中药物质基础研究的思路与方法

本文以中药现代化和国际化为导向,论述了中药研究中的重点与难点问题。阐述了中药研究的主要发展方向是突破中药化学物质基础研究,并提出了以活性导向的标准组分模式的中药物质基础研究新思路、新观点。强调中药物质基础研究的关键技术发展的重要性,并介绍了现代中药分离分析与表征的方法与技术。     

琼脂糖凝胶基片用于蛋白质与抗原分子的固定研究与表征

目的 探索一种新型的适用于生物分子偶联的固相载体，并对其用于蛋白质与抗原分子微阵列制作的实验条件和检测结果进行评价。方法 于氨基硅烷化的载玻片表面制作琼脂糖凝胶膜，经NaIO4氧化和戊二醛分子偶联后，得到3种不同的凝胶表面，分别考察其固定生物分子的偶联效率、检测敏感度和稳定性能，并用原子力显微术和X射线光电子能谱对其表面进行表征和对比研究。结果 未经氧化的琼脂糖凝胶基片固定生物分子的能力和检测敏感度低于经NaIO4氧化与后续醛化处理的基片，其中又以后者的应用效果最佳，温度因素（实验温度分别为37℃、25℃和4℃）对用此法制作的蛋白质与抗原分子微阵列的使用无显著影响。表征结果提供了琼脂糖凝胶表面固定生物分子的直接证据和内在机制。结论 经修饰处理的琼脂糖凝胶基片可有效固定不同的生物分子，用其作为固相表面制作的蛋白质与抗原分子微阵列可取得较为理想的实验效果。     

两种普朗尼克材料联合聚氰基丙烯酸正丁酯载紫杉醇的控药释放能力

背景：包含普朗尼克P123的载紫杉醇聚合物胶束能够有效的延长药物体内循环时间,并且能够改变紫杉醇的作用靶位.但是,这种紫杉醇聚合物胶束在溶液中的稳定性以及载药能力仍有待提高. 目的：观察载紫杉醇聚氰基丙烯酸正丁酯-普朗尼克P123/F68胶束的药剂学特性和体外抗肿瘤能力.方法：采用薄膜水化法,以聚氰基丙烯酸正丁酯为交联剂,普朗尼克P123/F68为载体材料,制备载疏水性药物-紫杉醇纳米胶束.应用透射电镜观察胶束形态;电位粒度分析仪测定胶束电位和粒径;高效液相色谱分析方法测定胶束载药量和包封率;荧光探针法测定胶束临界胶束浓度;体外试验考察胶束的释药情况、稳定性以及抗肿瘤情况. 结果与结论：实验制备的载药胶束为圆形,粒径和电位分别在100 nm和-10 mV左右,包封率和载药量为（93.3±2.15）%和（1.82±0.04）%,临界胶束浓度为0.067 g/L.药物体外释放试验和稳定性试验显示,该载药胶束具有一定的缓释功能和抗稀释能力.MTT试验结果表明,与游离药物相比,载药胶束具有更强的杀伤乳腺癌细胞MCF-7的能力.可见,载紫杉醇聚氰基丙烯酸正丁酯-普朗尼克P123/F68胶束具有明显的控制药物释放的能力和良好的稳定性,抗肿瘤能力强.     

Immunomodulatory treatment strategies in inflammatory cardiomyopathy: current status and future perspectives

Chronic autoimmunity and viral persistence constitute prognostic factors for adverse outcome in dilated cardiomyopathy patients. Inflammatory cardiomyopathy is a specific cardiomyopathy entity diagnosed in approximately 50% of dilated cardiopmyopathy patients by immunohistological quantification of immunocompetent infiltrates and cell adhesion molecule abundance. Patients with autoimmune inflammatory cardiomyopathy benefit from immunosuppressive treatment and immunoadsorption by improvement of left ventricular ejection fraction and heart failure symptoms, paralleled by a significant suppression of intramyocardial inflammation. However, dilated cardiomyopathy patients with viral persistence do not respond favorably to immunosuppression.     

竹荪子实体多糖的提取及化学组成

从天然竹荪中经脱脂、采用20、40、60、80℃四个不同温度热水梯度抽提、Sevage法脱蛋白、乙醇沉淀得到四种粗竹荪多糖。对其中在60℃下提取的多糖进行研究。经测定此多糖具有较高的粘度,再对其用丙酮进行分级得到纯度较高的竹荪多糖-3。用气相色谱对其单糖组成结构进行分析,结果表明荪多糖-3是一种以葡萄糖为主的多糖,通过刚果红实验证实此多糖可能具有三螺旋结构。     

复方丹参脉冲片的制备工艺及体外释药特性研究

目的:根据冠心病发病的时辰节律性,以复方丹参方的药效物质和作用机制为基础,制备复方丹参脉冲片并考察其体外释药特性.方法:脉冲片由含有复方丹参药物、丁二酸、羧甲基淀粉钠、乳糖和微晶纤维素的片芯外包3层衣膜构成,内层为隔离层,由Eudragit RL 100构成;中间层为溶胀层,由HPMC E5构成;外层为控释层,由Eudragit RS 100,RL 100和EC的混合物构成.结果:5种有效成分丹参素、原儿茶醛、人参皂苷Rg_1,Rb_1、三七皂苷R_1以相近的释药规律释药,脉冲片在膨胀作用的基础上,利用渗透泵、有机酸诱导和扩散机理在设定的时滞后快速释药.结论:成功制得一定时滞后快速释放的复方丹参脉冲片用于冠心病的防治,可减少"心血管事件"发生的几率.     

