Three genetic mechanisms activate oncogenes in human neoplasms: 1) mutations, 2) gene amplification, and 3) chromosome rearrangements. These mechanisms result in either an alteration of protooncogene structure or an i...Three genetic mechanisms activate oncogenes in human neoplasms: 1) mutations, 2) gene amplification, and 3) chromosome rearrangements. These mechanisms result in either an alteration of protooncogene structure or an increase in protooncogene expression. The role of epigenetic aberrancies in carcinogenesis has been described earlier however to clinicians, the biological implications of epigenetic therapies to prevent cancer and the mechanisms involved have been a mystery. Furthermore, there is no biomarker suggested to track the carcinogenesis steps long before cancer develops, and this has caused a significant lack of proactive and preventive measures to be taken as all recommendations in preventive oncology are either deficiently and blindly made or through screening methods which are too late in the game. Here we explored a very different approach by applying our deepest understanding of epigenetics and carcinogenesis and even further we developed a framework where our clinical findings could translate to the research and vice versa by generating advanced and novel hypotheses on “how we get cancer”, by exploring the relation between the host and the tumor cells in a way no one had perceived before. The role of specific cancer stem cell pathways is dissected and how to inhibit each of these initiators using multitargeted epigenetic therapies and off-label medications are explained. We should admit that without considering this sophisticated amazing biological network, cancer will remain an unsolved challenge. Further, we were able to solve this unsolved puzzle by bridging the gap from a hypothetical point of view/hypothesis to possibilities that explain the clinical findings we had observed, and conclude that such an approach can completely change the way practitioners are treating cancer.展开更多
The kelp grouper (Epinephelus bruneus), belonging to one of the largest genera among the subfamily Epinephelinae, is a commercially important fish in Japan. There are limited data about the genomics of this species. T...The kelp grouper (Epinephelus bruneus), belonging to one of the largest genera among the subfamily Epinephelinae, is a commercially important fish in Japan. There are limited data about the genomics of this species. To provide tools for addressing both population genetics studies and gene mapping, dito pentanucleotide simple sequence repeat (SSR) markers were developed using 454 pyrosequencing. Among the 1466 SSR markers developed, 1244 primer sets produced strong PCR products, of which 905 (72.7%) were polymorphic in kelp grouper. Cross-species utility of the 905 polymorphic SSR markers was tested in four additional Epinephelinae species of Hyporthodus septemfasciatus, Plectropomus leopardus, Epinephelus lanceolatus and Epinephelus coioides. Results revealed that, respectively, 401 (44.3%), 136 (15.0%), 434 (49.0%) and 538 (59.4%) SSRs showed specific polymorphic products. Of these, 40 SSR markers (33 di-, 1 tri- and 6 tetra-nucleotides) showed polymorphism in all species tested. Additionally, three AGAT SSR motifs which accounted for 42.9% of the nondi-nucleotide markers were found in the 40 SSR markers. This indicates that the AGAT SSR motif has a high potential as a highly versatile SSR marker in grouper Epinephelinae. The SSR markers developed in this study can be employed to obtain reliable genetic variability estimates for groupers (Epinephelinae).展开更多
The theory of slow backward-wave amplifications is developed based on electron cyclotron maser （ECM） mechanism employing an initially rectilinear beam, A nonlinear evolution equation is derived to describe the elect...The theory of slow backward-wave amplifications is developed based on electron cyclotron maser （ECM） mechanism employing an initially rectilinear beam, A nonlinear evolution equation is derived to describe the electron energy. Numerical calculations show that the saturated interaction efficiency in this system may exceed 20~, and the saturated interaction length spans 3-6 centimeters. The distinctive interaction mechanism is promising for the design of compact backward microwave amplification devices, Numerical studies are also presented for the slow-wave ECM efficiency with inclusion of Gaussian beam electron velocity spread. It is shown that the velocity spread reduces the interaction e^ciency.展开更多
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
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
Thirty-two cases of ovarian carcinoma, two of normal ovaries, four of benign epithelial ovarian tumor, and three of borderline epithelial ovarian tumor were studied using Southern blot hybridization of DNA. In 15 of t...Thirty-two cases of ovarian carcinoma, two of normal ovaries, four of benign epithelial ovarian tumor, and three of borderline epithelial ovarian tumor were studied using Southern blot hybridization of DNA. In 15 of the 32 cases of ovarian carcinoma, peripheral lymphocytes were also studied. The amplification rate of C-myc, C-N-ras, C-Ki-ras and C-erbB-2 in ovarian carcinoma were 50%, 44%, 31% and 25% respectively. The amplification of C-Ki-ras and C-N-ras took place chiefly in cases of early stage and those of good differentiation. The amplification of C-N-ras was also found in cases of advanced stage. The amplifications of C-myc and C-erbB-2 were chiefly found in cases above stage Ⅲ and those of poor differentiation. A total of 83% of the patients who died were found to have amplifications of more than 2 proto-oncogenes, with which the amplification of C-erbB-2 was involved.展开更多
We compare different discreted DCF Raman amplifier configurations, including single-stage and dual-stage. The optimum design with respect to SNR degradation, compromise linear and nonlinear impairments.
