微囊化大鼠胰岛异种移植治疗小鼠实验性糖尿病的研究

目的 研究海藻酸钠－聚赖氨酸－海藻酸钠包裹胰岛进行移植的效果。方法 将Ｗｉｓｔａｒ大鼠的胰腺先行胶原酶胰管内注射消化,然后分离,纯化,所得胰岛经培养后制成微囊包膜的胰岛,微囊直径为０．４￣０．５ｍｍ,每个微囊内包１个胰岛。     

人CD59/MCP双基因对人血灌注转基因小鼠离体心脏功能的保护作用

目的研究转人补体调节蛋白hCD59/hMCP双基因抑制人补体激活从而抑制补体介导的超急性反应、延长体外灌注小鼠心脏功能的作用。方法利用显微注射法建立转hCD59/hMCP双基因小鼠的模型,其子代中hCD59单基因阳性小鼠8只、hMCP单基因阳性小鼠11只和hCD59/hMCP双基因阳性小鼠8只分别为3个实验组,同窝非转基因小鼠10只为对照组。用新鲜人血浆体外灌注小鼠心脏,研究小鼠表达抑制人补体激活的人补体调节蛋白后,对补体介导的异种移植超急性排斥反应的抑制作用。结果转双基因组心脏搏动时间(138min±25min)明显长于单基因组(hCD59组78min±27min,hMCP组43min±21min)及非转基因组(20min±12min)(Dunnett′s T检验,均P<0.01);免疫荧光染色及免疫酶组织化学染色显示转双基因组心脏内补体C3c及C9沉积也明显减少。结论转人补体调节蛋白hCD59/hMCP双基因能够有效地抑制补体介导的异种移植超急性排斥反应。     

异种肝细胞移植排斥机理的探讨

目的：探讨异种肝细胞排斥反应的机理,为治疗异种肝细胞移植排斥反应提供理论依据。方法,用D－氨基半乳糖腹腔内注射制成肝功能衰竭大鼠模型,并在其脾内植入豚鼠肝细胞,通过免疫组化方法,用抗大鼠IgM抗体和抗大鼠IgG抗体,抗大鼠CD4和抗大鼠CD8抗体与因排斥反应在大鼠脾 内可能产生的IgM,IgG抗体和CD4,CD8淋巴细胞结合,在移植后不同时间取受鼠脾脏标本,检测其内是否有IgM,IgG抗体和CD4,CD8淋巴细胞存在。结果：移植后12hIgM抗体存在,移植后24h大鼠脾内出现IgG抗体,同时有CD4^=和CD8^＋淋巴细胞,且在移植后1周大鼠脾内均可见上述4种物质。结论：体液和细胞免疫均参与异种肝细胞移植的排斥反应。     

腹腔与静脉应用托泊替坎治疗人卵巢癌细胞株SKOV3裸小鼠网膜移植瘤的疗效比较

Objective:To compare the therapeutic and toxic profile of topotecan given intraperitoneally with intravenously in human ovarian cancer xenografted into athymic nude mice.Methods: Eighty female Balb-c/nu-nu mice were randomized assigned into eight groups (n=10). Xeneografts resulted from intramesentery injection of cultured human ovarian cancer cells SKOV3 in athymic mice. Onset of intraperitoneal treatment with either topotecan or cisplatin (7.5 mg/kg) was on day 7. Animals scheduled for topotecan i.p. received intraperitoneal application of topotecan (1.5 mg/kg×2, 3.0 mg/kg×2, 6.0 mg/kg×2 or 10.0 mg/kg×1). Animals scheduled for topotecan i.v. received intravenous administration of topotecan (6.0 mg/kg×2 or 10.0 mg/kg×1). Two weeks after drug application animals were killed. Tumor growth inhibition were assessed and compared with untreated mice and cisplatin intraperitoneally administered mice. Acute toxicity was determined by loss of body weight. Cell cycle division and apoptosis after drug administration was determined by flow cytometric analysis.Results: In a panel of ten tumour xenografts, intraperitoneal topotecan was significantly more effective than intravenous administration. The toxicity profile suggested a better tolerability in terms of weight loss after intraperitoneal administration than cisplatin control. Topotecan 10.0 mg/kg i.p. per day (1 day) schedule was an optimal treatment for ovarian cancer and well tolerated by mice with no signs of acute toxicity. Topotecan and cisplatin induce cells G0-G1 arrest and apparent apoptosis. No significant difference among mice treated with topotecan intraperitoneally or intravenously or cisplatin was observed in term of apoptosis and cell cycle perturbation.Conclusion:The results may have implications for the future design of clinical studies on intraperitoneal application of topotecan. It suggests that apoptosis and cell cycle perturbation play an limited role in the mechanism of topotecan administration.     

