论“中国医院药学”

论“中国医院药学”汤光尹武华ＨｏｓｐｉｔａｌＰｈａｒｍａｃｙ这个词可以指一个工作场所，即医院药房；也可以指医院药学。从历史上来看，调配技术是药学的主要内容；随着工业和科学的发展，逐渐发展成一个药学分支学科。在我国，医院药学经历了从简单的处方调配技术发...     

我国医院药学现状与展望

医院药学是药学在医院领域中的应用 ,是研究医院的药品供应、药学技术、药事管理和临床用药的一门科学 ,是以药剂学为中心展开的药事管理和药学技术工作 ,以临床医师和病人为服务对象 ,以供应药物和指导、参与临床安全、合理、有效的药物治疗为职责 ;以治疗效果为质量标准 ,在医院特定环境下的药学科学工作。国外医院药学的发展在历史上经历了 3个时期 ,即以调配为主的传统时期 ;以药学服务为主的临床药学时期和以改善病人生活质量为目标的ＰＣ (Ｐｈａｒｍａｃｅｕｔｉｃａｌｃａｒｅ)时期[1] 。我国医院药学是在药剂科的药品供应、调剂、…     

新医改环境下药师职责的转变

随着医药卫生体制改革的不断深入，医院药学的传统观念正逐渐向以患者为中心的临床药学、药学服务等药学技术服务型转变，这就赋予了药帅提供药学服务的重要职责。本文针对在新医改环境下如何更好地履行药师职责进行了初步探讨。     

寡核苷酸配基在临床诊断和疾病治疗中的应用

利用指数级富集的配基进化(SELEX)技术筛选得到的寡核苷酸配基与靶分子的结合具有亲和力高、特异性强的特点，在临床诊断和疾病治疗中具有一定的优势，有可能发展成为一类新型药物和诊断试剂。本文从配基的筛选和改造，配基作为蛋白功能阻断剂、靶向剂和体外诊断试剂的应用和配基在应用中可能存在的问题等几个方面，对用SELEX技术筛选的寡核苷酸配基在临床诊断和疾病治疗中的应用进行综述。     

基于质量功能展开技术与卡诺模型优化慢病药学服务模式及效果评价

目的 应用质量功能展开（QFD）技术与卡诺（KANO）模型结合的方法,即QFD-KANO法,优化慢病药学服务模式,提高药学服务质量。方法 应用QFD-KANO法分析慢病诊疗过程中医师、护理人员、药师、患者的需求,针对核心需求对现有的慢病药学服务模式进行优化,比较优化前后药学服务效果。结果 慢病药学服务模式优化后,处方的平均前置拦截审核时间由10.0min降至1.5min,药品使用不合理率从0.34%降低到0.19%,每季度接受用药教育的患者例数从768例增加至每季度2301例,每季度血药浓度监测解读例数从135例提高至215例,医护技三位一体药学服务覆盖率从10.00%提升至17.00%,慢病患者满意率从92.44%提升至97.89%。结论 QFD-KANO法有助于挖掘慢病药学服务中医师、护理人员、药师和患者的核心需求,能够优化慢病药学服务模式,提升药学服务质量。     

SSCP在寡核苷酸适配子ssDNA多克隆筛选中的应用

目的利用单链构象多态性技术弥补SELEX(systematic evolution of ligands by exponential en-richment,SELEX)实验在寡核苷酸适配子克隆、测序环节中的不足,减少SELEX实验的环节。方法利用单链构象多态性技术(singel-strand conformation polymorphism,SSCP)将多轮SELEX筛选的198个寡核苷酸适配子克隆在测序前进行SSCP筛查。结果对198个克隆的SSCP电泳条带位置分析,挑选出58个克隆测序。SSCP筛查得到有效克隆的检出率为20.69%,是不用SSCP筛查检出率6.07%的3.4倍;如果去除组间误差,有效克隆的检出率44.44%是不用SSCP筛查检出率的7.32倍。结论通过对198个克隆的20批次SSCP电泳筛查,并经58个克隆测序后的实验结果证明:同一SSCP电泳胶片上电泳条带泳动位置相同的克隆片段,基因序列相同;电泳条带泳动位置不同的片段,基因序列不同。根据SSCP电泳条带泳动位置,挑选有代表性的克隆测序,避免对多克隆盲目测序,为多克隆测序前的筛查探索一条新的途径。     

