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1.
Theranostics ; 11(16): 7879-7895, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34335970

RESUMO

Rationale: Previous studies have shown that human embryonic stem cell-derived cardiomyocytes improved myocardial recovery when administered to infarcted pig and non-human primate hearts. However, the engraftment of intramyocardially delivered cells is poor and the effectiveness of clinically relevant doses of human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) in large animal models of myocardial injury remains unknown. Here, we determined whether thymosin ß4 (Tb4) could improve the engraftment and reparative potency of transplanted hiPSC-CMs in a porcine model of myocardial infarction (MI). Methods: Tb4 was delivered from injected gelatin microspheres, which extended the duration of Tb4 administration for up to two weeks in vitro. After MI induction, pigs were randomly distributed into 4 treatment groups: the MI Group was injected with basal medium; the Tb4 Group received gelatin microspheres carrying Tb4; the CM Group was treated with 1.2 × 108 hiPSC-CMs; and the Tb4+CM Group received both the Tb4 microspheres and hiPSC-CMs. Myocardial recovery was assessed by cardiac magnetic resonance imaging (MRI), arrhythmogenesis was monitored with implanted loop recorders, and tumorigenesis was evaluated via whole-body MRI. Results: In vitro, 600 ng/mL of Tb4 protected cultured hiPSC-CMs from hypoxic damage by upregulating AKT activity and BcL-XL and promoted hiPSC-CM and hiPSC-EC proliferation. In infarcted pig hearts, hiPSC-CM transplantation alone had a minimal effect on myocardial recovery, but co-treatment with Tb4 significantly enhanced hiPSC-CM engraftment, induced vasculogenesis and the proliferation of cardiomyocytes and endothelial cells, improved left ventricular systolic function, and reduced infarct size. hiPSC-CM implantation did not increase incidence of ventricular arrhythmia and did not induce tumorigenesis in the immunosuppressed pigs. Conclusions: Co-treatment with Tb4-microspheres and hiPSC-CMs was safe and enhanced the reparative potency of hiPSC-CMs for myocardial repair in a large-animal model of MI.


Assuntos
Infarto do Miocárdio/terapia , Miócitos Cardíacos/metabolismo , Timosina/farmacologia , Animais , Diferenciação Celular , Proliferação de Células , Células Cultivadas , China , Modelos Animais de Doenças , Células Endoteliais/patologia , Humanos , Células-Tronco Pluripotentes Induzidas/metabolismo , Infarto do Miocárdio/metabolismo , Miocárdio/patologia , Regeneração , Transplante de Células-Tronco/métodos , Suínos , Timosina/metabolismo , Timosina/fisiologia
2.
Artigo em Inglês | MEDLINE | ID: mdl-34279752

RESUMO

OBJECTIVES: We sought to distinguish area at risk from salvage myocardial zone and to predict left ventricle functional recovery in the convalescent stage by Texture Analysis (TA) of T2-Mapping. METHODS: One hundred and six patients diagnosed with AMI and treated with percutaneous coronary intervention (PCI) underwent acute cardiac magnetic resonance imaging (CMR) and 45 of whom had a subsequent CMR scan following recovery. Cine imaging, T2-Mapping, T2-weighted STIR imaging, and LGE imaging were performed. In the texture analysis, regions of interest (infarcted, salvageable, and remote) were drawn by two blinded, independent readers. RESULTS: Seven independent texture features on T2-Mapping were selected: Perc.50%, S(2,2)InvDfMom, S(2.-2)AngScMom, S(4,0)Entropy, 45dgrLngREmph, 45dgr_Fraction and 135dr_GLevNonU. Among them, the average value of 135dr_GLevNonU in the infarct zone, AAR zone, and the remote zone was: 61.96±26.03, 31.811±18.933 and 99.839±26.231, respectively. Additionally, 135dr_GLevNonU provided the highest area under the curve (AUC) from the receiver operating characteristic curve (ROC curve) for distinguishing AAR from the infarct zone in each subgroup (all patients, patients with MVO and)were 0.845 ± 0.052 0.855 ± 0.083 and 0.845 ± 0.066, respectively, and were more promise than T2-Mapping mean (p<0.001). The AUC for differentiating AAR from the remote zone is 0.942±0.041. Texture features are not associated with convalescent decreased strain, ejection fraction (EF) or left ventricle remodeling (LVR) in analysis (p>0.05). CONCLUSION: TA of T2-mapping can distinguish AAR from both the infarct zone and the remote myocardial zone without LGE imaging in reperfused AMI. However, these features are not able to predict patients' functional recovery in the convalescent stage.

