[Objective] The purpose of this study was to construct a vscC in-frame deletion mutant of Vibrio alginolyticus with no antibiotic resistance marker. [Method] The first vscC mutant molecules in vitro were generated by ...[Objective] The purpose of this study was to construct a vscC in-frame deletion mutant of Vibrio alginolyticus with no antibiotic resistance marker. [Method] The first vscC mutant molecules in vitro were generated by SOE-PCR and then lig- ated to a suicide vector pDM4 to construct a suicide recombinant vector pDM4- A vscC. To clone the recombinant vector, it was transformed to E. coli SY327 strain, and then positive clones were selected and proved by PCR analysis. After that, the pDM4-AvscC DNA was extracted in large numbers and transformed to the E. coil S17-1 strain that acted as a donor in bacterial conjugation using the heat shock method. The recombinant E. coli S17-1 strains then transferred the pDM4-AvscC to V. alginolyticus ZJ51-O by conjugation method; transconjugants were screened and selected sequentially using antibiotic selection strategy and sucrose based counter-selection system to find the suspected mutants wanted. Finally the putative mutants were identified by PCR and confirmed by sequencing analysis. [Result] ZJ51-OAvscC was successfully constructed. [Conclusion] This study laid foundation for further research on the function of vscC gene and molecular mechanism of type Ⅲ secretion system in V. alginolyticus. Simultaneously, by the effective method other unknown functional genes in V. alginolyticus genome would be researched.展开更多
The presence of DNA homologues corresponding to verc203 (vernalization-related cDNA clone) was investigated by molecular hybridization techniques. The genes were detected in 16 plant species that cover 12 subclasses o...The presence of DNA homologues corresponding to verc203 (vernalization-related cDNA clone) was investigated by molecular hybridization techniques. The genes were detected in 16 plant species that cover 12 subclasses of the Takhtajan system of angiosperms classification including diverse model species. The results of Southern blot analysis showed a low copy number of this gene existed in rice, wheat, barley and Arabidopsis. The hybridization result of PCR products demonstrated the conservation of the gene corresponding to ver203 in diverse plants. The phylogenetic tree of the ver203 gene in tested plants was supported by evolution relationship of species. The ver203 gene expressed in a vernalized plumule winter wheat, instead of the root. And the endosperm before the treatment was essential for the ver203 expression during vernalization in wheat. In Arabidopsis thaliana, the pattern of expression showed that the gene corresponding to ver203 was expressed at low temperature for 14 days. Gibberellin(GA3) may accelerate the expression of ver203 gene in Arabidopsis exposed to low temperature. However, it could not replace vernalization treatment to initiate the gene expression.展开更多
Matrix effects can significantly hamper the accuracy and precision of the analysis results of perfluorinated acids (PFAs) in environmental solid samples. Several methods, such as standard addition, isotopically labe...Matrix effects can significantly hamper the accuracy and precision of the analysis results of perfluorinated acids (PFAs) in environmental solid samples. Several methods, such as standard addition, isotopically labeled internal standards, clean-up of SPE (solid phase extraction) eluents by dispersive graphitized carbon sorbent and substitution of eletrospray ionization (ESI) source by atmosphere pressure photoionization (APPI) source, were demonstrated for elimination of matrix effects in quantitative analysis of PFAs in solid samples. The resuRs indicate that matrix effects can be effectively eliminated by standard addition, but instrumental analysis time will be multiplied. Isotopically labeled internal standards can effectively negate matrix effects of PFAs with the same perfluorocarbon chain length, but is not valid for the other analytes. Although APPI can eliminate matrix effects for all analytes, it is only suitable for analysis of high pollution levels samples. Clean-up of SPE eluents by dispersive graphitized carbon sorbent not only effectively negate the impact of matrix effect, but also avoid frequent clean of the ESI in order to maintain instrumental sensitivity. Therefore, the best method for elimination of matrix effects is the usage of dispersive graphitized carbon sorbent for clean-up of SPE elution.展开更多
