维氏气单胞菌TH0426胞外产物酶活性及免疫原性分析

为研究维氏气单胞菌(Aeromonasveronii)胞外产物的合成及其功能,本试验采用超滤浓缩法提取Aeromonasveronii TH0426的胞外产物,通过平板琼脂扩散法对其主要酶成分进行初步研究,对蛋白酶、脂酶、卵磷脂酶、淀粉酶、脲酶、明胶酶和溶血活性进行测定,对斑马鱼腹腔注射胞外产物测定其致病性,并对其免疫原性进行初步分析。结果显示,菌株胞外产物具有脂酶、淀粉酶、蛋白酶和溶血活性,人工感染试验结果表明,菌株胞外产物对斑马鱼具有致病性,其半数致死量为3.355 0μg/条。SDS-PAGE表明,ECP由19条蛋白带组成,利用鲤鱼抗ECP血清进行Western blot显示,组成ECP的19条蛋白带中有5条具有免疫原性,能够引发机体的免疫应答反应产生抗体,其相对分子质量为别为110 000,47 000,45 000,42 000和38 000,为Aeromonasveronii胞外产物亚单位疫苗的研制与开发提供理论依据。     

棉籽粕源发酵蛋白质饲料的代谢产物研究

本试验旨在通过液相色谱串联质谱(LC-MS)代谢组学分析平台对不同微生物棉籽粕源发酵蛋白质饲料代谢产物进行研究.采用LC-MS法对对照组、假丝酵母组、酿酒酵母组、复合组的发酵代谢产物进行分析,结合偏最小二乘法-判别分析(PLS-DA)方法对数据进行模式识别和小分子差异代谢产物寻找.结果表明:各试验组代谢产物相对含量与对照组相比差异显著(P＜0.05),且在PLS-DA分析中得到了很好的分离,主要表现在甘露醇、琥珀酸等糖类代谢产物;磷酸胆碱、L-肉碱、甘油磷酸、磷脂酰乙醇胺(PE)和磷脂酰胆碱(PC)等脂类代谢产物;二肽、三肽、甜菜醛、同型半胱氨酸等蛋白质与氨基酸的代谢产物;以及烟酸、阿魏酸、尿嘧啶、乙醇醛等其他途径代谢产物.除与脂类代谢有关的甘油磷酸、PE、PC相对含量显著降低之外(P＜0.05),其他试验组小分子代谢产物相对含量均比对照组显著增加(P＜0.05).由此可见,不同菌种发酵饲料中含有大量糖类、脂类、蛋白质等代谢途径产生的小分子代谢产物,含量因发酵菌种的不同而不同.     

致水貂出血性肺炎铜绿假单胞菌胞外产物生物活性及免疫保护力分析

胞外产物在细菌致病过程中发挥重要作用,为探索胞外产物在铜绿假单胞菌致病过程中的作用,首先从铜绿假单胞菌培养液提取其出其胞外产物,利用小鼠模型分析其致病性,并通过细胞证实了该产物对巨噬细胞功能的抑制作用,进一步测定其胞外产物的酶活性并利用LC-MSMS方法对铜绿假单胞菌胞外产物蛋白成分和功能进行分析。结果表明,铜绿假单胞菌胞外产物对小鼠有致病性,能够显著抑制巨噬细胞吞噬细菌活性,其具有淀粉酶、脂肪酶、蛋白酶、卵磷脂酶活性和溶血活性可能在这一过程中发挥关键作用。LC-MSMS分析鉴定所提纯的胞外产物中有153种蛋白成分,涉及多种酶类、细胞结构蛋白和功能性蛋白。用低剂量胞外产物免疫动物对于铜绿假单胞菌感染具有较明显的保护率,可达90%。本试验为揭示铜绿假单胞菌致病机理,研制新型疫苗奠定基础。     

《天然产物化学》教学方法探讨

《天然产物化学》是化学、化工、生物技术、生物工程、食品科学与工程、制药工程和药学等专业本科生和研究生的一门重要课程,是一门集基础与应用于一体的课程。为进一步提高学生对《天然产物化学》的学习效果,本文对天然产物化学理论课教学方面和实验课教学方面的教学方法进行了分析和总结,以提高其学习效果。     

