抗肿瘤药物筛选中MTT法和SRB法的比较

在抗肿瘤药物的体外筛选中 ,MTT法和 SRB法是常用的两种方法。我们用MTT法和 SRB法分别测定 3种已知植物抗癌药对 2 2株人肿瘤细胞的抗癌活性 ,对这两种方法进行了详细的比较。通过分析两种方法测出的细胞存活率 ( T/ C)的差异分布和相关系数以及 IC50 的二变量分布 ,比较了两种方法测定结果的异同 ;通过两种方法重复测定 3种药物对 7株人癌细胞的抗癌活性 ,比较了两种方法的重复性 ;通过分析两种方法测定结果 T/ C值随时间变化的程度 ,比较了两种方法测定结果的稳定性。实验结果表明 :MTT法和 SRB法的相关性较好 ,都可用于抗肿瘤药物的体外筛选 ,SRB法更适合于大规模筛选 ,3种抗癌药物的测定结果与临床资料基本一致。     

一种新的以影像为基础的细胞增殖和细胞毒性检测方法

3-(4,5-二甲基-2-噻唑)-2,5-二苯基溴化四唑盐)（MTT）比色法是传统上检测细胞增殖和细胞毒性的常用方法.
CloneSelectTM成像系统是一种以影像为基础的用于分析细胞生长的可视检测系统.本研究采用人结直肠癌HCT116细胞系,运用CloneSelect成像系统和MTT方法分别检测药物阿的平的细胞毒性,并采用Bland Altman作图法比较两种实验方法获得的pEC50值,分析两种研究方法获得的结果的一致性. 结果表明,CloneSelectTM成像系统和MTT法获得的pEC50值具有较好的一致性.与MTT方法相比,基于影像的CloneSelectTM成像分析技术检测快速、无损伤且结果更准确,获取资料不损伤细胞,允许后续其它时间点或动力学检测. 研究提示,这种新的以影像为基础的检测技术可以替代MTT方法,用于分析不同药物的抗细胞增殖活性.     

测量血流量的新方法

日本Waseda大学科学工程学院化学系一个研究组研制成一种测量血流量的新仪器。这种新装置不影响血管压力。而是依靠旋转于血管周围一个畸变传感器来测量,从测量数据中来计算出血流量。这种方法能进行活体测试,而且测量结果准确,费用较低。 血液进入实质脏器的量是十分重要的,它可对各种严重疾病提供适当治疗的依据。目前探测血流量有两种方法,一是采用导管插入血管的方法,二是应用激光或超声多普     

国家基本药物遴选的方法遴选

目的 2009—2011年我国初步建立基本药物制度,对广大公立医院的药品管理和运作影响巨大,但是目前相关的方法体系研究却很少;药物遴选是新医改向纵深推进中实施国家药物政策的起点、重点、难点,旨在系统评价和分析各种方法,完善基本药物的遴选。方法 基于“结构—过程—结果”的指标体系,设计了评分模型,定量比较。结果 量化排序为：背包模型法、专家委员会、循证评价法、层次分析法、专家库抽组、经济学方法。结论 总结了每种方法的特点,并给出了各种方法使用的特定条件和建议。     

血管管腔形成的遗传调控

血管系统是脊椎动物机体中最早发育并行使重要生理功能的复杂系统。血管管腔形成启始于心脏开始跳动和所有其他组织器官形成之前,对于促进血管系统的精确建成和有效灌流,通过营养运输、气体交换和代谢废物清除以确保所有组织器官的形成和生长至关重要。在血管发育过程的两个阶段,即血管发生(vasculogenesis)和血管形成(angiogenesis)中,有效管腔的形成和维持均是保证血管正常发育并行使生理功能的关键环节。血管管腔形成大致可以分为两个关键环节:血管管腔形成的诱导以及内皮细胞极性的建立和维持。重点依据体内遗传修饰模式生物方面的研究结果,从上述两个环节分别阐述血管管腔形成的遗传调控机制。     

