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C-terminal domain of Chk1 regulates its subcellular location and kinase activity for DNA repair

Chinese Science Bulletin 2011年第30期56卷 3138-3147页

作者： ning lifeng WANG HuiPing SAN Dan SANG JianLi Key Laboratory for Cell Proliferation and Regulation Biology of Ministry of Education College of Life Science Beijing Normal University Beijing 100875 China Department of Reproduction Immunology National Research Institute for Family Planning Beifing 100081 China

Effector kinase Chk1 is an evolutionarily conserved protein kinase. It is a key mediator linking the mechanisms that monitor DNA integrity to components of the cell cycle engine. In this study, recombinant vectors pEGFP-C1-Chk1/C 288/C 334/C 368 were constructed and transfected into HeLa cells to study the effect of the Chk1 regulatory domain on the regulation of subcellular Chk1 location in response to DNA damage. We found that DNA damage-induced nuclear accumulation is regulated by 34 amino acids (334–368) in the C-terminal regulatory domain. Recombinant vectors pXJ41-Chk1/C 288/C 334/C 368 were co-transfected with reporter plasmid pEGFP-N2 into HeLa cells to study the repair abilities of the different human Chk1 truncation mutants. In addition, recombinant vectors were transfected into HeLa cells to study the effects of the different truncation mutants on the cell cycle. Furthermore, to study the kinase activity of the different truncation mutants, Ser216 phosphorylation of Cdc25C was studied by Western blot analysis. We found that the enzymatic activity of C 368, missing the 108 C-terminal amino acids (368–476), was higher than that of full-length Chk1, and C 368 delayed the cell cycle progression. The enzymatic activity of C 334, missing the 142 C-terminal amino acids (334–476), was equivalent to that of full-length Chk1. C 288, missing the 188 C-terminal amino acids (288–476), had almost no enzymatic activity, suggesting that the regulatory domain contains both inhibitory and regulatory elements. This study provides useful information for further research on Chk1 function.

关键词：亚细胞定位激酶活性 DNA修复 HeLa细胞 DNA损伤细胞周期重组载体 blot分析

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Subcellular localization of PS1 based on PS1/GFP fusing protein

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Frontiers in Biology 2008年第1期3卷 13-18页

作者： Tie LI Jiahui LI lifeng ning Jianli SANG Institute of Cell Biology College of Life SciencesBeijing Normal UniversityBeijng 100875China Institute of Laboratory Animal Science Chinese Academy of Medical Sciences&Peking Union Medical CollegeBeijing 100021China

Mutations in presenilin 1(PS1)gene are closely associated with the early onset of familial Alzheimer’s disease(EOFAD).The fusion genes,GFP-PS1(recombinant plasmid pEGFP-C1-PS1)and PS1-GFP(recombinant plasmid pEGFP-N2-PS1)were constructed to study the subcellular localization of PS1 *** plasmids were transiently transfected into two cell lines,HEK293 and CHO,respectively,using the green fluorescence from GFP(green fluorescence protein)as the PS1 localization ***,we observed green fluorescence with a SPOT II(Olympus,BH2)and CONFOCAL microscope(Olympus,FV300)under 488 *** results show that PS1 located on the nuclear envelope.A few can be found on the cellular membrane and in the cytosol in a non-homogeneous distribution.

关键词： presenilin1 GFP fusing protein cellular distribution

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Effects of PP4 suppression on the proliferation of MCF7 cells

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Chinese Science Bulletin 2006年第19期51卷 2335-2341页

作者： ning lifeng LONG Zhitao HUANG Xiuqing SUN Lingling SANG Jianli The Key Laboratory for Cell Proliferation and Regulation Biology of Ministry of Education College of Life Science Beijing Normal University Beijing 100875