AIM:To develop lymph node metastasis(LNM)-associated biomarkers for colorectal cancer(CRC) using quantitative proteome analysis.METHODS:Differences in protein expression between primary CRC with LNM(LNM CRC) and witho...AIM:To develop lymph node metastasis(LNM)-associated biomarkers for colorectal cancer(CRC) using quantitative proteome analysis.METHODS:Differences in protein expression between primary CRC with LNM(LNM CRC) and without LNM(non-LNM CRC) were assessed using methyl esterification stable isotope labeling coupled with 2D liquid chromatography followed by tandem mass spectrometry(2D-LC-MS/MS).The relationship to clinicopathological parameters and prognosis of candidate biomarkers was examined using an independent sample set.RESULTS:Forty-three proteins were found to be differentially expressed by at least 2.5-fold in two types of CRC.S100A4 was significantly upregulated in LNM CRC compared with non-LNM CRC,which was confirmed by Western blotting,immunohistochemistry and real-time quantitative polymerase chain reaction.Further immunohistochemistry on another 112 CRC cases showed that overexpression of S100A4 frequently existed in LNM CRC compared with non-LNM CRC(P < 0.001).Overexpression of S100A4 was significantly associated with LNM(P < 0.001),advanced TNM stage(P < 0.001),increased 5-year recurrence rate(P < 0.001) and decreased 5-year overall survival rate(P < 0.001).Univariate and multivariate analyses indicated that S100A4 expression was an independent prognostic factor for recurrence and survival of CRC patients(P < 0.05).CONCLUSION:S100A4 might serve as a powerful biomarker for LNM and a prognostic factor in CRC.展开更多
Objective : To evaluate the changes of serum matrix metalloproteinase-9 (romp-9) in patients of incipient diabetic nephropathy with or without macrovascular disease and to analyze the factors associated with homocy...Objective : To evaluate the changes of serum matrix metalloproteinase-9 (romp-9) in patients of incipient diabetic nephropathy with or without macrovascular disease and to analyze the factors associated with homocysteine(hcy), interleukin-6(IL-6), tumor necrosis factor-alpha (TNF-α), highly sensitive C-reactive protein (hsCRP), HbAlc and lipid profile in those patients in order to know whether this marker or other factors are more important to induce diabetic macrovascular disease. Methods: Type 2 diabetes mellitus(T2DM) subjects with incipient diabetic nephropathy with or without macrovascular disease were selected for participation and divided into 2 groups. The patients in group 1 (n= 38) used insulin, and patients in group 2 (n=34) were treated with an oral antidiabetic drug. Then serum mmp-9, hey, IL-6 and TNF-α in these patients were measured, and compared to the healthy subjects as control (n= 16). The results were analyzed by SPSS13. Results: Serum romp-9 and hcy of the patients having incipient diabetic nephropathy with macrovascular disease were higher than that of patients without macrovascular disease (P〈0.01). For insulin-injected patients, whether they accompanied with macrovascular diseases or not, the serum levels of romp-9, hcy, IL-6 and TNF-α were all lower, but no significant statistics compared with non-insulin used patients or the healthy subjects. The serum level of romp-9 was more correlated with the serum hcy in antidiabetic drug used patients. (P〈0. 000) Conclusion: The serum level of romp-9 plays an important role of pathogenesis in the macrovascular disease in the incipient diabetic patients, and the serum level of hcy also can reflect the severely degree of macrovascular disease in these patients, insulin can reduce these markers.展开更多
Global climate change poses a severe threat to mountain biodiversity.Phenotypic plasticity and local adaptation are two common strategies for alpine plant to cope with such change.They may facilitate organismal adapta...Global climate change poses a severe threat to mountain biodiversity.Phenotypic plasticity and local adaptation are two common strategies for alpine plant to cope with such change.They may facilitate organismal adaptation to contrasting environments,depending on the influences of the environment or genotype or their interacted effects.In this study,we use an endemic alpine plant(Rorippa elata)in the Hengduan mountains(HDM)to unravel its phenotypic basis of adaptation strategy and evaluate the relative contributions of environment and genotype to its phenotype.We transplanted 37 genotypes of R.elata into two common gardens across low and high elevations(2800 vs.3800 m)during 2021-2022.Nine fitness-related traits were measured,including flowering probability and glucosinolates(GS)content.We estimated the environmental or genotypic contributions to the phenotype and identified the main environmental components.Our results revealed that both environment and genotype-by-environment interactions contributed to the phenotypes of R.elata.Latitudinal heterogeneity was identified as a key factor that explained 24%of the total phenotypic variation.In particular,genotypes of the northern HDM showed significantly higher plasticity in flowering probability than those of the southern HDM.Furthermore,within the southern HDM,GS content indicated local adaptation to herbivory stresses for R.elata genotypes along elevations.In conclusion,our results suggest that R.elata may have adapted to the alpine environment through species-level plasticity or regional-level local adaptation.These processes were shaped by either complex topography or interactions between genotype and mountain environments.Our study provides empirical evidence on the adaptation of alpine plants.展开更多
