Two new ent-kauranoids, tenuifolin A (3beta,6alpha, 15beta-trihydroxy-1alpha, 7beta-diacetoxy-11beta, 16beta-epoxy-ent-kaurane) (1) and tenuifolin B (1alpha,6alpha, 11beta-trihydroxy-3beta,7beta-diacetoxy-ent-kaur-16-...Two new ent-kauranoids, tenuifolin A (3beta,6alpha, 15beta-trihydroxy-1alpha, 7beta-diacetoxy-11beta, 16beta-epoxy-ent-kaurane) (1) and tenuifolin B (1alpha,6alpha, 11beta-trihydroxy-3beta,7beta-diacetoxy-ent-kaur-16-en-15-one) (2), together with four known compounds were isolated from the aerial parts of Isodon tenuifolia (W. W. Smith) Kudo collected from Zhongdian County, Yunnan Province, China. Their structures were determined by the spectral methods (including 2D NMR techniques).展开更多
Northern blot analysis was conducted with mitochondrial RNA from seedling leaves, floral buds, and developing seeds of NCa CMS, maintainer line and fertile F1 using ten mitochondrial genes as probes. The results revea...Northern blot analysis was conducted with mitochondrial RNA from seedling leaves, floral buds, and developing seeds of NCa CMS, maintainer line and fertile F1 using ten mitochondrial genes as probes. The results revealed that 9 out of the 10 mitochondrial genes, except for atp6, showed no difference in different tissues of the corresponding materials of NCα CMS system and that they might be constitutively expressed genes. Eight genes, such as orf139, orf222, atpl, cox1, cox2, cob, rm5S, and rm26S, showed no difference among the three tissues of all the materials detected. So the expression of these eight genes was not regulated by nuclear genes and was not tissue-specific. The transcripts of atp9 were identical among different tissues, but diverse among different materials, indicating that transcription of atp9 was neither controlled by nuclear gene nor tissue-specific. Gene atp6 displayed similar transcripts with the same size among different tissues of all the materials but differed in abundance among tissues of corresponding materials and its expression might be tissue-specific under regulation of nuclear gene. Moreover, three transcripts of orf222 were detected in the floral buds of NCa cms and fertile F1, but no transcript was detected in floral buds of the maintainer line.The transcription of orf139 was similar to that of orf222 but only two transcripts of 0.8 kb and 0.6 kb were produced. The atp9 probe detected a single transcript of 0.6 kb in NCa cms and in maintainer line and an additional transcript of 1.2 kb in fertile F1. The relationship of expression of orf222, orf139, and atp9 with NCa sterility was discussed.展开更多
The ultrastructure of the vegetative cells of Nostoc flagelliforme Born. et Flah. was investigated with high pressure freezing and freeze substitution technique and compared with the results obtained by using conv...The ultrastructure of the vegetative cells of Nostoc flagelliforme Born. et Flah. was investigated with high pressure freezing and freeze substitution technique and compared with the results obtained by using conventional preparation methods. During the processes of chemical fixation, dehydration and embedding, the cell structures might be more artificially modified than that obtained from high pressure freezing and freeze substitution. With the present method, the sheath of N. flagelliforme could be well penetrated and no extra big space could exist between the cell and the sheath. The cell protoplasm rarely shrinked. Some fine structures of cell inclusions and unit membranes became visualized. Many bacteria were harbored in the sheath. In addition, the presence of big vacuoles in the cell of N. flagelliforme as well as the presence of bacteria in the sheath shown in the present preparation for cyanobacteria has not been described so far in the literature.展开更多
