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.     

An Engineered Biliverdin-Compatible Cyanobacteriochrome Enables a Unique Ultrafast Reversible Photoswitching Pathway

Cyanobacteriochromes (CBCRs) are promising optogenetic tools for their diverse absorption properties with a single compact cofactor-binding domain. We previously uncovered the ultrafast reversible photoswitching dynamics of a red/green photoreceptor AnPixJg2, which binds phycocyanobilin (PCB) that is unavailable in mammalian cells. Biliverdin (BV) is a mammalian cofactor with a similar structure to PCB but exhibits redder absorption. To improve the AnPixJg2 feasibility in mammalian applications, AnPixJg2_BV4 with only four mutations has been engineered to incorporate BV. Herein, we implemented femtosecond transient absorption (fs-TA) and ground state femtosecond stimulated Raman spectroscopy (GS-FSRS) to uncover transient electronic dynamics on molecular time scales and key structural motions responsible for the photoconversion of AnPixJg2_BV4 with PCB (Bpcb) and BV (Bbv) cofactors in comparison with the parent AnPixJg2 (Apcb). Bpcb adopts the same photoconversion scheme as Apcb, while BV4 mutations create a less bulky environment around the cofactor D ring that promotes a faster twist. The engineered Bbv employs a reversible clockwise/counterclockwise photoswitching that requires a two-step twist on ~5 and 35 picosecond (ps) time scales. The primary forward P_fr → P_o transition displays equal amplitude weights between the two processes before reaching a conical intersection. In contrast, the primary reverse P_o → P_fr transition shows a 2:1 weight ratio of the ~35 ps over 5 ps component, implying notable changes to the D-ring-twisting pathway. Moreover, we performed pre-resonance GS-FSRS and quantum calculations to identify the Bbv vibrational marker bands at ~659,797, and 1225 cm⁻¹. These modes reveal a stronger H-bonding network around the BV cofactor A ring with BV4 mutations, corroborating the D-ring-dominant reversible photoswitching pathway in the excited state. Implementation of BV4 mutations in other PCB-binding GAF domains like AnPixJg4, AM1_1870g3, and NpF2164g5 could promote similar efficient reversible photoswitching for more directional bioimaging and optogenetic applications, and inspire other bioengineering advances.     

基于二极管-电容倍压单元的DC-DC升压变换器

文章基于二极管-电容倍压单元,提出了一种新型高增益升压变换器。该拓扑在实现高增益的同时可以避免使用极大的占空比,并且有效地降低开关管和二极管的电压应力。分析了变换器的工作原理,推导了变换器工作在连续状态（CCM）和断续状态（DCM）下的电压增益、开关器件的电压应力和电感电流纹波。CCM状态下与传统的Boost变换器相比电压增益是其（5-D）倍,电感电流纹波近似减小为原来的一半。最后通过仿真与样机实验验证了理论分析的正确性,表明提出的变换器具有良好的综合性能。     

并网变换器电流重构模型预测容错控制

当传统并网变换器发生桥臂故障时,通过连接故障相到直流侧电容中点,重构为三相四开关容错运行结构。针对三相四开关容错变换器的交流电流传感器故障问题,提出了一种基于电流重构的模型预测功率控制方法。首先,利用采样的直流电流和正常交流传感器信息,并根据变换器电压矢量与电流关系重新构造出三相电流。其次,建立容错变换器功率预测模型,利用重构电流参与容错变换器的模型预测控制,应用使代价函数取得最小值的开关矢量。最后,搭建仿真和实验平台进行验证,结果表明所提控制策略有效,输出功率稳定,实现了并网变换器在桥臂和传感器双重故障下的容错运行,提高了系统的可靠性。     

考虑城市场景需求的中压直流配电网规划选优分析方法

针对城市配电网规划中多个中压直流配电网的建设优先次序尚无量化分析方法的问题,提出一种考虑城市场景需求的中压直流配电网规划选优分析方法.从供电质量提升需求、城市场景需求和配电网经济性3个层面建立了一套城市中压直流配电网规划选优综合指标体系,在供电质量提升需求方面考虑与目标水平之间的差距,在城市场景需求方面考虑中压交流配电网的建设局限性与中压直流配电网的建设驱动力,在经济性方面关注缺电损失成本严重性.通过云模型对定性指标量化,基于德尔菲-熵权法的层次加性加权计算各配电网综合评价值,并以此排序,在年度总投资约束下,以方案总体综合评价值最大为目标实现规划对象选优,从而安排多个中压直流配电网的建设优先级.以某地区配电网为例,验证了所提方法的实用性.     

高风电渗透率下考虑电网频率支撑需求的储能系统配置方法

提出了一种高风电渗透率下考虑电网频率支撑需求的储能系统配置方法.以频率变化率和频率偏差为限制条件,建立新能源系统所能承受的最大功率增量与等效惯性常数、调差系数以及风电渗透率等已知参数的联系.通过对3阶虚拟同步机控制策略下的储能系统容量与控制参数进行量化配置,提高不同风电渗透率系统对不平衡功率的消纳能力.以储能配置在频率支撑中贡献的等效单位调节功率为参考,对不同功率增量下储能系统的频率响应贡献、调频出力占比以及输出功率特性进行刻画与分析.仿真验证了该配置方法下的储能系统可控性强,能够较为精准地提供电网所需的有功调节量,有效改善风电并网环境.     

核级FeCrAl包壳材料研究进展

摘要：日本福岛核电站泄漏事故暴露了锆合金易与水蒸气反应造成爆炸的缺陷,又将人们的视线拉回了各项性能较为均衡的FeCrAl合金。FeCrAl合金较锆合金具有更好的耐腐蚀性能,更能满足核反应堆高温水蒸气环境下的工作条件。为进一步保障核反应堆安全性,各国学者对FeCrAl合金的性能进行了大量研究。基于此,综述了FeCrAl合金腐蚀机制及其在高温蒸汽和高温水环境中的腐蚀行为;探讨了合金元素（稀土、Ti、Zr、Nb和Mo）、变形和热处理对合金力学性能的影响;总结了FeCrAl合金辐照性能和焊接性能的研究进展。最后指出了核级FeCrAl合金在研究中存在的不足及未来重点发展方向,为今后其他学者在该领域的研究提供参考。     

鞍钢鲅鱼圈4038m3高炉高比例使用中块焦炭生产实践

针对鞍钢股份有限公司鲅鱼圈钢铁分公司4038 m3高炉高比例使用中块焦炭后出现的风量萎缩、顺行变差,尤其是高炉休送风恢复过程中炉缸活跃性差、煤气流中心通道打不开、频繁崩滑料现象,提出采取合理匹配上下部调剂、组织好炉前渣铁排放、小焦布料制度以及改善休风料结构等措施.实践后,高炉实现了中块焦比为90 kg/t情况下的长期稳定顺行,休送风恢复未出现超时现象,适应了高比例使用中块焦炭的生产.     

X80管线钢埋弧焊匹配焊丝试验

通过分析高强度低合金钢焊缝熔敷金属常见显微组织对其力学性能的影响,确定X80管线钢用埋弧焊丝熔敷金属组织应以大量针状铁素体(AF)和少量粒状贝氏体(GB)的复合组织.从相变动力学原理出发,结合针状铁素体(AF)非自发形核机制和微合金组织韧化理论,选择Mn-Ni-Mo-Ti-B合金系进行X80管线钢匹配焊丝的试制.结果表明,合理选择和控制合金元素,可以获得理想的焊缝熔敷金属组织和强韧性以及低温韧性要求,试制的1号焊丝能够满足X80管线钢的使用要求.