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Find video protocols related to scientific articles indexed in Pubmed.
Characterization of Protein Kinase PsSRPKL, a Novel Pathogenicity Factor in the Wheat Stripe Rust Fungus.
Environ. Microbiol.
PUBLISHED: 11-10-2014
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As in other eukaryotes, protein kinases (PKs) are generally evolutionarily conserved and play major regulatory roles in plant pathogenic fungi. Many PKs have been proven to be important for pathogenesis in model fungal plant pathogens, but little is currently known about their roles in the pathogenesis of cereal rust fungi, devastating pathogens in agriculture worldwide. Here, we report on an in planta highly induced PK gene PsSRPKL from the wheat stripe rust fungus Puccinia striiformis f. sp. tritici (Pst), one of the most important cereal rust fungi. PsSRPKL belongs to a group of PKs that are evolutionarily specific to cereal rust fungi. It shows a high level of intra-species polymorphism in the kinase domains and directed GFP-chimers to plant nuclei. Overexpression of PsSRPKL in fission yeast induces aberrant cell morphology and a decreased resistance to environmental stresses. Most importantly, PsSRPKL is proven to be an important pathogenicity factor responsible for fungal growth and responses to environmental stresses, therefore contributing significantly to Pst virulence in wheat. We hypothesize that cereal rust fungi have developed specific protein kinases as pathogenicity factors for adaptation to their host species during evolution. Thus our findings provide significant insights into pathogenicity and virulence evolution in cereal rust fungi.
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Cloning and characterization of AabHLH1, a bHLH transcription factor that positively regulates artemisinin biosynthesis in Artemisia annua.
Plant Cell Physiol.
PUBLISHED: 06-26-2014
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Amorpha-4,11-diene synthase (ADS) and Cyt P450 monooxygenase (CYP71AV1) in Artemisia annua L. are two key enzymes involved in the biosynthesis of artemisinin. The promoters of ADS and CYP71AV1 contain E-box elements, which are putative binding sites for basic helix-loop-helix (bHLH) transcription factors. This study successfully isolated a bHLH transcription factor gene from A. annua, designated as AabHLH1, from a cDNA library of the glandular secretory trichomes (GSTs) in which artemisinin is synthesized and sequestered. AabHLH1 encodes a protein of 650 amino acids containing one putative bHLH domain. AabHLH1 and ADS genes were strongly induced by ABA and the fungal elicitor, chitosan. The transient expression analysis of the AabHLH1-green fluorescent protein (GFP) reporter gene revealed that AabHLH1 was targeted to nuclei. Biochemical analysis demonstrated that the AabHLH1 protein was capable of binding to the E-box cis-elements, present in both ADS and CYP71AV1 promoters, and possessed transactivation activity in yeast. In addition, transient co-transformation of AabHLH1 and CYP71AV1Pro::GUS in A. annua leaves showed a significant activation of the expression of the GUS (?-glucuronidase) gene in transformed A. annua, but mutation of the E-boxes resulted in abolition of activation, suggesting that the E-box is important for the CYP71AV1 promoter activity. Furthermore, transient expression of AabHLH1 in A. annua leaves increased transcript levels of the genes involved in artemisinin biosynthesis, such as ADS, CYP71AV1 and HMGR. These results suggest that AabHLH1 can positively regulate the biosynthesis of artemisinin.
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Wheat TaNPSN SNARE homologues are involved in vesicle-mediated resistance to stripe rust (Puccinia striiformis f. sp. tritici).
J. Exp. Bot.
PUBLISHED: 06-24-2014
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Subcellular localisation of SNAREs (soluble N-ethylmaleimide-sensitive factor attachment protein receptors) and their ability to form SNARE complexes are critical for determining the specificity of vesicle fusion. NPSN11, a Novel Plant SNARE (NPSN) gene, has been reported to be involved in the delivery of cell wall precursors to the newly formed cell plate during cytokinesis. However, functions of NPSN genes in plant-pathogen interactions are largely unknown. In this study, we cloned and characterized three NPSN genes (TaNPSN11, TaNPSN12, and TaNPSN13) and three plant defence-related SNARE homologues (TaSYP132, TaSNAP34, and TaMEMB12). TaSYP132 showed a highly specific interaction with TaNPSN11 in both yeast two-hybrid and bimolecular fluorescence complementation (BiFC) assays. We hypothesize that this interaction may indicate a partnership in vesicle trafficking. Expressions of the three TaNPSNs and TaSYP132 were differentially induced in wheat leaves when challenged by Puccinia striiformis f. sp. tritici (Pst). In virus-induced gene silencing (VIGS) assays, resistance of wheat (Triticum aestivum) cultivar Xingzi9104 to the Pst avirulent race CYR23 was reduced by knocking down TaNPSN11, TaNPSN13 and TaSYP132, but not TaNPSN12, implying diversified functions of these wheat SNARE homologues in prevention of Pst infection and hyphal elongation. Immuno-localization results showed that TaNPSN11 or its structural homologues were mainly distributed in vesicle structures near cell membrane toward Pst hypha. Taken together, our data suggests a role of TaNPSN11 in vesicle-mediated resistance to stripe rust.
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Multiple genital tract tumors and mucinous adenocarcinoma of colon in a woman with Peutz-Jeghers syndrome: a case report and review of literatures.
Int J Clin Exp Pathol
PUBLISHED: 06-15-2014
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We report a very rare case of Peutz-Jeghers syndrome (PJS) composed of multiple genital tract tumors and mucinous adenocarcinoma. A 46-year-old woman presented to our hospital with lower abdominal pain resulting from PJS involves sex cord tumor with annular tubules (SCTAT), ovarian mucinous tumor, ovarian serous tumor, mucinous adenocarcinoma of colon. The CEA concentration is high before surgery, and decreases after the surgery and subsequent chemoradiotherapy. This case demonstrates a classic clinical presentation of a patient with PJS. PJS patients have increased risk of malignancy and early detection and regular surveillance of the high-risk patients with PJS is crucial. Surgery may be required for obstructive gastrointestinal lesions as well as those exhibiting malignant degeneration.
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Re-characterization of an extrachromosomal circular plasmid in the pathogenic Leptospira interrogans serovar Lai strain 56601.
Acta Biochim. Biophys. Sin. (Shanghai)
PUBLISHED: 05-29-2014
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In China, Leptospira interrogans serovar Lai strain 56601 (str.56601) is one of main pathogenic strains that cause severe leptospirosis in both human and animals. The genome of this organism was completely sequenced in 2003. However, in 2011, we identified and corrected some assembly errors in the str.56601 genome due to the repeat sequences widely distributed in the Leptospira genome. In this study, we re-analyzed the previously reported mobile, phage-related genomic island in the chromosome and rectified detailed sequence information in both the plasmid and chromosome using various experimental methods. The presence of a separate circular extrachromosomal plasmid was also confirmed, and its location in the genomic region was determined relative to the genomic island reported in L. interrogans serovar Lai by a combination of pulsed-field gel electrophoresis -based and plasmid extraction-based Southern blot analysis. This report confirmed that the separate extrachromosomal circular plasmid is not integrated into the chromosome of L. interrogans str.56601 and markedly improved our understanding of the genomic organization, evolution, and pathogenesis of L. interrogans. In particular, characterization of this extrachromosomal circular plasmid will contribute to the development of genetic manipulation systems in pathogenic Leptospira species.
