RESUMEN
PURPOSE: This study aimed to investigate the efficacy and safety of ultrasound-guided percutaneous radiofrequency ablation (RFA) for the treatment of synovial hyperplasia in the knee joints of antigen-induced arthritis (AIA) model rabbits. METHODS: Forty Japanese large-eared white rabbits were divided into AIA and control groups. After successful induction of the AIA model, the knee joints were randomly assigned to RFA and non-RFA groups. The RFA group underwent ultrasound-guided RFA to treat synovial hyperplasia in the knee joint. Dynamic observation of various detection indices was conducted to evaluate the safety and effectiveness of the RFA procedure. RESULTS: Successful synovial ablation was achieved in the RFA group, with no intraoperative or perioperative mortality. Postoperative the circumference of the knee joint reached a peak before decreasing in the third week after surgery. The incidence and diameter of postoperative skin ulcers were not significantly different compared to the non-RFA group (p > .05). Anatomical examination revealed an intact intermuscular fascia around the ablated area in the RFA group. The ablated synovial tissue initially presented as a white mass, which subsequently liquefied into a milky white viscous fluid. Gross articular cartilage was observed, along with liquefied necrosis of the synovium on pathological histology and infiltration of inflammatory cells in the surrounding soft tissue. CONCLUSION: The experimental results demonstrated that ultrasound-guided RFA of the knee in the treatment of synovial hyperplasia in AIA model animals was both effective and safe.
Asunto(s)
Hiperplasia , Ablación por Radiofrecuencia , Animales , Conejos , Ablación por Radiofrecuencia/métodos , Hiperplasia/cirugía , Hiperplasia/patología , Membrana Sinovial/patología , Membrana Sinovial/diagnóstico por imagen , Ultrasonografía/métodos , Masculino , Ultrasonografía Intervencional/métodosRESUMEN
DNA is susceptible to various factors in vitro and in vivo and experience different forms of damage,among which double-strand break(DSB)is a deleterious form.To maintain the stability of genetic information,organisms have developed multiple mechanisms to repair DNA damage.Among these mechanisms,homologous recombination(HR)is praised for the high accuracy.The MRE11-RAD50-NBS1(MRN)complex plays an important role in HR and is conserved across different species.The knowledge on the MRN complex mainly came from the previous studies in Saccharomyces cerevisiae and Caenorhabditis elegans,while studies in the last decades have revealed the role of mammalian MRN complex in DNA repair of higher animals.In this review,we first introduces the MRN complex regarding the composition,structure,and roles in HR.In addition,we discuss the human diseases such as ataxia-telangiectasia-like disorder,Nijmegen breakage syndrome,and Nijmegen breakage syndrome-like disorder that are caused by dysfunctions in the MRN complex.Furthermore,we summarize the mouse models established to study the clinical phenotypes of the above diseases.
Asunto(s)
Ácido Anhídrido Hidrolasas , Proteínas de Ciclo Celular , Enzimas Reparadoras del ADN , Proteínas de Unión al ADN , Proteína Homóloga de MRE11 , Proteínas Nucleares , Humanos , Ácido Anhídrido Hidrolasas/metabolismo , Ácido Anhídrido Hidrolasas/genética , Proteína Homóloga de MRE11/metabolismo , Proteína Homóloga de MRE11/genética , Proteínas de Ciclo Celular/metabolismo , Proteínas de Ciclo Celular/genética , Proteínas de Unión al ADN/metabolismo , Proteínas de Unión al ADN/genética , Enzimas Reparadoras del ADN/metabolismo , Enzimas Reparadoras del ADN/genética , Proteínas Nucleares/metabolismo , Proteínas Nucleares/genética , Animales , Reparación del ADN , Ataxia Telangiectasia/genética , Ataxia Telangiectasia/metabolismo , Síndrome de Nijmegen/metabolismo , Síndrome de Nijmegen/genéticaRESUMEN
OBJECTIVE: To evaluate the relation between the expression of PD-1, PD-L1, CD3, CD8, Foxp3 and clinicopathological features in patients with oral leukoplakia (OLK) and oral squamous cell carcinomas (OSCC) as well as the malignant outcome in OLK patients, and to study the effect of PD-1 and PD-L1 on immune microenvironment in the progression of oral carcinogenesis. METHODS: We evaluated the expression of PD-1/PD-L1 and composition of CD3+ , CD8+ and Foxp3+ T lymphocytes in OLK and OSCC samples by immunohistochemical (IHC) staining and analyzed their relation with clinical information and malignant transformation in OLK patients. RESULTS: IHC staining demonstrated that the expression of PD-1 was significantly increased in the high-grade OLK group than in the low-grade OLK group, while PD-L1 was detected mainly in OSCC. The expression of CD3, CD8, and Foxp3 was found higher in the high-grade OLK group than in the low-grade OLK group, and the Foxp3+ cells were found more in the OSCC group than in the high-grade OLK group. PD-1 was significantly correlated with CD3 (p < 0.05, R = 0.52), CD8 (p < 0.05, R = 0.46), and Foxp3 (p < 0.05, R = 0.46), and the low PD-1-expression group showed a better malignant-free survival than high PD-1 expression group in the OLK (p < 0.05). CONCLUSION: The PD-1/PD-L1 may induce immune suppression in OLK and accelerate the progress of malignant transformation.
