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OBJECTIVE: Treatment of bilateral vertebral artery dissecting aneurysms presenting with subarachnoid hemorrhage remains challenging as bilateral deconstructive procedures may not be feasible. In this case series, we describe our approach to their management and review the pertinent literature. METHOD: A retrospective review of our prospectively collected database on aneurysms was performed to identify all patients with acute subarachnoid hemorrhage in the setting of bilateral intradural vertebral artery dissections (VAD) encompassing a period from January 2000 and March 2012. RESULT: Four patients (M/F = 2/2; mean age, 51.5 years) were identified. In two cases the site of rupture could be identified by angiographic and cross-sectional features; in these patients deconstructive treatment (proximal obliteration or trapping) of the ruptured site and reconstructive treatment of the unruptured site (using stents and coils) were performed. In the patients in whom the site of hemorrhage could not be determined, bilateral reconstructive treatment was performed. No treatment-related complications were encountered. Modified Rankin scale scores were 0-1 at discharge, and on follow-up (mean 63 months), no recurrence, in-stent thrombosis or new neurological deficits were encountered. CONCLUSION: We believe that single-stage treatment in patients with bilateral VAD is indicated: If the site of hemorrhage can be determined, we prefer deconstructive treatment on the affected site and reconstructive treatment on the non-affected site to prevent increased hemodynamic stress on the unruptured but diseased wall. If the site of dissection cannot be determined, we prefer bilateral reconstructive treatment to avoid increasing hemodynamic stress on the potentially untreated acute hemorrhagic dissection.
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Procedimientos Endovasculares/métodos , Hemorragia Subaracnoidea/cirugía , Disección de la Arteria Vertebral/cirugía , Adulto , Anatomía Transversal , Aneurisma Roto , Angiografía Cerebral , Bases de Datos Factuales , Procedimientos Endovasculares/efectos adversos , Femenino , Estudios de Seguimiento , Humanos , Masculino , Persona de Mediana Edad , Procedimientos de Cirugía Plástica , Estudios Retrospectivos , Stents , Hemorragia Subaracnoidea/etiología , Hemorragia Subaracnoidea/patología , Resultado del Tratamiento , Disección de la Arteria Vertebral/complicaciones , Disección de la Arteria Vertebral/patologíaRESUMEN
PURPOSE: The treatment of ruptured vertebral artery dissecting aneurysms (VADAs) continues to be controversial. Our goal was to evaluate the safety, efficacy, and long-term outcomes of internal trapping and stent-assisted coiling (SAC) for ruptured VADAs distal to the posterior inferior cerebellar artery (supra-PICA VADAs), which is the most common subset. METHODS: A retrospective review was conducted of 39 consecutive ruptured supra-PICA VADAs treated with internal trapping (n = 20) or with SAC (n = 19) at our institution. The clinical and angiographic data were retrospectively compared. RESULTS: The immediate total occlusion rate of the VADAs was 80 % in the trapping group, which improved to 88.9 % at the follow-ups (45 months on average). Unwanted occlusions of the posterior inferior cerebellar artery (PICA) were detected in three trapped cases. Incomplete obliteration of the VADA or unwanted occlusions of the PICA were detected primarily in the VADAs closest to the PICA. In the stenting group, the immediate total occlusion rate was 47.4 %, which improved to 100 % at the follow-ups (39 months on average). The immediate total occlusion rate of the VADAs was higher in the trapping group (p < 0.05), but the later total occlusion was slightly higher in the stenting group (p > 0.05). CONCLUSIONS: Our preliminary results showed that internal trapping and stent-assisted coiling are both technically feasible for treating ruptured supra-PICA VADAs. Although not statistically significant, procedural related complications occurred more frequently in the trapping group. When the VADAs are close to the PICA, we suggest that the lesions should be treated using SAC.
