OBJECTIVE: To observe the distribution of copper in the subcellular structure for the understanding of primary pathogenesis of hepatolenticular degeneration (HLD). METHODS: Skin fibroblasts taken from HLD patients wer...OBJECTIVE: To observe the distribution of copper in the subcellular structure for the understanding of primary pathogenesis of hepatolenticular degeneration (HLD). METHODS: Skin fibroblasts taken from HLD patients were cultured as an in vitro model of HLD, and the control cells taken from healthy volunteers were clutured in the same way. The distribution of copper inside and outside of lysosomes in fibroblasts was detected by quantitative electron probe X-ray microanalysis. The relationship between the subcellular location of copper and the genotype of the patients, and relationship between the distribution of copper and the course of the disease were analyzed. RESULTS: The content of Cu^(2+) inside lysosomes of HLD cells (14.6±2.1 mmol/kg) and of heterozygote cells (11.6±0.6 mmol/kg) was higher than that of normal cells (4.5±1.2 mmol/kg) (P<0.01). The content of Cu^(2+) outside lysosomes of HLD cells (17.5±4.2 mmol/kg) and of heterozygote cells (12.0±0.9 mmol/kg) was higher than that of normal cells (4.7±1.2 mmol/kg) (P<0.01). The distribution of copper in the subcellular structure was correlated with disease courses of HLD patients. With the progression of the disease, more copper was deposited in lysosomes (r=0.85, P<0.01). The content of copper in the diffused cytoplasmic compartment in HLD cells was correlated with that of sulfur (r=0.86, P<0.05), but not in heterozygote and normal cells. CONCLUSIONS: In the early stage of HLD, copper is accumulated outside lysosome, which is paralleled with increase of metallothionein-like proteins (copper and sulfur-binding proteins). With the development of the disease, more copper is deposited inside lysosome than outside lysosome. We conclude that the up-regulation expression of copper and sulfur-binding proteins and copper accumulation in lysosomes may play an important role in lowering the ATP7B gene mutation-induced toxic effects of free copper on the cell.展开更多
The effect of operating conditions on the time-dependent X-ray intensity variation is of great importance for the optimal EPMA conditions for accurate determinations of various elements in carbonate minerals. Beam dia...The effect of operating conditions on the time-dependent X-ray intensity variation is of great importance for the optimal EPMA conditions for accurate determinations of various elements in carbonate minerals. Beam diameters of 0, 1, 2, 5, 10, 15, and 20 μm, and beam currents of 3, 5, 10, 20,and 50 nA were tested. Ca, Mg, Zn, and Sr were found to be more sensitive to electron beam irradiation as compared to other elements, and small currents and large beam diameters minimized the timedependent X-ray intensity variations. We determined the optimal EPMA operating conditions for elements in carbonate: 10 μm and 5 nA for calcite;10 μm and 10 nA for dolomite;5 μm and 10 nA or 10 μm and 20 nA for strontianite;and 20 nA and 5 μm for other carbonate. Elements sensitive to electron beam irradiation should be determined first. In addition, silicate minerals are preferred as standards rather than carbonate minerals.展开更多
AIM To develop methods of articular cartilage preparation for X-ray-electron probe microanalysis and to study its elements content in experimental osteoarthrosis.METHODS Twenty dogs aged 2-8 years were divided in rese...AIM To develop methods of articular cartilage preparation for X-ray-electron probe microanalysis and to study its elements content in experimental osteoarthrosis.METHODS Twenty dogs aged 2-8 years were divided in research(aged 2 years, induction of osteoarthrosis-IOA) and intact group. Intact group included three subgroups(aged 2, 5 and 8 years). Samples of cartilage after araldite saturation and pouring were partially cut into semithin sections stained with methylene blue and with methylene blue-basic fuchsin. Their smooth surfaces were investigated by X-ray-electron probe microanalysis. Spatial distribution of sulfur, calcium and phosphorus and their concentrations(weight %) were investigated.RESULTS X-ray electron probe microanalysis revealed