摘要
畜禽疾病特别是病毒性疾病仍然是阻碍养殖业健康发展的瓶颈,疫苗是防治疾病最有效的方法。但是,近几年由于免疫抑制性病毒的普遍存在,预防工作变得异常艰难,国内外学者一直致力于研究提高疫苗的特异性和非特异性免疫增强作用。中药是我国传统文化的结晶,是中华民族的文化瑰宝,在畜禽疾病的防治过程中起过重要的作用。中药免疫增强剂对机体的细胞免疫和体液免疫功能具有增强或调节作用,可提高机体的抗病力和抵抗力,显示了显著增强机体免疫的作用。本研究以临床上常常发生的网状内皮组织增生症病毒(REV)感染雏鸡,造成一定程度的免疫抑制,建立试验动物模型,比较中药提取物对不同免疫状态下鸡的免疫调节作用,并对主要成分多糖进行提取分离鉴定和分子量测定,为下一步多糖的结构和功能研究奠定基础。
1中药提取物免疫调节作用的研究
1.1中药提取物和盐酸左旋咪唑对鸡的免疫调节作用
分别在临床健康鸡和在1日龄感染网状内皮增生症病毒(REV)诱发了免疫抑制的鸡测试了中药提取物和盐酸左旋咪唑对鸡的免疫调节作用,以及对鸡体重、免疫器官的影响。结果表明,在饮水中添加50ppm的盐酸左旋咪唑,无论在健康鸡还是免疫抑制鸡对NDV、H5-AIV灭活疫苗免疫后的HI抗体水平、体重和免疫器官指数都没有显著影响(p>0.05)。由黄芪、党参、淫羊藿和甘草组成的提取物饲喂健康鸡对NDV和H5-AIV免疫后的HI抗体水平也没有显著影响(p>0.05)。但是在REV诱发了免疫抑制的鸡,相对于不喂中药提取物的对照组,该中药提取物可显著提高对NDV、H5-AIV的HI抗体水平和免疫器官指数(p<0.01),虽然仍显著低于未经REV感染的对照鸡。该研究表明,由黄芪、党参、淫羊藿和甘草组成的提取物对REV诱发的免疫抑制状态下的鸡有显著的免疫增强作用,盐酸左旋咪唑没有免疫增强作用。
1.2不同组合中药提取物对鸡的免疫调节作用
分别在临床健康鸡和在1日龄感染网状内皮增生症病毒(REV)诱发了免疫抑制的鸡测试了不同浓度黄芪提取物和由黄芪、党参、淫羊藿、甘草组成的不同组合的中药提取物对鸡的体液免疫调节作用,以及对鸡体重、免疫器官的影响。结果表明,不同浓度的单一黄芪提取物对NDV和H5-AIV免疫后的HI抗体滴度没有显著影响,对鸡的体重、免疫器官指数也没有显著影响(P>0.05)。由黄芪、党参、淫羊藿和甘草混合物组成的中药提取物,在健康鸡对NDV和H5-AIV灭活疫苗免疫后的HI抗体滴度没有显著影响。但是在REV诱发了免疫抑制的鸡,该混合中药提取物可显著提高对NDV和H5-AIV的HI抗体滴度(p<0.01),虽然仍显著低于未经REV感染的对照鸡。该中药制剂还可以显著增加胸腺和法氏囊指数,即缓和由REV诱发的萎缩。该研究表明,单一黄芪不能提高由REV诱发的免疫抑制鸡的抗体水平,黄芪、党参、淫羊藿和甘草组成的中药提取物对REV诱发的免疫抑制鸡具有显著的免疫增强作用。
13不同提取方法中药提取物对鸡的免疫调节作用
分别在临床健康鸡和1日龄感染网状内皮组织增生症病毒(REV)诱发了免疫抑制的鸡测试了经不同提取方法获得的中药提取物的免疫调节作用,并研究水溶性提取物对不同来源鸡的免疫调节作用。结果表明,在饮水中添加由黄芪、党参、淫羊藿和甘草组成的提取物,无论是水溶性提取物(水提取物和乙醚提取后的水提取物)还是脂溶性提取物(乙醚提取物),对健康鸡NDV和H5-AIV灭活疫苗免疫后HI抗体水平均无显著影响(P>0.05),水溶性提取物对REV诱发的免疫抑制鸡的NDV和H5-AIV灭活疫苗免疫后HI抗体水平有显著影响(p<0.01),可显著提高其抗体水平。但是,由同样药物组成的脂溶性提取物却没有免疫增强作用(P>0.05)。中药提取物对健康鸡和SPF来源继续在SPF下饲养的鸡对NDV和H5-AIV灭活疫苗免疫后的HI抗体滴度没有显著影响(p>0.05)。但是在REV诱发了免疫抑制的鸡和SPF来源在非SPF条件下饲养的鸡,该中药提取物可显著提高对NDV和H5-AIV的HI抗体滴度(p<0.01),虽然仍显著低于未经REV感染的对照鸡。
1.4不同分子量段中药提取物对鸡的免疫调节作用
用网状内皮增生症病毒(REV)感染海蓝褐公鸡和SPF来源的鸡诱发免疫抑制,在饮水中添加中药提取物或不同分子量段的中药提取物,测试其对鸡的免疫调节作用。结果表明,同样由黄芪、党参、淫羊藿和甘草组成的提取物和分子量段在1-10万道尔顿的提取物,不论是先混合、提取再超滤、浓缩还是先分别提取、超滤再浓缩、混合可显著提高感染REV鸡的NDV和H5-AIV的HI抗体滴度(p<0.01),虽然仍显著低于未经感染REV的对照鸡。但分子量小于1万道尔顿和大于10万道尔顿的提取物,对感染REV的免疫抑制鸡没有显著免疫增强作用(p>0.05)。试验结果还表明,中药提取物和分子量在1-10万道尔顿的提取物对SPF来源的鸡在非SPF环境下饲养有显著的免疫增强作用(p<0.05),对同样是SPF来源的鸡继续在SPF环境下饲养却没有显著的免疫增强作用(p>0.05)。该研究表明,由黄芪、党参、淫羊藿和甘草组成的提取物其免疫增强作用的分子量在1-10万道尔顿之间,而对SPF来源继续在SPF环境下饲养的鸡没有显著免疫增强作用。
1.5中药提取物对鸡外周淋巴细胞转化率的影响
为了检测中药提取物对外周淋巴细胞转化率的影响,采用中药提取物对感染网状内皮组织增生症病毒(REV)鸡外周血液、胸腺和脾脏淋巴细胞转化率进行试验,建立MTT法来观察淋巴细胞的转化率。结果显示,REV显著抑制雏鸡血液、胸腺和脾脏淋巴细胞转化率(p<0.05)；中药提取物对健康雏鸡血液、胸腺和脾脏淋巴细胞转化率没有显著的影响(p>0.05),可显著提高由REV诱发的免疫抑制鸡35日龄前血液、胸腺淋巴细胞转化率和21日龄前脾脏淋巴细胞的转化率(p<0.05),对49日龄后血液、胸腺和35日龄后脾脏淋巴细胞转化率影响不明显(p>0.05)。
2多糖的提取分离鉴定和分子量测定
2.1中药提取物化学成分初步鉴定
本试验用中药提取物化学成分的鉴定方法鉴定了由黄芪、党参、淫羊藿和甘草组成的用不同方法提取的混合物的成分。结果表明,用水浸提后得到的提取物含有鞣质、多糖及其苷类、皂苷、氨基酸、多肽和蛋白质,用乙醚浸提后得到的提取物含有生物碱、黄酮及其苷类、挥发油、油脂类,用乙醚浸提后再用水浸提得到的提取物和用水浸提后得到的提取物成分相同,水浸提后的提取物用超滤膜过滤后,分子量小于1万道尔顿的提取物含有鞣酸、氨基酸、多肽和蛋白质,分子量在1-10万道尔顿的提取物中含有多糖及其苷类、皂苷、多肽和蛋白质,分子量大于10万道尔顿的提取物含有糖、多糖及其苷类、皂苷、多肽和蛋白质。水溶性成分和脂溶性成分的区别在于水溶性成分含有糖、多糖、皂苷,而脂溶性成分没有,证明多糖及其苷类和皂苷具有免疫增强作用。
2.2中药粗多糖的提取参数研究
本试验采用四因素三水平正交设计研究了提取温度(A),料液比(B),提取时间(C)及提取次数(D)对中药粗多糖得率的影响。最佳提取条件参数为A3B1C3D3,即浸提温度80℃,料液比1：10,浸提时间3h,提取次数3次。采用苯酚-硫酸法测得复方中药多糖提取物GCPS中多糖含量为48.45%,测试结果稳定可靠。苯酚-硫酸法是目前测定多糖含量较可靠的传统方法之一,本法简单快速,灵敏度高,重现性好。
2.3中药多糖的分离纯化和分子量测定
由黄芪、党参、淫羊藿和甘草组成的复方中药提取物,按Sevag法脱除蛋白质后得复方粗多糖GCPS。粗多糖分别用终浓度50%、60%和70%的乙醇沉淀,离心,干燥,得到三种多糖组分(GCPS-1、GCPS-2和GCPS-3);将分级醇沉所得多糖组分分别进行DEAE-纤维素柱层析,得三种主要多糖GCPS-1A, GCPS-2A, GCPS-3A,分别进行TOYOPEARL HW-55F凝胶柱层析,得三种主要成分GCPS-1AⅠ, GCPS-2AⅡ, GCPS-3AⅢ。经过Sephadex G-200凝胶柱层析、紫外可见分光光度分析和纸层析鉴定,表明是均一组分,不含有核酸和蛋白质杂质；并测得均一组份GCPS-1AⅠ中的多糖含量为77.6%,GCPS-2AⅡ中的多糖含量为95.2%,GCPS-3AⅢ中的多糖含量为87.3%；根据已知分子量的标准葡聚糖Dextran系列的洗脱体积得到标准曲线方程：Y=-0.1734X+0.8676,R=0.9993,并根据标准方程测得GCPS-1AⅠ, GCPS-2AⅡ及GCPS-3AⅢ的相对分子质量(Mr)为301254、94541和12453。
3结语
本研究首次应用REV作为免疫抑制因素建立试验动物模型,研究了由黄芪、党参、淫羊藿和甘草组成的提取物对不同免疫状态下雏鸡和SPF鸡的免疫调节作用,不同分子量段提取物的免疫调节作用,不同提取方法提取物的免疫调节作用,不同组合中药提取物的免疫调节作用,并与盐酸左旋咪唑进行了比较。还进一步研究了中药提取物对试验动物细胞免疫的调节作用。建立了应用REV作为测试中药提取物免疫增强作用的试验模型,证实了复方中药提取物的免疫增强作用。而且这种有效成分是水溶性的,分子量在1-10万道尔顿之间。但具体的化学成分还有待于进一步研究。
本研究还对起主要免疫增强作用的化学成分多糖进行提取、分离和鉴定,探索了多糖的最佳提取条件,测试了均一多糖的含量和分子量。
Poultry diseases, especially viral diseases, remain an obstacle to the development of animal husbandry. Vaccine is the most effective way to prevent disease. However, because of immune suppressive diseases prevalence in recent years, prevention has become exceptionally difficult. Scholars have been working to improve the immune-enhancing role of vaccine. Herbal medicine is the crystallization of Chinese traditional culture and the cultural treasures of the Chinese nation. Herbal medicine have played an important role in the process of livestock and poultry diseases prevention. The immunoenhancer of herbal medicine enhance or adjust the body cellular and humoral immunity function and enhance the body's resistance to disease, indicating a significant role in enhancing immune. reticuloendotheliosis virus (REV) infected in chickens usually in clinical practice, and resulted in a certain degree of immune suppression. In present study, animal model was established, and to compare the immunoregulatory of herbal extractions in the different immune status of chickens. The polysaccharide as a major components were isolated, identificated and determinate the molecular weight, the structure and function will be found in further study.
1 Immune regulation of herbal extraction
1.1 Effectiveness of herbal extraction and levamisole hydrochloride on immune regulation in chickens
