Exogenous alanyl-glutamine（Aln-Gln） was evaluated for its effects on growth performance, intestinal structure and function, antioxidant status and non-specific immunity of young carp（Cyprinus carpio L.）. Six diets...Exogenous alanyl-glutamine（Aln-Gln） was evaluated for its effects on growth performance, intestinal structure and function, antioxidant status and non-specific immunity of young carp（Cyprinus carpio L.）. Six diets supplemented with 0, 2.5, 5.0, 7.5, 10.0, or 15.0 g · kg-1 of Aln-Gln were fed to fish for 12 weeks. Supplementation with 7.5, 10.0, or 15.0 g · kg-1 of Aln-Gln significantly increased weight gain rate（WGR）, protein efficiency ratio（PER）, but feed conservation rate（FCR） and survival were not affected（P〉0.05）. The intestinal fold height and number, digestive enzyme, Na＋, K＋-ATPase activities was found to be significantly high（P〈0.05） with increasing dietary Aln-Gln supplementation up to 7.5 g · kg-1, but there were no significant differences for Aln-Gln supplementation from 7.5 to 15.0 g · kg-1. The glutathione peroxidase（GPX） activity, glutathione（GSH）, superoxide dismutase（SOD） activity increased and malondialdehyde（MDA） levels decreased significantly（P〈0.05） in the intestine, hepatopancreas, plasma and muscles. The plasma complement-3（C3） and complement-4（C4） levels were significantly（P〈0.05） improved at 5.0 g · kg-1 level and decreased when over 7.5 g · kg-1. The plasma lysozyme（LSZ） activity increased significantly（P〈0.05） at 7.5, 10.0, or 15.0 g · kg-1 level. In summary, the results showed that Aln-Gln improved growth performance, development and function of the intestine, the activity of the antioxidant defense system and the plasma non-specific immunity of the carps. The optimal Aln-Gln level was 8.24 g · kg-1 diet for WGR based on broken-line regression model analysis.展开更多
The patterns of cleavage of mtDNA by restriction endonucleases was analysed for six fowl breeds and of. Hyline White, Hyline Brown, ISA Brown, Hisex Brown, Lohmann White, Abor Acres and mtDNA polymorphisms were detect...The patterns of cleavage of mtDNA by restriction endonucleases was analysed for six fowl breeds and of. Hyline White, Hyline Brown, ISA Brown, Hisex Brown, Lohmann White, Abor Acres and mtDNA polymorphisms were detected in the restriction patterns with the following eight enzymes, Ava Ⅰ, Ava Ⅱ, EcoR Ⅰ, Hind Ⅲ, Bam HI, Pvu Ⅱ, Pst Ⅰ, Hinc Ⅱ. The restriction cleavage patterns were identical among these breeds. Hyline White, Hyline Brown, ISA Brown, Hisex Brown, Lohmann White—A type. The patterns of Abor Acres were B type. Based on their mtDNA restriction types, all the breeds were classified into two groups. Genetic distances among these groups were calculated in order to define their phylogenetic relationships. The relationship among five egg line breeds is close, while Abor Acres (Broiler fowl) is relatively far from them. The results suggest that the difference of mtDNA could result from the different origins. The polymorphic sites in mtDNA of Hyline White has been located on a restriction map.展开更多
