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为探讨牛皮消多糖（CAP）对体外抗新城疫病毒（NDV）能力的影响，本试验用水提醇沉法提取牛皮消多糖，通过苯酚硫酸法和红外光谱对牛皮消多糖进行检测，采用MTT法测定牛皮消多糖对鸡胚成纤维细胞（CEF）的安全浓度，多糖的安全浓度统一为625 μg/mL，选择安全浓度范围内的（39.06~625 μg/mL）5种浓度的各级多糖，用3种加药方式（先加多糖后接种病毒、先接种病毒后加多糖、多糖和病毒感作后加药）检测牛皮消多糖对NDV的阻断、抑制及直接杀灭作用。结果显示，先接病毒后加多糖组和多糖与病毒感作后加药组CAP50、CAP30、CAPt、CAP80和CAP70均能显著抑制NDV感染CEF的能力，两组各级多糖抑制率分别为131.23%、110.12%、108.15%、100.24%、93.07%和61.76%、43.73%、44.51%、29.02%、37.45%；先加多糖后接病毒组CAP50、CAP30能显著抑制NDV感染CEF的作用（P<0.05），阻断作用下对NDV的抑制率可达到15.82%、8.33%，其余各组在安全浓度范围内对NDV无抑制作用。各组中药牛皮消多糖均有较好的抗NDV活性，其中对NDV的抑制作用和直接杀灭作用强于对其阻断作用。CAP50及CAP30的抗NDV活性较强，可作为进一步研究材料并具备一定的研究价值。 相似文献
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为研究列当多糖抗新城疫病毒(NDV)活性,用水提醇沉法提取列当总多糖及4种分级多糖,并通过苯酚硫酸法及红外光谱对列当多糖进行检测。通过MTT法测定各级多糖对鸡胚成纤维细胞(CEF)的安全浓度。以3种加药方式(先加多糖后接种病毒、先接种病毒后加多糖、多糖和病毒感作后加)加药,检测列当多糖对新城疫病毒的阻断作用、抑制作用及直接杀灭作用。结果表明,列当总多糖及4种分级多糖均具有抗NDV活性,其中对NDV的抑制作用及阻断作用强于对NDV的直接杀灭作用。50%、80%梯度醇沉的列当多糖(OSP50、OSP80)及总多糖(OSPt),阻断作用下的病毒抑制率分别为19.50%、33.10%和22.30%,抑制作用下的病毒抑制率分别为19.00%、21.90%和22.60%,杀灭作用下对NDV的抑制率分别为,14.70%、17.50%和13.30%,其余各组多糖阻断作用、抑制作用及杀灭作用下的病毒抑制率均低于上述3组多糖。说明OSP50、OSP80及OSPt的抗NDV活性较好,可以作为进一步研究的材料。 相似文献
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为探讨牛皮消多糖(CAP)对体外抗新城疫病毒(NDV)能力的影响,本试验用水提醇沉法提取牛皮消多糖,通过苯酚硫酸法和红外光谱对牛皮消多糖进行检测,采用MTT法测定牛皮消多糖对鸡胚成纤维细胞(CEF)的安全浓度,多糖的安全浓度统一为625μg/mL,选择安全浓度范围内的(39.06～625μg/mL)5种浓度的各级多糖,用3种加药方式(先加多糖后接种病毒、先接种病毒后加多糖、多糖和病毒感作后加药)检测牛皮消多糖对NDV的阻断、抑制及直接杀灭作用。结果显示,先接病毒后加多糖组和多糖与病毒感作后加药组CAP_(50)、CAP_(30)、CAP_t、CAP_(80)和CAP_(70)均能显著抑制NDV感染CEF的能力,两组各级多糖抑制率分别为131.23%、110.12%、108.15%、100.24%、93.07%和61.76%、43.73%、44.51%、29.02%、37.45%;先加多糖后接病毒组CAP_(50)、CAP_(30)能显著抑制NDV感染CEF的作用(P0.05),阻断作用下对NDV的抑制率可达到15.82%、8.33%,其余各组在安全浓度范围内对NDV无抑制作用。各组中药牛皮消多糖均有较好的抗NDV活性,其中对NDV的抑制作用和直接杀灭作用强于对其阻断作用。CAP_(50)及CAP_(30)的抗NDV活性较强,可作为进一步研究材料并具备一定的研究价值。 相似文献
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《中国兽医学报》2017,(12)
为了研究维药阿里红各活性成分抗鸡新城疫病毒(NDV)的活性,本试验分别提取了阿里红多糖、多酚、黄酮、生物碱、挥发油并测定其含量。通过MTT法测定阿里红各活性成分对鸡胚成纤维细胞的最大安全浓度。以3种加药方式将药物与NDV加入已制备好的鸡胚成纤维细胞上,即先加药后接种病毒,先接种病毒后加药,病毒和药物混合作用后加入,以检测各组分对NDV的阻断作用、抑制作用以及直接杀灭作用。结果表明,多糖、多酚、黄酮、生物碱、挥发油均有抗病毒活性,其中多糖对NDV的直接杀灭作用及抑制作用强于对NDV的阻断作用。多糖、多酚、黄酮、生物碱、挥发油在抑制作用下的病毒抑制率分别为49.12%,39.64%,42.45%,40.00%,18.59%;杀灭作用下的病毒抑制率分别为45.35%,35.71%,38.57%,35.71%,41.07%;阻断作用下的病毒抑制率分别为15.58%,8.57%,32.98%,43.11%,31.42%。综上可知,维药阿里红的活性成分多糖和生物碱的抗NDV活性较好,可以作为进一步研究的材料。 相似文献
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为研究一枝蒿的多糖、黄酮、多酚、生物碱、挥发油的抗鸡新城疫病毒的活性,多糖采用水煎醇沉法、黄酮采用浸提法、多酚采用超声法、生物碱采用超生辅助提取方法、挥发油采用水蒸气蒸馏法提取并测定含量。通过MTT法测定一枝蒿各成分对鸡成纤维细胞的安全浓度。在安全浓度以下5个浓度梯度范围内新城疫病毒(NDV)与药物以3种方式加到单层鸡成纤维细胞(CEF)上,即先加药后攻毒(阻断作用)、先攻毒后加药(抑制作用)、病毒和药物同时作用一段时间之后同时加入(直接杀灭作用),观察细胞的病变程度。细胞培养72h后用MTT法测定细胞D值的变化,用于评价药物对NDV感染细胞能力的影响。结果显示:多糖、多酚、黄酮、生物碱、挥发油均有一定程度的抗病毒活性,其中多糖对新城疫病毒的阻断作用及直接杀灭作用强于抑制作用。它们在抑制作用下的病毒抑制率分别为44.30%,11.00%,31.00%,12.94%和42.35%;直接杀灭作用下的病毒抑制率分别为27.27%,10.64%,22.85%,24.93%和10.38%;阻断作用下的病毒抑制率为33.33%,18.90%,35.19%,25.56%和40.74%。综上可知,多糖的抗鸡新城疫活性较好,其次是挥发油,可以作为进一步的研究材料。 相似文献