The immune regulation of herbal extraction and levamisole hydrochloride was investigated in health chickens and immune suppressive chickens infected with REV in 1-day-old. The results showed that levamisole hydrochloride added 50mg/kg in water did not effect hemagglutination inhibition (HI) antibody titers both in the health chicken and the immune suppressive chickens after immunization with Newcastle disease virus (NDV) and H5 subtype of avian influenza virus (H5-AIV) vaccine(p>0.05), the same results was showed in body weight and the relative weight of immune organs. Herbal extraction composed of Radix Astragali, Radix Codonopis, Herba Epimedii and Radix Glycyrrizae did not effect HI antibody titers in the health chicken after immunization with NDV and H5-AIV vaccine (p>0.05), but it can significantly improve the HI antibody titers, body weight and the relative weight of immune organs in the immunosuppression chickens infected with REV(p<0.01), although the datas were still lower than the health chickens. The study showed that herbal extraction composed of Radix Astragali, Radix Codonopis, Herba Epimedii and Radix Glycyrrizae had immune enhancement effects in chickens immnunosuppressed by REV, and levamisole hydrochloride no effects.
1.2 Effectiveness of different combinations herbal extraction on immune regulation of chickens
The immune regulation of herbal extraction of different concentraction Radix Astragali and different combination composed with Radix Astragali, Radix Codonopis, Herba Epimedii and Radix Glycyrrizae was investigated in health chickens and immune suppressive chickens infected with REV in 1-day-old. The results showed that herbal extraction of different concentraction Radix Astragali was not effect on the NDV and H5-AIV of HI antibody titers in different immune status chickens, and not effect on the body weight and the relative weight of immune organs as well(p>0.05). The immune regulation of herbal extraction composed of Radix Astragali, Radix Codonopis, Herba Epimedii and Radix Glycyrrizae did not effect HI antibody titers in the health chicken immuned with NDV and H5-AIV vaccine(p>0.05), but it can significantly improve the HI antibody titers in the immunosuppression chickens infected with REV(p<0.01), although the datas were still lower than the health chickens. The herbal extraction could also increase the relative weight of immune organs. The study showed that a single Radix Astragali could not increase HI antibody titers in immunosuppressive chickens infected with REV. The herbal extraction composed of Radix Astragali, Radix Codonopis, Herba Epimedii and Radix Glycyrrizae had immune-enhancement in immunosuppressive chickens infected with REV.
1.3 Effectiveness of herbal extraction extracted by different methods on immune regulation in chickens
The immune regulation of herbal extraction extracted by different methods was investigated in health chickens and immune suppressive chickens infected with REV in 1-day-old and that of water-soluble herbal extraction was researched in different origin chickens. The results showed that the herbal extraction composed with Radix Astragali, Radix Codonopis, Herba Epimedii and Radix Glycyrrizae was added in water, no matter the herbal extraction was water-soluble extraction (water extraction and water extraction after extracted with ethyl ether) or fat-soluble extraction (ethyl ether extraction), did not effect HI antibody titers in health chicken after immuned with NDV and H5-AIV vaccine (p>0.05). The water-soluble extraction could increase the HI antibody titers of the chickens infected with REV, but the fat-soluble extraction the same composition had not immune-enhancement (P>0.05). The water-soluble extraction did not effect HI antibody titers both in the health chicken and the SPF chickens feeded in FPF environment (p>0.05), but it can significantly improve the HI antibody titers in the immunosuppressive chickens and SPF chickens feeded in non-SPF environment infected with REV(p<0.01), although the datas were still significantly lower than the health chickens.
1.4 Effectiveness of herbal extraction of different molecular weights on immune regulation in chickens
The immune regulation of herbal extraction and different molecular weght of herbal extraction was investigated in different immune status chickens and SPF chickens infected with REV in 1-day-old. The results showed that the herbal extraction and the 1-10 million Dalton molecular weight of herbal extraction composed of Radix Astragali, Radix Codonopis, Herba Epimedii and Radix Glycyrrizae, no matter the herbal extraction was mixed firstly, extracted, ultrafiltered and concentrated or extracted, ultrafiltered, concentrated respectively and then mixed, can significantly improve the H5-AIV and NDV of HI antibody titers in immunosuppressive chickens induced with REV(p<0.01), although the datas were still lower than the control without REV-infected chickens. However, less than 10,000 Dalton and greater than 100,000 Dalton molecular weight of the herbal extraction had not immune-enhancement in immunosuppressive chickens infected with REV (p>0.05). This results also showed that the herbal extraction and the 1-10 million Dalton molecular weight of herbal extraction of had immune-enhancement in immunosuppressive SPF chickens infected with REV feeded in non-SPF environment (p<0.05), but had not immune-enhancement in the same chickens feeded in SPF environment(p>0.05). The research showed that the molecular weight of the herbal extraction with immune-enhancement composed with Radix Astragali, Radix Codonopis, Herba Epimedii and Radix Glycyrrizae was between 1-10 million Dalton, and had not immunoenhancement in the SPF chickens feeded in SPF environment.