两种不同的静脉注射方法对血管的影响

目的：比较两种给药方法对血管损伤的程度。方法：采用自身对照法进行注射泵（实验组）和手推（对照组）静脉注射,指压止血,光镜下观察局部血管及周围组织的变化。结果：注射泵指压止血时间较短,静脉炎发生率低（P〈0.01）,差异具有高度显著性。结论：注射中血流动力学的改变可加重血管的损伤,使指压止血时间延长。     

报告一种同时显示肥大细胞、血管、神经等组织成分的新染色方法

目的为了同时观察组织切片内肥大细胞、血管、神经等组织成分彼此间的形态学关系和联系。方法取大鼠后肢皮肤组织,经恒冷箱切片机切片,然后将组织切片先后经亚铁氰化铜法示乙酰胆碱酯酶和甲苯胺蓝染液染色。结果可同时在组织切片观察到肥大细胞、血管和乙酰胆碱酯酶阳性神经纤维等,以及它们彼此形成的接触联系。结论实验建立的这种新染色方法是显示组织内肥大细胞与血管和神经发生形态学联系的有效方法。     

新型复合血管制备中内皮细胞培养的初步研究

目的:为制备新型复合血管,对内皮细胞的体外培养进行研究。材料和方法:新生儿脐带静脉48条,分别应用酶消化法,贴壁法和机械刮取法获得内皮细胞,以含20％的胎牛血清M199液进行培养,相差镜下观察其贴壁和生长规律。结果:以酶灌注法优于内膜刮取法和内膜贴壁法;而酶灌注法以消化15至20分钟获取细胞和贴壁较多,采取两次消比法较一次消化法效果为佳。结论:三种方法获取和培养的内皮细胞,在数量上用于制备新型复合血管受到一定限制,有关研究有待进一步探讨。     

静脉全麻联合超声引导下前锯肌平面+肋间神经阻滞应用于胸腔镜肺楔形切除术效果分析

摘要 目的：探讨静脉全麻联合超声引导下前锯肌平面+肋间神经阻滞应用于胸腔镜肺楔形切除术效果。方法：选择2021年10月至2022年12月来我院诊治的60例行胸腔镜肺楔形切除术患者,根据随机数字表法,将60例患者分为对照组（30例）与观察组（30例）,对照组30例患者行全麻联合胸椎旁阻滞+肋间神经阻滞的麻醉方法,观察组30例患者行全麻联合前锯肌＋肋间神经阻滞的麻醉方法。对比两组患者监测入室时（T0）、插管即刻（T1）、手术切皮时（T2）、拔管即刻（T3 ）时的平均动脉压及心率,对比两组患者术后2 h、4 h、12 h、24 h、48 h的静息、咳嗽状态下的疼痛评分,对比两组患者T0-T3点的应激反应指标,对比两组患者术中血管活性药的使用剂量,对比两组患者围术期的不良反应发生率。结果：与T0点相比,观察组在T1、T2、T3时的平均动脉压、心率均有明显增加（P<0.05）,而在T1点时,两组比较无差异（P>0.05）;T2、T3点时观察组的平均动脉压、心率明显较对照组低（P<0.05）。随着术后时间延长,两组静息、咳嗽状态下的疼痛评分明显降低（P<0.05）,而同时间点组间对比无统计学意义（P>0.05）。与T0点相比,观察组在T1、T2、T3时的肿瘤坏死因子、白介素6、白介素10水平均有明显增加（P<0.05）,而同时间点观察组与对照组对比无统计学意义（P>0.05）。观察组术中血管活性药的使用剂量明显较对照组低（P<0.05）。观察组的不良反应发生率16.77%低于对照组23.33%,但组间对比无统计学意义（P>0.05）。结论：行静脉全麻联合超声引导下前锯肌平面+肋间神经阻滞的胸腔镜肺楔形切除术者血流动力学更加稳定,术中所需血管活性药物用量明显降低。     