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维药阿魏根多糖体外抗新城疫病毒的作用 总被引：1，自引：0，他引：1
用水提醇沉法得到阿魏根总多糖FSPt和4种分级多糖FSP_(30)、FSP_(50)、FSP_(70),FSP_(80)用苯酚-硫酸法及红外光谱对阿魏根多糖进行检测。用MTT法测定各多糖对鸡胚成纤维细胞(CEF)的安全浓度,选择安全浓度范围内5种浓度的各多糖与新城疫病毒(NDV)用3种不同的加药方式(先加多糖后接种病毒、先接种病毒后加多糖、多糖和病毒感作后加)加入到鸡胚成纤维细胞(CEF)中,用MTT法测定各多糖对NDV感染CEF能力的影响。先加多糖后接病毒组和先接病毒后加多糖组FSP_(30)、FSP_(50)、FSP_(70)和FSPt均能显著抑制NDV感染CEF;多糖与病毒感作后加组FSP_(70)和FSP_(80)有显著抑制NDV感染CEF的作用(P0.05)。结论:新疆阿魏根多糖具有较好的抗NDV作用,其中FSP_(30)和FSP_(50)对病毒的抑制率较高,FSP_(70)在3种加药方式中均表现出显著且稳定的抗NDV作用,综合考虑,FSP_(50)具有很好的研发前景。 相似文献
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为评价新兽药"芪蓝饮"及其不同提取成分的体外抗病毒活性,本试验通过细胞病变抑制法和MTT比色法检测了各提取成分对新城疫病毒(NDV)感染鸡胚成纤维细胞(CEF)过程的影响。结果显示,在3种加药方式中"芪蓝饮"原药(QLY)直接杀灭NDV的作用最强,其抑制率最高为128.85%;原药总多糖(TPS)干扰NDV的增殖作用最强,其抑制率最高为130.23%;原药总萜类(TT)直接杀灭NDV的作用最强,其抑制率最高为146.15%;板蓝根总生物碱(RIA)及其水溶性部位(RIAw)和酸性部位(RIAa)对抗NDV的方式是多环节的。综上所述,"芪蓝饮"及其提取成分表现出较强的干扰NDV增殖和直接杀灭NDV的作用,而阻断作用稍弱。 相似文献
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甘草多糖和甘草酸对NDV感染鸡胚成纤维细胞能力的影响 总被引：1，自引：0，他引：1
从甘草中提取甘草多糖和甘草酸,进行抗病毒活性比较。用MTT法比较了它们对鸡胚成纤维细胞(CEF)生长及以三种加药方式(先药后毒、先毒后药、药毒同时)抵抗新城疫病毒(NDV)感染细胞的影响,并用平均病毒抑制率比较甘草多糖和甘草酸的抗病毒活性。结果表明,先药后毒,甘草多糖能显著抑制NDV感染细胞(P0.05),平均病毒抑制率为60.6%;先毒后药,甘草多糖和甘草酸都能显著抑制NDV感染细胞(P0.05),其中甘草多糖的平均病毒抑制率为74.28%,甘草酸为62.3%。通过平均病毒抑制率比较,表明甘草多糖的抗病毒活性优于甘草酸。 相似文献
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ADELIJIANG Wusiman DELIBAIER NUERAILI Maitiniyazi BUHELIQIANMU Yiming SAIFUDING Abula 《中国畜牧兽医》2016,43(7):1869-1875
To research on polysaccharides from Lagenaria siceraria (Molina) Standl.against Newcastle disease virus (NDV).Total polysaccharide and four different solvent extractions of Lagenaria siceraria (Molina) Standl.were extracted by the methods of hot water extraction and ethanol precipition in this study.The content of polysaccharide was measured by phenol-sulfuric acid method and infrared spectroscopy method.The safe concentration and growth of chick embryo fibroblast (CEF) were assayed by MTT method,in order to facilitated the comparison under the same level,the safe concentration was united as 78.125 μg/mL.Under the safety range of concentration,detected the block-virus activity,anti-virus activity and virus-killing activity of polysaccharides through the ways of pre-adding polysaccharides,post-adding polysaccharides and adding polysaccharides with NDV.The results showed that the direct inactivation and propagation inhibition activity of total polysaccharide and four different solvent extractions were stronger than anti-absorption function.Anti-virus inhibition rate of 70%,80% gradient alcohol precipitation of polysaccharides from Lagenaria siceraria (Molina) Standl.(LSP70,LSP80) and total polysaccharide (LSPt)were 40.41%,44.54% and 61.85%,virus-killing inhibition rate were 44.74%,58.76% and 59.38%.LSP80 had the highest virus inhibition rate as 37.14% in the block-virus activity of those five polysaccharides.In summary,70%,80% gradient alcohol precipitation and total polysaccharide in polysaccharides possessed better activity and would be as the materials for further research. 相似文献
