Objective:Osteoarthritis(OA)and sarcopenia are significant health concerns in the elderly,substantially impacting their daily activities and quality of life.However,the relationship between them remains poorly underst...Objective:Osteoarthritis(OA)and sarcopenia are significant health concerns in the elderly,substantially impacting their daily activities and quality of life.However,the relationship between them remains poorly understood.This study aims to uncover common biomarkers and pathways associated with both OA and sarcopenia.Methods:Gene expression profiles related to OA and sarcopenia were retrieved from the Gene Expression Omnibus(GEO)database.Differentially expressed genes(DEGs)between disease and control groups were identified using R software.Common DEGs were extracted via Venn diagram analysis.Gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses were conducted to identify biological processes and pathways associated with shared DEGs.Protein-protein interaction(PPI)networks were constructed,and candidate hub genes were ranked using the maximal clique centrality(MCC)algorithm.Further validation of hub gene expression was performed using 2 independent datasets.Receiver operating characteristic(ROC)curve analysis was used to evaluate the predictive value of key genes for OA and sarcopenia.Mouse models of OA and sarcopenia were established.Hematoxylin-eosin and Safranin O/Fast Green staining were used to validate the OA model.The sarcopenia model was validated via rotarod testing and quadriceps muscle mass measurement.Real-time reverse transcription PCR(real-time RT-PCR)was employed to assess the mRNA expression levels of candidate key genes in both models.Gene set enrichment analysis(GSEA)was conducted to identify pathways associated with the selected shared key genes in both diseases.Results:A total of 89 common DEGs were identified in the gene expression profiles of OA and sarcopenia,including 76 upregulated and 13 downregulated genes.These 89 DEGs were significantly enriched in protein digestion and absorption,the PI3K-Akt signaling pathway,and extracellular matrix-receptor interaction.PPI network analysis and MCC algorithm analysis of the 89 common DEGs identified the top 17 candidate hub genes.Based on the differential expression analysis of these 17 candidate hub genes in the validation datasets,AEBP1 and COL8A2 were ultimately selected as the common key genes for both diseases,both of which showed a significant upregulation trend in the disease groups(all P<0.05).The value of area under the curve(AUC)for AEBP1 and COL8A2 in the OA and sarcopenia datasets were all greater than 0.7,indicating that both genes have potential value in predicting OA and sarcopenia.Real-time RT-PCR results showed that the mRNA expression levels of AEBP1 and COL8A2 were significantly upregulated in the disease groups(all P<0.05),consistent with the results observed in the bioinformatics analysis.GSEA revealed that AEBP1 and COL8A2 were closely related to extracellular matrix-receptor interaction,ribosome,and oxidative phosphorylation in OA and sarcopenia.Conclusion:AEBP1 and COL8A2 have the potential to serve as common biomarkers for OA and sarcopenia.The extracellular matrix-receptor interaction pathway may represent a potential target for the prevention and treatment of both OA and sarcopenia.展开更多
Objective:Keratoconus(KC)is a progressive corneal ectasia disorder,arising from a myriad of causes including genetic predispositions,environmental factors,biomechanical influences,and inflammatory reactions.This study...Objective:Keratoconus(KC)is a progressive corneal ectasia disorder,arising from a myriad of causes including genetic predispositions,environmental factors,biomechanical influences,and inflammatory reactions.This study aims to identify potential pathogenetic gene mutations in patients with sporadic KC in the Han Chinese population.Methods:Twenty-five patients with primary KC as well as 50 unrelated population matched healthy controls,were included in this study to identify potential pathogenic gene mutations among sporadic KC patients in the Han Chinese population.Sanger sequencing and whole-exome sequencing(WES)were used to analyze mutations in the zinc finger protein 469(ZNF469)gene.Bioinformatics analysis was conducted to explore the potential role of ZNF469 in KC pathogenesis.Results:Five novel heterozygous missense variants were identified in KC patients.Among them,2 compound heterozygous variants,c.8986G>C(p.E2996Q)with c.11765A>C(p.D3922A),and c.4423C>G(p.L1475V)with c.10633G>A(p.G3545R),were determined to be possible pathogenic factors for KC.Conclusion:Mutations in the ZNF469 gene may contribute to the development of KC in the Han Chinese population.These mutation sites may provide valuable information for future genetic screening of KC patients and their families.展开更多
