苦参碱对荷瘤小鼠抑瘤作用的实验研究

目的观察苦参碱的体内抗肿瘤活性,初步探讨其体内抑瘤作用机制。方法采用小鼠H22肝癌细胞实体瘤模型进行体内抑瘤实验,观察荷瘤动物的生长情况和肿瘤生长曲线,测定肿瘤生长抑制率;肿瘤组织病理学切片进行HE染色形态学观察,其超微结构行透射电镜观察。结果苦参碱对小鼠的H22实体瘤生长具有明显的抑制作用,抑瘤率达60％以上。苦参碱处理组小鼠的成瘤时间明显晚于生理盐水对照组,肿瘤生长也较对照组慢。组织学观察显示,肿瘤细胞大量变性坏死,电镜下可见凋亡细胞和凋亡小体。结论苦参碱具有较强的体内抗肿瘤作用,作用机制可能与其能够抑制肿瘤细胞的分裂和增殖、直接杀伤肿瘤细胞和(或)诱导肿瘤细胞凋亡、调节机体的抗肿瘤免疫应答效应有关。     

博安霉素对人食管癌等抑瘤作用的实验研究

目的：研究博安霉素（boanmycin)在体内外对人食管癌细胞等的抑瘤作用。方法：采用MTT法检测博安霉素在人体外对5种人癌细胞的细胞毒作用。同时用裸鼠异种移植人食管癌细胞模型观察博安霉素对人食管癌的抑瘤作用。结果：博安霉素在体外对5种肿瘤细胞均有明显的细胞毒作用。博安霉素在10．0mg/kg和15．0mg/kg剂量下对裸鼠异种移植人食管癌HEC2的抑制率分别为85．2％，90．6％；在7．5mg/kg和15．0mg/kg剂量下对裸鼠异种移植人食管癌Eca-109的抑制率分别为81．2％，92．0％，结论：博安霉素在体内外对人食管癌细胞等具有显著的抑瘤作用。     

龙神注射液抑瘤和急性毒性实验研究

目的了解抗癌中药龙神注射液的抑瘤作用和毒性反应。方法在小白鼠体内进行龙神注射液抑制艾氏腹水瘤作用研究和急性毒性试验。结果龙神注射液试验组小鼠平均生存天数为（２０．１７±３．６）ｄ,与生理盐水对照组相比,ｔ＝６．３５２４,Ｐ＜０．０１;ＬＤ５０＝３５６．５３ｍｇ／ｋｇ。结论龙神注射液对小鼠艾氏腹水瘤有明显抑制作用,临床使用较为安全     

狼毒蛋煎剂毒性及抑瘤效应的实验研究

目的 :研究狼毒蛋煎剂的毒性和对肉瘤S180 、HepA肿瘤细胞的抑瘤作用。方法 :①半数致死量(LD50 )毒性实验 ;②抑制HepA肿瘤细胞实验 ;③抑制肉瘤S180 实验。结果 :毒性实验 :口服狼毒蛋煎剂组LD50 >12 0 ,腹腔注射狼毒蛋煎剂组LD50 为 4 9113;口服狼毒煎剂组LD50 >12 0 ,腹腔注射狼毒煎剂组LD50 为 1 86 6 8。抑制HepA肿瘤实验 :狼毒蛋煎剂组瘤重 (0 714± 0 149)g ,5 FU组平均瘤重(0 52 1± 0 114)g ,生理盐水组平均瘤重 (9 82± 0 2 13)g。狼毒煎剂组与生理盐水组相比P <0 .0 5。抑制肉瘤S180 实验狼毒蛋煎剂组平均瘤重 1 17g ,生理盐水组平均重 2 4 6g ,两组比较P <0 .0 1。结论 :狼毒蛋煎剂能安全有效地抑制HepA肿瘤细胞和肉瘤S180 。     

