背景:诱导骨髓基质干细胞成骨条件的优化与探讨是再生医学研究的热点。前期实验发现Wnt7b在骨发育中的作用,此次实验是前期工作的延续。目的:构建Wnt7b重组反转录病毒载体,观察其在C3H10T1/2中的表达。设计、时间及地点:细胞学体外观察,...背景:诱导骨髓基质干细胞成骨条件的优化与探讨是再生医学研究的热点。前期实验发现Wnt7b在骨发育中的作用,此次实验是前期工作的延续。目的:构建Wnt7b重组反转录病毒载体,观察其在C3H10T1/2中的表达。设计、时间及地点:细胞学体外观察,于2005-01/2008-08在华盛顿大学医学院和同济大学附属第十人民医院中心实验室完成。材料:pXy-Wnt7b购于ATCC公司,载体pCIG-IRES-EGFP,pLef-Luciferase,pCMV-Rennilla,反转录病毒载体pSFG和包装细胞GP293由华盛顿大学医学院提供。方法:用多步亚克隆技术构建目的基因Wnt7b和标记基因加强型绿色荧光蛋白(enhanced green fluorescent protein,EGFP)双表达重组反转录病毒载体pSFG-Wnt7b-IRES-EGFP,在条件培养液中经GP293包装,并在常规培养液中释放出假病毒并将其转染C3H10T1/2细胞,检测成骨活性诱导中的作用和荧光素酶报告基因检测信号转导通路。主要观察指标:①Wnt7b重组反转录病毒转染C3H10T1/2细胞的表达。②Wnt7b诱导C3H10T1/2碱性磷酸酶生成分析。③Wnt7b信号转导分析。结果:构建目的基因的Wnt7b和标记基因EGFP双表达重组反转录病毒载体能够高效地表达,转染C3H10T1/2细胞后能诱导碱性磷酸酶的生成。Wnt7b信号转导分析可见Wnt7b成骨作用通过Noncanonical途径。结论:Wnt7b在C3H10T1/2中能高效表达,能显著诱导骨发育的重要标志--碱性磷酸酶的生成,Noncanonical途径在其中起重要作用。展开更多
[ Objective] The study aimed to lay a foundation for the further studies on function mechanism of NS1 protein in the interspecies transmission of waterfowl influenza virus. [Method] Using the serologic assay and the s...[ Objective] The study aimed to lay a foundation for the further studies on function mechanism of NS1 protein in the interspecies transmission of waterfowl influenza virus. [Method] Using the serologic assay and the specific RT-PCR method, some strains of H9 subtype waterfowl influenza virus were isolated from the 12 to 20 day-old muscovy duck flocks without any clinical symptoms in different areas of Guangdong Province. Four of these strains, including A/duck/ZQ/303/2007（H9N2） （A3 for short）, A/Duck/FJ/301/2007 （H9N2） （C1 for short）, A/Duck/NH/306/2007（H9N2） （ D6 for short）, A/duck/SS/402/2007（H9N2） （ E2 for short）, and a strain named A/duck/ZC/2007（H9N2） （L1 for short） from a muscovy duck died of avian influenza virus （AIV）, were used for NSl gene cloning and sequencing. Subsequently, the obtained NSl gene sequences were compared with other NS1 sequences registered in GenBank, and the phylogenetic analysis was also conducted. [Result] When compared with the H9N2 AIV NS1 sequences in GenBank, the NSl genes of the four AIV strains A3, C1, 136 and E2 displayed homologies ranging from 99% to 100% at nucleotide level, and 95% to 100% at amino acid level; while the NSl gene of L1 strain displayed homology ranging from 94% to 97% at nucleotide level, and 93% to 98% at amino acid level. The phylogenetic tree demonstrated that A3, C1, D6 and E2 were highly resemblant, and L1 was closest to AY66473 （chicken, 2003）. By comparison with the NS1 gene sequences of L1, AF523514 （duck）, AY664743 （chicken） and EF155262.1 （quail） using DNAstar, A3, C1, D6 and E.2 presented nucleotide variations at site 21 （ R→Q）, 70, 71 （ KE→EG）, 86 （ A→S）, 124 （V→M） and 225 （ S→N）, and amino acid variations at site 21,70, 71 and 86 in dsRNA- dependent protein kinase （PKR） binding domain of NSl gene, which induced the evident variations of antigenic determinant and surface proba- bility plot of NS1 protein. [ Conclusion] This study suggested that the amino acid sequence variation in PKR binding domain of NS1 protein had something to do with the virus pathogenicity.展开更多
In this study, we investigated the in vitro antiviral activity of the mycelia of higher mushrooms against influenza virus type A(serotype H1N1) and herpes simplex virus type 2(HSV-2), strain BH. All 10 investigated mu...In this study, we investigated the in vitro antiviral activity of the mycelia of higher mushrooms against influenza virus type A(serotype H1N1) and herpes simplex virus type 2(HSV-2), strain BH. All 10 investigated mushroom species inhibited the reproduction of influenza virus strain A/FM/1/47(H1N1) in MDCK cells reducing the infectious titer by 2.0–6.0 lg ID50. Four species, Pleurotus ostreatus, Fomes fomentarius, Auriporia aurea, and Trametes versicolor, were also determined to be effective against HSV-2 strain BH in RK-13 cells, with similar levels of inhibition as for influenza. For some of the investigated mushroom