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Chemical Research in Chinese Universities 2011年第3期27卷 450-454页

作者： ZHAO Hai-feng WU Lei LU Ming HU Yu-lin HAN Xiao LI Zheng-qiang Key Laboratory for molecular Enzymology ＆ Engineering Ministry of Education Jilin University Changchun 130012 P. R. China Medical department of Liver Gall Pancreas the First Hospital of Jilin University Changchun 130021 P. R. China

Lipid-bound [VHb（406）] and lipid-free [VHb（402）] wide type Vitreoscilla hemoglobin were separated from E. coli BL21（DE3）. The RZ is 3.30 for [VHb（406）] and 3.15 for [VHb（402）], respectively. VHb（406） shows a characteristic absorption band at 626 nm, while VHb（402） shows one at 644 nm, and it has more α-helix than VHb（402）. Data of Raman spectra experiment shows under the excitation wavelength 488 nm, υ4 vibration of both VHb（402） and VHb（406） had a pure and strong signal at 1375 cm-1, which proves that iron porphyrin of both the samples is at their trivalence oxidation state. And no matter lipid binds VHb or not, the vinyl of porphyrin is at cis state. The catalase activity of Vitreoscilla hemoglobin was explored with L-dopa and H2O2 as substrates. The results indicate that VHb（406） has more catalase activity than VHb（402）, VHb is a cell’s oxygen modulator and its catalase ability is modulated by the membrane.

关键词： Vitreoscilla hemoglobin Structure Function Membrane binding

In vitro Study of Nucleostemin Gene as a Potential Therapeutic Target for Human Lung Carcinoma

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Biomedical and Environmental Sciences 2012年第1期25卷 91-97页

作者： GAO Hong Xia GAO Xiu Feng WANG Guo Qing WANG En Shu HUANG Wei HUANG Ping department of Biochemistry and molecular Biology West China School of Preclinical and Forensic Medicine Sichuan University Chengdu 610041 Sichuan China Institute of Laboratory Medicine Beihua University Jilin 132013 Jilin China College of Communication Engineering Jilin University Changchun 130012 Jilin China

Objective Nucleostemin （NS） is a GTP-conjugated protein located in the nucleoli of stem cells and some cancer cells, and maintains cell self-renewal. We aimed to evaluate NS as a potential target for lung carcinoma gene therapy by investigating NS gene expression and its effect on A549 cell proliferation. Methods NS mRNA and protein expression in A549, HepG2, SMMC-7721, HeLa, and U251 cells was analyzed by RT-PCR and western blotting following transfection of NS siRNAs and negative control siRNA （NC）. The effect on cell proliferation was also analyzed by MTF assays. Results NS mRNA and protein were both expressed in A549 cells and four other tumor cell lines; the relative expression levels were similar in all five cell lines. The three pairs of NS siRNA, either transfected alone or cotransfected into A549 cells, could effectively inhibit the expression of NS mRNA and protein. Moreover, the interference ratio showed an obvious concentration-dependent relationship. NS siRNA treatment resulted in significant inhibition of A549 cell proliferation by 35.7%. Conclusion NS gene was not only highly expressed but also played an important role in A549 cell proliferation. Thus, targeting of NS may be a promising novel strategy for the treatment of lung carcinoma.

关键词： Nucleostemin RNA interference A549 cells Tumor gene therapy

Müller cells are activated in response to retinal outer nuclear layer degeneration in rats subjected to simulated weightlessness conditions

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Neural Regeneration Research 2025年第7期20卷 2116-2128页

作者： Yuxue Mu Ning Zhang Dongyu Wei Guoqing Yang Lilingxuan Yao Xinyue Xu Yang Li Junhui Xue Zuoming Zhang Tao Chen Aerospace Clinical Medical Center School of Aerospace MedicineAir Force Medical UniversityXi’anShaanxi ProvinceChina department of Aviation Medicine Xijing HospitalAir Force Medical UniversityXi’anShaanxi ProvinceChina department of Emergency Medicine Wuhan No.1 HospitalWuhanHubei ProvinceChina Third Regiment School of Basic MedicineAir Force Medical UniversityXi’anShaanxi ProvinceChina Fourth Regiment School of Basic MedicineAir Force Medical UniversityXi’anShaanxi ProvinceChina

