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The MADS-box gene plays an important role in plant growth and development. As an important vegetable of Cucurbitaceae, chayote has great edible and medicinal value. So far, there is little molecular research on chayote, and there are no reports on the MADS-box transcription factor of chayote. In this study, the MADS-box gene family of chayote was analyzed for the first time, and a total of 70 MADS-box genes were identified, including 14 type I and 56 type II MICK MADS genes. They were randomly distributed on 13 chromosomes except for chromosome 11. The light response element, hormone response element and abiotic stress response element were found in the promoter region of 70 MADS genes, indicating that the MADS gene can regulate the growth and development of chayote, resist abiotic stress, and participate in hormone response; GO and KEGG enrichment analysis also found that SeMADS genes were mainly enriched in biological regulation and signal regulation, which further proved the important role of MADS-box gene in plant growth and development. The results of collinearity showed that segmental duplication was the main driving force of MADS gene expansion in chayote. RNA-seq showed that the expression levels of SeMADS06, SeMADS13, SeMADS26, SeMADS28, SeMADS36 and SeMADS37 gradually increased with the growth of chayote, indicating that these genes may be related to the development of root tubers of 'Tuershao'. The gene expression patterns showed that 12 SeMADS genes were specifically expressed in the male flower in 'Tuershao' and chayote. In addition, SeMADS03 and SeMADS52 may be involved in regulating the maturation of male flowers of 'Tuershao' and chayote. SeMADS21 may be the crucial gene in the development stage of the female flower of 'Tuershao'. This study laid a theoretical foundation for the further study of the function of the MADS gene in chayote in the future.
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Cucurbitaceae , Proteínas de Domínio MADS , Proteínas de Domínio MADS/genética , Proteínas de Domínio MADS/metabolismo , Genoma de Planta , Flores/metabolismo , Fatores de Transcrição/metabolismo , Cucurbitaceae/genética , Filogenia , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismoRESUMO
MOTIVATION: Microscopy technology plays important roles in many biological research fields. Solvent-cleared brain high-resolution (HR) 3D image reconstruction is an important microscopy application. However, 3D microscopy image generation is time-consuming and expensive. Therefore, we have developed a deep learning framework (DeepS) for both image optical sectioning and super resolution microscopy. RESULTS: Using DeepS to perform super resolution solvent-cleared mouse brain microscopy 3D image yields improved performance in comparison with the standard image processing workflow. We have also developed a web server to allow online usage of DeepS. Users can train their own models with only one pair of training images using the transfer learning function of the web server. AVAILABILITYAND IMPLEMENTATION: http://deeps.cibr.ac.cn. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.
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Aprendizado Profundo , Microscopia , Animais , Camundongos , Computadores , Processamento de Imagem Assistida por Computador , Imageamento TridimensionalRESUMO
Chayote (Sechium edule) produces edible tubers with high starch content after 1 year of growth but the mechanism of chayote tuberization remains unknown. 'Tuershao', a chayote cultivar lacking edible fruits but showing higher tuber yield than traditional chayote cultivars, was used to study tuber formation through integrative analysis of the metabolome and transcriptome profiles at three tuber-growth stages. Starch biosynthesis- and galactose metabolism-related genes and metabolites were significantly upregulated during tuber bulking, whereas genes encoding sugars will eventually be exported transporter (SWEET) and sugar transporter (SUT) were highly expressed during tuber formation. Auxin precursor (indole-3-acetamide) and ethylene precursor, 1-aminocyclopropane-1-carboxylic acid, were upregulated, suggesting that both hormones play pivotal roles in tuber development and maturation. Our data revealed a similar tuber-formation signaling pathway in chayote as in potatoes, including complexes BEL1/KNOX and SP6A/14-3-3/FDL. Down-regulation of the BEL1/KNOX complex and upregulation of 14-3-3 protein implied that these two complexes might have distinct functions in tuber formation. Finally, gene expression and microscopic analysis indicated active cell division during the initial stages of tuber formation. Altogether, the integration of transcriptome and metabolome analyses unraveled an overall molecular network of chayote tuberization that might facilitate its utilization.
