advanced method for isolation of mouse hepatocytes liver sinusoidal endothelial cells and kupffer cells
Methods in molecular biology vol 1540 hepatitis b virus methods and protocols
... Safiehkhatoon Moshkani, and Michael D Robek 21 Advanced Method for Isolation of Mouse Hepatocytes, Liver Sinusoidal Endothelial Cells, and Kupffer Cells Jia Liu, ... production in Huh-7 cells 12 Yinyan Sun et al Fig Southern blot analysis of cccDNA from HBV infected HepG2-NTCP cells HBV cccDNA is extracted from infected HepG2-NTCP (AC12) cells by Hirt method and ... viral mRNAs by sandwich hybridization Methods Enzymol 65(1):468–478 Chapter Hepatitis B Virus Infection of HepaRG Cells, HepaRG-hNTCP Cells, and Primary Human Hepatocytes Yi Ni and Stephan Urban...
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Báo cáo khoa học: E2A participates in a fine control of pre-mature B-cell apoptosis mediated by B-cell receptor signaling via transcriptional regulation of survivin, IAP2 and caspase-8 genes pot
... apoptosis in B-lymphocyte progenitor cells [19] However, the impact of B cell-specific factors, including E2A, on the BCR-mediated apoptosis of pre-mature B cells remains unclear, when apoptosis ... (PMA)/ionomycin and etoposide on 1420 cell-cycle progression and proliferation of DT40 and E2A)/) cells Cells cultured in the presence of PMA/ ionomycin for 24 h or etoposide for h were analyzed by ... symbols) 10 ngÆmL)1 PMA plus lM ionomycin at 37 °C for up to 48 h Viable cells were counted by the trypan blue dye exclusion method Data represent the mean of two separate experiments, and error...
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Báo cáo khoa học: Arg143 and Lys192 of the human mast cell chymase mediate the preference for acidic amino acids in position P2¢ of substrates pdf
... coding regions, the three different pCEP-Pu2 vector constructs were transfected into HEK 293 EBNA cells for protein production Recombinant protein was purified from conditioned media on Ni2+–nitrilotriacetic ... sequence has previously been cloned and inserted into the pCEP-Pu2 vector for expression in mammalian cells [4,23,24] This vector construct was used for in vitro mutagenesis of HC In this construct, ... lL of double-distilled H2O This DNA was then used to transform XL10-Gold ultracompetent E coli cells (provided by the manufacturer) All mutants were sequenced to confirm the inserted mutations...
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Báo cáo y học: "Specific eradication of HIV-1 from infected cultured cells" doc
... Specific killing of HIV-1 infected cells (a) H9 lymphocyte T cells were infected by the WT HIV-1 at MOI of 0.1, exactly as described in [28], and then the infected cells were treated every two days ... TZM-bl cells at 48 h PI (c) Same as (a) but cells viability was estimated by the MTT assay as described in [28] (dI) and (dII) Same as (a) but the average amount of viral cDNA integration events /cells ... cultured cells died by the eighth day PI following the addition of the Mix This percentage may represent the relative amount of virus infected cells, probably indicating total death of these cells...
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Báo cáo sinh học: "The combined transduction of copper, zincsuperoxide dismutase and catalase mediated by cell-penetrating peptide, PEP-1, to protect myocardium from ischemia-reperfusion injury" ppt
... SOD1 and CAT can not be delivered into living cells because of the poor permeability and selectivity of the cell membrane, which has limited its usage in protecting cells/ tissues from oxidative ... protein into H9c2 cells H9c2 cells were grown to confluence on 25 cm2 flasks and pretreated with PEP-1-SOD1-His or PEP-1-CATHis at different doses (0.5~2.0 μM) for 15 min~72 h The cells were then ... PEP-1-CAT fusion protein into H9c2 cells, cells were treated with μM of control SOD1, purified PEP-1SOD1, CAT, or PEP-1-CAT After or h of incubation at 37°C, the cells were washed twice with × PBS...
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Tài liệu Báo cáo khoa học: Properties of ecdysteroid receptors from diverse insect species in a heterologous cell culture system – a basis for screening novel insecticidal candidates docx
... aspects of cell culture methodology, ligand application, transfection, reporter gene measurement, western immunoblotting and EMSAs have been previously reported [9,21] Briefly, CHO cells were grown to ... USP-encoding vectors described below After transfection for h, cells were incubated with or without agonists and ⁄ or JHIII for 24 h, cells were harvested, and extracts were processed for the studies ... Drosophila EcR– USP heterodimer is potentiated by the presence of juvenile hormone (JH) in mammalian cells; that is, JH dramatically reduces the ecdysteroid concentration necessary to attain maximal...
