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Connective tissue growth factor differentially binds to members of the cystine knot superfamily and potentiates platelet-derived growth factor-B signaling in rabbit corneal fibroblast cells

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World Journal of Biological Chemistry 2015年第4期6卷 379-388页

作者： liya pi Pei-Yu Chung Sriniwas Sriram Masmudur M Rahman Wen-Yuan Song Edward W Scott Bryon E Petersen Gregory S Schultz Department of Pediatrics University of Florida Johnson and Johnson Vision Care Department of Ophthalmology Schepens Eye Research InstituteHarvard Medical School Department of Molecular Genetics and Microbiology University of Florida Department of Plant Pathology University of Florida Department of Obstetrics and Gynecology University of Florida

AIM:To study the binding of connective tissue growth factor(CTGF) to cystine knot-containing ligands and how this impacts platelet-derived growth factor(PDGF)-B signaling. METHODS:The binding strengths of CTGF to cystine knot-containing growth factors including vascular en-dothelial growth factor(VEGF)-A,PDGF-B,bone morphogenetic protein(BMP)-4,and transforming growth factor(TGF)-β1 were compared using the LexA-based yeast two-hybrid system. EYG48 reporter strain that carried a wild-type LEU2 gene under the control of Lex A operators and a lac Z reporter plasmid(p80plac Z) containing eight high affinity Lex A binding sites were used in the yeast two-hybrid analysis. Interactions between CTGF and the tested growth factors were evaluated based on growth of transformed yeast cells on selective media and colorimetric detection in a liquid β-galactosidase activity assay. Dissociation constants of CTGF to VEGF-A isoform 165 or PDGF-BB homo-dimer were measured in surface plasma resonance(SPR) analysis. CTGF regulation in PDGF-B presentation to the PDGF receptor β(PDGFRβ) was also quantitatively assessed by the SPR analysis. Combinational effects of CTGF protein and PDGF-BB on activation of PDGFRβ and downstream signaling molecules ERK1/2 and AKT were assessed in rabbit corneal fibroblast cells by Western analysis. RESULTS:In the LexA-based yeast two-hybrid system,cystine knot motifs of tested growth factors were fused to the activation domain of the transcriptional factor GAL4 while CTGF was fused to the DNA binding domain of the bacterial repressor protein Lex A. Yeast cotransformants containing corresponding fusion proteins for CTGF and all four tested cystine knot motifs survived on selective medium containing galactose and raffinose but lacking histidine,tryptophan,and uracil. In liquid β-galactosidase assays,CTGF expressing cells that were co-transformed with the cystine knot of VEGF-A had the highest activity,at 29.88 ± 0.91 fold above controls(P < 0.01). Cells conta

关键词： Connective tissue growth factor Vascular endotheli

Pyridoxine regulates hair follicle development via the pi3K/Akt, Wnt and Notch signalling pathways in rex rabbits

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Animal Nutrition 2021年第4期7卷 1162-1172页

作者： Gongyan Liu Guangmin Cheng Yongcui Zhang Shuxia Gao Haitao Sun liya Bai Shu Li Yanli Zhu Chunyang Wang Fuchang Li College of Animal Science and Technology Shandong Agricultural UniversityShandong Provincial Key Laboratory of Animal Biotechnology and Disease Control and PreventionTai'an 271018China Institute of Animal Husbandry and Veterinary Shandong Academy of Agricultural SciencesShandong Key Laboratory of Animal Disease Control and BreedingJinan 251000China Shandong Vocational Animal Science and Veterinary College Weifang 261061China

This study was conducted to evaluate the effect of pyridoxine on the development of hair follicles in Rex rabbits and the underlying molecular *** hundred 3-month-old Rex rabbits were randomly divided into 5 groups and fed diets supplemented with 0,5,10,20,or 40 mg/kg *** hair follicle density on the dorsal skin and the gene and protein expression levels of components of the phosphoinositide 3-kinase(pi3 K)/protein kinase B(PKB or Akt),Wnt,Notch and bone morphogenetic protein(BMP)signalling pathways were *** addition,free hair follicles were isolated from Rex rabbits and cultured with pyridoxine in vitro to measure hair shaft ***,dermal papilla cells(DPC)were isolated from the skin of Rex rabbits and cultured with pyridoxine in vitro to measure the gene and protein expression levels of components of the pi3 K/Akt,Wnt,Notch and BMP signalling *** results showed that the addition of dietary pyridoxine significantly increased the total follicle density,secondary follicle density,and secondary-to-primary ratio(S/P,Ppi3 K/Akt,Wnt and Notch signalling pathways in the skin and DPC of Rex rabbits(Ppi3 K/Akt,Wnt and Notch signa