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To investigate the effects of Ferula sinkiangensis polysaccharides (FSP) on cell proliferation of chicken splenic lymphocytes and peripheral lymphocyte in vitro. By the method of water extract-alcohol precipitation,total Ferula sinkiangensis polysaccharide (FSPt) and 30%,50%,70% gradient alcohol precipitation of polysaccharides (FSP30,FSP50 and FSP70) were extracted, and detected by phenol-sulfuric acid method and infrared spectroscopy method. The safe concentration of all classifications of polysaccharide were detected and the effect of FSP on lymphocyte proliferation of chicken spleen and peripheral blood were measured by MTT method. The results showed that the absorption peak of FSP were conformed to belong by polysaccharide compounds. Peripheral blood T lymphocyte proliferation test indicated that FSPt,FSP30 and FSP70 alone or cooperated with PHA could stimulate T lymphocyte proliferation under a certain concentration,and FSPt showed a better proliferation effect. Spleen B lymphocyte proliferation test showed that FSPt,FSP30 and FSP70 alone or cooperated with LPS could stimulate B lymphocyte proliferation under a certain concentration,FSPt and FSP70 showed a better proliferation effect. In conclusion,FSP showed the strong effect on lymphocyte proliferation of chicken spleen and peripheral blood in vitro,and that would be the materials for further research. 相似文献
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为研究新疆阿魏多糖（FSP）对鸡外周血T淋巴细胞和脾脏B淋巴细胞增殖作用的影响。本研究用水提醇沉法得到阿魏总多糖（FSPt）和3种分级多糖（FSP30、FSP50、FSP70）,并通过苯酚-硫酸法和红外光谱对FSP进行检测。通过MTT法测定各级FSP的安全浓度及其对鸡外周血T淋巴细胞及脾脏B淋巴细胞的增殖能力。结果显示,提取的FSP均具有多糖的特征峰,为多糖类化合物。外周血T淋巴细胞增殖试验表明,除FSP50外,所有多糖在一定浓度下均能单独和协同PHA刺激T淋巴细胞增殖,其中FSPt的增殖作用较强。脾脏B淋巴细胞增殖试验表明,除FSP50外,所有多糖在一定浓度下均能单独和协同LPS刺激B淋巴细胞增殖,其中FSP70和FSPt增殖作用较强。综上所述,各级FSP中,FSPt对脾脏B淋巴细胞和外周血T淋巴细胞的增殖作用最强,可以作为进一步研究的材料。 相似文献
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本研究旨在对鸡新城疫病毒(Newcastle disease virus,NDV)LaSota株在BHK-21细胞上的增殖条件进行优化,为实现应用细胞系生产NDV提供参考。在确定BHK-21细胞TPCK-胰酶耐受浓度的基础上,在培养基中添加不同浓度的TPCK-胰酶、新生牛血清及调节不同pH,通过对细胞培养液细胞半数感染量(TCID50)的测定,确定最适胰酶浓度、血清浓度及pH;通过固定接毒量而添加不同数量细胞或固定细胞数量而接种不同量的病毒,确定最适细胞量及接毒量;将BHK-21细胞接种NDV LaSota株后,通过测定不同时间细胞上清液TCID50绘制增殖曲线,确定最佳收毒时间;应用转瓶对该病毒进行扩大培养,并进行TCID50、鸡胚半数感染量(EID50)与血清凝集价(HA)的测定。结果显示,NDV LaSota株在BHK-21细胞上增殖的最适TPCK-胰酶浓度为3μg/mL,最适接毒量为106.375 EID50,细胞数量为1×10~6个,培养基pH为7.2,且可用无血清培养基进行培养,BHK-21细胞接种NDV LaSota株后最佳收毒时间为34～36 h。应用BHK-21细胞采用转瓶培养的方式对NDV LaSota株进行增殖,结果显示,细胞培养液TCID50值为107.0/0.1 mL,EID50为107.5/0.1 mL,HA效价为1∶256。因此,应用以上条件增殖培养NDV LaSota株,完全适用于该病毒的规模化生产,且符合现行的兽医生物制品与检验制造规程。 相似文献
