Genetic analysis and candidate gene identification of salt tolerancerelated traits in maize

Soil salinization poses a threat to maize production worldwide,but the genetic mechanism of salt tolerance in maize is not well understood.Therefore,identifying the genetic components underlying salt tolerance in maize is of great importance.In the current study,a teosinte-maize BC2F7 population was used to investigate the genetic basis of 21 salt tolerance-related traits.In total,125 QTLs were detected using a high-density genetic bin map,with one to five QTLs explaining 6.05–32.02%of the phenotypic variation for each trait.The total phenotypic variation explained(PVE)by all detected QTLs ranged from 6.84 to 63.88%for each trait.Of all 125 QTLs,only three were major QTLs distributed in two genomic regions on chromosome 6,which were involved in three salt tolerance-related traits.In addition,10 pairs of epistatic QTLs with additive effects were detected for eight traits,explaining 0.9 to 4.44%of the phenotypic variation.Furthermore,18 QTL hotspots affecting 3–7 traits were identified.In one hotspot(L5),a gene cluster consisting of four genes(ZmNSA1,SAG6,ZmCLCg,and ZmHKT1;2)was found,suggesting the involvement of multiple pleiotropic genes.Finally,two important candidate genes,Zm00001d002090 and Zm00001d002391,were found to be associated with salt tolerance-related traits by a combination of linkage and marker-trait association analyses.Zm00001d002090 encodes a calcium-dependent lipid-binding(CaLB domain)family protein,which may function as a Ca^(2+)sensor for transmitting the salt stress signal downstream,while Zm00001d002391 encodes a ubiquitin-specific protease belonging to the C19-related subfamily.Our findings provide valuable insights into the genetic basis of salt tolerance-related traits in maize and a theoretical foundation for breeders to develop enhanced salt-tolerant maize varieties.

Fine-mapping and primary analysis of candidate genes associated with seed coat color in mung bean(Vigna radiata L.)

Seed coat color affects the appearance and commodity quality of mung beans(Vigna radiata L.).The substances that affect mung bean seed coat color are mainly flavonoids,which have important medicinal value.Mapping the seed coat color gene in mung beans would facilitate the development of new varieties and improve their value.In this study,an F2 mapping population consisting of 546 plants was constructed using Jilv9(black seed coat)and BIS9805(green seed coat).Using bulk segregated analysis(BSA)sequencing and kompetitive allele-specific PCR(KASP)markers,the candidate region related to seed coat color was finally narrowed to 0.66 Mb on chromosome(Chr.)4 and included eight candidate genes.Combined transcriptome and metabolome analyses showed that three of the eight candidate genes(LOC106758748,LOC106758747,and LOC106759075)were differentially expressed,which may have caused the differences in flavonoid metabolite content between Jilv9 and BIS9805.These findings can provide a research basis for cloning the genes related to seed coat color and accelerate molecular markerassisted selection breeding in mung beans.

Comparative analysis of dideoxy sequencing,the KRAS StripAssay and pyrosequencing for detection of KRAS mutation

AIM:To compare the differences between dideoxy sequencing/KRAS StripAssay/pyrosequencing for detection of KRAS mutation in Chinese colorectal cancer (CRC) patients.METHODS:Formalin-f ixed, paraff in-embedded (FFPE) samples with tumor cells ≥ 50% were collected from 100 Chinese CRC patients at Beijing Cancer Hospital. After the extraction of genome DNA from FFPE samples, fragments contained codons 12 and 13 of KRAS exon 2 were amplified by polymerase chain reaction and analyzed by dideoxy sequencing, the KRAS Strip Assay and pyrosequencing. In addition, the sensitivities of the 3 methods were compared on serial dilutions (contents of mutant DNA: 100%,50%,20%, 5%,10%, 5%,1%,0%) of A549 cell line DNA (carrying the codon 12 Gly>Ser mutation) into wild-type DNA (human normal intestinal mucosa). The results of dideoxy sequencing,the KRAS StripAssay and pyrosequencing were analyzed by Chromas Software, Collector forKRAS Strip Assay and the pyrosequencing PyroMarkTM Q24 system, respectively.RESULTS: Among 100 patients, KRAS mutations were identif ied in 34%, 37% and 37% of patients by dideoxy sequencing, the KRAS StripAssay and pyrosequencing, respectively. The sensitivity was highest with the KRAS Strip Assay (1%), followed by pyrosequencing (5%), and dideoxy sequencing was lowest (15%). Six different mutation types were found in this study with 3 main mutations Gly12 Asp (GGT>GAT), Gly12 Val (GGT>GTT) and Gly13 Asp (GGC>GAC). Thirty-three patients were identifi ed to have KRAS mutations by the 3 methods, and a total of 8 patients had conflicting results between 3 methods: 4 mutations not detected by dideoxy sequencing and the KRAS StripAssay were identified by pyrosequencing; 3 mutations not detected by dideoxy sequencing and pyrosequencing were identif ied by the KRAS StripAssay; and 1 mutation not detected by pyrosequencing was conf irmed by dideoxy sequencing and the KRAS StripAssay. Among these discordant results, the results identif ied by dideoxy sequencing were consistent either with the KRAS StripAssay or with pyrosequencing, which indicated that the accuracy of dideoxy sequencing was high. CONCLUSION: Taking a worldwide view of reports and our results,dideoxy sequencing remains the most popular method because of its low cost and high accuracy.

