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The present study reports a rare case of Taenia saginata infection, which was initially diagnosed as acute cholecystitis in a Tibetan patient at the Qinghai-Tibetan Plateau pastoral area, China. A 45-year-old female was initially diagnosed with acute cholecystitis at a hospital in China. She had a slight fever, weight loss and constipation and complained of pain in the upper abdomen and left back areas. Increase of monocyte, eosinophil and basophil levels were shown. Taenia sp. eggs were detected in a fecal examination. An adult tapeworm approximately 146 cm in length, whitish-yellow color, was collected from the patient after treatment with traditional Chinese medicine. The adult tapeworm had a scolex and proglottids with genital pores. The scolex was rectangular shape with 4 suckers and rostellum without hooklet. The cox1 gene sequence shared 99.5-99.8% homology with that of T. saginata from other regions in China. The patient was diagnosed finally infected with T. saginata by morphological and molecular charateristics.
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Colecistitis Aguda , Taenia saginata , Taenia , Teniasis , Adulto , Animales , China , Errores Diagnósticos , Femenino , Humanos , Persona de Mediana Edad , Taenia/genética , Taenia saginata/genética , Teniasis/diagnóstico , TibetRESUMEN
Coenurosis is an important zoonotic helminthic disease caused by the larval stage of the tapeworm Taenia multiceps. This parasite typically infects the brain of the intermediate hosts, including sheep, goat, cattle and even humans. We report a case of T. multiceps infection in a yak confirmed by clinical symptoms, morphological characteristics, and molecular and phylogenetic analyses. The coenurus was thin-walled, whitish, and spherical in shape with a diameter of 10 cm. The parasite species was identified as T. multiceps by PCR amplification and sequencing of the 18S rRNA, cox1 and nad1 genes. Three gene sequences all showed high homology (all above 97%) with the reference sequences from different hosts. Moreover, phylogenetic reconstructions with the 3 published Taenia gene sequences confirmed that the Qinghai yak isolate was closely related to T. multiceps. Although there are advanced diagnosis and treatment methods for coenurosis, early infection is difficult to diagnose. Importantly, the findings of yak infection case should not be ignored due to its zoonotic potential.
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Enfermedades de los Bovinos/parasitología , Neurocisticercosis/veterinaria , Taenia/genética , Animales , Bovinos , Ciclooxigenasa 1/genética , Electroforesis en Gel de Agar/veterinaria , Masculino , NAD/genética , Neurocisticercosis/parasitología , Filogenia , Reacción en Cadena de la Polimerasa/veterinaria , ARN Ribosómico 18S/genética , Alineación de Secuencia/veterinaria , Análisis de Secuencia de ADN/veterinaria , Taenia/clasificación , Taenia/aislamiento & purificación , TibetRESUMEN
Six cystic metacestodes were found in the abdominal muscles of a wild rabbit, Lepus sinensis, in China. The coenurus contained one or more scolices armed with hooklets. Mitochondrial cox1 (1,623 bp) confirmed 98% homology with cox1 of Taenia serialis. This is the first report of T. serialis infection in an intermediate host in the Qinghai Tibetan Plateau Area, China.
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Animales Salvajes/parasitología , Infecciones por Cestodos/parasitología , Infecciones por Cestodos/veterinaria , Conejos/parasitología , Enfermedades de los Roedores/parasitología , Taenia/aislamiento & purificación , Taenia/patogenicidad , Animales , China , Masculino , Filogenia , Taenia/anatomía & histología , Taenia/genética , Tibet , Zoonosis/prevención & controlRESUMEN
BACKGROUND: Echinococcus multilocularis causes alveolar echinococcosis (AE) and is widely prevalent in Qinghai Province, China, where a number of different species have been identified as hosts. However, limited information is available on the Qinghai vole (Lasiopodomys fuscus), which is hyper endemic to Qinghai Province and may represent a potential intermediate host of E. multilocularis. Thus, L. fuscus could contribute to the endemicity of AE in the area. METHODS: Fifty Qinghai voles were captured from Jigzhi County in Qinghai Province for the clinical identification of E. multilocularis infection via anatomical examination. Hydatid fluid was collected from vesicles of the livers in suspected voles and subjected to a microscopic examination and PCR assay based on the barcoding gene of cox 1. PCR-amplified segments were sequenced for a phylogenetic analysis. E. multilocularis-infected Qinghai voles were morphologically identified and subjected to a phylogenetic analysis to confirm their identities. RESULTS: Seventeen of the 50 Qinghai voles had E. multilocularis-infection-like vesicles in their livers. Eleven out of the 17 Qinghai voles presented E. multilocularis infection, which was detected by PCR and sequencing. The phylogenetic analysis showed that all 11 positive samples belonged to the E. multilocularis Asian genotype. A morphological identification and phylogenetic analysis of the E. multilocularis-infected Qinghai voles confirmed that all captured animals were L. fuscus. CONCLUSIONS: L. fuscus can be infected with E. multilocularis and plays a potential role in the life cycle and epidemiology of E. multilocularis in the Qinghai-Tibetan Plateau of China.
