简介：AbstractBackground:Topoisomerase II alpha (TOP2A) has been reported to play a crucial role in the tumorigenesis of various cancer types. However, the biological role of TOP2A in gallbladder cancer (GBC) remains unknown. The current study aimed to explore the function and potential mechanism of TOP2A in GBC.Methods:Based on Gene Expression Profiling Interactive Analysis data, we found TOP2A was significantly up-regulated in GBC tissues and resulting in shorter overall survival. Quantitative real-time polymerase chain reaction and immunohistochemistry were conducted to detect the expression of TOP2A in 45 pairs of GBC tissues and adjacent non-tumor tissues. In vitro, cell proliferation, migration, and invasion ability were examined by cell counting kit-8 and transwell assay, respectively. Epithelial-mesenchymal transition (EMT) related and phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin (PI3K/Akt/mTOR) pathway-related markers were measured by Western blotting. Xenograft model assay was performed to evaluate the effect of TOP2A in vivo.Results:TOP2A was found up-regulated in GBC (tumor vs. normal, 12.62 vs. 0.34) and correlated with the late tumor node metastasis stage (P = 0.0032), present of lymph node metastasis (P = 0.0273), and poor prognosis in GBC patients (log-rank P = 0.028). In vitro and in vivo assays showed that knockdown of TOP2A notably inhibited cell proliferation, migration, invasion, EMT process, and tumor growth in GBC. In addition, TOP2A down-regulation significantly decreased the protein levels of phosphor (p)-PI3K, p-Akt, and p-mTOR.Conclusion:Our study demonstrates that TOP2A was overexpressed in GBC and associated with poor prognosis in GBC patients. TOP2A promotes GBC cell proliferation, migration, invasion, EMT process, and tumor growth through activating PI3K/Akt/mTOR signaling pathway, and may serve as a novel prognostic biomarker and therapeutic target for GBC.

