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1.
Nurs Health Sci ; 26(1): e13109, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38467127

RESUMO

Postmenopausal women with negative personality characteristics are at an increased risk of psychological disorders, yet little is known about the mechanism underlying the relationship between type D personality and psychological distress in postmenopausal women with coronary disease. This study assessed the mediating roles of perceived social support and self-perceived burden in the relationship between type D and psychological distress based on the equity theory and stress-buffering model. Demographic characteristics, type D, psychological distress, perceived social support, and self-perceived burden were completed by 335 participants with self-reported questionnaires using a cross-sectional design in Southeast China. The results revealed that perceived social support and self-perceived burden both separately and serially mediated the relationship between type D personality and psychological distress. Effective intervention strategies aimed at improving perceived social support or reducing self-perceived burden may be beneficial in reducing psychological distress.


Assuntos
Doença da Artéria Coronariana , Angústia Psicológica , Humanos , Feminino , Estudos Transversais , Análise de Mediação , Pós-Menopausa , Personalidade , Estresse Psicológico/complicações , Estresse Psicológico/psicologia , Apoio Social
2.
Environ Sci Pollut Res Int ; 30(54): 115295-115309, 2023 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-37880399

RESUMO

This study sought to examine hitherto unresearched relationships between serum terpenes and the prevalence of dyslipidemia. Serum terpenes such as limonene, α-pinene, and ß-pinene from the 2013-2014 National Health and Nutrition Examination Survey (NHANES) were used as independent variables in this cross-sectional study. Continuous lipid variables included total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), non-HDL-C, triglycerides (TG), low-density lipoprotein cholesterol (LDL-C), residual cholesterol (RC), and apolipoprotein B (Apo B). Binary lipid variables (elevated TC, ≥5.18 mmol/L; lowered HDL-C, <1.04 mmol/L in men, and <1.30 mmol/L in women; elevated non-HDL-C, ≥4.2 mmol/L; elevated TG, ≥1.7 mmol/L; elevated LDL-C, ≥3.37 mmol/L; elevated RC, ≥1.0 mmol/L; and elevated Apo B, ≥1.3 g/L) suggest dyslipidemia. The relationships between the mixture of serum terpenes with lipid variables were investigated using weighted quantile sum (WQS) regression and Bayesian kernel machine regression (BKMR). The study for TC, HDL-C, and non-HDL-C included a total of 1,528 people, whereas the analysis for TG, LDL-C, RC, and Apo B comprised 714 participants. The mean age of the overall participants was 47.69 years, and 48.77% were male. We found that tertiles of serum terpene were positively associated with binary (elevated TC, non-HDL-C, TG, LDL-C, RC, Apo B, and lowered HDL-C) and continuous (TC, non-HDL-C, TG, LDL-C, RC, and Apo B, but not HDL-C) serum lipid variables. WQS regression and BKMR analysis revealed that the mixture of serum terpenes was linked with the prevalence of dyslipidemia. According to our data, the prevalence of dyslipidemia was correlated with serum concentrations of three terpenes both separately and collectively.


Assuntos
Dislipidemias , Hipercolesterolemia , Humanos , Masculino , Feminino , Pessoa de Meia-Idade , LDL-Colesterol , Inquéritos Nutricionais , Terpenos , Prevalência , Estudos Transversais , Teorema de Bayes , Colesterol , Triglicerídeos , HDL-Colesterol , Dislipidemias/epidemiologia , Lipoproteínas , Apolipoproteínas B
3.
Hortic Res ; 10(9): uhad158, 2023 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-37719277

