Objective To investigate the effect of genistein, a legume-derived isoflavone,on the osteoclastic bone resorption and to clarify the mechanism underlying this action. Methods Primary osteoclast-like cells (OLCs) were isolated from 3 week-old mice and induced by 1,25(OH) 2D 3. Then OLCs were exposed to genistein at various concentration of 0,10 -9,10 -8,10 -7,10 -6 and 10 -5mol/L. The number of TRAP+cells were counted as well as the surface area of bone resorption on bone slice.Additionally the osteoblasts were obtained from 5 day old mice and were exposed to 0, 10 -8,10 -7,10 -6 and 10 -5mol/L genistein and 10 -9mol/L 17β-estradiol. The mRNA levels of RANKL and OPG were assayed by RT-PCR. Results In vitro, the number of TRAP+cells decreased depending on the concentration of genistein as well as the area of bone resorption. By semiquantitative RT-PCR, genistein increased the mRNA levels of OPG and RANKL, and finally enlarged the ratio between them in dose and time-dependent mamer. Conclusion Genistein inhibits OLCs' bone resorption through suppressing the differentitation of preosteoclasts, and the ratio between OPG and RANKL may be the key factor.
Objective Osteoporosis and periodontal disease animal models were used to evaluate the anti-resorption effect of alendronate on bone.Methods To establish osteoporosis and alveolar bone resorption animal models.The experimental groups (ovariectomized dental ligature group and simple dental ligature group) were given alendronate in 0.5 mg/kg BW by subcutaneous injection after six weeks of ovariectomy and dental ligature,three times a week for 6 weeks.All the animals were sacrifised 12 weeks after the operation.The blood samples were collected for determination of biochemical indexes.The left femora and jaw bone were processed for histomorphometry.The right femora and mendibles were prepared for determination of bone density and bone biodynamics.Results As compared with the control groups (without alendronate),the values of experimental groups (with alendronate) were all significantly improved on bone density and bone anti-curve power.The alkaline phosphatase,calcium in serum of ovariectomy groups (without alendronate) were obviously increased.In the alendronate group,however,these indexes were close to normal levels.In histomorphometry,there were little inflammation in gingiva and less level alveolar bone resorption in the experimental groups;but there was obvious gingivitis and level alveolar bone resorption in the control groups.Conclusion Alendronate can effectively treat osteoporosis,increase bone volume and prevent bone lose in ovariectomized female rats and inhibit pathological alveolar bone resorption.
Objective To build an immortalized osteoclast cell line by the method of transgene.Methods The osteoclast precursors were derived from the mouse bone marrow cells in the presence of 1,25(OH) 2 D 3 in vitro. Transfection of two plasmids(SV40 and GFP)into the above cells respectively was performed using liposome(Fugene 6).G418 was used to select the resistant cell clones.At the same time,the GFP plasmid was introduced into the retrovirus packing cells(PT67) as a contrast.Then the osteoclast precursors were infected using the retrovirus containing GFP.G418 was used to select the resistant cell clones.Results Firstly,no G418 resistant cell cloned after the transfection of the GFP and SV40 into osteoclast proecursors by Fugene 6.But the resistant cell cloned after transfection of GFP into PT67 cells by Fugene 6.Secondly,no positive infected cell cloned in the presence of the retrovirus containing GFP.Conclusions The cell immortalization by means of transgene is a plausible way to obtain the osteoclast cell lines although it is challenging.Liposome and retrovirus vector are not the best choice for osteoclasts to transgene.\;
The character of PI Qiao-lin fails to extricate himself from constant companion of bewilderment. He wants to withstand nihilism but he cannot find the sense of life. As a the civilized man and useless man, he has a vivid character of relativity about tropism of life worthiness and moral illegibility. The personality set out bright modernity.
