甘露糖修饰的壳聚糖聚乳酸-羟基乙酸共聚物纳米微球作为口蹄疫病毒核酸疫苗递送载体的特性评价

旨在对制备的甘露糖修饰的壳聚糖聚乳酸-羟基乙酸共聚物[poly （D，L-lactide-co-glycolide），PLGA]纳米微球作为口蹄疫病毒（foot-and-mouth disease virus，FMDV）核酸疫苗递送载体进行评价。采用西佛碱反应和元素分析制备具有一定取代度的甘露糖修饰的壳聚糖衍生物（mannose modified chitosan，MCS），然后，经双重乳化挥发法制备得到甘露糖修饰的壳聚糖PLGA纳米微球（MCS-PLGA-NPs）。采用纳米粒径仪检测MCS-PLGA-NPs粒径分布和表面电势（zeta）、扫描电镜考察其形态、琼脂糖凝胶电泳观察其对质粒的吸附和吸附质粒后抵抗核酸酶降解能力、CCK-8法检测MCS-PLGA-NPs的细胞毒性、激光共聚焦观察巨噬细胞对MCS-PLGA-NPs-质粒DNA复合物的摄取、荧光显微镜和Western blot验证MCS-PLGA-NPs加载质粒DNA在细胞中的表达。元素分析结果表明，成功制备了取代度为5%~10%的MCS。纳米粒径测定和扫描电镜结果表明，MCS-PLGA-NPs的zeta为正值、粒径分布均匀且形态规则呈球形。琼脂糖凝胶电泳结果显示，MCS-PLGA-NPs吸附质粒的能力随着其质量的增加而增强并且可以在一定程度上抵抗核酸酶降解质粒DNA。在细胞毒性试验中，不同浓度的MCS-PLGA-NPs与RAW264.7细胞共孵育24 h后，细胞存活率仍在85%以上。在细胞摄取试验中，用激光共聚焦显微镜可以明显观察到质粒DNA结合到纳米微球表面被RAW264.7细胞摄取。荧光显微镜和Western blot试验证明MCS-PLGA-NPs加载质粒DNA可以在细胞中进行表达。综上表明，本研究成功制备了MCS以及具有递送核酸疫苗能力的MCS-PLGA-NPs，为FMDV核酸疫苗的递送研究提供了新的方向和见解，也为该递送载体携带特定抗原靶向抗原递呈细胞表面甘露糖受体以及应用于动物免疫的研究奠定基础。     

Application of imidacloprid controlled-release granules to enhance the utilization rate and control wheat aphid on winter wheat

During winter wheat production, aphids need to be controlled with pesticides for the entire growth period. Controlled-release technology has been regarded as an alternative method for the improvement of pesticide efficiency. This study investigated two types of imidacloprid controlled-release granule (CR-GR): 2% imidacloprid CR-GR and 0.2% imidacloprid pesticide-fertilizer controlled-release granule (PF-CR-GR) when wheat was sown in winter. The release performance, utilization rate, terminal residues in edible parts, control effect on aphids, and achieved winter wheat yield were evaluated for both laboratory experiments and field application. Imidacloprid PF-CR-GR released more quickly in aqueous medium than CR-GR because of its good water solubility. After CR-GR treatments, the concentrations in wheat roots and soil were similar throughout the entire sampling period, and the concentrations in shoots were about 10–20% of those in roots. Imidacloprid was better absorbed when CR-GR was used as root treatment, compared with foliar treatment. Field application showed that imidacloprid CR-GR and PF-CR-GR controlled aphids throughout the entire growth period of winter wheat and improved the wheat yield. These findings identified application of imidacloprid CR-GR and PF-CR-GR on winter wheat as an effective way to enhance the pesticide utilization rate and ensure adequate yield. This paper provides a theoretical basis for the scientific use of pesticides and guides scientific pesticide application.     