RESUMO

Although auxin is known to induce ethylene biosynthesis in some Rosaceae fruit crops, the mechanisms underlying the auxin-ethylene interaction during fruit ripening remain largely unknown. Here, the regulatory role of an auxin response factor, PpARF6, in fruit ripening was investigated in peach. Peach fruits showed accelerated ripening after treatment with auxin and PpARF6 was found to be significantly induced. PpARF6 not only could induce ethylene synthesis by directly activating the transcription of ethylene biosynthetic genes, but also competed with EIN3-binding F-box proteins PpEBF1/2 for binding to ethylene-insensitive3-like proteins PpEIL2/3, thereby keeping PpEIL2/3 active. Moreover, PpARF6 showed an interaction with PpEIL2/3 to enhance the PpEIL2/3-activated transcription of ethylene biosynthetic genes. Additionally, ectopic overexpression of PpARF6 in tomato accelerated fruit ripening by promoting the expression of genes involved in ethylene synthesis and fruit texture. In summary, our results revealed a positive regulatory role of PpARF6 in peach fruit ripening via integrating auxin and ethylene signaling.

4.
J Cardiovasc Nurs ; 2023 Aug 28.
Artigo em Inglês | MEDLINE | ID: mdl-37639559

RESUMO

BACKGROUND: Return to work (RTW) is a critical component of rehabilitation for most young and middle-aged patients after an acute myocardial infarction (AMI). Its success is related to the quality of life and social psychological function of patients, and their social economic growth. However, healthcare professionals often do not deeply understand the patients' experience and their difficulties and coping methods during this process, which limits their ability to institute effective management and support. OBJECTIVE: In this study, we aimed to explore the lived experiences and change processes of young and middle-aged patients with AMI at the different stages of RTW. METHODS: A descriptive qualitative approach was used. Patients aged 20 to 59 years with AMI were recruited from the Department of Cardiology of 3 general hospitals. Data were collected via semistructured interviews. Data analysis was performed by conventional content analysis methods. RESULTS: In total, 18 participants were included. Five main themes emerged: (1) "chaos," (2) "rebuilding," (3) "conflict," (4) "coping," and (5) "benefits." Patients may be more concerned about physical recovery during the initial clinical event. They then begin to plan and adjust for an RTW. Patients in the maintenance phase need strategies to prevent, identify, and respond to conflicts and challenges to maintain long-term stable work. CONCLUSION: We identified several post-AMI stages spanning from the initial illness event to the maintenance of stable work. We described their perceived barriers, coping strategies, and support needs at these various stages. These data are crucial for healthcare professionals to develop improved vocational rehabilitation strategies for patients with AMI.

5.
Environ Sci Pollut Res Int ; 30(29): 74290-74300, 2023 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-37204573

RESUMO

The relationship between aldehyde exposure and metabolic syndrome is unclear; hence, we aimed to investigate the association between serum aldehyde concentrations and metabolic syndrome. We analyzed the data of 1471 participants from the National Health and Nutrition Examination Survey enrolled from 2013 to 2014. The association of serum aldehyde concentrations with metabolic syndrome was assessed via generalized linear models as well as restricted cubic splines, and endpoint events were further analyzed. After adjusting for covariates, both moderate (odds ratio [OR] = 2.73, 95% confidence interval [CI]: 1.34-5.56) and high (OR = 2.08, 95% CI: 1.06-4.07) concentrations of isovaleraldehyde were associated with the risk of metabolic syndrome. Interestingly, although a moderate concentration of valeraldehyde was associated with the risk of metabolic syndrome (OR = 1.08, 95% CI: 0.70-1.65), a high concentration was not (OR = 0.55, 95% CI: 0.17-1.79). Restricted cubic splines revealed a non-linear association between valeraldehyde and metabolic syndrome, and threshold effect analysis revealed that the inflection point for valeraldehyde concentration was 0.7 ng/mL. The results of the subgroup analysis revealed differences in the relationship of aldehyde exposure with components of metabolic syndrome. High isovaleraldehyde concentrations may increase the risk of metabolic syndrome, and valeraldehyde demonstrated a J-shaped relationship with the risk of metabolic syndrome.


