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简介：Objective:Toevaluatetheosteogenicpotentialofbonemorphogeneticprotein(BMP)-2genetransfectedgoatbonemarrow-derivedmesenchymalstemcells(MSCs).Methods:Goatbonemarrow-derivedMSCsweretransfectedbyAdv-humanbonemorphogeneticprotein(hBMP)-2gene(Group1),Adv-betagaltransfectedMSCs(Group2)anduninfectedMSCs(Group3).Westernblotanalysis,alkalinephosphatasestaining,VonKossastainingandtransmissionelectronmicroscopywereadoptedtodeterminethephenotypeofMSCs.Thenthecellswereinjectedintothighmusclesofthenudemice.Radiographicalandhistologicalevaluationswereperformedatdifferentintervals.Results:OnlyAdv-hBMP-2transfectedMSCsproducedhBMP-2.Thesecellswerepositiveforalkalinephosphatasestainingatthe12thdayandwerepositiveforVonKossastainingatthe16thdayaftergenetransfer.Electronmicroscopicobservationshowedthatthereweremoreroughendoplasmicreticulum,mitochondriaandlysosomesinAdv-hBMP-2transfectedMSCscomparedtoMSCsofothertwogroups.Atthe3rdand6thweeksaftercellinjection,ectopicboneswereobservedinmusclesofnudemiceofGroup1.Onlyfibroustissueoralittlebonewasfoundinothertwogroups.Conclusions:BMP-2genetransfectedMSCscandifferentiateintoosteoblastsinvitroandinduceboneformationinvivo.

简介：OurpreliminarystudiesconfirmedthatanactiveprincipleregionofBuyangHuanwudecoction,comprisingalkaloid,polysaccharide,aglycon,glucosideandvolatileoil,caninducebonemarrowmesenchymalstemcelldifferentiationintoneurons.Mitogen-activatedproteinkinasesignalingwasidentifiedasoneofthekeypathwaysunderlyingthisdifferentiationprocess.Thepresentstudyshowsphosphorylatedextracellularsignal-regulatedproteinkinaseandphosphorylatedp38proteinexpressionwasincreasedafterdifferentiation.Cellularsignalingpathwayblockingagents,PD98059andSB203580,inhibitedextracellularsignal-regulatedproteinkinaseandp38inmitogen-activatedproteinkinasesignalingpathwaysrespectively.mRNAandproteinexpressionoftheneuronalmarker,neuronspecificenolase,andneuralstemcellmarker,nestin,weredecreasedinbonemarrowmesenchymalstemcellsaftertreatmentwiththeactiveprincipleregionofBuyangHuanwudecoction.Experimentalfindingsindicatethat,extracellularsignal-regulatedproteinkinaseandp38inmitogen-activatedproteinkinasesignalingpathwaysparticipateinbonemarrowmesenchymalstemcelldifferentiationintoneuron-likecells,inducedbytheactiveprincipleregionofBuyangHuanwudecoction.

简介：Wehaveconfirmedefficientanti-tumoractivitiesoftheperipherallymphocytestransducedwithap185HER2-specificchimericT-cellreceptorgenebothinmurineandinhumaninourpreviousstudies.TofurthertestthefeasibilityofchimericT-cellreceptorinabonemarrowtransplantationmodel,wefirst,madetwomurinetumorcelllines:MT901andMCA-205,toexpresshumanp185HER2byretroviralgenetransduction.MurinebonemarrowcellswereretrovirallytransducedtoexpressthechimericT-cellreceptorandgene-modifiedbonemarrowcellsweretransplantedintolethallyirradiatedmouse.Sixmonthsposttransplantation,p185HER2-positivetumorcells:MT-901/HER2orMCA-205/HER2wassubcutaneouslyorintravenouslyinjectedtomakemousemodelssimulatingprimarybreastcancerorpulmonarymetastasis.Theinvivoanti-tumoreffectsweremonitoredbythesizeofthesubcutaneoustumororcountingthetumornodulesinthelungsafterIndiainkstaining.ThesizeofthesubcutaneoustumorwassignificantlyinhibitedandthenumberofpulmonarynodulesweresignificantlydecreasedinmouserecipientstransplantedwithchimericT-cellreceptormodifiedbonemarrowcellscomparedwiththecontrolgroup.Ourresultssuggesttheefficientinvivoanti-tumoractivitiesofchimericT-cellreceptorgenemodifiedbonemarrowcells.

