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Expression, Purification and Sublocalization of SARS-CoV Nucleocapsid Protein in Insect Cells

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Acta Biochimica et Biophysica Sinica 2004年第11期36卷 754-758页

作者： Ai-Xia REN, You-Hua XIE, Yu-Ying KONG, Guan-Zhen YANG, yao-zhou zhang,Yuan WANG, and xiang-fu wuinstitute of biochemistry and Biotechnology, college of life Science, zhejiang university, hangzhou 310029, china State Key Laboratory of Molecular Biology, Shanghai institute of biochemistry and cell Biology, Shanghai institutes for Biological sciences,Chinese Academy of sciences, Graduate School of the Chinese Academy of sciences, Shanghai 200031, china institute of biochemistry, zhejiang university of sciences, hangzhou 310018, china institute of biochemistry and Biotechnology College of Life Science Zhejiang University Hangzhou 310029 China.

The causative agent of severe acute respiratory syndrome (SARS) is a previously unidentified coronavirus, SARS-CoV. The nucleocapsid (N) protein of SARS-CoV is a major viral protein recognized by acute and early convalescent sera from SARS patients. To facilitate the studies on the function and structure of the N protein, this report describe the expression and purification of recombinant SARS-CoV N protein using the baculovirus

关键词： severe acute respiratory syndrome (SARS) SARS coronavirus nucleocapsid protein baculovirus insect cells

Embryo Transfer Strategies for Women with Recurrent Implantation Failure During the Frozen-thawed Embryo Transfer Cycles:Sequential Embryo Transfer or Double-blastocyst Transfer?

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Current Medical Science 2024年第1期44卷 212-222页

作者： Qiao-hang ZHAO Yu-wei SONG Jian CHEN xiang zhou Ji-lai XIE Qiu-ping yao Qi-yin DONG Chun FENG Li-ming zhou Wei-ping fu Min JIN The Second Affiliated Hospital of zhejiang university School of Medicine Hangzhou 310052China Jiaxing Maternity and Child Health Care Hospital Jiaxing 314051China Huzhou Maternity and Child Care Hospital Huzhou 313002China Ningbo Women and Children's Hospital Ningbo 315000China

Objective Both sequential embryo transfer(SeET)and double-blastocyst transfer(DBT)can serve as embryo transfer strategies for women with recurrent implantation failure(RIF).This study aims to compare the effects of SeET and DBT on pregnancy *** Totally,261 frozen-thawed embryo transfer cycles of 243 RIF women were included in this multicenter retrospective *** to different embryo quality and transfer strategies,they were divided into four groups:group A,good-quality SeET(GQ-SeET,n=38 cycles);group B,poor-quality or mixed-quality SeET(PQ/MQ-SeET,n=31 cycles);group C,good-quality DBT(GQ-DBT,n=121 cycles);and group D,poor-quality or mixed-quality DBT(PQ/MQ-DBT,n=71 cycles).The main outcome,clinical pregnancy rate,was compared,and the generalized estimating equation(GEE)model was used to correct potential confounders that might impact pregnancy *** GQ-DBT achieved a significantly higher clinical pregnancy rate(aOR 2.588,95%CI 1.267–5.284,P=0.009)and live birth rate(aOR 3.082,95%CI 1.482–6.412,P=0.003)than PQ/***,the clinical pregnancy rate was significantly higher in GQ-SeET than in PQ/MQ-SeET(aOR 4.047,95%CI 1.218–13.450,P=0.023).The pregnancy outcomes of GQ-SeET were not significantly different from those of GQ-DBT,and the same results were found between PQ/MQ-SeET and PQ/*** SeET relative to DBT did not seem to improve pregnancy outcomes for RIF patients if the embryo quality was comparable between the two *** clinical pregnancy outcomes could be obtained by transferring good-quality embryos,no matter whether in SeET or *** quality plays a more important role in pregnancy outcomes for RIF patients.