乳酸菌体外抑菌试验报告

对乳酸杆菌(LL)、嗜酸杆菌(LA)、乳酸链球菌(SL)的抑菌特性进行了初步探讨。采用牛津杯法对3株乳酸菌菌液、菌体及其代谢产物进行体外抑菌试验。结果显示,3株乳酸菌的菌液及其代谢产物具有较强的抑菌活性,而菌体没有抑菌活性。代谢产物热稳定性较好,但受pH影响较大。随着pH升高,抑菌活性逐渐降低,在弱酸性、中性和碱性条件下没有抑菌活性。代谢产物的抑菌活性不受过氧化氢酶、胃蛋白酶、胰蛋白酶的影响。     

病原菌致病性胞外产物(ECP)的研究进展

许多病原菌能分泌多种胞外产物,如蛋白酶、磷脂酶、脂肪酶、明胶酶及溶血素等。微生物分泌的明胶酶、蛋白酶、淀粉酶、磷脂酶属于胞外酶,在细菌致病中可以和溶血毒素一起进行协同作用,分解破坏宿主组织成分。笔者综述了病原菌的致病性胞外产物,以及研究胞外产物的方法。     

三株乳酸杆菌体外抑菌试验的研究

采用牛津杯法对3株乳酸杆菌菌液、菌体及其代谢产物进行体外抑菌试验,结果显示乳酸杆菌菌液及其代谢产物都有较强的抑菌活性,而菌体没有抑菌活性;代谢产物热稳定性较好,甚至能耐受100℃的高温,但受pH值的影响较大,随着pH值的升高抑菌活性逐渐降低,在弱酸、中性和碱性条件下将失去抑菌活性;代谢产物的抑菌活性不受过氧化氢酶、蛋白酶K、胃蛋白酶和胰蛋白酶等的影响。     

疯草内生真菌Undifilum oxytropis次级代谢产物分析

为阐明疯草内生真菌Undifilum oxytropis次级代谢产物的化学组成,为其生物活性研究和开发利用提供依据,以本实验室保存的疯草内生真菌Undifilum oxytropis为研究菌株,采用察氏液体培养基对该菌发酵培养30d,收集菌丝体和发酵液,按照植物化学成分传统预试方法进行预试。在此基础上用不同极性有机溶剂梯度萃取得到菌丝体和发酵液石油醚相、氯仿相、乙酸乙酯相和正丁醇相,经薄层色谱检测及气相检测方法对其次级代谢产物进行初步分析。系统预试表明,Undifilum oxytropis发酵产物中含有生物碱、蛋白质、酚类与鞣质、黄酮类、醌类、挥发油和油脂等物质;薄层色谱检测发现,Undifilum oxytropis发酵产物中至少含有5种化合物,且菌丝体和发酵液正丁醇相均有苦马豆素及其类似物存在;气相色谱分析Undifilum oxytropis发酵液中苦马豆素含量为5.17×10~(-3)g/L。疯草内生真菌Undifilum oxytropis发酵产物含有苦马豆素等多种次级代谢产物。     

PCR检测犬腺病毒方法的建立

利用人工合成的两条引物,对病犬肝脏和细胞培养物冻融上清中的1、2型犬腺病毒DNA进行多聚酶链式反应（PCR）检测;再将扩增产物进行同位素标记,与纯化病毒DNA进行打点杂交。扩增产物经电泳分析结果表明,其大小与设计的引物间的序列大小基本一致;扩增产物经标记后只与犬腺病毒DNA发生杂交反应,表明其为犬腺病毒特异性核酸片段。     

表达H5N1亚型禽流感HA、NA、 M1基因的重组杆状病毒的鉴定

为证实插入了禽流感HA、NA、M1、M2、NP基因的重组杆状病毒的表达物为具有禽流感形态的病毒样颗粒,对表达产物进行电镜、Western Blot、ELISA检测。蔗糖密度梯度离心纯化表达产物,电镜观察,可见禽流感病毒样颗粒;以Western Blot方法用HA、NA、M1抗体检测表达产物和纯化产物,可见特异性条带;NA多抗为捕获抗体、HA单抗为检测抗体建立夹心ELISA方法,检测表达产物,结果为阳性。结果显示,插入了禽流感HA、NA、M1、M2、NP的重组杆状病毒能够表达禽流感病毒样颗粒。     