Objective To investigate the structural changes of rat thoracic aorta and changes in expression levels of Bmal1 and cyclins in thoracic aorta endothelial cells following heat stress.Methods Twenty male SD rats were ra...Objective To investigate the structural changes of rat thoracic aorta and changes in expression levels of Bmal1 and cyclins in thoracic aorta endothelial cells following heat stress.Methods Twenty male SD rats were randomized equally into control group and heat stress group.After exposure to 32℃for 2 weeks in the latter group,the rats were examined for histopathological changes and Bmal1 expression in the thoracic aorta using HE staining and immunohistochemistry.In the cell experiments,cultured rat thoracic aortic endothelial cells(RTAECs)were incubated at 40℃for 12 h with or without prior transfection with a Bmal1-specific small interfering RNA(si-Bmal1)or a negative sequence.In both rat thoracic aorta and RTAECs,the expressions of Bmal1,the cell cycle proteins CDK1,CDK4,CDK6,and cyclin B1,and apoptosis-related proteins Bax and Bcl-2 were detected using Western blotting.TUNEL staining was used to detect cell apoptosis in rat thoracic aorta,and the changes in cell cycle distribution and apoptosis in RTAECs were analyzed with flow cytometry.Results Compared with the control rats,the rats exposed to heat stress showed significantly increased blood pressures and lowered heart rate with elastic fiber disruption and increased expressions of Bmal1,cyclin B1 and CDK1 in the thoracic aorta(P<0.05).In cultured RTAECs,heat stress caused significant increase of Bmal1,cyclin B1 and CDK1 protein expression levels,which were obviously lowered in cells with prior si-Bmal1 transfection.Bmal1 knockdown also inhibited heat stress-induced increase of apoptosis in RTAECs as evidenced by decreased expression of Bax and increased expression of Bcl-2.Conclusion Heat stress upregulates Bmal1 expression and causes alterations in expressions of cyclins to trigger apoptosis of rat thoracic aorta endothelial cells,which can be partly alleviated by suppressing Bmal1 expression.展开更多
Background Verticillium dahliae,a soil-borne fungi,can cause Verticillium wilt,and seriously diminish the yield and quality of cotton.However,the pathogenic mechanism of V.dahliae is complex and not clearly understood...Background Verticillium dahliae,a soil-borne fungi,can cause Verticillium wilt,and seriously diminish the yield and quality of cotton.However,the pathogenic mechanism of V.dahliae is complex and not clearly understood at the moment.This study aimed to identify the high-affinity nicotinic acid transporter genes in V.dahliae.The gene expression profiles in V.dahliae following sensing of root exudates from susceptible and resistant cotton varieties were analyzed.The function of VdNAT1 in the pathogenic process of V.dahliae was studied using the tobacco rattle virus(TRV)-based host-induced gene silencing(HIGS)technique.Results Eight high-affinity nicotinic acid transporter genes were identified from V.dahliae through the bioinformatics method.Each protein contains a conserved major facilitator superfamily(MFS)domain,which belongs to the MFS superfamily.Evolutionary relationship analysis revealed that all 8 genes belong to the anion:cation symporter(ACS)subfamily.All proteins have transmembrane domains,ranging from 7 to 12.The expression levels of most VdNAT genes were significantly increased after induction by root exudates from susceptible cotton varieties.Silencing VdNAT1 gene by HIGS significantly inhibited the accumulation of fungal biomass in cotton plants,and alleviated the disease symptoms of cotton.Conclusions Eight VdNAT genes were identified from V.dahliae,and most VdNAT genes was up-regulated after induced by root exudates from susceptible cotton variety.In addition,VdNAT1 is required for the pathogenicity of V.dahliae.Overall,these findings will facilitate the pathogenic molecular mechanism of V.dahliae and provide candidate genes.展开更多
The advent of gene editing represents one of the most transformative breakthroughs in life science,making genome manipulation more accessible than ever before.While traditional CRISPR/Cas-based gene editing,which invo...The advent of gene editing represents one of the most transformative breakthroughs in life science,making genome manipulation more accessible than ever before.While traditional CRISPR/Cas-based gene editing,which involves double-strand DNA breaks(DSBs),excels at gene disruption,it is less effective for accurate gene modification.The limitation arises because DSBs are primarily repaired via non-homologous end joining(NHEJ),which tends to introduce indels at the break site.While homology directed repair(HDR)can achieve precise editing when a donor DNA template is provided,the reliance on DSBs often results in unintended genome damage.HDR is restricted to specific cell