RESUMO
Adipose tissue plays an important role in energy metabolism. Adipose dysfunction is closely related to obesity and type II diabetes. Glucose uptake is the key step for fat synthesis in adipocyte. miRNAs have been proven to play a crucial role in adipocyte differentiation, adipogenesis and glucose homeostasis. In this paper, we firstly reported that miR-146b decreased glucose consumption by up-regulating miR-146b in a porcine primary adipocyte model, while the inhibitor of endogenous miR-146b rescued the reduction. Then, miR-146b was predicated to target IRS1 by bioinformatics analysis, and a dual-luciferase reporter assay validated this predication. Western blot analyses indicated both IRS1 and glucose transporter type 4 (GLUT4) were down-regulated by miR-146b overexpression. Our study demonstrated that miR-146b regulated glucose homeostasis in porcine primary pre-adipocyte by targeting IRS1, and provided new understandings on regulations of lipogenesis by miRNAs.
Assuntos
Adipócitos/metabolismo , Glucose/metabolismo , Proteínas Substratos do Receptor de Insulina/metabolismo , MicroRNAs/metabolismo , Suínos/metabolismo , Adipogenia/genética , Tecido Adiposo , Animais , Sequência de Bases , Transportador de Glucose Tipo 4/genética , Transportador de Glucose Tipo 4/metabolismo , Proteínas Substratos do Receptor de Insulina/genética , Lipogênese/genética , Cultura Primária de Células , Suínos/genética , Regulação para CimaRESUMO
OBJECTIVE: A PCR-reverse dot blot hybridization (RDBH) assay was developed for rapid detection of rpoB gene mutations in 'hot mutation region' of Mycobacterium tuberculosis (M. tuberculosis). METHODS: 12 oligonucleotide probes based on the wild-type and mutant genotype rpoB sequences of M. tuberculosis were designed to screen the most frequent wild-type and mutant genotypes for diagnosing RIF resistance. 300 M. tuberculosis clinical isolates were detected by RDBH, conventional drug-susceptibility testing (DST) and DNA sequencing to evaluate the RDBH assay. RESULTS: The sensitivity and specificity of the RDBH assay were 91.2% (165/181) and 98.3% (117/119), respectively, as compared to DST. When compared with DNA sequencing, the accuracy, positive predictive value (PPV) and negative predictive value (NPV) of the RDBH assay were 97.7% (293/300), 98.2% (164/167), and 97.0% (129/133), respectively. Furthermore, the results indicated that the most common mutations were in codons 531 (48.6%), 526 (25.4%), 516 (8.8%), and 511 (6.6%), and the combinative mutation rate was 15 (8.3%). One and two strains of insertion and deletion were found among all strains, respectively. CONCLUSION: Our findings demonstrate that the RDBH assay is a rapid, simple and sensitive method for diagnosing RIF-resistant tuberculosis.
Assuntos
Antituberculosos/farmacologia , Farmacorresistência Bacteriana , Immunoblotting/métodos , Mycobacterium tuberculosis/efeitos dos fármacos , Rifampina/farmacologia , Genótipo , Testes de Sensibilidade Microbiana , Mycobacterium tuberculosis/genética , Reação em Cadeia da Polimerase/métodos , Sensibilidade e Especificidade , Fatores de TempoRESUMO
BACKGROUND: Ophthalmic ambulatory surgery is preferred to be performed under general anesthesia either by total intravenous anesthesia (TIVA) or by inhalational anesthesia to increase the patient comfort. However, anesthesia-controlled time (ACT) can cause increased non-operative operating room (OR) time which may adversely affect the ORs efficiency. This study was aimed to compare the ACT of desflurane with that of propofol-remifentanil in strabismus ambulatory surgery. METHODS: From November 2016 to December 2017, a total of 200 strabismus patients (aged 18-60 years old, and scheduled for elective ambulatory surgery at Zhongshan Ophthalmic Center) were randomly assigned to receive either propofol-based TIVA (group TIVA) or desflurane anesthesia (group DES) for maintenance of anesthesia. The primary outcome was the extubation time. Secondary outcomes included surgical time, anesthetic time, OR exit time, and Phase I and II recovery time. The intraoperative incidences of hypotension, bradycardia and