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Aneurisma Roto/terapia , Embolización Terapéutica , Aneurisma Intracraneal/terapia , Stents , Disección de la Arteria Vertebral/terapia , Adulto , Aneurisma Roto/diagnóstico por imagen , Angiografía Cerebral , Arterias Cerebrales/diagnóstico por imagen , Femenino , Estudios de Seguimiento , Humanos , Aneurisma Intracraneal/diagnóstico por imagen , Masculino , Persona de Mediana Edad , Estudios Retrospectivos , Rotura Espontánea , Resultado del Tratamiento , Arteria Vertebral/diagnóstico por imagen , Disección de la Arteria Vertebral/diagnóstico por imagenRESUMEN
OBJECTIVES: We aimed to evaluate risk factors of sis-VADAs recurrence after reconstructive treatment based on 113 reconstructed lesions. METHODS: A total of 111 patients (M:F = 68:43; median age, 47 years) with 113 sis-VADAs underwent reconstruction from October 2000 to March 2011, using stent(s) and coils. Treatments and predictors of recurrence were retrospectively analysed. RESULTS: Fifty-eight sis-VADAs underwent single-stent treatment, and the remaining 55 sis-VADAs underwent treatment with 2-4 overlapping stents. Follow-up angiography was available for 94 sis-VADAs 12 -78 months, with recurrence in ten patients, including seven angiographic recurrences and three post-treatment haemorrhagic recurrences. A higher rate of post-treatment recurrence was observed in the single stent group than in the multiple stents group (p = 0.010). The interaction between stent (s) implantation and immediate occlusion degree (odds ratio [OR] =3.152; 95% confidence interval [CI], 1.293-7.686; p = 0.012), between stent (s) implantation and the posterior inferior cerebellar artery (PICA) involvement (OR = 4.607; 95% CI, 1.172-18.113; p = 0.029), and between PICA involvement and immediate occlusion degree (OR = 5.018; 95% CI, 1.263-19.933; p = 0.022) affect recurrence in the reconstructed sis-VADAs. CONCLUSIONS: This single centre cohort study indicated that the interaction effect between stent (s) implantation, PICA involvement, and immediate occlusion degree were closely associated with recurrence after reconstructive treatment of sis-VADA. KEY POINTS: The interaction between stent (s) and immediate occlusion degree affect aneurysmal recurrence. The interaction between stent and PICA involvement affect aneurysmal recurrence. The interaction between PICA involvement and immediate occlusion degree affect aneurysmal recurrence.
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Embolización Terapéutica/instrumentación , Aneurisma Intracraneal/terapia , Procedimientos de Cirugía Plástica/métodos , Arteria Cerebral Posterior/patología , Stents , Disección de la Arteria Vertebral/terapia , Arteria Vertebral/diagnóstico por imagen , Adulto , Anciano , Prótesis Vascular , Angiografía Cerebral , Femenino , Estudios de Seguimiento , Humanos , Aneurisma Intracraneal/diagnóstico por imagen , Masculino , Persona de Mediana Edad , Arteria Cerebral Posterior/cirugía , Recurrencia , Estudios Retrospectivos , Arteria Vertebral/cirugía , Disección de la Arteria Vertebral/diagnóstico por imagenRESUMEN
To activate the expression of host genes that contribute to pathogen growth, pathogenic Xanthomonas bacteria inject their transcription activator-like effectors (TALEs) into plant cells and the TALEs bind to target gene promoters by the central repeat region consisting of near-perfect 34-amino-acid repeats (34-aa repeats). Based on the recognition codes between the 34-aa repeats and the targeted nucleotides, TALE-based technologies, such as designer TALEs (dTALEs) and TALE nucleases (TALENs), have been developed. Amazingly, every natural TALE invariantly has a truncated last half-repeat (LHR) at the end of the 34-aa repeats. Consequently, all the reported dTALEs and TALENs also harbour their LHRs. Here, we show that the LHRs in dTALEs are dispensable for the function of gene activation by both transient expression assays in Nicotiana benthamiana and gene-specific targeting in the rice genome, indicating that TALEs might originate from a single progenitor. In the light of this finding, we demonstrate that dTALEs can be constructed through two simple steps. Moreover, the activation strengths of dTALEs lacking the LHR are comparable with those of dTALEs harbouring the LHR. Our results provide new insights into the origin of natural TALEs, and will facilitate the simplification of the design and assembly of TALE-based tools, such as dTALEs and TALENs, in the near future.
