Project description:Primary uncultured CD31+ and CD105+ tumor endothelial cells (TEC), non-tumor endothelial cells (NEC) ,remnant cells from tumor (TC) and non-tumor liver tissue (NTC)of HCC tissues from patientswere isolated by magnetic-activated cell sorting. Affymetrix Human Gene ST Arrays were used to determine gene expression profiles of HCC cells and matched TEC and NEC cells. We use the gene expression profiling of the Human Genome U133 Plus 2.0 Arrays (Affymetrix, Santa Clara, CA) to identify distinct genes of TEC and NEC. Patient specimens were collected immediately after removal from patients with pathologically proven HCC and positive alpha-fetoprotein (AFP), then washed 3 times by phosphate buffer solution (PBS). Single cell suspension was obtained from separated tumor and non-tumor liver tissues of surgical specimens following the protocol (http://www.miltenyibiotec.com), then CD31+ cells or CD105+ cells were isolated using CD31 antibody or CD105 antibody conjugated magnetic beads in autoMACSpro (Meltenyi biotec).After the sample preparation, the gene expression profiling was performed using the Human Genome U133 Plus 2.0 Arrays (Affymetrix, Santa Clara, CA) according to manufacturer’s protocols at ShanghaiBio Corporation (Shanghai, China).
Project description:Hepatic stellate cells (HSCs) experience phenotypic transformation, from the quiescent phenotype to the activated one, after different etiologies of liver injury. Liver fibrosis is then occurred upon the activation of HSCs. miR-16 deficiency is identified to be an important characteristic of HSCs activation. We used Affymetrix rat 230 2.0 arrays (Affymetrix, Santa Clara, U.S.A.) to uncover the global alternations of transcriptome under miR-16 restoration. We isolated quiescent hepatic stellate cells (HSCs) from adult male SD rats (normal control group) by in situ perfusion and density-gradient centrifugation. Activated HSCs were separated from rats of fibrosis model group, which were treated by 40% carbon tetrachloride (CCl4) for 8 weeks, by means of liver section, digestion and sequential centrifugation. Quiescent and activated HSCs were then divided into 4 groups at random, namely quiescent HSCs, activated HSCs, pLV-miR-16-treated HSCs and pLV-GFP-treated HSCs. The pLV-miR-16-treated group, pLV-GFP-treated group were infected with pLV-miR-16 and pLV-GFP, respectively.
Project description:When PDMSCs were induced to heptocytes in vitro, cells mophology, stem cell markers, mitochondrial metabolism will change according to the differentiated status.But dedifferentiation reverses differentiated cells to a more primitive phenotype and PDMSCs will retain the multilineage potency. Furthermore, it will leads to the alteration of gene expression pattern. We used microarrays to detail the global programme of gene expression underlying dedifferentiation and hepatogenic differentiation prcocesses, we intend to identify distinct classes of differentiated genes during these processes. Human PDMSCs at passage 5 were induced to hepatocytes for 11 days, then the inductive medium was replaced by general culture medium for 1 day. Then human PDMSCs, hepatogenic PDMSCs at 11 days, dedifferentiated PDMSCs were selected for RNA extraction and hybridization on Affymetrix microarrays. To that end, we hand-selected cells at three time-points: before hepatogenic induction (P), hepatogenic PDMSCs at 11 days (H) and dedifferentiated PDMSCs for 1 day (DH) .
Project description:Compare difference Global expression profile of hiPSCs between hESCs and human Somatic cells, showing that hiPSCs and hESCs is consistent in lineages and indicated that the induce method is safe and reliable. There are three groups of samples, each group has two repeated samples, hiPSCs respectively compared with hESCs and human Urine-Derived Cells.
Project description:Long noncoding RNAs (lncRNAs) have been implicated in the formation of many different types of tumors. However, expression profiles and potential functions of lncRNAs in non-functioning pituitary adenomas (NFPAs) have not been systematically evaluated. We evaluated the expression profiles and potential functions of lncRNAs in non-functioning pituitary adenomas (NFPAs). 10 formalin-fixed and paraffin-embedded (FFPE) tissue specimens (5 non-functioning pituitary adenomas (NFPAs) and 5 normal pituitaries(NPs)) were selected for RNA extraction and hybridization on Affymetrix microarrays. The NFPAs team was designed as the Tumor group (T), while the NPs team was designed as Normal group (N).
