[ Objective ] The aim of the research was to study the expression profile changes of genes involved in lipid metabolism pathway during liver regeneration in mice. [ Method] The CCI4 induced mouse model of liver regene...[ Objective ] The aim of the research was to study the expression profile changes of genes involved in lipid metabolism pathway during liver regeneration in mice. [ Method] The CCI4 induced mouse model of liver regeneration was established and the total RNA was isolated from liver tissue of mouse. Then the changes of genes involved in lipid metabolism pathway during different stages of liver regeneration were detected through micro-array chip gene technique and their specific functions were also analyzed. [ Result] Dudng the process of liver regeneration, the expression level of 98 genes involved in lipid metabolism pathway changed, which were divided into eight groups according to change trend. In the mass, the expression of genes was inhibited in the early stage and up-regulated in the late phase. And the gene expression associated with fatty acid synthesis pathway was mainly up-regulated while the catabolic pathway did not change significantly. Most of genes involved in bile acid synthesis pathway were suppressed before 4.5 d and up-regulated after 4.5 d or 7 d. [ Conclusion] During the process of liver regeneration, the genes associated with lipid metabolism are expressed in different trends, and this data should provide a specific range of genes for further studying the regulation effect of lipid metabolism related pathway on liver regeneration.展开更多
The ripening process of grape is an important stage during grape growth and development. During this process, color of grape skin is the most obvious change. The molecular mechanism for the ripening of grape(a non-cli...The ripening process of grape is an important stage during grape growth and development. During this process, color of grape skin is the most obvious change. The molecular mechanism for the ripening of grape(a non-climacteric fruit, which ripens without ethylene and respiration bursts) is still unclear. Although numerous studies have been done on the changes in the contents of metabolites during grape ripening, the differentially expressed genes at veraison and maturity stages have not been systematically analyzed. In this study, 1 524 genes that are significantly differentially expressed in grape(Pinot Noir) skin at veraison and maturity stages were identified, and a co-expression network of these genes was built. Some of the eight co-expression modules we identified may be closely related to the synthesis or metabolism of anthocyanins, sugar acids, and other flavor substances. The transcription factor families WRKY, b ZIP, HSF and WOX may play an important role in the regulation of anthocyanin synthesis or metabolism. The results provide a foundation for further study of the molecular mechanism of grape ripening.展开更多
Pepsinogens are zymogens of pepsins,aspecific proteases working as digestive enzymes in vertebrate stomach,of which biological and molecular properties have been extensively studied.Several exhaustive studies have bee...Pepsinogens are zymogens of pepsins,aspecific proteases working as digestive enzymes in vertebrate stomach,of which biological and molecular properties have been extensively studied.Several exhaustive studies have been performed in the pepsinogen producing cells in developing rat stomachs,but little is known about the expression of pepsinogen gene in these cells.In this study,the ontogeny of pepsinogen producing cells in rat fundic glands was studied by in situ hybridization using a digoxigenin-labeled RNA probe.The rat gastric epithelium was stratified but was morphologically undifferentiated at the stage of 18.5 days of gestation.The pepsinogen mRNA was expressed both in chief cells and mucous neck cells in adult rats,which was first detected by in situ hybridization in the stomach of the rats at 3.5 days after birth.The development of pepsinogen producing cells could be classified into four stages:(1) 18.5 days of gestation to 0.5 day after birth;(2) 3.5 days to 2 weeks after birth;(3) 3～4 weeks after birth;(4) 8 weeks after birth.Pepsinogen expression is strictly limited to these cells,the distribution of which shown a developmental stage-specific manner.We concluded the pepsinogen C could offer excellent molecular markers of differentiation during stomach epithelial cellulur development.展开更多
