延胡索乙素与白芷总香豆素、总挥发油配伍药效学比较研究

目的：评价延胡索乙素、白芷总香豆素及总挥发油配伍的药效学比较研究。方法：小鼠醋酸扭体法、热板镇痛法、甩尾法三种模型比较元胡止痛片（A组）、延胡索乙素（B组）、白芷总香豆素（C组）、白芷总挥发油（D组）、延胡索乙素与白芷总香豆素配伍（E组）、延胡索乙素与白芷总挥发油配伍（F组）及延胡索乙素与白芷总香豆素、白芷总挥发油配伍（G组）后的镇痛作用。结果：A、G组镇痛效果最佳,B、E、F组具显著的镇痛作用,C、D组与模型组相比,有一定的镇痛作用,但较弱。结论：延胡索乙素与白芷总香豆素、总挥发油配伍使用,具有显著的镇痛作用,类似于元胡止痛片中元胡白芷的配伍作用,但成分更为明确,标准可控,使用量小,属于分子中药组方范畴。元胡的主要有效成分延胡索乙素与白芷的主要成分白芷总香豆素、总挥发油均具有镇痛作用,符合中药元胡及白芷的功效,本实验采用药物有效成分进行配伍,成分明确,结果可控。     

Intravenous Microinjections of Zebrafish Larvae to Study Acute Kidney Injury

In this video article we describe a zebrafish model of AKI using gentamicin as the nephrotoxicant. The technique consists of intravenous microinjections on 2 dpf zebrafish. This technique represents an efficient and rapid method to deliver soluble substances into the bloodstream of zebrafish larvae, allowing for the injection of 15-20 fish per hour. In addition to AKI studies, this microinjection technique can also be used for other types of experimental studies such as angiography. We provide a detailed protocol of the technique from equipment required to visual measures of decreased kidney function. In addition, we also demonstrate the process of fixation, whole mount immunohistochemistry with a kidney tubule marker, plastic embedding and sectioning of the larval zebrafish. We demonstrate that zebrafish larvae injected with gentamicin show morphological features consistent with AKI: edema, loss of cell polarity in proximal tubular epithelial cells, and morphological disruption of the tubule.     

Glycosylation engineering of therapeutic IgG antibodies: challenges for the safety,functionality and efficacy

Glycosylation of the Fc region of IgG has a profound impact on the safety and clinical efficacy of therapeutic antibodies. While the biantennary complex-type oligosaccharide attached to Asn297 of the Fc is essential for antibody effector functions, fucose and outer-arm sugars attached to the core heptasaccharide that generate structural heterogeneity (glycoforms) exhibit unique biological activities. Hence, efficient and quantitative glycan analysis techniques have been increasingly important for the development and quality control of therapeutic antibodies, and glycan profiles of the Fc are recognized as critical quality attributes. In the past decade our understanding of the influence of glycosylation on the structure/function of IgG-Fc has grown rapidly through X-ray crystallographic and nuclear magnetic resonance studies, which provides possibilities for the design of novel antibody therapeutics. Furthermore, the chemoenzymatic glycoengineering approach using endoglycosidase-based glycosynthases may facilitate the development of homogeneous IgG glycoforms with desirable functionality as nextgeneration therapeutic antibodies. Thus, the Fc glycans are fertile ground for the improvement of the safety, functionality, and efficacy of therapeutic IgG antibodies in the era of precision medicine.     

Cloning and characterization of novel tumor-targeting immunocytokines based on murine IL7

We generated and characterized novel antibody-cytokine fusion proteins (“immunocytokines”) based on murine interleukin-7 (IL7), an immunomodulatory protein which has previously shown anti-cancer activity in preclinical models and whose human counterpart is currently being investigated in clinical trials. The sequential fusion of the clinical-stage antibody fragment scFv(F8), specific to a tumor-associated splice isoform of fibronectin, yielded an immunocytokine (termed “F8-mIL7”) of insufficient pharmaceutical quality and in vivo tumor targeting performance, with a striking dose dependence on tumor targeting selectivity. By contrast, a novel immunocytokine design (termed “F8-mIL7-F8”), in which two scFv moieties were fused at the N- and C-terminus of murine IL7, yielded a protein of excellent pharmaceutical quality and with improved tumor-targeting performance [tumor: blood ratio = 16:1, 24 h after injection]. Both F8-mIL7 and F8-mIL7-F8 could induce tumor growth retardation in immunocompetent mice, but were not able to eradicate F9 tumors. The combination of F8-mIL7-F8 with paclitaxel led to improved therapeutic results, which were significantly better compared to those obtained with saline treatment. The study indicates how the engineering of novel immunocytokine formats may help generate fusion proteins of acceptable pharmaceutical quality, for those immunomodulatory proteins which do not lend themselves to a direct fusion with antibody fragments.     

