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Expression of Human Vascular Endothelial Growth Factor (VEGF_(165)) in Pichia pastoris and Its Biological Activity

Biomedical and Environmental Sciences 2001年第4期14卷 302-311页

作者： MALI zhangzhi－qing 等 InstituteofMolecularImmunology theFirstMilitaryMedicalUniversityGuangzhou510515China StateKeyLaboratoryofMolecularVirologyandGeneticEnginering Beijing100052China

Objective To express human vascular endothelial growth factor (hVEGF-{165}) cDNA in %Pichia pastroris, %purify the expressed product and detect the biological activity of it. Methods By inserting hVEGF-{165} cDNA coding 165 amino acid residues into %Pichia pastoris% expression vector pPIC9K containing AOX1 promoter and the sequences of α secreting signal peptides, a recombinant expression plasmid pPIC9K/hVEGF-{165} was constructed and transformed to yeast host strain KM71, then multiple_copy insert transformants were screened out and cultured in flasks, and hVEGF-{165} was expressed under the induction of 1% methanol. Results SDS_PAGE showed that after being induced with 1% methanol for 4d, the expressed product existed in supernatant in the form of soluble molecule and contained 60% of total protein expressed. Western blot showed good antigenicity and specificity of expressed product. After being purified by Heparin_Sepharose CL6B affinity chromatography, the purity of expressed product reached above 90%. Biological assays proved that the expressed product could stimulate the proliferation of HUVEC. Conclusion hVEGF-{165} was successfully expressed. The study opened up a wide prospect for the application of VEGF-{165} in the prevention and treatment of ischemic heart disease and other tissue ischemic diseases such as secondary arterial occlusion in limbs.

关键词： Base Sequence Blotting Western Chromatography Affinity DNA Primers Electrophoresis Polyacrylamide Gel Endothelial Growth Factors Humans Lymphokines Pichia Plasmids Polymerase Chain Reaction Recombinant Proteins Research Support Non-U.S. Gov't Vascular Endothelial Growth Factor A Vascular Endothelial Growth Factors

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Expression of Human Vascular Endothelial Growth Factor Receptor Flt-1 Extracellular Domain 1-3 Loop cDNA in Pichia pastoris, Purification of the Expressed Product and Detection of Its Biological Activity

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Biomedical and Environmental Sciences 2001年第4期14卷 292-301页

作者： MALI zhangzhi－qing 等 InstituteofMolecularImmunology ThefirstMilitaryMedicalUniversityGuangzhou510515China StateKeyLaboratoryofMolecularVirologyandGeneticEngineering Beijing100052China

Objective To express human vascular endothelial growth factor receptor Flt-1 extracellular domain 1-3 loop cDNA in Pichia. pastroris, and to purify the expressed product and detect its biological activity. Methods By inserting human Flt-1 (1-3 loop) cDNA coding 316 amino acid residues into Pichia pastoris expression vector pPIC9K containing AOX1 promoter and the sequences of α secreting signal peptides, a recombinant expression plasmid pPIC9K/Flt-1 (1-3) was constructed and transformed to yeast host strain GS115, then His + Mut s phenotype transformant was screened out and cultured in flasks, and Flt-1 (1-3) was expressed under the induction of 1% methanol. Results SDS-PAGE showed that after being induced with 1% methanol for 4d, the expressed product existed in supernatant in the form of soluble molecule and contained 60% of total protein expressed. Western blot showed good antigenicity and specificity of expressed product. After being purified by CM-Sepharose FF and Sephacryl S-100 chromatography, the purity of the expressed product reached above 90%. Biological assay proved that the expressed product could bind to hVEGF 165 and inhibit the proliferation of HUVEC stimulated by hVEGF 165. Conclusion Human vascular endothelial growth factor receptor Flt-1 extracellular domain 1-3 loop was successfully expressed. The study lays a foundation for further application of the expressed product in the treatment of vasoformation related diseases, such as tumor and diabetic retinopathy.

