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
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
Part 5＇ UTR region of pig SRPK1 gene was cloned by inverse PCR （I-PCR）, then a 425 bp gene sequence was acquired. A promoter region in -1--309 bp was predicted by online tool （TFSEARCH） and 32 binding sites of tr...Part 5＇ UTR region of pig SRPK1 gene was cloned by inverse PCR （I-PCR）, then a 425 bp gene sequence was acquired. A promoter region in -1--309 bp was predicted by online tool （TFSEARCH） and 32 binding sites of transcription with scores higher than 85 were getten, of which scores of Spl, MyoD, and HSF2 were over 90. Some of these binding sites of transcription factors were connected with promoters, but TATA-box, which was important to gene expression, hadn＇t been found in this region. By using PCR-SSCP method to search SNPs this part （5＇ UTR of SRPK1 in pig）, total of 40 Large White pigs were obtained as the research objects, but no polymorphism were found. Thus, 5＇ UTR of SRPK1 was speculated with a characteristic of high conservation, while it might have been directly or indirectly selected in commercial breeding. The paper provided a further feature of SRPK1 gene in molecular genetics.展开更多
TDF1(transcription-drived fragment) was homologous to the predicted S. lycopersicum nonspecific lipid-transfer protein,nsLTP 2-like(91%), and it was significantly upregulated in response to C. fulvum(cladosporium fulv...TDF1(transcription-drived fragment) was homologous to the predicted S. lycopersicum nonspecific lipid-transfer protein,nsLTP 2-like(91%), and it was significantly upregulated in response to C. fulvum(cladosporium fulvum) infection in tomato plants.In this experiment, the full-length cDNA of nsLTP 2-like was cloned using RACE technology based on the sequence of TDF1(GenBank: JZ717725). A full-length, 625 bp(GenBank: KU366289), cDNA sequence, which with 98% similarity to nsLTP 2-like gene(GenBank: XM015233692) was obtained. This cDNA contains an ORF(open reading frame) with full-length of 345 bp, coding of 114 amino acids, including 12.3% Ala and Gly. Protein molecular weight was 11.51 ku, the isoelectric point(pI) was 8.99, and average overall hydrophilicity was 0.412, with one phosphorylation sites, belonging to volatile acidic nuclear protein. Secondary structure prediction showed that α-Helix accounts for 30.7%, extension chain for 12.28%, β-corner for 9.65%, and random coil for 47.37%. Through comparative analysis of the homology among species, it was found that the amino acid sequence of tomato nsLTP 2-like protein had a high similarity with other plants, and with a specific conserved sequence which might related features in nsLTP 2-like protein. It also be analyzed the gene expression pattern of tomato in different parts and under different stress conditions.The results showed that nsLTP 2-like gene was up-regulated in varying degrees, under the condition of cold stress, exogenous hormone spraying and cladosporium fulvum infection. Therefore, it was speculated that the gene played a role in response to abiotic and biotic stress in tomato.展开更多
The nuclear receptor peroxisome proliferator-activated receptor γ（PPAR-γ） is a key transcriptional regulator of adipocyte differentiation.It also modulates the synthesis of adipocytokines in the adipose tissue.Its...The nuclear receptor peroxisome proliferator-activated receptor γ（PPAR-γ） is a key transcriptional regulator of adipocyte differentiation.It also modulates the synthesis of adipocytokines in the adipose tissue.Its polymorphisms are associated with the risk of type Ⅱ diabetes,obesity,cardiovascular diseases and cancer.In the present study,to investigate the regulatory mechanism of PPAR-γ gene on lipid metabolism,the computational prediction of peroxisome proliferator response elements（PPREs） was pursued with a genome-wide scale by using MEME/MAST method based on the information of TRANSFAC database,then GO and KEGG analyses were carried out.The results showed that a huge number of predicted target genes of PPAR-γ were significantly enriched in 36 GO terms（P〈0.05） and 10 KEGG pathways（P〈0.05） which were related closely to the lipid metabolism.The results should be a valuable resource for elucidation of the regulatory mechanism of PPAR-γ influence on lipid metabolism,also of the major importance to the diagnosis,prevention and treatment of the complex diseases such as obesity and diabete.展开更多
The regulating axillary branch gene was cloned and named as CsCCD7.Using a series bioinformatic computer softwares,database and online programes,CsCCD7 nucleotide sequence and CsCCD7 amino acid sequence were analyzed ...The regulating axillary branch gene was cloned and named as CsCCD7.Using a series bioinformatic computer softwares,database and online programes,CsCCD7 nucleotide sequence and CsCCD7 amino acid sequence were analyzed and CsCCD7 function was predicted.The results showed that CsCCD7 cDNA full length sequence was 2 136 bp,and included a 1 665 bp ORF which encoded a 554 AA protein;there were 32 kinds of cis-acting regulating element in 2 136 bp cDNA sequence;CsCCD7 was an unstable protein（the unstable coefficient：40.77）,including many phosphorylation sites related with CsCCD7 function;CsCCD7 had no transmembrane domain,and its subcellular localization was in chloroplast.CsCCD7 secondary structure contained four conformations including α-helix,β-sheet,β-turn and random coil.CsCCD7 protein had no signal peptide,so was non-secretory protein and hydrophilicity protein（grand average of hydropathicity：-0.401）.Both CsCCD7 secondary and tertiary structure prediction results showed that it was classified as carotenoid oxygenase family.Phylogenetic tree drew by Geneious showed that CsCCD7 was more closely related to AtCCD7 than any other Arabidopsis CCD protein.展开更多
