纯系LEW大鼠及裸小鼠膀胱灌注给药方式的探讨

目的探讨纯系LEW大鼠及裸小鼠膀胱灌注给药方法.方法显微镜下解剖雌性LEW大鼠及裸小鼠尿道并根据解剖特征改造套管针.通过插入尿道的套管针膀胱灌注墨水,同时观察插管及膀胱充盈和染色情况.持续观察小鼠和大鼠生存状况及尿道损伤情况.结果显微镜下可见雌性LEW大鼠及裸小鼠尿道与人存在明显差异.经尿道插管成功率高(100%),动物生活状况没有明显受到插管灌注影响.结论裸鼠膀胱灌注给药治疗是一种成功率高而可行的给药手段,动物膀胱灌注化疗模型可能为研究膀胱肿瘤治疗提供更为有效的工具.     

经尿道灌注自杀基因治疗大鼠膀胱癌的实验研究

目的:探讨经尿道膀胱灌注脂质体和逆转录病毒载体介导的自杀基因(HSVtk),观察在羟基无环鸟苷(ganciclovir,GCV)作用下大鼠膀胱肿瘤的生长情况.方法:用N-甲基-亚硝基脲(MNU)膀胱灌注建立大鼠原位膀胱癌模型.经尿道膀胱灌注脂质体、逆转录病毒载体HSVtk基因,随后腹腔内注射GCV,连续6天.结果:经尿道膀胱灌注HSVtk基因后48h,RT-PCR检测出膀胱肿瘤中HSVtk基因表达.逆转录病毒组和脂质体组膀胱总重量与裸质粒组和对照组相比有显著差异,表明膀胱肿瘤的生长被抑制.TUNEL法原位检测肿瘤组织中凋亡细胞不仅出现在表层细胞,而且深层细胞大量凋亡,推测旁观者效应可能发挥了一定作用.结论:经尿道膀胱灌注脂质体DNA或重组逆转录病毒载体可以向肿瘤转导HSVtk基因.HSVtk/GCV系统能抑制大鼠原位膀胱肿瘤的生长,显示出抗肿瘤作用.     

浅表性膀胱癌模型的建立及其活体荧光成像

目的:建立浅表性膀胱癌模型并以活体荧光成像系统观察肿瘤生长情况.方法:利用脂质体将增强型绿色荧光蛋白(EGFP)表达质粒转染人膀胱癌细胞株KU-7,G418筛选稳定表达EGFP的克隆.采用经尿道膀胱灌注法使肿瘤细胞种植于膀胱黏膜.活体荧光荧光成像系统直接观察肿瘤细胞生长及形成瘤体的过程.结果:建立了转染率接近100%的人膀胱癌KU-7/EGFP细胞系,在体外及裸小鼠体内均能够长期稳定表达绿色荧光蛋白.活体荧光成像观察发现,1～4周随着肿瘤体积逐渐增大,肿瘤的发光随着观察时间的延长而增加.结论:利用绿色荧光蛋白标记膀胱肿瘤细胞,建立一种浅表性膀胱癌模型,为连续动态实时观察和准确评价自然状态下肿瘤细胞生长过程提供了新的手段.     

经尿道电气化术加灌注化疗治疗多发膀胱肿瘤42例报告

目的探讨经尿道膀胱肿瘤电气化术结合灌注化疗治疗多发性膀胱肿瘤的方法及疗效。方法应用经尿道电气化术切除多发性膀胱肿瘤42例。术后第1周开始用丝裂霉素C（MCC）20mg灌注膀胱,每周1次,共6次,然后每4周1次,持续1年以上。结果手术时间平均45min,未发生大出血及膀胱穿孔。平均随访16个月,术后复发9例,复发率21．4％。结论经尿道膀胱肿瘤电气化术结合灌注化疗治疗多发性膀胱肿瘤是一种有效的方法。     

