RESUMO
The forkhead box F2 (Foxf2) gene suppresses epithelial-mesenchymal transition via the modulation of transcription of zinc finger E-box-binding homeobox 1 (Zeb1) and epithelial (E)-cadherin, thereby inhibiting tumor metastasis. Additionally, the specific binding of microRNA (miR)-200c to Foxf2 mRNA impedes metastatic pulmonary cancer. However, the role of miR-200c in breast cancer is still unknown. Therefore, in this study, miR-200c mimics were transfected into the highly metastatic breast cancer cell line MDA-MB-231. Their invasion and migration abilities were observed by scratch and transwell migration assays. Real-time PCR was used to detect mRNA levels of Foxf2, Zeb1, and E-cadherin, whereas Foxf2 protein level was determined by western blot analysis. Our results showed that, compared to the control group, miR-200c inhibited the migration or invasion of MDA-MB-231 cells. Real-time PCR and western blot analysis exhibited significant decreases in Foxf2 expression in the presence of miR-200c, along with a decrease in Zeb1 and increase in E-cadherin mRNA expressions. Thus, our preliminary data demonstrated that miR-200c could inhibit the metastasis of breast cancer cells by downregulating Foxf2 expression, providing leads for the discovery of a novel breast cancer treatment.
Assuntos
Neoplasias da Mama/metabolismo , Movimento Celular , Fatores de Transcrição Forkhead/genética , Regulação Neoplásica da Expressão Gênica , MicroRNAs/genética , Neoplasias da Mama/genética , Caderinas/genética , Caderinas/metabolismo , Linhagem Celular Tumoral , Regulação para Baixo , Feminino , Fatores de Transcrição Forkhead/metabolismo , Humanos , MicroRNAs/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Homeobox 1 de Ligação a E-box em Dedo de Zinco/genética , Homeobox 1 de Ligação a E-box em Dedo de Zinco/metabolismoRESUMO
BACKGROUND: This study evaluated the morphology of the rat liver when hyperbaric oxygen (HBO) was used at various stages of ischemia and reperfusion. METHODS: Thirty-two male Wistar rats, subjected to 30 minutes of hepatic ischemia and 30 minutes of reperfusion, were randomly assigned as follows: GIR (n = 8), control without HBO; GHBO/I (n = 8), in which HBO was applied only during ischemia; GHBO/R (n = 8), HBO only during reperfusion; and GHBO/IR (n = 8), HBO during both ischemia and reperfusion. Feasibility scores of hepatocytes were determined by assessing 8 items related to liver injury. RESULTS: The histologic injury score of the hepatic specimens was significantly lower in the GHBO/I group (79.0 ± 0.1) compared with the GIR group (135.0 ± 0.1). HBO was not effective when applied during reperfusion (GHBO/R, 151.3 ± 0.1) or during the ischemia plus reperfusion period (GHBO/IR, 131.0 ± 0.1). The sum was significantly higher (P < .05) in HBO-treated animals during the reperfusion period (ie, in the GHBO/R group compared with any of the other groups). CONCLUSIONS: A favorable effect was obtained when HBO was administered early during ischemia. HBO given in later periods of reperfusion was associated with a more severe sum index percentage of liver damage.