A microgravity environment has been shown to cause ocular damage and affect visual acuity,but the underlying mechanisms remain ***,we established an animal model of weightlessness via tail suspension to examine the pathological changes and molecular mechanisms of retinal damage under *** 4 weeks of tail suspension,there were no notable alterations in retinal function and morphology,while after 8 weeks of tail suspension,significant reductions in retinal function were observed,and the outer nuclear layer was thinner,with abundant apoptotic *** investigate the mechanism underlying the degenerative changes that occurred in the outer nuclear layer of the retina,proteomics was used to analyze differentially expressed proteins in rat retinas after 8 weeks of tail *** results showed that the expression levels of fibroblast growth factor 2(also known as basic fibroblast growth factor)and glial fibrillary acidic protein,which are closely related to Müller cell activation,were significantly *** addition,Müller cell regeneration and Müller cell gliosis were observed after 4 and 8 weeks,respectively,of simulated *** findings indicate that Müller cells play an important regulatory role in retinal outer nuclear layer degeneration during weightlessness.

关键词： glial fibrous acidic protein gliosis Müller cells nerve growth factor neural differentiation neurodegeneration proteomic retinal degeneration retinal outer nuclear layer simulated weightlessness

Telmisartan induces melanoma cell apoptosis and synergizes with vemurafenib in vitro by altering cell bioenergetics

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Cancer Biology & Medicine 2019年第2期16卷 247-263页

作者： Jelena Grahovac Tatjana Srdic-Rajic Juan Francisco Santibanez Marijana Pavlovic Milena Cavic Sinisa Radulovic1 Laboratory for Experimental Pharmacology Institute for Oncology and Radiology of Serbia Belgrade 11000 Serbia department of molecular Oncology Institute for Medical Research University of Belgrade Belgrade 11000 Serbia Integrative Center for Biology and Applied Chemistry (CIBQA) Bernardo O'Higgins University Santiago 8370854 Chile

Objective: Despite recent advancements in targeted therapy and immunotherapies, prognosis for metastatic melanoma patients remains extremely poor. Development of resistance to previously effective treatments presents a serious challenge and new approaches for melanoma treatment are urgently needed. The objective of this study was to examine the effects of telmisartan, an AGTR1 inhibitor and a partial agonist of PPARγ, on melanoma cells as a potential agent for repurposing in melanoma ***: Expression of AGTR1 and PPARγ m RNA in melanoma patient tumor samples was examined in publicly available datasets and confirmed in melanoma cell lines by qRT-PCR. A panel of melanoma cell lines was tested in viability, apoptosis and metabolic assays in presence of telmisartan by flow cytometry and immunocytochemistry. A cytotoxic effect of combinations of telmisartan and targeted therapy vemurafenib was examined using the Chou-Talalay combination index ***: Both AGTR1 and PPARγ mRNA were expressed in melanoma patient tumor samples and decreased compared to the expression in the healthy skin. In vitro, we found that telmisartan decreased melanoma cell viability by inducing cell *** glucose uptake, but not utilization, in the presence of telmisartan caused the fission of mitochondria and release of reactive oxygen species. Telmisartan altered the cell bioenergetics, thereby synergizing with vemurafenib in vitro, and even sensitized vemurafenib-resistant cells to the ***: Given that the effective doses of telmisartan examined in our study can be administered to patients and that telmisartan is a widely used and safe antihypertensive drug, our findings provide the scientific rationale for testing its efficacy in treatment of melanoma progression.

