Objective To investigate the effects of microRNA-18a(miR-18a) on migration and invasion of hepatocellular carcinoma(HCC) cells,and its possible mechanism associated with Dicer l. Methods HepG 2 and HepG 2.2.15 cells w...Objective To investigate the effects of microRNA-18a(miR-18a) on migration and invasion of hepatocellular carcinoma(HCC) cells,and its possible mechanism associated with Dicer l. Methods HepG 2 and HepG 2.2.15 cells were transfected with miR-18 a inhibitor using Lipofectamine. Cell invasion was evaluated by transwell invasion assay,and cell migration was detected by transwell migration and wound-healing assays. Moreover,luciferase reporter assay was used to identify whether Dicer expression was regulated by miR-18 a. Real-time RT-PCR and western blot were performed to analyze Dicer 1 expression. In addition,a functional restoration assay was performed to investigate whether miR-18 a promotes HCC cell migration and invasion by directly targeting Dicer 1. Results miR-18 a inhibitor can suppress the migration and invasion of HCC cells. Furthermore,suppression of Dicer l expression by small interfering RNA essentially abolished the inhibition of cell migration and invasion induced by miR-18 a inhibitor,restorating these activities to levels similar to the parental HCC cells. Interestingly,suppression of miR-18 a in HCC cells resulted in enhanced expression of Dicer l. In addition,the results of a luciferase assay demonstrated targeted regulation of Dicer l by miR-18 a. Conclusion Our findings suggest that miR-18 a promotes migration and invasion of HCC cells by inhibiting Dicer l expression.展开更多
Objective Huangqi Decoction(HQD),a classical traditional Chinese medicine formula,has been used as a valid treatment for alleviating liver fibrosis;however,the underlying molecular mechanism is still unknown.Although ...Objective Huangqi Decoction(HQD),a classical traditional Chinese medicine formula,has been used as a valid treatment for alleviating liver fibrosis;however,the underlying molecular mechanism is still unknown.Although our previous studies showed that microRNA-663a(miR-663a)suppresses the proliferation and activation of hepatic stellate cells(HSCs)and the transforming growth factor-β/small mothers against decapentaplegic(TGF-β/Smad)pathway,whether long noncoding RNAs(lncRNAs)are involved in HSC activation via the miR-663a/TGF-β/Smad signaling pathway has not yet reported.The present study aimed to investigate the roles of lncRNA lnc-C18orf26-1 in the activation of HSCs and the mechanism by which HQD inhibits hepatic fibrosis.Methods The expression levels of lnc-C18orf26-1,miR-663a and related genes were measured by quantitative reverse transcription-polymerase chain reaction.HSCs were transfected with the miR-663a mimic or inhibitor and lnc-C18orf26-1 small interfering RNAs.The water-soluble tetrazolium salt-1 assay was used to assess the proliferation rate of HSCs.Changes in lncRNA expression were evaluated in miR-663a-overexpressing HSCs by using microarray to identify miR-663a-regulated lncRNAs.RNA hybrid was used to predict the potential miR-663a binding sites on lncRNAs.Luciferase reporter assays further confirmed the interaction between miR-663a and the lncRNA.The expression levels of collagen α-2(I)chain(COL1A2),α-smooth muscle actin(α-SMA)and TGF-β/Smad signaling pathway-related proteins were determined using Western blotting.Results Lnc-C18orf26-1 was upregulated in TGF-β1-activated HSCs and competitively bound to miR-663a.Knockdown of lnc-C18orf26-1 inhibited HSC proliferation and activation,downregulated TGF-β1-stimulatedα-SMA and COL1A2 expression,and inhibited the TGF-β1/Smad signaling pathway.HQD suppressed the proliferation and activation of HSCs.HQD increased miR-663a expression and decreased lnc-C18orf26-1 expression in HSCs.Further studies showed that HQD inhibited the expression of COL1A2,α-SMA,TGF-β1,TGF-βtype I receptor(TGF-βRI)and phosphorylated Smad2(p-Smad2)in HSCs,and these effects were reversed by miR-663a inhibitor treatment.Conclusion Our study identified lnc-C18orf26-1 and miR-663a as promising therapeutic targets for hepatic fibrosis.HQD inhibits HSC proliferation and activation at least partially by regulating the lnc-C18orf26-1/miR-663a/TGF-β1/TGF-βRI/p-Smad2 axis.展开更多
The sorption study of La(Ⅲ) was carried out on poly[dibenzo-18-crown-6] and L-valine medium. The quantitative adsorption of La(Ⅲ) was found at 1×10-2 to 1×10-5 mol/L L-valine. The various eluting agents we...The sorption study of La(Ⅲ) was carried out on poly[dibenzo-18-crown-6] and L-valine medium. The quantitative adsorption of La(Ⅲ) was found at 1×10-2 to 1×10-5 mol/L L-valine. The various eluting agents were found efficient eluents for La(Ⅲ). The capacity of crown polymer for La(Ⅲ) was found to be 0.43 ±0.01 mmol/g. The tolerance limit of various cations and anions for La(Ⅲ) was determined. La(Ⅲ) was quantitatively separated from other metal ions in binary as well as multicomponent mixtures. The study was extended to sequential separation of La(Ⅲ), U(VI) and Th(IV). The good separation yields were obtained and had good reproducibility (±2%). The method incorporated the determination of La(Ⅲ) in real sample. The method was simple, rapid and selective.展开更多