静脉与硬膜外注射地塞米松对吗啡硬膜外术后镇痛的影响

目的:比较两种不同途径注射地塞米松磷酸钠对吗啡硬膜外术后镇痛的影响。方法:选择200例(ASAⅠ-Ⅱ)在腰硬联合麻醉下行腹式子宫切除术的患者,随机分为A、B、C、D四组(n=50),各组均给以硬膜外注射2.5 mg吗啡作为术后镇痛治疗的同时,A组静脉注射安慰剂(生理盐水),B组静脉注射地塞米松磷酸钠10 mg,C组静脉注射地塞米松磷酸钠5 mg,D组硬膜外注射地塞米松磷酸钠5 mg及静脉注射安慰剂(生理盐水),以上均以5 mL作为注射容积。观察和比较术后24 h内各组恶心和呕吐(PONV)、皮肤瘙痒、补救镇痛、呼吸抑制的发生率、排气时间和补救镇痛时间。结果:B、C、D三组的PONV总发生率显著低于A组(P0.0083),而B、C、D三组之间比较无显著差异(P0.0083);A、B、C、D四组间恶心的发生率无显著差异(P0.05),而D组呕吐的发生率明显低于A组(P0.0083);B组皮肤瘙痒的发生率明显低于A组(P0.0083);四组患者的VAS评分比较无显著差异,均达到满意的镇痛效果(P0.05)。四组患者补救镇痛的发生率、补救镇痛药量和排气时间比较无明显差异(P0.05),而C、D组的补救镇痛时间明显比A组延长(P0.0083),四组患者均未出现呼吸抑制。结论:地塞米松磷酸钠可降低吗啡硬膜外术后恶心和呕吐的发生率,延长补救镇痛时间;硬膜外注射地塞米松磷酸钠对降低呕吐的发生率更有效;静脉注射地塞米松磷酸钠10 mg可降低瘙痒的发生率,且无明显的不良反应。     

自由体位分娩加分娩减痛法在初产顺产妇中的临床应用效果

目的:研究和探讨自由体位分娩法和分娩减痛法结合起来在初产顺产妇中的临床应用效果。方法:选取2013年1月至2013年12月已被收治的符合标准的初产顺产妇女360例为研究对象,将其随机分成A1组(自由体位分娩法组)、A2组(分娩减痛法组)和A1+A2组(两种方法结合组),每组120例。观察和比较三组产妇阴道分娩产程时间、疼痛等级评分、出血量、剖宫产率、新生儿Apgar评分情况和产后不良情况发生率。结果:①A1+A2组的分娩产程时间低于A1组及A2组,疼痛等级评分优于A1组和A2组(P均0.05);②A1+A2组的分娩过程中的出血量和剖宫产率均低于A1组及A2组(P0.05);③A1+A2组新生儿Apgar评分情况比A1组及A2组好(P0.05);④A1+A2组产后不良情况的发生率也低(P0.05)。然而以上观察指标A1组与A2组之间的差异均无统计学意义(P0.05)。结论:在临床实践过程中对初产顺产产妇实施自由体位分娩结合分娩减痛法可以显著地改善产妇分娩过程中的不良情况,对新生儿的影响也是积极的。两种方法的综合应用更具有良好的临床效果。     

Anti-A and anti-B haemagglutinin levels in intravenous immunoglobulins: Are they on the rise? A comparison of four different analysis methods and six products

Recent reports of severe haemolytic reactions upon high dose treatment with new generation intravenous immunoglobulins (IVIGs) prompted us to examine the anti-A and anti-B haemagglutinin content of these therapeutics. We compared four different test methods, namely the indirect and direct haemagglutination test as described in the European Pharmacopoiea (Ph. Eur.) and two commercial gelcard systems with the aim to define the most reliable method for a large-scale comparison of different IVIG products. Absolute titres varied when the same samples were analyzed by the four methods, while the relative ranking of six different IVIG preparations representing different manufacturing classes was identical. New generation IVIGs showed 1–2 titre steps higher anti-A titres than the older products. Haemagglutinin titres of all 48 IVIG batches analyzed were within the current Ph. Eur. specification of ≤1:64 when tested by the official pharmacopoeial method. Based on efficiency, reliability and lower costs, the direct gelcard method could be a valid alternative to the official Ph. Eur. method to serve as a limit test. However, due to the highest intermediate precision, the official Ph. Eur. method seems to be most suitable to compare haemagglutinin titres of different IVIG products.     

Isolation and Intravenous Injection of Murine Bone Marrow Derived Monocytes

As a subtype of leukocytes and progenitors of macrophages, monocytes are involved in many important processes of organisms and are often the subject of various fields in biomedical science. The method described below is a simple and effective way to isolate murine monocytes from heterogeneous bone marrow.Bone marrow from the femur and tibia of Balb/c mice is harvested by flushing with phosphate buffered saline (PBS). Cell suspension is supplemented with macrophage-colony stimulating factor (M-CSF) and cultured on ultra-low attachment surfaces to avoid adhesion-triggered differentiation of monocytes. The properties and differentiation of monocytes are characterized at various intervals. Fluorescence activated cell sorting (FACS), with markers like CD11b, CD115, and F4/80, is used for phenotyping. At the end of cultivation, the suspension consists of 45%± 12% monocytes. By removing adhesive macrophages, the purity can be raised up to 86%± 6%. After the isolation, monocytes can be utilized in various ways, and one of the most effective and common methods for in vivo delivery is intravenous tail vein injection. This technique of isolation and application is important for mouse model studies, especially in the fields of inflammation or immunology. Monocytes can also be used therapeutically in mouse disease models.