RESUMEN
Objective: To explore the method and effect of endoscopic assisted functional rhinoplasty for patients with deviated nose and deviated nasal septum, which achieve correction of nasal morphology and ventilation dysfunction. Methods: The clinical data of 226 patients with deviated nose and deviated nasal septum from June 2009 to February 2022 who were treated by endoscopic assisted functional rhinoplasty in the Affiliated Hospital of Qingdao University were analyzed retrospectively. There were 174 males and 52 females, with the age ranging from 7 to 67 years old. The effect was evaluated by subjective and objective evaluation methods. SPSS 27.0 software was used for statistical analysis. Results: All patients were followed up for 6 to 24 months, 174 cases were cured (174/226, 76.99%), 52 cases were effective (52/226, 23.01%), and the total effective rate was 100% (226/226). The difference between preoperative and postoperative facial appearance deviation was statistically significant ((6.84±2.25)mm vs (1.82±1.05)mm, t=38.94, P<0.001), and the nasal ventilation function of all patients was improved. Conclusions: Endoscopic assisted functional rhinoplasty for the patients with deviated nose combined with deviated nasal septum has the advantages of clear surgical field, fewer complications, and good result. It can achieve the purpose of simultaneous correction of nasal and ventilation dysfunction, which is recommended for popularizing in clinical application.
RESUMEN
The proliferation of vascular smooth muscle cells plays a crucial role in pathogenesis of cardiovascular disease. The principal objective of this study was to determine the effects of Ojeoksan (OJS) on human aortic smooth muscle cell (HASMC) proliferation induced by tumor necrosis factor α (TNF-aα). Thymidine incorporation after TNF-α treatment was increased and this effect was inhibited significantly by OJS treatment. HASMC proliferation and migration by kinetic live cell imaging were also reduced by treatment with OJS. TNF-α induced the expression of cyclins/cyclin-dependent kinases (CDKs) and reduced the expression of p21waf1/cip1/p27kip1. However, OJS also attenuated the expression of TNF-α-induced cell-cycle regulatory proteins. The results of Western blot analysis demonstrated that the TNF-α treated HASMC secreted gelatinases, probably including MMP-2/-9, which may be involved in the invasion and migration of HASMC. Additionally, OJS suppressed the mRNA expression levels of matrix metalloproteinase-2/-9 (MMP-2/-9) in a dose-dependent manner. OJS inhibited the production of TNF-α-induced hydrogen peroxide (H2O2) and the formation of DCF-sensitive intracellular reactive oxygen species (ROS). Further, OJS suppressed the nuclear translocation and phosphorylation of inhibitor of kappa B-α (IκB-α) of nuclear factor κB (NF-κB) under TNF-α conditions. Our results demonstrate that OJS exerts inhibitory effects on TNF-α-induced HASMC proliferation and migration, suggesting the involvement of the inhibition of both MMP-2 and MMP-9 expressions, and the downregulation of ROS/NF-κB signaling. Thus, herbal decoction OJS may be a possible therapeutic approach to the inhibition of cardiovascular disease including atherosclerosis.
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Movimiento Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Medicamentos Herbarios Chinos/farmacología , Músculo Liso Vascular/efectos de los fármacos , Miocitos del Músculo Liso/efectos de los fármacos , Aorta/efectos de los fármacos , Aorta/metabolismo , Células Cultivadas , Inhibidor p27 de las Quinasas Dependientes de la Ciclina/metabolismo , Regulación hacia Abajo/efectos de los fármacos , Humanos , Peróxido de Hidrógeno/farmacología , Metaloproteinasa 2 de la Matriz/metabolismo , Metaloproteinasa 9 de la Matriz/metabolismo , Músculo Liso Vascular/metabolismo , Miocitos del Músculo Liso/metabolismo , FN-kappa B/metabolismo , Fosforilación/efectos de los fármacos , Especies Reactivas de Oxígeno/metabolismo , Transducción de Señal/efectos de los fármacos , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
Euphorbia humifusa Willd. (EH), rich in flavonoids, has long been used for the treatment of bacillary dysentery and enteritis in China, and is known to have antioxidant, hypotensive and hypolipidemic properties. However, the vasorelaxant effect of total flavonoids of EH (TFEH) and action mechanisms are not clearly defined yet. The aim of the present study was to investigate the effects of TFEH on the vascular tension and its underlying mechanisms. Experiments were performed in rat thoracic aorta using the organ bath system. TFEH (0.01 - 100 µg/ml) caused a concentration-dependent vasorelaxation, which was dependent on a functional endothelium, and were significantly attenuated by inhibitors of endothelial NO synthase, its upstream signaling pathway, PI3K/Akt, and soluble guanylate cyclase, but not by blockade of KCa channel, KATP channel, cyclooxygenase, muscarinic and ß-adrenergic receptors. Extracellular Ca2+ depletion, and pre-treatment with modulators of the store-operated Ca2+ entry channels, Gd3+ and 2-aminoethyl diphenylborinate, significantly attenuated the TFEH-induced vasorelaxation. Our findings suggest that TFEH elicit vasorelaxation via endothelium-dependent NO-cGMP pathway through activation of PI3K/Akt- and Ca2+-eNOS-NO signaling. Further, it is suggested that TFEH-induced activation of the NO-soluble guanylate cyclase-cGMP-protein kinase G signaling relaxes vascular smooth muscle cells through an inhibition of the L-type Ca2+ channel activity.