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Proteínas Bacterianas/metabolismo , Secuencias Repetitivas de Aminoácido , Transactivadores/metabolismo , Secuencia de Aminoácidos , Proteínas Bacterianas/química , Genoma de Planta , Glucuronidasa/genética , Datos de Secuencia Molecular , Oryza/genética , Homología de Secuencia de Aminoácido , Nicotiana/enzimología , Nicotiana/microbiología , Transactivadores/química , Xanthomonas/metabolismo , Xanthomonas/patogenicidadRESUMEN
Chitinases are a group of pathogenesis-related proteins. The Brassica juncea chitinase gene BjCHI1 is highly inducible by pathogenic fungal infection, suggesting that the promoter of BjCHI1 might contain specific cis-acting element responsive to fungal attack. To identify the fungus-responsive element in BjCHI1 promoter (BjC-P), a series of binary plant transformation vectors were constructed by fusing the BjC-P or its deletion-derivatives to ß-glucuronidase (GUS) reporter gene. Expression of the GUS reporter gene was systematically assayed by a transient gene expression system in Nicotiana benthamiana leaves treated with fungal elicitor Hexa-N-Acetyl-Chitohexaose, as well as in transgenic Arabidopsis plants inoculated with fungus Botrytis cinerea. The histochemical and quantitative GUS assays showed that the W-box-like element (GTAGTGACTCAT) in the region (-668 to -657) was necessary for the fungus-response, although there were another five W-box-like elements in BjC-P. In addition, gain-of-function analysis demonstrated that the fragment (-409 to -337) coupled to the W-box-like element was needed for full magnitude of the fungal induction. These results revealed the existence of a novel regulation mechanism of W-box-like element involved in plant pathogenic resistance, and will benefit the potential application of BjC-P in engineering crops.
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Quitinasas/genética , Resistencia a la Enfermedad/genética , Regulación de la Expresión Génica de las Plantas , Genes de Plantas , Planta de la Mostaza/genética , Regiones Promotoras Genéticas , Secuencias Reguladoras de Ácidos Nucleicos , Arabidopsis/genética , Arabidopsis/microbiología , Botrytis , Quitinasas/metabolismo , Genes Reporteros , Glucuronidasa/genética , Planta de la Mostaza/metabolismo , Planta de la Mostaza/microbiología , Oligosacáridos/farmacología , Enfermedades de las Plantas/genética , Enfermedades de las Plantas/microbiología , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Modificadas Genéticamente , Nicotiana/efectos de los fármacos , Nicotiana/genéticaRESUMEN
Bacterial blight (BB), caused by Xanthomonas oryzae pv. oryzae (Xoo), is not only a disease devastating rice production worldwide, but also an ideal model system for the study of the interaction between plants and their bacterial pathogens. The rice near-isogenic line (NIL) CBB23, derived from a cross between a wild rice Oryza rufipogon accession (RBB16) and a susceptible indica rice variety (Jingang 30), is highly resistant to all field Xoo strains tested so far. Although the BB resistance of CBB23 has been widely used in rice breeding programmes, the mechanism of its extremely broad-spectrum resistance remains unknown. Here, we report the molecular cloning of an avirulence gene, designated as avrXa23, from Xoo strain PXO99(A) . We validate that AvrXa23, a novel transcription activator-like effector, specifically triggers the broad-spectrum BB resistance in CBB23. The prevalence of avrXa23 in all 38 Xoo strains surveyed may explain the broad-spectrum feature of BB resistance in CBB23. The results will significantly facilitate the molecular cloning of the corresponding resistance (R) gene in the host, and provide new insights into our understanding of the molecular mechanism for broad-spectrum disease resistance in plants.
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Proteínas Bacterianas/metabolismo , Oryza/metabolismo , Oryza/parasitología , Enfermedades de las Plantas/microbiología , Xanthomonas/patogenicidad , Regulación de la Expresión Génica de las Plantas , Datos de Secuencia MolecularRESUMEN
INTRODUCTION: Few studies focused on predictors of unfavorable outcomes (modified Rankin Scale, 2-6) after reconstructive treatment of the ruptured intracranial spontaneous vertebral artery dissection aneurysms (ris-VADAs), which was evaluated based on 57 reconstructed lesions in this study. METHODS: Results of 57 consecutive patients (M:Fâ=â29â¶28; median age, 48 years; range, 27 to 69 years) harboring 57 ris-VADAs, which were treated with coils combined with single stent(nâ=â32), double overlapping stents (nâ=â16), and triple overlapping stents (nâ=â9) between October 2000 to March 2011, were retrospectively reviewed and analyzed. RESULTS: The available (nâ=â54) mean durations of angiographic and clinical follow-ups were 27 months (range, 12 to 78) and 62 months (range, 12 to 132), respectively. The involvement of PICA (pâ=â0.004), size of lesions (pâ=â0.000), quantity of stent (pâ=â0.001), and coil type (pâ=â0.002) affected the immediate obliteration grade, which was only risk factor for angiographic recurrences (pâ=â0.031). Although the post-treatment outcomes did not differ between single stent and multiple stents (pâ=â0.434), 5 angiographic recurrences, 1 rebleeding and 1 suspected rebleeding, all occurred in partial obliteration after single-stent-assisted coiling. Progressive thrombosis and in-stent obliteration were not detected on follow-up angiograms. Older age (odds ratio [OR]â=â1.090; 95% confidence interval [CI], 1.004-1.184; pâ=â0.040) and unfavorable Hunt-Hess scale (ORâ=â4.289; 95%CI, 1.232-14.933; pâ=â0.022) were independent predictors of unfavorable outcomes in the reconstructed ris-VADAs. CONCLUSIONS: Immediate obliteration grade was only risk factor for angiographic recurrence after reconstructive treatment. Unfavorable Hunt-Hess grade and older age were independent predictors of unfavorable outcomes in ris-VADAs.
