广西卵形鲳鲹小脑来源细胞系的建立及特征分析

【目的】卵形鲳鲹(Trachinotus ovatus)是广西重要的海水经济鱼类,但近年来随着养殖规模扩大,各种病害侵染引起的疾病频繁暴发,给广西卵形鲳鲹养殖产业造成重大损失。建立卵形鲳鲹小脑来源的细胞系(Cell line from cerebellum of Trachinotus ovatus,TOCC),将有利于广西卵形鲳鲹养殖中病毒性病害的分离、鉴定和病害的侵染致病分子机理的研究,以及环境污染和变化的动态监测。【方法】采用胰蛋白酶消化法分离得到卵形鲳鲹小脑组织的原代细胞,并进行原代培养。待原代细胞铺满单层后进行传代培养。对稳定传代的TOCC进行18SrDNA基因测序分析,然后进行生长速率分析和染色体分析,并开展病原菌(溶藻弧菌Vibrio alginolyticus)胞外产物对TOCC的毒性分析。【结果】稳定传代的TOCC为成纤维样细胞,18SrDNA基因分析结果显示其来源于卵形鲳鲹;染色体核型分析发现TOCC具有二倍体特征,特征染色体数目为62条;不同培养条件下TOCC细胞的生长速率不同,28℃生长速度最快;病原菌胞外产物对TOCC具有明显的毒性。【结论】本研究成功建立广西卵形鲳鲹小脑组织来源的细胞系(TOCC)。细菌胞外产物对TOCC细胞毒性结果表明,该细胞系在一定范围内能够替代活体检测,实现体外细胞水平对广西海水养殖中病害侵染和环境污染物的快速检测,为养殖病害的有效预防提供技术支持。     

PHBV/葡聚糖纳米药物载体的制备及表征

两亲性聚合物纳米颗粒作为疏水性抗肿瘤药物载体因其能够增强化疗效率并降低毒副作用而受到广泛关注.采用双乳液溶剂挥发法制备了聚(3-羟基丁酸酯-co-3-羟基戊酸酯)(PHBV)/葡聚糖纳米颗粒,测得平均粒径为205.0±6.9nm,Zeta电势为-1.59±0.12mV,纳米颗粒具有明显的壳核结构,粒径均一,分散性良好.将疏水性化疗药物顺铂包载后,其粒径及电势均无明显变化,载药量达19.3±2.9%.顺铂在模拟肿瘤细胞环境pH=5.5的磷酸盐缓冲液(PBS)中比正常细胞环境pH=7.4时释放更快,且累计释放周期均长达7d以上,表明该药物载体具有一定的pH响应性以及优异的缓释性能.细胞集落形成实验表明PHBV/葡聚糖纳米药物载体具有良好的生物相容性,而载药纳米颗粒对肿瘤细胞的毒性明显高于正常细胞,表明该纳米颗粒对肿瘤细胞具有更强的杀伤作用.综上所述,PHBV/葡聚糖纳米颗粒具有两亲性分子结构,合适的粒径及Zeta电势,显著的缓释效果,对肿瘤细胞具有pH响应性及更强的杀伤作用等优势,有望成为一种新型纳米药物载体,在癌症化疗中显著提高药物利用率并降低毒副作用.     

Visual characterization of targeted effect of holo-transferrin-tagged dihydroartemisinin on human breast cancer cells

