Objective:To investigate the suitability of citrus-press cakes,by-products of the juice industry as a source for the whitening agents for cosmetic industry.Methods:Ethylacetate extracts of citrus-press cakes(CCE)were ...Objective:To investigate the suitability of citrus-press cakes,by-products of the juice industry as a source for the whitening agents for cosmetic industry.Methods:Ethylacetate extracts of citrus-press cakes(CCE)were examined for their anti-melanogenic potentials in terms of the inhibition of melanin production and mechanisim of melanogenesis by using Western Blot analysis with tyrosinese,tyrosinase-related protein-1(TRP-1),TRP2,and microphthalmia-associated transcription factor(MITF)proteins.To apply the topical agents,citrus-press cakes was investigated the safety in human skin cell line.Finally flavonoid analysis of CCE was also determined by HPLC analysis.Results:Results indicated that CCE were shown to down-regulate melanin content in a dose-dependent pattern.The CCE inhibited tyrosinase,TRP-2,and MITF expressions in a dose-dependent manner.To test the applicability of CCE to human skin,we used MTT assay to assess the cytotoxic effects of CCE on human keratinocyte HaCaT cells.The CCE exhibited low cytotoxicity at 50μg/mL.Characterization of the citrus-press cakes for flavonoid contents using HPLC showed varied quantity of rutin,narirutin,and hesperidin.Conclusions:Considering the anti-melanogenic activity and human safety,CCE is considered as a potential anti-melanogenic agent and may be effective for topical application for treating hyperpigmentation disorders.展开更多
miRNA是在许多生物过程中都起着至关重要作用的一类内源性非编码的小RNA,与癌症、肿瘤的发生有关。现发现很多miRNA在黑色素生成中都有重要的调控作用,但miR-146a是否对黑色素的生成具有影响未见报道。本研究发现miR-146a通过靶向抑制...miRNA是在许多生物过程中都起着至关重要作用的一类内源性非编码的小RNA,与癌症、肿瘤的发生有关。现发现很多miRNA在黑色素生成中都有重要的调控作用,但miR-146a是否对黑色素的生成具有影响未见报道。本研究发现miR-146a通过靶向抑制酪氨酸酶相关蛋白1(tyrosinase related protein 1,TYRP1)的表达而使黑色素生成降低。在小鼠黑色素细胞中分别转染miR-146a mimic和miR-146a抑制剂,通过qRT-PCR与Western印迹分析比较各实验组中TYRP1基因与酪氨酸家族相关基因酪氨酸酶(tyrosinase,TYR)、酪氨酸酶相关蛋白2(tyrosinase related protein 2,TYRP2)的表达差异。双荧光报告实验验证TYRP1与miR-146a的靶向关系,双荧光酶活性结果显示,实验组相比对照组,荧光素酶活性明显降低,说明TYRP1是miR-146a的靶基因之一;qRT-PCR和Western印迹结果显示实验组TYR、TYRP1及TYRP2在mRNA水平和蛋白质水平表达均显著降低;紫外分光光度法检测黑色素含量,结果显示miR-146a mimic转染组黑色素含量明显下降,而抑制组的黑色素含量呈上升趋势。综上所述,miR-146a通过靶向抑制TYRP1基因的表达,而影响TYR家族成员的表达,调控黑色素的生物合成。展开更多
AIM:To investigate the effect of the G-1666A polymorphism in the multidrug resistance related protein-1 (MRP1) on outcome of hepatocellular carcinoma (HCC). METHODS:A cohort of 162 patients with surgically resected HC...AIM:To investigate the effect of the G-1666A polymorphism in the multidrug resistance related protein-1 (MRP1) on outcome of hepatocellular carcinoma (HCC). METHODS:A cohort of 162 patients with surgically resected HCC who received no postsurgical treatment until relapse was studied. Genotyping was performed by polymerase chain reaction-restriction fragment length polymorphism analysis. Electrophoretic mobility shift assay (EMSA) was used to evaluate the influence of the G-1666A polymorphism on the binding affinity of the MRP1 promoter with its putative transcription factors. RESULTS:Kaplan-Meier analysis showed that patients with GG homologues had a reduced 4-year disease-free survival compared with those carrying at least one A allele (P = 0.011). Multivariate Cox regression analysis indicated that the-1666GG genotype represented an independent predictor of poorer disease-free survival [hazard ratio (HR) = 3.067,95% confidence interval (CI):1.587-5.952,P = 0.001],and this trend became worse in men (HR = 3.154,95% CI:1.604-6.201,P = 0.001). A similar association was also observed between 4-year overall survival and the polymorphism in men (HR = 3.342,95% CI:1.474-7.576,P = 0.004). Moreover,EMSA suggested that the G allele had a stronger binding affinity to nuclear proteins. CONCLUSION:The MRP1-1666GG genotype predicted a worse outcome and was an independent predictor of poor survival in patients with HCC from Southeast China.展开更多