1.5 Effectiveness of herbal extraction on peripheral lymphocyte proliferation in chickens
The study was conducted to detect the effects of herbal extraction on peripheral blood, thymus and spleen lymphocytes proliferation in chicken infected with REV. MTT method was used to observe lymphatic cell proliferation. The results showed that the blood, thymus and spleen lymphocyte proliferation were immune-suppressived in chickens infected with REV (p<0.05). Herbal extraction had no effects on the lymphocytes proliferation of blood, thymus and spleen in health chicken (p>0.05), but it can increased that of the blood, thymus before 35-day-old and the spleen before 21-day-old in chickens infected with REV(p<0.05), which has not effects the lymphocyte proliferation of blood, thymus after the 49-day-old and the spleen after 35-day-old in chickens(p> 0.05).
2 Extracting, isolating, identifing and determining of molecular weight of polysaccharide
2.1 The chemical constituents identification of herbal extraction
The mixture components of herbal extraction composed with Radix Astragali, Radix Codonopis, Herba Epimedii and Radix Glycyrrizae was identified by the different motheds of chemical constituents identification. The results showed that herbal medicine was distilled by water contains tannin, sugar and their glycosides, saponins, amino acids, peptides and proteins, was extracted with ethyl ether contains alkaloids, flavonoids and their glycosides, volatile oil, fat class. The herbal medicine was distilled by water after extracted with ethyl ether firstly and was distilled by water was the same composition, after filtered with ultrafiltration membrane, the molecular weight of less than 10,000 Dalton of the herbal extraction contains tannic acid, amino acids, peptides and proteins,1-10 million Dalton contains sugars, polysaccharides, glycosides, saponins, peptides and proteins, more than 100,000 Dalton contains sugar, polysaccharides and their glycosides, saponins, peptides and proteins. The difference of fat-soluble components and water-soluble components of herbal extraction is that water-soluble components contain sugar, polysaccharides, saponins and fat-soluble components do not contain that. polysaccharides and saponins and their glycosides have a role in immune-enhancing.
2.2 The study of extracting parameters of herbal extraction polysaccharide
The effect was investigated by a four-factor three-level orthogonal design of the extraction temperature (A), ratio of herb to liquid (B), extraction time (C) and extracting times (D)of on the yield of crude polysaccharide. The best extract parameters was A3B1C3D3, that the extraction temperature was 80℃, ratio of herba to liquid was 1:10, extraction time was 3h, and extract times were 3 times. The compound polysaccharide content of GCPS was 48.45% measured by phenol-sulfuric acid method. The test results were stable and reliable. Phenol-sulfuric acid method was more reliable determination of the polysaccharide content of the traditional methods. This method was simple, rapid, high sensitivity and good reproducibility.
2.3 The study of purification and molecular weight identification of herbal extraction polysaccharides
The compound polysaccharide(GCPS) was obtained from the herbal extraction composed with Radix Astragali, Radix Codonopis, Herba Epimedii and Radix Glycyrrizae removaled of protein by Sevag method. The three polysaccharide (GCPS-1, GCPS-2 and GCPS-3) was obtained from GCPS by precipitation, centrifugation and drying using ethanol of final concentration 50%,60% and 70%. The three major polysaccharide(GCPS-1A, GCPS-2A, GCPS-3A) derived from above polysaccharide(GCPS-1, GCPS-2 and GCPS-3) was obtained with DEAE-cellulose column chromatography respectively, which were chromatographyed with TOYOPEARL HW-55F gel column and then getting the three major components of a GCPS-1A I, GCPS-2A II, GCPS-3AⅢ. After identified by Sephadex G-200 gel column chromatography, UV spectrophotometry and paper chromatography the composition (GCPS-1, GCPS-2 and GCPS-3) was homogeneous, does not contain nucleic acid and protein impurities. The polysaccharide contents of homogeneous composition (GCPS-1, GCPS-2 and GCPS-3) was 77.6%,95.2% and 87.3% respectively. The standard curve equation is obtained from series elution volume of known molecular weight dextran Dextran:Y=-0.1734X+0.8676, R=0.9993, and measured the molecular weigh(Mr) of GCPS-1AI, GCPS-2AII and GCPS-3AIII according to the standard equation:301254, 94541 and 12453.
3 Inclusion
The experimental animal model was established firstly by REV as immunosuppressive factors to study the immune enhancement of herbal extraction composed with Radix Astragali, Radix Codonopis, Herba Epimedii and Radix Glycyrrizae in different immune status chickens or SPF chicken, different molecular weight extraction, extraction with different methods, different combinations of extraction and compared with levamisole hydrochloride. Further the herbal extraction was used to research the regulation of cell-mediated immunity in experimental animals. The experimental animal model was established by REV as immunosuppressive factors to determinate the immune enhancement of herbal extraction, confirme the immune enhancement of compound herbal extraction. And the active components of the herbal extraction were water-soluble and that molecular weight is between 1-10 million Dalton. However, the specific chemical composition has yet to be studied in the future.
The polysaccharide as a major components of immune enhancement was isolated, identificated and determinated the molecular weight and content of uniformity polysaccharides for further study of the structure and function. The best extraction conditions of polysaccharide is investigated also.
引文
毕可东,崔尚金,宋晓华等.中药多糖对鸡淋巴细胞转化百分率影响的研究[J].中国预防兽医学报,2000,22(4)：289-291.
步志高,陈万芳,宋大鲁.淫羊藿-蜂胶佐剂对雏鸡细胞免疫功能的影响[J].南京农业大学学报,1996,19(1)：73-76.
陈洪亮,禚宝山,李德发.黄芪多糖对肉鸡脾淋巴细胞转化及信息分子的影响[J].中国兽医杂志,2003,39(10)：11-13.
陈洪亮,李伯涛,张家祥等.免疫活性多糖的免疫调节作用及机制研究进展.中国药理学通讯,2002,4,18(1)：249-252.
陈玉库,胡元亮,张宝康等.中药粗提物对鸡传染性法氏囊病病毒(IBDV)感染细胞能力的影响[J].江苏农业科学2002：(2)：56-58.
程相朝,陈树兴,郭曹彰等.中药免疫增强剂对MD鸡外周血液ANAE+T淋巴细胞动态的影响[J].河南农业科学,1998,1：34-36.
程相朝,龙塔.不同佐剂IBD组织灭活苗免疫效果的比较研究.动物医学进展,1999,20(3)：36-38.
储岳峰,李祥瑞,胡元亮.9种中药成分对小鼠免疫细胞活性的影响[J].南京农业大学学报,2004,27(1)：97-100.
储岳峰,颜新敏,胡元亮等.几种中药成分的免疫增强活性及其作用效果[J].中国兽医科技,2005,35(1)：67-70.
储益平,殷书梅.甲壳聚多糖降脂降胆固醇实验研究[J].时珍国医国药,1999,10(3)：178.
陈海华,许时婴,王璋.亚麻籽胶中酸性多糖和中性多糖的分离纯化.食品与发酵工程,2004,30(1)：96-100.
陈军辉,谢明勇,易秀琴等.西洋参多糖提取新工艺研究.时珍国医国药,2006,16(10)：977-979.
操继跃,窦树龙,张韶东等.地塞米松和左旋咪唑对雏鸡体液免疫应答的影响.中国兽医学报,2002,22(5)：500-502.
陈群,杨桂文,安利国.银杏白果多糖的提取、纯化和分析.中国药学杂志,2002,37(5)：331-332.
崔治中.免疫抑制性病毒多重感染在鸡群疫病发生和流行中的作用.中国家禽,2003,25(20)：1-4.
戴远威,江青艳,鲁锡祥等.三种单味补益中药对粤黄雏鸡免疫功能的影响[J].中国兽医科技,1997,27(6)：27-29.
樊福好,谢明权,林辉环.3种免疫增强剂对鸡的免疫促进效果试验.中国兽医杂志,2001,37(2)：12-3.
方一苇.具有药理活性多糖的研究现状[J].分析化学,1994,22(9)：955-960.
付秀花,王恬,顾宏伟.中草药的免疫增强作用[J].畜牧与兽医,2002,34(8)：40-43.
