With the approval of more and more genetically modified(GM)crops in our country,GM safety management has become more important.Transgenic detection is a major approach for transgenic safety management.Nevertheless,a c...With the approval of more and more genetically modified(GM)crops in our country,GM safety management has become more important.Transgenic detection is a major approach for transgenic safety management.Nevertheless,a convenient and visual technique with low equipment requirements and high sensitivity for the field detection of GM plants is still lacking.On the basis of the existing recombinase polymerase amplification(RPA)technique,we developed a multiplex RPA(multi-RPA)method that can simultaneously detect three transgenic elements,including the cauliflower mosaic virus 35S gene(CaMV35S)promoter,neomycin phosphotransferaseⅡgene(NptⅡ)and hygromycin B phosphotransferase gene(Hyg),thus improving the detection rate.Moreover,we coupled this multi-RPA technique with the CRISPR/Cas12a reporter system,which enabled the detection results to be clearly observed by naked eyes under ultraviolet(UV)light(254 nm;which could be achieved by a portable UV flashlight),therefore establishing a multi-RPA visual detection technique.Compared with the traditional test strip detection method,this multi-RPA-CRISPR/Cas12a technique has the higher specificity,higher sensitivity,wider application range and lower cost.Compared with other polymerase chain reaction(PCR)techniques,it also has the advantages of low equipment requirements and visualization,making it a potentially feasible method for the field detection of GM plants.展开更多
The three-dimensional internal flow field of centrifugal pump is complex and variable with design parameters and operation conditions. The post-processing technique named differential amplification method was proposed...The three-dimensional internal flow field of centrifugal pump is complex and variable with design parameters and operation conditions. The post-processing technique named differential amplification method was proposed for the comparison study of different flow structures. The full steady flow fields of an industrial centrifugal pump working on-design and off-design points were numerically investigated by solving Reynolds average Navier-Stokes equations together with a shear-stress transport(SST) k-? turbulence model. And the numerically predicted performance curves of the studied pump agree well with test measurement results. Compared with the flow flied on design point under the help of differential amplification method, the disturbance caused by interaction between blade and volute tongue is very obvious and it extends to the diffuser pipe on the working point with 0.8 times rated flux. While on the point with 1.2 times rated flux, the flow distribution in impeller region is roughly even and it flows more to the bottom section of the diffuser pipe. The above method was proved to be good at displaying the subtle secondary flow structure changes with a higher resolution effect relative to single isolated case observation, which helps the optimization decision-making from multiple design cases.展开更多
The phenomenon of ground vibration amplification caused by railway traffic was found and proved. In order to study the reasons which cause the amplification, a drop-weight test was performed. Then, the model for both ...The phenomenon of ground vibration amplification caused by railway traffic was found and proved. In order to study the reasons which cause the amplification, a drop-weight test was performed. Then, the model for both homogeneous and layered soil subjected to a harmonic vertical load was built. With the help of this model, displacement Green's function was calculated and the propagation laws of ground vibration responses were discussed. Results show that: 1) When applying a harmonic load on the half-space surface, the amplitude of ground vibrations attenuate with fluctuation, which is caused by the superposition of bulk and Rayleigh waves. 2) Vibration amplification can be enlarged under the conditions of embedded source and the soil layers. 3) In practice, the fluctuant attenuation should be paid attention to especially for the vibration receivers who are sensitive to single low frequencies(<10 Hz). Moreover, for the case of embedded loads, it should also be paid attention to that the receivers are located at the place where the horizontal distance is similar to embedded depth, usually 10 to 30 m for metro lines.展开更多
In order to explore whether the rearrangement and amplification of C-erbB-l gene were associatedwith the development of MDS and their clinicalsignificance in diagnoses and prognoses. digitoxin-labelled oligonucleotide...In order to explore whether the rearrangement and amplification of C-erbB-l gene were associatedwith the development of MDS and their clinicalsignificance in diagnoses and prognoses. digitoxin-labelled oligonucleotide probes were used to detect展开更多
