Mycobacterium marinum can cause disease in both human beings and fishes.Infected fish exhibits symptoms commonly referred to as fish tuberculosis.In this research,we screened a 14-Membered Resorcylic Acid Lactones(RAL...Mycobacterium marinum can cause disease in both human beings and fishes.Infected fish exhibits symptoms commonly referred to as fish tuberculosis.In this research,we screened a 14-Membered Resorcylic Acid Lactones(RALs)library constructed in the laboratory for in vitro anti-Mycobacterium marinum activity and found that several compounds,including compounds 3,4,6,10,and 17–26,showed good activity.Notably,the trace natural product cochilimycin B(compound 3)possesses significant long-lasting inhibitory function to Mycobacterium marinum.It also shows synergistic effects with drugs like isoniazid,ethambutol,and bedaquiline(FICI≤0.5).In addition,cochilimycin B showed a significant inhibitory effect on the biofilm produced by Mycobacterium marinum,with an inhibition rate of 65.9%.To solve the bottleneck problem of natural product drug sources,we realized an efficient one-step semi-synthetic method to obtain compound 3 with a yield of 32.1%.In vivo,inflammatory changes in BALB/c mice infected with Mycobacterium marinum were characterized.After 12 h of infection,IL-1βwas 20%–30%lower in the spleen and tail of the3-treated group compared to the control group.In the thymus and tail of the treated group,TNF-αlevels were 30%–60%lower than in the control group.This is the first time that the significant anti-Mycobacterium marinum effect of cochilimycin B from 14-Membered Resorcylic Acid Lactones(14-RALs)was reported,offering a new chemical molecule for anti-Mycobacterium marinum drug research.展开更多
Nasopharyngeal carcinoma(NPC)is a malignant tumor prevalent in southern China and Southeast Asia,where its early detection is crucial for improving patient prognosis and reducing mortality rates.However,existing scree...Nasopharyngeal carcinoma(NPC)is a malignant tumor prevalent in southern China and Southeast Asia,where its early detection is crucial for improving patient prognosis and reducing mortality rates.However,existing screening methods suffer from limitations in accuracy and accessibility,hindering their application in large-scale population screening.In this work,a surface-enhanced Raman spectroscopy(SERS)-based method was established to explore the profiles of different stratified components in saliva from NPC and healthy subjects after fractionation processing.The study findings indicate that all fractionated samples exhibit diseaseassociated molecular signaling differences,where small-molecule(molecular weight cut-offvalue is 10 kDa)demonstrating superior classification capabilities with sensitivity of 90.5%and speci-ficity of 75.6%,area under receiver operating characteristic(ROC)curve of 0:925±0:031.The primary objective of this study was to qualitatively explore patterns in saliva composition across groups.The proposed SERS detection strategy for fractionated saliva offers novel insights for enhancing the sensitivity and reliability of noninvasive NPC screening,laying the foundation for translational application in large-scale clinical settings.展开更多
Soybean(Glycine max L.)is a globally vital crop for oil production and food security.High-quality genomic resources are instrumental for both functional genomics and breeding.Here,we report a near-complete,high-qualit...Soybean(Glycine max L.)is a globally vital crop for oil production and food security.High-quality genomic resources are instrumental for both functional genomics and breeding.Here,we report a near-complete,high-quality genome assembly of the elite cultivar Tianlong 1(TL1),featuring fully resolved telomeres and centromeres,as well as a gap-free assembly of 14 of its 20 chromosomes.On the basis of the genome assembly,we generate an ethyl methanesulfonate(EMS)-mutagenized population comprising 2555 M7 plants.Whole-genome resequencing of 288 EMS mutants uncovers 1,163,869 high-confidence single-nucleotide polymorphisms(SNPs)and 542,709 insertions/deletions(InDels),achieving 91.89%coverage of predicted protein-coding genes.Phenotypic screening demonstrates robust genotype–phenotype associations,with two nonsynonymous mutants displaying pronounced defects in seed and leaf development.Collectively,the chromosome-scale TL1 genome assembly and the extensively characterized mutant population establish valuable resources for functional genomics and precision breeding in soybean and related legume species.展开更多
With the increasing accumulation of plastic pollutants in various environments,research on microorganisms(including bacteria,fungi,and algae)with plastic degradation capabilities has gained significant attention.Howev...With the increasing accumulation of plastic pollutants in various environments,research on microorganisms(including bacteria,fungi,and algae)with plastic degradation capabilities has gained significant attention.However,only a limited number of microbial plastic-degrading enzymes have been identified to date.This highlights that the degradation mechanisms employed by many plastic-degrading microorganisms,particularly filamentous fungi,remain insufficiently explored.In this study,we utilized a versatile fungal plasmid(pCT74)to express green fluorescent protein(GFP)in a marine-derived fungus Alternaria alternata strain FB1 with plastic degradation capabilities.Upon evaluating the degradation effect of polyester-type polyurethane(PU)film,we observed that different transformants exhibited three kinds of activities(the same,reduced,or enhanced degradation capability)compared to the FB1 wild-type strain.Further analysis of the plasmid fragment insertion sites in different transformants revealed that pCT74 integrates randomly into the genome of the host fungus.Notably,a direct correlation was found between the plasmid insertion site and the degradation capability of the corresponding transformant.Our findings not only redefine the potential applications of plasmid pCT74 in filamentous fungi but also show a novel research approach to identifying key enzymes involved in plastic degradation by fungi.展开更多
Insects represent emerging sources of bioactive peptides and functional materials.Mantidis Oötheca(Sang-Piao-Xiao in Chinese,SPX)serves as an insect-derived medicine for treating kidney disease.This study demonst...Insects represent emerging sources of bioactive peptides and functional materials.Mantidis Oötheca(Sang-Piao-Xiao in Chinese,SPX)serves as an insect-derived medicine for treating kidney disease.This study demonstrated that supernatant(SPX)improved kidney function in adriamycin(ADR)-induced nephropathy mice model.Transcriptomic analysis revealed that SPX inhibited complement activation by targeting the MASP1-C3/C3a receptor(C3aR)pathway.Peptidomic analysis identified 304 peptides from SPX,with 49 peptides selected for evaluation using prediction tools and molecular docking with complement core protein C3.Three peptides(PMGFPFDR,FNDPK,AAQFFNR)exhibiting docking scores below-8.0 were synthesized to verify complement inhibition and anti-fibrotic activities.The synthetic peptide AAQFFNR demonstrated complement inhibitory activity,with an inhibitory complement hemolytic 50%(ICH_(50))value of 24.54μmol·L^(-1),and exhibited superior protective effects in ADR-induced HK-2 cells.Surface plasmon resonance(SPR)assay revealed direct interaction between AAQFFNR and complement C3 with K_(d)value of 16.8μmol·L^(-1).The reno-protective effect of AAQFFNR was subsequently verified in ADR-induced mice.This research provides initial evidence that complement C3-inhibiting peptides from insects demonstrate potential in preventing nephropathy through in silico and in vivo validation approaches.展开更多