After the strong 2015/16 El Nino event,cold conditions prevailed in the tropical Pacific with the second-year cooling of the 2017/18 La Ni?a event.Many coupled models failed to predict the cold SST anomalies(SSTAs)in ...After the strong 2015/16 El Nino event,cold conditions prevailed in the tropical Pacific with the second-year cooling of the 2017/18 La Ni?a event.Many coupled models failed to predict the cold SST anomalies(SSTAs)in 2017.By using the ERA5 and GODAS(Global Ocean Data Assimilation System)products,atmospheric and oceanic factors were examined that could have been responsible for the second-year cooling,including surface wind and the subsurface thermal state.A time sequence is described to demonstrate how the cold SSTAs were produced in the central-eastern equatorial Pacific in late 2017.Since July 2017,easterly anomalies strengthened in the central Pacific;in the meantime,wind stress divergence anomalies emerged in the far eastern region,which strengthened during the following months and propagated westward,contributing to the development of the second-year cooling in 2017.At the subsurface,weak negative temperature anomalies were accompanied by upwelling in the eastern equatorial Pacific,which provided the cold water source for the sea surface.Thereafter,both the cold anomalies and upwelling were enhanced and extended westward in the centraleastern equatorial Pacific.These changes were associated with the seasonally weakened EUC(the Equatorial Undercurrent)and strengthened SEC(the South Equatorial Current),which favored more cold waters being accumulated in the central-equatorial Pacific.Then,the subsurface cold waters stretched upward with the convergence of the horizontal currents and eventually outcropped to the surface.The subsurface-induced SSTAs acted to induce local coupled air–sea interactions,which generated atmospheric–oceanic anomalies developing and evolving into the second-year cooling in the fall of 2017.展开更多
Objective:Cytotoxic T lymphocytes(CTLs)play a crucial role in the therapeutic approach to hepatocellular carcinoma(HCC).Recent research has indicated that junctional adhesion molecule-like protein(JAML)enhances the an...Objective:Cytotoxic T lymphocytes(CTLs)play a crucial role in the therapeutic approach to hepatocellular carcinoma(HCC).Recent research has indicated that junctional adhesion molecule-like protein(JAML)enhances the antitumor activity of CD8+T cells.Our study investigates the role of JAML+CD8+T cells in HCC.Methods:We utilized time-of-flight mass cytometry and an orthotopic mouse model of HCC to examine histone modifications in tumor-infiltrating immune cells undergoing immunotherapy.Flow cytometry was used to assess CD4+T cells differentiation and JAML expression in CD8+T cells infiltrating HCC.Correlation analysis revealed a strong positive correlation between lactate dehydrogenase A+(LDHA+)CD4+T cells and JAML+CD8+T cells.Subsequently,we evaluated the therapeutic effects of an agonistic anti-JAML antibody,both alone and combined with immunotherapy.Finally,RNA sequencing was conducted to identify potential regulatory mechanisms.Results:Immunotherapy significantly increased the percentage of CD8+T cells infiltrating HCC and induced histone modifications,such as H3K18 lactylation(H3K18la)in CD4+T cells.Flow cytometry analysis revealed that lactate promotes the differentiation of CD4+T cells into Th1 cells.LDHA,an enzyme that converts pyruvate to lactate,plays a key role in this process.Correlation analysis revealed a strong positive relationship between LDHA+CD4+T cells and JAML+CD8+T cells in patients who responded to immunotherapy.Moreover,high JAML expression in CD8+T cells was associated with a more favorable prognosis.In vivo experiments demonstrated that agonistic anti-JAML antibody therapy reduced tumor volume and significantly prolonged the survival of tumor-bearing mice,independent of the effects of anti-programmed cell death protein ligand-1 antibody(αPD-L1)-mediated immunotherapy.Pathway enrichment analysis further revealed that JAML enhances CTL responses through the oxidative phosphorylation pathway.Conclusions:Activation of JAML enhances CTL responses in HCC treatment,independent ofαPD-L1-mediated immunotherapy,providing a promising strategy for advanced HCC.展开更多
基金Supported by the National Natural Science Foundation of China(81472485)the Project of Six Talent Peaks of Jiangsu Province(WSW-014)Project of Hospital Management Center of Wuxi(YGZXQ1311,YGZ1106,and YGZ1007)