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Aneurisma Roto/cirugía , Procedimientos de Cirugía Plástica , Disección de la Arteria Vertebral/cirugía , Adulto , Anciano , Angiografía , Femenino , Estudios de Seguimiento , Humanos , Masculino , Persona de Mediana Edad , Estudios Retrospectivos , Resultado del Tratamiento , Disección de la Arteria Vertebral/diagnóstico por imagenRESUMEN
INTRODUCTION: We aimed to evaluate the feasibility, safety, efficacy, and predictors for outcome of reconstructive treatment with Solitaire™ AB stent(s) based on 54 cases of saccular aneurysms and 14 of acute symptomatic dissecting aneurysms. METHODS: Fifty-eight consecutive patients (M/Fâ=â28/30; median age, 53 years) harbouring 68 aneurysms (ruptured/unrupturedâ=â12/56) underwent treatment with Solitaire™ AB stent(s) implantation between April 2010 and August 2011 in our institution. The data were retrospectively reviewed and analysed. RESULTS: The technical success rate of Solitaire™ AB stenting was 100%. The rates of the overall and the treatment-related adverse events were 9% (6/68) and 6% (4/68), respectively, and the recurrent rate was 1% (1/68). All of the adverse events (nâ=â6) occurred in tiny (nâ=â1, ≤3 mm) or small (nâ=â5, >3 to ≤10 mm) aneurysms. The majority (75%, 3/4) of thromboembolic events (thrombus, nâ=â2; infarction, nâ=â2) occurred in ruptured lesions, and 2 intraprocedural aneurysm ruptures occurred in the course of coiling when the stent(s) was/were applied within 6 months. Subarachnoid haemorrhages (SAH, p<0.05) and immediate occlusion grades (p<0.05) were predictors for overall adverse events by univariate analysis. Compared with the immediate post-treatment angiographic results, the follow-up angiographic imaging (mean, 13 months; range, 6-25 months) revealed that stent(s) implantation enhanced the rate of class I occlusion from 34% (23/68) to 93% (63/68). SAH was the only predictor for unfavourable outcomes (the modified Rankin Scale score [mRS], 2-6) during the mean 19-month (range, 12-27 months) of clinical follow-ups (p<0.05). CONCLUSIONS: Although the complete obliteration of tiny and small aneurysms without complications remains a challenge, stent(s) implantation could lead to further occlusion of incompletely coiled aneurysms. SAH and the occlusion grade were the primary predictors for adverse events. SAH was the only predictor for unfavourable outcomes by univariate analysis.
