简介：BACKGROUND:Highincidenceofstrokeatinterchangeperiodofautumnandwinterwasdemonstratedbyepidemiologicalsurvey,andthespecificcausesshouldbefurtherinvestigated.OBJECTIVE:Toinvestigatetheinfluenceofartificialcoldexposureontheincidenceofstrokeinrenovascularhypertensiverats(RHR),andanalyzetheassociationwithbloodpressureandcold-inducibleRNAbindingprotein(CIRP)mRNAexpressioninbraintissue.DESIGN:Acompletelyrandomizedgroupingdesign,arandomizedcontrolanimaltrial.SETTINGS:LabofNeurology,theFirstAffiliatedHospitalofSunYat-senUniversity;DepartmentofChemistry,OpenlaboratoryofChemicalBiology,InstituteofMolecularTechnologyforDrugDiscoveryandSynthesis,UniversityofHongKong.MATERIALS:MaleSDrats(n=460),weighing80-100gwereobtainedfromGuangdongProvinceHealthAnimalUnit.AmodifiedRXZ-300Aintelligentartificialclimatecabinet(NingboJiangnanInstrumentCo.,Ltd.,China).METHODS:TheexperimentwereprocessedintheLabofNeurology,theFirstAffiliatedHospitalofSunYat-senUniversityandtheOpenLaboratoryofChemicalBiology,InstituteofMolecularTechnologyforDrugDiscoveryandSynthesis,UniversityofHongKongfromOctober2004toNovember2005.Rats(n=400)wereoperatedtoestablish2-kidney2-clipRHRmodelasdescribedpreviously.Thesham-operatedrats(n=60)servedasnormotensivecontrols.Eightweekslater,300ofRHRwererandomlyselectedaccordingtotheirsystolicbloodpressure(SBP)anddividedinto3sub-groups(n=100pergroup):mildhypertensivegroup(SBPof160-200mmHg),moderatehypertensivegroup(SBPof200-220mmHg)andseverehypertensivegroup(SBP>220mmHg).Eachgroupwasfurtherdividedintotwogroups(n=50)underACEandnon-ACE.Normalsham-operatedSDrats(n=60),SBP<140mmHg,wererandomlydividedintotwogroups:Sham-operatedcontrolgroup(n=30)underACEandnon-ACE.ToestablishtheACEandnon-ACEtreatment,ratswerehousedindividuallyinartifici

简介：InordertoinvestigatewhetherlipoxinA4(LXA4)hasanantagonisticeffectonlipopolysaccharide(LPS)-inducedsynthesisofinterleukin(IL)-1β,IL-6andIL-8inratpulmonarymicrovascularendothelialcells(PMVEC),andtoexplorethemolecularmechanismsofsignalpathwayinLXA4actions,culturedPMVECweretreatedwithLPS,withorwithoutpreincubationwithLXA4.ProteinsofIL-1β,IL-6andIL-8insupernatantwereanalyzedbyenzyme-linkedimmunosorbentassay(ELISA).ExpressionsofmRNAofIL-1β,IL-6andIL-8weredeterminedbyRT-PCR.Expressionsofphosphorylationofphosphoinositide3-kinase(PI3-K)andmyeloiddifferentiationfactor88(MyD88)wereanalyzedbyWesternblot.ActivitiesofDNA-bindingofnuclearfactor-kappaB(NF-κB)andactivatorprotein-1(AP-1)weremeasuredbyelectrophoreticmobilityshiftassay(EMSA).TheresultsshowedthatLPSinducedproductionofIL-1β,IL-6andIL-8inratPMVECviaMyD88/PI3-K/NF-κBandAP-1pathway-dependentsignaltransduction.LPS-stimulatedexpressionofPI3-K,activitiesofNFκBandAP-1,secretionofproteinandexpressionofmRNAofIL-1β,IL-6andIL-8butnotMyD88expressioninPMVECwereinhibitedbyLXA4inadose-dependentmanner.Inconclusion,LXA4inhibitssynthesisofIL-1β,IL-6andIL-8bydown-regulationofPI3-K/NF-κBandAP-1signalpathwayinPMVEC.