species—Pleurotus eryngii, Lyophyllum shimeji, and Flammulina velutipes—this is the first report of an anti-influenza effect. This study also reports the first data on the medicinal properties of A. aurea, including anti-influenza and antiherpetic activities. T. versicolor 353 mycelium was found to have a high therapeutic index(324.67), and may be a promising material for the pharmaceutical industry as an anti-influenza and antiherpetic agent with low toxicity. Mycelia with antiviral activity were obtained in our investigation by bioconversion of agricultural wastes(amaranth flour after CO2 extraction), which would reduce the cost of the final product and solve some ecological problems.展开更多
Objective To explore metabolic changes after acute hypoxia and modulating effect of vitamins B1, B2, and PP supplementation in mice exposed to acute hypoxia. Methods Fifty male Kunming mice were randomly divided into ...Objective To explore metabolic changes after acute hypoxia and modulating effect of vitamins B1, B2, and PP supplementation in mice exposed to acute hypoxia. Methods Fifty male Kunming mice were randomly divided into 5 groups： normal, acute hypoxia, acute hypoxia with 2, 4 and 8 time- vitamins B1, B2, and PP supplementation . All mice were fed with corresponding diets for two weeks and then were exposed to a simulated altitude of 6 000 meters for 8 h, except for the normal group. Nuclear magnetic resonance analysis was used to identify the changes of serum metabolic profiles. Results There were significant changes in some serum metabolites under induced acute hypoxia, essentially relative increase in the concentrations of lactate, sugar and lipids and decrease in ethanol. The serum levels of choline, succinate, taurine, alanine, and glutamine also increased and phosphocholine decreased in the acute hypoxia group. After vitamins B1, B2, and PP supplementation, all these metabolic changes gradually recovered. Conclusion Significant changes in serum metabolic profile were observed by metabolomics in mice exposed to acute hypoxia, and vitamins B1, B2, and PP supplementation proved to be beneficial to improving some metabolic pathways. It is suggested that the dietary intakes of vitamins B1, B2, and PP should be increased under hypoxia condition.展开更多
文摘背景:诱导骨髓基质干细胞成骨条件的优化与探讨是再生医学研究的热点。前期实验发现Wnt7b在骨发育中的作用,此次实验是前期工作的延续。目的:构建Wnt7b重组反转录病毒载体,观察其在C3H10T1/2中的表达。设计、时间及地点:细胞学体外观察,于2005-01/2008-08在华盛顿大学医学院和同济大学附属第十人民医院中心实验室完成。材料:pXy-Wnt7b购于ATCC公司,载体pCIG-IRES-EGFP,pLef-Luciferase,pCMV-Rennilla,反转录病毒载体pSFG和包装细胞GP293由华盛顿大学医学院提供。方法:用多步亚克隆技术构建目的基因Wnt7b和标记基因加强型绿色荧光蛋白(enhanced green fluorescent protein,EGFP)双表达重组反转录病毒载体pSFG-Wnt7b-IRES-EGFP,在条件培养液中经GP293包装,并在常规培养液中释放出假病毒并将其转染C3H10T1/2细胞,检测成骨活性诱导中的作用和荧光素酶报告基因检测信号转导通路。主要观察指标:①Wnt7b重组反转录病毒转染C3H10T1/2细胞的表达。②Wnt7b诱导C3H10T1/2碱性磷酸酶生成分析。③Wnt7b信号转导分析。结果:构建目的基因的Wnt7b和标记基因EGFP双表达重组反转录病毒载体能够高效地表达,转染C3H10T1/2细胞后能诱导碱性磷酸酶的生成。Wnt7b信号转导分析可见Wnt7b成骨作用通过Noncanonical途径。结论:Wnt7b在C3H10T1/2中能高效表达,能显著诱导骨发育的重要标志--碱性磷酸酶的生成,Noncanonical途径在其中起重要作用。
基金Supported by Key Specific Program for Science and Technology of Guangdong Province (2008B020700003 A2007A020400006)~~
文摘[ Objective] The study aimed to lay a foundation for the further studies on function mechanism of NS1 protein in the interspecies transmission of waterfowl influenza virus. [Method] Using the serologic assay and the specific RT-PCR method, some strains of H9 subtype waterfowl influenza virus were isolated from the 12 to 20 day-old muscovy duck flocks without any clinical symptoms in different areas of Guangdong Province. Four of these strains, including A/duck/ZQ/303/2007（H9N2） （A3 for short）, A/Duck/FJ/301/2007 （H9N2） （C1 for short）, A/Duck/NH/306/2007（H9N2） （ D6 for short）, A/duck/SS/402/2007（H9N2） （ E2 for short）, and a strain named A/duck/ZC/2007（H9N2） （L1 for short） from a muscovy duck died of avian influenza virus （AIV）, were used for NSl gene cloning and sequencing. Subsequently, the obtained NSl gene sequences were compared with other NS1 sequences