Preparation of Alginate-Based Biomaterials and Their Applications in Biomedicine

Alginates are naturally occurring polysaccharides extracted from brown marine algae and bacteria. Being biocompatible, biodegradable, non-toxic and easy to gel, alginates can be processed into various forms, such as hydrogels, microspheres, fibers and sponges, and have been widely applied in biomedical field. The present review provides an overview of the properties and processing methods of alginates, as well as their applications in wound healing, tissue repair and drug delivery in recent years.     

60Co-γ射线对9种庭院月季扦插苗的辐射效应研究

为探讨⁶⁰Co-γ射线对不同庭院月季品种扦插苗的辐射诱变效应,以粉扇、金凤凰、梅朗口红、紫袍玉带、安吉拉、粉色达芬奇、你的眼睛、香槟和九重香9个庭院月季品种的扦插苗为研究对象,⁶⁰Co-γ射线为辐照源,设置不同剂量(0、25、45、60、80 Gy)进行辐射处理,测定了死亡率、萌芽率、变异率和主要的形态指标。结果表明,随着辐射剂量增加,月季扦插苗平均死亡率逐渐增加,从33.61%(0 Gy)上升至79.71%(80 Gy);植株生长受到显著抑制,植株变异率呈先升高后降低的趋势,辐射剂量为45 Gy时,植株变异率达到最高,占总体变异率(4.20‰)的64.70%。此外,不同品种月季对⁶⁰Co-γ射线辐射的敏感性不同,半致死辐射剂量也存在差异,其有效值介于26~55 Gy之间。综合⁶⁰Co-γ射线对月季扦插苗死亡率、生长势和变异率的影响,推荐庭院月季扦插苗的最适辐射剂量为45 Gy,这为月季辐射育种提供了理论基础。     

Four years validation of decision support optimising herbicide dose in cereals under Spanish conditions

The Danish decision support system Crop Protection Online (CPO) optimises herbicide weed control. CPO recommends specific herbicide solutions to achieve a required level of control. The aim is to apply herbicides as little as possible but as much as necessary. CPOWeeds is a version of CPO adjusted to conditions in North-eastern Spain. The predicted efficacies and the yield obtained with CPOWeeds were validated in winter cereal field trials from 2010 to 2013. All CPOWeeds treatments were related to the efficacies obtained with standard herbicide treatments decided upon by local advisors. The predictions from CPOWeeds were compared to the actually achieved efficacies in the field trials for the nine weed species at different developmental stages and for 84.2% of the comparisons the obtained efficacies were equal to or higher than predicted. The average difference between predicted and observed efficacies was 2.35 percentage points. Yield was measured in three trials and the recommendations from CPOWeeds were maintaining yield. There were two situations where CPOWeeds were performing suboptimal. One is in the early weed growth stages, as the model is not yet prepared to account for water stress on root action herbicides applied at 10-11 BBCH. The second situation was in fields with a prior unidentified population of resistant Alopecurus myosuroides. For key species in winter cereals in Spain, such as Avena sterilis, Lolium rigidum and Papaver rhoeas, CPOWeeds achieved a satisfactory control level. It was concluded that the use of CPOWeeds allowed optimisation of the herbicide application with a very high robustness. The recommendations were satisfactorily for the conditions of the Northeast of Spain and have the potential to decrease the amount of applied herbicides by at least 30%. Therefore, it can be an important tool in Integrated Weed Management.     

禽致病性大肠杆菌luxS和rbsC双基因缺失株的构建及其生物特性分析

禽致病性大肠杆菌(APEC)能引起家禽严重的呼吸系统和全身性疾病,由于APEC耐药菌株的不断出现迫使新型抗菌方式的研发迫在眉睫。本研究旨在评价APEC的luxS和rbsC双缺失的突变株作为减毒疫苗候选株的潜力,通过在APEC分离株DE17的单基因缺失株DE17ΔluxS的基础上,利用Red重组系统构建双基因缺失株DE17ΔluxSΔrbsC,并对其生物学特性进行了分析。结果显示:rbsC的缺失不影响DE17ΔluxS的生长特性和生物被膜形成能力,但运动性显著降低;与DE17ΔluxS相比,DE17ΔluxSΔrbsC的黏附能力和入侵能力分别降至46.92%和28.78%;半数致死量(LD50)结果显示,毒力下降了115.5倍;肝脏、脾脏和血液中的载菌量分别下降了3.63倍、79.20倍和2124.41倍。根据以上结果推测,rbsC基因的缺失会进一步降低DE17ΔluxS的毒力,本文为评价DE17ΔluxSΔrbsC株减毒机制提供参考。     