This review highlights advances in inner ear organoids(IEOs)as a novel platform for drug screening and disease modeling,particularly for hearing loss.IEOs,derived from embryonic stem cells,induced pluripotent stem cel...This review highlights advances in inner ear organoids(IEOs)as a novel platform for drug screening and disease modeling,particularly for hearing loss.IEOs,derived from embryonic stem cells,induced pluripotent stem cells,or tissue-specific progenitors,provide a physiologically relevant alternative to traditional animal models.Significant progress has been made in utilizing various cell sources,extracellular matrix materials such as Matrigel and hydrogels,and methods for controlling microenvironments through biochemical and biophysical signals.Applications of IEOs in drug screening,disease modeling,and personalized medicine enable exploration of hearing loss mechanisms and therapeutic testing.However,challenges remain,including the incomplete maturation of cochlear cells and difficulty replicating in vivo environments.Future research should focus on optimizing IEO generation,incorporating microfluidic technologies,and advancing high-throughput screening to enhance drug discovery and clinical translation.展开更多
Diabetic retinopathy(DR)is a leading cause of vision loss among working-age populations,with early screening significantly reducing the risk of blindness.However,resource-limited regions often face challenges in DR sc...Diabetic retinopathy(DR)is a leading cause of vision loss among working-age populations,with early screening significantly reducing the risk of blindness.However,resource-limited regions often face challenges in DR screening due to a shortage of ophthalmologists.This study reports the implementation and outcomes of the Chinese local standard DB52/T 1726-2023,Regulations for the application of diabetic retinopathy screening artificial intelligence,in Cambodian healthcare institutions.A pilot DR screening program with independent operational capability is established by providing a non-mydriatic fundus camera and deploying a localized diabetic retinopathy artificial intelligence(DR-AI)screening platform at the Cambodia-Kingdom Friendship Hospital in Phnom Penh,along with comprehensive training.From January to August 2025,a total of 565 patients with type 2 diabetes were screened,yielding a DR detection rate of 26.0%(147 cases).Research findings demonstrate that applying mature Chinese DR-AI screening standards and technological solutions through international collaboration in regions with a scarcity of ophthalmic professionals is both feasible and effective.This project serves as a reference for promoting DR-AI in resource-constrained countries and regions,highlighting its significant potential to leverage AI in addressing the global burden of chronic diseases and advancing the modernization of health systems.展开更多
The rational design of high-performance CO_(2)adsorbents remains a critical challenge in addressing global carbon emissions,with metal-organic frameworks(MOFs)emerging as promising candidates due to their tunable pore...The rational design of high-performance CO_(2)adsorbents remains a critical challenge in addressing global carbon emissions,with metal-organic frameworks(MOFs)emerging as promising candidates due to their tunable pore environments.However,the lack of systematic guidelines for functional group selection has hindered their practical implementation in carbon capture applications.Here,this gap was addressed by developing a comprehensive design framework through high-throughput computational screening.Through construction of a topology-directed database of 4797,integrating 10 metal centers with 144 functionalized ligands(18 ligands modified by–NH_(2),–NO_(2),–CH_(3),–CF_(3),–SH_(2),–SO_(2),–OH,and–OLi)across 36 topologies,the fundamental structure–property relationships governing CO_(2)capture performance was established.Multi-metric evaluation reveals that–NO_(2),–SO_(2),and–OLi dramatically enhance CO_(2)selectivity over CH_4/N_(2)via selectivity(S_(ads)),working capacity(ΔN),adsorbent performance score(APS),sorbent selection parameter(S_(sp)),and renewability R.Specially,ΔN rises from 2.34(pristine)to 5.91–7.94 mmol g^(-1)and S_(ads)surges from 24.94/40.36 to 121.11/176.87(–NO_(2)),149.94/215.54(–SO_(2)),and 58.64/267.44(–OLi).Besides,the critical trade-off between adsorption strength and renewability demonstrates that enhanced performance comes at the cost of reduced renewability,where stronger CO_(2)affinity(isosteric heat of-29.15,-29.96,and-30.09 for–NO_(2),–SO_(2),and–OLi)compromises renewability(R reduced by -50%).To resolve this trade-off,a novel energy efficiency(η)metric was introduced,which holistically evaluates both adsorption performance(S_(ads),ΔN,APS,S_(sp),and R)and energy inputs(desorption heat,pressure-swing energy,net loss).This leads to the identification of–SO_(2)as the optimal functional group that balances exceptional CO_(2)capture(η=6.17/12.78 for CO_(2)over CH_4/N_(2)),surpassing the second higher of 4.74/8.80 in–CF_(3)and 0.99/2.18 in non-functionalized counterparts.Adopting high-throughput computational screening methods,this work provides both fundamental insights into host–vip interactions in functionalized MOFs and a practical framework for designing next-generation adsorbents,bridging the gap between materials discovery and process engineering considerations in carbon capture technologies.展开更多
Kagome materials host intertwined phenomena,including nontrivial band topology,superconductivity,and complex charge-density-wave order,making them an important platform in condensed-matter physics and materials scienc...Kagome materials host intertwined phenomena,including nontrivial band topology,superconductivity,and complex charge-density-wave order,making them an important platform in condensed-matter physics and materials science.Motivated by extensive studies on the AV_(3)Sb_(5) family of materials,we perform high-throughput first-principles calculations to screen bilayer kagome AM_(6)X_(6) compounds with an MgFe_(6)Ge_(6)-prototype structure as potential weak-coupling superconductors.Thereafter,we systematically evaluate the thermodynamic,dynamic,and magnetic stabilities,followed by electron–phonon coupling(EPC)calculations and superconducting transition temperature estimates based on the Allen–Dynes-modified McMillan equation.From 168 candidates,we identify 31 weak-coupling superconductors that satisfy both the thermodynamic and dynamical stability criteria in our screening workflow.Focusing on compounds without partially filled f shells,we obtain superconducting transition temperatures(T_(c))of 0.65–3.97 K with EPC constants λ=0.37–0.62,indicating conventional weak-coupling superconductivity.The EPC is typically driven by vibrations within the kagome layers,with Sn-containing materials exhibiting low-frequency soft modes that contribute significantly to λ.By providing a global mapping of stability and weak-coupling superconductivity in bilayer kagome AM_(6)X_(6) compounds,this study offers a practical theoretical database and design principles for future experimental exploration.展开更多
[Objective] The study aimed to obtain attenuated strain of Haemophilus parasuis.[Method] Tn5 transposon technology was used to construct a library of mutants.Positive mutants were screened by kanamycin resistance.Fals...[Objective] The study aimed to obtain attenuated strain of Haemophilus parasuis.[Method] Tn5 transposon technology was used to construct a library of mutants.Positive mutants were screened by kanamycin resistance.False positive was identified by PCR and then removed.Mice were infected to detect the virulence of mutants.The bionomics of attenuated strains were detected,too.[Result] The attenuated mutants showed similar reproductive activity to that of wild strain.The virulence of mutants was still stable after 30 passages.[Conclusion] This study provided foundation for exploring the virulence factors and pathogenic mechanism of HPS.展开更多