简介：AbstractBackground:Vascular endothelial dysfunction is considered a key pathophysiologic process for the development of acute lung injury. In this study, we aimed at investigating the effects of unfractionated heparin (UFH) on the lipopolysaccharide (LPS)-induced changes of vascular endothelial-cadherin (VE-cadherin) and the potential underlying mechanisms.Methods:Male C57BL/6 J mice were randomized into three groups: vehicle, LPS, and LPS + UFH groups. Intraperitoneal injection of 30 mg/kg LPS was used to induce sepsis. Mice in the LPS + UFH group received subcutaneous injection of 8 U UFH 0.5 h before LPS injection. The lung tissue of the mice was collected for assessing lung injury by measuring the lung wet/dry (W/D) weight ratio and observing histological changes. Human pulmonary microvascular endothelial cells (HPMECs) were cultured and used to analyze the effects of UFH on LPS- or tumor necrosis factor-alpha (TNF-α)-induced vascular hyperpermeability, membrane expression of VE-cadherin, p120-catenin, and phosphorylated myosin light chain (p-MLC), and F-actin remodeling, and on the LPS-induced activation of the phosphatidylinositol-3 kinase (PI3K)/serine/threonine kinase (Akt)/nuclear factor kappa-B (NF-κB) signaling pathway.Results:In vivo, UFH pretreatment significantly attenuated LPS-induced pulmonary histopathological changes (neutrophil infiltration and erythrocyte effusion, alveolus pulmonis collapse, and thicker septum), decreased the lung W/D, and increased protein concentration (LPS vs. LPS + UFH: 0.57 ± 0.04 vs. 0.32 ± 0.04 mg/mL, P = 0.0092), total cell count (LPS vs. LPS + UFH: 9.57 ± 1.23 vs. 3.65 ± 0.78 × 105/mL, P= 0.0155), polymorphonuclear neutrophil percentage (LPS vs. LPS+ UFH: 88.05% ± 2.88% vs. 22.20% ± 3.92%, P = 0.0002), and TNF-α (460.33 ± 23.48 vs. 189.33 ± 14.19 pg/mL, P = 0.0006) in the bronchoalveolar lavage fluid. In vitro, UFH pre-treatment prevented the LPS-induced decrease in the membrane expression of VE-cadherin (LPS vs. LPS + UFH: 0.368 ± 0.044 vs. 0.716 ± 0.064, P = 0.0114) and p120-catenin (LPS vs. LPS + UFH: 0.208 ± 0.018 vs. 0.924 ± 0.092, P = 0.0016), and the LPS-induced increase in the expression of p-MLC (LPS vs. LPS + UFH: 0.972 ± 0.092 vs. 0.293 ± 0.025, P = 0.0021). Furthermore, UFH attenuated LPS- and TNF-α-induced hyperpermeability of HPMECs (LPS vs. LPS + UFH: 8.90 ± 0.66 vs. 15.84 ± 1.09 Ω·cm2, P = 0.0056; TNF-α vs. TNF-α + UFH: 11.28 ± 0.64 vs. 18.15 ± 0.98 Ω·cm2, P = 0.0042) and F-actin remodeling (LPS vs. LPS + UFH: 56.25 ± 1.51 vs. 39.70 ± 1.98, P = 0.0027; TNF-α vs. TNF-α + UFH: 55.42 ± 1.42 vs. 36.51 ± 1.20, P = 0.0005) in vitro. Additionally, UFH decreased the phosphorylation of Akt (LPS vs. LPS + UFH: 0.977 ± 0.081 vs. 0.466 ± 0.035, P = 0.0045) and I kappa B Kinase (IKK) (LPS vs. LPS + UFH: 1.023 ± 0.070 vs. 0.578 ± 0.044, P = 0.0060), and the nuclear translocation of NF-κB (LPS vs. LPS + UFH: 1.003 ± 0.077 vs. 0.503 ± 0.065, P = 0.0078) in HPMECs, which was similar to the effect of the PI3K inhibitor, wortmannin.Conclusions:The protective effect of UFH against LPS-induced pulmonary endothelial barrier dysfunction involves VE-cadherin stabilization and PI3K/Akt/NF-κB signaling.

简介：AbstractBackground:Autophagy of alveolar macrophages is a crucial process in ischemia/reperfusion injury-induced acute lung injury (ALI). Bone marrow-derived mesenchymal stem cells (BM-MSCs) are multipotent cells with the potential for repairing injured sites and regulating autophagy. This study was to investigate the influence of BM-MSCs on autophagy of macrophages in the oxygen-glucose deprivation/restoration (OGD/R) microenvironment and to explore the potential mechanism.Methods:We established a co-culture system of macrophages (RAW264.7) with BM-MSCs under OGD/R conditions in vitro. RAW264.7 cells were transfected with recombinant adenovirus (Ad-mCherry-GFP-LC3B) and autophagic status of RAW264.7 cells was observed under a fluorescence microscope. Autophagy-related proteins light chain 3 (LC3)-I, LC3-II, and p62 in RAW264.7 cells were detected by Western blotting. We used microarray expression analysis to identify the differently expressed genes between OGD/R treated macrophages and macrophages co-culture with BM-MSCs. We investigated the gene heme oxygenase-1 (HO-1), which is downstream of the phosphoinositide 3-kinase/protein kinase B (PI3K/Akt) signaling pathway.Results:The ratio of LC3-II/LC3-I of OGD/R treated RAW264.7 cells was increased (1.27 ± 0.20 vs. 0.44 ± 0.08, t = 6.67, P < 0.05), while the expression of p62 was decreased (0.77 ± 0.04 vs. 0.95 ± 0.10, t = 2.90, P < 0.05), and PI3K (0.40 ± 0.06 vs. 0.63 ± 0.10, t = 3.42, P < 0.05) and p-Akt/Akt ratio was also decreased (0.39 ± 0.02 vs. 0.58 ± 0.03, t = 9.13, P < 0.05). BM-MSCs reduced the LC3-II/LC3-I ratio of OGD/R treated RAW264.7 cells (0.68 ± 0.14 vs. 1.27 ± 0.20, t = 4.12, P < 0.05), up-regulated p62 expression (1.10 ± 0.20 vs. 0.77 ± 0.04, t = 2.80, P < 0.05), and up-regulated PI3K (0.54 ± 0.05 vs. 0.40 ± 0.06, t = 3.11, P < 0.05) and p-Akt/Akt ratios (0.52 ± 0.05 vs. 0.39 ± 0.02, t = 9.13, P < 0.05). A whole-genome microarray assay screened the differentially expressed gene HO-1, which is downstream of the PI3K/Akt signaling pathway, and the alteration of HO-1 mRNA and protein expression was consistent with the data on PI3K/Akt pathway.Conclusions:Our results suggest the existence of the PI3K/Akt/HO-1 signaling pathway in RAW264.7 cells under OGD/R circumstances in vitro, revealing the mechanism underlying BM-MSC-mediated regulation of autophagy and enriching the understanding of potential therapeutic targets for the treatment of ALI.