甘燕,石佑恩：细胞因子类基因佐剂研究进展[J].国外医学寄生虫病分册,2004,31(1)：6-11.
高学军,李庆章.黄芪多糖和香菇多糖对vMDV人工感染雏鸡红细胞免疫功能的影响[J].东北农业大学学报,2000,31(2)：169-172.
高学军,李庆章.黄芪多糖和香菇多糖对雏鸡红细胞免疫功能的影响[J].东北农业大学学报,2000,31(1)：69-71.
高云芳,陈超,张海祥等.桔梗总皂苷对大鼠高脂血症的影响.中草药,2000,31(10)：764-766.
关玉峰.人参、黄芪多糖对免疫功能影响的临床试验研究[J].航空航天医药.1997,8(2)：4-6.
管孝鞠,吴玉章.Thl极化佐剂的研究进展[J].华西药学杂志,2001,16(2)：118-120.
管孝鞠,吴玉章.人用疫苗佐剂的研究现状[J].药学进展,2001,25(4)：207-211.
郭孝武.一种提取中草药化学成分的方法-超声提取法.天然产物研究与开发.1999,11(3)：37-40.
顾承志,鲁建江,王莉等.微波提取红景天茎中总黄酮和多糖及其含量测定.安徽医药,2004,8(4)：277-278.
高丽君.白首乌可溶性多糖提取工艺、分离纯化及免疫功能研究.山东农业大学硕士学位论文,2004.
郭世宁,张克家,于建辉.黄芪多糖的药理研究[J].中兽医学杂志,1996,83(2)：41-43.
高微微,曹丽,李展等.黄芪成分对鸡外周血淋巴细胞增殖反应的影响[J].中国兽医杂志,2001,35(6)：16-18.
何彦丽,苏俊芳.中药多糖抗肿瘤免疫药理研究的新思路[J].中国中西医结合杂志,2003,23(1)：73-76.
侯慧英,秦荣,王玉珍.蒙药广枣总黄酮对小鼠体液免疫力影响的研究[J].中国民族医药杂志,1998,4(4)：38-39.
侯玉慧,杜爱芳,胡松华.复方黄芪对雏鸡免疫功能的增强作用.中国兽医杂志,2006,42(3)：31-32.
胡松华,杜爱芳等.复方黄芪、绞股蓝对鸡新城疫疫苗免疫效果的影响[J].中兽医学杂志,2000,2(99)：1-3.
胡庭俊,梁纪兰,程富胜等.黄芪多糖对鸡脾脏淋巴细胞内游离钙离子浓度的影响[J].中兽医医药杂志,2001,(5)：4-6.
胡元亮,陈文喜.中药免疫药理学研究近展[J].中国免疫学杂志,1997,13(专刊)：96-98.
胡元亮,孔祥峰,李祥瑞等.10种中药成分对CEF的增殖和抵抗NDV感染的影响.畜牧与兽医学报,2004,35(3)：301-305.
胡元亮,周新民,陈玉库等.芪蓝抗毒饮预防鸡法式囊病和增强免疫作用的实验研究.畜牧与兽医,2003(7)：3-5.
胡斌杰,陈金锋,王宫南.超声波法与传统热水法提取灵芝多糖的比较研究.食品工业科技,2007,28(2)：190-192.
贺正全,王宝琴.蒙古黄芪化学成分的分离鉴定[J].药学学报,1990,25(9)：694-698.
贺生中,许发芝,周建强等.黄芪多糖对犬淋巴细胞转化影响的实验研究[J].动物科学与动物医学,2004,21(11)：58-60.
贺玉琢.十全大补汤对肠道免疫系统生成细胞因子的调节作用[J].国外医学中医中药分册,1999,21(3)：57-58.
花象柏,王兴金,何后军等.黄芪对I系苗接种鸡血液生化指标与免疫功能影响的研究[J].中兽医学杂志,1993,(2)：11-14.
焦艳,闻杰,于晓红等.膜荚黄芪颈叶总黄酮对小鼠细胞免疫功能的影响[J].中国中西医结核杂志,1999,19(6)：356-357.
金春英,林金清,李雯君.超临界流体萃取技术在中药提取中的应用与发展趋势.化学工程与装备,2006,(5)：40-42.
吉荣,崔治中,王锡乐等.分子克隆化禽网状内皮组织增生症病毒传染性及其前病毒全基因组序列研究[J].病毒学报,2005,21(6)：448-455.
孔祥峰,胡元亮,李祥瑞等.9种中药成分对新城疫Ⅳ系疫苗免疫雏鸡血液中血凝抑制抗体水平的影响[J].畜牧兽医学报,2004,35(4)：468-472.
雷红,祁成年.中草药作为免疫增强剂的作用.塔里木农垦大学学报,1999,9(11)：36-37.
雷连成,韩文瑜.免疫增强剂的研究进展[J].中国兽药杂志,2002,36(6)：36-38.雷载权.中药学.上海科学技术出版社,1995.
骆延波,刘万成,张再辉等.复方免疫活性因子与左旋咪唑对雏体液免疫应答影响的研究.养禽与禽病防治,2004,(3)：11-12.
李繁,谢明权,林辉环等.免疫增强剂促进鸡球虫细胞免疫的研究[J].中国预防兽医学报,2000,22(3)：196-199.
李宏全,段县平,马海利等.黄芪多糖对鸡新城疫和传染性腔上囊病疫苗免疫力的影响[J].中国兽医科技,2001,31(9)：12-14.
李宏全,段县平,马海利等.黄芪多糖提高鸡抗氧化作用对免疫功能的影响[J].山西农业大学学报,2001,22(1)：78-81.
李萍,何明,黄起壬.油茶皂苷对缺氧复氧所致大鼠心脏损伤的保护作用及其机制探讨.中草药,2000,31(11)：841-844.
李淑慧,楚延,李晓辉.三七总皂苷抗炎作用机制的实验研究.中草药,2000,30(9)：676-677.
李文魁,林新,彭勇等.远志属药用植物皂苷类成分及其药理活性.1998,13(2)：60-64.
李文魁,肖培根,张如意等.朝鲜淫羊藿的化学成分(Ⅰ)[J].天然产物研究与开发,1994,6(4)：12-18.
李文魁,肖培根,潘景歧等.朝鲜淫羊藿的化学成分(Ⅱ)[J].中国药学杂志,1995,30(8):455-456.
李文魁,肖培根,廖矛川等.朝藿苷E的分离和结构[J].高等学校化学学报,1995,16(12)：1892-1895.
李秀真,朱万孚.新型疫苗佐剂-细菌CpG寡核苷酸[J].国外医学预防诊断治疗用生物制品分册,2004,27(3)：97-101.
李平.碱提法茱萸多糖的理化性质及抗氧化活性研究.中草药,2003,34(11)：973-976.
李宏燕,樊君.大枣多糖的水提醇沉工艺研究.宁夏工程技术,2005,4(3)：265-267.
李义,吴婷婷,谭夕东.左旋咪唑对中华绒鳌蟹非特异性免疫功能及抗病力的影响.水产学报,2007,31(6)：785-787.
李永泉,吴炬,花立明等.白阿魏菇菌丝体多糖分离纯化工艺的优化和结构分析[J].兰州大学学报(自然科学版),2003,39(4)：50-54.
李艳华,田国彬,李雁冰等.某复方中药抗禽流感病毒的免疫抑制[J].中国预防兽医学报,2004,26(6)：416-419.
李子石,金晓红,宋振清等.中草药饲料添加剂在养鸡业中的应用现状[J].现代畜牧兽医,2006(5)：6-8.
李小定,欧阳天贽,荣建华等.灰树花多糖PGF-2的理化性质及化学结构的初步表征[J].菌物学报,2005,24(2)：245-250.
李义,吴婷婷,谭夕东.左旋咪唑对中华绒鳌蟹非特异性免疫功能及抗病力的影响.水产学报,2007,31(6)：785-791.
刘玉堂,巴图.左旋咪唑对新城疫免疫效果的影响[J].中国兽医杂志,1992,18(1)：33.
刘志伟,姜华年.菱叶红景天多糖的提取、纯化鉴定及理化特性研究[J].食品科学,2005,26(3)：60-63.
刘勤,刘永隆.黄甘草化学成分的研究[J].药学学报,1989,24(7)：525.
刘青梅,杨性民,邓红霞等.紫菜多糖提取分离及纯化技术研究[J].浙江大学学报,2005,31(3)：293-297.
