Molecular Characterization of Four ADF Genes Differentially Expressed in Cotton

Actin depolymerizing factor （ADF）, highly conserved in all eukaryotic cells, is a low molecular mass of actin-binding protein, which plays a key role in modulating the polymerizing and depolymerizing of the actin filaments. Four cDNAs （designated GhADF2, GhADF3, GhADF4, and GhADF5, respectively） encoding ADF proteins were isolated from cotton （Gossypium hirsutum） fiber cDNA library. GhADF2 cDNA is 705 bp in length and deduces a protein with 139 amino acids. GhADF3 cDNA is 819 bp in length and encodes a protein of 139 amino acids. GhADF4 cDNA is 804 bp in length and deduces a protein with 143 amino acids. GhADF5 cDNA is 644 bp in length and encodes a protein of 141 amino acids. The molecular evolutionary relationship of these genes was analyzed by means of bioinformatics. GhADF2 is closely related to GhADF3 （99% identity） and PetADF2 （89% identity）. GhADF4 is closely related to AtADF6 （78% identity）, and GhADF5 is closely related to AtADF5 （83% identity）. These results demonstrated that the plant ADF genes are highly conserved in structure. RT-PCR analysis showed that GhADF2 is predominantly expressed in fiber, whereas, GhADF5 is mainly expressed in cotyledons. On the other hand, it seems that GhADF3 and GhADF4 have no tissue specificity. Expression levels of different ADF genes may vary considerably in the same cell type, suggesting that they might be involved in regulating tissue development of cotton and the each ADF isoform may diverge to form the functional difference from the other ADFs during evolution.

Analysis of differential genes of ischemic stroke-induced heart tissue in mice based on GEO database

Objective:The differential genes of left ventricle in middle cerebral artery occlusion model(MCAO)mice and Sham mice(Sham)mice at 24h and 72h after ischemia were compared respectively,and the differential genes and their regulated functional pathways were analyzed at different time points after ischemic stroke,so as to analyze the mechanism of inducing cardiac dysfunction after ischemic stroke and provide evidence for its treatment.Methods:Gene-chip data from the left ventricle of MCAO mice and Sham mice were downloaded from the GEO database at the National Center for Biotechnology Information(NCBI).The differentially expressed genes were obtained by R language software programming.The GO functional enrichment and KEGG pathway enrichment analysis of the obtained differential genes were performed using DAVID 6.8 online analysis tool,and the Omicshare online analysis tool was used to visualize the enrichment analysis results.Results:At 24h after ischemia,187 differentially expressed genes were obtained,including 56 GO enrichment pathways and 5 KEGG enrichment pathways with significant significance.After 72h after ischemia,51 differentially expressed genes were obtained,14 GO enrichment pathways and 3 KEGG enrichment pathways with significant significance.The two time points involved Aplnr and Itgb6 gene targets and PI3K-Akt signaling pathway.Conclusion:①By analyzing the gene expression profile data,the differentially expressed genes and related pathways of cardiac dysfunction induced by ischemic stroke were obtained.②PI3K-AKT signaling pathway is closely related to the regulation of cardiac function,and regulation of PI3K-AKT signaling pathway may be an important direction for the treatment of cardiac dysfunction after ischemic stroke.

Cultivar Mixture Cooperation(CMC)of rice varieties(lines) with different blast resistant gene

We studied Cultivar Mixture Cooperation(CMC)of rice varieties(lines)by screening mixture ofrice varieties which possess different genes,1983-1996.

The Study on the Gene Expression of Preimplantation IVF Bovine Embryos

The cattle different stage embryos obtained from in vitro was studied using the technology of single preimplantation embryo mRNA different display:single 8-cell and blastocyst stage embryos were studied using technology of mRNA different display and one different fragment was found. The result suggested that this fragment displayed high homology (99%) to cattle mRNA for ribosomal protein L31. Then to detect the expression of RPL31mRNA in 8 cell and blastocyst stage embryos by real-time quantitative PCR,the result showed the relative amount of 8 cells was 3.2 times of blastocyst's.

