Objective To investigate whether 2,3,5,4'-tetrahydroxystilbene-2-O-β-glucoside(TSG)ameliorated polycystic ovary syndrome(PCOS)-like characteristics by inhibiting inflammation.Methods PCOS models were established ...Objective To investigate whether 2,3,5,4'-tetrahydroxystilbene-2-O-β-glucoside(TSG)ameliorated polycystic ovary syndrome(PCOS)-like characteristics by inhibiting inflammation.Methods PCOS models were established by injecting subcutaneously with dehydroepiandrosterone into female Sprague-Dawley rats,followed by receiving intraperitoneal injection of TSG.The granular cells(GCs)KGN were transfected with small interfering RNAs(si-NC and si-CYP19A1).The cells were preincubated with lipopolysaccharide(LPS)and then treated with or without TSG.The estrous cycle was monitored using vaginal exfoliated cells.The morphology of ovarian follicles was analyzed by H&E staining.ELISA was used to analyze estradiol(E2),testosterone(T),follicle stimulating hormone(FSH),luteinizing hormone(LH),IL-6,TNF-α,AGEs,CRP and Omentin-1 levels in serum.Immunohistochemistry was performed to analyze PCNA and CYP19A1 expressions in the GCs of ovaries.Tunel staining was executed to detect the apoptosis of GCs.Quantitative polymerase chain reaction(qPCR)and Western blot were implemented to measure the expression of CYP19A1 in the ovaries and transfected cells.qPCR was used to analyze the expression of IL-6 and TNF-αin the transfected cells treated with LPS and TSG.Results The estrous cycles were restored in TSG group.Compared with model group,the sinus follicles were reduced and corpus luteums were increased in TSG group.TSG group showed increased E2,and decreased T and LH,compared with model group.Pro-inflammatory factors(IL-6,TNF-α,CRP and AGEs)were decreased,and anti-inflammatory factor(Omentin-1)was increased in TSG group compared with those in model group.TSG could partially inhibit decrease of PNCA-positive GCs and increase of Tunel-positive GCs caused by PCOS.The CYP19A1 expression of GCs in TSG group was upregulated compared with model group.The expressions of IL-6 and TNFαin si-CYP19A1 cells were increased compared with si-NC cells.Compared with cells(si-NC and si-CYP19A1)treated without LPS,the expressions of IL-6 and TNF-αcells were increased,and the expression of CYP19A1 was downregulated in LPS-preincubated cells.Compared with cells treated with LPS,the expression of IL-6 and TNF-αwere decreased,and the expression of CYP19A1 was increased in cells treated with LPS and TSG.Compared with si-NC cells treated with LPS and TSG,the expressions of IL-6 and TNF-αcells were increased in the si-CYP19A1 cells treated with LPS and TSG.Conclusion TSG could alleviate PCOS-like characteristics by increasing the expression of CYP19A1 in GCs to inhibit inflammatory response.展开更多
The capability of recombinant human interleukin-2 ( rhIL-2) and lymphokine-activated killer (LAK) cells in the purging of normal human bone marrows contaminated with human myeloid leukemic cell lines was evaluated. Mi...The capability of recombinant human interleukin-2 ( rhIL-2) and lymphokine-activated killer (LAK) cells in the purging of normal human bone marrows contaminated with human myeloid leukemic cell lines was evaluated. Mixtures of normal human bone marrow mononuclear cells ( BMC) and K562 cells or HL-60 cells (at the BMCK562 ratio of 200:1, 100:1 or 20:1) were incubated with IL-2 with or without LAK cells at the BMC:LAK ratio of 1:1 for one or three days. The nubmers of residual K562 cells, BFU-E and CFU-GM were examined by clonogenic assays. In 200:1 mixture groups without LAK cells, the number of K562 colonies reduced by 50% with no loss of BFU-E and CFU-GM in one-day cultures, and no K562 colonies formed in three-day cultures with about 20% loss of BFU-E and CFU-GM. If the BMC.K562 ratios were 100:1 or 20:1 in the mktures, the leukemic cells could not be eliminated. When the mixtures were incubated with IL-2 and LAK cells, no leukemic cell colonies were detected in the 20:1 group following one-day展开更多
OBJECTIVE To investigate the role and mechanism of G protein-coupled receptor kinase 2(GRK2)involving in hepatocel ular carcinoma(HCC)progression.METHODS Cel Counting Kit 8 and tumor colony formation assay were design...OBJECTIVE To investigate the role and mechanism of G protein-coupled receptor kinase 2(GRK2)involving in hepatocel ular carcinoma(HCC)progression.METHODS Cel Counting Kit 8 and tumor colony formation assay were designed to detect HCC cell proliferation,wound healing assay was to detect HCC migration.The correlation between GRK2 and early growth response-1(EGR1)were detected by RT-PCR and real-time PCR assays.Co-immunoprecipitation and Western blot assay were adopted to detect the relationship between GRK2and insulin-like growth factor 1 receptor(IGF-1R)signaling pathway.RESULTS In this study we find that GRK2plays an inhibition role in IGF1-induced HCC cell proliferation and migration.Overexpression of GRK2 causes a decrease in EGR1 expression,while knockdown of GRK2 leads to the dramatically increase in EGR1 expression in the treatment of IGF1.Through co-immunoprecipitation and Western blot assay,we confirm that GRK2can interact with IGF-1R and inhibiting IGF1-induced activation of IGF1R signaling pathway.Silencing EGR1attenuates GRK2 overexpression-caused inhibition of cell proliferation,tumor colony number and migrationactivity,while overexpressing of EGR1 restores the antiproliferative and migratory effect by GRK2 overexpression in HCCLM3 cells.CONCLUSION Taken together,these results suggest that GRK2 may inhibit IGF1-induced HCC cell growth and migration through down-regulation of EGR1 and indicate that enforced GRK2 may offer a potential therapeutic approach against HCC.展开更多
Ami To investigate PGE2 and TNF-alpha signaling pathway involving in the maturation and activation of bone marrow dendritic cells （DCs） and the effect of CP-25. Method Bone marrow DCs were isolated and stim- ulated ...Ami To investigate PGE2 and TNF-alpha signaling pathway involving in the maturation and activation of bone marrow dendritic cells （DCs） and the effect of CP-25. Method Bone marrow DCs were isolated and stim- ulated by PGE2 and TNF-alpha respectively. The markers of maturation and activation expressed on DCs, such as CD40, CD80, CD83, CD86, MHC-II, and the ability of antigen uptake of DCs were analyzed by flow cytometry. The proliferation of T cells co-cultured with DCs, the signaling pathways of PGE2-EP4-cAMP and TNF-alpha- TRADD-TRAF2-NF-KB in DCs were analyzed. Result Both PGE2 and TNF-alpha up-regulated the expressions of CD40, CD80, CD83, CD86, and MHC-II, decreased the antigen uptake of DCs, and DCs stimulated by PGE2 or TNF-alpha could increase T cell proliferation. CP-25 （ 10 -5, 10 -6 , 10 -7 mol/L ） decreased significantly the ex- pressions of CD40, CD80, CD83, CD86 and MHC- ]I , increased the antigen uptake of DCs, and suppressed T cell proliferation induced by DCs. PGE2 increased the expressions of EP4, NF-KB and down-regulated cAMP level of DCs. TNF-alpha could also up-regulate TNFR1, TRADD, TRAF2, and NF-KB expression of DCs. CP-25 （10^-5, 10^-6, 10^-7 mol/L） decreased the expressions of EP4 and NF-KB, increased cAMP level in DCs stimulated by PGE2. CP-25 （10^-5 10^-6 10^-7 mol/L） also could down-regulate significantly TNFR1 TRADD TRAF2 and NF-KB expression in DCs stimulated by TNF-alpha. Conclusion PGE2 and TNF-alpha could enhance DCs func- tions by mediating PGE2-EP4-cAMP pathway, TNF-alpha-TNFR1-TRADD-TRAF2-NF-KB pathway respectively. CP-25 might inhibit the function of DCs through regulating PGE2-EP4-cAMP and TNF-alpha-TNFR1-TRADD- TRAF2-NF-KB pathways.展开更多
