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Gram-negative bacteria and their lipopolysaccharides in Alzheimer’s disease:pathologic roles and therapeutic implications

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Translational Neurodegeneration 2021年第4期10卷 643-665页

作者： Hyeon soo Kim Sujin Kim Soo Jung Shin Yong Ho Park Yunkwon Nam Chae won Kim Kang won Lee Sung-Min Kim In Duk Jung Hyun Duk Yang Yeong-Min Park Minho Moon Department of Biochemistry College of MedicineKonyang UniversityDaejeon 35365Republic of Korea Research Institute for Dementia Science Konyang UniversityDaejeon 35365Republic of Korea Dandi Bioscience Inc 6th Floor of Real Company Building66Achasan-roSungdong-guSeoulRepublic of Korea Harvard Neurology Clinic 294 Gwanggyojungang-roSuji-guYongin 16943Republic of Korea Department of Immunology School of MedicineKonkuk University268ChungwondaeroChungju-siChungcheongbuk-doRepublic of Korea

Alzheimer's disease(AD)is the most serious age-related neurodegenerative disease and causes destructive and irreversible cognitive decline.Failures in the development of therapeutics targeting amyloid-β(Aβ)and tau;principal proteins inducing pathology in AD,suggest a paradigm shift towards the development of new therapeutic targets.The gram-negative bacteria and lipopolysaccharides(LPS)are attractive new targets for AD treatment.Surprisingly,an altered distribution of gram-negative bacteria and their LPS has been reported in AD patients.Moreover,gram-negative bacteria and their LPS have been shown to affect a variety of AD-related pathologies,such as Aβ homeostasis,tau pathology,neuroinflammation,and neurodegeneration.Moreover,therapeutic approaches targeting gram-negative bacteria or gram-negative bacterial molecules have significantly alleviated AD-related pathology and cognitive dysfunction.Despite multiple evidence showing that the gram-negative bacteria and their LPS play a crucial role in AD pathogenesis,the pathogenic mechanisms of gram-negative bacteria and their LPS have not been clarified.Here,we summarize the roles and pathomechanisms of gram-negative bacteria and LPS in AD.Furthermore,we discuss the possibility of using gram-negative bacteria and gram-negative bacterial molecules as novel therapeutic targets and new pathological characteristics for AD.

关键词： Alzheimer's disease Gram-negative bacteria Lipopolysaccharide Exotoxin Amyloid beta Tau

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过程参数对超临界抗溶剂法共沉淀紫杉醇-PLLA的影响(英文)

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Chinese Journal of Chemical Engineering 2012年第4期20卷 803-813页

作者：李文锋刘贵金李黎仙伍娟吕扬效江燕斌 Star Lake bioscience Co. Inc. School of Chemistry and Chemical Engineering South China University of Technology

Paclitaxel(PTX) is an effective anticancer drug with poor solubility in water.Recently,much effort has been devoted into alternative formulations of PTX for improving its aqueous solubility.In this study,PTX and poly(L-lactic acid)(PLLA) were co-precipitated by a supercritical antisolvent(SAS) process using dichloromethane(DCM) and the mixtures of DCM/ethanol(EtOH) or DCM/dimethyl sulfoxide(DMSO) as the solvent,with super-critical carbon dioxide as the antisolvent.The effects of solvent,solvent ratio,temperature,pressure,polymer con-centration and solution flow rate on particle morphology,mass median diameter(Dp50) and PTX loading were in-vestigated using single-factor method.The particle samples were characterized using X-ray diffraction(XRD),scanning electron microscopy(SEM),laser diffraction particle size analyzer and high pressure liquid chromatogra-phy(HPLC).XRD results indicate that the micronized PTX is dispersed into the PLLA matrix in an amorphous form.SEM indicates that the solvent and the solvent ratio have great effect on the particle morphologies,and particle morphology is good at the volume ratio of DCM/EtOH of 50/50.For the mixed DCM/EtOH solvent,Dp50 increases with the increase of the temperature,pressure,PLLA concentration and solution flow rate,and PTX loading in-creases with pressure.Suitable operating conditions for the experimental system are as follows:DCM/EtOH 50/50(by volume),35 ℃,10-12 MPa,PLLA concentration of 5 g·L-1 and solution flow rate of 0.5 ml·min-1.

