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Prokaryotic expression of recombinant human p75NTR-Fc fusion protein and its effect on the neurite outgrowth of dorsal root ganglia neuron

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Journal of Medical Colleges of PLA(China) 2009年第1期24卷 1-9页

作者： Zhu Feng Wang Yongtang Lu Xiumin Zeng Lin Wu Yamin College of Chemical and Biological Engineering Chongqing Institute of Technology Chongqing 400050 China state key laboratory of Trauma Burns and Combined Injury Institute of Surgery Research Daping Hospital Third Military Medical University Chongqing 400042 China

Objective： To clone, express, and identify the extracellular domain gene of human p75 neurotrophin receptor with IgG-Fe （hp75NTR-Fc） in prokaryotic expression system, and investigate the effect of the recombinant protein on dorsal root ganglia （DRG） neuron neurites. Methods： The hp75NTR-Fc coding sequence was amplified from pcDNA-hp75NTR-Fc by polymerase chain reaction （PCR） and subcloned into vector pET30a （＋）, in which hp75NTR-Fc expression was controlled under the T7 promoter. The recombinant vectors were amplified in E. coli DH5α and identified by PCR, enzyme digestion and sequencing, and then transformed into E. coli BL21 （DE3）. The expression product was analyzed with SDS-PAGE and Western blot. Then after the recombinant protein purified with Protein A affinity chromatograph, and renaturated with dialysis, respectively, the effect of the recombinant protein on DRG neuron neuritis was further investigated. Results： The results of PCR, enzyme digestion, and sequencing demonstrated the success of inserting the hp75NTR-Fc fragment into vector pET30a （＋）. SDS-PAGE and Western blot showed a positive protein band with molecular weight about 50 kD in the expression product, which is accordant with the interest protein, and this band could be specifically recognized by rabbit anti-NGFRp75 antibody. The purified infusion protein following dialysis could promote neurite outgrowth of DRG neurons cultured with myelin-associated glycoprotein （MAG）. Conclusion： The hp75NTR-Fc coding sequence was subcloned into the expression vector pET30a （＋） correctly and expressed successfully in the prokaryotie expression system. The infusion protein could promote neurite outgrowth of DRG neurons cultured with MAG.

关键词： p75NTR Vector construction Prokaryotic expression Dorsal root ganglia neuron Neurite outgrowth

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Expression of NF-κB in Schwann cells and its effect on motor neuron apoptosis in spinal cord following sciatic nerves injury in rats

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Journal of Medical Colleges of PLA(China) 2007年第2期22卷 92-96页

作者：王永堂鲁秀敏余瑛杨艳红高洁 state key laboratory of Trauma Burns and Combined Injury Institute of Surgery ResearchDaping HospitalThird Military Medical University College of Bioengineering Chongqing Institute of Technology

Objective：To explore the expression of nuclear factor-kappa B （NF-kB） in Schwann cells （SCs） and its effect on motor neuron apoptosis in spinal cord following sciatic nerves injury in adult rats. Methods： Thirty-six adult Sprague-Dawley （SD） rats were divided randomly into normal control group （n=6）, and sciatic nerves crushing group （n= 30）, and the later was further equally randomized into 5 subgroups： 1, 3, 7, 14, and 21 d post-injury groups. The expression of NF-kB of normal and injured nerves were examined by immunohistochemistry staining, and the apoptosis of motor neurons in spinal cord of lumbar 4 to lumbar 6 （L4-L6） was investigated by terminal deoxynucleotidyl transferase mediated dUTP-biotin nick end labeling （TUNEL） assay. Both were *** by image analysis. Results： In crushing group, except 21 d post-injury group, the expression of NF-kB was markedly higher than that in the normal control group （P〈0.05, P〈0. 01）. At 1 d after sciatic nerves crushing, the expression of NF-kB was obviously up-regulated, reached peak at 3 d, and recovered at 21 d. The same trend was observed in the time-course on motor neuron apoptosis after sciatic nerves injury. Correlation analyses revealed that motor neuron apoptosis was significantly and positively correlated with the expression of NF-kB following sciatic nerves injury （r= 0. 976 0, P〈0. 01）. Conclusion： After injury of sciatic nerves, the presence and up-regulation of NF-kB in SCs may be involved in motor neuron apoptosis in L4-L6 spinal cord.