The maize mutant gene Vestigial glume 1(Vg1) has been fine-mapped to a narrow region by map-based cloning and the candidate gene for Vg1 spanned 19.5 kb. Here we report Vg1 genomic fosmid library construction and scre...The maize mutant gene Vestigial glume 1(Vg1) has been fine-mapped to a narrow region by map-based cloning and the candidate gene for Vg1 spanned 19.5 kb. Here we report Vg1 genomic fosmid library construction and screening. The fosmid library of Vg1 consisted of574,000 clones with an average insert size of 36.4 kb, representing 7.9-fold coverage of the maize genome. Fosmid stability assays indicated that clones were stable during propagation in the fosmid system. Using Vg1 candidate gene-specific primers, a positive clone was successfully identified. This discovery will pave the way for identifying the function of Vg1 in maize development.展开更多
The simplicity of the CRISPR/Cas9 system of genome engineering has opened up the possibility of performing genome-wide targeted mutagenesis in cell lines,enabling screening for cellular phenotypes resulting from genet...The simplicity of the CRISPR/Cas9 system of genome engineering has opened up the possibility of performing genome-wide targeted mutagenesis in cell lines,enabling screening for cellular phenotypes resulting from genetic aberrations.Drosophila cells have proven to be highly effective in identifying genes involved in cellular processes through similar screens using partial knockdown by RNAi.This is in part due to the lower degree of redundancy between genes in this organism,whilst still maintaining highly conserved gene networks and orthologs of many human disease-causing genes.The ability of CRISPR to generate genetic loss of function mutations not only increases the magnitude of any effect over currently employed RNAi techniques,but allows analysis over longer periods of time which can be critical for certain phenotypes.In this study,we have designed and built a genome-wide CRISPR library covering 13,501 genes,among which 8989 genes are targeted by three or more independent single guide RNAs（sg RNAs）.Moreover,we describe strategies to monitor the population of guide RNAs by high throughput sequencing（HTS）.We hope that this library will provide an invaluable resource for the community to screen loss of function mutations for cellular phenotypes,and as a source of guide RNA designs for future studies.展开更多
Gayal is a rare semi-wild bovine species found in the Indo-China. They can graze grasses, including bamboo leaves, as well as reeds and other plant species, and grow to higher mature live weights than Yunnan Yellow ca...Gayal is a rare semi-wild bovine species found in the Indo-China. They can graze grasses, including bamboo leaves, as well as reeds and other plant species, and grow to higher mature live weights than Yunnan Yellow cattle maintained in similar harsh environments. The aim of this study was to identify specific cellulase in the gayal rumen. A metagenomic fosmid library was constructed using genomic DNA isolated from the ruminal contents of four adult gayals. This library contained 38400 clones with an average insert size of 35.5 kb. The Umcel-1 gene was isolated from this library. Investigation of the cellulase activity of 24 random clones led to the identification of the Umcel-1 gene, which exhibited the most potent cellulase activity. Sequencing the Umcel-1 gene revealed that it contained an open reading frame of 942 base pairs that encoded a product of 313 amino acids. The putative gene Umcel-1 product belonged to the glycosyl hydrolase family 5 and showed the highest homology to the cellulase （GenBank accession no. YP_004310852.1 ） from Clostridium lentocellum DSM 5427, with 44% identity and 62% similarity. The Umcel-1 gene was heterologously expressed in Escherichia coil BL21, and recombinant Umcel-1 was purified. The activity of purified recombinant Umcel-1 was assessed, and the results revealed that it hydrolyzed carboxymethyl cellulose with optimal activity at pH 5.5 and 45~C. To our knowledge, this study provides the first evidence for a cellulase produced by bacteria in gayal rumen.展开更多
The present study reports an improved PCR-based technique that allows quick and effective screening of cDNA libraries. First, the cDNA library was arrayed as follow: about 3 X 10’ cDNA clones were multiplied as indiv...The present study reports an improved PCR-based technique that allows quick and effective screening of cDNA libraries. First, the cDNA library was arrayed as follow: about 3 X 10’ cDNA clones were multiplied as individual plaques on solid medium in 24-well culture dishes at 1 200 plaque forming units per well. The phage suspension of each well was transferred to an individual microcentrifuge tube in 72-tube box. Then, box pool, row pools and column pools were set up that respectively represent a 72-tube box, rows and columns within the box. To screen a specific target cDNA,primers specific for novel ESTs ob- tained in our laboratory were employed to conduct PCR in a hierarchy mode. PCR began with the box pools, resulting in the identification of some Positive box pools. Then PCR went down to the row and col- umn pools of the positive box. The intersection of the positive row (s) and column (s) revealed the candi- date positive tubes. The specificity of PCR products were meanwhile checked by restriction enzyme diges- tion. Finally, hybridization was carried out to get single specific cDNA clomes from the positive tubes. This PCR-based technique features high specificity, high efficiency and less-cost in large-scale cDNA library screening. Our initial implementation of the technique resulted in the isolation of three longer different cD- NA clones from a human fetal brain cDNA library. Thus this improved technique can serve as an alterna-tive to the time-consuming and laborious conventional hybridization-based method for screening cDNA li-brary.展开更多
To study the bioactive metabolites produced by sponge-derived uncultured symbionts, a metagenomic DNA library of the symbionts of sponge Gelliodes gracilis was constructed. The average size of DNA inserts in the libra...To study the bioactive metabolites produced by sponge-derived uncultured symbionts, a metagenomic DNA library of the symbionts of sponge Gelliodes gracilis was constructed. The average size of DNA inserts in the library was 20 kb. This library was screened for antibiotic activity using paper disc assaying. Two clones displayed the antibacterial activity against Micrococcus tetragenus. The metabolites of these two clones were analyzed through HPLC. The result showed that their metabolites were quite different from those of the host E. coli DNA and the host containing vector pHZ132. This study may present a new approach to exploring bioactive metabolites of sponge symbionts.展开更多
