ChIL-2对H5亚型AIV疫苗免疫增强作用研究   总被引：4，自引：0，他引：4       下载免费PDF全文

刘岩  由振强  许健  郑江涛  于涟 《浙江大学学报(农业与生命科学版)》2005,31(6):797-801

鸡白细胞介素2真核表达质粒pCI-IL-2及原核表达重组鸡白细胞介素2(rChIL-2)分别与H5亚型禽流感油乳剂灭活苗联合使用,25日龄首免,40日龄时加强免疫.MTT法检测各组鸡外周血淋巴T细胞增殖,HI试验监测H5亚型禽流感抗体水平的消长规律,结果表明:pCI-IL-2及rChIL-2均能显著增强AIV疫苗的免疫效果(P＜0.05),pCI-IL-2和rChIL-2两组间差异不显著(P＞0.05);证实鸡白细胞介素2对H 5亚型禽流感疫苗有免疫增强作用.rChIL-2协同免疫组MTT及HI试验均在第四周 达到峰值,pCI-IL-2协同免役组峰值出现时间稍晚,但随后质粒免疫组可维持在更高的水平值.提示IL-2蛋白的作用更迅速,IL-2质粒作用更持久.  相似文献   

聚乙二醇化重组人白细胞介素6的放射性标记     

徐贤坤  王霜  曾宪垠  汪淑芳 《核农学报》2006,20(3):236-240

分别用常规氯氨T和改进的双相氯氨T法对PEG-rhIL-6进行放射性碘标记,用柱层析法和离心超滤法对标记物分离纯化,同时用三氯乙酸沉淀法和SDS-PAGE电泳法鉴定纯化后的标记化合物放射化学纯度,采用rhIL-6依赖细胞7TD1,用MTT比色法测定标记物的生物学活性。结果表明:1.改进的双相氯氨T法标记率为74.5%,高于常规氯氨T法的62.3%,放射性比活度分别为5.513×105和4.610×105Bq/μg。2.经柱层析法和离心超滤法对标记物分离纯化后,用三氯乙酸沉淀法测得放射化学纯度均能达到99%以上。3.用SDS-PAGE电泳法检测两种标记方法所得的标记物与非标记物的结果表明,常规氯氨T法标记物比非标记物多1条高分子量蛋白带,认为是标记过程中蛋白质受到部分损伤;改进的双相氯氨T的标记物与非标记物蛋白质条带一致,认为蛋白质标记后基本没有受到损伤。4.生物活性检测表明改进的双相氯氨T标记物与非标记物活性之间无显著差异,常规氯氨T法标记物活性略低于双相氯氨T的标记物活性。  相似文献   

MTT比色法在传染性法氏囊病病毒分离中的应用     

崔言顺  马秀丽  张秀美  黄兵 《中国兽医学报》2003,23(6):543-544

应用MTT比色法检测了病料中传染性法氏囊病病毒(IBDV)在细胞培养物中的增殖情况，并对病料中加入法氏囊提取液、鸡胚浸出液2种成分对病毒增殖的影响作了比较。结果表明，MTT比色法简便、快速、可靠，能很好地反映病毒在细胞培养物中的增殖情况。  相似文献   

金顶侧耳多糖体外抗肿瘤作用的研究   总被引：3，自引：0，他引：3  

王晓洁  蔡德华  杨立红  卜庆梅 《食用菌学报》2005,12(1):9-13

通过深层发酵获得金顶侧耳菌丝体。用水提法分别提取金顶侧耳菌丝体多糖、胞外(过滤液)多糖和全液(菌丝体 发酵液)多糖。用MTT比色法测定金顶侧耳多糖体外对小鼠S180癌细胞及人结肠低分化腺癌细胞的抑制率。结果表明,金顶侧耳胞外多糖对体外培养的S-180癌细胞有抑制作用,全液多糖和菌丝体多糖无抑制作用。这3种多糖体外对人结肠低分化腺癌细胞有抑制作用,其中金顶侧耳胞外多糖抑制率最高,全液多糖次之,菌丝体多糖最低。  相似文献   