简介：BACKGROUND:Stereotacticinjection(striatumorlateralventricle)andvascularinjection(tailveinorcarotidartery)arenowoftenusedincellulartherapyforcerebralinfarction.Stereotacticinjectioncanaccuratelydelivercellstotheinfarctarea,butrequiresastereotacticdeviceandcausessecondarytrauma;vascularinjectioniseasyandbetterforhostneurologicaldeficitrecovery,butcancausethrombosis.OBJECTIVE:Tocomparethetherapeuticpotentialofadultbonemarrow-derivedmesenchymalstemcells(BMSCs)transplantationbyintraperitonealversusintravenousadministrationtocerebralischemicrats.DESIGN,TIMEANDSETTING:ArandomizedcontrolledanimalexperimentwasperformedattheCellRoomandPathologyLaboratory,BrainHospitalAffiliatedtoNanjingMedicalUniversityfromNovember2007toSeptember2008.MATERIALS:BMSCswerederivedfrom20healthySprague-Dawleyratsaged4-6weeks.METHODS:Forty-fiveadultmiddlecerebralarteryocclusion(MCAO)ratswererandomlydividedintocontrol,intravenousandintraperitonealinjectiongroups,with15ratsineachgroup.At21daysaftermodeling,ratsinthecontrolgroupreceived1mLof0.01mol/Lphosphatebufferedsalineviatailveininjectionandeachexperimentalratreceived4×106BMSCslabeledbybromodeoxyuridine(BrdU)viaintravenousorintraperitonealinjection.MAINOUTCOMEMEASURES:Angiogeninexpressionandsurvivaloftransplantedcellsweremeasuredbyimmunohistochemicalstainingofbraintissueininfarctionhemisphereat7,14or21daysafterBMSCtransplantation.Co-expressionofBrdU/microtubule-associatedprotein2orBrdU/glialfibrillaryacidicproteinwasobservedbydouble-labeledimmunofluorescenceofcerebralcortex.Evaluationofnervefunctionusingtheneurologicalinjuryseverityscoreandtheadhesion-removaltestwasperformedonthe1stand21stdaybeforeandafterMCAO,andat3,7,14or21daysafterBMSCstreatment.RESULTS:Angiogenin-positivenewvesselsweredistributedinthebilateralstriatum,hippocampusandcerebral

简介：摘要BackgroundDelayed encephalopathy (DE) is the most severe complication after acute carbon monoxide (CO) poisoning, which seriously affects the outcome of patients and leads to a high disability rate. Prior studies have shown that hyperbaric oxygen (HBO2) therapy is therapeutic for DE due to reducing immune-mediated neuropathology and thus improving cognitive performance.MethodsIn our present perspective study, five DE patients were treated regularly with HBO2 therapy. The mini-mental state examination (MMSE) and Barthel index (BI) were intermittently collected during their hospitalization for mental and physical status evaluation, the peripheral bloods were serially sampled to determine the concentration changes of circulating stem cells, as well as corresponding BDNF and neural markers.ResultsMMSE and BI showed series of improvements after multiple HBO2 therapies. The CD34+/CD90+ and CD34+/CD133+ dual positive cells, which were categorized as circulating stem cells, were observed an overall up-regulation since the beginning of the DE onset upon the application of HBO2 therapy. Characteristic neurotrophin BDNF, neural markers such as nestin and synaptophysin (SYP) were also up-regulated after exposure of HBO2. Conclusion The application of HBO2 therapy is of significance in improving the cognition of DE patients, along with mobilized circulating stem cells.ConclusionWe primarily infer that the CD34+/CD90+ and CD34+/CD133+ cells were mobilized by HBO2 exposure and have played a positive role in cognition improvement on DE patients by up-regulation of BDNF, nestin and SYP. The altering amount of circulating stem cells mobilized in peripheral blood could be a potential marker on predicting the outcome of DE.

