Severe fever with thrombocytopenia syndrome virus(SFTSV)is an emerging tick-borne bunyavirus that causes hemorrhagic fever-like disease(SFTS)in humans with a case fatality rate up to 30%.To date,the molecular biology ...Severe fever with thrombocytopenia syndrome virus(SFTSV)is an emerging tick-borne bunyavirus that causes hemorrhagic fever-like disease(SFTS)in humans with a case fatality rate up to 30%.To date,the molecular biology involved in SFTSV infection remains obscure.There are seven major genotypes of SFTSV(C1-C4 and J1-J3)and previously a reverse genetic system was established on a C3 strain of SFTSV.Here,we reported successfully establishment of a reverse genetics system based on a SFTSV C4 strain.First,we obtained the 5’-and 3’-terminal untranslated region(UTR)sequences of the Large(L),Medium(M)and Small(S)segments of a laboratory-adapted SFTSV C4 strain through rapid amplification of cDNA ends analysis,and developed functional T7 polymerase-based L-,M-and S-segment minigenome assays.Then,fulllength cDNA clones were constructed and infectious SFTSV were recovered from co-transfected cells.Viral infectivity,growth kinetics,and viral protein expression profile of the rescued virus were compared with the laboratory-adapted virus.Focus formation assay showed that the size and morphology of the foci formed by the rescued SFTSV were indistinguishable with the laboratory-adapted virus.However,one-step growth curve and nucleoprotein expression analyses revealed the rescued virus replicated less efficiently than the laboratory-adapted virus.Sequence analysis indicated that the difference may be due to the mutations in the laboratory-adapted strain which are more prone to cell culture.The results help us to understand the molecular biology of SFTSV,and provide a useful tool for developing vaccines and antivirals against SFTS.展开更多
To obtain the helper plasmids for a reverse genetics system of rabies virus, the cDNAs of the complete open reading frames of the N, P, G, and L genes of rabies street virus stain HN10 were each cloned into expression...To obtain the helper plasmids for a reverse genetics system of rabies virus, the cDNAs of the complete open reading frames of the N, P, G, and L genes of rabies street virus stain HN10 were each cloned into expression vector pVAX1, These four plasmids were identified by restriction enzyme digestion and gene sequencing. The plasmid encoding the N protein was selected to determine the expression effect of these plasmids in NA cells. The results showed that the helper plasmids for a reverse genetics system of rabies street virus strain HN10 had been successfully constructed.展开更多
Japanese encephalitis virus(JEV) is one of the most common pathogens of severe viral encephalitis, which is a severe threat to human health. Despite instability of the JEV genome in bacteria, many strategies have been...Japanese encephalitis virus(JEV) is one of the most common pathogens of severe viral encephalitis, which is a severe threat to human health. Despite instability of the JEV genome in bacteria, many strategies have been developed to establish molecular clone systems of JEV, providing convenient tools for studying the virus life cycle and virus–host interactions. In this study, we adapted an In-Fusion enzyme-based in vitro recombination method to construct a reverse genetic system of JEV, thereby providing a rapid approach to introduce mutations into the structural genes. A truncated genome without the structural genes was constructed as the backbone, and the complementary segment containing the structural genes was recombined in vitro, which was then transfected directly into virus-permissive cells. The progeny of the infectious virus was successfully detected in the supernatant of the transfected cells, and showed an identical phenotype to its parental virus. To provide a proof-of-principle, the 12 conserved cysteine residues in the envelope(E) protein of JEV were respectively mutated using this approach, and all mutations resulted in a complete failure to generate infectious virus. However, a leucine-tophenylanine mutation at amino acid 107 of the E protein did not interfere with the production of the infectious virus. These results suggested that all 12 cysteines in the E protein are essential for the JEV life cycle. In summary, a novel reverse genetic system of JEV was established for rapidly introducing mutations into structural genes, which will serve as a useful tool for functional studies.展开更多
The live attenuated hepatitis A virus vaccine H2 strain was developed by passaging a wild-type H2w isolate in cell cultures.Currently,the mechanism underlying its attenuation phenotype remain largely unknown.In this s...The live attenuated hepatitis A virus vaccine H2 strain was developed by passaging a wild-type H2w isolate in cell cultures.Currently,the mechanism underlying its attenuation phenotype remain largely unknown.In this study,we generated a full-length infectious cDNA clone of the H2 strain using in-fusion techniques.The recovered H2 strain(H2ic)from the cDNA clone exhibited an efficient replication in both the hepatoma cell line Huh7.5.1 and the 2BS cell line used for vaccine production,similar to the parental H2 strain.Additionally,H2ic did not cause disease in Ifnar1-/- C57 mice,consistent with the H2 strain.To explore the cell-adaptive mutations of the H2 strain,chimeric viruses were generated by replacing its non-structural proteins with corresponding regions from H2w using the infectious cDNA clone as a genetic backbone.The chimeric viruses carrying the 3C or 3D proteins from H2w showed decreased replication in Huh7.5.1 and 2BS cell lines compared to H2ic.Other chimeric viruses containing the 2B,2C,or 3A proteins from H2w failed to be recovered.Furthermore,there were no significant differences in disease manifestation in mice between H2ic and the recovered chimeric viruses.These results demonstrate that adaptive mutations in the 2B,2C,and 3A proteins are essential for efficient replication of the H2 strain in cell cultures.Mutations in the 3C and 3D proteins contribute to enhanced replication in cell cultures but did not influence the attenuated phenotypes in mice.Together,this study presents the first reverse genetic system of the H2 strain and identifies viral proteins essential for adaptation to cell cultures.展开更多
Coronavirus is an RNA virus that can infect both humans and animals,posing a significant threat to agriculture and public health.Although coronaviruses are highly host-specific,their ability to infect multiple hosts,c...Coronavirus is an RNA virus that can infect both humans and animals,posing a significant threat to agriculture and public health.Although coronaviruses are highly host-specific,their ability to infect multiple hosts,combined with the structure of their genome,gives them a high probability of genetic recombination and mutation,leading to the creation of novel viruses.In recent years,with the establishment and development of reverse genetic manipulation techniques,substantial technical support has been provided for studying the structure and function of the coronavirus genome,the development of novel vaccines and drugs and the construction of viral expression vectors.This paper briefly described the progress in research on coronaviruses and their reverse genetic system construction strategies,aiming to provide some references for future coronavirus research.展开更多
Atherosclerotic cardiovascular disease remains the leading cause of global mortality,with low-density lipoprotein cholesterol established as a primary causal risk factor.Despite widespread implementation of statin the...Atherosclerotic cardiovascular disease remains the leading cause of global mortality,with low-density lipoprotein cholesterol established as a primary causal risk factor.Despite widespread implementation of statin therapy,substantial interindividual variability in treatment response persists,necessitating precision medicine approaches to optimize therapeutic outcomes.This comprehensive narrative review synthesizes current understanding of pharmacogenomic determinants influencing lipid-lowering therapy efficacy,examines mechanisms underlying residual cardiovascular risk,and evaluates emerging therapeutic modalities targeting previously unexploited pathways in lipid metabolism.Genetic variants in key genes including 3-hydroxy-3-methylglutaryl-CoA reductase,apolipoprotein E,low-density lipoprotein receptor,and proprotein convertase subtilisin/kexin type 9 demonstrate significant associations with differential treatment responses,with specific polymorphisms conferring enhanced efficacy or increased intolerance risk.Beyond traditional statin therapy,novel therapeutic approaches targeting proprotein convertase subtilisin/kexin type 9,angiopoietin-like protein 3,apolipoprotein C-Ⅲ,and ATP citrate lyase offer substantial low-density lipoprotein cholesterol reductions of 50-80%,while RNA-based therapies including antisense oligonucleotides and small interfering RNA provide precise molecular targeting capabilities.Despite intensive lipid-lowering interventions,residual cardiovascular risk persists through four principal mechanisms:triglyceride-rich lipoproteins,lipoprotein(a),inflammatory processes,and suboptimal treatment adherence.Integration of pharmacogenomic insights with emerging therapeutic modalities enables personalized risk stratification and treatment selection,representing a paradigm shift toward precision medicine in cardiovascular disease prevention and management.展开更多
