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简介：Objective:Toobservetheeffectsofthermalstressonproliferationofhumanvascularendothelialcells(VECs)andexploreitssignificance.Methods:ChangesofVECsproliferationwereinvestigatedwith3H-TdRincorporationmethodafterECV304wastreatedat43℃for2hours,whileexpressionsofintercellularadhesionmolecule-1(ICAM-1),inhibitorofdifferentiation-1(ID1),andP16andP21proteinsweredeterminedbyWesternBlotting.Results:TheeffectofinhibitionofVECsgrowthafterthermalstresswasdetectedby3H-TdRincorporationexperiment.WesternblottingshowedICAM-1,amarkerofactivatedendothelialcells,wasincreasedmarkedlyafterthermalstress.ExpressionofID1proteindeclinedgraduallywithincreasingexpressionsofitsdownstreamgenes,P16andP21followingthethermalstress.Conclusions:ThermalstresscouldstronglyactivateVECsandinhibitproliferationofVECsthroughID1,thusdownregulatingcyclin-dependentkinaseinhibitors,P16andP21,whichmightbeanessentialpathwayforrecoveryofVECsafterthermalstress.

简介：客观CXCR4是为癌症基因治疗的一个潜在的目标。在这研究，指向CXCR4基因的RNA干扰制动火箭病毒向量被构造，并且它对由压抑CXCR4基因表示的前列腺癌症房间的侵略的影响被分析。CXCR4特定的siRNA基因被PCR克隆并且插入了进Pgensil-1原生质标志反对染色U6倡导者和EGFP的方法，然后recombinant碎裂是进PLXSN的亚cloned并且由限制酶测试了并且定序。从transfectedPA317房间获得的病毒是进前列腺癌症房间的transfected。CXCR4mRNA的表示被RT-PCR检测。前列腺癌症房间在试管内的侵略的能力被Transwell实验估计。与recombinantPLXSNtransfection，在前列腺癌症房间PC-3m和LNCaP的CXCR4mRNA的表示以率被减少，这被显示出的结果(84.26±10.20)%并且(88.17±11.23)%分别地。Transwell实验的结果也证明房间在下面的数字微膜分别地，在控制组(P<0.05)与46.9±5.3和66.7±6.0相比在PC-3m和LNCaP的对待的组是14.7±3.1和18.9±4.2。结论PLXSN/EGFP-U6-siCXCR4能显著地压抑表示fCXCR4，前列腺癌症房间在试管内的侵略海角被也禁止。这recombinant碎片将在前列腺癌症的治疗是有用的。

简介：Antifungaldrugresistanceisasignificantclinicalproblem,andantifungalagentsthatcanevaderesistanceareurgentlyneeded.Ininfectiveniches,resistantorganismsoftenco-existedwithsensitiveones,orasubpopulationofantibiotic-susceptibleorganismsmayevolveintoresistantonesduringantibiotictreatmentandeventuallydominatethewholepopulation.Inthisstudy,weestablishedaco-cultureassayinwhichanazole-resistantCandidaalbicansstrainwasmixedwithasusceptiblestrainlabeledwithgreenfluorescentproteintomimicinvivoconditionsandscreenforantifungaldrugs.Fluconazolewasusedasapositivecontroltoverifythevalidityofthisco-cultureassay.FivenaturalmoleculesexhibitedantifungalactivityagainstbothsusceptibleandresistantC.albicans.Twoofthesecompounds,retigericacidB(RAB)andriccardinD(RD),preferentiallyinhibitedC.albicansstrainsinwhichtheeffluxpumpMDR1wasactivated.Thisselectivitywasattributedtogreaterintracellularaccumulationofthedrugsintheresistantstrains.Changesinsterolandlipidcompositionswereobservedintheresistantstrainscomparedtothesusceptiblestrain,andmightincreasecellpermeabilitytoRABandRD.Inaddition,RABandRDinterferedwiththesterolpathway,furtheraggregatingthedecreaseinergosterolinthesterolsynthesispathwayintheMDR1-activatedstrains.Ourfindingshereprovideanalternativeforcombatingresistantpathogenicfungi.