The breeding of herbicide-resistant wheat varieties has helped control weeds in wheat fields economically and effectively.Imidazolinone (IMI) herbicides are popular as they have low toxicity in mammals,are effective a...The breeding of herbicide-resistant wheat varieties has helped control weeds in wheat fields economically and effectively.Imidazolinone (IMI) herbicides are popular as they have low toxicity in mammals,are effective at small doses,and exhibit broad-spectrum herbicidal action in the field.Therefore,the isolation and genetic and molecular characterization of IMI-resistant wheat mutants will enhance weed management in wheat fields.In the present study,352 IMI-resistant plants were isolated by genetic screening from a mutant pool prepared by EMS-based random mutagenesis.Cloning of the mutated genes from the IMI-resistant plants indicated that ten taals alleles had been isolated,and mutation in one of three Ta ALS homolog genes conferred IMI resistance,and such a mutation is a dominant trait.Further analysis showed that taals-d exhibited the greatest IMI resistance,whereas taals-b exhibited the weakest resistance to IMI among three homologous taals mutants.In terms of IMI resistance,the taals triple mutant was stronger than the taals double mutants,and the taals double mutants were stronger than the single mutants,indicating a dose-dependent effect of the Ta ALS mutation on IMI resistance in wheat.Biochemical analysis indicated that the mutation in Ta ALS increased the tolerance of Ta ALS to inhibition by IMI.Our work details the genetic and molecular characterization of als wheat mutants,provides a foundation for understanding IMI resistance and breeding wheat varieties with herbicide resistance,and indicates that genetic screening using a mutagenized pool is an effective and important means of breeding crops with additional desired agricultural traits.展开更多
The regulation of protein stability is a fundamental issue for biophysical processes,but there has not previously been a convenient and unbiased method of identifying regulators of protein stability.However,as reporte...The regulation of protein stability is a fundamental issue for biophysical processes,but there has not previously been a convenient and unbiased method of identifying regulators of protein stability.However,as reported in the article entitled "A genome-scale CRISPR-Cas9 screening method for protein stability reveals novel regulators of Cdc25 A," recently published in Cell Discovery,our team developed a protein stability regulators screening assay(Pro-SRSA) by combining the whole-genome clustered regularly interspaced short palindromic repeats Cas9(CRISPR-Cas9) library with a dual-fluorescence-based protein stability reporter and high-throughput sequencing to screen for regulators of protein stability.Based on our findings,we are confident that this efficient and unbiased screening method at the genome scale will be used by researchers worldwide to identify regulators of protein stability.展开更多
Objective:Osteosarcoma is the most common primary malignant bone tumor.However,the survival of patients with osteosarcoma has remained unchanged during the past 30 years,owing to a lack of efficient therapeutic target...Objective:Osteosarcoma is the most common primary malignant bone tumor.However,the survival of patients with osteosarcoma has remained unchanged during the past 30 years,owing to a lack of efficient therapeutic targets.Methods:We constructed a kinome-targeting CRISPR-Cas9 library containing 507 kinases and 100 nontargeting controls and screened the potential kinase targets in osteosarcoma.The CRISPR screening sequencing data were analyzed with the Model-based Analysis of Genome-wide CRISPR/Cas9 Knockout(MAGeCK)Python package.The functional data were applied in the 143B cell line through lenti-CRISPR-mediated gene knockout.The clinical significance of kinases in the survival of patients with osteosarcoma was analyzed in the R2:Genomics Analysis and Visualization Platform.Results:We identified 53 potential kinase targets in osteosarcoma.Among these targets,we analyzed 3 kinases,TRRAP,PKMYT1,and TP53RK,to validate their oncogenic functions in osteosarcoma.PKMYT1 and TP53RK showed higher expression in osteosarcoma than in normal bone tissue,whereas TRRAP showed no significant difference.High expression of all 3 kinases was associated with relatively poor prognosis in patients with osteosarcoma.Conclusions:Our results not only offer potential therapeutic kinase targets in osteosarcoma but also provide a paradigm for functional genetic screening by using a CRISPR-Cas9 library,including target design,library construction,screening workflow,data analysis,and functional validation.This method may also be useful in potentially accelerating drug discovery for other cancer types.展开更多
When devoured by macrophages,Mycobacterium tuberculosis remains persistent in macrophages and gains energy through the glyoxylate bypass to maintain its long-term existence in host cells.Therefore it is possible to st...When devoured by macrophages,Mycobacterium tuberculosis remains persistent in macrophages and gains energy through the glyoxylate bypass to maintain its long-term existence in host cells.Therefore it is possible to stop persistent infections by interdicting the glyoxylate bypass in which the isocitrate lyase（ICL） is the key rate-limiting enzyme and a persistence factor.ICL is the target of anti-TB（TB：tubercular） drugs,which could screen ICL out and effectively inhibit the activity of ICL in Mycobacterium tuberculosis,and because of this,anti-TB drugs can be used to kill persistent Mycobacterium tuberculosis.In this study,the ICL gene of the Mycobacterium tuberculosis H37Rv was cloned successfully and recombinant protein with bioactivity was obtained through the enzyme characteristic appraisal.The specific activity of the recombined ICL is 24μmol·mg-1·min-1.The recombined ICL protein was used as the target,and phages which can specifically combine to ICL were screened in the phage 7 peptide library.According to the results of the ELISA and DNA sequence detection,eventually three 7-peptide chains were synthesized.Then the peptide chains were reacted with ICL,respectively,to detect their inhibitory effects on ICL.The results show that all the three 7-peptide chains possessed varying inhibitory effects on the activity of ICL.This study provided lead compounds for the research and development of new peptide anti-TB drugs.展开更多
基金supported by the Program of National Natural Science Foundation of China(Nos.42376116,41322037,U1706210)the Special Funds of Shandong Province for Qingdao National Laboratory of Marine Science and Technology(No.2022QNLM030003-3)+1 种基金the Shandong Province Special Fund‘Frontier Technology and Free Exploration’from Laoshan Laboratory(No.8-01)the State Key Laboratory for Chemistry and Molecular Engineering of Medicinal Resources,Guangxi Normal University(No.CMEMR2023-B16)。
文摘Mycobacterium marinum can cause disease in both human beings and fishes.Infected fish exhibits symptoms commonly referred to as fish tuberculosis.In this research,we screened a 14-Membered Resorcylic Acid Lactones(RALs)library constructed in the laboratory for in vitro anti-Mycobacterium marinum activity and found that several compounds,including compounds 3,4,6,10,and 17–26,showed good activity.Notably,the trace natural product cochilimycin B(compound 3)possesses significant long-lasting inhibitory function to Mycobacterium marinum.It also shows synergistic effects with drugs like isoniazid,ethambutol,and bedaquiline(FICI≤0.5).In addition,cochilimycin B showed a significant inhibitory effect on the biofilm produced by Mycobacterium marinum,with an inhibition rate of 65.9%.To solve the bottleneck problem of natural product drug sources,we realized an efficient one-step semi-synthetic method to obtain compound 3 with a yield of 32.1%.In vivo,inflammatory changes in BALB/c mice infected with Mycobacterium marinum were characterized.After 12 h of infection,IL-1βwas 20%–30%lower in the spleen and tail of the3-treated group compared to the control group.In the thymus and tail of the treated group,TNF-αlevels were 30%–60%lower than in the control group.This is the first time that the significant anti-Mycobacterium marinum effect of cochilimycin B from 14-Membered Resorcylic Acid Lactones(14-RALs)was reported,offering a new chemical molecule for anti-Mycobacterium marinum drug research.