Targeted drugs could significantly reduce cytotoxic effect and increase therapeutic activity. Dihydroartemisinin (DHA) has been shown to be effective in killing cancer cells. However, it exhibits non-targeted property. Holo-transferrin (TF) is a suitable drug-carrier to target cancer cells, because cancer cells need iron uptake by the TF-mediated mechanism to maintain their uncon- trolled growth. Furthermore, TF receptors (TF-R) are highly expressed on cancer cell surfaces. In this paper, 3-(4,5-dimethylthi- azol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to evaluate the different killing effect of 4-(12-Dihydroart- emisininoxy) Benzoic Acid Hydrozide-transferrin (DBAH-TF) on human breast cancer cells (MCF-7) cells and human normal breast (HNB) cells, and atomic force microscopy (AFM) was used to visually observe the targeted effect of DBAH-TF on MCF-7 cells. MTT results show that DBAH-TF is 172 times more potent than DHA in killing MCF-7 cells, while the cytotoxic effect of DBAH-TF on HNB cells is merely 1/33 to DHA. Also, the killing effect of DBAH-TF on MCF-7 cells is 286 times that on HNB cells, showing targeted effect. Moreover, there are distinct differences in ultrastructures of cellular surfaces after DBAH-TF and DHA treatment. Through AFM imaging, many characteristic holes were observed on the cancer cell surface after being effected by DBAH-TF, which differ from the holes with irregular shapes affected by DHA. These results visually show that the DBAH-TF targeted drug has more potent killing effect on MCF-7 cells compared with DHA.     

上前牙固定修复中硅橡胶印模材料龈沟残留的观察

目的分析评估用硅橡胶二次印模法制取上前牙印模中的龈沟残留材料情况,为临床安全使用硅橡胶印模材料提供指导。方法对135颗拟行固定修复的上前基牙使用硅橡胶二次印模法制取印模后,观察并记录印模基牙龈沟翼缺损数,并依次用以下两种方法检查口内基牙龈沟中硅橡胶印模材料残留情况：A：采用肉眼直视下观察基牙龈沟,B：在5倍放大镜下继续用刮治器探寻基牙龈沟;分别记录材料残留数量与位置。结果135颗上前牙印模在肉眼直视下见24颗印模翼缘缺损,口内直视观察发现9颗基牙龈沟硅橡胶残留,继续放大镜下用刮治器探查,又发现13例龈沟深部残留。结论硅橡胶印模龈沟残留现象比较常见且容易被忽略,应引起临床医师的的高度重视,印模后应常规使用刮治器械探查龈沟内是否有印模材料的残留现象。     

丛柳珊瑚Euplexaura rhipidalis中细胞毒活性二萜成分分析

为了寻找结构新颖的海洋活性成分,对一种采自中国西沙海域丛柳珊瑚Euplexaura rhipidalis进行分离和纯化,并对获得的西松烷二菇化合物的抗肿瘤活性进行筛选,以探明该种丛柳珊瑚的活性组成成分.先利用正反相硅胶柱层析、Sephadex LH-20凝胶柱层析和半制备HPLC等方法进行分离和纯化,再利用NMR、MS等波谱方法,并与文献对照确定了3个高氧化度的西松烷二萜类化合物,分别为eorulloidolide A（1）,（1R,3S,4S,7E,11E）-3,4-epoxycembra-7,11,15-triene（2）,（-）-corulloidolide E（3）;细胞毒活性模型评价结果发现,化合物3表现了较强的细胞毒活性,对5种癌细胞肝癌HepG2,Hep3B,乳腺癌阿霉素耐药株MCF-7/ADR,前列腺癌PC-3he,结肠癌HT-116的IC50值都低于10 μg/mL.表明西松烷二萜类化合物是该丛柳珊瑚E.rhipidalis的主要活性化合物类型之一.     

Sylvaticin: A new cytotoxic and insecticidal acetogenin fromRollinia sylvatica (Annonaceae)

Summary Sylvaticin (I), a new tetrahydroxy annonaceous acetogenin with nonadjacent tetrahydrofuran rings, has been isolated from the dried fruits ofRollinia sylvatica St. Hil. (Annonaceae). This compound is extremely cytotoxic to human tumor cells and shows promising insect control properties.     