cycle phases,limiting its application.Currently,gene editing has evolved to unprecedented levels of precision without relying on DSB and HDR.The development of innovative systems,such as base editing,prime editing,and CRISPR-associated transposases(CASTs),now allow for precise editing ranging from single nucleotides to large DNA fragments.Base editors(BEs)enable the direct conversion of one nucleotide to another,and prime editors(PEs)further expand gene editing capabilities by allowing for the insertion,deletion,or alteration of small DNA fragments.The CAST system,a recent innovation,allows for the precise insertion of large DNA fragments at specific genomic locations.In recent years,the optimization of these precise gene editing tools has led to significant improvements in editing efficiency,specificity,and versatility,with advancements such as the creation of base editors for nucleotide transversions,enhanced prime editing systems for more efficient and precise modifications,and refined CAST systems for targeted large DNA insertions,expanding the range of applications for these tools.Concurrently,these advances are complemented by significant improvements in in vivo delivery methods,which have paved the way for therapeutic application of precise gene editing tools.Effective delivery systems are critical for the success of gene therapies,and recent developments in both viral and non-viral vectors have improved the efficiency and safety of gene editing.For instance,adeno-associated viruses(AAVs)are widely used due to their high transfection efficiency and low immunogenicity,though challenges such as limited cargo capacity and potential for immune responses remain.Non-viral delivery systems,including lipid nanoparticles(LNPs),offer an alternative with lower immunogenicity and higher payload capacity,although their transfection efficiency can be lower.The therapeutic potential of these precise gene editing technologies is vast,particularly in treating genetic disorders.Preclinical studies have demonstrated the effectiveness of base editing in correcting genetic mutations responsible for diseases such as cardiomyopathy,liver disease,and hereditary hearing loss.These technologies promise to treat symptoms and potentially cure the underlying genetic causes of these conditions.Meanwhile,challenges remain,such as optimizing the safety and specificity of gene editing tools,improving delivery systems,and overcoming off-target effects,all of which are critical for their successful application in clinical settings.In summary,the continuous evolution of precise gene editing technologies,combined with advancements in delivery systems,is driving the field toward new therapeutic applications that can potentially transform the treatment of genetic disorders by targeting their root causes.展开更多
At the early stage,the transcriptome sequencing technique was used to detect the differentially expressed gene CsFK111 between vine cucumber and dwarf cucumber D0462.The gene was cloned,and bioinformatics software too...At the early stage,the transcriptome sequencing technique was used to detect the differentially expressed gene CsFK111 between vine cucumber and dwarf cucumber D0462.The gene was cloned,and bioinformatics software tools were used to analyze and predict the gene family and this gene.There were 30 members of the cucumber F-box gene family.The coding region of the cucumber CsFK111 gene was full-length 1314 bp,which encoded 437 amino acids and was predicted to be located in the nucleus.The protein encoded by this gene was a non-transmembrane protein,and the prediction of the secondary structure showed thatβ-lamellar structure and irregular crimp were dominant.A comparison of the phylogenetic tree showed that it was closest to cantaloupe and belonged to the same branch.The results provided a basis for future study on the regulation mechanism of the CsFK111 gene on cucumber dwarfing and also laid a foundation for further study of FBK family proteins.展开更多
Background Photosystem II(PSII)constitutes an intricate assembly of protein pigments,featuring extrinsic and intrinsic polypeptides within the photosynthetic membrane.The low-molecular-weight transmembrane protein Psb...Background Photosystem II(PSII)constitutes an intricate assembly of protein pigments,featuring extrinsic and intrinsic polypeptides within the photosynthetic membrane.The low-molecular-weight transmembrane protein PsbX has been identified in PSII,which is associated with the oxygen-evolving complex.The expression of PsbX gene protein is regulated by light.PsbX’s central role involves the regulation of PSII,facilitating the binding of quinone molecules to the Qb(PsbA)site,and it additionally plays a crucial role in optimizing the efficiency of photosynthesis.Despite these insights,a comprehensive understanding of the PsbX gene’s functions has