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简介：调查在在hypoxic下面的gliomaangiogenesis的受体(u-PA/u-PAR)系统调节的尿激类型plasminogenactivator/urokinase-typeplasminogen的角色的目的，我们在导致的组织缺氧上学习了调节glioma的媒介的效果在人的象endothelial一样ECV304房间增长，apoptosis，在vitro的绳索形成和u-PA/u-PAR表示的变化。方法MTT试金被用来在房间增长检验变化。房间apoptosis被染色的Hoechst33258分析。Matrigel像绳索的形成试金被用来在vitro评估ECV304房间的angiogenesis能力。u-PA/u-PARmRNA的表情被量的即时RT-PCR检测。结果组织缺氧禁止了ECV304房间增长并且导致了房间apoptosis。当不normoxic调节了部分堵住的U251glioma房间的媒介(N厘米)时，Hypoxic调节了媒介(H厘米)ECV304房间增长和apoptosis上的组织缺氧的效果。U251glioma房间的H厘米也支持了在matrigel上播种的ECV304房间的绳索形成。当u-PA或u同等monoclonal抗体被增加进ECV304房间culturing媒介时，绳索形成能力部分被禁止。U251glioma房间的H厘米在ECV304房间导致了uPA和uPAR表示。这些建议那个u-PA/u-PAR系统的结论被hypoxicmicroenviroment涉及gliomaangiogenesistrigged。

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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.

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简介：AbstractBackground:Baicalein has been shown to have anti-inflammatory and anti-tumor activities. However, the mechanisms underlying its anti-inflammatory effect on colitis remain unclear.Methods:A dextran sodium sulfate (DSS)-induced model of acute colitis was established in BALB/c mice (6-8 weeks old, weighing 18-22 g). Six groups of mice received: (1) water for 10 days (control), n = 6; (2) DSS 4% solution in the drinking water for 7 days, followed by normal water for 3 days, n = 7; (3), (4), and (5) as for group 2 plus baicalein (10, 20, 40 mg/kg) administered once daily starting on day 1, n = 6; and (6) as for (2) plus 5-aminosalicylic acid (50 mg/kg) administered once daily starting on day 1, n = 6. Body weights, stool consistency, and hematochezia were recorded, and the severity of colitis was evaluated using a disease activity index. On day 11, the mice were euthanized, and organs and blood were collected for analysis. Serum inflammatory factors were detected by enzyme-linked immunosorbent assay; CD11b-positive cells were analyzed by immunofluorescence microscopy; expression of retinoic-acid-receptor-related orphan nuclear receptor gamma, sphingosine kinase 1 (SPHK1), and phosphorylated signal transducer and activator of transcription 3 (p-STAT3) was detected by immunohistochemistry; and expression of nucleotide-binding oligomerization domain 2 (NOD2), SPHK1, sphingosine 1-phosphate receptor 1 (S1PR1), total STAT3, and p-STAT3 were detected by western blotting analysis. Inter-group differences were compared using Student’s t test.Results:Baicalein treatment dose-dependently reduced DSS-induced weight loss (P < 0.01 or P < 0.05), splenomegaly (P < 0.01), and colonic damage, as reflected by amelioration of diarrhea, rectal bleeding, and colonic ulceration, congestion, edema (shown as colon length, P < 0.05 or P < 0.01), and inflammatory cell infiltration. Baicalein also significantly decreased the levels of inflammatory mediators in the serum (P < 0.01) and colon, and significantly inhibited expression of NOD2 SPHK1, S1PR1, and p-STAT3 in the colon (P < 0.05).Conclusions:Baicalein treatment ameliorated colitis in mice by inhibiting S1P-STAT3 signaling, suggesting that this flavonoid might be beneficial in the treatment of colitis.

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简介：Objectives:Apoptosisisrecognizedasanimportantmechanismincontrast-inducednephropathy(CIN).Cordycepssinensis(CS),atime-honoredtonicfoodandherbalmedicineinChina,canimprovethemicrocirculation,increasethetolerancetoischemiainpatientswithmicrocirculatorydisorders.AsCShasbeenfoundtoberenoprotectiveandanti-apoptoticinmultiplekidneyinjuries,wehypothesizedthatCSwouldpreventCIN.TheobjectiveofthisresearchistostudythemechanismofCSontubularepithelialcellapoptosisindiabeticCINrats.