[Objective] This study aimed to investigate the feasibility of agricultural irri- gation with reclaimed water in Urumqi City. [Method] Chinese cabbages were used as experimental materials and irrigated with control wa...[Objective] This study aimed to investigate the feasibility of agricultural irri- gation with reclaimed water in Urumqi City. [Method] Chinese cabbages were used as experimental materials and irrigated with control water, 50% reclaimed water and 100% reclaimed water, and then the number of endophytic bacteria in Chinese cab- bages was measured. [Result] Using 50% reclaimed water, the fresh weight of Chi- nese cabbage was improved by 68.94%; however, with the deepening internalization of exogenous microorganism internalization, the total number of endophytic bacteria and coliform flock in Chinese cabbages treated with 50% reclaimed water was sig- nificantly higher than the other two treatment groups (P〈0.05). [Conclusion] Re- claimed water enhances both the weight of Chinese cabbage and the number of endophytic bacteria, which is unsuitable for irrigation of edible crops such as Chi- nese cabbage. Strengthening disinfection during the process of sewerage treatment could solve this problem.展开更多
Cytochrome b_559 in photosystem Ⅱ reaction center was purified from spinach ( Spinacia oleracea L.) and rice ( Oryza sativa L.) by a rapid and simple procedure. Their low temperature fluorescence emission and e...Cytochrome b_559 in photosystem Ⅱ reaction center was purified from spinach ( Spinacia oleracea L.) and rice ( Oryza sativa L.) by a rapid and simple procedure. Their low temperature fluorescence emission and excitation spectra, ultraviolet fluorescence spectra and absolute absorption spectra were presented. The author's purification methods, which enhanced the yield of pure protein and shorted the time for isolation, have several advantages: 1. use of oxygen_evolving PSⅡ core complexes as the starting material in order to avoid disturbing from other cytochromes; 2. isocratic elution of cytochrome b_559 from a DEAE_Sephacel column for eliminating the impurity and yielding the protein in pure state; 3. a simple column procedure for removal of excess Triton X_100. Purified cytochromes b_559 from these species have similar optical spectra and mobility during gel electrophoresis under native conditions. From the results of novel electrophoresis (Tricine_SDS_PAGE), cytochrome b_559 from both spinach and rice reveal two polypeptide bands (apparent molecular weight 9 kD and 4 kD, respectively). By measuring of 77 K fluorescence spectra, it was shown that for the purified cytochrome b_559 there were two excitation peaks at 439 nm and 413 nm, and two emission peaks at 563 nm and 668 nm. This is the first indication that Cyt b_559 is able to emit fluorescence and also transfer excited electrons to chlorophyll. By the use of ultraviolet fluorescence spectra, it was demonstrated for the first time that the location of Trp residue could be in the hydrophobic transmembrane region of cytochrome b_559.展开更多
Abstract [Objective] The paper was to prepare and detect the extended DNA fibers of Chinese cabbage. [Method] Chinese cabbage nuclei was first successfully isolated by chopping young leaves with a blade, then nuclei w...Abstract [Objective] The paper was to prepare and detect the extended DNA fibers of Chinese cabbage. [Method] Chinese cabbage nuclei was first successfully isolated by chopping young leaves with a blade, then nuclei were lysed by SDS to release DNA, and DNA fibers were dragged and extended with a coverslip. [Result] The results of Fiber-FISH with genomic DNA and 25S rDNA as probes showed that DNA fiber size as long as about 1.93 Mb could be measured and the number of 25S rDNA copies region were estimated to be 258 and 687 in Chinese cabbage genome. DNA fibers prepared by this method showed equally spread parallel thread with clear background, and were suitable for FISH analysis. [Conclusion] The study would accelerate Chinese cabbage genome mapping and organization analysis.展开更多
[Objective] The aim was to study the reproduction of the three-line genic male sterile (GMS) lineparent Mian7MB-1 (B. NapusL.) and the seed production of F1 through somatic tissue culture. [Methed] Through hybridi...