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Arvicolinae , Reservorios de Enfermedades/veterinaria , Equinococosis/veterinaria , Echinococcus multilocularis/aislamiento & purificación , Enfermedades de los Roedores/epidemiología , Animales , Arvicolinae/clasificación , Arvicolinae/genética , China/epidemiología , Reservorios de Enfermedades/parasitología , Equinococosis/epidemiología , Equinococosis/parasitología , Equinococosis/transmisión , Echinococcus multilocularis/genética , Complejo IV de Transporte de Electrones/genética , Proteínas del Helminto/genética , Masculino , Filogenia , Reacción en Cadena de la Polimerasa/veterinaria , Prevalencia , Enfermedades de los Roedores/parasitología , Enfermedades de los Roedores/transmisiónRESUMEN
This study was carried out to determine the pathogen-causing diarrhoea in sheep Ovis aries in the Qinghai Tibetan Plateau Area, China. A trophozoite was identified as species of ciliate alveolates infecting the sheep based on morphological characteristics examined by microscope. It was mostly spherical, colourless and transparent, with many vesicles. Macronucleus and contractile vacuoles could not be distinguished. Size of the trophozoite was 80-180 × 70-150 µm and its surface was covered with cilia. Molecular analysis based on sequences of 18S rRNA and ITS genes confirmed the ciliate species as Balantidium coli. According to the literature, there have been many epidemiological investigations of B. coli infection in pigs, monkeys and humans. To our knowledge, this was the first report of B. coli infections in sheep in the Qinghai Tibetan Plateau Area of China, or eleswhere around the world. Importantly, the sheep case was rare but raised our concern that B. coli may spread across species and expand its host range.
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Balantidiasis/veterinaria , Balantidium/aislamiento & purificación , Diarrea/veterinaria , Enfermedades de las Ovejas/parasitología , Animales , Balantidiasis/parasitología , Balantidium/clasificación , Balantidium/citología , Balantidium/genética , Análisis por Conglomerados , ADN Protozoario/química , ADN Protozoario/genética , ADN Ribosómico/química , ADN Ribosómico/genética , ADN Espaciador Ribosómico/química , ADN Espaciador Ribosómico/genética , Diarrea/parasitología , Masculino , Microscopía , Filogenia , ARN Ribosómico 18S/genética , Análisis de Secuencia de ADN , Ovinos , Oveja Doméstica , TibetRESUMEN
AIM: To investigate clinical features of optic nerve sheath meningioma (ONSM) that was misdiagnosed, and to find methods to reduce the misdiagnoses. METHODS: Retrospective series study. Twenty-five misdisgnosed patients with unilateral ONSM were collected from Jan. 2008 to Jan. 2015 and the clinical records reviewed. RESULTS: Patients were misdiagnosed with acute papillitis most frequently (n=17), immediately followed by optic atrophy (n=8), ischemic optic neuropathy (n=5), acute retrobulbar optic neuritis (n=5), optic disc vasculitis (n=3). For each patient, the minimum frequency of misdiagnoses was once and the maximum was 4 times. As for the lasting time of being misdiagnosed, the shortest was 1.5mo and the longest was 45mo. Twenty-one cases (84%) were once treated with glucocorticoids, and its side effects was found in seventeen patients. Twenty patients (80%) complained with varying degree of vision loss. When a definite diagnosis was made, sixteen cases (64%) showed slight exophthalmos and eighteen cases (72%) had the tubular ONSM. CONCLUSION: ONSM without loss obvious exophthalmos is easily misdiagnosed in clinic, and for most of these ONSMs are tubular.