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Subcellular localization of calcium in the incompatible and compatible interactions of wheat and Puccinia striiformis f. sp. tritici.
Protoplasma
PUBLISHED: 05-19-2014
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Ca(2+) is an ubiquitous intracellular molecule which is used as a second messenger to control many physiological activities in plant cells. In the present work, the relationship between calcium localization and the hypersensitive response (HR)-one of the most crucial and indispensable pathway to resist a pathogen-was studied in the wheat-wheat strip rust system using cytochemical technique. Our results show that calcium is involved in the interaction between wheat and wheat stripe rust. In the incompatible interaction associated with necrosis of host mesophyll cells, an influx of Ca(2+) from the intercellular space to the cytoplasm and finally an efflux to the intercellular space again was detected in an incompatible interaction. Calcium precipitates were also observed in mesophyll cells adjacent to necrotic cells. On the contrary, calcium flow was not significantly altered in a compatible interaction. These results suggest that calcium might induce HR as a secondary messenger in the incompatible interaction of wheat and wheat stripe rust.
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[A meta analysis of the effect of enhanced hand hygiene on the morbidity of ventilator-associated pneumonia].
Zhonghua Wei Zhong Bing Ji Jiu Yi Xue
PUBLISHED: 05-10-2014
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To determine the effect of enhanced hand hygiene on the morbidity of ventilator-associated pneumonia (VAP).
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[Health survey of painting and coating workers in an automobile manufacturing enterprise in Guangzhou, China].
Zhonghua Lao Dong Wei Sheng Zhi Ye Bing Za Zhi
PUBLISHED: 04-24-2014
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To investigate the health status of painting and coating workers in an automobile manufacturing enterprise in Guangzhou, China and analyze the influential factors for the health status of these workers, and to provide health intervention measures and strategies.
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Transcriptome profiling to identify genes involved in pathogenicity of Valsa mali on apple tree.
Fungal Genet. Biol.
PUBLISHED: 04-02-2014
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Apple Valsa canker, caused by the fungus Valsa mali (Vm), is one of the most destructive diseases of apple in China. A better understanding of this host-pathogen interaction is urgently needed to improve management strategies. In the current study we sequenced the transcriptomes of Vm during infection of apple bark and mycelium grown in axenic culture using Illumina RNA-Seq technology. We identified 437 genes that were differentially expressed during fungal infection compared to fungal mycelium grown in axenic culture. One hundred and thirty nine of these 437 genes showed more than two fold higher transcript abundance during infection. GO and KEGG enrichment analyses of the up-regulated genes suggest prevalence of genes associated with pectin catabolic, hydrolase activity and secondary metabolite biosynthesis during fungal infection. Some of the up-regulated genes associated with loss of pathogenicity and reduced virulence annotated by host-pathogen interaction databases may also be involved in cell wall hydrolysis and secondary metabolite transport, including a glycoside hydrolase family 28 protein, a peptidase and two major facilitator superfamily proteins. This highlights the importance of secondary metabolites and cell wall hydrolases during establishment of apple Valsa canker. Functional verification of the genes involved in pathogenicity of Vm will allow us to better understand how the fungus interferes with the host machinery and assists in apple canker establishment.
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Effect of high-fructose corn syrup on Streptococcus mutans virulence gene expression and on tooth demineralization.
Eur. J. Oral Sci.
PUBLISHED: 03-24-2014
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High-fructose corn syrup-55 (HFCS-55) has been widely welcomed in recent years as a substitute for sucrose on the basis of its favourable properties and price. The objective of this study was to determine the influence of HFCS-55 on the expression of Streptococcus mutans UA159 virulence genes and on tooth demineralization. Real-time reverse-transcription PCR (real-time RT-PCR) and microhardness evaluations were performed to examine gene expression and enamel demineralization, respectively, after treatment with HFCS-55 and/or sucrose. Significant up-regulation of glucosyltransferase B (gtfB) by HFCS-55 was found. A mixture of HFCS-55 and sucrose could positively enhance expression of glucan-binding protein (gbp) genes. Regarding acidogenicity, expression of the lactate dehydrogenase (ldh) gene was unaffected by HFCS-55. A notable finding in this study was that 5% HFCS-55 significantly enhanced expression of the intracellular response gene of the two-component VicRK signal transduction system (vicR). Demineralization testing showed that the microhardness of teeth decreased by a greater extent in response to HFCS-55 than in response to sucrose. The results indicate that HFCS-55 can enhance S. mutans biofilm formation indirectly in the presence of sucrose and that HFCS-55 has a more acidogenic potential than does sucrose. Summing up the real-time PCR and demineralization results, HFCS-55 appears to be no less cariogenic than sucrose in vitro - at least, not under the conditions of our experiments.
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Contrast-enhanced sonography of thyroid nodules.
J Clin Ultrasound
PUBLISHED: 03-21-2014
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To investigate the value of contrast-enhanced ultrasound (CEUS) in the differentiation of benign and malignant nodules.
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Agrobacterium tumefaciens-mediated transformation of the causative agent of Valsa canker of apple tree Valsa mali var. mali.
Curr. Microbiol.
PUBLISHED: 02-20-2014
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Valsa mali var. mali (Vmm), which is the causative agent of Valsa canker of apple tree, causes heavy damage to apple production in eastern Asia. In this article, we report Agrobacterium tumefaciens-mediated transformation (ATMT) of Vmm and expression of gfp (green fluorescent protein) in this fungus. The transformation system was optimized to a transformation efficiency of approximately 150 transformants/10(6) conidia, and a library containing over 4,000 transformants was generated. The tested transformants were mitotically stable. One hundred percent hph (hygromycin B phosphotransferase) integration into Vmm was identified by PCR and five single-copy integration of T-DNA was detected in the eighteen transformants by Southern blot. To our knowledge, this is the first report of ATMT of Vmm. Furthermore, this library has been used to identify genes involved in the virulence of the pathogen, and the transformation system may also be useful to the transformation of other species of the genus Valsa.
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Characterization of primary symptoms leading to Chinese patients presenting at hospital with suspected obstructive sleep apnea.
J Thorac Dis
PUBLISHED: 02-18-2014
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We identified the primary symptoms leading to Chinese patients presenting at hospital with suspected obstructive sleep apnea (OSA) and studied the prevalence and characteristics of OSA in confirmed cases.
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Regulation network and expression profiles of Epstein-Barr virus-encoded microRNAs and their potential target host genes in nasopharyngeal carcinomas.
Sci China Life Sci
PUBLISHED: 02-16-2014
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Epstein-Barr virus (EBV) is associated with nasopharyngeal carcinoma (NPC) tumorigenesis. However, the mechanism(s) connecting EBV infection and NPC remain unclear. Recently, a new class of EBV microRNAs (miRNAs) has been described. To determine how EBV miRNAs control the expression of host genes, and to understand their potential role in NPC tumorigenesis, we profiled the expression of 44 mature EBV miRNAs and potential host genes in NPC and non-tumor nasopharyngeal epithelial tissues. We found that 40 EBV miRNAs from the BART transcript were highly expressed in NPC. Analysis of potential BART miRNA target genes revealed that 3140 genes and several important pathways might be involved in the carcinogenesis of NPC. A total of 105 genes with potential EBV miRNA binding sites were significantly downregulated, suggesting that EBV miRNAs may regulate these genes and contribute to NPC carcinogenesis. An EBV miRNA and host gene regulation network was generated to provide useful clues for validating of EBV miRNA functions in NPC tumorigenesis.