Assuntos
Oxigenoterapia Hiperbárica/métodos , Hepatopatias/terapia , Fígado/irrigação sanguínea , Oxigênio/metabolismo , Traumatismo por Reperfusão/terapia , Animais , Modelos Animais de Doenças , Precondicionamento Isquêmico , Fígado/patologia , Hepatopatias/metabolismo , Hepatopatias/patologia , Masculino , Ratos , Ratos Wistar , Traumatismo por Reperfusão/metabolismo , Traumatismo por Reperfusão/patologiaRESUMO
Tibetan sheep, an indigenous breed, have a wide variety of phenotypes and a colorful coat, which make this breed an interesting model for evaluating the effects of coat-color gene mutations on this phenotypic trait. The agouti signaling protein (ASIP) gene is a positional candidate gene, as was inferred based on previous study. In our research, ASIP gene copy numbers in genomic DNA were detected using a novel approach, and the exon 2 g.100-104 mutation and copy number variation (CNV) of ASIP were associated with coat color in 256 sheep collected from eight populations with different coat colors by high-resolution melting curve assay. We found that the relative copy numbers of ASIP ranged from one to eight in Tibetan sheep. All of the g.100-104 genotypes in the populations were in Hardy-Weinberg equilibrium, and there was no relationship between the g.100-104 genotype and coat color (P > 0.05). The single ASIP CNV allele was found to be almost entirely associated with solid-black coat color; however, not all solid-black sheep displayed the putative single ASIP CNV genotype. From our study, we speculate that the ASIP CNV is under great selective pressure and the single ASIP CNV allows selection for black coat color in Tibetan sheep, but this does not explain all black phenotypes in Tibetan sheep.
Assuntos
Proteína Agouti Sinalizadora/genética , Proteína Agouti Sinalizadora/fisiologia , Cabelo/fisiologia , Pigmentação , Carneiro Doméstico/genética , Alelos , Animais , Cor , DNA/genética , Variações do Número de Cópias de DNA , Éxons , Genômica , Genótipo , Íntrons , Mutação , Fenótipo , Polimorfismo de Nucleotídeo Único , TibetRESUMO
Wool is produced via synthetic processes of wool follicles, which are embedded in the skin of sheep. The development of new-generation sequencing and RNA sequencing provides new approaches that may elucidate the molecular regulation mechanism of wool follicle development and facilitate enhanced selection for wool traits through gene-assisted selection or targeted gene manipulation. We performed de novo transcriptome sequencing of skin using the Illumina Hiseq 2000 sequencing system in sheep (Ovis aries). Transcriptome de novo assembly was carried out via short-read assembly programs, including SOAPdenovo and ESTScan. The protein function, clusters of orthologous group function, gene ontology function, metabolic pathway analysis, and protein coding region prediction of unigenes were annotated by BLASTx, BLAST2GO, and ESTScan. More than 26,266,670 clean reads were collected and assembled into 79,741 unigene sequences, with a final assembly length of 35,447,962 nucleotides. A total of 22,164 unigenes were annotated, accounting for 36.27% of the total number of unigenes, which were divided into 25 classes belonging to 218 signaling pathways. Among them, there were 17 signal paths related to hair follicle development. Based on mass sequencing data of sheepskin obtained by RNA-Seq, many unigenes were identified and annotated, which provides an excellent platform for future sheep genetic and functional genomic research. The data could be used for improving wool quality and as a model for human hair follicle development or disease prevention.
Assuntos
Carneiro Doméstico/genética , Pele/metabolismo , Transcriptoma , Lã/fisiologia , Animais , Mapeamento de Sequências Contíguas , Etiquetas de Sequências Expressas , Perfilação da Expressão Gênica , Regulação da Expressão Gênica , Biblioteca Gênica , Genômica , Modelos Genéticos , Anotação de Sequência Molecular , Análise de Sequência de RNARESUMO
Invasion, metastasis, and recurrence are the most common causes of death in patients with hepatocellular carcinoma (HCC) and are therefore critical factors for both therapy and prognosis. Current methods for diagnosis of HCC rely mainly on serological markers such as alpha-fetoprotein and liver enzymes, together with physical assessment and imaging techniques. The availability of more accurate serum markers may facilitate screening and early diagnosis, which will improve prognosis. This retrospective cohort analysis included 50 consecutive patients with cirrhosis and single or multifocal HCC and 40 control subjects with no liver disease or risk factors for viral hepatitis. Expression of epidermal growth factor-like domain 7 (EGFL7), osteopontin (OPN), and prostaglandin E2 (PGE2) were detected using an enzyme-linked immunosorbent assay. The mean serum levels of EGFL7, OPN, and PGE2 in the HCC group were 132.11 pg/mL, 11.77 ng/mL, and 179.37 pg/mL, respectively, which were all significantly higher than the levels in the control group (23.03 pg/mL, 2.31 ng/mL, and 47.36 pg/mL, respectively; P < 0.001). Serum levels of EGFL7, OPN, and PGE2 levels may thus be useful for screening and surveillance of HCC among high-risk populations, and have the potential to improve prognosis of these patients.
