, prostaglandin (PG)E₂ and PGD₂ levels in colon mucosa and the immunohistochemical expression of COX-1 and –2 were also studied.Results: The COX-2 inhibitor ameliorated severe colitis, reduced the degree of inflammation through reduction of neutrophil infiltration and IL-1 levels. PGE₂, and PGD₂ synthesis were significantly reduced in DSS-treated groups. Indeed, treatment with rofecoxib diminished the lost of COX-1 caused by DSS in the crypt epithelium whereas expression of COX-2 remained unaffected.Conclusions: Rofecoxib is protective in acute DSS – induced colitis, probably by reducing neutrophil infiltration, inhibiting up-regulation of IL-1 and returning to normal COX-1 expression in the inflamed colonic mucosa.Received 19 April 2004; returned for revision 17 June 2004; accepted by I. Ahnfelt-Rønne 23 November 2004     

模拟腰椎前路、后路间盘摘除术T12-L5腰段脊柱应力松弛蠕变实验研究

研究了18例新鲜尸体T12－L5腰段脊柱应力松弛，蠕变特性。测定了完整脊柱（正常组）及模拟前路（对照1组），后路手术（对照2组）腰段脊柱的应力松弛和蠕变效应，得出了在恒应变，应力条件下应力－时间曲线及数据，用回归分析的方法处理实验数据，得出了归一化应务松弛，蠕变函数及曲线，对前路间盘摘除术与后路间盘摘除术对脊柱粘弹性的影响进行分析讨论。