Background:Alterations in splicing factors contribute to aberrant alternative splicing(AS),which subsequently promotes tumor progression.The splicing factor polypyrimidine tract binding protein 1(PTBP1)has been shown ...Background:Alterations in splicing factors contribute to aberrant alternative splicing(AS),which subsequently promotes tumor progression.The splicing factor polypyrimidine tract binding protein 1(PTBP1)has been shown to facilitate cancer progression by modulating oncogenic variants.However,its specific role and underlying mechanisms in hepatocellular carcinoma(HCC)remain to be elucidated.Methods:PTBP1 expression was evaluated in HCC tissues and cell lines.Subsequently,cells were transfected with vectors designed for PTBP1 overexpression or downregulation.The biological function of PTBP1 was assessed in vitro and in vivo using MTS assays,colony formation assays,transwell assays,xenograft formation,tail vein injection,and orthotopic models.Transcriptome analysis was conducted to elucidate the underlying molecular mechanisms.Results:Our findings demonstrated that PTBP1 exhibited elevated expression in HCC cell lines and tissues.Furthermore,its expression positively correlated with overall and disease-free survival rates,as well as tumor grade and stage.PTBP1 knockdown reduced HCC cell proliferation,migration,and invasion in vitro and suppressed hepatocarcinoma xenograft growth and infiltration in vivo.RNA sequencing(RNA-Seq)analysis identified the AS events associated with PTBP1.PTBP1 functionally enhanced cell proliferation,invasion,and migration by modulating the AS of the microtubule-associated protein tau(MAPT)gene and promoting oncogene expression.Notably,the dysregulation of MAPT splicing coincided with increased PTBP1 expression in HCC.Conclusions:PTBP1-guided AS of the MAPT gene enhances tumorigenicity in HCC through activation of the MAPK/ERK pathways.展开更多
OCT4 is one of the key transcription factors in maintaining the pluripotency and self-renewal of embryonic stem （ES） cells.Human OCT4 can generate two isoforms OCT4A and OCT4B by alternative splicing.OCT4B1 is a rec...OCT4 is one of the key transcription factors in maintaining the pluripotency and self-renewal of embryonic stem （ES） cells.Human OCT4 can generate two isoforms OCT4A and OCT4B by alternative splicing.OCT4B1 is a recently discovered novel OCT4 spliced variant,which has been considered as a putative marker of stemness.Compared with the OCT4B mRNA,OCT4B1 mRNA is generated by retaining intron 2 as a cryptic exon which contains a TGA stop codon in it.As a result,the protein product of OCT4B1 mRNA could be truncated.Interestingly,we present here that OCT4B1 can indirectly produce the same protein products as OCT4B.We have demonstrated that OCT4B1 mRNA can be spliced into OCT4B mRNA,and encode three protein isoforms.The splicing of OCT4B1 mRNA into OCT4B mRNA can be remarkably inhibited by the mutation of the classical splicing site.Our result suggests that OCT4B mRNA may originate from OCT4B1 mRNA by alternative splicing.展开更多
Developing effective irrigation and drainage strategies to improve the quality of saline-alkali soil is vital for enhancing agricultural production and increasing economic returns. In this study, we explored how irrig...Developing effective irrigation and drainage strategies to improve the quality of saline-alkali soil is vital for enhancing agricultural production and increasing economic returns. In this study, we explored how irrigation and drainage modes (flood irrigation, drip irrigation, and sub-surface pipe drainage under drip irrigation) improve the saline-alkali soil in Xinjiang, China. We aimed to study the transport characteristics of soil water and salt under different irrigation and drainage modes, and analyze the effects of the combination of irrigation and drainage on soil salt leaching, as well as its impacts on the growth of oil sunflower. Our results show that sub-surface pipe drainage under drip irrigation significantly reduced the soil salt content and soil water content at the 0–200 cm soil depth. Under sub-surface pipe drainage combined with drip irrigation, the mean soil salt content was reduced to below 10 g/kg after the second irrigation, and the soil salt content decreased as sub-surface pipe distance decreased. The mean soil salt content of flood irrigation exceeded 25 g/kg, and the mean soil desalination efficiency was 3.28%, which was lower than that of drip irrigation. The mean soil desalination rate under drip irrigation and sub-surface pipe drainage under drip irrigation was 19.30% and 58.12%, respectively. After sub-surface drainage regulation under drip irrigation, the germination percentage of oil sunflower seedlings was increased to more than 50%, which further confirmed that combined drip irrigation and sub-surface pipe drainage is very effective in improving the quality of saline-alkali soil and increasing the productivity of agricultural crops.展开更多
The removal efficiency of Cu and Zn from swine wastewater was evaluated as effected by three variables: the hydraulic retention time (HRT) (24, 48, 72 and 96 hours), two different plant species (Typha domingensis Pers...The removal efficiency of Cu and Zn from swine wastewater was evaluated as effected by three variables: the hydraulic retention time (HRT) (24, 48, 72 and 96 hours), two different plant species (Typha domingensis Pers. and Eleocharis cellulosa) and two different sizes of filter media (5 and 15 mm) using a horizontal sub-surface flow constructed wetland. From the results, a significant difference was observed in the removal efficiency of Cu and Zn with respect to different hydraulic retention times. The best results were obtained in the HRT of 96 hours for Zn where 96% removal of Zn with Typha domingensis Pers. specie with gravel of 15 mm (experimental unit 6) was achieved. For Cu, at 72 hours of HRT, the efficiency was nearly 100% in five of the six study units (1, 2, 3, 5 and 6). In contrast, in experimental unit 4 with gravel of 15 mm and without plants, only 86% Cu removal was achieved.展开更多
An Arctic Ocean eddy in sub-surface layer is analyzed in this paper by use of temperature, salinity and current profiles data obtained at an ice camp in the Canada Basin during the second Chinese Arctic Expedition in ...An Arctic Ocean eddy in sub-surface layer is analyzed in this paper by use of temperature, salinity and current profiles data obtained at an ice camp in the Canada Basin during the second Chinese Arctic Expedition in summer of 2003. In the vertical temperature section, the eddy shows itself as an isolated cold water block at depth of 60 m with a minimum temperature of - 1.5℃, about 0.5℃ colder than the ambient water. Isopycnals in the eddy form a pattern of convex, which indicates the eddy is anticyclonic. Although maximum velocity near 0.4 m s^-1 occurs in the current records observed synchronously, the current pattern is far away from a typical eddy. By further analysis, inertial frequency osci/lations with amplitudes comparable with the eddy velocity are found in the sub-surface layer currents. After filter the inertial current and mean current, an axisymmetric current pattern of an eddy with maximum velocity radius of 5 km is obtained. The analysis of the T-S characteristics of the eddy core water and its ambient waters supports the conclusion that the eddy was formed on the Chukchi Shelf and migrated northeastward into the northern Canada Basin.展开更多
The shaping form of an untwisted yarn-end for the air splicer is analyzed and then a cosine curve form which will have practically good splicing effect is proposed. The yarn motion in the splicing nozzle by applying h...The shaping form of an untwisted yarn-end for the air splicer is analyzed and then a cosine curve form which will have practically good splicing effect is proposed. The yarn motion in the splicing nozzle by applying hydrodynamics and the splicing principle of the air splicer are also studied.展开更多