Objective:Polycystic ovary syndrome(PCOS)is a common endocrine disorder that affects women’s health.This study aims to investigate gene and transcription factor(TF)expression differences between PCOS patients and hea...Objective:Polycystic ovary syndrome(PCOS)is a common endocrine disorder that affects women’s health.This study aims to investigate gene and transcription factor(TF)expression differences between PCOS patients and healthy individuals using bioinformatics approaches,and to verify the function of key transcription factors,with the goal of providing new insights into the pathogenesis of PCOS.Methods:Differentially expressed genes(DEGs)and differentially expressed transcription factors(DETFs)between PCOS patients and controls were identified from the RNA sequencing dataset GSE168404 using bioinformatics methods.Functional enrichment analysis was performed using Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)databases.The expression and function of core transcription factors were further validated in ovarian tissues of PCOS model mice and control mice using Western blotting and reverse transcription quantitative polymerase chain reaction(RTqPCR).Results:A total of 332 DEGs were identified between PCOS patients and controls,including 259 upregulated and 73 downregulated genes in the PCOS group.19 DETFs were further screened,of which 16 were upregulated and 3 were downregulated in PCOS.The upregulated DETFs(including TFCP2L1,DACH1,ESR2,AFF3,SMAD9,ZNF331,HOPX,ATOH8,HIF3α,DPF3,HOXC4,HES1,ID1,JDP2,SOX4,and ID3)were primarily associated with lipid metabolism,development,and cell adhesion.Protein and mRNA expression analysis in PCOS model mice revealed significantly decreased levels of hypoxia-inducible factor(HIF)1αand HIF2α,and significantly increased expression of HIF3αcompared to control mice(all P<0.001).Conclusion:Significant differences in gene and TF expression exist between PCOS patients and healthy individuals.HIF-3αmay play a crucial role in PCOS and could serve as a novel biomarker for diagnosis and a potential therapeutic target.展开更多
Potassium-calcium activates channel subfamily N member 3(KCNN3/SK3/KCa2.3)is involved in regulating cellular calcium signaling,muscle contraction and neurotransmitter release.Dysregulation of the KCNN3 channel is asso...Potassium-calcium activates channel subfamily N member 3(KCNN3/SK3/KCa2.3)is involved in regulating cellular calcium signaling,muscle contraction and neurotransmitter release.Dysregulation of the KCNN3 channel is associated with the development of various tumors.We use bioinformatics analysis to identify whether KCNN3 regulates the occurrence and development of stomach adenocarcinoma(STAD)as a prognostic target.By analyzing the Human Protein Atlas(HPA)database and The Cancer Genome Atlas(TCGA)database,we found that the protein and mRNA levels of KCNN3 were dramatically reduced in STAD,and TCGA database showed that KCNN3 significantly correlated with the prognosis and clinical features of STAD.In addition,we found that high expression of KCNN3 in STAD reduced the IC 50 of several drugs in STAD cells,suggesting that high expression of KCNN3 correlated with the drug sensitivity of STAD.To investigate the underlying biological mechanism,we identified a potential KCNN3 interaction factor,tumor necrosis factor receptor superfamily member 7(CD27/TNFRSF7),which is expressed at low levels in STAD.RT-qPCR and Western blotting confirmed that KCNN3 and CD27 positively correlated with each other at protein and mRNA levels,and co-immunoprecipitation and immunofluorescence experiments confirmed that the two proteins interact and colocalize in the cytoplasm.Moreover,we confirmed the inhibitory effect of KCNN3 on the proliferation,migration and invasion of human STAD cells in vitro and in vivo through subcutaneous tumorigenesis and cellular experiments.Furthermore,GO/KEGG enrichment analysis showed that KCNN3 was enriched in signaling pathways regulating the immune response and calcium or metal ion transport.Lastly,we verified through cell co-culture,RT-qPCR and CCK8 assays that high expression of KCNN3 can promote the increase of T cell activating factor and the killing effect of T cells on STAD cells.Therefore,our results suggest that KCNN3 is a potential inhibitory factor affecting the occurrence and progression of STAD.展开更多
Objective:Osteosarcoma is a highly aggressive primary malignant bone tumor commonly seen in children and adolescents,with a poor prognosis.Anchorage-dependent cell death(anoikis)has been proven to be indispensable in ...Objective:Osteosarcoma is a highly aggressive primary malignant bone tumor commonly seen in children and adolescents,with a poor prognosis.Anchorage-dependent cell death(anoikis)has been proven to be indispensable in tumor metastasis,regulating the migration and adhesion of tumor cells at the primary site.However,as a type of programmed cell death,anoikis is rarely studied in osteosarcoma,especially in the tumor immune microenvironment.This study aims to clarify prognostic value of anoikis and tumor immune microenvironment-related gene in the treatment of osteosarcoma.Methods:Anoikis-related genes(ANRGs)were obtained from GeneCards.Clinical information and ANRGs expression profiles of osteosarcoma