OBJECTIVE The present study was designed to investigate anticancer effect of zeylenone(Zey)on K562 cells derived from chronic myelogenous leukemia(CML)both in vitro and in vivo,followed by exploring the underlying mec...OBJECTIVE The present study was designed to investigate anticancer effect of zeylenone(Zey)on K562 cells derived from chronic myelogenous leukemia(CML)both in vitro and in vivo,followed by exploring the underlying mechanisms.METHODS Initially,the effects of Zey on cel viability,proliferation,and apoptosis were measured in K562 cells by MTT,soft agar assay,AO/EB staining,hoechst 33258 staining and flow cytometric analysis after they were treated with Zey for indicated time,the involving signaling pathways were then investigated by JC-1,real-time quantitative polymerase chain reaction(RT-q PCR),Western blotting and immunofluorescence analysis.Furthermore,the in vivo anti-tumoractivity of Zey was assessed with nude xenografts and the involving mechanism was confirmed by immunohistochemical(IHC)and histopathological analysis.RESULTS We identified that Zey dose-dependently decreased cell viability,colony formation and expression of Proliferating Cell Nuclear Antigen(PCNA),and significantly induced K562 cell apoptosis via regulating Bcl-2 family members,decreasing mitochondrial transmembrane potential,and activating caspase-3,caspase-9,and caspase-8(P<0.05 or P<0.01).Further study revealed that Zey significantly inhibited phosphorylation of Jak2 and Src and downregulated their downstream proteins,including stat3,PI3K/AKT/m TOR,and ERK1/2 signaling pathways(P<0.05 or P<0.01).Zey also suppressed tumor growth with low toxicity in mouse xenograft model of K562cells through decreasing expression of Jak2 and Src.CONCLUSION Our data demonstrated that Zey substantially suppressed K562 cells both in vitro and in vivo through Jak2 and Src pathways.These findings suggest the potential of Zey as an effective anticancer agent in CML treatment.展开更多
Manganese(Mn),an essential trace element in the human body,plays critical roles in many biological processes.Recent studies have discovered that Mn^(2+)may promote or directly activate the cGAS-STING pathway,thereby s...Manganese(Mn),an essential trace element in the human body,plays critical roles in many biological processes.Recent studies have discovered that Mn^(2+)may promote or directly activate the cGAS-STING pathway,thereby subsequently initiating the natural immune response and augmenting antitumor therapy.However,the current lack of accurate methods for Mn^(2+)determination in cells significantly limits their mechanism investigation;hence,it is urgent to establish novel tools to detect Mn^(2+)in cells.In this study,the dual-emission carbon dots were initially synthesized via the one-pot hydrothermal method employing L-aspartic acid and p-phenylenediamine as raw materials.In the presence of Mn^(2+),the emission peak centered at 350 nm exhibited significant enhancement,whereas another peak at 610 nm remained stable.Consequently,a ratiometric sensor for Mn^(2+)determination was established using the signal at 350 nm as the responsive signal and the signal at 610 nm as an internal reference.Under the optimal condition,a good linear relationship was achieved between the F350/F610 value and Mn^(2+)concentration ranging from 0.9 to 15μmol/L,with a calculated LOD of 61 nmol/L.Benefiting from the special Mn^(2+)-induced ratiometric approach,this method demonstrates outstanding sensitivity,selectivity,and stability,rendering it applicable for Mn^(2+)determination in complex biological samples,as well as Mn^(2+)imaging in MKN-45 and LO2 cells.展开更多
Gastrin and cyclooxygenase-2(COX-2) playimportant roles in the carcinogenesis and progression ofgastric cancer.However,it remains unknown whether the combination of cholecystokinin-2(CCK-2) receptor antagonist plus CO...Gastrin and cyclooxygenase-2(COX-2) playimportant roles in the carcinogenesis and progression ofgastric cancer.However,it remains unknown whether the combination of cholecystokinin-2(CCK-2) receptor antagonist plus COX-2 inhibitor exerts synergistic anti-tumor effects on human gastric cancer.Here,we demonstrated that the combination of AG-041R(a CCK-2 receptor antagonist) plus NS-398(a selective COX-2 inhibitor) treatment had synergistic effects on proliferation inhibition,apoptosis induction,down-regulation of Bcl-2 and up-regulation of Bax expression in MKN-45 cells.These results indicate that simultaneous targeting of CCK-2 receptor and COX-2 may inhibit gastric cancer development more effectively than targeting either molecule alone.(C)2008 Elsevier Ireland Ltd.All rights reserved.展开更多
Aim The present study developed a CYP3A4-expressed Caco-2 monolayer model at which effects of the efflux-metabolism alliance on the transport and uptake of clausenamide（CLA） enantiomers as CYP3A4 substrates were inv...Aim The present study developed a CYP3A4-expressed Caco-2 monolayer model at which effects of the efflux-metabolism alliance on the transport and uptake of clausenamide（CLA） enantiomers as CYP3A4 substrates were investigated. The apparent permeability coefficients （Papp） of （ - ） and （ ＋ ）CLA were higher in the ab- sorptive direction than those in the secretory direction with efflux ratios（ER） of 0. 709 ± 0.411 and 0. 867± 0. 250 （ Х10^-6 -1 cm · s ）, respectively. Their bidirectional transports were significantly reduced by （75.6 ± 87.5）% af- ter treatment with verapamil （ a P-glycoprotein inhibitor） that increased the rate of metabolism by CYP3 A4, whereas the CYP3A4 inhibitor ketoconazole treatment markedly enhanced the basolateral to apical flux of （ - ） and （ ＋ ） CLA with ERs being 2. 934 ± 1. 432 and 1. 877 ± 0. 148 （ Х 10^-6 cm/s） respectively. These changes could be blocked by the duel CYP3A4/P-glycoprotein inhibitor cyclosporine A, consequently, Papp values for CLA enanti- omers in both directions were significantly greater than those obtained by using verapamil or ketoconazole, and their ERs were similar to those following （ - ） or （ ＋ ）-isomer treatment alone. Furthermore, the uptake of （ - ）CLA was more than that of （ ＋ ）CLA in the transfected cells. Incubation with ketoeonazole decreased the intracellular concentrations of the two enantiomers. This effect disappeared in the presence of a CYP3A4 inducer dexametha- sone. These results indicated that CYP3A4 could influence P-gp efflux, transport and uptake of CLA enantiomers as CYP3A4 substrates and that a duel inhibition to CYP3A4/ P-glycoprotein could enhance their absorption and bioavailability, which provides new insight into the efflux-metabolism alliance and will benefit the clinical pharma- cology of （？） CLA as a candidate drug for treatment of Alzheimer＇ s disease.展开更多
Anatase Ti0_(2) nanosheet-based hierarchical spheres(HSs)with nearly 100%exposed{001}facets were synthesized via a facile solvothermal process.Using these hierarchical spheres as a scattering layer on nanocrystaline T...Anatase Ti0_(2) nanosheet-based hierarchical spheres(HSs)with nearly 100%exposed{001}facets were synthesized via a facile solvothermal process.Using these hierarchical spheres as a scattering layer on nanocrystaline TiO_(2)film,hi-layered dye-sensitized solar cells(DSSCs)have been fabricated by electrophoresis deposition method,which well preserved the fragile hierarchical structure.Owing to the superior dye adsorption and light scattering effect of HSs,an overall energy conversion efficiency of 7.38%is achieved,which is 26%higher than that of nanoparticle-based photoanode.展开更多
Based on published sequences for chicken Bcl-2,three siRNAs（small interfering RNA）were designed,and expression vectors were constructed and transfected into goose granulosa cells cultured in vitro.Bcl-2 protein,apop...Based on published sequences for chicken Bcl-2,three siRNAs（small interfering RNA）were designed,and expression vectors were constructed and transfected into goose granulosa cells cultured in vitro.Bcl-2 protein,apoptosis and proliferation of granulosa cells,48 h after the transf ection,were analyzed by flow cytometry,and progesterone（P）secreted into the culture medium was measured by radioimmunoassay.In addition,apoptosis and Bcl-2 protein level were assessed in untreated granulosa cells from the four largest preovulatory follicles（F<sub>1</sub><sup>F</sup><sub>4</sub>）,the smallest preovulatory follicles（SPF）,small yellow follicles（SYF）and atretic follicles.The highest level of Bcl-2 protein was observed in granulosa cells from SPF,and levels in cells from healthy follicles were significantly higher than those of atretic follicles（P【0.05）.Bcl-2 protein levels in cells subjected to RNAi were significantly lower than those of controls（P【0.05）,while apoptosis indices（AI）,proliferation indices（PI）and P secretion in the RNAi treatments were higher than those of controls（P【0.05）.展开更多