The seismic design forces of nonstructural components(NSCs)in buildings are closely related to floor acceleration response amplification.To investigate the differences in acceleration responses of structures with diff...The seismic design forces of nonstructural components(NSCs)in buildings are closely related to floor acceleration response amplification.To investigate the differences in acceleration responses of structures with different structural types,fundamental periods,and seismic design levels,56 reinforced concrete and steel structures with fundamental periods ranging from 0.37 s to 5.68 s were selected.For each structure,100 sets of earthquake motions were used as inputs for elastic time history analysis.Based on the resulting 26,500 sets of floor acceleration response data,the amplification rules of peak floor acceleration/peak ground acceleration(PFA/PGA)along the height of various structures and the corresponding floor response spectrum characteristics were studied.The nonlinear changes of PFA/PGA along the height of long period structures were compared with the codes of different countries.Moreover,more suitable prediction equations were proposed based on the structural characteristics.Finally,to solve the issue that existing research still cannot accurately reflect the acceleration amplification coefficient of NSCs with different dynamic characteristics in main structures with different periods,a normalized floor response spectrum is proposed that can simultaneously consider the effects of input ground motion characteristics and the main structure,which can be better used in the seismic design of NSCs.展开更多
Strand displacement-based DNA circuits have emerged as highly effective tools for molecular computation,serving purposes of amplification or decision-making.They are favored for their inherent occurrence and sensitivi...Strand displacement-based DNA circuits have emerged as highly effective tools for molecular computation,serving purposes of amplification or decision-making.They are favored for their inherent occurrence and sensitivity to external conditions.However,achieving enhanced amplification or decision-making necessitates the incorporation of multiple strands,thereby increasing the risk of contamination.Recent advancements have led to the development of CRISPR-Cas-based DNA circuits.These systems aim to simplify the complexity associated with conventional circuits,mitigate contamination risks,and enable more substantial amplification or decision-making capabilities.Here,the review article centers on current strategies of CRISPR-Cas(Cas9,Cas12a,Cas13a)system-assisted circuits in amplification and decisionmaking,and assesses their tendencies and limitations in amplification circuits and decision-making circuits.Furthermore,we discuss the challenges of CRISPR-Cas in circuits and propose prospects that will contribute to constructing more efficient and diverse CRISPR-Cas-based DNA functional circuits.展开更多
Glial fibrillary acidic protein(GFAP)is one of the discriminative biomarkers for diagnosing traumatic brain injury(TBI),and accurate determination of GFAP is clinically significant.In this study,a novel fluorescence i...Glial fibrillary acidic protein(GFAP)is one of the discriminative biomarkers for diagnosing traumatic brain injury(TBI),and accurate determination of GFAP is clinically significant.In this study,a novel fluorescence immunoassay system was designed.We encapsulated carbon dots with a high fluorescence quantum yield(QY=92.5%)inside silicon nanocapsules to serve as fluorescent markers.These markers were then integrated with the streptavidin(SA)-biotin biomagnification system and immunomagnetic separation technology for the sensitive detection of GFAP.Based on the signal cascade amplification effect of the silicon nanocapsules and SA-biotin,the fluorescence signal of the SA-biotin-modified immunofluorescence nanocapsules increased 3.6-fold compared to the carbon dot-based immunoprobe.The fluorescence immunoassay system was constructed for GFAP using SA-biotin-modified immunocapsules as the sensing probe and immunomagnetic nanoparticles as the immunorecognition probe.The fluorescence immunoassay system can specifically and ultra-sensitively quantify GFAP in blood samples,with a detection range of 10 pg/mL–10 ng/mL and detection limits of 3.2 pg/mL(serum)and 3.6 pg/mL(plasma).Moreover,the fluorescence immunoassay system exhibited prominent recoveries of 99.4%–100.4%(phosphate buffered saline),96%–102.6%(serum),and 93.2%–110.2%(plasma),with favorable specificity and excellent stabilization.The novel fluorescence immunoassay system provides a new approach to the clinical analysis of GFAP and may serve as a potential tool for screening and diagnosing TBI.展开更多
Lateral flow immunoassay(LFIA),a rapid detection technique noted for simplicity and economy,has showcased indispensable applicability in diverse domains such as disease screening,food safety,and environmental monitori...Lateral flow immunoassay(LFIA),a rapid detection technique noted for simplicity and economy,has showcased indispensable applicability in diverse domains such as disease screening,food safety,and environmental monitoring.Nevertheless,challenges still exist in detecting ultra-low concentration analytes due to the inherent sensitivity limitations of LFIA.Recently,significant advances have been achieved by integrating enzyme activity probes and transforming LFIA into a highly sensitive tool for rapidly detecting trace analyte concentrations.Specifically,modifying natural enzymes or engineered nanozymes allows them to function as immune probes,directly catalyzing the production of signal molecules or indirectly initiating enzyme activity.Therefore,the signal intensity and detection sensitivity of LFIA are markedly elevated.The present review undertakes a comprehensive examination of pertinent research literature,offering a systematic analysis of recently proposed enzyme-based signal amplification strategies.By way of comparative assessment,the merits and demerits of current approaches are delineated,along with the identification of research avenues that still need to be explored.It is anticipated that this critical overview will garner considerable attention within the biomedical and materials science communities,providing valuable direction and insight toward the advancement of high-performance LFIA technologies.展开更多
The purpose of this article is to analyze the efficacy of molecular diagnostic techniques for monitoring tuberculosis relapse.After analyzing the connotation of molecular diagnostic techniques and their specific appli...The purpose of this article is to analyze the efficacy of molecular diagnostic techniques for monitoring tuberculosis relapse.After analyzing the connotation of molecular diagnostic techniques and their specific application process in tuberculosis relapse monitoring,a total of 200 cured tuberculosis patients were selected(100 in the experimental group and 100 in the control group).During the 12-month follow-up period,the experimental group was monitored by molecular diagnostic techniques,while the control group was monitored by traditional techniques.Finally,by comparing the performance indicators of the two monitoring methods,as well as the relapse situations and outcomes of patients,it was demonstrated that molecular diagnostic techniques have higher efficacy in tuberculosis relapse monitoring.展开更多