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Predictive value of androgens and multivariate model for poor ovarian response.
Reprod. Biomed. Online
PUBLISHED: 02-11-2014
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No single or multivariate model is effective for predicting poor ovarian response (POR) with satisfactory sensitivity and specificity. This study investigated whether dehydroepiandrosterone sulphate (DHEAS) or basal testosterone concentrations could be effective predictors of POR defined by the Bologna criteria. This retrospective study included 79 poor responders and 128 normal responders. Serum FSH, LH, oestradiol, DHEAS and testosterone concentrations on day 3 of the menstrual cycle before the treatment cycle were measured. All patients received standard ovarian stimulation with FSH under pituitary suppression with gonadotrophin-releasing hormone agonist. DHEAS concentration was not significantly different between poor and normal responders or between pregnant and nonpregnant women. Basal testosterone, unlike DHEAS concentration, was predictive, but with limited ability as a single predictor, for POR. The multivariate model composed of age, AFC, FSH, FSH/LH and testosterone was reliably predictive for POR (ROC(AUC) = 0.976, cut-off point >0.51, sensitivity 88.6%, specificity 98.3%) and clinical pregnancy (ROC(AUC) = 0.716, cut-off point ?-0.22, sensitivity 75%, specificity 62.5%) and was better than antral follicle count for predicting both POR and clinical pregnancy. This multivariate model might be useful for identifying patients at risk of poor response in order to optimize the stimulation regimens. No single or multivariate model is effective for predicting poor ovarian response (POR) with satisfactory sensitivity and specificity. It has been suggested that androgens stimulate folliculogenesis and their concentrations might be correlated with oocyte yield after ovulation induction. We investigated whether dehydroepiandrosterone sulphate (DHEAS) or basal testosterone concentrations could be effective predictors for POR defined by the Bologna criteria. This is a retrospective study with 79 poor responders and 128 normal responders. Serum FSH, LH, oestradiol, DHEAS and testosterone on day 3 of the menstrual cycle before the treatment cycle were measured. All patients received standard ovarian stimulation with FSH under pituitary suppression with gonadotrophin-releasing hormone agonist. DHEAS concentration was not significantly different between poor and normal responders or between pregnant and nonpregnant women. Basal testosterone, instead of DHEAS, was predictive, but with limited ability as a single predictor, for POR. However, the multivariate model of (0.563 × Z(age)) ? (0.505 × Z(AFC)) + (0.506 × Z(FSH)) + (0.34 × Z(FSH/LH)) ? (0.24 × Z(testosterone)) (Z(Xi) = standardized values of variables) was reliably predictive for POR (ROC(AUC) = 0.976) and pregnancy (ROC(AUC) = 0.716) and was better than antral follicle count for predicting both POR and pregnancy. This multivariate model composed of age, AFC, FSH, FSH/LH and testosterone might be useful for identifying patients at risk of poor response in order to optimize the stimulation regimens.
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Construction and characterization of a bacterial artificial chromosome library for the hexaploid wheat line 92R137.
Biomed Res Int
PUBLISHED: 01-28-2014
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For map-based cloning of genes conferring important traits in the hexaploid wheat line 92R137, a bacterial artificial chromosome (BAC) library, including two sublibraries, was constructed using the genomic DNA of 92R137 digested with restriction enzymes HindIII and BamHI. The BAC library was composed of total 765,696 clones, of which 390,144 were from the HindIII digestion and 375,552 from the BamHI digestion. Through pulsed-field gel electrophoresis (PFGE) analysis of 453 clones randomly selected from the HindIII sublibrary and 573 clones from the BamHI sublibrary, the average insert sizes were estimated as 129 and 113 kb, respectively. Thus, the HindIII sublibrary was estimated to have a 3.01-fold coverage and the BamHI sublibrary a 2.53-fold coverage based on the estimated hexaploid wheat genome size of 16,700 Mb. The 765,696 clones were arrayed in 1,994 384-well plates. All clones were also arranged into plate pools and further arranged into 5-dimensional (5D) pools. The probability of identifying a clone corresponding to any wheat DNA sequence (such as gene Yr26 for stripe rust resistance) from the library was estimated to be more than 99.6%. Through polymerase chain reaction screening the 5D pools with Xwe173, a marker tightly linked to Yr26, six BAC clones were successfully obtained. These results demonstrate that the BAC library is a valuable genomic resource for positional cloning of Yr26 and other genes of interest.
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TaMDHAR4, a monodehydroascorbate reductase gene participates in the interactions between wheat and Puccinia striiformis f. sp. tritici.
Plant Physiol. Biochem.
PUBLISHED: 01-23-2014
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Reactive oxygen species (ROS) in plants are induced in various cellular compartments upon pathogen infection and act as an early signal during plant-pathogen interactions. Monodehydroascorbate reductase (MDHAR) is involved in plant disease resistance through the regulation of the ROS level via the ascorbate-glutathione (AsA-GSH) cycle. In this study, TaMDHAR4 was firstly isolated from wheat cultivar Suwon 11, and this protein exhibits high similarity to MDHAR proteins from other plant species. Bioinformatics analyses indicated that TaMDHAR4 contains typical structural features, such as mPTS-like sequences in the C-terminal extension and trans-membrane domain followed by five basic arginine residues (-RKRRR), which predicted that this protein may be localized in the peroxisome. qRT-PCR analyses demonstrated that TaMDHAR4 could be induced by various exogenous hormones, such as ABA, MeJA, and ETH. TaMDHAR4 is sharply down-regulated at 12 and 18 hpi only in wheat leaves challenged with Puccinia striiformis f. sp. tritici (Pst) race CYR23 and induced at 48 hpi with both Pst races CYR23 and CYR31. SOD and APX injection analyses demonstrated that TaMDHAR4 may be involved in the interaction between wheat and Pst through the regulation of its expression. Moreover, the knockdown of TaMDHAR4 through virus-induced gene silencing (VIGS) enhanced the wheat resistance to Pst by inhibiting sporulation in the compatible interaction. Histological observations also demonstrated that silenced wheat resulted in an increased proportion of necrotic area at the infection sites and suppressed Pst hypha elongation. The study provided novel insights into the molecular functions of TaMDHAR4 during plant-pathogen interactions.
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Delimiting cryptic pathogen species causing apple Valsa canker with multilocus data.
Ecol Evol
PUBLISHED: 01-09-2014
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Fungal diseases are posing tremendous threats to global economy and food safety. Among them, Valsa canker, caused by fungi of Valsa and their Cytospora anamorphs, has been a serious threat to fruit and forest trees and is one of the most destructive diseases of apple in East Asia, particularly. Accurate and robust delimitation of pathogen species is not only essential for the development of effective disease control programs, but also will advance our understanding of the emergence of plant diseases. However, species delimitation is especially difficult in Valsa because of the high variability of morphological traits and in many cases the lack of the teleomorph. In this study, we delimitated species boundary for pathogens causing apple Valsa canker with a multifaceted approach. Based on three independent loci, the internal transcribed spacer (ITS), ?-tubulin (Btu), and translation elongation factor-1 alpha (EF1?), we inferred gene trees with both maximum likelihood and Bayesian methods, estimated species tree with Bayesian multispecies coalescent approaches, and validated species tree with Bayesian species delimitation. Through divergence time estimation and ancestral host reconstruction, we tested the possible underlying mechanisms for fungal speciation and host-range change. Our results proved that two varieties of the former morphological species V. mali represented two distinct species, V. mali and V. pyri, which diverged about 5 million years ago, much later than the divergence of their preferred hosts, excluding a scenario of fungi-host co-speciation. The marked different thermal preferences and contrasting pathogenicity in cross-inoculation suggest ecological divergences between the two species. Apple was the most likely ancestral host for both V. mali and V. pyri. Host-range expansion led to the occurrence of V. pyri on both pear and apple. Our results also represent an example in which ITS data might underestimate species diversity.