3.
Biotechnol Bioeng ; 118(8): 3105-3116, 2021 08.
Artigo em Inglês | MEDLINE | ID: mdl-34002369

RESUMO

Many microRNAs (miRNAs) have been predicted from small RNA sequencing data, but little was experimentally verified due to the lack of effective methods. Here, we developed a simple and reliable dual gene expression cassette vector-based method to verify predicted plant miRNAs. We cloned osa-miR528 as a known miRNA, hvu-miRX as a predicted miRNA and TaDREB3 open reading frame as a non-miRNA into the first gene expression cassette and fused their complementary or noncomplementary sequences as predicted target or nontarget sequences with the 3' untranslated region of green fluorescent protein (GFP) in the second one. When these constructs were bombarded into plant cells, only the construct containing both osa-miR528 or hvu-miRX and its complementary sequence did not generate green fluorescence. Stem-loop reverse-transcription polymerase chain reaction detected mature osa-miR528 or mature hvu-miRX in the cells, while northern analysis showed that GFP messenger RNA from the construct containing both osa-miR528 or hvu-miRX and its complementary sequence was degraded. Taken together, the results indicate that hvu-miRX is an authentic miRNA like osa-miR528, miRNA's complementary sequence is its target sequence, and both osa-miR528 and hvu-miRX silenced the GFP expression via a cleavage mode. Our method thus facilitates the verification of predicted plant miRNAs, target sequences, and function modes.

4.
Artigo em Inglês | MEDLINE | ID: mdl-32940092

RESUMO

OBJECTIVE: To investigate the effect of laparoscopic surgery in colorectal cancer (CRC) patients with natural orifice specimen extraction (NOSE) on the recovery and quality of life (QOL) of patients. MATERIAL AND METHODS: Ninety-two eligible patients were randomly assigned into two groups: the traditional laparoscopy group (L group, n = 46) and the laparoscopic transanal specimen extraction group (NL group, n = 46). General data, surgery-related indicators, postoperative recovery, and prognosis were compared and analyzed between the two groups. RESULTS: A total of 46 patients in each group were enrolled in this study. The general data and surgery-related indicators were comparable between the two groups (all p > .05). There were no significant differences in the time of first flatus, bleeding, obstruction, constipation, and infectious complications between the two groups (all p > .05). The differences in the incidence of postoperative diarrhea, pain degree, and satisfaction on the aesthetics of the abdominal wall showed significant differences (χ2 = 6.133, p = .013; χ2 = 12.116, p = .017; χ2 = 13.463, p = .004). The postoperative follow-up time was 3-53 months. There were no significant differences in the postoperative hospital stay, medical costs, hospital readmission rate, incidence of incisional hernia, overall survival, disease-free survival, and QOL between the two groups (all p > .05). Conclusion: Laparoscopic surgery with NOSE for eligible patients with CRC was a feasible choice.

5.
ACS Appl Mater Interfaces ; 12(32): 36259-36267, 2020 Aug 12.
Artigo em Inglês | MEDLINE | ID: mdl-32667180

RESUMO

As appealing alternatives to noble-metal-based electrocatalysts for catalyzing hydrogen evolution reaction (HER) in alkali electrolyzers, earth-abundant MoNi-based catalysts have attracted intensive attention. Unfortunately, the exploration of MoNi-based electrocatalysts with superior intrinsic activity and ultralong-term stability still remains a grand challenge. Here, ultralong high-index faceted Mo@MoNi core-shell nanowires were topochemically synthesized through the thermal reduction of Mo@NiMoO4 core-shell nanowires, where single-crystalline Mo support facilitates the topological transformation of NiMoO4 into high-index faceted MoNi. When the as-achieved Mo@MoNi core-shell nanowire film serve as a free-standing cathode in alkaline solutions, it exhibit a remarkably decreased HER overpotential of 18 mV at 10 mA cm-2 and a Tafel slope of ∼33 mV dec-1, which are much lower than those for the state-of-the-art earth-abundant electrocatalysts and even commercial Pt/C. Experimental and theoretical investigations reveal that the exposed high-index (331) facets of MoNi can considerably reduce the energy barriers of initial water dissociation and subsequent hydrogen combination steps, which synergistically accelerates the sluggish alkaline HER kinetics. Significantly, after a 70-day HER operation, the overpotential of Mo@MoNi electrocatalysts at 10 mA cm-2 decreases by only 4 mV. Therefore, this work sheds a bright light on the rational design of high-performance HER electrocatalysts and their practical utilization for alkaline electrolyzers.