Asunto(s)
Carcinoma de Células Escamosas , Neoplasias de Cabeza y Cuello , Neoplasias de la Boca , Humanos , Neoplasias de la Boca/patología , Carcinoma de Células Escamosas de Cabeza y Cuello , Carcinoma de Células Escamosas/patología , Receptor de Muerte Celular Programada 1 , Antígeno B7-H1 , Leucoplasia Bucal/patología , Transformación Celular Neoplásica , Factores de Transcripción Forkhead , Microambiente TumoralRESUMEN
INTRODUCTION: The role of extra-hypothalamic thyrotropin-releasing hormone (TRH) has been investigated by pharmacological studies using TRH or its analogues and found to produce a wide array of effects in the central nervous system. METHODS: Immunofluorescence, In situ labeling of DNA (TUNEL), in situ hybridization chain reaction and quantitative real-time polymerase chain reaction were used in this study. RESULTS: We found that the granular cells of the dentate gyrus expressed transiently a significant amount of TRH-like immunoreactivity and TRH mRNA during the 6-24 h period following global cerebral ischemia/reperfusion injury. TUNEL showed that apoptosis of neurons in the CA1 region occurred from 48 h and almost disappeared at 7 days. TRH administration 30 min before or 24 h after the injury could partially inhibit neuronal loss, and improve the survival of neurons in the CA1 region. CONCLUSION: These data suggest that endogenous TRH expressed transiently in the dentate gyrus of the hippocampus may play an important role in the survival of neurons during the early stage of ischemia/reperfusion injury and that delayed application of TRH still produced neuroprotection. This delayed application of TRH has a promising therapeutic significance for clinical situations.
Asunto(s)
Isquemia Encefálica , Daño por Reperfusión , Animales , Isquemia Encefálica/metabolismo , Hipocampo/metabolismo , Péptidos/metabolismo , ARN Mensajero/metabolismo , Ratas , Daño por Reperfusión/metabolismo , Hormona Liberadora de Tirotropina/genética , Hormona Liberadora de Tirotropina/metabolismoRESUMEN
MicroRNAs (miRNAs) are a class of small noncoding RNAs that play important roles in plant growth and development as well as in stress responses. However, little is known about their regulatory functions affecting rice grain yield. We functionally characterized a novel miRNA in rice, OsmiR530, its target OsPL3, and its upstream regulator phytochrome-interacting factor-like 15 (OsPIL15). Their effects on rice yield were dissected comprehensively. We determined that OsmiR530 negatively regulates grain yield. Blocking OsmiR530 increases grain yield, whereas OsmiR530 overexpression significantly decreases grain size and panicle branching, leading to yield loss. Additionally, OsPL3, which encodes a PLUS3 domain-containing protein, is targeted directly by OsmiR530. Knocking out OsPL3 decreases the grain yield. In-depth analyses indicated that OsPIL15 activates OsMIR530 expression by directly binding to the G-box elements in the promoter. Analyses of genetic variations suggested that the OsMIR530 locus has likely been subjected to artificial selection during rice breeding. The results presented herein reveal a novel OsPIL15-OsmiR530 module controlling rice grain yield, thus providing researchers with a new target for the breeding of high-yielding rice.
