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Laser capture microdissection and genetic analysis of carbon-labeled Kupffer cells

World Journal of Gastroenterology 2009年第14期15卷 1708-1718页

作者： stephan Gehring edmond Sabo maryann e san.m.rtin elizabeth m Dickson Chao-Wen Cheng Stephen H Gregory Department of medicine Rhode Island Hospital and The Warren Alpert Medical School of Brown University Providence RI 02903 United States Department of Pathology Rhode Island Hospital and The Warren Alpert Medical School of Brown University Providence RI 02903 United States zentrum fur Kinder- und Jugendmedizin der Universitaitsmedizin der Johannes Gutenberg-Universitait 55101 Mainz Germany

AIm： To develop a method of labeling and microdissecting mouse Kupffer cells within an extraordinarily short period of time using laser capture microdissection （LCm）. meTHODS： Tissues are complex structures comprised of a heterogeneous population of interconnected cells. LCm offers a method of isolating a single cell type from specific regions of a tissue section. LCm is an essential approach used in conjunction with molecular analysis to study the functional interaction of cells in their native tissue environment. The process of labeling and acquiring cells by LCm prior to mRnA isolation.can be elaborate, thereby subjecting the RnA to considerable degradation. Kupffer cell labeling is achieved by injecting India ink intravenously, thus circumventing the need for in vitro staining. The significance of this novel approach was validated using a cholestatic liver injury model. ReSULTS： mRnA extracted from the microdissected cell population displayed marked increases in colonystimulating factor-1 receptor and Kupffer cell receptor message expression, which demonstrated Kupffer cell enrichment. Gene expression by Kupffer ceils derived from bile-duct-ligated, versus sham-operated, mice was compared. microarray analysis revealed a significant （2.5-fold, q value 〈 10） change in 493 genes. Based on this fold-change and a standardized Pubmed search, 10 genes were identified that were relevant to the ability of Kupffer cells to suppress liver injury. COnCLUSIOn; The methodology outlined herein provides an approach to isolating high quality RnA from Kupffer cells, without altering the tissue integrity.

关键词： Kupffer cells India ink Laser capturemicrodissection Bile duct ligation DnA microarray

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Debaryomyces hansenii supplementation in low fish meal diets promotes growth,modulates microbiota and enhances intestinal condition in juvenile marine fish

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Journal of Animal Science and Biotechnology 2024年第1期15卷 253-276页

作者： Ignasi sanahuja Alberto Ruiz Joana P.Firmino Felipe e.Reyes-López Juan B.Ortiz-Delgado eva Vallejos-Vidal Lluis Tort Dariel Tovar-Ramírez Isabel m.Cerezo miguel A.moriñigo carmen Sarasquete enric Gisbert Aquaculture Program Institute of Agrifood Research and Technology(IRTA)La Rapita 43540Spain Centro de Biotecnologia Acuicola Universidad de Santiago de ChileSantiagoChile Instituto de Ciencias marinas de Andalucia(ICmAn-CSIC) Avda.Republica Saharaui no 2Campus Universitario Rio San PedroPuerto RealCadiz 11510Spain nucleo de Investigaciones Aplicadas en Ciencias Veterinarias y Agronomicas Facultad de Medicina Veterinaria y AgronomiaUniversidad de Las AmericasSantiagoChile Department of Cell Biology Physiologyand ImmunologyUniversitat Autonoma de BarcelonaBarcelonaSpain Centro de Investigaciones Biologicas del noroeste SC CIBNORLa PazMexico Department of microbiology Instituto de Biotecnologia Y Desarrollo Azul(IBYDA)Faculty of SciencesUniversity of Malaga29010 MalagaSpain SCBI Bioinformatic UnitUniversity of Malaga29590 MalagaSpain