小鼠精原干细胞分离培养条件的优化     

张秀娟  李莉  卫恒习  白银山  师如意  张守全 《华南农业大学学报》2012,33(1):87-91

采用两步酶消化法获得5～7日龄昆明小鼠睾丸组织的单细胞悬液,比较差速贴壁法和Percoll不连续密度梯度法对精原干细胞(Spermatogonial stem cells,SSCs)的纯化效果,并利用MTT法检测不同生长因子及添加浓度对SSCs体外增殖的影响.结果表明5日龄小鼠和7日龄小鼠的睾丸细胞总数存在较大差异(P<0.05),但SSCs数目无显著差异(P>0.05);与Percoll不连续密度梯度法相比,差速贴壁法获得了较高的SSCs数量和纯化率,其纯化率平均85.06%;MTT法检测结果表明LIF因子的添加对SSCs的增殖具有促进作用,20 ng/mL GDNF+20 ng/mLbFGF+103 U/mL LIF的组合添加是昆明小鼠SSCs的体外培养增殖的最佳组合.  相似文献   

长春花4种提取物肝癌细胞毒活性的研究   总被引：1，自引：0，他引：1  

曾艳波  梅文莉  董文化  李小娜  戴好富 《热带农业科学》2012,32(9):41-43

用溶剂萃取法对长春花地上部分的95%乙醇提取物进行分段处理,以肝癌细胞系SMMC-7721为筛选模型,利用MTT法测定各提取物的体外肿瘤细胞毒活性,对长春花的肝癌细胞毒活性部位进行了筛选。结果表明:长春花乙醇总提取物和正丁醇提取物对体外肝癌细胞表现出较强的抑制活性,石油醚提取物﹑乙酸乙酯提取物和水提取物则没有活性。说明正丁醇提取物部分是长春花体外肝癌细胞毒活性部位。  相似文献   

Antiproliferative activity of Saponaria vaccaria constituents and related compounds     

Balsevich JJ  Ramirez-Erosa I  Hickie RA  Dunlop DM  Bishop GG  Deibert LK 《Fitoterapia》2012,83(1):170-181

Total methanolic extracts of Saponaria vaccaria seed derived from several varieties, as well as various purified components obtained through successive chromatographic separations of total extracts were evaluated for their growth inhibitory activity in WiDr (colon), MDA-MB-231 (breast), NCI-417 (lung) and PC-3 (prostate) human cancer cells as well as the non-tumorigenic fibroblast BJ (CRL-2522) cell line using MTT colorimetric assay. Purified bisdesmosidic saponins segetoside H and I were further examined using microscopy and apoptosis assays. Bisdesmosidic saponins exhibited dose-dependent growth inhibitory and selective apoptosis-inducing activity. Growth inhibitory effects were particularly strong in a breast (MDA-MB-231) and a prostate (PC-3) cancer cell line. Total extracts exhibited a different preference being most active against a colon cancer cell line (WiDr). In a comparison of varieties, all of the total seed extracts exhibited similar dose-dependent activities, but with some variation in potency. Monodesmosidic saponins vaccarosides A and B, phenolic vaccarin, and cyclopeptide segetalin A, co-occurring seed substituents, did not exhibit activity. The non-tumorigenic fibroblast cell line BJ (CRL 2522) was growth inhibited but did not undergo apoptosis when treated with bisdesmosidic saponins at low micromolar concentrations. Saponin-rich extracts from Kochia scoparia seed and Chenopodium quinoa were also evaluated alongside Saponaria saponins but did not exhibit activity. Closely related Quillaja saponins exhibited activity but were less potent.  相似文献   

Cytotoxic, cytoprotective and antioxidant effects of isolated phenolic compounds from fresh ginger     

Peng F  Tao Q  Wu X  Dou H  Spencer S  Mang C  Xu L  Sun L  Zhao Y  Li H  Zeng S  Liu G  Hao X 《Fitoterapia》2012,83(3):568-585