Expression and analysis of zinc fi nger family gene in Lenzites gibbosa

Zinc finger transcription factors play significant roles in the growth and development of plant and animal,but their function remains obscure in fungi.Lenzites gibbosa mycelia were extracted and sequenced by transcriptome analysis after growing on sawdust at different times to support mycelial growth of L.gibbosa in a nutrient matrix.Data bases used for analysis were the Kyoto encyclopedia of genes and genomes(KEGG)annotation,the cluster of orthologous groups of proteins(COG)and gene ontology(GO)annotation.Zinc finger class genes related to the growth and development of L.gibbosa were screened.GO annotation and enrichment analysis of diff erentially expressed genes were carried out.A total of 114.55 Gb Clean Data were obtained from the L.gibbosa transcriptome.The average Clean Data in each sample was 6.16 Gb.The relative efficiency of reads between each sample and the reference genome was 88.5%to 91.4%.The COG analysis showed that most zinc finger protein genes were related to replication,recombination and repair function.GO enrichment analysis showed that the expressed genes involved in cellular process,cell part and binding.We identifi ed seventy-two expressed genes including seven up-regulated genes and sixty-five down-regulated genes by applying DESeq2 data analysis software.By comparing the significantly expressed genes with KEGG database,66 annotated sequences were obtained,and 35 primary metabolic pathways were annotated.Pathway enrichment analysis showed that differentially expressed genes were signifi cantly enriched in protein processing in endoplasmic reticulum and ubiquitin-mediated proteolysis pathways.Gene_11750 and gene_5266 are highly correlated with the growth and development of L.gibbosa and are closely related to protein processing in endoplasmic reticulum and ubiquitin-mediated proteolysis pathway.According to gene functional analysis,seven important differentially expressed genes related to the growth and development of L.gibbosa were identified.

Combining QTL mapping and expression profile analysis to identify candidate genes of cold tolerance from Dongxiang common wild rice(Oryza rufipogon Griff.)

Rice（Oryza sativa L.）, a tropical and subtropical crop, is susceptible to low temperature stress during seedling, booting, and flowering stages, which leads to lower grain quality levels and decreasing rice yields. Cold tolerance is affected by multiple genetic factors in rice, and the complex genetic mechanisms associated with chilling stress tolerance remain unclear. Here, we detected seven quantitative trait loci（QTLs） for cold tolerance at booting stage and identified one cold tolerant line, SIL157, in an introgression line population derived from a cross between the indica variety Guichao 2, as the recipient, and Dongxiang common wild rice, as the donor. When compared with Guichao 2, SIL157 showed a stronger cold tolerance during different growth stages. Through an integrated strategy that combined QTL-mapping with expression profile analysis, six candidate genes, which were up-regulated under chilling stress at the seedling and booting developmental stages, were studied. The results may help in understanding cold tolerance mechanisms and in using beneficial alleles from wild rice to improve the cold tolerance of rice cultivars through molecular marker-assisted selection.

Identification of key genes and related pathways in hepatocarcinoma usingbioinformatics analysis

Objective Various treatments have greatly reduced the mortality of hepatocellular carcinoma （HCC）. However, few therapies could be performed in advanced HCC. Therefore, understanding the characteristics of HCC at the level of the whole transcriptome can help prevent the progression of HCC. Methods： The aim of this study was to identify differently expressed genes and potent pathways between normal liver and HCC tissues. The gene expression profiles of GSE104627 were downloaded from Gene Expression Omnibus database. The Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses were performed and protein-protein interaction network of the differentially expressed genes were constructed by Cytoscape software. Results： In total, 880 differently expressed genes were identified between normal and tumor tissues, including 554 up-regulated genes and 326 down-regulated genes. Gene Ontology analysis results showed that the up-regulated genes were significantly enriched in establishment of RNA localization, nucleic acid transport, RNA transport, RNA localization and nucleobase, nucleoside, nucleotide and nucleic acid transport. Kyoto Encyclopedia of Genes and Genomes pathway analysis showed the up-regulated genes were enriched in axon guidance, dorso-ventral axis formation and pathways in cancer. The top 10 hub genes were identified from the protein - protein interaction network, and sub-networks revealed these genes were involved in significant pathways, including G protein-coupled receptors signaling pathway, signaling pathway via MAPK and extracellular matrix organization. Conclusion： The present study described the differently expressed genes between normal tissues and HCC tissues from the level of gene transcription. The possible signaling pathways involved in the development of HCC and related molecules involved were analyzed. However, further laboratory and clinical validation is still needed.