现代医院临床药学咨询服务模式探讨

目的探索现代医院临床药学咨询服务模式.方法依据国家颁发的各项技术要求，针对临床出现的不合理用药现象进行研究。结果与结论设立临床药学工作机构，制定药学咨询服务措施，建立临床药学咨询服务体系的现代医院，临床药学咨询服务模式。     

南昌大学第二附属医院药学部简介

药学部在1924年医院成立时即设有,经过近百年的建设和发展,已成为集药品供应与调剂、临床药学服务与管理、药学实践与教育、药学科研与教学为一体的综合性技术服务部门。药学部下设供应保障科和临床药学科两个科室。由150余名药学专业技术人员组成,其中博士7人、硕士21人,硕士生导师2人,主任/副主任药师19人,主管药师53人。近年来,我部主持国家自然基金4项,中国博士后科学基金2项,中华医学会临床药学分会吴阶平基金项目1项,省主要学科学术和技术带头人培养计划一青年人才项目1项,省厅级项目20项。     

中国医院药学的回顾与展望

医院药学(Hospital Pharmacy)这个词可以指一个工作场所,即医院药学部门(Hospital Pharmacy Department);也可以指药学的一部分。近50年来,中国的医院药学从简单的调剂配方技术发展到一个综合性的     

Cell SELEX技术筛选巨噬细胞源性泡沫细胞适配子的初步研究

目的利用Cell SELEX技术筛选巨噬细胞源性泡沫细胞适配子。方法通过80mg/L氧化性低密度脂蛋白诱导THP-1巨噬细胞为泡沫细胞;聚合酶链反应扩增文库;生物素-链霉亲和素磁珠分离法构建单链DNA文库;以巨噬细胞源性泡沫细胞为靶细胞,巨噬细胞和血管平滑肌细胞为反筛细胞,利用Cell SELEX技术筛选与巨噬细胞源性泡沫细胞结合的寡核苷酸序列,并对最后一轮筛选结果进行克隆测序。结果 PCR的退火温度对PCR产物的量有显著的影响,退火温度由65℃提高为72℃时的产物量明显增加。利用生物素-链霉亲和素磁珠分离法获得了纯度高、产量大的ssDNA文库;经过18轮筛选后,成功获得了28个与预期大小相同的ssDNA序列。结论利用Cell SELEX技术筛选出了巨噬细胞源性泡沫细胞的适配子,为研发用于动脉粥样硬化的早期无创诊断试剂和靶向治疗药物奠定了基础。     

Low efficient generation of ssDNA aptamer using chemical modified spacer primer

Aptamers are oligonucleotides (ssDNA or RNA) with an appropriate size of 100 bps that bind with high affinity and specificity to a wide range of target molecules, including virtually any class of protein, drugs or small organic/inorganic molecules. The in vitro selection process referred to as SELEX provides a powerful tool to identify specific aptamers with high affinity and even discriminate between closely related targets. Aptamers have various applications such as analytical tools, disease diagnosis and prediction, pharmaceutical research, drug development, therapy and even for environmental monitoring. Nowadays, with the development of SELEX methods, generation of aptamer becomes more efficient, less time consuming and even automatically. The whole SELEX process includes binding, separation, and nucleic acid amplification. As amplification of nucleotides is an important process in successive SELEX, we will compare several methods for generation of aptamer in this report.     

Low efficient immobilization of active chloramphenicol for SELEX

Chloramphenicol is an antibacterial antibiotic which interferes with the protein synthesis of microorganisms. However, the use of chloramphenicol should be limited in humans and food products, because it is known to have side effects such as genotoxicity and aplastic anemia in humans. Therefore, it is important to monitor the amount of chloramphenicol in food products. Instead of using conventional analytical methods or antibodies, using aptamers can be a good alternative for measuring the amount of chloramphenicol in food. Aptamers are nucleic acids within 100 base pairs that can bind to target with high specificities and sensitivities, and are derived through a process called SELEX. Here, we report that the immobilization efficiency of chloramphenicol is low with SELEX, which can lead to the low amount of active chloramphenicol immobilized to epoxy resin. This result may be applicable to general SELEX process, especially for negative SELEX during which aptamers that are not bound to targets are removed to increase the amount of aptamers that can bind specifically to targets.     

