MTT法检测番鸭外周血淋巴细胞转化功能的研究   总被引：8，自引：3，他引：5  

林锋强  胡奇林  陈少莺  陈仕龙  程晓霞  朱小丽 《动物医学进展》2005,26(1):75-77

为了建立番鸭淋巴细胞转化检测的MTT法 ,筛选细胞浓度、刀豆蛋白ConA浓度和培养时间 3个参数对试验条件进行了研究。结果确定了MTT法检测番鸭淋巴细胞转化能力的最佳培养条件 ,细胞浓度 5× 1 0 6个 /mL在 2 0 μg/mLConA的RMPI 1 640完全培养基 40℃培养 60h。试验表明 ,该方法可用于番鸭体外细胞免疫功能的检测  相似文献   

Development and Characterization of Calcium-Alginate Beads of Apigenin: In Vitro Antitumor,Antibacterial, and Antioxidant Activities     

Mohammed F. Aldawsari  Mohammed Muqtader Ahmed  Farhat Fatima  Md. Khalid Anwer  Prakash Katakam  Abdullah Khan 《Marine drugs》2021,19(8)

The objective of this work was to develop sustained-release Ca-alginate beads of apigenin using sodium alginate, a natural polysaccharide. Six batches were prepared by applying the ionotropic gelation technique, wherein calcium chloride was used as a crosslinking agent. The beads were evaluated for particle size, drug loading, percentage yield, and in vitro drug release. Particle size was found to decrease, and drug entrapment efficiency was enhanced with an increase in the polymer concentration. The dissolution study showed sustained drug release from the apigenin-loaded alginate beads with an increase in the polymer proportion. Based on the dissolution profiles, BD6 formulation was optimized and characterized for FTIR, DSC, XRD, and SEM, results of which indicated successful development of apigenin-loaded Ca alginate beads. MTT assay demonstrated a potential anticancer effect against the breast cancer MCF-7 cell lines. The antimicrobial activity exhibited effective inhibition in the bacterial and fungal growth rate. The DPPH measurement revealed that the formulation had substantial antioxidant activity, with EC50 value slightly lowered compared to pure apigenin. A stability study demonstrated that the BD6 was stable with similar (f2) drug release profiles in harsh condition. In conclusion, alginate-based beads could be used for sustaining the drug release of poorly water-soluble apigenin while also improving in vitro antitumor, antimicrobial, and antioxidant activity.  相似文献   

重组红火蚁毒素蛋白SoliⅣ致病机理的初步研究     

王建峰  林祥梅  张书霞  韩雪清  吴绍强  杨泽晓 《中国畜牧兽医》2008,35(1):83-86

作者通过研究重组红火蚁毒素蛋白Solenopsis invictaⅣ (SoliⅣ)对兔及外周血淋巴细胞的影响,探讨红火蚁毒素蛋白致病机制。通过分离兔外周血淋巴细胞进行培养,经不同浓度的重组红火蚁毒素蛋白SoliⅣ分别和细菌脂多糖(LPS)、刀豆蛋白(ConA)共同刺激后,MTT法测定淋巴细胞的增殖情况;体内试验：用重组蛋白SoliⅣ从兔背部皮下注入,观察兔的临床反应,定期采血,用ELISA方法测定兔血清中白介素 4（IL-4）和总IgE的变化。重组红火蚁毒素蛋白SoliⅣ浓度为25、50、75 μg/ml和LPS共刺激时,与单独LPS刺激对照组相比,淋巴细胞增殖活性显著增高（P<0.05）;浓度为15、25、50、75 μg/ml和ConA共刺激时,与单独ConA刺激对照组比较,淋巴细胞增殖活性显著增高（P<0.05）;重组SoliⅣ过敏兔的血清中IL-4和总IgE的水平升高,在20 h左右达到最高值。试验结果表明,一定浓度的重组红火蚁毒素蛋白SoliⅣ能引起体外培养的T、B淋巴细胞增殖,过敏体质兔在重组红火蚁毒素蛋白SoliⅣ刺激后能引起Ⅰ型变态反应。  相似文献   