AIM:To investigate the effect of in vivo environment on gene expression in Helicobacter pylori(H.pylori) as it relates to its survival in the host.METHODS:In vivo expression technology(IVET) systems are used to identi...AIM:To investigate the effect of in vivo environment on gene expression in Helicobacter pylori(H.pylori) as it relates to its survival in the host.METHODS:In vivo expression technology(IVET) systems are used to identify microbial virulence genes.We modified the IVET-transcriptional fusion vector,pIVET8,which uses antibiotic resistance as the basis for selection of candidate genes in host tissues to develop two unique IVET-promoter-screening vectors,pIVET11 and pIVET12.Our novel IVET systems were developed by the fusion of random Sau3A DNA fragments of H.pylori and a tandem-reporter system of chloramphenicol acetyltransferase and beta-galactosidase.Additionally,each vector contains a kanamycin resistance gene.We used a mouse macrophage cell line,RAW 264.7 and mice,as selective media to identify specific genes that H.pylori expresses in vivo.Gene expression studies were conducted by infecting RAW 264.7 cells with H.pylori.This was followed by real time polymerase chain reaction(PCR) analysis to determine the relative expression levels of in vivo induced genes.RESULTS:In this study,we have identified 31 in vivo induced(ivi) genes in the initial screens.These 31 genes belong to several functional gene families,including several well-known virulence factors that are expressed by the bacterium in infected mouse stomachs.Virulence factors,vacA and cagA,were found in this screen and are known to play important roles in H.pylori infection,colonization and pathogenesis.Their detection validates the efficacy of these screening systems.Some of the identified ivi genes have already been implicated to play an important role in the pathogenesis of H.pylori and other bacterial pathogens such as Escherichia coli and Vibrio cholerae.Transcription profiles of allivi genes were confirmed by real time PCR analysis of H.pylori RNA isolated from H.pylori infected RAW 264.7 macrophages.We compared the expression profile of H.pylori and RAW 264.7 coculture with that of H.pylori only.Some genes such as cag A,vac A,lpx C,mur I,tlp C,trx B,sod B,tnp B,pgi,rbf A and inf B showed a 2-20 fold upregulation.Statistically significant upregulation was obtained for all the above mentioned genes(P < 0.05).tlp C,cag A,vac A,sod B,rbf A,inf B,tnp B,lpx C and mur I were also significantly upregulated(P < 0.01).These data suggest a strong correlation between results obtained in vitro in the macrophage cell line and in the intact animal.CONCLUSION:The positive identification of these genes demonstrates that our IVET systems are powerful tools for studying H.pylori gene expression in the host environment.展开更多
3,4-Dihydroxy-2-butanone 4-phosphate （DHBP） and GTP are the precursors for riboflavin biosynthesis. In this research, improving the precursor supply for riboflavin production was attempted by overexpressing ribB and...3,4-Dihydroxy-2-butanone 4-phosphate （DHBP） and GTP are the precursors for riboflavin biosynthesis. In this research, improving the precursor supply for riboflavin production was attempted by overexpressing ribB and engineering purine pathway in a riboflavin-producing Escherichia colt strain. Initially, ribB gene was overexpressed to increase the flux from ribulose 5-phosphate （Ru-5-P） to DHBP. Then ndk and grnk genes were overexpressed to enhance GTP supply. Subsequently, a R419L mutation was introduced into purA to reduce the flux from IMP to AMP. Finally, co-overexpression of mutant purF and prs genes further increased riboflavin production. The final strain RF18S produced 387.6 mg riboflavin · L-1 with a yield of 44.8 mg riboflavin per gram glucose in shake-flask fermentations. The final titer and yield were 72.2% and 55.6% higher than those of RF01S, respectively. It was concluded that simultaneously engineering the DHBP synthase and GTP biosynthetic pathway by rational metabolic engineering can efficiently boost riboflavin production in E. coll.展开更多
In this study, we examined the effect of elevated temperature on the expression patterns of genes, i.e., nacrein, irr, n16, n19, and hsp70 in the pearl oyster Pinctada fucata. The experiment was carried out at 4 tempe...In this study, we examined the effect of elevated temperature on the expression patterns of genes, i.e., nacrein, irr, n16, n19, and hsp70 in the pearl oyster Pinctada fucata. The experiment was carried out at 4 temperatures, i.e., 20℃(ambient, control), 24, 28℃, and 32℃. The expression levels of target genes in P. fucata were assayed at 0, 6, 24, 48, and 96 h via real-time polymerase chain reaction. Results showed that the expression levels of nacrein and irr had no significant variations among different time points below 28℃, but significantly increased over time at 32℃. The expression levels of n16 and n19 did not change markedly at 20℃. The former increased significantly at 6 h and 24 h while the latter substantially decreased during 6–96 h at 24, 28 and 32℃. Among different temperatures, the level of n16 was significantly lower at 20℃ than at other temperatures during 6–96 h, and the level of n19 significantly varied among different temperatures at 48 h and 96 h. The expression level of hsp70 was significantly higher at 32℃ than at 20, 24 and 28℃ at 24 h. These results demonstrated that elevated temperature impacted the physiological processes of P. fucata and potentially influenced its adaptability to thermal stress.展开更多
