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Aim To observe the effect of Gupi Xiaoji Decoction (GPXJY) on the structure and function of mitochondria of human hepatoma cell HepG2 cells and explore its possible mechanism. Methods CCK8 was used to detect cell proliferation, Mito-Tracker Green fluorescence staining was used to observe the mitochondrial structure, flow cytometry was used to detect the membrane potential, Elisa was used to detect the ATP content, fluoroscopic electron microscopy was used to observe the microstructure changes, and high-content screening(HCS) was used to detect the related proteins. Results Fluorescence staining showed that GPXJY damaged the mitochondria of HepG2 cells and decreased the content of ATP. The results of flow cytometry showed that GPXJY could reduce the mitochondrial membrane potential of HepG2 cells. The results of electron microscope showed that GPXJY made the mitochondria of cancer cells swell and so on. HCS found that GPXJY significantly reduced the average fluorescence intensity of Bcl-2 in HepG2 cells, and significantly increased the average fluorescence intensity of apoptosis promoting proteins Bax, cytochrome-c, caspase-3 and cleaved-caspase-3, which was statistically significant. Conclusion GPXJY can regulate the structure and function of mitochondria in HepG2 cells.
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ABSTRACT: The aim of this study was to study the effect of the auxin naphthalene acetic acid (NAA) and the brassinosteroid 28-homocastasterone (28-HCS) applied every 15 days (harvest 2015/16) or every 21 days (harvest 2016/17) after full bloom, on the physical, chemical and biochemical attributes of the 'Galaxy' apples. The study was performed at a commercial orchard at Vacaria county, Rio Grande do Sul State, Brazil. The following treatments were applied: water (control), NAA (0.1%), 28-HCS (10‒6 M) and NAA (0.1%) + 28-HCS (10‒6 M). For this purpose, 300 μL of the plant growth regulator solutions were applied to the fruit peduncles. Fruits treated with NAA every 21 days presented higher starch-iodine index and lower total soluble solids (TSS), titratable acidity (TA), lightness index (L) and hue angle (h°) than 28-HCS-treated fruits. Fruits treated every 15 days with NAA presented reduced skin rupture force (SRF), total antioxidant activities (TAA) of the skin and flesh, and hydrogen peroxide content compared to the control fruits. In addition, NAA application every 15 and 21 days resulted in increased anthocyanin content in the skin. Application of NAA + 28-HCS reduced superoxide dismutase activity. 28-HCS applications increased TSS, color attributes (C and h°) of the skin, total antioxidant activity of the skin and flesh and peroxidase activity compared to control fruits. These results are expected to help to understand how plant growth regulators affect apple quality. In addition, results described here are also expected to help on the development of strategies to reduce post harvest losses and to increase fruit shelf life.
RESUMO: O objetivo deste estudo foi avaliar a ação do ácido naftaleno acético (ANA) e do brassinosteroide (catasterona), aplicados a cada 15 (safra 2015/16) ou 21 (safra 2016/17) dias a partir dos 40 dias após a plena floração, nos atributos físico-químicos e bioquímicos de maçãs 'Galaxy'. O estudo foi realizado em um pomar comercial de macieiras 'Galaxy' no município de Vacaria, RS. Os tratamentos avaliados foram: água (controle), ANA (0,1%), catasterona (10‒6 M) e ANA (0,1%) + catasterona (10‒6 M). As aplicações foram realizadas no pedúnculo dos frutos, utilizando 300 µL de solução por pedúnculo. Frutos tratados com ANA a cada 21 dias tiveram maior índice de iodo-amido e menores valores de sólidos solúveis (SS), acidez titulável e dos atributos de cor L e ângulo hue (h°) do que os frutos tratados com catasterona. Aplicações de ANA a cada 15 dias reduziu a força de ruptura da casca (FRC), atividade antioxidante (AAT) da casca, da polpa e conteúdo de peróxido de hidrogênio. Além disso, ANA aplicada a cada 15 ou 21 dias aumentaram o teor de antocianinas na casca. ANA + catasterona reduziu a atividade da enzima superóxido dismutase. Entretanto, aplicações de catasterona a cada 15 dias aumentou o teor de SS, atributos de cor (C e h°) na casca, atividade antioxidante da casca e da polpa, compostos fenólicos totais (CFT) e atividade da enzima peroxidase dos frutos de maçãs. Portanto, estudos adicionais devem ser realizados para confirmar a ação desses fitorreguladores na manutenção da qualidade e redução das perdas pós-colheitas de maçãs 'Galaxy'.