Ultrasensitive detection of nucleic acids is of great significance for precision medicine.Digital polymerase chain reaction(dPCR)is the most sensitive method but requires sophisticated and expensive instruments and a ...Ultrasensitive detection of nucleic acids is of great significance for precision medicine.Digital polymerase chain reaction(dPCR)is the most sensitive method but requires sophisticated and expensive instruments and a long reaction time.Digital PCR-free technologies,which mean the digital assay not relying on thermal cycling to amplify the signal for quantitative detection of nucleic acids at the singlemolecule level,include the digital isothermal amplification techniques(d IATs)and the digital clustered regularly interspaced short palindromic repeats(CRISPR)technologies.They combine the advantages of d PCR and IATs,which could be fast and simple,enabling absolute quantification of nucleic acids at a single-molecule level with minimum instrument,representing the next-generation molecular diagnostic technology.Herein,we systematically summarized the strategies and applications of various dIATs,including the digital loop-mediated isothermal amplification(dLAMP),the digital recombinase polymerase amplification(dRPA),the digital rolling circle amplification(dRCA),the digital nucleic acid sequencebased amplification(d NASBA)and the digital multiple displacement amplification(d MDA),and evaluated the pros and cons of each method.The emerging digital CRISPR technologies,including the detection mechanism of CRISPR and the various strategies for signal amplification,are also introduced comprehensively in this review.The current challenges as well as the future perspectives of the digital PCR-free technology were discussed.展开更多
[Objectives]To develop a pair of specific primers for the PCR amplification of the full-length relA gene from Vibrio alginolyticus strain HY9901,as well as to conduct bioinformatics analysis.[Methods]The relA gene was...[Objectives]To develop a pair of specific primers for the PCR amplification of the full-length relA gene from Vibrio alginolyticus strain HY9901,as well as to conduct bioinformatics analysis.[Methods]The relA gene was amplified through PCR,and the resulting gene sequence was subsequently analyzed using bioinformatics tools,including amino acid sequence prediction,functional site analysis,subcellular localization prediction,and homology comparison.[Results]The relA gene had a total length of 2220 bp and encoded 739 amino acid residues.The molecular weight was approximately 84.1261 kDa,and its isoelectric point was 5.95.The protein lacked a signal peptide and transmembrane regions,while exhibiting multiple phosphorylation sites.Predictions regarding its subcellular localization suggested that it was predominantly situated in the cytoplasm.The amino acid sequence demonstrated a homology of 97%to 99%with other species within the genus Vibrio,and it clustered within the same subfamily as V.antiquarius and V.diabolicus.In the prediction of secondary structure,the proportions ofα-helix,extended strand,random coil,andβ-sheet were 54.13%,12.04%,28.15%and 5.68%,respectively.The similarity between the tertiary structure model and template 5kpw.1.w was 66%.[Conclusions]In this study,the relA gene of V.alginolyticus strain HY9901 has been successfully amplified and analyzed.The structural characteristics and potential functions of the encoded protein have been elucidated,thereby providing foundational data for understanding the role of this gene in V.alginolyticus.展开更多
Although diverse signal-amplified methods have been committed to improve the sensitivity of surface plasmon resonance(SPR)biosensing,introducing convenient and robust signal amplification strategy into SPR biosensing ...Although diverse signal-amplified methods have been committed to improve the sensitivity of surface plasmon resonance(SPR)biosensing,introducing convenient and robust signal amplification strategy into SPR biosensing remains challenging.Here,a novel nanozyme-triggered polymerization amplification strategy was proposed for constructing highly sensitive surface plasmon resonance(SPR)immunosensor.In detail,Au@Pd core-shell nanooctahedra nanozyme with superior peroxidase(POD)-like activity was synthesized and utilized as a label probe.Simultaneously,Au@Pd core-shell nanooctahedra nanozyme can catalyze the decomposition of H_(2)O_(2)to form hydroxyl radicals(·OH)that triggers the polymerization of aniline to form polyaniline attaching on the surface of sensor chip,significantly amplifying SPR responses.The sensitivity of SPR immunosensor was enhanced by nanozyme-triggered polymerization amplification strategy.Using human immunoglobulin G(HIgG)as a model,the constructed SPR immunosensor obtains a wide linear range of 0.005–1.0μg/m L with low detection limit of 0.106 ng/m L.This research provides new sights on establishing sensitive SPR immunosensor and may evokes more inspiration for developing signal amplification methods based on nanozyme in biosensing.展开更多
To capture the nonlinear dynamics and gain evolution in chirped pulse amplification(CPA)systems,the split-step Fourier method and the fourth-order Runge–Kutta method are integrated to iteratively address the generali...To capture the nonlinear dynamics and gain evolution in chirped pulse amplification(CPA)systems,the split-step Fourier method and the fourth-order Runge–Kutta method are integrated to iteratively address the generalized nonlinear Schrödinger equation and the rate equations.However,this approach is burdened by substantial computational demands,resulting in significant time expenditures.In the context of intelligent laser optimization and inverse design,the necessity for numerous simulations further exacerbates this issue,highlighting the need for fast and accurate simulation methodologies.Here,we introduce an end-to-end model augmented with active learning(E2E-AL)with decent generalization through different dedicated embedding methods over various parameters.On an identical computational platform,the artificial intelligence–driven model is 2000 times faster than the conventional simulation method.Benefiting from the active learning strategy,the E2E-AL model achieves decent precision with only two-thirds of the training samples compared with the case without such a strategy.Furthermore,we demonstrate a multi-objective inverse design of the CPA systems enabled by the E2E-AL model.The E2E-AL framework manifests the potential of becoming a standard approach for the rapid and accurate modeling of ultrafast lasers and is readily extended to simulate other complex systems.展开更多
DNA repair enzymes are important in the repair of DNA lesions for maintaining the genome stability,and their abnormal expression induced various human cancers.Simultaneous detection of these DNA enzymes could provide ...DNA repair enzymes are important in the repair of DNA lesions for maintaining the genome stability,and their abnormal expression induced various human cancers.Simultaneous detection of these DNA enzymes could provide convincing evidence based on the comparison of the activity of multiple enzymes than on that of single enzyme.Although fluorescence approach has been applied for the simultaneous detection both of DNA repair enzymes,the spectral overlap and multiwavelength excitation severely restrict the number of available fluorophores.Thus,it is difficult to simultaneously detect three enzymes in a single analysis by fluorescence detection.Herein,we developed a method for the simultaneous determination of three DNA repair enzymes including human flap DNA endonuclease 1(FEN1),human alkyladenine DNA glycosylase(hAAG)and uracil DNA glycosylase(UDG)based on the combination of template-free amplification system with capillary electrophoresis-laser induced fluorescence(CE-LIF)detection.The amplification system was adopted to transfer and amplify the enzymatic products into different length DNA fragments which could be separated effectively by CE-LIF without the complicated modification of the capillary inner wall or labeling different tails on signal probes for separation.The method demonstrated a detection limit of 0.07 U/mL(0.08-160 U/mL)for FEN1,2.40 U/mL(2.5-250U/mL)for hAAG and 2.1×10^(-4)U/mL(0.0004-2.5 U/mL)for UDG,the relative standard deviations(RSDs)of peak time and peak area for different analytes were as follows:2.50%-4,37%and 3.24%-7.18%(inter-day);1.37%-2.71%and 1.43%-3.02%(intra-day),4.28%-6.08%and 4.16%-7.57%(column to column),respectively.And it can identify the inhibitor-like drugs,evaluate enzymatic kinetics and achieve the detection of three enzymes in cell extracts,providing a simple and powerful platform for simultaneous detection of more DNA repair enzymes.展开更多