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A Novel Fungal Hyperparasite of Puccinia striiformis f. sp. tritici, the Causal Agent of Wheat Stripe Rust.
PLoS ONE
PUBLISHED: 01-01-2014
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Puccinia striiformis f. sp. tritici (Pst), the causal fungus of wheat stripe rust, was previously reported to be infected by Lecanicillium lecanii, Microdochium nivale and Typhula idahoensis. Here, we report a novel hyperparasite on Pst. This hyperparasitic fungus was identified as Cladosporium cladosporioides (Fresen.) GA de Vries based on morphological characteristics observed by light and scanning electron microscopy together with molecular data. The hyperparasite reduced the production and viability of urediniospores and, therefore, could potentially be used for biological control of wheat stripe rust.
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Telomerase activity is more significant for predicting the outcome of IVF treatment than telomere length in granulosa cells.
Reproduction
PUBLISHED: 01-01-2014
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Our previous study has demonstrated that luteinized granulosa cells (GCs) have the potential to proliferate and that the telomerase activity (TA) of luteinized GCs may predict the clinical outcomes of IVF treatment. However, in the field of telomere research, there have always been different opinions regarding the significance of TA and telomere length (TL). Thus, in the present study, we compared the effects of these two parameters on IVF treatment outcomes in the same individuals. TL did not differ significantly between the pregnant group and the non-pregnant group. The TA, number of retrieved oocytes and rate of blastocyst transfer were significantly higher in the pregnant group than in the non-pregnant group (0.8825 OD×mm, 12.75±2.20 and 34.48%, respectively, in the pregnant group vs 0.513 OD×mm, 11.60±0.93 and 14.89%, respectively, in the non-pregnant group (P<0.05)), while basal FSH level was lower in the pregnant group than in the non-pregnant group. The subjects did not differ with regard to ovarian stimulation or other clinical characteristics. A TA increase of 1 OD×mm increased the chance of becoming pregnant 4.769-fold (odds ratio: 5.769, 95% CI: 1.434-23.212, P<0.014). The areas under the receiver operating characteristic curves were 0.576 for TL and 0.674 for TA (P=0.271 and P<0. 012 respectively). The corresponding cut-off points were 4.470 for TL and 0.650 OD×mm for TA. These results demonstrate that TA is a better predictor of pregnancy outcomes following IVF treatment than TL. No other clinical parameters, including age, baseline FSH level or peak oestradiol level, distinguished between the pregnant group and the non-pregnant group as effectively as TA.
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Endometriosis fertility index score maybe more accurate for predicting the outcomes of in vitro fertilisation than r-AFS classification in women with endometriosis.
Reprod. Biol. Endocrinol.
PUBLISHED: 08-31-2013
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Endometriosis is a common disease. The most widely used staging system of endometriosis is the revised American Fertility Society classification (r-AFS classification) which has limited predictive ability for pregnancy after surgery. The endometriosis fertility index (EFI) is used to predict fecundity after endometriosis surgery. This diagnostic accuracy study was designed to compare the predictive value of the EFI with that of the r-AFS classification for IVF outcomes in patients with endometriosis.
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Monodehydroascorbate reductase gene, regulated by the wheat PN-2013 miRNA, contributes to adult wheat plant resistance to stripe rust through ROS metabolism.
Biochim. Biophys. Acta
PUBLISHED: 08-14-2013
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Wheat stripe rust, caused by Puccinia striiformis f. sp. tritici (Pst), is one of the most destructive wheat diseases worldwide. Varieties with adult plant resistance (APR) maintain effective and durable disease resistance. APR to stripe rust in wheat cultivar XZ9104 (XZ) is associated with extensive hypersensitive cell death and production of reactive oxygen species in the host. MDHAR is an important gene in the AsA-GSH cycle, and it plays an important role in maintaining the reduced pool of AsA scavenging hydrogen peroxide. microRNAs (miRNAs) were shown to engage in post-transcriptional regulation by degrading target mRNAs or repressing gene translation in plants responding to abiotic/biotic stresses. Previously, two novel miRNAs (1136-P3 and PN-2013) were isolated in wheat and the target gene of them was determined using degradome sequencing technology. In this study, the target gene was isolated and characterized as TaMDHAR, a monodehydroascorbate reductase gene. We first demonstrated that the target gene could be cleaved by these two miRNAs in tobacco leaves experimentally. However, TaMDHAR was regulated by PN-2013, not 1136-P3, in wheat-Pst adult incompatible interaction according to the expression patterns. The TaMDHAR knockdown resulted in improved wheat resistance to Pst at the seedling stage, with no influence on 1136-P3 and PN-2013 expression. The TaMDHAR knockdown resulted in a much greater H2O2 accumulation and lower APX and CAT activities together with higher expression in several PR genes. We deduced that TaMDHAR could contribute to the APR of XZ through ROS metabolism as regulated by the AsA-GSH cycle.
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Functions of the lethal leaf-spot 1 gene in wheat cell death and disease tolerance to Puccinia striiformis.
J. Exp. Bot.
PUBLISHED: 07-02-2013
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Pheophorbide a oxygenase (PaO) is a key enzyme in chlorophyll catabolism that is known to suppress cell death in maize and Arabidopsis. The catalytic activity of PaO in chlorophyll degradation has been clearly demonstrated, but the function of PaO in the regulation of cell death and plant-microbe interactions is largely unknown. In this study, we characterized a PaO homologue in wheat of the lethal leaf-spot 1 gene, TaLls1, that was induced in leaves infected by Puccinia striiformis f.sp. tritici (Pst) and wounding treatment. The TaLls1 protein contains a conserved Rieske [2Fe-2S] motif and a mononuclear iron-binding site typical of PaOs. Silencing of TaLls1 by virus-induced gene silencing in wheat led to leaf cell death without pathogen attacks, possibly due to the accumulation of pheophorbide a (upstream substrate of PaO), indicating a suppressor role of TaLls1, while overexpression of TaLls1 also triggered cell death in both tobacco and wheat leaves, probably owing to the accumulation of the red chlorophyll catabolite (downstream product of PaO). Further deletion mutant analysis showed that the conserved Rieske domain, but not the iron-binding site, was essential for cell death induction. These results thus suggest a threshold for TaLls1 in maintaining cell homeostasis to adapt in various stresses, and shed new light on the role of TaLls1 in cell death regulation. Furthermore, silencing of TaLls1 in wheat did not change the disease symptoms but enhanced tolerance to Pst via an significant increase in H2O2 generation, elevated cell death occurrence, and upregulation of pathogenesis-related genes.
