AIM:To explore the role of high-mobility group box 1 (HMGB1) protein during liver fibrogenesis and investigate the functional effects of HMGB1 gene silencing in hepatic stellate cells (HSCs) using siRNA.METHODS:Hepati...AIM:To explore the role of high-mobility group box 1 (HMGB1) protein during liver fibrogenesis and investigate the functional effects of HMGB1 gene silencing in hepatic stellate cells (HSCs) using siRNA.METHODS:Hepatic fibrosis in rats was induced through serial subcutaneous injections of dimethylnitrosamine,and expression of HMGB1 was detected by immunohistochemistry.HMGB1 siRNAs were developed and transiently transfected into HSC-T6 cells using Lipofectamine 2000.HMGB1 expression was evaluated by real-time polymerase chain reaction (PCR) and Western blotting analysis.Expression of α-smooth muscle actin (α-SMA) and collagen typesⅠand Ⅲ was evaluated by real-time PCR.Cell proliferation and the cell cycle were determined using the methyl thiazolyl tetrazolium method.Finally,collagen content in HSC supernatant was evaluated by an enzyme-linked immunosorbent assay.RESULTS:The results showed that HMGB1 was upregulated during liver fibrosis and that its expression was closely correlated with the deposition of collagen.siRNA molecules were successfully transfected into HSCs and induced inhibition of HMGB1 expression in a time-dependent manner.Moreover,HMGB1 siRNA treatment inhibited synthesis of α-SMA and collagen types Ⅰ and Ⅲ in transfected HSCs.CONCLUSION:This study suggests a significant functional role for HMGB1 in the development of liver fibrosis.It also demonstrates that downregulation of HMGB1 expression might be a potential strategy to treat liver fibrosis.展开更多
The influence of cells groupings factor to the performance of the cells groupings time-shift pilot scheme is researched for the multiple cells large scale antennas systems（LSAS）. The former researches have confirmed...The influence of cells groupings factor to the performance of the cells groupings time-shift pilot scheme is researched for the multiple cells large scale antennas systems（LSAS）. The former researches have confirmed that the cells groupings time-shift pilots scheme is effective to reduce inter-cell interference, especially pilot contamination, which results from the pilot reuse in adjacent cells. However, they have not specified reasonable cells groupings factor, which plays a critical role in the general performance of the LSAS. Therefore, this problem is researched in details. The time for reverse-link data transmission will be compressed, when the groupings factor surpasses a certain range. Thus it is not always beneficial to increase the cells groupings factor without limitation. Furthermore,a reasonable cells groupings factor is deduced from the perspective of optimization to enhance the system performance. Simulations verify the proposed cell grouping factor.展开更多
BACKGROUND: Liver failure in chronic hepatitis B (CHB) patients is a severe, life-threatening condition. Intestinal endotoxemia plays a significant role in the progress to liver failure. High mobility group box-1 (HMG...BACKGROUND: Liver failure in chronic hepatitis B (CHB) patients is a severe, life-threatening condition. Intestinal endotoxemia plays a significant role in the progress to liver failure. High mobility group box-1 (HMGB1) protein is involved in the process of endotoxemia. Regulatory T (Treg) cells maintain immune tolerance and contribute to the immunological hyporesponsiveness against HBV infection. However, the roles of HMGB1 and Treg cells in the pathogenesis of liver failure in CHB patients, and whether HMGB1 affects the immune activity of Treg cells are poorly known at present, and so were explored in this study. METHODS: The levels of HMGB1 expression were detected by ELISA, real-time RT-PCR, and Western blotting, and the percentage of CD4(+)CD25(+)CD127(low) Treg cells among CD4(+) cells was detected by flow cytometry in liver failure patients with chronic HBV infection, CHB patients, and healthy controls. Then, CD4(+)CD25(+)CD127(low) Treg cells isolated from the peripheral blood mononuclear cells from CHB patients were stimulated with HMGB1 at different concentrations or at various intervals. The effect of HMGB1 on the immune activity of Treg cells was assessed by a suppression assay of the allogeneic mixed lymphocyte response. The levels of