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Objective To investigate the clinical, biochemical and genetic features of a Chinese boy with holocarboxylase synthetase deficiency (HCSD). Methods The clinical and genetic data of a rare case of HCSD were retrospectively analyzed. Results After birth, the boy showed development delay. At 3 months old, the boy was started with rehabilitation. Tandem mass spectrum and gas chromatography analysis was carried in the 5th month after birth because of the recurrent upper respiratory tract infection and elevated level of C5-OH in the blood and decreased level of C0,and elevated level of 3-OH-propionic, pyruvic acid, methylcrotonylglycine in the urine were in accordance with the HCSD. Genetic analysis found compound heterozygous mutations of c.1648G>A and c.1544G>A in gene, of which the latter one is novel. After the treatment of biotin (20 mg/d) and L-Carnitine, the condition of this boy was gradually improved. Conclutions HCSD is characterized with slow onset and inconspicuous manifestations. The confirmed diagnosis can be built with MS/MS, GC/MS analysis and gene mutation analysis. The effect of early biotin treatment is satisfactory. In this study,we carried out clinical and genetic diagnosis,which lays a solid foundation for prenatal diagnosis and early treatment.
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Aim To optimize the most effective compo-nent formula from the active ingredients of Salvia Milti-orrhiza and Panax Ginseng through the orthogonal de-sign method to resist breast cancer, and to reveal its antitumor mechanism in MCF-7 cells. Methods The human breast cancer cells MCF-7 were employed as the research object and the normal breast epithelial cells MCF-10A were used as control,optimizing the most ef-fective component formula from the active ingredients of Salvia Miltiorrhiza and Panax Ginseng by using CCK-8 assay and orthogonal design method; real-time cell a-nalysis was used to monitor the best combination formu-la on cell proliferation, and high content screening was used to detect the best combination drug on cell apop-tosis. Results The best combination of the salvianolic acids, saponins of Panax Ginseng and ginseng polysac-charides that were screened out were 5 , 10 , 5 mg · L-1 . Compared with control group, the treatment group had effective response inhibiting the proliferation on MCF-7 cells, but those effects were weaker on MCF-10A cells through real-time cell analysis. Ho-echst, Annexin V, PI staining fluorescence showed no significant difference ( P >0. 05 ) on MCF-10 A cells compared with the control group,but there was signifi-cant difference ( P <0. 01 ) on MCF-7 cells by HCS. Conclusions The most effective component formula from the active ingredients of Salvia Miltiorrhiza and Panax Ginseng have a strong inhibition of proliferation and induction of apoptosis to resist breast cancer with selection, and there is no significant difference in MCF-10A cells.
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Remarkable advances in cellular reprogramming have made it possible to investigate relevant cell populations derived from induced pluripotent stem cells ( iPSCs ) of patients. Be-cause many diseases have its specific genetic information, using the cells to convert into iPSCs can build up a set of genetic pro-file of diseases. The iPSCs which contain the genetic contribution of the donor can be expanded and differentiated into cells of the affected lineages to show aberrant phenotypes in culture. To date, over fifty such disease models have been reported, and while the field is young and hurdles remain, we can foresee the huge potential of it in drug screening. Recent studies using iP-SCs to model various neurogenetic disorders are summarized. Compared to the traditional methods, we analyze the future de-velopment of iPSC based disease models and its past application on high-throughput screening ( HTS) and high-content screening ( HCS) .
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High content screening(HCS) analysis is an advanced technique in the era of systematic biology,which can comprehensively reflect the changes of cell with live,dynamic,and multiparametric characters.It is a good tool to study the complex ingredients and mechanism of traditional Chinese medicine(TCM).Application of the technique would contribute to the discovery of effective ingredients,elucidation of mechanism,and development of TCM theory.