简介：AIMMacrophagemigrationinhibitoryfactor(MIF)isapro-inflammatorycytokineinvolvedinthepathogenesisofavarietyofautoimmuneandinflammatorydiseases,includingarthritis,glomerulonephritis,Gram-positiveandGram-negativesepsis,andatherogenesis.RecentstudiesshowedthatCD74(antigen-associatedinvariantchainⅡ)isahigh-affinitymembrane-bindingproteinforMIF.ThepurposeofthepresentstudywastoexpresstherecombinanthumanCD74inE.coliandinhibittheactivityofMIFbyusingrecombinantCD74invitro.

简介：Recently,ithasbeenreportedthattheluteolyticac-tionwasthemainmechanismoftheterminationofearlypregnancybycontraceptivest.Inaddition,someestro-genreceptorligandssuchasdroloxifenehavebeenshowntoinhibitthegrowthofculturedratlutealcells.Ithasbeenknownthat2-phenylindolederivativescouldcompetewithestradiolfortheestrogenreceptor.Theseresultspromptedustodesignandsynthesizethefollow-ingsixnew3-oxamoyl-2-phenylindolecompounds1a-fandtesttheireffectsonthegrowthoflutealcells,aimingatseekingnewleadcompoundspossessingantifertilityactivities.

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简介：AbstractObjective:Glioma is the most common and aggressive primary brain tumor. Here, we aimed to establish a nano-drug carrier system to traverse the blood-brain barrier for the treatment and inhibition of glioma migration.Methods:The synthesis of bovine serum albumin protected-silver nanoclusters (BSA-AgNCs) was performed using chemical reduction. The drug paclitaxel (PTX) can be loaded into BSA-AgNCs through electrostatic and hydrophobic interactions to formulate spherical BSA-AgNC-PTX nanoparticles (BSA-AgNC-PTX NPs). A glioma mouse model was established by injecting U251-GFP-Luc cells into the mouse striatum, and all surgical procedures were approved by the Animal Ethics Committee of Nanchang University (SYXK2019-0003) on December 29, 2019.Results:The BSA-AgNC-PTX NPs were able to efficiently pass through the blood-brain barrier, both in vitro and in vivo, to deliver the drug to the tumor site. The in vivo assessment of BSA-AgNC-PTX NPs in glioblastoma multiforme-bearing mice revealed the significant inhibition of tumor growth and migration, prolonging the survival of the mice.Conclusion:These results indicated that BSA-AgNCs might represent an ideal nanocarrier for the treatment of glioma and has the potential to be used in the treatment of a variety of central nervous system diseases.

简介：这studay是构造lentiviral向量harbouring指向编码人的高活动性的组nucleosomal绑定蛋白质的基因的一个RNA干扰(RNAi)序列1(NSBP1)；为了在导致G2/M学习它的角色，在前列腺癌症(PCa)分阶段执行拘捕和apoptosisDU145房间；并且估计效果它的在在vitro并且在vivo的房间增长上击倒。RNAi被使用由生产NSBP1的lentiviralplasmids在PCa房间线DU145将NSBP1表示击倒小发卡RNA。在在DU145细胞击倒的NSBP1以后，细胞的生长率被MTT，和G2/M细胞周期拘捕分析，apoptosis用FACScalibur流动cytometer被估计。瘤生长在裸体老鼠被估计。NSBP1，cyclinB1和Bcl-2的mRNA和蛋白质表示层次被reverse-transcriptase聚合酶链反应并且西方的弄污在vitro并且在vivo分析。NSBP1击倒在96h，在裸体老鼠的减少的瘤生长，和G2/M房间周期拘捕(8.78%)和apoptosis(2.19褶层)的正式就职与PscNClentivirus控制房间相比处于房间的生长率导致了22.6%减少。与房间周期拘捕和apoptosis一致，cyclinB1和Bcl-2的mRNA和蛋白质表示层次被减少。在结论，NSBP1原因击倒统计上重要的抑制在里面vitro并且在PCa细胞线DU145的vivo生长。生长抑制是部分至少由于NSBP1击倒导致G2/M房间骑车拘捕和apoptosis。现在的数据提供NSBP1击倒导致G2/M阶段拘捕和apoptosis可以由NSBP1源于cyclinB1和Bcl-2的否定规定的证据，与结果这些蛋白质的减少的表示。

