Frequency of T-Cell Progenitors in Nude Mice

The hypothesis that prothymocytes are distinct from and regulated independently of multilineage hemopoietic progenitors was tested by enumeration of these two cell populations in normal versus congenitally athymic (nude) mice. The absence of a thymus and of peripheral T cells in nude mice had no effect on the frequency of either multilineage progenitors (day 12 CFU-S) or prothymocytes (CFU-T), suggesting that there is no feedback regulation of CFU-T frequency. Thymus seeding from the bone marrow is therefore likely to be regulated by the availability of niches for prothymocyte maturation, rather than by feedback control of prothymocyte production.     

Enzyme-linked immunosorbent assay for Giardia-specific IgA in mouse intestinal secretions

We describe an ELISA for trophozoite-specific IgA in the intestinal secretions of mice infected with Giardia muris. Using this method, trophozoite-specific IgA was demonstrated in intestinal secretions of Giardia-infected immunocompetent BALB/c mice. Such antibody was undetectable in the intestinal secretions of Giardia-infected athymic (nude) mice. Immunocompetent BALB/c mice are able to clear G. muris infection whereas nude mice are not. The study provides evidence that the chronicity of G. muris infection in nude mice results from lack of intestinal trophozoite-specific IgA in these animals. By means of the ELISA, trophozoite-specific IgA was demonstrated in intestinal secretions from immunocompetent mice in the absence of protease inhibitors.     

抗端粒酶核酶抑制肝癌细胞裸鼠移植瘤生长的研究

设计针对端粒酶RNA组分的特异性核酶，研究该核酶对肝癌细胞裸鼠移植瘤生长的影响。构建含针对端粒酶RNA组分锤头状核酶基因的重组真核表达质粒pBBS212-RZ，以及建立人肝癌细胞株HepG2裸鼠移植瘤模型。将不同剂量的pBBS212-RZ用脂质体包裹后进行瘤体内多点注射，对照组注射生理盐水或空质粒载体pBBS212。连续注射2l天后，测量移植瘤体积和重量，检测瘤组织端粒酶活性。抗端粒酶特异性核酶使瘤组织端粒酶活性下降(抑制率为72％)，明显地抑制了肝癌细胞裸鼠移植瘤生长(抑制率为68％)，且存在剂量依赖性。抗端粒酶特异性核酶作为一种有效的端粒酶抑制剂，可抑制肝癌细胞的生长，有望在肝癌基因治疗中发挥作用。     

Experimental chronic dermatophytosis in human skin grafted to nude mouse: inoculation of Trichophytons and histopathological evaluation

Two strains of T. rubrum and one strain of T. mentagrophytes were inoculated into human skin grafted onto BALB/c nude mice by the needle puncture method. Infection was established in 1 of the 10 animals inoculated with fluffy colony type T. rubrum, 2 of the 10 animals inoculated with powdery colony type T. rubrum, and 7 of the 10 animals inoculated with granular colony type T. mentagrophytes, suggesting that the skin grafts are infectible by anthropophilic and zoophilic strains of dermatophytes. T. rubrum infection continued for a maximum of 9 weeks and T. mentagrophytes infection for more than 11 weeks. In the animals inoculated with T. mentagrophytes, fungal elements were localized in the stratum corneum of the human skin grafts. In the acute stage, microabscesses consisting of neutrophils were observed under the stratum corneum in contact with fungal elements; in the chronic stage, epidermal thickening and infiltration, mainly consisting of histiocytes and a smaller number of lymphocytes, was noted in the upper and middle dermis. Ultrastructural findings from the parasites were similar to those of dermatophytosis in man. This experimental system should be useful as a model of chronic dermatophyte infection in the human skin.     

Enhanced in vivo cytotoxicity of recombinant human tumor necrosis factor with etoposide in human renal cell carcinoma

Summary The combination of tumor necrosis factor (TNF) and etoposide (ETP) was evaluated for potential cytotoxic efficacy against a human renal cell carcinoma xenograft using an in vivo assay employing an athymic mouse host with tumor implanted a the subrenal capsule site. Both antitumor efficacy (relative survival or RTS) and toxicity (weight loss) of TNF and ETP alone and in combination were evaluated. While TNF and ETP alone were mildly inhibitory (RTS 90% and 71%, respectively), the combination caused marked tumor inhibition (45% of controls). Host toxicity encountered with the combination did not exceed the toxicity associated with ETP alone, suggesting that the therapeutic index may have been augmented. It is concluded that enhanced antitumor activity without substantial augmentation of toxicity is observed with this combination, providing a rationale for further evaluation of tumor necrosis factor-based regimens for the treatment of advanced renal carcinoma.Supported by a Merit Review grant, VA Medical Research Service, Durham, NC 27710, USA     