The aim of the present study is to improve the capabilities and precision of a recently introduced Sea Surface Acoustic Simulator(SSAS) developed based on optimization of the Helmholtz–Kirchhoff–Fresnel(HKF) method....The aim of the present study is to improve the capabilities and precision of a recently introduced Sea Surface Acoustic Simulator(SSAS) developed based on optimization of the Helmholtz–Kirchhoff–Fresnel(HKF) method. The improved acoustic simulator, hereby known as the Modified SSAS(MSSAS), is capable of determining sound scattering from the sea surface and includes an extended Hall–Novarini model and optimized HKF method. The extended Hall–Novarini model is used for considering the effects of sub-surface bubbles over a wider range of radii of sub-surface bubbles compared to the previous SSAS version. Furthermore, MSSAS has the capability of making a three-dimensional simulation of scattered sound from the rough bubbly sea surface with less error than that of the Critical Sea Tests(CST) experiments. Also, it presents scattered pressure levels from the rough bubbly sea surface based on various incident angles of sound. Wind speed, frequency, incident angle, and pressure level of the sound source are considered as input data, and scattered pressure levels and scattering coefficients are provided. Finally, different parametric studies were conducted on wind speeds, frequencies, and incident angles to indicate that MSSAS is quite capable of simulating sound scattering from the rough bubbly sea surface, according to the scattering mechanisms determined by Ogden and Erskine. Therefore, it is concluded that MSSAS is valid for both scattering mechanisms and the transition region between them that are defined by Ogden and Erskine.展开更多
RNA splicing normally generates stable splice- junction sequences in viruses that are important in the context of virus mimicry. Potential variability in envelop proteins may occur with point-mutations inducing crypti...RNA splicing normally generates stable splice- junction sequences in viruses that are important in the context of virus mimicry. Potential variability in envelop proteins may occur with point-mutations inducing cryptic splice-junctions, which would remain unrecognized by T-memory cells of higher organisms in vaccine trials. Such aberrant splice- junctions result from evolution-specific non-conser- vation of actual splice-junction sites due to mutations;as such, locations of splice-junctions in a test DNA sequence could only be imprecisely specified. Such impreciseness of splice-junction locations (or cryptic sites) in a sequence is evaluated in this study via “noisy” attributes (with associated stochastics) to the mutated subspace;and, relevant fuzzy considerations are invoked with membership attributes expressed in terms of a spatial signal-to-noise ratio (SSNR). That is, SSNR adopted as a membership function expresses the belongingness of a site-region to exon/intron subspaces. An illustrative example with actual (Dengue 1 viral) DNA data is furnished demonstrating the pursuit developed in predicting aberrant splice-junctions at cryptic sites in the test sequence.展开更多
This study aims at identifying possible correlations between shapes, types of geophysical anomalies and borehole productivity according to geological and hydrogeotogical contexts. The methodology adopted was a two-pro...This study aims at identifying possible correlations between shapes, types of geophysical anomalies and borehole productivity according to geological and hydrogeotogical contexts. The methodology adopted was a two-pronged one--the first step sought to： （1） interpret the electrical resistivity values from horizontal profiling and vertical electrical sounding implemented in Seno province that preceded the drilling of 513 boreholes; （ii） interpret data from pumping tests carried out on boreholes having a discharge superior to 1 m3/h （＂positive borehole＂） by using Cooper-Jacob＇s method. In the second step, according to geology, authors tried to identify possible correlations between each of the qualitative geophysical parameters： ~shape of anomaly〉〉, tttype of anomaly〉〉 and ~〈type curve〉〉 on the one hand, and hydrogeological parameters such as discharge, alteration thickness, transmissivity and saturated level on the other. The results of this study have shown that the chances of having a positive borehole in Seno province are higher when the type of anomaly is TCC （80%）, shape of anomaly is ＂W＂ and when type curve is ＂H＂ （80%） for all geological formations. Granitic formations are those that record higher discharges while schists record high transmissivity values.展开更多
Maize being sub-tropical crop is sensitive to water deficit during the early growth stages;particularly clay-rich soil,due to the compaction of the soil.It is well-documented that potential sub-surface drip irrigation...Maize being sub-tropical crop is sensitive to water deficit during the early growth stages;particularly clay-rich soil,due to the compaction of the soil.It is well-documented that potential sub-surface drip irrigation(SDI)(Full irrigation;SDIFull(100%field capacity(FC)),Deficit irrigation;SDIDeficit(70%FC))improves water use efficiency,which leads to increased crop productivity;since it has a constraint that SDI excludes soil air around the root-zone during irrigation events,which alter the root function and crop performance.Additionally,in clay-rich soils,the root system of plants generally suffers the limitation of oxygen,particularly the temporal hypoxia,and occasionally from root anoxia;while SDI system accomplishes with the aerating stream of irrigation in the rhizosphere could provide oxygen root environment.The oxygen can be introduced into the irrigation stream of SDI through two ways:the venturi principle,or by using solutions of hydrogen peroxide through the air injection system.Therefore,the application of hydrogen peroxide(H_(2)O_(2);HP)can mitigate the adverse effect of soil compactness and also lead to improving the growth,yield and yield attributes of maize in clay-rich soil.Considering the burning issue,a field study was conducted in consecutive two seasons of 2017 and 2018;where hybrid maize was cultivated as a second crop,to evaluate the effect of liquid-injection of H_(2)O_(2)(HP)into the irrigation stream of SDI on the performance of maize in a clay-rich soil field of Adana,Turkey.When soil water content decreased in 50%of avail-able water,irrigation was performed.The amount of water applied to reach the soil water content to the field capacity is SDIFull(100%FC)and 70%FC of this water is SDIDeficit(70%FC).In the irrigation program,hydrogen peroxide(HP)was applied at intervals of 7 days on average according to available water with and without HP:SDIFull(100%FC)+0 ppm HP with full SDI irrigation;SDIFull(100%FC)+250 ppm HP with deficit SDI irrigation;SDIDeficit(70%FC)+0 ppm HP,SDIDeficit(70%FC)+250 ppm HP and SDIDeficit(70%FC)+500 ppm HP.Deficit irrigation(SDIDeficit(70%FC))program was started from tasseling stage and continued up to the physiological maturity stage with sub-soil drip irrigation.H_(2)O_(2) was applied 3 times during the growing season.Two years’results revealed that the liquid-injection of H_(2)O_(2) into the irrigation stream of SDI improved the growth and yield-related attributes and grain yield of maize.Based on the obtained results,during the extreme climatic condition in the year 2017,SDIFull(100%FC)+250 ppm HP was more effective than SDIFull(100%FC)+0 ppm HP on all traits for relative to full irrigation.While,during the favourable climatic condition in the 2018 season,SDIFull(100%FC)+250 ppm HP was more effective than full irrigation with SDIFull(100%FC)+0 ppm HP for the grain yield,grains,and SPAD value.Accordingly,the most effective treatment was SDIFull(100%FC)+250 ppm HP,as it gave the highest growth and yield-related attributes and grain yield of maize followed by SDIDeficit(70%FC)+250 ppm HP.Therefore,SDIFull with 250 ppm H_(2)O_(2) using as liquid-injection may be recommended to mitigate the adverse effect of soil compactness particularly water-deficit stress in clay-rich soil for the sustainability of maize production.展开更多