基金financially supported by National Natural Science Foundation ofChina(No.12374405)Provincial Science Foundation for Distinguished Young Scholars of Fujian(No.2024J010024)+1 种基金Natural Science Foundation of Fujian Province of China(No.2023J011267)Major Research Projects for Young and Middle-aged Researchers of Fujian Provincial Health Commission(No.2021ZQNZD010).
文摘Nasopharyngeal carcinoma(NPC)is a malignant tumor prevalent in southern China and Southeast Asia,where its early detection is crucial for improving patient prognosis and reducing mortality rates.However,existing screening methods suffer from limitations in accuracy and accessibility,hindering their application in large-scale population screening.In this work,a surface-enhanced Raman spectroscopy(SERS)-based method was established to explore the profiles of different stratified components in saliva from NPC and healthy subjects after fractionation processing.The study findings indicate that all fractionated samples exhibit diseaseassociated molecular signaling differences,where small-molecule(molecular weight cut-offvalue is 10 kDa)demonstrating superior classification capabilities with sensitivity of 90.5%and speci-ficity of 75.6%,area under receiver operating characteristic(ROC)curve of 0:925±0:031.The primary objective of this study was to qualitatively explore patterns in saliva composition across groups.The proposed SERS detection strategy for fractionated saliva offers novel insights for enhancing the sensitivity and reliability of noninvasive NPC screening,laying the foundation for translational application in large-scale clinical settings.
基金supported by the National Natural Science Foundation of China(31970344)Joint Funds of the Natural Science Foundation of Hainan Province,China(2021JJLH0065).
文摘Soybean(Glycine max L.)is a globally vital crop for oil production and food security.High-quality genomic resources are instrumental for both functional genomics and breeding.Here,we report a near-complete,high-quality genome assembly of the elite cultivar Tianlong 1(TL1),featuring fully resolved telomeres and centromeres,as well as a gap-free assembly of 14 of its 20 chromosomes.On the basis of the genome assembly,we generate an ethyl methanesulfonate(EMS)-mutagenized population comprising 2555 M7 plants.Whole-genome resequencing of 288 EMS mutants uncovers 1,163,869 high-confidence single-nucleotide polymorphisms(SNPs)and 542,709 insertions/deletions(InDels),achieving 91.89%coverage of predicted protein-coding genes.Phenotypic screening demonstrates robust genotype–phenotype associations,with two nonsynonymous mutants displaying pronounced defects in seed and leaf development.Collectively,the chromosome-scale TL1 genome assembly and the extensively characterized mutant population establish valuable resources for functional genomics and precision breeding in soybean and related legume species.
基金Supported by the Science and Technology Innovation Project of Laoshan Laboratory(Nos.2022QNLM030004-3,LSKJ202203103)the NSFC Innovative Group Grant(No.42221005)+5 种基金the Key Collaborative Research Program of the Alliance of International Science Organizations(No.ANSO-CR-KP-2022-08)the Shandong Provincial Natural Science Foundation(No.ZR2021ZD28)the Major Research Plan of the National Natural Science Foundation(No.92351301)the Strategic Priority Research Program of the Chinese Academy of Sciences(No.XDA22050301)the Taishan Scholars Program(No.tstp20230637)the Qingdao Natural Science Foundation(No.23-2-1-182-zyyd-jch)。
文摘With the increasing accumulation of plastic pollutants in various environments,research on microorganisms(including bacteria,fungi,and algae)with plastic degradation capabilities has gained significant attention.However,only a limited number of microbial plastic-degrading enzymes have been identified to date.This highlights that the degradation mechanisms employed by many plastic-degrading microorganisms,particularly filamentous fungi,remain insufficiently explored.In this study,we utilized a versatile fungal plasmid(pCT74)to express green fluorescent protein(GFP)in a marine-derived fungus Alternaria alternata strain FB1 with plastic degradation capabilities.Upon evaluating the degradation effect of polyester-type polyurethane(PU)film,we observed that different transformants exhibited three kinds of activities(the same,reduced,or enhanced degradation capability)compared to the FB1 wild-type strain.Further analysis of the plasmid fragment insertion sites in different transformants revealed that pCT74 integrates randomly into the genome of the host fungus.Notably,a direct correlation was found between the plasmid insertion site and the degradation capability of the corresponding transformant.Our findings not only redefine the potential applications of plasmid pCT74 in filamentous fungi but also show a novel research approach to identifying key enzymes involved in plastic degradation by fungi.
基金supported by the National Natural Science Foundation of China(No.82104353)China Postdoctoral Science Foundation funded project(No.2022M711680).
文摘Insects represent emerging sources of bioactive peptides and functional materials.Mantidis Oötheca(Sang-Piao-Xiao in Chinese,SPX)serves as an insect-derived medicine for treating kidney disease.This study demonstrated that supernatant(SPX)improved kidney function in adriamycin(ADR)-induced nephropathy mice model.Transcriptomic analysis revealed that SPX inhibited complement activation by targeting the MASP1-C3/C3a receptor(C3aR)pathway.Peptidomic analysis identified 304 peptides from SPX,with 49 peptides selected for evaluation using prediction tools and molecular docking with complement core protein C3.Three peptides(PMGFPFDR,FNDPK,AAQFFNR)exhibiting docking scores below-8.0 were synthesized to verify complement inhibition and anti-fibrotic activities.The synthetic peptide AAQFFNR demonstrated complement inhibitory activity,with an inhibitory complement hemolytic 50%(ICH_(50))value of 24.54μmol·L^(-1),and exhibited superior protective effects in ADR-induced HK-2 cells.Surface plasmon resonance(SPR)assay revealed direct interaction between AAQFFNR and complement C3 with K_(d)value of 16.8μmol·L^(-1).The reno-protective effect of AAQFFNR was subsequently verified in ADR-induced mice.This research provides initial evidence that complement C3-inhibiting peptides from insects demonstrate potential in preventing nephropathy through in silico and in vivo validation approaches.
基金supported by the National Natural Science Foundation of China(82222017 and 82271183)Hubei Province’s Key Research and Development Program(2022BCA046)the Start-up Research Fund of Southeast University(RF1028623028).