forkhead box P3 (Foxp3) expression in Treg cells treated with HMGB1 were detected by RT-PCR and Western blotting. RESULTS: A higher level of HMGB1 expression and a lower percentage of Treg cells within the population of CIA(+) cells were found in liver failure patients than in CHB patients (82.6+/-20.1 mu g/L vs. 34.2+/-13.7 mu g/L; 4.55+/-1.34% vs. 9.52+/-3.89%, respectively). The immune activity of Treg cells was significantly weakened and the levels of Foxp3 expression were reduced in a dose- or time-dependent manner when Treg cells were stimulated with HMGB1 in vitro. CONCLUSIONS: The high level of HMGB1 and the low percentage of Treg cells play an important role in the pathogenesis of liver failure in patients with chronic HBV infection. Moreover, HMGB1 can weaken the immune activity of Treg cells. It is suggested that effectively inhibiting HMGB1 expression could be a feasible way to treat liver failure by suppressing endotoxemia and enhancing Treg cell activity.展开更多
AIM:To investigate the effect of high mobility group A2(HMGA2) gene silencing on gastric cancer MKN-45 cells in vitro.METHODS:HMGA2 short hairpin RNA(shRNA) expression plasmids were constructed,including a pair of ran...AIM:To investigate the effect of high mobility group A2(HMGA2) gene silencing on gastric cancer MKN-45 cells in vitro.METHODS:HMGA2 short hairpin RNA(shRNA) expression plasmids were constructed,including a pair of random scrambled sequences.Human gastric cancer cell line MKN-45 cells were divided into three groups:blank control group(non-transfected cells),transfected group(cells transfected with HMGA2 shRNA recombinant plasmid) and scrambled sequence group(transfected with random scrambled plasmid).Cells were transfected with HMGA2 shRNA recombinant plasmids and scrambled plasmid in vitro,and the cells transfection efficiency was assayed by fluorescence microscopy.The HMGA2 messenger RNA(mRNA) expression was detected by reverse transcription polymerase chain reaction,gastric cancer cells apoptosis was detected by flow cytometry,cell proliferation was detected by methyl thiazol tetrazolium,and the protein expression of phosphatidylinositol 3-kinase(PI3K),protein kinase B(Akt),P27,caspase-9 and B-cell leukemia/lymphoma-2(Bcl-2) were analyzed by Western blotting.RESULTS:Compared with the blank control group and the scrambled sequence group,the levels of HMGA2 mRNA and protein expression in the transfected group were significantly reduced(P < 0.05).The relative HMGA2 mRNA expression levels of the blank control group,transfected group and scrambled sequence group were 0.674 ± 0.129,0.374 ± 0.048 and 0.689 ± 0.124,respectively.The relative HMGA2 protein expression levels of the blank control group,transfected group and scrambled sequence group were 0.554 ± 0.082,0.113 ± 0.032 and 0.484 ± 0.123,respectively.Moreover,transfection with the scrambled sequence had no effect on the expression of HMGA2.After being transfected with shRNA for 24,48 and 72 h,the cell apoptotic rates of the transfected group were 21.65% ± 0.28%,39.98% ± 1.82% and 24.51% ± 0.93%,respectively,which significantly higher than those of blank control group(4.72% ± 1.34%,5.83% ± 0.13% and 5.22% ± 1.07%) and scrambled sequence group(4.28% ± 1.33%,7.87% ± 1.43% and 6.71% ± 0.92%).After 24,48 and 72 h,the cell proliferation inhibition rates in the transfected group were 31.57% ± 1.17%,39.45% ± 2.07% and 37.56% ± 2.32%,respectively;the most obvious cell proliferation inhibition appeared at 48 h after transfection.Compared with the blank control group and scrambled sequence group,after transfection of shRNA for 72 h,the protein expression levels of PI3K(0.042 ± 0.005 vs 0.069 ± 0.003,0.067 ± 0.05),Akt(0.248 ± 0.004 vs 0.489 ± 0.006,0.496 ± 0.104) and Bcl-2(0.295 ± 0.084 vs 0.592 ± 0.072,0.594 ± 0.109) were significantly reduced.The protein expression levels of P27(0.151 ± 0.010 vs 0.068± 0.014,0.060 ± 0.013) and caspase-9(0.136 ± 0.042 vs 0.075 ± 0.010,0.073 ± 0.072) were significantly upregulated.CONCLUSION:HMGA2 shRNA gene silencing induces apoptosis and suppresses proliferation of MKN-45 cells.展开更多
Objective To investigate the expression of Snail in bladder urothelial carcinoma and evaluate its relationship with E-cadherin and a subset of T cell groups. Methods Immunohistochemical method was used to detect the e...Objective To investigate the expression of Snail in bladder urothelial carcinoma and evaluate its relationship with E-cadherin and a subset of T cell groups. Methods Immunohistochemical method was used to detect the expression of Snail and E-cadherin proteins in tissue展开更多
