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。     

一例湖羊纤维素性肺炎的病理组织学观察

为探究四川省某养殖场湖羊患病情况,以1只发病湖羊为例,研究观察病羊临床症状、解剖变化,采用H.E.染色方法分析病羊实质器官病理组织学变化,并从病羊中分离出菌株Z-1,采用生理生化、16S rDNA基因测序对其鉴定,采用腹腔注射对菌株Z-1进行动物回感试验,纸片扩散法对菌株Z-1进行药敏分析。结果表明:患病羊病症有高热、剧烈咳嗽并伴有喘气,口鼻周围可见黄色浓稠分泌物;解剖变化为多个实质器官明显肿大、充血,肺脏充血、颜色暗红,切开有血样泡沫状液体流出;病理组织切片可见肺、肝、脾等组织细胞肿胀、充血、出血,肺泡腔和细支气管中有大量粉红色纤维素性渗出物,炎性细胞浸润;菌株Z-1为革兰氏阳性菌,乳糖、七叶苷和山梨醇等生化反应呈阳性反应,木糖醇、甘露糖和蜜二糖等生化反应呈阴性反应;16S rDNA基因目的片段长度为1 428bp,与菌株NR_026471的相似性达到98%,综合鉴定为链球菌;动物回感试验表明该菌株对小鼠具有很强的致病性;药敏试验表明该菌株对氧氟沙星、诺氟沙星、环丙沙星和庆大霉素等敏感,对氨苄西林、青霉素和拉氧头孢等耐药。综上,该病湖羊患链球菌感染的纤维素性肺炎。     

乌鲁木齐市冬春两季宠物源葡萄球菌的耐药性及耐药基因比较

采用琼脂稀释法,检测从乌鲁木齐市部分宠物医院冬季和春季分离鉴定的154株宠物源葡萄球菌(冬季75株、春季79株)对临床常用的12种抗菌药物(阿米卡星、克林霉素、硫酸庆大霉素、氟苯尼考、四环素、利福平、恩诺沙星、青霉素、苯唑西林、阿莫西林/克拉维酸、头孢噻呋、左氧氟沙星)的耐药性;通过卡方检验,比较供试菌耐药性差异;采用PCR方法,检测葡萄球菌的5类9种耐药基因。对抗菌药物的耐药结果显示:冬季葡萄球菌对苯唑西林和青霉素的耐药率高于90.0%,对头孢噻呋和四环素的耐药率高于50.0%;春季葡萄球菌对左氧氟沙星耐药率达92.4%,对苯唑西林和氟苯尼考耐药率高于80.0%。卡方检验结果显示:冬季葡萄球菌仅对苯唑西林的耐药率显著高于春季葡萄球菌(P0.05);春季葡萄球菌对氟苯尼考、利福平、左氧氟沙星、恩诺沙星和阿莫西林/克拉维酸的耐药率均显著高于冬季葡萄球菌(P0.05)。多药耐药差异性结果显示:冬季葡萄球菌多药耐药集中在1～3耐;春季葡萄球菌多药耐药集中在4、6、9耐。基因检测结果显示:冬季葡萄球菌主要检出的基因有ermB(40.0%,30/75)和fexA(22.7%,17/75);春季葡萄球菌主要检出的基因有fexA(39.3%,31/79)和femA(20.3%,16/79),冬、春季葡萄球菌均未检出cfr、fexB和tetA基因。     

沉淀法制备的紫杉醇白蛋白微粒包封率测定

采用沉淀法制备紫杉醇白蛋白微粒,采用高效液相色谱法(HPLC)测定紫杉醇质量浓度,以二氯甲烷为有机相,采用萃取法分离白蛋白微粒与游离药物,测定其包封率。结果表明:紫杉醇在10~100 mg·L-1范围内线性良好,平均回收率可达到97.6%~101.0%;有机溶剂萃取法测定紫杉醇白蛋白微粒包封率的平均回收率,达94.80%~101.67%。这种方法测定紫杉醇白蛋白微粒回收率准确可靠,适用于沉淀法制备的白蛋白微粒的包封率测定。     

基于最新诊疗方案中抗新型冠状病毒药物的回顾分析与药学监护

2019年12月以来,湖北武汉爆发由新型冠状病毒引发的病毒性肺炎疫情并迅速蔓延,现已确认该新型病毒为SARS-CoV-2.由于目前针对该病毒尚无确切有效的抗病毒药物,给我国公共卫生事业带来了严峻的考验.随着疫情的发展,国家卫生健康委员会下发了最新的《新型冠状病毒肺炎诊疗的方案(试行第七版)》.该文从临床药学角度对最新诊疗方案中所涉及的抗病毒药物进行回顾分析,提出药学监护的重点,并对潜在有效的药物进行展望,以期为抗击新型冠状病毒疫情提供药物使用参考.     