Assuntos
Síndrome Metabólica , Adulto , Humanos , Síndrome Metabólica/epidemiologia , Inquéritos Nutricionais , Aldeídos
6.
Hortic Res ; 9: uhac132, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35937864

RESUMO

Plant tissues are capable of developing unorganized cell masses termed calluses in response to the appropriate combination of auxin and cytokinin. Revealing the potential epigenetic mechanisms involved in callus development can improve our understanding of the regeneration process of plant cells, which will be beneficial for overcoming regeneration recalcitrance in peach. In this study, we report on single-base resolution mapping of DNA methylation and reprogramming of the pattern of trimethylation of histone H3 at lysine 27 (H3K27me3) at the genome-wide level during the leaf-to-callus transition in peach. Overall, mCG and mCHH were predominant at the genome-wide level and mCG was predominant in genic regions. H3K27me3 deposition was mainly detected in the gene body and at the TSS site, and GAGA repetitive sequences were prone to recruit H3K27me3 modification. H3K27me3 methylation was negatively correlated with gene expression. In vitro culture of leaf explants was accompanied by DNA hypomethylation and H3K27me3 demethylation, which could activate auxin- and cytokinin-related regulators to induce callus development. The DNA methylation inhibitor 5-azacytidine could significantly increase callus development, while the H3K27me3 demethylase inhibitor GSK-J4 dramatically reduced callus development. These results demonstrate the roles of DNA methylation and H3K27me3 modification in mediating chromatin status during callus development. Our study provides new insights into the epigenetic mechanisms through which differentiated cells acquire proliferative competence to induce callus development in plants.

7.
Tree Physiol ; 42(8): 1662-1677, 2022 08 06.
Artigo em Inglês | MEDLINE | ID: mdl-35220436

RESUMO

Peach Prunus persica is an economically important fruit tree crop worldwide. Although the external color of fruit is an important aspect of fruit quality, the mechanisms underlying its formation remain elusive in peach. Here, we report an elongated hypocotyl 5-homolog gene PpHYH involved in the regulation of anthocyanin pigmentation in peach fruit peel. Anthocyanin accumulation in fruit peel is light-dependent in peach. PpHYH had no auto-activation activity and its transcription was induced by sunlight. PpHYH activated transcription of a cluster of three PpMYB10 genes in the present of a cofactor PpBBX4 encoding a B-BOX protein, leading to anthocyanin accumulation in the sun-exposed peel. However, the PpHYH activity was repressed by a negative regulator of PpCOP1 encoding constitutive photomorphogenesis protein 1 which accumulated in the nucleus under dark condition, resulting in failure of anthocyanin accumulation in the shaded peel. PpCOP1 was re-localized into the cytosol under light condition, in accordance with fruit peel pigmentation. Additionally, transport of anthocyanins from the cytoplasm to the vacuole was a rate-limiting step for anthocyanin accumulation in peach fruit peel. Our results reveal for the first time the HYH gene involved in the regulation of anthocyanin accumulation in fruits, and provide target genes for genetic manipulation of fruit coloration.


Assuntos
Antocianinas , Prunus persica , Antocianinas/metabolismo , Frutas/genética , Frutas/metabolismo , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Prunus persica/genética , Prunus persica/metabolismo , Fatores de Transcrição/genética
8.
New Phytol ; 234(1): 179-196, 2022 04.
Artigo em Inglês | MEDLINE | ID: mdl-35023174

RESUMO

The fruit surface has an enormous impact on the external appearance and postharvest shelf-life of fruit. Here, we report two functionally redundant genes, PpMYB25 and PpMYB26, involved in regulation of fruit skin texture in peach. PpMYB25 can activate transcription of PpMYB26 and they both induce trichome development and cuticular wax accumulation, resulting in peach fruit with a fuzzy and dull appearance. By contrast, nonfunctional mutation of PpMYB25 caused by an insertional retrotransposon in the last exon in nectarine fails to activate transcription of PpMYB26, resulting in nectarine fruit with a smooth and shiny appearance due to loss of trichome initiation and decreased cuticular wax accumulation. Secondary cell wall biosynthesis in peach fruit pubescence is controlled by a transcriptional regulatory network, including the master regulator PpNAC43 and its downstream MYB transcription factors such as PpMYB42, PpMYB46 and PpMYB83. Our results show that PpMYB25 and PpMYB26 coordinately regulate fruit pubescence and cuticular wax accumulation and their simultaneous perturbation results in the origin of nectarine, which is botanically classified as a subspecies of peach.