宫颈癌U14细胞正丁醇提取物脂质体瘤苗的体内抑瘤效应

目的观察小鼠宫颈癌U14细胞抗原脂质体瘤苗的体内抑瘤效应。方法正丁醇法提取U14细胞表面抗原（CBE）,包裹入阴离子脂质体制成CBE-脂质体瘤苗,腹腔接种免疫Km小鼠,然后每只腹腔接种104 U14细胞或105 U14细胞,观察瘤苗对后续U14细胞负荷攻击的免疫保护作用。同时用104 U14细胞预先攻击Km小鼠,再用CBE脂质体瘤苗免疫观察瘤苗的抑瘤能力。结果低剂量（104 U14细胞）CBE脂质体瘤苗免疫组荷瘤小鼠平均生存时间为（25.50±2.08）天,明显长于空白对照组[（18.00±0.00）天]、单纯脂质体组[（18.33±0.52）天]和单纯CBE组[（19.33±1.75）天],有统计学差异（P〈0.01）。高剂量（105 U14细胞）荷瘤小鼠组与对照组生存时间比较无差异。腹腔给予104 U14细胞,再给予瘤苗免疫后各组小鼠生存时间无差异。结论 CBE脂质体瘤苗对小鼠后续腹腔攻击的相关肿瘤细胞,可产生抑制作用,但效果与肿瘤细胞负荷量有关。     

灵芝孢子粉与环磷酰胺联合使用抑瘤作用的实验研究

目的　探讨灵芝孢子粉与环磷酰胺合用对小鼠移植性动物肿瘤的抑制作用。方法　按药物体内抑瘤作用常规筛选方法进行,试验动物用NIH小鼠。瘤株为小鼠肝癌 (Hepatoma)。小鼠随机分成 4组,每组 10只。阴性对照组以 0. 5%羧甲基纤维素钠 20ml·kg-1 /d灌胃。第一实验组以环磷酰胺(CTX)20mg·kg-1 /d灌胃。第二实验组:灵芝孢子粉 4g·kg-1 /d灌胃。第三实验组:灵芝孢子粉 4g·kg-1 /d+环磷酰胺20mg·kg-1 /d灌胃,各组均于接种后 24小时开始灌胃给药,连续 10天。均于末次给药后 24小时处死,计算抑瘤率。结果　各组抑瘤率分别是:第一实验组 (CTX)为 63. 5% ~66. 1%,第二实验组为 63. 0% ~64.7%,第三实验组为 77. 8% ~79. 1%。各实验组的抑瘤作用与阴性对照组比较,差异均有非常显著性意义(P<0. 001)。孢子粉与CTX联合用药组与单用灵芝孢子粉组或单用CTX组比较,前者的抑瘤率均明显高于后两者单独用药的抑瘤率(P<0. 05)。结论　灵芝孢子粉与CTX两者联合使用具有协同抑瘤作用,灵芝孢子粉可减轻CTX的副作用。     

中药复方消核丸体外抑瘤作用的实验研究

目的研究中药复方消核丸对人乳腺癌细胞株MCF-7、人肺低分化腺癌细胞株GLC、人口腔上皮癌细胞株KB增殖的抑制作用。方法将不同浓度复方消核丸工作液分别加入含肿瘤细胞MCF-7、GLC、KB的细胞培养板中,用体外试验MTT法测定其对3种肿瘤细胞增殖的影响,试验重复3次。结果复方消核丸对MCF-7、GLC、KB肿瘤细胞的增殖有明显抑制作用（P〈0.05;P〈0.01）,且随浓度的增加,抑制率增高,呈明显剂量依赖关系,抑瘤作用的IC50分别为0.0981 g/mL、0.1149 g/mL、0.1308 g/mL。结论体外试验结果表明,复方消核丸对MCF-7、GLC、KB肿瘤细胞增殖有明显抑制作用,呈剂量依赖关系。     

