Project description:Titanium is a common implant material. However, in some patients titanium implants fail. Macrophages are key cells involved in foreign body response. To identify macrophage response to titainum, primary human macrophages were cultured on porous titanium discs for 6 days We used microarrays to determine the global expression pattern induced by porous titanium in macrophages and identify potential genes involved in implant failure.
Project description:Dry eye disease (DED) characterizes by chronic inflammation and an unstable tear film. Stem cells have shown potential for DED treatment, but the main challenge lies in improving the effectiveness of cell delivery. Here, we developed novel eye drops for DED treatment by employing porous microcarriers with mesenchymal stem cells. The microcarriers were created by electrospraying the solution of Arginine-Glycine-Aspartic Acid (RGD) peptides-modified sodium alginate with polyethylene oxide and mesenchymal stem/stromal cells (MSCs) into the calcium chloride solution. In vitro experiments based on a hyperosmotic corneal epithelial cell model indicated that porous microcarriers encapsulated with MSCs (RGD-Alg@MSCs) demonstrated notable enhancements in cell viability, reductions in apoptosis and reactive oxygen species (ROS), and diminished expression of pro-inflammatory cytokines. In the DED model using non-obese diabetic (NOD) mice, RGD-Alg@MSCs effectively enhanced tear production, promoted corneal healing, and alleviated inflammation. Additionally, RGD-Alg@MSCs modulated the immune environment in DED by inhibiting dendritic cell (DC) activation and suppressing Th17 differentiation in vitro, effectively disrupting the inflammatory feedback loop characteristic of DED. This immune modulation strategy was further validated through in vivo experiments, confirming its therapeutic potential. Thus, the designed MSCs-encapsulated porous microcarrier system can improve stem cell delivery and DED treatment efficiency.
Project description:The compaction of chromatin is a prevalent paradigm in gene repression. Chromatin compaction is commonly thought to repress transcription by restricting chromatin accessibility. However, the spatial organisation and dynamics of chromatin compacted by gene-repressing factors are unknown. Using cryo-electron tomography, we solved the three-dimensional structure of chromatin condensed by the Polycomb Repressive Complex 1 (PRC1) in a complex with CBX8. Unexpectedly, PRC1-condensed chromatin is porous and forms a size-selective diffusion barrier that is stabilised through multivalent dynamic interactions. Accordingly, PRC1 remains dynamic within the condensate while maintaining the chromatin static. In differentiated mouse embryonic stem cells, CBX8-bound chromatin remains accessible. These findings challenge the idea of open and closed chromatin states and instead suggest that PRC1 permits size-selective accessibility of macromolecules into chromatin condensates.
Project description:The compaction of chromatin is a prevalent paradigm in gene repression. Chromatin compaction is commonly thought to repress transcription by restricting chromatin accessibility. However, the spatial organisation and dynamics of chromatin compacted by gene-repressing factors are unknown. Using cryo-electron tomography, we solved the three-dimensional structure of chromatin condensed by the Polycomb Repressive Complex 1 (PRC1) in a complex with CBX8. Unexpectedly, PRC1-condensed chromatin is porous and forms a size-selective diffusion barrier that is stabilised through multivalent dynamic interactions. Accordingly, PRC1 remains dynamic within the condensate while maintaining the chromatin static. In differentiated mouse embryonic stem cells, CBX8-bound chromatin remains accessible. These findings challenge the idea of open and closed chromatin states and instead suggest that PRC1 permits size-selective accessibility of macromolecules into chromatin condensates.
Project description:The compaction of chromatin is a prevalent paradigm in gene repression. Chromatin compaction is commonly thought to repress transcription by restricting chromatin accessibility. However, the spatial organisation and dynamics of chromatin compacted by gene-repressing factors are unknown. Using cryo-electron tomography, we solved the three-dimensional structure of chromatin condensed by the Polycomb Repressive Complex 1 (PRC1) in a complex with CBX8. Unexpectedly, PRC1-condensed chromatin is porous and forms a size-selective diffusion barrier that is stabilised through multivalent dynamic interactions. Accordingly, PRC1 remains dynamic within the condensate while maintaining the chromatin static. In differentiated mouse embryonic stem cells, CBX8-bound chromatin remains accessible. These findings challenge the idea of open and closed chromatin states and instead suggest that PRC1 permits size-selective accessibility of macromolecules into chromatin condensates.
Project description:Mass spectrometry-based discovery glycoproteomics is highly dependent on the use of chromatography paradigms amenable to analyte retention separation. When compared against established stationary phases such as reversed phase and hydrophilic interaction liquid chromatography, reports utilizing porous graphitic carbon (PGC) have detailed its numerous advantages. Recent efforts have detailed the utility in porous graphitic carbon in high throughput glycoproteomics, principally through enhanced profiling depth and liquid phase resolution at higher column temperatures. However, increasing column temperature was shown to impart disparaging effects in glycopeptide identification. Herein we further elucidate this trend, describing qualitative and quantitative effects of increased column temperature on glycopeptide identification rates, signal intensity, resolution, and spectral count linear response. Through analysis of enriched bovine and human glycopeptides, species with high mannose and sialylated glycans were shown to most significantly benefit and suffer from high column temperatures, respectively. These results provide insight as to how porous graphitic carbon separations may be appropriately leveraged for glycopeptide identification while raising concerns over quantitative and pseudo-quantitative label free comparisons as temperature changes.
Project description:Purpose: The goal of this study is to compare endothelial small RNA transcriptome to identify the target of OASL under basal or stimulated conditions by utilizing miRNA-seq. Methods: Endothelial miRNA profilies of siCTL or siOASL transfected HUVECs were generated by illumina sequencing method, in duplicate. After sequencing, the raw sequence reads are filtered based on quality. The adapter sequences are also trimmed off the raw sequence reads. rRNA removed reads are sequentially aligned to reference genome (GRCh38) and miRNA prediction is performed by miRDeep2. Results: We identified known miRNA in species (miRDeep2) in the HUVECs transfected with siCTL or siOASL. The expression profile of mature miRNA is used to analyze differentially expressed miRNA(DE miRNA). Conclusions: Our study represents the first analysis of endothelial miRNA profiles affected by OASL knockdown with biologic replicates.
Project description:A cDNA library was constructed by Novogene (CA, USA) using a Small RNA Sample Pre Kit, and Illumina sequencing was conducted according to company workflow, using 20 million reads. Raw data were filtered for quality as determined by reads with a quality score > 5, reads containing N < 10%, no 5' primer contaminants, and reads with a 3' primer and insert tag. The 3' primer sequence was trimmed and reads with a poly A/T/G/C were removed