registered in GenBank, and the phylogenetic analysis was also conducted. [Result] When compared with the H9N2 AIV NS1 sequences in GenBank, the NSl genes of the four AIV strains A3, C1, 136 and E2 displayed homologies ranging from 99% to 100% at nucleotide level, and 95% to 100% at amino acid level; while the NSl gene of L1 strain displayed homology ranging from 94% to 97% at nucleotide level, and 93% to 98% at amino acid level. The phylogenetic tree demonstrated that A3, C1, D6 and E2 were highly resemblant, and L1 was closest to AY66473 （chicken, 2003）. By comparison with the NS1 gene sequences of L1, AF523514 （duck）, AY664743 （chicken） and EF155262.1 （quail） using DNAstar, A3, C1, D6 and E.2 presented nucleotide variations at site 21 （ R→Q）, 70, 71 （ KE→EG）, 86 （ A→S）, 124 （V→M） and 225 （ S→N）, and amino acid variations at site 21,70, 71 and 86 in dsRNA- dependent protein kinase （PKR） binding domain of NSl gene, which induced the evident variations of antigenic determinant and surface proba- bility plot of NS1 protein. [ Conclusion] This study suggested that the amino acid sequence variation in PKR binding domain of NS1 protein had something to do with the virus pathogenicity.
文摘In this study, we investigated the in vitro antiviral activity of the mycelia of higher mushrooms against influenza virus type A(serotype H1N1) and herpes simplex virus type 2(HSV-2), strain BH. All 10 investigated mushroom species inhibited the reproduction of influenza virus strain A/FM/1/47(H1N1) in MDCK cells reducing the infectious titer by 2.0–6.0 lg ID50. Four species, Pleurotus ostreatus, Fomes fomentarius, Auriporia aurea, and Trametes versicolor, were also determined to be effective against HSV-2 strain BH in RK-13 cells, with similar levels of inhibition as for influenza. For some of the investigated mushroom species—Pleurotus eryngii, Lyophyllum shimeji, and Flammulina velutipes—this is the first report of an anti-influenza effect. This study also reports the first data on the medicinal properties of A. aurea, including anti-influenza and antiherpetic activities. T. versicolor 353 mycelium was found to have a high therapeutic index(324.67), and may be a promising material for the pharmaceutical industry as an anti-influenza and antiherpetic agent with low toxicity. Mycelia with antiviral activity were obtained in our investigation by bioconversion of agricultural wastes(amaranth flour after CO2 extraction), which would reduce the cost of the final product and solve some ecological problems.
基金supported by a grant from the National Natural Science Foundation of China (NO. 30872097)
文摘Objective To explore metabolic changes after acute hypoxia and modulating effect of vitamins B1, B2, and PP supplementation in mice exposed to acute hypoxia. Methods Fifty male Kunming mice were randomly divided into 5 groups： normal, acute hypoxia, acute hypoxia with 2, 4 and 8 time- vitamins B1, B2, and PP supplementation . All mice were fed with corresponding diets for two weeks and then were exposed to a simulated altitude of 6 000 meters for 8 h, except for the normal group. Nuclear magnetic resonance analysis was used to identify the changes of serum metabolic profiles. Results There were significant changes in some serum metabolites under induced acute hypoxia, essentially relative increase in the concentrations of lactate, sugar and lipids and decrease in ethanol. The serum levels of choline, succinate, taurine, alanine, and glutamine also increased and phosphocholine decreased in the acute hypoxia group. After vitamins B1, B2, and PP supplementation, all these metabolic changes gradually recovered. Conclusion Significant changes in serum metabolic profile were observed by metabolomics in mice exposed to acute hypoxia, and vitamins B1, B2, and PP supplementation proved to be beneficial to improving some metabolic pathways. It is suggested that the dietary intakes of vitamins B1, B2, and PP should be increased under hypoxia condition.