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Cucurbitaceae , Solanum tuberosum , Cucurbitaceae/genética , Regulação da Expressão Gênica de Plantas , Metaboloma , Tubérculos/metabolismo , Solanum tuberosum/metabolismo , Amido/metabolismo , TranscriptomaRESUMO
Altered expression of the E3 ubiquitin ligase UBE3A, which is involved in protein degradation through the proteasome-mediated pathway, is associated with neurodevelopmental and behavioral defects observed in Angelman syndrome (AS) and autism. However, little is known about the neuronal function of UBE3A and the pathogenesis of UBE3A-associated disorders. To understand the in vivo function of UBE3A in the nervous system, we generated multiple mutations of ube3a, the Drosophila ortholog of UBE3A. We found a significantly increased number of total boutons and satellite boutons in conjunction with compromised endocytosis in the neuromuscular junctions (NMJs) of ube3a mutants compared to the wild type. Genetic and biochemical analysis showed upregulation of bone morphogenetic protein (BMP) signaling in the nervous system of ube3a mutants. An immunochemical study revealed a specific increase in the protein level of Thickveins (Tkv), a type I BMP receptor, but not other BMP receptors Wishful thinking (Wit) and Saxophone (Sax), in ube3a mutants. Ube3a was associated with and specifically ubiquitinated lysine 227 within the cytoplasmic tail of Tkv, and promoted its proteasomal degradation in Schneider 2 cells. Negative regulation of Tkv by Ube3a was conserved in mammalian cells. These results reveal a critical role for Ube3a in regulating NMJ synapse development by repressing BMP signaling. This study sheds new light onto the neuronal functions of UBE3A and provides novel perspectives for understanding the pathogenesis of UBE3A-associated disorders.
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Síndrome de Angelman/genética , Proteínas de Drosophila/biossíntese , Proteínas de Drosophila/genética , Neurônios/metabolismo , Proteínas Serina-Treonina Quinases/biossíntese , Receptores de Superfície Celular/biossíntese , Ubiquitina-Proteína Ligases/genética , Síndrome de Angelman/patologia , Animais , Proteínas Morfogenéticas Ósseas/biossíntese , Proteínas Morfogenéticas Ósseas/genética , Modelos Animais de Doenças , Drosophila/genética , Endocitose/genética , Regulação da Expressão Gênica/genética , Humanos , Junção Neuromuscular/genética , Junção Neuromuscular/patologia , Neurônios/patologia , Proteínas Serina-Treonina Quinases/genética , Receptores de Superfície Celular/genética , Transdução de Sinais , Sinapses/genética , Ubiquitina-Proteína Ligases/biossínteseRESUMO
Drought significantly affects crop productivity and poses a considerable threat to agricultural ecosystems. Plant growth-promoting bacteria (PGPB) and plant microbiome play important roles in improving drought resistance and plant performance. However, the response of the rhizosphere microbiota to PGPB during the development of plants and the interaction between inoculum, microbiota, and plants under drought stress remain to be explored. In the present study, we used culturomic, microbiomic, and metabonomic analyses to uncover the mechanisms by which Sphingomonas sp. Hbc-6, a PGPB, promotes Arabidopsis growth and enhances drought resistance. We found that the rhizosphere microbiome assembly was interactively influenced by developmental stage, Hbc-6, and drought; the bacterial composition exhibited three patterns of shifts with developmental stage: resilience, increase, and decrease. Drought diminished microbial diversity and richness, whereas Hbc-6 increased microbial diversity and helped plants recruit specific beneficial bacterial taxa at each developmental stage, particularly during the bolting stage. Some microorganisms enriched by Hbc-6 had the potential to promote carbon and nitrogen cycling processes, and 86.79â¯% of the isolated strains exhibited PGP characteristics (for example Pseudomonas sp. TA9). They jointly regulated plant physiological metabolism (i.e., upregulated drought resistant-facilitating substances and reduced harmful substances), thereby stimulating the growth of Arabidopsis and increasing plant biomass under drought stress conditions. Collectively, these results indicate that Hbc-6 mediates plant growth and drought resistance by affecting the microbiome. The study thus provides novel insights and strain resources for drought-resistant, high-yielding crop cultivation and breeding.