文摘This review highlights advances in inner ear organoids(IEOs)as a novel platform for drug screening and disease modeling,particularly for hearing loss.IEOs,derived from embryonic stem cells,induced pluripotent stem cells,or tissue-specific progenitors,provide a physiologically relevant alternative to traditional animal models.Significant progress has been made in utilizing various cell sources,extracellular matrix materials such as Matrigel and hydrogels,and methods for controlling microenvironments through biochemical and biophysical signals.Applications of IEOs in drug screening,disease modeling,and personalized medicine enable exploration of hearing loss mechanisms and therapeutic testing.However,challenges remain,including the incomplete maturation of cochlear cells and difficulty replicating in vivo environments.Future research should focus on optimizing IEO generation,incorporating microfluidic technologies,and advancing high-throughput screening to enhance drug discovery and clinical translation.
基金funded by the Chronic Disease Management Research Project of National Health Commission Capacity Building and Continuing Education Center 2025(GWJJMB202510024146)the Post-Subsidy Project for Standard Development of Guizhou Provincial Market Supervision and Administration Bureau 2025(DB52/T1726-2023)the Guizhou Provincial Health Commission Science and Technology Fund Project(gzwkj2024-076,gzwkj2026-146).
文摘Diabetic retinopathy(DR)is a leading cause of vision loss among working-age populations,with early screening significantly reducing the risk of blindness.However,resource-limited regions often face challenges in DR screening due to a shortage of ophthalmologists.This study reports the implementation and outcomes of the Chinese local standard DB52/T 1726-2023,Regulations for the application of diabetic retinopathy screening artificial intelligence,in Cambodian healthcare institutions.A pilot DR screening program with independent operational capability is established by providing a non-mydriatic fundus camera and deploying a localized diabetic retinopathy artificial intelligence(DR-AI)screening platform at the Cambodia-Kingdom Friendship Hospital in Phnom Penh,along with comprehensive training.From January to August 2025,a total of 565 patients with type 2 diabetes were screened,yielding a DR detection rate of 26.0%(147 cases).Research findings demonstrate that applying mature Chinese DR-AI screening standards and technological solutions through international collaboration in regions with a scarcity of ophthalmic professionals is both feasible and effective.This project serves as a reference for promoting DR-AI in resource-constrained countries and regions,highlighting its significant potential to leverage AI in addressing the global burden of chronic diseases and advancing the modernization of health systems.
基金supported by The National Natural Science Foundation of China(22471289 and 22478430)Shandong Natural Science Foundation(ZR2022ME105 and ZR2023ME004)+4 种基金Qingdao Natural Science Foundation(23-2-1-232-zyyd-jch)Geological body description and key technologies of reservoir engineering of CCUS oil displacement(2021ZZ01-03)Science and Technology Major Project on New Oil and Gas Exploration and Development:Research on Comprehensive Control Technology for CO_(2)-Enhanced Miscible and Immiscible Displacement(2024ZD1406601)State Key Laboratory of Enhanced Oil Recovery of Open Fund Funded Project(2024-KFKT-19)the Fundamental Research Funds for the Central Universities(24CX06042A and 24CX06070A)。
文摘The rational design of high-performance CO_(2)adsorbents remains a critical challenge in addressing global carbon emissions,with metal-organic frameworks(MOFs)emerging as promising candidates due to their tunable pore environments.However,the lack of systematic guidelines for functional group selection has hindered their practical implementation in carbon capture applications.Here,this gap was addressed by developing a comprehensive design framework through high-throughput computational screening.Through construction of a topology-directed database of 4797,integrating 10 metal centers with 144 functionalized ligands(18 ligands modified by–NH_(2),–NO_(2),–CH_(3),–CF_(3),–SH_(2),–SO_(2),–OH,and–OLi)across 36 topologies,the fundamental structure–property relationships governing CO_(2)capture performance was established.Multi-metric evaluation reveals that–NO_(2),–SO_(2),and–OLi dramatically enhance CO_(2)selectivity over CH_4/N_(2)via selectivity(S_(ads)),working capacity(ΔN),adsorbent performance score(APS),sorbent selection parameter(S_(sp)),and renewability R.Specially,ΔN rises from 2.34(pristine)to 5.91–7.94 mmol g^(-1)and S_(ads)surges from 24.94/40.36 to 121.11/176.87(–NO_(2)),149.94/215.54(–SO_(2)),and 58.64/267.44(–OLi).Besides,the critical trade-off between adsorption strength and renewability demonstrates that enhanced performance comes at the cost of reduced renewability,where stronger CO_(2)affinity(isosteric heat of-29.15,-29.96,and-30.09 for–NO_(2),–SO_(2),and–OLi)compromises renewability(R reduced by -50%).To resolve this trade-off,a novel energy efficiency(η)metric was introduced,which holistically evaluates both adsorption performance(S_(ads),ΔN,APS,S_(sp),and R)and energy inputs(desorption heat,pressure-swing energy,net loss).This leads to the identification of–SO_(2)as the optimal functional group that balances exceptional CO_(2)capture(η=6.17/12.78 for CO_(2)over CH_4/N_(2)),surpassing the second higher of 4.74/8.80 in–CF_(3)and 0.99/2.18 in non-functionalized counterparts.Adopting high-throughput computational screening methods,this work provides both fundamental insights into host–vip interactions in functionalized MOFs and a practical framework for designing next-generation adsorbents,bridging the gap between materials discovery and process engineering considerations in carbon capture technologies.
基金financial support from the Guangdong Provincial Quantum Science Strategic Initiative (Grant No.GDZX2501011)the Guangdong Basic and Applied Basic Research Foundation (Grant No.2024A1515010484)+7 种基金the financial support from the Guangdong Basic and Applied Basic Research Foundation (Grant No.2022A1515110404)the Guangdong Basic and Applied Basic Research Foundation (Grant No.2023A1515140188)the Guangdong Basic and Applied Basic Research Foundation (Grant No.2022A1515110322)the National Natural Science Foundation of China (Grant Nos.U2330104 and 12574028)financial support from the National Natural Science Foundation of China (Grant No.12304095)support from the National Natural Science Foundation of China (Grant No.12404190)the financial support from the National Key R&D Program of China (Grant No.2022YFA1403103)the China Postdoctoral Science Foundation (Grant No.2024M762275)。
文摘Kagome materials host intertwined phenomena,including nontrivial band topology,superconductivity,and complex charge-density-wave order,making them an important platform in condensed-matter physics and materials science.Motivated by extensive studies on the AV_(3)Sb_(5) family of materials,we perform high-throughput first-principles calculations to screen bilayer kagome AM_(6)X_(6) compounds with an MgFe_(6)Ge_(6)-prototype structure as potential weak-coupling superconductors.Thereafter,we systematically evaluate the thermodynamic,dynamic,and magnetic stabilities,followed by electron–phonon coupling(EPC)calculations and superconducting transition temperature estimates based on the Allen–Dynes-modified McMillan equation.From 168 candidates,we identify 31 weak-coupling superconductors that satisfy both the thermodynamic and dynamical stability criteria in our screening workflow.Focusing on compounds without partially filled f shells,we obtain superconducting transition temperatures(T_(c))of 0.65–3.97 K with EPC constants λ=0.37–0.62,indicating conventional weak-coupling superconductivity.The EPC is typically driven by vibrations within the kagome layers,with Sn-containing materials exhibiting low-frequency soft modes that contribute significantly to λ.By providing a global mapping of stability and weak-coupling superconductivity in bilayer kagome AM_(6)X_(6) compounds,this study offers a practical theoretical database and design principles for future experimental exploration.