刘家国,胡元亮,张宝康等.淫羊藿-蜂胶佐剂的免疫调节作用[J].中国兽医学报,2000,20(4)：383-385.
刘家国,胡元亮,张宝康等.淫羊藿-蜂胶佐剂对小白鼠NK和腹腔巨噬细胞活性的影响[J].中国中国免疫学杂志,1996,12(增刊)：251-253.
刘铭佩.高乌甲素对炎症性疼痛的镇痛作用.中国药物与临床,2003,3(3)：244-245.
刘永杰,李庆章,郝艳红.香菇多糖和黄芪多糖对马立克氏病强毒感染雏鸡白细胞介素2活性和淋巴细胞增殖反应的影响[J].中国兽医杂志,1999,25(7)：3-5.
梁荣,郭抗抗,伊岚等.中药免疫增强剂提高鸡免疫功能的研究[J].中国兽医科技,1998,28(9)：11-12.
梁中琴,王晓霞,顾振纶等.云芝糖肽诱导人外周血淋巴细胞产生IL-6的研究[J].中国药学杂志,1999,34(10)：700-702.
廉宜君.中药复方免疫增强剂中多糖的分离纯化与免疫活性研究.硕士研究生论文.2007,28-29.
黎海彬,李琳,郭远等.药用植物有效成分提取技术现代化工.2002,22(5)：59-62.
楼兰花,沃兴德.参芦总皂甙通过cAMP、cGMP对人淋巴细胞E-玫瑰花结形成的影响[J].中国免疫学杂志,1986,2(1)：30-33.
吕凤林,何凤慈.利用纳米材料做多肽疫苗佐剂的思考[J].生物化学与生物物理进展,2001,28(6)：832-835.
吕殿红,王刚,张毓金.复方免疫增强剂对鸡新城疫和传染性腔上囊病疫苗免疫效果的影响[J].中国兽医科技,2000,30(10)：21-22.
吕曙华,朱永智,吴寿金.内蒙黄芪地上部分黄酮成分的研究[J].中草药,1990,21(6)：9-10.
马学文,毛志明,高建英等.免疫增强剂提高破伤风免疫马血浆效价试验研究[J].甘肃畜牧兽医,2005,181(2)：8-9.
孟宪荣,李庆章,曲琪环等.黄芪多糖和香菇多糖对马立克氏病强毒感染雏鸡巨噬细胞活性和白细胞介素-1体外诱生活性的影响[J].中国兽医杂志,2002,38(7)：33-34.
马莉,唐健元,李祖伦等.板蓝根多糖分离纯化及其性质的研究.中草药,2007,38(8)：1143-1146.
孟庆勇,刘志辉,徐美奕等.半叶马尾藻多糖的提取和分析[J].光谱学与光谱分析,2004,24(12)：1560-1562.
聂中越,程淑娟.中药在未来抗感染治疗中开发创新的探索[J].深圳中西医杂志,1998(6)：40-42.
钮明洋,姚文兵,姚金凤等.超滤法在海洋真菌多糖分离纯化工艺中的应用.中国生化药物杂志,2004,25(3)：161-162.
邱吕庆.鸡新城疫-减蛋综合征蜂胶佐剂灭活二联苗的研究[J].中国兽医科技,1996,26(12)：2-5.
屈洪岩,郭玉璞,陆钢等.花粉多糖对新城疫弱毒苗效果的影响[J].中国兽医杂志,1998,9： 15-17.
乔善义,王立岩,赵毅民等.山药多糖的提取分离和结构测定[J].中国天然药物,2003,1(3)： 155-156.
秦竹丽,江元汝.超临界萃取技术在生物碱提取中的应用进展.化工时刊,2006,20(7)： 60-63.
沈志强.禽霍乱蜂胶灭活疫苗研究与应用总结[J].中国畜禽传染病,1995,(4)：27-30.
石玉祥,李存等.中药对新城疫灭活苗免疫效果的观察[J].中国兽医杂志,2000,10:18-19.
睢大员,陈满秋,于晓凤等.人参Rb组皂苷对犬实验性心肌梗死的保护作用.中草药,2001,32(2)：136-139.
孙月平,王金宝,张乐萃等.黄芪多糖对免疫抑制鸡某些免疫指标影响的研究[J].莱阳农学院学报,1999,16(1)：61-63.
宋立人,洪恂,丁绪亮等.现代中药学大辞典(下册)[M].北京：人民卫生出版社,2001.
孙朋悦,文晔,徐颖等.朝鲜淫羊藿的化学成分[J].药学学报,1998,33(12)：919-922.
孙朋悦,徐颖,文晔等.朝鲜淫羊藿的化学成分Ⅰ[J].中国药物化学杂志,1998,8(2)：122-125.
谭宫屏,李东真,许静亚等.雷公藤根、茎、叶、花的免疫药理作用比较[J].中药材,1989,12(3)：38-41.
唐希灿,郭新,冯洁.单胺类递质对3-乙酰乌头碱镇痛作用的影响.中国药理学报,1988,9(3)：216-220.
唐雪明,胡元亮,张宝康等.应用乳酸脱氢酶释放法研究黄芪多糖耐雏鸡外周血中自然杀伤细胞活性的影响[J].中兽医学杂志,1997,20(2)：9-11.
剡根强,谷新利,王玮等.中药免疫增强剂对雏鸡免疫功能的影响[J].中国兽医科技,1998,28(2)：31-32.
邰光远,李荣仙,孙世海.中药增强雏鸡免疫力实验[J].中兽医医学杂志,1998,1：5-7.
屠鹏飞,郭洪祝,果德安.中药与天然药物活性成分研究及新药的发现[J].北京大学学报,2002,34(5)：513-518.
唐雪明,胡元亮,张宝康.黄芪多糖对雏鸡外周血淋巴细胞转化功能的影响[J].中国兽医学报,1998,18(3)：3-5.
汪东风.茶多糖的组分及理化性质[J].茶叶科学,1996,16(1)：1-8.
吴长德,吕戴,方永辉.复方女贞子-黄芪制剂对雏鸡的免疫增强作用.黑龙江畜牧兽医.2003年第1期：30-31.
吴寿金,张晓闻.近年来皂苷药理活性的研究概况.国外医药.植物药分册1994,9(6)： 246-252.
吴寿金,赵泰,秦永琪.现代中草药成分化学(第一版).北京：中国医药科技出版社,2002,324-330.
王长云,管华诗.多糖抗病毒作用研究进展Ⅰ：多糖抗病毒作用[J].生物工程进展,2000,20(1)：17-20.
王超英,柳纪省,白银海等.鸡用天然免疫增强剂的研究：天然免疫增效药物的筛选[J].中国兽医科技,1998,28(7)：13-15.
王德云,胡元亮,孔祥峰等.中药成分对培养的鸡脾脏淋巴细胞增殖的影响[J].畜牧兽医学报,2005,36(11),1210-1214.
王福传,方昌阁,张玉换等.复方中草药免疫增强剂对蛋鸡免疫效果的研究[J].畜牧兽医学报,2006,37(2)：187-192.
王福传,韩一超,赵洪恩等.复方中草药免疫增强剂对鸡免疫器官组织形态学影响的研究[J].中国预防兽医学报,2001,23(6)419-421.
王福传,韩一超,张玉换等.兽用中草药高效免疫增强剂研究初报[J].山西农业科学,2001,29(2)：74-80.
王弘,吴梧桐,顾学袭.脂质体作为生物大分子载体的研究进展[J].药物生物技术,2002,9(3)：171-174.
王惠燕.党参多糖对小鼠细胞免疫的调节作用[J].中国药理学通讯,1989,5(6)：276-28.
王建文.中草药免疫作用概述[J].中兽医医药杂志,1991,(6)：19-21.
王金华.小柴胡汤和干扰素α/β对小鼠肺组织肿瘤坏死因子α和白细胞介素-1β生成的增加作用[J].国外医学中医中药分册,1996,18(6)：39.
王立人,梁智辉,冯玮.黄芪、刺五加合剂对免疫功能调节的研究[J].中国实验临床免疫学杂志.199'7,10(4)：61-66.
王世英,张登荣,杜方良等.中药对雏鸡感染IBDV前后法氏囊相对重量及血清γ-球蛋白动态变化的影响[J].中兽医医药杂志,1993, (4)：3-5.
王义,朱明艳,任洪志等.目前我国畜禽传染病防制存在的问题[J].畜牧兽医科技信息,2004,6：44.
