AIM： To study the relationship between interleukin-lbeta （IL-1β） up-regulating tissue inhibitor of matrix metalloproteinase-1 （TIMMP-1） mRNA expression and phosphorylation of both c-jun N-terminal kinase （INK）...AIM： To study the relationship between interleukin-lbeta （IL-1β） up-regulating tissue inhibitor of matrix metalloproteinase-1 （TIMMP-1） mRNA expression and phosphorylation of both c-jun N-terminal kinase （INK） and p38 in rat heffatic stellate cells （HSC）. METHODS： RT-PCR was performed to measure the expression of TIMMP-1 mRNA in rat HSC. Western blot was performed to measure IL-1β-induced JNK and p38 activities in rat HSC. RESULTS： TIMMP-1 mRNA expression （1.191± 0.079） was much higher after treatment with IL-1β （10 ng/mL） for 24 h than in control group （0.545±0.091） （P〈0.01）. IL-1β activated INK and p38 in a time-dependent manner. After stimulation with IL-1β for 0, 5, 15, 30, 60 and 120 min, the INK activity was 0.982±0.299, 1.501±0.720, 2.133±0.882, 3.360±0.452, 2.181±0.789, and 1.385 ± 0.368, respectively. There was a significant difference in JNK activity at 15 min （P〈 0.01）, 30 min （P〈 0.01） and 60 min （P〈0.01） in comparison to that at 0 min. The p38 activity was 1.061±0.310, 2.050±0.863, 2.380±0.573, 2.973±0.953, 2.421±0.793, and 1.755 ± 0.433 at the 6 time points （0, 5, 15, 30, 60 and 120 min） respectively. There was a significant difference in p38 activity at 5 min （P〈0.05）, 15 min （P〈0.01）, 30 min （P〈0.01） and 60 min （P〈0.01） compared to that at 0 min. TIMMP-1 mRNA expression trended to decrease in 3 groups pretreated with different concentrations of SP600125 （10 μmol/L, 1.022±0.113; 20 μmol/L, 0.869±0.070; 40 μmol/L, 0.666±0.123）. Their decreases were all significant （P〈0.05, P〈0.01, P〈0.01） in comparison to control group （without SP600125 treatment, 1.163±0.107）. In the other 3 groups pretreated with different concentrations of SB203580 （10 μmol/L, 1.507±0.099; 20 μmol/L, 1.698±0.107; 40 μmol/L, 1.857±0.054）, the expression of TIMMP-1 mRNA increased. Their levels were higher than those in the control group （without SB203580 treatment, 1.027 ± 0.061） with a si展开更多
Under normal conditions, the sympathetic neurotransmitter noradrenaline inhibits the production and release of pro-inflammatory cytokines. However, after peripheral nerve and tissue injury, pro-inflammatory cytokines ...Under normal conditions, the sympathetic neurotransmitter noradrenaline inhibits the production and release of pro-inflammatory cytokines. However, after peripheral nerve and tissue injury, pro-inflammatory cytokines appear to induce the expression of the alphalA-adreno- ceptor subtype on immune cells and perhaps also on other cells in the injured tissue. In turn, noradrenaline may act on up-regulated alphal-adrenoceptors to increase the production of the pro-inflammatory cytokine interleukin-6. In addition, the release of inflammatory mediators and nerve growth factor from keratinocytes and other cells may augment the expression of alphal-adrenoceptors on peripheral nerve fibers. Consequently, nociceptive afferents acquire an abnormal excitability to adrenergic agents, and inflammatory processes build. These mechanisms could contribute to the development of sympathetically maintained pain in conditions such as post-herpetic neuralgia, cutaneous neuromas, amputation stump pain and complex regional pain syndrome.展开更多
Although prohibitin 1(PHB) is known to be associated with tumors,there are few studies regarding the role of PHB in hepatocellular carcinoma.An earlier glycoproteomics study of ours showed that PHB is over-expressed i...Although prohibitin 1(PHB) is known to be associated with tumors,there are few studies regarding the role of PHB in hepatocellular carcinoma.An earlier glycoproteomics study of ours showed that PHB is over-expressed in MHCC97-H cells,a highly metastatic liver cancer cell line,and can be captured by wheat germ agglutinin.In the present study,western blotting and reverse transcription-PCR experiments showed an approximately 2-fold up-regulation in the expression of PHB in MHCC97-H cells.However,PHB was not significantly up-regulated in MHCC97-L cells,a low-metastatic liver cancer cell line.When PHB was over-expressed in MHCC97-L cells,cell proliferation was inhibited by 35% and migration increased about 2-fold.The results of this study show that PHB is up-regulated in MHCC97-H cells and is associated with both proliferation and migration of liver cancer cells.展开更多
