Simple and Rapid Quality Control of Sulfated Glycans by a Fluorescence Sensor Assay—Exemplarily Developed for the Sulfated Polysaccharides from Red Algae Delesseria sanguinea

Sulfated polysaccharides (SP) from algae are of great interest due to their manifold biological activities. Obstacles to commercial (especially medical) application include considerable variability and complex chemical composition making the analysis and the quality control challenging. The aim of this study was to evaluate a simple microplate assay for screening the quality of SP. It is based on the fluorescence intensity (FI) increase of the sensor molecule Polymer-H by SP and was originally developed for direct quantification of SP. Exemplarily, 65 SP batches isolated from the red alga Delesseria sanguinea (D.s.-SP) and several other algae polysaccharides were investigated. Their FI increase in the Polymer-H assay was compared with other analytical parameters. By testing just one concentration of a D.s.-SP sample, quality deviations from the reference D.s.-SP and thus both batch-to-batch variability and stability can be detected. Further, structurally distinct SP showed to differ in their concentration-dependent FI profiles. By using corresponding reference compounds, the Polymer-H assay is therefore applicable as identification assay with high negative predictability. In conclusion, the Polymer-H assay showed to represent not only a simple method for quantification, but also for characterization identification and differentiation of SP of marine origin.     

基于量子点的赭曲霉毒素AsFLISA检测技术研究

建立了基于量子点标记技术的赭曲霉毒素A(Ochratoxin A,OTA)双抗夹心荧光免疫检测(sandwich fluorescence-linked immunosorbent assay,sFLISA)体系,包含多克隆包被抗体、生物素标记的多克隆检测抗体、量子点标记的链霉亲和素等,获得了sFLISA的最佳操作参数:包被抗体浓度2.5μg/mL,检测抗体稀释500倍,量子点标记链霉亲和素稀释100倍。该方法在OTA浓度3.125~125μg/L之间时,相对荧光强度和OTA浓度呈线性关系,回归方程为y=0.0206x+0.2018,R2=0.9924;加标回收率在90.1%~110.0%之间,变异系数均小于10%,能较好地进行赭曲霉毒素A的定量检测。     

菲、芘对蚕豆的氧化胁迫和DNA损伤

以菲和芘为多环芳烃(PAHs)的代表物,以超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)活性和丙二醛(MDA)含量为指标,研究了菲、芘对蚕豆的氧化胁迫;利用彗星实验分析了菲、芘对蚕豆DNA的损伤效应;将蚕豆幼苗根经抗氧化剂维生素E预处理后,暴露于菲、芘污染,研究了DNA损伤与氧化胁迫间的关系。结果表明,供试条件下菲、芘污染导致蚕豆幼苗SOD、POD、CAT活性提高和MDA含量上升,且MDA含量与菲、芘浓度均显著正相关;蚕豆根尖细胞DNA损伤随菲、芘暴露浓度的升高而增大,0~50 mg·kg-1菲污染条件下彗星图像尾矩(TM)值从46.41μm(阴性对照)增加到122.04μm(50 mg·kg-1菲污染处理),增大了162.96%。50mg·kg-1芘暴露下TM值从阴性对照的44.30μm增至110.36μm,增大了149.21%。经抗氧化剂维生素E预处理,蚕豆的DNA损伤程度减小。综上可知,菲、芘对蚕豆产生氧化胁迫并造成根尖细胞DNA损伤,菲、芘诱导的DNA损伤与氧化胁迫有关。     

Toxicological effects of TiO2 and ZnO nanoparticles in soil on earthworm Eisenia fetida

Nanoparticles (NPs) of TiO₂ and ZnO are receiving increasing attention due to their widespread applications. To evaluate their toxicities to the earthworm Eisenia fetida (Savigny, 1826) in soil, artificial soil systems containing distilled water, 0.1, 0.5, 1.0 or 5.0 g kg⁻¹ of NPs were prepared and earthworms were exposed for 7 days. Contents of Zn and Ti in earthworm, activities of antioxidant enzymes, DNA damage to earthworm, activity of cellulase and damage to mitochondria of gut cells were investigated after acute toxicity test. The results from response of the antioxidant system combined with DNA damage endpoint (comet assay) indicated that TiO₂ and ZnO NPs could induce significant damage to earthworms when doses were greater than 1.0 g kg⁻¹. We found that Ti and Zn, especially Zn, were bioaccumulated, and that mitochondria were damaged at the highest dose in soil (5.0 g kg⁻¹). The activity of cellulase was significantly inhibited when organisms were exposed to 5.0 g kg⁻¹ of ZnO NPs. Our study demonstrates that both TiO₂ and ZnO NPs exert harmful effects to E. fetida when their levels are higher than 1.0 g kg⁻¹ in soil and that toxicity of ZnO NPs was higher than TiO₂.     