Molecular mapping and candidate gene analysis of the semi-dominant gene Vestigial glume1 in maize

The glume is an organ of the maize spikelet and plays important roles in anther and kernel development.Vestigial glume1(Vg1)is a classic mutant associated with ligule and glume development.Here we report the phenotypic characterization,fine mapping,and candidate gene analysis of the Vg1 mutant.Vg1 is a semi-dominant and pleiotropic gene,and also affects plant height,ear height,and tassel length.Vg1 ligule degeneration begins at the first leaf,and the Vg1 tassel and ear can be distinguished from those of wild-type plants when their lengths reach respectively 55 mm and 51 mm.Using a BC3 mapping population of 11,445 plants,we delimited the Vg1 functional site to an interval of 7.4 kb,flanked by the markers InDelLM and CRM6.A putative cyclopropane fatty-acid synthase gene(ZmCPA-FAS1)was hypothesized to underlie the mutant phenotype.We detected a Helitron insertion in the sixth intron of ZmCPAFAS1.Its presence caused abnormal alternative splicing of ZmCPA-FAS1 that conferred new characteristics on the Vg1 mutant.These findings are a basis for further discovery of the molecularmechanism underlying glume development and a potential guide formaize breeding of small-glume varieties,especially sweet corn breeding.

Meta-analysis of soybean amino acid QTLs and candidate gene mining

The composition and quantity of amino acids influence the protein content and nutritional value of soybeans and also have an important impact upon soybean quality. After integrating and proofreading 140 original QTLs associated with amino acid contentfrom soybase（http：//www.soybase.org/）, 138 QTLs were further analyzed to determine high-confidence QTL regions. Meta-analysis was first carried out using the Bio Mercator ver. 2.1 software, yielding 33 consensus QTLs. The consensus QTL confidence intervals（CIs） ranged from 0.07 to 19.85 Mb. Next, the overview method was used to optimize the CIs, and 57 ＂real＂ QTLs were mapped. Candidate genes in the consensus QTL regions were obtained from Phytozome and were annotated using the Gene Ontology（GO）, Kyoto Encyclopedia of Genes and Genomes（KEGG）, Swissprot, and gene annotation databases. Finally, 16 unpublished candidate genes controlling the content of five types of amino acids were identified with Blast. These results laid the foundation for fine mapping of soybean amino acid-related QTLs and marker-assisted selection.

Identification of candidate biomarkers correlated with pathogenesis of postoperative peritoneal adhesion by using microarray analysis