基金Supported by National High Technology Research and Development Program of China(2006AA10A206)National Natural Science Foundation of China(31001072)the Fund of Beijing Academy of Agriculture and Forestry Sciences for Young Scholars(QNJJ201012)~~
文摘[Objective] The study aimed to obtain attenuated strain of Haemophilus parasuis.[Method] Tn5 transposon technology was used to construct a library of mutants.Positive mutants were screened by kanamycin resistance.False positive was identified by PCR and then removed.Mice were infected to detect the virulence of mutants.The bionomics of attenuated strains were detected,too.[Result] The attenuated mutants showed similar reproductive activity to that of wild strain.The virulence of mutants was still stable after 30 passages.[Conclusion] This study provided foundation for exploring the virulence factors and pathogenic mechanism of HPS.
基金supported by a Chongqing Postdoctoral Science Foundation funded project (Xm201344)China Postdoctoral Science Foundation funded project (2014M552303)Fundamental Research Funds for the Central Universities (XDJK2013C023, 2362015xk05)
文摘The maize mutant gene Vestigial glume 1(Vg1) has been fine-mapped to a narrow region by map-based cloning and the candidate gene for Vg1 spanned 19.5 kb. Here we report Vg1 genomic fosmid library construction and screening. The fosmid library of Vg1 consisted of574,000 clones with an average insert size of 36.4 kb, representing 7.9-fold coverage of the maize genome. Fosmid stability assays indicated that clones were stable during propagation in the fosmid system. Using Vg1 candidate gene-specific primers, a positive clone was successfully identified. This discovery will pave the way for identifying the function of Vg1 in maize development.
基金the European Research Council(DARCGENs,project number 249869)the Medical Research Council for financial support
文摘The simplicity of the CRISPR/Cas9 system of genome engineering has opened up the possibility of performing genome-wide targeted mutagenesis in cell lines,enabling screening for cellular phenotypes resulting from genetic aberrations.Drosophila cells have proven to be highly effective in identifying genes involved in cellular processes through similar screens using partial knockdown by RNAi.This is in part due to the lower degree of redundancy between genes in this organism,whilst still maintaining highly conserved gene networks and orthologs of many human disease-causing genes.The ability of CRISPR to generate genetic loss of function mutations not only increases the magnitude of any effect over currently employed RNAi techniques,but allows analysis over longer periods of time which can be critical for certain phenotypes.In this study,we have designed and built a genome-wide CRISPR library covering 13,501 genes,among which 8989 genes are targeted by three or more independent single guide RNAs（sg RNAs）.Moreover,we describe strategies to monitor the population of guide RNAs by high throughput sequencing（HTS）.We hope that this library will provide an invaluable resource for the community to screen loss of function mutations for cellular phenotypes,and as a source of guide RNA designs for future studies.
基金supported by the National Natural Science Foundation of China (31160467, 31360562, 31160449 and 31260543)
文摘Gayal is a rare semi-wild bovine species found in the Indo-China. They can graze grasses, including bamboo leaves, as well as reeds and other plant species, and grow to higher mature live weights than Yunnan Yellow cattle maintained in similar harsh environments. The aim of this study was to identify specific cellulase in the gayal rumen. A metagenomic fosmid library was constructed using genomic DNA isolated from the ruminal contents of four adult gayals. This library contained 38400 clones with an average insert size of 35.5 kb. The Umcel-1 gene was isolated from this library. Investigation of the cellulase activity of 24 random clones led to the identification of the Umcel-1 gene, which exhibited the most potent cellulase activity. Sequencing the Umcel-1 gene revealed that it contained an open reading frame of 942 base pairs that encoded a product of 313 amino acids. The putative gene Umcel-1 product belonged to the glycosyl hydrolase family 5 and showed the highest homology to the cellulase （GenBank accession no. YP_004310852.1 ） from Clostridium lentocellum DSM 5427, with 44% identity and 62% similarity. The Umcel-1 gene was heterologously expressed in Escherichia coil BL21, and recombinant Umcel-1 was purified. The activity of purified recombinant Umcel-1 was assessed, and the results revealed that it hydrolyzed carboxymethyl cellulose with optimal activity at pH 5.5 and 45~C. To our knowledge, this study provides the first evidence for a cellulase produced by bacteria in gayal rumen.
基金National Natural Science Fund!(39392900 ) 863 High-tech Project Fund of China!(102-10-03-02 )
文摘The present study reports an improved PCR-based technique that allows quick and effective screening of cDNA libraries. First, the cDNA library was arrayed as follow: about 3 X 10’ cDNA clones were multiplied as individual plaques on solid medium in 24-well culture dishes at 1 200 plaque forming units per well. The phage suspension of each well was transferred to an individual microcentrifuge tube in 72-tube box. Then, box pool, row pools and column pools were set up that respectively represent a 72-tube box, rows and columns within the box. To screen a specific target cDNA,primers specific for novel ESTs ob- tained in our laboratory were employed to conduct PCR in a hierarchy mode. PCR began with the box pools, resulting in the identification of some Positive box pools. Then PCR went down to the row and col- umn pools of the positive box. The intersection of the positive row (s) and column (s) revealed the candi- date positive tubes. The specificity of PCR products were meanwhile checked by restriction enzyme diges- tion. Finally, hybridization was carried out to get single specific cDNA clomes from the positive tubes. This PCR-based technique features high specificity, high efficiency and less-cost in large-scale cDNA library screening. Our initial implementation of the technique resulted in the isolation of three longer different cD- NA clones from a human fetal brain cDNA library. Thus this improved technique can serve as an alterna-tive to the time-consuming and laborious conventional hybridization-based method for screening cDNA li-brary.
基金This work was supported by High Tech R&D Program of China(Grant Nos.2002AA628130 and 2003AA624020)the National Natural Science Foundation of China(30171102)+2 种基金the Fund for Cheung Kong Scholar from the Cheung Kong Scholar Program of Ministry of Education of Chinathe Fund from the Natural Science Foundation of Shandong Province(No.Z2001C01)the High Tech R&D Program of Shandong Province(No.0121100107).The authors would like to thank Professor Li Jinhe of Institute of 0ceanology,Chinese Academy of Sciences,for the identification of the sponge.