Objective： To evaluate the relationship between the expressions of cyclin D1 and p27^kip1 in the canceration course of the stomach. Methods： The immunohistochemical staining technique （SP method） was used to detec...Objective： To evaluate the relationship between the expressions of cyclin D1 and p27^kip1 in the canceration course of the stomach. Methods： The immunohistochemical staining technique （SP method） was used to detect the expressions of cyclin D1, p27^kip1 in chronic superficial gastritis （CSG）, chronic atrophic gastritis （CAG）, intestinal metaplasia （IM）, dysplasia （DYS）, gastric carcinoma （GCA） biopsy specimens. Results： The positive cyclin D1 expression rates increased with the progressing from CAG--,IM--,DYS--,GCA respectively, and those in IM, DYS and GCA were different from those in CSG, P 〈 0.05, while DYS group was indifferent from GCA group, P 〉 0.05. The positive p27^kip1 expression rates decreased with the mucosa progressing from CAG→IM→DYS→GCA. There was a negative correlation between the expression cyclin D1 and p27^kip1 （y = -0.53, P = 0.000）. Conclusion： Expression rates of cyclin D1 in the canceration course of the stomach mucosa trend were increased and those of p27^kip1 were decreased; the abnormal expressions of them were found in the early term of the canceration course of the stomach mucosa, and the inverse expression suggests there may be a negative feedback regulatory loop between cyclin D1 and p27^kip1.展开更多
文摘[ Objective ] The aim of the research was to study the expression profile changes of genes involved in lipid metabolism pathway during liver regeneration in mice. [ Method] The CCI4 induced mouse model of liver regeneration was established and the total RNA was isolated from liver tissue of mouse. Then the changes of genes involved in lipid metabolism pathway during different stages of liver regeneration were detected through micro-array chip gene technique and their specific functions were also analyzed. [ Result] Dudng the process of liver regeneration, the expression level of 98 genes involved in lipid metabolism pathway changed, which were divided into eight groups according to change trend. In the mass, the expression of genes was inhibited in the early stage and up-regulated in the late phase. And the gene expression associated with fatty acid synthesis pathway was mainly up-regulated while the catabolic pathway did not change significantly. Most of genes involved in bile acid synthesis pathway were suppressed before 4.5 d and up-regulated after 4.5 d or 7 d. [ Conclusion] During the process of liver regeneration, the genes associated with lipid metabolism are expressed in different trends, and this data should provide a specific range of genes for further studying the regulation effect of lipid metabolism related pathway on liver regeneration.
基金Supported by Major Agricultural Application Technology Innovation Project of Shandong Province"Development of Landmark Wines and Integrated Application of Key Technologies in Shandong Province"Major Agricultural Application Technology Innovation Project of Shandong Province"Research and Application of Precision Control of Maturation and Product Innovation of Featured Brewing Grape"Agricultural Scientific and Technological Innovation Project of Shandong Academy of Agricultural Sciences(CXGC2016D01)
文摘The ripening process of grape is an important stage during grape growth and development. During this process, color of grape skin is the most obvious change. The molecular mechanism for the ripening of grape(a non-climacteric fruit, which ripens without ethylene and respiration bursts) is still unclear. Although numerous studies have been done on the changes in the contents of metabolites during grape ripening, the differentially expressed genes at veraison and maturity stages have not been systematically analyzed. In this study, 1 524 genes that are significantly differentially expressed in grape(Pinot Noir) skin at veraison and maturity stages were identified, and a co-expression network of these genes was built. Some of the eight co-expression modules we identified may be closely related to the synthesis or metabolism of anthocyanins, sugar acids, and other flavor substances. The transcription factor families WRKY, b ZIP, HSF and WOX may play an important role in the regulation of anthocyanin synthesis or metabolism. The results provide a foundation for further study of the molecular mechanism of grape ripening.