High mobility group box-1 protein(HMGB1),which is a nuclear protein,participates in chromatin architecture and transcriptional regulation.When released from cells,HMGB1 also plays a well-established role as a pro-infl...High mobility group box-1 protein(HMGB1),which is a nuclear protein,participates in chromatin architecture and transcriptional regulation.When released from cells,HMGB1 also plays a well-established role as a pro-inflammatory mediator during innate immune responses to injury.In the initial stage of injury,there is a release of large quantities of early pro-inflammatory mediators to initiate or perpetuate immune responses against pathogens,but this pro-inflammatory period is transient,and it is followed by a prolonged period of immune suppression.At present,several lines of evidences have suggested that HMGB1 is a late cytokine provoking delayed endotoxin morbidity,which may enhance the production of early proinflammatory mediators,and it can contribute potently to the activation of different immune cells and play a role in the development of host cell-mediated immunity.The biology of HMGB1 has been extensively studied as a pro-inflammatory cytokine of systemic inflammation,however,this review will attempt to provide a summary of the effects of HMGB1 on different immune cells and its regulatory mechanism in acute insults.展开更多
The purpose of this experiment is to prepare a kind of large colored microspheres which can be observed under optical microscope and used for cell labelling. In the presence of organic solvent, electrolyte and dye, mi...The purpose of this experiment is to prepare a kind of large colored microspheres which can be observed under optical microscope and used for cell labelling. In the presence of organic solvent, electrolyte and dye, microspheres [styrene/acrylamide/dye (St AAm-Dye ) copolymerJ were synthesized from styrene and acrylamide by means of emulsifier-free emulsion polymerization. The carboxyl groups on the surface were derived from acrylarnide by hydrazinolysis and carboxylate reaction. The diameters of microspheres and the content of carboxyl groups on the surface were measured. The effect of the polymerization condition on microsphere size was discussed. The diam6ter of the colored microsphere was 3±0. 05μm; the content of carboxyl groups on the surface was 190. 5μmol/g (dry solids). The results indicate that it would be possible to obtain monodisperse and colored large polymer microspheres by this reported method.展开更多
基金Supported by The Select and Train Outstanding Young Teach-ers Foundation of Shanghai,No.jdy08086WUJieping Experimental Diagnosis of Liver Disease Medical Foundation,No.LDWMF-SY-2011B009
文摘AIM:To explore the role of high-mobility group box 1 (HMGB1) protein during liver fibrogenesis and investigate the functional effects of HMGB1 gene silencing in hepatic stellate cells (HSCs) using siRNA.METHODS:Hepatic fibrosis in rats was induced through serial subcutaneous injections of dimethylnitrosamine,and expression of HMGB1 was detected by immunohistochemistry.HMGB1 siRNAs were developed and transiently transfected into HSC-T6 cells using Lipofectamine 2000.HMGB1 expression was evaluated by real-time polymerase chain reaction (PCR) and Western blotting analysis.Expression of α-smooth muscle actin (α-SMA) and collagen typesⅠand Ⅲ was evaluated by real-time PCR.Cell proliferation and the cell cycle were determined using the methyl thiazolyl tetrazolium method.Finally,collagen content in HSC supernatant was evaluated by an enzyme-linked immunosorbent assay.RESULTS:The results showed that HMGB1 was upregulated during liver fibrosis and that its expression was closely correlated with the deposition of collagen.siRNA molecules were successfully transfected into HSCs and induced inhibition of HMGB1 expression in a time-dependent manner.Moreover,HMGB1 siRNA treatment inhibited synthesis of α-SMA and collagen types Ⅰ and Ⅲ in transfected HSCs.CONCLUSION:This study suggests a significant functional role for HMGB1 in the development of liver fibrosis.It also demonstrates that downregulation of HMGB1 expression might be a potential strategy to treat liver fibrosis.