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AIM: To investigate the role of Brn-3b in differentiation process of stem cells derived from retinal Müller cells into the ganglion cell. METHODS: The passage culture method of Müller cells from retina of newborn Sprague Dawley rats was carried out by repeated incomplete pancreatic enzyme digestion method. The cells were detected by fluorescence-activated cell sorter (FACS), immunohistochemistry technology and reverse transcription-polymerase chain reaction (RT-PCR) to determine the purity. The third passage of cells was induced in the serum-free dedifferentiation medium. The expression of the specific markers Ki-67 and nestin of retinal stem cells was measured by RT-PCR and Western blot. The cell proliferation of retinal stem cells was detected by 5-ethynyl-2'-deoxyuridine (Edu) staining. The cells were randomly divided into 5 groups as follows: group A: Brn-3bsiRNA group; group B: Brn-3b control siRNA group; group C: pGC-Brn-3b-green fluorescent protein (GFP) group; group D: pGC-GFP group; group E: control group (without any handling). The purified Müller cells were cultured for 3-7d, then, the percentage of ganglion cells was counted by immunofluorescence staining. RESULTS: FACS demonstrated the purity of retinal Müller cells was more 97.44%. A few spherical cell spheres appeared. Immunofluorescence staining showed that stem cells within the spheres were positive for retinal stem cell-specific markers nestin (red fluorescence, 92.94%±6.48%) and Ki-67 (green fluorescence, 85.96%±6.04%). Meanwhile, RT-PCR analysis showed cell spheres in the culture to have expressed a battery of transcripts characteristic of stem cells such as nestin and Ki-67, which were absent in the Müller cells. Western blot analysis further confirmed the expression of nestin and Ki-67 in the cell spheres but not in the Müller cells. Edu staining showed most of the nuclei within the cell spheres were stained red (82.80%±6.65%), suggesting the new cell spheres had the capacity for effective proliferation. The statistics result showed the difference between Brn-3bsiRNA group and Brn-3b control siRNA group or the control group was significant (F=15, P<0.05), while the difference between Brn-3b control siRNA group or the control group was not statistically significant (P>0.05). CONCLUSION: The repeated incomplete pancreatic enzyme digestion method is an efficient and practical method to purify retinal Müller cells. Retinal stem cells were successfully cloned in the dedifferentiational medium. Retinal Müller cells are accessible sources of retinal stem cells. Brn-3b is an important regulatory gene in stem cells differentiated into retinal ganglion cell.
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Glaucoma is one of the leading eye diseases resulting in blindness due to the death of retinal ganglion cells. This study aimed to develop novel protocol to promote the differentiation of retinal Müller cells into ganglion cells in vivo in a rat model of glaucoma. The stem cells dedifferentiated from rat retinal Müller cells were randomized to receive transfection with empty lentivirus PGC-FU-GFP or lentivirus PGC-FU-Atoh7-GFP, or no transfection. The stem cells were induced further to differentiate. Ocular hypertension was induced using laser photocoagulation. The eyes were injected with Atoh7 expression vector lentivirus PGC-FU-Atoh7-GFP. Eyeball frozen sections, immunohistochemistry, RT-PCR, Western bolt, and apoptosis assay were performed. We found that the proportion of ganglion cells differentiated from Atoh7-tranfected stem cells was significantly higher than that of the other two groups. The mean intraocular pressure of glaucomatous eyes was elevated significantly compared with those of contralateral eyes. Some retinal Müller cells in the inner nuclear layer entered the mitotic cell cycle in rat chronic ocular hypertension glaucoma model. Atoh7 contributes to the differentiation of retinal Müller cells into retinal ganglion cells in rat model of glaucoma. In conclusion, Atoh7 promotes the differentiation of Müller cells-derived retinal stem cells into retinal ganglion cells in a rat model of glaucoma, thus opening up a new avenue for gene therapy and optic nerve regeneration in glaucoma.
