AIM: To find the point mutations meaningful for inactivationof liver-related putative tumor suppressor gene (LPTS) gene,a human novel liver-related putative tumor suppressor geneand telomerase inhibitor in hepatocellu...AIM: To find the point mutations meaningful for inactivationof liver-related putative tumor suppressor gene (LPTS) gene,a human novel liver-related putative tumor suppressor geneand telomerase inhibitor in hepatocellular carcinoma.METHODS: The entire coding sequence of LPTS genewas examined for mutations by single strand conformationpolymorphism (SSCP) assay and PCR products directsequencing in 56 liver cancer cell lines, 7 ovarian cancerand 7 head & neck tumor cell lines and 70 pairs of HCCtissues samples. The cDNA fragment coding for the mostfrequent mutant protein was subcloned into GST fusionexpression vector. The product was expressed in E. coliand purified by glutathione-agarose column. Telomericrepeat amplification protocol (TRAP) assays wereperformed to study the effect of point mutation totelomerase inhibitory activity.RESULTS: SSCP gels showed the abnormal shifting bandsand DNA sequencing found that there were 5 differentmutations and/or polymorphisms in 12 tumor cell lineslocated at exon2, exon5 and exon7. The main alterationswere A(778)A/G and A(880)T in exon7. The change in siteof 778 could not be found in HCC tissue samples, while themutation in position 880 was seen in 7 (10 %) cases. Themutation in the site of 880 had no effect on telomeraseinhibitory activity.CONCLUSION: Alterations identified in this study arepolymorphisms of LPTS gene. LPTS mutations occur in HCCbut are infrequent and of little effect on the telomeraseinhibitory function of the protein. Epigenetics, such asmethylation, acetylation, may play the key role in inactivationof LPTS.展开更多
AIM: To analyze the expression levels of soluble form of CD95, CD95 ligand (sCD95 and sCD95L, respectively) in plasma and CD95 expression on CD3+ cells in livertransplanted recipients with acute rejection (AR).METHODS...AIM: To analyze the expression levels of soluble form of CD95, CD95 ligand (sCD95 and sCD95L, respectively) in plasma and CD95 expression on CD3+ cells in livertransplanted recipients with acute rejection (AR).METHODS: Peripheral blood mononuclear cells (PBMCs)were isolated from 30 clinically liver transplanted recipients.CD95 expression on CD3+ cells was quantitatively measured by two-color fluorescence activated cell sorter (FACS)analysis. Lymphocyte surface phenotypes of CD4, CD8,CD16 and CD55 were determined by flow cytometry.Plasma levels of sCD95 and sCD95L were detected byEnzyme Linked-Immuno-Sorbent Assay (ELISA). The results were compared with that from normal healthy volunteers (n = 15 individuals).RESULTS: FACS analysis showed that CD95 expression on CD3+ T cells was significantly increased in liver transplanted recipients with AR compared to that in stable recipients without rejection and infection or healthy individuals who did not undergo transplantation (18 676.93±11 588.34/molecule,6 848.20±1712.96/molecule, 6 418.01±2 001.95/molecule,respectively, P＜0.01). Whereas no significant difference was seen between liver-transplanted stable recipients and healthy individuals. Furthermore, no significant differences were detected between each group with CD4/CD8 ratio or the percentage of CD16+56+ cells. Plasma levels of sCD95were significantly higher in transplanted recipients with AR compered to that in stable recipients or healthy individuals (391.88±196.00, 201.37±30.30, 148.83±58.25 pg/mL,respectively, P＜0.01). In contrast, the plasma levels ofsCD95L in liver- transplanted recipients were not significantlydifferent from that in healthy individuals.CONCLUSION: The present results indicate that the increased CD95 expression on CD3+ cells and the increased levels of sCD95 in plasma may modify the immunological situation of the recipients after transplantation or represent the ongoing graft rejection.展开更多
Objective:Cancer stem cells(CSCs)have been the focus of several studies because oftheir involvement in cancer initiation and progression.CSCs were identified in 28%to 50%of hepatocellular carcinomas(HCCs).The origin o...Objective:Cancer stem cells(CSCs)have been the focus of several studies because oftheir involvement in cancer initiation and progression.CSCs were identified in 28%to 50%of hepatocellular carcinomas(HCCs).The origin of CSCs is still unclear,but it has been recently suggested that CSCs could originate from the transformation of liver progenitor cells(LPCs)during chronic liver inflammation.展开更多