Asunto(s)
Oryza , Fitocromo , Grano Comestible/metabolismo , Regulación de la Expresión Génica de las Plantas , Oryza/genética , Oryza/metabolismo , Fitocromo/metabolismo , Fitomejoramiento , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismoRESUMEN
The purpose of this study was to investigate the antimicrobial activity and mechanisms of Nα-lauroyl arginate ethyl ester (LAE) against Penicillium digitatum and Pectobacterium carotovorum subsp. carotovorum. The minim inhibitory concentrations of LAE against P. digitatum and P. carotovorum were found to be 400 and 25 µg/ml, respectively. Loss of intracellular protein and nucleic acid increased significantly, and membrane permeability reached 76.28, 54.29 and 85.20%, respectively, when 400 µg/ml of LAE was applied to the hyphae and spores of P. digitatum and to P. carotovorum. Flow cytometry showed that LAE reduced the membrane potential, and the depolarization ratios of P. digitatum and P. carotovorum were 98.19 and 97.25% (P < 0.05), respectively. Transmission electron microscopy photos revealed that LAE caused a rough surface, irregular cellular organelles, protoplast shrinkage, intracytoplasmic coagulation and empty cavities in all three cell types. These results showed that LAE had notable ability to damage the structure of fungal and bacterial cells, making it a possible alternative chemical for use in the preservation of fruits and vegetables.
RESUMEN
Oligodendrocyte precursor cell (OPC) maturation requires membrane addition for myelin sheath formation. Since the Rab system has been shown to contribute to membrane addition in other cell types, in this study, we explored the role of Rab in OPC maturation. SiRNA and shRNA techniques and conditional knockout mice provided in vitro and in vivo evidence that Rab10 is involved in OPC maturation and may affect myelination during OPC development.
Asunto(s)
Células Precursoras de Oligodendrocitos/patología , Oligodendroglía/fisiología , Proteínas de Unión al GTP rab/deficiencia , Animales , Animales Recién Nacidos , Células Cultivadas , Ratones , Ratones Noqueados , Ratones Transgénicos , Ratas , Ratas Sprague-Dawley , Proteínas de Unión al GTP rab/genéticaRESUMEN
Oligodendrocyte precursor cell (OPC) development into myelinated oligodendrocytes demands vigorous membrane addition. Since myristoylated alanine-rich C-kinase substrate (MARCKS) reportedly contributes to Ras-associated protein (Rab)-10-associated vesicle insertion into neuronal membranes, we investigated the role of MARCKS in OPC maturation. We found that either knockdown of MARCKS or interruption of its interaction with Rab10 would cause a decrease of the cell membrane area during OPC development. Enhanced MARCKS phosphorylation by Nogo66 or myelin debris treatment inhibited OPC maturation, while its dephosphorylation by protein phosphatase 2 A activator D-erythro-sphingosine promoted OPC development in the presence of myelin debris. Our results demonstrated that MARCKS is involved in OPC maturation by interacting with Rab10.
Asunto(s)
Diferenciación Celular/fisiología , Vaina de Mielina/metabolismo , Sustrato de la Proteína Quinasa C Rico en Alanina Miristoilada/metabolismo , Células Precursoras de Oligodendrocitos/citología , Oligodendroglía/citología , Animales , Membrana Celular/metabolismo , Células Cultivadas , Proteínas de la Membrana/metabolismo , Proteínas de Microfilamentos/metabolismo , Neuronas/metabolismo , Oligodendroglía/metabolismo , Ratas Sprague-DawleyRESUMEN
A series of N-(aminosulfonyl)-4-podophyllotoxin carbamates were synthesized via the Burgess-type intermediate, and their antiproliferative activities were evaluated. Most of them possessed more potent cytotoxic effects against four human tumor cell lines (HeLa, A-549, HCT-8 and HepG2) and less toxic to normal human fetal lung fibroblast WI-38 cells than etoposide. In particular, N-(morpholinosulfonyl)-4-podophyllotoxin carbamate (9) exhibited the most potent activity towards these four tumor cells with IC50 values in the range of 0.5-16.5µM. Furthermore, immunofluorescence analysis revealed that 9 induced cell apoptosis by up-regulating the expression of p53 and ROS. Meanwhile, 9 effectively inhibited tubulin polymerization and microtubule assembly at cellular levels in HeLa cells. In addition, 9 could induce cell cycle arrest in the G2/M phase in HeLa cells by up-regulating levels of cyclinB1 and cdc2 and decreasing the expression of p-cdc2. These results indicated that 9 had potential for further development as anticancer agents.