Asunto(s)
Aorta Torácica/efectos de los fármacos , Euphorbia/química , Flavonoides/farmacología , Vasodilatación/efectos de los fármacos , Vasodilatadores/farmacología , Animales , Aorta Torácica/metabolismo , Calcio/metabolismo , Canales de Calcio/metabolismo , GMP Cíclico/metabolismo , Medicamentos Herbarios Chinos/farmacología , Endotelio Vascular/efectos de los fármacos , Endotelio Vascular/metabolismo , Óxido Nítrico/metabolismo , Óxido Nítrico Sintasa de Tipo III/metabolismo , Fosfatidilinositol 3-Quinasas/metabolismo , Extractos Vegetales/farmacología , Prostaglandina-Endoperóxido Sintasas/metabolismo , Ratas , Ratas Sprague-Dawley , Transducción de Señal/efectos de los fármacos , Guanilil Ciclasa Soluble/metabolismoRESUMEN
Objective: To analyze the clinical features and the pathogenetic law of traumatic optic neuropathy through epidemiologic study. Methods: 265 cases (275 eyes) with TON treated in Department of Otorhinolaryngology Head and Neck Surgery, the Affiliated Hospital of Qingdao University from April 1999 to August 2015 were retrospectively analyzed. Multiple Logistic regression analyses were used to evaluate potential prognostic factors on visual acuity. Results: TON occured mostly in young (194/265, 73.21%) man (235/265, 88.68%), the majority of patients came from villages and towns (209/265, 78.87%). Traffic accident (197/265, 74.34%) remained the main etiology, with strike (36/265, 13.58%) and fall (17/265, 6.42%) as the common etiology. Most patients had head injuries. The effective rate of vision improvement was 53.45%(147/275). Multiple logistic regression analyses identified that initial visual acuity with light perception or better vision, optic canal fracture and orbital wall fracture were visual acuity key factors of TON (χ(2) value was 24.674, 19.755, 9.175, respectively, all P<0.01), initial visual acuity with light perception or better vision was the protective factor on visual acuity recovery (OR=5.008, P<0.001), the presence of optic canal fracture and orbital wall fracture were the risk factors on visual acuity recovery (OR value was 0.110, 0.329, respectively, all P<0.01). Conclusions: Ton occurs mostly in young man because of traffic accident. Visual impairment of TON is severe. The suitable preventive measures should be carried out according to its epidemiological features.
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Traumatismos del Nervio Óptico/etiología , Accidentes por Caídas/estadística & datos numéricos , Accidentes de Tránsito/estadística & datos numéricos , Factores de Edad , Traumatismos Craneocerebrales/complicaciones , Descompresión Quirúrgica , Femenino , Humanos , Masculino , Fracturas Orbitales/complicaciones , Análisis de Regresión , Estudios Retrospectivos , Factores de Riesgo , Factores Sexuales , Agudeza VisualRESUMEN
Rumex acetosa L. (RA) (Polygonaceae) is an important traditional Chinese medicine (TCM) commonly used in clinic for a long history in China and the aerial parts of RA has a wide variety of pharmacological actions such as diuretic, anti-hypertensive, anti-oxidative, and anti-cancer effects. However, the mechanisms involved are to be defined. The purpose of the present study was to evaluate the vasorelaxant effect and define the mechanism of action of the ethanol extract of Rumex acetosa L. (ERA) in rat aorta. ERA was examined for its vascular relaxant effect in isolated phenylephrine-precontracted rat thoracic aorta and its acute effects on arterial blood pressure. In addition, the roles of the nitric oxide synthase (NOS)-nitric oxide (NO) signaling in the ERA-induced effects were tested in human umbilical vein endothelial cells (HUVECs). The phosphorylation levels of Akt and eNOS were assessed by Western blot analysis in the cultured HUVECs. ERA induced endothelium-dependent vasorelaxation. The ERA-induced vasorelaxation was abolished by L-NAME (an NOS inhibitor) or ODQ (a sGC inhibitor), but not by indomethacin. Inhibition of PI3-kinase/Akt signaling pathway markedly reduced the ERA-induced vasorelaxation. In HUVECs, ERA increased NO formation in a dose-dependent manner, which was inhibited by L-NAME and by removing extracellular Ca(2+). In addition, ERA promoted phosphorylation of Akt and eNOS, which was prevented by wortmannin and LY294002, indicating that ERA induces eNOS phosphorylation through the PI3-kinase/Akt pathway. Further, in anesthetized rats, intravenously administered ERA decreased arterial blood pressure in a dose-dependent manner through an activation of the NOS-NO system. In summary, the ERA- induced vasorelaxation was dependent on endothelial integrity and NO production, and was mediated by activation of both the endothelial PI3-kinase/Akt- and Ca(2+)-eNOS-NO signaling and muscular NO-sGC-cGMP signaling.