Long-term meteorological observation series are fundamental for reflecting climate changes.However,almost all meteorological stations inevitably undergo relocation or changes in observation instruments,rules,and metho...Long-term meteorological observation series are fundamental for reflecting climate changes.However,almost all meteorological stations inevitably undergo relocation or changes in observation instruments,rules,and methods,which can result in systematic biases in the observation series for corresponding periods.Homogenization is a technique for adjusting these biases in order to assess the true trends in the time series.In recent years,homogenization has shifted its focus from the adjustments to climate mean status to the adjustments to information about climate extremes or extreme weather.Using case analyses of ideal and actual climate series,here we demonstrate the basic idea of homogenization,introduce new understanding obtained from recent studies of homogenization of climate series in China,and raise issues for further studies in this field,especially with regards to climate extremes,uncertainty of the statistical adjustments,and biased physical relationships among different climate variables due to adjustments in single variable series.展开更多
The ground state properties of Ds(Z=110) isotopes(N=151-195) are studied in the framework of the relativistic mean field(RMF) theory with the effective interaction NL-Z2.The pairing correlation is treated within the c...The ground state properties of Ds(Z=110) isotopes(N=151-195) are studied in the framework of the relativistic mean field(RMF) theory with the effective interaction NL-Z2.The pairing correlation is treated within the conventional BCS approximation.The calculated binding energies are consistent with the results from finite-range droplet model(FRDM) and Macroscopic-microscopic method(MMM).The quadrupole deformation,α-decay energy,α-decay half-live,charge radius,two-neutron separation energy and single-particle spectra are analyzed for Ds isotopes to find new characteristics of superheavy nuclei(SHN).Among the calculated results it is rather distinct that the isotopic shift appears evidently at neutron number N=184.展开更多
文摘[Objective] The purpose of this study was to construct a vscC in-frame deletion mutant of Vibrio alginolyticus with no antibiotic resistance marker. [Method] The first vscC mutant molecules in vitro were generated by SOE-PCR and then lig- ated to a suicide vector pDM4 to construct a suicide recombinant vector pDM4- A vscC. To clone the recombinant vector, it was transformed to E. coli SY327 strain, and then positive clones were selected and proved by PCR analysis. After that, the pDM4-AvscC DNA was extracted in large numbers and transformed to the E. coil S17-1 strain that acted as a donor in bacterial conjugation using the heat shock method. The recombinant E. coli S17-1 strains then transferred the pDM4-AvscC to V. alginolyticus ZJ51-O by conjugation method; transconjugants were screened and selected sequentially using antibiotic selection strategy and sucrose based counter-selection system to find the suspected mutants wanted. Finally the putative mutants were identified by PCR and confirmed by sequencing analysis. [Result] ZJ51-OAvscC was successfully constructed. [Conclusion] This study laid foundation for further research on the function of vscC gene and molecular mechanism of type Ⅲ secretion system in V. alginolyticus. Simultaneously, by the effective method other unknown functional genes in V. alginolyticus genome would be researched.
文摘The presence of DNA homologues corresponding to verc203 (vernalization-related cDNA clone) was investigated by molecular hybridization techniques. The genes were detected in 16 plant species that cover 12 subclasses of the Takhtajan system of angiosperms classification including diverse model species. The results of Southern blot analysis showed a low copy number of this gene existed in rice, wheat, barley and Arabidopsis. The hybridization result of PCR products demonstrated the conservation of the gene corresponding to ver203 in diverse plants. The phylogenetic tree of the ver203 gene in tested plants was supported by evolution relationship of species. The ver203 gene expressed in a vernalized plumule winter wheat, instead of the root. And the endosperm before the treatment was essential for the ver203 expression during vernalization in wheat. In Arabidopsis thaliana, the pattern of expression showed that the gene corresponding to ver203 was expressed at low temperature for 14 days. Gibberellin(GA3) may accelerate the expression of ver203 gene in Arabidopsis exposed to low temperature. However, it could not replace vernalization treatment to initiate the gene expression.