remained elusive.Results In this study,we identified ten PsbX genes in Gossypium hirsutum L.The phylogenetic analysis results showed that 40 genes from nine species were classified into one clade.The resulting sequence logos exhibited substantial conservation across the N and C terminals at multiple sites among all Gossypium species.Furthermore,the ortholo-gous/paralogous,Ka/Ks ratio revealed that cotton PsbX genes subjected to positive as well as purifying selection pressure might lead to limited divergence,which resulted in the whole genome and segmental duplication.The expression patterns of GhPsbX genes exhibited variations across specific tissues,as indicated by the analysis.Moreover,the expression of GhPsbX genes could potentially be regulated in response to salt,intense light,and drought stresses.Therefore,GhPsbX genes may play a significant role in the modulation of photosynthesis under adverse abiotic conditions.Conclusion We examined the structure and function of PsbX gene family very first by using comparative genom-ics and systems biology approaches in cotton.It seems that PsbX gene family plays a vital role during the growth and development of cotton under stress conditions.Collectively,the results of this study provide basic information to unveil the molecular and physiological function of PsbX genes of cotton plants.展开更多
Objective:Toxoplasmosis is a zoonotic parasitic disease caused by Toxoplasma gondii(T.gondii),which can lead to complications such as encephalitis and ocular toxoplasmosis.The disease becomes more severe when the host...Objective:Toxoplasmosis is a zoonotic parasitic disease caused by Toxoplasma gondii(T.gondii),which can lead to complications such as encephalitis and ocular toxoplasmosis.The disease becomes more severe when the host’s immune system is compromised.Rhoptry proteins are major virulence factors that enable T.gondii to invade host cells.This study aims to construct a T.gondii rhoptry protein 41(rop41/ROP41)gene knockout strain and preliminarily investigate the biological function of rop41.Methods:Using CRISPR/Cas9 technology,a specific single-guide RNA(sgRNA)for the target gene was designed and linked to a recombinant plasmid.Homologous fragments were fused with a pyrimethamine resistance gene for selection purposes.The recombinant plasmid and the homologous fragments were electroporated into T.gondii,and PCR identification was performed after drug selection and monoclonal screening.Plaque assays were used to comprehensively assess whether rop41 affected the growth and proliferation of T.gondii in host cells.Invasion and proliferation assays were conducted to evaluate the invasion ability of the knockout strain into host cells and its intracellular proliferation capacity.The STRING database was utilized to construct a protein-protein interaction(PPI)network,and functional enrichment analysis was performed to predict the signaling pathways in which ROP41 might be involved.Results:The T.gondii rop41 gene knockout strain(RHΔku80Δrop41)was successfully constructed and stably inherited.Plaque assays showed that compared with the parental strain,the number of plaques formed by the rop41 gene knockout strain did not significantly decrease,but the reduction in plaque size was statistically significant(P<0.05).After the rop41 gene was knocked out,the invasion ability of T.gondii was reduced,but there was no statistically significant difference in its proliferation ability(P>0.05).The PPI network revealed that ROP41 was associated with other protein kinases and autophagy related proteins.Enrichment analysis indicated that proteins interacting with ROP41 may be involved in signal transduction,biosynthesis,metabolism,and autophagy-related pathways and could be components of various kinase complexes and phagocytic vesicles.Conclusion:The T.gondii RHΔku80Δrop41 strain has been successfully constructed.ROP41 primarily affects the ability of T.gondii to invade host cells and may play a role in signal transduction and autophagy-related pathways between T.gondii and the host.展开更多
This study proposes a method for uniformly revolving swarm robots to entrap multiple targets,which is based on a gene regulatory network,an adaptive decision mechanism,and an improved Vicsek-model.Using the gene regul...This study proposes a method for uniformly revolving swarm robots to entrap multiple targets,which is based on a gene regulatory network,an adaptive decision mechanism,and an improved Vicsek-model.Using the gene regulatory network method,the robots can generate entrapping patterns according to the environmental input,including the positions of the targets and obstacles.Next,an adaptive decision mechanism is proposed,allowing each robot to choose the most well-adapted capture point on the pattern,based on its environment.The robots employ an improved Vicsek-model to maneuver to the planned capture point smoothly,without colliding with other robots or obstacles.The proposed decision mechanism,combined with the improved Vicsek-model,can form a uniform entrapment shape and create a revolving effect around targets while entrapping them.This study also enables swarm robots,with an adaptive pattern formation,to entrap multiple targets in complex environments.Swarm robots can be deployed in the military field of unmanned aerial vehicles’(UAVs)entrapping multiple targets.Simulation experiments demonstrate the feasibility and superiority of the proposed gene regulatory network method.展开更多