Asunto(s)
Aorta/efectos de los fármacos , Células Endoteliales de la Vena Umbilical Humana/efectos de los fármacos , Extractos Vegetales/farmacología , Rumex , Vasodilatadores/farmacología , Animales , Aorta/fisiología , Calcio/metabolismo , Células Cultivadas , GMP Cíclico/metabolismo , Células Endoteliales de la Vena Umbilical Humana/metabolismo , Humanos , Técnicas In Vitro , Masculino , Medicina Tradicional China , Óxido Nítrico/metabolismo , Óxido Nítrico Sintasa de Tipo III/metabolismo , Fosfatidilinositol 3-Quinasa/metabolismo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Ratas Sprague-Dawley , Transducción de Señal/efectos de los fármacos , Vasodilatación/efectos de los fármacosRESUMEN
Rubus chingii Hu (Rosaceae) is an important traditional Chinese medicine that has been used to improve function of the kidney and treat excessive polyuria. However, the effects of Rubus chingii on the cardiovascular system and its pharmacological mechanisms of action have not been studied. The aim of the present study was to evaluate the cardiovascular effects of ethanol extract of Rubus chingii (ERC) in rats. The changes in systolic blood pressure and heart rate of rats and vascular tone of aortic rings in in vitro were measured using pressure transducer and force transducer, respectively, connected to a multichannel recording system. ERC decreased systolic blood pressure and heart rate in a concentration-dependent manner. ERC induced vasorelaxation in a concentration-dependent manner. The ERC-induced vasorelaxation was not observed in the absence of the endothelium. The vasorelaxant effect of ERC was significantly attenuated by inhibition of endothelial NO synthase (eNOS), soluble guanylyl cyclase (sGC), or Ca(2+) entry from extracellular sources with L-NAME, ODQ, diltiazem, or extracellular Ca(2+) depletion, respectively. Similarly, an inhibition of Akt with wortmannin attenuated the ERC-induced vasorelaxation. Modulators of the store-operated Ca(2+) entry, thapsigargin, Gd(3+), and 2-aminoethyl diphenylborinate markedly attenuated the ERC-induced vasorelaxation. Furthermore, 4-aminopyridine an inhibitor of voltage-dependent K(+) (KV) channel, significantly attenuated the ERC-induced vasorelaxation. However, tetraethylammonium and glibenclamide, had no significant effect on the ERC-induced vasorelaxation. Indomethacin, atropine, and propranolol had no effects on the ERC-induced vasorelaxation. The present study demonstrates that ERC induces vasorelaxation via endothelium-dependent two-step signaling: an activation of the Ca(2+)-eNOS-NO signaling in the endothelial cells and then subsequent stimulation of the NO-sGC-cGMP-KV channel signaling in the vascular smooth muscle cells. The Akt-eNOS pathway is also suggested to be involved in this relaxation. Also, the findings suggest that the ERC-induced vasorelaxation is closely related to the hypotensive action of the agent.