OBJECTIVE Liguzinediol is a derivative of the natural active ingredient ligustrazine,and we found that liguzinediol has significant positive inotropic effects,which are stronger than that of TMP.Besides,it does not le...OBJECTIVE Liguzinediol is a derivative of the natural active ingredient ligustrazine,and we found that liguzinediol has significant positive inotropic effects,which are stronger than that of TMP.Besides,it does not lead to arrhythmia,hypotension and other side effects.This study aims to investigate the anti-apoptotic effects of liguzinediolon H9C2 cells.METHODS Apoptotic H9C2 cells induced by DOX were observed by electron microscope and FCM analysis.The protein expressions of Bax,Bcl-2,caspases 3 and NF-κB were detected by Western blotting.RESULTS Apoptotic H9C2 cells induced by DOX were observed,but without apoptotic bodies in liguzinediol group.Declined peak of H9C2 cell apoptosis was seen in liguzinediol group by FCM analysis.And downregulation of Bax,caspases 3,NF-κB and upregulation of Bcl-2 were found by Western blotting.CONCLUSION Liguzinediol protected cardiomyocytes against apoptosis through downregulation of Bax and caspases 3 and upregulation of Bcl-2.Liguzinediol can inhibited cardiomyocyte apoptosis through the NF-κB signal pathway.展开更多
OBJECTIVE Hepatocel ular carcinoma(HCC)is the most common cause of cancer-related mortality,with high incidence rates,robust metastatic propensity and acquired resistance to therapy.Metformin,an extensively prescribed...OBJECTIVE Hepatocel ular carcinoma(HCC)is the most common cause of cancer-related mortality,with high incidence rates,robust metastatic propensity and acquired resistance to therapy.Metformin,an extensively prescribed and well-tolerated first-linetherapeutic drug for type 2 diabetes mellitus,has recently been identified as a potential and attractive anticancer adjuvant drug combined with chemotherapeutics to improve treatment efficacy and lower doses.Curcumin,a botanical extracts,has been shown antitumorigenic properties.This study aims to investigate the combinational effect of metformin and curcumin on inbibition of tumor growth and metastasis in Hep G2 cells and the possible underlying mechanisms.METHODS The cell proliferation was determined by MTT,CCK-8 and colony formation assay.The protein expression was detected by Western blotting.Activity of MMP-2 and MMP-9 was estimated by gelatin zymography.Flow cytometry analysis was used to evaluate the influence of metformin and curcumin on cell cycle arrest and apoptosis,and morphology observation of apoptosis was detected by Hoechst33342.Scratch and transwell assay was performed to detect the cell migration and invasion.The suppression of this combination therapy oncapillary tube formation was detected by tube formation assay.RESULTS Combination of metformin and curcumin induced stronger inhibition on Hep G2 cells proliferation than monotherapywhich related to induction of cell cycle arrest in G2/M phase and apoptosis through regulation of the protein expression of cyclin B and Bcl-2/Bax.Moreover,the co-treatment of metformin with curcumin exerted an enhanced inhibitory effect on Hep G2 cell metastasis and synergistically inhibited the tube formation of HUVEC cells.The suppression of PI3K/AKT/m TOR pathway and inhibition the protein expression of STAT3,MMP9,MMP2 and VEGF might involve in this synergistic effects of combination treatment.CONCLUSION Combination of metformin and curcumin inhibited Hep G2 cells proliferationmore effectively than monotherapy and synergistically induced a greater inhibition on migration and invasion of Hep G2 cells.展开更多
OBJECTIVE To investigate the effects of ex-tract of Ramulus Cinnamom(RC)against LPS-induced inflammation in microglia.METHODS Activated microglia releases various pro-inflammatory cytokines to induce neuroinflammation...OBJECTIVE To investigate the effects of ex-tract of Ramulus Cinnamom(RC)against LPS-induced inflammation in microglia.METHODS Activated microglia releases various pro-inflammatory cytokines to induce neuroinflammation in stroke.Lipopolysaccaride(LPS)is an endotoxin from the outer membrane of Gram-negative bacteria that activates microglia.MTT assay was used to observe the cell viability.The content of NO in supernatant was measured by Griess reagent.The levels of IL-1β,IL-6 and TNF-αin supernatant were detected by ELISA kits.The intracellular COX-2,TLR4,and My D88expression was assayed by Western blotting.RESULTS RC extract 30 and 100μg·m L-1significantly decreased the production of related inflammatory factors such as NO(P<0.05,P<0.01),IL-1β(P<0.01,P<0.01),IL-6(P<0.05,P<0.01)and TNF-α(P<0.01,P<0.01).Furthermore,RC extract significantly inhibited the COX-2,TLR4,and My D88 expression induced by LPS in BV2cells.CONCLUSION RC extract may have therapeutic potential for the improvement of neuroinflammation,and the mechanism may be involved in down-regulation of TLR4/My D88 inflammation pathway.展开更多
OBJECTIVE Aconitine(ACO)as the main active component in Aconitum carmichaelii debeaux(family Ranunlaceae),has highly toxicity in heart and the mechanisms are not clear yet.Paeoniflorin(PF),the main chemical ingredient...OBJECTIVE Aconitine(ACO)as the main active component in Aconitum carmichaelii debeaux(family Ranunlaceae),has highly toxicity in heart and the mechanisms are not clear yet.Paeoniflorin(PF),the main chemical ingredient in Herbaceous peony,can protect heart hurt by antioxidant,vasodilator effect and other effects.In this study,we focused on investigating the mechanism of PF reducing the cardiotoxicity of ACO.METHODS We chose H9c2 cells as experimental subject.MTT,Western blotting and real-time PCR were used to measure cell proliferation,apoptosis,ion channels and oxidative stress.RESULTS Cell proliferation in ACO+PF group was significantly increased compared with ACO group;the ratio with Bcl-2 and Bax and the level of p53 were upregulated by PF,while the level of caspase-3 was lightly reduced.The expression of SCN5A mRNA significantly was increased in ACO+PF group,while the expres⁃sion of RyR2 and Cx43 mRNA was dropped.Compared with ACO group,extracellular LDH and intracellular MDA were highly decreased,while intracellular SOD was regulated.CONCLUSION Cardiotoxicity of ACO in H9c2 cells was signifi⁃cantly decreased by PF.展开更多
OBJECTIVE To investigate the effect of P2X7receptor(P2X7r)inhibition,using a specific inhibitor(A438079)to prevent the development of liver fibrosis on human hepatic stellate cells,LX-2.METHODS The supernatant from li...OBJECTIVE To investigate the effect of P2X7receptor(P2X7r)inhibition,using a specific inhibitor(A438079)to prevent the development of liver fibrosis on human hepatic stellate cells,LX-2.METHODS The supernatant from lipopolysaccharide(LPS)-stimulated RAW264.7 mouse macrophages was supplemented to LX-2 cells for 24 h.LX-2cells were primed with LPS for 4h and subsequently stimulated for 30 min with 3mmol·L-1 of adenosine 5′-triphosphate(ATP).A438079(10μmol·L-1)was supplemented to LX-2 cells 10 min prior to ATP.RESULTS Directly treated with LPS on LX-2 cells,mRNA expressions of IL-1β,IL-18 and IL-6 were increased,as well as P2X7 r.And caspase-1,ASC and NLRP3 mRNA expressions were increased with LPS stimulation.LPS stimulation also increasedα-SMA and collagenⅠ mRNA expressions.Interestingly treatment of LX-2cells with mediums from LPS-primed RAW264.7mouse macrophages exhibited greater increase of mRNA expressions of above genes than those in LX-2directly treated with LPS.Pretreatment of directly or indirectly LPS-stimulated LX-2 cells with A438079 both suppressed IL-1βmRNA expression.In addition treatment of LPS-primed LX-2 cells with 3mmol·L-1 ATP induced the significant increase of IL-1β,IL-6,caspase-1,pannexin-1,α-SMA and collagenⅠ mRNA expression,the increasing ofα-SMA protein expression and cleavage of IL-1β.These events were significantly suppressed by pretreatment with P2X7 rantagonist A438079.P2X7 rblockade also significantly reduced the protein expression ofα-SMA.CONCLUSION Our results suggest that the involvement of the P2X7r-NLRP3 inflammasome pathway in the secretion of IL-1βfrom extracellular ATP/LPS-stimulated human hepatic stellate cells.This study demonstrated that repression of the P2X7 rrepresents a novel potential therapeutic approach to control liver fibrosis.展开更多
