s-Lap is a new gene sequence from pig retinal pigment epithelial(RPE) cells, which was found and cloned in the early period of apoptosis of RPE cells damaged with visible light. We cloned the coding area sequence of t...s-Lap is a new gene sequence from pig retinal pigment epithelial(RPE) cells, which was found and cloned in the early period of apoptosis of RPE cells damaged with visible light. We cloned the coding area sequence of the novel gene of s-Lap and constructed its recombinant eukaryotic plasmid pcDNA3.1-GFP/s-lap with the recombinant DNA technique. The expression and localization of s-lap/GFP fusion protein in CHO and B_~16 cell lines were studied with the instantaneously transfected pcDNA3.1-GFP/s-lap recombinant plasmid. ~s-Lap/GFP fusion protein can be expressed in CHO and B_~16 cells with a high rate expression in the nuclei.展开更多
In model experiments were studied the effect of cosmo-geophysical factors of environment (hypomagnetic conditions during 2 days ≈ 1 mkT;electromagnetic irradiation (10 min - 2 MHz with amplitude 5 V/m and power 30 mk...In model experiments were studied the effect of cosmo-geophysical factors of environment (hypomagnetic conditions during 2 days ≈ 1 mkT;electromagnetic irradiation (10 min - 2 MHz with amplitude 5 V/m and power 30 mkVt, background 2 - 4 mkVt), γ-quantum (10 min—from the source 137Cs) and its combined effect on the physic-chemical properties (ORP and pH) of growing medium for cell culture of mammals as nutrition medium 199 (PanEco, Russia). It was used a clear solution of medium (solution 1) and with the adding of 10% embryo bull serum—model of bio-medium (solution 2). Hypomagnetic conditions evoked the decreasing of ORP and pH value in both solutions, electromagnetic irradiation in the solution 1 which evoked the decreasing of ORP and the increasing of pH value, and in the solution 2, on the contrary, the increasing of ORP with the unchanging pH value. γ-radiation sharply decreased ORP value and didn’t change pH in solution 1, i.e. the reduction properties increased. There is insignificant increasing of ORP value and the decreasing of pH is noted in the solution 2, that it is characterized with the increasing of oxidative properties of solution. Under the combined effect of hypomagnetic conditions and electromagnetic irradiation, the values of investigating parameters in the solution 1 decreased and in the solution 2 increased. It was observed acute decreasing of ORP value in both solutions under the combined effect of hypomagnetic conditions and γ-radiation, i.e. the reductive properties of the solutions increased sharply. In this the concentration H+ significantly decreased, (p γ-radiation led to the decreasing of ORP and pH values in both solutions. Thus, the studying factors significantly change the oxidation-reduction properties of growing mediums. The investigation of the processes in biological mediums plays the important role in the assessment of environment effect during the flight in inter-planet space.展开更多
Lead ion （Pb2＋） has been proven to be a neurotoxin due to its neurotoxicity on mammalian nervous system, especially for the developing brains of juveniles. However, many reported studies involved the negative effec...Lead ion （Pb2＋） has been proven to be a neurotoxin due to its neurotoxicity on mammalian nervous system, especially for the developing brains of juveniles. However, many reported studies involved the negative effects of Pb2＋ on adult neural cells of humans or other mammals, only few of which have examined the effects of Pb2＋ on neural stem cells. The purpose of this study was to reveal the biological effects of Pb2＋from lead acetate [Pb （0H30OO）2] on viability, proliferation and differentiation of neural stem cells derived from the hippocampus of newborn rats aged 7 days and adult rats aged 90 days, respectively. This study was carried out in three parts. In the first part, 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide assay （MTT viability assay） was used to detect the effects of Pb2＋ on the cell viability of passage 2 hippocampal neural stem cells after 48-hour exposure to 0-200 pM Pb2＋. In the second part, 10 pM bromodeoxyuridine was added into the culture medium of passage 2 hippocampal neural stem cells after 48-hour exposure to 0- 200 pM Pb2＋, followed by immunocytochemical staining with anti-bromodeoxyuridine to demonstrate the effects of Pb2＋ on cell proliferation. In the last part, passage 2 hippocampal neural stem cells were allowed to grow in the differentiation medium with 0-200 pM Pb2＋. Immunocytochemical staining with anti-microtubule-associated protein 2 （a neuron marker）, anti-glial fibrillary acidic protein （an astrocyte marker）, and anti-RIP （an oligodendrocyte marker） was performed to detect the differentiation commitment of affected neural stem cells after 6 days. The data showed that Pb2~ inhibited not only the viability and proliferation of rat hippocampal neural stem cells, but also their neuronal and oligodendrocyte differentiation in vitro. Moreover, increased activity of astrocyte differentiation of hippocampal neural stem cells from both newborn and adult rats was observed after exposure to high concentration of lead ion in vitro. These findings suggest that hippocampal neural stem cells of newborn rats were more sensitive than those from adult rats to Pb2＋cytotoxicity.展开更多
文摘s-Lap is a new gene sequence from pig retinal pigment epithelial(RPE) cells, which was found and cloned in the early period of apoptosis of RPE cells damaged with visible light. We cloned the coding area sequence of the novel gene of s-Lap and constructed its recombinant eukaryotic plasmid pcDNA3.1-GFP/s-lap with the recombinant DNA technique. The expression and localization of s-lap/GFP fusion protein in CHO and B_~16 cell lines were studied with the instantaneously transfected pcDNA3.1-GFP/s-lap recombinant plasmid. ~s-Lap/GFP fusion protein can be expressed in CHO and B_~16 cells with a high rate expression in the nuclei.
