LeSPL-CNR is a crucial transcription factor for fruit ripening of Solanum lycopersicum. The cnr (colorless non-ripening) epimutation resulted from hypermethylation in a 286 bp region of LeSPL-CNR promoter inhibits nor...LeSPL-CNR is a crucial transcription factor for fruit ripening of Solanum lycopersicum. The cnr (colorless non-ripening) epimutation resulted from hypermethylation in a 286 bp region of LeSPL-CNR promoter inhibits normal fruit ripening. In present study, potential regulators of LeSPL-CNR, which could bind to the specific 286 bp region, were screened via south-western blotting and yeast one-hybrid (Y1H) library screening system. Results indicated that a total of 13 and 19 candidate proteins were acquired respectively, and both ribulose-1,5-bisphosphate carboxylase/oxygenase and 40S ribosomal protein were identified by two methods. These would provide some information for revealing roles of DNA methylation and the regulatory mechanism for LeSPL-CNR.展开更多
Blotting is a common technique widely used for molecular analysis in life sciences. The Western blot, in particular, is a process of transferring protein samples from a polyacrylamide gel to a blotting membrane and de...Blotting is a common technique widely used for molecular analysis in life sciences. The Western blot, in particular, is a process of transferring protein samples from a polyacrylamide gel to a blotting membrane and detecting the levels of specific proteins through reactions with primary and secondary antibodies. The state-of-the-art of Western blotting usually generates one blotting membrane per gel. However, multiple copies of blots are useful in many applications. Two blotting copies from a single protein gel, for instance, can be used for identifying a total amount of proteins of interest as well as its specific subpopulation level such as a phosphorylated isoform. To achieve this multi-blotting operation from a single gel, we modified a blotting procedure and developed a novel blotting device. The device consisted of a multi-anode plate and a microcontroller. It was designed to generate a well-controlled electrophoretic voltage profile, which allowed a quasi-uniform transfer of proteins of any size. The prototype device was built and its operation procedure was described. The experimental results clearly supported the notion that the described device was able to achieve multiple blotting from a single gel and reduce time and cost for protein analysis.展开更多
Recently, serum Golgi protein 73 （GP73） levels have been found to be elevated in patients with hepatocellu- lar carcinoma （HCC）, and GP73 has been proposed as a novel marker for HCC. However, GP73 levels in patien...Recently, serum Golgi protein 73 （GP73） levels have been found to be elevated in patients with hepatocellu- lar carcinoma （HCC）, and GP73 has been proposed as a novel marker for HCC. However, GP73 levels in patients remain controversial due to the specificity of the anti-GP73 antibody-based enzyme linked immunosorbent as- say （ELISA）. Therefore, an anti-GP73 antibody with high specificity was highly demanded. In the present study, by hybridoma screening, we generated an anti-GP73 monoclonal antibody （mAb） designated as 6A2 using recom- binant GP73 protein produced by prokaryotic expression. The specificity of 6A2 was evaluated by Western blot- ting, immunohistochemistry and immunoprecipitation. The results showed that 6A2 recognized GP73 in both native and denatured forms. In addition, we have developed a sandwich ELISA using 6A2 and GP73 polyclonal antibody generated in New Zealand white rabbits according to standard procedures, and measured the serum GP73 level of patients using this assay. Our results showed that serum GP73 levels of HCC patients were significantly higher than those of healthy controls （P = 0.0036）. Furthermore, for the first time, GP73 serum level was found to be elevated in patients with breast cancer compared with healthy controls （P = 0.0172）.展开更多
Based on the Regional Specialized Meteorological Center(RSMC)Tokyo-Typhoon Center best-track data and the NCEP-NCAR reanalysis dataset,extratropical transitioning(ET)tropical cyclones(ETCs)over the western North Pacif...Based on the Regional Specialized Meteorological Center(RSMC)Tokyo-Typhoon Center best-track data and the NCEP-NCAR reanalysis dataset,extratropical transitioning(ET)tropical cyclones(ETCs)over the western North Pacific(WNP)during 1951–2021 are classified into six clusters using the fuzzy c-means clustering method(FCM)according to their track patterns.The characteristics of the six hard-clustered ETCs with the highest membership coefficient are shown.Most tropical cyclones(TCs)that were assigned to clusters C2,C5,and C6 made landfall over eastern Asian countries,which severely threatened these regions.Among landfalling TCs,93.2%completed their ET after landfall,whereas 39.8%of ETCs completed their transition within one day.The frequency of ETCs over the WNP has decreased in the past four decades,wherein cluster C5 demonstrated a significant decrease on both interannual and interdecadal timescales with the expansion and intensification of the western Pacific subtropical high(WPSH).This large-scale circulation pattern is favorable for C2 and causes it to become the dominant track pattern,owning to it containing the largest number of intensifying ETCs among the six clusters,a number that has increased insignificantly over the past four decades.The surface roughness variation and three-dimensional background circulation led to C5 containing the maximum number of landfalling TCs and a minimum number of intensifying ETCs.Our results will facilitate a better understanding of the spatiotemporal distributions of ET events and associated environment background fields,which will benefit the effective monitoring of these events over the WNP.展开更多
