Objective] This study aimed to discuss the key technology of in-vitro cul-ture for a new Vaccinium uliginosum cultivar Zishuijing. [Method] Through the screen-ing and optimization of sterilization method for explants,...Objective] This study aimed to discuss the key technology of in-vitro cul-ture for a new Vaccinium uliginosum cultivar Zishuijing. [Method] Through the screen-ing and optimization of sterilization method for explants, sampling time, multiplication, nursing and rooting culture, a matching clone propagation system was established for the new Vaccinium uliginosum cultivar Zishuijing. [Result] The explants were sterilized with 0.1% HgCl2 for 3 min; the differentiation and multiplication medium of Zishuijing was composed of WPM （modified）, 6-BA （1.0 mg/L） and ZT （1.0 MG/L）; the rootless tube seedlings were transplanted in organic matrix （sawdust∶bark∶peat=1∶1∶1） in September and cultured at air relative humidity of 80%-90% and temperature of 20-25 ℃, and after 50 d, the rooting rate reached 72.4%. [Conclusion] The key technol-ogy of in-vitro culture for the new Vaccinium uliginosum cultivar Zishuijing was estab-lished, thereby providing technical support for large-scale industrialized seedling pro-duction of Zishuijing.展开更多
Background Vaccinium uliginosum L. is a type of blueberry found in the Chinese Changbai Mountains. We extracted Vaccinium uliginosum Anthocyanins （Av.uli） to investigate its bioactivity on suppressing cancer cells. ...Background Vaccinium uliginosum L. is a type of blueberry found in the Chinese Changbai Mountains. We extracted Vaccinium uliginosum Anthocyanins （Av.uli） to investigate its bioactivity on suppressing cancer cells. Methods Av.lli was extracted under different conditions of temperature （10℃-35℃）, pH 1.0-3.0, and diatomaceous earth （1.0 g-3.0 g）, followed by a HPLC analysis for the determination of the ingredients. Its anticancer bioactivities on human colon and colorectal cancer cells （DLD-1 and COLO205） were compared with those on Lonicera caerulea Anthocyanins （AL.cae） and Vaccinium myrtillus Anthocyanins （Av.myr）, using cell viability assays, DNA electrophoresis and nuclear morphology assays. Results The optimum process of Av.uli extraction involved conditions of temperature 20℃, pH 2.0, and diatomaceous earth 1.0 g/50 g of fruit weight. Av.uli contained 5 main components： delphinidin （40.70±1.72）%, cyanidin （3.40±0.68）%, petunidin （17.70±0.54）%, peonidin （2.90±0.63）% and malvidin （35.50±1.11）%. The malvidin percentage was significantly higher （P 〈0.05） than it in Av.myr. Av.uli complied with a dose-dependent repression of cancer cell proliferation with an IC50 （50% inhibitory concentration） value of 50 μg/ml, and showed greater anticancer efficiency than AL. cae and Av. myr under the same cell treatment conditions. These observations were further supported by the results of nuclear assays. Conclusions The extraction protocol and conditions we used were effective for anthocyanin extraction. Av.uli could be a feasible practical research tool and a promising therapeutic source to suppress human colon or colorectal cancers.展开更多
Objective： To study the effect of Vaccinium uliginosum L., （VU） on the electroretinogram （ERG） and retinal pathological changes in rabbits after light-induced damage. Methods： Twenty-eight Chinchilla rabbits wer...Objective： To study the effect of Vaccinium uliginosum L., （VU） on the electroretinogram （ERG） and retinal pathological changes in rabbits after light-induced damage. Methods： Twenty-eight Chinchilla rabbits were randomly divided into four groups： administration beforehand （A）, administration after injury （B）, light injury without administration （C）, and blank （D） groups. After a 4-week administration of VU homogenate at 4.8 g/（kg.d） once a day in group A, ERG in groups A, B and C were recorded according to the standards set by the International Society for Clinical Electrophysiology of Vision （ISCEV）. Except for group D, the groups were then exposed to strong light. Just after that, group A stopped receiving VU treatment and group B started to receive it. Then ERGs in all groups were recorded after 1 day, 1 week, and 2 weeks. Throughout the whole process groups which were not fed with VU were fed with normal saline. Finally, the tissues and structures of all the groups were observed and the thickness of the outer nuclear layers （ONL） was measured. Results： （1） After 4-week feeding with VU, the latency time of ERG in group A became shorter than those in the other groups and the amplitude increased. After being exposed to strong light, the latency time lengthened and amplitude decreased in all the injury groups, but comparing at each time point, the measured values in group A were better than those in group C. With the accumulation of VU, the ERG in group B improved, and finally, all of the detected values became better than those in group C. （2） Retinae in group D were normal in histology and the layers were in order but those in group C became disarranged. The injudes in groups A and B were minor compared with those in group C. The thickness of the ONL in group C was significantly thinner than in the other groups （all P=0.000）, and that in groups A and B was thicker than that in group C, although thinner than in group D. That in group A was thicker than in group B. Conclusions： VU can relieve the injury to rabbit retinae exposed to normal day and night rhythm, alleviate the harm caused by light when used beforehand, and repair the light damage to the retina.展开更多