简介：AbstractObjective:This study was performed to investigate the relationship between the human melanogenesis and antioxidant systems and to further confirm the synergistic effect of oxyresveratrol (OXYR) and resveratrol (RES) in human epidermal melanocyte cell line.Methods:The human epidermal melanocyte line PIG1 cells were divided into the UV groups and control group, treated with different doses of UVB and without UVB, respectively. MTT assay and flow cytometry were used to detect cell viability and apoptosis. The expression of Nrf2/HO-1 and melanogenesis-associated proteins/genes was measured by Western blotting and real-time qPCR (RT-qPCR). pCMV6-XL5-Nrf2 was used to upregulate the expression of Nrf2. Subsequently, the proteins/genes levels of Nrf2/HO-1 and tyrosinase (TYR), melanin/eumelanin content, and reactive oxygen species (ROS) were analyzed. Isobologram analysis and cell experiment were used to analyze whether OXYR and RES inhibit TYR synergistically. Western blotting, RT-qPCR, and NaOH splitting method were used to determine the Nrf2/HO-1 and melanogenesis-associated proteins/genes expression and melanin content to evaluate the efficacy of OXYR and RES.Results:The activated Nrf2 and HO-1 eliminated ROS produced by UVB irradiation. The melanogenesis-associated proteins/genes of melanocyte-inducing transcription factor (MITF, P < 0.01 on protein expression), TYR (both P < 0.01), TYR-related protein (TRP)-1 (both P < 0.05), and TRP2 (P < 0.05 on mRNA expression) were activated in PIG1 cells by UVB irradiation. Simultaneously, the upregulation of Nrf2 significantly reduced melanogenesis formation (P < 0.001) and TYR level (P < 0.01 on protein expression). Moreover, OXYR and RES synergistically inhibited TYR activity (P < 0.001) and reduced melanin content (P < 0.001).Conclusions:A microbalance exists between Nrf2/HO-1 signaling and melanogenesis production in the UVB-induced responses of melanocytes. Simultaneously, OXYR enhances the ability of RES to inhibit melanin production.

简介：树枝状的房间(DC)有力量通过有免疫力的回答的调整调整autoimmunity的结果。抗原的正式就职特定的忍耐为allograft拒绝的预防和处理是批评的。在现在的学习，我们transfectedIL-10基因进DC并且在老鼠在小岛allograft上在有免疫力的忍耐的vitro和正式就职在淋巴细胞活动的抑制上调查了他们的效果。一个IDDMC57BL/6老鼠模特儿被streptozotocin劝诱。从BALB/c老鼠孤立的小岛房间被移植进老鼠由IL-10的注射跟随了的模型的肾capules修改DC(mDCs)。结果证明mDCs能显著地禁止T淋巴细胞增长由同种抗免疫球蛋白房间调停了并且导致它的apoptosis，而，未修改的DC(umDCs)能显著地支持鼠科的淋巴细胞增长。mDCs的注射能延长同种抗免疫球蛋白小岛的幸存移植IDDM老鼠。在mDCs的平均血浆葡萄糖(PG)水平对待在3天以内还给正常并且持续了大约2个星期的老鼠。在控制老鼠的拒绝反应在移植以后发生了5天。当IL-4在mDCs是更高的时，IFN-的水平更低在umDCs比那对待老鼠对待老鼠，它显示Th1/Th2偏差发生了。我们的研究建议那IL-10基因修改了DC能导致有免疫力的忍耐到小岛接枝并且由在同种抗免疫球蛋白老鼠禁止T房间增长延长接受者的幸存。

简介：一个entomopathogenic细菌，Xenorhabdusnematophila，被知道了由禁止联系免疫者的phospholipaseA2(PLA2)导致目标昆虫的重要免疫力的抑制，它随后关掉在在昆虫的有免疫力的调停是批评的eicosanoids的生合成。一些代谢物从细菌的文化肉汤发源被识别了并且包括benzylideneacetone，脯氨酸酷氨酸和acetylatedphenylalanine-glycine-valine，它被知道禁止从血球和胖身体提取的PLA2的酶活动。这研究在甜菜粟蚕的消化PLA2上测试了他们的效果，Spodopteraexigua。年轻幼虫喂了三代谢物的不同集中在100g/mL下面甚至在剂量在幼虫的开发导致了重要不利效果。特别地，他们在摄取的食物的消化效率导致了重要减小。所有三代谢物显著地禁止了在一个低micromolar范围从第五个中间形态幼虫的midgut腔提取的消化PLA2的催化活动。这些结果建议S的消化PLA2上的三细菌的代谢物的禁止的活动。exiguamidgut可以解释一些他们的口头的有毒的效果。