Assuntos
Carcinoma Hepatocelular/sangue , Carcinoma Hepatocelular/genética , Regulação Neoplásica da Expressão Gênica , Neoplasias Hepáticas/sangue , Neoplasias Hepáticas/genética , Adulto , Idoso , Biomarcadores Tumorais/sangue , Proteínas de Ligação ao Cálcio , Dinoprostona/sangue , Família de Proteínas EGF , Fatores de Crescimento Endotelial/sangue , Ensaio de Imunoadsorção Enzimática , Feminino , Fibrose/sangue , Perfilação da Expressão Gênica , Marcadores Genéticos/genética , Humanos , Fígado/enzimologia , Masculino , Pessoa de Meia-Idade , Análise Multivariada , Metástase Neoplásica , Recidiva Local de Neoplasia , Osteopontina/sangue , Prognóstico , Estudos Retrospectivos , Fatores de Risco , alfa-Fetoproteínas/metabolismoRESUMO
This study investigated geographic and pairwise distances among seven Chinese local and four introduced sheep populations via analysis of 26 microsatellite DNA markers. Genetic polymorphism was rich, and the following was discovered: 348 alleles in total were detected, the average allele number was 13.38, the polymorphism information content (PIC) of loci ranged from 0.717 to 0.788, the number of effective alleles ranged from 7.046 to 7.489, and the observed heterozygosity ranged from 0.700 to 0.768 for the practical sample, and from 0.712 to 0.794 for expected heterozygosity. The Wright's F-statistic of subpopulations within the total (FST) was 0.128, the genetic differentiation coefficient (GST) was 0.115, and the average gene flow (Nm) was 1.703. The phylogenetic trees based on the neighbor-joining method by Nei's genetic distance (DA) and Nei's standard genetic distance (DS) were similar. Sheep populations clustered into group 1 (Ta, M, L, H, O, G, and Q breeds) and group 2 (PD, WS, B, and T breeds). These results will have an important value applied and directive significance for sheep breeding in the future.
Assuntos
Repetições de Microssatélites , Ovinos/genética , Animais , Evolução Molecular , Deriva Genética , Marcadores Genéticos , Variação Genética , Espécies Introduzidas , Filogeografia , Análise de Sequência de DNA , Ovinos/sangueRESUMO
Variation in microsatellite or simple sequence repeat (SSR) loci has, until recently, relied heavily on the use of gel-based methods that can be both time consuming and difficult to genotype. Non gel-based systems are therefore important to increase simplicity and improve turn-around time without compromising assay sensitivity and accuracy. In this report, we assessed the latest of the non-gel-based methods, high-resolution melting (HRM) curve analysis. HRM is a technique that monitors exactly the decreasing fluorescence of intercalating dye in the process of dissociation of double-stranded DNA. The measurement immediately follows polymerase chain reaction in a one-step, closed-tube method. Four SSR loci of different complexity in sheep, namely MAF209, MCM140, CB226, and SRCRSP5, were assessed using the LightScanners System with LC Greens PLUS DNA binding dye. In order to improve the accuracy of genotyping, we applied internal oligo nucleotide calibrators while performing HRM. DNA polymorphisms were previously identified using capillary electrophoresis analysis (CE). The result showed that CE detected more genotypes than HRM in the same loci regardless of the level of polymorphism at the SSR loci. We demonstrate current limitations of the HRM method for the analysis of SSR loci.