Objective: To study the protective effect of anthocyanins extracted from Vaccinium Uliginosum(VU)on retinal 661W cells against microwave radiation induced retinal injury. Methods: 661W cells were divided into 6 groups...Objective: To study the protective effect of anthocyanins extracted from Vaccinium Uliginosum(VU)on retinal 661W cells against microwave radiation induced retinal injury. Methods: 661W cells were divided into 6 groups, including control, model [661W cells radiated by microwave(30 mW/cm2, 1 h)] and VU groups [661W cells pretreated with anthocyanins extracted from VU(25, 50, 100 and 200 μg/mL, respectively) for 48 h, and radiated by microwave 30 mW/cm2, 1 h]. After treatment with different interventions, the cell apoptosis index(AI)was determined using Heochst staining;contents of malonaldehyde(MDA), glutataione(GSH), and activity of superoxide dismutase(SOD) were measured. mRNA expressions of nuclear factor erythroid 2-related factor 2(Nrf2) and heme oxygenase 1(HO-1) were detected by real time quantitative polymerase chain reaction, and the expression of HO-1 protein was examined by Western blot analysis. Nucleus and cytoplasm were separated and Nrf2 protein expression was further verified by Western blot analysis. Results: There was significant difference in AI among the groups(F=322.83, P<0.05). Compared with the control group, AI was significantly higher in the model group and was lower in 4 VU-pretreated groups(P<0.05). Linear regression analysis showed the decline of AI was in a dose-dependent manner with VU treatment(r=0.8419, P<0.05). The MDA and GSH contents of 661W cells in VU-treated groups were significantly lower than the model group(P<0.05). Compared with the model group, the SOD activity in the VU-treated groups(50, 100 and 200 μg/mL) was significantly higher(all P<0.05). The Nrf2 and HO-1 mRNA expressions were slightly increased after irradiation, and obviously increased in 100 μg/mL VU-treated group. After irradiation, the relative expressions of HO-1 and Nrf2 proteins in nucleus were slightly increased(P<0.05), and the changes in cytoplasm were not obvious,whereas it was significantly increased in both nucleus and cytoplasm in the VU treatment groups. Conclusions:Anthocyanins extracted from VU could reduce apoptosis, stabilize cell membrane, and alleviate oxidant injury of mouse retinal photoreceptor 661W cells. The mechanism might be through activating Nrf2/HO-1 signal pathway and inducing HO-1 transcription and translation.展开更多
基金Supported by Key Project of the National Twelfth-Five Year Research Program of China(2011BAD08B01-03)Funding Project of Department of Forestry of Heilongjiang Province(201004068-6)~~
文摘Objective] This study aimed to discuss the key technology of in-vitro cul-ture for a new Vaccinium uliginosum cultivar Zishuijing. [Method] Through the screen-ing and optimization of sterilization method for explants, sampling time, multiplication, nursing and rooting culture, a matching clone propagation system was established for the new Vaccinium uliginosum cultivar Zishuijing. [Result] The explants were sterilized with 0.1% HgCl2 for 3 min; the differentiation and multiplication medium of Zishuijing was composed of WPM （modified）, 6-BA （1.0 mg/L） and ZT （1.0 MG/L）; the rootless tube seedlings were transplanted in organic matrix （sawdust∶bark∶peat=1∶1∶1） in September and cultured at air relative humidity of 80%-90% and temperature of 20-25 ℃, and after 50 d, the rooting rate reached 72.4%. [Conclusion] The key technol-ogy of in-vitro culture for the new Vaccinium uliginosum cultivar Zishuijing was estab-lished, thereby providing technical support for large-scale industrialized seedling pro-duction of Zishuijing.
文摘Background Vaccinium uliginosum L. is a type of blueberry found in the Chinese Changbai Mountains. We extracted Vaccinium uliginosum Anthocyanins （Av.uli） to investigate its bioactivity on suppressing cancer cells. Methods Av.lli was extracted under different conditions of temperature （10℃-35℃）, pH 1.0-3.0, and diatomaceous earth （1.0 g-3.0 g）, followed by a HPLC analysis for the determination of the ingredients. Its anticancer bioactivities on human colon and colorectal cancer cells （DLD-1 and COLO205） were compared with those on Lonicera caerulea Anthocyanins （AL.cae） and Vaccinium myrtillus Anthocyanins （Av.myr）, using cell viability assays, DNA electrophoresis and nuclear morphology assays. Results The optimum process of Av.uli extraction involved conditions of temperature 20℃, pH 2.0, and diatomaceous earth 1.0 g/50 g of fruit weight. Av.uli contained 5 main components： delphinidin （40.70±1.72）%, cyanidin （3.40±0.68）%, petunidin （17.70±0.54）%, peonidin （2.90±0.63）% and malvidin （35.50±1.11）%. The malvidin percentage was significantly higher （P 〈0.05） than it in Av.myr. Av.uli complied with a dose-dependent repression of cancer cell proliferation with an IC50 （50% inhibitory concentration） value of 50 μg/ml, and showed greater anticancer efficiency than AL. cae and Av. myr under the same cell treatment conditions. These observations were further supported by the results of nuclear assays. Conclusions The extraction protocol and conditions we used were effective for anthocyanin extraction. Av.uli could be a feasible practical research tool and a promising therapeutic source to suppress human colon or colorectal cancers.