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Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/fisiología , Diferenciación Celular/fisiología , Ganglios/citología , Glaucoma/patología , Retina/citología , Células Madre/citología , Animales , Secuencia de Bases , Western Blotting , Cartilla de ADN , Modelos Animales de Enfermedad , Ratas , Ratas Sprague-Dawley , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
KEY MESSAGE: TaUBA functions as a negative regulator of salt and drought stress response in transgenic Arabidopsis, either the UBA domain or the zinc finger domain is crucial for TaUBA's function. TaUBA (DQ211935), which is a UBA domain-containing protein in wheat, was cloned and functionally characterized. Southern blot suggested that TaUBA is a low copy gene in common wheat. qRT-PCR assay showed that the expression of TaUBA was strongly induced by salt and drought stress. When suffering from drought and salt stresses, lower proline content and much higher MDA content in the TaUBA overexpressors were observed than those of the wild-type control, suggesting TaUBA may function as a negative regulator of salt and drought stress response in plants. To study whether the UBA domain or the zinc finger domain affects the function of TaUBA, TaUBAΔUBA (deletion of UBA domain) and TaUBA-M (Cys464Gly and Cys467Gly) overexpression vectors were constructed and transformed into Arabidopsis. Upon drought and salt stresses, the TaUBAΔUBA-and TaUBA-M-overexpressed plants accumulated much more proline and lower MDA than the wild-type control, the TaUBA-overexpressors lost water more quickly than TaUBAΔUBA-and TaUBA-M-overexpressed plants as well as the wild-type control, suggesting that overexpression of TaUBAΔUBA or TaUBA-M improved the drought and salt tolerance of transgenic Arabidopsis plants and the possibility of ubiquitination role in the regulation of osmolyte synthesis and oxidative stress responses in mediating stress tolerance. qRT-PCR assay of stress-related genes in transgenic plants upon drought and salt stresses suggested that TaUBA may function through down-regulating some stress related-transcription factors and by regulating P5CSs to cope with osmotic stress.
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Regulación de la Expresión Génica de las Plantas , Proteínas de Plantas/metabolismo , Triticum/genética , Arabidopsis/genética , Arabidopsis/fisiología , Sequías , Proteínas de Plantas/genética , Plantas Modificadas Genéticamente , Estructura Terciaria de Proteína , Tolerancia a la Sal , Cloruro de Sodio/farmacología , Estrés Fisiológico , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Triticum/fisiologíaRESUMEN
INTRODUCTION: Retinal Müller cells exhibit the characteristics of retinal progenitor cells, and differentiate into ganglion cells under certain conditions. However, the number of ganglion cells differentiated from retinal Müller cells falls far short of therapeutic needs. This study aimed to develop a novel protocol to promote the differentiation of retinal Müller cells into ganglion cells and explore the underlying signaling mechanisms. METHODS: Müller cells were isolated and purified from rat retina and induced to dedifferentiate into retinal stem cells. Next the stem cells were transfected with lentivirus PGC-FU-GFP or lentivirus PGC-FU-Atoh7-GFP. In addition, the stem cells were transfected with Brn-3b siRNA or Isl-1 siRNA or treated with Notch inhibitor gamma-secretase inhibitor (GSI). RESULTS: The proportion of ganglion cells differentiated from Atoh7-tranfected stem cells was significantly higher than that of controls. Knockdown of Brn-3b or Isl-1 inhibited, while GSI promoted, the differentiation into retinal ganglion cells. Atoh7 promoted the expression of Brn-3b and Isl-1 but inhibited the expression of Notch1. CONCLUSIONS: Atoh7 promotes the differentiation of Müller cells-derived retinal stem cells into retinal ganglion cells by inhibiting Notch signaling, thus opening up a new avenue for gene therapy and optic nerve regeneration in glaucoma.
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Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/fisiología , Receptores Notch/fisiología , Células Ganglionares de la Retina/metabolismo , Células Madre/citología , Animales , Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/genética , Diferenciación Celular , Proliferación Celular , Regulación del Desarrollo de la Expresión Génica , Ratas , Receptores Notch/genética , Transducción de Señal , TransfecciónRESUMEN
AIM: To study two methods for culturing and purifying Sprague-Dawley (SD) rat retinal Müller cells and determine which one is better. METHODS: The passage culture method of Müller cells was respectively carried out by complete pancreatic enzyme digestion method and repeated incomplete pancreatic enzyme digestion method. After culturing retinal cells for one month through these two methods, fluorescence-activated cell sorter (FACS), RT-PCR, and immunohistochemistry technology were performed to examine the enrichment and purity of Müller glial cells, and carried out two-sample approximate t test using SSPS 13.0 to further compare the Müller cell positive rate in both methods. RESULTS: The statistical results showed that the purity of Müller cells was 83.2%±5.16% in group A, and the purity was 98.5%±1.08% in group B. The two-sample approximate t test analysis demonstrated that the difference between group A and group B was statistically significant (t=-9.178, P<0.005). The results clearly exhibited a difference between the purity of Müller cells cultured by the complete pancreatic enzyme digestion method (group A) and the repeated incomplete pancreatic enzyme digestion method (group B). CONCLUSION: Compared with the complete pancreatic enzyme digestion method, this novel method was more efficient and a higher purity of Müller cells could be obtained using this approach.
