大肠杆菌血清抗体与硫酸庆大霉素偶联构建抗体靶向药物的研究
|
摘要
近年来,治疗肿瘤的抗体药物研发取得了突破性的进展,抗体药物成为生物技术药物领域的热点之一。肿瘤抗体靶向药物主要是应用抗原抗体反应的原理构建肿瘤抗体药物,将治疗肿瘤的药物与抗体结合,构建“生物导弹”,对肿瘤实施生物疗法。而应用抗原抗体反应的原理构建抗菌药物,将抗菌药物与抗体结合,构建“生物导弹”,对病原菌实施“精确打击”的研究,却至今没有报道。为构建针对大肠杆菌的靶向性硫酸庆大霉素,本文进行了如下研究。
1.大肠杆菌血清抗体与聚乙二醇和硫酸庆大霉素复合物的制备
为了探讨大肠杆菌抗血清能否与硫酸庆大霉素结合,以及结合后是否会影响硫酸庆大霉素的药效和大肠杆菌血清抗体的生物学活性。本实验首先采用猪大肠杆菌灭活油乳佐剂苗,多次多部位背部皮下注射健康家兔,心脏采血,采集超免血清,用ELISA方法对抗猪大肠杆菌血清进行检测,其超免血清的抗体效价高达1:2048。然后对二者的结合条件进行摸索,以不同浓度的硫酸庆大霉素、免疫血清、PEG,建立方阵试验。收集各试验组血清抗体与硫酸庆大霉素结合产生的沉淀物进行聚丙烯酰胺电泳、革兰氏染色、电镜观察。结果发现,药物与血清抗体结合良好,以硫酸庆大霉素800(mL):免疫血清4(μg):PEG18(g)的结合量最大。因此,血清抗体与硫酸庆大霉素的结合具备有效性和可行性。
2.大肠杆菌血清抗体与PEG6000和硫酸庆大霉素复合物结构研究
为了探讨了结合物的结构,本实验对结合物进行了红外扫描和紫外吸收光谱测定,结果显示:结合物是以PEG6000为中间媒介。血清抗体与,硫酸庆大霉素先分别与PEG6000结合,从而形成血清抗体-PEG6000-硫酸庆大霉素复合物。结合的方式是氢键。PEG6000主要与血清抗体的酰胺键结合。
3.大肠杆菌血清抗体与PEG6000和硫酸庆大霉素复合物的靶向性研究
为了探讨结合物的靶向性,本实验对结合物进行间接ELISA、荧光染色和免疫电镜检测。间接ELISA结果表明:二者的结合对抗体的生物学活性和免疫学功能不会产生抑制作用;荧光抗体染色结果表明:硫酸庆大霉素、大肠杆菌血清抗体和PEG6000结合物具有大肠杆菌特异靶向性。硫酸庆大霉素、大肠杆菌血清抗体和PEG6000结合物与大肠杆菌菌液混合4min后,硫酸庆大霉素、大肠杆菌血清抗体和PEG6000结合物能与大肠杆菌结合。硫酸庆大霉素、大肠杆菌血清抗体和PEG6000结合物与大肠杆菌菌液混合7min后,硫酸庆大霉素、大肠杆菌血清抗体和PEG6000结合物能进入大肠杆菌菌体内;由胶体金免疫电镜结果中,在大肠杆菌的菌体内外均可见到胶体金颗粒,证明结合了药物的抗体的仍能高效与大肠杆菌作用,有较强的靶向作用。
4.大肠杆菌血清抗体与PEG6000和硫酸庆大霉素复合物药动学检测及安全性评价
本实验还探讨了结合物在动物体内的药动学并对其进行了安全性检测。结果表明:血清抗体-PEG6000-硫酸庆大霉素结合物在家兔体内的生物半衰期t_(1/2)为4.83 h,药-时曲线下面积AUG为191.41 h.μg/mL,硫酸庆大霉素结合物在家兔体内的生物半衰期为t_(1/2)1.03 h,AUC为63.42 h.μg/ml,这表明血清抗体-PEG6000-硫酸庆大霉素结合物药代动力学及组织分布发生了明显变化,药物代谢速度大大减慢,有效血药浓度时间延长,与脂质体缓释剂的特点相似。由于血清抗体-PEG6000-硫酸庆大霉素结合物毒性很低,对小鼠的最大耐受量大于750mg/kg.bw,此量相当于临床用量的150倍。结果表血清抗体-PEG6000-硫酸庆大霉素结合物的毒性显著低于硫酸庆大霉素,表明血清抗体-PEG6000有降低药物毒性的作用。血清抗体-PEG6000-硫酸庆大霉素结合物对小鼠重要脏器的生长发育无明显影响。
5.大肠杆菌血清抗体与PEG6000和硫酸庆大霉素复合物的药效学研究
为了研究结合物的药效学,本实验应用结合物进行了体外抗菌试验、大肠杆菌病动物模型治疗试验。
体外抑菌试验表明:靶向庆大霉素对大肠杆菌C_(44103)的最低抑菌浓度约为硫酸庆大霉素的1/10~5。靶向硫酸庆大霉素对大肠杆菌C_(44103)的最低杀菌浓度约为硫酸庆大霉素的1/10~4。体外抑菌实验说明将硫酸庆大霉素与大肠杆菌血清抗体和PEG6000结合后可增强抗菌效果,减少使用剂量,提高疗效。
动物模型治疗试验表明:用高浓度靶向庆大霉素注射液每次肌注1.0mL/只,一次用药后的有效率、治愈率分别为100%和90%;用中等浓度靶向庆大霉素注射液每次肌注1.0mL/只,一次用药后的有效率、治愈率分别为100%和75%;用低浓度靶向庆大霉素注射液每次肌注1mL/只,一次用药后的有效率为100%,治愈率为65%。而硫酸庆大霉素对照组用硫酸庆大霉素注射液每次肌注1.0mL/只,后发病小白鼠治疗无效或全部死亡;血清对照组也治疗无效。靶向庆大霉素注射液对小白鼠大肠杆菌病疗效显著。
复合物治疗仔猪黄痢临床治疗试验表明:靶向庆大霉素注射液能迅速杀死仔猪体内的致病菌,减轻临床症状,提高仔猪成活率。与硫酸庆大霉素相比较,用高剂量靶向庆大霉素注射液每次肌注2.0mg/Kg,一次用药后发病仔猪的有效率、治愈率均为100%;用中剂量靶向庆大霉素注射液每次肌注0.2mg/Kg,一次用药后发病仔猪的有效率为100%,治愈率为96%;用低剂量靶向庆大霉素注射液每次肌注0.02mg/Kg,两次用药后发病仔猪的有效率为96%,治愈率为92%;而用硫酸庆大霉素注射液每次肌注2.00mg/Kg,每天2次,连用3d后发病仔猪的有效率为84%,治愈率为76%;且用高剂量靶向庆大霉素注射液、中剂量靶向庆大霉素注射液、低剂量靶向庆大霉素注射液治疗的仔猪体内致病菌减少速度均要比硫酸庆大霉素注射液快得多。抗血清与庆大霉素结合物注射液能有效杀灭致病性大肠杆菌,对仔猪黄痢病有很好的临床疗效。
复合物治疗鸡大肠杆菌病临床治疗试验表明:抗血清与庆大霉素复合物各剂量组给药1d,病鸡病症均明显减轻,高剂量组第2d鸡停止死亡,鸡的食欲和精神基本恢复;中剂量组治疗效果与高剂量组的治疗效果基本一致;低剂量组第2d鸡停止死亡,腹泻症状基本消失,鸡的食欲与精神状态第3d也基本恢复。硫酸庆大霉素组给药第2d症状开始减轻,到第4d腹泻症状基本消失,食欲和精神状态到第5d恢复。从上表数据可以看出:抗血清与庆大霉素结合物注射液高、中、低三种剂量对鸡大肠杆菌病疗效显著,有效率和治愈率分别为100%,100%,92%;100%,100%,90%。而硫酸庆大霉素组的有效率和治愈率分别为80%;70%。就症状变化情况看,抗血清与庆大霉素结合物各剂量组给药后腹泻症状改善速度和鸡的精神状态恢复速度较快,和硫酸庆大霉素组相比能更快地改善鸡腹泻症状和恢复鸡的精神状态。
In recent years,The antibody treatment of cancer drug research and development has made breakthrough progress,it become a hot spot of The field of biotechnology drugs.Application of the principle of aritigen-antibody reaction Construction tumor antibody drugs,antibody and medicine combine a "biological missile",As Biological therapies of the therapy on tumor.Application of the principle of antigen-antibody reaction preparation of antibacterial drugs,antibody and antibiotics combine a "biological missile",As Biological therapies of the therapy on Bacterial Diseases,has not reported.This study coupled Serum antibody against E.coli with gentamicin sulfate
1.E.coli serum antibody and polyethylene glycol and gentamicin sulfate complex Preparation
To investigate whether the E.coli antisera with gentamicin sulfate,and the combination will affect the efficacy of gentamycin sulfate and E.coli serum antibody biological activity.The first experiments using pig oil emulsion inactivated E.coli adjuvant miller,many multi-site back healthy rabbits subcutaneously,heart blood collected serum - free,using ELISA method confrontation pig serum to detect E.coli, the serum - free up to 1:2048 antibody titer.Then the combination of the two conditions for exploration to different concentrations of gentamicin sulfate,sera,PEG, a test matrix.The collection of serum antibody test gentamicin sulfate and the combination of precipitation a polyacrylamide electrophoresis,Gram staining electron microscopy.The results found that combining drugs and serum antibody good to gentamicin sulfate 800(mg):sera4(μg):PEG18(g)the combination of the largest. Thus,the serum antibody and the combination with gentamicin sulfate effectiveness and feasibility.
2.E.coli serum antibody and PEG6000 and gentamicin sulfate complex structure
To explore the combination0f structure,the experimental combination of a pair of infrared scanning and UV absorption spectrometry showed that:a combination of PEG6000 as intermediary.Serum antibody and,first of gentamicin sulfate combined with PEG6000,thus forming serum antibody - PEG6000 - gentamicin sulfate complex. The combination of hydrogen bonds.PEG6000 mainly with the amide bond of serum antibody binding.
3.E.coli serum antibody and PEG6000 and gentamiein sulfate complexes targeted research
To explore the combination of targeting the right~ combination of experimental indirect ELISA,fluorescence staining of electron microscopy and immunohistochemistry.ELISA results show that:the combination of both the biological activity and antibody immune function does not produce inhibition; Fluorescent antibody staining results show that:gentamicin sulfate,E.coli and PEG6000 serum antibody binding of a specific targeting of E.coli.Gentamicin sulfate, E.coli and serum antibody binding of PEG6000 mixed with the E.coli bacterium after 4 min,gentamycin sulfate,E.coli and serum antibody binding of PEG6000 combined with E.coti.Gentamicin sulfate,E.coli and serum antibody binding of PEG6000 mixed with the E.coli bacterium after 7 min,gentamycin sulfate,E.coli and serum antibody binding PEG6000 of E.coli bacteria can enter the body;Colloidal gold by immuno-electron microscopy,in the E.coli bacteria can be seen inside and outside of colloidal gold particles that combining the antibody drugs still efficient and E.coli, targeting a strong role.