目的探讨减数分裂内切酶1(EME1)在肝癌组织中的表达及其对肝癌细胞生物学行为的影响。方法筛选TCGA数据库肝癌样本中的差异表达基因。采用免疫组化和Western Blot分析EME1在肝癌组织中的表达丰度。通过短发夹RNA(shRNA)构建慢病毒并感染...目的探讨减数分裂内切酶1(EME1)在肝癌组织中的表达及其对肝癌细胞生物学行为的影响。方法筛选TCGA数据库肝癌样本中的差异表达基因。采用免疫组化和Western Blot分析EME1在肝癌组织中的表达丰度。通过短发夹RNA(shRNA)构建慢病毒并感染BEL-7404细胞干扰EME1基因表达,分为沉默组(shEME1)和对照组(shCtrl)。通过实时荧光定量PCR法和Western Blot检测两组EME1 mRNA和蛋白表达水平,Celigo计数法及MTT活性检测细胞增殖率,流式细胞术检测细胞周期,Caspase3/7活性检测细胞凋亡。两组间比较采用成组t检验。结果TCGA结果显示EME1的mRNA表达水平在肝癌组织中是癌旁组织的18.9倍(114.5±153.0 vs 8.0±7.2,t=5.00,P<0.001);EME1的蛋白表达水平在肝癌组织中是癌旁组织的7.0倍(免疫组化检测,8.4±2.6 vs 1.2±0.4,t=7.55,P<0.001)和2.5倍(Western Blot检测,249.0%±35.5%vs 100.0%±77.8%,t=3.02,P<0.05)。慢病毒感染后,相对于对照组,沉默组EME1的mRNA表达水平下降了29.9%(29.9%±0.9%vs 100.0%±3.6%,t=32.82,P<0.001),蛋白表达水平显著下降了35.7%(35.7%±14.9%vs 100.0%±28.9%,t=3.42,P<0.05);细胞计数下降了45.1%(4053±167 vs 8988±477,t=16.91,P<0.001)、细胞活性下降至66.9%(0.518±0.046 vs 0.774±0.022,t=8.74,P<0.001)及细胞克隆形成能力下降至29.0%(75±6 vs 260±9,t=28.92,P<0.001)。与对照组比较,沉默组G1期细胞(49.9%vs 44.0%,t=8.96,P<0.001)比例增多,G2/M期(15.9%vs 17.9%,t=9.13,P<0.001)与S期(34.2%vs 38.1%,t=6.91,P<0.001)的细胞比例减少;Caspase3/7活性增强了1.5倍(145.8%±5.9%vs 100.0%±2.3%,t=12.50,P<0.001)。结论EME1在肝癌组织中高表达,沉默EME1基因可抑制肝癌细胞增殖,促进细胞凋亡。展开更多
目的探讨姜黄素(curcumin)对乳腺癌细胞中顺铂(Cisplatin,CDDP)治疗敏感性的影响及其可能机制。方法 CCK-8检测、Hoechst染色观察CDDP、姜黄素单独及联合用药48 h对MCF7细胞增殖抑制和细胞凋亡的影响;Western blot分析MCF7细胞在CDDP(0...目的探讨姜黄素(curcumin)对乳腺癌细胞中顺铂(Cisplatin,CDDP)治疗敏感性的影响及其可能机制。方法 CCK-8检测、Hoechst染色观察CDDP、姜黄素单独及联合用药48 h对MCF7细胞增殖抑制和细胞凋亡的影响;Western blot分析MCF7细胞在CDDP(0、1.25、2.5、5、10、20μg/m L)、姜黄素(0、1、5、20、30、50、100μmol/L)单独及联合处理后FEN1(flap endonuclease 1)表达水平的变化;CCK-8检测沉默FEN1对MCF7细胞中CDDP敏感性的影响。结果 CDDP、姜黄素均可以剂量依赖方式抑制MCF7细胞增殖,其IC50分别为5、30μmol/L;与单独2μg/m L CDDP处理组比较,2μg/m L CDDP联合20μmol/L姜黄素、30μmol/L姜黄素处理组对细胞增殖的抑制效应明显增强[分别为(84.1±0.8)%、(51.1±0.5)%、(29.4±0.3)%,P<0.05];与单独20μmol/L姜黄素处理组比较,20μmol/L姜黄素联合2μg/m L CDDP、5μg/m L CDDP处理组对细胞增殖的抑制效应也明显增强[分别为(76.9±0.7)%、(42.4±0.3)%、(31.6±0.4)%,P<0.05];2μg/m L CDDP联合20μmol/L姜黄素组的细胞凋亡明显增强(P<0.05);与Control siRNA组比较,FEN1 siRNA+CDDP组可显著增强CDDP抑制MCF7细胞增殖的敏感性[(25.4±0.3)%vs(18.7±0.2)%,P<0.05];CDDP增殖抑制无效浓度或低浓度时,FEN1蛋白的表达随CDDP的处理剂量依赖性上调,而姜黄素可剂量依赖性下调FEN1蛋白表达;与单独CDDP和姜黄素处理组比较,2μg/m L CDDP联合20μmol/L姜黄素组可显著降低FEN1蛋白表达(P<0.05)。结论姜黄素可增强CDDP对乳腺癌细胞的敏感性,其机制与降低细胞FEN1的表达有关。展开更多
基金Supported by the Earmarked Fund for China Agriculture Research System(CARS-46)the Special Scientific Research Funds for Central Non-profit Institutes,Chinese Academy of Fishery Sciences(2014A08XK03)
