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Objective To investigate the effects of 620nm noncoherent red light emitted from light-emitting diode (LED) on proliferation and osteogenic differentiation of rat osteoblasts.Methods Osteoblasts were isolated from SD rat and cultured in vitro.The LED was placed at 2cm above the cell layer.Cells were divided into four groups and each group was irradiated for 0s, 150s, 300s and 600s at energy dose of 0, 1, 2, and 4J/cm2, respectively.Cells were irradiated once every day.Cellular proliferation activities were evaluated by using cell counting kit-8 ( CCK-8 ) at 2nd and 4th d.The alkaline phosphatase (ALP) ratio activity was evaluated by alkaline phosphatase activity kit at the 9th d, and expressions of osteoblast master genes ( Collα1, Bglap and Runx2) were monitored as indicators of osteoblast differentiation by reverse transcription-polymerase chain reaction (RT-PCR) technique.Results In the group irradiated with 1 and 2 J/cm2 proliferation activities of osteoblasts enhanced significantly compared with the control group at the 4th d ( P < 0.05 ).In the group irradiated with 4 J/cm2 ALP ratio activity elevated significantly compared with the control group at the 9th d ( P <0.05 ).In the group of red light irradiation expressions of Bglap and Runx2 enhanced significantly.Conclusion The 620nm nonconherent red light emitted from LED can promote proliferation of osteoblasts and enhance osteogenic differentiation significantly.
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Objective To study the effects of an electromagnetic field (EMF) on the expression of fibro-blast growth factor (FGF-2) and it' s receptor (FGFR-2) mRNA in rat bone marrow mesenchymal stem cells (BMSCs) in vitro. Methods Rat BMSCs were isolated and cultured in vitro. The subcultured cells were divided into different groups to be EMF stimulated at 1.0 mT. The expression of FGF-2 and FGFR-2 mRNA were detected by reverse transcription-polymerase chain reaction (RT-PCR). Results Different frequencies and durations of 1.0 mT EMF exposure induced FGF-2 and FGFR-2 mRNA expression in comparison to blank controls. The expression of FGF-2 mRNA reached a peak after stimulation at 15 Hz for 10 min, 50 Hz for 60 min and 75 Hz for 30 min. And the expression of FGFR-2 mRNA reached a peak after 30 minutes at all frequencies. At 1.0 mT with 30 min exposure, the expression of FGF-2 mRNA peaked after 50 Hz stimulation, and the expression of FGFR-2 mRNA peaked after stimulation at 75 Hz. Conclusions Moderate EMF stimulation can significantly increase the expression of FGF-2 and FGFR-2 mRNA in rat BMSCs in vitro.
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Objective To study the effects of electromagnetic field (EMF) exposure on osteoporosis in ovariectomized mice. Methods Sixty 8-week-old female Kunming mice were divided into four groups at random: a sham operation group (group A), an ovariectomized group (group B), an EMF and ovariectomized group (group C) and a nilestriol and ovariectomized group (group D). Bilateral ovariectomies were performed on all mice except those in group A. The mice of group C were exposured to a 15 Hz, 1.0 mT electromagnetic field. The mice of group D were given at nilestriol 1.5 mg/kg/week. The bone mineral density (BMD) of the lumbar vertebrae was measured before the mice were sacrificed at the 12th week. Blood specimens were collected every two weeks to measure the ac-tivity of alkaline phosphatase (ALP) and the concentration of bone gamma-carboxyglutamic-acid-containing proteins (BGP), calcium and estradiol in the serum. Histological sections were taken to examine and analyze the changes in bone trabeculae in the lumbar vertebrae after 6 and 12 weeks. Results EMF at 15 Hz and 1.0 mT intensity signifi-cantly increased the activity of ALP and the concentrations of BGP and calcium in the serum. In addition, the absorp-tion of bone trabeculae in the lumbar vertebrae was significantly restrained. Conclusions EMF at 15 Hz and 1.0 mT can restrain the development of osteoporosis in ovariectomized mice.
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To obtain the recombinant 18 kD protein with high purity and normal bioactivity of Cysticercus cellulosae (rCE18), E. coli cells with the rCE18 were disrupted ultra-sonically, and the inclusion bodies were washed with a solution containing 0.2% deoxycholic acid sodium (DOC)and 2% DOC, respectively. Then they were denatured with 0.9% sodium lauroyl sarcosine (SKL) followed by dialysis and gel filtration to refold and purify the target protein. At the same time, this method was compared with GST-FF affinity chromatography and recovering from SDS-PAGE gel. Biological activity of purified rCE18 was analyzed with indirect ELISA, and the purity of the products was identified using SDS-PAGE. The purity of refolded inclusion bodies exceeded 60% and the total recovery of activated protein rCE18 was about 41.3%. The specificity of rCE18 reached up to 97.2% using indirect ELISA. An effective way for purifying and refolding rCE18 expressed in E. coli as inclusion bodies was established, rCE18 with higher purity and activity was obtained, which has the potential for developing diagnosis methods of porcine cysticercosis.
Subject(s)
Animals , Antigens, Helminth , Genetics , Allergy and Immunology , Chromatography, Gel , Cysticercus , Genetics , Allergy and Immunology , Metabolism , Escherichia coli , Genetics , Metabolism , Inclusion Bodies , Metabolism , Protein Renaturation , Recombinant Proteins , Genetics , Allergy and ImmunologyABSTRACT
The reasonable distribution of healthcare human resources is the fundamental requirement to improve medical efficiency and service quality,satisfy public's diverse requirement for medical care,and realize medical service fairness.However,various factors hinder the reasonable distribution of healthcare human resources and the realization of medical service fairness in the present medical system,including the imperfect system of human resources management,the immature system of performance appraisal,the imbalanced development of different regions,and the unpleasant environment of medical practice.This paper reflects on those issues from an ethical perspective.
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Objective To obtain related genes of Cysticercus cellulosae from spliced leader (SL) cDNA library. Methods Spliced leader library of Cysticercus cellulosae was constructed using SL specific primer and oligo (dT)15 with M13M4 primer, and positive clones were then screened randomly, identified with enzyme restriction, followed by sequencing and homologous analysis. Results The amino acid sequence, encoded by the positive clone with a poly (A) 22 tail and a complete open reading frame (ORF), was with homology of RNA polymerase subunit genes of human, B. napus, fission yeast, A. thaliana, C. elegans and fruit fly up to 71.6%. Conclusion The protein, RNA polymerase subunit encoded putatively by the clone, is high conservative in different species.