4.E.coil serum antibody and PEG6000 and gentamicin sulfate complex pharmacokinetics testing and evaluation of its safety
This study also explored the combination of in vivo pharmacokinetics and safety of its detection.The results show that:Serum antibody - Gentamicin Sulfate - PEG6000 in rabbits combination of the biological half-life t1/2 to 4.83 h,medicine - AUC,area under the curve for 191.41 h.μg / mL,according to the literature reported that the sulfate - adriamycin combination of rabbits in the biological half-life of t1/21.03 h, AUC 63.42 h.μg / ml,indicating that serum antibodies - PEG6000 combination of gentamicin sulfate- pharmacokinetics and tissue distribution in a significant change in the rate of drug metabolism the slower,effective plasma concentration time,and the release of liposome similar characteristics.The serum antibody - Gentamicin Sulfate -PEG6000 with low toxicity of the mice maximal tolerance than 750 mg / kg.bw,the equivalent of 150 time the amount.Results Table serum antibody - Gentamicin Sulfate PEG6000-toxic combination of gentamicin sulfate was significantly lower than that of serum antibody - PEG6000 reduction in the role of drug toxicity.Serum antibody -Gentamicin Sulfate PEG6000-right combination of the important organs of mice had no obvious impact on the growth and development
5.E.coli serum antibody and PEG6000 gentamicin sulfate complex and Pharmacodynamic Study
In order to study the combination of pharmacodynamics,the experimental application of a combination of in vitro antibacterial tests,E.coli disease animal model for experiments.
In vitro experiments showed that:Targeting gentamicin right E.coli C44103 the minimum inhibitory concentration of Gentamicin Sulfate about 1 / 10~5.Gentamicin Sulfate targeting of Escherichiacoli_(C44103)minimum bactericidal gentamicin sulfate concentration is about 1 / 10~3.In vitro experiments that will gentamicin sulfate and E. coli serum antibody and PEG6000 with antibacterial effect can be enhanced to reduce the dose to enhance efficacy.
Animal experiments showed that the treatment model:high concentration of targeting each intramuscular gentamicin injection mL / only one drug after an efficient, the cure rates were 100%and 90%;Medium concentrations targeted by gentamicin intramuscular injection every lmL / only one with after the drug is efficient,the cure rates were 100%and 75%;Targeting low concentration intramuscular gentamicin injection each lmL / only once after treatment with 100%efficiency,the cure rate was 65%.The control group was treated with gentamicin sulfate gentamicin sulfate intramuscular injection per mL/only,after the onset of void or mice all died;Serum is ineffective treatment for the control group.Targeting gentamicin injection of E.coli in mice significantly.
Clinical tests showed that:Targeting gentamicin injection can quickly kill the bacteria in pigs,reduce symptoms and improve the survival rate of piglets.Compared with gentamicin sulfate,gentamicin targeting high-dose intramuscular injection every 2.0mg/Kg,one after treatment onset piglets efficient,the cure rate was 100%;Targeting use of gentamicin dose intramuscular injection every 0.2mg/Kg,a drug after the piglets incidence rate was 100%,the cure rate is 96%;Targeting low-dose intramuscular gentamicin injection each 0.02mg/Kg two piglets after treatment morbidity rate was 96%,the cure rate was 92%;The gentamicin sulfate injection each injection 2.00mg/Kg,two times a day,3 days after the onset linked piglets efficiency of 84%,the cure rate is 76%;by targeting high-dose and gentamicin injection,the dose of gentamicin injection targeting,targeting low dose of gentamicin injection in the treatment of disease in piglets reduce speed than all of Gentamicin Sulfate Injection faster.Antisera combined with gentamicin injection can effectively kill the pathogenic E.coli,the piglets yellowscour of good clinical efficacy.
Complex treatment of E.coli in chicken clinical trial showed that:anti-serum and the complex gentamicin dose administration ld,diseased disease were significantly reduced,high-dose group,2d stop dead chickens,chicken and the spirit of appetite returned;Effect of dose and high dose treatment group basic treatment consistent; low-dose group,2d stop chicken deaths,diarrhea disappeared,chicken appetite and mental state of the 3d also recovered.Gentamicin sulfate group of 2d administration began to reduce symptoms,the symptoms disappeared 4d diarrhea,loss of appetite and mental state to restore the 5d.Data from the table can be seen:anti-serum and gentamicin injection with high,medium and low doses of three pairs of chicken E.coli disease significantly,efficient and cure rates were 100%,100%,92%;100%,100%, 90%.Gentamicin sulfate group and the efficiency and the cure rate was 80%;70%.The changes in symptoms,antisera and gentamicin combination of all dose groups after administration diarrhea improve speed and the mental state of the resumption of chicken faster,and gentamicin sulfhte compared to faster improvement in the chicken diarrhea and restore chicken mental state.
1.大肠杆菌血清抗体与聚乙二醇和硫酸庆大霉素复合物的制备
为了探讨大肠杆菌抗血清能否与硫酸庆大霉素结合,以及结合后是否会影响硫酸庆大霉素的药效和大肠杆菌血清抗体的生物学活性。本实验首先采用猪大肠杆菌灭活油乳佐剂苗,多次多部位背部皮下注射健康家兔,心脏采血,采集超免血清,用ELISA方法对抗猪大肠杆菌血清进行检测,其超免血清的抗体效价高达1:2048。然后对二者的结合条件进行摸索,以不同浓度的硫酸庆大霉素、免疫血清、PEG,建立方阵试验。收集各试验组血清抗体与硫酸庆大霉素结合产生的沉淀物进行聚丙烯酰胺电泳、革兰氏染色、电镜观察。结果发现,药物与血清抗体结合良好,以硫酸庆大霉素800(mL):免疫血清4(μg):PEG18(g)的结合量最大。因此,血清抗体与硫酸庆大霉素的结合具备有效性和可行性。
2.大肠杆菌血清抗体与PEG6000和硫酸庆大霉素复合物结构研究
为了探讨了结合物的结构,本实验对结合物进行了红外扫描和紫外吸收光谱测定,结果显示:结合物是以PEG6000为中间媒介。血清抗体与,硫酸庆大霉素先分别与PEG6000结合,从而形成血清抗体-PEG6000-硫酸庆大霉素复合物。结合的方式是氢键。PEG6000主要与血清抗体的酰胺键结合。
3.大肠杆菌血清抗体与PEG6000和硫酸庆大霉素复合物的靶向性研究
为了探讨结合物的靶向性,本实验对结合物进行间接ELISA、荧光染色和免疫电镜检测。间接ELISA结果表明:二者的结合对抗体的生物学活性和免疫学功能不会产生抑制作用;荧光抗体染色结果表明:硫酸庆大霉素、大肠杆菌血清抗体和PEG6000结合物具有大肠杆菌特异靶向性。硫酸庆大霉素、大肠杆菌血清抗体和PEG6000结合物与大肠杆菌菌液混合4min后,硫酸庆大霉素、大肠杆菌血清抗体和PEG6000结合物能与大肠杆菌结合。硫酸庆大霉素、大肠杆菌血清抗体和PEG6000结合物与大肠杆菌菌液混合7min后,硫酸庆大霉素、大肠杆菌血清抗体和PEG6000结合物能进入大肠杆菌菌体内;由胶体金免疫电镜结果中,在大肠杆菌的菌体内外均可见到胶体金颗粒,证明结合了药物的抗体的仍能高效与大肠杆菌作用,有较强的靶向作用。
4.大肠杆菌血清抗体与PEG6000和硫酸庆大霉素复合物药动学检测及安全性评价
本实验还探讨了结合物在动物体内的药动学并对其进行了安全性检测。结果表明:血清抗体-PEG6000-硫酸庆大霉素结合物在家兔体内的生物半衰期t_(1/2)为4.83 h,药-时曲线下面积AUG为191.41 h.μg/mL,硫酸庆大霉素结合物在家兔体内的生物半衰期为t_(1/2)1.03 h,AUC为63.42 h.μg/ml,这表明血清抗体-PEG6000-硫酸庆大霉素结合物药代动力学及组织分布发生了明显变化,药物代谢速度大大减慢,有效血药浓度时间延长,与脂质体缓释剂的特点相似。由于血清抗体-PEG6000-硫酸庆大霉素结合物毒性很低,对小鼠的最大耐受量大于750mg/kg.bw,此量相当于临床用量的150倍。结果表血清抗体-PEG6000-硫酸庆大霉素结合物的毒性显著低于硫酸庆大霉素,表明血清抗体-PEG6000有降低药物毒性的作用。血清抗体-PEG6000-硫酸庆大霉素结合物对小鼠重要脏器的生长发育无明显影响。
5.大肠杆菌血清抗体与PEG6000和硫酸庆大霉素复合物的药效学研究
为了研究结合物的药效学,本实验应用结合物进行了体外抗菌试验、大肠杆菌病动物模型治疗试验。
体外抑菌试验表明:靶向庆大霉素对大肠杆菌C_(44103)的最低抑菌浓度约为硫酸庆大霉素的1/10~5。靶向硫酸庆大霉素对大肠杆菌C_(44103)的最低杀菌浓度约为硫酸庆大霉素的1/10~4。体外抑菌实验说明将硫酸庆大霉素与大肠杆菌血清抗体和PEG6000结合后可增强抗菌效果,减少使用剂量,提高疗效。
动物模型治疗试验表明:用高浓度靶向庆大霉素注射液每次肌注1.0mL/只,一次用药后的有效率、治愈率分别为100%和90%;用中等浓度靶向庆大霉素注射液每次肌注1.0mL/只,一次用药后的有效率、治愈率分别为100%和75%;用低浓度靶向庆大霉素注射液每次肌注1mL/只,一次用药后的有效率为100%,治愈率为65%。而硫酸庆大霉素对照组用硫酸庆大霉素注射液每次肌注1.0mL/只,后发病小白鼠治疗无效或全部死亡;血清对照组也治疗无效。靶向庆大霉素注射液对小白鼠大肠杆菌病疗效显著。
复合物治疗仔猪黄痢临床治疗试验表明:靶向庆大霉素注射液能迅速杀死仔猪体内的致病菌,减轻临床症状,提高仔猪成活率。与硫酸庆大霉素相比较,用高剂量靶向庆大霉素注射液每次肌注2.0mg/Kg,一次用药后发病仔猪的有效率、治愈率均为100%;用中剂量靶向庆大霉素注射液每次肌注0.2mg/Kg,一次用药后发病仔猪的有效率为100%,治愈率为96%;用低剂量靶向庆大霉素注射液每次肌注0.02mg/Kg,两次用药后发病仔猪的有效率为96%,治愈率为92%;而用硫酸庆大霉素注射液每次肌注2.00mg/Kg,每天2次,连用3d后发病仔猪的有效率为84%,治愈率为76%;且用高剂量靶向庆大霉素注射液、中剂量靶向庆大霉素注射液、低剂量靶向庆大霉素注射液治疗的仔猪体内致病菌减少速度均要比硫酸庆大霉素注射液快得多。抗血清与庆大霉素结合物注射液能有效杀灭致病性大肠杆菌,对仔猪黄痢病有很好的临床疗效。
复合物治疗鸡大肠杆菌病临床治疗试验表明:抗血清与庆大霉素复合物各剂量组给药1d,病鸡病症均明显减轻,高剂量组第2d鸡停止死亡,鸡的食欲和精神基本恢复;中剂量组治疗效果与高剂量组的治疗效果基本一致;低剂量组第2d鸡停止死亡,腹泻症状基本消失,鸡的食欲与精神状态第3d也基本恢复。硫酸庆大霉素组给药第2d症状开始减轻,到第4d腹泻症状基本消失,食欲和精神状态到第5d恢复。从上表数据可以看出:抗血清与庆大霉素结合物注射液高、中、低三种剂量对鸡大肠杆菌病疗效显著,有效率和治愈率分别为100%,100%,92%;100%,100%,90%。而硫酸庆大霉素组的有效率和治愈率分别为80%;70%。就症状变化情况看,抗血清与庆大霉素结合物各剂量组给药后腹泻症状改善速度和鸡的精神状态恢复速度较快,和硫酸庆大霉素组相比能更快地改善鸡腹泻症状和恢复鸡的精神状态。
In recent years,The antibody treatment of cancer drug research and development has made breakthrough progress,it become a hot spot of The field of biotechnology drugs.Application of the principle of aritigen-antibody reaction Construction tumor antibody drugs,antibody and medicine combine a "biological missile",As Biological therapies of the therapy on tumor.Application of the principle of antigen-antibody reaction preparation of antibacterial drugs,antibody and antibiotics combine a "biological missile",As Biological therapies of the therapy on Bacterial Diseases,has not reported.This study coupled Serum antibody against E.coli with gentamicin sulfate
1.E.coli serum antibody and polyethylene glycol and gentamicin sulfate complex Preparation
To investigate whether the E.coli antisera with gentamicin sulfate,and the combination will affect the efficacy of gentamycin sulfate and E.coli serum antibody biological activity.The first experiments using pig oil emulsion inactivated E.coli adjuvant miller,many multi-site back healthy rabbits subcutaneously,heart blood collected serum - free,using ELISA method confrontation pig serum to detect E.coli, the serum - free up to 1:2048 antibody titer.Then the combination of the two conditions for exploration to different concentrations of gentamicin sulfate,sera,PEG, a test matrix.The collection of serum antibody test gentamicin sulfate and the combination of precipitation a polyacrylamide electrophoresis,Gram staining electron microscopy.The results found that combining drugs and serum antibody good to gentamicin sulfate 800(mg):sera4(μg):PEG18(g)the combination of the largest. Thus,the serum antibody and the combination with gentamicin sulfate effectiveness and feasibility.