PP4, one of the few protein phosphata- ses associated with centrosome in cells of many species such as Drosophila, C. elegans and mam- mals, plays an essential role in the regulation of cen- trosome functions in Drosophila and C. elegans. In order to explore the role of PP4 in mammalian cells, full-length PP4 gene was obtained by RT-PCR from MCF7 cell total RNA and inserted into eukaryotic expression vector pEGFP-C1. The resultant con- struct pEGFP-C1-PP4 was transfected into MCF7 cells and immunostaining was carried out to confirm the centrosome localization of PP4. Then we re- versely subcloned a non-conserved domain of PP4 into pXJ41 to construct an anti-sense vector pXJ41- as-PP4. By transfecting pXJ41-as-PP4 into MCF7 cells and screening with G418, we obtained a stable cell line in which PP4 expression was stably sup- pressed. The cell line was analyzed on cell mor- phology, cytoskeleton structure, growth characteris- tics and the mitosis process. It was found that the proliferation rate decreased and serum-dependence increased in PP4-suppressed cells. Furthermore, flow cytometry and mitotic index analysis showed that G2/M transition was prolonged. PP4 suppression resulted in abnormal interphase microtubule, forma- tion of multipolar spindles and an increase in per- centage of multinuclear cells. These results sug- gested that PP4 is required for centrosome function in mammalian cells.

关键词：细胞周期 MCF7 PP4 蛋白质磷酸酶微管中心体有丝分裂反义RNA

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Potential existence of two independent centrosome-targeting domains in PP4

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Chinese Science Bulletin 2007年第21期52卷 2930-2937页

作者： HUANG XiuQing ning lifeng LONG ZhiTao SUN LingLing ZHANG Hui LIAN MuLan SANG JianLi Key Laboratory for Cell Proliferation and Regulation Biology of Ministry of Education College of Life Science Beijing NormalUniversity Beijing 100875 China

Protein phosphatase 4 (PP4) is an important member in the PPP family of protein Ser/Thr phosphatases. It has been proven to regulate a variety of cellular processes such as centrosome maturation, micro- tubule nucleation, splicesome assembly, and JNK pathway activation. Compared to the crystallized and structurally well defined phosphatase PP1 and PP2B, little is known about the structure of PP4. Besides the conserved motifs characteristic of the PPP family, no information is available on the other domains of PP4. PP4 is reported to localize to the centrosome in many species such as Drosophila, Caenor- habditis elegans and mammalian cells, which suggests a conserved role of PP4 in the regulation of centrosome function. Unlike several other centrosomal proteins, no sequence has been identified for PP4 that can target it to specific centrosomal localization. In this study, we used a combination of PCR mutagenesis and transient expression of GFP-tagged proteins in mammalian cells, and identified two PP4 centrosome-targeting domains of 68―136 and 134―220 aa. These two domains may be associated for appropriate localization to the centrosome. The findings are useful for further elucidating the func- tion of the domains and other structural characteristics of PP4.

关键词： PP4 细胞中心体局部化蛋白磷酸酶

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雌三醇的共晶研究

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医药导报 2022年第5期41卷 630-636页

作者：徐娟宁丽峰国家卫生健康委科学技术研究所生殖健康工程技术中心北京100081

目的探讨雌三醇的共晶制备以及结构表征、性能。方法利用球磨法和混悬法筛选制备雌三醇共晶,并采用粉末X射线衍射分析法、热重-差示扫描量热联用法、红外光谱法、核磁共振氢谱法等晶型结构确证方法对共晶结构进行表征,并进一步检测其稳... 详细信息

目的探讨雌三醇的共晶制备以及结构表征、性能。方法利用球磨法和混悬法筛选制备雌三醇共晶,并采用粉末X射线衍射分析法、热重-差示扫描量热联用法、红外光谱法、核磁共振氢谱法等晶型结构确证方法对共晶结构进行表征,并进一步检测其稳定性及水溶性。结果获得雌三醇-脲共晶、雌三醇-乙酰胺共晶、雌三醇-哌嗪共晶、雌三醇-异烟碱共晶、雌三醇-萘共晶、雌三醇-蒽共晶、雌三醇-菲共晶等7种共晶,并利用多种分析方法进行表征。上述共晶具有良好的物理稳定性,且具有优于雌三醇的水溶性。结论通过对雌三醇共晶的系统研究,阐明了其多种共晶样品的制备方法、表征方法,评价了其稳定性、水溶性,为雌三醇共晶药物的选择提供了科学基础数据。

关键词：雌三醇共晶稳定性水溶性

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重症监护室中心静脉导管相关性感染的预防及护理

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中国继续医学教育 2017年第15期9卷 208-209页