Asunto(s)
Antineoplásicos/farmacología , Carbamatos/farmacología , Podofilotoxina/análogos & derivados , Antineoplásicos/síntesis química , Antineoplásicos/química , Carbamatos/síntesis química , Carbamatos/química , Línea Celular , Proliferación Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Ensayos de Selección de Medicamentos Antitumorales , Humanos , Estructura Molecular , Podofilotoxina/síntesis química , Podofilotoxina/química , Podofilotoxina/farmacología , Relación Estructura-ActividadRESUMEN
BACKGROUND: Cervical cancer (CC) is a leading cause of mortality in females, especially in developing countries. The two viral oncoproteins E6 and E7 mediate the oncogenic activities of high-risk human papillomavirus (hrHPV), and hrHPV, especially HPV16 or/and HPV18 (HPV16/18) play critical roles in CC through different pathways. STK31 gene of which the expression has been proven to be regulated by the methylation status of its promoter, is one of the novel cancer/testis (CT) genes and plays important roles in human cancers. Reasearches have indicated that viral infection is correlated to the methylation statuses of some genes. Herein, we detected methylation status of the STK31 gene in cervical tumors and explored its interaction with HPV16 or/and HPV18 (HPV16/18) infection. METHODS: Bisulfite genomic sequencing PCR (BGS) combined with TA clone, methylation-specific PCR (MSP) were used to analyze methylation statuses of the STK31 gene promoter/exon 1 region in HPV16/18-positive, HPV-negative CC cell lines; ectopically expressed HPV16 E6, -E7, and -E6/E7 CC cells; normal cervical tissues and cervical tumor tissues of different stages. The mRNA and protein expressions of STK31 were detected by RT-PCR and western blotting. RESULTS: The STK31 gene promoter/exon 1 was hypomethylated in the HPV16/18-positive cell lines HeLa, SiHa and CaSki, and the mRNA and protein expression were detected. In contrast, the STK31 gene exhibited hypermethylation and silenced expression in the HPV-negative CC cells C33A and HT-3. Compared with the primary HPV-negative CC cell lines, the STK31 methylation was downregulated, and STK31 expression was induced in the HPV16E7/E67 transfected cells. The methylation statuses and expressions of STK31 were verified in the cervical tumor samples at different stages. Additionally, chemotherapy treatment may influence STK31 expression by regulating its methylation status. CONCLUSIONS: STK31 may be a novel cellular target gene for the HPV16 oncogeneE7. The HPV16 oncogene E7 may affect STK31 expression through a methylation-mediated mechanism. The aberrant methylation of the STK31 promoter/exon 1 region may be a precursor of human cervical carcinogenesis and a potential DNA aberrant methylation biomarker of conditions ranging from precancerous disease to invasive cancer.
Asunto(s)
Metilación de ADN , Interacciones Huésped-Patógeno , Papillomavirus Humano 16/fisiología , Proteínas E7 de Papillomavirus/metabolismo , Regiones Promotoras Genéticas , Proteínas Serina-Treonina Quinasas/biosíntesis , Neoplasias del Cuello Uterino/patología , Adulto , Biomarcadores de Tumor/análisis , Western Blotting , Línea Celular Tumoral , ADN/química , Proteínas de Unión al ADN/metabolismo , Femenino , Perfilación de la Expresión Génica , Papillomavirus Humano 18/fisiología , Humanos , Persona de Mediana Edad , Proteínas Oncogénicas Virales/metabolismo , Reacción en Cadena en Tiempo Real de la Polimerasa , Neoplasias del Cuello Uterino/virologíaRESUMEN
A series of deoxypodophyllotoxin-5-fluorouracil hybrid compounds were synthesized, and their cytotoxic activity was evaluated using four human cancer cell lines (HeLa, A549, HCT-8, and HepG2) and the human normal cell line WI-38. The synthesized compounds exhibited greater cytotoxic activity in tumor cells and reduced toxicity in the normal cell line compared with the anticancer drug VP-16 and 5-FU. Additionally, the most potent of these compounds-4'-O-demethyl-4-deoxypodophyllotoxin-4'-yl 4-((6-(2-(5-fluorouracil-yl) acetamido) hexyl) amino)-4-oxobutanoate (compound 22)-induced cell-cycle arrest in the G2/M phase by regulating levels of cdc2, cyclinB1, and p-cdc2 in A549 cells. Furthermore, compound 22 may inhibited the migration of A549 cells via down-regulation of MMP-9 and up-regulation of TIMP-1.