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Objective: This study aimed to identify clinical characteristics associated with the prevalence of progressive pulmonary fibrosis (PPF) in interstitial lung disease (ILD) and to develop a prognostic nomogram model for clinical use. Methods: In this single-centered, retrospective study, we enrolled ILD patients with relatively comprehensive clinical data and assessed the incidence of PPF within a year using collected demographics, laboratory data, high-resolution computed tomography (HRCT), and pulmonary function test (PFT) results. We used a training cohort of ILD patients to identify early predictors of PPF and then validated them in an internal validation cohort and subsets of ILD patients using a multivariable logistic regression analysis. A prognostic nomogram was formulated based on these predictors, and the accuracy and efficiency were evaluated using the area under the receiver operating characteristic curve (AUC), calibration plot, and decision curve analysis (DCA). Results: Among the enrolled patients, 120 (39.09%) cases had connective tissue disease-associated interstitial lung disease (CTD-ILD), 115 (37.46%) had non-idiopathic pulmonary fibrosis idiopathic interstitial pneumonia (non-IPF IIP), and 35 (11.4%) had hypersensitivity pneumonitis (HP). Overall, 118 (38.4%) cases experienced pulmonary fibrosis progression. We found that baseline DLco% pred (OR 0.92; 95% CI, 8.93-0.95) was a protective factor for ILD progression, whereas combined pneumonia (OR 4.57; 95% CI, 1.24-18.43), modified Medical Research Council dyspnea score (mMRC) (OR 4.9; 95% CI, 2.8-9.5), and high-resolution computed tomography (HRCT) score (OR 1.22; 95% CI, 1.07-1.42) were independent risk factors for PPF. The AUC of the proposed nomogram in the development cohort was 0.96 (95% CI, 0.94, 0.98), and the calibration plot showed good agreement between the predicted and observed incidence of PPF (Hosmer-Lemeshow test: P = 0.86). Conclusion: ILD patients with combined pneumonia, low baseline DLco% pred, high mMRC marks, and high HRCT scores were at higher risk of progression. This nomogram demonstrated good discrimination and calibration, indicating its potential utility for clinical practice.
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Purpose: Stage T1 esophageal cancer (EC) with distant metastasis (DM) is rare and poorly understood. In this study, we aimed to construct and validate a novel nomogram for predicting the probability of DM in T1 EC patients. Methods: A total of 1,663 eligible T1 EC patients were enrolled from the Surveillance, Epidemiology, and End Results (SEER) database between 2004 and 2015. The patients were randomly divided into training and validation cohorts. Univariate and multivariate logistic analyses in the training cohort were used to identify risk factors related to DM, and then these risk factors were applied to construct the nomogram. Receiver operating characteristic (ROC) curves, the area under the curve (AUC), calibration plots, the Hosmer-Lemeshow (HL) test, and decision curve analysis (DCA) were used to evaluate the nomogram. Results: Among the 1,663 patients identified, 143 (8.6%) had DM. Five risk factors (tumor location, lymph node status, tumor length, T1 subtype, and grade) were significant predictors of DM. The AUC values were 0.828 and 0.851 in the training cohort and validation cohort, respectively, revealing good discrimination. The calibration plots in the training cohort and validation cohort both showed good consistency. DCA showed that the nomogram was clinically effective. In addition, the nomogram has a good risk stratification ability to identify patients with different risks according to the nomogram score. In terms of survival analysis, univariate and multivariate Cox analyses showed that age, race, tumor length, grade, lymph node status, M stage and treatment were significant prognostic factors for overall survival (OS). For cancer-specific survival (CSS), the independent prognostic factors were age, tumor length, histology, grade, lymph node status, M stage and treatment. Conclusion: The nomogram could effectively predict the probability of DM in T1 EC patients. It can aid clinicians in detecting high-risk patients and making individual clinical decisions.