AIM:To investigate cytokeratin 8(CK8)overexpression during hepatitis C virus(HCV)infection and its pathogenesis,and the effect of ectopic CK8 expression on hepatoma cell lines.METHODS:We successfully established an in...AIM:To investigate cytokeratin 8(CK8)overexpression during hepatitis C virus(HCV)infection and its pathogenesis,and the effect of ectopic CK8 expression on hepatoma cell lines.METHODS:We successfully established an in vitro HCV cell culture system(HCVcc)to investigate the different expression profiles of CK8 in Huh-7-HCV and Huh-7.5-HCV cells.The expression of CK8 at the mRNA level was determined by real-time polymerase chain reaction(RT-PCR).The expression of CK8 at the protein level was evaluated by Western blotting.We then constructed a eukaryotic expression combination vector containing the coding sequence of human full length CK8 gene.CK8cDNA was amplified by reverse transcription-PCR and inserted into pEGFP-C1 and the positive clone pEGFPCK8 was obtained.After confirming the sequence,the recombinant plasmid was transfected into SMMC7721cells with lipofectamine2000 and CK8 expression was detected using inverted fluorescence microscopy,RTPCR and Western blotting.Besides,we identified biological function of CK8 on SMMC7721 cells,including cell proliferation,cell cycle and apoptosis detection.RESULTS:RT-PCR showed that the expression level of CK8 in Huh-7-HCV and Huh-7.5-HCV cells was 2.88and 2.95 times higher than in control cells.Western blot showed that CK8 expression in Huh-7-HCV and Huh-7.5-HCV cells was 2.53 and 3.26 times higher than that in control cells,respectively.We found that CK8at mRNA and protein levels were both significantly increased in HCVcc.CK8 was up-regulated in SMMC7721cells.CK8 expression at the mRNA level was significantly upregulated in SMMC7721/pEGFP-CK8 cells.CK8expression in SMMC7721/pEGFP-CK8 cells was 2.69times higher than in SMMC7721 cells,and was 2.64times higher than in SMMC7721/pEGFP-C1 cells.CK8expression at the protein level in SMMC7721/pEGFPCK8 cells was 2.46 times higher than in SMMC7721cells,and was 2.29 times higher than in SMMC7721/pEGFP-C1 cells.Further analysis demonstrated that forced expression of CK8 slowed cell growth and induced apoptosis of SMMC7721 cells.CONCLU展开更多
OBJECTIVE: To investigate the expression of Toll-like receptor 2 (TLR2) and Toll-like receptor 4 (TLR4) and the effect of lipopolysaccharide (LPS) on their expression in cultured endothelial cells. METHODS: Total RNA ...OBJECTIVE: To investigate the expression of Toll-like receptor 2 (TLR2) and Toll-like receptor 4 (TLR4) and the effect of lipopolysaccharide (LPS) on their expression in cultured endothelial cells. METHODS: Total RNA was extracted from ECV304 cells and isolated human umbilical vein endothelial cells (HUVECs) exposed to LPS, respectively. The quantification of TLR2 and TLR4 mRNA in HUVECs and EVC304 cells was carried out by reverse transcriptase-polymerase chain reaction (RT-PCR). RESULTS: ECV304 cells and HUVECs were able to express TLR2 and TLR4 mRNA, but the expression levels of TLR4 appeared to be stronger than those of TLR2. LPS could upregulate the expression levels of TLR4 obviously, whereas it had no effect on the expression level of TLR2. CONCLUSIONS: Our data indicate that TLR4 may be the LPS signal transducer in endothelial cells and plays important roles in the cell activation of LPS. The ECV304 cell line is a better experimental model than isolated HUVECs in the research of endothelial cells.展开更多
Dear Editor,As the key contributor to the rising phase of action potentials in dorsal root ganglion（DRG）neurons,voltagegated sodium channels（VGSCs）are important in physiological and pathological pain conditions.Fo...Dear Editor,As the key contributor to the rising phase of action potentials in dorsal root ganglion（DRG）neurons,voltagegated sodium channels（VGSCs）are important in physiological and pathological pain conditions.For instance,abnormal expression of VGSCs in DRG neurons is the展开更多