patients were sourced from the therapeutically applicable research to generate effective therapies and Gene Expression Omnibus(GEO)databases.ANRGs highly associated with tumor immune microenvironment were identified by the estimate package and the weighted gene coexpression network analysis(WGCNA)algorithm.Machine learning algorithms were performed to construct long-term survival predictive strategy,each sample was divided into high-risk and low-risk subgroups,which was further verified in the GEO cohort.Finally,based on single-cell RNA-seq from the GEO database,analysis was done on the function of signature genes in the osteosarcoma tumor microenvironment.Results:A total of 51 hub ANRGs closely associated with the tumor microenvironment were identified,from which 3 genes(MERTK,BNIP3,S100A8)were selected to construct the prognostic model.Significant differences in immune cell activation and immune-related signaling pathways were observed between the high-risk and low-risk groups based on tumor microenvironment analysis(all P<0.05).Additionally,characteristic genes within the osteosarcoma microenvironment were identified in regulation of intercellular crosstalk through the GAS6-MERTK signaling pathway.Conclusion:The prognostic model based on ANRGs and tumor microenvironment demonstrate good predictive power and provide more personalized treatment options for patients with osteosarcoma.展开更多
Gene sequencing is a great way to interpret life, and high-throughput sequencing technology is a revolutionary technological innovation in gene sequencing researches. This technology is characterized by low cost and h...Gene sequencing is a great way to interpret life, and high-throughput sequencing technology is a revolutionary technological innovation in gene sequencing researches. This technology is characterized by low cost and high-throughput data. Currently, high-throughput sequencing technology has been widely applied in multi-level researches on genomics, transcriptomics and epigenomics. And it has fundamentally changed the way we approach problems in basic and translational researches and created many new possibilities. This paper presented a general description of high-throughput sequencing technology and a comprehensive review of its application with plain, concisely and precisely. In order to help researchers finish their work faster and better, promote science amateurs and understand it easier and better.展开更多
MicroRNAs, a class of endogenous non-coding small RNAs about 21 nucleotides in length, play pivotal roles in plant physiological and biochemical processes. Tomato is important economic crop throughout the world. Recen...MicroRNAs, a class of endogenous non-coding small RNAs about 21 nucleotides in length, play pivotal roles in plant physiological and biochemical processes. Tomato is important economic crop throughout the world. Recently, involvement of miRNAs in tomato has received significant attention. MiR172 is one of the conserved miRNAs in tomato. Investigation into the roles and the target genes of this small RNA molecular in Arabidopsis illustrated that miR172 functions in regulating the transitions between developmental stages and in specifying floral organ identify. Herein, we combined bioinformatics and molecular biology approaches to profile the functional characteristics of miR172 in tomato. The results of target prediction showed that AP2-like ethylene-responsive transcription factors were major targets of miR172, analysis of upstream sequence indicated the presence of stress-related cis-elements in its promoter regions, and further analysis of qRT-PCR confirmed that miR172 participated in various stress responses. Our research provided a paradigm for further in-depth investigation of the functional characteristics of miRNAs in tomato.展开更多
Background:GASA(Giberellic Acid Stimulated in Arabidopsis)gene family plays a crucial role in the phytohormone signaling pathway,growth and development,and stress responses in plants.Many GASA homologs have been ident...Background:GASA(Giberellic Acid Stimulated in Arabidopsis)gene family plays a crucial role in the phytohormone signaling pathway,growth and development,and stress responses in plants.Many GASA homologs have been identified in various plants.Nevertheless,little is known about these proteins in cotton.Results:In the current study,we identified 19,17,25,33,and 38 GASA genes via genome-wide analyses of Gossypium herbaceum,G.arboreum,G.raimondii,G.barbadense,and G.hirsutum,respectively,and performed comprehensive bioinformatics and expression analyses.According to our results,132 GASA proteins shared similar protein structures and were classified into four groups based on the phylogenetic tree.A synteny analysis suggested that segmental duplication was a key driver in the expansion of the GASA gene family.Meanwhile,the cis-element and protein interaction analyses indicated that GhGASA proteins play a significant role in the hormone responses.Transcriptomic and qRT-PCR(Quantitative real time-polymerase chain reaction)analyses revealed diverse expression profiles of the GhGASA genes in different organs under abiotic stresses,indicating that some GhGASA genes possibly participate in fiber development and abiotic-stress responses.Conclusions:The GASA genes in cotton were systematically identified and analyzed for the first time in this paper,and it suggested that the GASA genes are important to the development and growth of cotton.These results will support future exploration of the functions of GASA genes in cotton.展开更多