文摘In model experiments were studied the effect of cosmo-geophysical factors of environment (hypomagnetic conditions during 2 days ≈ 1 mkT;electromagnetic irradiation (10 min - 2 MHz with amplitude 5 V/m and power 30 mkVt, background 2 - 4 mkVt), γ-quantum (10 min—from the source 137Cs) and its combined effect on the physic-chemical properties (ORP and pH) of growing medium for cell culture of mammals as nutrition medium 199 (PanEco, Russia). It was used a clear solution of medium (solution 1) and with the adding of 10% embryo bull serum—model of bio-medium (solution 2). Hypomagnetic conditions evoked the decreasing of ORP and pH value in both solutions, electromagnetic irradiation in the solution 1 which evoked the decreasing of ORP and the increasing of pH value, and in the solution 2, on the contrary, the increasing of ORP with the unchanging pH value. γ-radiation sharply decreased ORP value and didn’t change pH in solution 1, i.e. the reduction properties increased. There is insignificant increasing of ORP value and the decreasing of pH is noted in the solution 2, that it is characterized with the increasing of oxidative properties of solution. Under the combined effect of hypomagnetic conditions and electromagnetic irradiation, the values of investigating parameters in the solution 1 decreased and in the solution 2 increased. It was observed acute decreasing of ORP value in both solutions under the combined effect of hypomagnetic conditions and γ-radiation, i.e. the reductive properties of the solutions increased sharply. In this the concentration H+ significantly decreased, (p γ-radiation led to the decreasing of ORP and pH values in both solutions. Thus, the studying factors significantly change the oxidation-reduction properties of growing mediums. The investigation of the processes in biological mediums plays the important role in the assessment of environment effect during the flight in inter-planet space.
基金supported by a grant from the University of Hong Kong, China
文摘Lead ion （Pb2＋） has been proven to be a neurotoxin due to its neurotoxicity on mammalian nervous system, especially for the developing brains of juveniles. However, many reported studies involved the negative effects of Pb2＋ on adult neural cells of humans or other mammals, only few of which have examined the effects of Pb2＋ on neural stem cells. The purpose of this study was to reveal the biological effects of Pb2＋from lead acetate [Pb （0H30OO）2] on viability, proliferation and differentiation of neural stem cells derived from the hippocampus of newborn rats aged 7 days and adult rats aged 90 days, respectively. This study was carried out in three parts. In the first part, 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide assay （MTT viability assay） was used to detect the effects of Pb2＋ on the cell viability of passage 2 hippocampal neural stem cells after 48-hour exposure to 0-200 pM Pb2＋. In the second part, 10 pM bromodeoxyuridine was added into the culture medium of passage 2 hippocampal neural stem cells after 48-hour exposure to 0- 200 pM Pb2＋, followed by immunocytochemical staining with anti-bromodeoxyuridine to demonstrate the effects of Pb2＋ on cell proliferation. In the last part, passage 2 hippocampal neural stem cells were allowed to grow in the differentiation medium with 0-200 pM Pb2＋. Immunocytochemical staining with anti-microtubule-associated protein 2 （a neuron marker）, anti-glial fibrillary acidic protein （an astrocyte marker）, and anti-RIP （an oligodendrocyte marker） was performed to detect the differentiation commitment of affected neural stem cells after 6 days. The data showed that Pb2~ inhibited not only the viability and proliferation of rat hippocampal neural stem cells, but also their neuronal and oligodendrocyte differentiation in vitro. Moreover, increased activity of astrocyte differentiation of hippocampal neural stem cells from both newborn and adult rats was observed after exposure to high concentration of lead ion in vitro. These findings suggest that hippocampal neural stem cells of newborn rats were more sensitive than those from adult rats to Pb2＋cytotoxicity.