OBJECTIVE A novel mast cell-specific G-protein-coupled receptor(GPCR),known as mas-related GPCR-B2(MRGPRB2),plays important roles in the immune response.The opening of ion channels mediated by MRGPRB2 activation remai...OBJECTIVE A novel mast cell-specific G-protein-coupled receptor(GPCR),known as mas-related GPCR-B2(MRGPRB2),plays important roles in the immune response.The opening of ion channels mediated by MRGPRB2 activation remains unclear.METHODS AND RE⁃SULTS Calcium influx induced by activation of MRGPRB2 receptor in mouse peritoneal mast cells was related to the concentrations of calcium ions in the extracellular solution.Similarly,the volt⁃age-dependent current generated by MRGPRB2 activation was also correlated with the extracellu⁃lar calcium concentration.In addition,the in⁃creased of calcium influx or voltage-dependent current caused by activation of MrgprB2 could be blocked by U73122(PLC blocker)or 2-APB(IP3-ORAI1 blocker).Meanwhile,calcium-activated chlorine channel(TMEM16A)was involved in the generation of voltage-dependent currents in⁃duced by MRGPRB2 activation in mouse perito⁃neal mast cells.Furthermore,the degranulation of mouse peritoneal mast cells mediated by MRGPRB2 receptor could also be inhibited by U73122 or 2-APB.CONCLUSION PLC-IP3-ORAI1-TMEM16A signaling pathway was involved in MRGPRB2-mediated mast cell activation.展开更多
P2X7 receptor （P2X7r） is important in inflammation and fibrosis. The aim of the present study was to investigate the effect of P2X7r inhibition, using a specific inhibitor （A438079） to prevent the development of l...P2X7 receptor （P2X7r） is important in inflammation and fibrosis. The aim of the present study was to investigate the effect of P2X7r inhibition, using a specific inhibitor （A438079） to prevent the development of liver fibrosis on human hepatic stellate cells, LX-2. The supernatant from lipopolysaccharide （LPS）-stimulated RAW 264.7 mouse macrophages was supplemented to LX-2 cells for 24 h. LX-2 cells were primed with LPS for 4 h and subsequently stimulated for 30 rain with 3 mmol · L^-1 of adenosine 5＇-triphosphate （ATP）. A438079 （ 10 μmol · L^-1） was supplemented to LX-2 cells 10 rain prior to ATP. Directly treated with LPS on LX-2 cells, mRNA ex- pressions of IL-1β, IL-18 and IL-6 were increased, as well as P2X7r. And caspase-1, ASC and NLRP3 mRNA ex- pressions were increased with LPS stimulation. LPS stimulation also increased oL-SMA and collagen I mRNA expres- sions. Interestingly treatment of LX-2 cells with mediums from LPS-primed RAW 264.7 mouse macrophages exhibi- ted greater increase of mRNA expressions of above genes than those in LX-2 directly treated with LPS. Pretreatment of directly or indirectly LPS-stimulated LX-2 cells with A438079 both suppressed IL-1β mRNA expression. In addi- tion treatment of LPS-primed LX-2 cells with 3 mM ATP induced the significant increase of IL-1β, IL-6, caspase- 1, pannexin-1, α-SMA and collagen I mRNA expression, the increasing of oL-SMA protein expression and cleavage of IL-1β. These events were significantly suppressed by pretreatment with P2X7r antagonist A438079. P2XTr blockade also significantly reduced the protein expression of oL-SMA. Our results suggest that the involvement of the P2X7r-NLRP3 inflammasome pathway in the secretion of IL-1β from extracellular ATP/LPS-stimulated human he- patic stellate cells. This study demonstrated that repression of the P2XTr represents a novel potential therapeutic ap- proach to control liver fibrosis.展开更多
Significant progress has recently been made in enhancing the power conversion efficiency(PCE)of perovskite solar cells(PSCs).The electron transport layer(ETL),as an essential component of PSCs,significantly influences...Significant progress has recently been made in enhancing the power conversion efficiency(PCE)of perovskite solar cells(PSCs).The electron transport layer(ETL),as an essential component of PSCs,significantly influences the performance of devices.Traditional spin-coating method for preparing the ETL fails to fully cover the cusp of FTO transparent conductive glass substrate,leading to direct contact between perovskite film and FTO substrate,which induces charge recombination and reduces the performance of PSCs.To address this issue,an in-situ growth method was proposed to prepare conformal SnO_(2) films on FTO glass substrates in this study.The resulting SnO_(2) films are not only dense and uniform,fully covering the cusp of the FTO glass substrates and reducing the contact area between the FTO substrates and the perovskite films,but also facilitating the formation of perovskite films with large grain sizes.Moreover,the conformal SnO_(2) films can improve the charge extraction at the SnO_(2)/perovskite interface,reduce the trap density and trap-assisted recombination in PSCs,and thus enhance the PCE of PSCs.Through comparative experiments,it is found that the PSCs with in-situ grown SnO_(2) films show an improved PCE of 21.97%,which significantly increased compared to that with spin-coated SnO_(2) films(20.93%).All above data demonstrate that the as-prepared SnO_(2) film can serve as an ideal ETL.It is worth mentioning that this method avoids the use of corrosive hydrochloric acid and toxic thioglycolic acid,and it can also be extended to ITO flexible transparent conductive substrates in the future.展开更多
Porcine epidemic diarrhea(PED)is caused by porcine epidemic diarrhea virus(PEDV),and is characterized by vomiting,diarrhea and dehydration of suckling pigs from 80% to 100% morbidity and 50% to 90% mortality,and resul...Porcine epidemic diarrhea(PED)is caused by porcine epidemic diarrhea virus(PEDV),and is characterized by vomiting,diarrhea and dehydration of suckling pigs from 80% to 100% morbidity and 50% to 90% mortality,and resulted in tremendous economic losses to swine industry.The PEDV mainly infects small intestine of pigs,resulting in vacuolar degeneration and necrosis of mucosal epithelium.The IPEC-J2 is a pig intestine epithelial cell line,which is similar to the intestinal environment of piglets,can be used to isolate and identify the PEDV field isolates.In this study,it appeared the PEDV typical postmortem changes and histopathological lesion of degeneration and destruction of small intestine in infected piglets,and IHC identified that the PEDV distributed in the mucosa and submucosa of small intestine mostly.Furthermore,the PEDV HLJ strain was successfully isolated and characterized in the IPEC-J2 cells,and indicated that the IPEC-J2 cell line was sensitive to isolate and adapt the PEDV field strain,and could be utilized to multiply the PEDV rapidly.The S gene analysis indicated that the PEDV HLJ strain was the prevailed virus,belonged to Group 1 with attenuated virulent DR13,SC1402 and J-S2/2015 strains isolated in South Korea and China from 2014 to 2015.This study had important theoretical and practical significances on analyzing genetic variation of the PEDV,understanding the pathogenic characteristics of the virus and developing new vaccines for the PED.展开更多