基金Foundation item: Projects(51108197, 51205215) supported by the National Natural Science Foundation of ChinaProjects(2011J05135, 2011J01318) supported by the Natural Science Foundation of Fujian Province, China+1 种基金Project(11QZR08) supported by the Scientific Research Foundation of Overseas Chinese Affairs Office of the State Council,ChinaProject(10BS213) supported by the Scientific Research Foundation for Advanced Talents,Huaqiao University,China
文摘Matrix effects can significantly hamper the accuracy and precision of the analysis results of perfluorinated acids (PFAs) in environmental solid samples. Several methods, such as standard addition, isotopically labeled internal standards, clean-up of SPE (solid phase extraction) eluents by dispersive graphitized carbon sorbent and substitution of eletrospray ionization (ESI) source by atmosphere pressure photoionization (APPI) source, were demonstrated for elimination of matrix effects in quantitative analysis of PFAs in solid samples. The resuRs indicate that matrix effects can be effectively eliminated by standard addition, but instrumental analysis time will be multiplied. Isotopically labeled internal standards can effectively negate matrix effects of PFAs with the same perfluorocarbon chain length, but is not valid for the other analytes. Although APPI can eliminate matrix effects for all analytes, it is only suitable for analysis of high pollution levels samples. Clean-up of SPE eluents by dispersive graphitized carbon sorbent not only effectively negate the impact of matrix effect, but also avoid frequent clean of the ESI in order to maintain instrumental sensitivity. Therefore, the best method for elimination of matrix effects is the usage of dispersive graphitized carbon sorbent for clean-up of SPE elution.
基金Supported by Shanghai Momentous Program,Grant No.07dz19505Shanghai Rising-star Program from the Science and Technology Commission of Shanghai Municipality,No.10QA1401400,China
文摘AIM:To develop lymph node metastasis(LNM)-associated biomarkers for colorectal cancer(CRC) using quantitative proteome analysis.METHODS:Differences in protein expression between primary CRC with LNM(LNM CRC) and without LNM(non-LNM CRC) were assessed using methyl esterification stable isotope labeling coupled with 2D liquid chromatography followed by tandem mass spectrometry(2D-LC-MS/MS).The relationship to clinicopathological parameters and prognosis of candidate biomarkers was examined using an independent sample set.RESULTS:Forty-three proteins were found to be differentially expressed by at least 2.5-fold in two types of CRC.S100A4 was significantly upregulated in LNM CRC compared with non-LNM CRC,which was confirmed by Western blotting,immunohistochemistry and real-time quantitative polymerase chain reaction.Further immunohistochemistry on another 112 CRC cases showed that overexpression of S100A4 frequently existed in LNM CRC compared with non-LNM CRC(P < 0.001).Overexpression of S100A4 was significantly associated with LNM(P < 0.001),advanced TNM stage(P < 0.001),increased 5-year recurrence rate(P < 0.001) and decreased 5-year overall survival rate(P < 0.001).Univariate and multivariate analyses indicated that S100A4 expression was an independent prognostic factor for recurrence and survival of CRC patients(P < 0.05).CONCLUSION:S100A4 might serve as a powerful biomarker for LNM and a prognostic factor in CRC.
文摘Objective : To evaluate the changes of serum matrix metalloproteinase-9 (romp-9) in patients of incipient diabetic nephropathy with or without macrovascular disease and to analyze the factors associated with homocysteine(hcy), interleukin-6(IL-6), tumor necrosis factor-alpha (TNF-α), highly sensitive C-reactive protein (hsCRP), HbAlc and lipid profile in those patients in order to know whether this marker or other factors are more important to induce diabetic macrovascular disease. Methods: Type 2 diabetes mellitus(T2DM) subjects with incipient diabetic nephropathy with or without macrovascular disease were selected for participation and divided into 2 groups. The patients in group 1 (n= 38) used insulin, and patients in group 2 (n=34) were treated with an oral antidiabetic drug. Then serum mmp-9, hey, IL-6 and TNF-α in these patients were measured, and compared to the healthy subjects as control (n= 16). The results were analyzed by SPSS13. Results: Serum romp-9 and hcy of the patients having incipient diabetic nephropathy with macrovascular disease were higher than that of patients without macrovascular disease (P〈0.01). For insulin-injected patients, whether they accompanied with macrovascular diseases or not, the serum levels of romp-9, hcy, IL-6 and TNF-α were all lower, but no significant statistics compared with non-insulin used patients or the healthy subjects. The serum level of romp-9 was more correlated with the serum hcy in antidiabetic drug used patients. (P〈0. 000) Conclusion: The serum level of romp-9 plays an important role of pathogenesis in the macrovascular disease in the incipient diabetic patients, and the serum level of hcy also can reflect the severely degree of macrovascular disease in these patients, insulin can reduce these markers.