Asunto(s)
Antineoplásicos/síntesis química , Antineoplásicos/farmacología , Puntos de Control del Ciclo Celular/efectos de los fármacos , Movimiento Celular/efectos de los fármacos , Podofilotoxina/análogos & derivados , Antineoplásicos/química , Línea Celular , Proliferación Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Ensayos de Selección de Medicamentos Antitumorales , Humanos , Metaloproteinasa 9 de la Matriz/metabolismo , Estructura Molecular , Podofilotoxina/síntesis química , Podofilotoxina/química , Podofilotoxina/farmacología , Relación Estructura-Actividad , Inhibidor Tisular de Metaloproteinasa-1/metabolismoRESUMEN
LINGO-1 is a transmembrane receptor expressed primarily in the central nervous system (CNS) and plays an important role in myelination. Recent studies have indicated that it is also involved in oligodendrocyte precursor cell (OPC) survival and differentiation; however, the downstream signaling pathway underlying OPC development is unknown. In our previous study, we found that LINGO-1 is associated with WNK1 in mediating Nogo-induced neurite extension inhibition by RhoA activation. In an effort to identify the role of LINGO-1-WNK1 in OPCs, we first confirmed that WNK1 is also expressed in OPCs and co-localized with LINGO-1, which suppresses WNK1 expression by RNA interference-attenuated Nogo66-induced inhibition of OPC differentiation. Furthermore, we mapped the WNK1 kinase domain using several fragmented peptides to identify the key region of interaction with LINGO-1. We found that a sequence corresponding to the D6 peptide is necessary for the interaction. Finally, we found that using the TAT-D6 peptide to introduce D6 peptide into primary cultured OPC inhibits the association between LINGO-1 and WNK1 and significantly attenuates Nogo66-induced inhibition of OPC differentiation. Taken together, our results show that WNK1, via a specific region on WNK1 kinase domain, interacts with LINGO-1, thus mediating Nogo66-inhibited OPC differentiation.
Asunto(s)
Proteínas de la Mielina/metabolismo , Células-Madre Neurales/metabolismo , Neurogénesis , Oligodendroglía/metabolismo , Proteínas Serina-Treonina Quinasas/metabolismo , Animales , Sitios de Unión , Células Cultivadas , Proteínas de la Membrana/metabolismo , Antígenos de Histocompatibilidad Menor , Proteínas de la Mielina/genética , Proteínas del Tejido Nervioso/metabolismo , Células-Madre Neurales/citología , Proteínas Nogo , Oligodendroglía/citología , Unión Proteica , Proteínas Serina-Treonina Quinasas/química , Proteínas Serina-Treonina Quinasas/genética , Ratas , Ratas Sprague-Dawley , Proteína Quinasa Deficiente en Lisina WNK 1RESUMEN
OBJECTIVE: To investigate the efficacy of volume-targeted ventilation (VTV) for the treatment of neonatal respiratory distress syndrome (NRDS). METHODS: Fifty-two neonates with NRDS between August 2013 and August 2015 were randomly divided into two groups: VTV and pressure-controlled ventilation (PCV) (n=26 each ). A/C+Vc+ ventilation model was applied in the VTV group, and A/C+PCV ventilation model was applied in the PCV group. Arterial blood gas analysis was performed at 6, 24, and 48 hours after ventilation. The following parameters were observed: time of invasive ventilation, duration of oxygen therapy, mortality, and the incidence rates of hypocapnia, pneumothorax, ventilator-associated pneumonia (VAP), grade III-IV periventricular-intraventricular hemorrhage (PVH-IVH), periventricular leukomalacia (PVL), bronchopulmonary dysplasia (BPD), and retinopathy of prematurity (ROP). RESULTS: Compared with the PCV group, the VTV group had a significantly shorter time of invasive ventilation (P<0.05) and significantly lower incidence rates of hypocapnia, VAP, and PVL (P<0.05); however, there were no significant differences in the duration of oxygen therapy, mortality, and incidence rates of pneumothorax, grade III-IV PVH-IVH, BPD, and ROP. CONCLUSIONS: VTV has a better efficacy than PCV in the treatment of NRDS, and is worthy of clinical promotion and application.