MoS_(2)/CuS composite catalysts were successfully synthesized using a one-step hydrothermal method with sodium molybdate dihydrate,thiourea,oxalic acid,and copper nitrate trihydrate as raw materials.The hydrogen pro-d...MoS_(2)/CuS composite catalysts were successfully synthesized using a one-step hydrothermal method with sodium molybdate dihydrate,thiourea,oxalic acid,and copper nitrate trihydrate as raw materials.The hydrogen pro-duction performance of MoS_(2)/CuS prepared with different molar ratios of Mo to Cu precursors(n_(Mo)∶n_(Cu))as cathodic catalysts was investigated in the two-chamber microbial electrolytic cell(MEC).X-ray diffraction(XRD),X-ray pho-toelectron spectroscopy(XPS),scanning electron microscopy(SEM),transmission electron microscope(TEM),linear scanning voltammetry(LSV),electrochemical impedance analysis(EIS),and cyclic voltammetry(CV)were used to characterize the synthesized catalysts for testing and analyzing the hydrogen-producing performance.The results showed that the hydrogen evolution performance of MoS_(2)/CuS-20%(nMo∶nCu=5∶1)was better than that of platinum(Pt)mesh,and the hydrogen production rate of MoS_(2)/CuS-20%as a cathode in MEC was(0.2031±0.0237)m^(3)_(H_(2))·m^(-3)·d^(-1) for 72 h at an applied voltage of 0.8 V,which was slightly higher than that of Pt mesh of(0.1886±0.0134)m^(3)_(H_(2))·m^(-3)·d^(-1).The addition of a certain amount of CuS not only regulates the electron transfer ability of MoS_(2) but also increases the density of active sites.展开更多
文摘Objective To investigate whether 2,3,5,4'-tetrahydroxystilbene-2-O-β-glucoside(TSG)ameliorated polycystic ovary syndrome(PCOS)-like characteristics by inhibiting inflammation.Methods PCOS models were established by injecting subcutaneously with dehydroepiandrosterone into female Sprague-Dawley rats,followed by receiving intraperitoneal injection of TSG.The granular cells(GCs)KGN were transfected with small interfering RNAs(si-NC and si-CYP19A1).The cells were preincubated with lipopolysaccharide(LPS)and then treated with or without TSG.The estrous cycle was monitored using vaginal exfoliated cells.The morphology of ovarian follicles was analyzed by H&E staining.ELISA was used to analyze estradiol(E2),testosterone(T),follicle stimulating hormone(FSH),luteinizing hormone(LH),IL-6,TNF-α,AGEs,CRP and Omentin-1 levels in serum.Immunohistochemistry was performed to analyze PCNA and CYP19A1 expressions in the GCs of ovaries.Tunel staining was executed to detect the apoptosis of GCs.Quantitative polymerase chain reaction(qPCR)and Western blot were implemented to measure the expression of CYP19A1 in the ovaries and transfected cells.qPCR was used to analyze the expression of IL-6 and TNF-αin the transfected cells treated with LPS and TSG.Results The estrous cycles were restored in TSG group.Compared with model group,the sinus follicles were reduced and corpus luteums were increased in TSG group.TSG group showed increased E2,and decreased T and LH,compared with model group.Pro-inflammatory factors(IL-6,TNF-α,CRP and AGEs)were decreased,and anti-inflammatory factor(Omentin-1)was increased in TSG group compared with those in model group.TSG could partially inhibit decrease of PNCA-positive GCs and increase of Tunel-positive GCs caused by PCOS.The CYP19A1 expression of GCs in TSG group was upregulated compared with model group.The expressions of IL-6 and TNFαin si-CYP19A1 cells were increased compared with si-NC cells.Compared with cells(si-NC and si-CYP19A1)treated without LPS,the expressions of IL-6 and TNF-αcells were increased,and the expression of CYP19A1 was downregulated in LPS-preincubated cells.Compared with cells treated with LPS,the expression of IL-6 and TNF-αwere decreased,and the expression of CYP19A1 was increased in cells treated with LPS and TSG.Compared with si-NC cells treated with LPS and TSG,the expressions of IL-6 and TNF-αcells were increased in the si-CYP19A1 cells treated with LPS and TSG.Conclusion TSG could alleviate PCOS-like characteristics by increasing the expression of CYP19A1 in GCs to inhibit inflammatory response.
文摘The capability of recombinant human interleukin-2 ( rhIL-2) and lymphokine-activated killer (LAK) cells in the purging of normal human bone marrows contaminated with human myeloid leukemic cell lines was evaluated. Mixtures of normal human bone marrow mononuclear cells ( BMC) and K562 cells or HL-60 cells (at the BMCK562 ratio of 200:1, 100:1 or 20:1) were incubated with IL-2 with or without LAK cells at the BMC:LAK ratio of 1:1 for one or three days. The nubmers of residual K562 cells, BFU-E and CFU-GM were examined by clonogenic assays. In 200:1 mixture groups without LAK cells, the number of K562 colonies reduced by 50% with no loss of BFU-E and CFU-GM in one-day cultures, and no K562 colonies formed in three-day cultures with about 20% loss of BFU-E and CFU-GM. If the BMC.K562 ratios were 100:1 or 20:1 in the mktures, the leukemic cells could not be eliminated. When the mixtures were incubated with IL-2 and LAK cells, no leukemic cell colonies were detected in the 20:1 group following one-day
基金The project supported by National Natural Science Foundation of China(81502123,81330081,81202596)Natural Science Foundation of Anhui Province(1308085QH130)+3 种基金Anhui Province Natural Science Foundation in University(KJ2014A119)Grants for Scientific Research of BSKY from Anhui Medical University(XJ201212)Specialized Research Fund for the Doctoral Program of Higher Education(20113420120006,20123420110003)Program for Tackling Key Problems in Science and Technology by Anhui Province(1301042098)
文摘OBJECTIVE To investigate the role and mechanism of G protein-coupled receptor kinase 2(GRK2)involving in hepatocel ular carcinoma(HCC)progression.METHODS Cel Counting Kit 8 and tumor colony formation assay were designed to detect HCC cell proliferation,wound healing assay was to detect HCC migration.The correlation between GRK2 and early growth response-1(EGR1)were detected by RT-PCR and real-time PCR assays.Co-immunoprecipitation and Western blot assay were adopted to detect the relationship between GRK2and insulin-like growth factor 1 receptor(IGF-1R)signaling pathway.RESULTS In this study we find that GRK2plays an inhibition role in IGF1-induced HCC cell proliferation and migration.Overexpression of GRK2 causes a decrease in EGR1 expression,while knockdown of GRK2 leads to the dramatically increase in EGR1 expression in the treatment of IGF1.Through co-immunoprecipitation and Western blot assay,we confirm that GRK2can interact with IGF-1R and inhibiting IGF1-induced activation of IGF1R signaling pathway.Silencing EGR1attenuates GRK2 overexpression-caused inhibition of cell proliferation,tumor colony number and migrationactivity,while overexpressing of EGR1 restores the antiproliferative and migratory effect by GRK2 overexpression in HCCLM3 cells.CONCLUSION Taken together,these results suggest that GRK2 may inhibit IGF1-induced HCC cell growth and migration through down-regulation of EGR1 and indicate that enforced GRK2 may offer a potential therapeutic approach against HCC.
文摘Ami To investigate PGE2 and TNF-alpha signaling pathway involving in the maturation and activation of bone marrow dendritic cells （DCs） and the effect of CP-25. Method Bone marrow DCs were isolated and stim- ulated by PGE2 and TNF-alpha respectively. The markers of maturation and activation expressed on DCs, such as CD40, CD80, CD83, CD86, MHC-II, and the ability of antigen uptake of DCs were analyzed by flow cytometry. The proliferation of T cells co-cultured with DCs, the signaling pathways of PGE2-EP4-cAMP and TNF-alpha- TRADD-TRAF2-NF-KB in DCs were analyzed. Result Both PGE2 and TNF-alpha up-regulated the expressions of CD40, CD80, CD83, CD86, and MHC-II, decreased the antigen uptake of DCs, and DCs stimulated by PGE2 or TNF-alpha could increase T cell proliferation. CP-25 （ 10 -5, 10 -6 , 10 -7 mol/L ） decreased significantly the ex- pressions of CD40, CD80, CD83, CD86 and MHC- ]I , increased the antigen uptake of DCs, and suppressed T cell proliferation induced by DCs. PGE2 increased the expressions of EP4, NF-KB and down-regulated cAMP level of DCs. TNF-alpha could also up-regulate TNFR1, TRADD, TRAF2, and NF-KB expression of DCs. CP-25 （10^-5, 10^-6, 10^-7 mol/L） decreased the expressions of EP4 and NF-KB, increased cAMP level in DCs stimulated by PGE2. CP-25 （10^-5 10^-6 10^-7 mol/L） also could down-regulate significantly TNFR1 TRADD TRAF2 and NF-KB expression in DCs stimulated by TNF-alpha. Conclusion PGE2 and TNF-alpha could enhance DCs func- tions by mediating PGE2-EP4-cAMP pathway, TNF-alpha-TNFR1-TRADD-TRAF2-NF-KB pathway respectively. CP-25 might inhibit the function of DCs through regulating PGE2-EP4-cAMP and TNF-alpha-TNFR1-TRADD- TRAF2-NF-KB pathways.