oculocardiac reflex (OCR), and the incidences of any post-operative complications were recorded. Mann-Whitney U test and Chi-square or Fisher exact tests were used to compare the two groups. RESULTS: We found that the extubation time (5.5 [3.9-7.0] vs. 9.7 [8.5-11.4] min, Pâ<â0.001) and the incidence of prolonged time to extubation (0 vs. 6%, Pâ=â0.029) in the DES group were significantly decreased compared with those in the TIVA group. The patients in the DES group displayed shorter OR exit time as compared with that in the TIVA group (7.3 [5.5-8.7] vs. 10.8 [9.3-12.3] min, Pâ<â0.001). The patients using desflurane exhibited more stable hemodynamics during surgery than the patients using propofol-based TIVA, as demonstrated by lower incidences of hypotension (1% vs. 22%, Pâ<â0.001), bradycardia (2% vs. 13%, Pâ=â0.002), and OCR (17% vs. 44%, Pâ<â0.001). CONCLUSION: DES enhanced the ophthalmic OR efficiency by reducing the extubation time and OR exit time, and provided more stable hemodynamics intra-operatively than TIVA in patients undergoing strabismus ambulatory surgery. TRIAL REGISTRATION: ClinicalTrials.gov, No. NCT02922660; https://clinicaltrials.gov/ct2/show/NCT02922660?id=NCT02922660&draw=2&rank=1.
Assuntos
Anestesia Intravenosa/métodos , Desflurano/uso terapêutico , Estrabismo/cirurgia , Adolescente , Adulto , Procedimentos Cirúrgicos Ambulatórios/métodos , Anestesia Geral/métodos , Anestésicos Inalatórios/uso terapêutico , Anestésicos Intravenosos/uso terapêutico , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Salas Cirúrgicas , Duração da Cirurgia , Propofol/uso terapêutico , Remifentanil/uso terapêutico , Adulto JovemAssuntos
Carcinoma de Células Escamosas , Histerectomia , Terapia Neoadjuvante/métodos , Neoplasias do Colo do Útero , Adenocarcinoma/tratamento farmacológico , Adenocarcinoma/imunologia , Adenocarcinoma/patologia , Adenocarcinoma/radioterapia , Adenocarcinoma/cirurgia , Adulto , Antígenos de Neoplasias/metabolismo , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Carboplatina/administração & dosagem , Carcinoma de Células Escamosas/tratamento farmacológico , Carcinoma de Células Escamosas/imunologia , Carcinoma de Células Escamosas/patologia , Carcinoma de Células Escamosas/radioterapia , Carcinoma de Células Escamosas/cirurgia , Quimiorradioterapia Adjuvante , Quimioterapia Adjuvante , Feminino , Humanos , Radioisótopos de Irídio/uso terapêutico , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Paclitaxel/administração & dosagem , Período Pré-Operatório , Radioterapia Adjuvante , Estudos Retrospectivos , Serpinas/metabolismo , Resultado do Tratamento , Neoplasias do Colo do Útero/tratamento farmacológico , Neoplasias do Colo do Útero/imunologia , Neoplasias do Colo do Útero/patologia , Neoplasias do Colo do Útero/radioterapia , Neoplasias do Colo do Útero/cirurgiaRESUMO
Ammonia detoxification, which takes place via the hepatic urea cycle, is essential for nitrogen homeostasis and physiological well-being. It has been reported that a reduction in dietary protein reduces urea nitrogen. MicroRNAs (miRNAs) are major regulatory non-coding RNAs that have significant effects on several metabolic pathways; however, little is known on whether miRNAs regulate hepatic urea synthesis. The objective of this study was to assess the miRNA expression profile in a low protein diet and identify miRNAs involved in the regulation of the hepatic urea cycle using a porcine model. Weaned 28-days old piglets were fed a corn-soybean normal protein diet (NP) or a corn-soybean low protein diet (LP) for 30 d. Hepatic and blood samples were collected, and the miRNA expression profile was assessed by sequencing and qRT-PCR. Furthermore, we evaluated the possible role of miR-19b in urea synthesis regulation. There were 25 differentially expressed miRNAs between the NP and LP groups. Six of these miRNAs were predicted to be involved in urea cycle metabolism. MiR-19b negatively regulated urea synthesis by targeting SIRT5, which is a positive regulator of CPS1, the rate limiting enzyme in the urea cycle. Our study presented a novel explanation of ureagenesis regulation by miRNAs.
