The fall armyworm, Spodoptera frugiperda, has developed extremely high levels of resistance to chlorantraniliprole and other classes of insecticides in the field. As microRNAs (miRNAs) play important roles in various biological processes through gene regulation. we examined the miRNA profile of S. frugiperda in response to Chlorantraniliprole, Indoxacarb and Thiamethoxam. Transcriptome analysis showed significant changes in the abundance of some miRNAs after treatment of S. frugiperda larvae with LC20 concentrations of three insecticides. A total of 197 miRNAs were systematically identified from S. frugiperda, and 16, 9, 2 miRNAs were differentially expressed after treatments of three insecticides. Importantly, three miRNAs were significantly downregulated and three were upregulated by RT-qPCR after treatment the LC50 of three insecticides with S. frugiperda larvae. Microinjection of agomirs of these six miRNAs into S. frugiperda larvae resulted in significant changes in mortality rates when exposed to three insecticides. Additionally, we also screened potential target genes for some of differentially expressed miRNAs, which may play important roles in insecticide resistance development. These findings provide valuable insights into the molecular mechanisms of insecticide resistance and underscore the potential of miRNAs as targets for the development of novel pest control strategies in S. frugiperda.

A new fumigant, carbonyl sulfide (COS), has potential for use as a replacement for methyl bromide, yet its mechanism of toxicity to insects remains poorly understood. In this study, transcriptome analysis was performed on Tribolium castaneum malpighian tubules and fat bodies, which are known to play an essential role in energy storage and utilization in insect species. In total, upon exposure to COS, 3,034 and 2,973 genes were differentially expressed in the T. castaneum malpighian tubules and fat body, respectively. These differentially expressed genes comprise a significant number of detoxification-related genes, including 105 P450s, 18 glutathione S-transferases (GSTs), 82 ABC transporters, 25 UDP-glucosyltransferases and 42 carboxylesterases and mitochondrial–related genes, including 9 complex Ⅰ genes, 2 complex Ⅱ genes, 1 complex Ⅲ gene, 9 complex IV genes, 8 complex V genes from both malpighian tubules and fat body tissues. Moreover, KEGG analysis demonstrated that the upregulated genes were enriched in xenobiotic metabolism by ABC transporters and drug metabolism by other enzymes. We also investigated the role of carbonic anhydrases (CAs) in toxicity of COS using dsRNA treatment in T. castaneum. These results show that CA genes have a key role in toxicity of the COS. Furthermore, the results of transcriptomic analysis provide new insights into the insecticidal mechanism of COS fumigation against T. castaneum and eventually contribute to the management of this important stored grain pests.

Colon cancer is one of the most common malignant tumors, but there are still a few validated biomarkers of colon cancer. Exosome-mediated microRNAs (miRNAs) have been recognized as potential biomarkers in cancers, and miRNAs can regulate a variety of genes. Recently, Fusobacterium nucleatum was discovered in the tissues of human colon cancer patients. Its role in colon cancer was highlighted. F. nucleatum may contribute to the progression of colon cancer through the mechanism of exosome-mediated miRNAs transfer. However, the exosomal miRNAs regulation mechanism by F. nucleatum in colon cancer is not well known. Thus, we performed next-generation sequencing to investigate the overall pattern of exosomal miRNAs expression in the colon cancer cell culture supernatant. We have confirmed the alterations of various exosomal miRNAs. In addition, to investigate the function of exosomal miRNAs, a Kyoto Encyclopedia of Genes and Genomes analysis was performed on the target genes of changed miRNAs. Potential target genes were associated with a variety of signaling pathways, and one of these pathways was related to colorectal cancer. These findings suggested that F. nucleatum can alter exosomal miRNAs released from colorectal cancer cells. Furthermore, exosomal miRNAs altered by F. nucleatum could be potential biomarkers for the diagnosis and therapy of colon cancer.