Asunto(s)
Sistema Cardiovascular/efectos de los fármacos , Medicamentos Herbarios Chinos/farmacología , Etanol/química , Frutas/química , Extractos Vegetales/química , Extractos Vegetales/farmacología , Rosaceae/química , Animales , Aorta Torácica/efectos de los fármacos , Presión Sanguínea/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Medicamentos Herbarios Chinos/química , Medicamentos Herbarios Chinos/aislamiento & purificación , Frecuencia Cardíaca/efectos de los fármacos , Masculino , Mesotelina , Extractos Vegetales/aislamiento & purificación , Ratas , Ratas Sprague-Dawley , Vasodilatación/efectos de los fármacosRESUMEN
OBJECTIVE: Increasing evidence indicates that MicroRNAs, a class of small RNA molecules, play crucial roles in tumorigenesis, through affecting cell proliferation, apoptosis and metastasis. The present study aimed to investigate the effect of miR-205 on gastric cancer cell proliferation. MATERIALS AND METHODS: The expression of miR-205 was examined in the gastric cancer tissues and cell lines. BrdU incorporation assay was used to measure the cell proliferation. Western blot was performed to determine the protein expression. RESULTS: miR-205 is significantly down-regulated in gastric cancer tissues, compared with adjacent normal tissues. Besides, miR-205 expression is associated with clinical and pathological characteristics of patients. In vitro studies further found that inhibition of miR-205 significantly promoted gastric cancer cell proliferation via cell-cycle progression. Further analyses indicated that miR-205 was able to repress oncoprotein Yin Yang 1 expression, through targeting its 3'-untranlated region. CONCLUSIONS: Our data suggest that down-regulation of miR-205 may represent an important mechanism for the development of gastric cancer.
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MicroARNs/genética , Neoplasias Gástricas/genética , Línea Celular Tumoral , Proliferación Celular , Regulación hacia Abajo , Mucosa Gástrica/metabolismo , Humanos , Masculino , MicroARNs/metabolismo , Persona de Mediana Edad , Análisis de Secuencia por Matrices de Oligonucleótidos , Reacción en Cadena en Tiempo Real de la Polimerasa , Neoplasias Gástricas/metabolismo , Neoplasias Gástricas/patología , Factor de Transcripción YY1/genética , Factor de Transcripción YY1/metabolismoRESUMEN
The complete coding sequences (CDSs) of "Yunnan Purple Pepper No.1" (Capsicum annuum L.) AN2 and UPA20 genes were amplified using the reverse transcriptase polymerase chain reaction on the basis of the conserved sequence information of some Solanaceae plants and known highly homologous pepper expressed sequence tags. The nucleotide sequence analysis of these 2 genes revealed that pepper AN2 gene encoded a protein of 263 amino acids that has high homology with the AN2-like protein of 4 species: tobacco, tomato, potato, and petunia. The UPA20 gene encoded a protein of 341 amino acids that has high homology with the proteins of 3 species: tobacco, petunia, and tomato. The tissue expression analysis indicated that the pepper AN2 gene was overexpressed in the pericarp and placenta; moderately in stems, flowers, and seeds; and weakly in the roots, leaves, and pericarp. The pepper UPA20 gene was overexpressed in the flowers and seeds; moderately expressed in the roots and stems; and weakly expressed in the leaves and placenta. Our findings might form the basis for further research on these 2 pepper genes.
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Capsicum/genética , Regulación de la Expresión Génica de las Plantas , Proteínas de Plantas/biosíntesis , China , Clonación Molecular , Flores/genética , Flores/metabolismo , Hojas de la Planta/genética , Hojas de la Planta/metabolismo , Proteínas de Plantas/genética , Análisis de Secuencia de ADN , Distribución TisularRESUMEN
A higher prevalence of depression in systemic lupus erythematosus (SLE) patients has been reported, though the mechanism underlying this phenomenon remains unclear. The present study was conducted to explore whether the polymorphism and methylation status of the serotonin transporter gene (5HTT) promoter region (PR-5HTT) contribute to depression in SLE patients from both genetic and epigenetic perspectives. In this study, 96 SLE patients and 96 healthy controls (HCs) were recruited. Depression levels of all subjects were evaluated using the Hamilton Depression Rating Scale (HDRS). The serotonin transporter-linked polymorphism (5HTTLPR) and the DNA methylation status of PR-5HTT were detected in peripheral lymphocytes of SLE patients and HCs. The differences in 5HTTLPR and DNA methylation of PR-5HTT between SLEs and HCs were compared. In SLE patients, the frequencies of short allele (S) and SS genotype of 5HTTLPR were higher in depressive SLE (SLE-D) patients than in non-depressive SLE (SLE-ND) patients. The mean HDRS score of SS homozygote patients was higher than that of patients with SL/LL genotypes. Conversely, PR-5HTT was hypomethylated in HCs as well as SLE patients. There was no difference in the methylation status between HCs and SLEs. Thus, the functional expression of PR-5HTT may be primarily regulated by gene polymorphism and not by DNA methylation. The risk allele of 5HTTLPR appears to be a major contributor to depression in SLE patients.
