供者特异性抗原诱导异种免疫耐受的初步研究

目的：为探讨异种免疫耐受的可能性。方法：以豚鼠－大鼠心脏移植建立超急排斥反应模型,在眼镜蛇毒因子（ＣＶＦ,３０Ｕ／ｋｇ）及环磷酰胺（ＣｙＰ）的覆盖下,将供者特异性或非特异性血管内皮细胞以４．０×１０６个／只,注入受者的门静脉,２周后行颈部心脏移植。结果：移植供心存活时间获得延长至（４６．１±５）ｍｉｎ,加用ＣｙＰ后延长尤为明显（５４．５±９．７ｍｉｎ）,同时血管内皮细胞溶解率下降,ＣｙＰ对移植物存活也有一定的延长作用（３４．６±８．７ｍｉｎ）,而非特异性血管内皮细胞则无此作用。结论：在豚鼠－大鼠异种移植模型中,供者特异性血管内皮细胞经门静脉径路注射可诱导受者产生一定程度的免疫耐受。这种耐受的产生机制尚需进一步研究。     

Photodynamic therapy using methylene blue in lung adenocarcinoma xenograft and hamster cheek pouch induced squamous cell carcinoma

BackgroundPhotodynamic therapy (PDT) is used to treat early proximal bronchial cancer during a flexible bronchoscopy. The technique relies on the excitation of a photosensitizer by an appropriate wavelength, which is delivered into the bronchus in close contact with the tumor.ObjectiveTo assess methylene blue (MB) as a PDT agent for the treatment of respiratory tract cancer in animal models.MethodsMB-induced PDT was performed on 7 subcutaneous NCI-H460 lung adenocarcinoma xenografts in nude mice and 9 induced squamous cell cancer in the hamster cheek pouch model. In mice, PDT was carried out on right-sided tumors after intratumoral injection of methylene blue 1% (w/v) and illumination at 630 nm at 200 J/cm (Diomed PDT 630), with the left tumor used as control (illumination alone or MB alone). The tumoral volume was assessed before and 15 days after PDT.ResultsFourteen xenografts were treated in mice, including seven treated with MB-PDT, producing a 52% mean tumor volume regression (1568 mm³ vs. 544 mm³) compared to seven control cases in which tumor volume increased (p = 0.007; Mann-Whitney test). Nine cheek pouch induced carcinomas were treated in the hamster group, with a mean volume decrease of 85.8% (from 44.8% to 100%) (initial mean volume = 210 mm³ vs. post PDT mean volume = 97 mm³). Histology analysis showed 4/9 complete responses.ConclusionIntratumoral MB appears efficient as PDT agent for cancer treatment in animal models. Further studies are needed to assess the safety and efficacy of MB-associated PDT for the treatment of lung cancer in humans.     

Transplantable zebrafish models of neuroendocrine tumors

Neuroendocrine tumors (NETs) are rare neoplasms whose incidence is increasing. NETs constitute a heterogeneous group of tumors. Their clinical features, functional properties, and clinical course are different on the basis of their site of origin. Due to the heterogeneity of these tumors, a coordinated multidisciplinary approach is required in these patients. However, medical doctor encounters many difficulties when providing care for patients with NETs. This review provides an overview of the state of the art of zebrafish model in the cancer research with a main focus on NETs.     

受体外周血IgG、MΦ、NK、CD4、CD8及MLR的变化与异种移植物存活的关系

目的探讨受体外周血IgG、MΦ、NK、CD4、CD8及MLR的变化与异种移植物存活的关系。方法选用小鼠移植于大鼠耳后心肌组织模型，按实验所设Ⅰ、Ⅱ、Ⅲ、Ⅳ组在术前12、8、4、0d及10、6、2、0d分别将小鼠脾细胞1×10     

Diclofenac potentiates the antitumor effect of cisplatin in a xenograft mouse model transplanted with cisplatin-resistant cells without enhancing cisplatin-induced nephrotoxicity

Cisplatin (CDDP) is a well-known anticancer agent, and CDDP–induced nephrotoxicity (CIN) is one of the most serious adverse effects. Previously, we revealed that while celecoxib reduces CIN, diclofenac does not appear to enhance it. Furthermore, we reported that diclofenac additively enhances the cytotoxic effect of CDDP on CDDP-resistant A549 cells (A549/DDP cells) and their spheroids. In addition, celecoxib reduces the cytotoxic effect of CDDP on A549/DDP cells while demonstrating an anticancer effect; however, it enhanced the effect of CDDP cytotoxicity on spheroids. Therefore, we evaluated the effects of diclofenac or celecoxib on CIN and the antitumor effect of CDDP in a xenograft mouse model transplanted with A549/DDP cells. Although CDDP did not decrease tumor size and tumor weight, these parameters were significantly reduced following co-administration with diclofenac when compared with the control group. Conversely, celecoxib marginally suppressed the antitumor effect of CDDP. Moreover, CDDP increased the mRNA levels of kidney injury molecule 1 (Kim-1), a renal disorder marker, in the kidneys of xenograft mice; treatment with celecoxib and diclofenac did not impact Kim-1 mRNA levels increased by CDDP. In conclusion, diclofenac potentiated the antitumor effect of CDDP without enhancing CIN.     