The formation of root system architecture(RSA)plays a crucial role in plant growth.OsDRO1 is known to have a function in controlling RSA in rice,however,the role of potato StDRO2,a homolog of rice OsDRO1,in root growt...The formation of root system architecture(RSA)plays a crucial role in plant growth.OsDRO1 is known to have a function in controlling RSA in rice,however,the role of potato StDRO2,a homolog of rice OsDRO1,in root growth remains unclear.In this study,we obtained potato dro2 mutant lines by Clustered Regularly Interspaced Short Palindromic Repeats-CRISPR-Associated 9(CRISPR/Cas9)-mediated genome editing system.The mutant lines were generated from a splicing defect of the StDRO2 intron 1,which causes a nonsense mutation in StDRO2.Furthermore,the secondary structure of StDRO2 mRNA analyzed with RNAfold Web Server was altered in the dro2 mutant.Mutation of StDRO2 conveys potato adaptation through changing the RSA via alteration of auxin transport under drought stress.The potato dro2 lines showed higher plant height,longer root length,smaller root growth angle and increased tuber weight than the wild-type.The alteration of RSA was associated with a disturbance of IAA distribution in the dro2 mutant,and the levels of StPIN7 and StPIN10 detected by using real-time PCR were up-regulated in the roots of potato dro2 lines grown under drought stress.Moreover,the microRNAs(miRNAs)PmiREN024536 and PmiREN024486 targeted the StDRO2 gene,and auxin positively and negatively regulated the expression of StDRO2 and the miRNAs PmiREN024536 and PmiREN024486,respectively,in the potato roots.Our data shows that a regulatory network involving auxin,StDRO2,PmiREN024536 and PmiREN024486 can control RSA to convey potato fitness under drought stress.展开更多
Pentatricopeptide repeat(PPR)proteins are a large group of eukaryote-specific RNA-binding proteins that play pivotal roles in plant organelle gene expression.Here,we report the function of PPR21 in mitochondrial intro...Pentatricopeptide repeat(PPR)proteins are a large group of eukaryote-specific RNA-binding proteins that play pivotal roles in plant organelle gene expression.Here,we report the function of PPR21 in mitochondrial intron splicing and its role in maize kernel development.PPR21 is a typical P-type PPR protein targeted to mitochondria.The ppr21 mutants are arrested in embryogenesis and endosperm development,leading to embryo lethality.Null mutations of PPR21 reduce the splicing efficiency of nad2 intron 1,2,and 4 and impair the assembly and activity of mitochondrial complex I.Previous studies show that the P-type PPR protein EMP12 is required for the splicing of identical introns.However,our protein interaction analyses reveal that PPR21 does not interact with EMP12.Instead,both PPR21 and EMP12 interact with the small MutS-related(SMR)domain-containing PPR protein 1(PPR-SMR1)and the short P-type PPR protein 2(SPR2).PPR-SMR1 interacts with SPR2,and both proteins are required for the splicing of many introns in mitochondria,including nad2 intron 1,2,and 4.These results suggest that a PPR21-(PPR-SMR1/SPR2)-EMP12 complex is involved in the splicing of nad2 introns in maize mitochondria.展开更多
In this study,we examine the problem of sliced inverse regression(SIR),a widely used method for sufficient dimension reduction(SDR).It was designed to find reduced-dimensional versions of multivariate predictors by re...In this study,we examine the problem of sliced inverse regression(SIR),a widely used method for sufficient dimension reduction(SDR).It was designed to find reduced-dimensional versions of multivariate predictors by replacing them with a minimally adequate collection of their linear combinations without loss of information.Recently,regularization methods have been proposed in SIR to incorporate a sparse structure of predictors for better interpretability.However,existing methods consider convex relaxation to bypass the sparsity constraint,which may not lead to the best subset,and particularly tends to include irrelevant variables when predictors are correlated.In this study,we approach sparse SIR as a nonconvex optimization problem and directly tackle the sparsity constraint by establishing the optimal conditions and iteratively solving them by means of the splicing technique.Without employing convex relaxation on the sparsity constraint and the orthogonal constraint,our algorithm exhibits superior empirical merits,as evidenced by extensive numerical studies.Computationally,our algorithm is much faster than the relaxed approach for the natural sparse SIR estimator.Statistically,our algorithm surpasses existing methods in terms of accuracy for central subspace estimation and best subset selection and sustains high performance even with correlated predictors.展开更多
目的探讨虚拟仿真结合改良自我导向、问题导向、终身学习、综合临床案例练习(selfdirected,problem-oriented,lifelong learning,integrated clinical case exercise,SPLICE)教学法对临床医学专硕生批判性思维能力的影响。方法选取2024...目的探讨虚拟仿真结合改良自我导向、问题导向、终身学习、综合临床案例练习(selfdirected,problem-oriented,lifelong learning,integrated clinical case exercise,SPLICE)教学法对临床医学专硕生批判性思维能力的影响。方法选取2024年1—6月在锦州医科大学附属第一医院皮肤科进行住院医师规范化培训(简称“住培”)的64名2023级临床医学专硕生为研究对象。采用随机分组法将其分为试验组(n=32)和对照组(n=32)。试验组采用虚拟仿真结合改良SPLICE教学,对照组采用传统教学方式。比较两组学生批判性思维能力、学业自我效能感、理论考核和临床操作技能考核成绩。结果试验组医学专硕生的批判性思维能力、学业自我效能感、理论考核和临床操作技能考核成绩均高于对照组,差异具有统计学意义(均P<0.001)。结论虚拟仿真结合改良SPLICE教学法有助于提升医学专硕生的批判性思维能力与学业自我效能感,有利于其更好地掌握专业理论和临床操作技能,值得在临床教学中进一步推广应用。展开更多
ObjectivesThe PTPRQ gene is essential for preserving the structure and function of stereocilia in inner ear.However,research on splicing mutations within this gene is limited.This study aims to investigate novel splic...ObjectivesThe PTPRQ gene is essential for preserving the structure and function of stereocilia in inner ear.However,research on splicing mutations within this gene is limited.This study aims to investigate novel splicing mutations in PTPRQ,clarify their molecular mechanisms,and provide new insights into the genetic factors associated with hearing loss,ultimately enhancing diagnostic accuracy.MethodClinical data and peripheral blood samples were obtained from members of a family with congenital hearing loss.Variants were identified through high-throughput sequencing and confirmed by Sanger sequencing to ensure genealogical co-segregation.The splicing effects of PTPRQ variants were evaluated using bioinformatics tools and minigene assays.ResultsWe used whole exome sequencing to identify novel double compound heterozygous splice-altering variants(c.5426+1 G>A and c.6603-3 T>G)in the PTPRQ gene with DFNB84A.We molecularly characterized these variants,and they were found to co-segregate with the disease within the family.Minigene assays and Sanger sequencing confirmed that the c.6603-3 T>G variant caused exon 43 skipping,resulting in a frameshift mutation(p.Ser2201ArgfsTer112).Further bioinformatic analysis supported these findings.ConclusionsThis study identifies a novel compound heterozygous splicing variant in the PTPRQ gene in a Chinese family with DFNB84A,expanding the known spectrum of PTPRQ mutations.These findings enhance the understanding of PTPRQ-related hearing loss and may aid in early diagnosis,prevention,and therapeutic strategies.展开更多