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The mechanism by which closely related species can coexist is a central factor in the stability of ecological communities. The larch adelgid (Adelges laricis laricis) and the eastern spruce adelgid (Adelges (Sacchiphantes) abietis) have both been found on the branches of Picea koraiensis in China. These two adelgids exhibit strong infectivity and readily induce the formation of 'fish scale-like' and 'pineapple-like' galls with branch parasitism rates of between 75.01 ± 7.03 and 88.02 ± 4.39%. Interestingly, the gall tissues in which these two gall-inducing insects were found to be coexisting were discovered at a rate of ~0.2% in the studied populations. The weight and number of gall chambers as well as the number of adelgids in the 'fish scale-like' side were higher than those in the 'pineapple-like' side. Furthermore, compared with the normal branches, a diterpene neoabietic acid was found at elevated concentrations in the gall tissues, with especially high concentrations seen in the tissues of the co-occupied galls. Neoabietic acid exhibited strong antibacterial activities against Bacillus spp. isolated from the branches of P. koraiensis, as well as potent antifungal activity against the hyphal growth of Fusarium graminearum JMY-1, which was obtained from the gall tissues. Our result provides evidence that the coexistence of the two closely related species could be explained by alterations of the host tissues by the insects resulting in increased concentrations of the antimicrobial agent.
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Anti-Infecciosos , Diterpenos , Picea , Animais , Antibacterianos , Anti-Infecciosos/farmacologia , Diterpenos/farmacologia , InsetosRESUMO
Drought poses a serious threat to plant growth. Plant growth-promoting bacteria (PGPB) have great potential to improve plant nutrition, yield, and drought tolerance. Sphingomonas is an important microbiota genus that is extensively distributed in the plant or rhizosphere. However, the knowledge of its plant growth-promoting function in dry regions is extremely limited. In this study, we investigated the effects of PGPB Sphingomonas sp. Hbc-6 on maize under normal conditions and drought stress. We found that Hbc-6 increased the biomass of maize under normal conditions and drought stress. For instance, the root fresh weight and shoot dry weight of inoculated maize increased by 39.1% and 34.8% respectively compared with non-inoculated plant, while they increased by 61.3% and 96.3% respectively under drought conditions. Hbc-6 also promoted seed germination, maintained stomatal morphology and increased chlorophyll content so as to enhance photosynthesis of plants. Hbc-6 increased antioxidant enzyme (catalase, superoxide, peroxidase) activities and osmoregulation substances (proline, soluble sugar) and up-regulated the level of beneficial metabolites (resveratrol, etc.). Moreover, Hbc-6 reshaped the maize rhizosphere bacterial community, increased its richness and diversity, and made the rhizosphere bacterial community more complex to resist stress; Hbc-6 could also recruit more potentially rhizosphere beneficial bacteria which might promote plant growth together with Hbc-6 both under normal and drought stress. In short, Hbc-6 increased maize biomass and drought tolerance through the above ways. Our findings lay a foundation for exploring the complex mechanisms of interactions between Sphingomonas and plants, and it is important that Sphingomonas sp. Hbc-6 can be used as a potential biofertilizer in agricultural production, which will assist finding new solutions for improving the growth and yield of crops in arid areas.
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Latex is a type of sticky endogenous fluids derived from diverse plants including Euphorbia fischeriana, and is of great scientific and commercial values. In the current study, it was demonstrated that the latex extracted from E. fischeriana strongly respelled the growth of cotton bollworm, Helicoverpa armigera. Using spectroscopic methods, HPLC, and GC-MS analyses, six aliphatic tigliane diterpenoids were isolated from the latex of E. fischeriana, among which three compounds (2, 3, and 5) were new. Two major compounds (1 and 4) exhibited remarkable antifeedant activity against H. armigera, with EC50 values at 2.59 and 15.32 µg/cm2, respectively. In addition, the quantification analysis of diterpenoids in different organs indicated that 4 was the most abundant constituent and was highly accumulated in the latex. Collectively, the current study highlighted that the diterpenoids in latex of E. fischeriana had a considerable antifeedant function against H. armigera, which might be employed for the future development of natural insecticides for organic farming.