The Asian swamp eel （Monopterus albus） is one of the most economically important freshwater fish in East Asia, but data on the immune genes of M. albus are scarce compared to other commercially important fish. A bet...The Asian swamp eel （Monopterus albus） is one of the most economically important freshwater fish in East Asia, but data on the immune genes of M. albus are scarce compared to other commercially important fish. A better understanding of the eel＇s immune responses may help in developing strategies for disease management, potentially improving yields and mitigating losses. In mammals, interferon regulatory factors （IRFs） play a vital role in both the innate and adaptive immune system; though among teleosts IRF4 and IRFIO have seldom been studied. In this study, we characterized IRF4 and IRFIO from M. albus （malRF4 and malRFlO） and found that malRF4 cDNA consists of 1 716 nucleotides encoding a 451 amino acid （aa） protein, while malRFlO consists of 1 744 nucleotides including an open reading frame （ORF） of 1 236 nt encoding 411 aa. The malRFlO gene was constitutively expressed at high levels in a variety of tissues, while malRF4 showed a very limited expression pattern. Expression of malRF4 and malRFlO in head kidney, and spleen tissues was significantly up-regulated from 12 h to 48 h post-stimulation with polyinosinic： polycytidylic acid （poly I：C）, lipopolysaccharide （LPS） and a common pathogenic bacteria Aeromonas hydrophila. These results suggest that IRF4 and IRF10 play roles in immune responses to both viral and bacterial infections in M. albus.展开更多
Objective: Little is known about the cardiac contractility recovery after exercise. The objective of the study was to explore a method to evaluate the extent and speed of cardiac function up-regulation during exercise...Objective: Little is known about the cardiac contractility recovery after exercise. The objective of the study was to explore a method to evaluate the extent and speed of cardiac function up-regulation during exercise and the recovery course of cardiac contractility and heart rate after exercise. Methods: Ten student athletes and ten student non-athlete voluntarily participated in this controlled study. Three indicators were selected: 1) amplitude ratio of the first to second heart sound (S1/S2);2) heart rate (HR);3) power output (W). Phonocardiogram exercise test (PCGET) was adopted. A four-stage workload increment protocol was used. Phonocardiograms were recorded in the sitting position at rest and immediately after each test stage. The time taken for completing the workloads 1750 J, 3500 J, 5250 J, and 7000 J was recorded, respectively. During recovery heart sound signals were recorded immediately after exercise, and at 1, 5, 10, 15, 20, 25, and 30 minutes after exercise. S1/S2, HR, and W were calculated from the measured data. Cardiac function change trend graphs were constructed. Results: During exercise, HR and S1/S2 ratio increased with the increase in workload from 1750 J to 7000 J;the level and speed of increase in power output and S1/S2 ratio of the athletes were higher than the general students;power done by the general students decreased earlier than the athletes. During recovery course, the recovery course of the general students was slower than the athletes. Conclusion: This method for evaluating cardiac function up-regulation and recovery course is safe, easy, reliable, and effective, which is beneficial for selecting athletes, training, and matchmaking.展开更多
This study aimed to increase bacterial growth and 5-aminolevulinic acid（ALA） biosynthesis of Rhodobacter sphaeroides in wastewater treatment through adding ferrous ion（ Fe2＋ ）. Results demonstrated that Fe2＋ eff...This study aimed to increase bacterial growth and 5-aminolevulinic acid（ALA） biosynthesis of Rhodobacter sphaeroides in wastewater treatment through adding ferrous ion（ Fe2＋ ）. Results demonstrated that Fe2＋ effectively enhanced the biomass production and ALA yield of R. sphaeroides. Moreover, the optimal Fe2＋ dosage was found to be 400 μmol/L, which was associated with the highest biomass of 4015.3 mg/L and maximum ALA yield of 15.9 mg/g-dry cell weight（mg/g-DCW）. Mechanism analysis revealed that Fe2＋ vastly improved Adenosine Triphosphate（ATP） production by up-regulating the nif gene expression, and increasing ATP enhanced the biomass and ALA yield by supplying energy for bacterial growth and ALA biosynthesis, respectively. Correlation analysis showed that the ALA and ATP yields had positive relation with nifA and nifU gene expression. In addition, the nifA and nifU gene expression displayed high consistency of co-transcription at the optimal Fe2＋ dosage.展开更多