2.E.coli serum antibody and PEG6000 and gentamicin sulfate complex structure
To explore the combination0f structure,the experimental combination of a pair of infrared scanning and UV absorption spectrometry showed that:a combination of PEG6000 as intermediary.Serum antibody and,first of gentamicin sulfate combined with PEG6000,thus forming serum antibody - PEG6000 - gentamicin sulfate complex. The combination of hydrogen bonds.PEG6000 mainly with the amide bond of serum antibody binding.
3.E.coli serum antibody and PEG6000 and gentamiein sulfate complexes targeted research
To explore the combination of targeting the right~ combination of experimental indirect ELISA,fluorescence staining of electron microscopy and immunohistochemistry.ELISA results show that:the combination of both the biological activity and antibody immune function does not produce inhibition; Fluorescent antibody staining results show that:gentamicin sulfate,E.coli and PEG6000 serum antibody binding of a specific targeting of E.coli.Gentamicin sulfate, E.coli and serum antibody binding of PEG6000 mixed with the E.coli bacterium after 4 min,gentamycin sulfate,E.coli and serum antibody binding of PEG6000 combined with E.coti.Gentamicin sulfate,E.coli and serum antibody binding of PEG6000 mixed with the E.coli bacterium after 7 min,gentamycin sulfate,E.coli and serum antibody binding PEG6000 of E.coli bacteria can enter the body;Colloidal gold by immuno-electron microscopy,in the E.coli bacteria can be seen inside and outside of colloidal gold particles that combining the antibody drugs still efficient and E.coli, targeting a strong role.
4.E.coil serum antibody and PEG6000 and gentamicin sulfate complex pharmacokinetics testing and evaluation of its safety
This study also explored the combination of in vivo pharmacokinetics and safety of its detection.The results show that:Serum antibody - Gentamicin Sulfate - PEG6000 in rabbits combination of the biological half-life t1/2 to 4.83 h,medicine - AUC,area under the curve for 191.41 h.μg / mL,according to the literature reported that the sulfate - adriamycin combination of rabbits in the biological half-life of t1/21.03 h, AUC 63.42 h.μg / ml,indicating that serum antibodies - PEG6000 combination of gentamicin sulfate- pharmacokinetics and tissue distribution in a significant change in the rate of drug metabolism the slower,effective plasma concentration time,and the release of liposome similar characteristics.The serum antibody - Gentamicin Sulfate -PEG6000 with low toxicity of the mice maximal tolerance than 750 mg / kg.bw,the equivalent of 150 time the amount.Results Table serum antibody - Gentamicin Sulfate PEG6000-toxic combination of gentamicin sulfate was significantly lower than that of serum antibody - PEG6000 reduction in the role of drug toxicity.Serum antibody -Gentamicin Sulfate PEG6000-right combination of the important organs of mice had no obvious impact on the growth and development
5.E.coli serum antibody and PEG6000 gentamicin sulfate complex and Pharmacodynamic Study
In order to study the combination of pharmacodynamics,the experimental application of a combination of in vitro antibacterial tests,E.coli disease animal model for experiments.
In vitro experiments showed that:Targeting gentamicin right E.coli C44103 the minimum inhibitory concentration of Gentamicin Sulfate about 1 / 10~5.Gentamicin Sulfate targeting of Escherichiacoli_(C44103)minimum bactericidal gentamicin sulfate concentration is about 1 / 10~3.In vitro experiments that will gentamicin sulfate and E. coli serum antibody and PEG6000 with antibacterial effect can be enhanced to reduce the dose to enhance efficacy.
Animal experiments showed that the treatment model:high concentration of targeting each intramuscular gentamicin injection mL / only one drug after an efficient, the cure rates were 100%and 90%;Medium concentrations targeted by gentamicin intramuscular injection every lmL / only one with after the drug is efficient,the cure rates were 100%and 75%;Targeting low concentration intramuscular gentamicin injection each lmL / only once after treatment with 100%efficiency,the cure rate was 65%.The control group was treated with gentamicin sulfate gentamicin sulfate intramuscular injection per mL/only,after the onset of void or mice all died;Serum is ineffective treatment for the control group.Targeting gentamicin injection of E.coli in mice significantly.
Clinical tests showed that:Targeting gentamicin injection can quickly kill the bacteria in pigs,reduce symptoms and improve the survival rate of piglets.Compared with gentamicin sulfate,gentamicin targeting high-dose intramuscular injection every 2.0mg/Kg,one after treatment onset piglets efficient,the cure rate was 100%;Targeting use of gentamicin dose intramuscular injection every 0.2mg/Kg,a drug after the piglets incidence rate was 100%,the cure rate is 96%;Targeting low-dose intramuscular gentamicin injection each 0.02mg/Kg two piglets after treatment morbidity rate was 96%,the cure rate was 92%;The gentamicin sulfate injection each injection 2.00mg/Kg,two times a day,3 days after the onset linked piglets efficiency of 84%,the cure rate is 76%;by targeting high-dose and gentamicin injection,the dose of gentamicin injection targeting,targeting low dose of gentamicin injection in the treatment of disease in piglets reduce speed than all of Gentamicin Sulfate Injection faster.Antisera combined with gentamicin injection can effectively kill the pathogenic E.coli,the piglets yellowscour of good clinical efficacy.
Complex treatment of E.coli in chicken clinical trial showed that:anti-serum and the complex gentamicin dose administration ld,diseased disease were significantly reduced,high-dose group,2d stop dead chickens,chicken and the spirit of appetite returned;Effect of dose and high dose treatment group basic treatment consistent; low-dose group,2d stop chicken deaths,diarrhea disappeared,chicken appetite and mental state of the 3d also recovered.Gentamicin sulfate group of 2d administration began to reduce symptoms,the symptoms disappeared 4d diarrhea,loss of appetite and mental state to restore the 5d.Data from the table can be seen:anti-serum and gentamicin injection with high,medium and low doses of three pairs of chicken E.coli disease significantly,efficient and cure rates were 100%,100%,92%;100%,100%, 90%.Gentamicin sulfate group and the efficiency and the cure rate was 80%;70%.The changes in symptoms,antisera and gentamicin combination of all dose groups after administration diarrhea improve speed and the mental state of the resumption of chicken faster,and gentamicin sulfhte compared to faster improvement in the chicken diarrhea and restore chicken mental state.
引文
[1]Bangham AD,Standish MN,Watkins JC.The action of steroid and streptolysin Son the permeability of phospolipic structures to cations[J].J Mol Biol,1965,13:138.
[2]Rahman YE,Cerny EA,Tollaksen SL,et al.Liposome-encapsulated actinoycin D:potential in cancer chemotherapy.Proc Soc Exp BiolMed.1974,146(4):1173-1176
[3]Fouda HG.Determination of diminazene in plasma by high-perforarnnce liquid chromatography[J].Journal of Chromatographic Science,1977,15:537-538
[4]L.Meyer Jones,Nicholas H.Booth,Leslie E.McDonald主编.兽医药理学与治疗学[M],冯淇辉,中葆和,戎耀方,等泽.上海科学文献出版社,1982:727
[5]Tsutumi F.Sakakibara M,Oshima N.Structure and dynamicproperties of solution SBR coupled with tin compounds[J].Rubber Chem istry and Technology.1990,69(1):8
[6]Alzheimer disease:evidence for chromosome 19 linkage[J].Ame J Hum Genet,1991,48(5):1034
[7]Hall RM,brookesDE,stokesHW.Site-specific Insertion of Genes into Integrons:Role of the 59-base Element and Determina-tion of the Recombination Cross-over point[J].Mol Microbiol,1991,5:1941-1953
[8]Murray BE.New aspects of antimicrobial resistance and resulting the therapeutic dilem mas[J]InfectDis,1991,163:1185
[9]Fattal E,Rojas J,Youssef M,et al.Liposome-entrapped ampicilin in the treatment of experimental murine listeriosis and salmonellosis[J].Antimicrob Agents Chemother,1991,35(4):770
[10]Tomioka H,Saito H,Sato K,et al.Therapeutic efficacy of liposome-encapsulated kanamycinagainst mycobaterium intracellulare infection induced in mice[J].Am Rev Respir Dis,1991,144(3):575
[11]Yeh MY.R(,filer SR.Doxorubicin:monoclonal antibody conjugate for therapy of human cervical carcinomal[J].lnt J Cancer.1992,51(2),274-282
[12]Woodle MC,Lasic DD.Sterically stabilized liposomes[J].Biochim Biophys Acta,1992,1113(2):171-199
[13]Ana cabanes,Margarita arboix,Jose M,et al.Pharmacoki-netics of enrofloxacin after intravenous and intramuscular in-jection in rabbits[J].Am J Vet Res,1992,53(11):2090-2093
[14] Mehta RT, Kevhani A, Mequeen T.I, et al. In vitro activities of free and liposomal drugs against Mvcobacterium avium-M, intracellulare complex and M. tuberculonis[J]. Antimicrob Agents Chemother,1993,37(12):2584
[15]Pastenack R F, BustamanteC, Collings P J. Am. Chemv.Soci, 1993, 115: 5393-5399
[16] Forssen EA, Ross ME. Daunoxome treatment of solid turners :preclinical and clinical investigations[J]. J Liposome Res , 1994 ,4 (1): 481—512
[17] Yuan F, Dellian M, FukumuraD, et al. Vascular permeability in human tuman tumor xenograft molecular size dependence and cutoff size [J].Cancer Res, 1995. 55: 3752—3756.
[18]SakaeU, KazuoM, Osamu I, et al.Enhanced tumor targeting and improved antitumor activity of doxorubi by cin long-circulating liposomes containing amphipathic poly(ethylene glycol)[J]. International Journal of Pharmaceutics, 1995, 126: 41 —48
[19] Zhu Z, Kralovec J Inhibition of epstein—Barr—virus—transformed human chornic lymphocytic leukaemic B cells with monoclonal antibody adriamycin (doxorubicin) conjugates [J]. Cancer Immunol Immunother, 1995, 40(4): 257—267
[20] Hsiech H L, Quirk R P. Anionic polymerization: principles and practical applications[J]. New York: Marcel Dekker Inc. 1996. 447
[21] Mendelsohn L, Epidermal growth factor receptor inhibition by a monoclonal antibody as antibody therapy[J] Clin( ncer Res, 1997, 3(12): 2703—2707
[22] Pardue R L, White C A. Pharmacokinetic evaluation of liposomal encapsulated ampicillin in male and female rats[J]. Biopharm Drug Dispos 1997,18(4):279-292.
[23] Vladimir A,Slepushkin, Sengio S,et al.Sterically stabilized pH-sensitive liposomes[J]. Journ Biol Chemi, 1997, 272: 2382-2388
[24] Pharmacokinetic Pardue R L, White C A. Pharmacokinetic evaluation of liposomal encapsulated ampicillin in male and female rats[J].Biopharm Drug Dispos, 1997,18(4):279—292.