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简介：AbstractBackground:Mesenchymal stem or stromal cells (MSCs) derived from the induced pluripotent stem cells (iPSCs) have uniform biological activity, which makes the clinical application of MSCs in bone repair possible. Culturing the iPSC-MSCs onto osteoconductive materials is a promising tissue engineering-based strategy in bone regeneration. The aim of this work was to evaluate the effects of semaphorin 3A (Sema3A) and hypoxia inducible factor 1 subunit alpha (HIF1α) co-overexpression on the survival and osteogenic differentiation of iPSC-MSCs.Methods:Sema3A and HIF1α were linked together with the three (GGGGS; G, glycine; S, serine) peptide fragment, and their co-expression in iPSC-MSCs was mediated by a lentiviral vector. The fusion protein retained the immune reactivity for both Sema3A and HIF1α as determined with Western blotting. iPSC-MSCs were infected with overexpression lentivirus (oeLenti) as negative control, oeLenti-Sema3A, oeLenti-HIF1α or oeLenti-Sema3A-HIF1α lentiviruses.Results:Sema3A overexpression alone promoted the osteogenic differentiation of iPSC-MSCs (the activity and/or expression of osteoblast markers, such as alkaline phosphatase, osteopontin, and osteocalcin, were upregulated), and suppressed cell survival. The Sema3A-HIF1α fusion protein showed a comparable osteoconductive effect to that of Sema3A without reducing cell survival. We further seeded iPSC-MSCs modified by SemaA-HIF1α overexpression onto hydroxyapatite (HA) scaffolds, and evaluated their growth and differentiation on this three-dimensional material. Additional data indicated that, as compared to iPSC-MSCs cultured in ordinary two-dimensional dishes, cells cultured in HA scaffolds grew (blank vs. HA scaffolds: 0.83 vs. 1.39 for survival) and differentiated better (blank vs. HA scaffolds: 11.29 vs. 16.62 for alkaline phosphatase activity).Conclusion:Modifying iPSC-MSCs with pro-osteogenic (Sema3A) and pro-survival (HIF1α) factors may represent a promising strategy to optimize tissue engineering-based strategy in bone repair.

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简介：ExposureofnaivemurineCD4^+TlymphocytestosuperantigensuchasstaphylococcalenterotoxinB(SEB)inducesastrongproliferativeresponse.ProlongedexposureorsubsequentrestimulationoftherespondingTcellpopulationwithSEBleadstotheapoptoticeventsofactivation-inducedcelldeath(AICD).ThesignalingmechanismresponsiblefortheAICDisatargetofintensiveinvestigation.However,theprecisedownstreamsignahngpathwaysofSEB-inducedAICDremainsunclear.Ourresultshereshowthatthesequentialactivationofcaspase-1/ICE-hkeandcaspase-3/CPP32-hkecysteineproteasesprobablyplaysaroleinthesignalingtransductionofSEB-inducedAICD,butcaspase-3/CPP32-hkeproteasesactivationdoesnotdependoncaspase-1-likeproteasesactivation.HerbimycinA,aspecificinhibitorofproteintyresinekinases,inhibitcaspase-3/CPP32-1ikecysteineproteasesactivation.However,itdoesnotpreventDNAfragmentationofCD4^+TcellsapoptosisinducedbySEB.TheseresultsindicatethatproteintyrosinekinasespathwayisprobablyinvolvedinthesignalingtransductionofCD4^+TcellsapoptosisinducedbySEBand“crosstalks”withthepathwayofcaspase-3/CPP32-1ikeproteasesactivation.

简介：AbstractBackground:MicroRNAs are closely associated with the progression and outcomes of multiple human diseases, including sepsis. In this study, we examined the role of miR-23a in septic injury.MethodsLipopolysaccharide (LPS) was used to induce sepsis in a rat model and H9C2 and HK-2 cells. miR-23a expression was evaluated in rat myocardial and kidney tissues, as well as H9C2 and HK-2 cells. A miR-23a mimic was introduced into cells to identify the role of miR-23a in cell viability, apoptosis, and the secretion of inflammatory cytokines. Furthermore, the effect of Rho-associated kinase 1 (ROCK1), a miR-23a target, on cell damage was evaluated, and molecules involved in the underlying mechanism were identified.Results:In the rat model, miR-23a was poorly expressed in myocardial (sham vs. sepsis 1.00 ± 0.06 vs. 0.27 ± 0.03, P < 0.01) and kidney tissues (sham vs. sepsis 0.27 ± 0.03 vs. 1.00 ± 0.06, P < 0.01). Artificial overexpression of miR-23a resulted in increased proliferative activity (DNA replication rate: Control vs. LPS vs. LPS + Mock vs. LPS + miR-23a: H9C2 cells: 34.13 ± 3.12 vs. 12.94 ± 1.21 vs. 13.31 ± 1.43 vs. 22.94 ± 2.26, P < 0.05; HK-2 cells: 15.17 ± 1.43 vs. 34.52 ± 3.46 vs. 35.19 ± 3.12 vs. 19.87 ± 1.52, P < 0.05), decreased cell apoptosis (Control vs. LPS vs. LPS + Mock vs. LPS + miR-23a: H9C2 cells: 11.39 ± 1.04 vs. 32.57 ± 2.29 vs. 33.08 ± 3.12 vs. 21.63 ± 2.35, P < 0.05; HK-2 cells: 15.17 ± 1.43 vs. 34.52 ± 3.46 vs. 35.19 ± 3.12 vs. 19.87 ± 1.52, P < 0.05), and decreased production of inflammatory cytokines, including interleukin-6 (Control vs. LPS vs. LPS + Mock vs. LPS + miR-23a: H9C2 cells: 59.61 ± 5.14 vs. 113.54 ± 12.30 vs. 116.51 ± 10.69 vs. 87.69 ± 2.97 ng/mL; P < 0.05, F = 12.67, HK-2 cells: 68.12 ± 6.44 vs. 139.65 ± 16.62 vs. 143.51 ± 13.64 vs. 100.82 ± 9.74 ng/mL, P < 0.05, F = 9.83) and tumor necrosis factor-α (Control vs. LPS vs. LPS + Mock vs. LPS + miR-23a: H9C2 cells: 103.20 ± 10.31 vs. 169.67 ± 18.84 vs. 173.61 ± 15.91 vs. 133.36 ± 12.32 ng/mL, P < 0.05, F = 12.67, HK-2 cells: 132.51 ± 13.37 vs. 187.47 ± 16.74 vs. 143.51 ± 13.64 vs. 155.79 ± 15.31 ng/mL, P < 0.05, F = 9.83) in cells. However, ROCK1 was identified as a miR-23a target, and further up-regulation of ROCK1 mitigated the protective function of miR-23a in LPS-treated H9C2 and HK-2 cells. Moreover, ROCK1 suppressed sirtuin-1 (SIRT1) expression to promote the phosphorylation of nuclear factor-kappa B (NF-κB) p65, indicating the possible involvement of this signaling pathway in miR-23a-mediated events.Conclusion:Our results indicate that miR-23a could suppress LPS-induced cell damage and inflammatory cytokine secretion by binding to ROCK1, mediated through the potential participation of the SIRT1/NF-κB signaling pathway.