Exosomes are Nano-sized lipid vesicles secreted from mammalian cells containing diverse cellular materials such as proteins, lipids, and nucleotides. Multiple lines of evidence indicate that in saliva, exosomes and their contents such as microRNAs (miRNAs) mediate numerous cellular responses upon delivery to recipient cells. The objective of this study was to characterize the different expression profile of exosomal miRNAs in saliva samples, periodically isolated from a single periodontitis patient. Unstimulated saliva was collected from a single patient over time periods for managing periodontitis. MicroRNAs extracted from each phase were investigated for the expression of exosomal miRNAs. Salivary exosomal miRNAs were analyzed using Affymetrix miRNA arrays and prediction of target genes and pathways for its different expression performed using DIANA-mirPath, a web-based, computational tool. Following the delivery of miRNA mimics (hsa-miR-4487, -4532, and -7108-5p) into human gingival fibroblasts, the expression of pro-inflammatory cytokines and activation of the MAPK pathway were evaluated through RT-PCR and western blotting. In each phase, 13 and 43 miRNAs were found to be differently expressed (|FC| ≥ 2). Among these, hsa-miR-4487 (|FC|=9.292005) and hasmiR- 4532 (|FC|=18.322697) were highly up-regulated in the clinically severe phase, whereas hsa-miR-7108-5p (|FC|= 12.20601) was strongly up-regulated in the clinically mild phase. In addition, the overexpression of miRNA mimics in human gingival fibroblasts resulted in a significant induction of IL-6 mRNA expression and p38 phosphorylation. The findings of this study established alterations in salivary exosomal miRNAs which are dependent on the severity of periodontitis and may act as potential candidates for the treatment of oral inflammatory diseases.

Odontogenic cells express many genes spatiotemporally through complex and intricate processes during tooth formation. Therefore, investigating them during the tooth development has been an important subject for the better understanding of tooth morphogenesis. The present study was performed to identify the genetic profiles which are involved in the morphological changes during the different stages of rat tooth development using the Agilent Rat Oligonucleotide Microarrays. Morphologically, the maxillary 3rd molar germ at 10 days post-partum (dpp) was at the cap/bell stage. In contrast, the maxillary 2nd molar germ showed the root development stage. After microarray analysis, there were a considerable number of up- or down-regulated genes in the 3rd and the 2nd molar germ cells during tooth morphogenesis. Several differentially expressed genes for nerve supply were further studied. Among them, neuroligin 1 (Nlgn 1) was gradually downregulated during tooth development both at the transcription and the translation level. Also, Nlgn 1 was mostly localized in the dental sac, which is an important component yielding the nerve supply. This genetic profiling study proposed that many genes may be implicated in the biological processes for the dental hard tissue formation and, furthermore, may allow the identification of the key genes involved in the nerve supply to the dental sac.

Vitellogenins (Vgs) are precursors of the major egg storage protein, vitellin (Vn), in many oviparous animals. Insects Vgs are large molecules (200-kD) synthesized in the fat body in a process that involves substantial structural modifications (e.g., glycosylation, lipidation, phosphorylation, and proteolytic cleavage, etc.) of the nascent protein prior to its secretion and transport to the ovaries. However, the extent to which Vgs are processed in the fat body varies greatly among different insect groups. We were cloned Vgs partial genes PaVgs and BgVgs from Periplaneta americana and Blattella germanica. Real-time quantitative PCR shows that PaVgs and BgVgs were differential-regulated with aging. In insects, glutathione S-transferases (GSTs) are enzymes involved in detoxification of insecticides. We were cloned GST partial genes PaGST and BgGST from Periplaneta americana and Blattella germanica. Real-time quantitative PCR shows that PaGST and BgGST were up-regulated with aging, and the mRNA level of PaGST and BgGST was higher in 4℃ and 37℃ than room temperature. The expression level of PaGST and BgGST exposure to temperature stress suggests that PaGST and BgGST are up-regulated after exposure low and hige temperature treatments.