Rabbit model for estimating relative bioavailability, residues and tissue tolerance of intramuscular products: comparison of two ampicillin products

A rabbit model for simultaneous investigation of the bioavailability, tissue residues and tissue tolerance of intramuscularly administered veterinary medicines is described. The bioavailability of ampicillin from two intramuscular products, which had been found to be different in calves, were compared in a two-way crossover design. The ampicillin levels in plasma, ampicillin residues in tissues, the plasma creatine kinase activity and the tissue damage at the injection sites were studied. The absolute bioavailabilities for the products were 100% and 40%. Differences in pharmacokinetics of ampicillin between sexes were observed after intravenous and intramuscular administration. Only slight tissue damage could be detected at the injection sites after intramuscular administration of these products. The results were compared with those obtained previously in calves and were found to be similar. Further investigations with other intramuscular drug products to validate this model are under way.     

转移因子注射液中细菌内毒素检测方法研究

为检测转移因子(TF)注射液中细菌内毒素,本研究采用鲎试剂与细菌内毒素产生凝集反应的凝胶法对国内1个生物制品厂的TF注射液样品进行了检测,实验结果表明当TF注射液稀释至0.025 mg/mL时,对鲎试剂的凝集反应无干扰作用.因此,该方法适用于TF注射液中细菌内毒素检测,并可以用于该项生物制品的质量监控.     

Bioavailability in the rabbit of penicillin and dihydrostreptomycin from three commercial penicillin/aminoglycoside fixed combination products for intramuscular injection

The bioavailability of penicillin and dihydrostreptomycin from three penicillin/ aminoglycoside fixed combination products for intramuscular injection was investigated in a four-way, randomized, crossover experiment in rabbits. Attention is focused on bioequivalence based on plasma concentration vs. time profiles to study whether the rabbit is a good model to detect differences in in vivo delivery of penicillin and/or dihydrostreptomycin after intramuscular administration of different products. In all products, penicillin was present as a suspension. Although the extent of absorption of penicillin did not differ between the three products, large differences in the rates of absorption were observed. With respect to dihydrostreptomycin, no significant differences were observed between the products. The results from this study demonstrate that the rabbit is a good model to detect differences in bioavailability of suspended penicillin from penicillin/dihydrostreptomycin fixed combination products for intramuscular injection. A study with the same products is presently being carried out in calves to investigate whether bioequivalence studies in rabbits could replace studies in the target animals.     

黄芩注射液的制备工艺及质量标准的初步研究

目的:研制以黄芩和柴胡为主药的注射剂,并建立质量控制标准。方法:以传统的水提醇沉法制备该注射剂,采用薄层色谱法对黄芩甙进行鉴定,并用紫外分光光度法测定黄芩甙的含量。结果:采用水提醇沉法制备得到的该注射剂符合药典关于注射剂的各项检查。实验结果表明:黄芩苷含量在1.6μg/mL～8.0μg/mL与吸光度成良好的线性关系,相关系数0.9996,平均回收率99.4%,相对标准偏差为0.6%(n=5)。黄芩甙的薄层色谱鉴别,薄层色谱斑点清晰、空白无干扰,重现性好。结论:该制剂生产工艺简单,质量标准简单易行,产品质量容易控制。     

影响注射剂澄明度因素的探讨

注射剂的澄明度是兽药生产中质量控制的主要指标,澄明度不合格的注射剂产品,不但影响药物疗效,伤害动物身体;也加大了企业生产的成本,影响了经济效益。从原辅材料、包装材料、净化系统、管道系统等方面对澄明度的影响因素进行了分析探讨,以寻求改进、提高产品质量的方法。     

Use of deep nasopharyngeal swabs as a predictive diagnostic method for natural respiratory infections in calves