线纹尖塘鳢(Oxyeleotris lineolata)具有典型性别生长二态性,雄鱼生长优势显著,doublesex and mab-3 related transcription factor(DMRT)家族是一个与性别决定相关的转录因子家族。基于线纹尖塘鳢性腺转录组数据,共获得2个dmrt基因的c...线纹尖塘鳢(Oxyeleotris lineolata)具有典型性别生长二态性,雄鱼生长优势显著,doublesex and mab-3 related transcription factor(DMRT)家族是一个与性别决定相关的转录因子家族。基于线纹尖塘鳢性腺转录组数据,共获得2个dmrt基因的cDNA序列,分别命名为Oxldmrt1和Oxldmrt3,并采用PCR技术扩增验证2个基因的cDNA序列。运用生物信息学方法分析2个基因序列结构特征,结果显示,Oxldmrt1和Oxldmrt3开放阅读框分别为903 bp和1363 bp,分别编码300个氨基酸和453个氨基酸;OxlDMRT1属于碱性蛋白,而OxlDMRT3属于酸性蛋白;2个基因均含有高度保守的DM结构域,OxlDMRT3还存在DMA结构域。氨基酸聚类分析显示,脊椎动物不同DMRT家族都是独立聚类,OxlDMRT1属于DMRT1家族,OxlDMRT3属于DMRT3家族,DMRT1家族最先聚类,再和DMRT3家族聚类。利用实时荧光定量PCR技术(real-time quantitative PCR,RT-qPCR)分析2个dmrt基因在雌鱼和雄鱼8个组织的表达水平,结果显示,2个基因在精巢中的表达量均显著高于其他组织(P<0.01),Oxldmrt3在脑中也有少量表达;利用RT-qPCR分析2个dmrt基因在早期不同发育时期的表达谱,显示2个基因在受精卵中的表达量均最高,Oxldmrt1在眼囊期的表达量最低,而Oxldmrt3在出膜7 d时的表达量最低。利用荧光原位杂交(fluorescencein situhybridization,FISH)对2个基因在精巢中的表达进行定位,显示2个基因在精巢中表达部位一致,均在精原细胞中有较强的表达信号。综上所述,Oxldmrt1和Oxldmrt3均在线纹尖塘鳢性腺胚胎发育阶段和精巢发育过程中起调节作用,而Oxldmrt1还可能参与胚胎后期性别决定和性别分化调控过程,Oxldmrt3还可能参与神经系统发育。本研究为线纹尖塘鳢性别决定与性别分化相关的分子机制研究提供了参考。展开更多
基金supported by the National Natural Science Foundation of China(82060418).
文摘Objective:Osteoarthritis(OA)and sarcopenia are significant health concerns in the elderly,substantially impacting their daily activities and quality of life.However,the relationship between them remains poorly understood.This study aims to uncover common biomarkers and pathways associated with both OA and sarcopenia.Methods:Gene expression profiles related to OA and sarcopenia were retrieved from the Gene Expression Omnibus(GEO)database.Differentially expressed genes(DEGs)between disease and control groups were identified using R software.Common DEGs were extracted via Venn diagram analysis.Gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses were conducted to identify biological processes and pathways associated with shared DEGs.Protein-protein interaction(PPI)networks were constructed,and candidate hub genes were ranked using the maximal clique centrality(MCC)algorithm.Further validation of hub gene expression was performed using 2 independent datasets.Receiver operating characteristic(ROC)curve analysis was used to evaluate the predictive value of key genes for OA and sarcopenia.Mouse models of OA and sarcopenia were established.Hematoxylin-eosin and Safranin O/Fast Green staining were used to validate the OA model.The sarcopenia model was validated via rotarod testing and quadriceps muscle mass measurement.Real-time reverse transcription PCR(real-time RT-PCR)was employed to assess the mRNA expression levels of candidate key genes in both models.Gene set enrichment analysis(GSEA)was conducted to identify pathways associated with the selected shared key genes in both diseases.Results:A total of 89 common DEGs were identified in the gene expression profiles of OA and sarcopenia,including 76 upregulated and 13 downregulated genes.These 89 DEGs were significantly enriched in protein digestion and absorption,the PI3K-Akt signaling pathway,and extracellular matrix-receptor interaction.PPI network analysis and MCC algorithm analysis of the 89 common DEGs identified the top 17 candidate hub genes.Based on the differential expression analysis of these 17 candidate hub genes in the validation datasets,AEBP1 and COL8A2 were ultimately selected as the common key genes for both diseases,both of which showed a significant upregulation trend in the disease groups(all P<0.05).The value of area under the curve(AUC)for AEBP1 and COL8A2 in the OA and sarcopenia datasets were all greater than 0.7,indicating that both genes have potential value in predicting OA and sarcopenia.Real-time RT-PCR results showed that the mRNA expression levels of AEBP1 and COL8A2 were significantly upregulated in the disease groups(all P<0.05),consistent with the results observed in the bioinformatics analysis.GSEA revealed that AEBP1 and COL8A2 were closely related to extracellular matrix-receptor interaction,ribosome,and oxidative phosphorylation in OA and sarcopenia.Conclusion:AEBP1 and COL8A2 have the potential to serve as common biomarkers for OA and sarcopenia.The extracellular matrix-receptor interaction pathway may represent a potential target for the prevention and treatment of both OA and sarcopenia.