Based on an analysis of the limitations of conventional production component methods for natural gas development planning,this study proposes a new one that uses life cycle models for the trend fitting and prediction ...Based on an analysis of the limitations of conventional production component methods for natural gas development planning,this study proposes a new one that uses life cycle models for the trend fitting and prediction of production.In this new method,the annual production of old and new wells is predicted by year first and then is summed up to yield the production for the planning period.It shows that the changes in the production of old wells in old blocks can be fitted and predicted using the vapor pressure model(VPM),with precision of 80%e95%,which is 6.6%e13.2%higher than that of other life cycle models.Furthermore,a new production prediction process and method for new wells have been established based on this life cycle model to predict the production of medium-to-shallow gas reservoirs in western Sichuan Basin,with predication error of production rate in 2021 and 2022 being 6%and 3%respectively.The new method can be used to guide the medium-and long-term planning or annual scheme preparation for gas development.It is also applicable to planning for large single gas blocks that require continuous infill drilling and adjustment to improve gas recovery.展开更多
BACKGROUND Serologic cross-reactivity between hantaviruses often complicates the interpretation of the results.AIM To analyze the diagnostic value of indirect immunofluorescence assay(IFA)and western blot(WB)in the di...BACKGROUND Serologic cross-reactivity between hantaviruses often complicates the interpretation of the results.AIM To analyze the diagnostic value of indirect immunofluorescence assay(IFA)and western blot(WB)in the diagnosis of hantavirus infections.METHODS One hundred eighty-eight serum samples from Puumala(PUUV)and Dobrava(DOBV)orthohantavirus infected patients were analyzed.Serology was performed using commercial tests(Euroimmun,Lübeck,Germany).RESULTS Using IFA,49.5%of acute-phase samples showed a monotypic response to PUUV,while 50.5% cross-reacted with other hantaviruses.The overall cross-reactivity was higher for immunoglobulin G(IgG)(50.0%)than for immunoglobulin M(IgM)(25.5%).PUUV IgM/IgG antibodies showed low/moderate reactivity with orthohantaviruses Hantaan(12.3%/31.5%),Seoul(7.5%/17.8%),DOBV(5.4%/28.1%),and Saaremaa(4.8%/15.7%).Both DOBV IgM and IgG antibodies were broadly reactive with Hantaan(76.2%/95.2%),Saaremaa(80.9%/83.3%),and Seoul(78.6%/85.7%)and moderate with PUUV(28.5%/38.1%).Using a WB,serotyping was successful in most cross-reactive samples(89.5%).CONCLUSION The presented results indicate that WB is more specific than IFA in the diagnosis of hantavirus infections,confirming serotype in most IFA cross-reactive samples.展开更多
文摘LeSPL-CNR is a crucial transcription factor for fruit ripening of Solanum lycopersicum. The cnr (colorless non-ripening) epimutation resulted from hypermethylation in a 286 bp region of LeSPL-CNR promoter inhibits normal fruit ripening. In present study, potential regulators of LeSPL-CNR, which could bind to the specific 286 bp region, were screened via south-western blotting and yeast one-hybrid (Y1H) library screening system. Results indicated that a total of 13 and 19 candidate proteins were acquired respectively, and both ribulose-1,5-bisphosphate carboxylase/oxygenase and 40S ribosomal protein were identified by two methods. These would provide some information for revealing roles of DNA methylation and the regulatory mechanism for LeSPL-CNR.
文摘Blotting is a common technique widely used for molecular analysis in life sciences. The Western blot, in particular, is a process of transferring protein samples from a polyacrylamide gel to a blotting membrane and detecting the levels of specific proteins through reactions with primary and secondary antibodies. The state-of-the-art of Western blotting usually generates one blotting membrane per gel. However, multiple copies of blots are useful in many applications. Two blotting copies from a single protein gel, for instance, can be used for identifying a total amount of proteins of interest as well as its specific subpopulation level such as a phosphorylated isoform. To achieve this multi-blotting operation from a single gel, we modified a blotting procedure and developed a novel blotting device. The device consisted of a multi-anode plate and a microcontroller. It was designed to generate a well-controlled electrophoretic voltage profile, which allowed a quasi-uniform transfer of proteins of any size. The prototype device was built and its operation procedure was described. The experimental results clearly supported the notion that the described device was able to achieve multiple blotting from a single gel and reduce time and cost for protein analysis.
基金supported by the Science and Technology Foundation of Nanjing Medical University (No. 07NMUZ005).