Chinese hamster with Chinese characteristics is used in experiments,and it is of great value in the field of medical biology research.However,at present,there is no high-efficiency method for evaluating the genetic qu...Chinese hamster with Chinese characteristics is used in experiments,and it is of great value in the field of medical biology research.However,at present,there is no high-efficiency method for evaluating the genetic quality of Chinese hamsters.Here,we developed a novel Chinese hamster genetic quality detection system using single-nucleotide polymorphism(SNP)markers.To find SNP loci,we conducted whole genome sequencing on 24 Chinese hamsters.Then,we employed an SNP locus screening criterion that we set up previously and initially screened 214 SNP loci with wide genome distribution and high polymorphism level.Subsequently,we developed the SNP detection system using a multitarget region capture technique based on second-generation sequencing,and a 55 SNP panel for genetic evaluation of Chinese hamster populations was developed.PopGen.32.analysis results showed that the average effective allele number,Shannon index,observed heterozygosity,expected heterozygosity,average heterozygosity,polymorphism information,and other genetic parameters of Chinese hamster population A were higher than those in population B.Using scientific screening and optimization,we successfully developed a novel Chinese hamster SNP genetic detection system that can efficiently and accurately analyze the genetic quality of the Chinese hamster population.展开更多
The polar regions host one of the harshest and most unique ecosystems on Earth.These habitats,encompassing the Arctic and the Antarctic and from deep-marine sediments to glacial ice/ice sheets,represent one of the fin...The polar regions host one of the harshest and most unique ecosystems on Earth.These habitats,encompassing the Arctic and the Antarctic and from deep-marine sediments to glacial ice/ice sheets,represent one of the final frontiers of terrestrial biological exploration.Traditionally viewed as desolate,ice-covered lands,polar regions are now recognized as vibrant,complex,and highly sensitive ecosystems.展开更多
The Polar Regions host one of the harshest and most unique ecosystems on Earth,harboring a diverse array of micro-and macro-organisms.These inhabitants showcase remarkable taxonomic and genetic originality,presenting ...The Polar Regions host one of the harshest and most unique ecosystems on Earth,harboring a diverse array of micro-and macro-organisms.These inhabitants showcase remarkable taxonomic and genetic originality,presenting unparalleled opportunities for bioprospecting,alongside demonstrating extraordinary adaptation mechanisms for survival.Furthermore,polar organisms play crucial roles in facilitating organic matter decomposition,carbon fixation and sequestration,and biogeochemical cycling.Moreover,these organisms serve as pivotal indicators of global climate shifts.Therefore,exploring the polar organisms and ecosystem holds profound and significant implications for gaining deeper insights into scientific frontiers such as global biodiversity,elementary cycling,climate change,resource utilization,and the awe of life in extreme environments.展开更多
This paper introduces an optimized planning approach for integrating photovoltaic as distributed generation (PV-DG) into the radial distribution power systems, utilizing exhaustive load flow (ELF), loss sensitivity fa...This paper introduces an optimized planning approach for integrating photovoltaic as distributed generation (PV-DG) into the radial distribution power systems, utilizing exhaustive load flow (ELF), loss sensitivity factor (LSF), genetic algorithms (GA) methods, and numerical method based on LSF. The methodology aims to determine the optimal allocation and sizing of multiple PV-DG to minimize power loss through time series power flow analysis. An approach utilizing continuous sensitivity analysis is developed and inherently leverages power flow and loss equations to compute LSF of all buses in the system towards employing a dynamic PV-DG model for more accurate results. The algorithm uses a numerical grid search method to optimize PV-DG placement in a power distribution system, focusing on minimizing system losses. It combines iterative analysis, sensitivity assessment, and comprehensive visualization to identify and present the optimal PV-DG configurations. The present-ed algorithms are verified through co-simulation framework combining MATLAB and OpenDSS to carry out analysis for 12-bus radial distribution test system. The proposed numerical method is compared with other algorithms, such as ELF, LSF methods, and Genetic Algorithms (GA). Results show that the proposed numerical method performs well in comparison with LSF and ELF solutions.展开更多
The complexity of cloud environments challenges secure resource management,especially for intrusion detection systems(IDS).Existing strategies struggle to balance efficiency,cost fairness,and threat resilience.This pa...The complexity of cloud environments challenges secure resource management,especially for intrusion detection systems(IDS).Existing strategies struggle to balance efficiency,cost fairness,and threat resilience.This paper proposes an innovative approach to managing cloud resources through the integration of a genetic algorithm(GA)with a“double auction”method.This approach seeks to enhance security and efficiency by aligning buyers and sellers within an intelligent market framework.It guarantees equitable pricing while utilizing resources efficiently and optimizing advantages for all stakeholders.The GA functions as an intelligent search mechanism that identifies optimal combinations of bids from users and suppliers,addressing issues arising from the intricacies of cloud systems.Analyses proved that our method surpasses previous strategies,particularly in terms of price accuracy,speed,and the capacity to manage large-scale activities,critical factors for real-time cybersecurity systems,such as IDS.Our research integrates artificial intelligence-inspired evolutionary algorithms with market-driven methods to develop intelligent resource management systems that are secure,scalable,and adaptable to evolving risks,such as process innovation.展开更多
The process of including renewable energy sources in power networks is moving quickly,so the need for innovative configuration solutions for grid-side ESS has grown.Among the new methods presented in this paper is GA-...The process of including renewable energy sources in power networks is moving quickly,so the need for innovative configuration solutions for grid-side ESS has grown.Among the new methods presented in this paper is GA-OCESE,which stands for Genetic Algorithm-based Optimization Configuration for Energy Storage in Electric Networks.This is one of the methods suggested in this study,which aims to enhance the sizing,positioning,and operational characteristics of structured ESS under dynamic grid conditions.Particularly,the aim is to maximize efficiency.A multiobjective genetic algorithm,the GA-OCESE framework,considers all these factors simultaneously.Besides considering cost-efficiency,response time,and energy use,the system also considers all these elements simultaneously.This enables it to effectively react to load uncertainty and variations in inputs connected to renewable sources.Results of an experimental assessment conducted on a standardized grid simulation platform indicate that by increasing energy use efficiency by 17.6%and reducing peak-load effects by 22.3%,GA-OCESE outperforms previous heuristic-based methods.This was found by contrasting the outcomes of the assessment with those of the evaluation.The results of the assessment helped to reveal this.The proposed approach will provide utility operators and energy planners with a decision-making tool that is both scalable and adaptable.This technology is particularly well-suited for smart grids,microgrid systems,and power infrastructures that heavily rely on renewable energy.Every technical component has been carefully recorded to ensure accuracy,reproducibility,and relevance across all power systems engineering software uses.This was done to ensure the program’s relevance.展开更多