Flammulina velutipes is a popular edible basidiomycete mushroom found in East Asia and is commonly known as winter mushroom. Mushroom development showing dramatic morphological changes by different environmental factors is scientifically and commercially interesting. We have sequenced the 35.6-megabase pair genome of F. velutipes KACC42780 using a next generation sequencing (NGS) approach. Then, HiSeq2000 sequencing system was used for comparing gene expression of F. velutipes genome at three different developmental stages (mycelium, primordium, and fruit body). Ninety one percents (10,116 genes) of the 10,999 predicted genes are expressed in at least one developmental stage. In addition, analysis of expressed genes in three different developmental stages showed that only 0.25–1.5% (28–165) of the total expressed genes (10,013) were uniquely expressed in a single developmental stage, whereas 83.6% (9,198) was shared in all stages.

In order to investigate genetic stability and gene expression profile after cloning procedure, two groups of cloned pigs were used for swine leukocyte antigen (SLA) gene nucleotide alteration and microarray analyses. Each group was consist of cloned pigs derived from same cell line (n=3 and 4, respectively). Six SLA loci were analyzed for cDNA sequences and protein translations. In total, 16 SLA alleles were identified and there were no evidence of SLA nucleotide alteration. All SLA sequences and protein translations were identical among the each pig in the same group. On the other hand, microarray assay was performed for profiling gene expression of the cloned pigs. In total, 43,603 genes were analyzed and 2,150～4,300 reliably hybridized spots on the each chip were selected for further analysis. Even though the cloned pigs in the same group had identical genetic background, 18.6～47.3% of analyzed genes were differentially expressed in between each cloned pigs. Furthermore, on gene clustering analysis, some cloned pigs showed abnormal physiological phenotypes such as inflammation, cancer or cardiomyopathy. We assumed that individual environmental adaption, sociality and rank in the pen might have induced these different phenotypes. In conclusion, the results of the present study indicate that SLA locus genes appear to be stable following SCNT. However, gene expressions and phenotypes between cloned pigs derived from the same cell line were not identical even under the same rearing conditions.

Osteoarthritis is one of the commonest causes associated with age-related damage of articular cartilage. Non-steroidal anti-inflammatory drugs are commonly used in osteoarthritic patient. However, long term administration of these drugs results gastrointestinal disorders. Though, most studies have demonstrated in the past that bee venom has therapeutic effect on diseases related to inflammation and pains, but its anti-inflammatory properties have not been so far studied on inflamed chondrocytes (LPS induced) invitro. For the purpose, the study was carried out to determine the effect of bee venom on porcine articular chondrocyte cell using microarray. In this study, we found that 2,235 significantly associated gene (1,404 up-regulated genes and 831 down-regulated genes) that were expressed on inflamed and non inflamed chondrocytes during proliferation. Among the 1,404 up-regulated genes and 831 down-regulated genes, known genes were 372 and 237, respectively. On the other hand, bee venom significantly reduced expression of fetuin involved in acute inflammatory reaction. Our results suggest that this study could be useful database in gene expression profiling of chondrocyte cell treated with bee venom.

To examine the expression profile of oxidative stress responsive genes in Spodoptera litura, we constructed a cDNA library from S. litura injected with hydrogen peroxide (H2O2). Using a microarray chip composed of 2,964 cDNAs, we screened gene expression at 1, 3, 5, 7, and 9 h post H2O2 injection. Data were clustered into 15 groups of genes that behave similarly across each time course. Seventy-three genes were identified as being at least 2-fold up- or downregulated after treatment with H2O2 in S. litura. We constructed expressed sequence tags (ESTs) for genes that changed at least 2-fold after treatment with H2O2. The functional classification of these ESTs based on Gene Ontology showed that the ESTs are rich in genes involved in oxidoreductase activity (5.7%), defense (14.3%), cellular process (22.9%), and development (17.1%).

