ABSTRACT
OBJECTIVE@#To observe the effect of Bruton's tyrosine kinase (BTK) on the proliferation and differentiation of osteoclasts and to explore the mechanism of BTK on bone destruction in periapical periodontitis.@*METHODS@#After RAW264.7 cells induced with 100 ng·L⁻¹ receptor activator for nuclear factor-κB ligand (RANKL) for 5 days, osteoclast induction was confirmed by light microscopy, tartrate-resistant acid phosphatase (TRAP) staining, and quantitative real-time PCR (RT-qPCR). Then, BTK-small interfering RNA (BTK-siRNA) was transfected into cells induced for 5 days. After 24 h, the expression of TRAP mRNA was measured using RT-qPCR, and the proliferation and differentiation of osteoclasts were detected using CCK-8 and TRAP activity assay. Statistical analysis was performed.@*RESULTS@#After RAW264.7 was induced with RANKL for 5 days, a large number of round, ellipse, irregularly protuberant, and TRAP-positive macrophages were observed under light microscopy. The expression of TRAP mRNA significantly reduced after 24 h of BTK-siRNA transfection (P<0.05). The detection of CCK-8 and TRAP activities showed that the proliferation and differentiation of osteoclasts significantly decreased (P<0.05).@*CONCLUSIONS@#Silencing of BTK can inhibit the proliferation and differentiation of osteoclasts. BTK can be used as a new target for the inhibition of osteoclasts.
Subject(s)
Agammaglobulinaemia Tyrosine Kinase , Cell Differentiation , Cell Proliferation , Macrophages , Osteoclasts , RANK LigandABSTRACT
@#Objective To summarize the experience of combined application of low and high frequency ultrasound in the diagnosis of acute primary epiploic appendagitis. Methods The clinical data of 130 patients with acute primary epiploic appendagitis confirmed by operation or other clinical examinations were retrospectively analyzed. Results All the 130 patients were found homogenous or heterogeneous solid block in the area of the abdominal pressure point. Four patients were treated by operation and 126 patients were cured by symptomatic supportive treatment. There were no serious complications in all patients. Conclusion Ultrasonography plays an important role in the diagnosis and differential diagnosis of acute primary epiploic appendagitis.
ABSTRACT
A quick HPLC-ESI-MS/MS method was established for simultaneous determination of four major diterpenoids in Rabdosia japonica var.glaucocalyx, including glaucocalyxin A, oridonin, hebeirubesensin and enmenol. Analysis was performed on an Agilent ZORBAX SB-C18(4.6 mm×250 mm, 5 μm ) column eluted in a gradient program with methanol and water. The flow rate was 0.8 mL•min⁻¹. Multiple reaction monitoring (MRM) scanning mode was performed in negative ion switching mode to apply for the quantitative determination. The calibration curves for the above four compounds were linear in corresponding injection amount. The average recoveries of the compounds ranged from 92.40% to 105.9%, with RSDs of 1.7%-6.5%. The method is simple, rapid, accurate with good repeatability, which can provide a reference for overcalling evaluation the quality of R. japonica var.glaucocalyx. The result of cluster analysis- showed that the quality of R. japonica glaucocalyx var. greatly varied between areas and parts.
ABSTRACT
A quick HPLC-ESI-MS/MS method was established for simultaneous determination of three chemical compositions in Usnea, including usnic acid, diffractaic acid, and ramalic acid. The separation was performed on a chromatographic column of Agilent ZORBAX SB-C, (4.6 mm x 250 mm, 5 µm), and the mobile phase was methanol (0.05% formic acid)-0.05% formic acid solution (4 mmol ammonium acetate), with an isocratic elution at a flow rate of 0.8 ml · min⁻¹. Multiple reaction monitoring scanning mode (MRM) was performed combined with the ion switching technology in positive and negative ion switching mode to apply for the quantitative determination. The calibration curves for the above three compounds were linear in corresponding injection amount. Their average recoveries were 95.0%-105.1%, with RSDs of 1.1%-5.2%. The method was simple, rapid, accurate with high repeatability, which could provide a reference for overcalling evaluation the quality of Usnea.
Subject(s)
Chromatography, High Pressure Liquid , Methods , Hydroxybenzoates , Spectrometry, Mass, Electrospray Ionization , Methods , Tandem Mass Spectrometry , Methods , Usnea , ChemistryABSTRACT
Random amplified microsatellite polymorphism (RAMP) markers were used to access the genetic diversity among 112 samples of nine populations of Dendrobium officinale Kimura et Migo. Using 16 informative primers, 123 bands were amplified and 86 (69.92%) were polymorphic. The polymorphic bands from three to eight could be detected for each RAMP primer, with a mean of 5, indicating abundant genetic diversity among populations. Genetic similarity coefficients ranged from 0.250 to 0.813. UPGMA dendrogram illustrated 9 populations clustered into 3 groups, and the cluster pattern showed correlation with the locations of the D. officinale populations. These results were supported by the previous conclusions that were achieved by other molecular markers, and RAMP is proved to be effective for evaluating the genetic diversity of wild populations of Dendrobium officinale.
Subject(s)
Cluster Analysis , DNA Primers , DNA, Plant , Genetics , Dendrobium , Genetics , Genetic Variation , Microsatellite Repeats , Phylogeny , Plants, Medicinal , Genetics , Polymerase Chain Reaction , Methods , Polymorphism, Genetic , Random Amplified Polymorphic DNA Technique , MethodsABSTRACT
Objective To investigate the effects of the HLA-Ⅰ and co-stimulators CD_(80)and CD_(86)on HT-29 cells surface, after heat killed Bifidobacterium and HT-29 cells were mingled culturing for 12 hours. Methods The HLA-Ⅰ and co-stimulators CD_(80)and CD_(86)on HT-29 cells surface were measured by Flow Cytometer. Results Heat killed Bifidobacterium can improve the expressions of HLA-1, CD_(80)and CD_(86)on HT-29 cells significantly, the livability of colon carcinoma cell was more than 90 %. Conclusion Heat killed Bifidobacterium have potential antitumorous features.