Objective： S100A6 （a.k.a., calcyclin） is over-expressed in several human tumors, including gastric carcinoma, human melanoma, pancreatic carcinoma, squamous cell carcinoma, malignant fibrous histiocytoma （MFH）, a...Objective： S100A6 （a.k.a., calcyclin） is over-expressed in several human tumors, including gastric carcinoma, human melanoma, pancreatic carcinoma, squamous cell carcinoma, malignant fibrous histiocytoma （MFH）, and carcinomas of the thyroid, breast, and colon. However, little is known about the role S100A6 plays in gastric adenocarcinoma. In the present study, we intended to investigate the influence of S100A6 on the growth, proliferation, apoptosis, invasion and cell cycle of the gastric cancer cell MKN45. Methods： As an important member of S100 family, S100A6 cDNAwas subcloned into a constitutive vector pcDNA3.1 followed by transfection in gastric cancer cell line MKN45 by using liposome. Then stable transfectants were selected and appraised. The apoptosis and cell cycles of these clones were analyzed by using flow cytometric assay. The growth and proliferation were analyzed by cell growth curves and colony-forming assay respectively. The S100A6 stable expression clones （MKN-S100A6） were detected and compared with their control groups respectively. Results： MKN- S100A6 grew faster than MKN45 and MKN-PC （MKN45 transfected with pcDNA3.1 vector）. The cell counts of MKN-SI00A6 in the fifth, sixth and seventh days were significantly more than those of control groups （P 〈 0.05）. Cell cycle analysis showed that proportions of MKN-S100A6 in G0-G1 and G2-M were different significantly with those of its control groups respectively （P 〈 0.05）. The apoptosis rate of MKN-S100A6 was significantly lower than those of control groups （P 〈 0.05）. Results of colony-forming assay showed that the colon formation rate of MKN-S100A6 was higher than those of control groups （P 〈 0.05）. Conclusion： S100A6 can promote the growth and proliferation of gastric cancer cells. It can help tumor cell maintain malignant phenotype. In gastric cancer, S100A6 could be thought as a tumor-enhancing gene in some distance, but its role could be complicated.展开更多
Background:Whether adherence to a healthy lifestyle is associated with a lower risk of developing pneumonia and a better long-term prognosis remains unclear.This study aimed to investigate associations of individual a...Background:Whether adherence to a healthy lifestyle is associated with a lower risk of developing pneumonia and a better long-term prognosis remains unclear.This study aimed to investigate associations of individual and combined lifestyle factors(LFs)with the incidence risk and long-term prognosis of pneumonia hospitalization.Methods:Using data from the China Kadoorie Biobank study,we used the multistate models to investigate the role of five high-risk LFs,including smoking,excessive alcohol drinking,unhealthy dietary habits,physical inactivity,and unhealthy body shape,alone or in combination in the transitions from a generally healthy state at baseline to pneumonia hospitalization or cardiovascular disease(CVD,regarded as a reference outcome),and subsequently to mortality.Results:Most of the five high-risk LFs were associated with increased risks of transitions from baseline to pneumonia and from pneumonia to death,but with different risk estimates.The greater the number of high-risk LFs,the higher the risk of developing pneumonia and long-term mortality risk after pneumonia,with the strength of associations comparable to that of LFs and CVD.Compared to participants with 0-1 high-risk LF,the adjusted hazard ratios(HRs)and 95%confidence intervals(CIs)for transitions from baseline to pneumonia and from pneumonia to death in those with five high-risk LFs were 1.43(1.28-1.60)and 1.98(1.61-2.42),respectively.Correspondingly,the respective HRs(95%CIs)for transitions from baseline to CVD and from CVD to death were 2.00(1.89-2.11)and 1.44(1.30-1.59),respectively.The risk estimates changed slightly when further adjusting for the presence of major chronic diseases.Conclusion:In this Chinese population,unhealthy LFs were associated with an increased incidence and long-term mortality risk of pneumonia.展开更多
Coronaviruses can infect humans,mammals,and birds,leading to respiratory,gastrointestinal,and neurological diseases.These viruses are significant zoonotic pathogens with nine known types capable of infecting humans.Th...Coronaviruses can infect humans,mammals,and birds,leading to respiratory,gastrointestinal,and neurological diseases.These viruses are significant zoonotic pathogens with nine known types capable of infecting humans.The coronavirus genome,approximately 30 kb in size,is the largest known ribonucleic acid(RNA)virus genome,and its complexity makes assembly and manipulation time-consuming and labor-intensive.Reverse genetic systems are widely used to engineer recombinant viruses that can be adapted at Biosafety Level 2(BSL-2)for studying viral gene function,replication,pathogenesis,vaccines,and therapeutics.The infectious clones,which enabled the recovery of various viruses after DNA recombinant technology,were indispensable tools for the reverse genetics of viruses.Various techniques for constructing infectious clones of human coronaviruses(HCoV)have been developed,encompassing methods such as vaccinia virus vectors method,in vitro ligation,bacterial artificial chromosome systems,yeast artificial chromosome systems,circular polymerase extension reaction,and the recently reported infectious sub-genomic amplicons technology.This review summarizes the status of various techniques for constructing infectious clones of human coronaviruses and related applications.展开更多