Project description:The high concentration of Well5 cells was resuspended into 20μl PBS, the needle along the tibia direction, before reaching in a breakthrough sense, direct injection cells. At 7 days after injection, proximal tibia was able to reach mass production. At 20 days after injection, the proximal tibia mass increased.If prolonging exposure by BLI,this stage displayedthat tumor cell signalsbegan to lung metastasis. Osteosarcoma orthotopic lung metastasis model was successfully constructed. Total RNA was extracted from sorted osteosarcoma cells of the primary site and lung metastases using Trizol (Invitrogen). We used microarrays to detail the global programme of gene expression underlying cellularisation and identified distinct classes of up-regulated genes during osteosarcoma lung metastasis. In support of the notion that fibrosis marks the lung metastasis, the expression of numerous fibrosis-related genes such as FN1, COLs, and MMPs were upregulated from the primary site to lung metastasis in Well5-luc orthotopic inoculation model. Total RNA was extracted from sorted osteosarcoma cells using Trizol (Invitrogen). Gene expression profiling was conducted by Shanghai Biotechnology Corporation using Affymetrix U133 plus 2.0 arrays (Affymetrix, Santa Clara, CA). All data were analyzed according to the manufacturer’s protocol. Raw data generated from Affymetrix CEL files were normalized by RMA background correction; values were log2 transformed. For the enrichment of P values of each GO term, we used Fisher’s exact test to calculate P values and R package stats to calculate FDR (q value) by BH method (www.r-project.org).
Project description:MicroRNAs (miRNAs) are a class of endogenous non-coding RNAs of about 22 nt in length. Uveal melanoma is the most common intraocular malignant tumor in adults. Our previous result of microarray analysis showed that miR-142-3p was distinctly downregulated in uveal melanoma cells on which miR-142-3p was speculated to have important regulatory effect. In order to better understand the function of miR-142-3p in uveal melanoma and identify its gene targets, we performed transcriptomic microarray analysis. This was done by comparing gene expression profile changes in uveal melanoma cells transfected with miR-142-3p with that transfected with a negative control.
Project description:In rice (Oryza sativa L.), the number of panicles, spikelets per panicle and grain weight are important components of grain yield. These characteristics are controlled by quantitative trait loci (QTLs) and are derived from variation inherent in crops.The identification of different yield related QTLs facilitates an understanding of the mechanisms involved in cereal crop yield, and may have utility in improving grain yield in cereal crops. an understanding of the mechanisms involved in cereal crop yield, and may have utility in improving grain yield in cereal crops. In the present study, We cloned and characterized a large-panicle QTL, and confirmed that the newly identified gene OsEBS (enhancing biomass and spikelet number) increased plant height, leaf size and spikelet number per panicle, leading to an average of 37.62% increase in total grain yield per plant. trait loci (QTLs) and are derived from variation inherent in crops. OsEBS-transgenic rice B10201 and B10301 and control Guichao2
Project description:We knocked down EP300 and examined the expression of lncRNA625 target genes. Gene expression profiling of knockdown samples on cDNA microarrays indicated that EP300 affected expression of several lncRNA625 downstream target genes Stably-transfected KYSE150, transfected with shlncRNA625 or shscramble, were collected and lysed in TRIzol (Life technologies). Microarray experiments were performed following the Affymetrix protocol at the Shanghai Biotechnology Corporation.
Project description:To delineate the putative biological functions for lncRNA625, We performed expression profile from stably-transfected KYSE150 transfected with shlncRNA625 or shscramble for functions of lncRNA625 in ESCC Stably-transfected KYSE150, transfected with shlncRNA625 or shscramble, were collected and lysed in TRIzol (Life technologies). Microarray experiments were performed following the Affymetrix protocol at the Shanghai Biotechnology Corporation.