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The transmittance of one-dimensional photonic crystals consisting of superconductor and lossless dielectric has been systematically studied through the transfer-matrix method. Obviously, the shift of the photonic bandgap (PBG) becomes more noticeable by adjusting the thicknesses of the dielectric layers than that of superconductor layers. Furthermore, the number of PBGs can be controlled by varying the thicknesses of dielectric layers. Compared to the thicknesses of the dielectric layers, the width of the PBGs is more sensitive to the thicknesses of the superconductor layers. However, the width of the first PBG promptly varies when the thicknesses of the dielectric layers increase from 0 to 40 nm. If the contribution of the normal conducting electrons of the superconductor is nonnegligible, the temperature of the superconductor has no influence on the width of the PBGs. Moreover, the damp coefficient does not affect the PBGs under low-temperature conditions.
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F-box protein is an important subunit of SCF complex, an E3 ligase in ubiquitin system, and its function is determined through mediating the specific recognition and combining with substrate protein. TaFRA (F-box protein related to abiotic stress) was identified by RACE based on the fragments diferently expressing in wheat seedling exposed to salt stress and encodes an F-box protein. In this study, pBD-TaFRA bait expression vector was constructed, and cDNA+pGAD+pBD was directly co-transformed into yeast hybrid system to screen condidate proteins interacting with TaFRA. Fourty-four candidate proteins were obtained, in which 32 were known proteins and transcript factors related to stress tolerance such as thioredoxin, metallothinein, ATP synthase, and serine/threonine protein kinase etc. This indicates that TaFRA participates in stress response through regulating above condidate genes, which will provide basis for revealing the mechanism of TaFRA reaction to abiotic stress.
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Proteínas F-Box/metabolismo , Mapeo de Interacción de Proteínas , Estrés Fisiológico , Técnicas del Sistema de Dos Híbridos , Clonación Molecular , Proteínas F-Box/genética , Saccharomyces cerevisiae/genéticaRESUMEN
Glutamine synthetase (GS) plays a key role in the growth, nitrogen (N) use and yield potential of cereal crops. Investigating the haplotype variation of GS genes and its association with agronomic traits may provide useful information for improving wheat N-use efficiency and yield. We isolated the promoter and coding region sequences of the plastic glutamine synthetase isoform (GS2) genes located on chromosomes 2A, 2B and 2D in bread wheat. By analyzing nucleotide sequence variations of the coding region, two, six and two haplotypes were distinguished for TaGS2-A1 (a and b), TaGS2-B1 (a-f) and TaGS2-D1 (a and b), respectively. By analyzing the frequency data of different haplotypes and their association with N use and agronomic traits, four major and favorable TaGS2 haplotypes (A1b, B1a, B1b, D1a) were revealed. These favorable haplotypes may confer better seedling growth, better agronomic performance, and improved N uptake during vegetative growth or grain N concentration. Our data suggest that certain TaGS2 haplotypes may be valuable in breeding wheat varieties with improved agronomic performance and N-use efficiency.