[25] Wiechens B , Krausse R , Crammeer J B , et al. Clearance of liposome-incorporated ciprolloxacin after intravitreal injection in rabbit eyes[J]. Klin Monatsb Augenheilkd , 1998, 213(5):284-292.
[26] Omelyanenko V, Kopekuaa P Targetable HPMA copolymer-adriamycin conjugates, recognition, internalisation and subcellular fate[J]. J Controlled Release, 1998, 53(1—3): 25— 37.
[27] Reiter Y. Pastan 1 Recombinant Fv immunotoxins and Fv flag—ments as nox el agents for cancer therapy and diagnosis[J] J rends Biote～h, 1998, 16(12): 513—520
[28] Oussoren C, Velinova M, Scherphof G. et al. Lymphatic uptake and biodistribution of liposomes after subcutaneous injection. IV. Fate of liposomes in regional lymph nodes[J]. Biochim Biophys Acta, 1998, 1370: 259-272.
[20] Higuchi M, Fokin A, Masters TN, et al. Transport of colloidal particles in lymphatics and vasculature after subcutaneous injection[J]. J Appl Physiol, 1999. 86: 1381 — 1387
[30] Brocchini S, Duncma R Polymer drIJg conjugates: drug release from pendent linker: A Mathiowitz E(ED)Encyclopaedia of controlled drug delivery[M]. New York: Wiley, 1999: 786-816
[31] Takahashi H, Adacchi K. Experimental treatment of malignant gliomas with human monoclonal antibody drug conjugates[J]. AnticancerRes, 1999, 19(12): 4151—4156
[32] Ota Y, Fukasawa I Antitumor effect of monoc lonal antibody-car-boplatin conjugates in nude mice bearing ovarian cancer cells[J]. Int J Clin Oncol, 1999, 4(2): 236—240
[33] Ochs MM, Mc Cusker MP, Bains M, et al. Negative regulation of the Pseudomonas aeruginosa outer membrane porin OprD selective for imipenem and basic aminoacids [J]. Antimicrob Agents Chemother,1999, 43:1085—1090.
[34] Mckessar SJ, Berry AM,Bell JM, et al. Genetic Characterization of VanG,A novel vancomycin reisistance locus of Enterococcus fecalis [J] ,Antimicrob Agents Chemother,2000,44:3224— 3228.
[35] O samu I, Kazuo M, Hironobu Y, et al.Targeting chemotherapy to solid tumors with long-circulation thermosensitive liposomes and local hypertherthermia[J]. Jpn J Cancer Res, 2000, 91: 118-126
[36] Duncan R, Gac-Breton S. Keane R, et al Polymer-drug conjugates, PI)EI l and PELT: basic principles for design and transfer from the la}× )ratory to clinic[J]. J Controlled Release. 2001, 74(1—3)135 -146
[37] Garett MC. Iargeted drug conjugates: principles and progress [J].Adv Drug Deltev Rev, 2001, 53(2): 171-216
[38] Liebe V, Bruckmann M, Fischer KG, et al. Biologicalrelevance of anti recombinant hirudin antibodies-results from nvitro and in vivo studies[J]. Semin Thromb Hemost, 2002, 28(5): 483-490
[39] Phillips RK, Wallace MH, Lynch PM, et al. A randomized, double blind, placebo controlled study of celecoxib, a selective cyclooxygenase 2 inhibitor, on duodenal polyposis in familial adenomatous polyposis[J]. Gut, 2002, 50(6): 857-860
[40] Garberk, TeBexxar.FDA weeting offers long—awaitedchance at approvat[J].J Natl Cancer Inst. 2002, 94 (23) : 1738.
[41] Richardson PG, Barlogie B, Berenson MD, et al. A phase 2 study of bortezomib in relapsed, refractory myeloma[J]. N Engl J Med, 2003, 348(26): 2609-2617
[42] wusu-Asiedu A. et al. Response of early-weaned pigs to an enterotoxigenic Escherichia coli(K88) challenge when fed diets containing spray-dried porcine plasma or pea protein isolate plus egg yolk antibody [J] .J.Anim. Sci,2003,82: 1781-1789
[43] Nakata E,Hunter N,Mason K,et al.C225 antiepidermal growth factor receptor antibody enhances the efficacy of docetaxel chemoradiotherapy[J].Int J Radiat Oncol Biol Phys,2004,59(4): 1163 — 1173
[44] Martin Tulp, Lars Bohlin. Unconventional natural sources for future drug discovery [J]. Drug Discovery Today,2004,9( 10):9
[45] Mire-sluisa AR, Barrettb WC, Devanarayan V, etal. Recommendations for the design and optimization of immunoassaysused in the detection of host antibodies against biotechnol—ogy products[J]. J Immunol Methods, 2004, 289(1): 1 —16.
[46] Hurvitz H, Fehrenbacher L, NovotnyW, et al. Bevacizumab plus iinotecan, fluoruracil, and leucovorin for metasratic correctal[J].Nev Engl J Med.2004, 350 (23) : 2335
[47] Dittmann K,Mayer C,Fehrenbacher B,et al.Radiation-induced epidermal growth factor receptor nuclear import is linked to activation of DNA-dependent protein kinase.J Biol Chem,2005,280 (35):31182-31189
[48] Wadhwa M, Gaines-das R. Thorpe R, etal. Detection, measurement and characterization of unwanted antibodies inducedby therapeutic biologicals[J]. Dev Biol(Basel), 2005, 122: 155— 170.
[49] Shen B, Nonaka K, Nonaka K. Enediyne natural product: Biosynthesis and prospect towards engineering novel antitumor agents [J]. Bentham Science Publishers,2005,2 : 357-369
[50] Wan, Yinhua; Ghosh, Raja, Cui; Zhanfeng and Hale, Geoff. Fractionation of BSA and Monoclonal Antibody Alemtuzumab Using Carrier Phase Ultrafiltration [J]. Biotechnol Bioeng, 2005,90(3):303-315
[51] Feng Q, Yang J, Luo P,et al. LT(K63/R72),a new mutant of Escherichia coli heat-labile enterotoxin,exhibits characteristics more similar to LT(K63)than LT(R72)[J].Acta Biochim Biophys Sin(Shanghai),2005.37(2):126-132
[52]Pao W,Miller VA.Fpidermal growth factor receptor mutations,small-molecule kinase inhibitors,and non-smal1-cell-lung cancer:current knowledge and future directions.J Clin Oncol,2005,23(11):2556-2568.
[53]Burtness B.The role of cetuximab in the treatment of squamous cell cancer of the head and neck.Expert Opin Biol Ther,2005,5(8):1085-1093.
[54]Raizer JJ.HER1/EGFR tyrosine kinase inhibitors for the treatment of glioblastoma multiforme.J Neurooncol,2005,74(1):77-86.
[55]Harari PM,Huang S.Radiation combined with EGFR signal inhibitors:head and neck cancer focus.Semin Radiat Oncol,2006,16(1):38-44
[56]Tillmann HC,Kuhn B,Kranzlin B,et at.Efficacy andimmunogenicity of novel erythropoietic agents and conventionalrhEPO in rats with renal insufficiency[J].Kidney lnt,2006,69(1):60-67.
[57]Chang W Y.Carter R,Hardy T,etal.Inhibition of Escherichia coil heat-labile enterotoxin B subunit pentamer(EtxB5)assdmbly in vitro using monouonal antibodies[J].Biol chem.,2006,10:1074
[58]T.Luke Simmons and Kerry L.McPhail etc.Belamide A=Belamide A,a new antimitotic tetrapeptide from a Panamanian marine cyano- bacterium[J].Tetrahedron Letters,2006,47(20):3387-3390.
[59]Wilson GD,Bentzen SM,Harari PM.Biologic basis for combining drugs with radiation.Semin Radiat Oncol,2006,16(1):2-9
[60]施新猷.休克模型的复制[J].医学动物实验方法,1980,250-254
[61]胡远峰.大鼠糖尿病模型[J].中华内科杂志,1981,(9):535
[62]卢兴.儿种常用心律失常动物模型的复制[J].人类疾病的动物模型,1982,1(19):182-199
[63]重尔昌.大白鼠的实验性巨结肠[J].人类疾病的动物,1982,271(274):308-310
[64]吴荣庆.大白鼠胆管炎模型制作方法的介绍[J],武汉医学院学报,1983,(4):364
[65]夏家明.大鼠实验性急性肺水肿模型[J].上海实验动物科学,1985,5(4):240
[66]吴若铢.天花粉引起被动肺过敏的大鼠模型[J].中国药理学报,1985,6(1):68
[67]谢明智.实验性肥胖及糖尿病大鼠模型[J].药学学报,1985,20(11):801
[68]郭尧君.分光光度技术及其在生物化学中的应用[M].科学出版社,1987.230-231
[69]章文贡,陈田安,陈文定.铝酸酯偶联剂改性碳酸钙的性能与应用[J].中国塑料,1988,2(1):3
[70]Habamnh CM等.丁胺卡那霉素的特点平临床应用[J].国外医药抗生索分册,1989,10(5):365-368.
[71]何晓青.卫生防疫细菌检验[M].新华出版社1989 216-520
[72]容健材,廖锡麟.新编药理学[M].江苏科技出版社,1990:515
[73]傅永林.偶联剂住塑料复合材料中的应用[J].中国塑料.1991.5(3):20
[75]李战强.王永沛,李荣兴.等.双金属偶联剂FPM在电缆橡胶中的应用[J].无机盐工业.1992.120(5):40
[76]肖希龙,沈建忠,朱蓓蕾等.泰乐加和呋喃它酮对大肠杆菌与沙门氏菌的药效研究[J]畜牧兽医学报,1993,24(1):81-86
[77]王树槐,潘松年.盐酸环丙沙星注射液对仔猪大肠杆菌病的临床药效试验[J].中国兽药杂志,1994,28(3):34-35、
[78]徐美娟,聂春发等.临床常用氨基糖苷在烫伤兔体中的药动学改变[J].中国抗生素杂志,1994,19(1):42-45.
[79]张仰亨,张榕平.比色测定法测定硫酸庆大霉素及其注射液的含量[J].海峡药学,1994,6(3):27-28.
[80]何笑蓉。赵研.傅得兴.一阶导数光谱法测定硫酸庆大霉素制剂的含量[J].中国医院约学杂志。1995,15:(11):505-507
[81]李好枝,高光恩,田洁.荧光光谱法测定硫酸庆大霉素注射液的含量[J].西北药学杂志,1995,10(1):5-7
[82]李铜铃,庞其捷,贺于玲,等.肝靶向抗疟药半乳糖基似糖白蛋白-伯氨喹偶联物和磷酸伯氨喹的药代动力学[J].药学学报,1995,30(10):721-725
[83]李富荣,王琪,李守唐.吡喹酮和吡喹酮脂质体RP-HPLC法测定及药代动力学研究[J].中国药理学通报,1995,11(4):341-343
[84]陈士朝.轮胎的性能要求与合成橡胶_的发展[J].合成橡胶工业.1995.18(5):260
[85]张强,廖工铁,寿旦.用高效液相色谱法测定兔血清中硫酸庆大霉素的含量[J]中国医院药学杂志,1995,15(4):147-149.
[86]杨秀岑,伍莉萍。邓安平等.生物素标记增强化学发光法测定血清中庆大霉素含量[J].华西医大学报,1995,26(2):234-236
[87]张永恒.现代靶向制剂[M].黑龙江科学技术出版社,1995:1.
[88]丁鼎乐.靶向制剂国内外研究进展[J].中国医药情报,1996,2(5):301
[89]张强,廖工铁,魏大鹏等.用毫微球增加体外小鼠腹腔巨噬细胞与大鼠肝细胞对庆大霉素的摄取[J].药学学报,1996,31(5):375-380
[90]徐葛林,严家信.脂质体用作疫苗佐剂的研究进展[J].国外医学-预防、诊断、治疗用生物制品分册,1996,19(1):10.