When the loop-mediated isothermal amplification(LAMP)assay is used for detecting target genes,DNA extraction is unnecessary in many cases.Simple pretreatment(e.g.heating)is enough to obtain rather sensitive responses....When the loop-mediated isothermal amplification(LAMP)assay is used for detecting target genes,DNA extraction is unnecessary in many cases.Simple pretreatment(e.g.heating)is enough to obtain rather sensitive responses.Even test samples without any pretreatment can be used as template.This feature suggests that LAMP is superior to PCR in developing point-of-care test strategies.In this study,using Stx1 gene from E.coli as model,we verified that viable cells,dead cells and extracellular DNA could function as template in the LAMP assay.In the incubation at 63℃,viable bacteria in the LAMP reaction mixture lysed completely within 2 min,providing DNA template for nucleic acid amplification.The Stx1 gene in diluted culture medium,spiked tap water,spiked seawater and real seawater all could be detected,with or without the step of DNA extraction.We found that the complex substances in real sample(e.g.natural seawater)exhibited considerable inhibitory effect on the sensitivity of the LAMP assay.These outcomes are meaningful for building a point-of-care strategy by employing the LAMP assay for environmental monitoring,bio-resource surveys,food safety,etc.in particular those based on environmental DNA.展开更多
Rice bacterial leaf brown spot disease caused by Pseudomonas syringae pv.syringae(Pss)is a major disease on rice.In recent years,Pss has emerged worldwide,seriously affecting rice production.It is very important to es...Rice bacterial leaf brown spot disease caused by Pseudomonas syringae pv.syringae(Pss)is a major disease on rice.In recent years,Pss has emerged worldwide,seriously affecting rice production.It is very important to establish a rapid detection method of Pss for the diagnosis and prevention of this disease.In order to robust and accurately diagnose the rice bacterial leaf brown spot disease in the field and laboratory,an assay system for the Pss was developed in this study,and the specific sequence of hrcN was used as the target,based on loop-mediated isothermal amplification(LAMP).The best detection system was MgSO 48 mmol·L^(-1),Bst DNA polymerase 8 U,dNTP 1.4 mmol·L^(-1),the ratio of internal and outer primers was 2:1,the reaction temperature was 63℃,the reaction time was 45 min,and the lowest sensitivity was 104 CFU·mL^(-1).This results provided an accurate and robust method for laboratory and field diagnosis of bacterial leaf brown spot disease of rice.展开更多
The availability of the plastid genome sequences is one of the bases for comparative,functional,and structural genomic studies of plastid-containing living organisms,in addition to the application
Soil DNA extraction,such as microbial community analysis and gene drift detection,is an important basis for multiple analyses in different fields.Nevertheless,the soil DNA extraction methods for field detection are st...Soil DNA extraction,such as microbial community analysis and gene drift detection,is an important basis for multiple analyses in different fields.Nevertheless,the soil DNA extraction methods for field detection are still lacking.This study established a rapid soil DNA extraction(RSDE)method that can be used in field detection.In this method,we first utilized the optimized lysate to isolate DNA from soil and then used a filtration membrane and a DNA adsorption membrane to purify the DNA via the column method.Moreover,we used the pressure from the syringe instead of the conventional centrifugal force of the centrifuge to assist the sample filtration,resulting in very low requirements for this method,with an extraction time of less than 20 min.Furthermore,we demonstrated that the RSDE method was applicable for DNA extraction from different types of soils,with the demand for soil samples as low as 0.1 g and that the amount of obtained DNA was,to some extent,greater than that obtained by a commercial kit.Further analysis revealed that this extracted genomic DNA can be used directly for polymerase chain reaction(PCR)analysis,including ordinary PCR,real-time fluorescent quantitative PCR,and recombinase polymerase amplification(RPA)-CRISPR/Cas12a visual assays.In addition,we demonstrated that this method can be used to extract DNA from residual plant roots in addition to soil microbes,which lays a foundation for the comprehensive analysis of soil plants and microorganisms.In summary,the RSDE method proposed in this study may have wide application prospects.展开更多