王涌,胡志前,成志刚等.中药黄芪对阻塞性黄疸大鼠细胞免疫功能的影响[J].肝胆外科杂志,2000,8(1)：64-66.
王超英,柳纪省,白银海等.鸡用天然免疫增强剂的研究：天然免疫增效药物的筛选[J].中国兽医科技,1998,28(7)：13-15.
王德云,胡元亮,孔祥峰等.中药成分对雏鸡外周血T淋巴细胞转化的影响[J].中国兽医学报,2004,24(6)：578-580.
王德云,胡元亮,张宝康等.几种中药成分与IL 2免疫协同作用的比较.南京农业大学学报,2005,28(3)：140-142.
邬方宁.超声提取技术在现代中药中的应用.中草药,2007,38(2)：315-316.
王惠康,何侃,叶嘉麟.内蒙黄芪化学成分的研究(Ⅰ)[J].中草药,1989,20(5)： 6-8.
王建军.径直黄芪化学成分研究及其提取物体外抑菌活性试验.硕士研究生论文,2007,49-56.
王金宝,孙月平,张乐翠等.中药对免疫抑制鸡某些细胞免疫指标影响的研究.中兽医医药杂志,1999,3：9-11.
王顺春,何巍,方积年.徐长卿中一种新葡聚糖化学结构的研究[J].生物化学与生物物理学报,2000,32(4)：369-372.
王卫国,赵永亮,韩山宝等.香菇多糖分离纯化技术研究.中国食用菌,2002,21(2)：30-32.
王樟,许时婴.不添加蛋白酶从百合根茎中提取分离纯化水溶性非淀粉多糖.浙江大学学报：农业与生命科学版,2004,30(2)：159-163.
韦巍.多糖的研究进展.国外医学.药学分册,2005,32(3)：179-184.
伍义行,王芳.中药的双向免疫调节作用研究现状.中兽医医药杂志,1999,4,19-22.
夏尔宁.猴头菇多糖的提取分离、理化分析给生物活性的初步研究[J].南京药学院学报,1986,2：129.
夏英,李家琦,黄健.多糖免疫作用的试验研究[J].上海中医药杂志,1994,11：42-43.
肖崇厚,杨松松,洪筱坤等.中药化学(第四版).上海科学技术出版社,2000,265-274.
谢庆阁.畜禽重大疫病免疫防制研究进展[M].中国农业科技出版社,1995：143-168.
徐杰诚,张明珠.胞壁酰二肽(MDP)及其类似物的合成[J].化学学报,1995,53：509-515.
徐诺.黄芪的药性：对体外抗体生成作用的影响[J].国外医学中医中药分册,1998,20(2)： 40.
夏春英,陈力,孟羽贤.黄芪注射液对慢性肾炎患者红细胞免疫功能及IL-6的影响.中国中医药科技,1997,4(5)：263.
徐任生主编.天然产物化学(第二版)[M].北京：科学出版社,2004.
夏英,李家琦,阎佩君等.黄茂多糖(APS)免疫作用的实验研究.上海中医药杂质,1994,05(11)：42-43.
夏永高,葛彪,蒋伟娟等.地巴唑、左旋咪唑对雏鸡新城疫疫苗和巴氏杆菌灭活苗免疫应答的影响.上海交通大学学报(农业科学版).2003,21：90-92.
杨风华,康成,李淑华等.黄芪水溶性黄酮类对小鼠细胞免疫功能的影响[J].时珍国医国药,2002,(12)：718-719.
杨金明.黄芪对S180荷瘤小鼠的免疫增强及抑癌作用[J].中国实验临床免疫学杂志.1995；7(1)：41.
杨希国,张秀英.中草药对畜禽免疫药理作用研究进展.中兽医医药杂志,2004,1(124)：16-18.
于飞,康金凤,李巍.左旋咪唑对细粒棘球蚴感染小鼠不同时期T细胞亚群的影响.中国病原生物学杂志,2007,22(1)：57-59.
杨焕荣,杨夏,余晓玲等.黄芪对慢性肾功能衰竭患者免疫功能的影响.中西医结合实用临床急救,1997,4(9)：404-405.
严金龙,吕志敏.植物多糖的提取分离和应用市场.化工科技市场,2004,27(11)：41-44.
袁龙刚,何云,刘林丽.现代提取分离技术在食药用菌多糖分离纯化中的应用.生物技术通报,2006,(5)：76-79.
姚新生,吴立军,吴继洲等.天然药物化学.第4版.北京：人民卫生出版社,2006：102.
姚新生.天然药物化学(第二版).北京人民卫生出版社,1995：360-362.
杨云,苗明三,王浴铭.怀地黄多糖化学研究[J].时珍国医国药,1999,10(8)：564-565.
郁利平.大豆对小鼠细胞免疫功能的增强作用[J].中国免疫学杂志,1992,8(3)：191.
曾陇梅.海洋天然产物研究新进展.有机化学,1989,(9)：402-413.
张春杰,程相朝,李银聚等.中药免疫增强剂对HVT免疫雏鸡免疫器官中免疫活性细胞的影响[J].陕西农业大学学报,2002,22(4)：360-361.
张昌军,原方圆,邵红兵.超声波法在提取多糖类化合物中的应用研究.化工时刊,2007,21(2)：54-56.
张华芳,李士敏.降血糖天然药物研究进展.中草药,2000,31(增刊)：184-186.
张惠灵,王翔毅,杨玉梅等.冉乌头碱的镇痛作用及其对麻醉大鼠心电图的影响.包头医学院学报,2000,16(4)：261-264.
张乐萃,王金宝,孙月平等.复方中药多糖对鸡免疫器官形态学的影响[J].中国兽医科技,1998,28(8)：26-28.
张乐萃,王金宝,孙月平等.新城疫中药多睹佐制油乳苗对鸸免疫器官投育的影响.中国兽医学报,1998；15(4)：375-351.
张建新.黄芪对小鼠免疫功能影响的研究[J].时珍国医国药,2000,11(6)：488.
张林维,赵帜平,沈业寿.当归多糖的分离纯化及其部分性质的研究[J].生物学杂志,1998,15(3)：12-14.
张如意.朝鲜淫羊藿的化学成分[J].天然产物研究与开发,1994,6(3)：4-8.
张如意,张建华,汪茂田.乌拉尔甘草中皂甙的研究[J].药学学报,1986,21(7)：510.
张倩,江萍,秦礼康等.多糖功能的研究进展[J].贵州农业科学,1998,26(4)：59-60.
张庆茹.中草药免疫促进作用的研究进展[J].中兽医医药杂志,1997,5：15-16.
张书霞,陈万芳,宋大鲁.淫羊藿-蜂胶合剂对鸡马立克氏病的影响[J].南京农业大学学报,1996,19(1)：73-76.
张述斌,薛掌林,刘瑞生等.黄芪多糖、淫羊藿多糖对鸡新城疫疫苗免疫增强作用的研究[J].甘肃畜牧兽医,2004,(4)：24-25.
张廷伟.头花千金藤提取剂千金藤碱对干扰素的诱导作用及与其他药物的比较研究[J].国外医学中医中药分册,1998,20(4)：37.
张文华,俞富荣,尤丽芬等.黄芪甙对霍乱弧菌LPS体外诱生TNF的影响[J].上海免疫学杂志,1996,16(4)：224-226.
张兴权,陈鸿珊.柴胡多糖的免疫药理作用[J].中国药理学与毒理学杂志,1989,3(1)：30-33.
张炎,张咏南.胞壁酰二肽衍生物增强汉坦病毒灭活疫苗免疫原性的佐剂活性[J].国外医学.预防、诊断、治疗用生物制品分册,1998,21(5)：233-234.
张有为,窦德强,陈英杰等.人参皂苷对人体骨肉瘤细胞U20S增殖的影响.中草药,2001,32(3)：232-233.
张悦,戚晓红,冯振卿等.免疫佐剂的研究进展[J].中国血吸虫病防治杂志,2004,16 (3)：236-238.
张晓静,刘会东.植物多糖提取分离及药理作用的研究进展.时珍国医国药,2003,14(8)：495-496.
周洪波,操继跃等.中草药免疫促进剂的药效学研究进展[J].中国兽药杂志,2002,36(6)：42-44.
周淑英,卢振初.黄芪多糖对重组白介素2激活LAK细胞活性的调节作用[J].南京中医学院学报,1994,10(6)：29-31.