Viral infectious clones(ICs)serve as robust platforms for studying viral biology and screening antiviral agents using reverse genetics.However,the molecular profiles and complex limitations of human coronaviruses(HCoV...Viral infectious clones(ICs)serve as robust platforms for studying viral biology and screening antiviral agents using reverse genetics.However,the molecular profiles and complex limitations of human coronaviruses(HCoVs)pose a challenge to ICs development.In this study,we report a novel platform to develop the ICs for HCoV-OC43-VR1558 using a one-step assembly method in yeast by transformation-associated recombination(TAR)technology.Recombinant HCoV-OC43-VR1558,named as rOC43(1558)-WT,was rapidly generated by TAR.In addition,recombinant HCoV-OC43-VR1558-expressing reporter genes,named as rOC43(1558)-ns2FusionRluc,was also generated based on TAR by inserting the ns2 region of the IC with Renilla luciferase(Rluc).We further characterized their replication through virus titration using 50%tissue culture infective dose(TCID50)and indirect immunofluorescence assay(IFA),luciferase reporter assay,and western blotting(WB)assay.The genetic stability of the recombinant HCoV-OC43 was assessed through viral genome sequencing following passaging in BHK-21 cells.These reporter viruses were validated as screening tools for inhibitorsin vitro by evaluating the antiviral activities of remdesivir and chloroquine.The phenotypes of HCoV-OC43-VR1558 and HCoV-OC43-VR759 were comparedin vitro andin vivo.The TAR-based one-step assembly of IC was successfully applied,facilitating the rapid generation of recombinant HCoV-OC43 and providing a useful platform for the investigation of biological mechanisms,development of vaccines and diagnostic tests,and screening inhibitors of HCoVs.展开更多
基金Supported by a grant from the National Natural Science Foundation of China (No: 30600728)
文摘Objective： S100A6 （a.k.a., calcyclin） is over-expressed in several human tumors, including gastric carcinoma, human melanoma, pancreatic carcinoma, squamous cell carcinoma, malignant fibrous histiocytoma （MFH）, and carcinomas of the thyroid, breast, and colon. However, little is known about the role S100A6 plays in gastric adenocarcinoma. In the present study, we intended to investigate the influence of S100A6 on the growth, proliferation, apoptosis, invasion and cell cycle of the gastric cancer cell MKN45. Methods： As an important member of S100 family, S100A6 cDNAwas subcloned into a constitutive vector pcDNA3.1 followed by transfection in gastric cancer cell line MKN45 by using liposome. Then stable transfectants were selected and appraised. The apoptosis and cell cycles of these clones were analyzed by using flow cytometric assay. The growth and proliferation were analyzed by cell growth curves and colony-forming assay respectively. The S100A6 stable expression clones （MKN-S100A6） were detected and compared with their control groups respectively. Results： MKN- S100A6 grew faster than MKN45 and MKN-PC （MKN45 transfected with pcDNA3.1 vector）. The cell counts of MKN-SI00A6 in the fifth, sixth and seventh days were significantly more than those of control groups （P 〈 0.05）. Cell cycle analysis showed that proportions of MKN-S100A6 in G0-G1 and G2-M were different significantly with those of its control groups respectively （P 〈 0.05）. The apoptosis rate of MKN-S100A6 was significantly lower than those of control groups （P 〈 0.05）. Results of colony-forming assay showed that the colon formation rate of MKN-S100A6 was higher than those of control groups （P 〈 0.05）. Conclusion： S100A6 can promote the growth and proliferation of gastric cancer cells. It can help tumor cell maintain malignant phenotype. In gastric cancer, S100A6 could be thought as a tumor-enhancing gene in some distance, but its role could be complicated.
基金This work was supported by grants from the National Natural Science Foundation of China(Nos.82388102,82192904,82404336,and 82192900)the Noncommunicable Chronic Diseases-National Science and Technology Major Project(No.2023ZD0510100)+4 种基金the Kadoorie Charitable Foundation in Hong Kong,the UK Wellcome Trust(Nos.212946/Z/18/Z,202922/Z/16/Z,104085/Z/14/Z,and 088158/Z/09/Z)the National Key R&D Program of China(No.2016YFC0900500)the National Natural Science Foundation of China(Nos.81390540,91846303,and 81941018)the Chinese Ministry of Science and Technology(No.2011BAI09B01)the High-Level Talents Research Start-Up Project of Fujian Medical University(No.XRCZX2023014).