Cloning and Sequence Analysis of SLC11A1 Gene Promoter of Three Cattle Breeds in Xinjiang

[Objective]Solute carrier family 11 member 1（SLC11A1）is a major natural resistance candidate gene,which contributes to defense mechanisms of a variety of intracellular bacteria.The SLC11A1 gene promoter sequence of Xinjiang Brown Cattle,Holstein and Simmental were cloned in the test,and promoter sequence difference was analyzed,in order to provide genetic marker-assisted selection for disease-resistant breeding of dairy cattle.[Method]The Genomic DNA was extracted from whole blood collected from three cattle breeds in Xinjiang,and the 5’ flanking region of SLC11A1 gene was amplified by PCR and sequenced.The sequence was analyzed by bioinformatics software CpGplot,RepeatMasker,TFSEARCH,WWW Signal Scan and dual luciferase assay system.[Result]The SLC11A1 gene promoter sequence of 1 463 bp was confirmed,which had promoter activity.No CpG islands were found on promoter sequence.There were four different sites in SLC11A1 gene promoter sequences between Angus from America and three cattle breeds in Xinjiang.Sequence analysis revealed 12 transcription factor binding sites including Sp1,NF1,RelA-p65,GKLF,and CPBP.In promoter region there was an enhancer region（-734- -740）and two short scattered repetitive elements BOV-tA2,MIR3,as well as repeated DNA element Charlie8.[Conclusion]The SLC11A1 gene promoter sequences of three breeds were obtained,which were different from that of Angus.The paper provided a theoretical basis for further studying the influence of SLC11A1 gene polymorphisms on resistance against intracellular bacteria infection.

Genes Expressing Differences in Upland Cotton Fiber Development Between 12 DPA and 16 DPA

Cotton is the major dominant natural fiber crop on the earth.Although some current cotton genetics and breeding programs had made great progresses in cotton lint yield increases and disease resistance(tolerance),fiber quality has little improvement.Global understanding genes expression

The stability of E protein gene of the Japanese encephalitis live attenuated vaccine virus SA_(14)-14-2

The purpose of this study was to understand the stability and possibility of back mutation of Japanese encephalitis （JE） attenuated vaccine virus strain SA14-14-2 HKs on molecular level. The E genes of the SA14-14-2 HKs vaccine virus and its PHK cells passaged virus （SA14-14-2 HK17 ）, its mouse brain passaged virus （SA14-14-2 SM1 ） were sequenced and compared with the E gene of parental SAI4 virus. The total RNA was extracted from infected Vero cells and amplified by RT-PCR. The RT-PCR products were purified and cloned into T-vector. Positive clones were screened, identified and sequenced. There were twelve nucleotides and eight amino acids substitutions between SA14 parent virus and SA14-34-2 PHKs vaccine virus. The SA14-14-2 PHK17 virus showed two additional mutations （E-331 and E-398） which were not back mutations. Although five additional mutations were found in SA14-34-2 SMt virus, only one （E-307） was back mutation. Genetic characteristics of the attenuated vaccine virus SA14- 34-2 were stable when it was passaged 37 times on PHK cells or one time in mouse brains.