Caspases,which play key roles in cell apoptosis,undergo alternative splicing to form different splicing variants that can regulate the apoptotic process.Lepidopteran insect caspases undergo alternative splicing,althou...Caspases,which play key roles in cell apoptosis,undergo alternative splicing to form different splicing variants that can regulate the apoptotic process.Lepidopteran insect caspases undergo alternative splicing,although the functions of their splicing variants are still unclear.The Spodoptera exigua caspase-5(SeCaspase-5)gene was cloned and found to produce four different splicing variants with different gene sequences and protein functional domains,which were named SeCaspase-5a,SeCaspase-5b,SeCaspase-5c and SeCaspase-5d.Overexpression of these variants in S.exigua cells(Se-3)showed that SeCaspase-5a had a proapoptotic function,whereas SeCaspase-5b,SeCaspase-5c and SeCaspase-5d did not.Semi-qPCR analysis revealed that the expression of the SeCaspase-5 variants significantly differed during Autographa californica multiple nucleopolyhedrovirus(AcMNPV)infection.Furthermore,the SeCaspase-5 variants were constructed into the AcMNPV bacmid and transfected into Se-3 cells,which revealed that SeCaspase-5a promoted cell apoptosis and reduced virus production,whereas SeCaspase-5b,SeCaspase-5c and SeCaspase-5d did not promote cell apoptosis but instead increased virus production.Moreover,an analysis of the interactions between the SeCaspase-5 variants revealed that SeCaspase-5a directly interacted with SeCaspase-5b,SeCaspase-5c and SeCaspase-5d.Coexpression of these variants in Se-3 cells also revealed that SeCaspase-5b,SeCaspase-5c and SeCaspase-5d inhibited the proapoptotic function of SeCaspase-5a,resulting in a reduction in the percentage of apoptotic cells by about 20%.These results indicate that SeCaspase-5 undergoes alternative splicing and is involved in regulating the apoptosis induced by baculovirus infection.These findings increase our understanding of the functions of lepidopteran insect caspases and provide new insights into the mechanism of host-cell apoptosis induced by baculoviruses.展开更多
Heat stress is a major threat to maize(Zea mays L.)production worldwide.Heat shock transcription factors(HSFs)play vital roles in plant responses to heat stress.However,the molecular and genetic mechanisms underlying ...Heat stress is a major threat to maize(Zea mays L.)production worldwide.Heat shock transcription factors(HSFs)play vital roles in plant responses to heat stress.However,the molecular and genetic mechanisms underlying HSF-meditated thermotolerance in maize remain largely unexplored.In this study,we demonstrate that the alternative splicing of ZmHsf23 modulates heat stress tolerance in maize.Hsf23 produced two functional transcripts,Hsf23L and Hsf23S,which differ by the presence of a cryptic mini-exon in Hsf23L that is spliced out in Hsf23S.Both transcripts were strongly induced by heat stress.Mutants lacking Hsf23L alone(hsf23l)or both Hsf23L and Hsf23S(hsf23l23s)exhibited increased susceptibility to heat stress,whereas overexpression of Hsf23S enhanced heat stress tolerance in maize.Subsequently,we found that Hsf23S positively regulates heat stress tolerance by directly activating the transcription of three sHSP genes(Hsp16.9,Hsp17.2,and Hsp18a)and TIL1 gene.In addition,Hsf23L physically interacted with Hsf23S and enhanced the transcriptional activation of Hsf23S on the sHSPs and TIL1 promoters.Notably,genetic analysis suggested that co-overexpression of Hsf23L and Hsf23S further improves heat tolerance of the transgenic plants.Taken together,these results reveal two splicing variants of ZmHsf23 cooperatively regulate maize heat tolerance,thus highlighting potential value of ZmHsf23 in breeding heat-tolerant maize varieties.展开更多
Background:Glioblastoma(GBM)is one of the most malignant types of central nervous system tumors.Oxygen deprivation in the tumor microenvironment is thought to be an important factor in promoting GBM progression.Howeve...Background:Glioblastoma(GBM)is one of the most malignant types of central nervous system tumors.Oxygen deprivation in the tumor microenvironment is thought to be an important factor in promoting GBM progression.However,the mechanisms of hypoxia-promoted tumor progression remain elusive.Methods:Alternative splicing of diacylglycerol kinase gamma(DGKG)-Δexon13 was amplified and verified by PCR-Sanger sequencing.The functions of DGKG and DGKG-Δexon13 were analyzed by Cell counting kit-8(CCK-8),Transwell,Matrigeltranswell experiments,and in vivo orthotropic GBM animal models.Transcriptome analyses were done to find out the regulated genes.Results:In this study,we found that a new transcript DGKG-Δexon13 was generated in GBM under hypoxia via alternative splicing.Moreover,the results of CCK-8,Transwell,and Matrigel-transwell experiments showed that the proliferation,migration,and invasion abilities of U87-MG and T98G were decreased after DGKG knockdown.Compared to wild-type DGKG,DGKG-Δexon13 overexpression significantly promoted cellular proliferation,migration,and invasion abilities in GBM.Furthermore,in vivo,orthotropic GBM animal models analysis showed that the tumor volumes were much smaller in the DGKG knockdown group.However,the tumor sizes in the DGKG and DGKG-Δexon13 rescue groups were restored,especially in the DGKG-Δexon13 group.Transcriptome analysis revealed that MORC1,KLHDC7B,ATP1A2,INHBE,TMEM119,and FGD3 were altered significantly when DGKG was knocked down.IL-16,CCN2,and EFNB3 were specifically regulated by DGKG-Δexon13.Conclusions:Our study found that hypoxia-induced alternative splicing transcript DGKG-Δexon13 promotes GBM proliferation and infiltration,which might provide a new potential target for the clinical treatment and diagnosis of GBM.展开更多
Starch biosynthesis is a complex process that relies on the coordinated action of multiple enzymes.Resistant starch is not digested in the small intestine,thus preventing a rapid rise in the glycemic index.Starch synt...Starch biosynthesis is a complex process that relies on the coordinated action of multiple enzymes.Resistant starch is not digested in the small intestine,thus preventing a rapid rise in the glycemic index.Starch synthase 2a(SS2a)is a key enzyme in amylopectin biosynthesis that has significant effects on starch structure and properties.In this study,we identified an ss2a null mutant(M3-1413)with a single base mutation from an ethyl methane sulfonate(EMS)-mutagenized population of barley.The mutation was located at the 3'end of the first intron of the RNA splicing receptor(AG)site,and resulted in abnormal RNA splicing and two abnormal transcripts of ss2a,which caused the inactivation of the SS2a gene.The starch structure and properties were significantly altered in the mutant,with M3-1413 containing lower total starch and higher amylose and resistant starch levels.This study sheds light on the effect of barley ss2a null mutations on starch properties and will help to guide new applications of barley starch in the development of nutritious food products.展开更多
基金supported by the National Natural Science Foundation of China(Nos.81972771,82173062)the Key Areas Project of Education Department of Guangdong Province(No.2021ZDZX2017)+3 种基金the Tertiary Education Scientific Research Project of Guangzhou Municipal Education Bureau(No.202235387)the Guangzhou Science and Technology Project of Guangzhou Municipal Science and Technology Bureau(No.2023A03J0428)the Natural Science Foundation of Guangdong Province,China(No.2024A1515013082)the Guangdong Basic and Applied Basic Research 21 Foundation(No.2021A1515010403).