non-uniformsulfur distribution in cartilage of intact animals: Its content increases from superficial zone to deep one, this regularity was preserved in animals with IOA. Differences of IOA with spontaneous chondropathy were revealed. Spontaneous aging was characterized by calcium and phosphorus storage in deep and calcified zones and compensatory increase of sulfated glycosaminoglycans in intermediate and deep cartilage zones as evidenced by the metachromatic reaction and microanalysis data. Unlike spontaneous chondropathy connected with aging in experimentally stimulated osteoarthrosis more intensive storage of calcium but minor phosphorus in intermediate zone were marked. In IOA the calcified cartilage thinning and osteoclastic resorption are apparent with few changes of elements composition; the only difference from control is minority phosphorus content.CONCLUSION The obtained results demonstrate specific tricks of X-ray electron probe microanalysis and its possibility in the research of mechanisms of articular cartilage alterations in osteoarthrosis.展开更多
In this paper, a simple and facile surfactant assisted combustion synthesis is reported for the ZnO nanoparticles. The synthesis of ZnO-NPs has been done with the assistance of non-ionic surfactant TWEEN 80. The effec...In this paper, a simple and facile surfactant assisted combustion synthesis is reported for the ZnO nanoparticles. The synthesis of ZnO-NPs has been done with the assistance of non-ionic surfactant TWEEN 80. The effect of fuel variations and comparative study of fuel urea and glycine have been studied by using characterization techniques like X-ray diffraction (XRD), transmission electron microscope (TEM) and particle size analyzer. From XRD, it indicates the presence of hexagonal wurtzite structure for ZnO-NPs. Using X-ray broadening, crystallite sizes and lattice strain on the peak broadening of ZnO-NPs were studied by using Williamson-Hall (W-H) analysis and size-strain plot. Strain, stress and energy density parameters were calculated for the XRD peaks of all the samples using (UDM), uniform stress deformation model (USDM), uniform deformation energy density model (UDEDM) and by the size-strain plot method (SSP). The results of mean particle size showed an inter correlation with W-H analysis, SSP, particle analyzer and TEM results.展开更多
文摘OBJECTIVE: To observe the distribution of copper in the subcellular structure for the understanding of primary pathogenesis of hepatolenticular degeneration (HLD). METHODS: Skin fibroblasts taken from HLD patients were cultured as an in vitro model of HLD, and the control cells taken from healthy volunteers were clutured in the same way. The distribution of copper inside and outside of lysosomes in fibroblasts was detected by quantitative electron probe X-ray microanalysis. The relationship between the subcellular location of copper and the genotype of the patients, and relationship between the distribution of copper and the course of the disease were analyzed. RESULTS: The content of Cu^(2+) inside lysosomes of HLD cells (14.6±2.1 mmol/kg) and of heterozygote cells (11.6±0.6 mmol/kg) was higher than that of normal cells (4.5±1.2 mmol/kg) (P<0.01). The content of Cu^(2+) outside lysosomes of HLD cells (17.5±4.2 mmol/kg) and of heterozygote cells (12.0±0.9 mmol/kg) was higher than that of normal cells (4.7±1.2 mmol/kg) (P<0.01). The distribution of copper in the subcellular structure was correlated with disease courses of HLD patients. With the progression of the disease, more copper was deposited in lysosomes (r=0.85, P<0.01). The content of copper in the diffused cytoplasmic compartment in HLD cells was correlated with that of sulfur (r=0.86, P<0.05), but not in heterozygote and normal cells. CONCLUSIONS: In the early stage of HLD, copper is accumulated outside lysosome, which is paralleled with increase of metallothionein-like proteins (copper and sulfur-binding proteins). With the development of the disease, more copper is deposited inside lysosome than outside lysosome. We conclude that the up-regulation expression of copper and sulfur-binding proteins and copper accumulation in lysosomes may play an important role in lowering the ATP7B gene mutation-induced toxic effects of free copper on the cell.