Avian metapneumovirus(aMPV),a paramyxovirus,causes acute respiratory diseases in turkeys and swollen head syndrome in chickens.This study established a reverse genetics system for aMPV subtype B LN16-A strain based on...Avian metapneumovirus(aMPV),a paramyxovirus,causes acute respiratory diseases in turkeys and swollen head syndrome in chickens.This study established a reverse genetics system for aMPV subtype B LN16-A strain based on T7 RNA polymerase.Full-length cDNA of the LN16-A strain was constructed by assembling 5 cDNA fragments between the T7 promoter and hepatitis delta virus ribozyme.Transfection of this plasmid,along with the supporting plasmids encoding the N,P,M2-1,and L proteins of LN16-A into BSR-T7/5 cells,resulted in the recovery of aMPV subtype B.To identify an effective insertion site,the enhanced green fluorescent protein(EGFP)gene was inserted into different sites of the LN16-A genome to generate recombinant LN16-As.The results showed that the expression levels of EGFP at the site between the G and L genes of LN16-A were significantly higher than those at the other two sites(between the leader and N genes or replacing the SH gene).To verify the availability of the site between G and L for foreign gene expression,the VP2 gene of very virulent infectious bursal disease virus(vvIBDV)was inserted into this site,and recombinant LN16-A(rLN16A-vvVP2)was successfully rescued.Single immunization of specificpathogen-free chickens with rLN16A-vvVP2 induced high levels of neutralizing antibodies and provided 100%protection against the virulent aMPV subtype B and vvIBDV.Establishing a reverse genetics system here provides an important foundation for understanding aMPV pathogenesis and developing novel vector vaccines.展开更多
Aiming to solve the steering instability and hysteresis of agricultural robots in the process of movement,a fusion PID control method of particle swarm optimization(PSO)and genetic algorithm(GA)was proposed.The fusion...Aiming to solve the steering instability and hysteresis of agricultural robots in the process of movement,a fusion PID control method of particle swarm optimization(PSO)and genetic algorithm(GA)was proposed.The fusion algorithm took advantage of the fast optimization ability of PSO to optimize the population screening link of GA.The Simulink simulation results showed that the convergence of the fitness function of the fusion algorithm was accelerated,the system response adjustment time was reduced,and the overshoot was almost zero.Then the algorithm was applied to the steering test of agricultural robot in various scenes.After modeling the steering system of agricultural robot,the steering test results in the unloaded suspended state showed that the PID control based on fusion algorithm reduced the rise time,response adjustment time and overshoot of the system,and improved the response speed and stability of the system,compared with the artificial trial and error PID control and the PID control based on GA.The actual road steering test results showed that the PID control response rise time based on the fusion algorithm was the shortest,about 4.43 s.When the target pulse number was set to 100,the actual mean value in the steady-state regulation stage was about 102.9,which was the closest to the target value among the three control methods,and the overshoot was reduced at the same time.The steering test results under various scene states showed that the PID control based on the proposed fusion algorithm had good anti-interference ability,it can adapt to the changes of environment and load and improve the performance of the control system.It was effective in the steering control of agricultural robot.This method can provide a reference for the precise steering control of other robots.展开更多
The etiopathogenesis of gastrointestinal diseases is varied in nature.Various etiogenic factors described are infective,inflammatory,viral,bacterial,parasitic,dietary and lifestyle change.Rare causative agents are imm...The etiopathogenesis of gastrointestinal diseases is varied in nature.Various etiogenic factors described are infective,inflammatory,viral,bacterial,parasitic,dietary and lifestyle change.Rare causative agents are immunological,and others associated as idiopathic,are undiagnosed by all possible means.Some of the rare diseases are congenital in nature,passing from the parent to the child.Many of the undiagnosed diseases are now being diagnosed as genetic and the genes have been implicated as a causative agent.There is a search for newer treatments for such diseases,which is called genomic medicine.Genomic medicine is an emerging medical discipline that involves the use of genomic information about an individual.This is used both for diagnostic as well as therapeutic decisions to improve the current health domain and pave the way for policymakers for its clinical use.In the developing era of precision medicine,genomics,epigenomics,environmental exposure,and other data would be used to more accurately guide individual diagnosis and treatment.Genomic medicine is already making an impact in the fields of oncology,pharmacology,rare,infectious and many undiagnosed diseases.It is beginning to fuel new approaches in certain medical specialties.Oncology is at the leading edge of incorporating genomics,as diagnostics for genetic and genomic markers are increasingly included in cancer screening,and to guide tailored treatment strategies.Genetics and genetic medicine have been reported to play a role in gastroenterology in several ways,including genetic testing(hereditary pancreatitis and hereditary gastrointestinal cancer syndromes).Genetic testing can also help subtype diseases,such as classifying pancreatitis as idiopathic or hereditary.Gene therapy is a promising approach for treating gastrointestinal diseases that are not effectively treated by conventional pharmaceuticals and surgeries.Gene therapy strategies include gene addition,gene editing,messenger RNA therapy,and gene silencing.Understanding genetic determinants,advances in genetics,have led to a better understanding of the genetic factors that contribute to human disease.Family-member risk stratification and genetic diagnosis can help identify family members who are at risk,which can lead to preventive treatments,lifestyle recommendations,and routine follow ups.Selecting target genes helps identify the gene targets associated with each gastrointestinal disease.Common gastrointestinal diseases associated with genetic abnormalities include-inflammatory bowel disease,gastroesophageal reflux disease,non-alcoholic fatty liver disease,and irritable bowel syndrome.With advancing tools and technology,research in the search of newer and individualized treatment,genes and genetic medicines are expected to play a significant role in human health and gastroenterology.展开更多
Successful ex situ conservation of plant populations requires a high degree of genetic representativeness.However,spatially biased sampling in ex situ conservation efforts may fail to capture all wild genetic clusters...Successful ex situ conservation of plant populations requires a high degree of genetic representativeness.However,spatially biased sampling in ex situ conservation efforts may fail to capture all wild genetic clusters for species with range-wide genetic structure.To investigate the extent of spatially biased sampling in living collections and the coverage of wild genetic clusters in plant populations under ex situ conservation worldwide,we combined a global synthesis of ex situ conservation efforts with a case study of an endangered riparian plant species,Myricaria laxiflora.Our analysis of ex situ conservation worldwide revealed that the majority(82.6%)of ex situ populations fail to cover all wild genetic clusters,largely due to spatially biased sampling with low geographic coverage.Our case study of M.laxiflora showed that genetic diversity differed between the ex situ and upstream populations,while it was comparable between ex situ populations and other wild populations.However,current ex situ populations did not cover all wild genetic clusters,as the upstream genetic cluster was previously uncollected.Our study suggests that the failure to cover all wild genetic clusters in ex situ populations is a widespread issue,and ex situ populations with high genetic diversity can also fail to cover all wild genetic clusters.In future ex situ conservation programs,both the importance of high genetic diversity and the high coverage of wild genetic clusters should be prioritized.展开更多
