Soluble Prokaryotic Overexpression and Purification of Human GM-CSF Using the Protein Disulfide Isomerase b′a′ Domain

Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a member of the colony-stimulating factor (CSF) family, which functions to enhance the proliferation and differentiation of hematopoietic stem cells and other hematopoietic lineages such as neutrophils, dendritic cells, or macrophages. These proteins have thus generated considerable interest in clinical therapy research. A current obstacle to the prokaryotic production of human GM-CSF (hGM-CSF) is its low solubility when overexpressed and subsequent complex refolding processes. In our present study, the solubility of hGM-CSF was examined when combined with three N-terminal fusion tags in five E. coli strains at three different expression temperatures. In the five E. coli strains BL21 (DE3), ClearColi BL21 (DE3), LOBSTR, SHuffle T7 and Origami2 (DE3), the hexahistidine-tagged hGM-CSF showed the best expression but was insoluble in all cases at each examined temperature. Tagging with the maltose-binding protein (MBP) and the b′a′ domain of protein disulfide isomerase (PDIb′a′) greatly improved the soluble overexpression of hGM-CSF at 30 °C and 18 °C. The solubility was not improved using the Origami2 (DE3) and SHuffle T7 strains that have been engineered for disulfide bond formation. Two conventional chromatographic steps were used to purify hGM-CSF from the overexpressed PDIb′a′-hGM-CSF produced in ClearColi BL21 (DE3). In the experiment, 0.65 mg of hGM-CSF was isolated from a 0.5 L flask culture of these E. coli and showed a 98% purity by SDS-PAGE analysis and silver staining. The bioactivity of this purified hGM-CSF was measured at an EC₅₀ of 16.4 ± 2 pM by a CCK8 assay in TF-1 human erythroleukemia cells.     

Importance of Low-Temperature Melt-Mixing on the Construction of Stereocomplex Crystallites with Superior Nucleation Efficiency in Asymmetric Poly(l-lactide)/Poly(d-lactide) Blends

The nucleation efficiency (NE) of stereocomplex crystallites (SCs) formed in asymmetric poly(L-lactide)/poly(D-lactide) (PLLA/PDLA) blends is generally unsatisfactory because the competition between stereocomplexation and chain mixing involved in the melt-mixing process can cause low formation efficiency and even severe aggregation of SCs. Herein, it is attempted to achieve high-efficient formation of finely dispersed SCs particles by designing a unique melt-mixing procedure, where the mixing of PLLA with 0.75 wt% PDLA is first performed at elevated temperatures (far above the melting temperature of SCs) to allow the homogeneous mixing of PLLA/PDLA chains and then at a low temperature (slightly above that of homocrystallites) to permit the full stereocomplexation of the premixed chains. It is found that the SCs formed in the blends exhibit unexpectedly low NEs (e.g., 54.5%), much inferior to that (73.6%) in the counterpart without undergoing premixing. This is because the introduction of premixing leads to a remarkable deterioration in the amount of SCs particles formed, despite decreased particle size, highlighting that the direct mixing at low temperatures of 170–180 °C (about 20–30 °C lower than that used in common melt-processing of PLA) is more effective for the construction of SCs with superior NE. The mechanisms for these striking findings are discussed.     

含硫氰酸盐、硫化物固体废物中总氰化合物测定方法

实验建立了含硫氰酸盐、硫化物固体废物中总氰化合物的测定方法,采用硝酸银滴定法、异烟酸-吡唑啉酮分光光度法分别测定总氰化合物含量高、低的样品。通过条件实验优化了前处理方式、称样量和干扰去除方法。该方法测定结果的相对标准偏差为0.43%～1.33%,加入标准物质回收率为91.5%～101.5%,精密度和准确度良好,符合测定要求。     