Amplification-free,highly sensitive,and specific nucleic acid detection is crucial for health monitoring and diagnosis.The type III CRISPR-Cas10 system,which provides viral immunity through CRISPRassociated protein ef...Amplification-free,highly sensitive,and specific nucleic acid detection is crucial for health monitoring and diagnosis.The type III CRISPR-Cas10 system,which provides viral immunity through CRISPRassociated protein effectors,enables a new amplification-free nucleic acid diagnostic tool.In this study,we develop a CRISPR-graphene field-effect transistors(GFETs)biosensor by combining the type III CRISPR-Cas10 system with GFETs for direct nucleic acid detection.This biosensor exploits the target RNA-activated continuous ss DNA cleavage activity of the d Csm3 CRISPR-Cas10 effector and the high charge density of a hairpin DNA reporter on the GFET channel to achieve label-free,amplification-free,highly sensitive,and specific RNA detection.The CRISPR-GFET biosensor exhibits excellent performance in detecting medium-length RNAs and miRNAs,with detection limits at the aM level and a broad linear range of 10^(-15)to 10^(-11)M for RNAs and 10^(-15)to 10^(-9)M for miRNAs.It shows high sensitivity in throat swabs and serum samples,distinguishing between healthy individuals(N=5)and breast cancer patients(N=6)without the need for extraction,purification,or amplification.This platform mitigates risks associated with nucleic acid amplification and cross-contamination,making it a versatile and scalable diagnostic tool for molecular diagnostics in human health.展开更多
Bacterial blight(BB) is a devastating worldwide rice disease caused by Xanthomonas oryzae pv. oryzae(Xoo), which is difficult to diagnose based on early symptoms. Conventional chemical control yields limited effective...Bacterial blight(BB) is a devastating worldwide rice disease caused by Xanthomonas oryzae pv. oryzae(Xoo), which is difficult to diagnose based on early symptoms. Conventional chemical control yields limited effectiveness once BB has spread. Consequently, it is imperative to develop a rapid, highly sensitive, specific, and easy-to-use detection technique for early on-site diagnosis of BB. We first developed a recombinase-aided amplification-lateral flow dipstick(RAA-LFD) technique for the on-site detection of Xoo. The optimized reaction temperature and time were 37 ℃ and 20 min, indicating that the reaction system can be initiated by body temperature independently of any precision instruments. Evaluation of the RAA-LFD technique using the primers(RAAF2/R2) and probe(RAA2-nfo-probe) derived from the Xoo ORF0080 locus exhibited high specificity and eliminated cross-reactivity with other bacterial species. The sensitivity of RAA-LFD is up to 1 pg/μL for Xoo genomic DNA and 100 CFU/m L for Xoo cells. Significantly, this technique accurately detected Xoo from both artificially inoculated and naturally infected rice leaves at the early stage of infection, directly deploying plant tissue fluid as the template without DNA extraction. These attributes make the developed RAA-LFD system a viable technique for the early diagnosis of BB in the field, providing technical support for early-warning systems and disease control.展开更多
Development of accurate analytical protocols for cancer biomarkers is used for the initial prescreening of malignant tumors,disease surveillance,and efficacy assessment with significant clinical benefits.In this work,...Development of accurate analytical protocols for cancer biomarkers is used for the initial prescreening of malignant tumors,disease surveillance,and efficacy assessment with significant clinical benefits.In this work,we reported a liposome-mediated signal-off photoelectrochemical(PEC)immunoassay for the sensitive detection of carcinoembryonic antigen(CEA)using ternary transition metal sulfide CuS/ZnCdS as the photoactive material.Good photocurrents were acquired on the basis of specific oxidation reaction of dopamine on the CuS/ZnCdS.The energy band relationship of CuS/ZnCdS was determined,and the wellmatched oxidation potential of dopamine was verified.To achieve accurate recovery of low-abundance CEA,systematic PEC evaluation from human serum samples was performed by combining with classical immunoreaction and liposome-induced dopamine amplification strategy with high stability and selectivity.Under optimum conditions,PEC immunoassay displayed good photocurrent responses toward target CEA with a dynamic linear range of 0.1-50 ng/mL with a detection limit of 31.6 pg/mL.Importantly,this system by combining with a discussion of energy level matching between semiconductor energy bands and small-molecules opens a new horizon for development of high-efficient PEC immunoassays.展开更多
文摘Three genetic mechanisms activate oncogenes in human neoplasms: 1) mutations, 2) gene amplification, and 3) chromosome rearrangements. These mechanisms result in either an alteration of protooncogene structure or an increase in protooncogene expression. The role of epigenetic aberrancies in carcinogenesis has been described earlier however to clinicians, the biological implications of epigenetic therapies to prevent cancer and the mechanisms involved have been a mystery. Furthermore, there is no biomarker suggested to track the carcinogenesis steps long before cancer develops, and this has caused a significant lack of proactive and preventive measures to be taken as all recommendations in preventive oncology are either deficiently and blindly made or through screening methods which are too late in the game. Here we explored a very different approach by applying our deepest understanding of epigenetics and carcinogenesis and even further we developed a framework where our clinical findings could translate to the research and vice versa by generating advanced and novel hypotheses on “how we get cancer”, by exploring the relation between the host and the tumor cells in a way no one had perceived before. The role of specific cancer stem cell pathways is dissected and how to inhibit each of these initiators using multitargeted epigenetic therapies and off-label medications are explained. We should admit that without considering this sophisticated amazing biological network, cancer will remain an unsolved challenge. Further, we were able to solve this unsolved puzzle by bridging the gap from a hypothetical point of view/hypothesis to possibilities that explain the clinical findings we had observed, and conclude that such an approach can completely change the way practitioners are treating cancer.