Metastatic Behavior and Tumorigenicity of a Human Melanoma Cell Line (MM-RU) after Injection into Nude Mice

The ability of the human amelanotic melanoma cell line MM-RU to produce experimental metastases and to grow tumors at subcutaneous inoculation sites in 4-week-old nude mice was examined. After i.v. inoculation of 10⁶ cells, all injected mice (n=21) developed consistent numbers of metastatic pulmonary colonies within 32 days. The coefficients of variation for the number of colonies were between 17%–23% in three independent experiments. Survival time after i.v. inoculation was 63 ± 7 days (mean ± SD) (n=20). Within 20 days, subcutaneous inoculation of 5 × 10⁶ cells resulted in tumor growths of 13 ± 3 mm (mean ± SD) at the inoculation sites in all nude mice (n=12). The MM-RU cell line seems to be a simple, fast vehicle for testing the effect of melanoma growth modulators on experimental pulmonary metastases as well as on subcutaneously growing melanoma.     

荷人鼻咽癌裸鼠血浆MDA,cAMP／cGMP水平及PSP的影响

本实验利用ＢＡＬＢ／Ｃ裸小鼠，复制荷人鼻咽癌裸鼠模型，检测荷瘤鼠血浆丙二醛，ｃＡＭＰ，ｃＧＭＰ和ｃＡＭＰ／ｃＧＭＰ比值，观察云芝糖肽对荷人ＮＰＣ裸鼠的抑瘤作用及对荷瘤鼠血浆ＭＤＡ和ｃＡＭＰ／ｃＧＭＰ等的影响。结果发现荷瘤鼠血浆ＭＤＡ升高，ｃＡＭＰ，ｃＧＭＰ降低，ｃＡＭＰ／ｃＧＭＰ比值升高，高你低浓度ＰＳＰ对荷人ＮＰＣ裸鼠均有明显抗癌作用（Ｐ＜０．０１），抑瘤率５９．５８－９５．５３％；并可降低荷瘤     

Pituitary Folliculo-Stellate-Like Cells Stimulate Somatotroic Pituitary Tumor Growth in Nude Mice

A new cell line (TtT/GF) established from a murine pituitary thyrotropic tumor having characteristics similar to those of pituitary folliculo-stellate cell (FS cell) was implanted into nude mice together with cells from a rat pituitary somatotrophic tumor cell line (MtT/S) to determine whether the former enhances pituitary tumor growth. For as long as 2-3 mo after implantation, MtT/S cells implanted either alone or together with fibroblasts formed either no tumors or only very small tumors in the nude mice. In contrast all of the nude mice that had received MtT/S cells implanted together with TtT/GF cells developed large tumors. Furthermore, the mice bearing the MtT/S and TtT/GF implants showed a significantly higher body weight and serum growth hormone level than those bearing only MtT/S cells or a combination of MtT/S cells and fibroblasts. The TtT/GF cell line itself had no tumorigenicity during the experimental period. Therefore, the TtT/GF cell line as a model of FS cells enhanced pituitary endocrine cell tumor formation. Additionally, immunocytochemistry showed that TtT/GF cells positive for glial fibrillary acidic protein (GFAP) or S-100 protein were present in the parenchymatous tissue elements or connective tissue surrounding the tumor nests. In the parenchymatous tissue, the TtT/GF cells exhibited a stellate appearance and surrounded neighboring tumor cells with their long cell processes. These results suggest that TtT/GF cells can serve as a model for pituitary FS cells, and are capable of stimulating pituitary tumor growth either by modifying the microenvironment or producing growth factors.     

肺癌靶向多肽荧光分子探针的研制及其应用

目的:选择能与整合素α3受体结合的小分子多肽cNGQGEQc-L作为靶向载体，将羧基荧光素（FAM）与cNGQGEQc-L连接构建荧光分子探针，通过荧光成像探讨荧光多肽分子探针用于肺腺癌显像的可行性。方法:利用倒置荧光显微镜观察FAM-cNGQGEQc-L与肺腺癌A549细胞结合部位，流式细胞仪检测荧光多肽与A549细胞的竞争抑制实验，观察FAM-cNGQGEQc-L随浓度的增加与肺腺癌A549细胞结合能力的变化情况。通过小动物活体成像仪，观察荧光多肽在荷瘤裸鼠体内的生物分布特点。结果:倒置荧光显微镜显示荧光多肽cNGQGEQc-L能与A549细胞结合，结合部位在细胞膜和细胞质中。流式细胞仪测试结果证明荧光多肽与A549细胞的结合具有特异性和饱和性，当FAM-cNGQGEQc-L浓度为0.125 mmol/L时，荧光多肽与A549细胞的结合趋近饱和。荷瘤裸鼠活体成像显示移植瘤能够摄取荧光多肽，且荧光多肽通过泌尿系统和胆道系统排泄。结论:体外、体内荧光实验结果显示，荧光多肽分子探针FAM-cNGQGEQc-L可与肺腺癌A549细胞、肺癌移植瘤结合，能够特异性靶向肺腺癌。     