关键词： Hair follicle development Pyridoxine Rex rabbit Dermal papilla cell

胶质细胞源性神经营养因子/pi3K/AKT通路参与电针促进面神经压榨损伤模型兔的面神经再生

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中国组织工程研究 2020年第7期24卷 1094-1100页

作者：费静郑红弟余莉亚李雷激西南医科大学附属医院耳鼻咽喉头颈外科

背景:多穴位电针治疗周围性面瘫的机制不明。胶质细胞源性神经营养因子(glial cell-derived neurotrophic factor,GDNF)是促进体外运动神经元存活的最有效因子,pi3K/AKT信号通路在保护受损神经元方面发挥着重要作用。目前暂没有GDNF/pi3... 详细信息

背景:多穴位电针治疗周围性面瘫的机制不明。胶质细胞源性神经营养因子(glial cell-derived neurotrophic factor,GDNF)是促进体外运动神经元存活的最有效因子,pi3K/AKT信号通路在保护受损神经元方面发挥着重要作用。目前暂没有GDNF/pi3K/AKT通路参与电针促进兔周围面神经再生的研究。目的:观察电针对周围面神经压榨损伤后再生的影响,并从GDNF/pi3K/AKT介导的信号通路角度探讨电针对面神经元的保护机制。方法:成年健康新西兰大白兔由西南医科大学动物实验中心提供,随机分成正常组和模型组。模型组制备右侧面神经颊支压榨性损伤的病理模型,造模成功后随机分为模型对照组和电针组,模型对照组自然康复,电针组取右侧翳风、颊车、四白、地仓、阳白、颧髎穴位进行电针治疗,1次/d,每次30 min。观察动物面瘫症状的改善情况并进行评分;正常组直接取材,模型组分别于术后1,4,7,14,28 d取含面神经元的脑桥组织,进行苏木精-伊红染色观察神经元形态,尼氏染色观察尼氏小体,免疫组化测定GDNF的阳性表达,Western blotting检测3组各时间点面神经元组织中GDNF、pi3K、AKT、P-AKT的蛋白表达。实验方案经西南医科大学动物伦理学委员会批准,批准号为20170120001。结果与结论:①与模型对照组相比,电针组动物的面瘫症状(患侧口角歪斜下垂,触须倒伏且运动减弱,眼睑上抬不能)恢复较快且完全;②电针组面神经元形态学改变、尼氏体变化均较模型对照组病理变化轻;③术后各时间点,面神经元GDNF免疫反应较强,阳性细胞数多于模型对照组(除术后1 d,其余各时间点Ppi3K和P-AKT蛋白表达明显增高(Ppi3K/AKT信号通路有关。

关键词：中医学电针穴胶质细胞源性神经营养因子蛋白激酶B AKT 运动神经元面神经麻痹神经再生信号转导

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基于MEMS机构装配的微夹持器研究

中国机械工程 2014年第5期25卷 596-601页

作者：郝永平董福禄张嘉易包丽雅沈阳理工大学辽宁省先进制造与装备重点实验室沈阳110159 中国空空导弹研究院洛阳471009

微夹持器作为末端执行装置,直接决定了微装配的效率。MEMS机构中包含许多微小的活动部件和功能元件,为实现这些微小器件的稳定夹取和自动装配,设计了一种采用压电陶瓷驱动、基于柔性铰链的二级放大微夹持器结构。对该微夹持器的节点应... 详细信息

微夹持器作为末端执行装置,直接决定了微装配的效率。MEMS机构中包含许多微小的活动部件和功能元件,为实现这些微小器件的稳定夹取和自动装配,设计了一种采用压电陶瓷驱动、基于柔性铰链的二级放大微夹持器结构。对该微夹持器的节点应力、刚度及最大张合量等进行了分析计算,并对微夹持器进行了试制。实验与分析结果表明,该夹持器最大张合量是245μm,放大倍数约为12.3倍,满足MEMS机构的装配要求。在此基础上,重点对张合量与夹持力进行了系统测试,通过对测试数据的非线性回归,推导出99.99%可靠度的驱动电压计算公式,实现了微夹持的精确控制。

关键词：微机电系统微装配微夹持器压电陶瓷柔性铰链非线性回归

沙美特罗替卡松吸入剂微生物限度检测方法研究

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发酵科技通讯 2019年第2期48卷 69-72,105页