AIM:To investigate the polymorphic simple sequencerepeat in intron 1 of the epidermal growth factor receptorgene(EGFR)(CA-SSRⅠ),which is known to affect theefficiency of gene transcription as a putative target of the...AIM:To investigate the polymorphic simple sequencerepeat in intron 1 of the epidermal growth factor receptorgene(EGFR)(CA-SSRⅠ),which is known to affect theefficiency of gene transcription as a putative target of themismatch repair(MMR)machinery in colorectal tumors.METHODS:The CA-SSRⅠgenotype was analyzed ina total of 86 primary colorectal tumors,selected upontheir microsatellite instability(MSI)status[42 with highfrequency MSI(MSI-H)and 44 microsatellite stable(MSS)]and their respective normal tissue.The effect of the CA-SSRⅠgenotype on the expression of the EGFR gene wasevaluated in 18 specimens using quantitative real-timereverse transcription PCR and immunohistochemistry.RESULTS:Mutations in CA-SSRⅠwere detected in 86%(36 of 42)of MSI-H colorectal tumors and 0%(0 of 44)ofMSS tumors,indicating the EGFR gene as a novel putativespecific target of the defective MMR system(P<0.001).Impaired expression of EGFR was detected in most ofthe colorectal tumors analyzed[6/12(50%)at the mRNAlevel and 15/18(83%)at the peptide level].However,noassociation was apparent between EGFR expression andCA-SSRⅠstatus in tumors or normal tissues.CONCLUSION:Our results suggest that CA-SSRⅠsequence does not contribute to the regulation of EGFRtranscription in colon,and should thus not be consideredas a promising predictive marker for response to EGFRinhibitors in patients with colorectal cancer.展开更多
Objective Fibrin deposition combined with glomerular epithelial cell proliferation and monocyte/macrophage infiltration is involved in the pathogenesis of crescentic glomerulonephritis and results from local thrombin ...Objective Fibrin deposition combined with glomerular epithelial cell proliferation and monocyte/macrophage infiltration is involved in the pathogenesis of crescentic glomerulonephritis and results from local thrombin generation.In addition to its effect on procoagulation and fibrinolysis,thrombin exerts many cellular effects through a functional thrombin receptor,protease-activated receptor-1(PAR-1).The paper is To determine the role of PAR-1 in the pathogenesis of crescentic glomerulonephritis.Methods wild type and PAR-1 knockout mice in an accelerated model of anti-glomerular basement menbrane antibody nephritis were studied.Results The wild type mice exhibited severe glomerular injury 10 days after injection of sheep anti-mouse glomerular besement membrane antibody,such as thrombus formation,extracapillary proiferation as well as renal failure.APR-1 knockout mice showed significant protection from crescentic glomerulonephritis.Conclusion PAR-1 plays a critical role in the pathogenesis of crescentic glomerulonephritis and may be a new target for therapeutic intervention.展开更多
Background For patients with stroke with large-vessel occlusion(LVO),study of factors predicting response to intravenous thrombolysis(IVT)would allow identifying subgroups with high expected gain,and those for whom it...Background For patients with stroke with large-vessel occlusion(LVO),study of factors predicting response to intravenous thrombolysis(IVT)would allow identifying subgroups with high expected gain,and those for whom it could be considered as futile,and even detrimental.From patients included in the Mechanical Thrombectomy After Intravenous Alteplase vs Alteplase Alone After Stroke trial,we investigated clinical-imaging factors associated with optimal response to IVT.Methods We included patients receiving IVT alone.Excellent outcome was defined by a 3-month modified Rankin Scale(mRS)score≤1.Clinical-imaging predictors were assessed on multivariate analysis after multiple imputations.The predictive performance of the model was assessed with the C-statistic.Results Among 247 patients with LVO treated with IVT alone,77(31%)showed 3-month mRS≤1.Predictors of 3-month mRS≤1 were no medical history of hypertension(OR 2.43;95%CI 1.74 to 3.38;p=0.007);no current smoking(OR 2.76;95%CI 1.79 to 4.26;p=0.02);onset-to IVT time(OR 0.47 per hour increase;95%CI 0.23 to 0.78;p=0.003);diffusion-weighted imaging(DWI)volume(OR 0.78 per 10 mL increase;95%CI 0.68 to 0.89;p=0.0004);presence of susceptibility vessel sign(SVS)(OR 7.89;95%CI 1.65 to 37.78;p=0.01)and SVS length(OR 0.87 per mm increase;95%CI 0.80 to 0.94;p=0.001).The prediction models showed a C-statistic=0.79(95%CI 0.79 to 0.80).Conclusions In patients with stroke with anterior-circulation LVO treated with IVT alone,predictors of excellent outcome at 3 months were no medical history of hypertension or current smoking,reduced onset-to IVT time,small DWI volume,presence of SVS and short SVS length.These predictive factors could help practitioners in decision-making for IVT implementation in reperfusion strategies,all the more for the drip and ship paradigm.Trial registration number NCT01062698.展开更多