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Diterpenos/química , Diterpenos/farmacologia , Euphorbia/química , Látex/química , Animais , Cromatografia Líquida de Alta Pressão , Diterpenos/análise , Euphorbia/metabolismo , Cromatografia Gasosa-Espectrometria de Massas , Inseticidas/química , Inseticidas/farmacologia , Estrutura Molecular , Mariposas/efeitos dos fármacos , Forbóis/químicaRESUMO
Conspicuous color is a common trait of foliar galls, but their relationship with gall-inducing insects is unknown. Red and green galls were taken from sunny or shady parts of peach species Prunus persica (L.) Batsch. f. rubro-plena Schneid with peach aphid Tuberocephalus momonis (Matsumura) infestation. We found that the loss of photosynthetic pigments was associated with the conspicuous coloration of green gall tissues. The concentrations of anthocyanins significantly increased following ultraviolet (UV) irradiation of green gall tissues, suggesting that accumulation of anthocyanins in red galls is related to ultraviolet B and C (UVB and UVC) radiation. The expression of structural genes related to the biosynthesis of chlorogenic acid and malic acid benzoate was increased in all gall tissues and negatively correlated with the expression profiles of certain genes associated with photosynthetic biosynthesis, indicating that the increased transcript levels of the phenylpropanoid pathway might cause loss of photosynthetic efficiency in the gall tissues. Transcriptome and quantitative reverse transcription PCR analyses revealed that MYB transcription factors that up-regulate the biosynthesis of anthocyanins in red gall tissues might be activated by both UVB and UVC exposure. Comet assays suggest that green and red gall tissues have similar DNA damage following UV irradiation. No obvious effect of the up-regulated compounds on the growth of the peach aphid was observed. Interestingly, peach aphids under leaves painted with anthocyanins had lower mortality following UV irradiation than those in controls. These results suggest that the anthocyanins in red gall tissues have a defensive function for the peach aphid, protecting it against UV radiation.
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Afídeos , Prunus persica , Animais , Antocianinas , Prunus persica/genética , Raios Ultravioleta , Regulação para CimaRESUMO
The larch adelgid Adelges laricis laricis Vallot is a specialist insect parasite of Picea koraiensis (Korean spruce) and forms fish scale-like galls that damage the growth of the host plants. Our investigation reveals that both these galls and the fruits (cones) of P. koraiensis display lower concentrations of phytosynthetic pigments and accumulate anthocyanin cyanidin-3-O-glucoside and soluble sugars in the mature stages. Interestingly, high concentrations of 6-benzylaminopurine (BAP) both in the cauline gall tissues and in the larch adelgids themselves (4064.61 ± 167.83 and 3655.42 ± 210.29 ng/g FW, respectively), suggested that this vital phytohormone may be synthesized by the insects to control the development of gall tissues. These results indicate that the galls and cones are sink organs, and the development of gall tissues is possibly regulated by phytohormones in a way similar to that of the growth of cones. The concentrations of phytohormones related to growth [indole-3-acetic acid (IAA), cytokinins (CTK), and gibberellins (GAs)] and defense [salicylic acid (SA)], as well as SA-related phenolics [benzoic acid (BA) and p-hydroxybenzoic acid (pHBA)] in gall tissues were positively correlated with those in cones during the development stage. The levels of 1-aminocyclopropane-1-carboxylic acid (ACC) in the developmental stage of the cones correlates negatively with their concentrations in the gall tissues (R = -0.92, p < 0.001), suggesting that downregulation of ACC might be the reason why galls are not abscised after a year. Our results provide a new perspective on the potential mechanism of the development of cauline galls on P. koraiensis, which are regulated by phytohormones.
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Mutations in long-chain acyl-CoA synthetase 4 (ACSL4) are associated with non-syndromic X-linked intellectual disability (ID). However, the neural functions of ACSL4 and how loss of ACSL4 leads to ID remain largely unexplored. We report here that mutations in Acsl, the Drosophila ortholog of human ACSL3 and ACSL4, result in developmental defects of the mushroom body (MB), the center of olfactory learning and memory. Specifically, Acsl mutants show fewer MB neuroblasts (Nbs) due to reduced proliferation activity and premature differentiation. Consistently, these surviving Nbs show reduced expression of cyclin E, a key regulator of the G1- to S-phase cell cycle transition, and nuclear mislocalization of the transcriptional factor Prospero, which is known to repress self-renewal genes and activate differentiating genes. Furthermore, RNA-seq analysis reveals downregulated Nb- and cell-cycle-related genes and upregulated neuronal differentiation genes in Acsl mutant Nbs. As Drosophila Acsl and human ACSL4 are functionally conserved, our findings provide novel insights into a critical and previously unappreciated role of Acsl in neurogenesis and the pathogenesis of ACSL4-related ID.