The full-length cDNA of 909 bp of the osteopontin gene(OPN) was isolated from of Large White Pig and analyzed with bioinformatics methods.The results showed high proportions of Asp,Glu and Ser and verified presence of...The full-length cDNA of 909 bp of the osteopontin gene(OPN) was isolated from of Large White Pig and analyzed with bioinformatics methods.The results showed high proportions of Asp,Glu and Ser and verified presence of the special sequence Arg-Gly-Asp(RGD) in the primary structure of OPN.There were high proportions of alpha helices and strong hydrophilicity in the secondary structure. The signature sequence of OPN([KQ]-x-[TA]-x(2)-[GA]-S-S-E-E-K) was located in the first region of high homology.Two phylogenetic trees were constructed,based on the entire OPN protein sequence and the conserved signature sequence,and showed that the relationship between pig and cow was the closest, but farthest between pig and chicken.OPN mRNA was expressed in many tissues of the pig:higher in the stomach,kidney,lung,small intestine and ovary,and lower in the heart,spleen and large intestine.The OPN protein size differed in different tissues:70 kDa in liver and muscle,70 and 45 kDa in stomach,small intestines and kidney,70,45 and 24 kDa in lung,heart and uterus.展开更多
Structure-based protein classification can be based on the similarities in primary, second or tertiary structures of proteins. A method using virtual-bond-angles series that transformed the protein space configuration...Structure-based protein classification can be based on the similarities in primary, second or tertiary structures of proteins. A method using virtual-bond-angles series that transformed the protein space configuration into a sequence was used for the classification of three-dimensional structures oi proteins. By transforming the main chains formed by C^a atoms of proteins into sequences, the series of virtual-bond-angles corresponding to the tertiary structure of the proteins were constructed. Then a distance-based hierarchical clustering method similar to Ward method was introduced to classify these virtual-bond-angles series of proteins. 200 files of protein structures were selected from Brookheaven protein data bank, and 11 clusters were classified.展开更多
In this study,a rapid molecular marking method,gspCAPS（genome sequence pool CAPS）,was established to develop soybean CAPS markers.The bioinformatics tools were executed for analyzing the sequences of soybean SNP sit...In this study,a rapid molecular marking method,gspCAPS（genome sequence pool CAPS）,was established to develop soybean CAPS markers.The bioinformatics tools were executed for analyzing the sequences of soybean SNP sites and selecting the appropriate restriction endonucleases.Meanwhile,DNA samples were mixed,and the primers were sequentially screened by PCR and enzyme digestion procedures.To evaluate this method,61 pairs of primers were designed and polymorphisms were detected in nine soybean varieties.The results showed that the method remarkably improved its efficiency and characterized with advantages such as high efficiency,timesaving and low cost,which had a widespread prospect for applying in the future.展开更多
A software and algorithm which based on random sequence model uses osmotic stress responding cis elements from existing information sources of biology was designed. It can infer the genic downstream function of Arabid...A software and algorithm which based on random sequence model uses osmotic stress responding cis elements from existing information sources of biology was designed. It can infer the genic downstream function of Arabidopsis thaliana through analyzing its promoter region, and can offer effective aided analysis to mine osmotic stress responding genes in Arabidopsis thatiana genome. The practical application proves that this software can aid to analyze vast genic data and offer important data evidence.展开更多
Arabidopsis thaliana ovate family proteins （AtOFPs） is a newly found plant-specific protein family interacting with TALE （3-aa loop extension homeodomain proteins） homeodomain proteins in Arabidopsis. Here, based ...Arabidopsis thaliana ovate family proteins （AtOFPs） is a newly found plant-specific protein family interacting with TALE （3-aa loop extension homeodomain proteins） homeodomain proteins in Arabidopsis. Here, based on bioinformatic analysis, we found that Arabidopsis genome actually encoded 17 OVATE domain-containing proteins. One of them, AtOFP19, has not been previously identified. Based on their amino acid sequence similarity, AtOFPs proteins can be divided into two groups. Most of the AtOFPs were located in nuclear, four of them were presented in chloroplast and the remaining two members appeared in cytoplasmic. A genome- wide microarray based gene expression analysis involving 47 stages of vegetative and reproductive development revealed that AtOFPs have diverse expression pattems. Investigation of proteins interaction showed that nine AtOFPs only interacted with TALE homeodomain proteins, which are fundamental regulators of plant meristem function and leaf development. Our work could provide important leads toward functional genomics studies of ovate family proteins, which may be involved in a previously unrecognized control mechanism in plant development展开更多
