细胞促渗肽在透皮给药中的应用

细胞促渗肽（CPP）最早于细胞研究中发现,此后被引入透皮制剂领域,相关透皮机制尚未阐明。CPP在透皮制剂领域中应用广泛,对小分子药物、多肽、蛋白质、核酸和胶粒载体均有皮肤促渗作用。本文综述CPP在透皮领域中的应用。     

细胞内siRNA传递：应用修饰的细胞穿透肽从内涵体逃逸

RNA干扰介导的基因沉默是一种有前景的基因疗法，其中使RNA安全有效地进入胞浆很重要。细胞膜是RNA传递的首要障碍，也最难跨越，带负电性的细胞膜强烈排斥RNA。各种阳离子聚合物已被作为RNA载体，与RNA相互作用，掩盖其负电荷，形成细胞穿透性复合物。阳离子型细胞穿透肽（cell-penetrating piptide，CPP）能携RNA一起穿过细胞膜进入细胞内部，在以CPP为基础的RNA传递中，大多数RNA被细胞内吞后包埋在内涵体中。本文综述基于CPP的RNA递送方法及RNA从内涵体逃逸的策略。     

临床药学专业社会药房实习现状调查分析

目的 调查临床药学专业社会药房实习(CPP)课程的开展情况,分析该课程存在的问题,以期改进CPP课程。方法 根据已发表的CPP课程调查问卷,开展横断面研究。以中国药科大学临床药学专业本科生为调查对象,开展关于CPP课程的内容、障碍以及学生对该课程的态度、能力提升和改进建议等方面的问卷调查。结果 学生表示CPP课程帮助他们了解社会药房销售的药物,但大多数学生对CPP课程内容表示不满意,与药学监护相关的教学内容比重低（占比39.21%）,希望课程内容突出临床药学专业特色;绝大多数学生就业不考虑社会药房;大于50%的学生认为由于我国国情,社会药房不能承担药学服务相关实习任务,社会药房的执业药师不能胜任实习带教工作,需要加强执业药师的专业水平和带教能力。结论 CPP课程目前尚存在许多问题,该课程需要结合我国国情和相关政策并借鉴国外的教学经验进一步改进;加强执业药师的师资培训并建立CPP带教标准有助于推动我国高校CPP课程的发展。     

氨磷汀在血液系统疾病中的应用

细胞保护剂已成为抗肿瘤药物研究与开发领域的＂新星＂,氨磷汀就是其中的一名佼佼者,在抗肿瘤、逆转耐药和保护正常细胞方面具有潜在的应用价值,目前已被广泛用于临床多种血液系统疾病。     

蛋白质转导的研究进展

生物膜的脂质双分子层对极性强的蛋白质、DNA及高分子复合物是一个有力的屏障，使用具有临床应用价值的蛋白质等都必须克服这个障碍。研究发现，一些蛋白质上具有蛋白质转导区(PTD)的小片段，能够有效地通过生物膜，几乎进入所有细胞。以PTD为载体，通过非共价或共价结合，已成功地携带多种物质进入细胞或进入机体的组织器官。其机制可能与细胞膜上带负电荷的硫酸肝素有关。蛋白质转导在运送蛋白质、寡核苷酸或其他物质治疗病毒感染、特异性杀伤肿瘤细胞、神经退行性疾病等方面具有巨大的潜力。     

宫腔镜联合腹腔镜诊治慢性盆腔痛的临床疗效分析简

慢性盆腔痛（chronic pelvic pain,CPP）是指由各种功能性或（和）器质性原因引起的以骨盆及其周围组织疼痛为主要症状,时间超过6 个月的一组疾病或综合征[1].CPP 是妇科疾病中最常见的症状之一,据有关调查显示,40%的适龄女性患有此病,2%~5%的育龄女性生活受到CPP 的严重干扰.但CPP 有发病隐匿、病因复杂和诊治困难等临床特点,使妇科医生在诊断时受到诸多因素的干扰.微创外科的发展为CPP的诊治提出新思路,美国妇产科学会推荐对CPP 患者首选腹腔镜技术[2].而宫腔镜在诊治宫内病变上能提供有价值的建议,有助于探索宫内病变的原因.本文回顾性分析我院近年来接收的采用宫腔镜联合腹腔镜诊治的CPP 患者的临床资料,以探讨宫腔镜联合腹腔镜在慢性盆腔痛诊治中的临床应用.     