基金supported by the National Natural Science Foundation(82271057)the Natural Science Foundation of Hunan Province(2023JJ30818),China。
文摘Objective:Keratoconus(KC)is a progressive corneal ectasia disorder,arising from a myriad of causes including genetic predispositions,environmental factors,biomechanical influences,and inflammatory reactions.This study aims to identify potential pathogenetic gene mutations in patients with sporadic KC in the Han Chinese population.Methods:Twenty-five patients with primary KC as well as 50 unrelated population matched healthy controls,were included in this study to identify potential pathogenic gene mutations among sporadic KC patients in the Han Chinese population.Sanger sequencing and whole-exome sequencing(WES)were used to analyze mutations in the zinc finger protein 469(ZNF469)gene.Bioinformatics analysis was conducted to explore the potential role of ZNF469 in KC pathogenesis.Results:Five novel heterozygous missense variants were identified in KC patients.Among them,2 compound heterozygous variants,c.8986G>C(p.E2996Q)with c.11765A>C(p.D3922A),and c.4423C>G(p.L1475V)with c.10633G>A(p.G3545R),were determined to be possible pathogenic factors for KC.Conclusion:Mutations in the ZNF469 gene may contribute to the development of KC in the Han Chinese population.These mutation sites may provide valuable information for future genetic screening of KC patients and their families.
文摘Objective To investigate the structural changes of rat thoracic aorta and changes in expression levels of Bmal1 and cyclins in thoracic aorta endothelial cells following heat stress.Methods Twenty male SD rats were randomized equally into control group and heat stress group.After exposure to 32℃for 2 weeks in the latter group,the rats were examined for histopathological changes and Bmal1 expression in the thoracic aorta using HE staining and immunohistochemistry.In the cell experiments,cultured rat thoracic aortic endothelial cells(RTAECs)were incubated at 40℃for 12 h with or without prior transfection with a Bmal1-specific small interfering RNA(si-Bmal1)or a negative sequence.In both rat thoracic aorta and RTAECs,the expressions of Bmal1,the cell cycle proteins CDK1,CDK4,CDK6,and cyclin B1,and apoptosis-related proteins Bax and Bcl-2 were detected using Western blotting.TUNEL staining was used to detect cell apoptosis in rat thoracic aorta,and the changes in cell cycle distribution and apoptosis in RTAECs were analyzed with flow cytometry.Results Compared with the control rats,the rats exposed to heat stress showed significantly increased blood pressures and lowered heart rate with elastic fiber disruption and increased expressions of Bmal1,cyclin B1 and CDK1 in the thoracic aorta(P<0.05).In cultured RTAECs,heat stress caused significant increase of Bmal1,cyclin B1 and CDK1 protein expression levels,which were obviously lowered in cells with prior si-Bmal1 transfection.Bmal1 knockdown also inhibited heat stress-induced increase of apoptosis in RTAECs as evidenced by decreased expression of Bax and increased expression of Bcl-2.Conclusion Heat stress upregulates Bmal1 expression and causes alterations in expressions of cyclins to trigger apoptosis of rat thoracic aorta endothelial cells,which can be partly alleviated by suppressing Bmal1 expression.
基金supported by National Natural Science Foundation of China(No.32160615).
文摘Background Verticillium dahliae,a soil-borne fungi,can cause Verticillium wilt,and seriously diminish the yield and quality of cotton.However,the pathogenic mechanism of V.dahliae is complex and not clearly understood at the moment.This study aimed to identify the high-affinity nicotinic acid transporter genes in V.dahliae.The gene expression profiles in V.dahliae following sensing of root exudates from susceptible and resistant cotton varieties were analyzed.The function of VdNAT1 in the pathogenic process of V.dahliae was studied using the tobacco rattle virus(TRV)-based host-induced gene silencing(HIGS)technique.Results Eight high-affinity nicotinic acid transporter genes were identified from V.dahliae through the bioinformatics method.Each protein contains a conserved major facilitator superfamily(MFS)domain,which belongs to the MFS superfamily.Evolutionary relationship analysis revealed that all 8 genes belong to the anion:cation symporter(ACS)subfamily.All proteins have transmembrane domains,ranging from 7 to 12.The expression levels of most VdNAT genes were significantly increased after induction by root exudates from susceptible cotton varieties.Silencing VdNAT1 gene by HIGS significantly inhibited the accumulation of fungal biomass in cotton plants,and alleviated the disease symptoms of cotton.Conclusions Eight VdNAT genes were identified from V.dahliae,and most VdNAT genes was up-regulated after induced by root exudates from susceptible cotton variety.In addition,VdNAT1 is required for the pathogenicity of V.dahliae.Overall,these findings will facilitate the pathogenic molecular mechanism of V.dahliae and provide candidate genes.