皮肤涂抹雌三醇对裸鼠甲状腺功能和肿瘤影响的效果观察

目的研究经皮涂抹外源性雌三醇(E₃)对裸鼠甲状腺功能及甲状腺移植瘤的影响。方法使用123只4周龄裸鼠,按体质量采用随机数字表法分为9组:①对照组、②低E₃组、③高E₃组、④乳头状甲状腺癌组(PTC组)、⑤乳头状甲状腺癌+高E₃组(PTC+高E₃组)、⑥滤泡状甲状腺癌组(FTC组)、⑦滤泡状甲状腺癌+高E₃组(FTC+高E₃组)、⑧未分化型甲状腺癌组(ATC组)和⑨未分化型甲状腺癌+高E₃组(ATC+高E₃组),均正常饮食饮水。除对照组、PTC组、FTC组和ATC组外,其余5组每日给予皮肤涂抹雌三醇乳膏,低E₃浓度为市售化妆品人均每日使用量中雌三醇含量的25倍、高剂量为100倍。40天后,收集各组裸鼠的24h尿样;尿碘采用砷铈催化分光光度法测定,尿E₃、血清E₃、血清甲状腺激素[游离三碘甲状腺原氨酸(FT3)、游离甲状腺素(FT4)、促甲状腺素(TSH)]和甲状腺抗体[甲状腺球蛋白抗体(TgAb)、甲状腺过氧化物酶抗体(TpoAb)],采用酶联免疫吸附实验[ELISA]测定。在接种肿瘤后的第4日起,每3 d测量肿瘤大小。结果①各组裸鼠血清E₃和尿E₃水平比较差异有统计学意义;其中FTC+高E₃组和ATC+高E₃组的血清E(310.22、14.18 pmol/L)和尿E(36.40、8.18 pmol/L)水平均分别显著低于FTC组和ATC组的血清E(326.52、22.80 pmol/L)和尿E(38.91、11.49 pmol/L)水平;而PTC+高E₃组仅血清E₃水平(12.17 pmol/L)显著低于PTC组(23.05 pmol/L);高E₃组的尿E₃水平显著低于低E₃组(6.92、8.94 pmol/L);低E₃组的尿E₃水平显著高于对照组(8.94、7.17 pmol/L),PTC组、FTC组和ATC组的血清E₃水平显著高于对照组和高E₃组(23.05、26.52、22.80比13.23、14.46 pmol/L)。②各组血清FT3水平比较差异没有统计学意义。各组血清FT4和TSH水平与对照组比较,差异均有统计学意义,其中各实验组的FT4水平均显著低于对照组,TSH水平均显著高于对照组,但PTC组与PTC组+高E₃组、FTC组与FTC+高E₃组、ATC组与ATC+高E₃组分别进行两两比较,FT4和TSH水平并差异无统计学意义。③各组裸鼠血清TgAb和TpoAb水平比较差异均有统计学意义。其中低E₃组、高E₃组、PTC+高E₃组、FTC组、ATC组和ATC+高E₃组的TgAb水平均显著高于对照组;各实验组的TpoAb水平均显著高于对照组。④各组裸鼠尿碘水平比较差异没有统计学意义。⑤各组裸鼠肿瘤体积大小比较差异均有统计学意义,PTC+高E₃组与PTC组比较,在接种肿瘤后的第7 d、16 d、25 d、28 d和37 d,肿瘤体积发生显著性差异(71.54、226.05、730.26、893.34、1242.12比49.87、325.50、551.28、664.38、967.78 mm³);FTC+高E₃组与FTC组比较,在接种肿瘤后的第25 d、28 d、37 d和40 d,肿瘤体积发生显著性差异(392.60、475.70、794.68、943.51比214.82、238.32、422.19、490.00 mm³);ATC+高E₃组与ATC组比较,在接种肿瘤后的第37 d和40 d,肿瘤体积发生显著性差异(217.53、432.10比577.19、1146.66 mm³)。结论外源性雌三醇能够诱导裸鼠发生甲状腺功能减退和抗体水平增高,同时对乳头状甲状腺癌和滤泡状甲状腺癌的生长有着促进作用,对未分化甲状腺癌的生长有抑制作用。