基金a grant from National High Technology"863"Program of China,No.2001AA221021a grant from Special Funds for Major State Basic Research"973"of China,No.001CB510205+1 种基金a grant from the National Natural Sciences Foundation of China,No.30170524Chine-France PRA dans le domaine de la Biologie 2001.No.PRA B 01-05
文摘AIM: To find the point mutations meaningful for inactivationof liver-related putative tumor suppressor gene (LPTS) gene,a human novel liver-related putative tumor suppressor geneand telomerase inhibitor in hepatocellular carcinoma.METHODS: The entire coding sequence of LPTS genewas examined for mutations by single strand conformationpolymorphism (SSCP) assay and PCR products directsequencing in 56 liver cancer cell lines, 7 ovarian cancerand 7 head & neck tumor cell lines and 70 pairs of HCCtissues samples. The cDNA fragment coding for the mostfrequent mutant protein was subcloned into GST fusionexpression vector. The product was expressed in E. coliand purified by glutathione-agarose column. Telomericrepeat amplification protocol (TRAP) assays wereperformed to study the effect of point mutation totelomerase inhibitory activity.RESULTS: SSCP gels showed the abnormal shifting bandsand DNA sequencing found that there were 5 differentmutations and/or polymorphisms in 12 tumor cell lineslocated at exon2, exon5 and exon7. The main alterationswere A(778)A/G and A(880)T in exon7. The change in siteof 778 could not be found in HCC tissue samples, while themutation in position 880 was seen in 7 (10 %) cases. Themutation in the site of 880 had no effect on telomeraseinhibitory activity.CONCLUSION: Alterations identified in this study arepolymorphisms of LPTS gene. LPTS mutations occur in HCCbut are infrequent and of little effect on the telomeraseinhibitory function of the protein. Epigenetics, such asmethylation, acetylation, may play the key role in inactivationof LPTS.
文摘AIM: To analyze the expression levels of soluble form of CD95, CD95 ligand (sCD95 and sCD95L, respectively) in plasma and CD95 expression on CD3+ cells in livertransplanted recipients with acute rejection (AR).METHODS: Peripheral blood mononuclear cells (PBMCs)were isolated from 30 clinically liver transplanted recipients.CD95 expression on CD3+ cells was quantitatively measured by two-color fluorescence activated cell sorter (FACS)analysis. Lymphocyte surface phenotypes of CD4, CD8,CD16 and CD55 were determined by flow cytometry.Plasma levels of sCD95 and sCD95L were detected byEnzyme Linked-Immuno-Sorbent Assay (ELISA). The results were compared with that from normal healthy volunteers (n = 15 individuals).RESULTS: FACS analysis showed that CD95 expression on CD3+ T cells was significantly increased in liver transplanted recipients with AR compared to that in stable recipients without rejection and infection or healthy individuals who did not undergo transplantation (18 676.93±11 588.34/molecule,6 848.20±1712.96/molecule, 6 418.01±2 001.95/molecule,respectively, P＜0.01). Whereas no significant difference was seen between liver-transplanted stable recipients and healthy individuals. Furthermore, no significant differences were detected between each group with CD4/CD8 ratio or the percentage of CD16+56+ cells. Plasma levels of sCD95were significantly higher in transplanted recipients with AR compered to that in stable recipients or healthy individuals (391.88±196.00, 201.37±30.30, 148.83±58.25 pg/mL,respectively, P＜0.01). In contrast, the plasma levels ofsCD95L in liver- transplanted recipients were not significantlydifferent from that in healthy individuals.CONCLUSION: The present results indicate that the increased CD95 expression on CD3+ cells and the increased levels of sCD95 in plasma may modify the immunological situation of the recipients after transplantation or represent the ongoing graft rejection.
文摘Objective:Cancer stem cells(CSCs)have been the focus of several studies because oftheir involvement in cancer initiation and progression.CSCs were identified in 28%to 50%of hepatocellular carcinomas(HCCs).The origin of CSCs is still unclear,but it has been recently suggested that CSCs could originate from the transformation of liver progenitor cells(LPCs)during chronic liver inflammation.
