异种肝移植免疫排斥反应中脾脏的作用

目的 探讨脾脏在仓鼠到大鼠异种肝移植中的体液免疫,细胞免疫作用。方法 三袖套法建立仓鼠到大鼠原位肝移植模型,观察移植术后肝脏和脾脏的病理变化;采用补体介导的细胞毒检测异种肝移植术后血浆中抗仓鼠抗体滴度的变化;运用免疫组织化学的方法检测脾脏中大鼠抗体的产生,脾脏增殖细胞核抗原的表达;并观察环孢素Ａ,环磷酰胺,切脾对移植术后生存时间及免疫排斥反应的影响。结果 异种肝移植术后免疫排斥反应发生时,脾脏急剧     

Successful Xenograft of Endoscopic Ultrasound-Guided Fine-Needle Aspiration Specimen from Human Extrahepatic Cholangiocarcinoma into an Immunodeficient Mouse

Patient-derived tumor xenograft is the transfer of primary human tumors directly into an immunodeficient mouse. Patient-derived tumor xenograft plays an important role in the development and evaluation of new chemotherapeutic agents. We succeeded in generating a patient-derived tumor xenograft of a biliary tumor obtained by endoscopic ultrasound-guided fine-needle aspiration from a patient who had an inoperable extrahepatic cholangiocarcinoma. This patient-derived tumor xenograft will be a promising tool for individualized cancer therapy and can be used in developing new chemotherapeutic agents for the treatment of biliary cancer in the future.     

人结肠癌裸鼠移植瘤细胞动力学与雌孕激素受体研究

将人结肠癌异种移植于ＢＡＬＢ／ｃｎｕ／ｎｕ系无胸腺裸鼠并传５代。对原结肠癌及各代移植瘤进行病理学、组化、细胞动力学与电镜检查。应用流式细胞仪（ＦＣＭ）与生化葡聚糖包裹活性碳吸附法（ＤＣＣ）对各代移植瘤进行胞核ＤＮＡ倍体与雌孕激素受体（ＥＲ·ＰＲ）检测，并与原人结肠癌比较，结果显示：各代移植瘤均保持原人结肠癌相同的病理学特征、阳性雌孕激素受体表达及稳定的胞核ＤＮＡ倍体。就受体作用失常参与大肠癌发病机制进行讨论，为动态观察性激素受体变化规律及从实验药理学探讨内分泌治疗的真正价值提供依据     

B‐Cell Depletion Extends the Survival of GTKO.hCD46Tg Pig Heart Xenografts in Baboons for up to 8 Months

Xenotransplantation of genetically modified pig organs offers great potential to address the shortage of human organs for allotransplantation. Rejection in Gal knockout (GTKO) pigs due to elicited non‐Gal antibody response required further genetic modifications of donor pigs and better control of the B‐cell response to xenoantigens. We report significant prolongation of heterotopic alpha Galactosyl transferase “knock‐out” and human CD46 transgenic (GTKO.hCD46Tg) pig cardiac xenografts survival in specific pathogen free baboons. Peritransplant B‐cell depletion using 4 weekly doses of anti‐CD20 antibody in the context of an established ATG, anti‐CD154 and MMF‐based immunosuppressive regimen prolonged GTKO.hCD46Tg graft survival for up to 236 days (n = 9, median survival 71 days and mean survival 94 days). B‐cell depletion persisted for over 2 months, and elicited anti‐non‐Gal antibody production remained suppressed for the duration of graft follow‐up. This result identifies a critical role for B cells in the mechanisms of elicited anti‐non‐Gal antibody and delayed xenograft rejection. Model‐related morbidity due to variety of causes was seen in these experiments, suggesting that further therapeutic interventions, including candidate genetic modifications of donor pigs, may be necessary to reduce late morbidity in this model to a clinically manageable level.     

caveolin-1抑制乳腺癌细胞体内外增殖及其机制探讨

目的研究caveolin-1对乳腺癌细胞MCF7在体内、体外生长和增殖的影响并初步探讨其机制。方法选用人类乳腺癌细胞MCF7,通过基因转染技术培育过表达caveolin-1的细胞株MCF7/cav-1作为实验组,空载体细胞株MCF7/vec作为对照组。Western blot方法检测转染细胞内caveolin-1和血管内皮生长因子（VEGF）的表达。软琼脂集落形成实验检测细胞增殖克隆的能力。建立裸鼠皮下种植瘤模型并检测肿瘤组织中Ki-67与VEGF的表达情况。结果 caveolin-1在MCF7/cav-1细胞中表达稳定,其表达量明显高于对照组细胞MCF7/vec（P〈0.01）和亲本细胞MCF7（P〈0.01）,而对照组细胞MCF7/vec和亲本细胞MCF7之间caveolin-1表达量差异无统计学意义（P〉0.05）。与MCF7/vec细胞比较,MCF7/cav-1细胞在软琼脂中形成的集落数目明显减少（P〈0.01）,体积也较小。MCF7/cav-1细胞中VEGF表达量明显低于MCF7/vec细胞（P〈0.01）。在裸鼠的体内实验中,与MCF7/vec细胞比较,MCF7/cav-1细胞在裸鼠体内生长缓慢（P〈0.01）,Ki-67染色明显减弱,VEGF染色阴性,提示caveolin-1可以抑制肿瘤体内增殖。结论 caveolin-1可能通过抑制VEGF表达抑制MCF7细胞生长和增殖。     