文摘The kelp grouper (Epinephelus bruneus), belonging to one of the largest genera among the subfamily Epinephelinae, is a commercially important fish in Japan. There are limited data about the genomics of this species. To provide tools for addressing both population genetics studies and gene mapping, dito pentanucleotide simple sequence repeat (SSR) markers were developed using 454 pyrosequencing. Among the 1466 SSR markers developed, 1244 primer sets produced strong PCR products, of which 905 (72.7%) were polymorphic in kelp grouper. Cross-species utility of the 905 polymorphic SSR markers was tested in four additional Epinephelinae species of Hyporthodus septemfasciatus, Plectropomus leopardus, Epinephelus lanceolatus and Epinephelus coioides. Results revealed that, respectively, 401 (44.3%), 136 (15.0%), 434 (49.0%) and 538 (59.4%) SSRs showed specific polymorphic products. Of these, 40 SSR markers (33 di-, 1 tri- and 6 tetra-nucleotides) showed polymorphism in all species tested. Additionally, three AGAT SSR motifs which accounted for 42.9% of the nondi-nucleotide markers were found in the 40 SSR markers. This indicates that the AGAT SSR motif has a high potential as a highly versatile SSR marker in grouper Epinephelinae. The SSR markers developed in this study can be employed to obtain reliable genetic variability estimates for groupers (Epinephelinae).
基金supported by National Natural Science Foundation of China(Nos.11275007 and 11175023)the Program for Liaoning Excellent Talents in University(LJQ2012098)
文摘The theory of slow backward-wave amplifications is developed based on electron cyclotron maser （ECM） mechanism employing an initially rectilinear beam, A nonlinear evolution equation is derived to describe the electron energy. Numerical calculations show that the saturated interaction efficiency in this system may exceed 20~, and the saturated interaction length spans 3-6 centimeters. The distinctive interaction mechanism is promising for the design of compact backward microwave amplification devices, Numerical studies are also presented for the slow-wave ECM efficiency with inclusion of Gaussian beam electron velocity spread. It is shown that the velocity spread reduces the interaction e^ciency.
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.
文摘Thirty-two cases of ovarian carcinoma, two of normal ovaries, four of benign epithelial ovarian tumor, and three of borderline epithelial ovarian tumor were studied using Southern blot hybridization of DNA. In 15 of the 32 cases of ovarian carcinoma, peripheral lymphocytes were also studied. The amplification rate of C-myc, C-N-ras, C-Ki-ras and C-erbB-2 in ovarian carcinoma were 50%, 44%, 31% and 25% respectively. The amplification of C-Ki-ras and C-N-ras took place chiefly in cases of early stage and those of good differentiation. The amplification of C-N-ras was also found in cases of advanced stage. The amplifications of C-myc and C-erbB-2 were chiefly found in cases above stage Ⅲ and those of poor differentiation. A total of 83% of the patients who died were found to have amplifications of more than 2 proto-oncogenes, with which the amplification of C-erbB-2 was involved.
文摘We compare different discreted DCF Raman amplifier configurations, including single-stage and dual-stage. The optimum design with respect to SNR degradation, compromise linear and nonlinear impairments.
基金Natural Science Foundation of China under Grant Nos.52078471,52078472 and 52208509National Key Research and Development Plan of China under Grant No.2019YFE0112700+2 种基金Natural Science Foundation of Heilongjiang Province under Grant No.LH2022E121Special Project for Basic Scientific Research Business Expenses of the Institute of Engineering Mechanics,China Earthquake Administration under Grant No.2022C04Director’s Fund Director’s Fund of Earthquake Agency of Inner Mongolia Autonomous Region under Grant No.2023MS10。
文摘The seismic design forces of nonstructural components(NSCs)in buildings are closely related to floor acceleration response amplification.To investigate the differences in acceleration responses of structures with different structural types,fundamental periods,and seismic design levels,56 reinforced concrete and steel structures with fundamental periods ranging from 0.37 s to 5.68 s were selected.For each structure,100 sets of earthquake motions were used as inputs for elastic time history analysis.Based on the resulting 26,500 sets of floor acceleration response data,the amplification rules of peak floor acceleration/peak ground acceleration(PFA/PGA)along the height of various structures and the corresponding floor response spectrum characteristics were studied.The nonlinear changes of PFA/PGA along the height of long period structures were compared with the codes of different countries.Moreover,more suitable prediction equations were proposed based on the structural characteristics.Finally,to solve the issue that existing research still cannot accurately reflect the acceleration amplification coefficient of NSCs with different dynamic characteristics in main structures with different periods,a normalized floor response spectrum is proposed that can simultaneously consider the effects of input ground motion characteristics and the main structure,which can be better used in the seismic design of NSCs.