Asunto(s)
Respiración Artificial/métodos , Síndrome de Dificultad Respiratoria del Recién Nacido/terapia , Femenino , Humanos , Recién Nacido , Masculino , Respiración Artificial/efectos adversosRESUMEN
Axon development and elongation require strictly controlled new membrane addition. Previously, we have shown the involvement of Rab10 in directional membrane insertion of plasmalemmal precursor vesicles (PPVs) during neuronal polarization and axonal growth. However, the mechanism responsible for PPV transportation remains unclear. Here we show that c-Jun N-terminal kinase-interacting protein 1 (JIP1) interacts with GTP-locked active form of Rab10 and directly connects Rab10 to kinesin-1 light chain (KLC). The kinesin-1/JIP1/Rab10 complex is required for anterograde transport of PPVs during axonal growth. Downregulation of JIP1 or KLC or disrupting the formation of this complex reduces anterograde transport of PPVs in developing axons and causes neuronal polarity defect. Furthermore, this complex plays an important role in neocortical neuronal polarization of rats in vivo. Thus, this study has demonstrated a mechanism underlying directional membrane trafficking involved in axon development.
Asunto(s)
Proteínas Adaptadoras Transductoras de Señales/metabolismo , Polaridad Celular/fisiología , Neuronas/fisiología , Proteínas de Unión al GTP rab/metabolismo , Proteínas Adaptadoras Transductoras de Señales/genética , Animales , Axones/metabolismo , Encéfalo/citología , Células Cultivadas , Estructuras Celulares/metabolismo , Embrión de Mamíferos , Femenino , Regulación de la Expresión Génica/fisiología , Humanos , Antígeno Ki-67/metabolismo , Cinesinas , Proteínas Luminiscentes/genética , Proteínas Luminiscentes/metabolismo , Masculino , Microscopía Confocal , Proteínas Asociadas a Microtúbulos/genética , Proteínas Asociadas a Microtúbulos/metabolismo , Neuronas/citología , Transporte de Proteínas/fisiología , ARN Interferente Pequeño/genética , ARN Interferente Pequeño/metabolismo , Ratas , Ratas Sprague-Dawley , Proteínas de Dominio T Box/metabolismo , Proteínas de Unión al GTP rab/genéticaRESUMEN
Mitogen-activated protein kinases (MAPKs) play important roles in stress responses and development in plants. Maize (Zea mays), an important cereal crop, is a model plant species for molecular studies. In the last decade, several MAPKs have been identified in maize; however, their functions have not been studied extensively. Genome-wide identification and expression analysis of maize MAPK genes could provide valuable information for understanding their functions. In this study, 20 non-redundant maize MAPK genes (ZmMPKs) were identified via a genome-wide survey. Phylogenetic analysis of MAPKs from maize, rice (Oryza sativa), Arabidopsis (Arabidopsis thaliana), poplar (Populus trichocarpa), and tomato (Solanum lycopersicum) classified them into four major classes. ZmMPKs in the same class had similar domains, motifs, and genomic structures. Gene duplication investigations suggested that segmental duplications made a large contribution to the expansion of ZmMPKs. A number of cis-acting elements related to plant development and response to stress and hormones were identified in the promoter regions of ZmMPKs. Furthermore, transcript profile analysis in eight tissues and organs at various developmental stages demonstrated that most ZmMPKs were preferentially expressed in reproductive tissues and organs. The transcript abundance of most ZmMPKs changed significantly under salt, drought, cold, or abscisic acid (ABA) treatments, implying that they might participate in abiotic stress and ABA signaling. These expression analyses indicated that ZmMPKs might serve as linkers between abiotic stress signaling and plant reproduction. Our data will deepen our understanding of the complexity of the maize MAPK gene family and provide new clues to investigate their functions.