猪外周血单个核细胞增殖反应影响条件的探讨   总被引：8，自引：0，他引：8  

宋长绪  谢明权 《中国预防兽医学报》1999,21(4):289-290

应用 Ｌ１６ （４５） 正交试验, 对影响猪 Ｐ Ｂ Ｍ Ｃ 增殖反应的几个因素, 包括培养时间、 Ｐ Ｈ Ａ 浓度、细胞浓度等三个因素四个水平进行了比较和探索。试验表明几个因素对 Ｐ Ｂ Ｍ Ｃ 的增殖反应均有显著的影响（ Ｐ＜ ００５） , Ｐ Ｂ Ｍ Ｃ 增殖反应的 Ｍ Ｔ Ｔ 比色法的最佳反应条件组合为 Ｐ Ｂ Ｍ Ｃ 浓度为１ ×１０６／ｍｌ, 植物血凝素 Ｐ Ｈ Ａ 的浓度为１２５μｇ／ ｍｌ, 细胞培养时间为２４ 小时; 影响增殖反应的先后顺序为 Ｐ Ｂ Ｍ Ｃ 浓度、 Ｐ Ｈ Ａｐ 浓度及细胞培养时间。同时确定了最有利于 Ｐ Ｂ Ｍ Ｃ 增殖的犊牛血清浓度为１０ ％ 。  相似文献   

用于水生病原菌的3种高通量活菌计数方法的比较     

邹培卓  杨倩  董宣  谢国驷  黄捷 《渔业科学进展》2019,40(3):133-140

以一株副溶血弧菌(Vibrio parahaemolyticus)作为研究对象,比较了MTT [3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐]比色法、ATP生物发光法和高通量生长曲线法在活细菌高通量计数上的应用效果。用96孔培养板进行不同浓度细菌活菌计数,确定了上述3种方法在副溶血弧菌活菌计数的标准曲线和线性范围。结果显示,副溶血弧菌的MTT比色法以DMSO溶解的MTT产物甲瓒在555 nm的吸光度(OD555 nm)为计数依据,活菌数的对数(LgC)与LgOD555 nm线性关系的标准曲线为LgC=(1.0439±0.0200)LgOD555 nm+(8.0565±0.0125),相关系数R²=0.9965,线性检测范围为7.8×106~2.5×108 CFU/ml;ATP生物发光法以ATP产生的相对发光度值(RLU)为计数依据,LgC与LgRUL线性关系的标准曲线为LgC=(0.9590±0.0065)LgRLU+(0.9949±0.0366),相关系数R²=0.9994,线性检测范围为1.0×104~3.0×108 CFU/ml;高通量生长曲线法以生长曲线达到拐点的时间(Ts)为计数依据,LgC与Ts线性关系的标准曲线为LgC=?(0.8727±0.0230)Ts+(9.0128±0.1572),相关系数R²=0.9924,线性检测范围为1.0×100~1.0×107 CFU/ml。用3种方法对实际菌液测量并与平板计数法比较表明,ATP生物发光法与高通量生长曲线法有很好的准确性,MTT比色法准确度稍差,而高通量生长曲线法有最宽的线性范围,也最适合高通量测定。  相似文献   

MTT法检测大鼠外周血淋巴细胞增殖反应探讨与应用   总被引：1，自引：1，他引：0  

梁宏伟  朱雯宇  冯波  王建舫  张涛  穆祥 《中国农学通报》2016,32(26):21-26

为了更精准简捷地评价机体的细胞免疫状态,采用MTT比色分析法检测大鼠外周血淋巴细胞的增殖能力,并对试验的最适条件进行了探讨,然后将该法应用到Transwell共培养体系中以评价H9N2 AIV感染内皮细胞后对跨内皮淋巴细胞增殖能力的影响。结果表明：在96孔细胞培养板中接种密度为1.25×10⁶个/mL的淋巴细胞于含10% FBS的RPMI 1640培养基中培养的最适时间为48 h。感染病毒后,跨内皮淋巴细胞的增殖明显受抑制;内皮完整状态下,淋巴细胞和病毒共孵育后,活病毒的量降低,而当内皮不存在或被病毒感染后,这一现象消失。结果证明作为机体细胞免疫功能评价指标之一的淋巴细胞增殖能力可采用改良后的MTT法检测。  相似文献   