简介：AbstractObjective:This study aimed at investigating the expression of nuclear factor kappa B (NF-κB) and mammalian target of rapamycin (mTOR) related signal pathways in liver tissues of intrahepatic cholestasis of pregnancy animal models.Methods:Estrogen (EE)-induced cholestasis and a placental ischemia-reperfusion (IR) model were established in pregnant rats. All pregnant rats were divided into four groups by random number table: EE-IR group (n= 6), EE-sham group (n = 6), control-IR group (n= 6) and control-sham group (n= 6). Liver expression of mTOR, its upstream regulator DNA damage response-1 (REDD1), and downstream factor glucose transporter type-1 (GLUT1), accompanied by NF-κB (p65 is the most important component), its activator toll-like receptor 4 (TLR4), and inhibitor IκBα, were detected by western blot analysis and real-time polymerase chain reaction. The intergroup comparisons were performed with a one-way analysis of variance, the comparisons among groups were analyzed with the nonparametric Kruskal-Wallis test.Results:Giving pregnant rats EE alone reduced the hepatic expression of IκBα (0.72 ± 0.20 vs. 1.01 ± 0.07, P= 0.008). Meanwhile, giving pregnant rats placental IR alone increased liver levels of REDD1 (3.24 ± 0.98 vs. 1.06 ± 0.24, P= 0.025), GLUT1 (2.37 ± 0.82 vs. 1.09 ± 0.10, P= 0.039), TLR4 (2.12 ± 0.29 vs. 1.20 ± 0.28, P= 0.010), and p65 (2.09 ± 0.85 vs. 1.04 ± 0.06, P= 0.023), and decreased hepatic mTOR (0.50 ± 0.07 vs. 1.01 ± 0.03, P= 0.001) and IκBα (0.61 ± 0.08 vs. 1.01 ± 0.07, P= 0.014) expression. Subjecting EE-treated rats to placental IR did not further alter liver levels of GLUT1 (2.02 ± 0.45 vs. 1.79 ± 0.39, P= 0.240), TLR4 (2.10 ± 0.74 vs. 1.60 ± 0.36, P= 0.129), or p65 (2.41 ± 0.83 vs. 1.65 ± 0.46, P= 0.145), whereas it did decrease hepatic mTOR (0.42 ± 0.09 vs. 0.90 ± 0.14, P= 0.008) and IκBα (0.43 ± 0.09 vs. 0.72 ± 0.20, P= 0.004) expression and enhance REDD1 expression (4.46 ± 0.65 vs. 2.05 ± 0.47, P= 0.009). Placental IR stress did impact the hepatic expression of REDD1-mTOR-GLUT1 and TLR4/NF-κB/IκBα in pregnant rats.Conclusion:Placental IR-induced hepatic GLUT1, TLR4, and p65 alternation, which responded efficiently in control rats, were impaired in EE-induced ICP rats.