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Metilación de ADN , Depresión/genética , Lupus Eritematoso Sistémico/genética , Polimorfismo Genético , Regiones Promotoras Genéticas , Proteínas de Transporte de Serotonina en la Membrana Plasmática/genética , Adulto , Secuencia de Bases , Islas de CpG , Quinasas MAP Reguladas por Señal Extracelular/fisiología , Femenino , Genotipo , Humanos , Masculino , Datos de Secuencia MolecularRESUMEN
We isolated two transcription factor genes, heterotrimeric G protein beta 2 subunit (Gß2) and ArcA1, from pepper (Capsicum annuum). The complete coding sequences were amplified using reversed transcriptase PCR based on conserved sequence information of Solanum lycopersicum and several other plant species. Nucleotide sequence analysis of these two genes revealed that the pepper Gß2 gene encodes a protein of 376 amino acids that belongs to the WD40 superfamily. Tissue expression analysis indicated that this gene is highly expressed in the pericarp, moderately expressed in stem, flower, placenta, and leaves, and weakly expressed in seed. There was no expression in the roots. The ArcA1 gene encodes a protein of 331 amino acids that also belongs to the WD40 superfamily. Tissue expression analysis indicated that the pepper ArcA1 gene is moderately expressed in the pericarp and weakly expressed in seed. There was no expression in root, stem, flower, placenta, or leaves.
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Capsicum/genética , Subunidades beta de la Proteína de Unión al GTP/genética , Proteínas de Plantas/genética , Secuencia de Aminoácidos , Secuencia de Bases , Capsicum/metabolismo , Clonación Molecular , Subunidades beta de la Proteína de Unión al GTP/química , Subunidades beta de la Proteína de Unión al GTP/metabolismo , Expresión Génica , Datos de Secuencia Molecular , Sistemas de Lectura Abierta , Especificidad de Órganos/genética , Filogenia , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Conformación Proteica , Alineación de SecuenciaRESUMEN
We isolated two TATA-binding protein-associated factor (TAF) genes, TAF10 and TAF13, from pepper (Capsicum annuum). The complete coding sequences were amplified using reverse transcriptase-PCR on the basis of conserved sequence information of eggplant and several other plant species. Nucleotide sequence analysis of these two genes revealed that the pepper TAF10 gene encodes a protein of 103 amino acids that belongs to the TAF10 superfamily. The pepper TAF10 gene was highly expressed in the pericarp and placenta, moderately expressed in the stems, flowers, seeds and leaves, and weakly expressed in roots. The TAF13 gene was found to encode a protein of 130 amino acids that belongs to the TAF13 superfamily. The TAF13 gene was highly expressed in the stems, flowers and pericarp, moderately expressed in the leaves, placenta and seeds, and weakly expressed in roots.
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Capsicum/genética , Regulación de la Expresión Génica de las Plantas , Factores Asociados con la Proteína de Unión a TATA/genética , Secuencia de Aminoácidos , Secuencia de Bases , Clonación Molecular , Flores/metabolismo , Perfilación de la Expresión Génica , Datos de Secuencia Molecular , Hojas de la Planta/metabolismo , Proteínas de Plantas/biosíntesis , Proteínas de Plantas/genética , Raíces de Plantas/metabolismo , Tallos de la Planta/metabolismo , Proteínas del Complejo de Iniciación de Transcripción Pol1/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Semillas/metabolismo , Análisis de Secuencia de ADN , Factores Asociados con la Proteína de Unión a TATA/biosíntesis , Factores de Transcripción TFII/genéticaRESUMEN
We examined a possible relationship -420C>G SNP of the resistin gene with plasma resistin and C-reactive protein concentrations in intracerebral hemorrhage. Three hundred and forty-four Chinese Han patients with intracerebral hemorrhage and 344 age- and gender-matched healthy controls were included in our study. Plasma resistin and C-reactive concentrations were measured and SNP -420C>G was genotyped. The genotype frequencies in controls and patients were not significantly different (P = 0.672). Plasma resistin and C-reactive protein levels were significantly different between the SNP -420C>G genotypes, even after adjustment for age, gender and body mass index. The common homozygote (C-C) had the lowest resistin and C-reactive protein plasma concentrations; the plasma resistin and C-reactive protein concentrations in the heterozygote (C-G) and the rare allele homozygote (G-G) did not differ significantly. Plasma resistin levels were significantly associated with plasma C-reactive protein level. We conclude that SNP -420C>G of the resistin gene could be involved in the inflammatory component of intracerebral hemorrhage through enhanced production of resistin.