The p65 protein is a functional subunit of NF-κB family and exhibits a crucial role in host immune and inflammatory responses,apoptosis and tumor proliferation if improperly-regulated.Given its ubiquitous association...The p65 protein is a functional subunit of NF-κB family and exhibits a crucial role in host immune and inflammatory responses,apoptosis and tumor proliferation if improperly-regulated.Given its ubiquitous association with nearly all the animal cells and its pleotropic functions,the gene encoding NF-κB p65 subunit was cloned and sequenced from porcine kidney(PK-15)cells.The gene was 1662 bp in length,encoded a 553-amino acid protein and contained the prototypical NF-κB functional domains.Real-time quantitative RT-PCR and Western blot were used to characterize the transcription and expression levels of the p65 in different pig tissues.The results indicated that the p65 gene and protein were both broadly expressed in pig tissues,but most highly expressed in the intestine-associated lymph nodes and the lungs.To localize the recombinant protein in intestinal porcine epithelial cells(IPEC-J2),the gene was subcloned into the vector pEGFP(pEGFP-p65).Using fluorescence microscopy,the protein was found confined to the cytoplasm in normal cells;however,during porcine epidemic diarrhea virus(PEDV)infection,mRNA and protein expression were significantly up-regulated and the protein exhibited an overt tendency for nuclear translocalization consistent with a regulatory role in antiviral innate immunity.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 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 11th Five-year Plan Key Projects of National Science and Technology (2008BADB2B01)
文摘Part 5＇ UTR region of pig SRPK1 gene was cloned by inverse PCR （I-PCR）, then a 425 bp gene sequence was acquired. A promoter region in -1--309 bp was predicted by online tool （TFSEARCH） and 32 binding sites of transcription with scores higher than 85 were getten, of which scores of Spl, MyoD, and HSF2 were over 90. Some of these binding sites of transcription factors were connected with promoters, but TATA-box, which was important to gene expression, hadn＇t been found in this region. By using PCR-SSCP method to search SNPs this part （5＇ UTR of SRPK1 in pig）, total of 40 Large White pigs were obtained as the research objects, but no polymorphism were found. Thus, 5＇ UTR of SRPK1 was speculated with a characteristic of high conservation, while it might have been directly or indirectly selected in commercial breeding. The paper provided a further feature of SRPK1 gene in molecular genetics.
基金Supported by the National Key R&D Program of China(2017YFD0101900)China Agriculture Research System(CARS-23-A-16)the Science Foundation of Heilongjiang Province(C2017024)
文摘TDF1(transcription-drived fragment) was homologous to the predicted S. lycopersicum nonspecific lipid-transfer protein,nsLTP 2-like(91%), and it was significantly upregulated in response to C. fulvum(cladosporium fulvum) infection in tomato plants.In this experiment, the full-length cDNA of nsLTP 2-like was cloned using RACE technology based on the sequence of TDF1(GenBank: JZ717725). A full-length, 625 bp(GenBank: KU366289), cDNA sequence, which with 98% similarity to nsLTP 2-like gene(GenBank: XM015233692) was obtained. This cDNA contains an ORF(open reading frame) with full-length of 345 bp, coding of 114 amino acids, including 12.3% Ala and Gly. Protein molecular weight was 11.51 ku, the isoelectric point(pI) was 8.99, and average overall hydrophilicity was 0.412, with one phosphorylation sites, belonging to volatile acidic nuclear protein. Secondary structure prediction showed that α-Helix accounts for 30.7%, extension chain for 12.28%, β-corner for 9.65%, and random coil for 47.37%. Through comparative analysis of the homology among species, it was found that the amino acid sequence of tomato nsLTP 2-like protein had a high similarity with other plants, and with a specific conserved sequence which might related features in nsLTP 2-like protein. It also be analyzed the gene expression pattern of tomato in different parts and under different stress conditions.The results showed that nsLTP 2-like gene was up-regulated in varying degrees, under the condition of cold stress, exogenous hormone spraying and cladosporium fulvum infection. Therefore, it was speculated that the gene played a role in response to abiotic and biotic stress in tomato.
基金Supported by National 973 Project of China (2006CB102105)Natural Science Foundation Key Project of Heilongjiang Province (ZJN0604-01)National 863 Project of China (2006AA10A120)
文摘The nuclear receptor peroxisome proliferator-activated receptor γ（PPAR-γ） is a key transcriptional regulator of adipocyte differentiation.It also modulates the synthesis of adipocytokines in the adipose tissue.Its polymorphisms are associated with the risk of type Ⅱ diabetes,obesity,cardiovascular diseases and cancer.In the present study,to investigate the regulatory mechanism of PPAR-γ gene on lipid metabolism,the computational prediction of peroxisome proliferator response elements（PPREs） was pursued with a genome-wide scale by using MEME/MAST method based on the information of TRANSFAC database,then GO and KEGG analyses were carried out.The results showed that a huge number of predicted target genes of PPAR-γ were significantly enriched in 36 GO terms（P〈0.05） and 10 KEGG pathways（P〈0.05） which were related closely to the lipid metabolism.The results should be a valuable resource for elucidation of the regulatory mechanism of PPAR-γ influence on lipid metabolism,also of the major importance to the diagnosis,prevention and treatment of the complex diseases such as obesity and diabete.