术中膀胱黏膜下注射盐酸氮芥、术后即刻丝裂霉素灌注化疗预防膀胱癌复发

目的探讨经尿道膀胱肿瘤电切(TUR-Bt)术中膀胱黏膜下注射盐酸氮芥、术后即刻丝裂霉素(MMC)膀胱灌注化疗预防膀胱癌复发的疗效。方法 112例行TUR-Bt术的膀胱癌患者随机分为2组,对照组56例:行TUR-Bt术后1周给予MMC常规膀胱灌注化疗;治疗组56例:TUR-Bt术中经尿道行膀胱黏膜下注射盐酸氮芥,术后再即刻行MMC膀胱灌注化疗,术后1周再给予MMC常规膀胱灌注化疗。比较观察2组患者术后3、6、12、18、24个月的复发情况。结果治疗组术后3、6、12、18、24个月的复发率低于对照组,比较差异有统计学意义(P<0.05)。结论 TUR-Bt术中膀胱黏膜下注射盐酸氮芥、术后即刻行MMC膀胱灌注化疗能显著提高膀胱癌治疗效果,减少术后复发,且无明显毒副反应。     

序贯治疗告知单联合电话随访对膀胱肿瘤患者术后治疗依从性的影响

目的探讨应用序贯治疗告知单联合电话随访对膀胱肿瘤患者术后膀胱灌注化疗依从性的影响。方法将78例膀胱肿瘤患者按查随机数字表法分为观察组和对照组，两组各39例。观察组给予发放序贯治疗告知单及电话随访进行干预，对照组给予口头健康教育。观察两组患者按时返院行膀胱灌注化疗和遵嘱饮水的依从性，以及膀胱刺激征的发生情况等。结果观察组第1阶段遵嘱饮水及第2阶段、第3阶段按时返院行膀胱灌注化疗的依从性均高于对照组（P〈0．05），观察组第1阶段膀胱刺激征的发生率明显低于对照组（P〈0．05）。结论序贯治疗告知单联合电话随访可提高膀胱肿瘤患者术后膀胱灌注化疗及遵嘱饮水的依从性，降低膀胱刺激征的发生率。     

小剂量丝裂霉素C膀胱内灌注预防浅表性膀胱癌术后复发的临床观察

目的 评价小剂量丝裂霉素膀胱内序列灌注预防浅表性膀胱癌术后复发的疗效及安全性。方法 对1998年至2004年42例浅表性膀胱癌患者术后进行膀胱内灌注化疗，其中32例行经尿道膀胱肿瘤(TURBt)电切术，10例行膀胱部分切除术，所有入选病例均经病理证实为浅表性肿瘤，术后1周开始应用丝裂霉C20mg加蒸馏水40mL，1次／周，6次后改为每月1次，维持1年。结果 所有入选病例均获随访12～24个月，12个月内肿瘤复发3例，复发后进展2例，38例随访24个月无复发。42例中均未见明显药物副作用，仅有4例出现轻微尿道刺激症状。结论 丝裂霉素C对浅表性膀胱癌有较明显的治疗和预防复发的作用，灌注后副作用小，患者耐受性好，是较好的预防和治疗浅表膀胱癌的药物。     

膀胱癌电气化术后膀胱内灌注吡柔比星预防复发的疗效(附37例报告)

目的　评价吡柔比星 (THP)膀胱内灌注预防膀胱癌电气化术 (TUV Bt)后复发的疗效及安全性。方法　对 37例浅表性膀胱癌患者行经尿道膀胱肿瘤电气化术 ,术后定期应用THP(30mg/ 30ml)膀胱内灌注 (保留 30分钟 ) ,观察其疗效及安全性。结果　 37例全部随访 2～ 18个月 ,无 1例复发。除 2例创面不愈合并发溃疡外 ,无明显不良反应。结论　THP膀胱内灌注 ,初步疗效表明其预防膀胱肿瘤术后复发效果显著 ,长期疗效待进一步观察。     

MNU诱导的近交系大鼠膀胱肿瘤模型

目的通过膀胱灌注N-甲基亚硝基脲(MNU)诱导近交系大鼠膀胱肿瘤。方法选取F344大鼠20只，膀胱灌注MNU，每2周1次，共5次；于第14周处死动物，取膀胱肿瘤组织测MHC—Ⅰ及病理观察。结果MNU组肿瘤发生率为100％；膀胱肿瘤的MHC—Ⅰ表达较高(58．60％～93．80％)。结论MNU可成功诱导F344大鼠膀胱肿瘤的形成，是肿瘤免疫治疗的理想模型。     