关键词： sciatic nerves injury nuclear factor-kappa B motor neuron apoptosis spinal cord

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模拟水体湍流环境下目标激光点云数据的三维重建与分析

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光电工程 2023年第6期50卷 56-65页

作者：王明军彭月刘燕荣李勇俊李乐西安理工大学自动化与信息工程学院陕西西安 710048 西安市无线光通信和网络研究重点实验室陕西西安 710048 陕西省智能协同网络军民共建重点实验室陕西西安 710126 散射辐射全国重点实验室北京 100854 天津津航技术物理研究所天津 300308 西安理工大学自动化与信息工程学院陕西西安 710048

水下目标三维点云重建在水底勘探和水下无人航行器避障等领域有着重要的应用.本文通过实验室模拟温度和盐度水体湍流,开展了水下目标探测实验,研究了温度差和盐度差对水下潜艇、滑翔机和锚雷的激光点云三维重建效果的影响,并对重建后的... 详细信息

水下目标三维点云重建在水底勘探和水下无人航行器避障等领域有着重要的应用.本文通过实验室模拟温度和盐度水体湍流,开展了水下目标探测实验,研究了温度差和盐度差对水下潜艇、滑翔机和锚雷的激光点云三维重建效果的影响,并对重建后的三维点云数据进行误差分析.结果表明:在水下目标三维点云重建过程中,随着水体温度湍流和盐度湍流强度的增强,重建点云数据的有效点数明显下降,均值误差明显上升.研究结果对水下三维点云重建系统的研制具有一定参考价值.

关键词：水下探测水体湍流模拟三维点云重建误差分析

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Continuous purification and culture of rat type 1 and type 2 alveolar epithelial cells by magnetic cell sorting

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Chinese Journal of Traumatology 2022年第3期25卷 138-144页

作者： Di Liu Jian-Hui Sun Hua-Cai Zhang Jian-Xin Jiang Ling Zeng state key laboratory of Trauma Bums and Combined InjuriesDoping HospitalArmy Medical UniversityChongqing400042China Medical Center of Trauma and War Injury Daping HospitalArmy Medical UniversityChongqing400042China

Purpose:The incidence of acute lung injury(ALI)in severe trauma patients is 48%and the mortality rate following acute respiratory distress syndrome evolved from ALI is up to 68.5%.Alveolar epithelial type 1 cells(AEC1 s)and type 2 cells(AEC2s)are the key cells in the repair of injured lungs as well as fetal lung ***,the purification and culture of AECls and AEC2s play an important role in the research of repair and regeneration of lung ***:Sprague-Dawley rats(3-4 weeks,120-150 g)were purchased for *** and DNase I were jointly used to digest lung tissue to obtain a single-cell suspension of whole lung cells,and then magnetic bead cell sorting was performed to isolate Tla positive cells as AECls from the single-cell suspension by using polyclonal rabbit anti-Tla(a specific AECls membrane protein)antibodies combined with anti-rabbit IgG ***,alveolar epithelial cell membrane marker protein EpCAM was designed as a key label to sort AEC2s from the remaining Tla-neg cells by another positive immunomagnetic selection using monoclonal mouse anti-EpCAM antibodies and anti-mouse IgG *** purity was identified by immunofluorescence staining and flow ***沾••The purity of AECls and AEC2s was 88.3%±3.8%and 92.6%±2.7%,*** cell growth was observed as follows:AECls stretched within the 12-16 h,but the cells proliferated slowly;while AEC2s began to stretch after 24 h and proliferated rapidly from the 2nd day and began to differentiate after 3 ***:AECls and AEC2s sorted by this method have high purity and good ***,our method provides a new approach for the isolation and culture of AECls and AEC2s as well as a new strategy for the research of lung repair and regeneration.

