反义局部粘着斑激酶抑制肝癌侵袭生长的研究

目的　观察反义局部粘着斑激酶 (FAK)脱氧寡核苷酸 (ODN)转染对Bel740 2肝癌移植瘤侵袭性生长的影响 ,并探讨其作用机制。方法　以LipofectAMINE介导的反义FAKODN转染Bel 740 2肝癌细胞株后 ,于 6只裸鼠双侧颈背部、髋部共 4处皮下接种 ,观察移植瘤生长情况 ,并行肿瘤微血管密度 (MVD)的免疫组织化学检测及MMP2与TIMP2表达的逆转录 聚合酶链反应(RT PCR)检测。结果　反义FAKODN转染使裸鼠皮下移植瘤的生长受到显著抑制 ,抑瘤率为3 5 .7% ;MVD在反义转染组 ( 9.5 99± 1.2 84)显著低于对照组 [( 13 .915± 2 .618) ,P <0 .0 5 ] ;MMP2表达在反义转染组 ( 0 .199± 0 .0 61)显著低于对照组 ( 0 .42 1± 0 .118) ,TIMP2表达在反义转染组( 0 .461± 0 .15 3 )则显著高于对照组 [( 0 .2 98± 0 .10 4) ,P <0 .0 5 ]。结论　信号转导分子FAK表达阻断可显著抑制Bel 740 2肝癌细胞株在裸鼠体内的侵袭性生长 ,病理性肿瘤血管生成减少及MMP2、TIMP2表达改变与其抗肿瘤作用密切相关。     

利拉鲁肽极化M2型巨噬细胞改善非酒精性脂肪肝的机制

目的：探讨利拉鲁肽改善2型糖尿病伴随的非酒精性脂肪肝（NAFLD）的作用机制。方法：小鼠随机分为正常组、模型组和利拉鲁肽组3组,每组15只。模型组和利拉鲁肽组采用高糖高脂饲料联合小剂量链脲佐菌素的方法,构建2型糖尿病NAFLD的小鼠模型,利拉鲁肽组皮下注射利拉鲁肽（100 μg/kg）,每日1次,连续4周。检测各组小鼠的体质量、肝指数、ALT、AST和TG指标;HE染色及油红O染色观察肝组织病理改变;ELISA法检测血清中IL-4和IL-10的含量;流式细胞术检测M2型巨噬细胞比例;RT-PCR检测IL-4、IL-10、CD206、CD301、MGL-1、MGL-2和Arg-1 mRNA表达。结果：与模型组比,利拉鲁肽组小鼠体质量、肝指数、TG、AST和ALT均明显下降,肝组织中脂肪变性减轻,M2型巨噬细胞比例显著上升（P＜0.001）,IL-4、IL-10蛋白表达显著上升（P＜0.05）,IL-4、IL-10、CD206、CD301、MGL-1、MGL-2和Arg-1 mRNA表达也显著上升（P＜0.01）。结论：利拉鲁肽对2型糖尿病NAFLD具有保护作用,其机制可能与促进IL-4和IL-10的表达,极化M2型巨噬细胞有关。     

NOD/SCID雌性小鼠动情周期的观察

目的：观察NOD/SCID小鼠动情周期及卵巢切除术后阴道涂片的变化。方法：连续观察9天NOD/SCID鼠,每日进行两次阴道涂片,计算动情周期时间及发生率。卵巢切除后,阴道涂片并观察阴道涂片变化。结果：NOD/SCID小鼠动情周期为4-6天。有规律动情的小鼠占80%。阴道口状态与动情周期无明显相关。卵巢切除术后,阴道涂片呈动情后期或动情间期改变。结论：采用阴道脱落细胞涂片法可判断NOD/SCID雌性小鼠的动情周期及特点,NOD/SCID雌性小鼠卵巢切除手术有效。     