基金financially supported by the National Natural Science Foundation of China (Nos. 82172372 and 82260290)the Opening Research Fund of State Key Laboratory of Digital Medical Engineering (No. 2023-M04)
文摘Strand displacement-based DNA circuits have emerged as highly effective tools for molecular computation,serving purposes of amplification or decision-making.They are favored for their inherent occurrence and sensitivity to external conditions.However,achieving enhanced amplification or decision-making necessitates the incorporation of multiple strands,thereby increasing the risk of contamination.Recent advancements have led to the development of CRISPR-Cas-based DNA circuits.These systems aim to simplify the complexity associated with conventional circuits,mitigate contamination risks,and enable more substantial amplification or decision-making capabilities.Here,the review article centers on current strategies of CRISPR-Cas(Cas9,Cas12a,Cas13a)system-assisted circuits in amplification and decisionmaking,and assesses their tendencies and limitations in amplification circuits and decision-making circuits.Furthermore,we discuss the challenges of CRISPR-Cas in circuits and propose prospects that will contribute to constructing more efficient and diverse CRISPR-Cas-based DNA functional circuits.
基金supported by the AMS Funding Project(No.ZZB2023C7010).
文摘Glial fibrillary acidic protein(GFAP)is one of the discriminative biomarkers for diagnosing traumatic brain injury(TBI),and accurate determination of GFAP is clinically significant.In this study,a novel fluorescence immunoassay system was designed.We encapsulated carbon dots with a high fluorescence quantum yield(QY=92.5%)inside silicon nanocapsules to serve as fluorescent markers.These markers were then integrated with the streptavidin(SA)-biotin biomagnification system and immunomagnetic separation technology for the sensitive detection of GFAP.Based on the signal cascade amplification effect of the silicon nanocapsules and SA-biotin,the fluorescence signal of the SA-biotin-modified immunofluorescence nanocapsules increased 3.6-fold compared to the carbon dot-based immunoprobe.The fluorescence immunoassay system was constructed for GFAP using SA-biotin-modified immunocapsules as the sensing probe and immunomagnetic nanoparticles as the immunorecognition probe.The fluorescence immunoassay system can specifically and ultra-sensitively quantify GFAP in blood samples,with a detection range of 10 pg/mL–10 ng/mL and detection limits of 3.2 pg/mL(serum)and 3.6 pg/mL(plasma).Moreover,the fluorescence immunoassay system exhibited prominent recoveries of 99.4%–100.4%(phosphate buffered saline),96%–102.6%(serum),and 93.2%–110.2%(plasma),with favorable specificity and excellent stabilization.The novel fluorescence immunoassay system provides a new approach to the clinical analysis of GFAP and may serve as a potential tool for screening and diagnosing TBI.
基金Financial supports from the National Natural Science Foundation of China(NSFC,Nos.52272144 and 22205048)Heilongjiang Provincial Natural Science Foundation of China(No.JQ2022E001)+3 种基金China Postdoctoral Science Foundation(Nos.2022M710931 and 2023T160154)Heilongjiang Postdoctoral Science Foundation(No.LBH-Z22010)Natural Science Foundation of Shandong Province(No.ZR2020ZD42)the Fundamental Research funds for the Central Universities are greatly acknowledged.
文摘Lateral flow immunoassay(LFIA),a rapid detection technique noted for simplicity and economy,has showcased indispensable applicability in diverse domains such as disease screening,food safety,and environmental monitoring.Nevertheless,challenges still exist in detecting ultra-low concentration analytes due to the inherent sensitivity limitations of LFIA.Recently,significant advances have been achieved by integrating enzyme activity probes and transforming LFIA into a highly sensitive tool for rapidly detecting trace analyte concentrations.Specifically,modifying natural enzymes or engineered nanozymes allows them to function as immune probes,directly catalyzing the production of signal molecules or indirectly initiating enzyme activity.Therefore,the signal intensity and detection sensitivity of LFIA are markedly elevated.The present review undertakes a comprehensive examination of pertinent research literature,offering a systematic analysis of recently proposed enzyme-based signal amplification strategies.By way of comparative assessment,the merits and demerits of current approaches are delineated,along with the identification of research avenues that still need to be explored.It is anticipated that this critical overview will garner considerable attention within the biomedical and materials science communities,providing valuable direction and insight toward the advancement of high-performance LFIA technologies.
文摘The purpose of this article is to analyze the efficacy of molecular diagnostic techniques for monitoring tuberculosis relapse.After analyzing the connotation of molecular diagnostic techniques and their specific application process in tuberculosis relapse monitoring,a total of 200 cured tuberculosis patients were selected(100 in the experimental group and 100 in the control group).During the 12-month follow-up period,the experimental group was monitored by molecular diagnostic techniques,while the control group was monitored by traditional techniques.Finally,by comparing the performance indicators of the two monitoring methods,as well as the relapse situations and outcomes of patients,it was demonstrated that molecular diagnostic techniques have higher efficacy in tuberculosis relapse monitoring.
基金supported by the National Key Research and Development Program of China(Nos.2023YFC2307305,2021YFF0703300)the Shenzhen Medical Research Fund(No.B2303003)+3 种基金Shenzhen Research Funding Program(Nos.JCYJ20220818102014028,RCBS20210609104339043)National Natural Science Foundation of China(No.22174167)Guangdong Basic and Applied Basic Research(No.2024A1515011281)Fundamental Research Funds for the Central Universities(No.24qnpy087)from Sun Yat-sen University。
文摘Ultrasensitive detection of nucleic acids is of great significance for precision medicine.Digital polymerase chain reaction(dPCR)is the most sensitive method but requires sophisticated and expensive instruments and a long reaction time.Digital PCR-free technologies,which mean the digital assay not relying on thermal cycling to amplify the signal for quantitative detection of nucleic acids at the singlemolecule level,include the digital isothermal amplification techniques(d IATs)and the digital clustered regularly interspaced short palindromic repeats(CRISPR)technologies.They combine the advantages of d PCR and IATs,which could be fast and simple,enabling absolute quantification of nucleic acids at a single-molecule level with minimum instrument,representing the next-generation molecular diagnostic technology.Herein,we systematically summarized the strategies and applications of various dIATs,including the digital loop-mediated isothermal amplification(dLAMP),the digital recombinase polymerase amplification(dRPA),the digital rolling circle amplification(dRCA),the digital nucleic acid sequencebased amplification(d NASBA)and the digital multiple displacement amplification(d MDA),and evaluated the pros and cons of each method.The emerging digital CRISPR technologies,including the detection mechanism of CRISPR and the various strategies for signal amplification,are also introduced comprehensively in this review.The current challenges as well as the future perspectives of the digital PCR-free technology were discussed.