关键词： Melanoma telmisartan apoptosis mitochondria reactive oxygen species targeted therapy

Sirtuins in B lymphocytes metabolism and function

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World Journal of Experimental Medicine 2019年第1期9卷 1-13页

作者： Bruno Ghirotto Fernanda Fernandes Terra Niels Olsen Saraiva Camara Paulo José Basso department of Immunology Institute of Biomedical Sciences University of Sao Paulo Division of Nephrology School of Medicine Federal University of Sao Paulo Laboratory of Renal physiology (LIM 16) School of MedicineUniversity of S?o Paulo

Sirtuins(SIRTs) are NAD+-dependent histone deacetylases and play a role in virtually all cell biological processes. As SIRTs functions vary according to their subtypes, they can either activate or inhibit signaling pathways upon different conditions or tissues. Recent studies have focused on metabolic effects performed by SIRTs in several cell types since specific metabolic pathways(e.g., aerobic glycolysis, oxidative phosphorylation, β-oxidation, glutaminolysis) are used to determine the cell fate. However, few efforts have been made to understand the role of SIRTs on B lymphocytes metabolism and function. These cells are associated with humoral immune responses by secreting larger amounts of antibodies after differentiating into antibody-secreting cells. Besides, both the SIRTs and B lymphocytes are potential targets to treat several immune-mediated disorders, including cancer. Here, we provide an outlook of recent studies regarding the role of SIRTs in general cellular metabolism and B lymphocytes functions, pointing out the future perspectives of this field.

关键词： B cells Metabolic sensors Histone deacetylases Cancer

molecular Basis for Sesterterpene Diversity Produced by Plant Terpene Synthases

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Plant Communications 2020年第5期1卷 41-52页

作者： Qingwen Chen Jianxu Li Zhixi Liu Takaaki Mitsuhashi Yuting Zhang Haili Liu Yihua Ma Juan He Tetsuro Shinada Tsutomu Sato Yong Wang Hongwei Liu Ikuro Abe Peng Zhang Guodong Wang State Key Laboratory of Plant Genomics and National Center for Plant Gene Research Institute of Genetics and Developmental BiologyThe Innovative Academy of Seed DesignChinese Academy of SciencesBeijing 100101China National Key Laboratory of Plant molecular Genetics Center for Excellence in Molecular Plant SciencesShanghai Institute of Plant Physiology and EcologyChinese Academy of SciencesShanghai 200032China Graduate School of Pharmaceutical Sciences University of Tokyo7-3-1 HongoBunkyo-kuTokyo 113-0033Japan State Key Laboratory of Mycology Institute of MicrobiologyChinese Academy of SciencesBeijing 100101China Key Laboratory of Synthetic Biology Institute of Plant Physiology and EcologyShanghai Institutes for Biological SciencesChinese Academy of SciencesShanghai 200032China University of Chinese Academy of Sciences Beijing 100039China Graduate School of Science Osaka City University3-3-138 SugimotoSumiyoshiOsaka 558-8585Japan department of Applied Biological Chemistry Faculty of AgricultureGraduate School of Science and TechnologyNiigata University8050 Ikarashi-2Niigata 950-2181Japan

Class I terpene synthase(TPS)generates bioactive terpenoids with diverse *** synthase(sester-TPS,C25),a branch of class I TPSs,was recently identified in ***,the catalytic mechanisms of sester-TPSs are not fully ***,we first identified three nonclustered functional sester-TPSs(AtTPS06,AtTPS22,and AtTPS29)in Arabidopsis ***06 utilizes a type-B cyclization mechanism,whereas most other sester-TPSs produce various sesterterpene backbones via a type-A cyclization *** then determined the crystal structure of the AtTPS18–FSPP complex to explore the cyclization mechanism of plant *** used structural comparisons and site-directed mutagenesis to further elucidate the mechanism:(1)mainly due to the outward shift of helix G,plant sester-TPSs have a larger catalytic pocket than do mono-,sesqui-,and di-TPSs to accommodate GFPP;(2)type-A sester-TPSs have more aromatic residues(five or six)in their catalytic pocket than classic TPSs(two or three),which also determines whether the type-A or type-B cyclization mechanism is active;and(3)the other residues responsible for product fidelity are determined by interconversion of AtTPS18 and its close ***,this study improves our understanding of the catalytic mechanism of plant sester-TPS,which ultimately enables the rational engineering of sesterterpenoids for future applications.