周义乾,刘玉林,刘清珍等.黄芪注射液对肺结核患者T细胞亚群的影响及近期疗敏[J].第四军医大学学报,1999,20(10)：921-923.
朱恩园,贺庆,王峥涛等.党参化学研究[J].中国药科大学学报,2001,32(2)：94-95.
庄茂辛,吴耀生,李曼玲等.黄芪、党参、人参多糖对豚鼠免疫功能的影响[J].中国药学杂志,1992,(11)：653.
庄国庆,孙淑红,崔治中,曲立新.鸡马立克氏病毒和网状内皮增生病病毒共感染商品代肉鸡时的相互作用.中国病毒学,2006,21(2)：157-162.
郑虎占,董泽宏,余靖等.中药现代研究与应用[M].北京：学苑出版社,1997,1256-1279.
郑宝,方中,舒启松等.复方黄芪饮(囊炎康)防治IBD试验[J].中国兽药杂志,1998,32(3)： 53-55.
赵克胜,丁丽轩,孔海燕等.黄芪多糖增强人外周血单个细胞产生肿瘤坏死因子的研究[J].中国中西医结合杂志,1993,(13)：263.
赵永芳.生物化学技术原理与应用[M].武汉：武汉大学出版社,1994.
赵国华,陈宗道,李志孝等.活性多糖的研究进展[J].食品与发酵工业,2001,27(7)：45-48.
赵银丽,李国喜,李慧勤.黄芪对固始鸡免疫性能的影响[J].河南农业科学,2004,(11)：90-91.
Ameri A., Gleitz J., Petera T..Aconitine inhibits epileptiform activity in rat hippocampal slices. Naunyn Schmiedebergs Arch Pharmacol,1996,354(1):80-85.
Ameri A., Metzmeier P., Peters T.. Frequency-dependent inhibition of neuronal activity by lappaconitine in normal and epileptic hippocampal slices. Br. J. Pharmacol.,1996,118(3): 577-584.
Ameri A., Seitz U.. Effects of mesaconitine on [3H] noradrenaline uptake and excitablity in rat hippocampus. Exp. Brain Res.,1998,121(4):451-456.
Ameri A., Simmet T.. Antagonism of the aconitine-induced inexcitability by the structurally related aconitum alkaloids, lappaconitine and ajacine. Brain Research,1999,842:332-341.
Ameri A.. The effects of Aconitum alkaloids on the central nervous system. Neurobiol,1998, 56(2):211-235.
Ameri A..Inhibition of stimulus-triggered and spontaneous epileptiform activity in rat hippocampal slices by the Aconitum alkaloid mesaconitine. Eur. J. P hrmacol.,1998,342(2-3): 183-191.
Bradney C P, Se mpowski G D, Liao H X, et al. Cytocines as adjuvants for the induetion of Anti-immunodeficieney virus peptide immunoglobulin G and IgA antibodies in serum and mucosal secretions after nasal immunization[J]. J Virol,2002,76:517.
Babiuk L A, Misra V.. Levamisole and bovine immunity:in vitro and in vivo effects on immune responses to herpesvirus immunization[J]. Can J Microbiol 1981,27:1312-1319.
C.L.Cooper, H.L.Dvais, M.L.Morris, et al.Safety and immunogenicity of CpG ODN injection as an adjuvant to Fluarix influenza[J].vaccine,2004,23-24:3136-3143.
Chen H L, Li D F, Chang B Y, et al. Efects of Chinese herbal polysaccharides on the immunity and growth performance of young broilers[J]. Poult Sci,2003,82(3):364-370.
Chow Y H, Chiang B L, Lee Y L, et al.Development of Thl and Th2 populations and the nature of immune responses to hepatitis B virus DNA vaccine can be modulated by codelivery of various cytokine genes[J].J Immunol,1998,160(3):1320-1329.
Chow Y H, Chiang B L, Lee Y L, et al.Development of Thl and Th2 populations and the nature of immune responses to hepatitis B virus DNA vaccine can be modulated by codelivery of various cytokine genes [J].J Immunol,1998,160(3):1320-1329.
Chow Y H, Hunag W L, Cli W K,et al.Improvement of hepatitis B virus DNA vaccines by plasmids coexpressing hepatitis B surface antigen and interleukin-2[J].J Virol,1997,71 (1): 169-178.
Chu RS, McCool T, Greenspan NS, et al.CpG oligdeoxy nucleotides act as adjuvants fox Pneum ococcal Polysaccharide-Protein conjugate vaccines and enhance antipolysac charide immunoglubulin G 2a (IgG2a) and IgG3 antibodies[J].Inefet Immun,2000,68:1450-1456.
Commenges D., Scotet V., Renaud S.,et al.. Intake of flavonoids and risk of dementia. Eur. J. Epidemiol.,2000,16:357.
Cooper P D., McComb C., Steele E J..The adjuvanticity of algammulin, a new vaccine adjuvant[J],1991,9 (6):408-415.
Cos P., Calomme M., Sindambiwe J.B.,et al. Cytotoxicity and lipid peroxidation-inhibiting activity of flavon oids. Planta Med.,2001,67:515.
Cos P., Ying L., Calomme M., et al.. Stucture-activity relationship and classification of flavonoids as inhi bitors of xanthine oxidase and superoxide scavengers. J. Nat. Prod.,1998, 61:71.
Di Carlo G., Mascolo N, Izzo A. A, et al..Flavonoids:old and new aspects of a class of natural therapeutic drugs.Life Sci.,1999,65:337.
Dixon T.A., Steele C.L.. Flavonoids and isoflavonoids-A gold mine for metabolic engineering. Trends plant Sci.,1999,4:394-400.
Dzhakhangirov F.N.,Sultankhodzhaev M.N.,Tashkhodzhaev B.,et al..Diterpenoid alkaloids as a new class of antiarrhythmic agents.Structure-activity relationship.Chemistry of Natural Compounds,1997,33(2):190-202.
Elson CO, Ealding W. Generalized systemic and mucosal immunity in mice after mucosal Stimulation with cholera toxin[J].J Immunol,1984,132:2736.
Ferrali M., Signorini C., Caciotti B., et al.. Protection against oxidative damage of erythrocyte membrane by the flavonoid quercetin and its r elation to iron chelating activity. FEBS Lett.,1997,416:123.
Gallichan W S, W oollstencroft R N, Guarasci T, et al. Intranasal immunization with CpG oligodeoxynucleotides as an adjuvant dramatically increases IgA and Protection against herpes simplex virus-2 in the genital tract[J].J Immunol,2001,166:3451-3457.
Geng Chang-shan. Enhancing effect of Lycium barbarum polysaccharides on the interleukin-2 activity in mice.1989,3:18-22.
Gherardi M M, Ra mirez J C, Estebah M.Towards a new generation of vaccines:the cytokine IL-12 as an adjuvant to enhance cellular immune responses to pathogens during prime-booster vaccination regimens[J].Histol Histopathol,2001,16(2):655-667.
Goyal K, Human L.Gene therapy using DC Chol cationic liposomes[J].Liposome Research, 1994,4:1075.
Guo X., Tang X.C.. Lappaconitine and N-deacetyllapaconitine potentiate footshock-induced analgesia in rats. Life Sci.,1991,48(14):1365-1370.
Gupta RK.Aluminum compounds as vaccine adjuvnats.Adv Drug Deliv Rev.1998,32 (3):155-172.
Gutser U.T., Friese J., Heubach J.F., Matthiesen T., Selve N., Wiffert B., Gleitz J..Mode of antinociceptive and toxic action of alkaloids of Aconitum species. Naunyn Schmiedebergs Arcg. Phrmacol.,1998,357(1):39-48.
Harper EG,Simpson EL,Takiguchi RH,et al. Efalizumab therapy for atopic dermatitis causes marked increases in circulating effector memory CD4+T cells that express cutaneous lymphocyte antigen[J].J Invest Dermatol,2008,128(5):1173.
Harrod T, Martin M, Russell M W. Long-term persistence and recall of immune response in aged Mice after mucosal immunization[J]. Oral Micro boil Immunol,2001,16(3):170.
Hasegawa H., Uchiyama M.. Antimetastatic efficacy of orally administered ginsenoside Rbl in dependent on intestinal bacterial hydrolyzing potential and significance of treatment with an active bacterial metabolite. Planta. Med.,1998,64:696-700.
