一种简单可靠的A549裸小鼠原位接种模型的建立

目的：建立一种易操作、可靠的A549原位接种模型。方法：人肺腺癌细胞株A549悬浮于matrigel中，接种于裸小鼠左肺或右侧前肢皮下，观察肿瘤生长状况及化疗药卡铂的抗肿瘤作用。结果：肿瘤在胸腔中形成，并扩展至对侧肺。与皮下肿瘤模型相比，生长在肺中的肿瘤对卡铂不敏感。结论：该A549原位接种模型简单可行，可用于筛选新型抗肺癌活性化合物。     

二硝基苯酚的抗肿瘤作用

目的探讨二硝基苯酚的抗肿瘤作用。方法采用小鼠Lewis肺癌皮下移植模型和4T1乳腺癌肺转移肿瘤模型考察二硝基苯酚的抗肿瘤作用。结果在Lewis肺癌皮下移植模型中,静注二硝基苯酚对小鼠的肺肿瘤有很好的抑制作用,局部注射没有明显的抑制作用;在4T1乳腺癌肺转移肿瘤模型中,二硝基苯酚皮下注射有明显的免疫功能和抗肺转移作用。结论二硝基苯酚具有一定的抗肿瘤作用     

海藻玉壶汤对小鼠胸腺淋巴瘤生长和转移的影响

[摘要] 目的: 建立小鼠胸腺淋巴瘤移植与转移模型,观察海藻玉壶汤对小鼠胸腺淋巴瘤生长和转移的影响。方法: 每周1次、连续5周给小鼠腹腔注射N-甲基亚硝基脲诱导小鼠胸腺淋巴瘤,将诱导的胸腺淋巴瘤经裸小鼠移植成功后制备瘤单细胞悬液,接种到小鼠皮下建立小鼠胸腺淋巴瘤移植模型,经尾静脉注射建立小鼠胸腺淋巴瘤转移模型。在模型建立同时用海藻玉壶汤进行治疗,考察其对小鼠胸腺淋巴瘤生长和转移的影响,并通过小鼠外耳微循环变化和血液流变学指标评价其治疗效果。结果: 小鼠胸腺淋巴瘤移植与转移模型建立成功,具备痰凝血瘀体征。海藻玉壶汤对小鼠胸腺淋巴瘤生长和转移均有较好的阻止作用,明显改变荷瘤小鼠的微循环障碍和高凝状态。结论: 痰凝血瘀是小鼠胸腺淋巴瘤生长和转移的重要微环境,海藻玉壶汤能阻止小鼠胸腺淋巴瘤的生长和转移。     

抗抑郁剂与组胺锌在大小鼠强迫游泳试验中的协同抗抑郁效应

目的：研究单独给予组胺锌、组胺锌伍用抗抑郁剂（丙咪嗪、文拉法辛和氟西汀）在大、小鼠强迫游泳试验中的抗抑郁效应。方法：采用小鼠强迫游泳试验，分别腹腔注射给予丙咪嗪，皮下注射给予文拉法辛、组胺锌，24小时内给药3次后观察以上药物对小鼠游泳不动时间的影响。接着采用大、小鼠强迫游泳试验两种模型，研究丙咪嗪、文拉法辛和氟西汀配伍不同比例组胺锌对动物不动时间的影响。     

三七总黄酮对病毒性心肌炎模型治疗作用的药效学研究

目的评价三七总黄酮对柯萨奇B3病毒感染体外、体内模型的治疗作用。方法采用柯萨奇B3型病毒感染原代培养Wistar乳鼠心肌细胞方法,建立体外病毒感染模型;Balb/c小鼠腹腔注射柯萨奇B3型病毒感染建立病毒性心肌炎体内实验动物模型。结果三七总黄酮能明显抑制体外培养心肌细胞病变;治疗组小鼠生存率明显增加,心肌酶释放活性明显降低;小鼠干扰素水平上升,病毒滴度降低;炎性浸润作用明显减轻。结论三七总黄酮对柯萨奇B3病毒所致小鼠病毒性心肌炎具有一定治疗作用。     

小鼠侵袭性肺曲霉病模型的建立

目的 建立侵袭性肺曲霉病(IPA)动物模型.方法 以环磷酰胺为免疫抑制剂,造成小鼠免疫抑制,对腹腔巨噬细胞和脾细胞进行计数;双侧鼻孔吸入烟曲霉菌孢子,不同时间点处死小鼠,进行组织培养及病理分析.结果 注射环磷酰胺后,小鼠免疫细胞数量急剧下降,腹腔巨噬细胞数及脾细胞数减少,外周血淋巴细胞百分比降低;对IPA模型组的组织培养可见烟曲霉,病理切片可见肺组织大量烟曲霉菌聚积,组织坏死,形成肺脓肿.结论 成功建立了小鼠IPA动物模型.     