RESUMO
The relax circle DNA (rcDNA) sequence and the covalently closed circle DNA (cccDNA) sequence in hepatitis B virus (HBV) are crucial regions for HBV infections. To analyze mutations in rcDNA and cccDNA, DNA sequencing is often used, although it is time-consuming and expensive. Herein, we report a simple, economic, albeit accurate allele-specific polymerase chain reaction (AS-PCR) to detect mutations in these regions of HBV. This method can be extensively used to screen for mutations at specific positions of HBV genome.
Assuntos
DNA Circular/genética , DNA Viral/genética , Vírus da Hepatite B/genética , Mutação Puntual , Reação em Cadeia da Polimerase/métodos , Virologia/métodos , Alelos , Custos e Análise de Custo , Programas de Rastreamento/métodos , Reação em Cadeia da Polimerase/economia , Sensibilidade e Especificidade , Virologia/economiaRESUMO
Based on the "pressure-state-response (PSR)" concept model, a degradation evaluation index system was constructed for the cropland, wetland, and inshore ecosystems in Dongtan coastal zone of Chongming. By using multiplication synthesis, a combination of analytic hierarchy process and entropy weight method, the weights of each evaluation index were obtained, and, through geographical space index quantification and spatial clustering, the degradation degree of each ecological system was analyzed. The results showed that the degradation degree of Dongtan coastal ecosystems in 2005 could be spatially classified into four classes, i.e., class I, class II, class III and class IV, with the degradation degree aggravated increasingly. For the cropland, wetland, and inshore ecosystems, the weight of heavy metals was the largest, being 0.65, 0.20, and 0.26, respectively. Bird diversity index, land use degree, and Spartina alterniflora coverage also had greater effects on wetland ecosystem, and their weights were 0.26, 0.16, and 0.10, respectively. For cropland ecosystem, land use degree was also an important affecting factor, with the weight of 0.22.
Assuntos
Conservação dos Recursos Naturais/estatística & dados numéricos , Ecossistema , Monitoramento Ambiental/métodos , Áreas Alagadas , China , Oceanos e MaresRESUMO
Olfactory ensheathing cells (OECs) are a unique type of glial cells that have axonal growth-promoting properties. OEC transplantation has emerged as a promising experimental therapy of axonal injuries and demyelinating diseases. However, some fundamental cellular properties of OECs remain unclear. In this study, we found that the distinct OEC subpopulations exhibited different migratory properties based on time-lapse imaging of single isolated cells, possibly due to their different cytoskeletal organizations. Moreover, OEC subpopulations displayed different attractive migratory responses to a gradient of lysophosphatidic acid (LPA) in single-cell migration assays. Finally, we found that OEC subpopulations transformed into each other spontaneously. Together, these results demonstrate, for the first time to our knowledge, that distinct OEC subpopulations display different migratory properties in vitro and provide new evidence to support the notion of OECs as a single cell type with malleable functional phenotypes.
Assuntos
Bioensaio/métodos , Movimento Celular , Bulbo Olfatório/citologia , Animais , Linhagem Celular Transformada , Movimento Celular/efeitos dos fármacos , Células Cultivadas , Citoesqueleto/efeitos dos fármacos , Laminina/metabolismo , Lisofosfolipídeos/farmacologia , Masculino , Bulbo Olfatório/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Células de Schwann/citologia , Células de Schwann/efeitos dos fármacosRESUMO
Transplantation of Schwann cells (SCs) and olfactory ensheathing cells (OECs) have emerged as very promising therapies for spinal cord repair. The important features of interaction between SCs and OECs are beginning to be appreciated, although the underlying mechanism remains unclear. In the present study, we tested the effects of OECs on SCs migration using a range of in vitro migration assays. We found that SCs migrated abundantly upon OECs monolayer, and the migration-promoting effects were identified to be due to the secreted diffusible factors in OEC-derived conditioned medium (OEC-CM). Furthermore, neutralizing nerve growth factor (NGF) in OEC-CM with NGF antibody could block this effect. Moreover, we found that NGF promotes SCs migration even on astrocyte monolayer. Taken together, these findings provide the first evidence that OECs can promote SCs migration in astrocytic environment by secreted NGF.