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Coenzima A Ligases/genética , Drosophila melanogaster/genética , Deficiência Intelectual/genética , Mutação , Neurônios/citologia , Homologia de Sequência do Ácido Nucleico , Transporte Ativo do Núcleo Celular , Animais , Diferenciação Celular , Núcleo Celular/metabolismo , Proliferação de Células/genética , Ciclina E/metabolismo , Regulação da Expressão Gênica , Humanos , Transcrição GênicaRESUMO
OBJECTIVE: Time window is a key factor in the treatment of lung injury by mesenchymal stem cells (MSC) transplantation. This study was aimed to analyze the engraftment and differentiation behavior of MSC transplanted at different time points after lung irradiation, and the possible mechanisms were discussed. MATERIALS AND METHODS: The thorax of C57BL/6 mice was exposed to 1400 cGy, then Flk-1(+)MSCs from enhanced green fluorescent protein C57BL/6 mice were systemically injected into C57BL/6 mice at 4 hours, 60 days, and 120 days post thoracic exposure, respectively. The engraftment and differentiation of Flk-1(+)MSC transplanted at different time points were evaluated. Lung tissue was collected and analyzed for fibrosis. Expression of transforming growth factor (TGF)-beta1 in the lung was qualified by semi-quantitative real-time reverse transcription polymerase chain reaction. In vitro, Flk-1(+)MSCs were cultured in epithelium induction media, together with damaged primary lung cells, supernatants of radiation-injured lung cells, or TGF-beta1 to find the possible factors that might effect Flk-1(+)MSC differentiation. RESULTS: Cells injected immediately after injury were shown to differentiate into functional lung cells, such as epithelial and endothelial cells. Cells injected 2 months later were mostly located in the interstitial area and appeared as myofibrocyte. The in vivo lung microenvironments at different time points after injury were different from each other, especially TGF-beta1 expression. We demonstrated that cytokines secreted by irradiated lung cells could inhibit differentiation of Flk-1(+)MSCs into epithelial cells in vitro. CONCLUSIONS: Flk-1(+)MSCs injected into the lung immediately after irradiation could differentiate into functional lung cells, while those injected at later stage after irradiation would be involved in fibrosis development. Thus our in vivo and in vitro studies demonstrated that differentiation of Flk-1(+)MSCs is controlled by the microenvironment.
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Lesão Pulmonar , Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais , Lesões Experimentais por Radiação/terapia , Pneumonite por Radiação/terapia , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/biossíntese , Animais , Diferenciação Celular , Células Epiteliais/efeitos dos fármacos , Imuno-Histoquímica , Pulmão/patologia , Pulmão/efeitos da radiação , Células-Tronco Mesenquimais/citologia , Camundongos , Camundongos Endogâmicos C57BL , RNA Mensageiro/biossíntese , Fatores de Tempo , Fator de Crescimento Transformador beta/metabolismo , Fator de Crescimento Transformador beta/farmacologia , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/genéticaRESUMO
Flk-1(+)CD31(-)CD34(-) mesenchymal stem cells (MSCs) are multipotent cells with extensive proliferation ability and immunomodulatory function. On the basis of our previous work showing their potential application in tissue engineering and immune diseases, we designed a preclinical and Phase I trial to evaluate the feasibility and safety of intravenous infusion of these cells after ex vivo expansion. Rhesus monkey and human Flk-1(+)CD31(-)CD34(-) MSCs were isolated from bone marrow and passaged a maximum of six passages. Karyotype analysis showed Flk-1(+)CD31(-)CD34(-) MSCs remained normally diploid within six passages of expansion. Expanded cells were transplanted into rhesus monkeys and human volunteers, respectively. During the infusion of these cells, the life signs of the recipients were normal. Laboratory tests for blood, bone marrow, kidney, and liver function were conducted and no significant changes were observed before and after cell infusion. Identification of the sry sequence from the male donor in female recipients showed that allogeneic rhesus Flk- 1(+)CD31(-)CD34(-) MSCs were capable of homing to and establishing residence within the recipients' bone marrow. No significant changes were observed in lymphocyte cell subpopulation before and after infusion. These result showed that Flk-1(+)CD31(-)CD34(-) MSCs have no obvious side effects after intravenous administration, encouraging investigators to perform further studies in stem cell therapy.