[Objective] The aim was to study the reproduction of the three-line genic male sterile (GMS) lineparent Mian7MB-1 (B. NapusL.) and the seed production of F1 through somatic tissue culture. [Methed] Through hybridization, a new breeding material Mian 7MB-1 in three-line genic temporary maintainer line propagated by tissue culture was used to improve the sterile plant rate of rapeseed in dual-purpose recessive GMS line, such as Mian 7AB type, S45AB type, and etc. And then the variety comparative test was performed. [Result] In order to avoid some fertility restoration phenomena occurring during the process of self-reproduction, Mian 7AB was propagated in bulk with somatic tissue culture of temporary maintainer line plant stem. The propagated temporary maintainer line seedlings were applied to the breeding and seed production of net room male sterile line parent, promoting the sterile plant rate of the male sterile line parent to 91.7% -93.5%. The male sterile line parents per hectare were enough for the seed production of hybrid F1 in 7 500 -15 000 hm^2. [ Conclusion ] Compared with the original dual-purpose GMS line, the seed production ultilizing male sterile line with high sterile plant rate greatly reduced the labor, significantly improved the seed yield, ensuring the seed quality and forming a perfect breeding and seed production system.展开更多
It had been demonstrated that individual cells or protoplasts isolated fromPorphyrathallus by enzyme could develop into normal leafy thalli in the same way as monospores, and that isolated cells develop in different w...It had been demonstrated that individual cells or protoplasts isolated fromPorphyrathallus by enzyme could develop into normal leafy thalli in the same way as monospores, and that isolated cells develop in different way in liquid and on semi-solid media. The authors observed the ultrastructure of isolated vegetative cells cultured on semi-solid media and compared them with those of monospores and isolated cells cultured in liquid media. The results showed that subcellular structures were quite different among cells in different conditions. In their development, isolated cells on semi-solid media did not show the characteristic subcellular feature of monospore formation, such as production of fibrous vesicles. Callus-like cells formed on semi-solid media underwent a distinctive modification in cellular organization. They developed characteristic cell inclusions and a special 2-layer cell covering. Golgi bodies, ER, starch grains, mitochondria. Vacuoles were not commonly found in them.展开更多
[ Objective] The study is to generate pharmaceutical protein via plant transgenic technique. [Methed] Using the cotyledons with petiole as transformation receptor, the fusion gene of rapeseed oil-body gene and bFGF wa...[ Objective] The study is to generate pharmaceutical protein via plant transgenic technique. [Methed] Using the cotyledons with petiole as transformation receptor, the fusion gene of rapeseed oil-body gene and bFGF was introduced into the rapeseed ( Brassica campestris L. ) by Agrobacterium tumefaciens-mediated transformation; meanwhile regeneration conditions of rapeseed were also optimized, and the regenerated resistant plantlets were detected by PCR and Southern blot. [ Result] This fusion gene had been integrated into rapeseed genome successfully, and the optimized conditions of transformation and regeneration were as follows: explants pre-culture for 2 d, co-culture for 3 d, bacteria solution OD600 for 0.3 and infection time for 5 min. [ Conclusion] The results laid a solid foundation for extraction, isolation and purification of protein in transgenic plant seeds.展开更多
AIM: To investigate the potential effects of chelidonine, the main alkaloid of Chelidonium majus, on telomerase activity and its regulation in HepG2 cells. METHODS: Cytotoxicity of chelidonine for HepG2 cells was de...AIM: To investigate the potential effects of chelidonine, the main alkaloid of Chelidonium majus, on telomerase activity and its regulation in HepG2 cells. METHODS: Cytotoxicity of chelidonine for HepG2 cells was determined by neutral red assay. A modified polymerase chain reaction (PCR)-based telomerase repeat amplification protocol was used to estimate relative telomerase activity in chelidonine-treated cells in comparison with the untreated control cells. Relative expression level of the catalytic subunit of telomerase (hTERT) gene and P-glycoprotein (pgp) were estimated using semi-quantitative real-time reverse transcription-PCR (RT-PCR). Cell senescence in treated cells was demonstrated using a 13-galactosidase test. RESULTS: Cytotoxicity of chelidonine in HepG2 cells was not dose-dependent and tended to reach plateau immediately after the living cells were reduced in number to slightly higher than 50%. However, 12 μmol/L