文摘Background:Whether adherence to a healthy lifestyle is associated with a lower risk of developing pneumonia and a better long-term prognosis remains unclear.This study aimed to investigate associations of individual and combined lifestyle factors(LFs)with the incidence risk and long-term prognosis of pneumonia hospitalization.Methods:Using data from the China Kadoorie Biobank study,we used the multistate models to investigate the role of five high-risk LFs,including smoking,excessive alcohol drinking,unhealthy dietary habits,physical inactivity,and unhealthy body shape,alone or in combination in the transitions from a generally healthy state at baseline to pneumonia hospitalization or cardiovascular disease(CVD,regarded as a reference outcome),and subsequently to mortality.Results:Most of the five high-risk LFs were associated with increased risks of transitions from baseline to pneumonia and from pneumonia to death,but with different risk estimates.The greater the number of high-risk LFs,the higher the risk of developing pneumonia and long-term mortality risk after pneumonia,with the strength of associations comparable to that of LFs and CVD.Compared to participants with 0-1 high-risk LF,the adjusted hazard ratios(HRs)and 95%confidence intervals(CIs)for transitions from baseline to pneumonia and from pneumonia to death in those with five high-risk LFs were 1.43(1.28-1.60)and 1.98(1.61-2.42),respectively.Correspondingly,the respective HRs(95%CIs)for transitions from baseline to CVD and from CVD to death were 2.00(1.89-2.11)and 1.44(1.30-1.59),respectively.The risk estimates changed slightly when further adjusting for the presence of major chronic diseases.Conclusion:In this Chinese population,unhealthy LFs were associated with an increased incidence and long-term mortality risk of pneumonia.
基金the National Key Research and Development Program of China(2022YFC2304101,2021YFA1201003).
文摘Coronaviruses can infect humans,mammals,and birds,leading to respiratory,gastrointestinal,and neurological diseases.These viruses are significant zoonotic pathogens with nine known types capable of infecting humans.The coronavirus genome,approximately 30 kb in size,is the largest known ribonucleic acid(RNA)virus genome,and its complexity makes assembly and manipulation time-consuming and labor-intensive.Reverse genetic systems are widely used to engineer recombinant viruses that can be adapted at Biosafety Level 2(BSL-2)for studying viral gene function,replication,pathogenesis,vaccines,and therapeutics.The infectious clones,which enabled the recovery of various viruses after DNA recombinant technology,were indispensable tools for the reverse genetics of viruses.Various techniques for constructing infectious clones of human coronaviruses(HCoV)have been developed,encompassing methods such as vaccinia virus vectors method,in vitro ligation,bacterial artificial chromosome systems,yeast artificial chromosome systems,circular polymerase extension reaction,and the recently reported infectious sub-genomic amplicons technology.This review summarizes the status of various techniques for constructing infectious clones of human coronaviruses and related applications.
基金supported by the National Key Research and Development Program of China(2022YFC2304100 and 2021YFA1201003).
文摘Viral infectious clones(ICs)serve as robust platforms for studying viral biology and screening antiviral agents using reverse genetics.However,the molecular profiles and complex limitations of human coronaviruses(HCoVs)pose a challenge to ICs development.In this study,we report a novel platform to develop the ICs for HCoV-OC43-VR1558 using a one-step assembly method in yeast by transformation-associated recombination(TAR)technology.Recombinant HCoV-OC43-VR1558,named as rOC43(1558)-WT,was rapidly generated by TAR.In addition,recombinant HCoV-OC43-VR1558-expressing reporter genes,named as rOC43(1558)-ns2FusionRluc,was also generated based on TAR by inserting the ns2 region of the IC with Renilla luciferase(Rluc).We further characterized their replication through virus titration using 50%tissue culture infective dose(TCID50)and indirect immunofluorescence assay(IFA),luciferase reporter assay,and western blotting(WB)assay.The genetic stability of the recombinant HCoV-OC43 was assessed through viral genome sequencing following passaging in BHK-21 cells.These reporter viruses were validated as screening tools for inhibitorsin vitro by evaluating the antiviral activities of remdesivir and chloroquine.The phenotypes of HCoV-OC43-VR1558 and HCoV-OC43-VR759 were comparedin vitro andin vivo.The TAR-based one-step assembly of IC was successfully applied,facilitating the rapid generation of recombinant HCoV-OC43 and providing a useful platform for the investigation of biological mechanisms,development of vaccines and diagnostic tests,and screening inhibitors of HCoVs.