综述——基于纳米材料的生物检测与医学诊断技术：功能化量子点在肿瘤活体成像中的应用

量子点表面经生物分子或药物分子修饰而具有生物功能．功能化量子点具有独特的光学性质和生物相容性,在生物医学光学诊断和治疗领域具有广泛的应用．本文简要介绍了功能化量子点制备及修饰方法,综合评述了量子点在肿瘤活体诊断和治疗中的应用,包括活体淋巴结成像、血管动态成像、肿瘤成像和抗肿瘤药物示踪等,讨论了功能化量子点在肿瘤活体诊断和治疗中的应用前景以及面临的挑战．     

The DrrAB Efflux System of Streptomyces peucetius Is a Multidrug Transporter of Broad Substrate Specificity

The soil bacterium Streptomyces peucetius produces two widely used anticancer antibiotics, doxorubicin and daunorubicin. Present within the biosynthesis gene cluster in S. peucetius is the drrAB operon, which codes for a dedicated ABC (ATP binding cassette)-type transporter for the export of these two closely related antibiotics. Because of its dedicated nature, the DrrAB system is believed to belong to the category of single-drug transporters. However, whether it also contains specificity for other known substrates of multidrug transporters has never been tested. In this study we demonstrate under both in vivo and in vitro conditions that the DrrAB system can transport not only doxorubicin but is also able to export two most commonly studied MDR substrates, Hoechst 33342 and ethidium bromide. Moreover, we demonstrate that many other substrates (including verapamil, vinblastine, and rifampicin) of the well studied multidrug transporters inhibit DrrAB-mediated Dox transport with high efficiency, indicating that they are also substrates of the DrrAB pump. Kinetic studies show that inhibition of doxorubicin transport by Hoechst 33342 and rifampicin occurs by a competitive mechanism, whereas verapamil inhibits transport by a non-competitive mechanism, thus suggesting the possibility of more than one drug binding site in the DrrAB system. This is the first in-depth study of a drug resistance system from a producer organism, and it shows that a dedicated efflux system like DrrAB contains specificity for multiple drugs. The significance of these findings in evolution of poly-specificity in drug resistance systems is discussed.     

A two-step drug repositioning method based on a protein-protein interaction network of genes shared by two diseases and the similarity of drugs

The present study proposed a two-step drug repositioning method based on a protein-protein interaction (PPI) network of two diseases and the similarity of the drugs prescribed for one of the two. In the proposed method, first, lists of disease related genes were obtained from a meta-database called Genotator. Then genes shared by a pair of diseases were sought. At the first step of the method, if a drug having its target(s) in the PPI network, the drug was deemed a repositioning candidate. Because targets of many drugs are still unknown, the similarities between the prescribed drugs for a specific disease were used to infer repositioning candidates at the second step. As a first attempt, we applied the proposed method to four different types of diseases: hypertension, diabetes mellitus, Crohn disease, and autism. Some repositioning candidates were found both at the first and second steps.     

巨噬细胞装载纳米颗粒用于药物递送

目的:活细胞药物递送系统具有主动靶向至肿瘤部位,防止被免疫系统清除等诸多优势。本文提供了一种巨噬细胞负载纳米颗粒的递送方法,并探讨不同载药量对巨噬细胞的活性以及运动性的影响。方法:通过超声乳化法制备包载阿霉素的DOX@PLGA纳米颗粒。纳米粒度分析仪测量粒径和表面电位,透射电镜观察纳米颗粒形态。将DOX@PLGA纳米颗粒与巨噬细胞共同孵育,即得到负载DOX@PLGA纳米颗粒的巨噬细胞用以药物递送。然后通过CCK-8法、LDH法以及细胞迁移实验检测不同载药量情况下细胞活力水平、细胞损伤程度以及细胞运动性。结果:制备的DOX@PLGA纳米颗粒呈圆形或椭圆形,粒径为109.2±2.3 nm;表面电位为-45.0±2.0 m V;载药量为4.61%。当单个巨噬细胞负载0.15 pg DOX时细胞存活率为:71.5±4.4(%);细胞损伤率为:26.3±1.8(%);迁移率为:61.6±5.7(%)。结论:成功制备巨噬细胞负载DOX@PLGA纳米颗粒的递药系统,载药量适当的情况下载体细胞依然具有良好的活性和运动性。     