基金Supported by Doctoral Fund of Ministry of Education of China (200802240008)National 863 Project (2007AA10Z177)
文摘The regulating axillary branch gene was cloned and named as CsCCD7.Using a series bioinformatic computer softwares,database and online programes,CsCCD7 nucleotide sequence and CsCCD7 amino acid sequence were analyzed and CsCCD7 function was predicted.The results showed that CsCCD7 cDNA full length sequence was 2 136 bp,and included a 1 665 bp ORF which encoded a 554 AA protein;there were 32 kinds of cis-acting regulating element in 2 136 bp cDNA sequence;CsCCD7 was an unstable protein（the unstable coefficient：40.77）,including many phosphorylation sites related with CsCCD7 function;CsCCD7 had no transmembrane domain,and its subcellular localization was in chloroplast.CsCCD7 secondary structure contained four conformations including α-helix,β-sheet,β-turn and random coil.CsCCD7 protein had no signal peptide,so was non-secretory protein and hydrophilicity protein（grand average of hydropathicity：-0.401）.Both CsCCD7 secondary and tertiary structure prediction results showed that it was classified as carotenoid oxygenase family.Phylogenetic tree drew by Geneious showed that CsCCD7 was more closely related to AtCCD7 than any other Arabidopsis CCD protein.
基金supported by the Natural Science Foundation of Hunan Province,China(2022JJ30886).
文摘Objective:Polycystic ovary syndrome(PCOS)is a common endocrine disorder that affects women’s health.This study aims to investigate gene and transcription factor(TF)expression differences between PCOS patients and healthy individuals using bioinformatics approaches,and to verify the function of key transcription factors,with the goal of providing new insights into the pathogenesis of PCOS.Methods:Differentially expressed genes(DEGs)and differentially expressed transcription factors(DETFs)between PCOS patients and controls were identified from the RNA sequencing dataset GSE168404 using bioinformatics methods.Functional enrichment analysis was performed using Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)databases.The expression and function of core transcription factors were further validated in ovarian tissues of PCOS model mice and control mice using Western blotting and reverse transcription quantitative polymerase chain reaction(RTqPCR).Results:A total of 332 DEGs were identified between PCOS patients and controls,including 259 upregulated and 73 downregulated genes in the PCOS group.19 DETFs were further screened,of which 16 were upregulated and 3 were downregulated in PCOS.The upregulated DETFs(including TFCP2L1,DACH1,ESR2,AFF3,SMAD9,ZNF331,HOPX,ATOH8,HIF3α,DPF3,HOXC4,HES1,ID1,JDP2,SOX4,and ID3)were primarily associated with lipid metabolism,development,and cell adhesion.Protein and mRNA expression analysis in PCOS model mice revealed significantly decreased levels of hypoxia-inducible factor(HIF)1αand HIF2α,and significantly increased expression of HIF3αcompared to control mice(all P<0.001).Conclusion:Significant differences in gene and TF expression exist between PCOS patients and healthy individuals.HIF-3αmay play a crucial role in PCOS and could serve as a novel biomarker for diagnosis and a potential therapeutic target.
文摘Potassium-calcium activates channel subfamily N member 3(KCNN3/SK3/KCa2.3)is involved in regulating cellular calcium signaling,muscle contraction and neurotransmitter release.Dysregulation of the KCNN3 channel is associated with the development of various tumors.We use bioinformatics analysis to identify whether KCNN3 regulates the occurrence and development of stomach adenocarcinoma(STAD)as a prognostic target.By analyzing the Human Protein Atlas(HPA)database and The Cancer Genome Atlas(TCGA)database,we found that the protein and mRNA levels of KCNN3 were dramatically reduced in STAD,and TCGA database showed that KCNN3 significantly correlated with the prognosis and clinical features of STAD.In addition,we found that high expression of KCNN3 in STAD reduced the IC 50 of several drugs in STAD cells,suggesting that high expression of KCNN3 correlated with the drug sensitivity of STAD.To investigate the underlying biological mechanism,we identified a potential KCNN3 interaction factor,tumor necrosis factor receptor superfamily member 7(CD27/TNFRSF7),which is expressed at low levels in STAD.RT-qPCR and Western blotting confirmed that KCNN3 and CD27 positively correlated with each other at protein and mRNA levels,and co-immunoprecipitation and immunofluorescence experiments confirmed that the two proteins interact and colocalize in the cytoplasm.Moreover,we confirmed the inhibitory effect of KCNN3 on the proliferation,migration and invasion of human STAD cells in vitro and in vivo through subcutaneous tumorigenesis and cellular experiments.Furthermore,GO/KEGG enrichment analysis showed that KCNN3 was enriched in signaling pathways regulating the immune response and calcium or metal ion transport.Lastly,we verified through cell co-culture,RT-qPCR and CCK8 assays that high expression of KCNN3 can promote the increase of T cell activating factor and the killing effect of T cells on STAD cells.Therefore,our results suggest that KCNN3 is a potential inhibitory factor affecting the occurrence and progression of STAD.
基金This work was supported by the National Natural Science Foundation(82172594 and 82373046)the Hunan Graduate Research Innovation Project(CX20230318),China.