Assuntos
Prunus persica , Frutas , Regulação da Expressão Gênica de Plantas , Genes myb , Prunus persica/genética , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Tricomas/genética , Tricomas/metabolismo
9.
Physiol Plant ; 173(4): 2119-2129, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34537956

RESUMO

Double flower is an invaluable trait in ornamental peach, but the mechanism underlying its development remains largely unknown. Here, we report the roles of ABCE model genes in double flower development in peach. A total of nine ABCE regulatory genes, including eight MADS-box genes and one AP2/EREBP gene, were identified in the peach genome. Subcellular localization assay showed that all the ABCE proteins were localized in the nucleus. Four genes, PpAP1, PpAP3, PpSEP3, and PpPI, showed a difference in expression levels between single and double flowers. Ectopic overexpression of PpPI increased petal number in Arabidopsis, while transgenic lines overexpressing PpAP3 or PpSEP3 were morphologically similar to wild-type. Ectopic overexpression of PpAP1 resulted in a significant decrease in the number of basal leaves and caused early flowering. These results suggest that PpPI is likely crucial for double flower development in peach. In addition, double flowers have petaloid sepals and stamens, and single flower could occasionally change to be double flower by converting stamens to petals in peach, suggesting that the double-flower trait is likely to have evolved from an ancestral single-flower structure. Our results provide new insights into mechanisms underlying the double-flower trait in peach.


Assuntos
Proteínas de Domínio MADS , Prunus persica , Flores/genética , Flores/metabolismo , Regulação da Expressão Gênica de Plantas/genética , Proteínas de Domínio MADS/genética , Proteínas de Domínio MADS/metabolismo , Filogenia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Prunus persica/genética , Prunus persica/metabolismo
10.
Front Pharmacol ; 12: 648107, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33935747

RESUMO

In this paper, a curcumin derivative Cur20 was synthesized for better hydrolytic stability, which showed a higher angiogenic effect on zebrafish model than curcumin. In order to reveal the potential effects on neuroprotection, a mouse model of vascular dementia (VaD) induced by permanent right common carotid artery occlusion (rUCCAO) was established. After two weeks of curcumin administration, the cognitive function of mice was detected by Morris water maze and Y maze. The alteration on oxidative injuries and morphological damage were also analyzed by reactive oxygen species, superoxide dismutase, GSH, malondialdehyde tests, and Nissl stain on cortex/hippocampus. The angiogenesis and related signal factors were evaluated as well. The results showed that Cur20 significantly attenuated the cognitive dysfunction and histopathological changes of the VaD mice with enhanced antioxidant system and angiogenesis. In addition, primary rat brain microvessel endothelial cells (rBMECs) with oxygen glucose deprivation (OGD) were applied to further verify the possible mechanisms of Cur20-induced angiogenesis. The results demonstrated that the proliferation effect and the activation of pro-angiogenesis factors such as HIF-1α, VEGF, and TFEB might contribute to the protection of ischemic injury. Based on the above, our conclusion is that Cur20 can be considered as a promising therapeutic strategy for VaD.

11.
Front Plant Sci ; 12: 653256, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33936139

RESUMO

Male sterility is an important agronomic trait for hybrid vigor utilization and hybrid seed production, but its underlying mechanisms remain to be uncovered. Here, we investigated the mechanisms of male sterility in peach using a combined cytology, physiology, and molecular approach. Cytological features of male sterility include deformed microspores and tapetum cells along with absence of pollen grains. Microspores had smaller nucleus at the mononuclear stage and were compressed into belts and subsequently disappeared in the anther cavity, whereas tapetum cells were swollen and vacuolated, with a delayed degradation to flowering time. Male sterile anthers had an ROS burst and lower levels of major antioxidants, which may cause abnormal development of microspores and tapetum, leading to male sterility in peach. In addition, the male sterility appears to be cytoplasmic in peach, which could be due to sequence variation in the mitochondrial genome. Our results are helpful for further investigation of the genetic mechanisms underlying male sterility in peach.