6.
Nan Fang Yi Ke Da Xue Xue Bao ; 40(2): 147-151, 2020 Feb 29.
Artigo em Chinês | MEDLINE | ID: mdl-32376532

RESUMO

The SARS-CoV-2 epidemic starting in Wuhan in December, 2019 has spread rapidly throughout the nation. The control measures to contain the epidemic also produced influences on the transport and treatment process of patients with acute myocardial infarction (AMI), and adjustments in the management of the patients need to be made at this particular time. AMI is characterized by an acute onset with potentially fatal consequence, a short optimal treatment window, and frequent complications including respiratory infections and respiratory and circulatory failure, for which active on-site treatment is essential. To standardize the management and facilitate the diagnosis and treatment, we formulated the guidelines for the procedures and strategies for the diagnosis and treatment of AMI, which highlight 5 Key Principles, namely Nearby treatment, Safety protection, Priority of thrombolysis, Transport to designated hospitals, and Remote consultation. For AMI patients, different treatment strategies are selected based on the screening results of SARS-CoV-2, the time window of STEMI onset, and the vital signs of the patients. During this special period, the cardiologists, including the interventional physicians, should be fully aware of the indications and contraindications of thrombolysis. In the transport and treatment of AMI patients, the physicians should strictly observe the indications for patient transport with appropriate protective measurements of the medical staff.


Assuntos
Infecções por Coronavirus , Infarto do Miocárdio , Pandemias , Pneumonia Viral , Betacoronavirus , COVID-19 , Consenso , Infecções por Coronavirus/complicações , Infecções por Coronavirus/epidemiologia , Infecções por Coronavirus/prevenção & controle , Humanos , Infarto do Miocárdio/diagnóstico , Infarto do Miocárdio/terapia , Pandemias/prevenção & controle , Pneumonia Viral/complicações , Pneumonia Viral/epidemiologia , Pneumonia Viral/prevenção & controle , Guias de Prática Clínica como Assunto , Consulta Remota , SARS-CoV-2 , Terapia Trombolítica , Transporte de Pacientes
7.
Sci Rep ; 9(1): 17984, 2019 11 29.
Artigo em Inglês | MEDLINE | ID: mdl-31784629

RESUMO

The objectives of the present study were to examine the dynamic changes in breast milk melatonin throughout the course of lactation and to explore factors associated with changes in melatonin concentrations and rhythms in both preterm and term breast milk. Breast milk was collected sequentially at 03:00, 09:00, 15:00, and 21:00 in one day. Melatonin was analyzed in 392 breast milk samples from 98 healthy nursing mothers at 0 to 30 days postpartum. In both preterm and term breast milk, the melatonin concentration presented a circadian rhythm with the acrophase at around 03:00. Subgroup analysis showed the peak melatonin concentrations differed significantly across lactation stages, with the highest concentration in the colostrum, followed by transitional and mature breast milk. At 03:00, preterm breast milk had a higher concentration of melatonin than term breast milk in the colostrum (28.67 pg/mL vs. 25.31 pg/mL, p < 0.022), transitional breast milk (24.70 pg/mL vs. 22.55 pg/mL), and mature breast milk (22.37 pg/mL vs. 20.12 pg /mL). Further studies are warranted for their roles and significance on melatonin in breast milk in nutrition and metabolism of neonates.


Assuntos
Fenômenos Fisiológicos da Nutrição do Lactente , Lactação/fisiologia , Melatonina/análise , Leite Humano/química , Nascimento Prematuro/fisiopatologia , Adulto , Variação Biológica Individual , Aleitamento Materno , Ritmo Circadiano/fisiologia , Feminino , Humanos , Recém-Nascido , Recém-Nascido Prematuro/fisiologia , Idade Materna , Melatonina/metabolismo , Leite Humano/metabolismo , Período Pós-Parto/fisiologia , Gravidez
8.
Arch Med Sci ; 15(4): 979-991, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31360192

RESUMO

Introduction: Pancreatic ductal adenocarcinoma (PDAC) is one of the deadliest malignancies. However, the molecular mechanisms underlying PDAC are still not completely understood. Circular RNAs (circRNAs) are a unique class of RNA formed by special loop splicing. More and more researchers have paid attention to circRNAs. Material and methods: In this study, we constructed a circRNA-mediated competing endogenous RNA (ceRNA) network in PDAC. Gene ontology (GO) analysis was performed to explore circRNAs' potential roles in PDAC progression. We also constructed an up-stream transcriptional network of circRNAs' parental genes and found that many transcription factors (TFs), such as tumor protein p53 (TP53) and MYC, could regulate their expression. Results: This study, which aimed to identify differentially expressed circRNAs in PDAC, suggested that circRNAs may also act as biomarkers for PDAC. We analyzed two public datasets (GSE69362 and GSE79634) to identify differentially expressed circRNAs in PDAC. Finally, we found that DExH-Box Helicase 9 (DHX9) may be a potential regulator of circRNA formation in PDAC. Genomic loci of four down-regulated circRNAs - hsa_circ_000691, hsa_circ_0049392, hsa_circ_0005203, and hsa_circ_0001626 - contained DHX9 binding sites, suggesting that they may be directly regulated by DHX9. Conclusions: Our study identified differentially expressed circRNAs in PDAC, suggesting that circRNAs may also act as biomarkers for PDAC. Additional investigations of function and up-stream regulation of differentially expressed circRNA in PDAC are still needed.