concentration of chelidonine was considered as LD50, where the maximal attainable effects were realized. Real-time RT-PCR data showed that the expression of pgp increased three-fold in chelidonine treated HepG2 cells in comparison with the untreated controls. MoP phologically, treated HepG2 cells showed apoptotic features after 24 h and a small fraction of cells appeared with single blister cell death. The relative expression level of Bcl-2 dropped to less than 50% of control cells at a sub-apoptotic concentration of chelidonine and subsequently increased to higher than 120% at LD50. Telomerase activity was reduced considerably after administration of very low doses of chelidonine, whereas higher concentrations of chelidonine did not remarkably enhance the effect. Real-time RT-PCR experiments indicated a drastic decrease in expression level of hTERT subunit of telomerase under treatment with chelidonine. Repeated treatment of cells with very low doses of chelidonine caused a decline in growth rate by 4 wk and many of the cells appeared to be aged with large volume and dark staining in the β-galactosidase assay. CONCLUSION: Chelidonine reduces telomerase activity through down-regulation of hTERT expression. Senescence induction might not be directly caused by reducing telomerase activity as it occurs after a few population doublings.展开更多
AIM: To evaluate the antioxidant and phase II detoxification enzyme inducing ability of green leaf vegetables consumed in Asia.METHODS: The antioxidant properties of six commonly consumed Asian vegetables were deter...AIM: To evaluate the antioxidant and phase II detoxification enzyme inducing ability of green leaf vegetables consumed in Asia.METHODS: The antioxidant properties of six commonly consumed Asian vegetables were determined using the ABTS, DPPH, deoxyribose, PR bleaching and ironascorbate induced lipid peroxidation assay. Induce of phase II detoxification enzymes was also determined for each respective vegetable extract. Protection against authentic ONOO- and HOCl mediated cytotoxicity in human colon HCT116 cells was determined using the MTT 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrasodium bromide) viability assay.RESULTS: All of the extracts derived from green leaf vegetables exhibited antioxidant properties, while also having cytoprotective effects against ONOO- and HOCI mediated cytotoxicity. In addition, evaluation of the phase II enzyme inducing ability of each extract, as assessed by quinone reductase and glutathione- S-transferase activities, showed significant variation between the vegetables analyzed.CONCLUSION: Green leaf vegetables are potential sources of antioxidants and phase II detoxification enzyme inducers in the Asian diet. It is likely that consumption of such vegetables is a major source of beneficial phytochemical constituents that may protect against colonic damage.展开更多
The effects of different concentrations of photosynthetic bacteria solutions (CK, 0 ml photosynthetic bacteria agent+4 000 ml nutrient solution; T1, 200 ml pho- tosynthetic bacteria agent+3 800 ml nutrient solution...The effects of different concentrations of photosynthetic bacteria solutions (CK, 0 ml photosynthetic bacteria agent+4 000 ml nutrient solution; T1, 200 ml pho- tosynthetic bacteria agent+3 800 ml nutrient solution; T2, 400 ml photosynthetic bacteria agent+3 600 ml nutrient solution; T3, 600 ml photosynthetic bacteria agent+ 3 400 ml nutrient solution; T4, 800 ml photosynthetic bacteria agent+3 200 ml nutrient solution; and T5, 1 000 ml photosynthetic bacteria agent+3 000 ml nutrient solu- tion) on the leaf number, fresh mass, root vigor, vitamin C content, soluble sugar content, soluble protein content and nitrate content of hydroponic Lactuca sativa L. were investigated. The results showed that the leaf number was increased by 62.66%, the fresh mass was increased by 139.7%, the root vigor was increased by 132.04%, the vitamin C content was increased by 18.34%, the soluble protein content was increased by 16.60%, the soluble sugar content was increased by 192.37%, and the nitrate content was reduced by 69.44% in the T3 group com- pared with those in the control group. The photosynthetic bacteria solution in the T3 group reduced the content of nitrate and improved the yield and quality of hydroponic L. sativa.展开更多