简介：Objective:Toinvestigatetheinvitrocleavageabilityandeffectsonapoptosisandcellgrowthofthebcr-ablfusiongenespecificmulti-unitribozymes.Methods:Threefusionpointspecificribozymesweredesignedandthemulti-unitribozymes'invitrotranscriptionvectorandretroviralvectorwereconstructed.Theinvitrocleavageabilitywastested.TheretroviralvectorwastransfectedintoK562cellandtheeffectsonproliferation,apoptosis,cellcycleandcellstructurewereobserved.Results:Multi-unitribozymeshadinvitrocleavageefficiencyof70.8%,whichwasmoreefficientthansingle-unitanddouble-unitribozymes.TransfectionoftheretroviralvectoroftheribozymeintoK562cells,inducedinhibitionofcellgrowthandapoptosis.TheincorporationrateofDNAinribozymestransfectedK562cellswasgreatlydecreasedalongwithtimepassed,withaninhibitionrateofmorethan50%after96hoftransfection.UnderFCM,18.4%ofthecellsunderwentapoptosis72haftertransfectionandmorecellswereblockedinGphase,withtheratioinSphasegreatlydecreased(41.9%).Underelectronmicroscope,compactionofnuclearchromatinandapoptosisbodieswereobserved.Conclusion:Multi-unitribozymesspecifictobcr-ablfusiongenecanbeusedtotreatCMLandtopurgebonemarrowforself-grafting.

简介：cAMPmediatedsignalingmayplayasuppressiveroleinimmuneresponse.WepreviouslyfoundthatthecAMP-elevators(CTxand8-Br-cAMP)inhibitedIL-12,IL-la,IL-6geneexpression,butincreasedthetranscriptionallevelsofIL-10andIL-1RainLPS-treatedmurineperitonealmacrophages.ThepresentstudyexaminedapossiblemolecularmechanisminvolvedincAMPelevators-inducedinhibitionofIL-12p40expressioninresponsetoLPS.OurdatademonstratedthatcAMPelevatorsdownregulatedIL-12p40mRNAexpressionandIL-12pT0productioninmurineperitonealmacrophages.SubsequentstudiesrevealedthatcAMP-elevatorsblockedphosphorylationofp38MAPK,butdidnotaffecttheactivityofNF-κBbindingtoIL-12promoter(-136/-112).ThisisthefirstreportthatcAMPelevatorsinhibitLPS-inducedIL-12productionbyamechanismthatisassociated,atleastinpart,withp38-dependentinhibitionbycAMPsignalingpathways.

简介：AbstractBackground:Emerging evidence indicates that the sineoculis homeobox homolog 1-eyes absent homolog 1 (SIX1-EYA1) transcriptional complex significantly contributes to the pathogenesis of multiple cancers by mediating the expression of genes involved in different biological processes, such as cell-cycle progression and metastasis. However, the roles of the SIX1-EYA1 transcriptional complex and its targets in colorectal cancer (CRC) are still being investigated. This study aimed to investigate the roles of SIX1-EYA1 in the pathogenesis of CRC, to screen inhibitors disrupting the SIX1-EYA1 interaction and to evaluate the efficiency of small molecules in the inhibition of CRC cell growth.Methods:Real-time quantitative polymerase chain reaction and western blotting were performed to examine gene and protein levels in CRC cells and clinical tissues (collected from CRC patients who underwent surgery in the Department of Integrated Traditional and Western Medicine, West China Hospital of Sichuan University, between 2016 and 2018, n = 24). In vivo immunoprecipitation and in vitro pulldown assays were carried out to determine SIX1-EYA1 interaction. Cell proliferation, cell survival, and cell invasion were determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, clonogenic assay, and Boyden chamber assay, respectively. The Amplified Luminescent Proximity Homogeneous Assay Screen (AlphaScreen) method was used to obtain small molecules that specifically disrupted SIX1-EYA1 interaction. CRC cells harboring different levels of SIX1/EYA1 were injected into nude mice to establish tumor xenografts, and small molecules were also injected into mice to evaluate their efficiency to inhibit tumor growth.Results:Both SIX1 and EYA1 were overexpressed in CRC cancerous tissues (for SIX1, 7.47 ± 3.54 vs.1.88 ± 0.35, t = 4.92, P = 0.008; for EYA1, 7.61 ± 2.03 vs. 2.22 ± 0.45, t = 6.73, P = 0.005). The SIX1/EYA1 complex could mediate the expression of two important genes including cyclin A1 (CCNA1) and transforming growth factor beta 1 (TGFB1) by binding to the myocyte enhancer factor 3 consensus. Knockdown of both SIX1 and EYA1 could decrease cell proliferation, cell invasion, tumor growth, and in vivo tumor growth (all P < 0.01). Two small molecules, NSC0191 and NSC0933, were obtained using AlphaScreen and they could significantly inhibit the SIX1-EYA1 interaction with a half-maximal inhibitory concentration (IC50) of 12.60 ± 1.15 μmol/L and 83.43 ± 7.24 μmol/L, respectively. Administration of these two compounds could significantly repress the expression of CCNA1 and TGFB1 and inhibit the growth of CRC cells in vitro and in vivo.Conclusions:Overexpression of the SIX1/EYA1 complex transactivated the expression of CCNA1 and TGFB1, causing the pathogenesis of CRC. Pharmacological inhibition of the SIX1-EYA1 interaction with NSC0191 and NSC0933 significantly inhibited CRC cell growth by affecting cell-cycle progression and metastasis.