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Pueblo Asiatico/genética , Hemorragia de los Ganglios Basales/genética , Proteína C-Reactiva/metabolismo , Etnicidad/genética , Polimorfismo de Nucleótido Simple/genética , Resistina/sangre , Resistina/genética , Anciano , Anciano de 80 o más Años , Hemorragia de los Ganglios Basales/sangre , China , Femenino , Frecuencia de los Genes/genética , Estudios de Asociación Genética , Predisposición Genética a la Enfermedad , Humanos , Masculino , Persona de Mediana EdadRESUMEN
NADP-dependent isocitrate dehydrogenase (NADP-ICDH) is an important enzyme involved in energy metabolism. The complete coding sequence of the pepper (Capsicum annuum) NADP-ICDH gene was amplified using a reverse transcriptase PCR based on the conserved sequence information of the tomato and other Solanaceae plants and known highly homologous pepper ESTs. Nucleotide sequence analysis revealed that the pepper NADP-ICDH gene encodes a protein of 415 amino acids that has high homology with the proteins of seven species, Solanum tuberosum (100%), Citrus limon (98%), Daucus carota (98%), Nicotiana tabacum (98%), Vitis vinifera (99%), Arabidopsis thaliana (97%), and Oryza sativa (98%). Tissue expression analysis demonstrated that the pepper NADP-ICDH gene is over expressed in flower, pericarp and seed, moderately in placenta, weakly in stem and leaf, hardly expressed in root. At the abortion stages, activities and expression levels of NADP-ICDH in anthers of a sterile line were strongly reduced, while those in an F(1) hybrid remained normal. Activities and expression levels of NADP-ICDH were too low to maintain balanced energy metabolism in the sterile line, which indicated that stable transcripts of NADP-ICDH are necessary to maintain energy metabolism at a normal level. When the restorer gene was transferred to the cytoplasmic male sterile line, activities and expression level of NADP-ICDH were regulated by the restorer gene and became stable. The restorer gene likely plays an important role in keeping the balance of the energy metabolism within normal levels during microspore development.
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Capsicum/enzimología , Capsicum/genética , Genes de Plantas/genética , Isocitrato Deshidrogenasa/genética , Infertilidad Vegetal/genética , Análisis de Secuencia de ADN , Secuencia de Aminoácidos , Secuencia de Bases , Clonación Molecular , ADN Complementario/genética , Evolución Molecular , Perfilación de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Isocitrato Deshidrogenasa/química , Isocitrato Deshidrogenasa/metabolismo , Modelos Moleculares , Datos de Secuencia Molecular , Especificidad de Órganos/genética , Filogenia , Estructura Secundaria de Proteína , Estructura Terciaria de Proteína , ARN Mensajero/genética , ARN Mensajero/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Alineación de Secuencia , Homología Estructural de ProteínaRESUMEN
Species-diagnostic anatomical characters of fleshflies are not known for most immature stages or even adults, and an existing key may be incomplete or difûcult for nonspecialists to use. The use of sarcophagids for PMI estimations has been greatly hampered by their highly similar morphological characters. DNA-based method can be used as a supplemental means of morphological method in identification of forensically important sarcophagid flies. However, relying solely on single DNA fragment for delimiting species is considered to be unreliable, especially when the fragment was small. Sequence data of selected regions of the cytochrome oxidase subunit two (COII) and 16S ribosomal RNA (16S rRNA) genes of the most important Chinese fleshfly taxa associated with cadavers are presented, which can be instrumental for implementation of the Chinese Sarcophagidae database. Phylogenetic analysis of the sequenced segments showed that all sarcophagid specimens were properly assigned into five species, which indicated the possibility of separation congeneric species with the short fragments.
Asunto(s)
Dermatoglifia del ADN/métodos , ADN Mitocondrial/genética , Entomología/métodos , Ciencias Forenses/métodos , Sarcofágidos/clasificación , Sarcofágidos/genética , Animales , China , ADN Ribosómico/química , ADN Ribosómico/genética , Complejo IV de Transporte de Electrones/genética , Filogenia , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADNRESUMEN
With the development of molecular identification, there has been a great deal of discussion about the feature of the mitochondrial DNA (mtDNA) fragments. Although longer fragments may minimize stochastic variation across taxa and be more likely to reflect broader patterns of nucleotide divergence, shorter fragments have many advantages, such as quick, easy and economical. Extensive application of long mtDNA segments for species identification cannot always be achieved as a result of constraints in time and money. In the present study, a molecular identification method involving the sequencing of a 272-bp 'barcode' fragment of the mitochondrial cytochrome oxidase subunit I (COI) gene from 55 specimens, representing 7 Chinese sarcophagid species from varying populations, was evaluated. Phylogenetic analysis of the sequenced segments showed that all sarcophagid specimens were properly assigned into seven species, which indicated the possibility of separation congeneric species with the short fragments. The results of this research will be instrumental for the implementation of the Chinese Sarcophagidae database.