关键词： recurrent implantation failure sequential embryo transfer frozen-thawed embryo transfer embryo transfer strategies

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家蚕核型多角体病毒遍在蛋白及结合肽的研究

生物化学与生物物理进展 2004年第11期31卷 1038-1044页

作者：张文波林蓉张耀洲吴祥甫浙江大学生命科学学院浙江理工大学生物化学研究所杭州310018 浙江大学动物科学学院浙江理工大学生物化学研究所中国科学院生物化学与细胞生物学研究所

从家蚕核型多角体病毒镇江株 (BombyxmorinuclearpolyhedrosisvirusZhenjiangstrain ,BmNPV ZJ)基因组DNA中克隆遍在蛋白 (BmVUB)基因 .序列分析结果显示 ,BmVUB基因长 2 34bp ,编码 77个氨基酸 .BmVUB氨基酸序列内有一致HTH序列 ,遍在... 详细信息

从家蚕核型多角体病毒镇江株 (BombyxmorinuclearpolyhedrosisvirusZhenjiangstrain ,BmNPV ZJ)基因组DNA中克隆遍在蛋白 (BmVUB)基因 .序列分析结果显示 ,BmVUB基因长 2 34bp ,编码 77个氨基酸 .BmVUB氨基酸序列内有一致HTH序列 ,遍在蛋白保守序列LRLRGG ,参与遍在蛋白蛋白酶复合体形成的 4个保守性功能位点 (Lys 2 9、Cys 4 8、Cys 6 3、Gly 76 )及保守的Gly Gly X (X是疏水氨基酸残基 )蛋白酶切信号序列 ,在遍在蛋白保守的Gly Gly后多出由一个氨基酸组成的延伸肽 .原核诱导表达表明BmVUB主要以可溶性形式存在 ,表达量占总菌蛋白的 5 0 %以上 .纯化的遍在蛋白浓度为 1 0 3～ 2 4 6 g/L ,制备抗体 ,效价在 3 2× 10 -5以上 .用噬菌体表面展示技术筛选遍在蛋白结合肽 ,所筛选的结合肽可与遍在蛋白特异性结合 ,其中结合肽VAPHHAYAPMRT对细胞增殖有明显的浓度调控作用 ,即低浓度促进细胞生长 ,高浓度强烈抑制细胞的生长 .

关键词：结合肽遍在蛋白白及性功能原核细胞增殖细胞生长诱导表达氨基酸序列特异性结合

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Expression of Human Lactoferrin in B. mori-cultured Cells and Its Bacteriostatic Activity

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Expression of Human Lactoferrin in B. mori-cultured Cells an...

Expression and Activity Determination of Recombinant Osteoprotegerin from Silkworm Larvae

浙江省生物化学与分子生物学学术交流会

作者： liu tao , zhang yao-zhou , wu xiang-fu institute of biochemistry, college of life sciences, zhejiang university, hangzhou 310029, china. institute of biochemistry, zhejiang Sci-Tec university, hangzhou 310018, china. institute of biochemistry and cell Biology, Chinese Academy of sciences, Shanghai 200031, china

Human lactoferrin ( hLf ) is a multifunctional iron-binding glycoprotin. In this study, we amplified hLf cDNA by RT-PCR from normal human mammary gland. The N-terminal sequence of the hLf was deduced as GRRRRSV which was identical to the known hLf. We constructed a recombinant virus vBm-hLf harboring hLf gene and exploited the BmN cells as host to produce recombinant human lactoferrin (rhLf ). It was found that a -78 kDa recombinant protein was expressed. Around 13.5 mg/L of rhLf was purified from BmN cells culture medium infected by vBm-hLf and the rhLf was proved to be biologically active. This method established in our study will pave the way for efficient production of rhLf for further utilization of this protein in the future.

关键词： human lactoferrin RT-PCR bombyx mori cultured cells bacteriostatic activity.

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Expression and Activity Determination of Recombinant Osteopr...