Twenty-nine phenolic compounds were isolated from the root bark of fresh (Yunnan) ginger and their structures fully characterized. Selected compounds were divided into structural categories and twelve compounds subjected to in-vitro assays including DPPH radical scavenging, xanthine-oxidase inhibition, monoamine oxidase inhibition, rat-brain homogenate lipid peroxidation, and rat pheochromocytoma PC12 cell and primary liver cell viability to determine their antioxidant and cytoprotective properties. Isolated compounds were also tested against nine human tumor cell lines to characterize anticancer potency. Several diarylheptanoids and epoxidic diarylheptanoids were effective DPPH radical scavengers and moderately effective at inhibiting xanthine oxidase. An enone–dione analog of 6-shogaol (compound 2) was isolated and identified to be most effective at protecting PC12 cells from H₂O₂-induced damage. Almost all tested compounds inhibited lipid peroxidation. Three compounds, 6-shogaol, 10-gingerol and an enone-diarylheptanoid analog of curcumin (compound 6) were identified to be cytotoxic in cell lines tested, with KB and HL60 cells most susceptible to 6-shogaol and the curcumin analog with IC₅₀ < 10 μM. QSAR analysis revealed cytotoxicity was related to compound lipophilicity and chemical reactivity. In conclusion, we observed distinct compounds in fresh ginger to have biological activities relevant in diseases associated with reactive oxygen species.  相似文献   

Echinacoside promotes bone regeneration by increasing OPG/RANKL ratio in MC3T3-E1 cells     

Fei Li  Yanan Yang  Panpan Zhu  Weina Chen  Dongli Qi  Xiupu Shi  Chunfeng Zhang  Zhonglin Yang  Ping Li 《Fitoterapia》2012

Echinacoside (ECH), isolated from Cistanche tubulosa (Schrenk) R. Wight stems, was subjected to in vitro experiments to investigate its bioactivities on proliferation, differentiation and mineralization of the osteoblastic cell line MC3T3-E1. MTT assay, the alkaline phosphatase (ALP) activity and calcium deposition were determined, and the secretion of collagen I (COL I), osteocalcin (OCN), osteoprotegerin (OPG) and receptor activator of nuclear factor-κB ligand (RANKL) were also assayed by enzyme-linked immunosorbent assay (ELISA). The results showed that ECH caused a significant increase in cell proliferation, ALP activity, COL I contents, OCN levels and an enhancement of mineralization in osteoblasts at the concentration range from 0.01 to 10 nmol·L^− 1 (p < 0.05), suggesting that ECH has a stimulatory effect on osteoblastic bone formation or has potential activity against osteoporosis. In addition, the ratio of OPG/RANKL also could be enhanced by ECH. These findings provide the potent evidence that ECH can promote bone regeneration in cultured osteoblastic MC3T3-E1 cells, which might be done by elevating the OPG/RANKL ratio, and potential evidence for echinacoside to be a promising drug or a lead compound in the development of disease-modifying drug to prevent osteoporosis.  相似文献   

鹅副黏病毒与新城疫病毒对鸡胚和鹅胚成纤维细胞的感染性分析     

单松华  谢爱织  邹键  姚龙涛 《动物医学进展》2005,26(2):73-76

用鸡新城疫病毒F48E9,Komarov, LaSota,V4/66株和鹅副黏病毒SF02株分 别感染鸡胚和鹅胚成纤维细胞,用MTT显 色方法检测存活细胞数目。结果表明,不同 毒力新城疫病毒致细胞病变的作用不一样, 鸡新城疫强毒F48E9株和鹅副黏病毒SF02 株均引起两种细胞几乎全部死亡,感染动物 时,SF02与NDV强毒株的致病性有显著的 区别,但感染细胞时,二者并无明显差别; NDV与GPMV在感染水禽时的致病性差异 并不是因为诸如细胞膜受体等细胞水平的因 素不同造成的,可能与干扰素途径有关。  相似文献