Hereditary cardiomyopathies and arrhythmias are major contributors to cardiovascular morbidity and mortality.The advent of next-generation sequencing(NGS)has made genetic testing more accessible,which is crucial for p...Hereditary cardiomyopathies and arrhythmias are major contributors to cardiovascular morbidity and mortality.The advent of next-generation sequencing(NGS)has made genetic testing more accessible,which is crucial for precise diagnosis and targeted therapeutic strategies.The aim of this study is to explore the landscape of genetic variants,the relationship between specific variants and clinical phenotypes,and the impact on clinical decision-making in China.A total of 1536 probands(median age,37 years;1025 males[66.7%])with suspected hereditary cardiomyopathy or arrhythmia(covering 15 clinical phenotypes)are recruited from 146 hospitals across 30 provinces and cities in China.Positive results are confirmed in 390 of 1536 probands,leading to a diagnostic yield of 25.4%.Forty-two and three-tenths percent(n=169)of family members carry the same variants as positive probands.Hypertrophic cardiomyopathy(HCM)and dilated cardiomyopathy(DCM)are the predominant phenotypes,with MYBPC3 variants having the highest frequency in HCM and TTN variants in DCM.In 76.9%of the positive probands,the identified variants are helpful in clinical management,family screening,and fertility.This large-scale study provides significant insights into the genetic landscape of hereditary cardiomyopathies and arrhythmias in China.展开更多
BACKGROUND Familial adenomatous polyposis(FAP)is a disorder of autosomal dominant inheritance that is responsible for around 1%of colorectal cancer(CRC)cases.AIM To determine the mutation profile of FAP-specific to th...BACKGROUND Familial adenomatous polyposis(FAP)is a disorder of autosomal dominant inheritance that is responsible for around 1%of colorectal cancer(CRC)cases.AIM To determine the mutation profile of FAP-specific to the Hungarian population.METHODS This prospective single-center study enrolled patients with clinically suspected FAP or attenuated FAP(aFAP).Whole-exome next-generation sequencing was performed to detect variants of 50 FAP priority genes and 173 CRC predisposing genes or other CRC disease-associated genes.To identify larger deletions and insertions,a multiplex amplifiable probe hybridization technique was used.The identified genes were then classified according to the American College of Medical Genetics and Genomics guidelines.RESULTS A total of 26 index patients with clinically suspected FAP(n=21)and aFAP(n=5)were enrolled.APC gene alterations were confirmed in 92.31%of the cases(region 1B deletion,n=2;whole-gene deletion,n=4;frameshift mutation,n=2;nonsense mutation,n=5,and splice mutation,n=1),with the remaining two cases having CHEK2 and MSH3 gene alterations.According to pathogenicity,21 cases had pathogenic mutations,6 cases had likely pathogenic mutations,and 16 cases had variants of unknown significance(VUS).The most frequent of the latter were the POLE(n=5)and PIEZO1(n=4)gene variants.CONCLUSION Germline mutations in the APC gene were confirmed in more than 90%of Hungarian patients with clinically suspected FAP.Although the role of VUS genes is unclear,they are highly likely to play a role in the development of CRC.展开更多
The growing recognition of the role of genetics in the development of amyotrophic lateral sclerosis is evident.However,there has yet to be a comprehensive analysis of the clinical characteristics and genetics of famil...The growing recognition of the role of genetics in the development of amyotrophic lateral sclerosis is evident.However,there has yet to be a comprehensive analysis of the clinical characteristics and genetics of familial amyotrophic lateral sclerosis in an Asian population.This study aimed to provide an in-depth analysis of the clinical features and genetic spectrum of familial amyotrophic lateral sclerosis over 15 years in a clinic-based cohort of patients from the Chinese mainland.Enrollment of 302 amyotrophic lateral sclerosis families from 28 provinces was undertaken from January 2008 to September 2023.A group-based trajectory model for disease progression based on amyotrophic lateral sclerosis Functional Rating Scale-Revised(ALSFRS-R)scores was validated using bootstrap internal validation in patients with familial amyotrophic lateral sclerosis,as well as patients with sporadic amyotrophic lateral sclerosis(matched at a 1:4 ratio,with replacement).DNA samples from 244 index patients were screened for variants in the pathogenic genes SOD1,FUS,TDP43,and C9ORF72,of which 146 were also subjected to genome-wide next-generation sequencing.Gene-level burden analysis was used to evaluate the distribution of rare variants in the cohort.We found that rapid dynamic disease progression was associated with an older age at onset,shorter diagnostic delay,lower body mass index,bulbar onset,and≥1 affected first-degree relative.Certain attributes,such as age at onset and time from onset to diagnosis,had comparable impacts on the clinical progression trajectories of both familial amyotrophic lateral sclerosis and sporadic amyotrophic lateral sclerosis.Harboring pathogenic/likely pathogenic variants in amyotrophic lateral sclerosis-causative genes reduced the age of onset of familial amyotrophic lateral sclerosis.Among the patients with familial amyotrophic lateral sclerosis,17.8%possessed≥2 pathogenic/likely pathogenic variants.Sequencing kernel association test analysis showed that the SOD1 rare variant burden(P=1.3e-15)was associated with a significant risk of familial amyotrophic lateral sclerosis.Our findings conclusively confirmed the clinical features and genetic spectrum of familial amyotrophic lateral sclerosis over 15 years in a clinical cohort from China,contributing to a deeper understanding of genotype-phenotype relationships in familial amyotrophic lateral sclerosis.This comprehensive evaluation of specific clinical characteristics,clinical prognosis,and genetic variants of amyotrophic lateral sclerosis based on detailed clinical and genetic information may lead to the development of genotype-specific treatment approaches.展开更多
Background:Tandem gene repeats naturally occur as important genomic features and determine many traits in living organisms,like human diseases and microbial productivities of target bioproducts.Methods:Here,we develop...Background:Tandem gene repeats naturally occur as important genomic features and determine many traits in living organisms,like human diseases and microbial productivities of target bioproducts.Methods:Here,we developed a bacterial type-II toxin-antitoxin-mediated method to manipulate genomic integration of tandem gene repeats in Saccharomyces cerevisiae and further visualised the evolutionary trajectories of gene repeats.We designed a tri-vector system to introduce toxin-antitoxin-driven gene amplification modules.Results:This system delivered multi-copy gene integration in the form of tandem gene repeats spontaneously and independently from toxin-antitoxin-mediated selection.Inducing the toxin(RelE)expressing via a copper(II)-inducible CUP1 promoter successfully drove the in-situ gene amplification of the antitoxin(RelB)module,resulting in~40 copies of a green fluorescence reporter gene per copy of genome.Copy-number changes,copy-number increase and copy-number decrease,and stable maintenance were visualised using the green fluorescence protein and blue chromoprotein AeBlue as reporters.Copy-number increases happened spontaneously and independent on a selection pressure.Increased copy number was quickly enriched through toxin-antitoxin-mediated selection.Conclusion:In summary,the bacterial toxin-antitoxin systems provide a flexible mechanism to manipulate gene copy number in eukaryotic cells and can be exploited for synthetic biology and metabolic engineering applications.展开更多
基金supported by grants from the National Natural Science Foundation of China(No.31900146Open Research Fund Program of the State Key Laboratory of Virology of China(No.2020IOV003)Team project of Health Commission of Hubei Province(WJ2019C003)。
文摘Severe fever with thrombocytopenia syndrome virus(SFTSV)is an emerging tick-borne bunyavirus that causes hemorrhagic fever-like disease(SFTS)in humans with a case fatality rate up to 30%.To date,the molecular biology involved in SFTSV infection remains obscure.There are seven major genotypes of SFTSV(C1-C4 and J1-J3)and previously a reverse genetic system was established on a C3 strain of SFTSV.Here,we reported successfully establishment of a reverse genetics system based on a SFTSV C4 strain.First,we obtained the 5’-and 3’-terminal untranslated region(UTR)sequences of the Large(L),Medium(M)and Small(S)segments of a laboratory-adapted SFTSV C4 strain through rapid amplification of cDNA ends analysis,and developed functional T7 polymerase-based L-,M-and S-segment minigenome assays.Then,fulllength cDNA clones were constructed and infectious SFTSV were recovered from co-transfected cells.Viral infectivity,growth kinetics,and viral protein expression profile of the rescued virus were compared with the laboratory-adapted virus.Focus formation assay showed that the size and morphology of the foci formed by the rescued SFTSV were indistinguishable with the laboratory-adapted virus.However,one-step growth curve and nucleoprotein expression analyses revealed the rescued virus replicated less efficiently than the laboratory-adapted virus.Sequence analysis indicated that the difference may be due to the mutations in the laboratory-adapted strain which are more prone to cell culture.The results help us to understand the molecular biology of SFTSV,and provide a useful tool for developing vaccines and antivirals against SFTS.