9.
ACS Appl Mater Interfaces ; 11(29): 25682-25690, 2019 Jul 24.
Artigo em Inglês | MEDLINE | ID: mdl-31246393

RESUMO

Protein imprinting technology is of interest in drug delivery, biosensing, solid-phase extraction, and so forth. However, the efficient recognition and separation of proteins have remained challenging to date. Toward this, under the assistance of Ni2+-bovine serum albumin (BSA) directional coordination strategy, magnetic BSA-imprinted materials had been synthesized via dopamine self-polymerization on hollow Fe3O4@mSiO2 microspheres (mSiO2 referred as mesoporous silica). The well-defined imprinted microspheres possessed more satisfactory adsorption capacity (266.99 mg/g), enhanced imprinting factor (5.45), and fast adsorption saturation kinetics (40 min) for BSA, superior to many previous reports. Benefiting from the coordinate interaction between Ni2+ and BSA, these fabricated microspheres exhibited excellent specificity not only in individual and competitive protein rebinding samples but also in bovine serum. Combined with the directional coordination method, the magnetic-imprinted composite materials to selectively capture target proteins could provide promising potential in applications.


Assuntos
Óxido Ferroso-Férrico/química , Microesferas , Impressão Molecular , Níquel/química , Soroalbumina Bovina/química , Dióxido de Silício/química , Adsorção , Animais , Bovinos , Humanos , Propriedades de Superfície
10.
Zhongguo Zhong Yao Za Zhi ; 44(5): 927-934, 2019 Mar.
Artigo em Chinês | MEDLINE | ID: mdl-30989851

RESUMO

The dynamic accumulation rule of active substances in medicinal plants is of great value not only for medicinal material production and application,but also for the genetic mechanism study on the formation of medicinal ingredients,especially vital to guide medicinal material collection as well as experiment material selection and candidate gene screening in the analysis of biosynthesis pathway. This study investigated the accumulation of curcumins and terpenoids,and the biosynthesis of these metabolites,which are the active metabolites in Curcuma longa,a commonly used traditional Chinese medicine. Rhizoma of C. longa from leaf growing period,rhizome swelling period and dry matter accumulating period were used as experimental materials,to analyze the changes of metabolites and biosynthesis in the three periods by comparative transcriptome and metabolomes analysis.The results indicated that terpenoids accumulation and biosynthesis mainly occurred in leaf growing period,while curcumin accumulation and biosynthesis mainly occurred in dry matter accumulating period. Therefore,we suggested that turmeric rhizomes in leaf growth period were suitable for terpenoids biosynthetic pathway characterization,and rhizome in accumulation of dry matter period was suitable for curcuminoid biosynthesis pathway characterization. This study provides references for medicinal materialproduction and application,as well as biopathway analysis of active compounds for C. longa.


Assuntos
Curcuma/química , Curcumina/análise , Rizoma/química , Terpenos/análise , Compostos Fitoquímicos/análise , Plantas Medicinais/química
11.
J Magn Reson Imaging ; 48(6): 1602-1609, 2018 12.
Artigo em Inglês | MEDLINE | ID: mdl-29734489