[91]崔春英。席德荣.胃乐合剂中硫酸庆大霉素的含量测定.中国药事.1996。10(2):117.
[92]武德珍,白宗武.稀土偶联剂处理碳酸钙填充PVC复合材料及其性能的研究[J].现代塑料加工应用.1996,8(1):10.
[93]鲍爱华.世界溶聚丁苯橡胶的主要技术进展[J].石油化工动态,1996.4(3):18
[94]张开.高分子界面科学[M].中国石化出版社.1997.223
[95]陈士朝.溶聚丁苯橡胶的技术进展[J].合成橡胶工业.1997.20(1):6
[96]徐宝梁,李谨,马颖.有关杯碟法沁定壮观霉素灵敏度的影响因素[J].微生物学通报,1997,24(6):372-375
[97]王晶钰.陕西省鸡致病性大肠杆菌生物学特征和免疫防治研究[J].西北农林科技大学学报,1997,(2):97-101.
[98]廖工铁.靶向给药制剂[M].成都:四川科学技术出版社,1997:334-371.
[99]平其能.现代药剂学[M].中国医药科技出版社,1998:588-621.
[100]陆彬.药物新剂型与新技术[M].人民卫生出版社,1998:32,
[101]姜为东,黄引贤.广东省仔猪腹泻大肠杆菌病血清型鉴定[J].中国兽医学报,1998,18(1):63-65.
[102]张均田.现代药理实验方法[M].北京:北京医科大学中国协和医科大学联合出版社,1998:1412-1415
[103]刘继兰,李敬玺,袁有申等.“9381-杀灭剂”对仔猪大肠杆菌病的治疗试验[J].中国畜禽传染病,1998,20,(1):49-50
[104]顾觉奋,王鲁燕,张俊琪.阿米卡星脂质体在小鼠体内的组织分布[J].中国抗生素杂志,1998,23(4):294-297
[105]陆彬.药物新剂型与新技术[M].人民卫生出版社,1998:326-329
[106]梁亮,廖列文,崔英德.铝一锆有机金属络合物偶联剂的合成及应用[J].精细化工,1999.16(4):4
[107]高崧,刘秀梵,张如宽.我国部分地区禽病源性大肠杆菌的分离和鉴定[J].畜牧兽医学报,1999,30:164-171.
[108]王庆民,宋尚龙.用旋光法测定硫酸庆大霉素注射液的含量[J].药品检测,1999,8(7):45-46.
[109]舒泉.细菌DNA分析在流行病学中的应用[J].中华流行病学杂志,1999,(12):52-54
[110]张秀英.抗生素的耐药机制分类及概述[M].1999年全国兽医学会微生物学学会年会论文集,205-209
[111]何宁,王昌禄,顾晓波等.杯碟法检测乳酸菌素活性优化条件的研究[J].天津轻工业学院学报,1999,2:18-20
[112]甑永苏.单克隆抗体药物治疗肿瘤的研究现状及展望[J]中国医学科学院学报,2000,2(1):9-13
[113]江寅生,张皖华.庆大霉素滴鼻剂的研制及含量测定[J]中国药业,2000,9(10):53
[114]吴建良,沈特庞,陈晓萍.电荷转移络合--阶导数光谱法测定硫酸庆大霉素滴眼液的含最[J].西北药学杂志,2000.15(4):152-153
[115]朱立平,陈学清.免疫学常用实验技术[M].人民军医出版社,2000:57-61
[116]张永泉。杜战江,刘云等.d,JL硫酸庆大霉素颗粒含量测定方法研究[J].中国药事,2000,14(3):177-178.
[117]庞扶民。李静,张汉宁.用旋光法测定硫酸庆大霉素溶液的含量[J].中国煤炭工业医学杂志,2000.3(2):151
[118]姜为东,黄引贤.广东省仔猪腹泻大肠杆菌病血清型鉴定[J].中国兽医学报,2000,18(1):63-65
[119]刘又宁,王睿,邓宝军等.头孢呋辛钠治疗细菌性感染临床评价[J].中国临床药理字杂志,2000,16(4):251-254
[120]高德伟 王华琼 张进川 刘春雷.第四代头孢菌素头孢吡肟在老年肺炎治疗中的应用[J].中华医院感染学杂志,2000,10(3):221-222
[121]吕志堂 刘志恒 刘志恒.一种从土壤样品中选择性分离假诺卡氏菌的方法[J].微生物学报,2000,40(4):406-113
[122]鞠洪涛.大肠杆菌耐药基因定位及耐药质粒的消除[J].中国兽医学报,2000(6):2
[123]刘业兵,高松.禽病原性大肠杆菌Ⅰ型茵毛单克隆抗体的研制极其对分离株的检测[J].中国兽医学报,2000,20(2):148-151
[124]赵雨,徐金泽,赵大庆.二维拉曼相关光谱研究稀土离子对血红蛋白结构的影响[J].高等学校化学学报,2000,21(12):1913-1915
[125]俞英,程晓伟.牛血清白蛋白与酸性品红的结合反应[J].分析化学,2000,28(5):583- 586
[124]赵坤,王三虎.克痢注射液对仔猪大肠杆菌病治疗效果研究[J].中国医学杂志,2000,(3):8-9
[125]于波涛,张志荣,曾仁杰.约质体-一种新型给药系统[J].中国药学杂志,2000,35(11):721.
[126]彭应旭,廖工铁.骨髓靶向给药制剂的研究进展[J].中国药学杂志,2000,35(2):73
[127]张静,丁永红.木质素作为偶联剂在橡胶中的作用[J].特种橡胶制品,2001,22(6):22
[128]包晓玉,陈建国,陈欣,党元林.芦丁与DNA相互作用的电化学研究[J],分析试验室,2001,20(2):1-3
[129]凌连生,杨洗,何治柯,曾云鹗.小分子与DNA相互作用的光谱研究[J].分析科学学报,2001,17(1):11-15
[130]范盛先,袁宗辉,孔科等.庆大霉素在猪肉和鸡肉组织中残留的微生物学检测方法研究[J].华中农业大学学报,2001:557-560
[131]杨晓青,张强,吴霖.我国药剂学靶向制剂研究的基本思路[J].中国药学杂志,2001,36(12):797
[132]夏业才,姚文生,刘惠文.沙拉沙星注射液对人工诱发仔猪大肠杆菌病的临床疗效试验[J].中国兽药杂志,2001,35(1):66-67
[133]黄玲.胃宁冲剂中盐酸酱鲁卡因与硫酸庆大霉素的含量测定[J].中国医院药学杂志,2001,21(3):186-187
[134]余伊君.旋光法测定胃炎合剂中硫酸庆大霉素含量[J].药品检测.2002,11(7):49
[135]姚龙泉,张西臣,陈国庆等.三氮脒、三氮脒脂质体和复方三氮脒注射液在兔体内的药代动力学和组织残留[J].中国寄生虫病防治杂志,2002,15(6):367-369.
[136]陈世琼.猪肠道中抑制大肠杆菌的乳酸菌的分离与筛选[J].中国农业大学学报,2002,7(5):117-120
[137]龙德武,刘传银,赵鸿雁陆光汉.铜(Ⅱ)-三氮唑偶氮络合物与脱氧核糖核酸相互作用的电化学行为[J].分析化学,2002,30(10):1250-1253
[138]祁超,李伟国,张玉静,刘立岩,赵大庆.圆二色谱和紫外可见光谱研究抗体-卟啉的相互作用[J].分析化学,2002,30(5):601-604
[139]王亚俐,王海芳.光谱法研究苯甲酸钠与牛血清白蛋白的作用[J].北京大学学报,2002,38(2):159-163
[140]杨绮华.药物载体系统的研究进展[J].中国新药杂志,2002,11(8):593-596.
[141]王文波,孙丰忠,王勇.新型制剂技术-脂质体研究进展[J].中国制药信息,2002,18(8):23-24
[142]徐福洲,于金洛,杨兵等.抗猪产肠毒素大肠埃希氏菌卵黄抗体的研制及攻毒治疗试验[J].华北农学报,2002,17(4):108-112
[143]吴镭,张强,杨晓青.我国药剂学基础研究的热点领域和基本思路简析[J].中国医药工业杂志,2002,33(2):99.
[144]徐雯,潘俊,陆伟跃.受体介导脂质体的研究进展[J].国外医药-合成约、生化约、制剂分册,2002,23(5):293
[145]徐叔云,卞如廉,陈修.药理学实验方法[M].人民卫生出版社,2002:1647-1670.
[146]张桥.卫生毒理学基础[M].北京:人民卫生出版社,2003:77-84
[147]高云英,赵发苗,张金良等.鸡抗猪大肠杆菌高免卵黄抗体的研制与应用[J].畜牧兽医杂志,2003,22(1):6-8
[148]倪型灏,孙文勇.胃肠道间质瘤临床病理特征的再认识[J].医学临床研究,2003,20(10):773-777
[149]于军,于红,肖洋,等.射干提取液对铜绿假单胞菌PA 11株R质粒体内外消除作用[J].吉林大学学报(医学版),2003,29(2):170-171.
[150]陈祥,高松,王雷等.华东地区致出生仔猪腹泻大肠杆菌的流行病学[J].猪的重要疫病防治研究新成果汇编,2003,10:578-583
[151]蔡家祥.家畜传染病学(第四版)[M].中国农业出版社,2003:46-51
[152]李秀敏.细菌耐药机制研究进展[J],中国新药杂志,2003,12(1):29-31
[153]陈祥,高松,王雷等.华东地区致出生仔猪腹泻大肠杆菌的流行病学[J].猪的重要疫病防治研究新成果汇编,2003,10:578-583
[154]郭长彬 徐志斌 徐志斌.基于HPPK靶标酶的分子对接研究-金属离子Mg2+的影响[J].化学学报,2003,61:1653
[155]王君,任百祥.药物与蛋白相互作用的分析测试[J].中国新医药,2003,2(5):47-48
[156]杨金菊,王继武,李永红,戴平,李荣秀.人白蛋白与3-氨甲基吡啶相互作用的研究[J].中国药学杂志,2003,38(11):882-885
[157]徐铮奎.迅猛发展中的“非常规”制剂市场[J].中国制药信息,2003,19(9):37-38
[158]姜平.兽医生物制品学[M].中国农业出版社,2003.444-50
[159]汪多仁.卵磷脂脂质体开发与应用进展[J].化工中间体,2003(10):23-36
[160]张国红,邓学兵,王刚.动物脂质体技术与产品的研究[J].中国动物保健,2003,8:4-5.
[161]高申,钟延强.现代药物新剂型新技术[M].北京:人民军医出版社,2003,197-219.
[162]陈亮,陈学敏.国内靶向制剂的研究现状[J].儿科药学杂志,2003,9(1):10-12.