文摘Recently, serum Golgi protein 73 （GP73） levels have been found to be elevated in patients with hepatocellu- lar carcinoma （HCC）, and GP73 has been proposed as a novel marker for HCC. However, GP73 levels in patients remain controversial due to the specificity of the anti-GP73 antibody-based enzyme linked immunosorbent as- say （ELISA）. Therefore, an anti-GP73 antibody with high specificity was highly demanded. In the present study, by hybridoma screening, we generated an anti-GP73 monoclonal antibody （mAb） designated as 6A2 using recom- binant GP73 protein produced by prokaryotic expression. The specificity of 6A2 was evaluated by Western blot- ting, immunohistochemistry and immunoprecipitation. The results showed that 6A2 recognized GP73 in both native and denatured forms. In addition, we have developed a sandwich ELISA using 6A2 and GP73 polyclonal antibody generated in New Zealand white rabbits according to standard procedures, and measured the serum GP73 level of patients using this assay. Our results showed that serum GP73 levels of HCC patients were significantly higher than those of healthy controls （P = 0.0036）. Furthermore, for the first time, GP73 serum level was found to be elevated in patients with breast cancer compared with healthy controls （P = 0.0172）.
基金supported by the National Natural Science Foundation of China(Grant Nos.42075053 and 41975128)。
文摘Based on the Regional Specialized Meteorological Center(RSMC)Tokyo-Typhoon Center best-track data and the NCEP-NCAR reanalysis dataset,extratropical transitioning(ET)tropical cyclones(ETCs)over the western North Pacific(WNP)during 1951–2021 are classified into six clusters using the fuzzy c-means clustering method(FCM)according to their track patterns.The characteristics of the six hard-clustered ETCs with the highest membership coefficient are shown.Most tropical cyclones(TCs)that were assigned to clusters C2,C5,and C6 made landfall over eastern Asian countries,which severely threatened these regions.Among landfalling TCs,93.2%completed their ET after landfall,whereas 39.8%of ETCs completed their transition within one day.The frequency of ETCs over the WNP has decreased in the past four decades,wherein cluster C5 demonstrated a significant decrease on both interannual and interdecadal timescales with the expansion and intensification of the western Pacific subtropical high(WPSH).This large-scale circulation pattern is favorable for C2 and causes it to become the dominant track pattern,owning to it containing the largest number of intensifying ETCs among the six clusters,a number that has increased insignificantly over the past four decades.The surface roughness variation and three-dimensional background circulation led to C5 containing the maximum number of landfalling TCs and a minimum number of intensifying ETCs.Our results will facilitate a better understanding of the spatiotemporal distributions of ET events and associated environment background fields,which will benefit the effective monitoring of these events over the WNP.
基金funded by the project entitled Technical Countermeasures for the Quantitative Characterization and Adjustment of Residual Gas in Tight Sandstone Gas Reservoirs of the Daniudi Gas Field(P20065-1)organized by the Science&Technology R&D Department of Sinopec.
文摘Based on an analysis of the limitations of conventional production component methods for natural gas development planning,this study proposes a new one that uses life cycle models for the trend fitting and prediction of production.In this new method,the annual production of old and new wells is predicted by year first and then is summed up to yield the production for the planning period.It shows that the changes in the production of old wells in old blocks can be fitted and predicted using the vapor pressure model(VPM),with precision of 80%e95%,which is 6.6%e13.2%higher than that of other life cycle models.Furthermore,a new production prediction process and method for new wells have been established based on this life cycle model to predict the production of medium-to-shallow gas reservoirs in western Sichuan Basin,with predication error of production rate in 2021 and 2022 being 6%and 3%respectively.The new method can be used to guide the medium-and long-term planning or annual scheme preparation for gas development.It is also applicable to planning for large single gas blocks that require continuous infill drilling and adjustment to improve gas recovery.
文摘BACKGROUND Serologic cross-reactivity between hantaviruses often complicates the interpretation of the results.AIM To analyze the diagnostic value of indirect immunofluorescence assay(IFA)and western blot(WB)in the diagnosis of hantavirus infections.METHODS One hundred eighty-eight serum samples from Puumala(PUUV)and Dobrava(DOBV)orthohantavirus infected patients were analyzed.Serology was performed using commercial tests(Euroimmun,Lübeck,Germany).RESULTS Using IFA,49.5%of acute-phase samples showed a monotypic response to PUUV,while 50.5% cross-reacted with other hantaviruses.The overall cross-reactivity was higher for immunoglobulin G(IgG)(50.0%)than for immunoglobulin M(IgM)(25.5%).PUUV IgM/IgG antibodies showed low/moderate reactivity with orthohantaviruses Hantaan(12.3%/31.5%),Seoul(7.5%/17.8%),DOBV(5.4%/28.1%),and Saaremaa(4.8%/15.7%).Both DOBV IgM and IgG antibodies were broadly reactive with Hantaan(76.2%/95.2%),Saaremaa(80.9%/83.3%),and Seoul(78.6%/85.7%)and moderate with PUUV(28.5%/38.1%).Using a WB,serotyping was successful in most cross-reactive samples(89.5%).CONCLUSION The presented results indicate that WB is more specific than IFA in the diagnosis of hantavirus infections,confirming serotype in most IFA cross-reactive samples.