Asunto(s)
Regulación Enzimológica de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Proteínas Quinasas Activadas por Mitógenos/genética , Transducción de Señal/genética , Estrés Fisiológico/genética , Zea mays/enzimología , Zea mays/genética , Ácido Abscísico/farmacología , Secuencias de Aminoácidos , Cromosomas de las Plantas/genética , Frío , Secuencia Conservada/genética , Sequías , Exones/genética , Duplicación de Gen , Perfilación de la Expresión Génica , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Genes de Plantas , Intrones/genética , Proteínas Quinasas Activadas por Mitógenos/química , Proteínas Quinasas Activadas por Mitógenos/metabolismo , Familia de Multigenes , Especificidad de Órganos/efectos de los fármacos , Especificidad de Órganos/genética , Filogenia , Proteínas de Plantas/química , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Regiones Promotoras Genéticas/genética , Estructura Terciaria de Proteína , Reproducción/efectos de los fármacos , Reproducción/genética , Cloruro de Sodio/farmacología , Estrés Fisiológico/efectos de los fármacos , Zea mays/efectos de los fármacosRESUMEN
Gonadotropin-releasing hormone (GnRH) neurons play a pivotal role in reproductive function. GnRH is released in distinct pulses that are regulated by neurotransmitters or neuromodulators. With immunohistochemistry and GAD67-GFP knockin mice, this study shows for the first time that a subset of GnRH neurons in the forebrain of adult mouse is γ-aminobutyric acid (GABA)-ergic. There is a gender difference in the percentage of GnRH neurons expressing GAD67-GFP in female vs. male mice. The percentage of GnRH neurons expressing GAD67-GFP decreased after castration of female mice and increased to the normal female level after estradiol treatment. The percentage of GnRH neurons expressing GAD67-GFP did not change significantly in intact, castrated, or castration + testosterone propionate-treated male mice. During the female estrous cycle, the percentage of GnRH neurons expressing GAD67-GFP was higher during the estrous stage than during the diestrous stage. During sexual maturation of postnatal development, GnRH neurons did not express GAD67-GFP until postnatal day (P) 15, and the gender differences were first detected at P30, which corresponds to the maturation stage. In conclusion, our data suggest that 1) a subset of GnRH neurons in mouse forebrain is GABA-ergic, 2) expression of GAD67-GFP in GnRH neurons is at least in part regulated by estrogen, and 3) GnRH neurons secrete GABA to regulate themselves.
Asunto(s)
Hormona Liberadora de Gonadotropina/metabolismo , Neuronas/metabolismo , Prosencéfalo/citología , Ácido gamma-Aminobutírico/metabolismo , Factores de Edad , Animales , Animales Recién Nacidos , Castración , Estradiol/farmacología , Femenino , Regulación del Desarrollo de la Expresión Génica/efectos de los fármacos , Regulación del Desarrollo de la Expresión Génica/genética , Glutamato Descarboxilasa/genética , Glutamato Descarboxilasa/metabolismo , Proteínas Fluorescentes Verdes/genética , Proteínas Fluorescentes Verdes/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Neuronas/citología , Neuronas/efectos de los fármacos , Propionato de Testosterona/farmacologíaRESUMEN
Peripheral nerve injury (PNI) is a common disease, which results in a partial or total loss of motor, sensory and autonomic functions, leading to a decrease in quality of life. Schwann cells play a vital role in maintaining the peripheral nervous system and in injury and repair. Using immunohistochemistry, Western blot, calcium assay and bromodeoxyuridine (BrdU) proliferation assay, the present study clearly demonstrated that P2X7 receptors (R) were expressed in myelinating and non-myelinating Schwann cells in longitudinal sections of sciatic nerves. After sciatic nerve injury (SNI), P2X7R expression in Schwann cells of injured sciatic nerves was significantly up-regulated during the early days of SNI. Double immunofluorescence of proliferating cell nuclear antigen (PCNA) and P2X7R implied that P2X7R may be involved in proliferation of Schwann cells. Further experiments on primary cultures of Schwann cells showed that P2X7R are functionally expressed in Schwann cells of rat sciatic nerves; ATP via P2X7R can promote Schwann cell proliferation, possibly via the MAPK/ERK intracellular signalling pathway. Other possible roles of P2X7R on Schwann cells are discussed.