[163]胡英.膜融合脂质体给药系统的研究进展[J].国外医学药学分册,2004,31(1):40-43
[164]田景振,张玉娟.脂质体的研究进展[J].齐鲁药事,2004,23(1):40-42
[165]雷肇祖,章健,章健.工业菌种改良述评[J].工业微生物,2004,34(1):39-50
[166]贲百灵,韩华.猪大肠杆菌病的诊断和防治的一点体会[J],畜牧兽医科技信息,2004(9):42
[167]糜祖煌.氨基糖苷类修饰酶及其基因检测[J].现代实用医学,2004,16(1):25-31
[168]纪竹生,刘买利,胡继明.药物配体与生物大分子受体相互作用核磁共振的研究进展[J].分析化学,2004,32(11):1532-1537
[169]王月,肖尚友,陈华,夏之宁.硝基苯胺同牛血清白蛋白的相互作用及其毒性[J].环境化学,2004,23(5):529-534
[170]白炳莲,李敏.基于氢键的自组装超分子体系 化学通报,2004,(2),124
[171]凌保东,余闲,谢勇恩.临床分离产AmpCβ内酰胺酶阴钩肠杆菌的耐药性分析[J].中国抗生素杂志,2004,30(4):50-53
[172]杨俐锋,喻琳,刘艳,周宁,陈勇云,赵玉芬.电喷雾质谱法研究酸和盐诱导下蛋白质的折叠状态的改变[J].厦门大学学报,2004,43(4):518-521
[173]袁彩霞,杨频.二氯邻菲咯啉合钴与DNA的相互作用研究[J].山西大学学报,2004,27(2):163-166
[174]刘雪锋,夏咏梅,方云,邹鲁,刘玲玲.三种香豆素类中药小分子与牛血清白蛋白的相互作用[J].化学学报,2004,62(12):1484-1490
[175]吴会灵,杨苋原,张重杰,何锡文.小分子与核酸相互作用的研究进展[J].分析化学,2004,32(9):1256-1261
[176]吴英才,袁一方,徐艳平.表面等离子共振传感器的研究进展[J].传感器技术,2004,23(5):1-5
[177]邬春华,吕元琦,袁倬斌.毛细管电泳法测定沙棘药物制品心达康片中的山奈酚和槲皮素[J].分析测试学报,2004,23(3):73-75
[178]袁耀武,张伟,马雯,等.大肠埃希菌O157菌体抗原免疫血清的制备及简易纯化.中国卫生检验杂志,2004(1):98-99
[179]刘永春,胡之德.药物与蛋白质相互作用的荧光光谱法研究概述[J].宝鸡文理学院学报。2005,25(1):42-49
[180]张朝红,臧树良,耿兵等.紫外和圆二色光谱法研究丁基锡化合物与牛血清白蛋白的相 互作用[J].分析科学学报,2005,21(2):179-181
[181]张瑞萍,再帕尔·阿不力孜.分子间非共价相互作用的质谱分析方法研究进展[J].分析测试学报,2005,24(1):117-122
[182]邹其森,刘延霞.肺癌靶向治疗药物进展[J].世界临床药物,2005,26(5):283
[183]王自然.中药口服液治疗人工感染仔猪大肠杆菌试验研究[J].中兽医医药杂志,2005,4:18-20
[184]猪大肠杆菌病及其防治[J].疫病防制.2005.11:31-32
[185]钟江国,王卓南.鸡场大肠杆菌耐药性调查与对策[J].中国兽医杂志,2005,22(5):38-39
[186]王秀敏 邓英杰 赵春杰等.β-榄香烯长循环脂质体的研究.中华医药杂志,2005,5(3):200-201
[187]郭伶,陈宝利,杨松.锦州地区猪大肠杆菌病流行病学调查[J].畜牧兽医科技信息,2006,11(5):26
[188]韩英.炎症性肠病生物疗法新进展[J].中华内科杂志.2006.45(3):253-255
[189]陈宇洲,王东凯.炎症性肠病的结肠生理特征和结肠定位给药[J].中国药学杂志.2006.41(2):86-90
[190]赵耘,沈青春,沈润爱.致肾脏、肠道病变大肠杆菌的分离及生物特性鉴定[J].中国预防兽医学报,2006,28(1):51-54
[191]邵海枫,赵晓智,王卫萍,等.肺炎克雷伯菌中质粒介导的AmpC酶及相关表型研究[J].中华检验医学杂志,2006,29(2):117-119
[192]范义湘,罗荣城,方永鑫,严晓,吕成伟.干扰素-γ对乳腺癌细胞Her2/neu表达及131I-Herceptin抑制肿瘤细胞增殖的影响[J].癌症.2006.25(4):443-446
[193]张奇,柳时,周小菊,项光亚.叶酸靶向的给药系统[J].中国药学杂志.200.4(1):1-4
[194]王玮琴,周慧君,陈耀.青蒿琥酯抑制鸡胚尿囊膜血管生成作用的研究[J].中国药学杂志.2006,41(1):22-24
[2]Rahman YE,Cerny EA,Tollaksen SL,et al.Liposome-encapsulated actinoycin D:potential in cancer chemotherapy.Proc Soc Exp BiolMed.1974,146(4):1173-1176
[3]Fouda HG.Determination of diminazene in plasma by high-perforarnnce liquid chromatography[J].Journal of Chromatographic Science,1977,15:537-538
[4]L.Meyer Jones,Nicholas H.Booth,Leslie E.McDonald主编.兽医药理学与治疗学[M],冯淇辉,中葆和,戎耀方,等泽.上海科学文献出版社,1982:727
[5]Tsutumi F.Sakakibara M,Oshima N.Structure and dynamicproperties of solution SBR coupled with tin compounds[J].Rubber Chem istry and Technology.1990,69(1):8
[6]Alzheimer disease:evidence for chromosome 19 linkage[J].Ame J Hum Genet,1991,48(5):1034
[7]Hall RM,brookesDE,stokesHW.Site-specific Insertion of Genes into Integrons:Role of the 59-base Element and Determina-tion of the Recombination Cross-over point[J].Mol Microbiol,1991,5:1941-1953
[8]Murray BE.New aspects of antimicrobial resistance and resulting the therapeutic dilem mas[J]InfectDis,1991,163:1185
[9]Fattal E,Rojas J,Youssef M,et al.Liposome-entrapped ampicilin in the treatment of experimental murine listeriosis and salmonellosis[J].Antimicrob Agents Chemother,1991,35(4):770
[10]Tomioka H,Saito H,Sato K,et al.Therapeutic efficacy of liposome-encapsulated kanamycinagainst mycobaterium intracellulare infection induced in mice[J].Am Rev Respir Dis,1991,144(3):575
[11]Yeh MY.R(,filer SR.Doxorubicin:monoclonal antibody conjugate for therapy of human cervical carcinomal[J].lnt J Cancer.1992,51(2),274-282
[12]Woodle MC,Lasic DD.Sterically stabilized liposomes[J].Biochim Biophys Acta,1992,1113(2):171-199
[13]Ana cabanes,Margarita arboix,Jose M,et al.Pharmacoki-netics of enrofloxacin after intravenous and intramuscular in-jection in rabbits[J].Am J Vet Res,1992,53(11):2090-2093
[14] Mehta RT, Kevhani A, Mequeen T.I, et al. In vitro activities of free and liposomal drugs against Mvcobacterium avium-M, intracellulare complex and M. tuberculonis[J]. Antimicrob Agents Chemother,1993,37(12):2584
[15]Pastenack R F, BustamanteC, Collings P J. Am. Chemv.Soci, 1993, 115: 5393-5399
[16] Forssen EA, Ross ME. Daunoxome treatment of solid turners :preclinical and clinical investigations[J]. J Liposome Res , 1994 ,4 (1): 481—512
[17] Yuan F, Dellian M, FukumuraD, et al. Vascular permeability in human tuman tumor xenograft molecular size dependence and cutoff size [J].Cancer Res, 1995. 55: 3752—3756.
[18]SakaeU, KazuoM, Osamu I, et al.Enhanced tumor targeting and improved antitumor activity of doxorubi by cin long-circulating liposomes containing amphipathic poly(ethylene glycol)[J]. International Journal of Pharmaceutics, 1995, 126: 41 —48
[19] Zhu Z, Kralovec J Inhibition of epstein—Barr—virus—transformed human chornic lymphocytic leukaemic B cells with monoclonal antibody adriamycin (doxorubicin) conjugates [J]. Cancer Immunol Immunother, 1995, 40(4): 257—267
[20] Hsiech H L, Quirk R P. Anionic polymerization: principles and practical applications[J]. New York: Marcel Dekker Inc. 1996. 447
[21] Mendelsohn L, Epidermal growth factor receptor inhibition by a monoclonal antibody as antibody therapy[J] Clin( ncer Res, 1997, 3(12): 2703—2707
[22] Pardue R L, White C A. Pharmacokinetic evaluation of liposomal encapsulated ampicillin in male and female rats[J]. Biopharm Drug Dispos 1997,18(4):279-292.
[23] Vladimir A,Slepushkin, Sengio S,et al.Sterically stabilized pH-sensitive liposomes[J]. Journ Biol Chemi, 1997, 272: 2382-2388
[24] Pharmacokinetic Pardue R L, White C A. Pharmacokinetic evaluation of liposomal encapsulated ampicillin in male and female rats[J].Biopharm Drug Dispos, 1997,18(4):279—292.
[25] Wiechens B , Krausse R , Crammeer J B , et al. Clearance of liposome-incorporated ciprolloxacin after intravitreal injection in rabbit eyes[J]. Klin Monatsb Augenheilkd , 1998, 213(5):284-292.
[26] Omelyanenko V, Kopekuaa P Targetable HPMA copolymer-adriamycin conjugates, recognition, internalisation and subcellular fate[J]. J Controlled Release, 1998, 53(1—3): 25— 37.
[27] Reiter Y. Pastan 1 Recombinant Fv immunotoxins and Fv flag—ments as nox el agents for cancer therapy and diagnosis[J] J rends Biote～h, 1998, 16(12): 513—520
[28] Oussoren C, Velinova M, Scherphof G. et al. Lymphatic uptake and biodistribution of liposomes after subcutaneous injection. IV. Fate of liposomes in regional lymph nodes[J]. Biochim Biophys Acta, 1998, 1370: 259-272.
[20] Higuchi M, Fokin A, Masters TN, et al. Transport of colloidal particles in lymphatics and vasculature after subcutaneous injection[J]. J Appl Physiol, 1999. 86: 1381 — 1387
[30] Brocchini S, Duncma R Polymer drIJg conjugates: drug release from pendent linker: A Mathiowitz E(ED)Encyclopaedia of controlled drug delivery[M]. New York: Wiley, 1999: 786-816
[31] Takahashi H, Adacchi K. Experimental treatment of malignant gliomas with human monoclonal antibody drug conjugates[J]. AnticancerRes, 1999, 19(12): 4151—4156
[32] Ota Y, Fukasawa I Antitumor effect of monoc lonal antibody-car-boplatin conjugates in nude mice bearing ovarian cancer cells[J]. Int J Clin Oncol, 1999, 4(2): 236—240
[33] Ochs MM, Mc Cusker MP, Bains M, et al. Negative regulation of the Pseudomonas aeruginosa outer membrane porin OprD selective for imipenem and basic aminoacids [J]. Antimicrob Agents Chemother,1999, 43:1085—1090.
[34] Mckessar SJ, Berry AM,Bell JM, et al. Genetic Characterization of VanG,A novel vancomycin reisistance locus of Enterococcus fecalis [J] ,Antimicrob Agents Chemother,2000,44:3224— 3228.
[35] O samu I, Kazuo M, Hironobu Y, et al.Targeting chemotherapy to solid tumors with long-circulation thermosensitive liposomes and local hypertherthermia[J]. Jpn J Cancer Res, 2000, 91: 118-126
[36] Duncan R, Gac-Breton S. Keane R, et al Polymer-drug conjugates, PI)EI l and PELT: basic principles for design and transfer from the la}× )ratory to clinic[J]. J Controlled Release. 2001, 74(1—3)135 -146
[37] Garett MC. Iargeted drug conjugates: principles and progress [J].Adv Drug Deltev Rev, 2001, 53(2): 171-216
[38] Liebe V, Bruckmann M, Fischer KG, et al. Biologicalrelevance of anti recombinant hirudin antibodies-results from nvitro and in vivo studies[J]. Semin Thromb Hemost, 2002, 28(5): 483-490
[39] Phillips RK, Wallace MH, Lynch PM, et al. A randomized, double blind, placebo controlled study of celecoxib, a selective cyclooxygenase 2 inhibitor, on duodenal polyposis in familial adenomatous polyposis[J]. Gut, 2002, 50(6): 857-860
[40] Garberk, TeBexxar.FDA weeting offers long—awaitedchance at approvat[J].J Natl Cancer Inst. 2002, 94 (23) : 1738.