Objective: To elucidate distinct functions of a recently identified cancer metastasis-associated molecule, related to testes-specific, vespid, and pathogenesis protein-1 (RTVP-1) in the mammalian immune system. Method...Objective: To elucidate distinct functions of a recently identified cancer metastasis-associated molecule, related to testes-specific, vespid, and pathogenesis protein-1 (RTVP-1) in the mammalian immune system. Methods: Immunohistochemical assays and functional analysis on the immune system were performed on RTVP-/- mice and RTVP-1+/+ mice. Protein-protein interaction (PPI) was used to predict the functions of RTVP-1. Results: Abnormal lymphocyte growth kinetics and reduced numbers of CD8+ T cells were revealed in lymph nodes of RTVP-1-/- mice. Expression of phenotypic markers of maturation in bone marrow-derived dendritic cells (DC) was impaired in RTVP-1-/- DC following antigenic stimulation in vitro and RTVP-1-/- DC failed to provide normal CD4+ T cell stimulatory activities in vivo. RTVP-1-/- mice failed to generate normal CTL or antibody responses in vivo after vaccination. In vivo tumor challenge experiments using a mouse cancer cell line demonstrated that the growth rate of subcutaneous tumors in RTVP-1-/- mice was significantly increased and CD8+ T cell infiltration significantly reduced compared to RTVP-1+/+ mice. PPI showed that RTVP-1 protein closely interacted with molecules associated with immune response and cancer metastasis. Conclusion: RTVP-1 might function as a tumor metastasis suppressor and immunosurveillance molecule in cancer.展开更多
目的:研究不同浓度小柴胡汤对B16黑素瘤细胞黑素合成相关酶酪氨酸酶(tyrosinase,TYR),酪氨酸酶相关蛋白-1(tyrosinase related protein-1,TRP-1),酪氨酸酶相关蛋白-2(tyrosinase related protein-2,TRP-2)mRNA表达的影响。方法:采用CCK...目的:研究不同浓度小柴胡汤对B16黑素瘤细胞黑素合成相关酶酪氨酸酶(tyrosinase,TYR),酪氨酸酶相关蛋白-1(tyrosinase related protein-1,TRP-1),酪氨酸酶相关蛋白-2(tyrosinase related protein-2,TRP-2)mRNA表达的影响。方法:采用CCK8检测细胞活力,选取小柴胡汤低、中、高质量浓度分别为0.1,1,2.5 g·L-1用于实验,逆转录-聚合酶链式反应(RTPCR)法检测B16细胞中TYR,TRP-1,TRP-2 mRNA的表达。结果:与正常组比较,随着浓度的增加,小柴胡汤水煎液能够显著下调细胞TYR,TRP-1,TRP-2 mRNA的表达(P<0.05,P<0.01)。结论:小柴胡汤对黑素合成的抑制作用除了通过直接抑制TYR活性外,还可通过下调酪氨酸酶家族其他成员TRP-1,TRP-2的基因表达、蛋白合成3方面实现其抑制黑素生成的作用,提示小柴胡汤对黑素生成的抑制可通过对酶蛋白转录后调节途径来实现。展开更多
基金Supported by the Next-Generation BioGreen 21 Program,Rural Development Administration,Republic of Korea,Grant No.PJ009583002013
文摘Objective:To investigate the suitability of citrus-press cakes,by-products of the juice industry as a source for the whitening agents for cosmetic industry.Methods:Ethylacetate extracts of citrus-press cakes(CCE)were examined for their anti-melanogenic potentials in terms of the inhibition of melanin production and mechanisim of melanogenesis by using Western Blot analysis with tyrosinese,tyrosinase-related protein-1(TRP-1),TRP2,and microphthalmia-associated transcription factor(MITF)proteins.To apply the topical agents,citrus-press cakes was investigated the safety in human skin cell line.Finally flavonoid analysis of CCE was also determined by HPLC analysis.Results:Results indicated that CCE were shown to down-regulate melanin content in a dose-dependent pattern.The CCE inhibited tyrosinase,TRP-2,and MITF expressions in a dose-dependent manner.To test the applicability of CCE to human skin,we used MTT assay to assess the cytotoxic effects of CCE on human keratinocyte HaCaT cells.The CCE exhibited low cytotoxicity at 50μg/mL.Characterization of the citrus-press cakes for flavonoid contents using HPLC showed varied quantity of rutin,narirutin,and hesperidin.Conclusions:Considering the anti-melanogenic activity and human safety,CCE is considered as a potential anti-melanogenic agent and may be effective for topical application for treating hyperpigmentation disorders.