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Aneurisma Intracraneal/cirugía , Stents , Adolescente , Adulto , Anciano , Angiografía , Femenino , Humanos , Aneurisma Intracraneal/patología , Masculino , Persona de Mediana Edad , Estudios Retrospectivos , Stents/efectos adversos , Adulto JovenRESUMEN
Accumulating evidence suggests that extracellular matrix (ECM) remodeling plays a significant role following acute ischemic stroke (AIS). Decorin (DCN) is a well-recognized molecule present in the ECM; however, the role of DCN in AIS remains unknown. The present study aimed to investigate whether plasma concentrations of DCN are altered in patients following an AIS and whether they are correlated with matrix metalloproteinase-2 (MMP-2) levels and other laboratory and clinical variables. Plasma concentrations of DCN were assessed in 102 patients with AIS (less than 7 days) and 120 control subjects using ELISA assays. The correlation between DCN concentrations and MMP-2 levels, Trial of Org 10172 in Acute Stroke Treatment (TOAST) subtypes, stroke severity and risk factors were evaluated. The expression of DCN was significantly decreased in patients with AIS (P<0.001), particularly in the large-artery atherosclerosis (LAAS) group. The levels of DCN were positively correlated with MMP-2 (R=0.332; P<0.001), thus MMP-2 is an independent predictor of DCN concentration (P<0.001). DCN levels below 8,500 pg/ml had sensitivity and specificity values of AIS of 79.4 and 62.8%, respectively and DCN below 8,500 pg/ml was associated with AIS (OR=4.8; 95% CI: 2.1-11.1; P<0.001) following adjustment for potential confounders. In conclusion, for the first time, a reduction in DCN was detected in patients following AIS and these altered plasma concentrations were correlated with MMP-2. Larger studies are required to further investigate whether DCN is involved in the pathogenesis of ischemic stroke.
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Decorina/sangre , Metaloproteinasa 2 de la Matriz/sangre , Accidente Cerebrovascular/sangre , Anciano , Área Bajo la Curva , Matriz Extracelular/metabolismo , Femenino , Humanos , Imagen por Resonancia Magnética , Masculino , Persona de Mediana Edad , Curva ROC , Factores de Riesgo , Índice de Severidad de la Enfermedad , Accidente Cerebrovascular/diagnóstico , Accidente Cerebrovascular/etiología , Tomografía Computarizada por Rayos XRESUMEN
Ischemic stroke is a common neurological disease and causes severe disability and death worldwide. Lysyl oxidase (LOX) plays a crucial role in the maintenance of extracellular matrix stability and may participate in vascular remodeling in the development of ischemic stroke. The objective of this study is to identify polymorphisms in LOX genes and investigate the association between LOX polymorphisms and the susceptibility to ischemic stroke in the Chinese population. Genomic DNA sequencing analysis was performed on all 7 exons and all exon/intron splice sites of lysyl oxidase and 850 bp upstream, including the predicted promoter region in 25 control subjects. The identified polymorphisms were then detected by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in 702 ischemic stroke cases and 733 age-matched controls. Data were analyzed using the Chi-square test. Two polymorphisms in the LOX gene, 473G/A (rs1800449) and rs2278226, were observed in the Chinese population. Frequencies of LOX 473AA genotype and A allele were significantly higher in ischemic stroke patients than in controls (odds ration (OR) = 1.76, 95 % confidence interval (CI) 1.16-2.67, P = 0.007; and OR = 1.33, 95 % CI 1.10-1.60, P = 0.003). Also, the prevalence of AC haplotype was significantly increased in ischemic stroke cases (OR = 1.32, 95 % CI 1.10-1.60, P = 0.004). Our data suggest that the G473A polymorphism of LOX gene could be a new risk factor for ischemic stroke.
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Isquemia Encefálica/genética , Polimorfismo de Nucleótido Simple , Proteína-Lisina 6-Oxidasa/genética , Accidente Cerebrovascular/genética , Isquemia Encefálica/enzimología , Estudios de Casos y Controles , Femenino , Frecuencia de los Genes , Estudios de Asociación Genética , Humanos , Desequilibrio de Ligamiento , Masculino , Persona de Mediana Edad , Oportunidad Relativa , Polimorfismo de Longitud del Fragmento de Restricción , Análisis de Secuencia de ADN , Accidente Cerebrovascular/enzimologíaAsunto(s)
Metaloproteinasa 9 de la Matriz/sangre , Neoplasias Pancreáticas/sangre , Inhibidor Tisular de Metaloproteinasa-1/sangre , Biomarcadores de Tumor/sangre , Recolección de Muestras de Sangre/métodos , Humanos , Neoplasias Pancreáticas/diagnóstico , Reproducibilidad de los Resultados , Sensibilidad y EspecificidadRESUMEN
OBJECTIVE: The formula 1/2abc was suitable for the estimation of the epidural, acute subdural, most intracerebral and even some chronic subdural hematoma volume, but its accuracy was prone to be affected by some particular shapes of hematomas. In this study, we aimed to validate a more accurate method for the estimation of the epidural, subdural and intracerebral hematoma volume. METHODS: In the present study, hematomas were categorized into the epidural, subdural and intracerebral groups, and every hematoma in three subgroups was respectively estimated with the following methods, including the computer-assisted planimetric analysis (gold standard), 2/3Sh, 1/2abc, 1/3abc and Tada's formula. Moreover, the percentage deviation of the estimated hematoma volume by every method in comparison with gold standard was assessed respectively. RESULTS: Compared with gold standard (39.59 ± 24.47 ml), the formula 2/3Sh (40.55 ± 26.58 ml, P > 0.05) overestimated the epidural hematoma volume by nearly 1% (0.01 ± 0.09, P > 0.05); similarly, the formula 2/3Sh (70.56 ± 49.06 ml, P > 0.05) in comparison with gold standard (69.98 ± 45.99 ml) overestimated the subdural hematoma volume by nearly <1% (-0.00 ± 0.13, P > 0.05). In addition, the intracerebral hematoma volume assessed by the formula 2/3Sh in contrast with gold standard (33.33 ± 33.19 ml) was 33.88 ± 34.39 ml (P > 0.05) with an only 2% (-0.02 ± 0.15, P > 0.05) underestimation. DISCUSSION: The formula 2/3Sh in contrast with other methods was a precise, simple, convenient estimation method, which was suitable for the epidural, subdural and intracerebral hematoma volume.