文摘The advent of gene editing represents one of the most transformative breakthroughs in life science,making genome manipulation more accessible than ever before.While traditional CRISPR/Cas-based gene editing,which involves double-strand DNA breaks(DSBs),excels at gene disruption,it is less effective for accurate gene modification.The limitation arises because DSBs are primarily repaired via non-homologous end joining(NHEJ),which tends to introduce indels at the break site.While homology directed repair(HDR)can achieve precise editing when a donor DNA template is provided,the reliance on DSBs often results in unintended genome damage.HDR is restricted to specific cell cycle phases,limiting its application.Currently,gene editing has evolved to unprecedented levels of precision without relying on DSB and HDR.The development of innovative systems,such as base editing,prime editing,and CRISPR-associated transposases(CASTs),now allow for precise editing ranging from single nucleotides to large DNA fragments.Base editors(BEs)enable the direct conversion of one nucleotide to another,and prime editors(PEs)further expand gene editing capabilities by allowing for the insertion,deletion,or alteration of small DNA fragments.The CAST system,a recent innovation,allows for the precise insertion of large DNA fragments at specific genomic locations.In recent years,the optimization of these precise gene editing tools has led to significant improvements in editing efficiency,specificity,and versatility,with advancements such as the creation of base editors for nucleotide transversions,enhanced prime editing systems for more efficient and precise modifications,and refined CAST systems for targeted large DNA insertions,expanding the range of applications for these tools.Concurrently,these advances are complemented by significant improvements in in vivo delivery methods,which have paved the way for therapeutic application of precise gene editing tools.Effective delivery systems are critical for the success of gene therapies,and recent developments in both viral and non-viral vectors have improved the efficiency and safety of gene editing.For instance,adeno-associated viruses(AAVs)are widely used due to their high transfection efficiency and low immunogenicity,though challenges such as limited cargo capacity and potential for immune responses remain.Non-viral delivery systems,including lipid nanoparticles(LNPs),offer an alternative with lower immunogenicity and higher payload capacity,although their transfection efficiency can be lower.The therapeutic potential of these precise gene editing technologies is vast,particularly in treating genetic disorders.Preclinical studies have demonstrated the effectiveness of base editing in correcting genetic mutations responsible for diseases such as cardiomyopathy,liver disease,and hereditary hearing loss.These technologies promise to treat symptoms and potentially cure the underlying genetic causes of these conditions.Meanwhile,challenges remain,such as optimizing the safety and specificity of gene editing tools,improving delivery systems,and overcoming off-target effects,all of which are critical for their successful application in clinical settings.In summary,the continuous evolution of precise gene editing technologies,combined with advancements in delivery systems,is driving the field toward new therapeutic applications that can potentially transform the treatment of genetic disorders by targeting their root causes.
基金Supported by the National Natural Science Foundation of China(32272724)the National Science Foundation of Heilongjiang Province,China(LH2019C033)。
文摘At the early stage,the transcriptome sequencing technique was used to detect the differentially expressed gene CsFK111 between vine cucumber and dwarf cucumber D0462.The gene was cloned,and bioinformatics software tools were used to analyze and predict the gene family and this gene.There were 30 members of the cucumber F-box gene family.The coding region of the cucumber CsFK111 gene was full-length 1314 bp,which encoded 437 amino acids and was predicted to be located in the nucleus.The protein encoded by this gene was a non-transmembrane protein,and the prediction of the secondary structure showed thatβ-lamellar structure and irregular crimp were dominant.A comparison of the phylogenetic tree showed that it was closest to cantaloupe and belonged to the same branch.The results provided a basis for future study on the regulation mechanism of the CsFK111 gene on cucumber dwarfing and also laid a foundation for further study of FBK family proteins.