Two-dimensional materials for future information technology: status and prospects

首届长三角科技论坛——长三角生物医药发展论坛

作者： Hal-Long XIAO yao-zhou zhang~1 xiang-fu wu~2 college of life sciences,zhejiang university,hangzhou 310029,china ~1institute of biochemistry,college of life sciences,zhejiang university of sciences,hangzhou 310018,china ~2institute of biochemistry and Cell Biology,Shanghai institute for Biological sciences,C A S,Shanghai 200025,china

Osteoprotegerin(OPG)regulates the formation of osteoclasts and is involved in the regulation ofbone resorption and *** order to investigate the feasibility of using silkworm(Bombyx mori)larvae toproduce recombinant osteoprotegerin,we constructed recombinant Bombyx mori nuclear polyhedrosis viruses(BmNPV)canting osteoprotegerin cDNA,and expressed rhOPG in the larvae of silkworm by inoculation ofrecombinant *** expression product was run on the SDS-PAGE and its immunoreactivitiy was determinedby Western *** was found that 47kD recombinant protein was expressed in *** silkwormhemolymph prepared from larvae infected with recombinant virus was injected to mice abdominal cavity,Compared with the control,this injection was effective to decrease serum calcium concentration,and increasethe percent trabecular bone area in the distal femurs of *** results showed that rhOPG expressed insilkworm larvae had the proper ***:osteoprotegerin Baculovirus expression bioactivity

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Science china(Information sciences) 2024年第6期67卷 1-147页

作者： Hao QIU Zhihao YU Tiange ZHAO Qi zhang Mingsheng XU Peifeng LI taotao LI Wenzhong BAO Yang CHAI Shula CHEN Yiqi CHEN Hui-Ming CHENG Daoxin DAI Zengfeng DI Zhuo DONG Xidong DUAN Yuhan FENG Yu fu Jingshu GUO Pengwen GUO Yue HAO Jun HE Xiao HE Jingyi HU Weida HU Zehua HU Xinyue HUANG Ziyang HUANG Ali IMRAN Ziqiang KONG Jia LI Qian LI Weisheng LI Lei LIAO Bilu liu Can liu Chunsen liu Guanyu liu Kaihui liu Liwei liu Sheng liu Yuan liu Donglin LU Likuan MA Feng MIAO Zhenhua NI Jing NING Anlian PAN Tian-Ling REN Haowen SHU Litao SUN Yue SUN Quanyang tao Zi-Ao TIAN Dong WANG Hao WANG Haomin WANG Jialong WANG Junyong WANG Wenhui WANG Xingjun WANG Yeliang WANG Yuwei WANG Zhenyu WANG yao WE... School of Electronic Science and Engineering Nanjing University School of Integrated Circuits Interdisciplinary Research Center for Future Intelligent Chips (Chip-X)Nanjing University School of Integrated Circuit Science and Engineering Nanjing University of Posts and Telecommunications State Key Laboratory of Infrared Physics Shanghai Institute of Technical Physics Chinese Academy of Sciences School of Physics and Key Lab of Quantum Materials and Devices of the Ministry of Education School of Electronic Science & EngineeringSoutheast University college of Integrated Circuits State Key Laboratory of Silicon and Advanced Semiconductor MaterialsZhejiang University Suzhou Laboratory State Key Laboratory of Integrated Chips and Systems School of MicroelectronicsFudan University Shaoxin Laboratory Department of Applied Physics The Hong Kong Polytechnic University Key Laboratory for Micro-Nano Physics and Technology of Hunan Province College of Materials Science and EngineeringHunan University college of Integrated Circuits Zhejiang University Shenzhen Geim Graphene Center Shenzhen Key Laboratory of Layered Materials for Value-Added ApplicationsTsinghua-Berkeley Shenzhen Institute & Institute of Materials ResearchTsinghua Shenzhen International Graduate SchoolTsinghua University State Key Laboratory of Extreme Photonics and Instrumentation College of Optical Science and EngineeringInternational Research Center for Advanced PhotonicsZijingang CampusZhejiang University National Key Laboratory of Materials for Integrated Circuits Shanghai Institute of Microsystem and Information TechnologyChinese Academy of Sciences CAS Key Laboratory of Nanophotonic Materials and Devices & Key Laboratory of Nanodevices and Applications i-LabSuzhou Institute of Nano-Tech and Nano-Bionics (SINANO)Chinese Academy of Sciences college of Chemistry and Chemical Engineering Hunan University Key Laboratory of Quantum State Construction and Manipulation (Ministry of Education) Department of PhysicsRenmin University of China School