作者：徐利水吴耿蒋次军刘大为皮晶汤丽雅海正药业(杭州)有限公司浙江杭州311404 浙江奕格资产管理有限公司浙江杭州310051

主要介绍沙美特罗替卡松吸入剂USP(美国药典)的微生物限度检测方法,包括计数法和特定微生物检查法。因该品难溶于水,且有一定的抑菌性,无法用传统的微生物检测方法进行计数法检测。通过常规法、薄膜过滤法和薄膜过滤法加中和剂法等3种... 详细信息

主要介绍沙美特罗替卡松吸入剂USP(美国药典)的微生物限度检测方法,包括计数法和特定微生物检查法。因该品难溶于水,且有一定的抑菌性,无法用传统的微生物检测方法进行计数法检测。通过常规法、薄膜过滤法和薄膜过滤法加中和剂法等3种方法的研究比较,得出通过薄膜过滤法加中和剂的方法,5个菌在试验组及稀释剂对照组中的回收率均超过标准50%的结果。确定了薄膜过滤加中和剂法适用于沙美特罗替卡松吸入剂微生物计数法检测,并提供了沙美特罗氟替卡松的有效微生物限度检测方法。

关键词：沙美特罗替卡松抑菌性微生物限度检测美国药典

Association Study of Serotonin Transporter SLC6A4 Gene in Primary Insomnia

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Association Study of Serotonin Transporter SLC6A4 Gene in Pr...

中国睡眠研究会第七届西部睡眠医学大会

作者： Wenjuan Chen Yuping Xie liya Zhou Jing Wang Xubin Wang Xiaoyan Su Peilin Hui Wei Ma Jinfeng Wang Bin Guo Jun Yang Jie Fan Yuan Zhao Shangli Zhang Yu Liu Gansu University of Traditional Chinese Medicine Sleep Medicine Center Gansu Provincial Hospital Electroencephalogram room Gansu Provincial Hospital School of Basic Medical Sciences Lanzhou University

The function of 5-hydroxytryptamine（5-HT or serotonin） is to promote arousal,inhibit slow wave sleep（SWS） and rapid-eye-movement sleep（REMS）.The solute carrier family 6 member 4（SLC6 A4） gene encodes 5-serotonin transporter（SERT）,which uptakes 5-HT from the brain synapses into the presynaptic ***,it is necessary to explore the association between SLC6 A4 gene polymorphisms and primary insomnia（pi）.A total of 129 patients with pi and 149 healthy controls completed the sleep scales,and underwent polysomnography（PSG）.The association of single-nucleotide polymorphisms（SNPs） in SLC6 A4 gene with pi was observed in all the *** SNPs were genotyped by SequenomMassArray *** were different degrees of linkage disequilibrium among 4 loci on SERT *** was a complete linkage disequilibrium between rs140700 and rs6352（D’=1.000）.A strong linkage disequilibrium was observed between rs2020936 and rs3794808（D’=0.771）,rs3794808 and rs6352（D’=0.993）,rs2020936 and rs140700（D’=0.803）,rs2020936 and rs6352（D’=0.958）,and rs140700 and rs3794808（D’=0.958）.Thehaplotype CGTT showed significant differences between pi group and control group（p=0.027）.All SNPs were distributed in both the groups ***,the probability of pi was higher when the genotypes in the four sites were arranged in a combination of CGTT.

关键词： Association study Serotonin transporter SLC6A4 gene Primary insomnia Genotypes

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γ-亚麻酸对血管性认知障碍的作用及其机制

解剖学杂志 2024年第5期47卷 420-424页

作者：石剑贡丽雅王燕河北医科大学第二医院营养科石家庄050000

目的:探究γ-亚麻酸(GLA)对血管性认知障碍(VCI)的作用及机制。方法:将30只雄性C57BL6J小鼠平均分为对照组、VCI组以及GLA组,对VCI组和GLA组行双侧颈动脉结扎建立VCI小鼠模型,对照组进行假手术。VCI组和假手术组每日以1 mg/kg生理盐水灌... 详细信息

目的:探究γ-亚麻酸(GLA)对血管性认知障碍(VCI)的作用及机制。方法:将30只雄性C57BL6J小鼠平均分为对照组、VCI组以及GLA组,对VCI组和GLA组行双侧颈动脉结扎建立VCI小鼠模型,对照组进行假手术。VCI组和假手术组每日以1 mg/kg生理盐水灌胃,GLA组每日以1 mg/kg GLA灌胃。14 d后,以Morris水迷宫检测小鼠认知障碍水平,H-E染色观察小鼠脑组织情况,以透射电镜观察海马体组织神经元情况,以免疫印迹检测小鼠pi3K/AKT/mTOR通路相关蛋白、细胞自噬相关蛋白以及神经元凋亡相关蛋白表达水平。结果:VCI组小鼠的逃逸潜伏期较GLA组和对照组长,穿越平台次数和目标象限停留时间短于其余2组;GLA组脑组织神经元形态优于VCI组,但较对照组差;VCI组pi3K/AKT/mTOR通路相关蛋白、细胞自噬相关蛋白以及神经元凋亡相关蛋白表达水平高于对照组和GLA组,GLA组高于对照组。结论:GLA可通过抑制pi3K/AKT/mTOR通路介导自噬以降低神经元凋亡,以对VCI进行治疗。