文摘AIM:To investigate the polymorphic simple sequencerepeat in intron 1 of the epidermal growth factor receptorgene(EGFR)(CA-SSRⅠ),which is known to affect theefficiency of gene transcription as a putative target of themismatch repair(MMR)machinery in colorectal tumors.METHODS:The CA-SSRⅠgenotype was analyzed ina total of 86 primary colorectal tumors,selected upontheir microsatellite instability(MSI)status[42 with highfrequency MSI(MSI-H)and 44 microsatellite stable(MSS)]and their respective normal tissue.The effect of the CA-SSRⅠgenotype on the expression of the EGFR gene wasevaluated in 18 specimens using quantitative real-timereverse transcription PCR and immunohistochemistry.RESULTS:Mutations in CA-SSRⅠwere detected in 86%(36 of 42)of MSI-H colorectal tumors and 0%(0 of 44)ofMSS tumors,indicating the EGFR gene as a novel putativespecific target of the defective MMR system(P<0.001).Impaired expression of EGFR was detected in most ofthe colorectal tumors analyzed[6/12(50%)at the mRNAlevel and 15/18(83%)at the peptide level].However,noassociation was apparent between EGFR expression andCA-SSRⅠstatus in tumors or normal tissues.CONCLUSION:Our results suggest that CA-SSRⅠsequence does not contribute to the regulation of EGFRtranscription in colon,and should thus not be consideredas a promising predictive marker for response to EGFRinhibitors in patients with colorectal cancer.
文摘Objective Fibrin deposition combined with glomerular epithelial cell proliferation and monocyte/macrophage infiltration is involved in the pathogenesis of crescentic glomerulonephritis and results from local thrombin generation.In addition to its effect on procoagulation and fibrinolysis,thrombin exerts many cellular effects through a functional thrombin receptor,protease-activated receptor-1(PAR-1).The paper is To determine the role of PAR-1 in the pathogenesis of crescentic glomerulonephritis.Methods wild type and PAR-1 knockout mice in an accelerated model of anti-glomerular basement menbrane antibody nephritis were studied.Results The wild type mice exhibited severe glomerular injury 10 days after injection of sheep anti-mouse glomerular besement membrane antibody,such as thrombus formation,extracapillary proiferation as well as renal failure.APR-1 knockout mice showed significant protection from crescentic glomerulonephritis.Conclusion PAR-1 plays a critical role in the pathogenesis of crescentic glomerulonephritis and may be a new target for therapeutic intervention.
文摘Background For patients with stroke with large-vessel occlusion(LVO),study of factors predicting response to intravenous thrombolysis(IVT)would allow identifying subgroups with high expected gain,and those for whom it could be considered as futile,and even detrimental.From patients included in the Mechanical Thrombectomy After Intravenous Alteplase vs Alteplase Alone After Stroke trial,we investigated clinical-imaging factors associated with optimal response to IVT.Methods We included patients receiving IVT alone.Excellent outcome was defined by a 3-month modified Rankin Scale(mRS)score≤1.Clinical-imaging predictors were assessed on multivariate analysis after multiple imputations.The predictive performance of the model was assessed with the C-statistic.Results Among 247 patients with LVO treated with IVT alone,77(31%)showed 3-month mRS≤1.Predictors of 3-month mRS≤1 were no medical history of hypertension(OR 2.43;95%CI 1.74 to 3.38;p=0.007);no current smoking(OR 2.76;95%CI 1.79 to 4.26;p=0.02);onset-to IVT time(OR 0.47 per hour increase;95%CI 0.23 to 0.78;p=0.003);diffusion-weighted imaging(DWI)volume(OR 0.78 per 10 mL increase;95%CI 0.68 to 0.89;p=0.0004);presence of susceptibility vessel sign(SVS)(OR 7.89;95%CI 1.65 to 37.78;p=0.01)and SVS length(OR 0.87 per mm increase;95%CI 0.80 to 0.94;p=0.001).The prediction models showed a C-statistic=0.79(95%CI 0.79 to 0.80).Conclusions In patients with stroke with anterior-circulation LVO treated with IVT alone,predictors of excellent outcome at 3 months were no medical history of hypertension or current smoking,reduced onset-to IVT time,small DWI volume,presence of SVS and short SVS length.These predictive factors could help practitioners in decision-making for IVT implementation in reperfusion strategies,all the more for the drip and ship paradigm.Trial registration number NCT01062698.