BACKGROUND Postoperative peritoneal adhesion(PPA),characterized by abdominal pain,female infertility,and even bowel obstruction after surgery,has always been a major concern.The occurrence and formation of adhesion are from complex biological processes.However,the molecular mechanisms underlying the basis of microarray data profile,followed by peritoneal adhesion formation,are largely unknown.AIM To reveal the underlying pathogenesis of PPA at the molecular level.METHODS The gene expression profile was retrieved from the Gene Expression Omnibus database for our analysis.We identified a panel of key genes and related pathways involved in adhesion formation using bioinformatics analysis methods.We performed quantitative PCR and western blotting in vivo to validate the results preliminarily.RESULTS In total,446 expressed genes were altered in peritoneal adhesion.We found that several hub genes(e.g.,tumor necrosis factor,interleukin 1 beta,interleukin 6,CX-C motif chemokine ligand 1,C-X-C motif chemokine ligand 2)were marked as significant biomarkers.Functional analysis suggested that these genes were enriched in the Toll-like receptor signaling pathway.According to the Kyoto Encyclopedia of Genes and Genomes pathway and published studies,TLR4,myeloid differentiation primary response protein 88(MyD88),and nuclear factor kappa B(NF-κB)played essential roles in Toll-like signaling transduction.Here,we obtained a regulatory evidence chain of TLR4/MyD88/NF-κB/inflammatory cytokines/peritoneal adhesion involved in the pathogenesis of postoperative adhesion.The results of the microarray analysis were verified by the animal experiments.These findings may extend our understanding of the molecular mechanisms of PPA.CONCLUSION The regulatory evidence chain of TLR4/MyD88/NF-κB/inflammatory cytokines/peritoneal adhesion may play key roles in the pathogenesis of PPA.Future studies are required to validate our findings.

Characterization and Candidate Gene Analysis of the Yellow-Green Leaf Mutant ygl16 in Rice(Oryza sativa L.)

Leaf color mutants are ideal materials for studying many plant physiological and metabolic processes such as photosynthesis,photomorphogenesis,hormone physiology and disease resistance.In this study,the genetically stable yellow-green leaf mutant ygl16 was identified from mutated“Xinong 1B”.Compared with the wild type,the pigment concentration and photosynthetic capacity of the ygl16 decreased significantly.The ultrastructural observation showed that the distribution of thylakoid lamellae was irregular in ygl16 chloroplasts,and the grana and matrix lamellae were blurred and loose in varied degrees,and the chloroplast structure was disordered,while the osmiophilic corpuscles increased.The results of the genetic analysis and mapping showed that the phenotype of ygl16 was controlled by a pair of recessive nuclear gene.The gene located in the 56Kb interval between RM25654 and R3 on the long arm of chromosome 10.The sequencing results showed that the 121st base of the first intron of the candidate gene OsPORB/FGL changed from A to T in the interval.qRT-PCR results showed that the expression of chlorophyll synthase-related genes in the mutant decreased.

Regionalization of epididymal duct and epithelium in rats and mice by automatic computer-aided morphometric analysis

Aim： To establish a rat and mouse epididymal map based on the use of the Epiquatre automatic software for histologic image analysis. Methods： Epididymides from five adult rats and five adult mice were fixed in alcoholic Bouin＇s fixative and embedded in paraffin. Serial longitudinal sections through the medial aspect of the organ were cut at 10 jam and stained with hematoxylin and eosin. As determined from major connective tissue septa, nine subdivisions of the rat epididymis and seven for the mouse were determined, consisting of five sub-regions in the caput （rat and mouse）, one （mouse） or three （rat） in the corpus and one in the cauda （rat and mouse）. Using the Epiquatre software, several tubular, luminal and epithelial morphometric parameters were evaluated. Results： Statistical comparison of the quantitative parameters revealed regional differences （2-5 in the rat, 3-6 in the mouse, dependent on parameters） with caput regions 1 and 2 being largely distinguishable from the similar remaining caput and corpus, which were in turn recognizable from the cauda regions in both species. Conclusion： The use of the Epiquatre software allowed us to establish regression curves for different morphometric parameters that can permit the detection of changes in their values under different pathological or experimental conditions. （Asian J Androl 2005 Sep; 7： 267-275）

Bioinformatics Analysis Raises Candidate Genes in Blood for Early Screening of Parkinson's Disease

Parkinson＇s disease （PD） is a typical degenerative disease, which is characterized by the most obvious symptoms of movement dysfunction, including shaking, rigidity, slowness of movement and difficulty in walking and gait. This disease can not be clearly identified through laboratory tests at present, thus application of high-throughput technique in studying the expression profiles of PD helps to find the genetic markers for its early diagnosis. Studies on expression profiles of neurodegenerative diseases have revealed the novel genes and pathways involved in the progress of illness. In this study, the expression profiles of PD in blood were compared, showing that 181 differentially expressed genes （DEG） exhibit a similar expression trend both in patients and in normal controls.