关键词：抗溶剂过程超临界抗溶剂 L-乳酸紫杉醇共沉淀工艺参数聚合物浓度超临界二氧化碳

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Establishment of Reproducible Xenotransplantation Model of T Cell Acute Lymphoblastic Leukemia in NOD/SCID Mice

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2012年第4期32卷 511-516页

作者：王迪王娜张艳马淑燕耿哲周鹏飞周剑峰黄亮 Department of Hematology Tongji HospitalTongji Medical CollegeHuazhong University of Science and Technology Crown bioscience inc.(Beijing)

T cell acute lymphoblastic leukemia(T-ALL) is an aggressive leukemia.However the poor prognosis and low morbidity restrict further analysis of the disease.Therefore there is an increasing demand to develop animal models for identifying novel therapeutic approaches.In this study,we inoculated the anti-mouse CD122 monoclonal antibody conditioned NOD/SCID mice with the leukemia cells from 9 T-ALL patients and 1 cell line via the tail vein.Four of the 9 patients and the cell line were successfully engrafted.Flow cytometry detected high percentage of human CD45 + cells in recipient mice.Immunohistochemistry showed infiltration of human CD45 + cells in different organs.Serial transplantation was also achieved.In vivo drug treatment showed that dexamethasone could extend survival,which was consistent with clinical observation.These results demonstrated that we successfully established 5 xenotransplantation models of T-ALL in anti-mCD122 mAb conditioned NOD/SCID mice,which recapitulated the characteristics of original disease.

关键词： T cell acute lymphoblastic leukemia xenotransplantation NOD/SCID mice in vivo

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An aerosol formulation of R-salbutamol sulfate for pulmonary inhalation

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Acta Pharmaceutica Sinica B 2014年第1期4卷 79-85页

作者： Xuemei Zhang Qing Liu Junhua Hu Ling Xu Wen Tan School of bioscience and Bioengineering South China University of TechnologyGuangzhou 510006China Key-Pharma Biomedical inc. Dongguan 523000China

An aerosol formulation containing 7.5 mg of R-salbutamol sulfate was developed.The aerosol was nebulized with an air-jet nebulizer,and further assessed according to the new European Medicines Agency(EMA)guidelines.A breath simulator was used for studies of delivery rate and total amount of the active ingredient at volume of 3 mL.A next generation impactor(NGI)with a cooler was used for analysis of the particle size and in vitro lung deposition rate of the active ingredient at 51C.The anti-asthmatic efficacy of the aerosol formulation was assessed in guinea pigs with asthma evoked by intravenous injection of histamine compared with racemic salbutamol.Our results show that this aerosol formulation of R-salbutamol sulfate met all the requirements of the new EMA guidelines for nebulizer.The efficacy of a half-dose of R-salbutamol equaled that of a normal dose of racemic salbutamol.

关键词： R-salbutamol Nebulizer NGI Particle size Dose uniformity Guinea pigs Asthma

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Streamlined human antibody generation and optimization by exploiting designed immunoglobulin loci in a B cell line

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Cellular & Molecular Immunology 2021年第6期18卷 1545-1561页

作者： Hidetaka Seo Hitomi Masuda Kenjiro Asagoshi Tomoaki Uchiki Shigehisa Kawata Goh Sasaki Takashi Yabuki Shunsuke Miyai Naoki Takahashi Shu-ichi Hashimoto Atsushi Sawada Aki Takaiwa Chika Koyama Kanako Tamai Kohei Kurosawa Ke-Yi Lin Kunihiro Ohta Yukoh Nakazaki Department of Life Sciences Graduate School of Arts and SciencesThe University of TokyoTokyoJapan Research Laboratories Chiome Bioscience IncTokyoJapan