Polyhydroxyalkanoates (PHAs) as Biomaterials in Tissue Engineering: Production,Isolation, Characterization

Polyhydroxyalkanoates (PHAs) are biodegradable and biocompatible biopolymers. These biomaterials have grown in importance in the fields of tissue engineering and tissue reconstruction for structural applications where tissue morphology is critical, such as bone, cartilage, blood vessels, and skin, among others. Furthermore, they can be used to accelerate the regeneration in combination with drugs, as drug delivery systems, thus reducing microbial infections. When cells are cultured under stress conditions, a wide variety of microorganisms produce them as a store of intracellular energy in the form of homo- and copolymers of [R]—hydroxyalkanoic acids, depending on the carbon source used for microorganism growth. This paper gives an overview of PHAs, their biosynthetic pathways, producing microorganisms, cultivation bioprocess, isolation, purification and characterization to obtain biomaterials with medical applications such as tissue engineering.     

人乳头状瘤病毒86E7重组蛋白的高效表达、纯化和表征

[目的]基因工程法构建谷胱甘肤S-转移酶(GST)-人乳头状瘤病毒(Human papillomavirus)HPV86 E7融合蛋白表达质粒,高效表达、纯化、鉴定蛋白质和分析蛋白质的存在形式.[方法]GST-HPV86 E7融合蛋白基因与pGEX-6P-1载体连接成重组表达质粒pGEX-6P-1-GST-HPV86 E7,重组质粒转入BL21大肠杆菌,经IPTG诱导表达融合蛋白GST-HPV86 E7,柱上切除GST标签,Glutathione Sepharose 4B亲和层析纯化蛋白,SDS-PAGE、MALDI-TOF和色谱质谱联用系统分别用于蛋白质纯度分析、分子量测定和蛋白鉴定;非变性电泳和分子筛排阻色谱用于蛋白质四级结构的分析.[结果]HPV86 E7大肠杆菌中正确表达,基质辅助激光解析飞行时间质谱测得的HPV86 E7精确分子量为(11319.76±5.66)Da.反相高压液相色谱-电喷雾串联质谱鉴定的匹配肽段有11个肽段,氨基酸的覆盖率为90%.非变性PAGE和凝胶过滤层析实验观察到了HPV86 E7的单聚体、二聚体及多聚体.[结论]重组质粒pGEX-6P-1-GST-HPV86 E7成功构建,诱导后高效表达GST-HPV86 E7融合蛋白,HPV86 E7得到纯化,质谱分析证实表达蛋白为86E7蛋白,以多聚体形式存在.     

Comparison of Testing Methods for Evaluating the Resistance of Alkali-Activated Blast Furnace Slag Systems to Sulfur Dioxide

Alkali-activated systems (AAS) represent an ecologically and economically sustainable inorganic binder as an alternative to ordinary Portland cement (OPC). One of the main benefits of AAS is their durability in aggressive environments, which can be equal or even better than that of OPC. In this paper, the influence of the type of alkaline activator in alkali-activated blast furnace slag (AAS) in terms of resistance to sulfur dioxide corrosion was investigated. The durability testing process was based on the CSN EN ISO 3231 standard and simultaneously compared with mortar samples prepared by using Blastfurnace cement CEM III/A 32.5R. The degradation progress was evaluated by employing several different methods such as observing the compressive strength development, weight change evaluation, non-destructive testing methods like ultrasound or impact echo technique, or visual phenolphthalein technique. Subsequently, fundamental characterization of samples by the XRD method was performed during the degradation test. The obtained results indicate that none of the testing methods used could be prioritized over others to determine the resistance of AAS against the action of sulfur dioxide. For this reason, the durability testing of AAS remains an issue, and the development of specific standards considering the behavior of AAS seems necessary.     

Hematopoietic reconstitution after bone marrow transplantation: Assessment with MR imaging and H-1 localized spectroscopy

Magnetic resonance (MR) studies were performed in 14 patients as early as possible (21–110 days) after bone marrow transplantation (BMT). MR characteristics of lumbar vertebral bone marrow were studied with T1-weighted spin-echo imaging, water- and fatselective imaging with a frequency-selective excitation technique, and point-resolved spatially localized proton spectroscopy. Signals from water and fat protons and their T1 and T2 values were analyzed. Water proton signal intensity correlated well with cellularity within bone marrow, as determined with parallel iliac crest biopsies. The fraction of signal from water in red bone marrow of patients with allogeneic transplants from siblings (four cases) was significantly higher than in four patients with autologous transplants. The latter showed very low cellularity in the period of about 4 weeks after BMT because of the cytotoxic pretreatment of the bone marrow. The MR results in six patients with allogeneic transplants from unrelated donors ranged widely, depending on the complications after BMT. Analysis of data obtained with the different techniques showed that water- and fat-selective MR imaging and spectroscopic methods are useful for noninvasive monitoring of hematopoietic reconstitution after BMT.