酶联免疫吸附分析技术在农药残留分析中的应用

简述酶联免疫吸附分析技术的原理、方法类型以及关键的技术要点,并对该技术在农药残留分析检测中的应用前景进行了展望。     

雏鸡感染巨型艾美耳球虫后细胞免疫的研究

通过用地塞米松处理鸡和用特异性抗原刺激因子的微量全淋巴细胞转化试验（ＬＰＡ），研究雏鸡抗Ｅｉｍｅｒｉａｍａｘｉｍａ再感染中细胞免疫现象。结果表明，地塞米松处理可损伤鸡抗Ｅ．ｍａｘｉｍａ再感染的免疫力，这提示了ＣＤ８＋Ｔ细胞，γ－ＩＦＮ及ＩＬ－２等在抗．Ｅｍａｘｉｍａ再感染中的作用，利用Ｅ．ｍａｘｉｍａ卵囊抗原作刺激因子的微量全血ＬＰＡ结果表明，其刺激指数值与鸡抗Ｅ．ｍａｉｘｉｍａ再感染抵抗力有关。     

新生牛血清促细胞生长活性检测方法的建立与应用

牛血清是医药生物技术产品中重要的原辅材料之一,做好牛血清的质量控制是提高生物制品质量的重要环节。本研究通过应用PK15、BHK21和ST三种兽用生物制品研制中常用的细胞作为载体,选用国内同等价位5个不同厂家生产的新生牛血清作为比较对象,采用细胞倍增时间测定法、细胞克隆形成率测定法、细胞三次连续传代培养法、MTT比色法和微量终点稀释法五种血清质量鉴定的方法进行比较,检测新生牛血清的促细胞生长活性,最终确定了MTT比色法作为便捷高效快速筛选新生牛血清的检测方法。     

5种杀虫剂的烟青虫防治效果研究

５种杀虫剂对烟青虫的毒力试验表明：质量分数为８０％～８３％的高效氯氰菊酯和８６．７％的氰戊菊酯分别稀释２０００和１０００倍对烟青虫的防治效果较好，ＬＧ５０分别为０．０３２２ｕｇ．头＾－１和０．０５８２ｕｇ．头＾－１，久效磷和乙酰甲胺磷稀释１０００倍的效果次之，吡虫啉效果最差。     

犬瘟热病毒抗体检测方法的比较研究

用犬瘟热（ＣＤ）弱毒和自制的高免犬血清，建立了检测犬瘟热病毒（ＣＤＶ）抗体的中和（ＳＮ）试验、间接荧光抗体技术（ＩＦＡＴ）和间接免疫酶染色法。这３种方法对３８份ＣＤ弱毒疫苗免疫犬血清抗体效价的测定结果表明，当被检血清稀释度为１∶１００时，ＳＮ试验、ＩＦＡＴ和间接免疫酶染色法测定时的阳性血清数分别为２９，７和３１份，ＩＦＡＴ和间接免疫酶染色法测定结果相对于ＳＮ试验结果的符合率分别为４２．１％和９４．７％。同时发现，当ＳＮ试验测定的血清效价在１∶１００以上时，ＩＦＡＴ测定结果总是在１∶２５以上，间接免疫酶染色法测定结果总是在１∶１００以上。３种方法以各自初步确定的ＣＤＶ血清抗体效价完全保护值指标计算所测血清的完全保护率，ＳＮ试验为７６．３％，ＩＦＡＴ为７８．９％，间接免疫酶染色法为８１．６％。据此认为，ＩＦＡＴ和间接免疫酶染色法均可部分代替ＳＮ试验用于ＣＤ疫苗免疫效果的评价、免疫犬抗体水平的监测以及ＣＤ的流行病学调查     

利福平乙基纤维素微球在小鼠体内的组织分布

以有机相分散、溶媒扩散二步法制备了利福平乙基纤维素微球，粒径范围2.5-12.5μm，其中粒径2.5-5.0μm的约占63％。采用微生物测定法研究了利福平乙基纤维素微球静脉给药后，利福平在小鼠体内的组织分布，且与游离利福平在小鼠体内组织分布进行了比较，小鼠单剂量静脉注射游离利福平或利福平乙基纤维素微球（相当于利福平原药）10mg/kg，测定0.5、1、3、7、12、24、48、72、120、168h共10个时间点各组织中的药物浓度。结果表明，游离利福平给药组和利福平乙基纤维素微球给药组在12h内均以肝脏中药物浓度为最高，为9.96-22.41μg/g，在3h内微球给药组高于游离药物给药组，7、12h微球给药组的药物浓度迅速下降，低于游离药物给药组；肾脏的药物浓度在12h内微球给药组明显低于游离药物给药组，24－168h微球给药组的药物浓度下降迟缓，显著高于游离药物给药组直到168药物浓度始终是微球给药组高于游离药物给药组，12h游离药物给药组已不能检出药物含量，而微球给药组直到168h仍维持在5.44μg/g，接近游离药物给药组的最高峰值；血清中利福平浓度在48h已不能检出，而微球给药组在168h为1.33mg/L；心脏、用脏微球给药组和游离药物给药组的药物浓度变化无统计学意义，与游离药物相比，利福平微球明显提高了肺脏中的药物分布，降低了利福平对肝脏、肾脏的毒性，并显著延缓了药物代谢，说明利福平微球具有长效、靶向作用。