胎儿静脉动脉化肾上腺异体移植的应用解剖

对40具7个月以上新鲜胎儿的左肾上腺静脉和左肾静脉的起始、走行、外径、长度及汇入部位进行了观测,以提供胎儿静脉动脉化肾上腺异体移植的有关形态学资料。基于大网膜呼吸能力较强和有适合吻合的动脉,认为其是理想的移植部位。     

Recombinant Bordetella pertussis pertactin (P69) from the yeast Pichia pastoris: high-level production and immunological properties

Acellular whooping cough vaccines are based on pertussis toxoid but their effectiveness may be increased by the addition of other Bordetella pertussis antigens. We expressed the immunogenic outer membrane protein pertactin (P69) from B. pertussis to high levels in multi-copy transformants of the industrial yeast Pichia pastoris. In high-density fermentations, engineered P. pastoris yielded > 3 g of the protein per litre of culture. Purified recombinant pertactin was able to stimulate the incomplete protection afforded by toxoid to the level of the whole-cell vaccine, as shown by the Kendrick test, supporting its inclusion in future acellular vaccines.     

异种脱钙骨基质,异种骨基质胶加释放系统移植的组织学观察

Ｗｉｓｔａｒ大鼠２４只分４组，分别将４种复合异种骨植入大鼠股四头肌内，定期取材制片，光镜及ＴＥＭ－１０ＳＸ透射电镜观察。结果：２ｄ巨噬细胞等密集成团，细胞器发达；４ｄ间充质细胞伪足伸入异种脱钙骨基质（Ｄｅｍｉｎｅｒａｌｉｚｅｄ ｘｅｎｏｇｅｎｅｉｃ ｂｏｎｅ ｍａｔｒｉｘ，ＸＤＢＭ），异种骨基质胶（Ｘｅｎｏｇｅｎｅｉｃ ｂｏｎｅ ｍａｔｒｉｘ ｇｅｌａｔｉｎ，ＸＢ－ＭＧ）隙内；８ｄ间充质细胞增多，     

Cyclic AMP-mediated regulation of vascular smooth muscle cell cyclic AMP phosphodiesterase activity

1. Rat cultured aortic vascular smooth muscle cells (VSMC) express both cyclic GMP-inhibited cyclic AMP phosphodiesterase (PDE3) and Ro 20-1724-inhibited cyclic AMP phosphodiesterase (PDE4) activities. By utilizing either cilostamide, a PDE3-selective inhibitor, or Ro 20-1724, a PDE4-selective inhibitor, PDE3 and PDE4 activities were shown to account for 15% and 55% of total VSMC cyclic AMP phosphodiesterase (PDE) activity.
2. Treatment of VSMC with either forskolin or 8-bromo-cyclic AMP caused significant concentration- and time-dependent increases in total cellular cyclic AMP PDE activity. Using cilostamide or Ro 20-1724, we demonstrated that both PDE3 and PDE4 activities were increased following forskolin or 8-bromo-cyclic AMP treatment, with a relatively larger effect observed on PDE3 activity. The increase in cyclic AMP PDE activity induced by forskolin or 8-bromo-cyclic AMP was inhibited by actinomycin D or cycloheximide, demonstrating that new mRNA synthesis and protein synthesis were required. An analogue of forskolin which does not activate adenylyl cyclase (1,9-dideoxyforskolin) or an analogue of cyclic GMP (8-bromo-cyclic GMP) did not affect total cyclic AMP PDE activity.
3. Incubation of VSMC with 8-bromo-cyclic AMP for 16 h caused a marked rightward shift in the concentration-response curves for both isoprenaline- and forskolin-mediated activation of adenylyl cyclase. A role for up-regulated cyclic AMP PDE activity in this reduced potency is supported by our observation that cyclic AMP PDE inhibitors (IBMX, cilostamide or Ro 20-1724) partially normalized the effects of isoprenaline or forskolin in treated cells to those in untreated cells.
4. We conclude that VSMC cyclic AMP PDE activity is increased following long-term elevation of cyclic AMP and that increases in PDE3 and PDE4 activities account for more than 70% of this effect. Furthermore, we conclude that increases in cyclic AMP PDE activity contribute to the reduced potency of isoprenaline or forskolin in treated VSMC. These results have implications for long-term use of cyclic AMP PDE inhibitors as therapeutic agents.