The seismic damage to ancillary facilities on high-speed railway(HSR)bridges can affect the normal movement of trains.To propose the bridge deck acceleration response spectra of the typical HSR simply-supported girder...The seismic damage to ancillary facilities on high-speed railway(HSR)bridges can affect the normal movement of trains.To propose the bridge deck acceleration response spectra of the typical HSR simply-supported girder bridge for simplifying the seismic responses analysis of the facilities on bridges,the finite element models of the HSR multi-span simply-supported girder bridges with CRTSII track were established,and the numerical model was validated by tests.Besides,the effects of the span number,peak ground acceleration(PGA),pier height on the seismic acceleration and response spectra of the bridge deck were investigated.Afterward,the bridge acceleration amplification factor curves and bridge deck response spectra with different PGAs and pier heights were obtained.The formula for bridge deck acceleration amplification factor,with a 95%guarantee rate,was fitted.Moreover,the finite element models of the overhead contact lines(OCL)mounted on rigid base and bridges were established to validate the fitted formula.The results indicated that the maximum seismic acceleration response is in the midspan of the beam.The proposed formula for the bridge deck acceleration response spectra can be used to analyze the earthquake response of the OCL and other ancillary facilities on HSR simply-supported girder bridges.The bridge deck acceleration response spectra are conservative in terms of structural safety and can significantly improving the analysis efficiency.展开更多
Objective The detection of RNA single nucleotide polymorphism(SNP)is of great importance due to their association with protein expression related to various diseases and drug responses.At present,splintR ligase-assist...Objective The detection of RNA single nucleotide polymorphism(SNP)is of great importance due to their association with protein expression related to various diseases and drug responses.At present,splintR ligase-assisted methods are important approaches for RNA direct detection,but its specificity will be limited when the fidelity of ligases is not ideal.The aim of this study was to create a method to improve the specificity of splintR ligase for RNA detection.Methods In this study,a dualcompetitive-padlock-probe(DCPLP)assay without the need for additional enzymes or reactions is proposed to improve specificity of splintR ligase ligation.To verify the method,we employed dual competitive padlock probe-mediated rolling circle amplification(DCPLP-RCA)to genotype the CYP2C9 gene.Results The specificity was well improved through the competition and strand displacement of dual padlock probe,with an 83.26%reduction in nonspecific signal.By detecting synthetic RNA samples,the method demonstrated a dynamic detection range of 10 pmol/L-1 nmol/L.Furthermore,clinical samples were applied to the method to evaluate its performance,and the genotyping results were consistent with those obtained using the qPCR method.Conclusion This study has successfully established a highly specific direct RNA SNP detection method,and provided a novel avenue for accurate identification of various types of RNAs.展开更多
Past earthquakes have revealed that topographic features have significant impacts on the characteristics of ground motions,which may cause the amplification and de-amplification of input seismic waves.The topographic ...Past earthquakes have revealed that topographic features have significant impacts on the characteristics of ground motions,which may cause the amplification and de-amplification of input seismic waves.The topographic effect with the assumption of plane seismic waves on the seismic responses of bridges has been investigated in the existing literature;however,the influence of near-source topographic effects has not been thoroughly understood.The objective of this study is to numerically explore the near-source topographic effects on the seismic behaviors of an existing railway bridge crossing a symmetrical V-shaped canyon.The influence of the source of incident waves is estimated.Numerical results demonstrate that the topographic effects can noticeably amplify the seismic responses of the bridge.Compared to the bridge without crossing a canyon,the peak displacements of the girder,pier,and bearing in the case of the canyon crossing bridge increase by 15.2%,2.9%−14.5%,and 24.2%−229.6%,respectively.The piers at the illuminated side of the canyon experience larger seismic responses compared to the piers at the shaded side of the canyon due to the unequal motion amplitudes at each support.As the source-to-canyon distance increases,the seismic responses of the piers show an increasing trend.展开更多