Cell-based aptamer selection for diagnosing cancer and predicting cancer progression

Antibodies are excellent molecular recognition agents for a wide range of applications therefore they have been used heavily in clinical assays such as disease diagnosis. More recently, aptamers have emerged as alternative capturing agents in a variety of applications including medical diagnosis, environmental toxicity detection, targeted drug delivery and viral therapeutics. Aptamers are ssDNA or RNA that form three dimensional structures and bind to the target molecules such as peptide, protein or small molecules. Aptamers are generated by in vitro process called “SELEX (Systematic Evolution of Ligands by Exponential Enrichment)”. Conventionally, SELEX is performed with immobilized target molecules such as proteins in column, filter or beads. However, for some targets like membrane proteins, it is very difficult or almost impossible to immobilize the target proteins in their active conformation. However, cell-based aptamer selection technology explain how it can be better than standard immobilization methods in brief. Here, we described the cell-specific aptamers selecting technology, called cell-based SELEX, for diagnosing disease and predicting disease progression, especially in the case of complex disease, like cancer.     

Aptamers: new arrows to target dendritic cells

Aptamers, as a novel class of molecular probes for diagnosis, imaging and targeting therapy, have attracted increasing attention in recent years. Aptamers are generated from libraries of single-stranded nucleic acids against different molecules via the “systematic evolution of ligands by exponential enrichment” (SELEX) method. SELEX is a repetitive process of a sequential selection procedure in which a DNA or RNA library pool is incubated separately with target and control molecules to select specific oligonucleotide aptamers with high affinities and specificities. Cell-SELEX is a modified version of the SELEX process in which whole living cells are used as targets for the aptamers. Dendritic cell (DC) targeting, as a new therapeutic approach, can improve the efficiency of immunotherapy in the treatment of allergies and cancers. DCs use various receptors to continuously induce adaptive immunity via capture and presentation of antigens to naïve T cells. DCs are considered as the best targets in modulating immune responses against cancer, autoimmunity, allergy and transplantation. Aptamers, as a new agent, can be applied in DC targeting. The purpose of this review is to present some general concepts of aptamer production and DC targeting by aptamer molecules.     

Derivation of RNA aptamer inhibitors of human complement C5.

Specific aptamer inhibitors of the human complement C5 component were produced by the SELEX methodology of directed evolution of nucleic acid ligands. The SELEX procedure started with a pool of random-sequence, 2'F-pyrimidine-modified nuclease-stabilized RNA, and after twelve rounds of iterative C5 binding and nucleic acid amplification an evolved RNA pool was obtained which contained the highest affinity binders to the C5 protein. The evolved RNA pool was then cloned and sequenced, and individual clones were analyzed for binding and function. Twenty-eight clones (out of sixty) were identified which bound C5 (termed aptamers). Seven of these aptamers formed a closely related sequence homology family; these aptamers bound C5 with a Kd 20-40 nM and also inhibited human serum hemolytic activity. In addition, these aptamers inhibited zymosan-induced generation of C5a. Aptamer inhibition of both C5b and C5a suggests that aptamer binding inhibits cleavage of C5 by the C5 convertase of both pathways. One of the inhibitory aptamer sequences was truncated to yield a 38-mer 2'F RNA aptamer which retained C5 binding and inhibitory activity. The structure of this aptamer is predicted to be a stem-loop containing thirteen base pairs, and also containing two bulges. The affinity of this aptamer was improved by performing a second biased SELEX experiment, where the randomized starting RNA pool uses a template where the individual base compositions are biased toward a specific sequence. This second SELEX experiment produced an aptamer with a Kd of 2-5 nM which retained functional activity. Another SELEX to rat C5 produced an aptamer with binding and inhibitory properties virtually identical with the human aptamer. The human and rat aptamers are being evaluated for complement inhibition in vitro and in vivo as potential therapeutics for treatment of human disease.     