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Báo cáo khoa học: Characterization of a second proliferating cell nuclear antigen (PCNA2) from Drosophila melanogaster docx
... of cellular proteins S2 cells were exposed to MMS or mitomycin C for h or to H2O2 for 15 The cells were then washed once and incubated prior to sampling UV-irradiated S2 cells were incubated in ... 0.02% mitomycin C (MMC)] S2 cells were collected at the indicated post-treatment intervals (D) Chromatin binding of DmPCNA2 after various doses of DNA-damaging agents S2 cells were treated with MMS ... anti-GST serum S2 cells were placed on poly-(l-lysine)-coated coverslips and fixed with 4% paraformaldehyde in NaCl ⁄ Pi for 10 at room temperature After several washes with NaCl ⁄ Pi, the cells were...
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báo cáo hóa học:" Regulatory activity of azabisphosphonate-capped dendrimers on human CD4+ T cell proliferation enhances ex-vivo expansion of NK cells from PBMCs for immunotherapy" potx
... support that T cells constitute a barrier for the expansion of NK cells IL-2 stimulation of PBMCs was shown to elicit absolute expansion of NK cells and CD56+ T cells, e.g NKT cells, γδ T cells and ... proportion of cells in the G1 or G2/M phase of mitosis versus cells undergoing nucleus fragmentation A very slight increase in the percentage of apoptotic cells was observed when cells were cultured ... regulatory T cells 3a-G1 prevents IL-2 driven expansion of CD4+/Foxp3high regulatory T cells a) Expanded CD4+/CD25+ T cells among IL-2 treated PBMCs present characteristics of regulatory T cells, e.g...
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báo cáo hóa học: " Quality of life among adolescents with sickle cell disease: mediation of pain by internalizing symptoms and parenting stress" pdf
... with SCD and disease-related parenting stress would mediate the association of pain with HRQOL Methods Participant Recruitment The current data were collected as part of a larger study examining ... 2006 by the study investigators and trained research assistants (teams of doctoral students and advanced undergraduates) The protocol was approved by the appropriate Institutional Review Board ... Press; 1995:105-123 Pal DK: Quality of life assessment in children: a review of conceptual and methodological issues in multidimensional health status measures J Epidemiol Community Health 1996,...
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Báo cáo khoa học " Structural characterization of immunostimulating polysaccharide from cultured mycelia of Cordyceps militaris " doc
... Carbohydrate Polymers 80 (2010) 1011–1017 Materials and methods 2.5 Determination of NO production 2.1 Materials After pre-incubation RAW264.7 cells (1 Â 106 cells/ ml) for 18 h, each polysaccharide (1000 ... reduction of a tetrazolium salt by mitochondrial dehydrogenase in viable cells After pre-incubating RAW264.7 cells (1 Â 106 cells/ ml) for 18 h, polysaccharides (1000 lg/ml) or LPS (2.5 lg/ml) was ... Bradford method (Bradford, 1976), the hexosamine content was evaluated using the Elson–Morgan method (Dische, 1962), and the uronic acid content was assessed using the Blumenkrantz method (Blumenkrantz...
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Báo cáo y học: " Monocytes are essential for inhibition of synovial T-cell glucocorticoid-mediated apoptosis in rheumatoid arthritis" docx
... analyzed by flow cytometry T cells were gated as CD3+ cells, and apoptosis was quantified as the mean percentage of Annexin V+ cells from the total number of gated cells Transwell coculture experiments ... analyzed by flow cytometry T cells were gated as CD3+ cells, and apoptosis was quantified as the mean percentage of Annexin V and 7AAD+ cells from the total number of gated cells Statistical analysis ... minimal changes in the inflammatory phenotype of the endothelial synovial cells [15] Our current results showing resistance of RA synovial T cells to glucocorticoid-induced apoptosis provide further...
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Báo cáo y học: "CD14 mediates the innate immune responses to arthritopathogenic peptidoglycan–polysaccharide complexes of Gram-positive bacterial cell walls" docx
... collected and washed thee times, and cells were allowed to adhere to plastic for 30 min, followed by removal of nonadherent cells Unless otherwise indicated, cells were maintained for 24 hours ... knockout cells At the saturating concentrations of PG-PS (20 µg/ml), the maximal increase in [Ca 2+]i in the wild-type cells was 47 ± nmol/l as compared with 12 ± nmol/l in CD14 knockout cells (Fig ... show the following: top – unstimulated cells from wild-type mice; middle – PG-PS stimulated cells from wild-type mice; and bottom row – PG-PS-stimulated cells from CD14 knockout mice Representative...
