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Expression and purification of lipoprotein-associated phospholipase A2,a key enzyme involved in atherosclerosis

Acta Pharmacologica Sinica 2006年第6期27卷 679-684页

作者： Fu-jun ZHANG~(2,4)Mao-jun CAI~3 Jing-kang SHEN~3 Yi-ping WANG~(2,5)2 State Key Laboratory of Drug Research.3 Department of Medicinal Chemistry,shanghai Institute of Materia Medica,shanghai Institutes for Biological sciences.chinese academy of sciences.shanghai 201203.china 4 graduate school of the chinese academy of sciences.chinese academy of sciences,shanghai 201203.china State Key Laboratory of Drug Research Chinese Academy of Sciences Shanghai China graduate school of the chinese academy of sciences Chinese Academy of Sciences Shanghai China Department of Medicinal Chemistry Shanghai Institute of Materia Medica Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai China

Aim:To express and purify lipoprotein-associated phospholipase A2（Lp-PLA2）,and to establish a screening model for Lp-PLA2inhibitors using the expressed Lp-PLA2,Methods:We cloned the full-length cDNA of Lp-PLA2 from differentiatedTHP-I cells,and subcloned the cDNA into the baculovirus transfer *** addition,we introduced an N-terminal Kozak sequence for high-level translation initiation and a C-terminal sequence of 6 histidine residues forpurification,The fusion enzyme was expressed in Sf9 insect cells and purified byNi2+affinity *** Lp-PLA2activity was measured us-ing[3H]PAF as a substrate,and we examined the enzyme activity of recombinantLp-PLA2pretreated with the known Lp-PLA2inhibitor ***:Wesuccessfully cloned the full-length Lp-PLA2gene from differentiated THP-1 *** fusion enzyme was expressed in Sf9 insect cells at a high level and purifiedefficiently through a 2-step *** recombinant Lp-PLA2activity wasmeasured using[3H]PAF as a substrate,and proved to be enzymatically ***-PLA2inhibitor SB435495 produced a good inhibition curve for inhibition ofrecombinant Lp-PLA2with an IC50of 57±1μmol/***:We expressed andpurified Lp-PLA2at a high level in insect cell-baculovirus expression *** ratio was much greater than that obtained from human plasma and we estab-lished a screening model for Lp-PLA2inhibitors using the expressed Lp-PLA2.

关键词： lipoprotein-associated phospholipase A2 atherosclerosis coronary heart diseases cloning baculovirus purification

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chinese expert consensus on immunotherapy for advanced non-small cell lung cancer with oncogenic driver mutations(2023 edition)

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chinese Medical Journal 2024年第9期137卷 1016-1018页

作者： The Society of Cancer Precision Medicine of chinese Anti-Cancer Association The Lung Cancer Expert Group of chinese Medical Association Publishing House Baohui Han Hua Zhong 不详 Department of Respiratory Medicine Shanghai Chest HospitalSchool of MedicineShanghai JiaoTong UniversityShanghai 20030China shanghai Chest Hospital Shanghai Jiao Tong University School of Medicine

Non-small cell lung cancer(NSCLC)with oncogenic driver mutations was previously deemed“forbidden territory”for immunotherapy.[1]With the continuous generation of evidence,immunotherapy is expected to bring hope for such NSCLC with oncogenic driver mutations after acquired tyrosine kinase inhibitor(TKI)***,given the intricate interplay between driven mutations,targeted therapy,and immune microenvironment,the efcacy of immunotherapy in advanced NSCLC patients with diverse oncogenic driver mutations remains a topic of ongoing *** this context,chinese Expert Consensus on Immunotherapy for Advanced NSCLC with Oncogenic Driver Mutations(2022 edition)wasrst launched in 2022 by the Society of Cancer Precision Medicine of chinese Anti-Cancer Association and the Lung Cancer Expert Group of chinese Medical Journals,aiming to provide standardized treatment guidance for clinical practice.

关键词： immunotherapy cancer lung

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2023 chinese national clinical practice guideline on diagnosis and management of Crohn’s disease

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中华医学杂志(英文版) 2024年第14期137卷 2024 Jul 2页