文摘miRNA是在许多生物过程中都起着至关重要作用的一类内源性非编码的小RNA,与癌症、肿瘤的发生有关。现发现很多miRNA在黑色素生成中都有重要的调控作用,但miR-146a是否对黑色素的生成具有影响未见报道。本研究发现miR-146a通过靶向抑制酪氨酸酶相关蛋白1(tyrosinase related protein 1,TYRP1)的表达而使黑色素生成降低。在小鼠黑色素细胞中分别转染miR-146a mimic和miR-146a抑制剂,通过qRT-PCR与Western印迹分析比较各实验组中TYRP1基因与酪氨酸家族相关基因酪氨酸酶(tyrosinase,TYR)、酪氨酸酶相关蛋白2(tyrosinase related protein 2,TYRP2)的表达差异。双荧光报告实验验证TYRP1与miR-146a的靶向关系,双荧光酶活性结果显示,实验组相比对照组,荧光素酶活性明显降低,说明TYRP1是miR-146a的靶基因之一;qRT-PCR和Western印迹结果显示实验组TYR、TYRP1及TYRP2在mRNA水平和蛋白质水平表达均显著降低;紫外分光光度法检测黑色素含量,结果显示miR-146a mimic转染组黑色素含量明显下降,而抑制组的黑色素含量呈上升趋势。综上所述,miR-146a通过靶向抑制TYRP1基因的表达,而影响TYR家族成员的表达,调控黑色素的生物合成。
基金Supported by The Scientific and Technological Program of Guangdong Province, China, No. 2003B30102
文摘AIM:To investigate the effect of the G-1666A polymorphism in the multidrug resistance related protein-1 (MRP1) on outcome of hepatocellular carcinoma (HCC). METHODS:A cohort of 162 patients with surgically resected HCC who received no postsurgical treatment until relapse was studied. Genotyping was performed by polymerase chain reaction-restriction fragment length polymorphism analysis. Electrophoretic mobility shift assay (EMSA) was used to evaluate the influence of the G-1666A polymorphism on the binding affinity of the MRP1 promoter with its putative transcription factors. RESULTS:Kaplan-Meier analysis showed that patients with GG homologues had a reduced 4-year disease-free survival compared with those carrying at least one A allele (P = 0.011). Multivariate Cox regression analysis indicated that the-1666GG genotype represented an independent predictor of poorer disease-free survival [hazard ratio (HR) = 3.067,95% confidence interval (CI):1.587-5.952,P = 0.001],and this trend became worse in men (HR = 3.154,95% CI:1.604-6.201,P = 0.001). A similar association was also observed between 4-year overall survival and the polymorphism in men (HR = 3.342,95% CI:1.474-7.576,P = 0.004). Moreover,EMSA suggested that the G allele had a stronger binding affinity to nuclear proteins. CONCLUSION:The MRP1-1666GG genotype predicted a worse outcome and was an independent predictor of poor survival in patients with HCC from Southeast China.
基金the Grants 2003-406 from the Ministry of Education, China, and R01-50588 from the NCI, USA
文摘Objective: To elucidate distinct functions of a recently identified cancer metastasis-associated molecule, related to testes-specific, vespid, and pathogenesis protein-1 (RTVP-1) in the mammalian immune system. Methods: Immunohistochemical assays and functional analysis on the immune system were performed on RTVP-/- mice and RTVP-1+/+ mice. Protein-protein interaction (PPI) was used to predict the functions of RTVP-1. Results: Abnormal lymphocyte growth kinetics and reduced numbers of CD8+ T cells were revealed in lymph nodes of RTVP-1-/- mice. Expression of phenotypic markers of maturation in bone marrow-derived dendritic cells (DC) was impaired in RTVP-1-/- DC following antigenic stimulation in vitro and RTVP-1-/- DC failed to provide normal CD4+ T cell stimulatory activities in vivo. RTVP-1-/- mice failed to generate normal CTL or antibody responses in vivo after vaccination. In vivo tumor challenge experiments using a mouse cancer cell line demonstrated that the growth rate of subcutaneous tumors in RTVP-1-/- mice was significantly increased and CD8+ T cell infiltration significantly reduced compared to RTVP-1+/+ mice. PPI showed that RTVP-1 protein closely interacted with molecules associated with immune response and cancer metastasis. Conclusion: RTVP-1 might function as a tumor metastasis suppressor and immunosurveillance molecule in cancer.
文摘目的:研究不同浓度小柴胡汤对B16黑素瘤细胞黑素合成相关酶酪氨酸酶(tyrosinase,TYR),酪氨酸酶相关蛋白-1(tyrosinase related protein-1,TRP-1),酪氨酸酶相关蛋白-2(tyrosinase related protein-2,TRP-2)mRNA表达的影响。方法:采用CCK8检测细胞活力,选取小柴胡汤低、中、高质量浓度分别为0.1,1,2.5 g·L-1用于实验,逆转录-聚合酶链式反应(RTPCR)法检测B16细胞中TYR,TRP-1,TRP-2 mRNA的表达。结果:与正常组比较,随着浓度的增加,小柴胡汤水煎液能够显著下调细胞TYR,TRP-1,TRP-2 mRNA的表达(P<0.05,P<0.01)。结论:小柴胡汤对黑素合成的抑制作用除了通过直接抑制TYR活性外,还可通过下调酪氨酸酶家族其他成员TRP-1,TRP-2的基因表达、蛋白合成3方面实现其抑制黑素生成的作用,提示小柴胡汤对黑素生成的抑制可通过对酶蛋白转录后调节途径来实现。