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Simultaneous osseous metaplasia nodules of the submucosa and mesosalpinx after first trimester abortion: a case report.
Eur. J. Med. Res.
PUBLISHED: 06-12-2013
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Here, we report a case of simultaneous osseous metaplasia nodules of the submucosa and mesosalpinx after a first trimester abortion.
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Wheat zinc finger protein TaLSD1, a negative regulator of programmed cell death, is involved in wheat resistance against stripe rust fungus.
Plant Physiol. Biochem.
PUBLISHED: 06-12-2013
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Genetic characterization of the Arabidopsis lesion simulating disease 1 (lsd1) mutant, a lesion mimic mutant (LMM), has revealed the essential role of AtLSD1 in the negative regulation of cell death and disease resistance. The three zinc-finger motifs found in AtLSD1 revealed a novel plant-specific gene family, whose members are significantly related to programmed cell death (PCD). In this study, we characterized a functional homologue to AtLSD1, TaLSD1, in the wheat-stripe rust fungus pathosystem. The expression of TaLSD1 was differentially induced during incompatible and compatible interactions between wheat and Puccinia striiformis f. sp. tritici (Pst) and was up-regulated by oxidative stress generated by methyl viologen (MV). TaLSD1 was found to be predominately localized in the nucleus of onion epidermal cell. Transient overexpression assays in Nicotiana benthamiana demonstrated that TaLSD1 partially inhibited programmed cell death triggered by a mouse Bax protein, whereas expression of TaLSD1 alone had no influence on the phenotype of tobacco. Knocking down the expression of TaLSD1 through virus-induced gene silencing (VIGS) increased wheat resistance against Pst accompanied by an enhanced hypersensitive response (HR), an increase in PR1 gene expression and a reduction in Pst hyphal growth. Our results suggest that TaLSD1 functions negatively in regulating the plant hypersensitive cell death and is involved in disease resistance of wheat against the stripe rust pathogen.
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TaEIL1, a wheat homologue of AtEIN3, acts as a negative regulator in the wheat-stripe rust fungus interaction.
Mol. Plant Pathol.
PUBLISHED: 06-03-2013
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Transcription factors (TFs) play crucial roles in the transcriptional regulation of plant development and defence responses. Increasing evidence has implicated ETHYLENE INSENSITIVE3 (EIN3) in the plant defence response to pathogen infection and environmental stimuli. However, the role of EIN3 in wheat resistance to Puccinia striiformis f. sp. tritici (Pst) is not clear. Here, TaEIL1 was isolated by rapid amplification of cDNA ends (RACE) based on a sequence fragment from a wheat-Pst interaction cDNA library. The TaEIL1 protein contains a typical EIN3-binding domain, and transient expression analyses indicated that TaEIL1 is localized in the nucleus. Yeast one-hybrid assay revealed that TaEIL1 exhibits transcriptional activity, and its C-terminus is necessary for the activation of transcription. TaEIL1 transcripts were regulated by environmental stress stimuli and were decreased under salicylic acid (SA) treatment. When wheat leaves were challenged with Pst, the transcript level of TaEIL1 in the compatible interaction was approximately three times higher than that in the incompatible interaction. Knocking down TaEIL1 through the Barley stripe mosaic virus (BSMV) virus-induced gene silencing (VIGS) system attenuated the growth of Pst, with shortened hyphae and reduced hyphal branches, haustorial mother cells and colony size. Moreover, enhanced necrosis was triggered by the Pst avirulent race CYR23, indicating that the hypersensitive response was strengthened in TaEIL1-silenced wheat plants. Thus, the up-regulation of defence-related genes and increased sucrose abundance might contribute to the enhanced disease resistance of wheat to the virulent race CYR31. Taken together, the results suggested that the suppression of TaEIL1 transcripts could enhance the resistance of wheat to stripe rust fungus.
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Elevated expression levels of matrix metalloproteinase-9 in placental villi and tissue inhibitor of metalloproteinase-2 in decidua are associated with prolonged bleeding after mifepristone-misoprostol medical abortion.
Fertil. Steril.
PUBLISHED: 05-30-2013
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To determine whether the expression levels of matrix metalloproteinases 2 and 9 (MMP-2 and -9) and tissue inhibitors of metalloproteinases 1 and 2 (TIMP-1 and -2) in the villi and the decidua are associated with prolonged bleeding after medical abortion.
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Hmgb1-TLR4-IL-23-IL-17A axis promote ischemia-reperfusion injury in a cardiac transplantation model.
Transplantation
PUBLISHED: 05-17-2013
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Cardiac transplantation is the last resort for patients with end-stage heart failure. Ischemia-reperfusion (IR) injury is a major issue in cardiac transplantation. Inflammatory processes play a major role in myocardial IR injury. However, the cellular and molecular immune mechanisms of myocardial IR injury remain elusive.
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Target of tae-miR408, a chemocyanin-like protein gene (TaCLP1), plays positive roles in wheat response to high-salinity, heavy cupric stress and stripe rust.
Plant Mol. Biol.
PUBLISHED: 05-03-2013
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microRNAs (miRNAs) are novel and significant regulators of gene expression at the post-transcriptional level, and they are essential for normal growth and development and adaptation to stress conditions. As miRNAs are a kind of RNAs that do not code proteins, they play roles by repressing gene translation or degrading the corresponding target mRNAs. Plantacyanin-like (basic blue) proteins have been predicted and verified as the target gene of miR408 in wheat and Arabidopsis, respectively. Besides some biochemical characteristics, their detailed biological function remains unknown. In this study, the target gene of a wheat miRNA (tae-miR408), designated TaCLP1, was identified using degradome sequencing and co-transformation technology in tobacco leaves. We isolated the full-length cDNA clone, and defined its product as a chemocyanin-like protein, a kind of plantacyanin. Transcript accumulation of TaCLP1 and tae-miR408 showed contrasting divergent expression patterns in wheat response to Puccinia striiformis f. sp. tritici (Pst) and high copper ion stress. Overexpression of TaCLP1 in yeast (Schizosaccharomyces pombe) significantly increased cell growth under high salinity and Cu²? stresses. Silencing of individual cDNA clones in wheat challenged with Pst indicated that TaCLP1 positively regulates resistance to stripe rust. The results indicate that the target of tae-miR408, TaCLP1, play an important role in regulating resistance of host plants to abiotic stresses and stripe rust, and such interactions can be a valuable resource for investigating stress tolerance in wheat.
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Long pentraxin PTX3 attenuates ischemia reperfusion injury in a cardiac transplantation model.
Transpl. Int.