两种杀虫剂亚致死剂量对龟纹瓢虫捕食功能反应的影响

为了协调化学防治和生物防治二者间的矛盾,掌握药剂对龟纹瓢虫捕食功能的影响,利用吡虫啉和高效氯氰菊酯对龟纹瓢虫的LC_(25)和对豌豆蚜的LC_(25)分别处理2种昆虫,测定了2种杀虫剂亚致死剂量对龟纹瓢虫捕食豌豆蚜功能反应的影响。结果表明,龟纹瓢虫捕食功能反应模型没有发生变化,仍是典型的Holling-Ⅱ型,但模型的各项参数发生了改变。高效氯氰菊酯直接处理龟纹瓢虫,对其捕食能力抑制最强;龟纹瓢虫捕食吡虫啉处理后的豌豆蚜,对其捕食能力抑制最弱。龟纹瓢虫以2种方式受药后,瞬时攻击率a下降,最大为0.860 4,最小为0.782 3,相对于对照组(CK)下降5.21%~15.78%;处理猎物的时间T_h延长,最长为0.017 6 d,最短为0.013 7 d,相对于CK延长16.10%~49.15%;最大理论捕食量下降,最大为73.09头,最小为56.74头,相对于CK下降14.00%~33.24%;药剂处理18h后,捕食速率均显著(P0.05)低于同时间下的对照,18h时最大为1.67头/h,最小为1.56头/h,相对于同时间时的CK降低13.04%~18.84%,24 h后最大为1.66头/h,最小为1.46头/h,为同时间下CK的9.14%~20.00%。因此,不论龟纹瓢虫以何种方式受药,都不利于其捕食豌豆蚜。其中,高效氯氰菊酯直接作用于龟纹瓢虫时负面影响最大,但不改变其对豌豆蚜的捕食功能反应模型。     

不同辐照剂量对蜂球囊菌杀灭效果的影响

本试验以蜂球囊菌(Ascosphaera apis)为研究对象,探究不同辐照剂量对蜂球囊菌的杀灭效果,以期明确有效杀灭蜂球囊菌的最低辐照剂量,为Co60γ射线辐照蜜蜂饲料(特别是蜂花粉)时的辐照剂量选择提供依据。利用细菌纯化技术从患白垩病意蜂幼虫体内分离纯化得到蜂球囊菌,并结合形态学、乳酸酚棉兰染色及5.8SrDNA序列分析技术进行鉴定;同时制备不同梯度浓度的蜂球囊菌孢子悬液加入到蜜蜂幼虫饲料中,以饲喂方式侵染3日龄意蜂幼虫,确定半数致死浓度(LC50);蜜蜂幼虫饲料中添加上述确定LC50的孢子,以不同辐照剂量处理,并通过侵染幼虫试验,确定有效杀灭蜜蜂饲料中孢子的最低辐照剂量。结果表明,通过形态学和分子生物学鉴定,从患白垩病的意蜂幼虫体内分离纯化得到真菌即为蜜蜂白垩病的病原——蜂球囊菌;蜂球囊菌对人工饲养条件下的意蜂幼虫的LC50为9.5×104个/mL;当Co60γ射线辐照剂量为7.0kGy时,幼虫患病率与未辐照组差异显著(P0.05),由此确定对添加半数致死浓度的蜂球囊菌孢子饲料的最低有效辐照剂量为7.0kGy。