作者：宁利峰内蒙古自治区第四医院重症医学科内蒙古呼和浩特010010

目的对重症监护室中心静脉导管相关性感染的预防及护理进行探究分析。方法将选取2015年2月—2016年2月本院重症监护室内收治的中心静脉置管患者60例作为本次研究对象。采用简单随机分组将其分为两组,接受常规护理的30例患者为对照组,在... 详细信息

目的对重症监护室中心静脉导管相关性感染的预防及护理进行探究分析。方法将选取2015年2月—2016年2月本院重症监护室内收治的中心静脉置管患者60例作为本次研究对象。采用简单随机分组将其分为两组,接受常规护理的30例患者为对照组,在其基础上,开展综合护理的30例患者为观察组,对两组患者中心静脉导管相关性感染发生概率、ICU住院天数进行比较。结果观察组患者的中心静脉导管总感染概率低于对照组,其ICU住院天数为(6.93±1.35)天少于对照组,差异具有统计学意义(P<0.05)。结论在重症监护中实施综合护理可明显降低中心静脉导管相关性感染发生的概率。

关键词：重症监护室中心静脉导管相关性感染综合护理

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Impacts of the annual cycle of background SST in the tropical Pacific on the phase and amplitude of ENSO

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Atmospheric and Oceanic Science Letters 2025年第1期18卷 12-17页

作者： Song Jiang Congwen Zhu ning Jiang State Key Laboratory of Severe Weather(LASW) Chinese Academy of Meteorological SciencesBeijingChina College of Earth and Planetary Sciences University of Chinese Academy of SciencesBeijingChina

The dominant annual cycle of sea surface temperature(SST)in the tropical Pacific exhibits an antisymmetric mode,which explains 83.4%total variance,and serves as a background of El Niño-Southern Oscillation(ENSO).However,there is no consensus yet on its anomalous impacts on the phase and amplitude of *** on data during 1982-2022,results show that anomalies of the antisymmetric mode can affect the evolution of ENSO on the interannual scale via Bjerknes feedback,in which the positive(negative)phase of the antisymmetric mode can strengthen El Niño(La Niña)in boreal winter via an earlier(delayed)seasonal cycle transition and larger(smaller)annual *** magnitude of the SST anomalies in the equatorial eastern Pacific can reach more than±0.3◦C,regulated by the changes in the antisymmetric mode based on random sensitivity *** reveal the spatial pattern of the annual cycle associated with the seasonal phase-locking of ENSO evolution and provide new insight into the impact of the annual cycle of background SST on ENSO,which possibly carries important implications for forecasting ENSO.

关键词： Annual cycle SST anomaly Antisymmetric mode ENSO

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The pivotal role of microglia in injury and the prognosis of subarachnoid hemorrhage

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Neural Regeneration Research 2025年第7期20卷 1829-1848页

作者： Wenjing ning Shi Lv Qian Wang Yuzhen Xu Department of Rehabilitation The Second Affiliated Hospital of Shandong First Medical UniversityTaianShandong ProvinceChina Department of Central Laboratory The Affiliated Taian City Central Hospital of Qingdao UniversityTaianShandong ProvinceChina

Subarachnoid hemorrhage leads to a series of pathological changes,including vascular spasm,cellular apoptosis,blood–brain barrier damage,cerebral edema,and white matter ***,which are the key immune cells in the central nervous system,maintain homeostasis in the neural environment,support neurons,mediate apoptosis,participate in immune regulation,and have neuroprotective *** evidence has shown that microglia play a pivotal role in the pathogenesis of subarachnoid hemorrhage and affect the process of injury and the prognosis of subarachnoid ***,microglia play certain neuroprotective roles in the recovery phase of subarachnoid *** approaches aimed at modulating microglia function are believed to attenuate subarachnoid hemorrhage *** provides new targets and ideas for the treatment of subarachnoid ***,an in-depth and comprehensive summary of the role of microglia after subarachnoid hemorrhage is still *** review describes the activation of microglia after subarachnoid hemorrhage and their roles in the pathological processes of vasospasm,neuroinflammation,neuronal apoptosis,blood–brain barrier disruption,cerebral edema,and cerebral white matter *** also discusses the neuroprotective roles of microglia during recovery from subarachnoid hemorrhage and therapeutic advances aimed at modulating microglial function after subarachnoid ***,microglia in subarachnoid hemorrhage are targeted with TLR inhibitors,nuclear factor-κB and STAT3 pathway inhibitors,glycine/tyrosine kinases,NLRP3 signaling pathway inhibitors,Gasdermin D inhibitors,vincristine receptorαreceptor agonists,ferroptosis inhibitors,genetic modification techniques,stem cell therapies,and traditional Chinese ***,most of these are still being evaluated at the laboratory *** clinical studies and data on subarachnoid hemorrhage are required to improve the treatment of subarachnoid hemor