Background The development of a sustainable business model with social acceptance,makes necessary to develop new strategies to guarantee the growth,health,and well-being of farmed *** hansenii is a yeast species that can be used as a probiotic in aquaculture due to its capacity to i)promote cell proliferation and differen-tiation,ii)have immunostimulatory effects,iii)modulate gut microbiota,and/or iv)enhance the digestive *** provide inside into the effects of *** on juveniles of gilthead seabream(Sparus aurata)condition,we inte-grated the evaluation of the main key performance indicators coupled with the integrative analysis of the intestine condition,through histological and microbiota state,and its transcriptomic *** After 70 days of a nutritional trial in which a diet with low levels of fishmeal(7%)was supplemented with 1.1%of ***(17.2×10^(5) CFU),an increase of ca.12%in somatic growth was observed together with an improve-ment in feed conversion in fish fed a yeast-supplemented *** terms of intestinal condition,this probiotic modu-lated gut microbiota without affecting the intestine cell organization,whereas an increase in the staining intensity of mucins rich in.carboxylated and weakly sulphated glycoconjugates coupled with changes in the affinity for certain lectins were noted in goblet *** in microbiota were characterized by the reduction in abundance of several groups of Proteobacteria,especially those characterized as opportunistic *** microarrays-based transcrip-tomic analysis found 232 differential expressed genes in the anterior-mid intestine of ***,that were mostly related to metabolic,antioxidant,immune,and symbiotic *** Dietary administration of *** enhanced somatic growth and improved feed efficiency param-eters,results that were coupled to an improvement of intestinal condition as histochemical and transcriptomic tools *** probiotic ye

关键词： Debaryomyces hansenii Intestine condition Low fish meal diet microbiota Transcriptomics Yeast probiotic

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Hypoxia inducible factor-1α mediates protective effects of ischemic preconditioning on eCV-304 endothelial cells

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World Journal of Gastroenterology 2007年第16期13卷 2369-2373页

作者： Liu-Bin Shi Jian-Hua Huang Bao-san Han Department of Organ Transplantation Huashan Hospital Fudan University Shanghai 200040 China Institute of Integrative medicine Huashan Hospital Fudan University Shanghai 200040 China Department of General Surgery Ruijin Hospital School of Medicine Shanghai Jiaotong University Shanghai 200025 China

AIm: To investigate whether hypoxia inducible factor-1α (HIF-1α) is linked to the protective effects of ischemic preconditioning (IP) on sinusoidal endothelial cells against ischemia/reperfusion injury. meTHODS: Sinusoidal endothelial cell lines eCV-304 were cultured and divided into four groups: control group, cells were cultured in complete Dmem medium; cold anoxia/warm reoxygenation (A/R) group, cells were preserved in a 4℃ UW solution in a mixture of 95% n2 and 5% CO2 for 24 h; anoxia-preconditioning (APC) group, cells were treated with 4 cycles of short anoxia and reoxygenation before prolonged anoxia- preconditioning treatment; and anoxia-preconditioning and hypoxia inducible factor-1α (HIF-1α) inhibitor (I-HIF-1) group, cells were pretreated with 5 μm of HIF-1α inhibitor nS398 in Dmem medium before subjected to the same treatment as group APC. After the anoxia treatment, each group was reoxygenated in a mixture of 95% air and 5% CO2 incubator for 6 h. Cytoprotections were evaluated by cell viabilities from Trypan blue, lactate dehydrogenase (LDH) release rates, and intracellular cell adhesion molecule-1 (Icam-1) expressions. expressions of HIF-1α mRnA and HIF-1α protein from each group were determined by the rt-PCR method and Western blotting, respectively. ReSULTS: Ischemia preconditioning increased cell viability, and reduced LDH release and Icam-1 expressions. Ischemia preconditioning also upregulated the HIF-1α mRnA level and HIF-1α protein expression. However, all of these changes were reversed by HIF-1α inhibitor ***: Ischemia preconditioning effectively inhibited cold hypoxia/warm reoxygenation injury to endothelial cells, and the authors showed for the first time HIF-1α is causally linked to the protective effects of ischemic preconditioning on endothelial cells.