全反式维甲酸联合干扰素治疗膀胱癌

 目的　观察联合应用全反式维甲酸（ATRA）和干扰素α-2a（IFN-α-2a）治疗大鼠膀胱肿瘤的疗效并探讨其机制。方法　建立大鼠膀胱肿瘤模型50只，44只存活的大鼠随机分为A（11只）、B（11只）、C（11只）、D（11只）4组，膀胱内分别灌注DMSO、IFN-α-2a、ATRA和ATRA+IFN-α-2a治疗。显微镜观察病理分期及分级，流式细胞术（FCM）检测肿瘤细胞的凋亡和细胞周期。结果　D组大鼠体重（277.4±18.5）g高于其他3组，膀胱重量（0.19±0.05）g低于其余3组，并且D组病理分期和分级均有显著降低。D组肿瘤细胞凋亡率（13.43±3.06）％较其他三组明显增加，而增生指数（19.95±4.17）％则明显降低。结论　ATRA和IFN-α-2a联合应用能显著促进肿瘤细胞凋亡，对膀胱肿瘤有更好的疗效。     

联合灌注预防浅表性膀胱肿瘤术后复发的临床观察

 目的 了解联合膀胱腔内灌注化疗方案对预防浅表性膀胱癌复发的疗效，探讨一种理想的膀胱腔内灌注方法。方法 将45例(男性34例，女性11例)年龄在31～80岁(平均56．1岁)浅表性膀胱癌患者随机分为A、B和C3组。A组接受单次表阿霉素灌注作为对照，B、C组接受不同的羟基喜树碱和丝裂霉素C序贯膀胱腔内灌注联合用药方案。结果 A、B、C组平均随访时间分别为26．6月、36．73月、24．31月，复发率分别为54．5％(6／11)、27．7％(5／18)、6．25％(1／16)。结论 羟基喜树碱和丝裂霉素C合理地联合应用能取得较好疗效。     

Growth of murine sarcoma virus-transformed rat kidney cells in nude mice: absence of induction of host endogenous viruses.

Clonal isolates of the normal rat kidney cell line (NRK) transformed by a defective murine sarcoma virus (Kirsten strain) were injected into nude mice of BALB/c background to determine whether the growth of these cells as tumors was accompanied by the induction of host endogenous type C viruses. All the virus-transformed clones produced rapidly growing tumors in nude mice, but neither the induction of mouse endogenous viruses nor the rescue and spread of the transforming sarcoma virus were observed during the growth of tumors. The degree of expression of the tumor virus structural proteins in the transformed cells did not determine the cellular phenotype with regard to tumorigenicity in nude mice, nor did it modify the cellular growth properties in vitro. Consistent with earlier observations with simian virus 40-transformed mouse and rat cells, the ability of sarcoma virus-transformed NRK cells to initiate tumor growth in nude mice appeared to be correlated with anchorage-independent growth in vitro.     

GFP image analysis in the mouse orthotopic bladder cancer model

Precise and objective measurements of tumor response have yet to be standardized in the mouse orthotopic bladder cancer model. In this study, we used image analysis and green fluorescent protein (GFP) to objectively measure tumor size in response to chemotherapy. KU-7 human bladder cancer cells transfected with GFP were intravesically inoculated into 8-week-old female nude mice. Fourteen days after tumor cell inoculation, the mice were assigned into a control (PBS) group or a doxorubicin (conc. 1.0 mg/ml) treatment group and received a single instillation of treatment. Fourteen days after treatment, the bladders were surgically exposed and fluorescent images were captured and later analyzed using image analysis. Bladders were processed for histological examination. Tumor incidence determined by GFP expression and histology was 100 and 80%, respectively, in the doxorubicin treatment group. A 9-fold (histology) vs. 12-fold (GFP expression) difference in tumor regression measured by tumor area (P<0.05) and a 5-fold (histology) vs. 9-fold (GFP expression) difference in tumor regression measured by the percent of tumor area in the bladder (P<0.001) were observed in the doxorubicin treatment group. Our findings suggest that using image analysis provides a precise, sensitive and objective means to measure tumor growth and treatment response in the mouse orthotopic bladder cancer model in lieu of histological methods. Consequently, the number of mice required in an experiment can be reduced since tissue samples are not needed for histology, thus making tissue samples readily available for additional assays in both a labor-effective and cost-effective manner.     