基金supported by the National Natural Science Foundation of China(32170224,32225005)the NSFC-ERC International Cooperation and Exchange Programs(32311530331)the Youth Innovation Promotion Association CAS(2020391).
文摘Global climate change poses a severe threat to mountain biodiversity.Phenotypic plasticity and local adaptation are two common strategies for alpine plant to cope with such change.They may facilitate organismal adaptation to contrasting environments,depending on the influences of the environment or genotype or their interacted effects.In this study,we use an endemic alpine plant(Rorippa elata)in the Hengduan mountains(HDM)to unravel its phenotypic basis of adaptation strategy and evaluate the relative contributions of environment and genotype to its phenotype.We transplanted 37 genotypes of R.elata into two common gardens across low and high elevations(2800 vs.3800 m)during 2021-2022.Nine fitness-related traits were measured,including flowering probability and glucosinolates(GS)content.We estimated the environmental or genotypic contributions to the phenotype and identified the main environmental components.Our results revealed that both environment and genotype-by-environment interactions contributed to the phenotypes of R.elata.Latitudinal heterogeneity was identified as a key factor that explained 24%of the total phenotypic variation.In particular,genotypes of the northern HDM showed significantly higher plasticity in flowering probability than those of the southern HDM.Furthermore,within the southern HDM,GS content indicated local adaptation to herbivory stresses for R.elata genotypes along elevations.In conclusion,our results suggest that R.elata may have adapted to the alpine environment through species-level plasticity or regional-level local adaptation.These processes were shaped by either complex topography or interactions between genotype and mountain environments.Our study provides empirical evidence on the adaptation of alpine plants.
基金supported by the Strategic Priority Research Program of the Chinese Academy of Sciences(Grant No.XDA05090105)the R&D Special Fund for Public Welfare Industry(Meteorology)(Grant No.GYHY201206013)the National Key Technology R&D program(Grant No.2012BAC22B04)
文摘Long-term meteorological observation series are fundamental for reflecting climate changes.However,almost all meteorological stations inevitably undergo relocation or changes in observation instruments,rules,and methods,which can result in systematic biases in the observation series for corresponding periods.Homogenization is a technique for adjusting these biases in order to assess the true trends in the time series.In recent years,homogenization has shifted its focus from the adjustments to climate mean status to the adjustments to information about climate extremes or extreme weather.Using case analyses of ideal and actual climate series,here we demonstrate the basic idea of homogenization,introduce new understanding obtained from recent studies of homogenization of climate series in China,and raise issues for further studies in this field,especially with regards to climate extremes,uncertainty of the statistical adjustments,and biased physical relationships among different climate variables due to adjustments in single variable series.
基金Supported by the National Natural Science Foundation of China under Grant Nos. 10875152,10775061,10825522,and 10975064,11105035,11075066,and 11175074the Fundamental Research Funds for the Central Universities under Grant No. lzujbky-2012-5
文摘The ground state properties of Ds(Z=110) isotopes(N=151-195) are studied in the framework of the relativistic mean field(RMF) theory with the effective interaction NL-Z2.The pairing correlation is treated within the conventional BCS approximation.The calculated binding energies are consistent with the results from finite-range droplet model(FRDM) and Macroscopic-microscopic method(MMM).The quadrupole deformation,α-decay energy,α-decay half-live,charge radius,two-neutron separation energy and single-particle spectra are analyzed for Ds isotopes to find new characteristics of superheavy nuclei(SHN).Among the calculated results it is rather distinct that the isotopic shift appears evidently at neutron number N=184.