关键词： apoptosis blood–brain barrier brain edema microglia neuroinflammation neuron neuroprotection subarachnoid hemorrhage vasoconstriction white matter injury

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炔雌醇共晶药动学研究

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医药导报 2022年第3期41卷 283-286页

作者：宁丽峰杜荣凯张雷徐娟国家卫生健康委科学技术研究所北京100081 华南理工大学化学与化工学院广州510641

目的研究炔雌醇4种共晶(炔雌醇-哌嗪、炔雌醇-烟酰胺、炔雌醇-川芎嗪、炔雌醇-咪唑)在大鼠体内的吸收与代谢,探讨炔雌醇共晶药动学研究方法。方法雌性SD大鼠口服给药,剂量为5.0 mg·kg^(-1)(炔雌醇相当量),于规定时间分批取血。结果炔... 详细信息

目的研究炔雌醇4种共晶(炔雌醇-哌嗪、炔雌醇-烟酰胺、炔雌醇-川芎嗪、炔雌醇-咪唑)在大鼠体内的吸收与代谢,探讨炔雌醇共晶药动学研究方法。方法雌性SD大鼠口服给药,剂量为5.0 mg·kg^(-1)(炔雌醇相当量),于规定时间分批取血。结果炔雌醇达峰时间1 h,于24 h内代谢完毕;4种共晶达峰时间约为1.5 h,于48 h内代谢完毕;炔雌醇血药浓度-时间曲线下面积(AUC)为(25.1±2.8)h·ng·mL^(-1),炔雌醇-哌嗪共晶AUC(28.3±1.5)h·ng·mL^(-1),炔雌醇-烟酰胺共晶AUC(26.8±2.2)h·ng·mL^(-1),炔雌醇-川芎嗪共晶AUC(24.2±4.4)h·ng·mL^(-1),炔雌醇-咪唑共晶AUC(23.6±2.5)h·ng·mL^(-1)。结论炔雌醇形成共晶后,药物释放时间延长;通过制备炔雌醇-哌嗪共晶和炔雌醇-烟酰胺共晶可提高炔雌醇在大鼠体内的生物利用度。

关键词：炔雌醇共晶药动学血药浓度-时间曲线下面积

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Coordinated Control Strategy of New Energy Power Generation System with Hybrid Energy Storage Unit

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Energy Engineering 2025年第1期122卷 167-184页

作者： Yun Zhang ZifenHan Biao Tian ning Chen Yi Fan School of Mechano-Electronic Engineering Xidian UniversityXi’an710071China Electric Power Control Center State Grid Gansu Electric Power CompanyLanzhou730030China Electric Power Control Center State Grid Gansu Electric Power Company Zhangye Power Supply BranchZhangye734000China

The new energy power generation is becoming increasingly important in the power *** as photovoltaic power generation has become a research hotspot,however,due to the characteristics of light radiation changes,photovoltaic power generation is unstable and random,resulting in a low utilization rate and directly affecting the stability of the power *** solve this problem,this paper proposes a coordinated control strategy for a newenergy power generation system with a hybrid energy storage unit based on the lithium iron phosphate-supercapacitor hybrid energy storage ***,the variational mode decomposition algorithm is used to separate the high and low frequencies of the power signal,which is conducive to the rapid and accurate suppression of the power fluctuation of the energy storage ***,the fuzzy control algorithm is introduced to balance the power between energy *** this paper,the actual data is used for simulation,and the simulation results show that the strategy realizes the effective suppression of the bus voltage fluctuation and the accurate control of the internal state of the energy storage unit,effectively avoiding problems such as overshoot and over-discharge,and can significantly improve the stability of the photovoltaic power generation systemand the stability of the Direct Current *** is of great significance to promote the development of collaborative control technology for photovoltaic hybrid energy storage units.

关键词： Photovoltaic power suppression hybrid energy storage unit variationalmodal decomposition fuzzy control power distribution control

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