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简介：胰岛素不仅在糖尿病中发挥重要的作用，在骨的代谢中也是关键的调节因子，胰岛素与胰岛素受体结合，促进胰岛素受体底物（IRS）发生酪氨酸磷酸化，进而激活PI3kinase／Akt通路。PI3kinase／Akt通路参与了多种细胞的增殖分化，越来越多的证据证明，PI3kinase／Akt通路在骨细胞的增殖、分化起重要作用。本文就PI3kinase／Akt通路对骨代谢的调节作一综述，希望能为骨代谢疾病患者种植牙的药物选择提供思路。

简介：Phosphatidylinositolpolyphosphates(PIPx)arerelatedwithtyrosinekinaseactivation,cellproliferationandcarcinogenesis.InordertoinvestigatetheactionmechanismofPIPx,itisdesirabletosynthesizeaffinitycolumnofPI-3,4-P2,whichisexpectedtobeabletoisolatethebindingproteinsofPI-3,4-P2.ThyraminereactedwithCH-Sepharose4Bgivingcolumn13.Thep-aminogroupof3’-(1’,2’-distearoyl-glyceryl)-1-(2-p-aminobenzyl)-3,4-di-O-phosphoryl-myo-inositylphosphate(12)wasdiazotized,thendiazo-coupledwithcolumn13togivePI-3,4-P2affinitycolumn14.ThisPI-3,4-P2affinitycolumnisaneffectivetooltopickupbindingproteinsofPI-3,4-P2.

简介：hPFTAIRE1(PFTK1)，Cdc2相关的蛋白质kinase，高度在人的大脑被表示。它在Hela房间展出细胞质的分发，尽管它在它的N终点包含二个原子本地化信号(NLS)。到为它的底层和规章的部件的搜索，我们由把全身的hPFTAIRE1用作一个诱饵屏蔽了一个二混血儿的图书馆。四14-3-3isoforms(贝它，epsilon，希腊语字母的第七字，字形物)被识别与hPFTAIRE1交往。我们在hPFTAIRE1发现了一个通常认为的14-3-3绑定一致主题(RHSSPSS)，它与它的第二NLS重叠了。RHSSPSS主题的删除或有在保存有约束力的主题的翼的Ser119的替换废除了在hPFTAIRE1和14-3-3蛋白质之间的特定的相互作用。变异的S120AhPFTAIRE1也显示出一个弱相互作用到14-3-3蛋白质。结果建议Ser119为在hPFTAIRE1和14-3-3蛋白质之间的相互作用是关键的。当熔化了到绿荧光灯的蛋白质(GFP)的C终点时，所有hPFTAIRE1异种在Hela房间和人的neuroblastoma房间(SH-SY5Y)的细胞质散布了，显示有14-3-3蛋白质的那绑定不贡献潜水艇hPFTAIRE1的细胞的本地化，尽管绑定可以涉及它的发信号的规定。