The first cavefish in the Dinaric Karst?Cave colonization made possible by phenotypic plasticity in Telestes karsticus

Cave animals are an excellent model system for studying adaptive evolution.At present,however,little is known about the mechanisms that enable surface colonizers to survive in the challenging environment of caves.One possibility is that these species have the necessary genetic background to respond with plastic changes to the pressures of underground habitats.To gain insight into this process,we conducted a comparative study with the fish species Telestes karsticus,which occurs in a hydrological system consisting of an interconnected stream and a cave.Results showed that T.karsticus resided year-round and spawned in Sušik cave,making it the first known cavefish in the Dinaric Karst.Cave and surface populations differed in morphological and physiological characteristics,as well as in patterns of gene expression without any evidence of genetic divergence.To test whether observed trait differences were plastic or genetic,we placed adult fish from both populations under light/dark or constant dark conditions.Common laboratory conditions erased all morphometric differences between the two morphs,suggesting phenotypic plasticity is driving the divergence of shape and size in wild fish.Lighter pigmentation and increased fat deposition exhibited by cave individuals were also observed in surface fish kept in the dark in the laboratory.Our study also revealed that specialized cave traits were not solely attributed to developmental plasticity,but also arose from adult responses,including acclimatization.Thus,we conclude that T.karsticus can adapt to cave conditions,with phenotypic plasticity playing an important role in the process of cave colonization.

C2C12成肌细胞体外诱导分化为肌管的实验

【目的】观察低浓度马血清体外诱导成肌细胞C2C12向成熟肌细胞分化、形成肌管的作用,探索定向诱导肌分化的途径及其分子机制。【方法】将C2C12成肌细胞用含不同浓度马血清的高糖DMEM培养基培养,收集细胞,在相差显微镜下观察形态变化,并通过免疫细胞化学(Imminocytochemistry,ICC)及RT-PCR方法分别检测肌性相关基因表达的改变。【结果】①20mL/L和50mL/L的马血清均能促使C2C12细胞形成肌小管,20mL/L浓度马血清诱导效率最好(第7天时肌小管形成率分别为31.4%±2.1%和19.0%±1.6%,P<0.001)。②20mL/L马血清诱导3～4d开始有肌管形成,之后肌管越来越多,到8～9d达高峰(第9天40.2%±1.3%),往后细胞逐渐衰老死亡。③20mL/L马血清诱导C2C12细胞向成熟肌细胞分化过程中,肌相关蛋白分子表达发生变化:结蛋白(desmin)、成肌分化抗原(MyoD)、生肌素(myogenin)和肌球蛋白(myosin)等表达逐渐增强(未刺激时分别约为49.6%±1.1%、23.4%±1.1%、4.8%±1.6%和2.6%±1.5%;第6天时分别提高为80.4%±1.8%、85.4%±1.1%、22.2%±1.1%和26.0%±1.6%;P<0.001);desmin、MyoD和myogenin相应的mRNA分子也可以通过RT-PCR检测到。【结论】①低浓度的马血清能够有效刺激C2C12成肌细胞向成熟细胞分化,以20mL/L浓度效果最好;②C2C12细胞是研究肌细胞发育分化的理想模板;③肌分化发育过程比较复杂,许多因素都可能起影响,研究设计要尽量标准化。