Monoclonal antibodies(mAbs)are widely utilized as therapeutic drugs for various diseases,such as cancer,autoimmune diseases,and infectious diseases.Using the avian-derived B cell line DT40,we previously developed an antibody display technology,namely,the ADLib system,which rapidly generates antigen-specific mAbs.Here,we report the development of a human version of the ADLib system and showcase the streamlined generation and optimization of functional human mAbs.Tailored libraries were first constructed by replacing endogenous immunoglobulin genes with designed human counterparts.From these libraries,clones producing full-length human IgGs against distinct antigens can be isolated,as exemplified by the selection of antagonistic mAbs.Taking advantage of avian biology,effective affinity maturation was achieved in a straightforward manner by seamless diversification of the parental clones into secondary libraries followed by single-cell sorting,quickly affording mAbs with improved affinities and functionalities.Collectively,we demonstrate that the human ADLib system could serve as an integrative platform with unique diversity for rapid de novo generation and optimization of therapeutic or diagnostic antibody leads.Furthermore,our results suggest that libraries can be constructed by introducing exogenous genes into DT40 cells,indicating that the ADLib system has the potential to be applied for the rapid and effective directed evolution and optimization of proteins in various fields beyond biomedicine.

关键词： Immunoglobulin rearrangements homologous recombination antibody therapeutics antibody engineering drug discovery

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Solubility of Paclitaxel in Mixtures of Dichloromethane and Supercritical Carbon Dioxide

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Chinese Journal of Chemical Engineering 2011年第4期19卷 558-564页

作者： LUO Nin LU Yingmei JIANG Yanbin Star Lake bioscience Co. Inc. Zhaoqing 526040 China School of Chemistry and Chemical Engineering South China University of Technology Guangzhou 5 10640 China

Phase behavior of paclitaxel in solvent mixtures of dichloromethane and supercritical carbon dioxide was investigated using a supercritical phase monitor.Cloud point pressures were determined as a function of temperature,pressure and paclitaxel content from 313.1 to 343.1K and pressures up to 33.52 MPa.The ternary mixtures exhibit a typical lower critical solution temperature behavior.When paclitaxel content increases,the single-phase region shrinks in size.Three cubic equations of state(Redlich-Kworng,Soave-Redlich-Kwong and Peng-Robinson equation of state) coupled with the van der Waals one-fluid mixing rules were selected to correlate the experimental data.The results indicate that SRK EOS coupled with two binary interaction parameters kij and lij can predict paclitaxel solubility for the best fit of experimental data.

关键词：超临界二氧化碳紫杉醇含量三元混合物二氯甲烷溶解度低临界溶液温度立方状态方程压力测定

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Novel dioxygenases for useful amino acid production

Novel dioxygenases for useful amino acid production

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The 12th Japan-China-Korea Joint Symposium on Enzyme Engineering

作者： Makoto Hibi Sakayu Shimizu Kenzo Yokozeki Jun Ogawa Research Institute for bioscience Products & Fine Chemicals Ajinomoto Co.Inc.

＜正＞Introductionα-Ketoglutarate-dependent dioxygenases are useful for production of hydroxy amino acids,some of which exert worthwhile physiological activities on the mammals,and also are components of peptide antibiotics and chiral precursors in the chemical synthesis of other compounds.We previously developed practical bioprocess system of（2S,3R,4S）-4-hydroxyisoleucine,which had remarkable anti-diabetic activity,

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Hypocrellin A-based photodynamic action induces apoptosis in A549 cells through ROS-mediated mitochondrial signaling pathway

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Acta Pharmaceutica Sinica B 2019年第2期9卷 279-293页

作者： Shanshan Qi Lingyuan Guo Shuzhen Yan Robert J.Lee Shuqin Yu Shuanglin Chen Jiangsu Province Key Laboratory for Microbes and Functional Genomics College of Life SciencesNanjing Normal University College of Pharmacy the Ohio State University Cancer Pharmacology Crown Bioscience Inc. Jiangsu Province Key Laboratory for Molecular and Medical Biotechnology College of Life SciencesNanjing Normal University