简介：AbstractBackground:The use of microRNAs in the therapy of kidney disease is hampered by the difficulties in their effective delivery. Microvesicles (MVs) are known as natural carriers of small RNAs. Our prior research has demonstrated that MVs isolated from mesenchymal stem cells (MSCs) are capable of attenuating kidney injuries induced by unilateral ureteral obstruction and 5/6 sub-total nephrectomy in mice. The present study aimed to evaluate the effects of miR-34a-5p (miR-34a)-modified MSC-MVs on transforming growth factor (TGF)-β1-induced fibrosis and apoptosis in vitro.Methods:Bone marrow MSCs were modified by lentiviruses over-expressing miR-34a, from which MVs were collected for the treatment of human Kidney-2 (HK-2) renal tubular cells exposed to TGF-β1 (6 ng/mL). The survival of HK-2 cells was determined using 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) and Annexin V-Light 650/propidium iodide (PI) assays. The expression levels of epithelial markers (tight junction protein 1 [TJP1] and E-cadherin) and mesenchymal markers (smooth muscle actin alpha (α-SMA) and fibronectin) in HK-2 cells were measured using Western blot analysis and an immunofluorescence assay. In addition, changes in Notch-1/Jagged-1 signaling were analyzed using Western blotting. Data were analyzed using a Student’s t test or one-way analysis of variance.Results:MiR-34a expression increased three-fold in MVs generated by miR-34a-modified MSCs compared with that expressed in control MVs (P < 0.01, t= 16.55). In HK-2 cells, TJP1 and E-cadherin levels decreased to 31% and 37% after treatment with TGF-β1, respectively, and were restored to 62% and 70% by miR-34a-enriched MSC-MVs, respectively. The expression of α-SMA and fibronectin increased by 3.9- and 5.0-fold following TGF-β1 treatment, and decreased to 2.0- and 1.7-fold after treatment of HK-2 cells with miR-34a-enriched MSC-MVs. The effects of miR-34a-enriched MSC-MVs on epithelial-mesenchymal transition (EMT) markers were stronger than control MSC-MVs. The effects of miR-34a-enriched MSC-MVs on these EMT markers were stronger than control MSC-MVs. Notch-1 receptor and Jagged-1 ligand, two major molecules of Notch signaling pathway, are predicted targets of miR-34a. It was further observed that elevation of Notch-1 and Jagged-1 induced by TGF-β1 was inhibited by miR-34a-enriched MSC-MVs. In addition, TGF-β1 exposure also induced apoptosis in HK-2 cells. Although miR-34a-mofidied MSC-MVs were able to inhibit TGF-β1-triggered apoptosis in HK-2 cells, the effects were less significant than control MSC-MVs (control:TGF-β1 :miR-nc-MV:miR-34a-MV = 1.3:0.6:1.1:0.9 for MTT assay, 1.8%:23.3%:9.4%:17.4% for apoptosis assay). This phenomenon may be the result of the pro-apoptotic effects of miR-34a.Conclusions:The present study demonstrated that miR-34a-over-expressing MSC-MVs inhibit EMT induced by pro-fibrotic TGF-β1 in renal tubular epithelial cells, possibly through inhibition of the Jagged-1/Notch-1 pathway. Genetic modification of MSC-MVs with an anti-fibrotic molecule may represent a novel strategy for the treatment of renal injuries.