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
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
In searching for differentially expressed genes in human uterine leiomyomas (ULs), suppression sub-tractive hybridization was used to construct an UL up-regulated library, which turned out to represent 88genes. After ...In searching for differentially expressed genes in human uterine leiomyomas (ULs), suppression sub-tractive hybridization was used to construct an UL up-regulated library, which turned out to represent 88genes. After two rounds of screening by reverse Northern analysis, twenty genes were proved to be up-regulated, including seventeen known genes and three genes with unknown function. All these genes werefirstly associated with UL. Three genes with notable difference were selected for Northern confirmationOur results proved the authenticity of the twenty genes. One gene named Phospholipase A2 (PLA2) showedup-regulation in 4/6 of the patients and investigation of tissue distribution indicated that it had obviousexpression in prostate, testis, liver, heart and skeletal muscle.展开更多
Monitoring expression at the transcriptional level is the first essential step for the functional analysis of plant genes. Genes-encoding proteins directly involved in early response to elicitor constitute only a smal...Monitoring expression at the transcriptional level is the first essential step for the functional analysis of plant genes. Genes-encoding proteins directly involved in early response to elicitor constitute only a small fraction of all the genes affected by elicitor. Transcriptional responses to various elicitors have been extensively studied in different plants including Nicotiana and Arabidopsis thaliana; however, corresponding data aren't available for non-heading Chinese cabbage. To address this problem, we describe a suppression subtractive library-based approach to isolate the plant's ESTs up-regulated in the early induction/execution of the HR induced by elicitor PB90 from Phytophthora boehmeriae. According to their putative identification in BLAST searches against the three genome databases, 70 up-regulated genes were classified into 9 parts: some aspect of primary 'metabolism' or 'energy' production; 'protein synthesis' or 'protein fate'; cellular communication/signal transduction mechanism; cell fates including Beclin, SPT1, and SPT2; HLA-B and AGO1 which participate in transcription; cellular transport and hypothetical proteins or proteins for which a function has yet to be determined. Seven selected genes such as Beclin, thioredoxin, HLA-B, MAP3K, SPT1, SPT2, and AGO1 were up-regulated induced by PB90, suggesting that the genes may play an important role in PB90-triggered HR.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.CONCLUSION:CK8 up-regulation might have a functional role in HCV infection and pathogenesis,and could be a promising target for the treatment of HCV infection.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
To explore the effect of environment conditions on immune activity of fish, eight immune-associated genes responsible for innate immunity were selected from the Gen Bank, i.e. Pgrn-a, Ifit2, P-hepcidin, Lect2, β2m, I...To explore the effect of environment conditions on immune activity of fish, eight immune-associated genes responsible for innate immunity were selected from the Gen Bank, i.e. Pgrn-a, Ifit2, P-hepcidin, Lect2, β2m, Irf1, Il25 and Hsp96, and the m RNA expressions of them in the kidney of cultured large yellow croaker Larimichthys crocea in different sea areas in the East China Sea were examined with q PCR techniques. In the contrasts of immune-associated gene expression between areas and populations, significant differences were found, expression levels of these immune-associated genes were lower in the clear water area than in the poor water quantity area, and lower in May than in October. MY was more sensitive to environmental factors than DQ, which was coincident with the water quality in the culturing areas. Differential analyses of the expression levels of these immune-associated genes showed that significant up-regulation could be triggered by poor environmental factors. The expression patterns indicated that the expression levels of these genes were sensitive to ecological changes, thereby the immune-associated genes, especially Pgrn-a, Ifit2, β2m, Il25 and Hsp96, might serve as immediate and sensitive indicators of population immunologic vigor and ecosystem health. But the expression of immunity-associated genes at the level of gene transcription is highly influenced by multiple factors, and the exact causes or influencing factors of the up-regulation or down-regulation of these genes still need further thorough investigation.展开更多