文摘AIM： To study the relationship between interleukin-lbeta （IL-1β） up-regulating tissue inhibitor of matrix metalloproteinase-1 （TIMMP-1） mRNA expression and phosphorylation of both c-jun N-terminal kinase （INK） and p38 in rat heffatic stellate cells （HSC）. METHODS： RT-PCR was performed to measure the expression of TIMMP-1 mRNA in rat HSC. Western blot was performed to measure IL-1β-induced JNK and p38 activities in rat HSC. RESULTS： TIMMP-1 mRNA expression （1.191± 0.079） was much higher after treatment with IL-1β （10 ng/mL） for 24 h than in control group （0.545±0.091） （P〈0.01）. IL-1β activated INK and p38 in a time-dependent manner. After stimulation with IL-1β for 0, 5, 15, 30, 60 and 120 min, the INK activity was 0.982±0.299, 1.501±0.720, 2.133±0.882, 3.360±0.452, 2.181±0.789, and 1.385 ± 0.368, respectively. There was a significant difference in JNK activity at 15 min （P〈 0.01）, 30 min （P〈 0.01） and 60 min （P〈0.01） in comparison to that at 0 min. The p38 activity was 1.061±0.310, 2.050±0.863, 2.380±0.573, 2.973±0.953, 2.421±0.793, and 1.755 ± 0.433 at the 6 time points （0, 5, 15, 30, 60 and 120 min） respectively. There was a significant difference in p38 activity at 5 min （P〈0.05）, 15 min （P〈0.01）, 30 min （P〈0.01） and 60 min （P〈0.01） compared to that at 0 min. TIMMP-1 mRNA expression trended to decrease in 3 groups pretreated with different concentrations of SP600125 （10 μmol/L, 1.022±0.113; 20 μmol/L, 0.869±0.070; 40 μmol/L, 0.666±0.123）. Their decreases were all significant （P〈0.05, P〈0.01, P〈0.01） in comparison to control group （without SP600125 treatment, 1.163±0.107）. In the other 3 groups pretreated with different concentrations of SB203580 （10 μmol/L, 1.507±0.099; 20 μmol/L, 1.698±0.107; 40 μmol/L, 1.857±0.054）, the expression of TIMMP-1 mRNA increased. Their levels were higher than those in the control group （without SB203580 treatment, 1.027 ± 0.061） with a si
基金supported by grants from the National Health and Medical Research Council of Australiathe Australian and New Zealand College of Anaesthetists
文摘Under normal conditions, the sympathetic neurotransmitter noradrenaline inhibits the production and release of pro-inflammatory cytokines. However, after peripheral nerve and tissue injury, pro-inflammatory cytokines appear to induce the expression of the alphalA-adreno- ceptor subtype on immune cells and perhaps also on other cells in the injured tissue. In turn, noradrenaline may act on up-regulated alphal-adrenoceptors to increase the production of the pro-inflammatory cytokine interleukin-6. In addition, the release of inflammatory mediators and nerve growth factor from keratinocytes and other cells may augment the expression of alphal-adrenoceptors on peripheral nerve fibers. Consequently, nociceptive afferents acquire an abnormal excitability to adrenergic agents, and inflammatory processes build. These mechanisms could contribute to the development of sympathetically maintained pain in conditions such as post-herpetic neuralgia, cutaneous neuromas, amputation stump pain and complex regional pain syndrome.
基金supported by the Shanghai Leading Academic Discipline Project (Grant No. B110)grants from CNHLPP (Grant No. 2005-BAC11A11)the Scientific Research Foundation for Young Talents of Shanghai Jiaotong University (Grant No. AB150200)
文摘Although prohibitin 1(PHB) is known to be associated with tumors,there are few studies regarding the role of PHB in hepatocellular carcinoma.An earlier glycoproteomics study of ours showed that PHB is over-expressed in MHCC97-H cells,a highly metastatic liver cancer cell line,and can be captured by wheat germ agglutinin.In the present study,western blotting and reverse transcription-PCR experiments showed an approximately 2-fold up-regulation in the expression of PHB in MHCC97-H cells.However,PHB was not significantly up-regulated in MHCC97-L cells,a low-metastatic liver cancer cell line.When PHB was over-expressed in MHCC97-L cells,cell proliferation was inhibited by 35% and migration increased about 2-fold.The results of this study show that PHB is up-regulated in MHCC97-H cells and is associated with both proliferation and migration of liver cancer cells.