文摘Objective To investigate the effects of microRNA-18a(miR-18a) on migration and invasion of hepatocellular carcinoma(HCC) cells,and its possible mechanism associated with Dicer l. Methods HepG 2 and HepG 2.2.15 cells were transfected with miR-18 a inhibitor using Lipofectamine. Cell invasion was evaluated by transwell invasion assay,and cell migration was detected by transwell migration and wound-healing assays. Moreover,luciferase reporter assay was used to identify whether Dicer expression was regulated by miR-18 a. Real-time RT-PCR and western blot were performed to analyze Dicer 1 expression. In addition,a functional restoration assay was performed to investigate whether miR-18 a promotes HCC cell migration and invasion by directly targeting Dicer 1. Results miR-18 a inhibitor can suppress the migration and invasion of HCC cells. Furthermore,suppression of Dicer l expression by small interfering RNA essentially abolished the inhibition of cell migration and invasion induced by miR-18 a inhibitor,restorating these activities to levels similar to the parental HCC cells. Interestingly,suppression of miR-18 a in HCC cells resulted in enhanced expression of Dicer l. In addition,the results of a luciferase assay demonstrated targeted regulation of Dicer l by miR-18 a. Conclusion Our findings suggest that miR-18 a promotes migration and invasion of HCC cells by inhibiting Dicer l expression.
基金supported by grants from the National Natural Science Foundation of China (No.82074101 and No.81773979)Shanghai Municipal Health Commission (No.202040486)
文摘Objective Huangqi Decoction(HQD),a classical traditional Chinese medicine formula,has been used as a valid treatment for alleviating liver fibrosis;however,the underlying molecular mechanism is still unknown.Although our previous studies showed that microRNA-663a(miR-663a)suppresses the proliferation and activation of hepatic stellate cells(HSCs)and the transforming growth factor-β/small mothers against decapentaplegic(TGF-β/Smad)pathway,whether long noncoding RNAs(lncRNAs)are involved in HSC activation via the miR-663a/TGF-β/Smad signaling pathway has not yet reported.The present study aimed to investigate the roles of lncRNA lnc-C18orf26-1 in the activation of HSCs and the mechanism by which HQD inhibits hepatic fibrosis.Methods The expression levels of lnc-C18orf26-1,miR-663a and related genes were measured by quantitative reverse transcription-polymerase chain reaction.HSCs were transfected with the miR-663a mimic or inhibitor and lnc-C18orf26-1 small interfering RNAs.The water-soluble tetrazolium salt-1 assay was used to assess the proliferation rate of HSCs.Changes in lncRNA expression were evaluated in miR-663a-overexpressing HSCs by using microarray to identify miR-663a-regulated lncRNAs.RNA hybrid was used to predict the potential miR-663a binding sites on lncRNAs.Luciferase reporter assays further confirmed the interaction between miR-663a and the lncRNA.The expression levels of collagen α-2(I)chain(COL1A2),α-smooth muscle actin(α-SMA)and TGF-β/Smad signaling pathway-related proteins were determined using Western blotting.Results Lnc-C18orf26-1 was upregulated in TGF-β1-activated HSCs and competitively bound to miR-663a.Knockdown of lnc-C18orf26-1 inhibited HSC proliferation and activation,downregulated TGF-β1-stimulatedα-SMA and COL1A2 expression,and inhibited the TGF-β1/Smad signaling pathway.HQD suppressed the proliferation and activation of HSCs.HQD increased miR-663a expression and decreased lnc-C18orf26-1 expression in HSCs.Further studies showed that HQD inhibited the expression of COL1A2,α-SMA,TGF-β1,TGF-βtype I receptor(TGF-βRI)and phosphorylated Smad2(p-Smad2)in HSCs,and these effects were reversed by miR-663a inhibitor treatment.Conclusion Our study identified lnc-C18orf26-1 and miR-663a as promising therapeutic targets for hepatic fibrosis.HQD inhibits HSC proliferation and activation at least partially by regulating the lnc-C18orf26-1/miR-663a/TGF-β1/TGF-βRI/p-Smad2 axis.
基金Project supported by the University Grants Commission,New Delhi
文摘The sorption study of La(Ⅲ) was carried out on poly[dibenzo-18-crown-6] and L-valine medium. The quantitative adsorption of La(Ⅲ) was found at 1×10-2 to 1×10-5 mol/L L-valine. The various eluting agents were found efficient eluents for La(Ⅲ). The capacity of crown polymer for La(Ⅲ) was found to be 0.43 ±0.01 mmol/g. The tolerance limit of various cations and anions for La(Ⅲ) was determined. La(Ⅲ) was quantitatively separated from other metal ions in binary as well as multicomponent mixtures. The study was extended to sequential separation of La(Ⅲ), U(VI) and Th(IV). The good separation yields were obtained and had good reproducibility (±2%). The method incorporated the determination of La(Ⅲ) in real sample. The method was simple, rapid and selective.
