学时数较少的药学物理化学教学探索

根据课程学时的限制，结合药学专业课程的要求，对物理化学理论教学内容进行了取舍，强调上好第一课，并对部分内容进行双语授课的尝试，对实验教学也进行了探索。     

对药学无机化学的教学体会

根据药学专业无机化学的学科特点出发，就课堂授课过程中如何改革教学方法、激发学生学习兴趣、设计合理的教学过程等方面进行探讨，以提高学生认知能力和自主学习的欲望，达到善于总结和提高学习效率的目的。     

A probabilistic approach to measure the strength of bone cell adhesion to chemically modified surfaces

Patterned surfaces with alternating regions of amino silanes [N-(2-aminoethyl)-3-aminopropyl-trimethoxysilane (EDS)] and alkyl silanes [dimethyldichlorosilane (DMS)] have been used to alter the kinetics of spatial distribution of cellsin vitro. In particular, we have previously observed the preferential spatial distribution of bone cells on the EDS regions of EDS/DMS patterned surfaces (10). In this study, we examined whether the mechanism of spatial distribution of cells on the EDS regions was adhesion mediated. Homogeneous layers of EDS and DMS were immobilized on quartz substrates and characterized by contact angle, X-ray photoelectron spectroscopy, and spectroscopic ellipsometry. The strength of bone cell attachment to the modified substrates was examined using a radial flow apparatus, within either 20 min or 2 hr of cell incubation in the presence of serum. A Weibull distribution was chosen to characterize the strength of cell-substratum adhesion. Within 20 min of cell exposure, the strength of adhesion was significantly larger on EDS and clean surfaces, compared with DMS surfaces (p<0.0001). Within 2 hr of cell incubation, there was no statistical difference between the strength of cell adhesion to EDS, DMS, and clean surfaces. The results of this study suggest that the surface chemistry mediates adhesion-based spatial cell arrangement through a layer of adsorbed serum proteins.     

Developmental changes of chick cerebellar cortex revealed by monoclonal antibodies

Immunohistochemistry studies of the embryonic and newly hatched chick cerebellum were performed with 13 monoclonal antibodies (MAbs) raised against the embryonic chick optic nerve and a MAb which binds to cell nuclei. Neural MAbs differentially stained Purkinje cells, the external granular layer, molecular layer, internal granular layer, climbing fibers, basket cell axons, Bergmann glia and Ramón y Cajal's ansiform fibers. At the different developmental stages each component responded to MAbs differently. For example, staining of Purkinje cells with MAbs 23C10, 82E10 and 94C2 appeared on day 11 of incubation and disappeared sequentially after day 18. These results reveal molecular heterogeneity not only in cerebellar neurons but also at various developmental stages.     

功能化多壁碳纳米管对血管内皮细胞毒性的初步研究

目的:由于碳纳米管的广泛应用,其对人类及环境造成的影响日益受到人们的关注.血管是碳纳米管进入人体的首要通道之一,因此探讨功能化与原始状态的多壁碳纳米管对血管内皮细胞的细胞毒性作用具有重要意义.方法:自人脐静脉分离血管内皮细胞,免疫组化进行细胞鉴定.选择0.5～1 μm经羧基修饰的功能化多壁碳纳米管(f-CNTs)与同等长度未经修饰的多壁碳纳米管(p-CNTs)制备颗粒悬液,与血管内皮细胞共育后,利用噻唑蓝实验(MTT比色法)检测细胞活力,透射电镜进行细胞形态学观察;制备碳纳米管透析液,通过MTT实验检测其对细胞的毒性作用.结果:①2种纳米管均对原代脐静脉内皮细胞(HUVEC)产生一定的毒性作用,而且其细胞毒性随剂量递增呈上升趋势.与细胞共育24 h后,p-CNTs的毒性大于f-CNTs;而共育48 h及72 h后,f-CNTs毒性大于p-CNTs.②透射电镜实验表明,f-CNTs共育细胞内空腔明显多于p-CNTs,而空腔内f-CNTs聚集体紧密程度明显小于p-CNTs.③对2种多壁碳纳米管透析后的细胞培养基对细胞活力无明显影响.结论:根据上述结果笔者推测.p-CNTs细胞毒性的主导因素是聚集程度,f-CNTs对细胞毒性的主导因素为表面效应.进入细胞的碳纳米管数目增多,造成细胞损伤或阻断细胞内代谢通路,可能是共育后期f-CNTs细胞毒性大于p-CNTs的原因.     