关键词： Preconditioning Anoxia/reoxygenation injury Reperfusion injury endothelial cells Hypoxia inducible factor-1α

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淫羊藿苷对抑郁症模型小胶质细胞表型转化的调控作用

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天然产物研究与开发 2024年第2期36卷 187-195页

作者：曹利华高松王笑雨王真真贺红娟李娜白明苗明三河南中医药大学中医药科学院郑州450046 仲景宛西制药股份有限公司河南省中药固体制剂技术创新中心西峡474550 河南中医药大学药学院郑州450046

探索淫羊藿苷抗抑郁的作用机制,将SPF级Km小鼠分为空白组、模型组、盐酸氟西汀组(10 mg/kg)、淫羊藿苷高(50 mg/kg)、低(25 mg/kg)剂量组。除空白组外,其余各组小鼠采用56 d慢性不可预知温和应激建立抑郁症模型。造模第1 d开始灌胃给予... 详细信息

探索淫羊藿苷抗抑郁的作用机制,将SPF级Km小鼠分为空白组、模型组、盐酸氟西汀组(10 mg/kg)、淫羊藿苷高(50 mg/kg)、低(25 mg/kg)剂量组。除空白组外,其余各组小鼠采用56 d慢性不可预知温和应激建立抑郁症模型。造模第1 d开始灌胃给予各组小鼠相应药物,连续56 d。于给药后第53~56 d,进行行为学测试。末次给药后,取材。酶联免疫吸附剂实验(enzyme linked immunosorbent assay,eLISA)检测脑组织匀浆白细胞介素6(interleukin-6,IL-6)、白细胞介素10(interleukin-10,IL-10)、五羟色胺(5-hydroxytryptamine,5-HT)、多巴胺(dopamine,DA)、去甲肾上腺素(norepinephrine,ne)水平;实时荧光定量PCR(quantitative real-time PCR,rt-PCR)检测脑组织中IL-6、IL-10、诱导性一氧化氮合酶(inducible nitric oxide synthase,inOS)、分化簇206(cluster of differentiation 206,CD206)mRnA表达;蛋白免疫印迹(Western blot)检测脑组织inOS、CD206蛋白表达。体外培养小鼠小胶质细胞(BV-2),采用细胞增殖与毒性检测试剂盒(Cell Counting Kit-8,CCK-8)法检测淫羊藿苷的细胞毒性。BV-2细胞暴露于100μg/mL脂多糖及15μg/mL、25μg/mL的淫羊藿苷24 h后,收集细胞。免疫荧光技术检测细胞inOS、CD206蛋白表达。研究结果表明,慢性不可预知温和应激致抑郁症模型制备成功,淫羊藿苷可降低抑郁症小鼠脑组织IL-6水平及IL-6、inOS mRnA表达,升高脑组织IL-10、5-HT、DA、ne水平及IL-10、CD206 mRnA表达;降低脑组织inOS蛋白,升高CD206蛋白表达;改善模型小鼠抑郁症样行为。淫羊藿苷可降低LPS刺激的BV-2细胞inOS蛋白表达,升高CD206蛋白表达。结果表明,淫羊藿苷抑制小胶质细胞m1表型转化,促进m2表型转化,从而改善模型小鼠抑郁样行为。

关键词：淫羊藿苷抑郁症小胶质细胞表型转化细胞炎症模型

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effect of bevacizumab on the expression of fibrosis-related inflammatory mediators in ARPe-19 cells

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International Journal of Ophthalmology(english edition) 2017年第3期10卷 366-371页

作者： san-Jun Chu Zhao-Hua Zhang min Wang Hai-Feng Xu Qingdao University medical College Qingdao 266071 Shandong Province China Qingdao eye Hospital Shandong Eye Institute Shandong Academy of Medical Sciences Qingdao 266071 Shandong Province China

AIm：To investigate the effect of anti-vascular epithelial growth factor（VeGF）agents on the expression of fibrosisrelated inflammatory mediators under normoxic and hypoxic conditions,and to further clarify the mechanism underlying fibrosis after anti-VeGF therapy. meTHODS：Human retinal pigment epithelial（RPe）cells were incubated under normoxic and hypoxic *** hypoxia treatment,CoCl_2 at 200μmol/L was added to the media. ARPe-19 cells were treated as following：1）control group：no treatment; 2）bevacizumab group：bevacizumab at 0.25 mg/mL was added to the media; 3）hypoxia group：CoCl_2 at 200 μmol/L was added to the media; 4）hypoxia＋bevacizumab group：CoCl_2 at 200 μmol/L and bevacizumab at 0.25 mg/mL were added to the *** expression of interleukin（IL）-1β,IL-6,IL-8 and tumor necrosis factor（TnF）-α were evaluated using real-time polymerase chain reaction（rt-PCR）and enzyme-linked immunosorbent assay（eLISA）at 6,12,24 and 48 h. ReSULTS：Both m RnA and protein levels of IL-1β,IL-6 and IL-8 were statistically significantly higher in the bevacizumab group than in the control group at each time point,and TnF-α gene and protein expression was only significantly higher only at 24 and 48h（P〈0.05）. Under hypoxic conditions,bevacizumab significantly increased the expression of IL-1β,IL-6,IL-8 and TnF-α at 6,12,24 and 48h（P〈0.05）. IL-1β,IL-8 and TnF-α peaked at 24 h and IL-6 peaked at 12 h after the bevacizumab treatment under both normoxic and hypoxic conditions. COnCLUSIOn：Treatment of ARPe-19 cells with bevacizumab can significantly increase the expression of fibrosis-related inflammatory mediators under bothnormoxic and hypoxic conditions. Inflammatory factors might be involved in the process of fibrosis after antiVeGF therapy,and the up-regulation of inflammatory factors induced by anti-VeGF drugs might promote the fibrosis process.