基金National High Technology Research and Development Program of China (2006AA02Z110, 2007AA02Z402)Major Program of the National Natural Science Foundation of China (30630049)
文摘To obtain the helper plasmids for a reverse genetics system of rabies virus, the cDNAs of the complete open reading frames of the N, P, G, and L genes of rabies street virus stain HN10 were each cloned into expression vector pVAX1, These four plasmids were identified by restriction enzyme digestion and gene sequencing. The plasmid encoding the N protein was selected to determine the expression effect of these plasmids in NA cells. The results showed that the helper plasmids for a reverse genetics system of rabies street virus strain HN10 had been successfully constructed.
基金supported by grants from the National Science Foundation of China (NSFC) (No.31125003 and No.31321001)the National Basic Research Program (973 Program) of China (Grants 2010CB530103)
文摘Japanese encephalitis virus(JEV) is one of the most common pathogens of severe viral encephalitis, which is a severe threat to human health. Despite instability of the JEV genome in bacteria, many strategies have been developed to establish molecular clone systems of JEV, providing convenient tools for studying the virus life cycle and virus–host interactions. In this study, we adapted an In-Fusion enzyme-based in vitro recombination method to construct a reverse genetic system of JEV, thereby providing a rapid approach to introduce mutations into the structural genes. A truncated genome without the structural genes was constructed as the backbone, and the complementary segment containing the structural genes was recombined in vitro, which was then transfected directly into virus-permissive cells. The progeny of the infectious virus was successfully detected in the supernatant of the transfected cells, and showed an identical phenotype to its parental virus. To provide a proof-of-principle, the 12 conserved cysteine residues in the envelope(E) protein of JEV were respectively mutated using this approach, and all mutations resulted in a complete failure to generate infectious virus. However, a leucine-tophenylanine mutation at amino acid 107 of the E protein did not interfere with the production of the infectious virus. These results suggested that all 12 cysteines in the E protein are essential for the JEV life cycle. In summary, a novel reverse genetic system of JEV was established for rapidly introducing mutations into structural genes, which will serve as a useful tool for functional studies.
基金funding from the National Key Research and Development Project of China(2021YFC2300202)the National Natural Science Foundation of China(82171820)+2 种基金C.-F.Q.received support from the National Science Fund for Distinguished Young Scholar(81925025)the Innovative Research Group(81621005)from the National Natural Science Foundation Chinathe Innovation Fund for Medical Sciences(No.2019-I2M-5-049)from the Chinese Academy of Medical Sciences.
文摘The live attenuated hepatitis A virus vaccine H2 strain was developed by passaging a wild-type H2w isolate in cell cultures.Currently,the mechanism underlying its attenuation phenotype remain largely unknown.In this study,we generated a full-length infectious cDNA clone of the H2 strain using in-fusion techniques.The recovered H2 strain(H2ic)from the cDNA clone exhibited an efficient replication in both the hepatoma cell line Huh7.5.1 and the 2BS cell line used for vaccine production,similar to the parental H2 strain.Additionally,H2ic did not cause disease in Ifnar1-/- C57 mice,consistent with the H2 strain.To explore the cell-adaptive mutations of the H2 strain,chimeric viruses were generated by replacing its non-structural proteins with corresponding regions from H2w using the infectious cDNA clone as a genetic backbone.The chimeric viruses carrying the 3C or 3D proteins from H2w showed decreased replication in Huh7.5.1 and 2BS cell lines compared to H2ic.Other chimeric viruses containing the 2B,2C,or 3A proteins from H2w failed to be recovered.Furthermore,there were no significant differences in disease manifestation in mice between H2ic and the recovered chimeric viruses.These results demonstrate that adaptive mutations in the 2B,2C,and 3A proteins are essential for efficient replication of the H2 strain in cell cultures.Mutations in the 3C and 3D proteins contribute to enhanced replication in cell cultures but did not influence the attenuated phenotypes in mice.Together,this study presents the first reverse genetic system of the H2 strain and identifies viral proteins essential for adaptation to cell cultures.
基金Supported by the National Natural Science Foundation of China Joint Foundation Programme(U22A20527)。
文摘Coronavirus is an RNA virus that can infect both humans and animals,posing a significant threat to agriculture and public health.Although coronaviruses are highly host-specific,their ability to infect multiple hosts,combined with the structure of their genome,gives them a high probability of genetic recombination and mutation,leading to the creation of novel viruses.In recent years,with the establishment and development of reverse genetic manipulation techniques,substantial technical support has been provided for studying the structure and function of the coronavirus genome,the development of novel vaccines and drugs and the construction of viral expression vectors.This paper briefly described the progress in research on coronaviruses and their reverse genetic system construction strategies,aiming to provide some references for future coronavirus research.