人结肠鳞癌直肠鳞腺癌SCID鼠原位移植高转移模型的建立及其生物学特性的研究

目的 建立人结肠鳞癌直肠鳞腺癌SCID鼠原位移植高转移模型,为探讨理想的治疗结肠癌肝转移和预防结肠癌根治术后肝转移的方法提供实验工具。方法 采用组织学完整的结肠癌手术标本植入SCID鼠结直肠（粘膜层）壁内,观察原位移植的成瘤、移植瘤的侵袭和转移及其形态学特征（光镜、电镜、免疫组织化学）。结果 两株人结肠癌SCID鼠均获原位移植成功。人结肠鳞癌SCID鼠原位移植模型HCS-HMN-1已传至19代,人直肠鳞癌SCID鼠原位移植模型HRSA-HMN-2已传至23代,共移植SCID鼠223只,其移植生长率和自发转移率及液氮冻存复苏成活率均为100％,移植瘤在结直肠内呈广泛原位侵袭性生长、淋巴结转移、肝转移和全腹腔播散转移。并具有分泌CEA的功能。移植瘤细胞病理学和电镜观察、流式细胞仪DNA含量检测及染色体核型分析与瘤源人结直肠癌细胞完全一致。结论 本研究所建立的两株人结直肠癌SCID鼠高转移模型完整地模拟了人结直肠癌侵袭和转移的临床过程。为进一步研究结直肠癌肝转移、治疗和预防结直肠癌根治术后肝转移的方法提供了理想的实验动物模型。     

Introducing the clusterin gene into human renal cell carcinoma cells enhances their metastatic potential

PURPOSE: We recently reported a protective role of clusterin expression against apoptosis induced by a wide variety of stimuli in several human cancer models. In the current study we tested the hypothesis that clusterin over expression confers a benefit for the metastasis of renal cell carcinoma through the inhibition of apoptosis induced by the various obstacles the cancer cells may confront after detachment from their primary origin. MATERIALS AND METHODS: We introduced clusterin complementary DNA into human renal cell carcinoma ACHN cells, which do not express detectable level of clusterin expression, and generated the clusterin over expressing cell line ACHN/CL and the control vector only transfected cell line ACHN/C. In vitro anti-cell death activity under anchorage independent conditions among ACHN sublines was examined by limiting dilution assay and cell survival assay in suspension. To investigate the in vivo effects of clusterin over expression on metastatic potentials each cell line was injected into the tail vein or renal subcapsule of nonobese diabetic, severe combined immunodeficient mice and the metastatic features in all abdominal and thoracic organs were evaluated. RESULTS: ACHN/CL showed significantly enhanced growth in limiting dilution cultures compared with ACHN/C. The analysis of cell survival in the floating assay also revealed that ACHN/CL had a powerful survival advantage in suspension compared with ACHN/C. Furthermore, ACHN/CL formed more than 5-fold as many metastatic nodules in the lung after intravenous injection than ACHN/C. Similarly more marked lung metastasis was observed after implanting ACHN/CL cells into the renal subcapsule than after implanting ACHN/C cells. In contrast, there were no significant differences among ACHN sublines in the growth rates in vitro and in vivo, cell motility or invasive ability. CONCLUSIONS: These findings suggest that, if clusterin is over expressed, it prolongs cell survival under unfavorable conditions in the metastatic process, resulting in the enhanced metastatic potential of renal cell carcinoma.     

植入用缓释氟尿嘧啶治疗结直肠癌的实验研究

目的 研究氟尿嘧啶（5-Fu）缓释植入剂对结直肠癌移植瘤的治疗效果.方法 将50只直肠癌荷瘤鼠随机分为5组,每组10只.A、B组于瘤周植入5-FU缓释剂,剂量分别为200mg/kg和100 mg/kg;C、D组于瘤周注射5-FU注射液,剂量分别为200 mg/kg和100 mg/kg;E组不予任何治疗.分别于给药后0、3、6、9和12 d,观察裸鼠生存情况、体质量变化及肿瘤体积,12 d后处死小鼠.结果 A、B组肿瘤生长曲线平缓,12 d后A、B、C及D组抑瘤率分别为72%、51%、8%及5%,A、B组肿瘤体积与C、D组比较,差异有统计学意义（P＜0.05）.A、B、C和D组在给药3 d后体质量下降,其中以C组最为明显;以后各组体质量增加,12 d后,各组小鼠体质量间差异无统计学意义（P＞0.05）.在实验过程中,A、B、C和D组小鼠分别死亡1、0、4和1只.结论 5-FU缓释植入剂于瘤周植入能安全有效地抑制结直肠癌移植瘤的生长.     