文摘Exogenous alanyl-glutamine（Aln-Gln） was evaluated for its effects on growth performance, intestinal structure and function, antioxidant status and non-specific immunity of young carp（Cyprinus carpio L.）. Six diets supplemented with 0, 2.5, 5.0, 7.5, 10.0, or 15.0 g · kg-1 of Aln-Gln were fed to fish for 12 weeks. Supplementation with 7.5, 10.0, or 15.0 g · kg-1 of Aln-Gln significantly increased weight gain rate（WGR）, protein efficiency ratio（PER）, but feed conservation rate（FCR） and survival were not affected（P〉0.05）. The intestinal fold height and number, digestive enzyme, Na＋, K＋-ATPase activities was found to be significantly high（P〈0.05） with increasing dietary Aln-Gln supplementation up to 7.5 g · kg-1, but there were no significant differences for Aln-Gln supplementation from 7.5 to 15.0 g · kg-1. The glutathione peroxidase（GPX） activity, glutathione（GSH）, superoxide dismutase（SOD） activity increased and malondialdehyde（MDA） levels decreased significantly（P〈0.05） in the intestine, hepatopancreas, plasma and muscles. The plasma complement-3（C3） and complement-4（C4） levels were significantly（P〈0.05） improved at 5.0 g · kg-1 level and decreased when over 7.5 g · kg-1. The plasma lysozyme（LSZ） activity increased significantly（P〈0.05） at 7.5, 10.0, or 15.0 g · kg-1 level. In summary, the results showed that Aln-Gln improved growth performance, development and function of the intestine, the activity of the antioxidant defense system and the plasma non-specific immunity of the carps. The optimal Aln-Gln level was 8.24 g · kg-1 diet for WGR based on broken-line regression model analysis.
文摘The patterns of cleavage of mtDNA by restriction endonucleases was analysed for six fowl breeds and of. Hyline White, Hyline Brown, ISA Brown, Hisex Brown, Lohmann White, Abor Acres and mtDNA polymorphisms were detected in the restriction patterns with the following eight enzymes, Ava Ⅰ, Ava Ⅱ, EcoR Ⅰ, Hind Ⅲ, Bam HI, Pvu Ⅱ, Pst Ⅰ, Hinc Ⅱ. The restriction cleavage patterns were identical among these breeds. Hyline White, Hyline Brown, ISA Brown, Hisex Brown, Lohmann White—A type. The patterns of Abor Acres were B type. Based on their mtDNA restriction types, all the breeds were classified into two groups. Genetic distances among these groups were calculated in order to define their phylogenetic relationships. The relationship among five egg line breeds is close, while Abor Acres (Broiler fowl) is relatively far from them. The results suggest that the difference of mtDNA could result from the different origins. The polymorphic sites in mtDNA of Hyline White has been located on a restriction map.