基金Supported by National Natural Science Foundation of China,No.81170393
文摘AIM:To investigate cytokeratin 8(CK8)overexpression during hepatitis C virus(HCV)infection and its pathogenesis,and the effect of ectopic CK8 expression on hepatoma cell lines.METHODS:We successfully established an in vitro HCV cell culture system(HCVcc)to investigate the different expression profiles of CK8 in Huh-7-HCV and Huh-7.5-HCV cells.The expression of CK8 at the mRNA level was determined by real-time polymerase chain reaction(RT-PCR).The expression of CK8 at the protein level was evaluated by Western blotting.We then constructed a eukaryotic expression combination vector containing the coding sequence of human full length CK8 gene.CK8cDNA was amplified by reverse transcription-PCR and inserted into pEGFP-C1 and the positive clone pEGFPCK8 was obtained.After confirming the sequence,the recombinant plasmid was transfected into SMMC7721cells with lipofectamine2000 and CK8 expression was detected using inverted fluorescence microscopy,RTPCR and Western blotting.Besides,we identified biological function of CK8 on SMMC7721 cells,including cell proliferation,cell cycle and apoptosis detection.RESULTS:RT-PCR showed that the expression level of CK8 in Huh-7-HCV and Huh-7.5-HCV cells was 2.88and 2.95 times higher than in control cells.Western blot showed that CK8 expression in Huh-7-HCV and Huh-7.5-HCV cells was 2.53 and 3.26 times higher than that in control cells,respectively.We found that CK8at mRNA and protein levels were both significantly increased in HCVcc.CK8 was up-regulated in SMMC7721cells.CK8 expression at the mRNA level was significantly upregulated in SMMC7721/pEGFP-CK8 cells.CK8expression in SMMC7721/pEGFP-CK8 cells was 2.69times higher than in SMMC7721 cells,and was 2.64times higher than in SMMC7721/pEGFP-C1 cells.CK8expression at the protein level in SMMC7721/pEGFPCK8 cells was 2.46 times higher than in SMMC7721cells,and was 2.29 times higher than in SMMC7721/pEGFP-C1 cells.Further analysis demonstrated that forced expression of CK8 slowed cell growth and induced apoptosis of SMMC7721 cells.CONCLU
基金theMajorStateBasicResearchDevelopmentProgramofChina (No G19990 5 42 0 3)
文摘OBJECTIVE: To investigate the expression of Toll-like receptor 2 (TLR2) and Toll-like receptor 4 (TLR4) and the effect of lipopolysaccharide (LPS) on their expression in cultured endothelial cells. METHODS: Total RNA was extracted from ECV304 cells and isolated human umbilical vein endothelial cells (HUVECs) exposed to LPS, respectively. The quantification of TLR2 and TLR4 mRNA in HUVECs and EVC304 cells was carried out by reverse transcriptase-polymerase chain reaction (RT-PCR). RESULTS: ECV304 cells and HUVECs were able to express TLR2 and TLR4 mRNA, but the expression levels of TLR4 appeared to be stronger than those of TLR2. LPS could upregulate the expression levels of TLR4 obviously, whereas it had no effect on the expression level of TLR2. CONCLUSIONS: Our data indicate that TLR4 may be the LPS signal transducer in endothelial cells and plays important roles in the cell activation of LPS. The ECV304 cell line is a better experimental model than isolated HUVECs in the research of endothelial cells.