RESUMO

BACKGROUND: Acute myocardial infarction (AMI) is a major cause of morbidity and mortality worldwide and places a significant financial burden on our society. PURPOSE: To determine the feasibility of cardiac intravoxel incoherent motion (IVIM) in the consecutive evaluation of myocardial perfusion in myocardial infarction patients postpercutaneous coronary intervention (PCI) and to investigate the dynamic biological phenomena in myocardial perfusion after AMI. STUDY TYPE: Prospective observational study. POPULATION: Twenty ST-segment elevation myocardial infarction (STEMI) patients after reperfusion therapy and 12 healthy volunteers served as controls. FIELD STRENGTH/SEQUENCE: Cardiac MRI at 3T, including steady-state free precession (SSFP) cine imaging, T2 -short time inversion recovery (T2 -STIR), late gadolinium enhancement (LGE), T2 mapping, and IVIM diffusion-weighted imaging (DWI) were performed. ASSESSMENT: Myocardial T2 value and IVIM-DWI-associated parameters (ADCfast , ADCslow , and f value) of the infarcted myocardium at different timepoints, remote myocardium, and normal myocardium were analyzed by two experienced radiologists. STATISTICAL TESTS: Independent sample's t-test, Pearson's, and Spearman's correlation and interobserver variability were applied. P ≤ 0.05 was considered significant. RESULTS: The T2 value in ischemic myocardium measured on day 3 (73.58 ± 4.37 msec) was greater than at any other timepoint (24 hours, day 7, day 30; 66.66 ± 4.71 msec, 68.36 ± 4.18 msec, 64.98 ± 5.39 msec, respectively, P < 0.001). ADCfast and f values were significantly lower in ischemic myocardium than in the remote myocardium as well. The f value in ischemic myocardium at day 3 (0.0989 ± 0.02) was lower than at any other timepoint (24 hours, 7 day, 30 day; 0.1203 ± 0.02, 0.1109 ± 0.02, 0.1213 ± 0.02, respectively, P < 0.001. DATA CONCLUSION: This preliminary study demonstrated that a dynamic process exists in the status of myocardial edema and myocardial perfusion in MI patients after PCI. The findings suggest myocardial perfusion would be best evaluated between day 3 and day 7. LEVEL OF EVIDENCE: 2 Technical Efficacy: Stage 2 J. Magn. Reson. Imaging 2018;48:1602-1609.


Assuntos
Imagem de Difusão por Ressonância Magnética , Coração/diagnóstico por imagem , Infarto do Miocárdio/diagnóstico por imagem , Imagem de Perfusão do Miocárdio , Adulto , Idoso , Meios de Contraste , Edema , Feminino , Gadolínio , Humanos , Imagem Cinética por Ressonância Magnética , Masculino , Pessoa de Meia-Idade , Movimento (Física) , Isquemia Miocárdica/diagnóstico por imagem , Miocárdio/patologia , Intervenção Coronária Percutânea , Estudos Prospectivos
12.
J Clin Lab Anal ; 32(2)2018 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-28444906

RESUMO

BACKGROUND: Carbamoyl Phosphate Synthetase 1 deficiency (CPS1D) is a rare autosomal recessive inborn metabolic disease characterized mainly by hyperammonemia. The fatal nature of CPS1D and its similar symptoms with other urea cycle disorders (UCDs) make its diagnosis difficult, and the molecular diagnosis is hindered due to the large size of the causative gene CPS1. Therefore, the objective of the present study was to investigate the clinical applicability of exome sequencing in molecular diagnosis of CPS1D in Chinese population. METHODS: We described two Chinese neonates presented with unconsciousness and drowsiness due to deepening encephalopathy with hyperammonemia. Whole exome sequencing was performed. Candidate mutations were validated by Sanger sequencing. In-silicon analysis was processed for the pathogenicity predictions of the identified mutations. RESULTS: Two compound heterozygous mutations in the gene carbamoyl phosphate synthetase 1(CPS1) were identified. One is in Case 1 with two novel missense mutations (c.2537C>T, p. Pro846Leu and c.3443T>A, p.Met1148Lys), and the other one is in Case 2 with a novel missense mutation (c.1799G>A, p.Cys600Tyr) and a previously reported 12-bp deletion (c.4088_4099del, p.Leu 1363_Ile1366del). Bioinformatics deleterious predictions indicated pathogenicity of the missense mutations. Conversation analysis and homology modeling showed that the substituted amino acids were highly evolutionary conserved and necessary for enzyme stability or function. CONCLUSION: The present study initially and successfully applied whole exome sequencing to the molecular diagnosis of CPS1D in Chinese neonates, indicating its applicability in cost-effective molecular diagnosis of CPS1D. Three novel pathogenic missense mutations were identified, expanded the mutational spectrum of the CPS1 gene.