12.
Mol Plant ; 14(9): 1454-1471, 2021 09 06.
Artigo em Inglês | MEDLINE | ID: mdl-34022440

RESUMO

Although taste is an important aspect of fruit quality, an understanding of its genetic control remains elusive in apple and other fruit crops. In this study, we conducted genomic sequence analysis of 497 Malus accessions and revealed erosion of genetic diversity caused by apple breeding and possible independent domestication events of dessert and cider apples. Signatures of selection for fruit acidity and size, but not for fruit sugar content, were detected during the processes of both domestication and improvement. Furthermore, we found that single mutations in major genes affecting fruit taste, including Ma1, MdTDT, and MdSOT2, dramatically decrease malate, citrate, and sorbitol accumulation, respectively, and correspond to important domestication events. Interestingly, Ma1 was identified to have pleiotropic effects on both organic acid content and sugar:acid ratio, suggesting that it plays a vital role in determining fruit taste. Fruit taste is unlikely to have been negatively affected by linkage drag associated with selection for larger fruit that resulted from the pyramiding of multiple genes with minor effects on fruit size. Collectively, our study provides new insights into the genetic basis of fruit quality and its evolutionary roadmap during apple domestication, pinpointing several candidate genes for genetic manipulation of fruit taste in apple.


Assuntos
Frutas/genética , Malatos/metabolismo , Malus/genética , Mutação , Paladar , Evolução Biológica , Domesticação , Genes de Plantas/genética
13.
Front Public Health ; 9: 601791, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33996707

RESUMO

Objective: To assess the psychiatric status of Chinese civil servants aiding in coronavirus disease 2019 (COVID-19) control. Background: During the COVID-19 pandemic, Chinese civil servants have faced high workloads that may contribute to mental disorders. We assessed the prevalence of both depression and anxiety symptoms among civil servants in Jiangsu and surrounding provinces using the Chinese versions of the 9-item Patient Health Questionnaire (PHQ-9) and the 7-item Generalized Anxiety Disorder (GAD-7) scale. Methods: The PHQ-9 and GAD-7 were used to assess the severity of symptoms of depression and anxiety, respectively. Multivariable logistic regression analysis was performed to identify factors associated with mental health outcomes. Results: In total, 867 Chinese civil servants aiding in COVID-19 control were included in our study. Overall, 37.25 and 38.06% of all respondents reported having symptoms of depression and anxiety, respectively. Respondents who were younger and more educated and those who had fewer years of work experience had higher scores for both depression and anxiety. Multivariable logistic regression analysis showed that being a woman, being younger, having more education and having fewer years of work experience were associated with a higher risk of symptoms of depression and anxiety. However, whether they had experience combating infectious diseases or worked in frontline, there was no significant difference between respondents with and without experience, as well as between frontline and non-frontline workers, in both symptoms of depression and anxiety. Conclusions: The civil servants aiding in COVID-19 control reported suffering from varying degrees of mental disorders. Therefore, more attention should be devoted to the psychological distress of these civil servants.


Assuntos
COVID-19 , Pandemias , Estudos Transversais , Depressão/epidemiologia , Feminino , Humanos , Saúde Mental , Avaliação de Resultados em Cuidados de Saúde , SARS-CoV-2
14.
Food Chem ; 334: 127567, 2021 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-32707362

RESUMO

Fruit acidity is an important determinant of peach organoleptic quality, but its regulatory mechanism remains elusive. Measurement of organic acids in ripe fruits of seventy-five peach cultivars revealed the predominant components malate and citrate, accompanied by quinate. Organic acid accumulation increased at early stages of fruit growth, but exhibited a more dramatic reduction in low-acid cultivar during later stages of fruit development compared to high-acid cultivars. Low-acid cultivars showed citrate degradation and less transport of malate into the vacuole due to up- and down-regulation of a GABA pathway gene GAD and a malate transporter gene ALMT9, respectively. The NAD-MDH1 gene might control the rate-limiting step in malate synthesis, while three genes, PDK, PK, and ADH, could affect citrate synthesis through the pyruvate-to-acetyl-CoA-to-citrate pathway. Altogether, these results suggested that malate accumulation is controlled at the level of metabolism and vacuolar storage, while metabolism is crucial for citrate accumulation in peach.