文摘目的探讨减数分裂内切酶1(EME1)在肝癌组织中的表达及其对肝癌细胞生物学行为的影响。方法筛选TCGA数据库肝癌样本中的差异表达基因。采用免疫组化和Western Blot分析EME1在肝癌组织中的表达丰度。通过短发夹RNA(shRNA)构建慢病毒并感染BEL-7404细胞干扰EME1基因表达,分为沉默组(shEME1)和对照组(shCtrl)。通过实时荧光定量PCR法和Western Blot检测两组EME1 mRNA和蛋白表达水平,Celigo计数法及MTT活性检测细胞增殖率,流式细胞术检测细胞周期,Caspase3/7活性检测细胞凋亡。两组间比较采用成组t检验。结果TCGA结果显示EME1的mRNA表达水平在肝癌组织中是癌旁组织的18.9倍(114.5±153.0 vs 8.0±7.2,t=5.00,P<0.001);EME1的蛋白表达水平在肝癌组织中是癌旁组织的7.0倍(免疫组化检测,8.4±2.6 vs 1.2±0.4,t=7.55,P<0.001)和2.5倍(Western Blot检测,249.0%±35.5%vs 100.0%±77.8%,t=3.02,P<0.05)。慢病毒感染后,相对于对照组,沉默组EME1的mRNA表达水平下降了29.9%(29.9%±0.9%vs 100.0%±3.6%,t=32.82,P<0.001),蛋白表达水平显著下降了35.7%(35.7%±14.9%vs 100.0%±28.9%,t=3.42,P<0.05);细胞计数下降了45.1%(4053±167 vs 8988±477,t=16.91,P<0.001)、细胞活性下降至66.9%(0.518±0.046 vs 0.774±0.022,t=8.74,P<0.001)及细胞克隆形成能力下降至29.0%(75±6 vs 260±9,t=28.92,P<0.001)。与对照组比较,沉默组G1期细胞(49.9%vs 44.0%,t=8.96,P<0.001)比例增多,G2/M期(15.9%vs 17.9%,t=9.13,P<0.001)与S期(34.2%vs 38.1%,t=6.91,P<0.001)的细胞比例减少;Caspase3/7活性增强了1.5倍(145.8%±5.9%vs 100.0%±2.3%,t=12.50,P<0.001)。结论EME1在肝癌组织中高表达,沉默EME1基因可抑制肝癌细胞增殖,促进细胞凋亡。
文摘目的探讨姜黄素(curcumin)对乳腺癌细胞中顺铂(Cisplatin,CDDP)治疗敏感性的影响及其可能机制。方法 CCK-8检测、Hoechst染色观察CDDP、姜黄素单独及联合用药48 h对MCF7细胞增殖抑制和细胞凋亡的影响;Western blot分析MCF7细胞在CDDP(0、1.25、2.5、5、10、20μg/m L)、姜黄素(0、1、5、20、30、50、100μmol/L)单独及联合处理后FEN1(flap endonuclease 1)表达水平的变化;CCK-8检测沉默FEN1对MCF7细胞中CDDP敏感性的影响。结果 CDDP、姜黄素均可以剂量依赖方式抑制MCF7细胞增殖,其IC50分别为5、30μmol/L;与单独2μg/m L CDDP处理组比较,2μg/m L CDDP联合20μmol/L姜黄素、30μmol/L姜黄素处理组对细胞增殖的抑制效应明显增强[分别为(84.1±0.8)%、(51.1±0.5)%、(29.4±0.3)%,P<0.05];与单独20μmol/L姜黄素处理组比较,20μmol/L姜黄素联合2μg/m L CDDP、5μg/m L CDDP处理组对细胞增殖的抑制效应也明显增强[分别为(76.9±0.7)%、(42.4±0.3)%、(31.6±0.4)%,P<0.05];2μg/m L CDDP联合20μmol/L姜黄素组的细胞凋亡明显增强(P<0.05);与Control siRNA组比较,FEN1 siRNA+CDDP组可显著增强CDDP抑制MCF7细胞增殖的敏感性[(25.4±0.3)%vs(18.7±0.2)%,P<0.05];CDDP增殖抑制无效浓度或低浓度时,FEN1蛋白的表达随CDDP的处理剂量依赖性上调,而姜黄素可剂量依赖性下调FEN1蛋白表达;与单独CDDP和姜黄素处理组比较,2μg/m L CDDP联合20μmol/L姜黄素组可显著降低FEN1蛋白表达(P<0.05)。结论姜黄素可增强CDDP对乳腺癌细胞的敏感性,其机制与降低细胞FEN1的表达有关。