Surface chemistry modulates focal adhesion composition and signaling through changes in integrin binding

Biomaterial surface properties influence protein adsorption and elicit diverse cellular responses in biomedical and biotechnological applications. However, the molecular mechanisms directing cellular activities remain poorly understood. Using a model system with well-defined chemistries (CH₃, OH, COOH, NH₂) and a fixed density of the single adhesive ligand fibronectin, we investigated the effects of surface chemistry on focal adhesion assembly and signaling. Surface chemistry strongly modulated integrin binding and specificity—₅β₁ integrin binding affinity followed the pattern OH>NH₂=COOH>CH₃, while integrin _Vβ₃ displayed the relationship COOH>NH₂OH=CH₃. Immunostaining and biochemical analyses revealed that surface chemistry modulates the structure and molecular composition of cell-matrix adhesions as well as focal adhesion kinase (FAK) signaling. The neutral hydrophilic OH functionality supported the highest levels of recruitment of talin, -actinin, paxillin, and tyrosine-phosphorylated proteins to adhesive structures. The positively charged NH₂ and negatively charged COOH surfaces exhibited intermediate levels of recruitment of focal adhesion components, while the hydrophobic CH₃ substrate displayed the lowest levels. These patterns in focal adhesion assembly correlated well with integrin ₅β₁ binding. Phosphorylation of specific tyrosine residues in FAK also showed differential sensitivity to surface chemistry. Finally, surface chemistry-dependent differences in adhesive interactions modulated osteoblastic differentiation. These differences in focal adhesion assembly and signaling provide a potential mechanism for the diverse cellular responses elicited by different material properties.     

Concurrent infections with vector-borne pathogens associated with fatal anaemia in cattle: haematology and blood chemistry

An outbreak of a fatal haemolytic anaemia in a dairy herd of cattle in Switzerland was shown to be associated with infections with five vector-borne pathogens, namely Anaplasma marginale, A. phagocytophilum, Babesia bigemina, a Theileria spp belonging to the buffeli/sergenti/orientalis complex and haemotrophic Mycoplasma spp. The latter three had not been documented before this outbreak in Switzerland. To characterise the haematological and blood chemical changes in these unique cows, packed cell volume was determined in all 286 blood samples, blood smears, and complete haematology were performed from 285 and 173 blood samples, respectively, and biochemical parameters were assayed in 105 serum samples. Regenerative anaemia was the key sign of illness. Red blood cells of anaemic cattle were hypochromic and macrocytic. Anaemic animals had reduced platelet cell counts and increased total white cell counts. In addition, increased serum bilirubin, blood aspartate aminotransferase, gamma glutamyltransferase, glutamic dehydrogenase and blood urea nitrogen and decreased magnesium, calcium and albumin levels were found in anaemic cattle when compared to animals with normal packed cell volume. Most changes could not be attributed to a single infection. A. marginale seemed to be important in causing the outbreak, but co-infections may have aggravated the disease development and clinical signs. Thus, when encountering cattle with haemolytic anaemia, all of the mentioned pathogens should be included as differential diagnosis.     