文摘Atherosclerotic cardiovascular disease remains the leading cause of global mortality,with low-density lipoprotein cholesterol established as a primary causal risk factor.Despite widespread implementation of statin therapy,substantial interindividual variability in treatment response persists,necessitating precision medicine approaches to optimize therapeutic outcomes.This comprehensive narrative review synthesizes current understanding of pharmacogenomic determinants influencing lipid-lowering therapy efficacy,examines mechanisms underlying residual cardiovascular risk,and evaluates emerging therapeutic modalities targeting previously unexploited pathways in lipid metabolism.Genetic variants in key genes including 3-hydroxy-3-methylglutaryl-CoA reductase,apolipoprotein E,low-density lipoprotein receptor,and proprotein convertase subtilisin/kexin type 9 demonstrate significant associations with differential treatment responses,with specific polymorphisms conferring enhanced efficacy or increased intolerance risk.Beyond traditional statin therapy,novel therapeutic approaches targeting proprotein convertase subtilisin/kexin type 9,angiopoietin-like protein 3,apolipoprotein C-Ⅲ,and ATP citrate lyase offer substantial low-density lipoprotein cholesterol reductions of 50-80%,while RNA-based therapies including antisense oligonucleotides and small interfering RNA provide precise molecular targeting capabilities.Despite intensive lipid-lowering interventions,residual cardiovascular risk persists through four principal mechanisms:triglyceride-rich lipoproteins,lipoprotein(a),inflammatory processes,and suboptimal treatment adherence.Integration of pharmacogenomic insights with emerging therapeutic modalities enables personalized risk stratification and treatment selection,representing a paradigm shift toward precision medicine in cardiovascular disease prevention and management.
基金National Key Research and Development Program for Young scientists,Grant/Award Number:2021YFF0703200National Natural Foundation Joint Fund for Regional Innovation and Development,Grant/Award Number:U21A20194+1 种基金National Natural Science Foundation of China,Grant/Award Number:32170540National Key Research and Development Program,Grant/Award Number:2022YFF0711005。
文摘Chinese hamster with Chinese characteristics is used in experiments,and it is of great value in the field of medical biology research.However,at present,there is no high-efficiency method for evaluating the genetic quality of Chinese hamsters.Here,we developed a novel Chinese hamster genetic quality detection system using single-nucleotide polymorphism(SNP)markers.To find SNP loci,we conducted whole genome sequencing on 24 Chinese hamsters.Then,we employed an SNP locus screening criterion that we set up previously and initially screened 214 SNP loci with wide genome distribution and high polymorphism level.Subsequently,we developed the SNP detection system using a multitarget region capture technique based on second-generation sequencing,and a 55 SNP panel for genetic evaluation of Chinese hamster populations was developed.PopGen.32.analysis results showed that the average effective allele number,Shannon index,observed heterozygosity,expected heterozygosity,average heterozygosity,polymorphism information,and other genetic parameters of Chinese hamster population A were higher than those in population B.Using scientific screening and optimization,we successfully developed a novel Chinese hamster SNP genetic detection system that can efficiently and accurately analyze the genetic quality of the Chinese hamster population.
文摘The polar regions host one of the harshest and most unique ecosystems on Earth.These habitats,encompassing the Arctic and the Antarctic and from deep-marine sediments to glacial ice/ice sheets,represent one of the final frontiers of terrestrial biological exploration.Traditionally viewed as desolate,ice-covered lands,polar regions are now recognized as vibrant,complex,and highly sensitive ecosystems.
文摘The Polar Regions host one of the harshest and most unique ecosystems on Earth,harboring a diverse array of micro-and macro-organisms.These inhabitants showcase remarkable taxonomic and genetic originality,presenting unparalleled opportunities for bioprospecting,alongside demonstrating extraordinary adaptation mechanisms for survival.Furthermore,polar organisms play crucial roles in facilitating organic matter decomposition,carbon fixation and sequestration,and biogeochemical cycling.Moreover,these organisms serve as pivotal indicators of global climate shifts.Therefore,exploring the polar organisms and ecosystem holds profound and significant implications for gaining deeper insights into scientific frontiers such as global biodiversity,elementary cycling,climate change,resource utilization,and the awe of life in extreme environments.
文摘This paper introduces an optimized planning approach for integrating photovoltaic as distributed generation (PV-DG) into the radial distribution power systems, utilizing exhaustive load flow (ELF), loss sensitivity factor (LSF), genetic algorithms (GA) methods, and numerical method based on LSF. The methodology aims to determine the optimal allocation and sizing of multiple PV-DG to minimize power loss through time series power flow analysis. An approach utilizing continuous sensitivity analysis is developed and inherently leverages power flow and loss equations to compute LSF of all buses in the system towards employing a dynamic PV-DG model for more accurate results. The algorithm uses a numerical grid search method to optimize PV-DG placement in a power distribution system, focusing on minimizing system losses. It combines iterative analysis, sensitivity assessment, and comprehensive visualization to identify and present the optimal PV-DG configurations. The present-ed algorithms are verified through co-simulation framework combining MATLAB and OpenDSS to carry out analysis for 12-bus radial distribution test system. The proposed numerical method is compared with other algorithms, such as ELF, LSF methods, and Genetic Algorithms (GA). Results show that the proposed numerical method performs well in comparison with LSF and ELF solutions.
文摘The complexity of cloud environments challenges secure resource management,especially for intrusion detection systems(IDS).Existing strategies struggle to balance efficiency,cost fairness,and threat resilience.This paper proposes an innovative approach to managing cloud resources through the integration of a genetic algorithm(GA)with a“double auction”method.This approach seeks to enhance security and efficiency by aligning buyers and sellers within an intelligent market framework.It guarantees equitable pricing while utilizing resources efficiently and optimizing advantages for all stakeholders.The GA functions as an intelligent search mechanism that identifies optimal combinations of bids from users and suppliers,addressing issues arising from the intricacies of cloud systems.Analyses proved that our method surpasses previous strategies,particularly in terms of price accuracy,speed,and the capacity to manage large-scale activities,critical factors for real-time cybersecurity systems,such as IDS.Our research integrates artificial intelligence-inspired evolutionary algorithms with market-driven methods to develop intelligent resource management systems that are secure,scalable,and adaptable to evolving risks,such as process innovation.
文摘The process of including renewable energy sources in power networks is moving quickly,so the need for innovative configuration solutions for grid-side ESS has grown.Among the new methods presented in this paper is GA-OCESE,which stands for Genetic Algorithm-based Optimization Configuration for Energy Storage in Electric Networks.This is one of the methods suggested in this study,which aims to enhance the sizing,positioning,and operational characteristics of structured ESS under dynamic grid conditions.Particularly,the aim is to maximize efficiency.A multiobjective genetic algorithm,the GA-OCESE framework,considers all these factors simultaneously.Besides considering cost-efficiency,response time,and energy use,the system also considers all these elements simultaneously.This enables it to effectively react to load uncertainty and variations in inputs connected to renewable sources.Results of an experimental assessment conducted on a standardized grid simulation platform indicate that by increasing energy use efficiency by 17.6%and reducing peak-load effects by 22.3%,GA-OCESE outperforms previous heuristic-based methods.This was found by contrasting the outcomes of the assessment with those of the evaluation.The results of the assessment helped to reveal this.The proposed approach will provide utility operators and energy planners with a decision-making tool that is both scalable and adaptable.This technology is particularly well-suited for smart grids,microgrid systems,and power infrastructures that heavily rely on renewable energy.Every technical component has been carefully recorded to ensure accuracy,reproducibility,and relevance across all power systems engineering software uses.This was done to ensure the program’s relevance.