简介：干扰素规章的因素(IRF)7被表明了是导致病毒的类型的一个主人管理者我干扰素生产(IFN)，并且它对病毒在天生的有免疫力的反应起一个中央作用。这里，我们识别了联系死亡的蛋白质kinase1(DAPK1)作为由双人脚踏车亲密关系纯化(龙头)的交往IRF7蛋白质。病毒的感染导致了DAPK1-IRF7和DAPK1-IRF3相互作用，DAPK1的overexpression提高了刺激干扰素的反应元素(ISRE)和IFN-β的导致病毒的激活；倡导者和IFNB1基因的表示。稀释的DAPK1击倒IFNB1和RIG-I表示的正式就职由病毒的感染或IFN-β被触发；，并且他们被病毒的复制提高。另外，病毒的感染或IFN-β；处理导致了DAPK1的表示。IFN-β；处理也由减少激活DAPK1它在丝氨酸308点的phosphorylation水平。有趣地，在导致病毒的发信号的DAPK1的参与独立于它的kinase活动。因此，我们的学习作为细胞的抗病毒的反应的一个重要管理者识别了DAPK1。

简介：AbstractPsoriatic arthritis (PsA) is a type of chronic inflammatory arthritis which is associated with psoriasis. The early recognition and treatment for PsA are of critical importance. Janus kinase (JAK) inhibitors, as a kind of orally small molecules, have emerged as an encouraging class of drug in PsA treatment. This review provides a discussion of the role and current status of JAK inhibitors in the control of PsA. There are three JAK inhibitors approved for use in autoimmune diseases, for example, tofacitinib, baricitinib, and upadacitinib, and only tofacitinib has been approved in PsA treatment. The clinical trials of upadacitinib and filgotinib in PsA patients are undergoing. The efficacy and safety of these agents were briefly discussed. Although there are still issues in terms of their efficacy and safety currently, JAK inhibitors are expected to benefit more PsA patients in future.

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简介：SulfatecanbeactivatedbyATPsulfurylaseandadenosine5-phosphosulfatekinase(APSK)invivo.RecentstudiessuggestedthatAPSKinArabidopsisthalianaregulatedthepartitionbetweenAPSreductionandphosphorylationanditsactivitycanbemodulatedbycellularredoxstatus.InordertostudyregulationofAPSKinrice(OsAPSK),OsAPSKIgenewasclonedanditsactivitywasanalyzed.OsAPSKIC36andC69werefoundtobetheconservedcounterpartsofC86andC119,whichinvolvedindisulfideformationinAtAPSK.C36A/C69AOsAPSK1doublemutationwasmadebysitedirectedmutagenesis.OsAPSKanditsmutantwereprokaryoticallyover-expressedandpurified,andthenassayedforAPSphosphorylationactivity.OsAPSK1activitywasdepressedbyoxidizedglutathione,whiletheactivityofitsmutantwasnot.Furtherstudiesinthecasethatoxidativestresswillfluctuateinvivo3'-phosphoadenosine-5'-phosphosulfatecontent,andallAPSKisoenzymeshavesimilarregulationpatternsarenecessarytobeperformed.

简介：Brassinosteroids(BR)是植物激素的一个主要的组调整植物生长和开发。BRI1，对质膜局部性的蛋白质，当BR受体和它被建议了，工作它的kinase活动在调整BR的植物生长和开发有一个必要角色。这里，我们报导隔离和bri1的新等位基因的分子的描述，bri1-301，哪个表演中等词法显型和减少的回答到在正常生长条件下面的BR。顺序分析从GG识别了二底的改变到，导致到在BRI1kinase领域的989I的989G的变换在。kinase活动的试管内试金证明bri1-301不向BRI1底层TTL和BAK1举办可检测的autophosphorylation活动或磷酸化活动。而且，我们的结果建议甚至与极其损害的kinase活动，bri1-301仍然在调整植物生长和开发保留部分功能，它提出BRI1kinase活动是否在高等植物为调停BR的生长和开发是必要的问题。