Considering the great potential of iron chelators at inhibiting the proliferation of tumor cells, in order to determine the molecular and biological basis for the effects of iron chelator in oral cancer, we investigated the effects of iron chelator, desferrioxamine (DFO), on the gene profiling analysis of immortalized human oral keratinocytes (IHOK), and oral cancer cells (HN12), using the cDNA microarray. We identified 46 clones cDNA exhibiting more than 2 fold overexpression in DFO treated IHOK and HN12 cells, and 94 cDNA reveal more than 2 fold down-regulated expression. Examination of gene expression that differs between DFO treated vs. control IHOK and HN12 cells apprear to be related to : cell cycle regulator, cell growth and apoptosis, signal transduction and stress. p21 for cell cell cycle factor was upregualted, and cyclin-cdk gene was decreased expression, so we observed cell cycle arrest in DFO treated IHOK and HN12 cells. In tumor growth, we have identified downregulation of hemidesmosomal protein (bullous pemphigoid antigen 1) and epiregulin expression in DFO treated IHOK and oral cancer cells. Signal transducers including mitogen-activated protein kinase-activated protein kinase 5, serine/thereonine kinase 6 were downregulated with DFO treated cells, suggesting the DFO regulates the p38 MAP kianse pathway in immortalized and maignant oral keratincytes. In conclusion, we have demonstrated the high-throughput utility of cDNA array hybridization in parallel to the gene expression analysis to identify genes that are expressed differentially in DFO treated with immortalized and malignant oral keratinocytes. The differentially expressed genes identified here should be informative in DFO-induced anti-cancer effects.

To investigate the differential expression of genes by 635nm LEDs irradiation in arachidonic acid-treated human gingival fibroblasts, cDNA microarray was carried out. Human gingival fibroblasts were primary cultured and arachidonic acid was treated to induce inflammation. 635nm of wave length was used for LEDs irradiation. The experimental group was categorized into four group ; control, only LEDs irradiation group, only arachidonic acid-treated group and arachidonic acid-treated with LEDs irradiation group. The expression of 8,078 genes were increased and the expression of 7,103 genes were decreased in only LEDs irradiation group. For arachidonic acid-treated with LEDs irradiation group, the expression of 6,815 genes were increased, while the expression of 8,031 genes were decreased comparing with only arachidonic acid-treated group. IL-13alpha2 receptor was the most expressed gene in LEDs irradiation group comparing with control, followed by MMP3. Genes which the most down regulated was BIRC3 in LEDs irradiation group. PLAB genes was the most up-regulated in arachidonic acid treated with LEDs irradation group, followed by ranked RARRES1. Considering the classification by cell function, genes associated with signal transduction were the most affected by LEDs irradiation, followed by the genes associated with nucleoside, nucleotide and nucleic acid metabolism. In arachidonic acid treated with LEDs irradiation, genes associated with signal transduction and protein metabolism were affected. Taken together, LEDs irradiation could affect various biological process and could identify many genes related to LEDs irradiation, which could be used for clinical application.

A novel indirubin analog, 5'-nitro-indirubinoxime inhibits cell proliferation and induces apoptosis against various human cancer cells. In this study, we performed the microarray analysis to identify genes differentially expressed in the KB oral squamous carcinoma cells after treated with 5'-nitro-indirubinoxime. Among the 10,800 genes analyzed, 1,701 genes (15.8%) showed statistically different expression level in the 5'-nitro-indirubinoxime treated cells with respect to untreated control cells. Among those, 263 genes (15.5%) were down-regulated and 220 genes (12.9%) were up-regulated more than 2-fold. Functionally related gene clusters include genes associated with signal transduction (18.1%), especially genes related with apoptosis (3.5%) and cell cycle regulation (5.8%). Our application of microarray analysis on 5'-nitro-indirubinoxime treated oral cancer cells allows the identification of candidate genes for providing novel insights into the indirubin mediated antitumor activity.