Assuntos
Carbamoil-Fosfato Sintase (Amônia)/genética , Doença da Deficiência da Carbamoil-Fosfato Sintase I/diagnóstico , Doença da Deficiência da Carbamoil-Fosfato Sintase I/genética , Técnicas de Diagnóstico Molecular/métodos , Sequenciamento Completo do Exoma/métodos , Feminino , Humanos , Recém-Nascido , Masculino , Modelos Moleculares , Mutação/genética
13.
BMC Genomics ; 17(1): 735, 2016 09 15.
Artigo em Inglês | MEDLINE | ID: mdl-27633252

RESUMO

BACKGROUND: Various small RNA (sRNA) sizes and varieties have been identified, but their relationship as well as relationship with their origins and allocations have not been well understood or investigated. RESULTS: By comparing sRNAs generated from two barley cultivars, Golden Promise (GP) and Pallas, we identified that the generation of different sizes and types of sRNAs in barley was locus-, chromosome- and/or cultivar-dependent. 20-nt sRNAs mainly comprising miRNAs and chloroplast-derived sRNAs were significantly over-expressed in Pallas vs. GP on chromosomes 3H and 6H. MiRNAs-enriched 21-nt sRNAs were significantly over-expressed in Pallas vs. GP only on chromosome 4H. On chromosome 5H this size of sRNAs was significantly under-expressed in Pallas, so were 22-nt sRNAs mainly comprising miRNAs and repeat-derived sRNAs. 24-nt sRNAs mostly derived from repeats were evenly distributed in all chromosomes and expressed similarly between GP and Pallas. Unlike other sizes of sRNAs, 24-nt sRNAs were little conserved in other plant species. Abundant sRNAs were mostly generated from 3' terminal regions of chromosome 1H and 5' terminal regions of chromosome 5H. Over-expressed miRNAs in GP vs. Pallas primarily function in stress responses and iron-binding. CONCLUSIONS: Our study indicates that 23-24-nt sRNAs may be linked to repressive chromatin modifications and function in genome stability while 20-21-nt sRNAs may be important for the cultivar specificity. This study provides a novel insight into the mechanism of sRNA expression and function in barley.


Assuntos
Cromossomos de Plantas , Loci Gênicos , Hordeum/genética , RNA de Plantas/genética , Pequeno RNA não Traduzido/genética , Evolução Molecular , Regulação da Expressão Gênica de Plantas , Genoma de Planta , Estudo de Associação Genômica Ampla , Genômica/métodos , MicroRNAs/genética
14.
Biochem J ; 473(21): 3855-3869, 2016 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-27555611

RESUMO

Forkhead transcription factors play a key role in embryonic patterning during development. In Saccharomyces cerevisiae, two forkhead transcription factors, Fkh1 and Fkh2, regulate the transcription of CLB2 cluster genes important for mitosis. Fkh1 reduces, whereas Fkh2 elevates, the transcription of CLB2 cluster genes. However, the mechanism for this observation remains unclear. Fkh1 and Fkh2 each contain a forkhead domain (DNA-binding domain, DBD) and a forkhead-associated domain (FHAD), whereas Fkh2 possesses an extra C' domain containing six consensus cyclin-dependent kinase phosphorylation sites. In the present study, roles of these domains in protein complexes, the regulation of cell growth and CLB2 cluster genes and protein interactions were investigated using various domain mutants. The result showed that the DBD was vital for ternary complex formation with Mcm1, whereas the FHAD was central for the regulation of cell growth and CLB2 cluster transcription and for interactions with Ndd1 and Clb2. However, the Fkh2 C' domain was dispensable for the above functions. Both DBDs and FHADs had functional divergences in the cell, and Ndd1 functioned via its phosphorylated form. These data provide important insights into the functional mechanism of Fkh1 and Fkh2 in cell cycle control.


Assuntos
Proteínas de Ciclo Celular/metabolismo , Fatores de Transcrição Forkhead/metabolismo , Proteínas de Saccharomyces cerevisiae/metabolismo , Northern Blotting , Western Blotting , Ciclo Celular/genética , Ciclo Celular/fisiologia , Proteínas de Ciclo Celular/genética , Ciclina B/genética , Ciclina B/metabolismo , Fatores de Transcrição Forkhead/genética , Imunoprecipitação , Proteína 1 de Manutenção de Minicromossomo/genética , Proteína 1 de Manutenção de Minicromossomo/metabolismo , Plasmídeos , Proteínas de Saccharomyces cerevisiae/genética , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo
16.
Asian Pac J Cancer Prev ; 16(14): 6011-7, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26320489