文摘Objective:Osteosarcoma is a highly aggressive primary malignant bone tumor commonly seen in children and adolescents,with a poor prognosis.Anchorage-dependent cell death(anoikis)has been proven to be indispensable in tumor metastasis,regulating the migration and adhesion of tumor cells at the primary site.However,as a type of programmed cell death,anoikis is rarely studied in osteosarcoma,especially in the tumor immune microenvironment.This study aims to clarify prognostic value of anoikis and tumor immune microenvironment-related gene in the treatment of osteosarcoma.Methods:Anoikis-related genes(ANRGs)were obtained from GeneCards.Clinical information and ANRGs expression profiles of osteosarcoma patients were sourced from the therapeutically applicable research to generate effective therapies and Gene Expression Omnibus(GEO)databases.ANRGs highly associated with tumor immune microenvironment were identified by the estimate package and the weighted gene coexpression network analysis(WGCNA)algorithm.Machine learning algorithms were performed to construct long-term survival predictive strategy,each sample was divided into high-risk and low-risk subgroups,which was further verified in the GEO cohort.Finally,based on single-cell RNA-seq from the GEO database,analysis was done on the function of signature genes in the osteosarcoma tumor microenvironment.Results:A total of 51 hub ANRGs closely associated with the tumor microenvironment were identified,from which 3 genes(MERTK,BNIP3,S100A8)were selected to construct the prognostic model.Significant differences in immune cell activation and immune-related signaling pathways were observed between the high-risk and low-risk groups based on tumor microenvironment analysis(all P<0.05).Additionally,characteristic genes within the osteosarcoma microenvironment were identified in regulation of intercellular crosstalk through the GAS6-MERTK signaling pathway.Conclusion:The prognostic model based on ANRGs and tumor microenvironment demonstrate good predictive power and provide more personalized treatment options for patients with osteosarcoma.
基金Supported by the National Natural Science Foundations of China(3127218631301791)
文摘Gene sequencing is a great way to interpret life, and high-throughput sequencing technology is a revolutionary technological innovation in gene sequencing researches. This technology is characterized by low cost and high-throughput data. Currently, high-throughput sequencing technology has been widely applied in multi-level researches on genomics, transcriptomics and epigenomics. And it has fundamentally changed the way we approach problems in basic and translational researches and created many new possibilities. This paper presented a general description of high-throughput sequencing technology and a comprehensive review of its application with plain, concisely and precisely. In order to help researchers finish their work faster and better, promote science amateurs and understand it easier and better.
基金Suppported by the National Natural Science Foundation of China(31171971,31272167)
文摘MicroRNAs, a class of endogenous non-coding small RNAs about 21 nucleotides in length, play pivotal roles in plant physiological and biochemical processes. Tomato is important economic crop throughout the world. Recently, involvement of miRNAs in tomato has received significant attention. MiR172 is one of the conserved miRNAs in tomato. Investigation into the roles and the target genes of this small RNA molecular in Arabidopsis illustrated that miR172 functions in regulating the transitions between developmental stages and in specifying floral organ identify. Herein, we combined bioinformatics and molecular biology approaches to profile the functional characteristics of miR172 in tomato. The results of target prediction showed that AP2-like ethylene-responsive transcription factors were major targets of miR172, analysis of upstream sequence indicated the presence of stress-related cis-elements in its promoter regions, and further analysis of qRT-PCR confirmed that miR172 participated in various stress responses. Our research provided a paradigm for further in-depth investigation of the functional characteristics of miRNAs in tomato.
基金the National Natural Science Foundation of China(grant no.31701474).
文摘Background:GASA(Giberellic Acid Stimulated in Arabidopsis)gene family plays a crucial role in the phytohormone signaling pathway,growth and development,and stress responses in plants.Many GASA homologs have been identified in various plants.Nevertheless,little is known about these proteins in cotton.Results:In the current study,we identified 19,17,25,33,and 38 GASA genes via genome-wide analyses of Gossypium herbaceum,G.arboreum,G.raimondii,G.barbadense,and G.hirsutum,respectively,and performed comprehensive bioinformatics and expression analyses.According to our results,132 GASA proteins shared similar protein structures and were classified into four groups based on the phylogenetic tree.A synteny analysis suggested that segmental duplication was a key driver in the expansion of the GASA gene family.Meanwhile,the cis-element and protein interaction analyses indicated that GhGASA proteins play a significant role in the hormone responses.Transcriptomic and qRT-PCR(Quantitative real time-polymerase chain reaction)analyses revealed diverse expression profiles of the GhGASA genes in different organs under abiotic stresses,indicating that some GhGASA genes possibly participate in fiber development and abiotic-stress responses.Conclusions:The GASA genes in cotton were systematically identified and analyzed for the first time in this paper,and it suggested that the GASA genes are important to the development and growth of cotton.These results will support future exploration of the functions of GASA genes in cotton.