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báo cáo khoa học: "A Review of Duodenal Metastases from Squamous Cell Carcinoma of the Cervix Presenting as an Upper Gastrointestinal Bleed" potx
... nonkeratinizing squamous cells( *) similar to the cells seen in Figure 2a(*) 3b: Photomicrograph of staining with Cytokeratin shows strong cytoplasmic and membrane staining of the lesional cells (*) as seen ... nonkeratinizing squamous cells( *) similar to the cells seen in Figure 2a (*) 4b: Photomicrograph of staining with Cytokeratin shows strong cytoplasmic and membrane staining of the lesional cells (*) as ... mucosal layer including the absorptive, glandular, and neuroendocrine cells is restored every 4-7 days Such high turnaround of mucosal cells is probably incompatible with the “critical cell mass” required...
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Báo cáo khoa học: "In vitro characterization of cells derived from chordoma cell line U-CH1 following treatment with X-rays, heavy ions and chemotherapeutic drugs" pdf
... triangle indicates HeLa cells, white circle indicates U87-MG glioma cells, and black circle presents chordoma origin U-CH1-N cells B) Chromosome number of chordoma origin U-CH1-N cells 50 metaphase ... more than 200 cells (more than doublings), and the most of the colonies from irradiated cells contained more than 100 cells (more than doublings) for U-CH1-N Small colonies with 1020 cells (less ... time, cells were seeded at 5000 cells per T12.5 flask, and their number was counted at regular intervals Comparison of parental and subpopulation of U-CH1, chromosome and p53 analysis U-CH1-N cells...
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Báo cáo y học: "Monocytes are essential for inhibition of synovial T-cell glucocorticoid-mediated apoptosis in rheumatoid arthritis" pdf
... analyzed by flow cytometry T cells were gated as CD3+ cells, and apoptosis was quantified as the mean percentage of Annexin V+ cells from the total number of gated cells Transwell coculture experiments ... analyzed by flow cytometry T cells were gated as CD3+ cells, and apoptosis was quantified as the mean percentage of Annexin V and 7AAD+ cells from the total number of gated cells Statistical analysis ... minimal changes in the inflammatory phenotype of the endothelial synovial cells [15] Our current results showing resistance of RA synovial T cells to glucocorticoid-induced apoptosis provide further...
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báo cáo khoa học: "Enhancive effects of Lewis y antigen on CD44-mediated adhesion and spreading of human ovarian cancer cell line RMG-I" pps
... randomly selected to count 200 cells: the round and bright cells were counted as non-spreading cells; the oval cells with pseudopods were counted as spreading cells Irrelevant control antibodies ... cells detected by immunocytochemistry (×400) Panels and are negative controls; panels and are Lewis y antibody-untreated cells; panels and are Lewis y antibody-treated cells; panels and are cells ... RMG-I-H cells The 2-ΔΔCT value of mRNA level of CD44 in RMG-I-H cells is 79% of that in RMG-I cells, which had no significant difference (P > 0.05), whereas the mRNA level of a1,2-FT in RMG-I-H cells...
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báo cáo khoa học: " Effect of filtration on morphine and particle content of injections prepared from slow-release oral morphine tablets" ppt
... prepared injections from tablets and their filtration procedures The methods used in this laboratory study are based on these methods Extraction of morphine from tablets One sustained-release 60 ... users employ a variety of improvised methods, such as crushing the tablet between two spoons However, it was considered preferable to use a single, reproducible method in this study, although it ... would be lost in filtration The current study was designed to replicate preparation and filtration methods used by injecting drug users for injection of slow-release oral morphine tablets (MS Contin®,...
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Báo cáo y học: " Analysis of Prototype Foamy Virus particle-host cell interaction with autofluorescent retroviral particles" ppt
... imaging CLSM analysis of PFV binding to host cells Host cells were seeded at a density of 1.5 × 104 cells/ well into well chamber slides After 24 h cells were cooled down and incubated on ice with ... mock-treated cells (mock) (Fig 7B) However, a significant binding activity of Envdeficient PGM3-GFP particles (PGM3-GFP ΔEnv) was detected on HeLa cells in comparison to mock-incubated cells (mock), ... synthesis deficient Sog9 cells in comparison to the parental mouse L -cells (Fig 8C) This is in line with the transduction susceptibility data of these cell lines Interestingly, Pac2 cells still specifically...
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Báo cáo y học: "Adoptive transfer of splenocytes to study cell-mediated immune responses in hepatitis C infection using HCV transgenic mice" pptx
... lymphocytes in the liver C) Immunofluorescence analysis of frozen liver sections (5 μm thick) of non-transgenic mouse showing no CFSE labeled cells over the liver section D) H&E staining of liver sections ... ligand on the surface of liver- infiltrating T cells leading to the induction of hepatic inflammation and liver damage [12,13] Another important mechanism of immune-mediated liver damage is through ... (CFSE)-labeled T cells from HCV immunized mice homed to the liver of HCV transgenic mice, indicating that these HCV-activated T cells recognize the HCV transgene and attack the hepatocytes expressing...
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