microRNA-425-5p对3T3-L1前体脂肪细胞增殖、分化的影响

为了探究miR-425-5p对小鼠3T3-L1前体脂肪细胞增殖、分化的影响效应,本试验采用实时荧光定量PCR检测miR-425-5p组织和细胞表达水平;运用CCK8、EdU、油红染色、甘油三酯含量分析等分别检测miR-425-5p对前体脂肪细胞增殖、分化的影响;利用生物信息学软件和双荧光素酶报告试验分别预测、验证miR-425-5p调控前体脂肪细胞分化的靶基因。结果表明,miR-425-5p在肥胖小鼠脂肪组织中低表达,在前体脂肪细胞增殖、分化过程中动态表达;与阴性对照相比,过表达miR-425-5p可促进前体脂肪细胞增殖,抑制脂肪细胞分化标志基因(PPARγ、C/EBPα、FAS等)表达,减少脂滴和甘油三酯积累;抑制miR-425-5p表达可抑制前体脂肪细胞增殖,阻止前体脂肪细胞诱导分化。在前体脂肪细胞分化过程中,过表达或抑制miR-425-5p可分别抑制或促进IGF1基因表达;与阴性对照相比,过表达miR-425-5p可抑制IGF1基因3′-UTR荧光活性,而突变miR-425-5p种子序列与IGF1基因3′-UTR的绑定位点可解除该抑制效果。综上所述,miR-425-5p可促进3T3-L1前体脂肪细胞增殖,并可直接靶向IGF1负向调控其分化。

酵母基因调控网络的微分方程模型研究

目的寻求一种新的描述基因调控网络的微分方程模型。方法建立微分方程并应用于酵母基因的时间序列表达数据,最后与试验结果比较。结果微分方程模型所得结论基本与试验结果吻合,并作出了预测。结论该模型弥补了其他线性微分方程模型的不足,是一种新的有价值的微分方程模型。

镁合金对骨髓间充质干细胞增殖及成骨分化的影响研究

目的探讨镁合金对人骨髓间充质干细胞(BMSCs)增殖及成骨分化的影响。方法抽取人骨BMSCs并予以鉴定;采用AZ31B镁合金浸提液培养人BMSCs 1、3、5、7 d,以普通培养基为对照组,CCK-8法检测细胞增殖活性;应用浸提液培养人BMSCs,诱导分化后检测成骨分化相关基因(COLⅠ、ALP、OPN)的表达。另设置调节pH值AZ31B组(pH-AZ31B组),以观察pH值变化对实验结果的影响。结果 BMSCs表达CD34(—)、CD45(—)、CD44(+)、CD73(+)、CD90(+)、CD105(+)。浸提液培养1、3、5、7 d,AZ31B组细胞相对增殖率低于对照组(P<0.05),毒性评级为2级;对照组和pH-AZ31B组细胞活性较好,两组细胞相对增殖率比较,差异无统计学意义(P>0.05)。AZ31B组COLⅠ基因表达量与对照组比较,差异无统计学意义(P>0.05);AZ31B组ALP基因表达量较低(P<0.05),pH-AZ31B组与对照组无显著差异(P>0.05);AZ31B组OPN基因表达量在诱导分化后6 d、12 d显著高于对照组(P<0.05),pH-AZ31B组与对照组无显著差异(P>0.05)。结论镁合金对BMSCs增殖及成骨分化无不良影响,其影响可能与浸提液pH值的变化有关。

藏、汉族中遗传差异与高原低氧适应研究

高海拔低氧是人类面临的严峻环境考验,容易引起高原病发生,而世居高原人群却对其具有独特的适应性。近年来有研究表明,缺氧诱导因子(hypoxia inducible factors,HIFs)相关的EPAS1、EGLN1/PHD2和HIF-1A基因多态性与表达在藏族、汉族人群中存在显著差异。对于这3种基因的深入研究有助于揭示藏族人适应高原的遗传机制。我们总结了近年来国内外关于EPAS1、EGLN1/PHD2和HIF-1A基因多态性、表达与高原低氧适应之间的关系。

甘氨酸-N-甲基转移酶对神经干细胞分化的影响

本文主要是研究甘氨酸鄄N鄄甲基转移酶(Gnm t)抑制后对神经干细胞分化的影响。从14d SD孕鼠的胚胎纹状体分离培养神经干细胞,培养两周后,将神经干细胞接种到24孔板,加入甘氨酸鄄N鄄甲基转移酶的产物SAH对该酶进行抑制,观察其抑制后对神经干细胞分化的影响。采用RT鄄PCR分析Gnm t抑制后m RNA的表达。结果表明:甘氨酸鄄N鄄甲基转移酶抑制可以诱导神经干细胞的分化。

