Aim To study whether antivenom from laying hens can be used for the detection of venom antigens, Methods Chickens （white Leghorn） were immunized with detoxicated king cobra venom by formaldehyde and egg yolk immunog...Aim To study whether antivenom from laying hens can be used for the detection of venom antigens, Methods Chickens （white Leghorn） were immunized with detoxicated king cobra venom by formaldehyde and egg yolk immunoglobulin （IgY） isolated from yolk; IgY was labelled with the horseradish peroxidase （HRP）. Experimental condition and parameters were determined by chessboard test. The specificity, sensitivity, precision, and stability of this method were assayed in the experiment. Results This method could detect as low as 32 μg· L^-1 of the king cobra antigens. A good linear relation was found within 32 ～ 750 μg· L^-1 of king cobra venom concentrations （ r = 0. 963）. There was no cross reactivity for the reagents with Agkistrodon acutus Guenther venom or Vipera russelli siamensis Smith venom;slight cross reactivity .with Bungarus multicinctus Blyth venom or Bungarus fasciatus Chmeider venom; and notable cross reactivity with cobra venom. The average intra-assay relative standard deviation （RSD） was 1% - 3%, and the inter-assay RSD was less than 8%. The reagents （including IgY and HRP-IgY） were stable; no differences （P 〉 0.05） were observed for the detection of venom antigens when the reagents were stored at 37 ℃ up to 6 d. Conclusion IgY is a good reagent for diagnosis of snakebite after eliminating the genus cross reactivity.展开更多
目的观察中华眼镜蛇毒活性组分(Ch ina cobra venomactive factor,CCVAF)对内皮细胞增殖的选择性抑制作用。方法实验分对照组和不同浓度的CCVAF组,应用MTT法分析CCVAF对牛肺主动脉内皮细胞(bovine arteria pu lmona-lis vascu lar endot...目的观察中华眼镜蛇毒活性组分(Ch ina cobra venomactive factor,CCVAF)对内皮细胞增殖的选择性抑制作用。方法实验分对照组和不同浓度的CCVAF组,应用MTT法分析CCVAF对牛肺主动脉内皮细胞(bovine arteria pu lmona-lis vascu lar endothelial cells,BAVEC)、SD大鼠主动脉血管平滑肌细胞(smooth musc le cells,SMC)、人胚肺成纤维细胞HLF以及人支气管上皮细胞HBE生长增殖的影响;并运用直线回归分析统计对比CCVAF在不同时间点对BAVEC的IC50及上述各细胞的24 h IC50。结果CCVAF呈剂量及时间依赖性抑制内皮细胞的增殖:6、12、24、48、72 h其IC50分别为4.955、4.932、2.443、3.048、3.133 mg.L-1,24 h抑制作用最强;观察此时浓度与抑制作用的关系,0.625、1.25、2.5、5、10、20 mg.L-1,抑制率分别是4.41%、30.94%、61.81%、86.17%、96.05%、98.68%,10 mg.L-1抑制率即达高峰,且各浓度组抑制作用与对照组相比差异有显著性(P<0.05)。CCVAF抑制内皮细胞增殖具有一定的选择性,作用24 h后,抑制BAVEC生长的作用明显高于其它3种细胞,对BAVEC、HLF、HBE、SMC的24 h IC50分别为2.443、22.233、35.318、32.810 mg.L-1。结论CCVAF能够选择性抑制内皮细胞增殖,这在抗血管生成及其靶向治疗研究中可能具有重要意义。展开更多
文摘Aim To study whether antivenom from laying hens can be used for the detection of venom antigens, Methods Chickens （white Leghorn） were immunized with detoxicated king cobra venom by formaldehyde and egg yolk immunoglobulin （IgY） isolated from yolk; IgY was labelled with the horseradish peroxidase （HRP）. Experimental condition and parameters were determined by chessboard test. The specificity, sensitivity, precision, and stability of this method were assayed in the experiment. Results This method could detect as low as 32 μg· L^-1 of the king cobra antigens. A good linear relation was found within 32 ～ 750 μg· L^-1 of king cobra venom concentrations （ r = 0. 963）. There was no cross reactivity for the reagents with Agkistrodon acutus Guenther venom or Vipera russelli siamensis Smith venom;slight cross reactivity .with Bungarus multicinctus Blyth venom or Bungarus fasciatus Chmeider venom; and notable cross reactivity with cobra venom. The average intra-assay relative standard deviation （RSD） was 1% - 3%, and the inter-assay RSD was less than 8%. The reagents （including IgY and HRP-IgY） were stable; no differences （P 〉 0.05） were observed for the detection of venom antigens when the reagents were stored at 37 ℃ up to 6 d. Conclusion IgY is a good reagent for diagnosis of snakebite after eliminating the genus cross reactivity.
文摘目的观察中华眼镜蛇毒活性组分(Ch ina cobra venomactive factor,CCVAF)对内皮细胞增殖的选择性抑制作用。方法实验分对照组和不同浓度的CCVAF组,应用MTT法分析CCVAF对牛肺主动脉内皮细胞(bovine arteria pu lmona-lis vascu lar endothelial cells,BAVEC)、SD大鼠主动脉血管平滑肌细胞(smooth musc le cells,SMC)、人胚肺成纤维细胞HLF以及人支气管上皮细胞HBE生长增殖的影响;并运用直线回归分析统计对比CCVAF在不同时间点对BAVEC的IC50及上述各细胞的24 h IC50。结果CCVAF呈剂量及时间依赖性抑制内皮细胞的增殖:6、12、24、48、72 h其IC50分别为4.955、4.932、2.443、3.048、3.133 mg.L-1,24 h抑制作用最强;观察此时浓度与抑制作用的关系,0.625、1.25、2.5、5、10、20 mg.L-1,抑制率分别是4.41%、30.94%、61.81%、86.17%、96.05%、98.68%,10 mg.L-1抑制率即达高峰,且各浓度组抑制作用与对照组相比差异有显著性(P<0.05)。CCVAF抑制内皮细胞增殖具有一定的选择性,作用24 h后,抑制BAVEC生长的作用明显高于其它3种细胞,对BAVEC、HLF、HBE、SMC的24 h IC50分别为2.443、22.233、35.318、32.810 mg.L-1。结论CCVAF能够选择性抑制内皮细胞增殖,这在抗血管生成及其靶向治疗研究中可能具有重要意义。