达卡巴嗪明胶类磁性微囊制备工艺研究

目的研究几种制备达卡巴嗪明胶磁性微囊的工艺。方法化学共沉淀法制备Fe3O4磁性材料,选用不同囊材,采用溶液交联法、乳液交联法等制备磁性微囊。以微囊的包埋率、磁化率和释放性能为评价指标,比较磁性达卡巴秦微囊最佳制备工艺。结果确定最佳制备工艺。粒径约为0.05μm,药物包埋率可达71.3%,平均磁化率为25.5×10-5cm3.g-1,并具有良好的缓释性能。以明胶、壳聚糖为囊材效果更佳。     

新加沙参麦冬煎剂抑制肿瘤转移及其作用机制的实验研究

目的 探讨新加沙参麦冬煎剂防治肿瘤转移的机制。方法结合动物模型、免疫组化、细胞分子生物学技术，观察新加沙参麦冬煎剂对荷瘤小鼠抑瘤率、转移抑制率、生命延长率的影响，检测实验小鼠皮下移植瘤血管内等细胞生长因子（VEGF）、血管内皮细胞第Ⅷ因子（CD34）、黏附因子（CD44V6）、基质金属蛋白酶2（MMP2）、基质金属蛋白酶抑制酶（TIMP2）的表达情况。结果新加沙参麦冬煎剂治疗组的瘤重抑制率、肺转移抑制率、生命延长率分别为37.3%，58.3% 和20.8%；皮下移植瘤CD44V6 、VEGF表达、微血管密度（MVD）明显低于空白组（P＜0.01），TIMP2表达记分明显高出其他组（P＜0.01）。小鼠肺转移灶数与VEGF 、CD44V6、MVD呈线性相关，相关系数分别为0.490，0.398，0.455。结论①新加沙参麦冬煎剂对小鼠LA795高转移肺腺癌模型有较好抑制转移、抑制肿瘤生长和延长荷瘤小鼠生存时间的作用。②新加沙参麦冬煎剂可通过调控肿瘤转移过程中黏附、基质降解、血管生成相关分子的表达，多途径、多靶点防治肿瘤的转移。     

钙通道阻滞剂戊脉安对实验性和自发性肿瘤转移的抑制作用研究

血小板集聚过程依赖Ca~(2+)参与,而血小板集聚在肿瘤转移中起重要作用。戊脉安(Verapamil)能改变细胞Ca~(2+)环境,有证据表明它可减慢某些抗癌药的细胞外排、降低抗药性,具有抗癌协同作用。鉴此,作者观察戊脉安对肿瘤转移的影响,并初步探讨其作用机制。评价戊脉安抗转移作用分4方面进行:(1)雄性C57BL/6J 小鼠静注接种B16BL-6细胞,接种前2天开始腹腔内给药30～50mg/kg连续6天。接种后25天计数肺转移结节,有50～70%抑制率;(2)小鼠右前脚掌皮下接种B16BL-6细胞,接种后5天开始腹腔内给药,共给11天,在第17天切除带瘤肢,38天时观察淋巴结和肺转移情况。结果显示戊脉安不能抑制腋下淋巴结转移,但对肺转移约有25～80%抑制作用,在第17天测量带瘤脚掌厚度,也有轻度减少;(3)雄性BALB/c 小鼠静注接种C26NL-17细胞,腹腔内给药自接种前2天开始,共给6天,在23天观察到肺转移抑制约80%;(4)小鼠右前脚掌皮下接种C26NL-22细胞,接种第6天开始腹腔内给药共7天,第13天切除带瘤右前肢,在第29天观察到对肺转移50～60%抑制作用。对原位肿瘤也轻度抑制。同时,作者研究戊脉安对三种瘤细胞诱导对应动物血小板集聚的影响。取0.2ml 含血小板血浆(1～3.5×     

川穹嗪注射液对小鼠急性酒精性肝损伤的保护作用观察

目的：观察川穹嗪注射液对小鼠酒精性肝损伤的保护作用。方法利用50％的酒精建立急性肝损伤小鼠模型,分别以川穹嗪注射液按50、100和200 mg／kg小鼠腹腔注射给药,检测其对血清中天门冬氨酸氨基转移酶（ AST）、丙氨酸氨基转移酶（ ALT）;肝组织丙二醛（ MDA）、还原型谷胱甘肽（ GSH）含量的影响,并用苏木素－伊红（ HE）染色法观察肝组织病理学变化。结果急性酒精性肝损伤小鼠动物模型构建成功,川穹嗪注射液能够可明显降低酒精致肝损伤小鼠血清ALT和AST水平（P＜0．05）,降低小鼠肝脏组织丙二醛（MDA）含量的升高（P＜0．05）,同时使还原型谷胱甘肽（GSH）含量升高（P＜0．05）。减轻肝脏水肿、坏死等病理组织损伤。结论川穹嗪注射液对小鼠酒精性肝损伤具有保护作用,其作用机制可能与抑制氧化活性有关。     