The Neuroscience Research Institute at Peking University: A Place for the Solution of Pain and Drug Abuse

Neuroscience research in China has undergone rapid expansion since 1980. The Neuroscience Research Institute of Peking University, one of the most active neuroscience research groups in China, was founded in 1987. Currently, the institute is overseeing four research areas, i.e., (1) pain and analgesia, (2) drug abuse and acupuncture treatment for drug addiction, (3) the mechanism of neurological degenerative disorders, and (4) the role of neuroglia in central nervous system injury. The institute is simultaneously investigating both theoretical and clinical studies. Acupuncture remains the core of research, while pain and drug abuse form the two disciplines.     

结核分枝杆菌的耐药基因及其相关分子机制

结核分枝杆菌原发性和继发性耐药是当前控帝】和治疗结核病面临的重要问题,随着分子遗传学的发展,已经阐明了结核分枝杆菌耐药的分子基础是染色体的突变,影响了药靶本身或激活了药物前体的细菌酶,造成MTB的耐药。本文主要就MTB对其常用药物的耐药机制展开讨论,以便正确认识MTB对不同药物的耐药机制,建立快速检测耐药结核分枝杆菌基因型的分子生物学方法。     

Protein function classification via support vector machine approach

Support vector machine (SVM) is introduced as a method for the classification of proteins into functionally distinguished classes. Studies are conducted on a number of protein classes including RNA-binding proteins; protein homodimers, proteins responsible for drug absorption, proteins involved in drug distribution and excretion, and drug metabolizing enzymes. Testing accuracy for the classification of these protein classes is found to be in the range of 84-96%. This suggests the usefulness of SVM in the classification of protein functional classes and its potential application in protein function prediction.     

Gel formulation containing mixed surfactant and lipids associating with carboplatin

The interaction of amphiphilic molecules such as lipids and surfactants with the hydrophilic drug carboplatin was investigated to identify suitable self-assembling components for a potential gel-based delivery formulation. (1) H-NMR Studies in sodium bis(2-ethylhexyl) sulfosuccinate (aerosol-OT, AOT)-based reverse micelles show that carboplatin associates and at least partially penetrates the surfactant interface. Langmuir monolayers formed by dipalmitoyl(phosphatidyl)choline are penetrated by carboplatin. Carboplatin was found to also penetrate the more rigid monolayers containing cholesterol. A combined mixed surfactant gel formulation containing carboplatin and cholesterol for lymphatic tissue targeting was investigated for the intracavitary treatment of cancer. This formulation consists of a blend of the surfactants lecithin and AOT (1 : 3 ratio), an oil phase of isopropyl myristate, and an aqueous component. The phases of the system were defined within a pseudo-ternary phase diagram. At low oil content, this formulation produces a gel-like system over a wide range of H(2) O content. The carboplatin release from the formulation displays a prolonged discharge with a rate three to five times slower than that of the control. Rheological properties of the formulation exhibit pseudoplastic behavior. Microemulsion and Langmuir monolayer studies support the interactions between carboplatin and amphiphilic components used in this formulation. To target delivery of carboplatin, two formulations containing cholesterol were characterized. These two formulations with cholesterol showed that, although cholesterol does little to alter the phases in the pseudo-ternary system or to increase the initial release of the drug, it contributes significantly to the structure of the formulation under physiological temperature, as well as increases the rate of steady-state discharge of carboplatin.     