简介：Humanpolymorphonuclearleukocytes(PMN)havebeenreportedtocompletelylackofDNA-dependentproteinkinase(DNA-PK)whichiscomposedofKuproteinandthecatalyticsubunitDNA-PKcs,neededfornonhomologousend-joining(NHEJ)ofDNAdouble-strandbreaks.PromyelocyticHL-60cellsexpressavariantformofKuresultinginenhancedradiationsensitivity.ThisraisesthequestioniflowefficiencyofNHEJ,instrumentalforthecellularrepairofoxidativedamage,isanormalcharacteristicofmyeloiddifferentiation.HereweconfirmedthecompletelackofDNAPKinPMNproteinextracts,andtheexpressionofthetruncatedKu86variantforminHL-60.However,thisdegradationofDNA-PKwasshowntobeduetoaDNA-PK-degradingproteaseinPMNandHL-60.Inaddition,byusingaprotease-resistantwholecellassay,bothKu86andDNA-PKcscouldbedemonstratedinPMN,suggestingthepreviouslyreportedabsenceinPMNofDNA-PKtobeanartefact.ThelevelsofKu86andDNA-PKcsweremuchreducedinPMN,ascomparedwiththatofthelymphocytes,whereasHL-60displayedamarkedlyelevatedDNA-PKconcentration.Inconclusion,ourfindingsprovideevidenceofreduced,notdepletedexpressionofDNA-PKduringthematurestagesofmyeloiddifferentiation.

简介：SulfatecanbeactivatedbyATPsulfurylaseandadenosine5’-phosphosulfatekinase(APSK)invivo.RecentstudiessuggestedthatAPSKinArabidopsisthalianaregulatedthepartitionbetweenAPSreductionandphosphorylationanditsactivitycanbemodulatedbycellularredoxstatus.InordertostudyregulationofAPSKinrice(OsAPSK),OsAPSK1genewasclonedanditsactivitywasanalyzed.OsAPSK1C36andC69werefoundtobetheconservedcounterpartsofC86andC119,whichinvolvedindisulfideformationinAtAPSK.C36A/C69AOsAPSK1doublemutationwasmadebysitedirectedmutagenesis.OsAPSK1anditsmutantwereprokaryoticallyover-expressedandpurified,andthenassayedforAPSphosphorylationactivity.OsAPSK1activitywasdepressedbyoxidizedglutathione,whiletheactivityofitsmutantwasnot.Furtherstudiesinthecasethatoxidativestresswillfluctuateinvivo3’-phosphoadenosine-5’-phosphosulfatecontent,andallAPSKisoenzymeshavesimilarregulationpatternsarenecessarytobeperformed.

简介：1-methyl-4-phenylpyridiniumion(MPP+)inducesendoplasmicreticulumstressandactivatescaspase-12inPC12cells,leadingtoneuronalapoptosis.However,theunderlyingmolecularmechanismremainsunknown.Thepresentstudyinvestigatedtheregulatoryeffectsofnervegrowthfactor(Aktactivator)andlithiumchloride(glycogensynthasekinase-3βinhibitor)ontheendoplasmicreticulumstresssignalingpathway.TheresultsrevealedthatMPP+inducedexpressionofBipandC/EBPhomologousprotein.TheupregulationofBipandC/EBPhomologousprotein,aswellasthedecreasedpro-caspase-12levelinducedbyMPP+wereinhibitedbypretreatmentofthenervegrowthfactororlithiumchloride.Theseresultssuggestthatthephosphatidylinositol3kinase-Akt-glycogensynthasekinase-3βpathwayisinvolvedinMPP+-inducedendoplasmicreticulumstress.