基金supported by the grants from the National Key Research and Development Program of China(2022YFD1800604)the China Agriculture Research System(CARS-41)the Heilongjiang Touyan Innovation Team Program,China。
文摘Avian metapneumovirus(aMPV),a paramyxovirus,causes acute respiratory diseases in turkeys and swollen head syndrome in chickens.This study established a reverse genetics system for aMPV subtype B LN16-A strain based on T7 RNA polymerase.Full-length cDNA of the LN16-A strain was constructed by assembling 5 cDNA fragments between the T7 promoter and hepatitis delta virus ribozyme.Transfection of this plasmid,along with the supporting plasmids encoding the N,P,M2-1,and L proteins of LN16-A into BSR-T7/5 cells,resulted in the recovery of aMPV subtype B.To identify an effective insertion site,the enhanced green fluorescent protein(EGFP)gene was inserted into different sites of the LN16-A genome to generate recombinant LN16-As.The results showed that the expression levels of EGFP at the site between the G and L genes of LN16-A were significantly higher than those at the other two sites(between the leader and N genes or replacing the SH gene).To verify the availability of the site between G and L for foreign gene expression,the VP2 gene of very virulent infectious bursal disease virus(vvIBDV)was inserted into this site,and recombinant LN16-A(rLN16A-vvVP2)was successfully rescued.Single immunization of specificpathogen-free chickens with rLN16A-vvVP2 induced high levels of neutralizing antibodies and provided 100%protection against the virulent aMPV subtype B and vvIBDV.Establishing a reverse genetics system here provides an important foundation for understanding aMPV pathogenesis and developing novel vector vaccines.
文摘Aiming to solve the steering instability and hysteresis of agricultural robots in the process of movement,a fusion PID control method of particle swarm optimization(PSO)and genetic algorithm(GA)was proposed.The fusion algorithm took advantage of the fast optimization ability of PSO to optimize the population screening link of GA.The Simulink simulation results showed that the convergence of the fitness function of the fusion algorithm was accelerated,the system response adjustment time was reduced,and the overshoot was almost zero.Then the algorithm was applied to the steering test of agricultural robot in various scenes.After modeling the steering system of agricultural robot,the steering test results in the unloaded suspended state showed that the PID control based on fusion algorithm reduced the rise time,response adjustment time and overshoot of the system,and improved the response speed and stability of the system,compared with the artificial trial and error PID control and the PID control based on GA.The actual road steering test results showed that the PID control response rise time based on the fusion algorithm was the shortest,about 4.43 s.When the target pulse number was set to 100,the actual mean value in the steady-state regulation stage was about 102.9,which was the closest to the target value among the three control methods,and the overshoot was reduced at the same time.The steering test results under various scene states showed that the PID control based on the proposed fusion algorithm had good anti-interference ability,it can adapt to the changes of environment and load and improve the performance of the control system.It was effective in the steering control of agricultural robot.This method can provide a reference for the precise steering control of other robots.
文摘The etiopathogenesis of gastrointestinal diseases is varied in nature.Various etiogenic factors described are infective,inflammatory,viral,bacterial,parasitic,dietary and lifestyle change.Rare causative agents are immunological,and others associated as idiopathic,are undiagnosed by all possible means.Some of the rare diseases are congenital in nature,passing from the parent to the child.Many of the undiagnosed diseases are now being diagnosed as genetic and the genes have been implicated as a causative agent.There is a search for newer treatments for such diseases,which is called genomic medicine.Genomic medicine is an emerging medical discipline that involves the use of genomic information about an individual.This is used both for diagnostic as well as therapeutic decisions to improve the current health domain and pave the way for policymakers for its clinical use.In the developing era of precision medicine,genomics,epigenomics,environmental exposure,and other data would be used to more accurately guide individual diagnosis and treatment.Genomic medicine is already making an impact in the fields of oncology,pharmacology,rare,infectious and many undiagnosed diseases.It is beginning to fuel new approaches in certain medical specialties.Oncology is at the leading edge of incorporating genomics,as diagnostics for genetic and genomic markers are increasingly included in cancer screening,and to guide tailored treatment strategies.Genetics and genetic medicine have been reported to play a role in gastroenterology in several ways,including genetic testing(hereditary pancreatitis and hereditary gastrointestinal cancer syndromes).Genetic testing can also help subtype diseases,such as classifying pancreatitis as idiopathic or hereditary.Gene therapy is a promising approach for treating gastrointestinal diseases that are not effectively treated by conventional pharmaceuticals and surgeries.Gene therapy strategies include gene addition,gene editing,messenger RNA therapy,and gene silencing.Understanding genetic determinants,advances in genetics,have led to a better understanding of the genetic factors that contribute to human disease.Family-member risk stratification and genetic diagnosis can help identify family members who are at risk,which can lead to preventive treatments,lifestyle recommendations,and routine follow ups.Selecting target genes helps identify the gene targets associated with each gastrointestinal disease.Common gastrointestinal diseases associated with genetic abnormalities include-inflammatory bowel disease,gastroesophageal reflux disease,non-alcoholic fatty liver disease,and irritable bowel syndrome.With advancing tools and technology,research in the search of newer and individualized treatment,genes and genetic medicines are expected to play a significant role in human health and gastroenterology.
基金supported by National Key Research and Development Program of China(2024YFF1307400)Hubei Provincial Natural Science Foundation and Three Gorges Innovation Development Joint Fund(Grant No.2023AFD195)China Three Gorges Corporation(NBZZ202300130).
文摘Successful ex situ conservation of plant populations requires a high degree of genetic representativeness.However,spatially biased sampling in ex situ conservation efforts may fail to capture all wild genetic clusters for species with range-wide genetic structure.To investigate the extent of spatially biased sampling in living collections and the coverage of wild genetic clusters in plant populations under ex situ conservation worldwide,we combined a global synthesis of ex situ conservation efforts with a case study of an endangered riparian plant species,Myricaria laxiflora.Our analysis of ex situ conservation worldwide revealed that the majority(82.6%)of ex situ populations fail to cover all wild genetic clusters,largely due to spatially biased sampling with low geographic coverage.Our case study of M.laxiflora showed that genetic diversity differed between the ex situ and upstream populations,while it was comparable between ex situ populations and other wild populations.However,current ex situ populations did not cover all wild genetic clusters,as the upstream genetic cluster was previously uncollected.Our study suggests that the failure to cover all wild genetic clusters in ex situ populations is a widespread issue,and ex situ populations with high genetic diversity can also fail to cover all wild genetic clusters.In future ex situ conservation programs,both the importance of high genetic diversity and the high coverage of wild genetic clusters should be prioritized.
基金supported by Science,Technology&Innovation Project of Xiongan New Area(2023XAGG0069)National Key Research and Development Program of China(2022YFC2703100)+2 种基金National High Level Hospital Clinical Research Funding(2022-PUMCH-D-002)the National Natural Science Foundation of China(824B2011 to Z.W.)National High Level Hospital Clinical Research Funding(2023-PUMCH-E-012).