In this paper, the function of the iel gene from baculovirus Spodoptera exigua multiple nucleopolyhedrovirus (SeMNPV), belonging to group II nucleopolyhedrovirus, was studied in mammalian cells We amplified the SeMN...In this paper, the function of the iel gene from baculovirus Spodoptera exigua multiple nucleopolyhedrovirus (SeMNPV), belonging to group II nucleopolyhedrovirus, was studied in mammalian cells We amplified the SeMNPV iel gene and expressed it by fusing to the C terminal of enhanced GFP protein in HEK 293 cells. Confocal microscopy revealed that the IE1-GFP fusion protein was localized in the nucleus of the mammalian cells. The promoter sequences of AcMNPV gp64, SeMNPV F protein and Drosophila hsp70 were also analyzed, to further study the function of SeMNPV IE1. The results showed that, in the absence of the hr sequence, IE1 improved the expression of the F promoter but didn't influence the gp64 promoter significantly, but IE1 moderately stimulated the hsp70 promoter.展开更多
文摘Two new ent-kauranoids, tenuifolin A (3beta,6alpha, 15beta-trihydroxy-1alpha, 7beta-diacetoxy-11beta, 16beta-epoxy-ent-kaurane) (1) and tenuifolin B (1alpha,6alpha, 11beta-trihydroxy-3beta,7beta-diacetoxy-ent-kaur-16-en-15-one) (2), together with four known compounds were isolated from the aerial parts of Isodon tenuifolia (W. W. Smith) Kudo collected from Zhongdian County, Yunnan Province, China. Their structures were determined by the spectral methods (including 2D NMR techniques).
基金This work was supported by the National High Technology R&D Project of China (No.2002AA207009) and Wuhan Dawn Project for Youth (No. 20035002016-36).
文摘Northern blot analysis was conducted with mitochondrial RNA from seedling leaves, floral buds, and developing seeds of NCa CMS, maintainer line and fertile F1 using ten mitochondrial genes as probes. The results revealed that 9 out of the 10 mitochondrial genes, except for atp6, showed no difference in different tissues of the corresponding materials of NCα CMS system and that they might be constitutively expressed genes. Eight genes, such as orf139, orf222, atpl, cox1, cox2, cob, rm5S, and rm26S, showed no difference among the three tissues of all the materials detected. So the expression of these eight genes was not regulated by nuclear genes and was not tissue-specific. The transcripts of atp9 were identical among different tissues, but diverse among different materials, indicating that transcription of atp9 was neither controlled by nuclear gene nor tissue-specific. Gene atp6 displayed similar transcripts with the same size among different tissues of all the materials but differed in abundance among tissues of corresponding materials and its expression might be tissue-specific under regulation of nuclear gene. Moreover, three transcripts of orf222 were detected in the floral buds of NCa cms and fertile F1, but no transcript was detected in floral buds of the maintainer line.The transcription of orf139 was similar to that of orf222 but only two transcripts of 0.8 kb and 0.6 kb were produced. The atp9 probe detected a single transcript of 0.6 kb in NCa cms and in maintainer line and an additional transcript of 1.2 kb in fertile F1. The relationship of expression of orf222, orf139, and atp9 with NCa sterility was discussed.
文摘The ultrastructure of the vegetative cells of Nostoc flagelliforme Born. et Flah. was investigated with high pressure freezing and freeze substitution technique and compared with the results obtained by using conventional preparation methods. During the processes of chemical fixation, dehydration and embedding, the cell structures might be more artificially modified than that obtained from high pressure freezing and freeze substitution. With the present method, the sheath of N. flagelliforme could be well penetrated and no extra big space could exist between the cell and the sheath. The cell protoplasm rarely shrinked. Some fine structures of cell inclusions and unit membranes became visualized. Many bacteria were harbored in the sheath. In addition, the presence of big vacuoles in the cell of N. flagelliforme as well as the presence of bacteria in the sheath shown in the present preparation for cyanobacteria has not been described so far in the literature.
