Objective This study aimed to establish a neural cell injury model in vitro by stimulating PC12 cells with lipopolysaccharide(LPS)and to examine the effects of astragaloside IV on key targets using high-throughput seq...Objective This study aimed to establish a neural cell injury model in vitro by stimulating PC12 cells with lipopolysaccharide(LPS)and to examine the effects of astragaloside IV on key targets using high-throughput sequence technology and bioinformatics analyses.Methods PC12 cells in the logarithmic growth phase were treated with LPS at final concentrations of 0.25,0.5,0.75,1,and 1.25 mg/mL for 24 h.Cell morphology was evaluated,and cell survival rates were calculated.A neurocyte inflammatory model was established with LPS treatment,which reached a 50%cell survival rate.PC12 cells were treated with 0.01,0.1,1,10,or 100µmol/L astragaloside IV for 24 h.The concentration of astragaloside IV that did not affect the cell survival rate was selected as the treatment group for subsequent experiments.NOS activity was detected by colorimetry;the expression levels of ERCC2,XRCC4,XRCC2,TNF-α,IL-1β,TLR4,NOS and COX-2 mRNA and protein were detected by RT-qPCR and Western blotting.The differentially expressed genes(DEGs)between the groups were screened using a second-generation sequence(fold change>2,P<0.05)with the following KEGG enrichment analysis,RT-qPCR and Western blotting were used to detect the mRNA and protein expression of DEGs related to the IL-17 pathway in different groups of PC12 cells.Results The viability of PC12 cells was not altered by treatment with 0.01,0.1,or 1µmol/L astragaloside IV for 24 h(P>0.05).However,after treatment with 0.5,0.75,1,or 1.25 mg/mL LPS for 24 h,the viability steadily decreased(P<0.01).The mRNA and protein expression levels of ERCC2,XRCC4,XRCC2,TNF-α,IL-1β,TLR4,NOS,and COX-2 were significantly increased after PC12 cells were treated with 1 mg/mL LPS for 24 h(P<0.01);however,these changes were reversed when PC12 cells were pretreated with 0.01,0.1,or 1µmol/L astragaloside IV in PC12 cells and then treated with 1 mg/mL LPS for 24 h(P<0.05).Second-generation sequencing revealed that 1026 genes were upregulated,while 1287 genes were downregulated.The DEGs were associated with autophagy,TNF-α,interleukin-17,MAPK,P53,Toll-like receptor,and NOD-like receptor signaling pathways.Furthermore,PC12 cells treated with a 1 mg/mL LPS for 24 h exhibited increased mRNA and protein expression of CCL2,CCL11,CCL7,MMP3,and MMP10,which are associated with the IL-17 pathway.RT-qPCR and Western blotting analyses confirmed that the DEGs listed above corresponded to the sequence assay results.Conclusion LPS can damage PC12 cells and cause inflammatory reactions in nerve cells and DNA damage.astragaloside IV plays an anti-inflammatory and DNA damage protective role and inhibits the IL-17 signaling pathway to exert a neuroprotective effect in vitro.展开更多
Non-linear numerical modeling, widely used in research and development to understand many complex processes such as forming or machining, does not guarantee the success of a study to be performed. Indeed, the numerica...Non-linear numerical modeling, widely used in research and development to understand many complex processes such as forming or machining, does not guarantee the success of a study to be performed. Indeed, the numerical simulation uses finite element codes where the models already integrated are not based on shapes adjustable to any type of study. In this study, a new form of non-linear constitutive flow law based on the Modified Zerilli-Armstrong model, which can answer the above problem, has been developed to apply it to the numerical simulation of two different tests (a quasi-static compression test, the necking of a circular bar). This flow law is based on the modified Zerilli-Armstrong model, which, together with the new modified Johnson-Cook model, has been compared to appreciate the relevance of the proposal. For that, an implementation of this new law via the VUHARD subroutine into the Abaqus/Explicit finite element code was made to model the two tests. The comparison of the results obtained (from identification) by our proposed law with those obtained using the NMJC shows that this new law better approaches the experiments than the other one. This is also shown through the numerical results using the Abaqus software. It can be said that this way of formulating a flow law allows highlighting the great performance of the proposed approach. Although this law has been only used for quasi-static tests, we can say that it can also be used in dynamic tests.展开更多
Probiotic formulations containing lecithin phospholipids have been shown to extend their viability.However,the protective effect of lecithin and the properties of the mixtures of lecithin and probiotic cells in aqueou...Probiotic formulations containing lecithin phospholipids have been shown to extend their viability.However,the protective effect of lecithin and the properties of the mixtures of lecithin and probiotic cells in aqueous media have not been fully documented.The aim of this study was to investigate the interactions between Bifidobacterium animalis ssp.lactis(Bifido)probiotics and lecithin.The lecithin-Bifido interactions were dependent on lecithin concentration.FTIR and Raman spectroscopy suggested that Bifido cells interacted with lecithin molecules,mainly through hydrophobic interactions.At the lecithin concentration of 0.1%.w/v,Bifido lost only about 17%of their viability over 56 days,while free Bifido probiotics(without lecithin)lost about 66%of their viability in the same time period.Furthermore,flow cytometer side scatter plots also showed that Bifido-lecithin samples at 0.1.%w/v lecithin,showed the lowest side scatter,which is indicative of lower cellular stress and a more stable environment to preserve Bifido cells viability over time.展开更多
The authors regret that the incorrect Fig.7/page 6,and Fig.8/page 7 were published.The correct figures with appropriate naming and scale are provided in this corrigendum.This correction has not changed the description...The authors regret that the incorrect Fig.7/page 6,and Fig.8/page 7 were published.The correct figures with appropriate naming and scale are provided in this corrigendum.This correction has not changed the description,or the original conclusions of the article.展开更多
基金supported by grants from Open Project of Gansu Traditional Chinese Medicine Research Center(No.zyzx-2020-10)Gansu Province Youth Science and Technology Foundation Program(No.21JR7RA652)+1 种基金Gansu Province Higher Education Research(No.2018A-049)Gansu Province Higher Education Research(No.2021B-163).