[41] Richardson PG, Barlogie B, Berenson MD, et al. A phase 2 study of bortezomib in relapsed, refractory myeloma[J]. N Engl J Med, 2003, 348(26): 2609-2617
[42] wusu-Asiedu A. et al. Response of early-weaned pigs to an enterotoxigenic Escherichia coli(K88) challenge when fed diets containing spray-dried porcine plasma or pea protein isolate plus egg yolk antibody [J] .J.Anim. Sci,2003,82: 1781-1789
[43] Nakata E,Hunter N,Mason K,et al.C225 antiepidermal growth factor receptor antibody enhances the efficacy of docetaxel chemoradiotherapy[J].Int J Radiat Oncol Biol Phys,2004,59(4): 1163 — 1173
[44] Martin Tulp, Lars Bohlin. Unconventional natural sources for future drug discovery [J]. Drug Discovery Today,2004,9( 10):9
[45] Mire-sluisa AR, Barrettb WC, Devanarayan V, etal. Recommendations for the design and optimization of immunoassaysused in the detection of host antibodies against biotechnol—ogy products[J]. J Immunol Methods, 2004, 289(1): 1 —16.
[46] Hurvitz H, Fehrenbacher L, NovotnyW, et al. Bevacizumab plus iinotecan, fluoruracil, and leucovorin for metasratic correctal[J].Nev Engl J Med.2004, 350 (23) : 2335
[47] Dittmann K,Mayer C,Fehrenbacher B,et al.Radiation-induced epidermal growth factor receptor nuclear import is linked to activation of DNA-dependent protein kinase.J Biol Chem,2005,280 (35):31182-31189
[48] Wadhwa M, Gaines-das R. Thorpe R, etal. Detection, measurement and characterization of unwanted antibodies inducedby therapeutic biologicals[J]. Dev Biol(Basel), 2005, 122: 155— 170.
[49] Shen B, Nonaka K, Nonaka K. Enediyne natural product: Biosynthesis and prospect towards engineering novel antitumor agents [J]. Bentham Science Publishers,2005,2 : 357-369
[50] Wan, Yinhua; Ghosh, Raja, Cui; Zhanfeng and Hale, Geoff. Fractionation of BSA and Monoclonal Antibody Alemtuzumab Using Carrier Phase Ultrafiltration [J]. Biotechnol Bioeng, 2005,90(3):303-315
[51] Feng Q, Yang J, Luo P,et al. LT(K63/R72),a new mutant of Escherichia coli heat-labile enterotoxin,exhibits characteristics more similar to LT(K63)than LT(R72)[J].Acta Biochim Biophys Sin(Shanghai),2005.37(2):126-132
[52]Pao W,Miller VA.Fpidermal growth factor receptor mutations,small-molecule kinase inhibitors,and non-smal1-cell-lung cancer:current knowledge and future directions.J Clin Oncol,2005,23(11):2556-2568.
[53]Burtness B.The role of cetuximab in the treatment of squamous cell cancer of the head and neck.Expert Opin Biol Ther,2005,5(8):1085-1093.
[54]Raizer JJ.HER1/EGFR tyrosine kinase inhibitors for the treatment of glioblastoma multiforme.J Neurooncol,2005,74(1):77-86.
[55]Harari PM,Huang S.Radiation combined with EGFR signal inhibitors:head and neck cancer focus.Semin Radiat Oncol,2006,16(1):38-44
[56]Tillmann HC,Kuhn B,Kranzlin B,et at.Efficacy andimmunogenicity of novel erythropoietic agents and conventionalrhEPO in rats with renal insufficiency[J].Kidney lnt,2006,69(1):60-67.
[57]Chang W Y.Carter R,Hardy T,etal.Inhibition of Escherichia coil heat-labile enterotoxin B subunit pentamer(EtxB5)assdmbly in vitro using monouonal antibodies[J].Biol chem.,2006,10:1074
[58]T.Luke Simmons and Kerry L.McPhail etc.Belamide A=Belamide A,a new antimitotic tetrapeptide from a Panamanian marine cyano- bacterium[J].Tetrahedron Letters,2006,47(20):3387-3390.
[59]Wilson GD,Bentzen SM,Harari PM.Biologic basis for combining drugs with radiation.Semin Radiat Oncol,2006,16(1):2-9
[60]施新猷.休克模型的复制[J].医学动物实验方法,1980,250-254
[61]胡远峰.大鼠糖尿病模型[J].中华内科杂志,1981,(9):535
[62]卢兴.儿种常用心律失常动物模型的复制[J].人类疾病的动物模型,1982,1(19):182-199
[63]重尔昌.大白鼠的实验性巨结肠[J].人类疾病的动物,1982,271(274):308-310
[64]吴荣庆.大白鼠胆管炎模型制作方法的介绍[J],武汉医学院学报,1983,(4):364
[65]夏家明.大鼠实验性急性肺水肿模型[J].上海实验动物科学,1985,5(4):240
[66]吴若铢.天花粉引起被动肺过敏的大鼠模型[J].中国药理学报,1985,6(1):68
[67]谢明智.实验性肥胖及糖尿病大鼠模型[J].药学学报,1985,20(11):801
[68]郭尧君.分光光度技术及其在生物化学中的应用[M].科学出版社,1987.230-231
[69]章文贡,陈田安,陈文定.铝酸酯偶联剂改性碳酸钙的性能与应用[J].中国塑料,1988,2(1):3
[70]Habamnh CM等.丁胺卡那霉素的特点平临床应用[J].国外医药抗生索分册,1989,10(5):365-368.
[71]何晓青.卫生防疫细菌检验[M].新华出版社1989 216-520
[72]容健材,廖锡麟.新编药理学[M].江苏科技出版社,1990:515
[73]傅永林.偶联剂住塑料复合材料中的应用[J].中国塑料.1991.5(3):20
[75]李战强.王永沛,李荣兴.等.双金属偶联剂FPM在电缆橡胶中的应用[J].无机盐工业.1992.120(5):40
[76]肖希龙,沈建忠,朱蓓蕾等.泰乐加和呋喃它酮对大肠杆菌与沙门氏菌的药效研究[J]畜牧兽医学报,1993,24(1):81-86
[77]王树槐,潘松年.盐酸环丙沙星注射液对仔猪大肠杆菌病的临床药效试验[J].中国兽药杂志,1994,28(3):34-35、
[78]徐美娟,聂春发等.临床常用氨基糖苷在烫伤兔体中的药动学改变[J].中国抗生素杂志,1994,19(1):42-45.
[79]张仰亨,张榕平.比色测定法测定硫酸庆大霉素及其注射液的含量[J].海峡药学,1994,6(3):27-28.
[80]何笑蓉。赵研.傅得兴.一阶导数光谱法测定硫酸庆大霉素制剂的含量[J].中国医院约学杂志。1995,15:(11):505-507
[81]李好枝,高光恩,田洁.荧光光谱法测定硫酸庆大霉素注射液的含量[J].西北药学杂志,1995,10(1):5-7
[82]李铜铃,庞其捷,贺于玲,等.肝靶向抗疟药半乳糖基似糖白蛋白-伯氨喹偶联物和磷酸伯氨喹的药代动力学[J].药学学报,1995,30(10):721-725
[83]李富荣,王琪,李守唐.吡喹酮和吡喹酮脂质体RP-HPLC法测定及药代动力学研究[J].中国药理学通报,1995,11(4):341-343
[84]陈士朝.轮胎的性能要求与合成橡胶_的发展[J].合成橡胶工业.1995.18(5):260
[85]张强,廖工铁,寿旦.用高效液相色谱法测定兔血清中硫酸庆大霉素的含量[J]中国医院药学杂志,1995,15(4):147-149.
[86]杨秀岑,伍莉萍。邓安平等.生物素标记增强化学发光法测定血清中庆大霉素含量[J].华西医大学报,1995,26(2):234-236
[87]张永恒.现代靶向制剂[M].黑龙江科学技术出版社,1995:1.
[88]丁鼎乐.靶向制剂国内外研究进展[J].中国医药情报,1996,2(5):301
[89]张强,廖工铁,魏大鹏等.用毫微球增加体外小鼠腹腔巨噬细胞与大鼠肝细胞对庆大霉素的摄取[J].药学学报,1996,31(5):375-380
[90]徐葛林,严家信.脂质体用作疫苗佐剂的研究进展[J].国外医学-预防、诊断、治疗用生物制品分册,1996,19(1):10.
[91]崔春英。席德荣.胃乐合剂中硫酸庆大霉素的含量测定.中国药事.1996。10(2):117.
[92]武德珍,白宗武.稀土偶联剂处理碳酸钙填充PVC复合材料及其性能的研究[J].现代塑料加工应用.1996,8(1):10.
[93]鲍爱华.世界溶聚丁苯橡胶的主要技术进展[J].石油化工动态,1996.4(3):18
[94]张开.高分子界面科学[M].中国石化出版社.1997.223
[95]陈士朝.溶聚丁苯橡胶的技术进展[J].合成橡胶工业.1997.20(1):6
[96]徐宝梁,李谨,马颖.有关杯碟法沁定壮观霉素灵敏度的影响因素[J].微生物学通报,1997,24(6):372-375
[97]王晶钰.陕西省鸡致病性大肠杆菌生物学特征和免疫防治研究[J].西北农林科技大学学报,1997,(2):97-101.
[98]廖工铁.靶向给药制剂[M].成都:四川科学技术出版社,1997:334-371.
[99]平其能.现代药剂学[M].中国医药科技出版社,1998:588-621.
[100]陆彬.药物新剂型与新技术[M].人民卫生出版社,1998:32,
[101]姜为东,黄引贤.广东省仔猪腹泻大肠杆菌病血清型鉴定[J].中国兽医学报,1998,18(1):63-65.
[102]张均田.现代药理实验方法[M].北京:北京医科大学中国协和医科大学联合出版社,1998:1412-1415
[103]刘继兰,李敬玺,袁有申等.“9381-杀灭剂”对仔猪大肠杆菌病的治疗试验[J].中国畜禽传染病,1998,20,(1):49-50
[104]顾觉奋,王鲁燕,张俊琪.阿米卡星脂质体在小鼠体内的组织分布[J].中国抗生素杂志,1998,23(4):294-297
[105]陆彬.药物新剂型与新技术[M].人民卫生出版社,1998:326-329
[106]梁亮,廖列文,崔英德.铝一锆有机金属络合物偶联剂的合成及应用[J].精细化工,1999.16(4):4
[107]高崧,刘秀梵,张如宽.我国部分地区禽病源性大肠杆菌的分离和鉴定[J].畜牧兽医学报,1999,30:164-171.
[108]王庆民,宋尚龙.用旋光法测定硫酸庆大霉素注射液的含量[J].药品检测,1999,8(7):45-46.
[109]舒泉.细菌DNA分析在流行病学中的应用[J].中华流行病学杂志,1999,(12):52-54
[110]张秀英.抗生素的耐药机制分类及概述[M].1999年全国兽医学会微生物学学会年会论文集,205-209
[111]何宁,王昌禄,顾晓波等.杯碟法检测乳酸菌素活性优化条件的研究[J].天津轻工业学院学报,1999,2:18-20
[112]甑永苏.单克隆抗体药物治疗肿瘤的研究现状及展望[J]中国医学科学院学报,2000,2(1):9-13
[113]江寅生,张皖华.庆大霉素滴鼻剂的研制及含量测定[J]中国药业,2000,9(10):53
[114]吴建良,沈特庞,陈晓萍.电荷转移络合--阶导数光谱法测定硫酸庆大霉素滴眼液的含最[J].西北药学杂志,2000.15(4):152-153
[115]朱立平,陈学清.免疫学常用实验技术[M].人民军医出版社,2000:57-61
[116]张永泉。杜战江,刘云等.d,JL硫酸庆大霉素颗粒含量测定方法研究[J].中国药事,2000,14(3):177-178.