Assuntos
Ácidos Carboxílicos/metabolismo , Frutas/metabolismo , Malatos/metabolismo , Proteínas de Plantas/genética , Prunus persica/metabolismo , Acetilcoenzima A/metabolismo , Ácido Cítrico/metabolismo , Frutas/genética , Frutas/crescimento & desenvolvimento , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/metabolismo , Prunus persica/genética , Prunus persica/crescimento & desenvolvimento , Ácido Pirúvico/metabolismo , Vacúolos/metabolismo
15.
BMC Plant Biol ; 20(1): 191, 2020 May 06.
Artigo em Inglês | MEDLINE | ID: mdl-32375636

RESUMO

BACKGROUND: Sugar content is an important determinant of fruit sweetness, but details on the complex molecular mechanism underlying fruit sugar accumulation remain scarce. Here, we report the role of sucrose transporter (SUT) family in regulating fruit sugar accumulation in apple. RESULTS: Gene-tagged markers were developed to conduct candidate gene-based association study, and an SUT4 member MdSUT4.1 was found to be significantly associated with fruit sugar accumulation. MdSUT4.1 encodes a tonoplast localized protein and its expression level had a negative correlation with fruit sugar content. Overexpression of MdSUT4.1 in strawberry and apple callus had an overall negative impact on sugar accumulation, suggesting that it functions to remobilize sugar out of the vacuole. In addition, MdSUT4.1 is located on chromosomal region harboring a previously reported QTL for sugar content, suggesting that it is a candidate gene for fruit sugar accumulation in apple. CONCLUSIONS: MdSUT4.1 is involved in the regulation of fruit sugar accumulation in apple. This study is not only helpful for understanding the complex mechanism of fruit sugar accumulation, but it also provides molecular tools for genetic improvement of fruit quality in breeding programs of apple.


Assuntos
Malus/metabolismo , Proteínas de Membrana Transportadoras/metabolismo , Proteínas de Plantas/metabolismo , Sacarose/metabolismo , Fragaria/genética , Frutas/genética , Estudos de Associação Genética , Genoma de Planta , Proteínas de Membrana Transportadoras/genética , Proteínas de Plantas/genética
16.
Sci Rep ; 10(1): 2836, 2020 02 18.
Artigo em Inglês | MEDLINE | ID: mdl-32071340

RESUMO

Peach is an economically import fruit crop worldwide, and serves as a model species of the Rosaceae family as well. However, peach functional genomics studies are severely hampered due to its recalcitrance to regeneration and stable transformation. Here, we report a fast and efficient Agrobacterium rhizogenes-mediated transformation system in peach. Various explants, including leaf, hypocotyl and shoot, were all able to induce transgenic hairy roots, with a transformation efficiency of over 50% for hypocotyl. Composite plants were generated by infecting shoots with A. rhizogenes to induce transgenic adventitious hairy roots. The composite plant system was successfully used to validate function of an anthocyanin-related regulatory gene PpMYB10.1 in transgenic hairy roots, and two downstream genes, PpUFGT and PpGST, were strongly activated. Our stable and reproductive A. rhizogenes-mediated transformation system provides an avenue for gene function assay, genetic engineering, and investigation of root-rhizosphere microorganism interaction in peach.