东方蝼蛄提取物对3株人宫颈癌细胞的细胞毒性研究

采用乙醇冷浸法对东方蝼蛄的干燥品粉碎物进行提取,采用聚酰胺柱层析分离划段得到东方蝼蛄乙醇提取物的不同部位,所得部位通过四甲基偶氮唑盐微量酶反应比色法(MTT法)对3种人宫颈癌细胞株(He La、Caski、C-33A)进行了体外肿瘤细胞毒性测试.结果表明,东方蝼蛄提取物中有1个分离样品的半数抑制浓度(IC50)接近10.0μg/mL,对3种人宫颈癌细胞株均具有明显的细胞毒性.东方蝼蛄乙醇提取物中存在抑制肿瘤细胞生长的物质,值得对其进一步深入研究.     

纳米银对体外培养细胞附着形态及膜功能的影响

通过研究纳米银的急性细胞毒性及其对体外培养的人脐静脉内皮细胞(HUVEC)和人脐动脉平滑肌细胞(HUASMC)附着形态、膜流动性和膜完整性的影响,初步探讨了纳米银的生物安全性.实验结果表明:浓度为0.003 9~0.5mg/mL的纳米银没有急性细胞毒性,但会影响细胞的附着形态,使细胞变小变圆,附着性变差;纳米银聚集沉积在细胞膜周围,影响细胞膜的流动性和细胞膜的完整性,从而产生一定的细胞毒性.     

羊脆木的化学成分研究(Ⅱ)

对滇产羊脆木(Pittosporum kerrii Craib)枝叶粗提物的有效部位进行化学成分研究.采用萃取、硅胶柱色谱、重结晶等方法,分离纯化得到8个单体,通过理化数据测定及波谱数据分析鉴定了它们的结构,分别是2-二十九烷酮(2-nonacosanone,1),1-二十九烷醇(nonacosan-1-ol,2),1-二十六烷醇(ceryl alcohol,3),三十二烷酸(lacceroic acid,4),三十烷酸(melissic acid,5),二十八烷酸(octacosanic acid,6),棕榈酸(palmitic acid,7),(2S,3S,4R,8E) -2-[(2R)-2-羟基二十三酰胺]-8-十八碳烯-1,3,4-三醇{(2S,3S,4R,8E)-2-[(2R)-2-hydroxytricosanoylamino]-8-octadecene-1,3,4-triol,8}.以上化合物均为首次从该植物中分离得到,化合物8为首次从该属植物中分离得到.药理试验表明化合物7、8具有细胞毒活性,对K562细胞的IC50分别为10.0、15.0 μg/mL.     

Biochemical effects and growth inhibition in MCF-7 cells caused by novel sulphonamido oxa-polyamine derivatives

The novel polyamine derivatives sulphonamido oxa-spermine (oxa-Spm) and sulphonamido oxa-spermidine (oxa-Spd) exhibited rapid cytotoxic action towards MCF-7 human breast cancer cells with IC₅₀ values of 4.35 and 6.47 μM, respectively, after 24-h drug exposure. Neither compound is a substrate of serum amine oxidase. Both oxa-Spm and oxa-Spd caused cell shrinkage, as determined by phase-contrast microscopy. After incubation with 10 μM of either compound for 8 h, the cells underwent chromatin condensation and nuclear fragmentation. However, no clear DNA ladder was obtained by electrophoresis. The sulphonamido oxa-polyamine derivatives and especially oxa-Spd enhanced the activity of polyamine oxidase (PAO), an enzyme capable of oxidising N¹-acetylated spermine and spermidine to spermidine and putrescine, respectively, generating cytotoxic H₂O₂ and 3-acetamidopropanal as by-products. The intracellular polyamine content was only marginally reduced in response to drug treatment. In conclusion, our data show that these novel sulphonamido oxa-polyamine derivatives possess high cytotoxic activity against MCF-7 cells and indicate that induction of PAO may mediate their cytotoxicity via apoptosis. Received 17 January 2002; received after revision 22 February 2002; accepted 22 February 2002