Objective To investigate the gene expression of osteoprotegerin (OPG) and osteoclast differentiation factor (ODF/TRANCE/RANKL), two new members of the TNF-receptor superfamily, in giant cell tumor (GCT); to discuss the molecular mechanism of extensive bone resorption caused by GCT. Methods Using TRIzol reagent to prepare total RNA from GCT sample and normal bone tissue. By a first-strand complementary DNA (cDNA) synthesis kit, cDNA was synthesized from 2.0 μg RNA according to the manufacturer′s instructions. cDNA was then amplified by PCR . Amplification products were resolved by electrophoresis on a 1.5% agarose gel and stained with EB. The relative quantity of the PCR products were determined and the mRNA levels of OPG, ODF, M-CSF (cofactor of ODF), and RANK (receptor of ODF) were compared with that of the normal bone. Results GCT contained highly expressed mRNA of ODF , OPG, M-CSF and RANK. There was mRNA expression of OPG , M-CSF and RANK and less expression of ODF in normal bone. The ODF mRNA and RANK mRNA in GCT were more abundant than that in normal bone. In GCT, the ratio of ODF mRNA exceeded OPG expression. But in normal bone, the OPG mRNA exceeded ODF expression. Conclusions The results suggest that GCT contains all signals including OPG, ODF, M-CSF and RANK that are essential for inducing osteoclastogenesis and promoting bone resorption.
Study on overvoltage and insulation coordination can provide basic technical support for the implementation of Zhoushan multi-terminals VSC-HVDC( voltage sourced converters based high voltage direct current) project. In order to determine the protection configuration of arrester scheme and the insulation level of equipment,integrated design was carried out and the main circuit parameters of the project were proposed,as well as the main wiring. According to the control strategy of Zhoushan multi-terminals VSC-HVDC project,the electromagnetic transient simulation model was established. The capacitance of sub-module in converters,the type of transformer,the inductance of arm reactors and DC reactors,the grounding modes and the selection of cable parameters all influenced simulation results significantly in the model. Based on the detailed research,the equipments' parameters were selected and used for the overvoltage simulation,the insulation levels of main equipments were determined as well. As the key equipment in this project,the DC cable's insulation margin was suggested to be 30% in order to ensure the safe operation,and its insulation level was finally determined to be 510 kV. The research results have been applied in Zhoushan multi-terminals VSC-HVDC project,which can provide references for other similar projects.
Objective To study the character of lymphocytes in benign lymphoadenosis of oral mucosa(BLOM).Methods The expression of SP immunohistochemical staining against CD20, CD45RO were evaluated in 14 cases of BLOM,9 BLOM with dysplasia, 11 carcinogenesis of BLOM and 10 normal oral mucosa.Results There was CD45RO positive staining in 8 out of 10 normal tissues and there was no CD20 positive staining in 10 normal tissues. There was CD20 negative staining in 1 out of 11 carcinogenesis of BLOM and there were simultaneously CD20 and CD45RO positive staining in all the other BLOM.Conclusion the lymphocytes of BLOM came from both T cell and B cell. This proved the lymphoid tissue in BLOM is mucosa-associated lymphoid tissue (MALT), it is not a kind of lymphoma but a kind of proliferation disease.
Objective To investigate the possible nature of the multinucleated giant cells (MGCs) in giant cell tumor (GCT) and giant cell granuloma (GCG) of the jaws.Methods Eight cases of GCT and 8 cases of GCG [4 central cases (CGCG) and 4 peripheral cases (PGCG)],1 case of scrofula and 1 case of foreign body giant cell granuloma were undergone immunohistochemical staining to detect CD68, AAT,AACT,lysozyme,MAC-387,vimentin,S-100 and Ki-67 using formalin-paraffin embedded tissue sections.Enzyme histochemical staining for tartrate-resistant acid phosphatase (TRAP),an enzyme marker for osteoclasts,was also performed in all cases.Results MGCs in GCT and GCG lesions possessed both histiocyte/monocyte related antigens (CD68,AAT,AACT) as well as osteoclast specific enzyme (TRAP).Ki67 staining,however,showed that only mononuclear cells,but not the MGCs,were in cell cycle.Conclusion The MGCs in GCT and GCG appeared to express both macrophage and osteoclast associated phenotypes.The exclusively proliferative activity demonstrated in the mononuclear cells and the similar immunocytochemical and histochemical expression in some of the mononuclear elements suggest that these cells may represent precursors of the MGCs.Coalescence or fusion of these precursor cells could result in giant cell formation in the lesion.
Objective To detect SH3BP2 gene mutation in a cherubism family.Methods Peripheral blood samples were obtained from the family of cherubism. Geomic DNA was extracted. Polymerase chain reaction and direct sequencing were performed to identify the mutation.Results A transition in exon 9 in SH3BP2 gene was detected in the family, which led to a missense mutation(Arg 415 Pro). Conclusions Missense mutation in the SH3BP2 gene was responsible for the phenotypes of this Chinese cherubism family.