文摘Objective： To study the effect of Vaccinium uliginosum L., （VU） on the electroretinogram （ERG） and retinal pathological changes in rabbits after light-induced damage. Methods： Twenty-eight Chinchilla rabbits were randomly divided into four groups： administration beforehand （A）, administration after injury （B）, light injury without administration （C）, and blank （D） groups. After a 4-week administration of VU homogenate at 4.8 g/（kg.d） once a day in group A, ERG in groups A, B and C were recorded according to the standards set by the International Society for Clinical Electrophysiology of Vision （ISCEV）. Except for group D, the groups were then exposed to strong light. Just after that, group A stopped receiving VU treatment and group B started to receive it. Then ERGs in all groups were recorded after 1 day, 1 week, and 2 weeks. Throughout the whole process groups which were not fed with VU were fed with normal saline. Finally, the tissues and structures of all the groups were observed and the thickness of the outer nuclear layers （ONL） was measured. Results： （1） After 4-week feeding with VU, the latency time of ERG in group A became shorter than those in the other groups and the amplitude increased. After being exposed to strong light, the latency time lengthened and amplitude decreased in all the injury groups, but comparing at each time point, the measured values in group A were better than those in group C. With the accumulation of VU, the ERG in group B improved, and finally, all of the detected values became better than those in group C. （2） Retinae in group D were normal in histology and the layers were in order but those in group C became disarranged. The injudes in groups A and B were minor compared with those in group C. The thickness of the ONL in group C was significantly thinner than in the other groups （all P=0.000）, and that in groups A and B was thicker than that in group C, although thinner than in group D. That in group A was thicker than in group B. Conclusions： VU can relieve the injury to rabbit retinae exposed to normal day and night rhythm, alleviate the harm caused by light when used beforehand, and repair the light damage to the retina.
文摘Objective: To study the protective effect of anthocyanins extracted from Vaccinium Uliginosum(VU)on retinal 661W cells against microwave radiation induced retinal injury. Methods: 661W cells were divided into 6 groups, including control, model [661W cells radiated by microwave(30 mW/cm2, 1 h)] and VU groups [661W cells pretreated with anthocyanins extracted from VU(25, 50, 100 and 200 μg/mL, respectively) for 48 h, and radiated by microwave 30 mW/cm2, 1 h]. After treatment with different interventions, the cell apoptosis index(AI)was determined using Heochst staining;contents of malonaldehyde(MDA), glutataione(GSH), and activity of superoxide dismutase(SOD) were measured. mRNA expressions of nuclear factor erythroid 2-related factor 2(Nrf2) and heme oxygenase 1(HO-1) were detected by real time quantitative polymerase chain reaction, and the expression of HO-1 protein was examined by Western blot analysis. Nucleus and cytoplasm were separated and Nrf2 protein expression was further verified by Western blot analysis. Results: There was significant difference in AI among the groups(F=322.83, P<0.05). Compared with the control group, AI was significantly higher in the model group and was lower in 4 VU-pretreated groups(P<0.05). Linear regression analysis showed the decline of AI was in a dose-dependent manner with VU treatment(r=0.8419, P<0.05). The MDA and GSH contents of 661W cells in VU-treated groups were significantly lower than the model group(P<0.05). Compared with the model group, the SOD activity in the VU-treated groups(50, 100 and 200 μg/mL) was significantly higher(all P<0.05). The Nrf2 and HO-1 mRNA expressions were slightly increased after irradiation, and obviously increased in 100 μg/mL VU-treated group. After irradiation, the relative expressions of HO-1 and Nrf2 proteins in nucleus were slightly increased(P<0.05), and the changes in cytoplasm were not obvious,whereas it was significantly increased in both nucleus and cytoplasm in the VU treatment groups. Conclusions:Anthocyanins extracted from VU could reduce apoptosis, stabilize cell membrane, and alleviate oxidant injury of mouse retinal photoreceptor 661W cells. The mechanism might be through activating Nrf2/HO-1 signal pathway and inducing HO-1 transcription and translation.