Business Analysis of Didi Chuxing in the Chinese Ride-hailing Market

This essay presents my analysis of Didi Chuxing’s ride-hailing business in the Chinese market considering economic concepts and tools such as market structure,economies of scale and price anchoring to understand aspects of Didi Chuxing such as its cost structure,pricing strategies and marketing methods.As the largest ride-hailing platform in China,the success of Didi Chuxing can offer important guidelines for the healthy development and expansion of this industry to other countries.

Bioinformatic analysis reveals novel biomarkers and candidate drug compositions in prostate cancer

As prostate cancer(PC)patients do more and more genome sequencing,we can predict prognosis through individual oncogenic mutations.Although great success have been made to clarify the incidence of PC,the mechanisms was not completely understood.Recurrence and metastasis of PC remains to be resolved,and novel therapeutic targets need to be found urgently.Microarray datasets GSE6919,GSE55945 and GSE46602 about the PC tissues vs.normal organizations,were obtained from Gene Expression Omnibus.In this study,86 differentially expressed genes were determined having more important clinical significance in the process of PC.29 hub genes significantly enriched in biological processes were analyzed using Cytoscape.The function of these hub genes included the effect of cellular process,skeletal system development,cholesterol transport,regulation of protein oligomerization and cellular component biogenesis,enzyme inhibitor activity and so on.The three of these hub genes were picked out because of their relationships,which can be used as a potential target for the diagnosis and the direction of therapy.And drug predictions were designed for these candidate target molecules,providing direction for future treatment of PC.

Analysis and Strategy Research on the Mathematics Learning Situation of the Postgraduate Candidates

This study takes the students who intend to study for graduate students as the research object,and makes a questionnaire survey on the mathematics learning situation and learning platform of the postgraduate candidates.Questionnaires were distributed online and offline,and a total of 326 valid questionnaires were collected.Based on 326 questionnaires,this paper analyzes the current learning situation of mathematics in the postgraduate entrance examination and studies the strategies based on the present situation,which points out the direction for the construction of mathematics learning and communication platform based on the needs of postgraduate entrance examination.

Integrated analysis of transcriptomic and proteomic data from tree peony(P.ostii)seeds reveals key developmental stages and candidate genes related to oil biosynthesis and fatty acid metabolism

Tree peony(Paeonia section Moutan DC.)seeds are an excellent source of beneficial natural compounds that promote health,and they contain high levels of alpha-linolenic acid(ALA).In recent years,tree peony has been emerging as an oil crop.Therefore,combined analysis of the transcriptome and proteome of tree peony(P.ostii)seeds at 25,32,39,53,67,81,88,95,and 109 days after pollination(DAP)was conducted to better understand the transcriptional and translational regulation of seed development and oil biosynthesis.A total of 38,482 unigenes and 2841 proteins were identified.A total of 26,912 differentially expressed genes(DEGs)and 592 differentially expressed proteins(DEPs)were clustered into three groups corresponding to the rapid growth,seed inclusion enrichment and conversion,and late dehydration and mature stages of seed development.Fifteen lipid metabolism pathways were identified at both the transcriptome and proteome levels.Pathway enrichment analysis revealed that a period of rapid fatty acid biosynthesis occurred at 53–88 DAP.Furthermore,211 genes and 35 proteins associated with the fatty acid metabolism pathway,63 genes and 11 proteins associated with the biosynthesis of unsaturated fatty acids(UFAs),and 115 genes and 24 proteins associated with ALA metabolism were identified.Phylogenetic analysis revealed that 16 putative fatty acid desaturase(FAD)-encoding genes clustered into four FAD groups,eight of which exhibited the highest expression at 53 DAP,suggesting that they play an important role in ALA accumulation.RT-qPCR analysis indicated that the temporal expression patterns of oil biosynthesis genes were largely similar to the RNA-seq results.The expression patterns of fatty acid metabolism-and seed development-related proteins determined by MRM were also highly consistent with the results obtained in the proteomic analysis.Correlation analysis indicated significant differences in the number and abundance of DEGs and DEPs but a high level of consistency in expression patterns and metabolic pathways.The results of the present study represent the first combined transcriptomic and proteomic analysis of tree peony seeds and provide insight into tree peony seed development and oil accumulation.