文摘Objective This study aimed to establish a neural cell injury model in vitro by stimulating PC12 cells with lipopolysaccharide(LPS)and to examine the effects of astragaloside IV on key targets using high-throughput sequence technology and bioinformatics analyses.Methods PC12 cells in the logarithmic growth phase were treated with LPS at final concentrations of 0.25,0.5,0.75,1,and 1.25 mg/mL for 24 h.Cell morphology was evaluated,and cell survival rates were calculated.A neurocyte inflammatory model was established with LPS treatment,which reached a 50%cell survival rate.PC12 cells were treated with 0.01,0.1,1,10,or 100µmol/L astragaloside IV for 24 h.The concentration of astragaloside IV that did not affect the cell survival rate was selected as the treatment group for subsequent experiments.NOS activity was detected by colorimetry;the expression levels of ERCC2,XRCC4,XRCC2,TNF-α,IL-1β,TLR4,NOS and COX-2 mRNA and protein were detected by RT-qPCR and Western blotting.The differentially expressed genes(DEGs)between the groups were screened using a second-generation sequence(fold change>2,P<0.05)with the following KEGG enrichment analysis,RT-qPCR and Western blotting were used to detect the mRNA and protein expression of DEGs related to the IL-17 pathway in different groups of PC12 cells.Results The viability of PC12 cells was not altered by treatment with 0.01,0.1,or 1µmol/L astragaloside IV for 24 h(P>0.05).However,after treatment with 0.5,0.75,1,or 1.25 mg/mL LPS for 24 h,the viability steadily decreased(P<0.01).The mRNA and protein expression levels of ERCC2,XRCC4,XRCC2,TNF-α,IL-1β,TLR4,NOS,and COX-2 were significantly increased after PC12 cells were treated with 1 mg/mL LPS for 24 h(P<0.01);however,these changes were reversed when PC12 cells were pretreated with 0.01,0.1,or 1µmol/L astragaloside IV in PC12 cells and then treated with 1 mg/mL LPS for 24 h(P<0.05).Second-generation sequencing revealed that 1026 genes were upregulated,while 1287 genes were downregulated.The DEGs were associated with autophagy,TNF-α,interleukin-17,MAPK,P53,Toll-like receptor,and NOD-like receptor signaling pathways.Furthermore,PC12 cells treated with a 1 mg/mL LPS for 24 h exhibited increased mRNA and protein expression of CCL2,CCL11,CCL7,MMP3,and MMP10,which are associated with the IL-17 pathway.RT-qPCR and Western blotting analyses confirmed that the DEGs listed above corresponded to the sequence assay results.Conclusion LPS can damage PC12 cells and cause inflammatory reactions in nerve cells and DNA damage.astragaloside IV plays an anti-inflammatory and DNA damage protective role and inhibits the IL-17 signaling pathway to exert a neuroprotective effect in vitro.
文摘Non-linear numerical modeling, widely used in research and development to understand many complex processes such as forming or machining, does not guarantee the success of a study to be performed. Indeed, the numerical simulation uses finite element codes where the models already integrated are not based on shapes adjustable to any type of study. In this study, a new form of non-linear constitutive flow law based on the Modified Zerilli-Armstrong model, which can answer the above problem, has been developed to apply it to the numerical simulation of two different tests (a quasi-static compression test, the necking of a circular bar). This flow law is based on the modified Zerilli-Armstrong model, which, together with the new modified Johnson-Cook model, has been compared to appreciate the relevance of the proposal. For that, an implementation of this new law via the VUHARD subroutine into the Abaqus/Explicit finite element code was made to model the two tests. The comparison of the results obtained (from identification) by our proposed law with those obtained using the NMJC shows that this new law better approaches the experiments than the other one. This is also shown through the numerical results using the Abaqus software. It can be said that this way of formulating a flow law allows highlighting the great performance of the proposed approach. Although this law has been only used for quasi-static tests, we can say that it can also be used in dynamic tests.
基金support provided by the Innovation Fund Denmark(PROBIO project-7076-00053B).
文摘Probiotic formulations containing lecithin phospholipids have been shown to extend their viability.However,the protective effect of lecithin and the properties of the mixtures of lecithin and probiotic cells in aqueous media have not been fully documented.The aim of this study was to investigate the interactions between Bifidobacterium animalis ssp.lactis(Bifido)probiotics and lecithin.The lecithin-Bifido interactions were dependent on lecithin concentration.FTIR and Raman spectroscopy suggested that Bifido cells interacted with lecithin molecules,mainly through hydrophobic interactions.At the lecithin concentration of 0.1%.w/v,Bifido lost only about 17%of their viability over 56 days,while free Bifido probiotics(without lecithin)lost about 66%of their viability in the same time period.Furthermore,flow cytometer side scatter plots also showed that Bifido-lecithin samples at 0.1.%w/v lecithin,showed the lowest side scatter,which is indicative of lower cellular stress and a more stable environment to preserve Bifido cells viability over time.
文摘The authors regret that the incorrect Fig.7/page 6,and Fig.8/page 7 were published.The correct figures with appropriate naming and scale are provided in this corrigendum.This correction has not changed the description,or the original conclusions of the article.