红花水提物不同有效部位的体外 抗辐射作用及 机制研究

目的：通过对红花水提物中不同有效部位进行抗辐射效应及作用机制的初步探讨,为开发新型安全有效的辐射防护药物提供实验依据。方法：红花药材用蒸馏水60 ℃提取得红花水提物,采用大孔树脂HP-20洗脱分离得到水部位和30%、50%、70%、95%醇部位,采用电子自旋共振波谱仪（ESR）检测红花水提物不同有效部位清除自由基的能力;经红花水提物30%及5%醇部位处理后,利用CCK-8法检测其对⁶⁰Co γ射线照射后人肝L02细胞存活率的影响;流式细胞术研究其对⁶⁰Co γ射线照射后人淋巴细胞AHH-1细胞凋亡、细胞周期及Bcl-2/Bax比率的影响。结果：ESR自由基清除实验显示红花水提物的不同有效部位均具有清除自由基能力,半数抑制浓度IC₅₀值为1.7~79.7 mg/mL。CCK-8法检测结果显示30%及50%醇部位处理辐照后L02细胞的存活率分别为128.3%及130.9%,与照射组比较显著增加（P < 0.01）。流式细胞术结果显示与照射组凋亡率（24.7%±4.4%）比较,红花水提物30%及50%醇部位处理组凋亡率分别为15.3%±1.6%及15.2%±1.9%,明显降低（P < 0.01）。细胞周期实验结果显示与照射组AHH-1细胞G2/M期比例（45.7%±0.7%）比较,红花水提物30%及50%醇部位处理组G2/M期细胞比例分别为（42.3%±0.6%）及（36.4%±0.4%）,差异有统计学意义（P < 0.01）,同时红花水提物30%及50%醇部位能缓解照射引起的Bcl-2/Bax比率下降（P < 0.05）。结论：红花水提物的30%及50%醇洗脱部位对⁶⁰Co γ射线照射的L02、AHH-1细胞具有明显的辐射防护作用,其作用机制可能与清除自由基、改善细胞周期G2/M期阻滞、调控Bcl-2/Bax比率进而抑制细胞凋亡有关,可作为潜在的安全有效的抗辐射药物继续研究。     

PTS(抑瘤仙)体外抗瘤作用研究

目的　研究抗瘤新药PTS(抑瘤仙 )体外对肺癌细胞、耐药肺癌细胞及对正常支气管上皮细胞的作用 ,为其临床应用提供理论依据。方法　以大细胞肺癌株NCI H 460、耐药大细胞肺癌株H 460 /cDDP及正常支气管上皮细胞株 16HBE为实验对象 ,采用 2 4h活细胞跟踪法、MTT法及台盼蓝拒染记录细胞生长曲线法 ,进行PTS的体外抗瘤作用研究 ,同时以DDP(顺铂 )为阳性对照 ,PBS(平衡盐液 )为阴性对照。结果　PTS对受到直接损伤的肺癌细胞及耐药肺癌细胞均有杀伤作用 ,具有抗瘤作用和一定的抗耐药性。与DDP比较 ,PTS有更强的抗瘤作用 ,对于H 460细胞在第 48h 5 0 0mmol的DDP杀伤率为 62 .1% ,1/2 0 0的PTS为 81.3 % ;而PTS对于正常人支气管上皮 16HBE细胞的影响较小 (DDP为 3 9.5 % ,PTS为 2 1.6% ) ,具有较高的选择性。PTS应用后立即起效并达高峰 ,作用持续时间短。结论　PTS体外抗瘤作用明显 ,并具有选择性和一定的抗耐药性     

蜂胶超临界CO2萃取物的体外抗肿瘤试验研究

目的：研究蜂胶的超临界CO2萃取物的体外抗肿瘤作用。方法：以蜂胶的超临界CO2萃取物（SE-P）为原料，采用MTT法研究其对U937、95D、SGC-7901和TE-1共4株肿瘤细胞的体外抑制作用，并与市售蜂胶乙醇提取物（ME-P）和蜂胶超临界CO2萃余物95％乙醇提取物（RE-P）的抗肿瘤活性进行对比。结果：SE-P在体外对肿瘤细胞U937、95D、SGC-7901和TE-1具有较强的抑制作用，IC50分别为117．42μg／mL、138．92μg／mL、37．76μg／mL和67．89μg／mL。除95D细胞外，SE—P对其他3株肿瘤细胞的最高抑制率均高于ME-P和RE-P。结论：蜂胶的超临界CO2萃取物（SE-P）对体外培养的肿瘤细胞有较强的生长抑制作用。     