基金jointly supported by grants from the National Natural Science Foundation of China[Grant Nos.41576029 and 41690122(41690120)]the National Program on Global Change and Air–Sea Interaction(Grant No.GASIIPOVAI-03)+1 种基金the National Key Research and Development Program(Grant No.2018YFC1505802)the Strategic Priority Research Program of the Chinese Academy of Sciences(Grant Nos.XDA19060102 and XDB 40000000)。
文摘After the strong 2015/16 El Nino event,cold conditions prevailed in the tropical Pacific with the second-year cooling of the 2017/18 La Ni?a event.Many coupled models failed to predict the cold SST anomalies(SSTAs)in 2017.By using the ERA5 and GODAS(Global Ocean Data Assimilation System)products,atmospheric and oceanic factors were examined that could have been responsible for the second-year cooling,including surface wind and the subsurface thermal state.A time sequence is described to demonstrate how the cold SSTAs were produced in the central-eastern equatorial Pacific in late 2017.Since July 2017,easterly anomalies strengthened in the central Pacific;in the meantime,wind stress divergence anomalies emerged in the far eastern region,which strengthened during the following months and propagated westward,contributing to the development of the second-year cooling in 2017.At the subsurface,weak negative temperature anomalies were accompanied by upwelling in the eastern equatorial Pacific,which provided the cold water source for the sea surface.Thereafter,both the cold anomalies and upwelling were enhanced and extended westward in the centraleastern equatorial Pacific.These changes were associated with the seasonally weakened EUC(the Equatorial Undercurrent)and strengthened SEC(the South Equatorial Current),which favored more cold waters being accumulated in the central-equatorial Pacific.Then,the subsurface cold waters stretched upward with the convergence of the horizontal currents and eventually outcropped to the surface.The subsurface-induced SSTAs acted to induce local coupled air–sea interactions,which generated atmospheric–oceanic anomalies developing and evolving into the second-year cooling in the fall of 2017.
基金funded by the Major Research Plan of the National Natural Science Foundation of China(No.92159202)the National Key Research and Development Program of China(No.2021YFA1100500)+1 种基金the Leading Innovation Team Project of Hangzhou Medical College(No.CXLJ202401)the Key Research and Development Plan of Zhejiang Provincial Department of Science and Technology(No.2024C03051)。
文摘Objective:Cytotoxic T lymphocytes(CTLs)play a crucial role in the therapeutic approach to hepatocellular carcinoma(HCC).Recent research has indicated that junctional adhesion molecule-like protein(JAML)enhances the antitumor activity of CD8+T cells.Our study investigates the role of JAML+CD8+T cells in HCC.Methods:We utilized time-of-flight mass cytometry and an orthotopic mouse model of HCC to examine histone modifications in tumor-infiltrating immune cells undergoing immunotherapy.Flow cytometry was used to assess CD4+T cells differentiation and JAML expression in CD8+T cells infiltrating HCC.Correlation analysis revealed a strong positive correlation between lactate dehydrogenase A+(LDHA+)CD4+T cells and JAML+CD8+T cells.Subsequently,we evaluated the therapeutic effects of an agonistic anti-JAML antibody,both alone and combined with immunotherapy.Finally,RNA sequencing was conducted to identify potential regulatory mechanisms.Results:Immunotherapy significantly increased the percentage of CD8+T cells infiltrating HCC and induced histone modifications,such as H3K18 lactylation(H3K18la)in CD4+T cells.Flow cytometry analysis revealed that lactate promotes the differentiation of CD4+T cells into Th1 cells.LDHA,an enzyme that converts pyruvate to lactate,plays a key role in this process.Correlation analysis revealed a strong positive relationship between LDHA+CD4+T cells and JAML+CD8+T cells in patients who responded to immunotherapy.Moreover,high JAML expression in CD8+T cells was associated with a more favorable prognosis.In vivo experiments demonstrated that agonistic anti-JAML antibody therapy reduced tumor volume and significantly prolonged the survival of tumor-bearing mice,independent of the effects of anti-programmed cell death protein ligand-1 antibody(αPD-L1)-mediated immunotherapy.Pathway enrichment analysis further revealed that JAML enhances CTL responses through the oxidative phosphorylation pathway.Conclusions:Activation of JAML enhances CTL responses in HCC treatment,independent ofαPD-L1-mediated immunotherapy,providing a promising strategy for advanced HCC.