关键词： terpenoid terpene synthase sesterterpene cyclization mechanism crystal structure

Characterization of SARS-CoV-2-specific antibodies in COVID-19 patients reveals highly potent neutralizing IgA

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Signal Transduction and Targeted Therapy 2021年第2期6卷 254-256页

作者： Weihong Zeng Huan Ma Chengchao Ding Yunru Yang Yong Sun Xiaoxue Huang Weihuang He Yan Xiang Yong Gao Tengchuan Jin Division of Life Sciences and Medicine Department of Obstetrics and GynecologyThe First Afliated Hospital of USTCUniversity of Science and Technology of ChinaHefeiAnhuiChina Hefei National Laboratory for Physical Sciences at Microscale CAS Key Laboratory of Innate Immunity and Chronic DiseaseDivision of Life Sciences and MedicineSchool of Basic Medical SciencesUniversity of Science and Technology of ChinaHefeiAnhuiChina Division of Life Sciences and Medicine Department of Pulmonary and Critical Care MedicineThe First Affliated Hospital of USTCUniversity of Science and Technology of ChinaHefeiAnhuiChina Division of Life Sciences and Medicine Department of Infectious DiseasesThe First Affliated Hospital of USTCUniversity of Science and Technology of ChinaHefeiAnhuiChina Anhui Center for Disease Control and Prevention(Anhui CDC) HefeiAnhuiChina Kangrun Biotech LTD GuangzhouGuangdongChina department of Microbiology Immunology and Molecular GeneticsUniversity of Texas Health Science Center at San AntonioSan AntonioTXUSA CAS Center for Excellence in molecular cell Science Chinese Academy of ScienceShanghaiChina

Dear Editor,The coronavirus disease 2019(COVID-19),caused by the Severe Acute Respiratory Syndrome Coronavirus 2(SARS-CoV-2),is an ongoing threat to public *** to the presence of neutralizing antibodies in convalescent patients,convalescent plasma transfusion has been tested as a promising therapy for severe COVID-19 patients.12 We and others previously found a surprisingly early serum lgA antibody response in the COVID-19 ***,the relative contribution of the lgA antibody towards virus neutralization with respect to other antibody isotypes is unknown.

关键词： patients antibodies transfusion

Tumor cell-specific gene transfer with retroviral vectors displaying single-chain antibody

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Chinese Medical Journal 2002年第7期115卷 1064-1069,153页

作者：汤宇澄李煜钱关祥 department of Biochemistry and molecular Biology the Researeh Center for Human Gene TherapyShanghai Second Medical UniversityShanghai 200025China

OBJECTIVE: To specifically deliver the therapeutic gene to cancer cells and construct target retroviral vectors by inserting the single-chain variable antibody fragment into the retroviral envelope. METHODS: Single-chain antibody expression vector pET -20bScfv was constructed. Binding activity of the genetically engineered single-chain variable antibody fragment was verified by enzyme-linked immunosorbent assay (ELISA) and Western blot. At the same time, by means of polymerase chain reaction (PCR)-directed mutagenesis, the appropriate cloning site EcoT22/Sal was generated at the N-terminus of receptor-binding SU domain in the MoMLV env polypeptide. Then the single- chain antibody gene, encoding a functional antibody, was inserted into the cloning site. The Scfv-env fusion gene fragment was subcloned into CMV expression vector. The Lac-Z retrovirus that co-displayed the Scfv-env chimeric protein and wild-type env protein was produced by transfection of Psi 2 cells with retroviral plasmid and the fusion gene expressing *** confirm the specificity of the recombinant retrovirus, infection assays and competitive inhibition assays were performed. RESULTS: The results of ELISA and Western blot showed that the genetically engineered single-chain variable antibody fragment could bind to the SHG44 cells surface membrane antigen. Virus-binding assay, viral infection and competitive inhibition assays confirmed that the harvested virus efficiently bound to and infected SHG44 cancer cells expressing the relative membrane antigen specially via the recognition of the target antigen. CONCLUSION: These results imply that insertion of Scfv into the retroviral envelope can specifically deliver the interested gene into specific antigen-producing cancer cells.

关键词： Gene Therapy Gene Transfer Techniques Genetic Vectors Hela cells Humans Immunoglobulin Fragments Neoplasms Research Support, Non-U.S. Gov't Retroviridae

Functional maturation of outer hair cells in the rat cochlea

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Functional maturation of outer hair cells in the rat cochlea