Deepwater deployment of offshore structures in different sea states was investigated. The whole deployment system was modeled as a lumped mass model, and discretization scheme for cable geometry and methodology for ca...Deepwater deployment of offshore structures in different sea states was investigated. The whole deployment system was modeled as a lumped mass model, and discretization scheme for cable geometry and methodology for calculating the internal and external force acting on deploying cable were presented. The deployment model suitable for the time-varying length of deploying cable was specified. The free-surface flow fields together with the ship motions were used to calculate dynamic tension in the deploying cable during deployment of the structure. The deployment of deep sea mining system which was a typical subsea working system was employed. Based on lumped mass analysis model and parameters of deep sea mining system, numerical simulations were performed, and dynamic load and dynamic amplification factor(DAF) with different cable parameters, deploying velocities and sea states were obtained. It is shown that cable parameters and amplitudes of ocean waves can significantly influence the dynamic load and DAF, and the time-varying natural period of deploying system is a dominant factor, while the effect of deploying velocity is not obvious.展开更多
In order to assess the seismic hazard in an area with high seismogenic potential as the Messina Straits is, we employed the Cornell’s probabilistic approach, implemented by Bender and Perkins in the SEISRISKⅢcode,wh...In order to assess the seismic hazard in an area with high seismogenic potential as the Messina Straits is, we employed the Cornell’s probabilistic approach, implemented by Bender and Perkins in the SEISRISKⅢcode,which allows calculating the expected ground motion.We considered all the earthquakes with magnitude M_s】4.0 reported in the CPTI04 catalogue and located in the seismogenic zones of Southern Italy(ZS9),which are able to展开更多
During the past ten years. our molecular studies haddemonstrated that myelodysplasia of MDS bone marrowwas associated with the rearrangement/amplification ofc-erbB and deletion/ inactivation of c-erbA and identifiedth...During the past ten years. our molecular studies haddemonstrated that myelodysplasia of MDS bone marrowwas associated with the rearrangement/amplification ofc-erbB and deletion/ inactivation of c-erbA and identifiedthe sites of the rearrangement by v-erbB PCR genediagnosis of MDS and confirmed their etiogenicsignificance by the follow-up of MDS cases. These resultsprovided 2 target sequences of c-erbB.The展开更多
The AERORail, a new aerial transport platform, was chosen as the object of this work. Following a review of the literature on static behaviors, model tests on the basic dynamic mechanical characteristics were conducte...The AERORail, a new aerial transport platform, was chosen as the object of this work. Following a review of the literature on static behaviors, model tests on the basic dynamic mechanical characteristics were conducted. A series of 90 tests were completed with different factors, including tension force, vehicle load and vehicle speed. With regard to the proper tension and vehicle load, at a certain speed range, the tension increments of the rail's cable were proved relatively small. It can be assumed that the change of tension is small and can be reasonably ignored when the tension of an entire span is under a dynamic load. When the tension reaches a certain range, the calculation of the cable track structure using classical cable theory is acceptable. The tests prove that the average maximum dynamic amplification factor of the deflection is small, generally no more than 1.2. However, when the vehicle speed reaches a certain value, the amplified factor will reach 2.0. If the moving loads increase, the dynamic amplification factor of dynamic deflection will also increase. The tension will change the rigidity of the structure and the vibration frequency; furthermore, the resonance speed will change at a certain tension. The vibration is noticeable when vehicles pass through at the resonance speed, and this negative impact on driving comfort requires the right velocity to avoid the resonance. The results demonstrate that more design details are required for the AERORail structure.展开更多
The purpose of this study was to establish a method for the rapid detection of infectious pancreatic necrosis virus(IPNV,Jasper serotype)using reverse transcription loop-mediated isothermal amplification(RT-LAMP).Four...The purpose of this study was to establish a method for the rapid detection of infectious pancreatic necrosis virus(IPNV,Jasper serotype)using reverse transcription loop-mediated isothermal amplification(RT-LAMP).Four groups of specific primers were designed,according to the genome sequence of a Chinese IPNV isolate ChRtm213.The results showed that primer set B2 had the best amplification effect.When the final concentration of Mg2+was 6 mmol·L-1,dNTPs were 1 mmol·L-1 and betaine was 0.4 mol·L-1,the reaction could be completed in a 63℃water bath within 60 min.This RT-LAMP assay for the