简介：AbstractBackground:Investigations of the pathogenic mechanisms in motor neurons (MNs) derived from amyotrophic lateral sclerosis (ALS) disease-specific induced pluripotent stem (iPS) cell lines could improve understanding of the issues affecting MNs. Therefore, in this study we explored mutant superoxide dismutase 1 (SOD1) protein expression in MNs derived from the iPS cell lines of ALS patients carrying different SOD1 mutations.Methods:We generated induced pluripotent stem cell (iPSC) lines from two familial ALS (FALS) patients with SOD1-V14M and SOD1-C111Y mutations, and then differentiated them into MNs. We investigated levels of the SOD1 protein in iPSCs and MNs, the intracellular Ca2+ levels in MNs, and the lactate dehydrogenase (LDH) activity in the process of differentiation into the MNs derived from the controls and ALS patients’ iPSCs.Results:The iPSCs from the two FALS patients were capable of differentiation into MNs carrying different SOD1 mutations and differentially expressed MN markers. We detected high SOD1 protein expression and high intracellular calcium levels in both the MN and iPSCs that were derived from the two SOD1 mutant patients. However, at no time did we observe stronger LDH activity in the patient lines compared with the control lines.Conclusions:MNs derived from patient-specific iPSC lines can recapitulate key aspects of ALS pathogenesis, providing a cell-based disease model to further elucidate disease pathogenesis and explore gene repair coupled with cell-replacement therapy. Incremental mutant expressions of SOD1 in MNs may have disrupted MN function, either causing or contributing to the intracellular calcium disturbances, which could lead to the occurrence and development of the disease.

简介：AbstractBackground:The role of allogeneic hematopoietic stem cell transplantation (allo-HSCT) in children with high-risk (HR) T-cell acute lymphoblastic leukemia (T-ALL) in first complete remission (CR1) is still under evaluation. Moreover, relapse is the main factor affecting survival. This study aimed to explore the effect of allo-HSCT (especially haploidentical HSCT [haplo-HSCT]) on improving survival and reducing relapse for HR childhood T-ALL in CR1 and the prognostic factors of childhood T-ALL in order to identify who could benefit from HSCT.Methods:A total of 74 newly diagnosed pediatric T-ALL patients between January 1, 2012 and June 30, 2018 were enrolled in this retrospective study. Patients were stratified into the low-risk chemotherapy cohort (n = 16), HR chemotherapy cohort (n = 31), and HR transplant cohort (n = 27). Characteristics, survival outcomes, and prognostic factors of all patients were then analyzed.Results:Patient prognosis in the HR chemotherapy cohort was significantly worse than that in the low-risk chemotherapy cohort (5-year overall survival [OS]: 58.5% vs. 100%, P = 0.003; 5-year event-free survival [EFS]: 54.1% vs. 83.4%, P = 0.010; 5-year cumulative incidence of relapse [CIR]: 45.2% vs. 6.3%, P = 0.011). In HR patients, allo-HSCT improved the 5-year EFS and CIR compared to that of chemotherapy (5-year EFS: 80.1% vs. 54.1%, P = 0.041; 5-year CIR: 11.6% vs. 45.2%, P = 0.006). The 5-year OS was higher in the HR transplant cohort than that in the HR chemotherapy cohort (81.0% vs. 58.5%, P = 0.084). Minimal residual disease re-emergence was an independent risk factor for 5-year OS, EFS, and CIR; age ≥10 years was an independent risk factor for OS and EFS; and high white blood cell count was an independent risk factor for EFS and CIR.Conclusion:Allo-HSCT, especially haplo-HSCT, could effectively reduce relapse of children with HR T-ALL in CR1.