Over the past 70 years, the semiconductor industry has undergone transformative changes,largely driven by the miniaturization of devices and the integration of innovative structures and materials. Two-dimensional(2D) materials like transition metal dichalcogenides(TMDs) and graphene are pivotal in overcoming the limitations of silicon-based technologies, offering innovative approaches in transistor design and functionality, enabling atomic-thin channel transistors and monolithic 3D integration. We review the important progress in the application of 2D materials in future information technology, focusing in particular on microelectronics and optoelectronics. We comprehensively summarize the key advancements across material production, characterization metrology, electronic devices, optoelectronic devices, and heterogeneous integration on silicon. A strategic roadmap and key challenges for the transition of 2D materials from basic research to industrial development are outlined. To facilitate such a transition, key technologies and tools dedicated to 2D materials must be developed to meet industrial standards, and the employment of AI in material growth, characterizations, and circuit design will be essential. It is time for academia to actively engage with industry to drive the next 10 years of 2D material research.

关键词： two-dimensional (2D) materials information technology production technology standardization of characterization microelectronicdevices optoelectronic devices multi-functional integration

人干扰素α2b-胸腺肽α1融合基因在家蚕细胞中的表达和活性研究

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生物化学与生物物理进展 2003年第5期30卷 803-807页

作者：郭冬生朱成钢张耀洲吴祥甫浙江大学生命科学院生物化学研究所杭州310029 中国科学院生物化学和细胞生物学研究所上海200031

根据细胞因子协同作用的特点,采用重组DNA技术构建了人干扰素(IFN)α2b 胸腺肽(THY)α1融合基因,克隆到pBacPAK8上,获得重组转移载体pBacPAK IFN THY.与线形化Bm BacPAK6病毒基因组DNA共转染家蚕细胞,经过体内重组,筛选到重组病毒Bm Bac... 详细信息

根据细胞因子协同作用的特点,采用重组DNA技术构建了人干扰素(IFN)α2b 胸腺肽(THY)α1融合基因,克隆到pBacPAK8上,获得重组转移载体pBacPAK IFN THY.与线形化Bm BacPAK6病毒基因组DNA共转染家蚕细胞,经过体内重组,筛选到重组病毒Bm BacPAK IFN THY.将Bm BacPAK IFN THY感染家蚕细胞进行表达.DNA印迹证明IFN THY已插入Bm BacPAK6中(4kb左右的杂交带);SDS 聚丙烯酰胺凝胶电泳、蛋白质印迹证明IFN THY在家蚕细胞中得到了表达(分子质量为 2 3ku左右),且具有IFN蛋白的免疫原性;微量细胞病变抑制法和玫瑰花结法显示 96h的表达产物IFN活性为 3 72× 10 4U/ml,12 0h表达产物IFN活性为 3 10× 10 5U/ml,4 8～ 72h表达产物IFN活性较低;4 8～ 12 0h表达产物的玫瑰花结形成率均在 10 %以上.结果表明融合基因在家蚕细胞中得到了高效表达,表达的融合蛋白具有IFN α2b和THY

关键词：人干扰素α2b-胸腺肽α1融合基因家蚕细胞活性家蚕核型多角体病毒基因表达

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Characterization of acute-on-chronic liver diseases: A multicenter prospective cohort study

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World Journal of Hepatology 2024年第5期16卷 809-821页