哲罗鱼胰岛素样生长因子-II的原核表达与活性分析     

吴秀梅  徐黎明  赵景壮  刘淼  曹永生  卢彤岩  尹家胜 《中国水产科学》2015,22(2):243-249

在鱼类胚胎发育过程中,胰岛素样生长因子-II(insulin-like growth factors-II,IGF-II)起着关键的作用。本研究根据Gen Bank收录的鲑鳟鱼IGF-II基因序列设计引物,以哲罗鱼(Hucho taimen)肝RNA提取物为模板,利用RT-PCR方法扩增出哲罗鱼IGF-II基因开放阅读框。将目的基因IGF-II与原核表达载体p SUMO连接构建出重组表达载体p SUMO-IGF。将该重组质粒转化到大肠杆菌Rosetta中进行目的蛋白的诱导表达。SDS-PAGE电泳分析显示,约在40 k D处含有清晰的条带,与预期结果相符;目的蛋白以包涵体形式存在。对包涵体进行变性/复性后获得较纯的目的蛋白,利用ELISA和MTT方法对目的蛋白进行免疫学活性及生物学活性分析。ELISA结果显示该目的蛋白能够与商品化的抗鲑鳟鱼IGF-II的抗体发生特异性反应,并且呈现抗原浓度依赖性,该结果说明本研究获得了具有良好免疫原性的IGF-II蛋白;MTT方法测定IGF-II蛋白对鲤(Cyprinus carpio)上皮细胞(epitheliaoma papulosum cyprini,EPC)和虹鳟(Oncorhynchus mykiss)性腺细胞(rainbow trout gonad,RTG-2)的增殖效果来鉴定IGF-II蛋白的生物学活性,结果显示所表达的哲罗鱼IGF-II蛋白能够有效的刺激EPC细胞和RTG-2细胞增殖。该结果表明利用原核表达系统获得的哲罗鱼IGF-II蛋白具有良好的生物学活性。本研究为哲罗鱼的生长模式和生长繁殖的研究奠定了基础。  相似文献   

玉屏风散对鸡脾脏淋巴细胞体外增殖的影响   总被引：1，自引：0，他引：1  

林丽华  郑雅  钟文平  何玉琴 《福建畜牧兽医》2009,31(4):1-2

将不同浓度的玉屏风散加入到鸡脾脏淋巴细胞中,培养24、48、72 h,采用MTT法观察其对鸡脾脏淋巴细胞增殖的影响.结果表明,不同浓度的玉屏风散均能协同ConA促进鸡脾脏淋巴细胞增殖的作用,并以500 mg/mL的玉屏风散培养48 h增殖作用最强.  相似文献   

Wildfire risk in the wildland–urban interface: A simulation study in northwestern Wisconsin     

Avi Bar Massada  Volker C. Radeloff  Susan I. Stewart  Todd J. Hawbaker 《Forest Ecology and Management》2009

The rapid growth of housing in and near the wildland–urban interface (WUI) increases wildfire risk to lives and structures. To reduce fire risk, it is necessary to identify WUI housing areas that are more susceptible to wildfire. This is challenging, because wildfire patterns depend on fire behavior and spread, which in turn depend on ignition locations, weather conditions, the spatial arrangement of fuels, and topography. The goal of our study was to assess wildfire risk to a 60,000 ha WUI area in northwestern Wisconsin while accounting for all of these factors. We conducted 6000 simulations with two dynamic fire models: Fire Area Simulator (FARSITE) and Minimum Travel Time (MTT) in order to map the spatial pattern of burn probabilities. Simulations were run under normal and extreme weather conditions to assess the effect of weather on fire spread, burn probability, and risk to structures. The resulting burn probability maps were intersected with maps of structure locations and land cover types. The simulations revealed clear hotspots of wildfire activity and a large range of wildfire risk to structures in the study area. As expected, the extreme weather conditions yielded higher burn probabilities over the entire landscape, as well as to different land cover classes and individual structures. Moreover, the spatial pattern of risk was significantly different between extreme and normal weather conditions. The results highlight the fact that extreme weather conditions not only produce higher fire risk than normal weather conditions, but also change the fine-scale locations of high risk areas in the landscape, which is of great importance for fire management in WUI areas. In addition, the choice of weather data may limit the potential for comparisons of risk maps for different areas and for extrapolating risk maps to future scenarios where weather conditions are unknown. Our approach to modeling wildfire risk to structures can aid fire risk reduction management activities by identifying areas with elevated wildfire risk and those most vulnerable under extreme weather conditions.  相似文献   

Study on the Inhibition of Fermented Soybean to Cancer Cells     

LU Yan  WANG Wei  SHAN Yi  E Zhiqiang  WANG Liqun 《东北农业大学学报(英文版)》2009,16(1):25-28

In the experiment, the inhibition of isoflavones extracted from soybean and tempe to SP2/0 and Hela cells was studied, and the inhibition rate of each unit for cancer cells was also studied. The results showed that the inhibition rate of tempe isoflavones to SP2/0 was 96.9% and to Hela cells was 69.5% when the concentration was 20μg·mL^-1. In the same condition, the inhibition rate of soybean isoflavones was 83.16% and 60.5%. With the decline of concentration, the inhibition rate decreased. The inhibition of isoflavones to SP2/0 did not exist when the concentration was 5-1.25μg·mL^-1.  相似文献