An epidural puncture was performed using the lumbosacral approach in 18 dogs, and the lack of resistance to an injection of saline was used to determine that the needle was positioned correctly. The dogs' arterial blood pressure and epidural pressure were recorded. They were randomly assigned to two groups: in one group an injection of a mixture of local anaesthetic agents was made slowly over 90 seconds and in the other it was made over 30 seconds. After 10 minutes contrast radiography was used to confirm the correct placement of the needle. The mean (sd) initial pressure in the epidural space was 0.1 (0.7) kPa. After the injection the mean maximum epidural pressure in the group injected slowly was 5.5 (2.1) kPa and in the group injected more quickly it was 6.0 (1.9) kPa. At the end of the period of measurement, the epidural pressure in the slow group was 0.8 (0.5) kPa and in the rapid group it was 0.7 (0.5) kPa. Waves synchronous with the arterial pulse wave were observed in 15 of the dogs before the epidural injection, and in all the dogs after the epidural injection.     

关于进口动物产品检疫证书确认标准的探讨

作者根据《中华人民共和国进出境动植物检疫法》第十一、十二条对输出国官方出具检疫证书的规定,通过对文锦渡口岸进口动物产品随附检疫证书进行收集、分析、探讨,提出输出国官方检疫证书的确认制度和官方有效检疫证书的确认标准。并由此引发到进口动物产品检疫管理的探讨,提出加强进口动物产品检疫的五方面工作。     

A Comparison of Serum Concentrations of Penicillin After Intramuscular Injection and Subcutaneous and Deep Injection into the Dewlap in Cattle

Using an aqueous solution of sodium benzyl penicillin as the model substance, a comparison was made in cattle between absorption after intramuscular injection, and after subcutaneous and deep injection into the dewlap.The duration of supposed therapeutically effective serum concentrations using the 2 dewlap routes was longer than for the intramuscular route, although maximum concentrations were lower. The applicability of injection into the dewlap, especially the subcutaneous route, is discussed in relation to intramuscular injection.     

Expressions of protein oxidation markers, dityrosine and advanced oxidation protein products in Cisplatin-induced nephrotoxicity in rats

The purpose of this study was to evaluate whether dityrosine and advanced oxidation protein products (AOPP) reflect the severity of cisplatin-induced nephrotoxicity. Immunoexpression of dityrosine in kidneys and plasma AOPP concentration were examined up to day 4 post-cisplatin injection in rats. Cisplatin injection induced tubular injury on days 2-4 after injection and increased serum creatinine and BUN on days 3 and 4. On days 2-4, dityrosine was immunostained in the cytoplasm of damaged tubular cells, and their immunostaining intensity increased time-dependently. Plasma AOPP levels were significantly increased on days 3 and 4. These results suggest that expressions of dityrosine and AOPP were associated with the severity of renal injury and may be useful markers for the development of cisplatin-induced nephrotoxicity.     

Bioequivalence study in calves of three commercial penicillin/ dihydrostreptomycin fixed combination products for intramuscular injection

A bioequivalence study with three penicillin/dihydrostreptomycin fixed combination products for intramuscular administration was performed in dairy calves. In addition to plasma concentrations of penicillin and dihydrostreptomycin, creatine phosphokinase concentrations were determined during a period of 72 h after administration of the drug products. Considerable differences were observed in the pharmacokinetics of penicillin from the three products. Although the extent of absorption was similar for all products, one product showed a significantly slower release from the site of injection. Except for the AUC , the 90% confidence intervals for these parameters exceeded the acceptable range of 0.80-1.20. Therefore, these products are not bioequivalent with respect to the rate of absorption of penicillin. Concerning the pharmacokinetics of dihydrostreptomycin in calves, it could not be concluded that the products were bioequivalent, since the 90% confidence intervals of the ratios for C _max, t _max and MRT exceeded the range of 0.80-1.20. From this study in calves, it was also found that the product with the slowest release of penicillin from the injection site caused the most severe tissue damage, based on plasma creatine phosphokinase concentrations. Comparing the results from this study in calves with those from a previous study in rabbits, it can be concluded that the rabbit is a good animal model that could substitute for large animals, at least calves, in bioequivalence studies for penicillin/dihydrostreptomycin fixed combination products.