前列腺的小细胞神经内分泌癌：异种移植是一个更好的实验模型吗？

前列腺小细胞神经内分泌癌（SCNCP）是一种罕见的前列腺癌,临床恶性程度高,预后差,对其研究在临床上具有重大意义。目前,研究SCNCP的方法有人工肿瘤细胞系培养法,还有将从患者身上直接获得的新鲜肿瘤组织切片植入免疫缺陷宿主小鼠体内的方法。本综述的目的是整合20多年在前列腺小细胞神经内分泌癌的异种移植的研究数据。异种移植提供的有关数据包括组织病理学、核型、DNA含量、细胞周期频率、肿瘤标志物、雄激素受体表达、转移和成功率。我们尽可能的比较了患者体内获得的原始原位切片和宿主小鼠异种移植组织,发现小细胞神经内分泌前列腺癌异种移植与培养细胞异源移植相比各有千秋。总体而言,肿瘤异种移植均优于传统的异源移植,主要表现在保持肿瘤形态、病理、分泌活动和病人原始样本肿瘤标记物的表达等方面。此外,异种移植组织保留了前列腺肿瘤的三维结构,以保持其关键的基质上皮细胞相互作用。     

裸鼠原位和异位脑肿瘤生物发光信号成像实验研究

目的应用生物发光成像技术,非侵入性地连续检测活体裸鼠原位和异位脑肿瘤发展演进过程。方法用SMPU-R-MND-luc载体转染人脑肿瘤U87MG细胞系,形成具有高荧光素酶活性的细胞克隆。在裸鼠脑内和胁腰部皮下植入持续表达荧光素酶的肿瘤细胞,建立原位和异位脑肿瘤模型,用影像学资料显示肿瘤部位。用光子发射定量分析动态监测肿瘤生长情况。结果成功地建立了表达荧光素酶活性的原位和异位脑肿瘤动物模型。采集反映肿瘤生长的生物发光信号,肿瘤细胞植入后不同时间点的发光信号值呈显著正相关,而且原位和异位脑肿瘤间存在明显差异。但生物发光脑肿瘤生物发光信号值在第4 d和第14 d时无显著差异。结论体内生物发光成像可以非侵入性地动态检测活体内脑肿瘤演进过程,为研究肿瘤发展机制及最佳治疗策略的选择提供了新的手段和工具。     

Progressive hearing loss in mice carrying a mutation in the p75 gene

The neurotrophin receptor p75 (p75NTR) is expressed in the developmental stage of the cochlea. However, the role of the p75NTR in the inner ear remains to be established. In this study, we conducted electrophysiological and morphological analyses of the auditory function of mice carrying a mutation in the p75 gene at different longitudinal stages. The mice carrying a mutation in the p75 gene showed an age-related progressive hearing loss. At 1 month, there was no obvious morphological change in the cochlea of the mice carrying a mutation in the p75 gene compared to wild-type mice, except for a slight loss of spiral ganglion neurons (SGNs). Auditory function was not significantly different between both genotypes from 1 to up to 4 months of age. The mice carrying a mutation in the p75 gene started to show progressive hearing loss at 4 months, when both SGN degeneration and hair cell (HC) loss were observed at the basal turn. These results suggest that the neurotrophin receptor p75 may play a significant role in the maintenance of cochlear function, and that mice carrying a mutation in the p75 gene could be a good animal model of early onset progressive hearing loss.     

Estradiol stimulates progenitor cell division in the ventricular and subventricular zones of the embryonic neocortex

Two distinct populations of cerebral cortical progenitor cells that generate neurons during embryogenesis have been identified: radial glial cells and intermediate progenitor cells. Despite advances in our understanding of progenitor cell populations, we know relatively little about factors that regulate their proliferative behaviour. 17-beta-Estradiol (E2) is present in the adult and developing mammalian brain, and plays an important role in central nervous system processes such as neuronal differentiation, survival and plasticity. E2 also stimulates neurogenesis in the adult dentate gyrus. We examined the role of E2 during embryonic cortical neurogenesis through immunohistochemistry, in situ hybridization, functional enzyme assay, organotypic culture and in utero administration of estradiol-blocking agents in mice. We show that aromatase, the E2 synthesizing enzyme, is present in the embryonic neocortex, that estrogen receptor-alpha is present in progenitor cells during cortical neurogenesis, that in vitro E2 administration rapidly promotes proliferation, and that in utero blockade of estrogen receptors decreases proliferation of embryonic cortical progenitor cells. Furthermore, the E2 inhibitor alpha-fetoprotein is expressed at high levels by radial glial cells but at lower levels by intermediate progenitor cells, suggesting that E2 differentially influences the proliferation of these cortical progenitor cell types. These findings demonstrate a new functional role for E2 as a proliferative agent during critical stages of cerebral cortex development.