关键词： Base Sequence Blotting Western Cells Cultured Cloning Molecular DNA Primers DNA Complementary Electrophoresis Polyacrylamide Gel Enzyme-Linked Immunosorbent Assay Humans Phenotype Pichia Plasmids Polymerase Chain Reaction Proto-Oncogene Proteins Receptor Protein-Tyrosine Kinases Recombinant Proteins Research Support Non-U.S. Gov't Vascular Endothelial Growth Factor Receptor-1

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Freeliving marine nematodes as a pollution indicator of the Bohai Sea

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Journal of Environmental Sciences 2002年第4期14卷 558-562页

作者： GUOYu-qing R.M.Warwick zhangzhi-nan MUFang-hong CollegeofLifeSciences OceanUniversityofQingdaoQingdao266003China CCMSPlymouthMarineLaboratory ProspectPlacePlymouthPL13DHUK

A hierarchical diversity index—taxonomic distinctness index Δ +, which was first defined by Warwick and Clark in 1998, was employed to evaluate the pollution status of the Bohai Sea with freeliving marine nematodes. The result showed that the Bohai Bay and other coastal sampling sites might be affected by oil and gas production and other anthropogenic influences. In other words,anthropogenic disturbance was affecting this component of the benthos in these locations. And most offshore sampling sites in the middle of the Bohai Sea were clear and unpolluted.

关键词： taxonomic distinctness index Δ + freeliving marine nematodes the Bohai Sea

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Bowel preparation before colonoscopy: Consequences, mechanisms, and treatment of intestinal dysbiosis

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World Journal of Gastroenterology 2025年第2期31卷 13-26页

作者： Ze-Long Zheng qing-Fan Zheng Li-Qiang Wang Yi Liu Department of Gastroenterology(Endoscopy Center) China-Japan Union Hospital of Jilin UniversityChangchun 130033Jilin ProvinceChina

The term“gut microbiota”primarily refers to the ecological community of various microorganisms in the gut,which constitutes the largest microbial community in the human *** adequate bowel preparation can improve the results of colonoscopy,it may interfere with the gut *** preparation for colonoscopy can lead to transient changes in the gut microbiota,potentially affecting an individual’s health,especially in vulnerable populations,such as patients with inflammatory bowel ***,measures such as oral pro-biotics may ameliorate these adverse *** focused on the bowel prepa-ration-induced changes in the gut microbiota and host health status,hypothesized the factors influencing these changes,and attempted to identify measures that may reduce dysbiosis,thereby providing more information for individualized bowel preparation for colonoscopy in the future.

关键词： Bowel preparation Colonoscopy Microbiota Inflammatory bowel disease Probiotics

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Effect of pyrite content on chalcopyrite flotation under different regrinding conditions

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International Journal of Minerals,Metallurgy and Materials 2025年第1期32卷 49-57页

作者： Zejun Wang qing Shi Guofan Zhang Yuxuan Zhu Binbin Li School of Minerals Processing and Bioengineering Central South UniversityChangsha 410083China Key Laboratory of Hunan Province for Clean and Efficient Utilization of Strategic Calcium-containing Mineral Resources Central South UniversityChangsha 410083China

This study aimed to investigate the effect of varying pyrite(Py)content on copper(Cu)in the presence of different regrinding conditions,which were altered using different types of grinding media:iron,ceramic balls,and their mixture,followed by flotation in the cleaner *** flotation performance of rough Cu concentrate can be improved by changing the regrinding conditions based on the Py *** electron microscopy,X-ray spectrometry,ethylenediaminetetraacetic acid disodium salt extraction,and X-ray photoelectron spectroscopy studies illustrated that when the Py content was high,the use of iron media in regrinding promoted the generation of hydrophilic Fe OOH on the surface of Py and improved the Cu *** ceramic medium with a low Py content prevented excessive Fe OOH from covering the surface of chalcopyrite(Cpy).Electrochemical studies further showed that the galvanic corrosion current of Cpy-Py increased with the addition of Py and became stronger with the participation of iron media.