C-Type virus particles in a cell line from a lymphosarcoma of a nude mouse.

C-type virus particles were observed by electron microscopy in cultures of a cell line (NML-1) derived from a lymphosarcoma that arose spontaneously in a nude NIH outbred mouse. The presence of such particles indicated that human tumors heterotransplanted in nude mice might become contaminated with murine oncornaviruses.     

Acinar cell carcinoma of rat pancreas: regulation of cholesterol esterification

The regulation of cholesterol esterification during cell proliferation was studied. The serum free cholesterol, cholesterol esters and lecithin: cholesterol acyltransferase (LCAT) activity of nude mice with and without pancreatic acinar cell tumours and rats with proliferating tissues were determined. In addition, the apparent activity of acyl-CoA: cholesterol acyltransferase (ACAT) in homogenates of nude mouse tumours and proliferating rat tissues were determined and compared with those of normal nude mouse and rat tissues. Serum cholesterol ester levels were significantly lower in host nude mice with tumours and in rats with regenerating liver, and increased significantly in pregnant rats when compared with respective controls. Circulating LCAT activity levels decreased in host nude mice, in pregnant rats, and in rats with regenerating pancreas and regenerating liver. Apparent ACAT activity levels increased significantly in nude mouse tumours and in foetal and postnatal rat pancreata and also in postnatal liver. At the same time, apparent ACAT activity levels decreased in foetal and regenerating rat livers when compared with respective control tissues. These results suggest that serum cholesterol esters, circulating LCAT and cellular ACAT levels are modulated during cell proliferation.     

白血病细胞来源的树突状细胞在治疗白血病中的应用

目的:探讨免疫毒素BDI-1-PEA特异杀伤膀胱癌细胞的体外及体内研究.方法:应用MTT法检测不同浓度的BDI-1-PEA,BDI-1和PEA的混合物(BDI-1 PEA),PEA对体外癌细胞的杀伤能力.接种于裸鼠背部皮下的癌细胞长至0.3cm大小实体瘤时,于尾静脉隔日分别注入BDI-1-PEA、BDI-I、正常小鼠IgG共6次,观察不同的药物对癌的抑制作用.结果:免疫毒素BDI-1-PEA在体外抗膀胱癌细胞系E-J的作用明显优于PEA及BDI-1与PEA的混合物,在荷瘤裸鼠体内明显优于BDI-1及IgG,与对非靶细胞的细胞毒性作用相比较差异非常显著.结论:免疫毒素BDI-1-PEA是一种很有潜力的抗癌药物,为进一步临床研究奠定基础.     

Growth of human digestive-tumour xenografts in athymic nude rats.

The athymic nude rat rnu/rnu has been established as an in vivo model for the acceptance of human digestive-tumour xenografts. We report the successful xenografting of 7/12 (58%) primary explants from patients with digestive cancer. Successful xenografting also occurred in 21/25 (84%) pancreatic tumours derived from a pancreatic exocrine adenocarcinoma (GER) maintained in cell culture; 2 of those have been successfully passaged in nude rats. The simultaneous implantation of these tumours into nude mice led to an almost identical take rate. Passage of one colonic and one pancreatic xenograft from nude rats into nude mice, and transplantation back into nude rats, increased the take rates. The critical period for the establishment of primary tumour growth was usually 28-42 days. The xenografts maintained histological and cytological characteristics of the primary explants or of the original tumour from which the cell line derived. The karyotype of the cell line was also maintained in the solid tumour. Three murine tumours were successfully grown as xenografts. Despite their immunoincompetence, the rats in this study showed no increased morbidity or mortality when kept in conventional conditions, compared with animals housed in isolators. The athymic nude rat will become a valuable complementary tool to the nude mouse for the establishment and maintenance of human digestive tumours and for surgical and serial serological studies.