基金Supported by National Natural Science Foundation of China(Grant No.30960178)Natural Science Foundation of Xinjiang Province(Grant No.200821126)~~
文摘[Objective] This study aimed to investigate the feasibility of agricultural irri- gation with reclaimed water in Urumqi City. [Method] Chinese cabbages were used as experimental materials and irrigated with control water, 50% reclaimed water and 100% reclaimed water, and then the number of endophytic bacteria in Chinese cab- bages was measured. [Result] Using 50% reclaimed water, the fresh weight of Chi- nese cabbage was improved by 68.94%; however, with the deepening internalization of exogenous microorganism internalization, the total number of endophytic bacteria and coliform flock in Chinese cabbages treated with 50% reclaimed water was sig- nificantly higher than the other two treatment groups (P〈0.05). [Conclusion] Re- claimed water enhances both the weight of Chinese cabbage and the number of endophytic bacteria, which is unsuitable for irrigation of edible crops such as Chi- nese cabbage. Strengthening disinfection during the process of sewerage treatment could solve this problem.
文摘Cytochrome b_559 in photosystem Ⅱ reaction center was purified from spinach ( Spinacia oleracea L.) and rice ( Oryza sativa L.) by a rapid and simple procedure. Their low temperature fluorescence emission and excitation spectra, ultraviolet fluorescence spectra and absolute absorption spectra were presented. The author's purification methods, which enhanced the yield of pure protein and shorted the time for isolation, have several advantages: 1. use of oxygen_evolving PSⅡ core complexes as the starting material in order to avoid disturbing from other cytochromes; 2. isocratic elution of cytochrome b_559 from a DEAE_Sephacel column for eliminating the impurity and yielding the protein in pure state; 3. a simple column procedure for removal of excess Triton X_100. Purified cytochromes b_559 from these species have similar optical spectra and mobility during gel electrophoresis under native conditions. From the results of novel electrophoresis (Tricine_SDS_PAGE), cytochrome b_559 from both spinach and rice reveal two polypeptide bands (apparent molecular weight 9 kD and 4 kD, respectively). By measuring of 77 K fluorescence spectra, it was shown that for the purified cytochrome b_559 there were two excitation peaks at 439 nm and 413 nm, and two emission peaks at 563 nm and 668 nm. This is the first indication that Cyt b_559 is able to emit fluorescence and also transfer excited electrons to chlorophyll. By the use of ultraviolet fluorescence spectra, it was demonstrated for the first time that the location of Trp residue could be in the hydrophobic transmembrane region of cytochrome b_559.
基金Supported by National Natural Science Foundation of China (30471182)Youth Science Foundation of Hebei Province (C2010000738)~~
文摘Abstract [Objective] The paper was to prepare and detect the extended DNA fibers of Chinese cabbage. [Method] Chinese cabbage nuclei was first successfully isolated by chopping young leaves with a blade, then nuclei were lysed by SDS to release DNA, and DNA fibers were dragged and extended with a coverslip. [Result] The results of Fiber-FISH with genomic DNA and 25S rDNA as probes showed that DNA fiber size as long as about 1.93 Mb could be measured and the number of 25S rDNA copies region were estimated to be 258 and 687 in Chinese cabbage genome. DNA fibers prepared by this method showed equally spread parallel thread with clear background, and were suitable for FISH analysis. [Conclusion] The study would accelerate Chinese cabbage genome mapping and organization analysis.
基金Supported by "11thFive-Year" Crop Breeding Research of SichuanProvince "11thFive-Year" Joint Breeding Research Project Fun-ding of Sichuan Province.~~
文摘[Objective] The aim was to study the reproduction of the three-line genic male sterile (GMS) lineparent Mian7MB-1 (B. NapusL.) and the seed production of F1 through somatic tissue culture. [Methed] Through hybridization, a new breeding material Mian 7MB-1 in three-line genic temporary maintainer line propagated by tissue culture was used to improve the sterile plant rate of rapeseed in dual-purpose recessive GMS line, such as Mian 7AB type, S45AB type, and etc. And then the variety comparative test was performed. [Result] In order to avoid some fertility restoration phenomena occurring during the process of self-reproduction, Mian 7AB was propagated in bulk with somatic tissue culture of temporary maintainer line plant stem. The propagated temporary maintainer line seedlings were applied to the breeding and seed production of net room male sterile line parent, promoting the sterile plant rate of the male sterile line parent to 91.7% -93.5%. The male sterile line parents per hectare were enough for the seed production of hybrid F1 in 7 500 -15 000 hm^2. [ Conclusion ] Compared with the original dual-purpose GMS line, the seed production ultilizing male sterile line with high sterile plant rate greatly reduced the labor, significantly improved the seed yield, ensuring the seed quality and forming a perfect breeding and seed production system.