关键词： bevacizumab fibrosis human retinal pigment epithelial cells inflammatory mediators

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中国中三叠世富源巨胫龙(主龙型小纲,长颈龙科)的骨骼学再描述

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古脊椎动物学报 2020年第3期58卷 169-187页

作者： Torsten m.SCHeYeR 王维李淳 Feiko mIeDemA stephan n.F.SPIeKmAn 瑞士苏黎世大学古生物研究所及博物馆苏黎世CH-8006 中国科学院地质与地球物理研究所岩石圈演化国家重点实验室北京100029 中国科学院古脊椎动物与古人类研究所中国科学院脊椎动物演化与人类起源重点实验室北京100044 中国科学院生物演化与环境卓越创新中心北京100044 德国斯图加特自然历史博物馆斯图加特D-70191

过去数十年里,中国西南地区的中三叠统产出了越来越多的爬行动物化石。以海龙类、鱼龙类和鳍龙类为主的海生爬行动物,广泛分布于整个特提斯洋周边。甚至有一些物种在这古海洋的东、西两侧,即现在的中国和欧洲都有报道。最近发现的富源... 详细信息

过去数十年里,中国西南地区的中三叠统产出了越来越多的爬行动物化石。以海龙类、鱼龙类和鳍龙类为主的海生爬行动物,广泛分布于整个特提斯洋周边。甚至有一些物种在这古海洋的东、西两侧,即现在的中国和欧洲都有报道。最近发现的富源巨胫龙(macrocnemus fuyuanensis)是较为罕见的陆生物种,该种最初发现于云南中三叠统。瑞士南部的世界遗产地圣乔治山动物群中发现的一零散标本也曾被初步推断属于该物种(作为归入种:macrocnemus ***)。由于最初对富源巨胫龙模式标本的解剖学记述有限,这一归入仅基于肢骨比例而不是骨骼特征。本文对富源巨胫龙的模式标本进行了重新描述,并与已知归入标本进行了比较。研究发现,富源巨胫龙模式标本的肩带保存比从前报道的更加完整,间锁骨形态可以被明确,而这一结构在中国唯一归入标本中只是部分暴露,但在欧洲归入标本中保存完好。在区别巨胫龙属的不同种时,推测除了肢骨比例之外,间锁骨的形态特征最为重要:富源巨胫龙间锁骨的后突短且呈纺锤状,间锁骨前缘有一对棒状的突起,从前缘中间的V形凹槽处延伸而出,以此可与巴氏巨胫龙(macrocnemus bassanii)相区别。此外,进一步详细研究了富源巨胫龙模式标本的颅骨解剖特征,发现与巴氏巨胫龙完全相同。以往对巨胫龙的腭部知之甚少,保存精美和精心修理的标本为了解该部形态特征增加了重要的信息。

关键词：特提斯三叠纪拉丁期法郎组陆生爬行动物主龙型类长颈龙类

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Accurate prediction of oxygen vacancy concentration with disordered A-site cations in high-entropy perovskite oxides

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npj Computational materials 2023年第1期9卷 2053-2065页