关键词： γ-亚麻酸认知障碍血管性痴呆氧化应激

200例高血压病中医辨证分型与经颅多普勒超声的关系

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河北中医 2008年第9期30卷 917-918页

作者：郭利亚李彦平河北省石家庄市中医院内科河北石家庄050051

探讨高血压病中医辨证分型与经颅多普勒超声(TCD)血流参数值之间的关系。方法应用TCD检测200例中医4型高血压病患者大脑中动脉(MCA)、大脑前动脉(ACA)及大脑后动脉(PCA)的平均血流速度(Vm)和血管搏动指数(pi),并与30例正常血压者对照观... 详细信息

探讨高血压病中医辨证分型与经颅多普勒超声(TCD)血流参数值之间的关系。方法应用TCD检测200例中医4型高血压病患者大脑中动脉(MCA)、大脑前动脉(ACA)及大脑后动脉(PCA)的平均血流速度(Vm)和血管搏动指数(pi),并与30例正常血压者对照观察。结果:肝火亢盛型、阴虚阳亢型高血压病MCV、ACA的Vm均高于正常对照组(Ppi与正常对照组比较差异无统计学意义(P>0.05);阴阳两虚型的PCA、痰湿壅盛型的MCV、ACA及PCA的Vm均低于对照组(Ppi均明显高于对照组(P<0.05)。结论TCD血流检测可在一定程度上为高血压病中医辨证分型提供客观依据。

关键词：辨证分型超声心动描记术,多普勒高血压

LY3023414靶向抑制pi3K/AKT/mTOR信号转导诱导人皮肤基底细胞癌细胞凋亡

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中国中西医结合皮肤性病学杂志 2021年第2期20卷 121-125页

作者：杨丽亚刘彩玉恩施土家族苗族自治州中心医院湖北恩施445000

目的观察LY3023414对人皮肤基底细胞癌细胞增殖、凋亡及磷脂醇3-激酶/蛋白激酶B/雷帕霉素的哺乳动物靶点(pi3K/AKT/mTOR)信号通路的影响。方法CCK8方法检测人皮肤基底细胞癌细胞增殖活性,Caspase-glot-3/-9活性测定试剂盒检测Caspase-3/... 详细信息

目的观察LY3023414对人皮肤基底细胞癌细胞增殖、凋亡及磷脂醇3-激酶/蛋白激酶B/雷帕霉素的哺乳动物靶点(pi3K/AKT/mTOR)信号通路的影响。方法CCK8方法检测人皮肤基底细胞癌细胞增殖活性,Caspase-glot-3/-9活性测定试剂盒检测Caspase-3/9活性,western blot实验分析Cleaved caspase-3、Cleaved PARP、p-P85(Y458)、p-AKT(S473和T308)和p-S6K(T389)蛋白表达水平,酶联免疫吸附试验(ELISA)法检测单链DNA(ss DNA)的水平,流式细胞术检测人皮肤基底细胞癌细胞凋亡情况。结果LY3023414对人皮肤基底细胞癌细胞(TE354T、Pri can-1和Pri can-2细胞)表现出明显药物毒性作用,并将细胞停滞在G0-G1期而抑制其增殖。LY3023414诱导TE354T细胞中caspase-3/-9活性及细胞凋亡;然而,LY3023414对正常人皮肤细胞(黑素细胞、皮肤角质形成细胞和皮肤成纤维细胞)没有药物毒性和促凋亡的作用。在分子水平,LY3023414抑制TE354T细胞中pi3K/AKT/mTOR信号转导,并降低P85(Y458)、AKT(S473和T308)和S6K(T389)的磷酸化水平。另外,LY3023414对TE354T细胞的药物毒性作用明显强于其他pi3K/AKT/mTOR信号通路抑制剂(雷帕霉素、mTOR激酶抑制剂OSI-027、AKT特异性抑制剂MK-2206和渥曼青霉素)。结论LY3023414靶向作用于pi3K/AKT/mTOR信号通路抑制人皮肤基底细胞癌细胞生长,为后期临床研究提供了理论依据。

关键词： LY3023414 人皮肤基底细胞癌细胞凋亡 pi3K/AKT/mTOR信号通路