木鳖子提取物体外抗肿瘤活性的初步研究

目的：观察木鳖子（cochinchina momordica seed,CMS）水提物（water extract of CMS,CMSWE）和醇提物（ethanol extractof CMS,CMSEE）对肿瘤细胞生长的影响,探讨CMSEE的抗肿瘤机制。方法：制备CMSWE和CMSEE。MTT法检测CMSWE和CMSEE对人黑色素瘤B16、肺癌细胞A549、乳腺癌细胞MDA-MB-231、食管癌细胞TE-13,以及正常人外周血淋巴细胞（PBMC）生长的影响,计算细胞生长抑制率;光镜下观察细胞形态变化;瑞氏-吉姆萨染色观察B16细胞凋亡形态学变化。流式细胞技术（FCM）检测B16细胞凋亡率和细胞周期分布。结果：CMSEE（25—300mg／L）对B16、A549、MDA-MB231和TE-13细胞增殖均有明显的抑制作用（P〈0．01）,对PBMC的增殖没有明显影响。CMSWE（25～300mg／L）对A549、TE-13、MDA-MB231、PBMC的生长没有明显影响,仅对B16细胞的增殖有较弱的抑制作用,且各浓度组抑制率均小于相同浓度的CMSEE（P〈0．01）。经CMSEE处理后,B16细胞显示典型的凋亡形态变化,凋亡率明显增高（P〈0．01）,G0／G1期细胞明显增加,S期、G／M期细胞减少（P〈0．01）。结论：CMSEE体外对人肿瘤细胞增殖有明显的抑制作用,其抗肿瘤机制可能与阻滞肿瘤细胞周期和诱导凋亡有关。     

Anti-tumor activity of peripheral lymphocytes of tumor-bearing rats by in vitro culture

This basic study was performed to determine whether the anti-tumor effect of lymphocytes obtained from the peripheral blood of a tumor bearing host can be increased when cultured in vitro with Mitomycin C (MMC)-treated AH109A tumor cells, using interleukin-2 (IL-2). Donryu rats were used as tumor bearing hosts. The following results were obtained. Weak anti-tumor activity of lymphocytes was noted 7 days after lymphocytes were cultured in the medium to which only IL-2 was added. Anti-tumor activity was augmented by adding antigen (MMC treated tumor cells) to the above (1). Anti-tumor activity was further augmented by adding to (2) above antigen presenting cells such as intraperitoneal exudate macrophages or peripheral hole leukocytes. Anti-tumor activity of lymphocytes was detected when IL-2 and MMC treated antigen were added to the peripheral hole leukocytes containing the lymphocytes.     

In vitro anti-tumor activity of Forsythiasuspensa extract

目的:观察连翘根、叶和果3个部位各自的醇提物和水提物对食管癌细胞体外增殖和凋亡的影响,探讨连翘根醇提物(ethanol extract of Forsythia suspensa root,FSEER)的抗肿瘤机制.方法:分别制备连翘根、叶和果3个部位的醇提物和水提物,用MTF法检测6种连翘提取物对食管癌细胞TE-1、TE-13、Yes-2、Eca-109的增殖抑制作用;然后用不同浓度FSEER处理食管癌细胞24 h后用倒置相差显微镜和瑞氏-吉姆萨染色观察细胞的形态学变化,再分别用不同浓度FSEER处理TE-13细胞不同时间后,以Annexin V/PI双染法通过流式细胞仪检测TE-13细胞凋亡率的变化,Western blot检测PARP蛋白表达变化.结果:6种连翘提取物对食管癌TE-13细胞均有增殖抑制作用(P＜0.05),其中FSEER效果最明显.经FSEER处理后,TE-13细胞显示典型的凋亡形态变化,与对照组相比凋亡率明显增高(P＜0.05),PARP蛋白出现了裂解片段的表达,且呈现一定的剂量和时间依赖性.结论:FSEER体外对人食管癌细胞的增殖有明显的抑制作用,其抗肿瘤机制可能与诱导细胞凋亡有关.     