作者： Inflammatory Bowel Disease Group, chinese Society of Gastroenterology, chinese Medical Association Inflammatory Bowel Disease Quality Control Center of china Xijing Hospital Air Force Medical University China Peking Union Medical College Hospital China West china Hospital Sichuan University China The First Affiliated Hospital Dalian Medical University China University of Nottingham Ningbo China GRADE Centre United Kingdom The First Affiliated Hospital Sun Yat-sen University China Sir Run Run Shaw Hospital Zhejiang University School of Medicine China Peking University People's Hospital China The Second Affiliated Hospital Zhejiang University School of Medicine China Renmin Hospital Wuhan University China chinese PLA General Hospital China The First Affiliated Hospital Naval Medical University China The Sixth Affiliated Hospital Sun Yat-sen University China The First Affiliated Hospital Anhui Medical University China Zhongshan Hospital Xiamen University China Zhongnan Hospital Wuhan University China The First Hospital of Shanxi Medical University China The Eighth Affiliated Hospital Sun Yat-sen University China Peking University Third Hospital China Peking University First Hospital China Qilu Hospital Shandong University China General Hospital of Eastern Theater Command China Xiangya Hospital Central South University China The Tenth People's Hospital Tongji University China The First Affiliated Hospital Kunming Medical University China shanghai Pudong New District Zhoupu Hospital China Ruijin Hospital Shanghai Jiao Tong University School of Medicine China Tongji Medical College Huazhong University of Science and Technology China General Hospital of Nanjing Military Command China The Third Xiangya Hospital Central South University China Daping Hospital Chongqing China The Second Xiangya Hospital Central South University China Jiangsu Provincial People's Hospital China The Second Hospital of Hebei Medical University China Nanjing Drum Tower Hospital China Nanfan Hospital Southern Medical University China GRADE Center University of Nottingham Ningbo China The Fifth Affiliated Hospital Zhengzhou University

The role of chinese expert consensus in standardization and improvement of the clinical diagnosis and treatment of inflammatory bowel disease (IBD) is self-evident. As clinical and basic research in china becomes standardized under a better understanding of IBD, more data on chinese patients have become available to develop a consensus and guideline. This guideline, collaboratively developed by the IBD Group, chinese Society of Gastroenterology, incorporates the latest international consensus statements,[1-7] domestic research findings, and practical considerations, as an update based on the 2018 chinese consensus on IBD management.[8] The IBD guideline consists of two parts: ulcerative colitis (UC) and Crohn's disease, and this manuscript presented the UC part. The evidence used in this guideline was collected and analyzed by the standard of guideline, and the contents were organized as problem statements, for clarity. The formulation of this guideline aimed to reflect the latest progress in IBD, providing comprehensive and valuable guidance for the clinical management of IBD.

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Clinical practice guidelines for nutritional assessment and monitoring of adult ICU patients in china

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Journal of Intensive Medicine 2024年第2期4卷 137-159页

作者： Xiangdong Guan Dechang Chen Yuan Xu chinese Society of Critical Care Medicine Department of Critical Care Medicine The First Affiliated Hospital of Sun Yat-sen UniversityGuangzhouChina Department of Critical Care Medicine Ruijin HospitalShanghai Jiao Tong University School of MedicineShanghaiChina Department of Critical Care Medicine Beijing Tsinghua Changgung HospitalBeijingChina 不详

The chinese Society of Critical Care Medicine(CSCCM)has developed clinical practice guidelines for nutrition assessment and monitoring for patients in adult intensive care units(ICUs)in *** guideline focuses on nutrition evaluation and metabolic monitoring to achieve optimal and personalized nutrition therapy for critically ill *** guideline was developed by experts in critical care medicine and evidence-based medicine methodology and was developed after a thorough review of the system and a summary of relevant trials or studies published from 2000 to July 2023.A total of 18 recommendations were formed and consensus was reached through discussions and reviews by expert groups in critical care medicine,parenteral and enteral nutrition,and *** recommendations are based on currently available evidence and cover several key fields,including screening and assessment,evaluation and assessment of enteral feeding intolerance,metabolic and nutritional measurement and monitoring during nutrition therapy,and organ function evaluation related to nutrition *** question was analyzed according to the Population,Intervention,Comparison,and Outcome(PICO)*** addition,interpretations were provided for four questions that did not reach a consensus but may have potential clinical and research *** plan is to update this nutrition assessment and monitoring guideline using the international guideline update method within 3–5 years.

关键词： Adults Critical illness Intensive care unit Nutriti on Nutriti on assessment and monitoring Guideline Recommendations chinese Society of Critical Care Medicine-chinese Medical Association(CSCCM-CMA)

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Cloning and Analyzing of Xenopus Mespo Promoter in Retinoic Acid Regulated Mespo Expression

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Acta Biochimica et Biophysica Sinica 2006年第11期38卷 759-764页

作者： Jin-Hu WANG Xiao-Yan DING* 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 and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Graduate School of the Chinese Academy of SciencesShanghai 200031 China

During vertebrate embryogenesis,presomitic mesoderm cells enter a segmental program togenerate somite,a process termed ***,a member of the bHLH transcription factor family,plays important roles in this ***,how Mespo expression is regulated remains *** this question,we isolated a genomic DNA sequence containing 4317 bp of Mespo 5’ flanking regionin *** assays show that this upstream sequence has transcription *** assayshows that this genomic contig is sufficient to recapitulate the dynamic stage-and tissue-specific expressionpattern of endogenous Mespo from the gastrula to the tailbud *** further mapped a 326 bp DNAsequence responding to retinoic acid *** results shed light on how Mespo expression is regulated,and suggest that retinoic acid signaling pathways play roles in somitogenesis through regulating Mespo.