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简介：房间有大量的控制维持他们的完整并且阻止随机切换到另外一个一个生物状态。RafKinase禁止的蛋白质(RKIP)，phosphatidylethanolamine绑定蛋白质(PEBP)的一个成员家庭，表明工作维持“yinyang”或生物系统的平衡的串联的调节的人的一个新类是代表性的。RKIP禁止地图kinase(Raf-MEK-ERK)，表明串联的G联合蛋白质的受体(GPCR)kinase和NFkappaB。因为RKIP指向依赖于它磷酸化的状态的不同家族ases，RKIP也行动集成多重环境刺激开始的串音。RKIP的损失或弄空导致正常细胞的壅滞和罐头的混乱导致象癌症那样的chromosomal畸形和疾病状态。因为RKIP和PEBP家庭以前被考察了，这分析的目标是提供更改并且加亮一些RKIP的唯一的特征在细胞的发信号的过程的规定使它成为一个批评播放器。

简介：Humanrhodopsinkinase(RK)andacarboxylterminus-truncatedmutantRKlackingthelast59aminoacids(RKC)wereexpressedinhumanembryonickidney293cellstoinvestigatetheroleofthecarboxylterminusofRKinrecognitionandphosphorylationofrhodopsin.RKC,likethewild-typeRK,wasdetectedinbothplasmamembranesandcytosolicfractions.TheCterminaltruncatedrhodopsinkinasewasunabletophosphorylatephoto-activatedrhodopsin,butpossesseskinaseactivitysimilartothewild-typeRKinphosphorylationofsmallpeptidesubstrate.ItsuggeststhatthetruncationdidnotdisturbthegrossstructuresofRKcatalyticdomain.OurresultsalsoshowthatRKCfailedtotranslocatetophoto-activatedrodoutsegments.Takentogether,ourstudydemonstratethecarboxylterminusofRKisrequiredforphosphorylationofphoto-activatedrhodopsinandstronglyindicatethatcarboxyl-terminusofRKmaybeinvolvedininteractionwithphoto-activatedrhodopsin.

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简介：AbstractBackground:Anterior thalamic nuclei (ATN) deep brain stimulation (DBS) is an effective method of controlling epilepsy, especially temporal lobe epilepsy. Mossy fiber sprouting (MFS) plays an indispensable role in the pathogenesis and progression of epilepsy, but the effect of ATN-DBS on MFS in the chronic stage of epilepsy and the potential underlying mechanisms are unknown. This study aimed to investigate the effect of ATN-DBS on MFS, as well as potential signaling pathways by a kainic acid (KA)-induced epileptic model.Methods:Twenty-four rhesus monkeys were randomly assigned to control, epilepsy (EP), EP-sham-DBS, and EP-DBS groups. KA was injected to establish the chronic epileptic model. The left ATN was implanted with a DBS lead and stimulated for 8 weeks. Enzyme-linked immunosorbent assay, Western blotting, and immunofluorescence staining were used to evaluate MFS and levels of potential molecular mediators in the hippocampus. One-way analysis of variance, followed by the Tukey post hoc correction, was used to analyze the statistical significance of differences among multiple groups.Results:ATN-DBS is found to significantly reduce seizure frequency in the chronic stage of epilepsy. The number of ectopic granule cells was reduced in monkeys that received ATN stimulation (P < 0.0001). Levels of 3′,5′-cyclic adenosine monophosphate (cAMP) and protein kinase A (PKA) in the hippocampus, together with Akt phosphorylation, were noticeably reduced in monkeys that received ATN stimulation (P = 0.0030 and P = 0.0001, respectively). ATN-DBS also significantly reduced MFS scores in the hippocampal dentate gyrus and CA3 sub-regions (all P < 0.0001).Conclusion:ATN-DBS is shown to down-regulate the cAMP/PKA signaling pathway and Akt phosphorylation and to reduce the number of ectopic granule cells, which may be associated with the reduced MFS in chronic epilepsy. The study provides further insights into the mechanism by which ATN-DBS reduces epileptic seizures.