Research strategies and frontier technology advances in intracerebral pharmacokinetics北大核心CSCD

单细胞多组学测序技术是指在单个细胞水平上,对基因组学、表观基因组学、转录组学和蛋白质组学等,进行高通量测序分析的一项新技术,其能够揭示单个细胞的基因结构和基因表达状态,反映细胞间的异质性,在肿瘤学、发育生物学、免疫学及神经科学等领域发挥重要作用。随着单细胞技术的飞速发展,生命科学领域已经逐步进入了单细胞生物学时代。单细胞多组学测序数据联合分析,探索复杂的生命现象的发生和调控过程,相对于评估群体的传统分析方法,这些基于单个细胞上的分析能更加全面地描述细胞特性,可极大促进科学家们对基础科学领域的研究。单细胞技术已被广泛应用于药理学领域的研究,包括促进临床诊断与个体化治疗,以及药物开发、探究肿瘤耐药机制、解析不同疾病中的微生物菌群、优化生物医学实验模型、阐明天然产物和中药活性分子作用机制、与深度学习结合预测药物响应等。该综述重点介绍单细胞技术的最新进展,举例概述单细胞技术在药理学研究中的具体应用,并就该领域存在的挑战和未来发展趋势进行展望。     

蓖麻蛋白诱导HeLa细胞凋亡的分子机制(英文)

目的:研究蓖麻蛋白引起的Hela细胞凋亡的形态变化及机制。方法:扫描电镜,透射电镜,Western blot,细胞周期分析、细胞毒性和细胞相对存活率测定。结果:蓖麻蛋白0.05μmol·L~(-1)引起HeLa细胞发生典型的凋亡。凋亡细胞主要表现为胞浆膜起泡,核染色质浓缩,形成新月状核或膜包裹核染色质的凋亡小体;Western blot未检测到p53、Bax和ICE的p20活性亚基,而检测到CPP32的p17活性亚基,CPP32活性升高,而ICE活性无显著改变。结论:CPP32参与了蓖麻蛋白诱导的HeLa细胞凋亡过程。     

CT、眼科超声波断层诊断装置应用于眼科临床诊断的价值比较

随着现代医学科学的进展,在眼科领域也相继应用了电子计算机断层扫描装置和眼科超声波断层诊断装置(以下简称为OT和眼科超声波)于临床,提高了诊断效果。目前CT的发展已进入第六代,眼科超声波已从开始的A型进入到M型,使其扫描时间     

刺五加注射液的临床应用

刺五加注射液为五加科植物刺五加的茎叶经水醇法提取制成的棕红色灭菌溶液,含有榛皮酊、丁香苷和金丝桃苷等多种皂苷、多糖和黄酮类物质的混合物.随着刺五加注射液药理研究的深入,刺五加注射液已广泛应用于临床多种疾病的治疗,疗效确定,随着临床应用与研究领域的逐步进展,其显示出良好的应用前景,本文结合近年的文献,就其在临床多种疾病中的应用与研究综述如下.     

The cutting-edge technologies of siRNA delivery and their application in clinical trials

siRNA therapeutics allows precise regulation of disease specific gene expression to treat various diseases. Although gene silencing approaches using siRNA therapeutics shows some promising results in the treatment of gene-related diseases, the practical applications has been limited by problems such as inefficient in vivo delivery to target cells and nonspecific immune responses after systemic or local administration. To overcome these issues, various in vivo delivery platforms have been introduced. Here we provide an overview for three different platform technologies for the in vivo delivery of therapeutic siRNAs (siRNA–GalNAc conjugate, SAMiRNA technology, and LNP-based delivery method) and their applications in the treatment of various diseases. In addition, a brief introduction to some rare diseases and mechanisms of siRNA therapeutics-mediated treatment is described.     

Recent in vivo advances in cell-penetrating peptide-assisted drug delivery

ABSTRACT

Introduction: Delivery of macromolecular drugs is an important field in medical research. However, macromolecules are usually unable to cross the cell membrane without the assistance of a delivery system. Cell penetrating peptides (CPPs) are unique tools to gain access to the cell interior and deliver a bioactive cargo into the cytosol or nucleus. In addition to macromolecular delivery, CPPs have been used to deliver smaller bioactive molecules. Therefore CPPs have become an intensive field of research for medical treatment.

Areas covered: In this review, we highlight studies that include CPP in vivo disease models. We review different strategies and approaches that have been used, with specific attention on recent publications. The approaches that have been used include CPP–cargo covalent conjugation strategies and nanoparticle strategies. Various additional strategies have been used to achieve disease targeting, including active targeting, passive targeting, and combined active/passive strategies. As a result, delivery of various types of molecule has been achieved, including small drug molecules, proteins and nucleic acid-based macromolecules (e.g. siRNA, antisense nucleotides and plasmid DNA).

Expert Opinion: Despite recent advances in the field, confusions surrounding CPP internalization mechanisms and intracellular trafficking are hindering the development of new and more efficient vectors. Nevertheless, the recent increase in the number of publications containing in vivo CPP utilization looks promising that the number of clinical trials would also increase in the near future.     