Assuntos
Genoma de Planta/genética , Genômica , Plantas Geneticamente Modificadas/genética , Prunus persica/genética , Agrobacterium/genética , Antocianinas/genética , Antocianinas/metabolismo , Engenharia Genética/tendências , Folhas de Planta/genética , Folhas de Planta/crescimento & desenvolvimento , Raízes de Plantas/genética , Raízes de Plantas/crescimento & desenvolvimento , Prunus persica/crescimento & desenvolvimento
17.
Reprod Domest Anim ; 54(2): 138-149, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30098220

RESUMO

In recent years, induced pluripotent stem cells (iPSCs) technique is able to allow us to generate pluripotency from somatic cells in vitro through the over expression of several transcription factors. Normally, viral vectors and transcription factors are commonly used on iPSC technique, which could cause many barriers on further application. In this study, we attempt to process a new method to obtain pluripotency from goat somatic cells in vitro under fully chemically defined condition. The results showed that chemically induced pluripotent stem cells-like cells (CiPSC-like cells) colonies were generated from goat ear fibroblasts by fully small-molecule compounds. Those three dimensions colonies were similar with mouse iPSCs in morphology and had strong positive alkaline phosphatase (AP) activity and expressed pluripotency related genes OCT4, SOX2, NANOG, CDH1, TDGF, GDF3, DAX1, REX1, which determined by RT-PCR. Those colonies could also differentiate into different cell types derived from three germ layers proved by RT-PCR and immunofluorescence assays. The expression of glycolysis-related genes about PGAM1, KPYM2 and HXK2 in CiPSC-like colonies formation groups was significantly higher than their parental fibroblasts, but not in the non-CiPSC-like colonies formation group. The expression of histone acetylation and methylation-related genes, HAT1 and SMYD3, was not significantly up-regulated within different groups compared to their parental fibroblasts, respectively. Yet, the expression of histone methylation-related gene, KDM5B, was significantly up-regulated on the cells from non-colonies formation group compared to parental fibroblasts, but the expression of KDM5B of the cells from CiPSC-like cell colonies was not significantly difference compared to that of parental fibroblasts. In conclusion, this is the first report that CiPSC-like cells could be generated in vitro from goat rather than just mouse under fully chemically defined condition. The generation of CiPSC-like colonies may be depended on the correct modification of energy metabolism and histone epigenetic during the reprogramming, rather than just the over-expression of those pluripotency-related genes. This study will strongly support us to further establish the stable goat CiPSC lines without any integration of exogenous genes.


Assuntos
Histonas/metabolismo , Células-Tronco Pluripotentes Induzidas/citologia , Mitocôndrias/metabolismo , Fatores de Transcrição/genética , Animais , Diferenciação Celular/genética , Células Cultivadas , Reprogramação Celular , Proteínas de Ligação a DNA/genética , Metabolismo Energético , Fibroblastos/citologia , Cabras/metabolismo , Histona Desmetilases com o Domínio Jumonji/genética , Camundongos , Regulação para Cima
18.
Pancreatology ; 18(7): 822-832, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30201439

RESUMO

BACKGROUND: Cantharidin is an inhibitor of protein phosphatase 2 A (PP2A), and has been frequently used in clinical practice. In our previous study, we proved that cantharidin could arrest cell cycle in G2/M phase. Since cells at G2/M phase are sensitive to radiotherapy, in the present study, we investigated the radiotherapy-sesitization effect of cantharidin and the potential mechanisms involved. METHODS: Cell growth was determined by MTT assay. Cell cycle was evaluated by flow cytometry. DNA damage was visualized by phospho-Histone H2A.X staining. Expression of mRNA was tested by microarray assay and real-time PCR. Clinical information and RNA-Seq expression data were derived from The Cancer Genome Atlas (TCGA) pancreatic cancer cohort. Survival analysis was obtained by Kaplan-Meier estimates. RESULTS: Cantharidin strengthened the growth inhibition effect of irradiation. Cantharidin drove pancreatic cancer cells out of quiescent G0/G1 phase and arrested cell cycle in G2/M phase. As a result, cantharidin strengthened DNA damage which was induced by irradiation. Moreover, cantharidin repressed expressions of several genes participating in DNA damage repair, including UBE2T, RPA1, GTF2HH5, LIG1, POLD3, RMI2, XRCC1, PRKDC, FANC1, FAAP100, RAD50, RAD51D, RAD51B and DMC1, through JNK, ERK, PKC, p38 and/or NF-κB pathway dependent manners. Among these genes, worse overall survival for pancreatic cancer patients were associated with high mRNA expressions of POLD3, RMI2, PRKDC, FANC1, RAD50 and RAD51B, all of which could be down-regulated by cantharidin. CONCLUSION: Cantharidin can sensitize pancreatic cancer cells to radiotherapy. Multiple mechanisms, including cell cycle regulation, enhanced DNA damage, and inhibited DNA damage repair, may be involved.