detection of IPNV had no cross-reactivity with infectious hematopoietic necrosis virus,viral hemorrhagic septicemia virus,grass carp reovirus and spring viremia of carp virus.The detection limit was 3.2×10-12 ng·μL-1.The sensitivity of this method was 10-fold higher than that of a previously published RT-LAMP assay for detecting the Spajarup(Sp)serotype of IPNV.This method,aimed at detecting IPNV isolates that were currently prevalent in China,possessed the characteristics of strong specificity,high sensitivity and direct interpretation by the naked eyes.The IPNV RT-LAMP was successfully applied to determine the clinical samples,which indicated the IPNV RT-LAMP assay was suitable for the rapid and large-scale detections of IPNV in China.展开更多
基金the Experimental Technology Research Project of Zhejiang University(SYB202138)National Natural Science Foundation of China(32000195)。
文摘With the approval of more and more genetically modified(GM)crops in our country,GM safety management has become more important.Transgenic detection is a major approach for transgenic safety management.Nevertheless,a convenient and visual technique with low equipment requirements and high sensitivity for the field detection of GM plants is still lacking.On the basis of the existing recombinase polymerase amplification(RPA)technique,we developed a multiplex RPA(multi-RPA)method that can simultaneously detect three transgenic elements,including the cauliflower mosaic virus 35S gene(CaMV35S)promoter,neomycin phosphotransferaseⅡgene(NptⅡ)and hygromycin B phosphotransferase gene(Hyg),thus improving the detection rate.Moreover,we coupled this multi-RPA technique with the CRISPR/Cas12a reporter system,which enabled the detection results to be clearly observed by naked eyes under ultraviolet(UV)light(254 nm;which could be achieved by a portable UV flashlight),therefore establishing a multi-RPA visual detection technique.Compared with the traditional test strip detection method,this multi-RPA-CRISPR/Cas12a technique has the higher specificity,higher sensitivity,wider application range and lower cost.Compared with other polymerase chain reaction(PCR)techniques,it also has the advantages of low equipment requirements and visualization,making it a potentially feasible method for the field detection of GM plants.
基金Project(2014GK3150)supported by Science and Technology Plan of Hunan Province,China
文摘The three-dimensional internal flow field of centrifugal pump is complex and variable with design parameters and operation conditions. The post-processing technique named differential amplification method was proposed for the comparison study of different flow structures. The full steady flow fields of an industrial centrifugal pump working on-design and off-design points were numerically investigated by solving Reynolds average Navier-Stokes equations together with a shear-stress transport(SST) k-? turbulence model. And the numerically predicted performance curves of the studied pump agree well with test measurement results. Compared with the flow flied on design point under the help of differential amplification method, the disturbance caused by interaction between blade and volute tongue is very obvious and it extends to the diffuser pipe on the working point with 0.8 times rated flux. While on the point with 1.2 times rated flux, the flow distribution in impeller region is roughly even and it flows more to the bottom section of the diffuser pipe. The above method was proved to be good at displaying the subtle secondary flow structure changes with a higher resolution effect relative to single isolated case observation, which helps the optimization decision-making from multiple design cases.
基金Project(51278043)supported by National Natural Science Foundation of China
文摘The phenomenon of ground vibration amplification caused by railway traffic was found and proved. In order to study the reasons which cause the amplification, a drop-weight test was performed. Then, the model for both homogeneous and layered soil subjected to a harmonic vertical load was built. With the help of this model, displacement Green's function was calculated and the propagation laws of ground vibration responses were discussed. Results show that: 1) When applying a harmonic load on the half-space surface, the amplitude of ground vibrations attenuate with fluctuation, which is caused by the superposition of bulk and Rayleigh waves. 2) Vibration amplification can be enlarged under the conditions of embedded source and the soil layers. 3) In practice, the fluctuant attenuation should be paid attention to especially for the vibration receivers who are sensitive to single low frequencies(<10 Hz). Moreover, for the case of embedded loads, it should also be paid attention to that the receivers are located at the place where the horizontal distance is similar to embedded depth, usually 10 to 30 m for metro lines.