基金supported by the National Natural Science Foundation of China(6110602261574013)
文摘The influence of cells groupings factor to the performance of the cells groupings time-shift pilot scheme is researched for the multiple cells large scale antennas systems（LSAS）. The former researches have confirmed that the cells groupings time-shift pilots scheme is effective to reduce inter-cell interference, especially pilot contamination, which results from the pilot reuse in adjacent cells. However, they have not specified reasonable cells groupings factor, which plays a critical role in the general performance of the LSAS. Therefore, this problem is researched in details. The time for reverse-link data transmission will be compressed, when the groupings factor surpasses a certain range. Thus it is not always beneficial to increase the cells groupings factor without limitation. Furthermore,a reasonable cells groupings factor is deduced from the perspective of optimization to enhance the system performance. Simulations verify the proposed cell grouping factor.
基金supported by a grant from the National Natural Science Foundation of China (No. 81071342)
文摘BACKGROUND: Liver failure in chronic hepatitis B (CHB) patients is a severe, life-threatening condition. Intestinal endotoxemia plays a significant role in the progress to liver failure. High mobility group box-1 (HMGB1) protein is involved in the process of endotoxemia. Regulatory T (Treg) cells maintain immune tolerance and contribute to the immunological hyporesponsiveness against HBV infection. However, the roles of HMGB1 and Treg cells in the pathogenesis of liver failure in CHB patients, and whether HMGB1 affects the immune activity of Treg cells are poorly known at present, and so were explored in this study. METHODS: The levels of HMGB1 expression were detected by ELISA, real-time RT-PCR, and Western blotting, and the percentage of CD4(+)CD25(+)CD127(low) Treg cells among CD4(+) cells was detected by flow cytometry in liver failure patients with chronic HBV infection, CHB patients, and healthy controls. Then, CD4(+)CD25(+)CD127(low) Treg cells isolated from the peripheral blood mononuclear cells from CHB patients were stimulated with HMGB1 at different concentrations or at various intervals. The effect of HMGB1 on the immune activity of Treg cells was assessed by a suppression assay of the allogeneic mixed lymphocyte response. The levels of forkhead box P3 (Foxp3) expression in Treg cells treated with HMGB1 were detected by RT-PCR and Western blotting. RESULTS: A higher level of HMGB1 expression and a lower percentage of Treg cells within the population of CIA(+) cells were found in liver failure patients than in CHB patients (82.6+/-20.1 mu g/L vs. 34.2+/-13.7 mu g/L; 4.55+/-1.34% vs. 9.52+/-3.89%, respectively). The immune activity of Treg cells was significantly weakened and the levels of Foxp3 expression were reduced in a dose- or time-dependent manner when Treg cells were stimulated with HMGB1 in vitro. CONCLUSIONS: The high level of HMGB1 and the low percentage of Treg cells play an important role in the pathogenesis of liver failure in patients with chronic HBV infection. Moreover, HMGB1 can weaken the immune activity of Treg cells. It is suggested that effectively inhibiting HMGB1 expression could be a feasible way to treat liver failure by suppressing endotoxemia and enhancing Treg cell activity.