Microarray Study of Mechanism of Trichostatin A Inducing Apoptosis of Molt-4 Cells

Histone deacetylase was overexpressed in a variety of cancers and was closely correlated with oncogenic factors. The histone deacetylase inhibitor, trichostatin A （TSA） was shown to induce apoptosis in many cancer cells. However, the mechanism of TSA on induction of cancer cells apoptosis is poorly understood. This study was designed to characterize the global gene expression profiles before and after treatment of human leukemia cell line Molt-4 with TSA. Flow cytometry, MTT and DNA ladder were used to observe the effect of TSA on the apoptosis of MOLT-4 cells and normal human peripheral blood mononuclear cells （PBMC）. Microarray, reverse transcription-polymerase chain reaction （RTopCR） and Western blotting were used to detect the difference of gene and protein expressions of Molt-4 cells after incubation of the cells with TSA. The results showed that TSA could induce Molt-4 apoptosis in dose- and time-dependent manners but spared PBMCs. Microarray analysis showed that after incubation with TSA for 9 h, 310 genes were upregulated and 313 genes were deregulated. These genes regulate the growth, differentiation and survival of cells. Among these genes, STATSA was down-regulated by 80.4% and MYC was down-regulated by 77.3%. It was concluded that TSA has definite growth-inhibiting and apoptosis-inducing effects on Molt-4 cells in time- and dose-dependent manners, with weak cytotoxic effects on PBMCs at the same time. The mechanism of TSA selectively inducing apoptosis and inhibiting growth may be ascribed to the changes of pro-proliferation genes and anti-apoptosis genes.

DIOXIN RECEPTOR GENE EXPRESSION IN DIFFERENT STAINS OF MICE

The aim of the present experiment is to find out whether there exist any differences in tissue expression of aryl

Relationship between FGF12 expression in high-grade gliomas and clinical features

Objective:gliomas are the most common intracranial tumors.Fibroblast growth factor-12(FGF12),which belongs to the fibroblast growth factor(FGFs)family,plays an important role in cell mitosis,as well as in other life functions,such as embryo development,tissue repair,cell proliferation,and tumor growth and invasion.The purpose of this study was to explore the potential value of FGF12 in high-grade gliomas and to predict its drug sensitivity.To provide a possible therapeutic target for glioma.Methods:high-grade glioma gene expression data and clinical information were downloaded from the gene expression omnibus(GEO)database,using the R language“impute”and“survival”survival analysis package.The FGF12 genes closely related to survival were screened,a survival curve was drawn,and clinical correlation analysis was conducted.The differentially expressed genes(DEGs)were defined as |logFC|>1,adj.PVal<0.05 as the standard.We used the David for Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)analysis,and constructed the protein-protein interactions(PPI)network.Then we used the Connectivity Map(CMAP)database for drug location,and the validation group was verified by the Chinese Glioma Genome Atlas(CGGA)database in the same way.Results:we found that high FGF12 expression was associated with a higher survival rate.The same validation was performed in the validation group through the CGGA database,and the survival curve showed the same trend.The expression level of FGF12 is an independent factor that affects the life time and status of the samples,and it is a low risk factor.GO enrichment analysis showed that differential genes were enriched in matrix transmembrane transporter activity,ion channels and calcium ion active channels.KEGG showed that DEGs were enriched in the phosphatidylinositol 3-kinase(PI3K)-protein kinase B(Akt)signaling pathway,dopaminergic synapse and cyclic adenosine monophosphate(cAMP)signaling pathway.Four seed genes,GRIA2,COLLA2,GRIA4 and HES6,were obtained by PPI network analysis.The cAMP was used to analyze and obtained 7 small molecule drugs,such as merbromin,naloxone,AH-2384&ticarcillin,vincamine,amoxicillin,azacyclonol,which may be helpful in the prognosis of high-grade gliomas.Conclusion:FGF12 and its pathway may serve as a biomarker or therapeutic target for high-grade gliomas.