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Diagnóstico por Computador/métodos , Hematoma/clasificación , Hematoma/diagnóstico , Programas Informáticos , Tomografía Computarizada por Rayos X/métodos , Análisis de Varianza , Hemorragia Cerebral/diagnóstico , Estudios de Cohortes , Femenino , Hematoma Epidural Craneal/diagnóstico , Hematoma Subdural/diagnóstico , Humanos , MasculinoRESUMEN
OBJECTIVES: The formula 1/2abc was a relatively reliable technique for the estimation of mostly intracranial hematoma volume, but its accuracy was prone to be affected by some particular shapes of hematoma, such as multinodular, separated and so on. In this work, we aimed to validate a more accurate method for the estimation of intracranial hematoma volume, which were free limitations of axial slices shapes. METHODS: We investigated the computed tomography (CT) scans of 186 patients with intracranial hematoma. The volume values of 186 intracranial hematomas were respectively assessed using five different methods. RESULTS: For the formula 2/3Sh, the excellent correlation coefficient obtained using Pearson and Spearman's coefficients were 0.990 and 0.990, respectively. Meanwhile, the estimated intracranial hematoma volume using the formula 2/3Sh was fairly precise, with a <1% (-0.00+/-0.14, p>0.05) underestimation. However, the intracranial hematoma volume values were estimated by the formulas 1/2abc, Tada and 1/3abc, in comparison with the gold standard (43.16+/-37.55 ml), were 45.98+/-39.97 ml (p>0.05) with a 5% (0.05+/-0.25, p<0.05) overestimation, 48.12+/-41.84 ml (p>0.05) with 10% (0.10+/-0.26, p<0.05) overestimation and 30.65+/-26.65 ml (p<0.05) with a nearly 29% (-0.29+/-0.16, p<0.05) underestimation, respectively. DISCUSSIONS: Owing to the area, length and width of the largest axial hematoma slice clearly marked in CT imaging, the formula 2/3Sh in comparison with other methods was a precise, simple and convenient estimation technique.
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Encéfalo/diagnóstico por imagen , Hemorragias Intracraneales/diagnóstico por imagen , Interpretación de Imagen Radiográfica Asistida por Computador/métodos , Tomografía Computarizada por Rayos X/métodos , Análisis de Varianza , Humanos , Imagenología Tridimensional , Modelos Estadísticos , Estadísticas no ParamétricasRESUMEN
We have previously isolated a Brassica juncea cDNA encoding a novel chitinase BjCHI1 with two chitin-binding domains (Zhao and Chye in Plant Mol Biol 40:1009-1018, 1999). The expression of BjCHI1 was highly inducible by methyl jasmonate (MeJA) treatment, wounding, caterpillar feeding, and pathogenic fungal infection. These observations suggest that the promoter of BjCHI1 gene might contain specific cis-acting elements for stress responses. Here, we report the cloning and characterization of the BjCHI1 promoter. A 1,098 bp BjCHI1 genomic DNA fragment upstream of the ATG start codon was isolated by PCR walking and various constructs were made by fusing the BjCHI1 promoter or its derivatives to beta-glucuronidase reporter gene. The transgenic Arabidopsis plants showed that the BjCHI1 promoter responded to wounding and MeJA treatment, and to treatments with either NaCl or polyethyleneglycol (PEG 6000), indicating that the BjCHI1 promoter responses to both biotic and abiotic stresses. A transient gene expression system of Nicotiana benthamiana leaves was adopted for promoter deletion analysis, and the results showed that a 76 bp region from -695 to -620 in the BjCHI1 promoter was necessary for MeJA-responsive expression. Furthermore, removal of a conserved T/G-box (AACGTG) at -353 to -348 of the promoter greatly reduced the induction by MeJA. This is the first T/G-box element identified in a chitinase gene promoter. Gain-of-function analysis demonstrated that the cis-acting element present in the 76 bp region requires coupling with the T/G-box to confer full magnitude of BjCHI1 induction by MeJA.