Asunto(s)
Código de Barras del ADN Taxonómico/métodos , Entomología/métodos , Ciencias Forenses/métodos , Sarcofágidos/clasificación , Sarcofágidos/genética , Animales , China , ADN Mitocondrial/genética , Complejo IV de Transporte de Electrones/genética , Proteínas de Insectos/genética , Datos de Secuencia Molecular , Filogenia , Reacción en Cadena de la Polimerasa , Alineación de Secuencia , Análisis de Secuencia de ADN , Especificidad de la EspecieRESUMEN
Abstract. Species-diagnostic anatomical characters of fleshflies are not known for most immature stages or even adults, and an existing key may be incomplete or difûcult for nonspecialists to use. The use of sarcophagids for PMI estimations has been greatly hampered by their highly similar morphological characters. DNA-based method can be used as a supplemental means of morphological method in identification of forensically important sarcophagid flies. However, relying solely on single DNA fragment for delimiting species is considered to be unreliable, especially when the fragment was small. Sequence data of selected regions of the cytochrome oxidase subunit two (COII) and 16S ribosomal RNA (16SrRNA) genes of the most important Chinese fleshfly taxa associated with cadavers are presented, which can be instrumental for implementation of the Chinese Sarcophagidae database. Phylogenetic analysis of the sequenced segments showed that all sarcophagid specimens were properly assigned into five species, which indicated the possibility of separation congeneric species with the short fragments.
RESUMEN
beta-catenin has emerged as a key regulator of Wnt signaling pathway, which plays an important role in the development and progression of various cancers. Its accumulation in nucleus of the esophagus squamous epithelium might be the crucial step for the carcinogenesis of esophageal squamous cell carcinoma (ESCC). To detect the proteins correlated with beta-catenin function, we used the established cell lines of pGen-3-con (Eca109 cells transfected by control vector) and pGen-3-CTNNB1 (Eca109 cells transfected by beta-catenin siRNA) as cell models for further analysis. Two-dimensional gel electrophoresis technology was performed to separate the proteins of pGen-3-con and pGen-3-CTNNB1 cell lines, respectively. The differential protein spots were analyzed by software analysis, subjected to in-gel digestion, and identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Consequently, 13 differentially expressed proteins between the two cell lines were identified, of which 14-3-3sigma, prohibitin, and nm23-H1 were further verified by western blotting and quantitative real-time reverse transcriptase-polymerase chain reaction. Then, the tissue microarray and immunohistochemical analysis were employed to research their relationship in ESCC and their corresponding normal mucosa tissues. The upregulation of prohibitin or the downregulation of 14-3-3sigma and nm23-H1 proteins was significantly associated with the proliferation, invasion depth, and lymph node metastasis of ESCC. There were statistically significant correlations between the expression of beta-catenin and the three proteins. The results presented here might provide potential protein markers to elucidate the mechanism of beta-catenin-mediated biologic characteristics for ESCC.
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Carcinoma de Células Escamosas/patología , Neoplasias Esofágicas/patología , Proteoma/análisis , beta Catenina/análisis , Proteínas 14-3-3/análisis , Proteínas 14-3-3/genética , Biomarcadores de Tumor/análisis , Biomarcadores de Tumor/genética , Western Blotting , Carcinoma de Células Escamosas/genética , Línea Celular Tumoral , Proliferación Celular , Progresión de la Enfermedad , Regulación hacia Abajo , Electroforesis en Gel Bidimensional , Neoplasias Esofágicas/genética , Esófago/citología , Exonucleasas/análisis , Exonucleasas/genética , Exorribonucleasas , Regulación Neoplásica de la Expresión Génica/genética , Humanos , Metástasis Linfática/patología , Nucleósido Difosfato Quinasas NM23/análisis , Nucleósido Difosfato Quinasas NM23/genética , Invasividad Neoplásica , Proteínas de Neoplasias/análisis , Proteínas de Neoplasias/genética , Prohibitinas , Análisis por Matrices de Proteínas , Proteínas Represoras/análisis , Proteínas Represoras/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Transfección , Regulación hacia Arriba , beta Catenina/genéticaRESUMEN
Accurate species identication is a crucial step in forensic entomology, as the insect collected on a corpse can provide useful information for estimation of postmortem interval (PMI). The utility of the forensically important Sarcophagidae (Diptera) for crime scene investigation has been severely restricted, as morphological identification is difficult, especially the identification of females and larvae. In this study, a method for using mitochondrial DNA (mtDNA) sequence data and phylogenetic analysis was performed to distinguish the three Chinese sarcophagid species: Boerttcherisca peregrina (Robineau-Desvoidy, 1830) Parasarcophaga albiceps (Meigen, 1826) and Parasarcophaga dux (Thompson, 1869) (Diptera: Sarcophagidae). DNA was extracted and analyzed by a 189 bp fragment of cytochrome coxidase subunit II (COII) gene. The monophyletic branches of the phylogenetic tree reveal that this marker is suitable for discrimination between these species, providing high support for separation on congeneric species. Therefore, the molecular method applied to the sarcophagid species identification is feasible.