文摘In order to explore whether the rearrangement and amplification of C-erbB-l gene were associatedwith the development of MDS and their clinicalsignificance in diagnoses and prognoses. digitoxin-labelled oligonucleotide probes were used to detect
文摘When the loop-mediated isothermal amplification(LAMP)assay is used for detecting target genes,DNA extraction is unnecessary in many cases.Simple pretreatment(e.g.heating)is enough to obtain rather sensitive responses.Even test samples without any pretreatment can be used as template.This feature suggests that LAMP is superior to PCR in developing point-of-care test strategies.In this study,using Stx1 gene from E.coli as model,we verified that viable cells,dead cells and extracellular DNA could function as template in the LAMP assay.In the incubation at 63℃,viable bacteria in the LAMP reaction mixture lysed completely within 2 min,providing DNA template for nucleic acid amplification.The Stx1 gene in diluted culture medium,spiked tap water,spiked seawater and real seawater all could be detected,with or without the step of DNA extraction.We found that the complex substances in real sample(e.g.natural seawater)exhibited considerable inhibitory effect on the sensitivity of the LAMP assay.These outcomes are meaningful for building a point-of-care strategy by employing the LAMP assay for environmental monitoring,bio-resource surveys,food safety,etc.in particular those based on environmental DNA.
基金Supported by the Natural Science Foundation of Heilongjiang Province(Topic C2017032)Heilongjiang Province Applied Technology Research and Development Program(Topic GA19B104)the National Key Research and Development Program(Topic 2018YFD0300105)。
文摘Rice bacterial leaf brown spot disease caused by Pseudomonas syringae pv.syringae(Pss)is a major disease on rice.In recent years,Pss has emerged worldwide,seriously affecting rice production.It is very important to establish a rapid detection method of Pss for the diagnosis and prevention of this disease.In order to robust and accurately diagnose the rice bacterial leaf brown spot disease in the field and laboratory,an assay system for the Pss was developed in this study,and the specific sequence of hrcN was used as the target,based on loop-mediated isothermal amplification(LAMP).The best detection system was MgSO 48 mmol·L^(-1),Bst DNA polymerase 8 U,dNTP 1.4 mmol·L^(-1),the ratio of internal and outer primers was 2:1,the reaction temperature was 63℃,the reaction time was 45 min,and the lowest sensitivity was 104 CFU·mL^(-1).This results provided an accurate and robust method for laboratory and field diagnosis of bacterial leaf brown spot disease of rice.
文摘The availability of the plastid genome sequences is one of the bases for comparative,functional,and structural genomic studies of plastid-containing living organisms,in addition to the application
基金the Experimental Technology Research Project of Zhejiang University(SYB202138)National Natural Science Foundation of China(32000195).
文摘Soil DNA extraction,such as microbial community analysis and gene drift detection,is an important basis for multiple analyses in different fields.Nevertheless,the soil DNA extraction methods for field detection are still lacking.This study established a rapid soil DNA extraction(RSDE)method that can be used in field detection.In this method,we first utilized the optimized lysate to isolate DNA from soil and then used a filtration membrane and a DNA adsorption membrane to purify the DNA via the column method.Moreover,we used the pressure from the syringe instead of the conventional centrifugal force of the centrifuge to assist the sample filtration,resulting in very low requirements for this method,with an extraction time of less than 20 min.Furthermore,we demonstrated that the RSDE method was applicable for DNA extraction from different types of soils,with the demand for soil samples as low as 0.1 g and that the amount of obtained DNA was,to some extent,greater than that obtained by a commercial kit.Further analysis revealed that this extracted genomic DNA can be used directly for polymerase chain reaction(PCR)analysis,including ordinary PCR,real-time fluorescent quantitative PCR,and recombinase polymerase amplification(RPA)-CRISPR/Cas12a visual assays.In addition,we demonstrated that this method can be used to extract DNA from residual plant roots in addition to soil microbes,which lays a foundation for the comprehensive analysis of soil plants and microorganisms.In summary,the RSDE method proposed in this study may have wide application prospects.