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Antígenos CD34/metabolismo , Células da Medula Óssea/citologia , Haplorrinos/metabolismo , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/metabolismo , Molécula-1 de Adesão Celular Endotelial a Plaquetas/metabolismo , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/metabolismo , Adulto , Animais , Antígenos de Superfície/metabolismo , Proliferação de Células , Células Cultivadas , Quimerismo , Cromossomos de Mamíferos/genética , Estudos de Viabilidade , Feminino , Humanos , Infusões Intravenosas , Cariotipagem , Testes de Função Renal , Testes de Função Hepática , Masculino , Transplante de Células-Tronco Mesenquimais , Transplante AutólogoRESUMO
Our previous studies have revealed that amyloid ß (Aß)-binding alcohol dehydrogenase (ABAD) decoy peptide antagonizes Aß42-induced neurotoxicity. However, whether it improves oxidative stress injury remains unclear. In this study, a recombinant adenovirus constitutively secreting and expressing Aß-ABAD decoy peptide (rAAV/ABAD-DP-6His) was successfully constructed. Our results showed that rAAV/ABAD-DP-6His increased superoxide dismutase activity in hydrogen peroxide-induced oxidative stress-mediated injury of PC12 cells. Moreover, rAAV/ABAD-DP-6His decreased malondialdehyde content, intracellular Ca(2+) concentration, and the level of reactive oxygen species. rAAV/ABAD-DP-6His maintained the stability of the mitochondrial membrane potential. In addition, the ATP level remained constant, and apoptosis was reduced. Overall, the results indicate that rAAV/ABAD-DP-6His generates the fusion peptide, Aß-ABAD decoy peptide, which effectively protects PC12 cells from oxidative stress injury induced by hydrogen peroxide, thus exerting neuroprotective effects.
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Amyloid ß peptide binding alcohol dehydrogenase (ABAD) decoy peptide (DP) can competitively antagonize binding of amyloid ß peptide to ABAD and inhibit the cytotoxic effects of amyloid ß peptide. Based on peptide aptamers, the present study inserted ABAD-DP into the disulfide bond of human thioredoxin (TRX) using molecular cloning technique to construct a fusion gene that can express the TRX1-ABAD-DP-TRX2 aptamer. Moreover, adeno-associated virus was used to allow its stable expression. Immunofluorescent staining revealed the co-expression of the transduced fusion gene TRX1-ABAD-DP-TRX2 and amyloid ß peptide in NIH-3T3 cells, indicating that the TRX1-ABAD-DP-TRX2 aptamer can bind amyloid ß peptide within cells. In addition, cell morphology and MTT results suggested that TRX1-ABAD-DP-TRX2 attenuated amyloid ß peptide-induced SH-SY5Y cell injury and improved cell viability. These findings confirmed the possibility of constructing TRX-based peptide aptamer using ABAD-DP. Moreover, TRX1-ABAD-DP-TRX2 inhibited the cytotoxic effect of amyloid ß peptide.
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Low oxygen tension is a potent differentiation inducer of numerous cell types and an effective stimulus of many gene expressions. Here, we described that under 8% O(2), bone marrow stromal cells (MSCs) exhibited proliferative and morphologic changes. The level of differentiated antigen H-2Dd and the number of G(2)/S/M phase cells increased evidently under 8% O(2) condition. Also, the proportion of wide, flattened, and epithelial-like cells (which were alkaline phosphatase staining positive) in MSCs increased significantly. When cultured in adipogenic medium, there was a 5- to 6-fold increase in the number of lipid droplets under hypoxic conditions compared with that in normoxic culture. We also demonstrated the existence of MSC differentiation under hypoxic conditions by electron microscopy. Expression of Oct4 was inhibited under 8% O(2) condition, but after adipocyte differentiation in normoxic culture and hypoxia-mimicking agents cobalt chloride (CoCl(2)) and deferoxamine mesylate (DFX) treatments, Oct4 was still expressed in MSCs. These results indicate hypoxia accelerates MSC differentiation and hypoxia and hypoxia-mimicking agents exert different effects on MSC differentiation.