Over recent decades, many studies have reported that hypocrellin A(HA) can eliminate cancer cells with proper irradiation in several cancer cell lines. However, the precise molecular mechanism underlying its anticancer effect has not been fully defined. HA-mediated cytotoxicity and apoptosis in human lung adenocarcinoma A549 cells were evaluated after photodynamic therapy(PDT). A temporal quantitative proteomics approach by isobaric tag for relative and absolute quantitation(iTRAQ) 2 D liquid chromatography with tandem mass spectrometric(LC–MS/MS) was introduced to help clarify molecular cytotoxic mechanisms and identify candidate targets of HA-induced apoptotic cell death. Specific caspaseinhibitors were used to further elucidate the molecular pathway underlying apoptosis in PDT-treated A549 cells. Finally, down-stream apoptosis-related protein was evaluated. Apoptosis induced by HA was associated with cell shrinkage, externalization of cell membrane phosphatidylserine, DNA fragmentation,and mitochondrial disruption, which were preceded by increased intracellular reactive oxygen species(ROS) generations. Further studies showed that PDT treatment with 0.08 mmol/L HA resulted in mitochondrial disruption, pronounced release of cytochrome c, and activation of caspase-3,-9, and-7.Together, HA may be a possible therapeutic agent directed toward mitochondria and a promising photodynamic anticancer candidate for further evaluation.

关键词： Hypocrellin A Photodynamic therapy Reactive oxygen species Proteomic LC–MS/MS iTRAQ

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Thermodynamic contributions of 5’- and 3’-single strand dangling-ends to the stability of short duplex DNAs

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Journal of Biophysical Chemistry 2012年第1期3卷 1-15页

作者： Rebekah Dickman Fidelis Manyanga Greg P. Brewood Daniel J. Fish Cameron A. Fish Charlie Summers M. Todd Horne Albert S. Benight Portland bioscience Inc. Portland USA

Differential scanning calorimetry (DSC) melting analysis was performed on 27 short double stranded DNA duplexes containing 15 to 25 base pairs. Experimental duplexes were divided into two categories containing either two 5’ dangling-ends or one 5’ and one 3’ dangling-end. Duplex regions were incrementally reduced from 25 to 15 base pairs with a concurrent increase in length of dangling-ends from 1 to 10 bases. Blunt-ended duplexes from 15 to 25 base pairs served as controls. An additional set of molecules containing 21 base pair duplexes and a single four base dangling-end were also examined. DSC melting curves were measured in varying concentrations of sodium ion (Na+). From these measurements, thermodynamic parameters for 5’ and 3’ dangling-ends were evaluated as a function of dangling end length. 5’ ends were found to be slightly stabilizing but essentially constant while the 3’ ends were destabilizing with increasing length of the dangling-end. 3’ ends also display a stronger dependence on Na+ concentration. In lower Na+ environment, the 3’ ends were more destabilizing than in higher salt environment suggesting a more significant electrostatic component of the destabilizing interactions. Analysis of thermodynamic parameters of dangling ended duplexes as a function of Na+ concentration indicated the 3' dangling ends behave differently than 5' dangling ended and blunt-ended duplexes. Molecules with one 5' and one 3' dangling end showed variation in excess specific heat capacity (ΔCp) when compared to the blunt-ended molecule, while the molecules with two 5’ ends had ΔCp values that were essentially the same as blunt-ended duplexes. These observations suggested differences exist in duplexes with 3’ and 5’ dangling ends, which are interpreted in terms of composite differences in interactions with Na+, solvent, and terminal base pairs of the duplex.

关键词： DNA Thermodynamics Dangling-Ends DNA Stability

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Highly sensitive detection of melamine based on reversed phase liquid chromatography mass spectrometry

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Chinese Science Bulletin 2009年第5期54卷 732-737页

作者： WU QingQing FAN KeXin SHA Wei RUAN HongQiang ZENG Rong SHIEH ChiaHui Key Laboratory of Systems Biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China Shanghai KeyTag bioscience inc. Shanghai 200233 China

In this work, we developed a highly sensitive method to detect melamine based on reversed phase liquid chromatography mass spectrometry. A mass spectrometry compatible ion pair, heptafluorobu-tyric acid(HFBA), was used to separate melamine by reversed phase liquid chromatography prior to electrospray mass spectrometry. The incorporation of isotope internal standard and multiple reaction monitoring improved the accuracy and linearity of quantification. Based on this strategy, the method limit of quantification was 0.1 ng/g. The limits of quantification were 8 ng/g for liquid milk and 15 ng/g for dry milk powder. This method provided a reproducible and stable approach to sensitive detection and quantification of melamine.

关键词：反相流相色谱质谱分析法离子对多反应监测模式三聚氰胺

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