湖北地区部分医院肠球菌耐药性监测及相关因素分析

目的：了解湖北地区2000年度肠球菌的分离情况及耐药状况,并对抗感染用药进行探索,指导临床合理用药。方法：对湖北地区15所三甲医院各类临床标本中的538株肠球菌采用API细菌鉴定系统或Vitek全自动细菌鉴定系统进行分离鉴定,用纸片扩散法进行药敏试验,并以WHO细菌耐药监测网提供的WHONET4软件分析系统对试验数据进行分析处理。结果：本试验共分离肠球菌13个种别538株,其中大多数是粪肠球菌408株,占75.8%,屎肠球菌54株,占10.0%;坚忍肠球菌24株,占4.46%;鸟肠球菌10株,占1.86%。酪黄肠球菌6株,占1.11%;母鸡肠球菌6株,占1.11%等。本研究结果表明肠球菌的主要感染部位为泌尿道31.4%和各种分泌物31.0%。同时从药敏结果可见屎肠球菌和坚忍肠球菌对大多数抗生素的耐药性高于粪肠球基本国策 ;并发现糖肽类抗生素耐药菌株比例尚不高;庆大霉素呈高水平耐药球菌株比例则较高;对青霉素和氨卞西林耐药,本研究结果亦有反映,并且鸟肠球菌的耐药性高于粪肠球菌近三倍。结论：本年度分离的肠球菌占全年总分离菌的第六位,说明我国目前由肠球菌引起的感染所占比例不高,但仍应引起临床的高度重视,并进行动态的监测。加强对肠球菌特别是VRR耐药性的监测是非常必要的,泌尿道肠球菌感染的抗生素中呋喃妥因的耐药率较四环素和喹诺酮类药物为低,故仍不失为治疗该类泌感的首选药物,菌株差别与地区有关,由于肠球菌属中的不同种对抗生素的敏感性不同。因此种的鉴定是对该地区医院感染暴发流行,选择治疗方案的重要工具。     

Repurposing approach identifies new treatment options for invasive fungal disease

Drug repositioning is the process of discovery, validation and marketing of previously approved drugs for new indications. Our aim was drug repositioning, using ligand-based and structure-based computational methods, of compounds that are similar to two hit compounds previously selected by our group that show promising antifungal activity. Through the ligand-based method, 100 compounds from each of three databases (MDDR, DrugBank and TargetMol) were selected by the Tanimoto coefficient, as similar to LMM5 or LMM11. These compounds were analyzed by the scaffold trees, and up to 10 compounds from each database were selected. The structure-based method (molecular docking) using thioredoxin reductase as the target drug was performed as a complementary approach, resulting in six compounds that were tested in an in vitro assay. All compounds, particularly raltegravir, showed antifungal activity against the genus Paracoccidioides. Raltegravir, an antiviral drug, showed promising antifungal activity against the experimental murine paracoccidioidomycosis, with significant reduction of the fungal burden and decreased alterations in the lung structure of mice treated with 1 mg/kg of raltegravir. In conclusion, the combination of two in silico methods for drug repositioning was able to select an antiviral drug with promising antifungal activity for treatment of paracoccidioidomycosis.     

院内呼吸科下呼吸道感染病原学及耐药性结果分析

目的分析大连市第五人民医院2009年呼吸科送检1160例次痰标本细菌检查情况,为临床提供感染数据以及合理使用抗生素提供依据。方法按常规方法将临床送检的痰标本进行细菌培养、分离。使用BIO-FOSUN微生物鉴定药敏分析系统进行细菌鉴定及抗生素敏感试验。结果一年来共收痰标本1160例次,检出病原菌253例次,阳性率为21.8%。病原菌依次为肺炎克雷伯菌83株,铜绿假单胞菌57株,乙酸钙不动杆菌29株,阴沟肠杆菌14株,粘质沙雷菌13株,大肠埃希菌10株,,金黄色葡萄球菌5株等。大多数革兰阴性菌对亚胺培南较敏感,对头孢菌素类均有不同程度的耐药。革兰阳性球菌对万古霉素敏感,对其他抗生素有不同程度的耐药。结论该院呼吸科患者下呼吸道感染以革兰阴性杆菌为主,常见病原菌耐药性未呈现高水平、多重耐药情况。提示临床医生应与检验科密切配合,关注细菌感染情况,根据药敏试验结果合理选用抗生素。