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LI Xiao-wen LIU You-ming LIANG Guo-zhi MEI Min-min HUANG Wen-jing LI Wen-feng HUANG Shu-jian LI Zhi-li 《中国畜牧兽医》2017,44(7):1925-1933
In order to study the relationship and evolution between the latest Newcastle disease virus (NDV) epidemic strain and the vaccine strain (Mukteswar,La Sota and Clone 30 strains),four NDV epidemic strains were initially isolated and identified from immunized chicken groups in Guangdong,Guangxi and Hainan provinces. Fusion (F) gene and hemaglutinin neuraminidase (HN) gene of four isolated stains were amplified,cloned and sequenced by RT-PCR method,and the homology analysis of amino acid sequences deduced by F and HN genes of NDV and other strains published in GenBank were studied, the phylogenetic tree were build,and the pathogenicity index of each strain (EID50,MDT, ICPI and IVPI) were determined. The results showed that the EID50,MDT,ICPI and IVPI of the HN-08 strain were 10-8.37/0.2 mL,58.5 h,1.78 and 2.45,that of the XX-08 strain were 10-6.50/0.2 mL,75.0 h,1.61 and 2.41,that of the YS-09 strain were 10-7.75/0.2 mL,64.5 h,1.71 and 2.38,and of the LF-09 strain were 10-7.60/0.2 mL,63.8 h,1.84 and 2.38.In this experiment,the similarity of amino acid sequences of F and HN genes was over 95.8% among four strains of NDV isolated from chicken,the similarity of four NDV strains with chicken epidemic strains (GX11/03 and GM strains),vaccine strains (Mukteswar,La Sota and Clone 30 strains) and standard virulent strain (F48E9 strain) were from 96.8% to 98.6%,86.7% to 90.6% and 88.4% to 91.3%,respectively. The results showed that four NDV epidemic strains were virulent strains,which were closely related to the GX11/03 and GM strains that were popular in China in recent years,but relatively far from the traditional vaccine strains. 相似文献
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LU Guang-fu CHEN Xiao-lan HUANG Ya-qi JIANG Chun-mao YU Jie QU Jing-wen GENG Qian-li 《中国畜牧兽医》2016,43(7):1922-1927
The experiment was conducted to study the best extract process of mulberry leaf polysaccharide,three factors and three levels orthogonal test was designed to optimize the extraction process.Three factors were considered,including ratio of solid to liquid,decoction time,the time of extracting and the yield of mulberry leaf polysaccharide was determined with phenol-sulfurie acid colorimetry as the evaluate index.Meanwhile,the influence of ethanol concentrations on mulberry leaf polysaccharide extract was studied.The results showed that decoction times could greatest affect the extraction percent of the polysaccharides.The extraction percent of polysaccharide was the highest (4.94%) when the rate of solid to liquid was 25,the time of extracting was 2 h and decoction times was 2.While the most appropriate