关键词： chalcopyrite regrinding media pyrite galvanic corrosion flotation

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Potassium and calcium channels in different nerve cells act as therapeutic targets in neurological disorders

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Neural Regeneration Research 2025年第5期20卷 1258-1276页

作者： qing Qiu Mengting Yang Danfeng Gong Haiying Liang Tingting Chen Department of Pharmacology School of PharmacyNantong UniversityNantongJiangsu ProvinceChina Jiangsu Province Key Laboratory of Inflammation and Molecular Drug Target NantongJiangsu ProvinceChina Department of Pharmacy Longyan First Affiliated Hospital of Fujian Medical UniversityLongyanFujian ProvinceChina

The central nervous system, information integration center of the body, is mainly composed of neurons and glial cells. The neuron is one of the most basic and important structural and functional units of the central nervous system, with sensory stimulation and excitation conduction functions. Astrocytes and microglia belong to the glial cell family, which is the main source of cytokines and represents the main defense system of the central nervous system. Nerve cells undergo neurotransmission or gliotransmission, which regulates neuronal activity via the ion channels, receptors, or transporters expressed on nerve cell membranes. Ion channels, composed of large transmembrane proteins, play crucial roles in maintaining nerve cell homeostasis. These channels are also important for control of the membrane potential and in the secretion of neurotransmitters. A variety of cellular functions and life activities, including functional regulation of the central nervous system, the generation and conduction of nerve excitation, the occurrence of receptor potential, heart pulsation, smooth muscle peristalsis, skeletal muscle contraction, and hormone secretion, are closely related to ion channels associated with passive transmembrane transport. Two types of ion channels in the central nervous system, potassium channels and calcium channels, are closely related to various neurological disorders, including Alzheimer's disease, Parkinson's disease, and epilepsy. Accordingly, various drugs that can affect these ion channels have been explored deeply to provide new directions for the treatment of these neurological disorders. In this review, we focus on the functions of potassium and calcium ion channels in different nerve cells and their involvement in neurological disorders such as Parkinson's disease, Alzheimer's disease, depression, epilepsy, autism, and rare disorders. We also describe several clinical drugs that target potassium or calcium channels in nerve cells and could be u

关键词： astrocytes calcium channels central nervous system extracellular ion concentration microglia neurological disorders neurons potassium channels

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Theoretical Study on the Mechanism Properties of a Novel Trans-platinum Antitumor Drug

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Chinese Journal of Structural Chemistry 2005年第7期24卷 763-768,736页

作者： HEQin ZHOULi-Xin zhangzhi-Qiang DepartmentofChemistry JinanUniversi~.GuangzhouGuangdong510632China／／DepartmentofChemistryFuzhouUniversityFuzhouFujian350002China DepartmentofChemistry JinanUniversityGuangzhouGuangdong510632China

The mechanism properties of a novel trans-platinum (with one or two of the normal ammine ligands replaced by the planar nitrogen ligand) antitumor drug were explored by using the B3LYP method in the present paper, and the interaction for the monofunctional adduct on DNA trans-[Pt(NH3)(QUIN)GCl] (QUIN = quinoline, G = guanine) with N-containing ligands has been theoretically studied and compared with that of S-containing ligands. The results show that the N-containing ligands are more preferred in the gas phase. Environment effect has been investigated systematically using the PCM model with a series of dielectric constants ε (4.9, 9.0 and 78.4) and compared to the gas phase result with a similar trend.