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Pan , Glutamato-Amoníaco Ligasa/genética , Haplotipos/genética , Nitrógeno/metabolismo , Carácter Cuantitativo Heredable , Triticum/enzimología , Triticum/genética , Alelos , China , Cruzamientos Genéticos , Genes de Plantas/genética , Glutamato-Amoníaco Ligasa/metabolismo , Haploidia , Hidroponía , Endogamia , Isoenzimas/genética , Isoenzimas/metabolismo , Datos de Secuencia Molecular , Recombinación Genética/genética , Plantones/genética , Plantones/crecimiento & desarrollo , Homología de Secuencia de Ácido Nucleico , Triticum/crecimiento & desarrolloRESUMEN
To verify the genome components of Thinopyrum ponticum Liu alt; Wang and Th. intermedium [Host] Barkworth alt; Dewey, six specific primer pairs were designed according to the sequence of an autonomous centromeric retrotransposon of wheat (CRW) from Triticum boeoticum. Several DNA fragments were amplified by PCR from the genomic DNA of the diploid species Pseudoroegneria spicata A Löve. After sequence alignment, a 1.755 kb fragment was obtained and named as pStC1 (St genome centromere associated sequence, GenBank accession No. FJ952565). This fragment contained an 800 bp fragment highly homologous to the LTR region of autonomous CRW, a short fragment partial homology to the gag region of CRW, and an AGCAAC-rich tandem repeat. Fluorescent in situ hybridization (FISH) using pStC1 as a probe was carried out on the chromosomes of Th. ponticum, Th. intermedium and Triticum aestivum cv. Chinese Spring. Th. ponticum was proved to have two St and three E genomes (St1St2EeEbEx). In Th. intermedium, strong FISH signals were observed on St genome chromosomes, while faint signals were also found on some E-genome chromosomes at their pericentromeric regions. These results indicated that during speciation of sub-genomes in the allopolyploids of Thinopyrum genus, concerted evolution might have occurred at centromeric and pericentromeric regions.
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Hibridación Fluorescente in Situ/métodos , Poaceae/genética , Triticum/genética , Secuencia de Bases , Centrómero/genética , Evolución Molecular , Genoma de Planta/genética , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa , Retroelementos/genéticaRESUMEN
Deficiency of micronutrients, especially iron and zinc, has been a serious malnutrition problem worldwide in human health. Increasing Fe and Zn concentrations in grains by means of plant breeding is a sustainable, effective and important way to improve human mineral nutrition and health. However, little information on grain Fe and Zn concentrations in Chinese wheat genotypes is available. Therefore, to determine the nutrients status especially these of micronutrients in wheat grain is necessary and very useful. Two hundred sixty two genotypes were selected from the wheat mini-core collections, which contained 23090 wheat genotypes in China and represented 72.2% of total genetic variation. All 262 genotypes were grown in soils of similar geographical and climate location in order to minimize the environmental effect. After harvesting, the grains were washed with deionized water and dried (around 70 degrees C), then digested in HNO3 solution using a microwave accelerating reaction system (MARS). Nutrient concentrations in stock solution were analyzed by inductively coupled plasma atomic emission spectrometry (ICP-AES). Remarkable genetic variations among grain nutrient concentrations (Fe, Mn, Cu, Zn, Mg, Ca, K and P ) in the tested genotypes were detected. The concentrations of Fe, Zn, Mn, Cu, Ca, Mg, K and P in wheat grain were in the ranges of 34.2-61.2, 26.3-76.0, 20.9-56.7, 3.4-9.8, 290-976, 1129-2210 mg x kg(-1); 0.34%-0.85% and 0.296%-0.580%, respectively. The corresponding average values were 45.1, 50.2, 37.9, 6.5, 515, 1772 mg x kg(-1), 0.55% and 0.451%, respectively. Significant positive correlations between micronutrients (Fe, Mn, Zn, and Cu) in wheat grains were detected, and the correlation coefficients were 0.395** (Fe and Mn), 0.424** (Fe and Zn), 0.574** (Fe and Cu), and 0.474** (Mn and Cu), respectively. However, no significant difference was found in grain nutrient concentrations between spring-wheat and winter-wheat genotypes. This study provides valuable and important information for breeding wheat genotypes which are enriched with minerals in grains, especially Fe and Zn
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Extractos Vegetales/análisis , Espectrofotometría Atómica/métodos , Oligoelementos/análisis , Triticum/química , China , Valor NutritivoRESUMEN
The high molecular weight glutenin subunit (HMW-GS) pair 1Bx13 + 1By16are recognized to positively correlate with bread-making quality; however, their molecular data remain unknown. In order to reveal the mechanism by which 1By16 and 1Bx13 creates high quality, their open reading frames (ORFs) were amplified from common wheat Atlas66 and Jimai 20 using primers that were designed based on published sequences of HMW glutenin genes. The ORF of 1By16 was 2,220 bp, deduced into 738 amino acid residues with seven cysteines including 59 hexapeptides and 22 nanopeptides motifs. The ORF of 1Bx13 was 2,385 bp, deduced into 795 amino acid residues with four cysteines including 68 hexapeptides, 25 nanopeptides and six tripeptides motifs. We found that 1By16 was the largest y-type HMW glutenin gene described to date in common wheat. The 1By16 had 36 amino acid residues inserted in the central repetitive domain compared with 1By15. Expression in bacteria and western-blot tests confirmed that the sequence cloned was the ORF of HMW-GS 1By16, and that 1Bx13 was one of the largest 1Bx genes that have been described so far in common wheat, exhibiting a hexapeptide (PGQGQQ) insertion in the end of central repetitive domain compared with 1Bx7. A phylogenetic tree based on the deduced full-length amino acid sequence alignment of the published HMW-GS genes showed that the 1By16 was clustered with Glu-1B-2, and that the 1Bx13 was clustered with Glu-1B-1 alleles.