Heubach J.F.,Schule A..Cardiac effects of lappaconitine and N-deacetyllappaconitine, two diterpenoid alkaloids from plants of the Aconitum and Delphinium species. Planta Medica., 1998,64:22-26.
Hikino H., Takata H., Fujiwara M., et al.. Mechanism of inhibitoryaction of mesaconitine in acute inflammations. Eur. J. Phrmacol.,1982,82(1-2):65-71.
Hollman P.C.H.. Evidence for health benfits of plant phenols:local or systemic effects. J. Sci. Food Agric.,2001,81:842.
Hoshino J, Naganu ma F, Nagai R. Ventricular fibrillation triggered by a ruptured sinus of Valsalva aneurysm[J]. Heart,1998,80(2):203-204.
Hassan AB. Atta AH, Sdiman RT. et al. Effect of levamisole hydrpchoricle on the immune response to Newcastle disease virus vaccine in chickens[J]. Indian Vet J,1989,66:389-394.
Hogarth-Scott RS, Liardet DM, Morris P J. Levamisole vaccine combination.1. Heightened antibody response[J]. Aust Vet J 1980,56:285-291.
Jin R. Effects of Chinese herbson macrophage functions in N-butyl-N-butanolniatrosoamine treated mice[J].Imunopharmacology and Immunotoxicology,1996,18:105.
KONG X,HU Y,RUIR,et al. Effects of Chinese herbal me-dicinal ingredients on peripheral lymphocyte prolifer-ation and serum antibody titer aftervaccination in chicken[J]. Int Immu-nopharmaca,2004,4(7):975-982.
Kunari J, Sahoo P K. Dietary levamisole modulates the immune response and disease resistance of Asian catfish Clarias batrachus(Linnaeus). Aquaculture Research.2006,37: 500-508.
Kim J J, Yang J S, Manson K H.et al.Modulation of antigen 7 specific cellular immune responses to DNA vaccine in rhesusmacaques through the use of IL-2, I FN-gamma, or IL-4 gene adjuvant[J].Vaccine,2001,19(17-19):2496-2505.
Kimura.Dominant constituent responsible for the neuromuscular blocking action of the Iapanese-sino medicine "bushi" (aconite root). Jpn. J. Pharmacol.,1988,48(2):290-293.
Knekt P., Jarvinen R., Reunanen A., et al.. flavonoid intake and coronary mortality in Finland: A cohort study., Br. Med. J.,1996,312:478-481.
Lingh, M A, Gelfand E W. Increaded cyclic adenosine mono-phosphate levels bolck progression but not initiation of human T cell proliferation[J]. J Immunol,1990(145):449-455.
Lowe R J. Levamisole as an immunostimulant[J]. Vet Rec 1980,106:390.
Le Marchand L.. Cancer preventive effects of flavonoids. Biomed. Pharmacother,2002,56: 296.
Lee Y L, Tao M H, Chow Y H, et al. Construction of vectors expressing bioactive heterodimeric and single-chain murine interleukin-12 for gene therapy[J]. Hum GeneTher, 1998,9(4):457-465.
Marinaro M, Staars H F, Hiroi T, et al. Mucosal adjuvant effect of cholera toxin in mice results from induction of The T helper (Th2) cells and IL-4[J]. J Immune,1995,155:4621.
Mojzisova G., Kuchta M.. Dietary flavonoids and risk of coronary heart disease. Physiol. Res.,2001,50:529.
Morand C., Crespy V., Manach C., et al.. Remesy C.. Plasma metabolites of quercetin and their antioxident properties. Am. J. Physiol.,1998,275:R212-R219.
Murayama M., Hikino H.J.. Stimulating actions on ribonucleic acid biosynthesis of aconitines, diterpenic alkaloids of Aconitum roots. Ethnopharmacol.,1984,12(1):25-33.
Maaten, Schmerr Sacks. Levamisole does not affect the virological and serological responses of bovine leukemia virus-infected cattle and sheep[J]. Can J Comp Med 1983,47:474-479.
Marnoch R, DiesadyL L.. Production of mustard protein isolates from orientalmustard seed (Brass-ica junceaL). Journal of the American Oil Chemists Society,2006,83(1):5-7.
Nicklas W. Aluminum salts[J]. Res Immunol,1992,143(5):489-494.
Nijveldt T.J., Van Nood E., Van Hoorn D.E.C., et al.. flavonoids:a review of probable mechanisms of actions and potential applications. Am. J. Clin. Nutr.,2001,74:418.
Oh M., Choi Y.H., Choi S.. Anti-proliferating effects of ginsenoside-Rh2 on MCF-7 human breast cancer cells. Int. J. Oncol.,1999,14:869-875.
Ohnon, Miurat, Miuran. Structure and biology activities of hypochlorite oxidized zymosan [J], Carbohydratepolymers,2001,44:339-349.
Ono M., Satoh T.. Pharmacological studies of lappaconitine:possible interaction with endogenous noradrenergic and serotonergic pathways to induce antinociception. Jpn. J.Pharmacol.,1992,58(3):251-257.
Pertmer T M, Oran AE, Madorin CA.et al. Thl genetic adjuvants modulate immune responses in neonates[J]. Vaccine,2001,19(13-14):1764-1771.
Rice-Evans C.A., Miller N.J.. Antioxidant activities of flavonoids as bioactive components of food. Biochem. Soc. Trans.,1996,24:790-795.
Robert S, Northrup, Anthony S, et al. Adjuvant effct of cholera enterotoxin on the immune Response of the mouse to sheep red blood cells[J]. J Infect Dis,1972,125(6):672.
Pratt D.E.. Natural antioxidants from plant materia. ACS Symp. Ser. No.5071992:54-71.
Palanskii. A M, Ryagin S T and German V V. Enhancement of the efficacy of vaccination of broiler chicks against Newcastle disease by means of levamisole and sodium nucleinate[J]. Vet Bull,1991,61(2):162.
Panigrahy B, Grumbles LC, Millar D., et al..Antibiotic-induced immunosuppression and levamisole-induced immunopotentiation in turkeys[J]. Avian Dis 1979,23:401-408.
Symoens J, Rosenthal M. Levamisole in the modulation of the immune response:the current experimental and clinical state[J]. J Reticuloendothel Soc 1977,21:175-221.
Seitz U., Ameri A.. Different effects on [3H] noradrenaline uptake of the aconitum alkaloids aconitine,3-acetylaconitine, lappaconitine, and N-desacetyllappaconitine in rat hippocampus. Biochem. Pharmacol.,1998,55(6):883-888.
Song K, Chang Y, Prudhomme G J. Regulation of T-helped activity and enhancement of tumor immunity by combined DNA based vaccination and nonviral cytokine gene transfer[J]. Gene Ther,2000,7(6):481-492.
Stede Y V, Verdonck F, Vancaenehhe m S, et al. Cp G-oligonu-cleotides as an effective adjuvant in pags fox intromuscular immunizations[J].Vet Im mu and Immunopath,2002, 86(1):31-41.
Vyas Vyas GP, Dholakia PM Kathuria LG. Studies on immunomodalation by levamisole along with vaccination in chicks against Ranikhet Disease[J]. Indian Vet J,1987,64: 456-462.
Wissing KM, Fomegne G, Broeders N, et al. HLA mismatches remain risk factors for acute kidney allograft rejection in patien ts receiving quadruple immunosuppression with anti-interleukin-2 receptor antibodies[J]. Transplantation,2008,85(3):411.
Wright S.N., Irreversible block of human heart (hHl) sodium channels by the plant alkaloid lappaconitine. Mol. Pharmacol.,2001,59(2):183-192.
Xin K Q, Hamajima K, Sasar S, et al. Intranasal administration of human immunodeficiency virus type-1(HIV1)DNA vaccine with interleukin-2 expression plasmid enhances cell-memated immunity against HlV-1[J]. Immunology,1998,94(3):438-444.
Yamanaka H., Doi A., Ishibashi H., Akaike N.. Aconitine facilitates spontaneous transmitter release at rat ventromedial hypothalamic neurons. Br. J. Pharmacol.,2002,135(3):816-822.
Zhang W J. Biochemical Study Technology of Complex Polysaccharides[M]. Shanghai:Shanghai Science and Technology Publishers,1987.
Zhu Y Zh, Lu Sh H, Yoshihito O, et al. Two new cycloartane-type glucosides, mongholicoside Ⅰ and Ⅱ, from the aerial part of Astragalus mongholicus Bunge[J]. Chem Pharm Bul,1992,40(8):2230-2232.