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Brassica/genética , Quitinasas/genética , Proteínas de Plantas/genética , Regiones Promotoras Genéticas/genética , Acetatos/farmacología , Secuencia de Bases , Brassica/enzimología , Clonación Molecular , Ciclopentanos/farmacología , ADN de Plantas/química , ADN de Plantas/genética , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Glucuronidasa/genética , Glucuronidasa/metabolismo , Datos de Secuencia Molecular , Oxilipinas/farmacología , Reguladores del Crecimiento de las Plantas/farmacología , Polietilenglicoles/farmacología , Secuencias Reguladoras de Ácidos Nucleicos/genética , Elementos de Respuesta/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Análisis de Secuencia de ADN , Cloruro de Sodio/farmacología , Estrés Mecánico , Sitio de Iniciación de la TranscripciónRESUMEN
G-proteins (guanine nucleotide-binding proteins that usually exhibit GTPase activities) and related signal transduction processes play important roles in mediating plant defense responses; here, a rice (Oryza sativa) cDNA clone, OsGAP1, encoding a GTPase-activating protein (GAP) that also contains a protein kinase C conserved region 2 (C2) domain is reported. An interacting G-protein partner for the OsGAP1 protein was identified by yeast two-hybrid library screening and confirmed by co-immunoprecipitation; the GTPase-activation activity of OsGAP1 on this interacting G-protein was demonstrated using in vitro assays. OsGAP1 was induced by wounding in rice and the presence of the R locus Xa14 enhances such induction. Gain-of-function tests in transgenic rice and Arabidopsis thaliana showed that constitutive expression of OsGAP1 led to increased resistance to bacterial pathogens in both monocots and dicots.
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Proteínas Activadoras de GTPasa/metabolismo , Regulación de la Expresión Génica de las Plantas/fisiología , Oryza/metabolismo , Proteínas de Plantas/metabolismo , Secuencia de Aminoácidos , Arabidopsis/metabolismo , Clonación Molecular , Proteínas Activadoras de GTPasa/genética , Datos de Secuencia Molecular , Enfermedades de las Plantas , Proteínas de Plantas/genética , Técnicas del Sistema de Dos HíbridosRESUMEN
A cDNA clone (OsRHC1) was obtained, which encodes a novel RING zinc finger protein sharing similar structural features (multiple transmembrane domains at the N-half; a unique RING zinc finger consensus Cys-X(2)-Cys-X(11)-Cys-X-His-X(3)-Cys-X(2)-Cys-X(6)-Cys-X(2)-Cys at the C terminus) to a group of closely related annotated proteins from both monocots and dicots. OsRHC1 was found to be localized on plasma membrane of rice cells and induced by wounding in rice lines containing Xa loci. Ecotopic expression of the OsRHC1 cDNA from rice (a monocot) in transgenic Arabidopsis thaliana (a dicot) enhanced the defence response toward Pseudomonas syringae pv. tomato DC3000, suggesting that OsRHC1 may confer broad-spectrum disease resistance. The protective effects were neutralized in the presence of MG132 or in an npr1-3 mutation background, indicating that the function of OsRHC1 is dependent on the ubiquitin-mediated protein degradation via the 26S proteasome and the presence of the key defence response regulator NPR1.