文摘Pepsinogens are zymogens of pepsins,aspecific proteases working as digestive enzymes in vertebrate stomach,of which biological and molecular properties have been extensively studied.Several exhaustive studies have been performed in the pepsinogen producing cells in developing rat stomachs,but little is known about the expression of pepsinogen gene in these cells.In this study,the ontogeny of pepsinogen producing cells in rat fundic glands was studied by in situ hybridization using a digoxigenin-labeled RNA probe.The rat gastric epithelium was stratified but was morphologically undifferentiated at the stage of 18.5 days of gestation.The pepsinogen mRNA was expressed both in chief cells and mucous neck cells in adult rats,which was first detected by in situ hybridization in the stomach of the rats at 3.5 days after birth.The development of pepsinogen producing cells could be classified into four stages:(1) 18.5 days of gestation to 0.5 day after birth;(2) 3.5 days to 2 weeks after birth;(3) 3～4 weeks after birth;(4) 8 weeks after birth.Pepsinogen expression is strictly limited to these cells,the distribution of which shown a developmental stage-specific manner.We concluded the pepsinogen C could offer excellent molecular markers of differentiation during stomach epithelial cellulur development.
基金Supported by Intramural Research Program of the National Institutes of Health,National Institute of Diabetes and Digestive and Kidney DiseaseThe Division of Intramural Research of the National Institute of Allergy and Infectious DiseasesAn Inter-Agency Agreement (Y3-DK-3521-07) with the National Institute on Minority Health and Health Disparities
文摘AIM:To investigate the effect of in vivo environment on gene expression in Helicobacter pylori(H.pylori) as it relates to its survival in the host.METHODS:In vivo expression technology(IVET) systems are used to identify microbial virulence genes.We modified the IVET-transcriptional fusion vector,pIVET8,which uses antibiotic resistance as the basis for selection of candidate genes in host tissues to develop two unique IVET-promoter-screening vectors,pIVET11 and pIVET12.Our novel IVET systems were developed by the fusion of random Sau3A DNA fragments of H.pylori and a tandem-reporter system of chloramphenicol acetyltransferase and beta-galactosidase.Additionally,each vector contains a kanamycin resistance gene.We used a mouse macrophage cell line,RAW 264.7 and mice,as selective media to identify specific genes that H.pylori expresses in vivo.Gene expression studies were conducted by infecting RAW 264.7 cells with H.pylori.This was followed by real time polymerase chain reaction(PCR) analysis to determine the relative expression levels of in vivo induced genes.RESULTS:In this study,we have identified 31 in vivo induced(ivi) genes in the initial screens.These 31 genes belong to several functional gene families,including several well-known virulence factors that are expressed by the bacterium in infected mouse stomachs.Virulence factors,vacA and cagA,were found in this screen and are known to play important roles in H.pylori infection,colonization and pathogenesis.Their detection validates the efficacy of these screening systems.Some of the identified ivi genes have already been implicated to play an important role in the pathogenesis of H.pylori and other bacterial pathogens such as Escherichia coli and Vibrio cholerae.Transcription profiles of allivi genes were confirmed by real time PCR analysis of H.pylori RNA isolated from H.pylori infected RAW 264.7 macrophages.We compared the expression profile of H.pylori and RAW 264.7 coculture with that of H.pylori only.Some genes such as cag A,vac A,lpx C,mur I,tlp C,trx B,sod B,tnp B,pgi,rbf A and inf B showed a 2-20 fold upregulation.Statistically significant upregulation was obtained for all the above mentioned genes(P < 0.05).tlp C,cag A,vac A,sod B,rbf A,inf B,tnp B,lpx C and mur I were also significantly upregulated(P < 0.01).These data suggest a strong correlation between results obtained in vitro in the macrophage cell line and in the intact animal.CONCLUSION:The positive identification of these genes demonstrates that our IVET systems are powerful tools for studying H.pylori gene expression in the host environment.