CD19基因过表达K562细胞株及NOD-SCID小鼠皮下移植瘤模型的建立

目的 建立人CD19基因过表达的K562细胞株(CD19-K562)NOD-SCID小鼠皮下移植瘤模型.方法 克隆人CD19基因构建MigR1-CD19反转录病毒载体,转入Plat-A包装细胞,病毒上清转染K562细胞,反转录聚合酶链反应(PCR)及流式细胞术检测转染细胞CD19基因和蛋白表达,体外反复传代获得CD19-K562稳定转染细胞株,锥虫蓝染色,细胞计数检测细胞增殖,Annexin V/碘化丙啶(PI)双染流式细胞术检测细胞凋亡.CD19-K562细胞株皮下接种NOD-SCID小鼠并经体内外传代后建立移植瘤亚系CD19-K562-a,建立NOD-SCID小鼠皮下移植瘤模型.应用反转录PCR、瑞特染色、免疫组织化学等方法验证CD19-K562-a细胞性质.结果 成功建立了人CD19-K562细胞稳定株,CD19阳性率(99.80±0.17)％.CD19-K562细胞增殖、凋亡未受转染及传代影响.CD19-K562细胞稳定株皮下接种NOD-SCID小鼠后经体外结合体内传代建立移植瘤亚系CD19-K562-a,其CD19阳性率为(99.78±0.04)％,CD19-K562-a和CD19-K562细胞均呈未分化形态.CD19-K562-a皮下接种NOD-SCID小鼠成功建立移植瘤模型,CD19-K562-a瘤体CD19基因表达强阳性.结论 通过构建MigR1-CD19重组载体获得CD19基因过表达的K562稳定转染细胞株(CD19-K562),同时采用体外结合体内传代建立稳定成瘤的NOD-SCID小鼠移植瘤亚系CD 19-K562-a并成功建立NOD-SCID小鼠皮下移植瘤模型.     

人源肿瘤异种移植模型的研究进展

肿瘤细胞系培养技术的出现有力地推动了肿瘤生物学研究地发展,但在肿瘤新疗法地临床转化研究中,细胞系肿瘤模型地预测能力有限,且不稳定.人源肿瘤异种移植模型能高度保留原肿瘤内基质异质性、组织学特性和分子多样性,引起研究者关注并被广泛应用于肿瘤新疗法研究,有望为肿瘤患者个体化治疗带来新突破.     

以海藻酸盐及异种骨复合技术制备组织工程化骨及体内成骨

背景：海藻酸有相对温和的凝胶条件与良好的生物相容性,已广泛应用于生物组织工程。 目的：采用海藻酸钠凝胶复合异种骨的方法,构建骨组织工程载体,观察载体中细胞的生物性能及体内成骨能力。 方法：取2 只2 周龄新西兰兔的骨髓,以1×10-8 mol/L重组人骨形态发生蛋白2诱导骨髓间充质干细胞。取诱导后第2 代骨髓间充质干细胞接种于1%海藻酸钠凝胶中,培养4 d 苏木精-伊红染色观察凝胶中细胞形态。将第2 代骨髓间充质干细胞分为单纯DMEM凝胶组和含1%海藻酸钠的DMEM凝胶组,分别培养7 d 后行骨形态发生蛋白2免疫组织化学染色观察。取24 只裸鼠,随机分为2 组,于两侧股部肌袋中分别植入骨髓间充质干细胞/海藻酸钠凝胶/牛松质骨复合体作为实验组,骨髓间充质干细胞/牛松质骨复合体作为对照组。术后2,4 周后组织学观察复合体成骨情况,图像分析系统分析各组成骨或软骨的面积百分比。 结果与结论：海藻酸钠凝胶中骨髓间充质干细胞形态饱满,细胞悬浮于凝胶中,可见细胞分裂和核分裂相。单纯DMEM凝胶组和含1% 海藻酸钠的DMEM凝胶组免疫组织化学观察,细胞分裂增殖正常,伸出多种形态的突起,胞核大,核仁清晰。单纯DMEM凝胶组和含1%海藻酸钠的DMEM凝胶组的骨形态发生蛋白2 表达阳性率差异无显著性意义(P > 0.05)。扫描电镜观察海藻酸钠凝胶均匀地复合于牛松质骨微孔中,不同平面均有细胞生长。动物实验显示术后2,4 周实验组和对照组的成骨或软骨的面积百分比差异有显著性意义（P < 0.05）。提示以海藻酸钠凝胶/牛松质骨构建骨组织工程载体,合乎组织工程载体的超结构原理,能最大限度地承载细胞,生物性能好,对骨髓间充质干细胞增殖和成骨表型及相关的生物性能无不良影响,在体内成骨效率较高。 关键词：组织工程骨;海藻酸钠凝胶;异种骨;载体;支架材料 doi:10.3969/j.issn.1673-8225.2010.08.043