基金The project supported by PUMC Youth Fund(3332015047)Fundamental Research Funds for the Central Universities,Beijing Key Laboratory of Innovative Drug Discovery of Traditional Chinese Medicine(Natural Medicine)and Translational Medicine,Institute of Medical Plant Development,Peking Union Medical College and Chinese Academy of Medical Sciencesby the National Science and Technology Major Project and Scientific Researchers Aiding Enterprise Item(2012ZX09301-002-001-026 and 2012ZX09501001-004)from the Ministry of Science and Technology of China
文摘OBJECTIVE The present study was designed to investigate anticancer effect of zeylenone(Zey)on K562 cells derived from chronic myelogenous leukemia(CML)both in vitro and in vivo,followed by exploring the underlying mechanisms.METHODS Initially,the effects of Zey on cel viability,proliferation,and apoptosis were measured in K562 cells by MTT,soft agar assay,AO/EB staining,hoechst 33258 staining and flow cytometric analysis after they were treated with Zey for indicated time,the involving signaling pathways were then investigated by JC-1,real-time quantitative polymerase chain reaction(RT-q PCR),Western blotting and immunofluorescence analysis.Furthermore,the in vivo anti-tumoractivity of Zey was assessed with nude xenografts and the involving mechanism was confirmed by immunohistochemical(IHC)and histopathological analysis.RESULTS We identified that Zey dose-dependently decreased cell viability,colony formation and expression of Proliferating Cell Nuclear Antigen(PCNA),and significantly induced K562 cell apoptosis via regulating Bcl-2 family members,decreasing mitochondrial transmembrane potential,and activating caspase-3,caspase-9,and caspase-8(P<0.05 or P<0.01).Further study revealed that Zey significantly inhibited phosphorylation of Jak2 and Src and downregulated their downstream proteins,including stat3,PI3K/AKT/m TOR,and ERK1/2 signaling pathways(P<0.05 or P<0.01).Zey also suppressed tumor growth with low toxicity in mouse xenograft model of K562cells through decreasing expression of Jak2 and Src.CONCLUSION Our data demonstrated that Zey substantially suppressed K562 cells both in vitro and in vivo through Jak2 and Src pathways.These findings suggest the potential of Zey as an effective anticancer agent in CML treatment.
基金Supported by National Natural Science Foundation of China(22264023)Natural Science Foundation of Shaanxi Province(2024JC-YBQN-0150)+2 种基金Yan'an Science and Technology Bureau Project(2023-SFGG-057)Scientific Research Projects of Education Department of Shaanxi Province(22JK0614)PhD Start Fund of Yan'an University(YDBK2022-15)。
文摘Manganese(Mn),an essential trace element in the human body,plays critical roles in many biological processes.Recent studies have discovered that Mn^(2+)may promote or directly activate the cGAS-STING pathway,thereby subsequently initiating the natural immune response and augmenting antitumor therapy.However,the current lack of accurate methods for Mn^(2+)determination in cells significantly limits their mechanism investigation;hence,it is urgent to establish novel tools to detect Mn^(2+)in cells.In this study,the dual-emission carbon dots were initially synthesized via the one-pot hydrothermal method employing L-aspartic acid and p-phenylenediamine as raw materials.In the presence of Mn^(2+),the emission peak centered at 350 nm exhibited significant enhancement,whereas another peak at 610 nm remained stable.Consequently,a ratiometric sensor for Mn^(2+)determination was established using the signal at 350 nm as the responsive signal and the signal at 610 nm as an internal reference.Under the optimal condition,a good linear relationship was achieved between the F350/F610 value and Mn^(2+)concentration ranging from 0.9 to 15μmol/L,with a calculated LOD of 61 nmol/L.Benefiting from the special Mn^(2+)-induced ratiometric approach,this method demonstrates outstanding sensitivity,selectivity,and stability,rendering it applicable for Mn^(2+)determination in complex biological samples,as well as Mn^(2+)imaging in MKN-45 and LO2 cells.
文摘Gastrin and cyclooxygenase-2(COX-2) playimportant roles in the carcinogenesis and progression ofgastric cancer.However,it remains unknown whether the combination of cholecystokinin-2(CCK-2) receptor antagonist plus COX-2 inhibitor exerts synergistic anti-tumor effects on human gastric cancer.Here,we demonstrated that the combination of AG-041R(a CCK-2 receptor antagonist) plus NS-398(a selective COX-2 inhibitor) treatment had synergistic effects on proliferation inhibition,apoptosis induction,down-regulation of Bcl-2 and up-regulation of Bax expression in MKN-45 cells.These results indicate that simultaneous targeting of CCK-2 receptor and COX-2 may inhibit gastric cancer development more effectively than targeting either molecule alone.(C)2008 Elsevier Ireland Ltd.All rights reserved.
文摘Aim The present study developed a CYP3A4-expressed Caco-2 monolayer model at which effects of the efflux-metabolism alliance on the transport and uptake of clausenamide（CLA） enantiomers as CYP3A4 substrates were investigated. The apparent permeability coefficients （Papp） of （ - ） and （ ＋ ）CLA were higher in the ab- sorptive direction than those in the secretory direction with efflux ratios（ER） of 0. 709 ± 0.411 and 0. 867± 0. 250 （ Х10^-6 -1 cm · s ）, respectively. Their bidirectional transports were significantly reduced by （75.6 ± 87.5）% af- ter treatment with verapamil （ a P-glycoprotein inhibitor） that increased the rate of metabolism by CYP3 A4, whereas the CYP3A4 inhibitor ketoconazole treatment markedly enhanced the basolateral to apical flux of （ - ） and （ ＋ ） CLA with ERs being 2. 934 ± 1. 432 and 1. 877 ± 0. 148 （ Х 10^-6 cm/s） respectively. These changes could be blocked by the duel CYP3A4/P-glycoprotein inhibitor cyclosporine A, consequently, Papp values for CLA enanti- omers in both directions were significantly greater than those obtained by using verapamil or ketoconazole, and their ERs were similar to those following （ - ） or （ ＋ ）-isomer treatment alone. Furthermore, the uptake of （ - ）CLA was more than that of （ ＋ ）CLA in the transfected cells. Incubation with ketoeonazole decreased the intracellular concentrations of the two enantiomers. This effect disappeared in the presence of a CYP3A4 inducer dexametha- sone. These results indicated that CYP3A4 could influence P-gp efflux, transport and uptake of CLA enantiomers as CYP3A4 substrates and that a duel inhibition to CYP3A4/ P-glycoprotein could enhance their absorption and bioavailability, which provides new insight into the efflux-metabolism alliance and will benefit the clinical pharma- cology of （？） CLA as a candidate drug for treatment of Alzheimer＇ s disease.
文摘Anatase Ti0_(2) nanosheet-based hierarchical spheres(HSs)with nearly 100%exposed{001}facets were synthesized via a facile solvothermal process.Using these hierarchical spheres as a scattering layer on nanocrystaline TiO_(2)film,hi-layered dye-sensitized solar cells(DSSCs)have been fabricated by electrophoresis deposition method,which well preserved the fragile hierarchical structure.Owing to the superior dye adsorption and light scattering effect of HSs,an overall energy conversion efficiency of 7.38%is achieved,which is 26%higher than that of nanoparticle-based photoanode.
