Project description:The general aim of this paper is to explore a little further into molecular biology of the interaction between virus and host cell. We seek to use cDNA microarray technology, one revolutionary methodological advance, to perform a high throughput analysis of cDNA clones with the intent of identifying genes expressed in association with the infection with BKV in Vero cells (green monkey kidney cells). This technology, which can supply quantitative expression information for many thousands of genes simultaneously, has been used to classify the cellular genes at transcript levels in different physiological states of a tissue or cell. Accordingly, in this study, we have constructed a cDNA microarray containing 32448 spots (quadruplication of 7,334 human cDNAs and doping controls) to study that BKV infection of Vero cells. Our purposes were (i) to appraise whether cDNA microarrays could be employed to investigate the expression of cellular genes during the 5 time courses (5, 10, 15, 19 and 25 days post-infection (dpi)) of infection with BKV, (ii) to identify any genes that resulted in up-regulation or down-regulation of cell gene expression transcription, (iii) to determine the etiological role of BKV in nephropathy and/or neoplasia, (iv) to form a clearer picture of virus-associated pathophysiology in kidney. Keywords: BK virus, BKV, cDNA microarray, Vero cell We used a loop design in this study. cDNA microarray experiment consisted of ten RNA samples, including BKV-infected samples and their corresponding uninfected controls in 5 different time points. See supplementary PDF file for additional information
Project description:The general aim of this paper is to explore a little further into molecular biology of the interaction between virus and host cell. We seek to use cDNA microarray technology, one revolutionary methodological advance, to perform a high throughput analysis of cDNA clones with the intent of identifying genes expressed in association with the infection with BKV in Vero cells (green monkey kidney cells). This technology, which can supply quantitative expression information for many thousands of genes simultaneously, has been used to classify the cellular genes at transcript levels in different physiological states of a tissue or cell. Accordingly, in this study, we have constructed a cDNA microarray containing 32448 spots (quadruplication of 7,334 human cDNAs and doping controls) to study that BKV infection of Vero cells. Our purposes were (i) to appraise whether cDNA microarrays could be employed to investigate the expression of cellular genes during the 5 time courses (5, 10, 15, 19 and 25 days post-infection (dpi)) of infection with BKV, (ii) to identify any genes that resulted in up-regulation or down-regulation of cell gene expression transcription, (iii) to determine the etiological role of BKV in nephropathy and/or neoplasia, (iv) to form a clearer picture of virus-associated pathophysiology in kidney. Keywords: BK virus, BKV, cDNA microarray, Vero cell
Project description:Infectious bursal disease virus (IBDV) is the pathogenic agent of infectious bursal disease (IBD). Scine it was observed in 1957, IBD spread worldwidely in the chicken flocks, is a important immunosuppressive disease and an threat to poultry industry. Although many studies have be done about IBDV, interaction of IBDV infection and IBDV-encoding genes to host cell gene expression are little known. In this study, the LongSAGE library of Vero-cell, IBDV- infected vero cell, Vero-cell transfected with IBDV-VP5 gene, Vero-cell transfected with IBDV A frament and Vero-cell transfected with IBDV VP243 frament were obtained. We got 96,213 gene tags (17 nucleotides), which represented 24,475 transcripts. Keywords: Transcripts of different state vero-cell
Project description:To compare MicroRNA expression in Vero cells infected with DENV-2 adapted strain of Vero cells and its source srain derived from C6/36 cells
Project description:Infectious bursal disease virus (IBDV) is the pathogenic agent of infectious bursal disease (IBD). Scine it was observed in 1957, IBD spread worldwidely in the chicken flocks, is a important immunosuppressive disease and an threat to poultry industry. Although many studies have be done about IBDV, interaction of IBDV infection and IBDV-encoding genes to host cell gene expression are little known. In this study, the LongSAGE library of Vero-cell, IBDV- infected vero cell, Vero-cell transfected with IBDV-VP5 gene, Vero-cell transfected with IBDV A frament and Vero-cell transfected with IBDV VP243 frament were obtained. We got 96,213 gene tags (17 nucleotides), which represented 24,475 transcripts. Keywords: Transcripts of different state vero-cell 1.Cloning of the full-length genomic A-segment, VP5 ORF cDNA, VP243 ORF cDNA of IBDV 2.Establishing cloned Vero cell lines expressing VP5, VP243 and A fragment of IBDV 3.Construction of Long-SAGE libraries 4. Sequencing
Project description:Two rounds of TMT relative quantitative proteomics were performed to detect cellular factors involved in p-eIF4E regulation of the synthesis of viral proteins.our first round of screening identified differentially expressed proteins in PEDV-infected cells and mock-infected cells; the cellular pathways involved were mainly the estrogen, cAMP, and calcium signaling pathways. Second round screening identified differentially expressed proteins in the PEDV-infected S209A-Vero cells vs. the PEDV-infected WT-Vero cells; the regulated cellular pathways were found to be mainly in the PI3K-Akt, focal adhesion, and mTOR signaling pathways, and the biological processes and molecular functions in which p-eIF4E played a role were related mainly to metabolism and biogenesis, catalytic activity, and stimuli response.4006 host factors were detected, of which 193 (in brown) were significantly upregulated (ratio ≥1.2, P<0.05) and 191 (in green) were down-regulated upon PEDV infection (ratio ≤0.83, P<0.05). 29 of the 191 down-regulated proteins were susceptible to a low level of p-eIF4E . Notably, among the 193 upregulated cellular proteins, 77 were upregulated in the WT-Vero over the S209A-Vero cells , suggesting that the WT-Vero cells are more susceptible to a high level of p-eIF4E.
Project description:Intervertebral disc degeneration(IVDD) is a common spinal condition with limited effective treatments available. This study aims to investigate the impact of poly(lactic-co-glycolic acid)/Bradykinin (PLGA/BK) microspheres on IVDD and its underlying mechanisms. We collected nucleus pulposus samples from both healthy and degenerated human intervertebral discs and conducted immunohistochemical analyses, revealing reduced BK expression in degenerated tissues. Subsequently, we used BK to treat nucleus pulposus cells and conducted Bulk RNA sequencing (RNA-seq), identifying BK's involvement in cellular senescence, extracellular matrix metabolism, and the PI3K signaling pathway. Further experiments using tert-butyl hydroperoxide (TBHP)-induced cell senescence showed that BK treatment reduced senescence, enhanced extracellular matrix synthesis, and inhibited degradation, along with activation of the PI3K pathway. These effects were mediated through B2R (BK receptor 2) and the downstream PI3K pathway. Following this, we developed sustained-release BK microspheres with an optimized manufacturing process. In vitro co-culture experiments showed no observable toxicity. We established an IVDD model in rat tail vertebrae through fine needle puncture, administering local injections of BK sustained-release microspheres. Using various experimental methods, including X-ray, MRI, histopathology, and immunohistochemistry, we found that these microspheres could slow the progression of IVDD. This study highlights the potential of injectable PLGA/BK microspheres to regulate cellular senescence and extracellular matrix metabolism via the B2R and PI3K pathways, ultimately delaying IVDD.