[117]庞扶民。李静,张汉宁.用旋光法测定硫酸庆大霉素溶液的含量[J].中国煤炭工业医学杂志,2000.3(2):151
[118]姜为东,黄引贤.广东省仔猪腹泻大肠杆菌病血清型鉴定[J].中国兽医学报,2000,18(1):63-65
[119]刘又宁,王睿,邓宝军等.头孢呋辛钠治疗细菌性感染临床评价[J].中国临床药理字杂志,2000,16(4):251-254
[120]高德伟 王华琼 张进川 刘春雷.第四代头孢菌素头孢吡肟在老年肺炎治疗中的应用[J].中华医院感染学杂志,2000,10(3):221-222
[121]吕志堂 刘志恒 刘志恒.一种从土壤样品中选择性分离假诺卡氏菌的方法[J].微生物学报,2000,40(4):406-113
[122]鞠洪涛.大肠杆菌耐药基因定位及耐药质粒的消除[J].中国兽医学报,2000(6):2
[123]刘业兵,高松.禽病原性大肠杆菌Ⅰ型茵毛单克隆抗体的研制极其对分离株的检测[J].中国兽医学报,2000,20(2):148-151
[124]赵雨,徐金泽,赵大庆.二维拉曼相关光谱研究稀土离子对血红蛋白结构的影响[J].高等学校化学学报,2000,21(12):1913-1915
[125]俞英,程晓伟.牛血清白蛋白与酸性品红的结合反应[J].分析化学,2000,28(5):583- 586
[124]赵坤,王三虎.克痢注射液对仔猪大肠杆菌病治疗效果研究[J].中国医学杂志,2000,(3):8-9
[125]于波涛,张志荣,曾仁杰.约质体-一种新型给药系统[J].中国药学杂志,2000,35(11):721.
[126]彭应旭,廖工铁.骨髓靶向给药制剂的研究进展[J].中国药学杂志,2000,35(2):73
[127]张静,丁永红.木质素作为偶联剂在橡胶中的作用[J].特种橡胶制品,2001,22(6):22
[128]包晓玉,陈建国,陈欣,党元林.芦丁与DNA相互作用的电化学研究[J],分析试验室,2001,20(2):1-3
[129]凌连生,杨洗,何治柯,曾云鹗.小分子与DNA相互作用的光谱研究[J].分析科学学报,2001,17(1):11-15
[130]范盛先,袁宗辉,孔科等.庆大霉素在猪肉和鸡肉组织中残留的微生物学检测方法研究[J].华中农业大学学报,2001:557-560
[131]杨晓青,张强,吴霖.我国药剂学靶向制剂研究的基本思路[J].中国药学杂志,2001,36(12):797
[132]夏业才,姚文生,刘惠文.沙拉沙星注射液对人工诱发仔猪大肠杆菌病的临床疗效试验[J].中国兽药杂志,2001,35(1):66-67
[133]黄玲.胃宁冲剂中盐酸酱鲁卡因与硫酸庆大霉素的含量测定[J].中国医院药学杂志,2001,21(3):186-187
[134]余伊君.旋光法测定胃炎合剂中硫酸庆大霉素含量[J].药品检测.2002,11(7):49
[135]姚龙泉,张西臣,陈国庆等.三氮脒、三氮脒脂质体和复方三氮脒注射液在兔体内的药代动力学和组织残留[J].中国寄生虫病防治杂志,2002,15(6):367-369.
[136]陈世琼.猪肠道中抑制大肠杆菌的乳酸菌的分离与筛选[J].中国农业大学学报,2002,7(5):117-120
[137]龙德武,刘传银,赵鸿雁陆光汉.铜(Ⅱ)-三氮唑偶氮络合物与脱氧核糖核酸相互作用的电化学行为[J].分析化学,2002,30(10):1250-1253
[138]祁超,李伟国,张玉静,刘立岩,赵大庆.圆二色谱和紫外可见光谱研究抗体-卟啉的相互作用[J].分析化学,2002,30(5):601-604
[139]王亚俐,王海芳.光谱法研究苯甲酸钠与牛血清白蛋白的作用[J].北京大学学报,2002,38(2):159-163
[140]杨绮华.药物载体系统的研究进展[J].中国新药杂志,2002,11(8):593-596.
[141]王文波,孙丰忠,王勇.新型制剂技术-脂质体研究进展[J].中国制药信息,2002,18(8):23-24
[142]徐福洲,于金洛,杨兵等.抗猪产肠毒素大肠埃希氏菌卵黄抗体的研制及攻毒治疗试验[J].华北农学报,2002,17(4):108-112
[143]吴镭,张强,杨晓青.我国药剂学基础研究的热点领域和基本思路简析[J].中国医药工业杂志,2002,33(2):99.
[144]徐雯,潘俊,陆伟跃.受体介导脂质体的研究进展[J].国外医药-合成约、生化约、制剂分册,2002,23(5):293
[145]徐叔云,卞如廉,陈修.药理学实验方法[M].人民卫生出版社,2002:1647-1670.
[146]张桥.卫生毒理学基础[M].北京:人民卫生出版社,2003:77-84
[147]高云英,赵发苗,张金良等.鸡抗猪大肠杆菌高免卵黄抗体的研制与应用[J].畜牧兽医杂志,2003,22(1):6-8
[148]倪型灏,孙文勇.胃肠道间质瘤临床病理特征的再认识[J].医学临床研究,2003,20(10):773-777
[149]于军,于红,肖洋,等.射干提取液对铜绿假单胞菌PA 11株R质粒体内外消除作用[J].吉林大学学报(医学版),2003,29(2):170-171.
[150]陈祥,高松,王雷等.华东地区致出生仔猪腹泻大肠杆菌的流行病学[J].猪的重要疫病防治研究新成果汇编,2003,10:578-583
[151]蔡家祥.家畜传染病学(第四版)[M].中国农业出版社,2003:46-51
[152]李秀敏.细菌耐药机制研究进展[J],中国新药杂志,2003,12(1):29-31
[153]陈祥,高松,王雷等.华东地区致出生仔猪腹泻大肠杆菌的流行病学[J].猪的重要疫病防治研究新成果汇编,2003,10:578-583
[154]郭长彬 徐志斌 徐志斌.基于HPPK靶标酶的分子对接研究-金属离子Mg2+的影响[J].化学学报,2003,61:1653
[155]王君,任百祥.药物与蛋白相互作用的分析测试[J].中国新医药,2003,2(5):47-48
[156]杨金菊,王继武,李永红,戴平,李荣秀.人白蛋白与3-氨甲基吡啶相互作用的研究[J].中国药学杂志,2003,38(11):882-885
[157]徐铮奎.迅猛发展中的“非常规”制剂市场[J].中国制药信息,2003,19(9):37-38
[158]姜平.兽医生物制品学[M].中国农业出版社,2003.444-50
[159]汪多仁.卵磷脂脂质体开发与应用进展[J].化工中间体,2003(10):23-36
[160]张国红,邓学兵,王刚.动物脂质体技术与产品的研究[J].中国动物保健,2003,8:4-5.
[161]高申,钟延强.现代药物新剂型新技术[M].北京:人民军医出版社,2003,197-219.
[162]陈亮,陈学敏.国内靶向制剂的研究现状[J].儿科药学杂志,2003,9(1):10-12.
[163]胡英.膜融合脂质体给药系统的研究进展[J].国外医学药学分册,2004,31(1):40-43
[164]田景振,张玉娟.脂质体的研究进展[J].齐鲁药事,2004,23(1):40-42
[165]雷肇祖,章健,章健.工业菌种改良述评[J].工业微生物,2004,34(1):39-50
[166]贲百灵,韩华.猪大肠杆菌病的诊断和防治的一点体会[J],畜牧兽医科技信息,2004(9):42
[167]糜祖煌.氨基糖苷类修饰酶及其基因检测[J].现代实用医学,2004,16(1):25-31
[168]纪竹生,刘买利,胡继明.药物配体与生物大分子受体相互作用核磁共振的研究进展[J].分析化学,2004,32(11):1532-1537
[169]王月,肖尚友,陈华,夏之宁.硝基苯胺同牛血清白蛋白的相互作用及其毒性[J].环境化学,2004,23(5):529-534
[170]白炳莲,李敏.基于氢键的自组装超分子体系 化学通报,2004,(2),124
[171]凌保东,余闲,谢勇恩.临床分离产AmpCβ内酰胺酶阴钩肠杆菌的耐药性分析[J].中国抗生素杂志,2004,30(4):50-53
[172]杨俐锋,喻琳,刘艳,周宁,陈勇云,赵玉芬.电喷雾质谱法研究酸和盐诱导下蛋白质的折叠状态的改变[J].厦门大学学报,2004,43(4):518-521
[173]袁彩霞,杨频.二氯邻菲咯啉合钴与DNA的相互作用研究[J].山西大学学报,2004,27(2):163-166
[174]刘雪锋,夏咏梅,方云,邹鲁,刘玲玲.三种香豆素类中药小分子与牛血清白蛋白的相互作用[J].化学学报,2004,62(12):1484-1490
[175]吴会灵,杨苋原,张重杰,何锡文.小分子与核酸相互作用的研究进展[J].分析化学,2004,32(9):1256-1261
[176]吴英才,袁一方,徐艳平.表面等离子共振传感器的研究进展[J].传感器技术,2004,23(5):1-5
[177]邬春华,吕元琦,袁倬斌.毛细管电泳法测定沙棘药物制品心达康片中的山奈酚和槲皮素[J].分析测试学报,2004,23(3):73-75
[178]袁耀武,张伟,马雯,等.大肠埃希菌O157菌体抗原免疫血清的制备及简易纯化.中国卫生检验杂志,2004(1):98-99
[179]刘永春,胡之德.药物与蛋白质相互作用的荧光光谱法研究概述[J].宝鸡文理学院学报。2005,25(1):42-49
[180]张朝红,臧树良,耿兵等.紫外和圆二色光谱法研究丁基锡化合物与牛血清白蛋白的相 互作用[J].分析科学学报,2005,21(2):179-181
[181]张瑞萍,再帕尔·阿不力孜.分子间非共价相互作用的质谱分析方法研究进展[J].分析测试学报,2005,24(1):117-122
[182]邹其森,刘延霞.肺癌靶向治疗药物进展[J].世界临床药物,2005,26(5):283
[183]王自然.中药口服液治疗人工感染仔猪大肠杆菌试验研究[J].中兽医医药杂志,2005,4:18-20
[184]猪大肠杆菌病及其防治[J].疫病防制.2005.11:31-32
[185]钟江国,王卓南.鸡场大肠杆菌耐药性调查与对策[J].中国兽医杂志,2005,22(5):38-39
[186]王秀敏 邓英杰 赵春杰等.β-榄香烯长循环脂质体的研究.中华医药杂志,2005,5(3):200-201
[187]郭伶,陈宝利,杨松.锦州地区猪大肠杆菌病流行病学调查[J].畜牧兽医科技信息,2006,11(5):26
[188]韩英.炎症性肠病生物疗法新进展[J].中华内科杂志.2006.45(3):253-255
[189]陈宇洲,王东凯.炎症性肠病的结肠生理特征和结肠定位给药[J].中国药学杂志.2006.41(2):86-90
[190]赵耘,沈青春,沈润爱.致肾脏、肠道病变大肠杆菌的分离及生物特性鉴定[J].中国预防兽医学报,2006,28(1):51-54
[191]邵海枫,赵晓智,王卫萍,等.肺炎克雷伯菌中质粒介导的AmpC酶及相关表型研究[J].中华检验医学杂志,2006,29(2):117-119
[192]范义湘,罗荣城,方永鑫,严晓,吕成伟.干扰素-γ对乳腺癌细胞Her2/neu表达及131I-Herceptin抑制肿瘤细胞增殖的影响[J].癌症.2006.25(4):443-446
[193]张奇,柳时,周小菊,项光亚.叶酸靶向的给药系统[J].中国药学杂志.200.4(1):1-4
[194]王玮琴,周慧君,陈耀.青蒿琥酯抑制鸡胚尿囊膜血管生成作用的研究[J].中国药学杂志.2006,41(1):22-24