RNA-seq analysis of Paris polyphylla var.yunnanensis roots identified candidate genes for saponin synthesis

Paris polyphylla Smith var.yunnanensis(Franch.) Hand.-Mazz.is a rhizomatous,herbaceous,perennial plant that has been used for more than a thousand years in traditional Chinese medicine.It is facing extinction due to overharvesting.Steroids are the major therapeutic components in Paris roots,the commercial value of which increases with age.To date,no genomic data on the species have been available.In this study,transcriptome analysis of an 8-year-old root and a 4-year-old root provided insight into the metabolic pathways that generate the steroids.Using Illumina sequencing technology,we generated a high-quality sequence and demonstrated de novo assembly and annotation of genes in the absence of prior genome information.Approximately 87,577 unique sequences,with an average length of 614 bases,were obtained from the root cells.Using bioinformatics methods,we annotated approximately 65.51% of the unique sequences by conducting a similarity search with known genes in the National Center for Biotechnology Information's non-redundant database.The unique transcripts were functionally classified using the Gene Ontology hierarchy and the Kyoto Encyclopedia of Genes and Genomes database.Of 3082 genes that were identified as significantly differentially expressed between roots of different ages,1518(49.25%) were upregulated and 1564(50.75%) were downregulated in the older root.Metabolic pathway analysis predicted that 25 unigenes were responsible for the biosynthesis of the saponins steroids.These data represent a valuable resource for future genomic studies on this endangered species and will be valuable for efforts to genetically engineer P.polyphylla and facilitate saponin-rich plant development.

Combined QTL mapping,GWAS and transcriptomic analysis revealed a candidate gene associated with the timing of spring bud flush in tea plant(Camellia sinensis)

Dear Editor,The timing of the spring bud f lush(TBF)is a crucial agro-nomic trait for the tea plant,as it strongly influences the yield and economic value of harvested fresh tea leaves.The TBF of tea plant is generally defined as the date when>30%of the growing tender shoots have reached the stage of one bud with one to three leaves,referred as to the stage of one and a bud,two and a bud,and three and a bud,respectively.

Aberrant expression of BDNF might serve as a candidate target for cocaine- induced psychosis: insights from bioinformatics analysis and microarray validation

Background Cocaine use disorder(CUD)and associated psychosis are major public health issues worldwide,along with high relapse outcome and limited treatment options.Exploring the molecular mechanisms underlying cocaine-induced psychosis(CIP)could supply integrated insights for understanding the pathogenic mechanism and potential novel therapeutic targets.Aims The aim of the study was to explore common alterations of CUD-schizophrenia target genes and identify core risk genes contributing to CIP through data mining and network pharmacology approach.Methods Target genes of CUD were obtained from GeneCards,Comparative Toxicogenomics Database,Swiss Target Prediction platform and PubChem.Schizophrenia-related target genes were derived from DisGeNET,GeneCards,MalaCards and Online Mendelian Inheritance in Man databases.Then,the overlap genes of these two sets were regarded as risk genes contributing to CIP.Based on these CUD-schizophrenia target genes,functional annotation and pathway analysis were performed using the clusterProfiler package in R.Protein-protein interaction network construction and module detection were performed based on the Search Tool for the Retrieval of Interacting Genes(STRING)database and Cytoscape software.Gene expression datasets GSE54839 and GSE93577 were applied for data validation and diagnostic capacity evaluation of interested hub genes.Results A total of 165 CUD-schizophrenia target genes were obtained.These genes were mainly contributing to chemical synaptic transmission,neuropeptide hormone activity,postsynaptic membrane and neuroactive ligand-receptor interaction pathway.Network analysis and validation analysis indicated that BDNF might serve as an important risk gene in mediating CIP.Conclusions This study generates a holistic view of CIP and provides a basis for the identification of potential CUD-schizophrenia target genes involved in the development of CIP.The abnormal expression of BDNF would be a candidate therapeutic target underlying the pathogenesis of CUD and associated CIP.

Fine mapping and candidate gene analysis of qGL10 affecting rice grain length

Grain size influences the yield and quality of rice(Oryza sativa L.),and grain length is one of the component traits of grain size.In this study,a near-isogenic line LB3 with long grain size was constructed using japonica rice cultivar 02428,with short grain size,as the recipient parent and indica rice cultivar ZYX,with long grain size,as the donor parent,by multi-generation backcrossing and selfing.BSA-seq was used for preliminary QTL mapping and InDel markers were developed to fine map the locus.The major QTL,tentatively named qGL10,for grain length was located in a 128.45 kb region of chromosome 10.Combined with haplotype analysis of rice varieties,expression pattern analysis of candidate genes suggested LOC_Os10g39130(OsMADS56)as a candidate gene.Sequence alignment of OsMADS56 in 02428 and LB3 revealed that there were 15 SNPs in the promoter region and four in the coding region.Further haplotype analysis suggested that SNP9(G/A)located in the TGTCACA motif might account for the different expression levels of OsMADS56 in 02428 and LB3.These results lay a foundation for the application of qGL10 in molecular breeding of new rice varieties.