The regulatory mechanisms of drought and salt-associated miRNAs have not been fully understood in Sorghum bicolor. In this study, we investigated the effect of salinity stress (200 and 300 mM NaCl) and drought stress ...The regulatory mechanisms of drought and salt-associated miRNAs have not been fully understood in Sorghum bicolor. In this study, we investigated the effect of salinity stress (200 and 300 mM NaCl) and drought stress at pre- and post-flowering stages on the expression pattern of small regulatory RNAs in six Sorghum genotypes using semi-quantitative reverse transcriptase PCR (RT-qPCR). The results indicated that both drought and salt stresses altered the expression pattern of miRNAs in a dose-dependent manner. However, each miRNA responded to drought and salt stress in a different pattern among the six sorghum genotypes. miR156, miR167, miR168 and miR399 give different expressions levels compared to other studied miRNAs which may attribute to the adaption of sorghum to drought and salt stress and are good candidates for improving sorghum by transgenic technology.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
Insecticide resistance is an evolutionary adaptation that develops quite quickly in mosquitoes because of the high selection pressure of chemical insecticides, rapid generation time and large population size. Identifi...Insecticide resistance is an evolutionary adaptation that develops quite quickly in mosquitoes because of the high selection pressure of chemical insecticides, rapid generation time and large population size. Identification of genes associated with insecticide resistance is fundamental to understand the complex processes responsible for resistance. We compared the gene transcriptional profiles of parathion-resistant and -susceptible Culex pipiens quinquefasciatus using a combination of suppression subtractive hybridization and complementary DNA (cDNA) microarray techniques. A total of 278 colonies were selected from the resistant-susceptible mosquito subtractive library, 38 of which showed more than two fold stronger immunoblotting signals in the resistant strain than in the susceptible strain using cDNA microarray selection. The sequencing results showed that the 38 colonies can be matched to 12 genes of C. p. quinquefasciatus. Eight genes were confirmed to be overexpressed by more than two fold in the resistant strain. These genes encode chymotrypsin-1, theta glutathione S-transferase, lipase 3, larval serum protein 1β chain, cytochrome b, mitochondrial ribosomal large subunit, 28S rRNA, and a protein with unknown function. This study serves as a preliminary attempt to identify new genes associated with organophosphate resistance in this mosquito species and provides insights into the complicated physiological phenomenon of insecticide resistance.展开更多
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 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This work was supported by Nationa1 NaturalScience Fundation of China No.39700148 and LifeScience Special fund of CAS supported by ChineseMinisery of Finance.
文摘In searching for differentially expressed genes in human uterine leiomyomas (ULs), suppression sub-tractive hybridization was used to construct an UL up-regulated library, which turned out to represent 88genes. After two rounds of screening by reverse Northern analysis, twenty genes were proved to be up-regulated, including seventeen known genes and three genes with unknown function. All these genes werefirstly associated with UL. Three genes with notable difference were selected for Northern confirmationOur results proved the authenticity of the twenty genes. One gene named Phospholipase A2 (PLA2) showedup-regulation in 4/6 of the patients and investigation of tissue distribution indicated that it had obviousexpression in prostate, testis, liver, heart and skeletal muscle.
基金supported by the National Natural Science Foundation of China(30471123,30571206)Natural Science Foundation of Jiangsu Province,China(BK2005421)New Century Excellent Scholar Project of Ministry of Education of China(NCET-07-0042).
文摘Monitoring expression at the transcriptional level is the first essential step for the functional analysis of plant genes. Genes-encoding proteins directly involved in early response to elicitor constitute only a small fraction of all the genes affected by elicitor. Transcriptional responses to various elicitors have been extensively studied in different plants including Nicotiana and Arabidopsis thaliana; however, corresponding data aren't available for non-heading Chinese cabbage. To address this problem, we describe a suppression subtractive library-based approach to isolate the plant's ESTs up-regulated in the early induction/execution of the HR induced by elicitor PB90 from Phytophthora boehmeriae. According to their putative identification in BLAST searches against the three genome databases, 70 up-regulated genes were classified into 9 parts: some aspect of primary 'metabolism' or 'energy' production; 'protein synthesis' or 'protein fate'; cellular communication/signal transduction mechanism; cell fates including Beclin, SPT1, and SPT2; HLA-B and AGO1 which participate in transcription; cellular transport and hypothetical proteins or proteins for which a function has yet to be determined. Seven selected genes such as Beclin, thioredoxin, HLA-B, MAP3K, SPT1, SPT2, and AGO1 were up-regulated induced by PB90, suggesting that the genes may play an important role in PB90-triggered HR.