基金Supported by Natural Science Foundation of Guangdong Province(2025A1515011061)Outstanding Graduate Entering Laboratory Project of College of Fisheries,Guangdong Ocean University+1 种基金Undergraduate Innovation and Entrepreneurship Training Program of Guangdong Ocean University(CXXL2024007)Undergraduate Innovation Team of Guangdong Ocean University(CCTD201802).
文摘[Objectives]To develop a pair of specific primers for the PCR amplification of the full-length relA gene from Vibrio alginolyticus strain HY9901,as well as to conduct bioinformatics analysis.[Methods]The relA gene was amplified through PCR,and the resulting gene sequence was subsequently analyzed using bioinformatics tools,including amino acid sequence prediction,functional site analysis,subcellular localization prediction,and homology comparison.[Results]The relA gene had a total length of 2220 bp and encoded 739 amino acid residues.The molecular weight was approximately 84.1261 kDa,and its isoelectric point was 5.95.The protein lacked a signal peptide and transmembrane regions,while exhibiting multiple phosphorylation sites.Predictions regarding its subcellular localization suggested that it was predominantly situated in the cytoplasm.The amino acid sequence demonstrated a homology of 97%to 99%with other species within the genus Vibrio,and it clustered within the same subfamily as V.antiquarius and V.diabolicus.In the prediction of secondary structure,the proportions ofα-helix,extended strand,random coil,andβ-sheet were 54.13%,12.04%,28.15%and 5.68%,respectively.The similarity between the tertiary structure model and template 5kpw.1.w was 66%.[Conclusions]In this study,the relA gene of V.alginolyticus strain HY9901 has been successfully amplified and analyzed.The structural characteristics and potential functions of the encoded protein have been elucidated,thereby providing foundational data for understanding the role of this gene in V.alginolyticus.
基金supported by National Natural Science Foundation of China(Nos.22474124,21575125)the National Natural Science Foundation of Jiangsu Province(No.BK20221370)+4 种基金Key University Natural Science Foundation of Jiangsu-Province(No.20KJA150004)the Project for Science and Technology of Yangzhou(No.YZ2022074)Project for Yangzhou City and Yangzhou University corporation(No.YZ2023204)the Open Research Fund of State Key Laboratory of Analytical Chemistry for Life Science(No.SKLACLS2405)Postgraduate Research&Practice Innovation Program of Jiangsu Province(No.KYCX22_3462)。
文摘Although diverse signal-amplified methods have been committed to improve the sensitivity of surface plasmon resonance(SPR)biosensing,introducing convenient and robust signal amplification strategy into SPR biosensing remains challenging.Here,a novel nanozyme-triggered polymerization amplification strategy was proposed for constructing highly sensitive surface plasmon resonance(SPR)immunosensor.In detail,Au@Pd core-shell nanooctahedra nanozyme with superior peroxidase(POD)-like activity was synthesized and utilized as a label probe.Simultaneously,Au@Pd core-shell nanooctahedra nanozyme can catalyze the decomposition of H_(2)O_(2)to form hydroxyl radicals(·OH)that triggers the polymerization of aniline to form polyaniline attaching on the surface of sensor chip,significantly amplifying SPR responses.The sensitivity of SPR immunosensor was enhanced by nanozyme-triggered polymerization amplification strategy.Using human immunoglobulin G(HIgG)as a model,the constructed SPR immunosensor obtains a wide linear range of 0.005–1.0μg/m L with low detection limit of 0.106 ng/m L.This research provides new sights on establishing sensitive SPR immunosensor and may evokes more inspiration for developing signal amplification methods based on nanozyme in biosensing.
基金supported by the National Natural Science Foundation of China(Grant Nos.62227821,62025503,and 62205199).
文摘To capture the nonlinear dynamics and gain evolution in chirped pulse amplification(CPA)systems,the split-step Fourier method and the fourth-order Runge–Kutta method are integrated to iteratively address the generalized nonlinear Schrödinger equation and the rate equations.However,this approach is burdened by substantial computational demands,resulting in significant time expenditures.In the context of intelligent laser optimization and inverse design,the necessity for numerous simulations further exacerbates this issue,highlighting the need for fast and accurate simulation methodologies.Here,we introduce an end-to-end model augmented with active learning(E2E-AL)with decent generalization through different dedicated embedding methods over various parameters.On an identical computational platform,the artificial intelligence–driven model is 2000 times faster than the conventional simulation method.Benefiting from the active learning strategy,the E2E-AL model achieves decent precision with only two-thirds of the training samples compared with the case without such a strategy.Furthermore,we demonstrate a multi-objective inverse design of the CPA systems enabled by the E2E-AL model.The E2E-AL framework manifests the potential of becoming a standard approach for the rapid and accurate modeling of ultrafast lasers and is readily extended to simulate other complex systems.