作者： Jiyun Park Boyuan Xu Jie Pan Dawei Zhang stephan Lany Xingbo Liu Jian Luo Yue Qi School of engineering Brown UniversityProvidenceRI 02912USA Department of Physics Brown UniversityProvidenceRI 02912USA Department of Chemical engineering and materials Science Michigan State UniversityEast LansingMI 48824USA Program of materials Science and engineering University of California San DiegoLa JollaCA 92093USA materials Science Center National Renewable Energy LaboratoryGoldenCO 80401USA Department of mechan.cal and Aerospace engineering Benjamin M.Statler College of Engineering and Mineral ResourcesWest Virginia UniversityMorgantownWV 26506USA Department of nanoengineering University of California San DiegoLa JollaCA 92093USA

entropic stabilized ABO_(3) perovskite oxides promise many applications,including the two-step solar thermochemical hydrogen(STCH)*** binary and quaternary A-site mixed{A}FeO_(3) as a model system,we reveal that as more cation types,especially above four,are mixed on the A-site,the cell lattice becomes more cubic-like but the local Fe–O octahedrons are more *** comparing four different Density Functional Theory-informed statistical models with experiments,we show that the oxygen vacancy formation energies(e^(f)_(V))distribution and the vacancy interactions must be considered to predict the oxygen non-stoichiometry(δ)*** STCH applications,the e^(f)_(V) distribution,including both the average and the spread,can be optimized jointly to improveΔδ(difference ofδbetween the two-step conditions)in some hydrogen production *** model can be used to predict the range of water splitting that can be thermodynamically improved by mixing cations in{A}FeO_(3) perovskites.

关键词： perovskite vacancy oxides

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Bevacizumab modulates retinal pigment epithelial-tomesenchymal transition via regulating notch signaling

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International Journal of Ophthalmology(english edition) 2015年第2期8卷 245-249页

作者： Jing-Jing Zhang san-Jun Chu Xiao-Lei Sun Ting Zhang Wei-Yun Shi Qingdao University medical College Shandong Provincial Key Laboratory of Ophthalmology Shandong Eye Institute Shandong Academy of Medical Sciences

AIm: To investigate the effect of bevacizumab treatment on notch signaling and the induction of epithelial-of-mesenchymal transition(emT) in human retinal pigment epithelial cells(ARPe-19) in ***: In vitro cultivated ARPe-19 cells were treated with 0.25 mg/m L bevacizumab for 12, 24, and 48 *** morphology changes were observed under an inverted microscope. The expression of zonula occludens-1(ZO-1), vimentin and notch-1 intracellular domain(nICD) was examined by *** m RnA levels of ZO-1, α-SmA, notch-1, notch-2,notch-4, Dll4, Jagged-1, RBP-Jk and Hes-1 expression were evaluated with quantitative real-time polymerase chain reaction(q rt-PCR). The protein levels of α-SmA,nICD, Hes-1 and Dll-4 expression were examined with Western ***: Bevacizumab stimulation increased the expression of α-SmA and vimentin in ARPe-19 cells which changed into spindle-shaped fibroblast-like ***, the m RnA expression of Hes-1 increased and the protein expression of Hes-1 and nICD also increased, which notch signaling was activated. The m RnA expression of notch-1, Jagged-1 and RBP-Jk increased at 48 h, and while Dll4 m RnA and protein expression did not change after bevacizumab ***: Jagged-1/notch-1 signaling may play a critical role in bevacizumab-induced emT in ARPe-19 cells, which provides a novel insight into the pathogenesis of intravitreal bevacizumab-associated complication.

关键词： bevacizumab notch signaling epithelial-tomesenchymal transition retinal pigment epithelial cells

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Sustained-release genistein from nanostructured lipid carrier suppresses human lens epithelial cell growth

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International Journal of Ophthalmology(english edition) 2016年第5期9卷 643-649页

作者： Jin-Lu Liu Wen-Ji Zhang Xue-Dong Li na Yang Wei-san Pan Jun Kong Jin-Song Zhang Department of Ophthalmology the Fourth Affiliated Hospital of China Medical UniversityEye Hospital of China Medical UniversityKey Lens Research Laboratory of Liaoning Province Department of Pharmaceutics School of PharmacyShenyang Pharmaceutical University