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.CONCLUSION:CK8 up-regulation might have a functional role in HCV infection and pathogenesis,and could be a promising target for the treatment of HCV infection.
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provided by The National High Technology Research and Development Program of China (863 Program) (No.2012BAB16B02)
文摘To explore the effect of environment conditions on immune activity of fish, eight immune-associated genes responsible for innate immunity were selected from the Gen Bank, i.e. Pgrn-a, Ifit2, P-hepcidin, Lect2, β2m, Irf1, Il25 and Hsp96, and the m RNA expressions of them in the kidney of cultured large yellow croaker Larimichthys crocea in different sea areas in the East China Sea were examined with q PCR techniques. In the contrasts of immune-associated gene expression between areas and populations, significant differences were found, expression levels of these immune-associated genes were lower in the clear water area than in the poor water quantity area, and lower in May than in October. MY was more sensitive to environmental factors than DQ, which was coincident with the water quality in the culturing areas. Differential analyses of the expression levels of these immune-associated genes showed that significant up-regulation could be triggered by poor environmental factors. The expression patterns indicated that the expression levels of these genes were sensitive to ecological changes, thereby the immune-associated genes, especially Pgrn-a, Ifit2, β2m, Il25 and Hsp96, might serve as immediate and sensitive indicators of population immunologic vigor and ecosystem health. But the expression of immunity-associated genes at the level of gene transcription is highly influenced by multiple factors, and the exact causes or influencing factors of the up-regulation or down-regulation of these genes still need further thorough investigation.
文摘The regulatory mechanisms of drought and salt-associated miRNAs have not been fully understood in Sorghum bicolor. In this study, we investigated the effect of salinity stress (200 and 300 mM NaCl) and drought stress at pre- and post-flowering stages on the expression pattern of small regulatory RNAs in six Sorghum genotypes using semi-quantitative reverse transcriptase PCR (RT-qPCR). The results indicated that both drought and salt stresses altered the expression pattern of miRNAs in a dose-dependent manner. However, each miRNA responded to drought and salt stress in a different pattern among the six sorghum genotypes. miR156, miR167, miR168 and miR399 give different expressions levels compared to other studied miRNAs which may attribute to the adaption of sorghum to drought and salt stress and are good candidates for improving sorghum by transgenic technology.
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.
基金Acknowledgments This study was financially supported by the National Natural Science Foundation of China (Grant No. 31272364) and the Important National Science & Technology Specific Project of China (Grant No. 2012ZX 10004219).
文摘Insecticide resistance is an evolutionary adaptation that develops quite quickly in mosquitoes because of the high selection pressure of chemical insecticides, rapid generation time and large population size. Identification of genes associated with insecticide resistance is fundamental to understand the complex processes responsible for resistance. We compared the gene transcriptional profiles of parathion-resistant and -susceptible Culex pipiens quinquefasciatus using a combination of suppression subtractive hybridization and complementary DNA (cDNA) microarray techniques. A total of 278 colonies were selected from the resistant-susceptible mosquito subtractive library, 38 of which showed more than two fold stronger immunoblotting signals in the resistant strain than in the susceptible strain using cDNA microarray selection. The sequencing results showed that the 38 colonies can be matched to 12 genes of C. p. quinquefasciatus. Eight genes were confirmed to be overexpressed by more than two fold in the resistant strain. These genes encode chymotrypsin-1, theta glutathione S-transferase, lipase 3, larval serum protein 1β chain, cytochrome b, mitochondrial ribosomal large subunit, 28S rRNA, and a protein with unknown function. This study serves as a preliminary attempt to identify new genes associated with organophosphate resistance in this mosquito species and provides insights into the complicated physiological phenomenon of insecticide resistance.