Asunto(s)
Complejo IV de Transporte de Electrones/genética , Entomología/métodos , Ciencias Forenses/métodos , Proteínas de Insectos/genética , Sarcofágidos/clasificación , Sarcofágidos/genética , Animales , China , ADN Mitocondrial/genética , Datos de Secuencia Molecular , Filogenia , Análisis de Secuencia de ADNRESUMEN
Atrial secretion of atrial natriuretic peptide (ANP) has been shown to be regulated by atrial workload. Although modulating factors for the secretion of ANP have been reported, the role for intracellular Ca(2+) on the secretion of ANP has been controversial. The purpose of the present study was to define roles for L- and T-type Ca(2+) channels in the regulation of ANP secretion in perfused beating rabbit atria. BAY K 8644 (BAY K) increased atrial stroke volume and pulse pressure. BAY K suppressed ANP secretion and ANP concentration in terms of extracellular fluid (ECF) translocation concomitantly with an increase in atrial dynamics. BAY K shifted the relationship between ANP secretion and ECF translocation downward and rightward. These results indicate that BAY K inhibits myocytic release of ANP. In the continuous presence of BAY K, diltiazem reversed the effects of BAY K. Diltiazem alone increased ANP secretion and ANP concentration along with a decrease in atrial dynamics. Diltiazem shifted relationships between ANP secretion and atrial stroke volume or ECF translocation leftward. The T-type Ca(2+) channel inhibitor mibefradil decreased atrial dynamics. Mibefradil inhibited ANP secretion and ANP concentration in contrast with the L-type Ca(2+) channel inhibitor. These results suggest that activation of L- and T-type Ca(2+) channels elicits opposite effects on atrial myocytic release of ANP.
Asunto(s)
Factor Natriurético Atrial/metabolismo , Canales de Calcio Tipo L/fisiología , Canales de Calcio Tipo T/fisiología , Miocardio/metabolismo , Ácido 3-piridinacarboxílico, 1,4-dihidro-2,6-dimetil-5-nitro-4-(2-(trifluorometil)fenil)-, Éster Metílico/farmacología , Animales , Secreciones Corporales/efectos de los fármacos , Agonistas de los Canales de Calcio/farmacología , Bloqueadores de los Canales de Calcio/farmacología , Diltiazem/farmacología , Espacio Extracelular/metabolismo , Atrios Cardíacos/metabolismo , Técnicas In Vitro , Mibefradil/farmacología , Contracción Miocárdica , Conejos , Volumen Sistólico/efectos de los fármacosRESUMEN
OBJECTIVE: To investigate the effect of transforming growth factor beta 1 (TGF beta 1) and p27kip1 in gastric mucosa carcinogenesis. METHODS: RT-PCR was used to detect TGF beta 1 and p27kip1 mRNAs in normal gastric mucosa, simple hyperplasia, dysplasia and gastric carcinoma tissues respectively. RESULTS: There were expressions of TGF beta 1 and p27kip1 mRNAs in normal gastric mucosa, simple hyperplasia, dysplasia and gastric carcinoma tissues, the expressive levels of TGF beta 1 and p27kip1 mRNAs decreased gradually among the four groups. The expressive level of TGF beta 1 and p27kip1 mRNAs in gastric carcinoma group were significantly lower than those in other three groups(P < 0.05). CONCLUSIONS: The data indicate that TGF beta 1 and p27kip1 may play an important role in the development and carcinogenesis of gastric carcinoma. Semi-quantification PCR technique is effective for detecting mRNA expressive level in tissues.