AIm： To design and investigate the efficacy of a modified nanostructured lipid carrier loaded with genistein（Gen-nLC） to inhibit human lens epithelial cells（HLeCs） proliferation.·meTHODS： Gen-nLC was made by melt emulsification method. The morphology, particle size（PS）, zeta potentials（ZP）, encapsulation efficiency（ee） and in vitro release were characterized. The inhibition effect of nanostructured lipid carrier（nLC）, genistein（Gen） and Gen-nLC on HLeCs proliferation was evaluated by cell counting kit-8（CCK-8） assay, gene and protein expression of the proliferation marker Ki67 were evaluated with real-time quantitative polymerase chain reaction（rt-q PCR） and immunofluorescence analyses.·ReSULTS： The mean PS of Gen-nLC was 80.12±1.55 nm with a mean polydispersity index of 0.11±0.02. The mean ZP was-7.14 ±0.38 m V and the ee of Gen in the nanoparticles was 92.3% ±0.73%. Transmission electron microscopy showed that Gen-nLC displayed spherical-shaped particles covered by an outer-layer structure. In vitro release experiments demonstrated a prolonged drug release for 72 h. The CCK-8 assay results showed the nLC had no inhibitory effect on HLeCs and Gen-nLC displayed a much more prominent inhibitory effect on cellular growth compared to Gen of the same concentration. The m RnA and protein expression of Ki67 in LeCs decreased significantly in Gen-nLC group.·COnCLUSIOn： Sustained drug release by Gen-nLCs may impede HLeC growth.

关键词： posterior capsular opacification genistein nanostructured lipid carrier human lens epithelial cells

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Anti-diabetic potential of apigenin,luteolin,and baicalein via partially activating PI3K/Akt/GLUT-4 signaling pathways in insulin-resistant HepG2 cells

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Food Science and Human Wellness 2023年第6期12卷 1991-2000页

作者： Lingchao miao Haolin Zhang meng Sam Cheong Ruting Zhong Paula Garcia-Oliveira miguel A.Prieto Ka-Wing Cheng mingfu Wang Hui cao Shaoping nie Jesus Simal-Gandara Wai san Cheang Jianbo Xiao Institute of Chinese medical Sciences University of MacaoMacao 999078China Department of Analytical Chemistry and Food Science Faculty of Food Science and TechnologyUniversity of Vigo-Ourense CampusOurense E-32004Spain Institute for Advanced Study Shenzhen UniversityShenzhen 508060China School of Biological Sciences The University of Hong KongHong Kong 999077China College of Food Science and Technology Guangdong Ocean UniversityGuangdong Provincial Key Laboratory of Aquatic Product Processing and SafetyGuangdong Province Engineering Laboratory for Marine Biological ProductsGuangdong Provincial Engineering Technology Research Center of SeafoodKey Laboratory of Advanced Processing of Aquatic Product of Guangdong Higher Education InstitutionZhanjiang 524088China State Key Laboratory of Food Science and Technology China-Canada Joint Lab of Food Science and Technology(Nanchang)Nanchang UniversityNanchang 330047China International Research Center for Food nutrition and Safety Jiangsu UniversityZhenjiang 212013China

Dietary flavonoids are abundant in natural plants and possess multiple pharmacological and nutritional *** this study,apigenin,luteolin,and baicalein were chosen to evaluate their anti-diabetic effect in high-glucose and dexamethasone induced insulin-resistant(IR)HepG2 *** flavonoids improves the glucose consumption and glycogen synthesis abilities in IR-HepG2 cells via activating glucose transporter protein 4(GLUT4)and phosphor-glycogen synthase kinase(GSK-3β).These fl avonoids signifi cantly inhibited the production of reactive oxygen species(ROS)and advanced glycation end-products(AGes),which were closely related to the suppression of the phosphorylation form of nF-κB and *** expression levels of insulin receptor substrate-1(IRS-1),insulin receptor substrate-2(IRS-2)and phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt)pathway in IR-HepG2 cells were all partially activated by the fl avonoids,with variable ***,the intracellular metabolic conditions of the fl avonoids were also evaluated.

关键词： Apigenin Luteolin Baicalein Insulin-resistant HepG2 cells Signaling pathway Reactive oxygen species(ROS) Advanced glycation end-products(AGes) Glycogen synthase kinase(GSK-3β) Glucose transporter protein 4(GLUT4)

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