天然产物在黑色素瘤防治中的研究进展

恶性黑色素瘤是一种高致死性的皮肤肿瘤,发病率在全世界范围内呈上升趋势。恶性黑色素瘤严重至危及生命的原因主要是其具有较强的侵袭转移能力。出现远处转移病灶的患者,即使采用最新的治疗,仍会出现复发现象,中位生存期仅有数月。目前关于黑色素瘤的防治手段主要包括手术切除、化学疗法、免疫治疗及靶向治疗。然而,这些治疗策略会导致耐药性和严重的不良反应。近年来,越来越多的研究发现天然产物具有良好的抗黑色素瘤活性,包括抑制肿瘤生长、诱导细胞凋亡、抑制血管生成和转移、消除肿瘤干细胞等。此外,许多研究报告了天然产物和传统抗黑色素瘤药物的联合作用增强了治疗效果。本文就天然产物对黑色素瘤的预防潜力、治疗效果进行简要总结讨论。     

我国临床前药物致癌试验转基因动物模型研究进展

目的：综述我国临床前药物致癌试验转基因动物模型研究进展。方法：介绍致癌试验常用转基因动物模型的构建、利用转基因动物模型开展致癌试验的优势、国际认可及国际监管机构新药申报中的应用、国内外致癌试验相关指导原则、我国开展基于转基因动物致癌试验面临的困境以及转基因动物模型构建的最新进展。结果与结论：基于转基因动物模型的致癌试验具有诸多优势也是目前国际趋势,在我国建立临床前药物致癌试验转基因动物模型非常必要。     

Late,never or non-existent: the inaccessibility of preclinical evidence for new drugs

Background and Purpose

Animal studies establish much of the evidence used to support clinical development of new drugs. Recent studies suggest that many preclinical investigations are withheld from publication, leading to exaggerated estimates of clinical utility. We sought to estimate the volume and properties of all published animal efficacy studies for a cohort of novel drugs.

Experimental Approach

We searched biomedical databases to identify 47 novel drugs whose first trials were reported between 2000 and 2003, inclusive. Next, we searched for all published animal studies testing the same drug, regardless of publication date. We then extracted items from titles and abstracts of eligible studies.

Key Results

We identified 2462 efficacy studies, representing an average of 52 studies per drug. No published efficacy studies were available for three drugs in our sample. The volume of efficacy studies was related to how far the drug had progressed in clinical development (Spearman''s correlation coefficient = 0.66, P < 0.0001). Most (87%) accessible animal efficacy studies were reported after publication of the first trial, and for 17% of the drugs in our sample, no efficacy studies were published before the first trial report. Disease indications used in trials often did not match those modelled in efficacy studies; for 35% of indications tested in trials, we were unable to identify any published efficacy studies in models of the same indication.

Conclusions and Implications

The volume of published efficacy studies is large, although numerous gaps reflect non-publication, publication delay or non-performance of efficacy studies supporting trials.     

Glycyrrhizic acid facilitates anti-tumor immunity by attenuating Tregs and MDSCs: An immunotherapeutic approach

Melanoma is one of the most aggressive malignancies and its treatment remains challenging due to its highly metastatic property and availability of limited effective drugs. In addition, immunosuppresive tumor microenvironment (TME) has been identified as major barrier to evoke anti-tumor response in melanoma. Recent studies revealed that immunosuppressive TME is directly correlated with heightened activations of T regulatory cells (Tregs) and Myeloid derived suppressor cells (MDSCs) functions. In this study, we investigated the anti-cancer effect of a triterpenoid, glycyrrhizic acid (GA) on melanoma. Our study revealed that GA not only exhibited anti-proliferative effects on melanoma cells it significantly restricted progression of melanoma tumor. However, the therapeutic efficacy of GA in impressive regression of tumor was found to be directly correlated with induction of apoptosis and modulation of cytokines from Th2 to Th1 type. To unravel the mechanism of anti-melanoma effect of GA, it has been delineated that GA inhibits pSTAT3 to evade anti-tumor suppressive function of Tregs and MDSCs. Downregulation of FOXP3, GITR and CTLA4 in tumor-infiltrating Tregs and inhibition of Cox2, PGE2 and Arginase 1 in intra-tumoral MDSC were evidenced as some of the key events during therapeutic intervention of GA in melanoma management. Moreover, GA effectively restricted advanced stage solid tumor while used in combination with Mycobacterium indicus pranii, a known immunomodulator, which alone is reported to be ineffective to restrict advanced stage solid tumor. Thus, our findings may open up a novel insight of GA as a promising agent in cancer immunotherapy or adjuvant therapy in future.     