RESUMO

PURPOSE: To investigate the efficacy and safety of selective radiotherapy after distant metastasis of nasopharyngeal carcinoma (NPC) treated with dose-dense cisplatin plus fluorouracil. MATERIALS AND METHODS: Eligible patients were randomly assigned to a study group treated with dose-dense cisplatin plus fluorouracil following selective radiotherapy and a control group receiving traditional cisplatin plus fluorouracil following selective radiotherapy according to a 1:1 distribution using a digital random table method. The primary endpoint was overall survival (OS). Secondary endpoints were progression-free survival (PFS), objective response rate, relapse or progression rate in the radiation field and treatment toxicity. RESULTS: Of 52 patients in the study group, 20 cases underwent radiotherapy., while in the control group of 51 patients, 16 underwent radiotherapy. The median PFS, median OS, survival rates in 1, 2 and 3 years in study and control group were 20.9 vs 12.7months, 28.3 vs 18.8months, 85.2%vs 65.9%, 62.2% vs 18.3%, and 36.6%vs 5.2% (p values of 0.00, 0.00, 0.04, 0.00 and 0.00, respectively). Subgroup analysis showed that the median OS and survival rates of 1, 2, 3 years for patients undergoing radiotherapy in the study group better than that in control group( 43.2vs24.1 months, 94.1% vs 86.7%, 82.4% vs 43.3%, 64.7% vs 17.3%, (p=0.00, 0.57, 0.04 and 0.01, respectively). The complete response rate, objective response rate after chemotherapy and three months after radiotherapy, relapse or progression rate in radiation field in study group and in control group were 19.2% vs 3.9%, 86.5% vs 56.9%, 85% vs 50%, 95% vs 81.3% and 41.3% vs 66.7% (p =0.03, 0.00, 0.03,0.30, 0.01 respectively). The grade 3-4 acute adverse reactions in the study group were significantly higher than in the control group (53.8% vs 9.8%, p=0.00). CONCLUSIONS: The survival of patients benefits from selective radiotherapy after distant metastasis of NPC treated with dose-dense cisplatin plus fluorouracil.


Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Neoplasias Ósseas/terapia , Quimiorradioterapia , Neoplasias Hepáticas/terapia , Neoplasias Pulmonares/terapia , Neoplasias Nasofaríngeas/tratamento farmacológico , Adolescente , Adulto , Idoso , Neoplasias Ósseas/mortalidade , Neoplasias Ósseas/secundário , Carcinoma , Estudos de Casos e Controles , Cisplatino/administração & dosagem , Feminino , Fluoruracila/administração & dosagem , Seguimentos , Humanos , Neoplasias Hepáticas/mortalidade , Neoplasias Hepáticas/secundário , Neoplasias Pulmonares/mortalidade , Neoplasias Pulmonares/secundário , Metástase Linfática , Masculino , Pessoa de Meia-Idade , Carcinoma Nasofaríngeo , Neoplasias Nasofaríngeas/mortalidade , Neoplasias Nasofaríngeas/patologia , Estadiamento de Neoplasias , Prognóstico , Taxa de Sobrevida , Adulto Jovem
17.
Front Plant Sci ; 6: 281, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25964791

RESUMO

Viral satellite RNAs (satRNAs) are small subviral RNAs and depend on the helper virus for replication and spread. satRNAs can attenuate helper virus-induced symptoms, the mechanism of which remains unclear. Here, we show that two virus-encoded suppressors of RNA silencing (VSRs), Cucumber mosaic virus (CMV) 2b and Tombusvirus P19, suppress hairpin RNA (hpRNA)-induced silencing of a ß-glucuronidase (GUS) gene in Nicotiana benthamiana. This suppression can be overcome by CMV Y-satellite RNA (Y-Sat) via the Y-Sat-derived small interfering RNAs (siRNAs), which bind to the VSRs and displace the bound hpGUS-derived siRNAs. We also show that microRNA target gene expression in N. tabacum was elevated by CMV infection, presumably due to function of the 2b VSR, but this upregulation of microRNA target genes was reversed in the presence of Y-Sat. These results suggest that satRNA infection minimizes the effect of VSRs on host siRNA and microRNA-directed silencing. Our results suggest that the high abundance of satRNA-derived siRNAs contributes to symptom attenuation by binding helper virus-encoded VSRs, minimizing the capacity of the VSRs to bind host siRNA and miRNA and interfere with their function.