Advanced glycation end products-mediated hypertrophy is negatively regulated by tetrahydrobiopterin in renal tubular cells

Adrenocortical carcinoma (ACC) is a very rare but aggressive tumor, whose biological and cellular features and processes underlying the development, progression and metastatic evolution are still obscure. Despite many attempts to use general cytostatic and cytotoxic drugs, the only available drug therapy for advanced ACC is still represented by mitotane (MTT). However, the mechanism of action of this adrenolytic derivative of the pesticide DDT has still been poorly characterized. In this context, the development of more specific drugs for ACC treatment is based on the knowledge of the molecular pathways involved in the tumor growth. Xenograft models for the screening of such drugs at preclinical levels is mandatory. In the first part of this review, we will summarize the "pro" and "con" of the different xenograft models available for anticancer drug testing in different types of tumors in general and in the last part, we will focus on the preclinical evidence obtained so far with the use of such models applied to drug screening for anticancer effects in ACC.     

鱼油组分共轭亚油酸抑制肺腺癌干细胞成瘤的体内研究

目的:建立肺腺癌干细胞荷瘤动物模型,体内验证鱼油组分共轭亚油酸(CLA)抑制肺腺癌增殖及转移的作用和分子机制. 方法:采用新霉素和克隆挑选技术分别建立稳定表达绿色荧光蛋白(GFP)的A549和SPC-A-1细胞株,体外培养富集干细胞微球,建立肺癌干细胞原位荷瘤裸鼠模型,腹腔注射给予不同组分c9t11-CLA、tl0c12-CLA和混合组,定期检测裸鼠体重和肿瘤质量,研究终点采用小动物活体荧光成像技术分别观察肿瘤的转移情况.采用Western blotting检测肿瘤组织COX-2、CXCR4的蛋白表达水平. 结果:实验终点各组原发肿瘤重量,A549混合治疗组比各单药治疗组肿瘤质量显著降低(P＜0.001),而各CLA单药治疗组比,A549 control组肿瘤质量显著降低(P＜0.01),与SPC-A-1组结果相似.各组肿瘤转移情况:在动物活体荧光成像系统下,A549CSC和SPC-A-1 CSC空白对照组出现了全身多处转移;经c9,t1 1-CLA、t10,c12-CLA治疗显著减少转移位置和比率;而混合治疗组减少尤为明显.Western blotting检测OCT4、COX-2、CXCR4蛋白表达水平.在A549肺腺癌干细胞组,c9t11-CLA、t1oc12-CLA均不同程度地抑制OCT4、COX-2、CXCR4蛋白,混合治疗组抑制作用最强.在SPC-A-1肺腺癌干细胞组结果类似. 结论:c9,t1 1-CLA、t10,c12-CLA及其混合物能明显抑制肺腺癌干细胞荷瘤裸鼠原发肿瘤和转移灶的生长;CLA组分能明显抑制肿瘤组织OCT4、COX-2和CXCR4蛋白的表达.     

1H-MRS活体检测rAd/p53治疗人类结肠癌荷瘤裸鼠肿瘤代谢物变化

目的运用磁共振氢质子频谱(Proton MRS,1H-MRS)分析研究rAd/p53治疗人类结肠癌荷瘤裸鼠的早期改变,探索1H-MRS活体无创评价rAd/p53早期抗癌疗效的标记物。方法 50只人类结肠癌SW480荷瘤BALB/c裸小鼠随机分成两组:对照组,rAd/P53治疗组。于假治疗或治疗后24h、48h、72h、120h、168h各组分别随机取5只荷瘤鼠行1H-MRS。随后处死荷瘤鼠,即切下肿瘤,检测肿瘤的p53蛋白表达(Western blot)、凋亡(TUNNEL)。结果 24h:CHO/CR面积比及GLX β～γ/CR面积比较对照组减少(P<0.05);48h:Lip峰高及面积较对照组增加(P<0.05);72h以上:GLX β～γ/CR面积比较对照组减少(P<0.05);120h以上:GLXα/CR面积比较对照组减少(P<0.05)。三个治疗组在观察时间点肿瘤坏死、凋亡范围较对照组增加,p53蛋白表达均较对照组增高。CR面积、Lip峰高和面积与肿瘤细胞凋亡呈正相关;GLX β～γ/CR的面积比、GLX α/CR的面积比与肿瘤细胞凋亡呈负相关。结论 rAd/p53治疗肿瘤早期CHO/CR、GLXα/CR与GLX β～γ/CR面积比出现降低,Lip出现增加,与肿瘤细胞的凋亡增加相关,可作为肿瘤治疗早期疗效评价的生物标志物。