文摘Hereditary cardiomyopathies and arrhythmias are major contributors to cardiovascular morbidity and mortality.The advent of next-generation sequencing(NGS)has made genetic testing more accessible,which is crucial for precise diagnosis and targeted therapeutic strategies.The aim of this study is to explore the landscape of genetic variants,the relationship between specific variants and clinical phenotypes,and the impact on clinical decision-making in China.A total of 1536 probands(median age,37 years;1025 males[66.7%])with suspected hereditary cardiomyopathy or arrhythmia(covering 15 clinical phenotypes)are recruited from 146 hospitals across 30 provinces and cities in China.Positive results are confirmed in 390 of 1536 probands,leading to a diagnostic yield of 25.4%.Forty-two and three-tenths percent(n=169)of family members carry the same variants as positive probands.Hypertrophic cardiomyopathy(HCM)and dilated cardiomyopathy(DCM)are the predominant phenotypes,with MYBPC3 variants having the highest frequency in HCM and TTN variants in DCM.In 76.9%of the positive probands,the identified variants are helpful in clinical management,family screening,and fertility.This large-scale study provides significant insights into the genetic landscape of hereditary cardiomyopathies and arrhythmias in China.
基金Supported by the Research Grants of the National Research,Development and Innovation Office,No.K125377,No.K134863 and No.K143549New National Excellence Program of the Ministry of Human Capacities,No.UNKP-20-5-SZTE-161,No.UNKP-22-3-SZTE-233,No.UNKP-23-5-SZTE-719,No.UNKP-22-4-SZTE-296 and No.UNKP-22-3-SZTE-278+1 种基金Janos Bolyai Research Grant,No.BO/00723/22the Géza Hetényi Research Grant by Albert Szent-Györgyi Medical School,University of Szeged.
文摘BACKGROUND Familial adenomatous polyposis(FAP)is a disorder of autosomal dominant inheritance that is responsible for around 1%of colorectal cancer(CRC)cases.AIM To determine the mutation profile of FAP-specific to the Hungarian population.METHODS This prospective single-center study enrolled patients with clinically suspected FAP or attenuated FAP(aFAP).Whole-exome next-generation sequencing was performed to detect variants of 50 FAP priority genes and 173 CRC predisposing genes or other CRC disease-associated genes.To identify larger deletions and insertions,a multiplex amplifiable probe hybridization technique was used.The identified genes were then classified according to the American College of Medical Genetics and Genomics guidelines.RESULTS A total of 26 index patients with clinically suspected FAP(n=21)and aFAP(n=5)were enrolled.APC gene alterations were confirmed in 92.31%of the cases(region 1B deletion,n=2;whole-gene deletion,n=4;frameshift mutation,n=2;nonsense mutation,n=5,and splice mutation,n=1),with the remaining two cases having CHEK2 and MSH3 gene alterations.According to pathogenicity,21 cases had pathogenic mutations,6 cases had likely pathogenic mutations,and 16 cases had variants of unknown significance(VUS).The most frequent of the latter were the POLE(n=5)and PIEZO1(n=4)gene variants.CONCLUSION Germline mutations in the APC gene were confirmed in more than 90%of Hungarian patients with clinically suspected FAP.Although the role of VUS genes is unclear,they are highly likely to play a role in the development of CRC.
基金supported by the Natural Science Foundation of Beijing,Nos.7244428(to WZ)and 7222215(to JH)the Peking University Medicine Sailing Program forYoung Scholars’Scientific and Technological Innovation,No.BMU2023YFJHPY034(to WZ)+4 种基金the National Natural Science Foundation of China,Nos.81873784,82071426(to DF),and81974197(to JH)the Clinical Cohort Construction Program of Peking University Third Hospital,No.BYSYDL2019002(to DF)Beijing Physician-Scientist TrainingProgram,No.BJPSTP-2024-03(to JH)the China Postdoctoral Science Foundation,Nos.2022TQ0014(to LX),2022M720284(to LX)the E-Town Cooperation&Development Foundation,No.YCXJ-JZ-2023-017(to LX).
文摘The growing recognition of the role of genetics in the development of amyotrophic lateral sclerosis is evident.However,there has yet to be a comprehensive analysis of the clinical characteristics and genetics of familial amyotrophic lateral sclerosis in an Asian population.This study aimed to provide an in-depth analysis of the clinical features and genetic spectrum of familial amyotrophic lateral sclerosis over 15 years in a clinic-based cohort of patients from the Chinese mainland.Enrollment of 302 amyotrophic lateral sclerosis families from 28 provinces was undertaken from January 2008 to September 2023.A group-based trajectory model for disease progression based on amyotrophic lateral sclerosis Functional Rating Scale-Revised(ALSFRS-R)scores was validated using bootstrap internal validation in patients with familial amyotrophic lateral sclerosis,as well as patients with sporadic amyotrophic lateral sclerosis(matched at a 1:4 ratio,with replacement).DNA samples from 244 index patients were screened for variants in the pathogenic genes SOD1,FUS,TDP43,and C9ORF72,of which 146 were also subjected to genome-wide next-generation sequencing.Gene-level burden analysis was used to evaluate the distribution of rare variants in the cohort.We found that rapid dynamic disease progression was associated with an older age at onset,shorter diagnostic delay,lower body mass index,bulbar onset,and≥1 affected first-degree relative.Certain attributes,such as age at onset and time from onset to diagnosis,had comparable impacts on the clinical progression trajectories of both familial amyotrophic lateral sclerosis and sporadic amyotrophic lateral sclerosis.Harboring pathogenic/likely pathogenic variants in amyotrophic lateral sclerosis-causative genes reduced the age of onset of familial amyotrophic lateral sclerosis.Among the patients with familial amyotrophic lateral sclerosis,17.8%possessed≥2 pathogenic/likely pathogenic variants.Sequencing kernel association test analysis showed that the SOD1 rare variant burden(P=1.3e-15)was associated with a significant risk of familial amyotrophic lateral sclerosis.Our findings conclusively confirmed the clinical features and genetic spectrum of familial amyotrophic lateral sclerosis over 15 years in a clinical cohort from China,contributing to a deeper understanding of genotype-phenotype relationships in familial amyotrophic lateral sclerosis.This comprehensive evaluation of specific clinical characteristics,clinical prognosis,and genetic variants of amyotrophic lateral sclerosis based on detailed clinical and genetic information may lead to the development of genotype-specific treatment approaches.
基金supported partially by the Australian Government through the Australian Research Council Centres of Excellence funding scheme(project CE200100029)。
文摘Background:Tandem gene repeats naturally occur as important genomic features and determine many traits in living organisms,like human diseases and microbial productivities of target bioproducts.Methods:Here,we developed a bacterial type-II toxin-antitoxin-mediated method to manipulate genomic integration of tandem gene repeats in Saccharomyces cerevisiae and further visualised the evolutionary trajectories of gene repeats.We designed a tri-vector system to introduce toxin-antitoxin-driven gene amplification modules.Results:This system delivered multi-copy gene integration in the form of tandem gene repeats spontaneously and independently from toxin-antitoxin-mediated selection.Inducing the toxin(RelE)expressing via a copper(II)-inducible CUP1 promoter successfully drove the in-situ gene amplification of the antitoxin(RelB)module,resulting in~40 copies of a green fluorescence reporter gene per copy of genome.Copy-number changes,copy-number increase and copy-number decrease,and stable maintenance were visualised using the green fluorescence protein and blue chromoprotein AeBlue as reporters.Copy-number increases happened spontaneously and independent on a selection pressure.Increased copy number was quickly enriched through toxin-antitoxin-mediated selection.Conclusion:In summary,the bacterial toxin-antitoxin systems provide a flexible mechanism to manipulate gene copy number in eukaryotic cells and can be exploited for synthetic biology and metabolic engineering applications.