细胞膜穿透肽在肿瘤治疗中的应用策略

目的探讨细胞膜穿透肽(CPP)作为载体在生物大分子药物给药过程中存在的不足,对其解决的方法进行总结。方法分析细胞膜穿透肽的体内给药过程,总结在各个阶段中细胞膜穿透肽的状态、功能及存在的问题。结果将细胞膜穿透肽的给药过程分为3个阶段:体内循环阶段、跨膜阶段及胞内阶段,细胞膜穿透肽要在体内循环阶段保持被掩盖状态,到靶组织细胞附近释放进行跨膜,而在进入细胞后要逃脱溶酶体的降解。结论细胞膜穿透肽能够有效地将生物大分子导入动物细胞,使生物大分子在肿瘤治疗中的应用更具潜力,但仍存在很多的不足之处,需进一步改进。     

Endocytosis and cationic cell-penetrating peptides--a merger of concepts and methods

With the identification of fixation as a major source of artefacts in the cell biological research on cell-penetrating peptides (CPPs), the past two years have witnessed a dramatic development in the CPP field. At least for some of these molecules, endocytosis is now considered to be the major if not the exclusive route of cellular import. However, endocytosis comprises a variety of different pathways with very different implications for the delivery of bioactive molecules to the cytoplasm and nucleus. The endocytosis of CPPs is governed by complex mechanisms similar to those responsible for the internalization of other molecules. Therefore the investigation of uptake and intracellular trafficking of CPPs can benefit enormously from the understanding of the endocytic machinery as well as from the tools that have already been developed for the analysis of endocytosis. This review will introduce aspects of endocytosis relevant to the analysis of CPPs. In addition to the methods, that have already been used for the analysis of CPP trafficking, we will also present other tools and approaches which can be helpful for the research of CPP uptake. Furthermore this review will analyze and summarize recent data providing new insights in endocytosis and intracellular trafficking of CPPs.     

Ultrasound-responsive nanobubbles contained with peptide–camptothecin conjugates for targeted drug delivery

To improve the targeting delivery efficiency of anticancer drug to tumor sites, a new strategy combining cell-permeable peptide (CPP) and ultrasound was reported in this article. In this study, we devised and tested a strategy for functional payload delivery to cells by loading CPP–camptothecin conjugate (CPP–CPT) into nanobubble (CPP–CPT?NB). Here, CPP existing in the conjugation form of CPP and CPT was hidden in nanobubble to cloak the penetration activity of CPP. Meanwhile, local tumor ultrasound was utilized to achieve specific targeting of CPP–CPT to the tumor cells. The mean particle size of the prepared CPP–CPT?NB was ～200?nm, and the drug entrapment efficiency was >80%. Stimulated by ultrasound, over 90% of the entrapped CPP–CPTs would release from the nanobubbles. Subsequent research demonstrated that the CPP–CPT?NB showed effective cellular uptake and significant cytotoxic activity in HeLa cells in vitro. Additionally, after systemic administration in mice, CPP–CPT?NB with ultrasound showed a higher tumor inhibition effect in nude mice xenografted HeLa cells tumors and excellent body safety when compared with normal CPT injection group. In conclusion, the carrier constructed in this study would be a safe and efficiently drug delivery system for specific cancer treatment.     

CNS Delivery Via Adsorptive Transcytosis

Adsorptive-mediated transcytosis (AMT) provides a means for brain delivery of medicines across the blood-brain barrier (BBB). The BBB is readily equipped for the AMT process: it provides both the potential for binding and uptake of cationic molecules to the luminal surface of endothelial cells, and then for exocytosis at the abluminal surface. The transcytotic pathways present at the BBB and its morphological and enzymatic properties provide the means for movement of the molecules through the endothelial cytoplasm. AMT-based drug delivery to the brain was performed using cationic proteins and cell-penetrating peptides (CPPs). Protein cationization using either synthetic or natural polyamines is discussed and some examples of diamine/polyamine modified proteins that cross BBB are described. Two main families of CPPs belonging to the Tat-derived peptides and Syn-B vectors have been extensively used in CPP vector-mediated strategies allowing delivery of a large variety of small molecules as well as proteins across cell membranes in vitro and the BBB in vivo. CPP strategy suffers from several limitations such as toxicity and immunogenicity--like the cationization strategy--as well as the instability of peptide vectors in biological media. The review concludes by stressing the need to improve the understanding of AMT mechanisms at BBB and the effectiveness of cationized proteins and CPP-vectorized proteins as neurotherapeutics.     