基金supported by the Natural Science Foundation of China (No. 41403022)the Fundamental Research Funds for the Central Universities, China University of Geosciences (Wuhan) (No. CUGL150401)
文摘The effect of operating conditions on the time-dependent X-ray intensity variation is of great importance for the optimal EPMA conditions for accurate determinations of various elements in carbonate minerals. Beam diameters of 0, 1, 2, 5, 10, 15, and 20 μm, and beam currents of 3, 5, 10, 20,and 50 nA were tested. Ca, Mg, Zn, and Sr were found to be more sensitive to electron beam irradiation as compared to other elements, and small currents and large beam diameters minimized the timedependent X-ray intensity variations. We determined the optimal EPMA operating conditions for elements in carbonate: 10 μm and 5 nA for calcite;10 μm and 10 nA for dolomite;5 μm and 10 nA or 10 μm and 20 nA for strontianite;and 20 nA and 5 μm for other carbonate. Elements sensitive to electron beam irradiation should be determined first. In addition, silicate minerals are preferred as standards rather than carbonate minerals.
基金Supported by The RF Ministry of Health within government-mandated program for FSBI Russian Ilizarov Scientific Center "Restorative Traumatology and Orthopaedics"(RISC"RTO") for Scientific Research,No.01201155770
文摘AIM To develop methods of articular cartilage preparation for X-ray-electron probe microanalysis and to study its elements content in experimental osteoarthrosis.METHODS Twenty dogs aged 2-8 years were divided in research(aged 2 years, induction of osteoarthrosis-IOA) and intact group. Intact group included three subgroups(aged 2, 5 and 8 years). Samples of cartilage after araldite saturation and pouring were partially cut into semithin sections stained with methylene blue and with methylene blue-basic fuchsin. Their smooth surfaces were investigated by X-ray-electron probe microanalysis. Spatial distribution of sulfur, calcium and phosphorus and their concentrations(weight %) were investigated.RESULTS X-ray electron probe microanalysis revealed non-uniformsulfur distribution in cartilage of intact animals: Its content increases from superficial zone to deep one, this regularity was preserved in animals with IOA. Differences of IOA with spontaneous chondropathy were revealed. Spontaneous aging was characterized by calcium and phosphorus storage in deep and calcified zones and compensatory increase of sulfated glycosaminoglycans in intermediate and deep cartilage zones as evidenced by the metachromatic reaction and microanalysis data. Unlike spontaneous chondropathy connected with aging in experimentally stimulated osteoarthrosis more intensive storage of calcium but minor phosphorus in intermediate zone were marked. In IOA the calcified cartilage thinning and osteoclastic resorption are apparent with few changes of elements composition; the only difference from control is minority phosphorus content.CONCLUSION The obtained results demonstrate specific tricks of X-ray electron probe microanalysis and its possibility in the research of mechanisms of articular cartilage alterations in osteoarthrosis.
文摘In this paper, a simple and facile surfactant assisted combustion synthesis is reported for the ZnO nanoparticles. The synthesis of ZnO-NPs has been done with the assistance of non-ionic surfactant TWEEN 80. The effect of fuel variations and comparative study of fuel urea and glycine have been studied by using characterization techniques like X-ray diffraction (XRD), transmission electron microscope (TEM) and particle size analyzer. From XRD, it indicates the presence of hexagonal wurtzite structure for ZnO-NPs. Using X-ray broadening, crystallite sizes and lattice strain on the peak broadening of ZnO-NPs were studied by using Williamson-Hall (W-H) analysis and size-strain plot. Strain, stress and energy density parameters were calculated for the XRD peaks of all the samples using (UDM), uniform stress deformation model (USDM), uniform deformation energy density model (UDEDM) and by the size-strain plot method (SSP). The results of mean particle size showed an inter correlation with W-H analysis, SSP, particle analyzer and TEM results.