Recent developments in protein and cell-targeted aptamer selection and applications

Because of their easily modified chemical structures and wide range of targets, aptamers are ideal candidates for various applications, such as biomarker discovery, target diagnosis, molecular imaging, and drug delivery. Aptamers are oligonucleotide sequences that can bind to their targets specifically via unique three dimensional (3-D) structures. Usually, aptamers are obtained from repeated rounds of in vitro or in vivo selection termed SELEX (Systematic Evolution of Ligands by EXponential enrichment), which can generate aptamers with high affinity and specificity for many kinds of targets, such as biomedically important proteins and even cancer cells. In this review, some basic principles and recent developments in the design of SELEX process are discussed, hopefully to provide some guidelines towards performing more efficient aptamer isolation procedures. Moreover, the biomedical and bioanalytical applications of aptamers are further reviewed, based on some smart biochemical modifications of these oligonucleotide structures.     

Advances and perspectives in cell-specific aptamers

Nucleic acid aptamers, generated by an in vitro selection procedure named SELEX (Systematic Evolution of Ligands by Exponential enrichment), have been popularly used in various biomedical implementations so far. An adaptation of the conventional SELEX practice to whole living cells was referred to as cell-SELEX, which is very promising in ample sophisticated analytical, therapeutic and diagnostic applications. The current review article would like to provide a survey of contemporary developments in cell-specific aptamers, including methodology, applications and optimization of cell-SELEX.     

基于SELEX技术的疾病标志物发现新策略研究进展

生物标志物(Biomarker)是指可以标记生物体结构或功能改变的生化指标.在疾病研究中,生物标志物可供客观测定和评价机体所处的生理或病理状态,为疾病的诊断、鉴定提供辅助手段.指数富集的配体系统进化(Systematic evolution of ligands by exponential enrichment,SE...     

Nucleic Acid Aptamers: New Methods for Selection,Stabilization, and Application in Biomedical Science

The adoption of oligonucleotide aptamer is well on the rise, serving an ever increasing demand for versatility in biomedical field. Through the SELEX (Systematic Evolution of Ligands by EXponential enrichment), aptamer that can bind to specific target with high affinity and specificity can be obtained. Aptamers are single-stranded nucleic acid molecules that can fold into complex threedimensional structures, forming binding pockets and clefts for the specific recognition and tight binding of any given molecular target. Recently, aptamers have attracted much attention because they not only have all of the advantages of antibodies, but also have unique merits such as thermal stability, ease of synthesis, reversibility, and little immunogenicity. The advent of novel technologies is revolutionizing aptamer applications. Aptamers can be easily modified by various chemical reactions to introduce functional groups and/or nucleotide extensions. They can also be conjugated to therapeutic molecules such as drugs, drug containing carriers, toxins, or photosensitizers. Here, we discuss new SELEX strategies and stabilization methods as well as applications in drug delivery and molecular imaging.     

Improving the stability of aptamers by chemical modification

Ever since the invention of SELEX (systematic evolution of ligands by exponential enrichment), there has been rapid development for aptamers over the last two decades, making them a promising approach in therapeutic applications as either drug candidates or diagnostic tools. For therapeutic purposes, a durable performance of aptamers in biofluids is required, which is, however, hampered by the lack of stability of most aptamers. Not only are the nucleic acid aptamers susceptible to nucleases, the peptide aptamers are also subjective to degradation by proteases. With the advancement of chemical biology, numerous attempts have been made to overcome this obstacle, many resulting in significant improvements in stability. In this review, chemical modifications to increase the stability of three main types of aptamers, DNA, RNA and peptide are comprehensively summarized. For nucleic acid aptamers, development of modified SELEX coupled with mutated polymerase is discussed, which is adaptive to a number of modifications in aptamers and in a large extent facilitates the research of aptamer-modifications. For peptide aptamers, approaches in molecular biology with introduction of stabilizing protein as well as the switch of scaffold protein are included, which may represent a future direction of chemical conjugations to aptamers.