文摘Background:Alterations in splicing factors contribute to aberrant alternative splicing(AS),which subsequently promotes tumor progression.The splicing factor polypyrimidine tract binding protein 1(PTBP1)has been shown to facilitate cancer progression by modulating oncogenic variants.However,its specific role and underlying mechanisms in hepatocellular carcinoma(HCC)remain to be elucidated.Methods:PTBP1 expression was evaluated in HCC tissues and cell lines.Subsequently,cells were transfected with vectors designed for PTBP1 overexpression or downregulation.The biological function of PTBP1 was assessed in vitro and in vivo using MTS assays,colony formation assays,transwell assays,xenograft formation,tail vein injection,and orthotopic models.Transcriptome analysis was conducted to elucidate the underlying molecular mechanisms.Results:Our findings demonstrated that PTBP1 exhibited elevated expression in HCC cell lines and tissues.Furthermore,its expression positively correlated with overall and disease-free survival rates,as well as tumor grade and stage.PTBP1 knockdown reduced HCC cell proliferation,migration,and invasion in vitro and suppressed hepatocarcinoma xenograft growth and infiltration in vivo.RNA sequencing(RNA-Seq)analysis identified the AS events associated with PTBP1.PTBP1 functionally enhanced cell proliferation,invasion,and migration by modulating the AS of the microtubule-associated protein tau(MAPT)gene and promoting oncogene expression.Notably,the dysregulation of MAPT splicing coincided with increased PTBP1 expression in HCC.Conclusions:PTBP1-guided AS of the MAPT gene enhances tumorigenicity in HCC through activation of the MAPK/ERK pathways.
基金supported by the National Basic Research Program of China (973 Program) (No 2006CB943601)the National Natural Science Foundation of China (NSFC) (No 90919042)
文摘OCT4 is one of the key transcription factors in maintaining the pluripotency and self-renewal of embryonic stem （ES） cells.Human OCT4 can generate two isoforms OCT4A and OCT4B by alternative splicing.OCT4B1 is a recently discovered novel OCT4 spliced variant,which has been considered as a putative marker of stemness.Compared with the OCT4B mRNA,OCT4B1 mRNA is generated by retaining intron 2 as a cryptic exon which contains a TGA stop codon in it.As a result,the protein product of OCT4B1 mRNA could be truncated.Interestingly,we present here that OCT4B1 can indirectly produce the same protein products as OCT4B.We have demonstrated that OCT4B1 mRNA can be spliced into OCT4B mRNA,and encode three protein isoforms.The splicing of OCT4B1 mRNA into OCT4B mRNA can be remarkably inhibited by the mutation of the classical splicing site.Our result suggests that OCT4B mRNA may originate from OCT4B1 mRNA by alternative splicing.
基金financially supported by the National Natural Science Foundation of China (51741908)
文摘Developing effective irrigation and drainage strategies to improve the quality of saline-alkali soil is vital for enhancing agricultural production and increasing economic returns. In this study, we explored how irrigation and drainage modes (flood irrigation, drip irrigation, and sub-surface pipe drainage under drip irrigation) improve the saline-alkali soil in Xinjiang, China. We aimed to study the transport characteristics of soil water and salt under different irrigation and drainage modes, and analyze the effects of the combination of irrigation and drainage on soil salt leaching, as well as its impacts on the growth of oil sunflower. Our results show that sub-surface pipe drainage under drip irrigation significantly reduced the soil salt content and soil water content at the 0–200 cm soil depth. Under sub-surface pipe drainage combined with drip irrigation, the mean soil salt content was reduced to below 10 g/kg after the second irrigation, and the soil salt content decreased as sub-surface pipe distance decreased. The mean soil salt content of flood irrigation exceeded 25 g/kg, and the mean soil desalination efficiency was 3.28%, which was lower than that of drip irrigation. The mean soil desalination rate under drip irrigation and sub-surface pipe drainage under drip irrigation was 19.30% and 58.12%, respectively. After sub-surface drainage regulation under drip irrigation, the germination percentage of oil sunflower seedlings was increased to more than 50%, which further confirmed that combined drip irrigation and sub-surface pipe drainage is very effective in improving the quality of saline-alkali soil and increasing the productivity of agricultural crops.
文摘The removal efficiency of Cu and Zn from swine wastewater was evaluated as effected by three variables: the hydraulic retention time (HRT) (24, 48, 72 and 96 hours), two different plant species (Typha domingensis Pers. and Eleocharis cellulosa) and two different sizes of filter media (5 and 15 mm) using a horizontal sub-surface flow constructed wetland. From the results, a significant difference was observed in the removal efficiency of Cu and Zn with respect to different hydraulic retention times. The best results were obtained in the HRT of 96 hours for Zn where 96% removal of Zn with Typha domingensis Pers. specie with gravel of 15 mm (experimental unit 6) was achieved. For Cu, at 72 hours of HRT, the efficiency was nearly 100% in five of the six study units (1, 2, 3, 5 and 6). In contrast, in experimental unit 4 with gravel of 15 mm and without plants, only 86% Cu removal was achieved.
基金the National Natural Science Foundation of China through Grants 40631006 and 40306005
文摘An Arctic Ocean eddy in sub-surface layer is analyzed in this paper by use of temperature, salinity and current profiles data obtained at an ice camp in the Canada Basin during the second Chinese Arctic Expedition in summer of 2003. In the vertical temperature section, the eddy shows itself as an isolated cold water block at depth of 60 m with a minimum temperature of - 1.5℃, about 0.5℃ colder than the ambient water. Isopycnals in the eddy form a pattern of convex, which indicates the eddy is anticyclonic. Although maximum velocity near 0.4 m s^-1 occurs in the current records observed synchronously, the current pattern is far away from a typical eddy. By further analysis, inertial frequency osci/lations with amplitudes comparable with the eddy velocity are found in the sub-surface layer currents. After filter the inertial current and mean current, an axisymmetric current pattern of an eddy with maximum velocity radius of 5 km is obtained. The analysis of the T-S characteristics of the eddy core water and its ambient waters supports the conclusion that the eddy was formed on the Chukchi Shelf and migrated northeastward into the northern Canada Basin.
文摘The shaping form of an untwisted yarn-end for the air splicer is analyzed and then a cosine curve form which will have practically good splicing effect is proposed. The yarn motion in the splicing nozzle by applying hydrodynamics and the splicing principle of the air splicer are also studied.