基金supported by National Natural Science Foundation of China(30300253)Chen guang youth technology program of Wuhan(20065004116-25)
文摘Based on published sequences for chicken Bcl-2,three siRNAs（small interfering RNA）were designed,and expression vectors were constructed and transfected into goose granulosa cells cultured in vitro.Bcl-2 protein,apoptosis and proliferation of granulosa cells,48 h after the transf ection,were analyzed by flow cytometry,and progesterone（P）secreted into the culture medium was measured by radioimmunoassay.In addition,apoptosis and Bcl-2 protein level were assessed in untreated granulosa cells from the four largest preovulatory follicles（F<sub>1</sub><sup>F</sup><sub>4</sub>）,the smallest preovulatory follicles（SPF）,small yellow follicles（SYF）and atretic follicles.The highest level of Bcl-2 protein was observed in granulosa cells from SPF,and levels in cells from healthy follicles were significantly higher than those of atretic follicles（P【0.05）.Bcl-2 protein levels in cells subjected to RNAi were significantly lower than those of controls（P【0.05）,while apoptosis indices（AI）,proliferation indices（PI）and P secretion in the RNAi treatments were higher than those of controls（P【0.05）.
基金The project supported by the National Natural Science Foundation of China(81072542)Natural Science Foundation of Jiangsu Province(BK2011077)the Research Fund for the Doctoral Program of Higher Education(20123237110010)
文摘OBJECTIVE Liguzinediol is a derivative of the natural active ingredient ligustrazine,and we found that liguzinediol has significant positive inotropic effects,which are stronger than that of TMP.Besides,it does not lead to arrhythmia,hypotension and other side effects.This study aims to investigate the anti-apoptotic effects of liguzinediolon H9C2 cells.METHODS Apoptotic H9C2 cells induced by DOX were observed by electron microscope and FCM analysis.The protein expressions of Bax,Bcl-2,caspases 3 and NF-κB were detected by Western blotting.RESULTS Apoptotic H9C2 cells induced by DOX were observed,but without apoptotic bodies in liguzinediol group.Declined peak of H9C2 cell apoptosis was seen in liguzinediol group by FCM analysis.And downregulation of Bax,caspases 3,NF-κB and upregulation of Bcl-2 were found by Western blotting.CONCLUSION Liguzinediol protected cardiomyocytes against apoptosis through downregulation of Bax and caspases 3 and upregulation of Bcl-2.Liguzinediol can inhibited cardiomyocyte apoptosis through the NF-κB signal pathway.
文摘OBJECTIVE Hepatocel ular carcinoma(HCC)is the most common cause of cancer-related mortality,with high incidence rates,robust metastatic propensity and acquired resistance to therapy.Metformin,an extensively prescribed and well-tolerated first-linetherapeutic drug for type 2 diabetes mellitus,has recently been identified as a potential and attractive anticancer adjuvant drug combined with chemotherapeutics to improve treatment efficacy and lower doses.Curcumin,a botanical extracts,has been shown antitumorigenic properties.This study aims to investigate the combinational effect of metformin and curcumin on inbibition of tumor growth and metastasis in Hep G2 cells and the possible underlying mechanisms.METHODS The cell proliferation was determined by MTT,CCK-8 and colony formation assay.The protein expression was detected by Western blotting.Activity of MMP-2 and MMP-9 was estimated by gelatin zymography.Flow cytometry analysis was used to evaluate the influence of metformin and curcumin on cell cycle arrest and apoptosis,and morphology observation of apoptosis was detected by Hoechst33342.Scratch and transwell assay was performed to detect the cell migration and invasion.The suppression of this combination therapy oncapillary tube formation was detected by tube formation assay.RESULTS Combination of metformin and curcumin induced stronger inhibition on Hep G2 cells proliferation than monotherapywhich related to induction of cell cycle arrest in G2/M phase and apoptosis through regulation of the protein expression of cyclin B and Bcl-2/Bax.Moreover,the co-treatment of metformin with curcumin exerted an enhanced inhibitory effect on Hep G2 cell metastasis and synergistically inhibited the tube formation of HUVEC cells.The suppression of PI3K/AKT/m TOR pathway and inhibition the protein expression of STAT3,MMP9,MMP2 and VEGF might involve in this synergistic effects of combination treatment.CONCLUSION Combination of metformin and curcumin inhibited Hep G2 cells proliferationmore effectively than monotherapy and synergistically induced a greater inhibition on migration and invasion of Hep G2 cells.
基金The project supported by National Natural Science Foundation of China(81473383,81573645)
文摘OBJECTIVE To investigate the effects of ex-tract of Ramulus Cinnamom(RC)against LPS-induced inflammation in microglia.METHODS Activated microglia releases various pro-inflammatory cytokines to induce neuroinflammation in stroke.Lipopolysaccaride(LPS)is an endotoxin from the outer membrane of Gram-negative bacteria that activates microglia.MTT assay was used to observe the cell viability.The content of NO in supernatant was measured by Griess reagent.The levels of IL-1β,IL-6 and TNF-αin supernatant were detected by ELISA kits.The intracellular COX-2,TLR4,and My D88expression was assayed by Western blotting.RESULTS RC extract 30 and 100μg·m L-1significantly decreased the production of related inflammatory factors such as NO(P<0.05,P<0.01),IL-1β(P<0.01,P<0.01),IL-6(P<0.05,P<0.01)and TNF-α(P<0.01,P<0.01).Furthermore,RC extract significantly inhibited the COX-2,TLR4,and My D88 expression induced by LPS in BV2cells.CONCLUSION RC extract may have therapeutic potential for the improvement of neuroinflammation,and the mechanism may be involved in down-regulation of TLR4/My D88 inflammation pathway.
文摘OBJECTIVE Aconitine(ACO)as the main active component in Aconitum carmichaelii debeaux(family Ranunlaceae),has highly toxicity in heart and the mechanisms are not clear yet.Paeoniflorin(PF),the main chemical ingredient in Herbaceous peony,can protect heart hurt by antioxidant,vasodilator effect and other effects.In this study,we focused on investigating the mechanism of PF reducing the cardiotoxicity of ACO.METHODS We chose H9c2 cells as experimental subject.MTT,Western blotting and real-time PCR were used to measure cell proliferation,apoptosis,ion channels and oxidative stress.RESULTS Cell proliferation in ACO+PF group was significantly increased compared with ACO group;the ratio with Bcl-2 and Bax and the level of p53 were upregulated by PF,while the level of caspase-3 was lightly reduced.The expression of SCN5A mRNA significantly was increased in ACO+PF group,while the expres⁃sion of RyR2 and Cx43 mRNA was dropped.Compared with ACO group,extracellular LDH and intracellular MDA were highly decreased,while intracellular SOD was regulated.CONCLUSION Cardiotoxicity of ACO in H9c2 cells was signifi⁃cantly decreased by PF.
基金The project supported by National Natural Science Foundation of China(81260664,81160538)
文摘OBJECTIVE To investigate the effect of P2X7receptor(P2X7r)inhibition,using a specific inhibitor(A438079)to prevent the development of liver fibrosis on human hepatic stellate cells,LX-2.METHODS The supernatant from lipopolysaccharide(LPS)-stimulated RAW264.7 mouse macrophages was supplemented to LX-2 cells for 24 h.LX-2cells were primed with LPS for 4h and subsequently stimulated for 30 min with 3mmol·L-1 of adenosine 5′-triphosphate(ATP).A438079(10μmol·L-1)was supplemented to LX-2 cells 10 min prior to ATP.RESULTS Directly treated with LPS on LX-2 cells,mRNA expressions of IL-1β,IL-18 and IL-6 were increased,as well as P2X7 r.And caspase-1,ASC and NLRP3 mRNA expressions were increased with LPS stimulation.LPS stimulation also increasedα-SMA and collagenⅠ mRNA expressions.Interestingly treatment of LX-2cells with mediums from LPS-primed RAW264.7mouse macrophages exhibited greater increase of mRNA expressions of above genes than those in LX-2directly treated with LPS.Pretreatment of directly or indirectly LPS-stimulated LX-2 cells with A438079 both suppressed IL-1βmRNA expression.In addition treatment of LPS-primed LX-2 cells with 3mmol·L-1 ATP induced the significant increase of IL-1β,IL-6,caspase-1,pannexin-1,α-SMA and collagenⅠ mRNA expression,the increasing ofα-SMA protein expression and cleavage of IL-1β.These events were significantly suppressed by pretreatment with P2X7 rantagonist A438079.P2X7 rblockade also significantly reduced the protein expression ofα-SMA.CONCLUSION Our results suggest that the involvement of the P2X7r-NLRP3 inflammasome pathway in the secretion of IL-1βfrom extracellular ATP/LPS-stimulated human hepatic stellate cells.This study demonstrated that repression of the P2X7 rrepresents a novel potential therapeutic approach to control liver fibrosis.