A novel indil‘ubin analog‘ 5’ nitro-indirubinoxime(Ol1) inhibits cell proliferation and induces apoptosis again st variolls hllman cancer cell s. ln this stlldy, we performed the microarray analysis to identify genes diffel'enti ally expressed in the KB oral sqllamollS carcinoma cells after treatment with 011 Of the 10‘ 800 genes a nalyzed , 1700 genes(15.7%) showed di fferent expression level in the 011-treated cells with respect to untreated control cel1s Arnong those‘ 263 genes(15, 5%) were down -reg띠 ated and 220 genes(12, 9%) were IIp-regulated more than 2-fold, Functionally related gene clllsters inclllde genes associated with signal transdllction(18, 1%) , especially genes re lated with a poptosis(3, 5%) and cell cycle reglllation(5. 8%) . Our application of microarray ar뻐ysis on 01l-treated 01'외 cancer cells al lows the identifi cati on of candidate genes for providing novel insights into the 011-mediated anti -tllmor actl Vl ty ,

To identify genes implicated in the control of pluripotency as well as characteristics of stem cells, we analyzed expression profiles of genes derived from mouse morulas, blastocysts, embryonic stem cells, mesenchymal stem cells, and uterus tissue using cDNA microarray. Comparative analyses of their expression profiles identified putative clones that expressed specifically in specific samples or not in a specific sample. The expression pattern of these condidate clones was analyzed using RT-PCR and non-radioactive in situ hybridization. Functional annotation of these clones on pluripotency and stem cell plasticity is in ongoing. These studies may further our understanding on the nature of the stem cells and molecular mechanisms underlying many facets of mammalian development and differentiation.

Efficient infiltration of water through cell membranes is arbitrated by a family of transmembrane water channels called aquaporins (AQPs). Aquaporin belongs to a highly conserved group of membrane proteins called major intrinsic proteins that facilitate the transport of water and a variety of low molecular weight solutes across biological membranes,which is essential for plants to survive in stress conditions. This study identified 59 BrAQP genes from B. rapa database and Br135K microarray dataset, which was formed by applying low-temperature stresses to contrasting Chinese cabbage two inbreed lines, Chiifu and Kenshin. Based on phylogenetic analyses of BrAQPs revealed four distinct subfamilies, such as plasma membrane intrinsic proteins (PIP), tonoplast intrinsic proteins (TIP), NOD26-like intrinsic proteins (NIP), small basic intrinsic proteins (SIP) with aquaporin of Tomato and Arabidopsis thaliana. All BrAQP genes were firstly examined through homology study with existing biotic and abiotic stress resistance-related aquaporin genes of other plant species and found a high degree of homology. We selected PIP subfamily genes for expression analysis based on microarray data with high and differential transcript abundance levels and homology study with stress related aquaporin genes of other plant species. In our study, we characterized all B. rapa aquaporin genes and understanding the BrPIP subfamily gene function in plants under various environmental stimuli, the expressions of BrPIP genes under various abiotic stress conditions including cold, drought, salinity, water logging, ABA treatment and Fusarium oxysporum f. sp. Conglutinans infection were investigated by a quantitative real-time reverse transcription-PCR analysis. In our expression analysis, 4 BrPIP genes showed responsive expression against F. oxysporum f. sp. Conglutinans infection. The selected genes showed an organ-specific expression, and 12 out of 22 BrPIP genes were differentially expressed in Chiifu compared to Kenshin under cold stresses. Only 7 genes showed up regulation under drought stress and incase of salt stress 17 BrPIP genes were more responsiveness. Additionally, 18 BrPIP genes were up regulated by ABA treatment and all BrPIP genes showed down regulation under water logging stress. Together with expression and bioinformatic analyses, our results provides novel basis to allocate the stress-related biological function to each PIP gene.