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Genes de Plantas , Glútenes/genética , Glútenes/aislamiento & purificación , Poliploidía , Subunidades de Proteína/genética , Subunidades de Proteína/aislamiento & purificación , Triticum/genética , Alelos , Secuencia de Aminoácidos , Electroforesis en Gel de Poliacrilamida , Glútenes/química , Interacciones Hidrofóbicas e Hidrofílicas , Datos de Secuencia Molecular , Peso Molecular , Sistemas de Lectura Abierta/genética , Filogenia , Reacción en Cadena de la Polimerasa , Estructura Secundaria de Proteína , Subunidades de Proteína/química , Alineación de Secuencia , Homología de Secuencia de AminoácidoRESUMEN
Most agronomic traits such as yield, quality and stress-tolerance of crops are quantitative traits. It is not easy to dissect genetic basis of these traits because they are controlled by multi-genes that are affected by environmental factors. Hitchhiking mapping offers a new method for identifying of loci controlling those traits and assessing their allelic variations. Marker-trait association analysis based on the hitchhiking effects will play an important role in dissection of complex traits. Combination of quantitative trait loci (QTL) mapping with marker-trait association analysis will facilitate dissection of complex traits, resulting in important information and markers for molecular breeding by design in crops. In this review, we presented a brief introduction of hitchhiking effects, marker-trait association analysis and practical considerations.
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Mapeo Cromosómico/métodos , Sitios de Carácter Cuantitativo/genética , Modelos GenéticosRESUMEN
Ubiquitin-mediated proteolysis is involved in many biological processes in eukaryotes. SCF complex is a very important ubiquitin E3 ligase which has been exploited very well in plants. F-box protein characterized by an F-box motif is a subunit of SCF complex, which works as determinant in substrate recognition. Currently, many F-box proteins have been identified in plants which are involving in hormone (e.g., ethylene, auxin, gibberellin and jasmonate ) signal transduction and biological processes, such as self-incompatibility and floral development. F-box proteins may also participate stress response in plants. Recent study suggested that the Arabidopsis F-box protein TIR1 is an auxin receptor. Therefore, F-box protein mediated proteolysis may be an important gene expression mechanism in plants.
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Proteínas F-Box/metabolismo , Desarrollo de la Planta , Proteínas de Plantas/metabolismo , Plantas/metabolismo , Proteínas F-Box/química , Fenómenos Fisiológicos de las Plantas , Proteínas de Plantas/química , Plantas/genética , Transducción de SeñalRESUMEN
OBJECTIVE: To study the expressions and activities of Rho GTPases in hypoxia and its relationship with tumor angiogenesis. METHODS: Three tumor cell lines were used in this study: gastric cancer cell lines AGS, SGC7901 and hepatocellular carcinoma cell line HepG2. Expression level of Rac1 mRNA was detected by semi-quantitative RT-PCR. Activity of Rac1 was determined by pull-down assay and expression of HIF-1alpha, VEGF, p53 and PTEN protein was detected by Westernblot. RESULTS: The expression level of Rac1 mRNA was significantly increased in hypoxia compared to normoxia. Pull-down assay showed that hypoxia-induced activity of Rac1 was elevated in a time-dependent manner and climaxed at 3 hours. The expressions of HIF-1alpha and VEGF protein were up-regulated, while those of PTEN and p53 protein were down-regulated. CONCLUSION: These results indicate that hypoxia enhances Rac1 expression which might be involved in tumor angiogenesis by reacting with hypoxia-responsive genes.