18.
J Gastrointest Surg ; 19(8): 1391-8, 2015 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-25943912

RESUMO

BACKGROUND: The aim of this study was to investigate the role of the fast-track surgery (FTS) program in elderly patients (aged ≥75 years) who underwent open surgery for gastric cancer (GC) in China. METHODS: A total of 256 patients with GC were randomly assigned to four groups, each of which consisted of 64 cases: the 45-74-year-old age group was subdivided into the FTS-1 group and the conventional care (CC)-1 group, and the 75-89-year-old age group was subdivided into the FTS-2 group and the CC-2 group. All patients underwent open gastrectomy by the same experienced surgical team. We compared the differences between the pairs of groups in different age ranges with respect to the postoperative recovery index, complications, and medical costs. RESULTS: Compared with the CC-1 group, the FTS-1 group exhibited earlier postoperative flatus, a shorter postoperative hospital stay, lower medical costs, and a decreased incidence of sore throat (P = 0.010, P = 0.000, P = 0.000, and P = 0.019, respectively). Compared with the CC-2 group, the FTS-2 group had more nausea and vomiting, stomach retention, and intestinal obstruction, as well as a higher readmission rate (P = 0.015, P = 0.011, P = 0.041, and P = 0.013, respectively). CONCLUSION: The application of FTS can significantly speed up postoperative rehabilitation, shorten the hospitalization time, and lower the medical costs for 45-74-year-old GC patients, but this procedure does not show the same benefits for elderly patients. These findings suggest that we should carefully consider whether the FTS program should be applied to elderly patients with GC.


Assuntos
Gastrectomia , Obstrução Intestinal/etiologia , Assistência Perioperatória/métodos , Neoplasias Gástricas/cirurgia , Idoso , Idoso de 80 Anos ou mais , China , Deambulação Precoce , Estudos de Viabilidade , Feminino , Gastrectomia/efeitos adversos , Esvaziamento Gástrico , Custos de Cuidados de Saúde , Humanos , Tempo de Internação/economia , Masculino , Pessoa de Meia-Idade , Náusea/etiologia , Readmissão do Paciente , Faringite/etiologia , Recuperação de Função Fisiológica , Resultado do Tratamento , Vômito/etiologia
19.
PLoS One ; 10(3): e0118503, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25793505

RESUMO

For accurate and reliable gene expression analysis using quantitative real-time reverse transcription PCR (qPCR), the selection of appropriate reference genes as an internal control for normalization is crucial. We hypothesized that non-coding, small nucleolar RNAs (snoRNAs)would be stably expressed in different barley varieties and under different experimental treatments,in different tissues and at different developmental stages of plant growth and therefore might prove to be suitable reference genes for expression analysis of both microRNAs (miRNAs)and mRNAs. In this study, we examined the expression stability of ten candidate reference genes in six barley genotypes under five experimental stresses, drought, fungal infection,boron toxicity, nutrient deficiency and salinity. We compared four commonly used housekeeping genes; Actin (ACT), alpha-Tubulin (α-TUB), Glycolytic glyceraldehyde-3-phosphate dehydrogenase(GAPDH), ADP-ribosylation factor 1-like protein (ADP), four snoRNAs; (U18,U61, snoR14 and snoR23) and two microRNAs (miR168, miR159) as candidate reference genes. We found that ADP, snoR14 and snoR23 were ranked as the best of these candidates across diverse samples. Additionally, we found that miR168 was a suitable reference gene for expression analysis in barley. Finally, we validated the performance of our stable and unstable candidate reference genes for both mRNA and miRNA qPCR data normalization under different stress conditions and demonstrated the superiority of the stable candidates. Our data demonstrate the suitability of barley snoRNAs and miRNAs as potential reference genes form iRNA and mRNA qPCR data normalization under different stress treatments [corrected].


Assuntos
Regulação da Expressão Gênica de Plantas , Genes de Plantas , Hordeum/genética , Hordeum/fisiologia , MicroRNAs/genética , Estresse Fisiológico/genética , MicroRNAs/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase em Tempo Real/normas , Padrões de Referência , Reprodutibilidade dos Testes , Software
20.
Plant Biotechnol J ; 13(3): 293-305, 2015 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-25583362

RESUMO

Drought is a normal and recurring climate feature in most parts of the world and plays a major role in limiting crop productivity. However, plants have their own defence systems to cope with adverse climatic conditions. One of these defence mechanisms is the reprogramming of gene expression by microRNAs (miRNAs). miRNAs are small noncoding RNAs of approximately 22 nucleotides length, which have emerged as important regulators of genes at post-transcriptional levels in a range of organisms. Some miRNAs are functionally conserved across plant species and are regulated by drought stress. These properties suggest that miRNA-based genetic modifications have the potential to enhance drought tolerance in cereal crops. This review summarizes the current understanding of the regulatory mechanisms of plant miRNAs, involvement of plant miRNAs in drought stress responses in barley (Hordeum vulgare L.), wheat (Triticum spp.) and other plant species, and the involvement of miRNAs in plant-adaptive mechanisms under drought stress. Potential strategies and directions for future miRNA research and the utilization of miRNAs in the improvement of cereal crops for drought tolerance are also discussed.


Assuntos
Regulação da Expressão Gênica de Plantas , Hordeum/genética , MicroRNAs/genética , Plantas/genética , Triticum/genética , Secas , RNA de Plantas/genética , Estresse Fisiológico
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