Comparative haematological and plasma chemistry values in purpose-bred squirrel, cynomolgus and rhesus monkeys

Haematological and blood chemistry data have been compared for three species of purpose-bred primates, Macaca mulatta (rhesus monkey), Macaca fascicularis (cynomolgus monkey), Saimiri sciureus (squirrel monkey). These species were housed over a 9-month period in identical conditions (diet, tap water, room temperature and relative humidity, and lighting regime) in our primate facility. Blood and urine assays were conducted using the same pre-analytical conditions (blood sampling procedure, anticoagulant and storage of sample), and analytical methods (reagent and equipment).The results indicated that squirrel, rhesus and cynomolgus monkeys have essentially biologically similar values for all of the parameters examined. However, haemoglobin level, reticulocyte, plasma total cholesterol, triglyceride, albumin and urea nitrogen values, and urinary osmolality in cynomolgus monkeys were statistically lower than those of rhesus monkeys. Erythrocyte count, plasma ALT, calcium and potassium in cynomolgus monkeys were statistically higher than those of rhesus monkeys. Erythrocyte count, haemoglobin level, haematocrit, reticulocyte, plasma total bilirubin and chloride and urinary osmolality in cynomolgus monkeys were statistically lower than those of squirrel monkeys. Leucocyte count, plasma total protein, albumin and calcium in cynomolgus monkeys were statistically higher than those of squirrel monkeys. Squirrel monkeys showed marked deviations in four assays from the other two species: ratio of lymphocytes to neutrophils, AST, ALT and urine volume.The results obtained in this study will be used as baseline data for haematology and clinical chemistry characteristics for three species of purpose-bred monkeys.     

功能化多壁碳纳米管对血管内皮细胞毒性的初步研究

目的:由于碳纳米管的广泛应用,其对人类及环境造成的影响日益受到人们的关注.血管是碳纳米管进入人体的首要通道之一,因此探讨功能化与原始状态的多壁碳纳米管对血管内皮细胞的细胞毒性作用具有重要意义.方法:自人脐静脉分离血管内皮细胞,免疫组化进行细胞鉴定.选择0.5～1 μm经羧基修饰的功能化多壁碳纳米管(f-CNTs)与同等长度未经修饰的多壁碳纳米管(p-CNTs)制备颗粒悬液,与血管内皮细胞共育后,利用噻唑蓝实验(MTT比色法)检测细胞活力,透射电镜进行细胞形态学观察;制备碳纳米管透析液,通过MTT实验检测其对细胞的毒性作用.结果:①2种纳米管均对原代脐静脉内皮细胞(HUVEC)产生一定的毒性作用,而且其细胞毒性随剂量递增呈上升趋势.与细胞共育24 h后,p-CNTs的毒性大于f-CNTs;而共育48 h及72 h后,f-CNTs毒性大于p-CNTs.②透射电镜实验表明,f-CNTs共育细胞内空腔明显多于p-CNTs,而空腔内f-CNTs聚集体紧密程度明显小于p-CNTs.③对2种多壁碳纳米管透析后的细胞培养基对细胞活力无明显影响.结论:根据上述结果笔者推测.p-CNTs细胞毒性的主导因素是聚集程度,f-CNTs对细胞毒性的主导因素为表面效应.进入细胞的碳纳米管数目增多,造成细胞损伤或阻断细胞内代谢通路,可能是共育后期f-CNTs细胞毒性大于p-CNTs的原因.     

Cyclodextrins in Polymer Synthesis: Free Radical Polymerization of β‐Cyclodextrin Complexes of Photosensitive Mesoionic 6‐Oxo‐1,6‐dihydropyrimidin‐3‐ium‐4‐olates in Aqueous Medium

Solid mesoionic 2‐[2‐(isopropenylcarbonyloxy)ethylthio]‐1‐methyl‐6‐oxo‐3‐phenyl‐5‐propyl‐1,6‐dihydropyrimidin‐3‐ium‐4‐olate was complexed in water using β‐cyclodextrin (β‐CD) and randomly methylated β‐CD, which resulted in polymerizable complexes with 2:1 stoichiometry. The β‐CD complex was characterized using ¹H NMR, ROESY NMR and UV spectroscopy. Polymerization of the complex prepared from methylated β‐CD led to a photosensitive polymer, which precipitated during polymerization and was nearly free of CD. Polymerization was carried out with a water‐soluble redox initiator. In addition, a copolymer with methyl methacrylate was prepared from the complexes, which showed a different mass‐dependent distribution in the incorporation in comparison to a copolymer prepared without CD in organic solvents.