文摘OBJECTIVE A novel mast cell-specific G-protein-coupled receptor(GPCR),known as mas-related GPCR-B2(MRGPRB2),plays important roles in the immune response.The opening of ion channels mediated by MRGPRB2 activation remains unclear.METHODS AND RE⁃SULTS Calcium influx induced by activation of MRGPRB2 receptor in mouse peritoneal mast cells was related to the concentrations of calcium ions in the extracellular solution.Similarly,the volt⁃age-dependent current generated by MRGPRB2 activation was also correlated with the extracellu⁃lar calcium concentration.In addition,the in⁃creased of calcium influx or voltage-dependent current caused by activation of MrgprB2 could be blocked by U73122(PLC blocker)or 2-APB(IP3-ORAI1 blocker).Meanwhile,calcium-activated chlorine channel(TMEM16A)was involved in the generation of voltage-dependent currents in⁃duced by MRGPRB2 activation in mouse perito⁃neal mast cells.Furthermore,the degranulation of mouse peritoneal mast cells mediated by MRGPRB2 receptor could also be inhibited by U73122 or 2-APB.CONCLUSION PLC-IP3-ORAI1-TMEM16A signaling pathway was involved in MRGPRB2-mediated mast cell activation.
文摘P2X7 receptor （P2X7r） is important in inflammation and fibrosis. The aim of the present study was to investigate the effect of P2X7r inhibition, using a specific inhibitor （A438079） to prevent the development of liver fibrosis on human hepatic stellate cells, LX-2. The supernatant from lipopolysaccharide （LPS）-stimulated RAW 264.7 mouse macrophages was supplemented to LX-2 cells for 24 h. LX-2 cells were primed with LPS for 4 h and subsequently stimulated for 30 rain with 3 mmol · L^-1 of adenosine 5＇-triphosphate （ATP）. A438079 （ 10 μmol · L^-1） was supplemented to LX-2 cells 10 rain prior to ATP. Directly treated with LPS on LX-2 cells, mRNA ex- pressions of IL-1β, IL-18 and IL-6 were increased, as well as P2X7r. And caspase-1, ASC and NLRP3 mRNA ex- pressions were increased with LPS stimulation. LPS stimulation also increased oL-SMA and collagen I mRNA expres- sions. Interestingly treatment of LX-2 cells with mediums from LPS-primed RAW 264.7 mouse macrophages exhibi- ted greater increase of mRNA expressions of above genes than those in LX-2 directly treated with LPS. Pretreatment of directly or indirectly LPS-stimulated LX-2 cells with A438079 both suppressed IL-1β mRNA expression. In addi- tion treatment of LPS-primed LX-2 cells with 3 mM ATP induced the significant increase of IL-1β, IL-6, caspase- 1, pannexin-1, α-SMA and collagen I mRNA expression, the increasing of oL-SMA protein expression and cleavage of IL-1β. These events were significantly suppressed by pretreatment with P2X7r antagonist A438079. P2XTr blockade also significantly reduced the protein expression of oL-SMA. Our results suggest that the involvement of the P2X7r-NLRP3 inflammasome pathway in the secretion of IL-1β from extracellular ATP/LPS-stimulated human he- patic stellate cells. This study demonstrated that repression of the P2XTr represents a novel potential therapeutic ap- proach to control liver fibrosis.
基金Space Application System of China Manned Space Program。
文摘Significant progress has recently been made in enhancing the power conversion efficiency(PCE)of perovskite solar cells(PSCs).The electron transport layer(ETL),as an essential component of PSCs,significantly influences the performance of devices.Traditional spin-coating method for preparing the ETL fails to fully cover the cusp of FTO transparent conductive glass substrate,leading to direct contact between perovskite film and FTO substrate,which induces charge recombination and reduces the performance of PSCs.To address this issue,an in-situ growth method was proposed to prepare conformal SnO_(2) films on FTO glass substrates in this study.The resulting SnO_(2) films are not only dense and uniform,fully covering the cusp of the FTO glass substrates and reducing the contact area between the FTO substrates and the perovskite films,but also facilitating the formation of perovskite films with large grain sizes.Moreover,the conformal SnO_(2) films can improve the charge extraction at the SnO_(2)/perovskite interface,reduce the trap density and trap-assisted recombination in PSCs,and thus enhance the PCE of PSCs.Through comparative experiments,it is found that the PSCs with in-situ grown SnO_(2) films show an improved PCE of 21.97%,which significantly increased compared to that with spin-coated SnO_(2) films(20.93%).All above data demonstrate that the as-prepared SnO_(2) film can serve as an ideal ETL.It is worth mentioning that this method avoids the use of corrosive hydrochloric acid and toxic thioglycolic acid,and it can also be extended to ITO flexible transparent conductive substrates in the future.
基金Supported by the National Natural Science Foundation of China(31201911 31372438)
文摘Porcine epidemic diarrhea(PED)is caused by porcine epidemic diarrhea virus(PEDV),and is characterized by vomiting,diarrhea and dehydration of suckling pigs from 80% to 100% morbidity and 50% to 90% mortality,and resulted in tremendous economic losses to swine industry.The PEDV mainly infects small intestine of pigs,resulting in vacuolar degeneration and necrosis of mucosal epithelium.The IPEC-J2 is a pig intestine epithelial cell line,which is similar to the intestinal environment of piglets,can be used to isolate and identify the PEDV field isolates.In this study,it appeared the PEDV typical postmortem changes and histopathological lesion of degeneration and destruction of small intestine in infected piglets,and IHC identified that the PEDV distributed in the mucosa and submucosa of small intestine mostly.Furthermore,the PEDV HLJ strain was successfully isolated and characterized in the IPEC-J2 cells,and indicated that the IPEC-J2 cell line was sensitive to isolate and adapt the PEDV field strain,and could be utilized to multiply the PEDV rapidly.The S gene analysis indicated that the PEDV HLJ strain was the prevailed virus,belonged to Group 1 with attenuated virulent DR13,SC1402 and J-S2/2015 strains isolated in South Korea and China from 2014 to 2015.This study had important theoretical and practical significances on analyzing genetic variation of the PEDV,understanding the pathogenic characteristics of the virus and developing new vaccines for the PED.
文摘MoS_(2)/CuS composite catalysts were successfully synthesized using a one-step hydrothermal method with sodium molybdate dihydrate,thiourea,oxalic acid,and copper nitrate trihydrate as raw materials.The hydrogen pro-duction performance of MoS_(2)/CuS prepared with different molar ratios of Mo to Cu precursors(n_(Mo)∶n_(Cu))as cathodic catalysts was investigated in the two-chamber microbial electrolytic cell(MEC).X-ray diffraction(XRD),X-ray pho-toelectron spectroscopy(XPS),scanning electron microscopy(SEM),transmission electron microscope(TEM),linear scanning voltammetry(LSV),electrochemical impedance analysis(EIS),and cyclic voltammetry(CV)were used to characterize the synthesized catalysts for testing and analyzing the hydrogen-producing performance.The results showed that the hydrogen evolution performance of MoS_(2)/CuS-20%(nMo∶nCu=5∶1)was better than that of platinum(Pt)mesh,and the hydrogen production rate of MoS_(2)/CuS-20%as a cathode in MEC was(0.2031±0.0237)m^(3)_(H_(2))·m^(-3)·d^(-1) for 72 h at an applied voltage of 0.8 V,which was slightly higher than that of Pt mesh of(0.1886±0.0134)m^(3)_(H_(2))·m^(-3)·d^(-1).The addition of a certain amount of CuS not only regulates the electron transfer ability of MoS_(2) but also increases the density of active sites.