process was 25 times for the rate of sdid to liquid,2 h for extracting and decoction times was 3,considering with practical situations.The concentration of ethanol would significantly affect polysaccharides yield and purity.When the concentration of ethanol was 70%,the yield of mulberry leaf polysaccharid was highest.When the concentration of ethanol was 40%,the content of mulberry leaf polysaccharide was highest (41.5%),significantly higher than other ethanol concentration (P < 0.05).It suggested that the higher the ethanol concentration,the lower the polysaccharide content,but the higher the yield.The optimum extraction of mulberry leaf polysaccharides were that the rate of solid to liquid was 25,extractign for 2 h,decoction 3 times and 70% ethanol precipitation. 相似文献
17.
旨在了解猪脾转移因子（TF）对新城疫病毒（NDV）弱毒疫苗LaSota株的免疫增强效果及机制。本研究分别采用不同剂量（10-2、10-3、10-4、10-5羽份） LaSota疫苗株单独（单独免疫组）、LaSota疫苗株与TF联合（联合免疫组）免疫SPF鸡,14 d后以NDV F48E9强毒攻毒,同时设立对照组（非免疫攻毒）和空白组（非免疫非攻毒）,并采用ELISA方法与蛋白质芯片技术分别测定外周血IL-4、IFN-γ与IL-12 P40浓度。结果显示,鸡免疫10-2、10-3、10-4、10-5羽份时疫苗攻毒保护率和半数保护量（PD50）如下：单独免疫组分别为100%、55%、0%、0%和0.000 8羽份,联合免疫组分别为100%、75%、0%、0%和0.000 5羽份,对照组和空白组死亡率分别为100%和0%。免疫10-3羽份疫苗后,联合免疫组IL-4和IFN-γ含量均高于其他组,并于第7、14天差异极显著（P<0.01）;攻毒后,联合免疫组IL-4、IFN-γ和IL-12 P40含量均高于其他组,IL-4于第1、14天,IFN-γ于第1、3天,IL-12 P40于第1天差异极显著（P<0.01）。综上表明,TF可增强IFN-γ、IL-12介导的细胞免疫和IL-4介导的体液免疫,提高LaSota株弱毒疫苗的免疫保护率,降低疫苗PD50;在抵抗NDV F48E9强毒攻击时,联合免疫组的免疫保护率明显高于单独免疫组。 相似文献
18.
The study was aimed to observe the effects of Codonopsis pilosula polysaccharides(CPPS) and selenium-Codonopsis pilosula polysaccharides5(sCPPS5) on the immune functions of immunosuppressive mice induced by cydophosphamide (CY).96 mice were randomly divided into 8 groups:Normal control group,CY model group,CY+CPPS (low,middle and high dose groups) and CY+sCPPS5 (low,middle and high dose groups).Mice in normal control group and CY model group were lavaged with normal saline,the mice in other groups were lavaged with CPPS and sCPPS5 by 0.2,0.4 and 0.6 mg/g,respectively.Administration was carried out once a day for 10 successive days.On the 11th day,mice were manufactured immunosuppressed by a three-day intraperitoneal injection of 0.04 mg/g CY,except for the normal control group.After administrated with drugs for 24 h,the immunological indexes of the mice were detected.The results showed that spleen and thymus indexes of CPPS and sCPPS5 groups were significantly higher than that of CY group (P < 0.05).The A570 nm when stimulated with ConA and LPS,serum IgG and IgM contents of CPPS and sCPPS5 groups were significantly higher than that of CY group (P < 0.05) and sCPPS5 possessed better efficacy than CPPS.In conclusion,CPPS and sCPPS5 could strengthen immune activity of the immunosuppressive mice,but sCPPS5 was better than CPPS and selenylation modification could improve the immune activity of polysaccharide. 相似文献