作者： Yuan-yao zhang Sen Luo Hai Li Shu-Ning Sun Xian-Bo Wang Xin Zheng Yan Huang Bei-Ling Li Yan-Hang Gao Zhi-Ping Qian Feng liu Xiao-Bo Lu Jun-Ping liu Hao-Tang Ren Yu-Bao Zheng Hua-Dong Yan Guo-Hong Deng Liang Qiao Yan zhang Wen-Yi Gu Xiao-Mei xiang Yi zhou Yi-Xin Hou Qun zhang Yan Xiong Cong-Cong Zou Jun Chen Ze-Bing Huang Xiu-Hua Jiang Ting-Ting Qi Yuan-Yuan Chen Na Gao Chun-Yan liu Wei Yuan Xue Mei Jing Li tao Li Rong-Jiong Zheng Xin-Yi zhou Jun Zhao Zhong-Ji Meng Department of Infectious Disease Institute of Biomedical ResearchRegulatory Mechanism and Targeted Therapy for Liver Cancer Shiyan Key LaboratoryHubei provincial Clinical Research Center for Precise Diagnosis and Treatment of Liver CancerTaihe HospitalHubei University of MedicineShiyan 442000Hubei ProvinceChina Department of Gastroenterology Renji HospitalSchool of MedicineShanghai Jiao Tong UniversityShanghai 200120China Department of Infectious Diseases Southwest HospitalThe Third Military Medical University(Army Medical University)Chongqing 400000China Center of Integrative Medicine Beijing Ditan HospitalCapital Medical UniversityBeijing 100020China Department of Infectious Diseases Institute of Infection and ImmunologyUnion HospitalTongji Medical CollegeHuazhong University of Science and TechnologyWuhan 430020Hubei ProvinceChina Department of Infectious Diseases Hunan Key Laboratory of Viral HepatitisXiangya HospitalCentral South UniversityChangsha 110051Hunan ProvinceChina Hepatology Unit Department of Infectious DiseasesNanfang HospitalSouthern Medical UniversityGuangzhou 510515Guangdong ProvinceChina Department of Hepatology The First Hospital of Jilin UniversityChangchun 130021Jilin ProvinceChina Department of Liver Intensive Care Unit Shanghai Public Health Clinical CentreFudan UniversityShanghai 200093China Tianjin institute of Hepatology Nankai University Second People's HospitalTianjin 300192China Infectious Disease Center The First Affiliated Hospital of Xinjiang Medical UniversityUrumqi 830054Xinjiang Uygur Autonomous RegionChina Department of Infectious Diseases and Hepatology The Second Hospital of Shandong UniversityJinan 250000Shandong ProvinceChina State Key Laboratory for Diagnosis and Treatment of Infectious Diseases Collaborative Innovation Center for Diagnosis and Treatment of Infectious DiseaseThe First Affiliated HospitalZhejiang University School of MedicineHangzhou 310000Zhejiang ProvinceChina Department of Infectious Diseases The Thir

BACKGROUND Acute-on-chronic liver disease(AoCLD)accounts for the majority of patients hospitalized in the Department of Hepatology or Infectious *** To explore the characterization of AoCLD to provide theoretical guidance for the accurate diagnosis and prognosis of *** Patients with AoCLD from the Chinese Acute-on-Chronic Liver Failure(ACLF)study cohort were included in this *** clinical characteristics and outcomes,and the 90-d survival rate associated with each clinical type of AoCLD were analyzed,using the Kaplan-Meier method and the log-rank *** A total of 3375 patients with AoCLD were enrolled,including 1679(49.7%)patients with liver cirrhosis acute decompensation(LC-AD),850(25.2%)patients with ACLF,577(17.1%)patients with chronic hepatitis acute exacer-bation(CHAE),and 269(8.0%)patients with liver cirrhosis active phase(LC-A).The most common cause of chronic liver disease(CLD)was HBV infection(71.4%).The most common precipitants of AoCLD was bacterial infection(22.8%).The 90-d mortality rates of each clinical subtype of AoCLD were 43.4%(232/535)for type-C ACLF,36.0%(36/100)for type-B ACLF,27.0%(58/215)for type-A ACLF,9.0%(151/1679)for LC-AD,3.0%(8/269)for LC-A,and 1.2%(7/577)for *** HBV infection is the main cause of CLD,and bacterial infection is the main precipitant of *** most common clinical type of AoCLD is *** diagnosis and timely intervention are needed to reduce the mortality of patients with LC-AD or ACLF.

关键词： Acute-on-chronic liver disease Acute-on-chronic liver failure Liver cirrhosis Clinical features Prognosis

Cu-catalyzed cross-couplings under ligandless conditions

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Chinese Chemical Letters 2007年第10期18卷 1199-1202页

作者： Chuan zhou tao Juan Li Xin Cui yao fu Qing xiang Guo Department of Chemistry University of Science and Technology of China Hefei 230026 China

The possibility to carry out Cu-catalyzed cross-couplings under ligandless conditions was studied. It was found that by using K3PO4 as the base, the cross-coupling could proceed successfully between aryl iodides and s... 详细信息

关键词： Cu Catalysis Cross-coupling Ligand free

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