Asunto(s)
Proliferación Celular , Ganglios Espinales/metabolismo , Regeneración Nerviosa/fisiología , Receptores Purinérgicos P2X7/metabolismo , Células de Schwann/metabolismo , Nervio Ciático/lesiones , Animales , Proliferación Celular/fisiología , Masculino , Ratas Sprague-Dawley , Células de Schwann/citología , Neuropatía Ciática/patología , Regulación hacia Arriba/fisiologíaRESUMEN
Transient receptor potential melastatin-7 (TRPM7) channels are involved in many cellular physiological and pathological processes. The present study was designed to investigate the expression of TRPM7 channels and the potential role in regulating cell proliferation and adipogenesis in 3T3-L1 preadipocytes with approaches of whole-cell patch voltage-clamp, molecular biology, cell proliferation, adipogenesis, etc. We found that a TRPM7-like current was recorded with Mg(2+) -free pipette solution in 3T3-L1 preadipocytes, and the current was inhibited by intercellular free Mg(2+) . The TRPM7-like current was potentiated by acidic pH and inhibited by 2-aminoethoxydiphenyl borate (2-APB). RT-PCR, Western blot and immunocytochemistry revealed that gene and protein of TRPM7 channels were abundant in 3T3-L1 preadipocytes. Blockade of TRPM7 channels with 2-APB inhibited cell proliferation in 3T3-L1 cells. In addition, knockdown of TRPM7 with specific siRNA inhibited both proliferation and adipogenesis. The present study demonstrates for the first time that TRPM7 channels regulate cell cycle and adipogenesis of 3T3-L1 preadipocytes.
Asunto(s)
Adipogénesis/genética , Canales Catiónicos TRPM/genética , Células 3T3-L1 , Animales , Compuestos de Boro/farmacología , Proliferación Celular/efectos de los fármacos , Técnicas de Silenciamiento del Gen , Ratones , Técnicas de Placa-Clamp , Canales Catiónicos TRPM/antagonistas & inhibidores , Canales Catiónicos TRPM/biosíntesisRESUMEN
OBJECTIVE: To observe the morphological change in the pathological process of Echinococcus multilocularis in liver of infected rats after treatment with liposome entrapped albendazole and iodized oil suspension through portal vein, and investigate its efficacy against hepatic alveolar echinococcosis. METHODS: One Echinococcus multilocularis-infected rat served as control. Another 19 infected rats were infused with 0.2ml liposome entrapped albendazole and iodized oil suspension through portal vein. At 4, 7 and 10 d after treatment, the rats were sacrificed for further observation. The E. multilocularis metacestodes were collected, and the pathological change of alveolar hydatid was observed with hematoxylin-eosin staining and toluidine blue staining. RESULTS: In four days post treatment, E. multilocularis metacestodes were normal On the 7th day and 10th day post treatment, the major pathological change was degeneration and necrosis, respectively. A great quantity of liposomes and iodized oil deposited in the metacestodes, and caused breakdown of E. multilocularis tissue, collapse of cysts, cataplasia of germinal layer and laminated layer. CONCLUSION: Interventional therapy trans-hepatic vein may be an effective way to treat hepatic alveolar echinococcosis.
Asunto(s)
Equinococosis Hepática/patología , Echinococcus multilocularis , Vena Porta/patología , Albendazol , Animales , Equinococosis Hepática/terapia , Embolización Terapéutica , Necrosis , RatasRESUMEN
BACKGROUND: Tissue invasion and metastasis are acquired abilities of cancer and related to the death in oral squamous cell carcinoma (OSCC). Emerging observations indicate that the epithelial-to-mesenchymal transition (EMT) is associated with tumor progression and the generation of cells with cancer stem cells (CSCs) properties. Membrane Type 1 Matrix Metalloproteinase (MT1-MMP) is a cell surface proteinase, which is involved in degrading extracellular matrix components that can promote tumor invasion and cell migration. METHODS: In the current study, we utilized SCC9 cells stably transfected with an empty vector (SCC9-N) or a vector encoding human MT1-MMP (SCC9-M) to study the role of MT1-MMP in EMT development. RESULTS: Upon up-regulation of MT1-MMP, SCC9-M cells underwent EMT, in which they presented a fibroblast-like phenotype and had a decreased expression of epithelial markers (E-cadherin, cytokeratin18 and ß-catenin) and an increased expression of mesenchymal markers (vimentin and fibronectin). We further demonstrated that MT1-MMP-induced morphologic changes increased the level of Twist and ZEB, and were dependent on repressing the transcription of E-cadherin. These activities resulted in low adhesive, high invasive abilities of the SCC9-M cells. Furthermore, MT1-MMP-induced transformed cells exhibited cancer stem cell (CSC)-like characteristics, such as low proliferation, self-renewal ability, resistance to chemotherapeutic drugs and apoptosis, and expression of CSCs surface markers. CONCLUSIONS: In conclusion, our study indicates that overexpression of MT1-MMP induces EMT and results in the acquisition of CSC-like properties in SCC9 cells. Our growing understanding of the mechanism regulating EMT may provide new targets against invasion and metastasis in OSCC.