关键词： .adduct LYP theoretical study ammine monofunctional trans Pt containing ligands properties of novel antitumor drug Mechanism Properties Novel Trans-platinum planar nitrogen NH QUIN

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Preparation of FeCoNi medium entropy alloy from Fe^(3+)-Co^(2+)-Ni^(2+)solution system

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International Journal of Minerals,Metallurgy and Materials 2025年第1期32卷 92-101页

作者： Zongyou Cheng qing Zhao Mengjie Tao Jijun Du Xingxi Huang Chengjun Liu Key Laboratory for Ecological Metallurgy of Multimetallic Mineral(Ministry of Education) Northeastern UniversityShenyang 110819China School of Metallurgy Northeastern UniversityShenyang 110819China Engineering Research Center of Frontier Technologies for Low-carbon Steelmaking(Ministry of Education) Shenyang 110819China

In recent years,medium entropy alloys have become a research hotspot due to their excellent physical and chemical *** controlling reasonable elemental composition and processing parameters,the medium entropy alloys can exhibit similar properties to high entropy alloys and have lower *** this paper,a FeCoNi medium entropy alloy precursor was prepared via sol-gel and coprecipitation methods,respectively,and FeCoNi medium entropy alloys were prepared by carbothermal and hydrogen *** phases and magnetic properties of FeCoNi medium entropy alloy were *** showed that FeCoNi medium entropy alloy was produced by carbothermal and hydrogen reduction at 1500℃.Some carbon was detected in the FeCoNi medium entropy alloy prepared by carbothermal *** alloy prepared by hydrogen reduction was uniform and showed a relatively high ***,the hydrogen reduction product exhibited better saturation magnetization and lower coercivity.

关键词： medium entropy alloy sol-gel co-precipitation carbothermal hydrogen reduction

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Coagulation indices and fibrinogen degradation products as predictive biomarkers for tumor-node-metastasis staging and metastasis in gastric cancer

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World Journal of Gastrointestinal Oncology 2025年第1期17卷 110-120页

作者： Yi-qing Shen Qiu-Wan Wei Yi-Ren Tian Yun-Zhi Ling Min Zhang Clinical Laboratory Civil Aviation Shanghai HospitalGubei Branch of Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of MedicineShanghai 200000China Clinical Laboratory Tongji Hospital of Tongji UniversityShanghai 200000China

BACKGROUND Gastric cancer(GC)is a prevalent malignancy with a substantial health burden and high mortality rate,despite advances in prevention,early detection,and *** with the global average,Asia,notably China,reports disproportionately high GC *** disease often progresses asymptoma-tically in the early stages,leading to delayed diagnosis and compromised ***,it is crucial to identify early diagnostic biomarkers and enhance treatment strategies to improve patient outcomes and reduce *** Retrospectively analyzed the clinical data of 148 patients with GC treated at the Civil Aviation Shanghai Hospital between December 2022 and December *** associations of coagulation indices-partial thromboplastin time(APTT),prothrombin time(PT),thrombin time(TT),fibrinogen,fibrinogen degradation products(FDP),fasting blood glucose,and D-dimer(D-D)with TNM stage and distant metastasis were *** Prolongation of APTT,PT,and TT was significantly correlated with the GC TNM ***,abnormal coagulation system activation was closely related to disease *** FDP and D-D were significantly associated with distant metastasis in GC(P<0.05),suggesting that increased fibrinolytic activity contributes to increased metastatic *** Our Results reveal coagulation indices,FDPs as GC biomarkers,reflecting abnormal coagulation/fibrinolysis,aiding disease progression,metastasis prediction,and helping clinicians assess thrombotic risk for early intervention and personalized treatment plans.

关键词： Coagulation indexes Fibrinogen degradation products Gastric cancer Tumor-node-metastasis staging Distant metastasis

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The 2D Alternative Binary L—J System：Solid—Liquid Phase Diagram

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Communications in Theoretical Physics 2002年第5期37卷 601-606页

作者： zhangzhi CHENLi－Rong DepartmentofPhysics AnhuiNormalUniversityWuhu241000AnhuiProvinceChina

The Lennard-Jones potential is introduced into the Collins model and is generalized to the two-dimensionalalternative binary system. The Gibbs free energy of the binary system is calculated. According to the thermodynamicconditions of solid-liquid equilibrium, the "cigar-type" phase diagram and the phase diagram with a minimum areobtained. The results are quite analogous to the behavior of three-dimensional substances.

关键词：热力学固态-液态相图 2DBernal模型 L-J势

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