基金Project(HNTY2022K03)supported by the Hunan Tieyuan Civil Engineering Testing Co.,Ltd.,ChinaProject(52478573)supported by the National Natural Science Foundation of China。
文摘The seismic damage to ancillary facilities on high-speed railway(HSR)bridges can affect the normal movement of trains.To propose the bridge deck acceleration response spectra of the typical HSR simply-supported girder bridge for simplifying the seismic responses analysis of the facilities on bridges,the finite element models of the HSR multi-span simply-supported girder bridges with CRTSII track were established,and the numerical model was validated by tests.Besides,the effects of the span number,peak ground acceleration(PGA),pier height on the seismic acceleration and response spectra of the bridge deck were investigated.Afterward,the bridge acceleration amplification factor curves and bridge deck response spectra with different PGAs and pier heights were obtained.The formula for bridge deck acceleration amplification factor,with a 95%guarantee rate,was fitted.Moreover,the finite element models of the overhead contact lines(OCL)mounted on rigid base and bridges were established to validate the fitted formula.The results indicated that the maximum seismic acceleration response is in the midspan of the beam.The proposed formula for the bridge deck acceleration response spectra can be used to analyze the earthquake response of the OCL and other ancillary facilities on HSR simply-supported girder bridges.The bridge deck acceleration response spectra are conservative in terms of structural safety and can significantly improving the analysis efficiency.
文摘Objective The detection of RNA single nucleotide polymorphism(SNP)is of great importance due to their association with protein expression related to various diseases and drug responses.At present,splintR ligase-assisted methods are important approaches for RNA direct detection,but its specificity will be limited when the fidelity of ligases is not ideal.The aim of this study was to create a method to improve the specificity of splintR ligase for RNA detection.Methods In this study,a dualcompetitive-padlock-probe(DCPLP)assay without the need for additional enzymes or reactions is proposed to improve specificity of splintR ligase ligation.To verify the method,we employed dual competitive padlock probe-mediated rolling circle amplification(DCPLP-RCA)to genotype the CYP2C9 gene.Results The specificity was well improved through the competition and strand displacement of dual padlock probe,with an 83.26%reduction in nonspecific signal.By detecting synthetic RNA samples,the method demonstrated a dynamic detection range of 10 pmol/L-1 nmol/L.Furthermore,clinical samples were applied to the method to evaluate its performance,and the genotyping results were consistent with those obtained using the qPCR method.Conclusion This study has successfully established a highly specific direct RNA SNP detection method,and provided a novel avenue for accurate identification of various types of RNAs.
基金Projects(51908123,51678140,U1934205)supported by the National Natural Science Foundation of ChinaProjects(BK20190370,BK20180383) supported by the Natural Science Foundation of Jiangsu Province,China+1 种基金Project(18KJA580005) supported by the Natural Science Foundation of Jiangsu Higher Education Institutions of ChinaProject(KYY2019096(19-21)) supported by the Science and Technology Research Plan of China Railway Eryuan Engineering Group Corporation。
文摘Past earthquakes have revealed that topographic features have significant impacts on the characteristics of ground motions,which may cause the amplification and de-amplification of input seismic waves.The topographic effect with the assumption of plane seismic waves on the seismic responses of bridges has been investigated in the existing literature;however,the influence of near-source topographic effects has not been thoroughly understood.The objective of this study is to numerically explore the near-source topographic effects on the seismic behaviors of an existing railway bridge crossing a symmetrical V-shaped canyon.The influence of the source of incident waves is estimated.Numerical results demonstrate that the topographic effects can noticeably amplify the seismic responses of the bridge.Compared to the bridge without crossing a canyon,the peak displacements of the girder,pier,and bearing in the case of the canyon crossing bridge increase by 15.2%,2.9%−14.5%,and 24.2%−229.6%,respectively.The piers at the illuminated side of the canyon experience larger seismic responses compared to the piers at the shaded side of the canyon due to the unequal motion amplitudes at each support.As the source-to-canyon distance increases,the seismic responses of the piers show an increasing trend.