PUBLISHED: 04-16-2013
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Ischemia reperfusion (IR) injury is a major issue in cardiac transplantation, and inflammatory processes play a major role in myocardial IR injury. Long pentraxin-3 (PTX3) is a member of a phylogenetically conserved group of acute-phase reactants that are involved in inflammation and innate immunity. In our study, hearts of C57Bl/6 mice were flushed and stored in cold Bretschneider solution for 8 h and then transplanted into syngeneic recipient. We found that both mRNA and protein levels of PTX3 were increased following myocardial IR injury; neutralizing antibody against PTX3 aggravated cardiomyocyte apoptosis and recruitment of neutrophils and macrophages. Troponin T (TnT) production on 24 h after myocardial IR injury was reduced by exogenous PTX3 administration and increased by PTX3 neutralization in comparison with control. Cardiac output at 60 mmHg of afterload pressure was also increased in hearts with exogenous PTX3 administration and decreased with PTX3 neutralization (PTX3: 58.4 ± 7.4 ml/min; Control: 24.5 ± 3.8 ml/min; Anti-PTX3: 11.6 ± 1.7 ml/min; P < 0.05). Furthermore, PTX3 restricted expansion of ?? T cell that was the major source of IL-17A and down-regulated expression of IL-23 and IL-17A. In conclusion, PTX3 played a protective role in cardiomyocyte IR injury. PTX3 ameliorated cardiomyocyte apoptosis and infiltration of neutrophil and macrophage and then improved hemodynamic performance. This was associated with restricted ?? T-cell expansion and decreased IL-23/IL-17A expression.
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Heavy metal pollution status in surface sediments of Swan Lake lagoon and Rongcheng Bay in the northern Yellow Sea.
Chemosphere
PUBLISHED: 04-06-2013
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The national Shandong Peninsula Blue Economic Zone Development Plan compels the further understanding of the distribution and potential risk of metals pollution in the east coast of China, where the rapid economic and urban development have been taken off and metal pollution has become a noticeable problem. Surface sediments collected from the largest swan habitat in Asia, the Swan Lake lagoon and the surrounding coastal area in Rongcheng Bay in northern Yellow Sea, were analyzed for the total metal concentrations and chemical phase partitioning of five heavy metals (Cu, Zn, Pb, Cd, and Cr). Metal contents in the studied region have increased significantly in the past decade. The speciation analyzed by the sequential extraction showed that Zn and Cr were present dominantly in the residual fraction and thus of low bioavailability, while Cd, Pb and Cu were found mostly in the non-residual fraction thus of high potential availability, indicating significant anthropogenic sources. Among the five metals, Cd is the most outstanding pollutant and presents high risk, and half of the surface sediments in the studied region had a 21% probability of toxicity based on the mean Effect Range-Median Quotient. At some stations with comparable total metal contents, remarkably different non-residual fraction portions were determined, pointing out that site-specific risk assessment integrating speciation is crucial for better management practices of coastal sediments.
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High genome heterozygosity and endemic genetic recombination in the wheat stripe rust fungus.
Nat Commun
PUBLISHED: 03-27-2013
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Stripe rust, caused by Puccinia striiformis f. sp. tritici (Pst), is one of the most destructive diseases of wheat. Here we report a 110-Mb draft sequence of Pst isolate CY32, obtained using a fosmid-to-fosmid strategy, to better understand its race evolution and pathogenesis. The Pst genome is highly heterozygous and contains 25,288 protein-coding genes. Compared with non-obligate fungal pathogens, Pst has a more diverse gene composition and more genes encoding secreted proteins. Re-sequencing analysis indicates significant genetic variation among six isolates collected from different continents. Approximately 35% of SNPs are in the coding sequence regions, and half of them are non-synonymous. High genetic diversity in Pst suggests that sexual reproduction has an important role in the origin of different regional races. Our results show the effectiveness of the fosmid-to-fosmid strategy for sequencing dikaryotic genomes and the feasibility of genome analysis to understand race evolution in Pst and other obligate pathogens.
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Clinical and polysomnographic features of patients with multiple system atrophy in Southwest China.
Sleep Breath
PUBLISHED: 03-22-2013
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The association between clinical features and polysomnographic variables of Chinese patients with multiple-system atrophy (MSA) remains largely unknown.
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Identification of eighteen Berberis species as alternate hosts of Puccinia striiformis f. sp. tritici and virulence variation in the pathogen isolates from natural infection of barberry plants in China.
Phytopathology
PUBLISHED: 03-22-2013
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ABSTRACT The wheat stripe rust pathogen (Puccinia striiformis f. sp. tritici) population in China has been reported to be a distinct genetic group with higher diversity than those in many other countries. Genetic recombination in the P. striiformis f. sp. tritici population has been identified with molecular markers but whether sexual reproduction occurs in China is unknown. In this study, we surveyed barberry plants for infection by rust fungi in the stripe rust "hotspot" regions in Gansu, Sichuan, and Shaanxi provinces; collected barberry plants and inoculated plants of 20 Berberis spp. with germinated teliospores under controlled greenhouse conditions for susceptibility to P. striiformis f. sp. tritici; and tested P. striiformis f. sp. tritici isolates obtained from aecia on naturally infected barberry plants on the wheat genotypes used to differentiate Chinese P. striiformis f. sp. tritici races to determine virulence variations. Different Berberis spp. were widely distributed and most surveyed plants had pycnia and aecia of rust fungi throughout the surveyed regions. In total, 28 Berberis spp. were identified during our study. From 20 Berberis spp. tested with teliospores of P. striiformis f. sp. tritici from wheat plants, 18 species were susceptible under greenhouse conditions. Among 3,703 aecia sampled from barberry plants of three species (Berberis shensiana, B. brachypoda, and B. soulieana) under natural infections in Gansu and Shaanxi provinces, four produced P. striiformis f. sp. tritici uredinia on susceptible wheat Mingxian 169. Sequence of the internal transcribed spacer (ITS) regions of the four isolates from barberry shared 99% identity with the P. striiformis f. sp. tritici sequences in the National Center for Biotechnology Information database. The four isolates had virulence patterns different from all previously reported races collected from wheat plants. Furthermore, 82 single-uredinium isolates obtained from the four barberry isolates had high virulence diversity rates of 9.0 to 28.1%, indicating that the diverse isolates were produced through sexual reproduction on barberry plants under natural conditions. In addition to P. striiformis f. sp. tritici, sequence analysis of polymerase chain reaction products of the ITS regions and inoculation tests on wheat identified P. graminis (the stem rust pathogen). Our results indicated that P. striiformis f. sp. tritici can infect some Berberis spp. under natural conditions, and the sexual cycle of the fungus may contribute to the diversity of P. striiformis f. sp. tritici in China.
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Validation of reference genes for gene expression analysis in Valsa mali var. mali using real-time quantitative PCR.
World J. Microbiol. Biotechnol.
PUBLISHED: 03-12-2013
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Valsa mali var. mali (Vmm), is the predominant species of apple valsa canker in China. Modern analysis of genes involved in virulence or pathogenicity usually implicate gene expression analysis most often performed using real-time quantitative polymerase chain reaction (RT-qPCR). However, for relative gene expression analysis pertinent reference genes have to be validated before using them as internal reference. This has not been reported for Vmm, so far. Therefore, eight commonly used housekeeping genes (ACT, CYP, EF1-?, G6PDH, GAPDH, L13, TUB, and UBQ) were cloned and evaluated for their expression stability by geNorm and NormFinder. Overall, all of the candidate reference genes were found to be suitable for gene expression analysis. After analysis of 10 samples from different strains and abiotic stress treatments, G6PDH appeared to be the most suitable reference gene, whereas GAPDH was the least suitable. Moreover, taking G6PDH combined with L13 or CYP as reference genes, improved the reliability of RT-qPCR significantly. The influence of the reference system on expression data was demonstrated by analyzing Vmmpg-1 encoding an endo-polygalacturonase gene. Pectinases are considered key pathogenicity factors for this fungus. In order to better understand the role of pectinases in pathogenicity of Vmm, RT-qPCR was used for expression analysis. Our results may provide a guideline for future studies on gene expression of V. mali var. mali by using RT-qPCR.