文摘The aim of the present study is to improve the capabilities and precision of a recently introduced Sea Surface Acoustic Simulator(SSAS) developed based on optimization of the Helmholtz–Kirchhoff–Fresnel(HKF) method. The improved acoustic simulator, hereby known as the Modified SSAS(MSSAS), is capable of determining sound scattering from the sea surface and includes an extended Hall–Novarini model and optimized HKF method. The extended Hall–Novarini model is used for considering the effects of sub-surface bubbles over a wider range of radii of sub-surface bubbles compared to the previous SSAS version. Furthermore, MSSAS has the capability of making a three-dimensional simulation of scattered sound from the rough bubbly sea surface with less error than that of the Critical Sea Tests(CST) experiments. Also, it presents scattered pressure levels from the rough bubbly sea surface based on various incident angles of sound. Wind speed, frequency, incident angle, and pressure level of the sound source are considered as input data, and scattered pressure levels and scattering coefficients are provided. Finally, different parametric studies were conducted on wind speeds, frequencies, and incident angles to indicate that MSSAS is quite capable of simulating sound scattering from the rough bubbly sea surface, according to the scattering mechanisms determined by Ogden and Erskine. Therefore, it is concluded that MSSAS is valid for both scattering mechanisms and the transition region between them that are defined by Ogden and Erskine.
文摘RNA splicing normally generates stable splice- junction sequences in viruses that are important in the context of virus mimicry. Potential variability in envelop proteins may occur with point-mutations inducing cryptic splice-junctions, which would remain unrecognized by T-memory cells of higher organisms in vaccine trials. Such aberrant splice- junctions result from evolution-specific non-conser- vation of actual splice-junction sites due to mutations;as such, locations of splice-junctions in a test DNA sequence could only be imprecisely specified. Such impreciseness of splice-junction locations (or cryptic sites) in a sequence is evaluated in this study via “noisy” attributes (with associated stochastics) to the mutated subspace;and, relevant fuzzy considerations are invoked with membership attributes expressed in terms of a spatial signal-to-noise ratio (SSNR). That is, SSNR adopted as a membership function expresses the belongingness of a site-region to exon/intron subspaces. An illustrative example with actual (Dengue 1 viral) DNA data is furnished demonstrating the pursuit developed in predicting aberrant splice-junctions at cryptic sites in the test sequence.
文摘This study aims at identifying possible correlations between shapes, types of geophysical anomalies and borehole productivity according to geological and hydrogeotogical contexts. The methodology adopted was a two-pronged one--the first step sought to： （1） interpret the electrical resistivity values from horizontal profiling and vertical electrical sounding implemented in Seno province that preceded the drilling of 513 boreholes; （ii） interpret data from pumping tests carried out on boreholes having a discharge superior to 1 m3/h （＂positive borehole＂） by using Cooper-Jacob＇s method. In the second step, according to geology, authors tried to identify possible correlations between each of the qualitative geophysical parameters： ~shape of anomaly〉〉, tttype of anomaly〉〉 and ~〈type curve〉〉 on the one hand, and hydrogeological parameters such as discharge, alteration thickness, transmissivity and saturated level on the other. The results of this study have shown that the chances of having a positive borehole in Seno province are higher when the type of anomaly is TCC （80%）, shape of anomaly is ＂W＂ and when type curve is ＂H＂ （80%） for all geological formations. Granitic formations are those that record higher discharges while schists record high transmissivity values.
基金This publication was supported by Award Number(BAP,FÖA-2016-6152)from the Research Foundation of the Cukurova University,Adana,Turkey and Principal Investigator,Prof.Dr.Alhan Sariyev“https://bap.cu.edu.tr/.”
文摘Maize being sub-tropical crop is sensitive to water deficit during the early growth stages;particularly clay-rich soil,due to the compaction of the soil.It is well-documented that potential sub-surface drip irrigation(SDI)(Full irrigation;SDIFull(100%field capacity(FC)),Deficit irrigation;SDIDeficit(70%FC))improves water use efficiency,which leads to increased crop productivity;since it has a constraint that SDI excludes soil air around the root-zone during irrigation events,which alter the root function and crop performance.Additionally,in clay-rich soils,the root system of plants generally suffers the limitation of oxygen,particularly the temporal hypoxia,and occasionally from root anoxia;while SDI system accomplishes with the aerating stream of irrigation in the rhizosphere could provide oxygen root environment.The oxygen can be introduced into the irrigation stream of SDI through two ways:the venturi principle,or by using solutions of hydrogen peroxide through the air injection system.Therefore,the application of hydrogen peroxide(H_(2)O_(2);HP)can mitigate the adverse effect of soil compactness and also lead to improving the growth,yield and yield attributes of maize in clay-rich soil.Considering the burning issue,a field study was conducted in consecutive two seasons of 2017 and 2018;where hybrid maize was cultivated as a second crop,to evaluate the effect of liquid-injection of H_(2)O_(2)(HP)into the irrigation stream of SDI on the performance of maize in a clay-rich soil field of Adana,Turkey.When soil water content decreased in 50%of avail-able water,irrigation was performed.The amount of water applied to reach the soil water content to the field capacity is SDIFull(100%FC)and 70%FC of this water is SDIDeficit(70%FC).In the irrigation program,hydrogen peroxide(HP)was applied at intervals of 7 days on average according to available water with and without HP:SDIFull(100%FC)+0 ppm HP with full SDI irrigation;SDIFull(100%FC)+250 ppm HP with deficit SDI irrigation;SDIDeficit(70%FC)+0 ppm HP,SDIDeficit(70%FC)+250 ppm HP and SDIDeficit(70%FC)+500 ppm HP.Deficit irrigation(SDIDeficit(70%FC))program was started from tasseling stage and continued up to the physiological maturity stage with sub-soil drip irrigation.H_(2)O_(2) was applied 3 times during the growing season.Two years’results revealed that the liquid-injection of H_(2)O_(2) into the irrigation stream of SDI improved the growth and yield-related attributes and grain yield of maize.Based on the obtained results,during the extreme climatic condition in the year 2017,SDIFull(100%FC)+250 ppm HP was more effective than SDIFull(100%FC)+0 ppm HP on all traits for relative to full irrigation.While,during the favourable climatic condition in the 2018 season,SDIFull(100%FC)+250 ppm HP was more effective than full irrigation with SDIFull(100%FC)+0 ppm HP for the grain yield,grains,and SPAD value.Accordingly,the most effective treatment was SDIFull(100%FC)+250 ppm HP,as it gave the highest growth and yield-related attributes and grain yield of maize followed by SDIDeficit(70%FC)+250 ppm HP.Therefore,SDIFull with 250 ppm H_(2)O_(2) using as liquid-injection may be recommended to mitigate the adverse effect of soil compactness particularly water-deficit stress in clay-rich soil for the sustainability of maize production.