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简介：在发炎和白血球过多症之间的潜在的交叉连接有一段时间被讨论了，但是支持这教义的试验性的证据是少见的。特别地，由煽动性的调停人理解为proto-oncogenic生长因素表情的潜在的upregulation负责的机制是重要的。这里，我们调查了高度煽动性的cytokineinterleukin-1贝它的能力(IL-1β；)到导致干细胞因素(SCF)的生产，它是在造血的myeloid细胞的恶意的转变之上控制尖锐myeloid白血球过多症的前进的一个主要造血的生长因素。我们发现了那人的IL-1β；在MCF-7人的上皮的乳癌房间和那导致了SCF的表示/分泌物这个过程取决于组织缺氧可诱导的因素1(HIF-1)抄写建筑群。我们也表明了phosphatidylinositol-3kinase(PI-3K)的一个关键角色rapamycin(mTOR)的/mammalian目标在IL-1β的小径；导致的HIF-1α；在MCF-7房间的累积。重要地，mTOR也被发现在IL-1β起一个作用；导致的SCF生产。而且，为IL-1β的积极关联的一个趋势；并且在健康人的施主的血浆的SCF层次被观察。总的来说，我们的结果表明那IL-1β；，它通常衔接天生、适应的免疫，通过PI-3K/mTOR小径和HIF-1抄写建筑群导致主要haematopoietic/proleukaemic生长因素SCF的生产。这些调查结果强烈支持在发炎和尖锐myeloid白血球过多症之间的串音。

简介：BACKGROUND:Alpha-actinin(α-actinin)playsakeyroleinneuronalgrowthconemigrationduringdirectionaldifferentiationfromneuralstemcells(NSCs)toneurons.OBJECTIVE:Todetectinsitumicrodistributionandquantitativeexpressionofα-actininduringdirectionaldifferentiationofNSCstoneuronsinthetemporallobecerebralcortexofneonatalrats.DESIGN,TIMEANDSETTING:BetweenJanuary2006andDecember2008,cultureanddirectionaldifferentiationofNSCswereperformedatDepartmentofHistologyandEmbryology,PreclinicalMedicalCollege,ChinaMedicalUniversity.ImmuneelectronmicroscopywasperformedatDepartmentofHistologyandEmbryologyandDepartmentofElectronMicrology,PreclinicalMedicalCollege,ChinaMedicalUniversity.SpectrumanalysiswasperformedatLaboratoryofElectronMicroscopy,MentalResearchInstitute,ChineseAcademyofSciences.MATERIALS:Basicfibroblastgrowthfactor,epidermalgrowthfactor,brain-derivednervegrowthfactor,type-1insulinlikegrowthfactor,andα-actininantibodywereprovidedbyGibcoBRL,USA;rabbit-anti-ratnestinmonoclonalantibody,rabbit-anti-ratneuronspecificenolasepolyclonalantibody,andEDAX-9100energydispersiveX-rayanalysiswereprovidedbyPHILIPSCompany,Netherlands.METHODS:NSCs,followingprimaryandpassageculture,weredifferentiatedwithserumculturemedium(DMEM/F_(12)+10%fetalbovineserum+2ng/mLbrain-derivednervegrowthfactor+2ng/mLtype-1insulinlikegrowthfactor).MAINOUTCOMEMEASURES:Expressionofα-actinininneuron-likecellswasquantitativelyandqualitativelydetectedwithimmunocytochemistryusingenergydispersiveX-rayanalysis.RESULTS:Immunocytochemistry,combinedwithelectronmicroscopy,indicatedthatpositiveα-actininexpressionwaslikeaspheroidparticlewithhighelectrondensity.Inaddition,theexpressionwasgraduallyconcentratedfromthenuclearedgetothecytoplasmandexpandedintodevelopingneurites,duringdifferentiationofneuralstemcellstoneurons.Conversely,energydispersive

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.

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.