基金supported by the National Natural Science Foundation of China(Nos.21874060 and 22174058,U21A20282)the Science and Technology program of Gansu Province(No.22JR5RA476)。
文摘DNA repair enzymes are important in the repair of DNA lesions for maintaining the genome stability,and their abnormal expression induced various human cancers.Simultaneous detection of these DNA enzymes could provide convincing evidence based on the comparison of the activity of multiple enzymes than on that of single enzyme.Although fluorescence approach has been applied for the simultaneous detection both of DNA repair enzymes,the spectral overlap and multiwavelength excitation severely restrict the number of available fluorophores.Thus,it is difficult to simultaneously detect three enzymes in a single analysis by fluorescence detection.Herein,we developed a method for the simultaneous determination of three DNA repair enzymes including human flap DNA endonuclease 1(FEN1),human alkyladenine DNA glycosylase(hAAG)and uracil DNA glycosylase(UDG)based on the combination of template-free amplification system with capillary electrophoresis-laser induced fluorescence(CE-LIF)detection.The amplification system was adopted to transfer and amplify the enzymatic products into different length DNA fragments which could be separated effectively by CE-LIF without the complicated modification of the capillary inner wall or labeling different tails on signal probes for separation.The method demonstrated a detection limit of 0.07 U/mL(0.08-160 U/mL)for FEN1,2.40 U/mL(2.5-250U/mL)for hAAG and 2.1×10^(-4)U/mL(0.0004-2.5 U/mL)for UDG,the relative standard deviations(RSDs)of peak time and peak area for different analytes were as follows:2.50%-4,37%and 3.24%-7.18%(inter-day);1.37%-2.71%and 1.43%-3.02%(intra-day),4.28%-6.08%and 4.16%-7.57%(column to column),respectively.And it can identify the inhibitor-like drugs,evaluate enzymatic kinetics and achieve the detection of three enzymes in cell extracts,providing a simple and powerful platform for simultaneous detection of more DNA repair enzymes.
基金financially supported by the National Science and Technology Innovation 2030 Grants(2021ZD0201600)the National Key R&D Program of China(2021YFA0717000)+2 种基金the Intramural Joint Program Fund of State Key Laboratory of Microbial Technology(Project No.SKLMTIJP-2024-05)the Natural Science Foundation of Qingdao-Original exploration project(Project No.24-4-4-zrjj-139-jch)the National Natural Science Foundation of China(31771380)。
文摘Amplification-free,highly sensitive,and specific nucleic acid detection is crucial for health monitoring and diagnosis.The type III CRISPR-Cas10 system,which provides viral immunity through CRISPRassociated protein effectors,enables a new amplification-free nucleic acid diagnostic tool.In this study,we develop a CRISPR-graphene field-effect transistors(GFETs)biosensor by combining the type III CRISPR-Cas10 system with GFETs for direct nucleic acid detection.This biosensor exploits the target RNA-activated continuous ss DNA cleavage activity of the d Csm3 CRISPR-Cas10 effector and the high charge density of a hairpin DNA reporter on the GFET channel to achieve label-free,amplification-free,highly sensitive,and specific RNA detection.The CRISPR-GFET biosensor exhibits excellent performance in detecting medium-length RNAs and miRNAs,with detection limits at the aM level and a broad linear range of 10^(-15)to 10^(-11)M for RNAs and 10^(-15)to 10^(-9)M for miRNAs.It shows high sensitivity in throat swabs and serum samples,distinguishing between healthy individuals(N=5)and breast cancer patients(N=6)without the need for extraction,purification,or amplification.This platform mitigates risks associated with nucleic acid amplification and cross-contamination,making it a versatile and scalable diagnostic tool for molecular diagnostics in human health.
基金supported by the Zhejiang Provincial Natural Science Foundation of China(Grant Nos.LY23C130004 and LZ24C130004)the National Natural Science Foundation of China(Grant No.32472115)+1 种基金the National Key Research and Development Program of China(Grant No.2022YFF1003301)the Agricultural Sciences and Technologies Innovation Program of Chinese Academy of Agricultural Sciences。
文摘Bacterial blight(BB) is a devastating worldwide rice disease caused by Xanthomonas oryzae pv. oryzae(Xoo), which is difficult to diagnose based on early symptoms. Conventional chemical control yields limited effectiveness once BB has spread. Consequently, it is imperative to develop a rapid, highly sensitive, specific, and easy-to-use detection technique for early on-site diagnosis of BB. We first developed a recombinase-aided amplification-lateral flow dipstick(RAA-LFD) technique for the on-site detection of Xoo. The optimized reaction temperature and time were 37 ℃ and 20 min, indicating that the reaction system can be initiated by body temperature independently of any precision instruments. Evaluation of the RAA-LFD technique using the primers(RAAF2/R2) and probe(RAA2-nfo-probe) derived from the Xoo ORF0080 locus exhibited high specificity and eliminated cross-reactivity with other bacterial species. The sensitivity of RAA-LFD is up to 1 pg/μL for Xoo genomic DNA and 100 CFU/m L for Xoo cells. Significantly, this technique accurately detected Xoo from both artificially inoculated and naturally infected rice leaves at the early stage of infection, directly deploying plant tissue fluid as the template without DNA extraction. These attributes make the developed RAA-LFD system a viable technique for the early diagnosis of BB in the field, providing technical support for early-warning systems and disease control.
基金financial support from the National Natural Science Foundation of China(Nos.22274022 and 21874022).
文摘Development of accurate analytical protocols for cancer biomarkers is used for the initial prescreening of malignant tumors,disease surveillance,and efficacy assessment with significant clinical benefits.In this work,we reported a liposome-mediated signal-off photoelectrochemical(PEC)immunoassay for the sensitive detection of carcinoembryonic antigen(CEA)using ternary transition metal sulfide CuS/ZnCdS as the photoactive material.Good photocurrents were acquired on the basis of specific oxidation reaction of dopamine on the CuS/ZnCdS.The energy band relationship of CuS/ZnCdS was determined,and the wellmatched oxidation potential of dopamine was verified.To achieve accurate recovery of low-abundance CEA,systematic PEC evaluation from human serum samples was performed by combining with classical immunoreaction and liposome-induced dopamine amplification strategy with high stability and selectivity.Under optimum conditions,PEC immunoassay displayed good photocurrent responses toward target CEA with a dynamic linear range of 0.1-50 ng/mL with a detection limit of 31.6 pg/mL.Importantly,this system by combining with a discussion of energy level matching between semiconductor energy bands and small-molecules opens a new horizon for development of high-efficient PEC immunoassays.