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Polysomnographically determined sleep and body mass index in patients with insomnia.
Psychiatry Res
PUBLISHED: 02-09-2013
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We assessed associations between polysomnographically determined sleep, especially the amount of slow-wave sleep (SWS), and body mass index (BMI) in patients with insomnia. One hundred and forty-one insomniacs and 55 healthy volunteers completed overnight polysomnographic recordings, and we measured height and body weight. No significant correlations were obtained between total sleep time and BMI among insomniacs. Compared with normal volunteers, insomnia patients exhibited longer sleep latency and shorter total sleep duration. While the two groups had no significant differences in BMI, insomniacs presented with more N1 but less time spend in SWS and rapid eye movement sleep (REMS). Based on their SWS time, we divided insomnia patients into three groups: short (26.99±13.88), intermediate (59.24±8.12), and long (102.21±26.17) SWS groups. The short-SWS group had significantly greater BMI than the long-SWS group. Further analyses with multiple linear regression showed a significant negative correlation between the amount of SWS and BMI scores in insomniacs, whereas no such correlation was found in healthy volunteers after controlling for potential confounds (e.g., age, sex and AHI). Our study suggests that low amounts of SWS may be associated with higher BMI in patients with insomnia.
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Fine mapping of wheat stripe rust resistance gene Yr26 based on collinearity of wheat with Brachypodium distachyon and rice.
PLoS ONE
PUBLISHED: 01-27-2013
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The Yr26 gene, conferring resistance to all currently important races of Puccinia striiformis f. sp. tritici (Pst) in China, was previously mapped to wheat chromosome deletion bin C-1BL-6-0.32 with low-density markers. In this study, collinearity of wheat to Brachypodium distachyon and rice was used to develop markers to saturate the chromosomal region containing the Yr26 locus, and a total of 2,341 F2 plants and 551 F2?3 progenies derived from Avocet S×92R137 were used to develop a fine map of Yr26. Wheat expressed sequence tags (ESTs) located in deletion bin C-1BL-6-0.32 were used to develop sequence tagged site (STS) markers. The EST-STS markers flanking Yr26 were used to identify collinear regions of the rice and B. distachyon genomes. Wheat ESTs with significant similarities in the two collinear regions were selected to develop conserved markers for fine mapping of Yr26. Thirty-one markers were mapped to the Yr26 region, and six of them cosegregated with the resistance gene. Marker orders were highly conserved between rice and B. distachyon, but some rearrangements were observed between rice and wheat. Two flanking markers (CON-4 and CON-12) further narrowed the genomic region containing Yr26 to a 1.92 Mb region in B. distachyon chromosome 3 and a 1.17 Mb region in rice chromosome 10, and two putative resistance gene analogs were identified in the collinear region of B. distachyon. The markers developed in this study provide a potential target site for further map-based cloning of Yr26 and should be useful in marker assisted selection for pyramiding the gene with other resistance genes.
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Equilibrious strand exchange promoted by DNA conformational switching.
Sci Rep
PUBLISHED: 01-24-2013
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Most of DNA strand exchange reactions in vitro are based on toehold strategy which is generally nonequilibrium, and intracellular strand exchange mediated by proteins shows little sequence specificity. Herein, a new strand exchange promoted by equilibrious DNA conformational switching is verified. Duplexes containing c-myc sequence which is potentially converted into G-quadruplex are designed in this strategy. The dynamic equilibrium between duplex and G4-DNA is response to the specific exchange of homologous single-stranded DNA (ssDNA). The SER is enzyme free and sequence specific. No ATP is needed and the displaced ssDNAs are identical to the homologous ssDNAs. The SER products and exchange kenetics are analyzed by PAGE and the RecA mediated SER is performed as the contrast. This SER is a new feature of G4-DNAs and a novel strategy to utilize the dynamic equilibrium of DNA conformations.
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Associations of acanthosis nigricans with metabolic abnormalities in polycystic ovary syndrome women with normal body mass index.
J. Dermatol.
PUBLISHED: 01-05-2013
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Acanthosis nigricans (AN) usually correlates to insulin resistance (IR) or obesity in obese populations, but adequate studies on the significance of AN in people with normal body mass index (BMI) have not been performed and discussed. Three hundred and thirty-nine polycystic ovary syndrome (PCOS) patients with normal BMI (<23 kg/m(2) ) were recruited. The anthropometric and biochemical parameters of these patients were measured. In these patients with normal BMI, 33 (9.7%) women had AN, and six (1.77%) women were diagnosed with metabolic syndrome. Most of the anthropometric and biochemical variables associated with metabolic status were more unfavorable in the AN-positive group compared with the AN-negative groups. The prevalence of central obesity, IR and reduced high-density lipoprotein cholesterol (HDL-C) level were also significantly higher in the AN-positive group (P < 0.05). In multiple regression analysis, presence of AN was still significantly associated with IR (odds ratio [OR] = 2.952, 95% confidence intervals [CI] = 1.367-6.376] and reduced HDL-C level (OR = 2.668, 95% CI = 1.160-6.135) after adjustments for age and BMI. Sensitivity, specificity, and positive and negative predictive values for AN to detect IR were 18.6%, 92.6%, 39.4% and 81.4%, respectively. In conclusion, presence of AN correlated with IR and reduced HDL-C level in PCOS women with normal BMI. AN status had high specificity to detect IR, but lack of sensitivity.
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Fractal analysis of permeability of unsaturated fractured rocks.
ScientificWorldJournal
PUBLISHED: 01-01-2013
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A physical conceptual model for water retention in fractured rocks is derived while taking into account the effect of pore size distribution and tortuosity of capillaries. The formula of calculating relative hydraulic conductivity of fractured rock is given based on fractal theory. It is an issue to choose an appropriate capillary pressure-saturation curve in the research of unsaturated fractured mass. The geometric pattern of the fracture bulk is described based on the fractal distribution of tortuosity. The resulting water content expression is then used to estimate the unsaturated hydraulic conductivity of the fractured medium based on the well-known model of Burdine. It is found that for large enough ranges of fracture apertures the new constitutive model converges to the empirical Brooks-Corey model.
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[Experimental study on anti-metastasis effect of emodin on human pancreatic cancer].
Zhongguo Zhong Yao Za Zhi
PUBLISHED: 12-21-2011
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To investigate the anti-metastasis effect of emodin on the pancreatic cancer in vitro and in vivo.
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[Development of genetic transformation system of Valsa mali of apple mediated by PEG].
Wei Sheng Wu Xue Bao
PUBLISHED: 12-01-2011
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The genetic transformation of Valsa mali var. mali was developed by PEG-mediated protoplasts transformation.
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Endogenous hydrogen peroxide is a key factor in the yeast extract-induced activation of biphenyl biosynthesis in cell cultures of Sorbus aucuparia.