关键词： Alveolar epithelial cells type 1 Alveolar epithelial cells type 2 Three-dimensional culture Magnetic activated cell sorting

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The injury progression of T lymphocytes in a mouse model with subcutaneous injection of a high dose of sulfur mustard

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Journal of Medical Colleges of PLA(China) 2014年第3期29卷 162-172页

作者： Yi-Zhou Mei Xiao-Rui Zhang Ning Jiang Jun-Ping Cheng Feng Liu Pan Zheng Wen-Xia Zhou Yong-Xiang Zhang state key laboratory of Toxicology and Medical Countermeasures Beijing Institute of Pharmacology and Toxicology

Background: In clinical studies, the findings on sulfur mustard(SM) toxicity for CD3+CD4+ and CD3+CD8+ T lymphocyte subsets are contradictory. In animal experiments, the effect of SM on the T cell number and proliferation is incompatible and is even the opposite of the results in human studies. In this study, we observed the dynamic changes of T lymphocytes in the first week in a high-dose SM-induced ***: Mice were exposed to SM by subcutaneous injection(20 mg/kg) and were sacrificed 4 h, 24 h, 72 h and 168 h later. Spleen T lymphocyte proliferation was evaluated by 3H-Td R. Flow cytometric analysis was used to observe the percentage of CD3+CD4+ and CD3+CD8+ T lymphocyte subsets. The IL-1e assayed using the Luminex method. DNA damage in bone marrow ceβ, IL-6, IL-10 and TNF-lls was observed with α levels in plasma werthe single cell gel electrophoresis technique(SCGE).Results: SM continuously inhibited the proliferation of lymphocytes for 7 days, and there was a significant rebound of Con A-induced T lymphocyte proliferation only at 24 h. The percentage of CD3+CD4+ and CD3+CD8+ lymphocytes was upregulated, which was accompanied by increased IL-1β and TNF-creased in the PG group at 4 h. The peak of lymphocytic apoptα and decreased IL-10. The IL-6 level was gradually deosis and DNA damage occurred at 24 h and 72 h, respectively. Conclusion: Our results show that SM significantly inhibited T lymphocyte proliferation as well as induced CD3+CD4+ and CD3+CD8+ upregulation. SM intoxication also significantly increased the levels of pro-inflammatory cytokines(IL-1β, IL-6 and TNF-α) and inhibited the level of anti-inflammatory cytokine IL-10. Our results may partly be due to the significant SM induced significant apoptosis and necrosis of lymphocytes as well as DNA damage of bone marrow cells. The results provided a favorable evaluation of SM immune toxicity in an animal model.

关键词： Sulfur Mustard T lymphocyte Apoptosis Cytokine DNA damage

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A metabolic acidity-activatable calcium phosphate probe with fluorescence signal amplification capabilities for non-invasive imaging of tumor malignancy

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Science Bulletin 2022年第3期67卷 288-298页

作者： Qian Jia Ruili Zhang Yongdong Wang Haohao Yan Zheng Li Yanbin Feng Yu Ji Zuo Yang Yang Yang Kanyi Pu Zhongliang Wang laboratory of Molecular Imaging and Translational Medicine(MITM) Engineering Research Center of Molecular&NeuroimagingMinistry of EducationSchool of Life Science and TechnologyXidian UniversityXi’an 710126China Academy of Advanced Interdisciplinary Research Xidian UniversityXi’an 710071China Department of Thoracic Surgery Shanghai Pulmonary HospitalSchool of MedicineTongji UniversityShanghai 200433China School of Materials Science and Engineering Tongji UniversityShanghai 201804China School of Chemical and Biomedical Engineering Nanyang Technological UniversitySingapore 637457Singapore

Dysregulated energy metabolism has recently been recognized as an emerging hallmark of *** cells,which are characterized by abnormal glycolysis,exhibit a lower extracellular pH(6.5–7.0)than nor-mal tissues(7.2–7.4),providing a promising target for tumor-specific imaging and ***,most pH-sensitive materials are unable to distinguish such a subtle pH difference owing to their wide and continuous pH-responsive *** this study,we developed an efficient strategy for the fabrication of a tumor metabolic acidity-activatable calcium phosphate(CaP)fluorescent probe(termed MACaP9).Unlike traditional CaP-based biomedical nanomaterials,which only work within more acidic organelles,such as endosomes and lysosomes(pH 4.0–6.0),MACaP9 could not only specifically respond to the tumor extra-cellular pH but also rapidly convert pH variations into a distinct fluorescence signal to visually dis-tinguish tumor from normal *** superior sensitivity and specificity of MACaP9 enabled high-contrast visualization of a broad range of tumors,as well as small tumor lesions.

关键词： Calcium phosphate Activatable probe Cancer imaging Tumor microenvironment Tumor metabolism

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