A New Bioassay Platform Design for the Discovery of Small Molecules with Anticancer Immunotherapeutic Activity

Immunotherapy takes advantage of the immune system to prevent, control, and eliminate neoplastic cells. The research in the field has already led to major breakthroughs to treat cancer. In this work, we describe a platform that integrates in vitro bioassays to test the immune response and direct antitumor effects for the preclinical discovery of anticancer candidates. The platform relies on the use of dendritic cells that are professional antigen-presenting cells (APC) able to activate T cells and trigger a primary adaptive immune response. The experimental procedure is based on two phenotypic assays for the selection of chemical leads by both a panel of nine tumor cell lines and growth factor-dependent immature mouse dendritic cells (D1). The positive hits are then validated by a secondary test on human monocyte-derived dendritic cells (MoDCs). The aim of this approach is the selection of potential immunotherapeutic small molecules from natural extracts or chemical libraries.     

肯塔基沙门菌对氟喹诺酮和碳青霉烯类药物耐药性研究进展

肯塔基沙门菌（Salmonella Kentucky）是继鼠伤寒沙门菌和肠炎沙门菌后又一与人类肠道疾病相关的沙门菌血清型，因其获得和传播耐药性的能力较强而逐渐引起关注。该血清型沙门菌按照多位点序列分型分得的众多亚型中，ST198血清亚型具有易获得耐药突变的基因组岛的特性，21世纪以来不断进化，除获得常见抗菌药物耐药基因外，还相继获得对氟喹诺酮类和碳青霉烯类药物的耐药性并扩散全球，造成临床上难以治疗的病例而引起世界卫生组织的关注。作者整理了肯塔基沙门菌的进化历程，着重阐述肯塔基沙门菌关于氟喹诺酮类及碳青霉烯类药物的耐药情况及耐药机制，以期为中国沙门菌的实验室防控监测系统提供理论依据。     

Marine Anticancer Agents: An Overview with a Particular Focus on Their Chemical Classes

The marine environment is a rich source of biologically active molecules for the treatment of human diseases, especially cancer. The adaptation to unique environmental conditions led marine organisms to evolve different pathways than their terrestrial counterparts, thus producing unique chemicals with a broad diversity and complexity. So far, more than 36,000 compounds have been isolated from marine micro- and macro-organisms including but not limited to fungi, bacteria, microalgae, macroalgae, sponges, corals, mollusks and tunicates, with hundreds of new marine natural products (MNPs) being discovered every year. Marine-based pharmaceuticals have started to impact modern pharmacology and different anti-cancer drugs derived from marine compounds have been approved for clinical use, such as: cytarabine, vidarabine, nelarabine (prodrug of ara-G), fludarabine phosphate (pro-drug of ara-A), trabectedin, eribulin mesylate, brentuximab vedotin, polatuzumab vedotin, enfortumab vedotin, belantamab mafodotin, plitidepsin, and lurbinectedin. This review focuses on the bioactive molecules derived from the marine environment with anticancer activity, discussing their families, origin, structural features and therapeutic use.     

微流控芯片在农产品安全检测中的应用

近年来,随着食品工业的迅猛发展,国内外食品安全事故也频繁发生。开发安全可靠、便携有效的检测方法对食品安全至关重要。微流控芯片集样品前处理、分离和检测等过程为一体,实现了样品前处理和检测分析的自动化、集成化、小型化、低消耗、高效率和便携化,满足了现场、实时检测的需求。本研究对微流控芯片在食品成分、农药残留、兽药残留、食品添加剂和重金属检测中的应用进行归纳总结,为农产品安全检测提供参考。     

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.