连翘提取物体外抗肿瘤活性的初步研究

目的:观察连翘根、叶和果3个部位各自的醇提物和水提物对食管癌细胞体外增殖和凋亡的影响,探讨连翘根醇提物(ethanol extract of Forsythia suspensa root,FSEER)的抗肿瘤机制。方法:分别制备连翘根、叶和果3个部位的醇提物和水提物,用MTT法检测6种连翘提取物对食管癌细胞TE-1、TE-13、Yes-2、Eca-109的增殖抑制作用;然后用不同浓度FSEER处理食管癌细胞24 h后用倒置相差显微镜和瑞氏-吉姆萨染色观察细胞的形态学变化,再分别用不同浓度FSEER处理TE-13细胞不同时间后,以Annexin V/PI双染法通过流式细胞仪检测TE-13细胞凋亡率的变化,Western blot检测PARP蛋白表达变化。结果:6种连翘提取物对食管癌TE-13细胞均有增殖抑制作用(P0.05),其中FSEER效果最明显。经FSEER处理后,TE-13细胞显示典型的凋亡形态变化,与对照组相比凋亡率明显增高(P0.05),PARP蛋白出现了裂解片段的表达,且呈现一定的剂量和时间依赖性。结论:FSEER体外对人食管癌细胞的增殖有明显的抑制作用,其抗肿瘤机制可能与诱导细胞凋亡有关。     

In vitro antimicrobial activity of propolis dry extract

In this study the antibacterial and antifungal properties of propolis, a natural product of bees, have been investigated against different pathogens. Minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBCs) were determined according to NCCLS standards on 320 strains including Staphylococcus aureus, Group A beta-hemolytic streptococci, Streptococcus pneumoniae, Moraxella catarrhalis, Haemophilus influenzae, Klebsiella pneumoniae, Escherichia coli, Proteus mirabilis, Pseudomonas aeruginosa and Candida albicans. Time-kill curves were assessed for susceptible microorganisms, testing 0, 0.5, 1, 2, 4 x MIC for propolis, by counting viable bacteria after 0, 3, 6, 24 hours and viable yeasts after 0, 3, 6, 24 and 48 hours. Propolis showed good antimicrobial activity against most of the isolates, particularly S. pneumoniae, H. influenzae and M. catarrhalis, but not against Enterobacteriaceae. Time-kill curves demonstrated bacteriostatic rather than bactericidal activity of propolis, the latter being evident only at high concentrations.     

细胞因子诱导的杀伤细胞对宫颈癌细胞的体内外杀伤效应

目的：探讨细胞因子诱导的杀伤细胞（cytokineinduced killer cells,CIKs）对宫颈癌CasKi和Hela细胞的体内外杀伤效应。方法：从宫颈癌患者的外周血分离单个核细胞,以改良方法（不加IFNγ,只加IL2 和antiCD3抗体）刺激扩增得到CIKs,用流式细胞仪分析CIKs表型。ELISA法检测CIKs培养上清中IFNγ、IL2 和TNFα的分泌水平。LDH释放法检测CIKs对CasKi和HeLa细胞的杀伤作用。建立荷CasKi或Hela细胞小鼠模型,以1×106或1×107个CIKs小鼠静脉注射治疗,每周1次,连续治疗3周,检测小鼠移植瘤的体积和重量。结果：在抗CD3抗体和IL2的刺激下,宫颈癌患者外周血单个核细胞被诱导培养成CIKs,其中CD3+CD56+细胞得到大量扩增。PHA激活后,CIKs产生大量的IFNγ和TNFα,而IL2的分泌量较少。被激活的CIKs可显著地杀伤宫颈癌CasKi细胞和HeLa细胞,最大杀伤率分别达（43.2±1.8）%和（46.2±15）%,且呈剂量依赖性。体内抗肿瘤实验结果显示,CIKs能显著抑制小鼠皮下宫颈癌移植瘤的生长（P<0.01）。结论：本实验的改良方法能有效制备CIKs,该CIKs在体内外对宫颈癌CasKi 和 Hela细胞有明显的杀伤效应。     