氧化铝超浓相输送系统计量新装置的研究开发

本文介绍了我国首例铝电解大组氧化铝超浓相输送管路上安装电子秤进行氧化铝静态称量连续下料自动计量新方法、新装置及其使用精度的有关基本情况。     

准贝氏体铸钢正火热处理工艺的研究

研究了准贝氏体铸钢不同奥氏体温度正火对其组织与力学性能的影响 ,结果表明 ,随奥氏体化温度的提高 ,准贝氏体铸钢的强韧性改善 ,在 10 4 0℃正火 ,准贝氏体铸钢具有最佳强韧性配合。分析了高温奥氏体化正火强韧性提高的原因。     

Effect of lamellar spacing on microstructural instability and creep behavior of a lamellar TiAl alloy

Finer lamellar spacing in the lamellar structure of a Ti–45Al–2Nb–2Mn + 0.8 vol.%TiB₂ (45XD) alloy does improve the primary creep resistance. However, the unstable nature of the fine plate contributes largely to the degradation of the lamellar structure and a rapid increase in the tertiary creep rate, indicating that a fine lamellar structure has a detrimental effect on the long-term creep.     

9SiCr钢中的筐篮编结状马氏体及其性能

将9SiCr钢的粒状原始组织改变为片状,再经830℃淬火后,其马氏体形态为筐篮编结状。而传统工艺所获得的马氏体形态为针状.这种筐篮编结状马氏体是9SiCr钢中一种崭新的组织形态,其精细结构为平行成束的板条。亚结构为高密度缠结位错.它有良好的综合机械性能,强韧性高,耐磨性高。     

Curing behavior and structure of an epoxy/clay nanocomposite system

The curing behavior of an epoxy/clay nanocomposite system composed of a bifunctional epoxy resin with an aromatic amine curing agent and an organically modified clay was investigated. Differential scanning calorimetry (DSC) was used to investigate the curing behavior of the epoxy/clay nanocomposite system. The curing rate of the nanocomposite system increased with increasing clay content. A kinetic equation, considering an autocatalytic reaction mechanism, could describe fairly well the curing behavior of the epoxy/clay nanocomposite system. The reaction kinetic parameters of the kinetic equation were determined by fitting DSC conversion data to the kinetic equation, using a nonlinear numerical method. Dynamic mechanical analysis was used to investigate the thermomechanical properties of the epoxy/clay nanocomposite system. The glass transition temperature of the epoxy/clay nanocomposite system increased slightly with increasing clay content. The structure of the nanocomposite system was characterized by X‐ray diffraction analysis and transmission electron microscope imaging. The formation of intercalated structures was observed dominantly in the epoxy/clay nanocomposites, together with some exfoliated structures. POLYM. ENG. SCI., 46:1318–1325, 2006. © 2006 Society of Plastics Engineers     

An experimental investigation of mixing of carbon nanotube/polymer composite in a batch-type screw mixer

We develop a miniaturized batch-type screw mixer (BSM) for uniform mixing of polymer resin and nanoparticles, based on the stretching of material elements. This stretching is induced by the combination of recirculating cross-sectional flows in deep channels of the screw and high shear stress developed at flight regions. The BSM is used to produce a polymer nano-composite composed of multi-walled carbon nanotubes and polydimethylsiloxane resin. The mixing performance of the BSM is characterized quantitatively by estimating two different types of mixing efficiencies (i.e., dispersive mixing and distributive mixing) via transmitted light microscope images. The developed BSM highly improves the mixing performance rather than that of a conventional ultrasonic mixing device.     

类球形低硫磷酸铁的制备技术研究

磷酸铁（FePO₄）是锂电池正极材料磷酸铁锂（LiFePO₄）的核心前驱体,FePO₄形貌及硫含量对合成的LiFePO₄材料性能有重要影响。为得到类球形低硫FePO₄产品,在传统液相沉淀法技术基础上做了改进优化,添加十六烷基三甲基溴化铵（CTAB）作为形貌助剂提高产品球形度,添加氨水作为配体形成磷酸铁铵配合物改善结晶过程,降低产品硫含量。结果表明：所制备的FePO₄产品硫质量分数低,达到2.6×10 ^-5,形貌为均一的微米类球形颗粒,D₅₀=11.4 μm,振实密度达到1.22 g/cm ³,有望成为制备高压实密度LiFePO₄材料的核心前驱体。