Assuntos
Cantaridina/farmacologia , Radiossensibilizantes/farmacologia , Radioterapia , Ciclo Celular , Divisão Celular/efeitos dos fármacos , Divisão Celular/efeitos da radiação , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos da radiação , Dano ao DNA , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Neoplasias Pancreáticas , Proteína Fosfatase 2/antagonistas & inibidores
19.
Methods Mol Biol ; 1830: 81-92, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30043365

RESUMO

Transcription factors are pivotal for the control of development and the response of organisms to changes in the environment. Therefore, a detailed understanding of their functions is of central importance for biology. Over the years, different experimental methods have been developed to study the activities of transcription factors in plants. These methods include perturbation assays, where the activity of a given transcription factor is disrupted and subsequently, the resulting effects are monitored using molecular, genomic, or physiological approaches. Perturbation assays can also be used to distinguish primary roles of transcription factors of interest from secondary effects. Thus, molecular genetic experiments after perturbation can be advantageous or even necessary for the precise understanding of transcription factor function at a certain stage of plant development or in a single tissue or organ type. In this chapter, we describe several commonly used techniques to knock down transcription factor activities and provide detailed information on how those techniques are employed in the model plant Arabidopsis thaliana.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Técnicas de Silenciamento de Genes/métodos , Fatores de Transcrição/metabolismo , Arabidopsis/efeitos dos fármacos , Dexametasona/farmacologia , Estradiol/farmacologia , Etanol/farmacologia , Especificidade de Órgãos/efeitos dos fármacos , Regiões Promotoras Genéticas , Interferência de RNA , Transcrição Gênica/efeitos dos fármacos
20.
Reprod Domest Anim ; 53(5): 1191-1199, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-29974990

RESUMO

To further promote the early development of porcine embryos and capture "naïve" pluripotent state within blastocyst, the experiment explored the effects of lysophosphatidic acid (LPA) on the early development of porcine parthenogenetic embryos and the expression of pluripotency relevant genes. The results showed that the addition of 50 µM LPA significantly improved parthenogenetic embryo cleavage rate (82.7% vs. 74.7%, p < 0.05), blastocyst rate (24.5% vs. 11.3%, p < 0.05) and blastocyst cell count (56 ± 7.9 vs. 42 ± 1.0, p < 0.05) than that of the control group. In addition, immunostaining experiment determined that the fluorescence intensity of OCT4 was also significantly higher than that of the control group. The quantitative real-time polymerase chain reaction (qRT-PCR) test revealed that addition of 50 µM LPA could significantly enhance the expression level of pluripotent gene OCT4 and trophoblast marker genes CDX2, however, decrease the expression of primitive hypoblast marker gene GATA4. The results also indicated that LPA might decrease the expression of GATA4 through the ROCK signalling pathway. For further investigating the effect of the addition of LPA on the expression of "primed" and "naïve" genes, we also detected the expression of those pluripotency-related genes by qRT-PCR. The results showed addition of LPA had no significant effect on the expression of "naïve" pluripotent genes, but it was able to significantly decrease the expression of "primed" pluripotent genes, NODAL and Activin-A; furthermore, it also could significantly improve the expression of OCT4 and c-Myc which act as two important ES cell renewal factors. Above all, the addition of LPA can facilitate the early development of porcine parthenogenetic embryos, which may be able to benefit for capturing "naïve" pluripotency in vitro through inhibiting "primed" pluripotency.


Assuntos
Blastocisto/efeitos dos fármacos , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Marcadores Genéticos , Lisofosfolipídeos/farmacologia , Partenogênese , Animais , Células-Tronco Embrionárias/citologia , Transdução de Sinais/efeitos dos fármacos , Suínos
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