文摘It had been demonstrated that individual cells or protoplasts isolated fromPorphyrathallus by enzyme could develop into normal leafy thalli in the same way as monospores, and that isolated cells develop in different way in liquid and on semi-solid media. The authors observed the ultrastructure of isolated vegetative cells cultured on semi-solid media and compared them with those of monospores and isolated cells cultured in liquid media. The results showed that subcellular structures were quite different among cells in different conditions. In their development, isolated cells on semi-solid media did not show the characteristic subcellular feature of monospore formation, such as production of fibrous vesicles. Callus-like cells formed on semi-solid media underwent a distinctive modification in cellular organization. They developed characteristic cell inclusions and a special 2-layer cell covering. Golgi bodies, ER, starch grains, mitochondria. Vacuoles were not commonly found in them.
基金Supported by Bioreactor Important Special Item of 863-Program inthe "Eleventh Five-Year" Plan (No. 2007AA100503)Science and Technology Development Key Plan of Jilin Province( No.20070922)+1 种基金Cultivation Fund of Scientific and Technical Innovation Project Major Program of Higher Education Institutions ( No.70S018)Science and Technology Plan of Changchun City (No.06GG150)~~
文摘[ Objective] The study is to generate pharmaceutical protein via plant transgenic technique. [Methed] Using the cotyledons with petiole as transformation receptor, the fusion gene of rapeseed oil-body gene and bFGF was introduced into the rapeseed ( Brassica campestris L. ) by Agrobacterium tumefaciens-mediated transformation; meanwhile regeneration conditions of rapeseed were also optimized, and the regenerated resistant plantlets were detected by PCR and Southern blot. [ Result] This fusion gene had been integrated into rapeseed genome successfully, and the optimized conditions of transformation and regeneration were as follows: explants pre-culture for 2 d, co-culture for 3 d, bacteria solution OD600 for 0.3 and infection time for 5 min. [ Conclusion] The results laid a solid foundation for extraction, isolation and purification of protein in transgenic plant seeds.
文摘AIM: To investigate the potential effects of chelidonine, the main alkaloid of Chelidonium majus, on telomerase activity and its regulation in HepG2 cells. METHODS: Cytotoxicity of chelidonine for HepG2 cells was determined by neutral red assay. A modified polymerase chain reaction (PCR)-based telomerase repeat amplification protocol was used to estimate relative telomerase activity in chelidonine-treated cells in comparison with the untreated control cells. Relative expression level of the catalytic subunit of telomerase (hTERT) gene and P-glycoprotein (pgp) were estimated using semi-quantitative real-time reverse transcription-PCR (RT-PCR). Cell senescence in treated cells was demonstrated using a 13-galactosidase test. RESULTS: Cytotoxicity of chelidonine in HepG2 cells was not dose-dependent and tended to reach plateau immediately after the living cells were reduced in number to slightly higher than 50%. However, 12 μmol/L concentration of chelidonine was considered as LD50, where the maximal attainable effects were realized. Real-time RT-PCR data showed that the expression of pgp increased three-fold in chelidonine treated HepG2 cells in comparison with the untreated controls. MoP phologically, treated HepG2 cells showed apoptotic features after 24 h and a small fraction of cells appeared with single blister cell death. The relative expression level of Bcl-2 dropped to less than 50% of control cells at a sub-apoptotic concentration of chelidonine and subsequently increased to higher than 120% at LD50. Telomerase activity was reduced considerably after administration of very low doses of chelidonine, whereas higher concentrations of chelidonine did not remarkably enhance the effect. Real-time RT-PCR experiments indicated a drastic decrease in expression level of hTERT subunit of telomerase under treatment with chelidonine. Repeated treatment of cells with very low doses of chelidonine caused a decline in growth rate by 4 wk and many of the cells appeared to be aged with large volume and dark staining in the β-galactosidase assay. CONCLUSION: Chelidonine reduces telomerase activity through down-regulation of hTERT expression. Senescence induction might not be directly caused by reducing telomerase activity as it occurs after a few population doublings.