基金supported by Scientific and Technological Project in Heilongjiang Province (GB05B 106)Postdoctoral Fund of Heilongjiang Province(LRB07-392)
文摘The full-length cDNA of 909 bp of the osteopontin gene(OPN) was isolated from of Large White Pig and analyzed with bioinformatics methods.The results showed high proportions of Asp,Glu and Ser and verified presence of the special sequence Arg-Gly-Asp(RGD) in the primary structure of OPN.There were high proportions of alpha helices and strong hydrophilicity in the secondary structure. The signature sequence of OPN([KQ]-x-[TA]-x(2)-[GA]-S-S-E-E-K) was located in the first region of high homology.Two phylogenetic trees were constructed,based on the entire OPN protein sequence and the conserved signature sequence,and showed that the relationship between pig and cow was the closest, but farthest between pig and chicken.OPN mRNA was expressed in many tissues of the pig:higher in the stomach,kidney,lung,small intestine and ovary,and lower in the heart,spleen and large intestine.The OPN protein size differed in different tissues:70 kDa in liver and muscle,70 and 45 kDa in stomach,small intestines and kidney,70,45 and 24 kDa in lung,heart and uterus.
文摘Structure-based protein classification can be based on the similarities in primary, second or tertiary structures of proteins. A method using virtual-bond-angles series that transformed the protein space configuration into a sequence was used for the classification of three-dimensional structures oi proteins. By transforming the main chains formed by C^a atoms of proteins into sequences, the series of virtual-bond-angles corresponding to the tertiary structure of the proteins were constructed. Then a distance-based hierarchical clustering method similar to Ward method was introduced to classify these virtual-bond-angles series of proteins. 200 files of protein structures were selected from Brookheaven protein data bank, and 11 clusters were classified.
基金Supported by National High-Tech Research Program (2006AA100104, 2008AA10Z153)Innovation Research Group of NEAU (CXT004)
文摘In this study,a rapid molecular marking method,gspCAPS（genome sequence pool CAPS）,was established to develop soybean CAPS markers.The bioinformatics tools were executed for analyzing the sequences of soybean SNP sites and selecting the appropriate restriction endonucleases.Meanwhile,DNA samples were mixed,and the primers were sequentially screened by PCR and enzyme digestion procedures.To evaluate this method,61 pairs of primers were designed and polymorphisms were detected in nine soybean varieties.The results showed that the method remarkably improved its efficiency and characterized with advantages such as high efficiency,timesaving and low cost,which had a widespread prospect for applying in the future.
文摘A software and algorithm which based on random sequence model uses osmotic stress responding cis elements from existing information sources of biology was designed. It can infer the genic downstream function of Arabidopsis thaliana through analyzing its promoter region, and can offer effective aided analysis to mine osmotic stress responding genes in Arabidopsis thatiana genome. The practical application proves that this software can aid to analyze vast genic data and offer important data evidence.
基金Supported by the National Natural Science Foundation of China (30870144)
文摘Arabidopsis thaliana ovate family proteins （AtOFPs） is a newly found plant-specific protein family interacting with TALE （3-aa loop extension homeodomain proteins） homeodomain proteins in Arabidopsis. Here, based on bioinformatic analysis, we found that Arabidopsis genome actually encoded 17 OVATE domain-containing proteins. One of them, AtOFP19, has not been previously identified. Based on their amino acid sequence similarity, AtOFPs proteins can be divided into two groups. Most of the AtOFPs were located in nuclear, four of them were presented in chloroplast and the remaining two members appeared in cytoplasmic. A genome- wide microarray based gene expression analysis involving 47 stages of vegetative and reproductive development revealed that AtOFPs have diverse expression pattems. Investigation of proteins interaction showed that nine AtOFPs only interacted with TALE homeodomain proteins, which are fundamental regulators of plant meristem function and leaf development. Our work could provide important leads toward functional genomics studies of ovate family proteins, which may be involved in a previously unrecognized control mechanism in plant development
基金Supported by the National Natural Science Foundation of China(31370140,31372438)。
文摘The p65 protein is a functional subunit of NF-κB family and exhibits a crucial role in host immune and inflammatory responses,apoptosis and tumor proliferation if improperly-regulated.Given its ubiquitous association with nearly all the animal cells and its pleotropic functions,the gene encoding NF-κB p65 subunit was cloned and sequenced from porcine kidney(PK-15)cells.The gene was 1662 bp in length,encoded a 553-amino acid protein and contained the prototypical NF-κB functional domains.Real-time quantitative RT-PCR and Western blot were used to characterize the transcription and expression levels of the p65 in different pig tissues.The results indicated that the p65 gene and protein were both broadly expressed in pig tissues,but most highly expressed in the intestine-associated lymph nodes and the lungs.To localize the recombinant protein in intestinal porcine epithelial cells(IPEC-J2),the gene was subcloned into the vector pEGFP(pEGFP-p65).Using fluorescence microscopy,the protein was found confined to the cytoplasm in normal cells;however,during porcine epidemic diarrhea virus(PEDV)infection,mRNA and protein expression were significantly up-regulated and the protein exhibited an overt tendency for nuclear translocalization consistent with a regulatory role in antiviral innate immunity.