Cell‐Penetrating Peptides as Carriers for Oral Delivery of Biopharmaceuticals

Oral delivery of biopharmaceuticals, for example peptides and proteins, constitutes a great challenge in drug delivery due to their low chemical stability and poor permeation across the intestinal mucosa, to a large extent limiting the mode of administration to injections, which is not favouring patient compliance. Nevertheless, cell‐penetrating peptides (CPPs) have shown promising potential as carriers to overcome the epithelium, and this minireview highlights recent knowledge gained within the field of CPP‐mediated transepithelial delivery of therapeutic peptides and proteins from the intestine. Two approaches may be pursued: co‐administration of the carrier and therapeutic peptide in the form of complexes obtained by simple bulk mixing, or administration of covalent conjugates demanding more advanced production methodologies. These formulation approaches have their pros and cons, and which is to be preferred depends on the physicochemical properties of both the specific CPP and the specific cargo. In addition to the physical epithelial barrier, a metabolic barrier must be overcome in order to obtain CPP‐mediated delivery of a cargo drug from the intestine, and a number of strategies have been employed to delay enzymatic degradation of the CPP. The mechanisms by which CPPs translocate across membranes are not fully understood, but possibly involve endocytosis as well as direct translocation, and the CPP‐mediated transepithelial delivery of cargo drugs thus likely involves similar mechanisms for the initial membrane interaction and translocation. However, the mechanisms responsible for transcytosis of the cargo drug, if taken up by an endocytic mechanism, or direct translocation across the epithelium are so far not known.     

Critical issues in delivery of RNAi therapeutics in vivo

RNA interference (RNAi) is a fundamental mechanism of gene regulation and has been harnessed to produce a new class of drugs for treatment of various diseases. A key issue in these applications is how to effectively deliver RNAi therapeutics into target cells. This review is focused on advances in RNA delivery in vivo. To achieve it, novel strategies have been developed to enhance stability of RNA in cells and tissues, overcome barriers to transport of RNA or its carriers in the body, and reduce immunogenicity and cytotoxicity of treatment. Approaches to RNA delivery are divided into three categories in this review: biological, chemical, and physical. Advantages and disadvantages of each method are discussed. At present, effective delivery of RNAi therapeutics in vivo is still a challenge although significant advances have been made in this field.     

Photolabile-caged peptide-conjugated liposomes for siRNA delivery

Light is an ideal general triggered signal, which occurs as a result of its non-invasive nature, desirable controllability and high spatial resolution. However, due to its low penetrability and ability to harm tissues, the use of ultraviolet (UV) light for triggered nanocarrier release in in vivo applications has been limited. Compared with UV light, near-infrared (NIR) light deeply penetrates tissues and is less damaging to cells. In this study, we have devised and tested a strategy for site-specific delivery of small interfering RNA (siRNA) into cancer cells by using liposomes bearing a photolabile-caged peptide (PCP). The positive charges of the lysine residues on the cell-penetrating peptide (CPP) were temporarily caged by the NIR two-photon excitation-responsive protective groups (PG), thereby forming a PCP. Once illuminated by NIR light at tumor tissues, these PGs were cleaved; the positively charged CPP regained its activity and facilitated rapid intracellular delivery of the liposomes into cancer cells. The PCP was connected with 1,2-distearoyl-sn-glycero-3-phosphoethanolamine via a polyethylene glycol (PEG) spacer to prepare the modified liposomes (PCP-LP). Subsequent research demonstrated that the application of the PCP modifications may provide an approach for the selectively targeted delivery of siRNA.     

Current status of gene delivery and gene therapy in lacrimal gland using viral vectors

Gene delivery is one of the biggest challenges in the field of gene therapy. It involves the efficient transfer of transgenes into somatic cells for therapeutic purposes. A few major drawbacks in gene delivery include inefficient gene transfer and lack of sustained transgene expression. However, the classical method of using viral vectors for gene transfer has circumvented some of these issues. Several kinds of viruses, including retrovirus, adenovirus, adeno-associated virus, and herpes simplex virus, have been manipulated for use in gene transfer and gene therapy applications. The transfer of genetic material into lacrimal epithelial cells and tissues, both in vitro and in vivo, has been critical for the study of tear secretory mechanisms and autoimmunity of the lacrimal gland. These studies will help in the development of therapeutic interventions for autoimmune disorders such as Sj?gren's syndrome and dry eye syndromes which are associated with lacrimal dysfunction. These studies are also critical for future endeavors which utilize the lacrimal gland as a reservoir for the production of therapeutic factors which can be released in tears, providing treatment for diseases of the cornea and posterior segment. This review will discuss the developments related to gene delivery and gene therapy in the lacrimal gland using several viral vector systems.