文摘AIM: To evaluate the antioxidant and phase II detoxification enzyme inducing ability of green leaf vegetables consumed in Asia.METHODS: The antioxidant properties of six commonly consumed Asian vegetables were determined using the ABTS, DPPH, deoxyribose, PR bleaching and ironascorbate induced lipid peroxidation assay. Induce of phase II detoxification enzymes was also determined for each respective vegetable extract. Protection against authentic ONOO- and HOCl mediated cytotoxicity in human colon HCT116 cells was determined using the MTT 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrasodium bromide) viability assay.RESULTS: All of the extracts derived from green leaf vegetables exhibited antioxidant properties, while also having cytoprotective effects against ONOO- and HOCI mediated cytotoxicity. In addition, evaluation of the phase II enzyme inducing ability of each extract, as assessed by quinone reductase and glutathione- S-transferase activities, showed significant variation between the vegetables analyzed.CONCLUSION: Green leaf vegetables are potential sources of antioxidants and phase II detoxification enzyme inducers in the Asian diet. It is likely that consumption of such vegetables is a major source of beneficial phytochemical constituents that may protect against colonic damage.
文摘The effects of different concentrations of photosynthetic bacteria solutions (CK, 0 ml photosynthetic bacteria agent+4 000 ml nutrient solution; T1, 200 ml pho- tosynthetic bacteria agent+3 800 ml nutrient solution; T2, 400 ml photosynthetic bacteria agent+3 600 ml nutrient solution; T3, 600 ml photosynthetic bacteria agent+ 3 400 ml nutrient solution; T4, 800 ml photosynthetic bacteria agent+3 200 ml nutrient solution; and T5, 1 000 ml photosynthetic bacteria agent+3 000 ml nutrient solu- tion) on the leaf number, fresh mass, root vigor, vitamin C content, soluble sugar content, soluble protein content and nitrate content of hydroponic Lactuca sativa L. were investigated. The results showed that the leaf number was increased by 62.66%, the fresh mass was increased by 139.7%, the root vigor was increased by 132.04%, the vitamin C content was increased by 18.34%, the soluble protein content was increased by 16.60%, the soluble sugar content was increased by 192.37%, and the nitrate content was reduced by 69.44% in the T3 group com- pared with those in the control group. The photosynthetic bacteria solution in the T3 group reduced the content of nitrate and improved the yield and quality of hydroponic L. sativa.
基金The knowledge innovation program of the Chinese Academy of Sciences (KSCX2-YW-Z-0938)
文摘In this paper, the function of the iel gene from baculovirus Spodoptera exigua multiple nucleopolyhedrovirus (SeMNPV), belonging to group II nucleopolyhedrovirus, was studied in mammalian cells We amplified the SeMNPV iel gene and expressed it by fusing to the C terminal of enhanced GFP protein in HEK 293 cells. Confocal microscopy revealed that the IE1-GFP fusion protein was localized in the nucleus of the mammalian cells. The promoter sequences of AcMNPV gp64, SeMNPV F protein and Drosophila hsp70 were also analyzed, to further study the function of SeMNPV IE1. The results showed that, in the absence of the hr sequence, IE1 improved the expression of the F promoter but didn't influence the gp64 promoter significantly, but IE1 moderately stimulated the hsp70 promoter.