基金Project(51305463) supported by the National Natural Science Foundation of China
文摘Deepwater deployment of offshore structures in different sea states was investigated. The whole deployment system was modeled as a lumped mass model, and discretization scheme for cable geometry and methodology for calculating the internal and external force acting on deploying cable were presented. The deployment model suitable for the time-varying length of deploying cable was specified. The free-surface flow fields together with the ship motions were used to calculate dynamic tension in the deploying cable during deployment of the structure. The deployment of deep sea mining system which was a typical subsea working system was employed. Based on lumped mass analysis model and parameters of deep sea mining system, numerical simulations were performed, and dynamic load and dynamic amplification factor(DAF) with different cable parameters, deploying velocities and sea states were obtained. It is shown that cable parameters and amplitudes of ocean waves can significantly influence the dynamic load and DAF, and the time-varying natural period of deploying system is a dominant factor, while the effect of deploying velocity is not obvious.
文摘In order to assess the seismic hazard in an area with high seismogenic potential as the Messina Straits is, we employed the Cornell’s probabilistic approach, implemented by Bender and Perkins in the SEISRISKⅢcode,which allows calculating the expected ground motion.We considered all the earthquakes with magnitude M_s】4.0 reported in the CPTI04 catalogue and located in the seismogenic zones of Southern Italy(ZS9),which are able to
文摘During the past ten years. our molecular studies haddemonstrated that myelodysplasia of MDS bone marrowwas associated with the rearrangement/amplification ofc-erbB and deletion/ inactivation of c-erbA and identifiedthe sites of the rearrangement by v-erbB PCR genediagnosis of MDS and confirmed their etiogenicsignificance by the follow-up of MDS cases. These resultsprovided 2 target sequences of c-erbB.The
基金Projects(50708072,51378385)supported by the National Natural Science Foundation of China
文摘The AERORail, a new aerial transport platform, was chosen as the object of this work. Following a review of the literature on static behaviors, model tests on the basic dynamic mechanical characteristics were conducted. A series of 90 tests were completed with different factors, including tension force, vehicle load and vehicle speed. With regard to the proper tension and vehicle load, at a certain speed range, the tension increments of the rail's cable were proved relatively small. It can be assumed that the change of tension is small and can be reasonably ignored when the tension of an entire span is under a dynamic load. When the tension reaches a certain range, the calculation of the cable track structure using classical cable theory is acceptable. The tests prove that the average maximum dynamic amplification factor of the deflection is small, generally no more than 1.2. However, when the vehicle speed reaches a certain value, the amplified factor will reach 2.0. If the moving loads increase, the dynamic amplification factor of dynamic deflection will also increase. The tension will change the rigidity of the structure and the vibration frequency; furthermore, the resonance speed will change at a certain tension. The vibration is noticeable when vehicles pass through at the resonance speed, and this negative impact on driving comfort requires the right velocity to avoid the resonance. The results demonstrate that more design details are required for the AERORail structure.
基金Supported by the National Natural Science Foundation of China(31802345)China Postdoctoral Science Foundation(2018M630893)Heilongjiang Province Postdoctoral Science Foundation(LBH-Z18275)。
文摘The purpose of this study was to establish a method for the rapid detection of infectious pancreatic necrosis virus(IPNV,Jasper serotype)using reverse transcription loop-mediated isothermal amplification(RT-LAMP).Four groups of specific primers were designed,according to the genome sequence of a Chinese IPNV isolate ChRtm213.The results showed that primer set B2 had the best amplification effect.When the final concentration of Mg2+was 6 mmol·L-1,dNTPs were 1 mmol·L-1 and betaine was 0.4 mol·L-1,the reaction could be completed in a 63℃water bath within 60 min.This RT-LAMP assay for the detection of IPNV had no cross-reactivity with infectious hematopoietic necrosis virus,viral hemorrhagic septicemia virus,grass carp reovirus and spring viremia of carp virus.The detection limit was 3.2×10-12 ng·μL-1.The sensitivity of this method was 10-fold higher than that of a previously published RT-LAMP assay for detecting the Spajarup(Sp)serotype of IPNV.This method,aimed at detecting IPNV isolates that were currently prevalent in China,possessed the characteristics of strong specificity,high sensitivity and direct interpretation by the naked eyes.The IPNV RT-LAMP was successfully applied to determine the clinical samples,which indicated the IPNV RT-LAMP assay was suitable for the rapid and large-scale detections of IPNV in China.