基金Supported by The Natural Science Foundation of Guangxi,No. 2010GXNSFA013166the Key Project of Health Department of Guangxi,No.Zhong 2010021
文摘AIM:To investigate the effect of high mobility group A2(HMGA2) gene silencing on gastric cancer MKN-45 cells in vitro.METHODS:HMGA2 short hairpin RNA(shRNA) expression plasmids were constructed,including a pair of random scrambled sequences.Human gastric cancer cell line MKN-45 cells were divided into three groups:blank control group(non-transfected cells),transfected group(cells transfected with HMGA2 shRNA recombinant plasmid) and scrambled sequence group(transfected with random scrambled plasmid).Cells were transfected with HMGA2 shRNA recombinant plasmids and scrambled plasmid in vitro,and the cells transfection efficiency was assayed by fluorescence microscopy.The HMGA2 messenger RNA(mRNA) expression was detected by reverse transcription polymerase chain reaction,gastric cancer cells apoptosis was detected by flow cytometry,cell proliferation was detected by methyl thiazol tetrazolium,and the protein expression of phosphatidylinositol 3-kinase(PI3K),protein kinase B(Akt),P27,caspase-9 and B-cell leukemia/lymphoma-2(Bcl-2) were analyzed by Western blotting.RESULTS:Compared with the blank control group and the scrambled sequence group,the levels of HMGA2 mRNA and protein expression in the transfected group were significantly reduced(P < 0.05).The relative HMGA2 mRNA expression levels of the blank control group,transfected group and scrambled sequence group were 0.674 ± 0.129,0.374 ± 0.048 and 0.689 ± 0.124,respectively.The relative HMGA2 protein expression levels of the blank control group,transfected group and scrambled sequence group were 0.554 ± 0.082,0.113 ± 0.032 and 0.484 ± 0.123,respectively.Moreover,transfection with the scrambled sequence had no effect on the expression of HMGA2.After being transfected with shRNA for 24,48 and 72 h,the cell apoptotic rates of the transfected group were 21.65% ± 0.28%,39.98% ± 1.82% and 24.51% ± 0.93%,respectively,which significantly higher than those of blank control group(4.72% ± 1.34%,5.83% ± 0.13% and 5.22% ± 1.07%) and scrambled sequence group(4.28% ± 1.33%,7.87% ± 1.43% and 6.71% ± 0.92%).After 24,48 and 72 h,the cell proliferation inhibition rates in the transfected group were 31.57% ± 1.17%,39.45% ± 2.07% and 37.56% ± 2.32%,respectively;the most obvious cell proliferation inhibition appeared at 48 h after transfection.Compared with the blank control group and scrambled sequence group,after transfection of shRNA for 72 h,the protein expression levels of PI3K(0.042 ± 0.005 vs 0.069 ± 0.003,0.067 ± 0.05),Akt(0.248 ± 0.004 vs 0.489 ± 0.006,0.496 ± 0.104) and Bcl-2(0.295 ± 0.084 vs 0.592 ± 0.072,0.594 ± 0.109) were significantly reduced.The protein expression levels of P27(0.151 ± 0.010 vs 0.068± 0.014,0.060 ± 0.013) and caspase-9(0.136 ± 0.042 vs 0.075 ± 0.010,0.073 ± 0.072) were significantly upregulated.CONCLUSION:HMGA2 shRNA gene silencing induces apoptosis and suppresses proliferation of MKN-45 cells.
文摘Objective To investigate the expression of Snail in bladder urothelial carcinoma and evaluate its relationship with E-cadherin and a subset of T cell groups. Methods Immunohistochemical method was used to detect the expression of Snail and E-cadherin proteins in tissue
基金supported,in part,by grants from the National Natural Science Foundation(81130035,30971192,81071545,81272090,81121004)the National Basic Research Program of China(2012CB518102)
文摘High mobility group box-1 protein(HMGB1),which is a nuclear protein,participates in chromatin architecture and transcriptional regulation.When released from cells,HMGB1 also plays a well-established role as a pro-inflammatory mediator during innate immune responses to injury.In the initial stage of injury,there is a release of large quantities of early pro-inflammatory mediators to initiate or perpetuate immune responses against pathogens,but this pro-inflammatory period is transient,and it is followed by a prolonged period of immune suppression.At present,several lines of evidences have suggested that HMGB1 is a late cytokine provoking delayed endotoxin morbidity,which may enhance the production of early proinflammatory mediators,and it can contribute potently to the activation of different immune cells and play a role in the development of host cell-mediated immunity.The biology of HMGB1 has been extensively studied as a pro-inflammatory cytokine of systemic inflammation,however,this review will attempt to provide a summary of the effects of HMGB1 on different immune cells and its regulatory mechanism in acute insults.
文摘The purpose of this experiment is to prepare a kind of large colored microspheres which can be observed under optical microscope and used for cell labelling. In the presence of organic solvent, electrolyte and dye, microspheres [styrene/acrylamide/dye (St AAm-Dye ) copolymerJ were synthesized from styrene and acrylamide by means of emulsifier-free emulsion polymerization. The carboxyl groups on the surface were derived from acrylarnide by hydrazinolysis and carboxylate reaction. The diameters of microspheres and the content of carboxyl groups on the surface were measured. The effect of the polymerization condition on microsphere size was discussed. The diam6ter of the colored microsphere was 3±0. 05μm; the content of carboxyl groups on the surface was 190. 5μmol/g (dry solids). The results indicate that it would be possible to obtain monodisperse and colored large polymer microspheres by this reported method.