基金supported by grants from the National Natural Science Foundation of China (31571032, 31771191, and 81402903)supported by an Indiana Spinal Cord and Brain Injury Research Fund grant (ISCBIRF2017)
文摘Dear Editor,As the key contributor to the rising phase of action potentials in dorsal root ganglion（DRG）neurons,voltagegated sodium channels（VGSCs）are important in physiological and pathological pain conditions.For instance,abnormal expression of VGSCs in DRG neurons is the
基金financially supported by the Project from the National Natural Science Foundation of China(31101928)the State Key Laboratory of Freshwater Ecology and Biotechnology(2010FB02)Public Welfare Scientific Research Project of Hubei Province(2012DBA29001)
文摘The Asian swamp eel （Monopterus albus） is one of the most economically important freshwater fish in East Asia, but data on the immune genes of M. albus are scarce compared to other commercially important fish. A better understanding of the eel＇s immune responses may help in developing strategies for disease management, potentially improving yields and mitigating losses. In mammals, interferon regulatory factors （IRFs） play a vital role in both the innate and adaptive immune system; though among teleosts IRF4 and IRFIO have seldom been studied. In this study, we characterized IRF4 and IRFIO from M. albus （malRF4 and malRFlO） and found that malRF4 cDNA consists of 1 716 nucleotides encoding a 451 amino acid （aa） protein, while malRFlO consists of 1 744 nucleotides including an open reading frame （ORF） of 1 236 nt encoding 411 aa. The malRFlO gene was constitutively expressed at high levels in a variety of tissues, while malRF4 showed a very limited expression pattern. Expression of malRF4 and malRFlO in head kidney, and spleen tissues was significantly up-regulated from 12 h to 48 h post-stimulation with polyinosinic： polycytidylic acid （poly I：C）, lipopolysaccharide （LPS） and a common pathogenic bacteria Aeromonas hydrophila. These results suggest that IRF4 and IRF10 play roles in immune responses to both viral and bacterial infections in M. albus.
文摘Objective: Little is known about the cardiac contractility recovery after exercise. The objective of the study was to explore a method to evaluate the extent and speed of cardiac function up-regulation during exercise and the recovery course of cardiac contractility and heart rate after exercise. Methods: Ten student athletes and ten student non-athlete voluntarily participated in this controlled study. Three indicators were selected: 1) amplitude ratio of the first to second heart sound (S1/S2);2) heart rate (HR);3) power output (W). Phonocardiogram exercise test (PCGET) was adopted. A four-stage workload increment protocol was used. Phonocardiograms were recorded in the sitting position at rest and immediately after each test stage. The time taken for completing the workloads 1750 J, 3500 J, 5250 J, and 7000 J was recorded, respectively. During recovery heart sound signals were recorded immediately after exercise, and at 1, 5, 10, 15, 20, 25, and 30 minutes after exercise. S1/S2, HR, and W were calculated from the measured data. Cardiac function change trend graphs were constructed. Results: During exercise, HR and S1/S2 ratio increased with the increase in workload from 1750 J to 7000 J;the level and speed of increase in power output and S1/S2 ratio of the athletes were higher than the general students;power done by the general students decreased earlier than the athletes. During recovery course, the recovery course of the general students was slower than the athletes. Conclusion: This method for evaluating cardiac function up-regulation and recovery course is safe, easy, reliable, and effective, which is beneficial for selecting athletes, training, and matchmaking.
基金supported by the National Natural Science Foundation of China(No.51708214)the High-level Personnel Research Startup Project of North China University of Water Resources and Electric Power(No.40550)the Treatment Technology Integration and Demonstration for Domestic Sewage of Typical Villages and Towns in Henan Province(No.161100310700)
文摘This study aimed to increase bacterial growth and 5-aminolevulinic acid（ALA） biosynthesis of Rhodobacter sphaeroides in wastewater treatment through adding ferrous ion（ Fe2＋ ）. Results demonstrated that Fe2＋ effectively enhanced the biomass production and ALA yield of R. sphaeroides. Moreover, the optimal Fe2＋ dosage was found to be 400 μmol/L, which was associated with the highest biomass of 4015.3 mg/L and maximum ALA yield of 15.9 mg/g-dry cell weight（mg/g-DCW）. Mechanism analysis revealed that Fe2＋ vastly improved Adenosine Triphosphate（ATP） production by up-regulating the nif gene expression, and increasing ATP enhanced the biomass and ALA yield by supplying energy for bacterial growth and ALA biosynthesis, respectively. Correlation analysis showed that the ALA and ATP yields had positive relation with nifA and nifU gene expression. In addition, the nifA and nifU gene expression displayed high consistency of co-transcription at the optimal Fe2＋ dosage.