关键词： bHLH paraxial presomitic segmental tailbud Xenopus laevis

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Comparative studies of huperzine A,donepezil,and rivastigmine on brain acetylcholine,dopamine,norepinephrine,and 5-hydroxytryptamine lev-els in freely-moving rats

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Acta Pharmacologica Sinica 2006年第9期27卷 1127-1136页

作者： Yan-qi LIANG Xi-can TANG~2 State Key Laboratory of Drug Research,shanghai Institute of Materia Medica,shanghai Instituties for Biological sciences,graduate school of the chinese academy of sciences.chinese academy of sciences,shanghai 201203.china State Key Laboratory of Drug Research Shanghai Institute of Materia Medica Shanghai Instituties for Biological Sciences Graduate School of the Chinese Academy of Sciences Chinese Academy of Sciences Shanghai China

Aim:To assess the effects of cholinesterase inhibitors huperzine A,donepeziland rivastigmine on cerebral neurotransmitters in the cortex and hippocampus infreely-moving ***:Double-probe cerebral microdialysis and HPLC withelectrochemical detection were used to detect ***:Ourresults showed that huperzine A(0.***.0.5,and 0.75μmol/kg,po)dose-depen-dently elevated extracellular acetylcholine(ACh)levels in the medial prefrontalcortex(mPFC)and *** administration of donepezil(5.4μmol/kg)orrivastigmine(1μmol/kg)also elicited significant increases in ACh in the mPFC *** time course of cortical acetylcholinesterase(ACHE)inhibitionwith the 3 inhibitors minored the increases of ACh at the same *** markedelevation of ACh after oral administration of huperzine A(0.5μmol/kg)anddonepezil(5.4μmol/kg)was associated with a significantly increased release ofdopamine(DA)in the mPFC or *** of the 3 inhibitors affectednorepinephrine(NE)and 5-hydroxytryptamine(5-HT)levels in the mPFC *** effects of huperzine A and rivastigmine did not depend on theroute of administration,but donepezil was less efficacious by the oral route thanby ip *** ability of huperzine A to increase ACh levels was unchangedwhen tests were performed after multiple oral administration of the drug at 0.5μmol/kg,once per day for 30 ***:The present findings showed that,inmolar terms,huperzine A had similar potency on increasing mPFC ACh and DAlevels as compared to the 11-and 2-fold dosages of donepezil and rivastigmine,respectively,and had longer lasting effects after oral dosing.

关键词： huperzine A donepezil rivastigmine acetylcholine dopamine norepinephrine 5-hydroxy tryptamine Alzheimer’s disease rats

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Development of a complex scintillation proximity assay for highthroughput screening of PPAR7 modulators

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Acta Pharmacologica Sinica 2005年第3期26卷 339-344页

作者： Bin WU Jie GAO Ming-wei WANG~2 The National Center for Drug Screening,shanghai Institute of Materia Medica,shanghai Institutes for Biological sciences.chinese academy of sciences graduate school of chinese academy of sciences,shanghai 201203.china The National Center for Drug Screening Shanghai Institute of Materia Medica Shanghai Institutes for Biological Sciences Chinese Academy of Sciences graduate school of chinese academy of sciences Shanghai China

Aim:To develop a complex high-throughput screening （HTS） assay based onscintillation proximity assay （SPA） technology for identification of novel peroxi-some proliferator-activated receptor gamma （PPARγ） ***:Full-length PPARγ and retinoid X receptor alpha （RXRα）,biotinylated PPAR responseelement （PPRE）,[3H]BRL49653 and streptavidin-coated FlashPlate or microbeadwere used to develop an HTS assay based on SPA ***‘ABCDE’method was validated against conventional hydroxyapatite （HA） assay and ap-plied to large-scale screening of 16 000 synthetic compounds and natural ***:（1） IC50values of positive control compounds （BRL49653 andtroglitazone） obtained from the‘ABCDE’method and HA assay were compa-rable and consistent with those reported elsewhere;（2） Approximately 178compounds,showing more than 70% competitive inhibition on BRL49653 bind-ing to PPARγ,were identified initially by the‘ABCDE’method （microbead）;（3）Secondary screening using FlashPlate and cross-reactivity studies with RARα,β,γ and RXRα,β,γ confirmed that 12 compounds possessed specific PPARγ bind-ing properties including 2 with IC50values less than 0.5 μmol/L and novel chemi-cal ***:The‘ABCDE’method using either FlashPlate ormicrobead,is a highly efficient,automatable,and robust tool to screen potentialPPARγ modulators in HTS *** application may be expanded to othernuclear receptors that form heterodimers upon activation.