Anti-tumor activity of dehydroxymethylepoxyquinomicin against human oral squamous cell carcinoma cell lines in vitro and in vivo

Several reports have indicated that nuclear factor-kappa B (NF-κB) is constitutively activated in a variety of cancer cells including human oral squamous carcinoma cells, and play a key role in their growth and survival. Recent studies report that NF-κB inhibitor, dehydroxymethylepoxyquinomicin (DHMEQ), inhibits proliferation and induces apoptosis in prostate cancer cell lines. However this anti-tumor effects are still unknown in end human oral squamous carcinoma cells. In the present study, we investigated the effects of DHMEQ on oral squamous carcinoma cell (OSCC) lines in vitro and in vivo. Human OSCC cell lines (HSC-3, SAS) were treated with DHMEQ and examined for cell viability by MTT assay, cell cycle distribution by flow-cytometry, apoptosis by TUNEL assay, and protein expression by western blotting, respectively. In vivo activities were also investigated in a mouse xenograft model. DHMEQ inhibited growth of two OSCC cell lines in a dose-dependent manner measured by MTT assay. A flow cytometric analysis demonstrated that treatment with DHMEQ induced accumulation in sub-G1 phase. TUNEL assay showed that DHMEQ induced DNA fragmentation. Protein expression by western blotting analysis revealed that DHMEQ induced nuclear down regulation of Survivin, cIAP-1, and cIAP-2. In nude mice, DHMEQ inhibited growth of OSCC without major toxic side effects. The present results demonstrated that administration of DHMEQ is suggested to be a novel anti-tumor approach to the treatment of OSCC.     

Anti-tumor activity of N-thiolated beta-lactam antibiotics

An ongoing strategy for cancer treatment is selective induction of apoptosis in cancer over normal cells. N-thiolated beta-lactams were found to induce DNA damage, growth arrest and apoptosis in cultured human cancer cells. However, whether these compounds have a similar effect in vivo has not been studied. We report here that treatment with the beta-lactam L-1 caused a significant inhibition of tumor growth in a breast cancer xenograft mouse model, associated with induction of DNA damage and apoptosis in vivo. These results suggest that the synthetic antibiotic N-thiolated beta-lactams hold great potential to be developed as novel anti-cancer drugs.     

Anti-tumor activity of a novel EGFR tyrosine kinase inhibitor against human NSCLC in vitro and in vivo

Epidermal growth factor receptor (EGFR) is highly expressed in many human tumors including non-small cell lung cancer (NSCLC). Treatment with epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (TKI) has led to dramatic clinical improvement in selected patients with NSCLC. However, intrinsic and acquired resistance to EGFR-TKI remains a common phenomenon. Novel EGFR-TKI, structurally different with erlotinib or gefitinib might be beneficial for patients with NSCLC. In this study, we examined the anti-tumor effect of a newly synthesized novel EGFR tyrosine kinase inhibitor N-(3-chloro-4-fluorophenyl)-N-(7-methoxy-6-(3-morpholinopropoxy) quinazolin-4-yl)-3,3-dimethylbutanamide (F90). In vitro studies in a panel of three different human NSCLC cell lines revealed that F90 inhibited cell proliferation with high potency and induced G0/G1 arrest of cell cycle and apoptosis. F90 markedly reduced phosphorylation of EGFR and inhibited activation of MAPK and Akt. Oral administration of F90 (80 mg/kg/day) to BALB/c nude mice bearing NSCLC cell lines xenografts significantly retarded tumor growth. In conclusion, F90 has potent anti-tumor activity on human lung cancer in vitro and in vivo.