文摘To study the bioactive metabolites produced by sponge-derived uncultured symbionts, a metagenomic DNA library of the symbionts of sponge Gelliodes gracilis was constructed. The average size of DNA inserts in the library was 20 kb. This library was screened for antibiotic activity using paper disc assaying. Two clones displayed the antibacterial activity against Micrococcus tetragenus. The metabolites of these two clones were analyzed through HPLC. The result showed that their metabolites were quite different from those of the host E. coli DNA and the host containing vector pHZ132. This study may present a new approach to exploring bioactive metabolites of sponge symbionts.
基金financially supported by the National Key Research and Development Program of China (2017YFD0101001)Beijing Municipal Government Science Foundation (IDHT20170513)Peking University Institute of Advanced Agricultural Sciences。
文摘The breeding of herbicide-resistant wheat varieties has helped control weeds in wheat fields economically and effectively.Imidazolinone (IMI) herbicides are popular as they have low toxicity in mammals,are effective at small doses,and exhibit broad-spectrum herbicidal action in the field.Therefore,the isolation and genetic and molecular characterization of IMI-resistant wheat mutants will enhance weed management in wheat fields.In the present study,352 IMI-resistant plants were isolated by genetic screening from a mutant pool prepared by EMS-based random mutagenesis.Cloning of the mutated genes from the IMI-resistant plants indicated that ten taals alleles had been isolated,and mutation in one of three Ta ALS homolog genes conferred IMI resistance,and such a mutation is a dominant trait.Further analysis showed that taals-d exhibited the greatest IMI resistance,whereas taals-b exhibited the weakest resistance to IMI among three homologous taals mutants.In terms of IMI resistance,the taals triple mutant was stronger than the taals double mutants,and the taals double mutants were stronger than the single mutants,indicating a dose-dependent effect of the Ta ALS mutation on IMI resistance in wheat.Biochemical analysis indicated that the mutation in Ta ALS increased the tolerance of Ta ALS to inhibition by IMI.Our work details the genetic and molecular characterization of als wheat mutants,provides a foundation for understanding IMI resistance and breeding wheat varieties with herbicide resistance,and indicates that genetic screening using a mutagenized pool is an effective and important means of breeding crops with additional desired agricultural traits.
基金supported by grants from the Key Project of Guangzhou(No.1561000151)the Yangtze River Scholarship(No.85000-52121100)+1 种基金the National Nature Science Foundation in China(No.81530081,31571395)the 973 project(No.2012CB967000)
文摘The regulation of protein stability is a fundamental issue for biophysical processes,but there has not previously been a convenient and unbiased method of identifying regulators of protein stability.However,as reported in the article entitled "A genome-scale CRISPR-Cas9 screening method for protein stability reveals novel regulators of Cdc25 A," recently published in Cell Discovery,our team developed a protein stability regulators screening assay(Pro-SRSA) by combining the whole-genome clustered regularly interspaced short palindromic repeats Cas9(CRISPR-Cas9) library with a dual-fluorescence-based protein stability reporter and high-throughput sequencing to screen for regulators of protein stability.Based on our findings,we are confident that this efficient and unbiased screening method at the genome scale will be used by researchers worldwide to identify regulators of protein stability.
基金This work was funded by the National Key Research and Development Program of China(Grant No.2016YFA0500304to T.K.)the Science and Technology Program of Guangzhou,(Grant Nos.202002020092 and 201607020038 to T.K.)+2 种基金the National Nature Science Foundation in China(NSFC)(Grant Nos.81772922 to Y.W.,81702890 to X.W.,81530081,31571395 to T.K.)the Guangdong Natural Science Foundation Team Project(Grant No.2014A030312015 to T.K.)the Natural Science Foundation of Guangdong Province(Grant No.2016A030310218 to W.Y.).
文摘Objective:Osteosarcoma is the most common primary malignant bone tumor.However,the survival of patients with osteosarcoma has remained unchanged during the past 30 years,owing to a lack of efficient therapeutic targets.Methods:We constructed a kinome-targeting CRISPR-Cas9 library containing 507 kinases and 100 nontargeting controls and screened the potential kinase targets in osteosarcoma.The CRISPR screening sequencing data were analyzed with the Model-based Analysis of Genome-wide CRISPR/Cas9 Knockout(MAGeCK)Python package.The functional data were applied in the 143B cell line through lenti-CRISPR-mediated gene knockout.The clinical significance of kinases in the survival of patients with osteosarcoma was analyzed in the R2:Genomics Analysis and Visualization Platform.Results:We identified 53 potential kinase targets in osteosarcoma.Among these targets,we analyzed 3 kinases,TRRAP,PKMYT1,and TP53RK,to validate their oncogenic functions in osteosarcoma.PKMYT1 and TP53RK showed higher expression in osteosarcoma than in normal bone tissue,whereas TRRAP showed no significant difference.High expression of all 3 kinases was associated with relatively poor prognosis in patients with osteosarcoma.Conclusions:Our results not only offer potential therapeutic kinase targets in osteosarcoma but also provide a paradigm for functional genetic screening by using a CRISPR-Cas9 library,including target design,library construction,screening workflow,data analysis,and functional validation.This method may also be useful in potentially accelerating drug discovery for other cancer types.
基金Supported by the Technology Development Funds of Education Department of Jilin Province,China(No.2008110)
文摘When devoured by macrophages,Mycobacterium tuberculosis remains persistent in macrophages and gains energy through the glyoxylate bypass to maintain its long-term existence in host cells.Therefore it is possible to stop persistent infections by interdicting the glyoxylate bypass in which the isocitrate lyase（ICL） is the key rate-limiting enzyme and a persistence factor.ICL is the target of anti-TB（TB：tubercular） drugs,which could screen ICL out and effectively inhibit the activity of ICL in Mycobacterium tuberculosis,and because of this,anti-TB drugs can be used to kill persistent Mycobacterium tuberculosis.In this study,the ICL gene of the Mycobacterium tuberculosis H37Rv was cloned successfully and recombinant protein with bioactivity was obtained through the enzyme characteristic appraisal.The specific activity of the recombined ICL is 24μmol·mg-1·min-1.The recombined ICL protein was used as the target,and phages which can specifically combine to ICL were screened in the phage 7 peptide library.According to the results of the ELISA and DNA sequence detection,eventually three 7-peptide chains were synthesized.Then the peptide chains were reacted with ICL,respectively,to detect their inhibitory effects on ICL.The results show that all the three 7-peptide chains possessed varying inhibitory effects on the activity of ICL.This study provided lead compounds for the research and development of new peptide anti-TB drugs.