非髓性淋巴细胞12删除对B16黑色素瘤动物模型生存期的影响

目的利用C57BL/6小鼠制备黑色素瘤动物模型,选择合适的造模条件并观察非髓性淋巴细胞删除（NMLD）对该动物模型生存期的影响。方法黑色素细胞株B16制备成不同浓度的细胞悬液,分为1×105/ml、1×106/ml、5×106/ml和1×107/ml 4组。注射于小鼠背部,每组小鼠15只。于注射7 d后采用不同方法进行全身照射,达到NMLD。观察小鼠的出瘤时间、生存期及小鼠生活习性的改变。结果 4组小鼠出瘤率均为100%。浓度越高出瘤时间及生存期越短,生活习性改变越明显。结论选择背部为注射部位,较方便。细胞浓度为1×106/ml制备的动物模型,出瘤时间和生存时间更适于实际工作;NMLD对黑色素细胞瘤动物模型的生存期没有显著影响。     

程序性死亡因子-1（PD-1）抑制药新药Nivolumab的临床研究进展

摘 要 目的： 介绍程序性死亡因子-1(PD-1)抑制药抗癌新药Nivolumab。方法: 根据文献,对Nivolumab的作用机制以及目前获批或进入III期的几个主要适应证的临床研究结果进行综述与评价。结果: Nivolumab可通过与PD-1的结合,阻断其与关键配体PD-L1和PD-L2的相互作用,恢复T细胞的抗肿瘤活性;已经完成或正在进行中的多项临床研究结果显示Nivolumab单用或者与化疗药物或CTLA-4抑制药Ipilimumab合用,对于黑色素瘤、非小细胞肺癌以及肾细胞癌的效果优于目前临床应用的化疗药物,Nivolumab总体耐受性好,3～4级的不良事件主要为免疫介导肺炎,肝功能紊乱,疲劳等。结论:Nivolumab通过肿瘤免疫效应可改善对黑色素瘤、非小细胞肺癌以及肾细胞癌等多种肿瘤的治疗效果。     

In vitro and in vivo anti-melanoma action of metformin

The in vitro and in vivo anti-melanoma effect of antidiabetic drug metformin was investigated using B16 mouse melanoma cell line. Metformin caused a G₂/M cell cycle arrest associated with apoptotic death of melanoma cells, as confirmed by the flow cytometric analysis of cell cycle/DNA fragmentation, phosphatidylserine exposure and caspase activation. Metformin-mediated apoptosis of melanoma cells was preceded by induction of oxidative stress and mitochondrial membrane depolarization, measured by flow cytometry in cells stained with appropriate fluorescent reporter dyes. The expression of tumor suppressor protein p53 was increased, while the mRNA levels of anti-apoptotic Bcl-2 were reduced by metformin, as revealed by cell-based ELISA and real-time RT-PCR, respectively. Treatment with metformin did not stimulate expression of the cycle blocker p21, indicating that p21 was dispensable for the observed cell cycle arrest. The activation of AMP-activated protein kinase (AMPK) was not required for the anti-melanoma action of metformin, as AMPK inhibitor compound C completely failed to restore viability of metformin-treated B16 cells. Metformin induced autophagy in B16 cells, as demonstrated by flow cytometry-detected increase in intracellular acidification and immunoblot-confirmed upregulation of autophagosome-associated LC3-II. Autophagy inhibitors ammonium chloride and wortmannin partly restored the viability of metformin-treated melanoma cells. Finally, oral administration of metformin led to a significant reduction in tumor size in a B16 mouse melanoma model. These data suggest that anti-melanoma effects of metformin are mediated through p21- and AMPK-independent cell cycle arrest, apoptosis and autophagy associated with p53/Bcl-2 modulation, mitochondrial damage and oxidative stress.     

三七总皂苷对B16黑色素瘤生长及转移的作用

目的观察三七总皂苷（PNS）对B16黑色素瘤生长及转移的作用,及其对B16黑色素瘤表达缝隙连接蛋白Connexin32的影响。方法实验采用B16黑色素瘤自发性肺转移模型,来观察PNS对B16黑色素瘤生长及转移的影响。免疫组织化学检测Connexin32在黑色素瘤原发灶的表达情况。结果（1）PNS对黑色素瘤生长有较好的抑制作用,其中PNS高剂量组抑瘤率可达50．85％。（2）与模型组相比PNS中、高剂量组能有效抑制黑色素瘤的肺转移,转移灶数量与对照组相比有明显减少。（3）免疫组织化学检测,发现PNS各用药组均能增强黑色素瘤原发灶细胞膜上Connexin32的表达。结论PNS能够抑制B16黑色素瘤的生长和转移,并能有效增强肿瘤细胞膜上Connexin32的表达。