Comparative transcriptome analysis on candidate genes involved in lipid biosynthesis of developing kernels for three walnut cultivars in Xinjiang

Walnut(Juglans regia L.)is a good source of lipids and polyunsaturated fatty acids(PUFAs).In order to explore the biosynthesis molecular mechanism of oil accumulation and fatty acid(FA)synthesis in walnut,the samples at different development periods of three walnut cultivars,’Zhipi’(ZP),’Xinwu 417’(W417)and’Xinwen 81’(W81)were collected for transcriptomic analysis.The analysis of oil accumulation and FA profiles showed that the oil content in mature walnut kernel was nearly 70%,and over 90%of FAs were PUFAs.We identified 126 candidate genes including 64 genes for FA de novo synthesis,45 genes for triacylglycerol assembly,and 17 genes for oil bodies involved in lipid biosynthesis by RNA-sequencing.Ten key enzymes including ACCase,LACS6,LACS8,SAD,FAD2,FAD3,LPAAT1,DGAT2,PDAT2,and PLC encoded by 19 genes were highly associated with lipid biosynthesis.Quantitative PCR analysis further validated 9 important genes,and the results were well consistent with our transcriptomic data.Finally,5 important transcription factors including WRI1,ABI3,FUS3,PKL and VAL1 were identified,and their main regulatory genes might contain ACCase,KASII,LACS,FAD3 and LPAAT which were determined through correlation analysis of expression levels for 27 walnut samples.These findings will provide a comprehensive understanding and valuable information on the genetic engineering and molecular breeding in walnut.

Did the Revision of the Japanese Medical Payment System Work Properly?—An Analysis of Averages and Variances of Length of Hospital Stay for Type 2 Diabetes Patients by Individual Hospital

We evaluated the effects of the 2010 revision of the medical payment system on the length of stay (LOS). In this analysis, we assessed not only the average length of stay (ALOS), but also variance of LOS at individual hospitals. We used a dataset of 18,641 type 2 diabetes patients collected from 51 general hospitals. The variables found to affect LOS were age, comorbidities, complications, acute hospitalization, introduced by other hospitals, winter, one-week hospitalization, specific hospitalization period, and principal diseases coded E11.5, E11.6 and E11.7. Although the effect was marginal, the 2010 revision did reduce ALOS, and the reduction was larger as ALOS became longer. On the other hand, we did not find that the variance of LOS within hospitals became smaller. The results of the study suggest that new incentives and assistance to hospitals to help them make efficient use of medical information are needed.

Identification of potential candidate proteins for reprogramming spinal cord-derived astrocytes into neurons:a proteomic analysis

Our previous study has confirmed that astrocytes overexpressing neurogenic differentiation factor 1(NEUROD1)in the spinal cord can be reprogrammed into neurons under in vivo conditions.However,whether they can also be reprogrammed into neurons under in vitro conditions remains unclear,and the mechanisms of programmed conversion from astrocytes to neurons have not yet been clarified.In the present study,we prepared reactive astrocytes from newborn rat spinal cord astrocytes using the scratch method and infected them with lentivirus carrying NEUROD1.The results showed that NEUROD1 overexpression reprogrammed the cultured reactive astrocytes into neurons in vitro with an efficiency of 13.4%.Using proteomic and bioinformatic analyses,1952 proteins were identified,of which 92 were differentially expressed.Among these proteins,11 were identified as candidate proteins in the process of reprogramming based on their biological functions and fold-changes in the bioinformatic analysis.Furthermore,western blot assay revealed that casein kinase II subunit alpha(CSNK2A2)and pinin(PNN)expression in NEUROD1-overexpressing reactive astrocytes was significantly increased,suggesting that NEUROD1 can directly reprogram spinal cord-derived reactive astrocytes into neurons in vitro,and that the NEUROD1-CSNK2A2-PNN pathway is involved in this process.This study was approved by the Animal Ethics Committee of Fujian Medical University,China(approval No.2016-05)on April 18,2016.