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Arabidopsis/microbiología , Interacciones Huésped-Patógeno/fisiología , Oryza/genética , Proteínas de Plantas/metabolismo , Pseudomonas syringae/fisiología , Secuencia de Aminoácidos , Arabidopsis/fisiología , Proteínas de Arabidopsis/metabolismo , Clonación Molecular , Expresión Génica , Datos de Secuencia Molecular , Enfermedades de las Plantas , Proteínas de Plantas/genética , Plantas Modificadas Genéticamente/microbiología , Plantas Modificadas Genéticamente/fisiología , Complejo de la Endopetidasa Proteasomal/metabolismo , Dominios RING FingerRESUMEN
OBJECTIVE: To study the optimum inclusion process of clolloid mill for borneolumsyntheticum from Huoxuezhitong Capsules. METHODS: The study was carried out with orthogonal design. The borneol content in inclusion complex, the utilization ratio of borneol, and synthesis grading were used as evaluating indicator to review the effects of 3 factors, including the proportion of borneolumsyntheticum to beta-cyclodextrin (mol: mol), milling time and water addition. RESULTS: The optimum process were borneolumsyntheticum: beta-cyclodextrin was 1:1, the milling time was 40 min, water addition was double. CONCLUSION: The optimum process is feasible, and fit for industrial production, with the advantage of high utilization ratio, shot time and low energy consumption.
Asunto(s)
Canfanos/análisis , Medicamentos Herbarios Chinos/química , Aceites Volátiles/análisis , Tecnología Farmacéutica/métodos , beta-Ciclodextrinas , Disponibilidad Biológica , Cápsulas , Cromatografía de Gases , Cromatografía en Capa Delgada , Portadores de Fármacos , Estabilidad de Medicamentos , Medicamentos Herbarios Chinos/análisis , Aceites Volátiles/química , Factores de Tiempo , Agua/administración & dosificaciónRESUMEN
OBJECTIVE: To study optimum inclusion process condition for volatile oil with beta-cyclodextrin (beta-CD). METHODS: The optimum preparation condition were investigated by orthogonal design. The influence of utilization beta-CD and oil, inclusion temperature and time were studied to determin the utilization ratio of volatile oil and oil-bearing rate. RESULTS: The optimum preparation conditions were established as: volatile oil: beta-CD was 1:10, the inclusion temperature was 60 degrees C and inclusion time was 3h. CONCLUSION: The method is convenient and can he used for mass production.
Asunto(s)
Medicamentos Herbarios Chinos/química , Lamiaceae/química , Aceites Volátiles , Tecnología Farmacéutica/métodos , beta-Ciclodextrinas , Análisis de Varianza , Portadores de Fármacos , Combinación de Medicamentos , Estabilidad de Medicamentos , Medicamentos Herbarios Chinos/análisis , Aceites Volátiles/análisis , Aceites Volátiles/química , Aceites Volátiles/aislamiento & purificación , Plantas Medicinales/química , Temperatura , Factores de Tiempo , beta-Ciclodextrinas/químicaRESUMEN
Brassica juncea BjCHI1 is a unique chitinase with two chitin-binding domains. Here, we show that, unlike other chitinases, potato-expressed BjCHI1 shows hemagglutination ability. BjCHI1 expression in B. juncea seedlings is induced by Rhizoctonia solani infection, suggesting its protective role against this fungus. To verify this, transgenic potato (Solanum tuberosum L. cv. Desiree) plants expressing BjCHI1 generated by Agrobacterium-mediated transformation were challenged with R. solani. We also transformed potato with a cDNA encoding Hevea brasiliensis beta-1,3-glucanase, designated HbGLU, and a pBI121-derivative that contains cDNAs encoding both BjCHI1 and HbGLU. In vitro fungal bioassays using Trichoderma viride showed that extracts from transgenic potato lines co-expressing BjCHI1 and HbGLU inhibited fungal growth better than extracts from transgenic potato expressing either BjCHI1 or HbGLU, suggesting a synergistic effect. Consistently, in vivo fungal bioassays with soil-borne R. solani on young transgenic potato plants indicated that the co-expressing plants showed healthier root development than untransformed plants or those that expressed either BjCHI1 or HbGLU. Light microscopy and transmission electron microscopy revealed abundant intact R. solani hyphae and monilioid cells in untransformed roots and disintegrated fungus in the BjCHI1-expressing and the BjCHI1 and HbGLU co-expressing plants. Observations of collapsed epidermal cells in the co-expressing potato roots suggest that these proteins effectively degrade the fungal cell wall, producing elicitors that initiate other defense responses causing epidermal cell collapse that ultimately restricts further fungal penetration.