关键词： peroxisome proliferator-activated receptor gamma retinoid X receptor alpha scintillation proximity assay high-throughput screening

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In Vitro Reassembly of Tobacco Ribulose-1,5-bisphosphate Carboxylase/Oxygenase from Fully Denatured Subunits

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Acta Biochimica et Biophysica Sinica 2006年第10期38卷 737-745页

作者： Zhen-Hua YONG Gen-Yun CHEN* Jiao-Nai SHI Da-Quan XU Institute of Plant Physiology and Ecology,shanghai Institutes for Biological sciences.chinese academy of sciences,and graduate school of the chinese academy of sciences,shanghai 200032,china Institute of Plant Physiology and Ecology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences and Graduate School of the Chinese Academy of SciencesShanghai 200032 China

It has been generally proved impossible to reassemble ribulose-1,5-bisphosphate carboxylase/oxygenase （Rubisco） from fully denatured subunits in vitro in higher plant,because large subunit of fullydenatured Rubisco is liable to precipitate when the denaturant is removed by common methods of directdilution and one-step *** our experiment,the problem of precipitation was resolved by an improvedgradual dialysis method,which gradually decreased the concentration of ***,fully denaturedRubisco subunits still could not be reassembled into holoenzyme using gradual dialysis unless chaperonin 60was *** restored activity of reassembled Rubisco was approximately 8% of natural *** of reassembled Rubisco increased greatly when heat shock protein 70 was present in the *** and Mg2+ were unnecessary for in vitro reassembly of Rubisco,and Mg2+ inhibited the *** reassembly was weakened when ATP,Mg2+ and K+ existed together in the reassembly process.

关键词： Rubisco reassembly gradual dialysis chaperonin 60 Hsp70

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The cDNA and Genomic DNA Organization of a Novel Toxin SHT-I from Spider Ornithoctonus huwena

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Acta Biochimica et Biophysica Sinica 2004年第10期36卷 656-660页

作者： Peng QIAO, Xiao-Pan ZUO, Zhi-Fang CHAI, and Yong-Hua JI* Institute of Physiology, shanghai Institutes for Biological sciences, chinese academy of sciences, graduate school of the chinese academy of sciences, shanghai 200031, china Institute of Physiology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Graduate School of the Chinese Academy of SciencesShanghai 200031 China

In present study the full-length cDNA of a novel toxin from the venom gland of spider Ornithoctonus huwena, named as SHT-I, has been cloned using the strategy of rapid amplification of cDNA ends, and then the whole genomic sequence of SHT-I (Selenocosmia huwena toxin-I) was determined using sequence-specific primers synthesized based on the acquired 3’ and 5’ ends of SHT-I cDNA sequence. It is unexpectedly found that intron was lacking in the genomic sequence of SHT-I. The result might evoke an interesting question whether the gene code of inhibitor cystine-knot peptides from spider venom is distinct from that of those known toxic peptides of scorpion and cone snail.

关键词： Ornithoctonus huwena spider cDNA genomic DNA peptide toxin ICK motif

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Hyaluronan Promotes Tumor Lymphangiogenesis and Intralymphantic Tumor Growth in Xenografts

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Acta Biochimica et Biophysica Sinica 2005年第9期37卷 601-606页

作者： Li-Xia GUO Ke ZOU Ji-Hang JW Hong XIE Laboratory of Biotherapy,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 Laboratory of Biotherapy Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Graduate School of the Chinese Academy of SciencesShanghai 200031 China

Hyaluronan (HA),a high molecular weight glycosaminoglycan in the extracellular matrix,hasbeen implicated in the promotion of malignant phenotypes,including tumor ***,little isknown about the effect of HA on tumor-associated *** this study,mouse hepatocellularcarcinoma Hca-F cells combined with or without HA were injected subcutaneously into C3H/Hej mice,thenangiogenesis and lymphangiogenesis of implanted tumors were examined by immunostaining for platelet-endothelial cell adhesion molecule-1 and lymphatic vascular endothelial hyaluronan receptor-1 ***,we found HA promotes tumor lymphangiogenesis and the occurrence of intratumoral lymphaticvessels,but has little effect on tumor ***,HA also promotes intralymphatic tumorgrowth,although it is not sufficient to potentiate lymphatic *** results suggest that HA,which is elevated in most malignant tumor stroma,may also play a role in tumor progression by promotinglymphangiogenesis.

关键词： hyaluronan tumor lymphangiogenesis intralymphatic tumor growth

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