基金supported by National Natural Science Foundation of China(32060466)Chinese Academy of Agricultural Sciences。
文摘Background Photosystem II(PSII)constitutes an intricate assembly of protein pigments,featuring extrinsic and intrinsic polypeptides within the photosynthetic membrane.The low-molecular-weight transmembrane protein PsbX has been identified in PSII,which is associated with the oxygen-evolving complex.The expression of PsbX gene protein is regulated by light.PsbX’s central role involves the regulation of PSII,facilitating the binding of quinone molecules to the Qb(PsbA)site,and it additionally plays a crucial role in optimizing the efficiency of photosynthesis.Despite these insights,a comprehensive understanding of the PsbX gene’s functions has remained elusive.Results In this study,we identified ten PsbX genes in Gossypium hirsutum L.The phylogenetic analysis results showed that 40 genes from nine species were classified into one clade.The resulting sequence logos exhibited substantial conservation across the N and C terminals at multiple sites among all Gossypium species.Furthermore,the ortholo-gous/paralogous,Ka/Ks ratio revealed that cotton PsbX genes subjected to positive as well as purifying selection pressure might lead to limited divergence,which resulted in the whole genome and segmental duplication.The expression patterns of GhPsbX genes exhibited variations across specific tissues,as indicated by the analysis.Moreover,the expression of GhPsbX genes could potentially be regulated in response to salt,intense light,and drought stresses.Therefore,GhPsbX genes may play a significant role in the modulation of photosynthesis under adverse abiotic conditions.Conclusion We examined the structure and function of PsbX gene family very first by using comparative genom-ics and systems biology approaches in cotton.It seems that PsbX gene family plays a vital role during the growth and development of cotton under stress conditions.Collectively,the results of this study provide basic information to unveil the molecular and physiological function of PsbX genes of cotton plants.
基金supported by the National Natural Science Foundation of China(32170510)the Innovation Training Program of Central South University(20240026020055),China.
文摘Objective:Toxoplasmosis is a zoonotic parasitic disease caused by Toxoplasma gondii(T.gondii),which can lead to complications such as encephalitis and ocular toxoplasmosis.The disease becomes more severe when the host’s immune system is compromised.Rhoptry proteins are major virulence factors that enable T.gondii to invade host cells.This study aims to construct a T.gondii rhoptry protein 41(rop41/ROP41)gene knockout strain and preliminarily investigate the biological function of rop41.Methods:Using CRISPR/Cas9 technology,a specific single-guide RNA(sgRNA)for the target gene was designed and linked to a recombinant plasmid.Homologous fragments were fused with a pyrimethamine resistance gene for selection purposes.The recombinant plasmid and the homologous fragments were electroporated into T.gondii,and PCR identification was performed after drug selection and monoclonal screening.Plaque assays were used to comprehensively assess whether rop41 affected the growth and proliferation of T.gondii in host cells.Invasion and proliferation assays were conducted to evaluate the invasion ability of the knockout strain into host cells and its intracellular proliferation capacity.The STRING database was utilized to construct a protein-protein interaction(PPI)network,and functional enrichment analysis was performed to predict the signaling pathways in which ROP41 might be involved.Results:The T.gondii rop41 gene knockout strain(RHΔku80Δrop41)was successfully constructed and stably inherited.Plaque assays showed that compared with the parental strain,the number of plaques formed by the rop41 gene knockout strain did not significantly decrease,but the reduction in plaque size was statistically significant(P<0.05).After the rop41 gene was knocked out,the invasion ability of T.gondii was reduced,but there was no statistically significant difference in its proliferation ability(P>0.05).The PPI network revealed that ROP41 was associated with other protein kinases and autophagy related proteins.Enrichment analysis indicated that proteins interacting with ROP41 may be involved in signal transduction,biosynthesis,metabolism,and autophagy-related pathways and could be components of various kinase complexes and phagocytic vesicles.Conclusion:The T.gondii RHΔku80Δrop41 strain has been successfully constructed.ROP41 primarily affects the ability of T.gondii to invade host cells and may play a role in signal transduction and autophagy-related pathways between T.gondii and the host.
基金funded by the National Natural Science Foundation of China(62176147)the Science and Technology Planning Project of Guangdong Province of China,the State Key Lab of Digital Manufacturing Equipment and Technology(DMETKF2019020)the National Defense Technology Innovation Special Zone Project(193-A14-226-01-01)。
文摘This study proposes a method for uniformly revolving swarm robots to entrap multiple targets,which is based on a gene regulatory network,an adaptive decision mechanism,and an improved Vicsek-model.Using the gene regulatory network method,the robots can generate entrapping patterns according to the environmental input,including the positions of the targets and obstacles.Next,an adaptive decision mechanism is proposed,allowing each robot to choose the most well-adapted capture point on the pattern,based on its environment.The robots employ an improved Vicsek-model to maneuver to the planned capture point smoothly,without colliding with other robots or obstacles.The proposed decision mechanism,combined with the improved Vicsek-model,can form a uniform entrapment shape and create a revolving effect around targets while entrapping them.This study also enables swarm robots,with an adaptive pattern formation,to entrap multiple targets in complex environments.Swarm robots can be deployed in the military field of unmanned aerial vehicles’(UAVs)entrapping multiple targets.Simulation experiments demonstrate the feasibility and superiority of the proposed gene regulatory network method.
