Project description:cell culture:The human glioma cell line U87MG was obtained from the Cell Resource Center, Peking Union Medical College (Beijing, China), and U251MG was acquired from the American Type Culture Collection (Manassas, VA). Temozolomide (TMZ) resistant U87MG cells (U87TR) and TMZ resistant U251MG cells (U251TR) of glioblastoma (GBM) sub-cell lines, were established through repetitive exposure to increasing TMZ concentrations in vitro in our laboratory. Cells were cultured in DMEM culture medium supplemented with 10% FBS with a standard humidified incubator under 5% CO2 at 37°C.
Project description:Human hippocampus is one of the critical brain regions affected by aging. However, a fully understanding of aging-related changes in protein profiles in human hippocampus has not been well described, especially for the Chinese population. With the newly founded human brain bank in Chinese Academy of Medical Sciences & Peking Union Medical College, we performed a 4-plex TMT labeled proteomic study in the hippocampus of postmortem human brains. The ages of death ranged from 22-98, and were grouped into 4 aging groups: 20-50, 50-70, 70-90, and over 90 (n=4 each). None of the donors was diagnosed with neurodegenerative diseases according to their medical history. Our data identified 4582 proteins, among which 99 proteins were upregulated and 42 proteins were downregulated during the aging process.
Project description:Deep sequencing as a high-throughput technology has been widely used in the transcriptome profiling in mammals. In the present study, we aimed to identify chicken lncRNAs ranging from 300-1600 nt long. Total RNAs from chicken skeletal muscle at the embryonic stage were fractionated by 6% urea PAGE. Selected RNA fractions (300-1600 nt) were sequenced by Solexa technology. Chicken eggs (White Leghorn) were incubated at 37.5°C for 10 days (E10), 12 days (E12), 14 days (E14), 18 days (E18). Skeletal muscles (pectoralis) were collected from chicken embryo incubated at above four stages. All muscle samples were immediately frozen in liquid nitrogen and stored at -80°C until RNA isolation. All embryonic manipulations were conducted in accordance with the protocols of the Chinese Academy of Medical Sciences and the Institutional Animal Care and Use Committee of Peking Union Medical College. The single 'Chicken_muscle' Sample represents muscle pooled from all four stages.
Project description:The temporal lobe is the cerebral cortex with critical function. The superior and middle gyrus of temporal lobe have been well studied, however, present perceptions on inferior temporal gyrus remains limited. The understanding of age-related protein profile change in human inferior temporal gyrus has not yet been well established. This 3-plex TMT labeled proteomic study is performed based on the human brain bank at the Chinese Academy of Medical Sciences & Peking Union Medical College. Age distribution of the donors ranges from 22 to 90 years old, and were assigned to three age groups: 20-50, 50-70, and 70-90 years of death age. In this ageing cohort, no neurodegenerative disorders or major stroke events were identified via standard neuropathological classification. Proteomics and bioinformatics strategies were applied to identify the perturbations of protein expression and associated pathways. Among all the ITG samples, 3113 proteins were isolated, with 37 proteins upregulated and 21 proteins downregulated.
Project description:Human hippocampus is one of the critical brain regions affected by aging. However, a fully understanding of aging-related changes in protein profiles in human hippocampus has not been well described, especially for the Chinese population. With the newly founded human brain bank in Chinese Academy of Medical Sciences & Peking Union Medical College, we performed a 4-plex TMT labeled proteomic study in the hippocampus of postmortem human brains. The ages of death ranged from 22-98, and were grouped into 4 aging groups: 20-50, 50-70, 70-90, and over 90 (n=4 each). None of the donors was diagnosed with neurodegenerative diseases according to their medical history. Our data identified 4582 proteins, among which 99 proteins were upregulated and 42 proteins were downregulated during the aging process.
Project description:Most patients with papillary thyroid microcarcinoma (PTMC) have excellent prognoses with an indolent clinical courses. However, patients with lateral lymph node metastasis (LLNM) may experience higher locoregional recurrence rates and poorer prognoses. Therefore, it is crucial to establish an effective preoperative stratification strategy to reliably assess the risk of LLNM. PTMC samples were obtained from Peking Union Medical College Hospital. The data-independent acquisition mass spectrometry proteomics technique was used to identify protein profiles on PTMC tissues with and without LLNM.
Project description:Calcified aortic valve leaflets (CAVs) were explanted from patients with severe aortic valve stenosis undergoing aortic valve replacement at the Department of Cardiovascular Surgery, Union Hospital, affiliated to Tongji Medical College. Control non-calcified aortic valves with normal echocardiographic analyses were obtained during heart transplant procedures. RNA was extracted from valve leaflets and gene expression evaluated using the Arraystar Human mRNA Array. This study aimed to perform the expression analysis of mRNA on human aortic valves.
Project description:Transcriptional profiles of strains OG1RF (wild-type) and OG1RFΔmafR (deletion mutant). The goal was to determine the effect of the enterococcal MafR protein on global gene expression
Project description:The success of Enterococcus faecium and E. faecalis evolving as multi-resistant nosocomial pathogens is associated with their ability to acquire and share adaptive traits, including mobile genetic elements (MGE) encoding antimicrobial resistance. Here, we define the mobilome in representative successful hospital associated genetic lineages, E. faecium ST17 (n=10) and ST78 (n=10), E. faecalis ST6 (n=10) and ST40 (n=10) using DNA microarray analyses. The hybridization patterns of 272 targets representing plasmid backbones (n=85), transposable elements (n=85), resistance determinants (n=67), prophages (n=29), and CRISPR-cas sequences (n=6) separated the strains according to species, and for E. faecalis also according to STs. Although plasmids belonging to the RCR-, Rep_3-, RepA_N- and Inc18-families were well represented with no significant differences in prevalence, the presence of specific replicon classes differed highly between the species; E. faecium was dominated by rep17/pRUM, rep2/pRE25, rep14/EFNP1 and rep20/pLG1 and E. faecalis by rep9/pCF10, rep2/pRE25 and rep7. Tn916-elements conferring tetracycline resistance (tetM) were found in all E. faecalis strains, but only in two E. faecium strains. A significant higher prevalence of IS256-, IS3-, ISL3-, IS200/IS605-, IS110-, IS982-, and IS4-transposases were detected in E. faecium, and of IS110-, IS982- and IS1182-transposases in E. faecalis ST6 compared to ST40. Notably, the transposases of IS981, ISEfm1 and IS1678 which have only been reported in few enterococcal isolates, were well represented in the E. faecium strains. E. faecalis ST40 strains harboured possible functional CRISPR-Cas systems, and still resistance and prophage sequences were generally well represented. Gene targets defined as the enterococcal mobilome, including plasmids, IS elements and transposons, resistance determinants, prophage sequences and CRISPR-Cas systems were highly prevalent, underlining their potential importance in the evolution of hospital associated STs. An association between axe-txe to the RepA_N-family and ω-ε-ζ to the Inc18-family, implicates the contribution of TA-systems in stable plasmid maintenance carrying virulence and resistance determinants in enterococci. The concurrent presence of defined MGE and their associated resistance markers was generally confirmed and illustrates the importance of horizontal gene transfer in the development of multidrug resistant enterococci.