基金supported by National High-tech R&D Program of China [2012AA02A702, 2012AA022103]
文摘3,4-Dihydroxy-2-butanone 4-phosphate （DHBP） and GTP are the precursors for riboflavin biosynthesis. In this research, improving the precursor supply for riboflavin production was attempted by overexpressing ribB and engineering purine pathway in a riboflavin-producing Escherichia colt strain. Initially, ribB gene was overexpressed to increase the flux from ribulose 5-phosphate （Ru-5-P） to DHBP. Then ndk and grnk genes were overexpressed to enhance GTP supply. Subsequently, a R419L mutation was introduced into purA to reduce the flux from IMP to AMP. Finally, co-overexpression of mutant purF and prs genes further increased riboflavin production. The final strain RF18S produced 387.6 mg riboflavin · L-1 with a yield of 44.8 mg riboflavin per gram glucose in shake-flask fermentations. The final titer and yield were 72.2% and 55.6% higher than those of RF01S, respectively. It was concluded that simultaneously engineering the DHBP synthase and GTP biosynthetic pathway by rational metabolic engineering can efficiently boost riboflavin production in E. coll.
基金supported by the National Natural Science Foundation of China (41006090)Joint Program of NSFC-Guangdong (U0831001)the Funds of Knowledge Innovation Program of Chinese Academy of Sciences (ZCX2-EW-Q21)
文摘In this study, we examined the effect of elevated temperature on the expression patterns of genes, i.e., nacrein, irr, n16, n19, and hsp70 in the pearl oyster Pinctada fucata. The experiment was carried out at 4 temperatures, i.e., 20℃(ambient, control), 24, 28℃, and 32℃. The expression levels of target genes in P. fucata were assayed at 0, 6, 24, 48, and 96 h via real-time polymerase chain reaction. Results showed that the expression levels of nacrein and irr had no significant variations among different time points below 28℃, but significantly increased over time at 32℃. The expression levels of n16 and n19 did not change markedly at 20℃. The former increased significantly at 6 h and 24 h while the latter substantially decreased during 6–96 h at 24, 28 and 32℃. Among different temperatures, the level of n16 was significantly lower at 20℃ than at other temperatures during 6–96 h, and the level of n19 significantly varied among different temperatures at 48 h and 96 h. The expression level of hsp70 was significantly higher at 32℃ than at 20, 24 and 28℃ at 24 h. These results demonstrated that elevated temperature impacted the physiological processes of P. fucata and potentially influenced its adaptability to thermal stress.
文摘Objective： To evaluate the relationship between the expressions of cyclin D1 and p27^kip1 in the canceration course of the stomach. Methods： The immunohistochemical staining technique （SP method） was used to detect the expressions of cyclin D1, p27^kip1 in chronic superficial gastritis （CSG）, chronic atrophic gastritis （CAG）, intestinal metaplasia （IM）, dysplasia （DYS）, gastric carcinoma （GCA） biopsy specimens. Results： The positive cyclin D1 expression rates increased with the progressing from CAG--,IM--,DYS--,GCA respectively, and those in IM, DYS and GCA were different from those in CSG, P 〈 0.05, while DYS group was indifferent from GCA group, P 〉 0.05. The positive p27^kip1 expression rates decreased with the mucosa progressing from CAG→IM→DYS→GCA. There was a negative correlation between the expression cyclin D1 and p27^kip1 （y = -0.53, P = 0.000）. Conclusion： Expression rates of cyclin D1 in the canceration course of the stomach mucosa trend were increased and those of p27^kip1 were decreased; the abnormal expressions of them were found in the early term of the canceration course of the stomach mucosa, and the inverse expression suggests there may be a negative feedback regulatory loop between cyclin D1 and p27^kip1.