基金supported by grants from the National Natural Science Foundation of China(Grant Nos.32260085,31860064,31660501,31970609,32260718 and 31901870)the Key Projects of the Applied Basic Research Plan of Yunnan Province(Grant No.202301AS070082)+3 种基金the Start-up fund from Xishuangbanna Tropical Botanical Garden,the‘Top Talents Program in Science and Technology’from Yunnan Province,the Major Science and Technology Project in Yunnan Province(Grant Nos.202102AE090042 and 202202AE090036)the Young and Middle-Aged Academic and Technical Leaders Reserve Talent Program in Yunnan Province(Grant No.202205AC160076)China Postdoctoral Science Foundation(Grant No.2019M653849XB)the High-level Talents Introduction Plan of Yunnan Province-Young Talents Special Project。
文摘The formation of root system architecture(RSA)plays a crucial role in plant growth.OsDRO1 is known to have a function in controlling RSA in rice,however,the role of potato StDRO2,a homolog of rice OsDRO1,in root growth remains unclear.In this study,we obtained potato dro2 mutant lines by Clustered Regularly Interspaced Short Palindromic Repeats-CRISPR-Associated 9(CRISPR/Cas9)-mediated genome editing system.The mutant lines were generated from a splicing defect of the StDRO2 intron 1,which causes a nonsense mutation in StDRO2.Furthermore,the secondary structure of StDRO2 mRNA analyzed with RNAfold Web Server was altered in the dro2 mutant.Mutation of StDRO2 conveys potato adaptation through changing the RSA via alteration of auxin transport under drought stress.The potato dro2 lines showed higher plant height,longer root length,smaller root growth angle and increased tuber weight than the wild-type.The alteration of RSA was associated with a disturbance of IAA distribution in the dro2 mutant,and the levels of StPIN7 and StPIN10 detected by using real-time PCR were up-regulated in the roots of potato dro2 lines grown under drought stress.Moreover,the microRNAs(miRNAs)PmiREN024536 and PmiREN024486 targeted the StDRO2 gene,and auxin positively and negatively regulated the expression of StDRO2 and the miRNAs PmiREN024536 and PmiREN024486,respectively,in the potato roots.Our data shows that a regulatory network involving auxin,StDRO2,PmiREN024536 and PmiREN024486 can control RSA to convey potato fitness under drought stress.
基金supported by the National Natural Science Foundation of China(32072126 and 32230075)the Shandong Provincial Natural Science Foundation(ZR2019MC005).
文摘Pentatricopeptide repeat(PPR)proteins are a large group of eukaryote-specific RNA-binding proteins that play pivotal roles in plant organelle gene expression.Here,we report the function of PPR21 in mitochondrial intron splicing and its role in maize kernel development.PPR21 is a typical P-type PPR protein targeted to mitochondria.The ppr21 mutants are arrested in embryogenesis and endosperm development,leading to embryo lethality.Null mutations of PPR21 reduce the splicing efficiency of nad2 intron 1,2,and 4 and impair the assembly and activity of mitochondrial complex I.Previous studies show that the P-type PPR protein EMP12 is required for the splicing of identical introns.However,our protein interaction analyses reveal that PPR21 does not interact with EMP12.Instead,both PPR21 and EMP12 interact with the small MutS-related(SMR)domain-containing PPR protein 1(PPR-SMR1)and the short P-type PPR protein 2(SPR2).PPR-SMR1 interacts with SPR2,and both proteins are required for the splicing of many introns in mitochondria,including nad2 intron 1,2,and 4.These results suggest that a PPR21-(PPR-SMR1/SPR2)-EMP12 complex is involved in the splicing of nad2 introns in maize mitochondria.
文摘In this study,we examine the problem of sliced inverse regression(SIR),a widely used method for sufficient dimension reduction(SDR).It was designed to find reduced-dimensional versions of multivariate predictors by replacing them with a minimally adequate collection of their linear combinations without loss of information.Recently,regularization methods have been proposed in SIR to incorporate a sparse structure of predictors for better interpretability.However,existing methods consider convex relaxation to bypass the sparsity constraint,which may not lead to the best subset,and particularly tends to include irrelevant variables when predictors are correlated.In this study,we approach sparse SIR as a nonconvex optimization problem and directly tackle the sparsity constraint by establishing the optimal conditions and iteratively solving them by means of the splicing technique.Without employing convex relaxation on the sparsity constraint and the orthogonal constraint,our algorithm exhibits superior empirical merits,as evidenced by extensive numerical studies.Computationally,our algorithm is much faster than the relaxed approach for the natural sparse SIR estimator.Statistically,our algorithm surpasses existing methods in terms of accuracy for central subspace estimation and best subset selection and sustains high performance even with correlated predictors.
文摘目的探讨虚拟仿真结合改良自我导向、问题导向、终身学习、综合临床案例练习(selfdirected,problem-oriented,lifelong learning,integrated clinical case exercise,SPLICE)教学法对临床医学专硕生批判性思维能力的影响。方法选取2024年1—6月在锦州医科大学附属第一医院皮肤科进行住院医师规范化培训(简称“住培”)的64名2023级临床医学专硕生为研究对象。采用随机分组法将其分为试验组(n=32)和对照组(n=32)。试验组采用虚拟仿真结合改良SPLICE教学,对照组采用传统教学方式。比较两组学生批判性思维能力、学业自我效能感、理论考核和临床操作技能考核成绩。结果试验组医学专硕生的批判性思维能力、学业自我效能感、理论考核和临床操作技能考核成绩均高于对照组,差异具有统计学意义(均P<0.001)。结论虚拟仿真结合改良SPLICE教学法有助于提升医学专硕生的批判性思维能力与学业自我效能感,有利于其更好地掌握专业理论和临床操作技能,值得在临床教学中进一步推广应用。
基金supported in part by the Natural Science Foundation of Shandong Province(no.ZR2022QH373,ZR2022QH292 and ZR2023MH2474).
文摘ObjectivesThe PTPRQ gene is essential for preserving the structure and function of stereocilia in inner ear.However,research on splicing mutations within this gene is limited.This study aims to investigate novel splicing mutations in PTPRQ,clarify their molecular mechanisms,and provide new insights into the genetic factors associated with hearing loss,ultimately enhancing diagnostic accuracy.MethodClinical data and peripheral blood samples were obtained from members of a family with congenital hearing loss.Variants were identified through high-throughput sequencing and confirmed by Sanger sequencing to ensure genealogical co-segregation.The splicing effects of PTPRQ variants were evaluated using bioinformatics tools and minigene assays.ResultsWe used whole exome sequencing to identify novel double compound heterozygous splice-altering variants(c.5426+1 G>A and c.6603-3 T>G)in the PTPRQ gene with DFNB84A.We molecularly characterized these variants,and they were found to co-segregate with the disease within the family.Minigene assays and Sanger sequencing confirmed that the c.6603-3 T>G variant caused exon 43 skipping,resulting in a frameshift mutation(p.Ser2201ArgfsTer112).Further bioinformatic analysis supported these findings.ConclusionsThis study identifies a novel compound heterozygous splicing variant in the PTPRQ gene in a Chinese family with DFNB84A,expanding the known spectrum of PTPRQ mutations.These findings enhance the understanding of PTPRQ-related hearing loss and may aid in early diagnosis,prevention,and therapeutic strategies.