Assuntos
Técnicas de Genotipagem , Repetições de Microssatélites/genética , Carneiro Doméstico/genética , Animais , Eletroforese Capilar , Genótipo , Desnaturação de Ácido Nucleico/genéticaRESUMO
Endoluminal sonography is a technique well suited to imaging structures beyond the lumen of the hollow viscus. The development of small-diameter (6.2F), catheter-enclosed ultrasound probes has made this technique available for use within the urinary tract. It is capable of defining adjacent vessels, calculi, and masses. Ureteropelvic junction (UPJ) obstruction has been increasingly treated by incisional techniques, either nephroscopic, ureteroscopic, or radiographically controlled, with each incision at risk for causing damage to any adjacent vessel. Endoluminal sonography of the obstructed UPJ was attempted in 46 patients and completed in 45 patients, 41 with primary and 4 with secondary obstruction. Adjacent vessels could be seen in 24 patients. Twelve were located anterior or medial to the UPJ or both. Nine patients had vessels at the UPJ located laterally or anterolaterally, posterolaterally, or medially and laterally. Sonographic localization guided the choice of incision site in all patients and changed therapy in five patients. This technique also allows recognition of high insertion of the ureter into the renal pelvis. Endoluminal sonography of the obstructed UPJ is a valuable technique to determine the location and nature of associated vessels and, therefore, to guide decisions in treatment.
Assuntos
Ultrassonografia/métodos , Obstrução Ureteral/diagnóstico por imagem , Adulto , Angiografia , Artérias/diagnóstico por imagem , Desenho de Equipamento , Fluoroscopia , Humanos , Masculino , Ultrassonografia/instrumentação , Ureter/irrigação sanguínea , UrografiaRESUMO
During the past 25 years, many attempts have been made to establish effective ultrasound contrast agents for both cardiac and noncardiac applications. The ideal ultrasound contrast agent would be: (a) nontoxic; (b) injectable intravenously; (c) capable of passing through the pulmonary, cardiac and capillary circulations; and (d) stable for recirculation. A variety of potential ultrasound contrast agents have been or are now under development. Present and future ultrasound contrast agents should provide for increased diagnostic capabilities in a variety of normal and abnormal vessels and organs throughout the body. These agents will enhance tumor vascularity, delineate areas of ischemia, as well as improve visualization of vascular stenosis. Future developments with modification of ultrasound equipment should increase the capabilities of these agents to improve imaging as well as Doppler sensitivity.
Assuntos
Meios de Contraste , Ultrassonografia , Fenômenos Biofísicos , Biofísica , Meios de Contraste/administração & dosagem , Meios de Contraste/química , Meios de Contraste/farmacocinética , Ecocardiografia , HumanosRESUMO
Endoluminal sonography of the urinary tract was performed by using endoluminal ultrasound transducers contained within 2-mm-diameter catheters. The catheters were inserted into the urinary bladder via the urethra and advanced into the ureters and renal pelvis under cystoscopic control; then, cross-sectional images of the bladder, ureters, and renal pelvis were obtained. Two dogs and seven human patients were studied. In one dog, a 4.5-mm pseudopolyp, which was surgically created in the bladder wall, was successfully imaged; in the other, stones 2 mm or larger inserted into the bladder were identified. Of the seven patients, sonography showed stones embedded in the renal parenchyma (one patient) and the mucosa of the distal ureter (one patient). These were ultimately confirmed by their eventual removal. In a third, sonography showed a tumor of the distal ureter and identified the depth of the tumor. This was confirmed by biopsy. In a fourth, sonography clearly showed a crossing vessel as the cause for narrowing of the proximal ureter. In a fifth, sonography showed that the cause of a ureteral stricture was idiopathic. In the last two cases, sonography did not reveal a cause for hematuria. In these last three cases, negative sonographic results were confirmed by direct ureteroscopic examinations and follow-up studies. Our observations based on this limited study suggest that endoluminal sonography is a useful procedure for diagnosing diseases of the urinary tract. Further study is warranted.