基金funded by the National Natural Science Foundation of China(32202393)the Natural Science Foundation of Shandong Province,China(ZR2021QC190)+1 种基金the Science and Technology Benefiting the People Demonstration Project of Qingdao,China(24-1-8-xdny-10-nsh)the Qingdao Agricultural University High-level Talent Fund,China(663/1120101)。
文摘Caspases,which play key roles in cell apoptosis,undergo alternative splicing to form different splicing variants that can regulate the apoptotic process.Lepidopteran insect caspases undergo alternative splicing,although the functions of their splicing variants are still unclear.The Spodoptera exigua caspase-5(SeCaspase-5)gene was cloned and found to produce four different splicing variants with different gene sequences and protein functional domains,which were named SeCaspase-5a,SeCaspase-5b,SeCaspase-5c and SeCaspase-5d.Overexpression of these variants in S.exigua cells(Se-3)showed that SeCaspase-5a had a proapoptotic function,whereas SeCaspase-5b,SeCaspase-5c and SeCaspase-5d did not.Semi-qPCR analysis revealed that the expression of the SeCaspase-5 variants significantly differed during Autographa californica multiple nucleopolyhedrovirus(AcMNPV)infection.Furthermore,the SeCaspase-5 variants were constructed into the AcMNPV bacmid and transfected into Se-3 cells,which revealed that SeCaspase-5a promoted cell apoptosis and reduced virus production,whereas SeCaspase-5b,SeCaspase-5c and SeCaspase-5d did not promote cell apoptosis but instead increased virus production.Moreover,an analysis of the interactions between the SeCaspase-5 variants revealed that SeCaspase-5a directly interacted with SeCaspase-5b,SeCaspase-5c and SeCaspase-5d.Coexpression of these variants in Se-3 cells also revealed that SeCaspase-5b,SeCaspase-5c and SeCaspase-5d inhibited the proapoptotic function of SeCaspase-5a,resulting in a reduction in the percentage of apoptotic cells by about 20%.These results indicate that SeCaspase-5 undergoes alternative splicing and is involved in regulating the apoptosis induced by baculovirus infection.These findings increase our understanding of the functions of lepidopteran insect caspases and provide new insights into the mechanism of host-cell apoptosis induced by baculoviruses.
基金supported by the National Key Research and Development Program of China(2021YFF1000301)the National Natural Science Foundation of China(31771805)。
文摘Heat stress is a major threat to maize(Zea mays L.)production worldwide.Heat shock transcription factors(HSFs)play vital roles in plant responses to heat stress.However,the molecular and genetic mechanisms underlying HSF-meditated thermotolerance in maize remain largely unexplored.In this study,we demonstrate that the alternative splicing of ZmHsf23 modulates heat stress tolerance in maize.Hsf23 produced two functional transcripts,Hsf23L and Hsf23S,which differ by the presence of a cryptic mini-exon in Hsf23L that is spliced out in Hsf23S.Both transcripts were strongly induced by heat stress.Mutants lacking Hsf23L alone(hsf23l)or both Hsf23L and Hsf23S(hsf23l23s)exhibited increased susceptibility to heat stress,whereas overexpression of Hsf23S enhanced heat stress tolerance in maize.Subsequently,we found that Hsf23S positively regulates heat stress tolerance by directly activating the transcription of three sHSP genes(Hsp16.9,Hsp17.2,and Hsp18a)and TIL1 gene.In addition,Hsf23L physically interacted with Hsf23S and enhanced the transcriptional activation of Hsf23S on the sHSPs and TIL1 promoters.Notably,genetic analysis suggested that co-overexpression of Hsf23L and Hsf23S further improves heat tolerance of the transgenic plants.Taken together,these results reveal two splicing variants of ZmHsf23 cooperatively regulate maize heat tolerance,thus highlighting potential value of ZmHsf23 in breeding heat-tolerant maize varieties.
基金funded by Guizhou Province Science and Technology Plan Project Qiankehe Foundation-ZK[2023]General 360,362Science and Technology Fund project of Guizhou Provincial Health Commission(gzwkj-2022-09,gzwkj-2023-035)+1 种基金National Natural Science Foundation Cultivation Project of Guizhou Medical University(21NSFCP14,gyfynsfc-2022-25)The PhD Scientific Research Launch Fund Project of the Affiliated Hospital of Guizhou Medical University(gyfybsky-2022-02).
文摘Background:Glioblastoma(GBM)is one of the most malignant types of central nervous system tumors.Oxygen deprivation in the tumor microenvironment is thought to be an important factor in promoting GBM progression.However,the mechanisms of hypoxia-promoted tumor progression remain elusive.Methods:Alternative splicing of diacylglycerol kinase gamma(DGKG)-Δexon13 was amplified and verified by PCR-Sanger sequencing.The functions of DGKG and DGKG-Δexon13 were analyzed by Cell counting kit-8(CCK-8),Transwell,Matrigeltranswell experiments,and in vivo orthotropic GBM animal models.Transcriptome analyses were done to find out the regulated genes.Results:In this study,we found that a new transcript DGKG-Δexon13 was generated in GBM under hypoxia via alternative splicing.Moreover,the results of CCK-8,Transwell,and Matrigel-transwell experiments showed that the proliferation,migration,and invasion abilities of U87-MG and T98G were decreased after DGKG knockdown.Compared to wild-type DGKG,DGKG-Δexon13 overexpression significantly promoted cellular proliferation,migration,and invasion abilities in GBM.Furthermore,in vivo,orthotropic GBM animal models analysis showed that the tumor volumes were much smaller in the DGKG knockdown group.However,the tumor sizes in the DGKG and DGKG-Δexon13 rescue groups were restored,especially in the DGKG-Δexon13 group.Transcriptome analysis revealed that MORC1,KLHDC7B,ATP1A2,INHBE,TMEM119,and FGD3 were altered significantly when DGKG was knocked down.IL-16,CCN2,and EFNB3 were specifically regulated by DGKG-Δexon13.Conclusions:Our study found that hypoxia-induced alternative splicing transcript DGKG-Δexon13 promotes GBM proliferation and infiltration,which might provide a new potential target for the clinical treatment and diagnosis of GBM.
基金supported by the Major Program of National Agricultural Science and Technology of China(NK20220607)the Sichuan Science and Technology Program,China(2023YFH0041)。
文摘Starch biosynthesis is a complex process that relies on the coordinated action of multiple enzymes.Resistant starch is not digested in the small intestine,thus preventing a rapid rise in the glycemic index.Starch synthase 2a(SS2a)is a key enzyme in amylopectin biosynthesis that has significant effects on starch structure and properties.In this study,we identified an ss2a null mutant(M3-1413)with a single base mutation from an ethyl methane sulfonate(EMS)-mutagenized population of barley.The mutation was located at the 3'end of the first intron of the RNA splicing receptor(AG)site,and resulted in abnormal RNA splicing and two abnormal transcripts of ss2a,which caused the inactivation of the SS2a gene.The starch structure and properties were significantly altered in the mutant,with M3-1413 containing lower total starch and higher amylose and resistant starch levels.This study sheds light on the effect of barley ss2a null mutations on starch properties and will help to guide new applications of barley starch in the development of nutritious food products.