Planta
PUBLISHED: 09-12-2011
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Biphenyls are unique phytoalexins produced by plants belonging to Pyrinae, a subtribe of the economically important Rosaceae family. The formation of aucuparin, a well-known biphenyl, is induced by yeast extract (YE) in cell cultures of Sorbus aucuparia. However, the molecular mechanism underlying YE-induced activation of biphenyl biosynthesis remains unknown. Here we demonstrate that the addition of YE to the cell cultures results in a burst of reactive oxygen species (ROS; H(2)O(2) and O(2) (-)), followed by transcriptional activation of the biphenyl synthase 1 gene (BIS1) encoding the key enzyme of the biphenyl biosynthetic pathway and aucuparin accumulation. Pretreatment of the cell cultures with ROS scavenger dihydrolipoic acid and NADPH oxidase-specific inhibitor diphenylene iodonium abolished all of the above YE-induced biological events. However, when the cell cultures was pretreated with superoxide dismutase specific inhibitor N,N-diethyldithiocarbamic acid, although O(2) (-) continued to be generated, the H(2)O(2) accumulation, BIS1 expression and aucuparin production were blocked. Interestingly, exogenous supply of H(2)O(2) in the range of 0.05-10 mM failed to induce aucuparin accumulation. These results indicate that endogenous generation of H(2)O(2) rather than that of O(2) (-) is a key factor in YE-induced accumulation of biphenyl phytoalexins in cell cultures of S. aucuparia.
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Human neonatal neutrophils are resistant to apoptosis with lower caspase-3 activity.
Tohoku J. Exp. Med.
PUBLISHED: 08-27-2011
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The incidence of neonatal inflammatory diseases remains high despite improved strategies for dealing with infection. Neutrophils are believed to play a significant role in neonatal inflammatory diseases due to their secretion of harmful mediators. Neutrophils rapidly undergo apoptosis following activation; dysregulation of the neutrophil apoptotic pathway may be an underlying mechanism of neonatal inflammatory disorders. In this study, we determined whether neonatal neutrophils are intrinsically resistant to apoptosis relative to their adult counterparts. Twelve healthy full-term newborn infants and 12 healthy adult volunteers, aged 20 to 45 years, were enrolled in this study. Neutrophils were isolated from umbilical cord blood or fresh venous peripheral blood, and neutrophils of each subject were cultured for up to 48 hours. Flow cytometric analysis revealed that spontaneous apoptosis of adult neutrophils increased with culture time (23% ± 2% at 12 h, 49% ± 4% at 24 h and 76% ± 3% at 48 h), whereas the frequency of apoptosis was significantly lower in neutrophils from neonates (9% ± 2% at 12 h, 30% ± 4% at 24 h and 49% ± 3% at 48 h) (p < 0.01 for each time point). Importantly, the expression levels of caspase-3 mRNA and a precursor form of caspase-3 protein were lower in neonatal neutrophils than adult neutrophils, as judged by RT-PCR and Western blot analyses. Moreover, caspase-3 activity was lower in neonatal neutrophils, compared to adult neutrophils. These findings suggest that neonatal neutrophils are intrinsically apoptosis-resistant, which may be due to the low expression of caspase-3.
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Long-range pseudoknot interactions dictate the regulatory response in the tetrahydrofolate riboswitch.
Proc. Natl. Acad. Sci. U.S.A.
PUBLISHED: 08-22-2011
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Tetrahydrofolate (THF), a biologically active form of the vitamin folate (B(9)), is an essential cofactor in one-carbon transfer reactions. In bacteria, expression of folate-related genes is controlled by feedback modulation in response to specific binding of THF and related compounds to a riboswitch. Here, we present the X-ray structures of the THF-sensing domain from the Eubacterium siraeum riboswitch in the ligand-bound and unbound states. The structure reveals an "inverted" three-way junctional architecture, most unusual for riboswitches, with the junction located far from the regulatory helix P1 and not directly participating in helix P1 formation. Instead, the three-way junction, stabilized by binding to the ligand, aligns the riboswitch stems for long-range tertiary pseudoknot interactions that contribute to the organization of helix P1 and therefore stipulate the regulatory response of the riboswitch. The pterin moiety of the ligand docks in a semiopen pocket adjacent to the junction, where it forms specific hydrogen bonds with two moderately conserved pyrimidines. The aminobenzoate moiety stacks on a guanine base, whereas the glutamate moiety does not appear to make strong interactions with the RNA. In contrast to other riboswitches, these findings demonstrate that the THF riboswitch uses a limited number of available determinants for ligand recognition. Given that modern antibiotics target folate metabolism, the THF riboswitch structure provides insights on mechanistic aspects of riboswitch function and may help in manipulating THF levels in pathogenic bacteria.
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Microscopy and proteomic analysis of the non-host resistance of Oryza sativa to the wheat leaf rust fungus, Puccinia triticina f. sp. tritici.
Plant Cell Rep.
PUBLISHED: 08-11-2011
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Rice (Oryza sativa) cv. Nipponbare expresses non-host resistance (NHR) to the wheat leaf rust fungus, Puccinia triticina f. sp. tritici (Ptt). When the leaves of cv. Nipponbare were inoculated with Ptt, approx 93% of the urediniospores germinated on the leaf surface, but only 10% of the germinated spores formed appressoria over the stomata at one day post inoculation (1 dpi). Hydrogen peroxide (H(2)O(2)) accumulated in host cells around the appressoria at 3 dpi. Approx. 3% of the appressoria produced short hyphae inside the leaf, and fluorescence was observed in tissue invaded by the hyphae by 7 dpi. At 22 dpi, 0.2% of the sites with appressoria formed branching infection hypha in mesophyll cells, but no substomatal vesicles, haustorial mother cells or haustoria were observed. Proteins were extracted from leaves 3 dpi and analyzed by two-dimensional gel electrophoresis (2-DE). A total 33 spots were reproducibly up-regulated and 9 were down-regulated by infection compared to the water inoculated control. Of these, 30 were identified by MALDI-TOF Mass Spectrometry. The identified proteins participate in defense/stress responses, energy/carbohydrate metabolism, oxidation-reduction processes, protein folding/turnover/cleavage/degradation, signal transduction and cell death regulation. The results indicates that NHR of rice to Ptt is consistent with a shift in protein and energy metabolism, increased antimicrobial activities, possibly including phytoalexin accumulation and cell wall reinforcement, increased cell repair, antioxidive and detoxification reactions, and enhanced prevention of plant cell death. Nearly half of the up-regulated identified proteins were associated with chloroplast and mitochondrial physiology suggesting important roles for these organelles during NHR.
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Beclin1/PI3K-mediated autophagy prevents hypoxia-induced apoptosis in EAhy926 cell line.
Cancer Biother. Radiopharm.
PUBLISHED: 06-28-2011
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Although hypoxia can induce cell death, the cancer cells and endothelial cells within a solid tumor that remain active in the hypoxia microenvironment often possess an enhanced survival potential. Developing approaches aimed at increasing the sensitivity of endothelial cells to hypoxia-induced cell death represents a potentially important avenue for antiangiogenesis treatment. This study investigated approaches to increase the sensitivity of endothelial cells to hypoxia-induced apoptosis. Autophagy and apoptosis of endothelial cells induced by hypoxia were investigated by transmission electron microscopy, confocallaser microscopy, and western blotting. Moreover, cell invasion was observed by a transwell assay and F-actin quantitative analysis. In this study, it was found that hypoxia could induce both autophagy and apoptosis in hypoxia-inducible factor-1- and Beclin1-dependent endothelial cells. Hypoxia-induced autophagy was prohibited by phosphatidylinositol 3-kinase/Akt inhibitor but not mitogen-activated protein kinase inhibitor. Inhibition of autophagy promoted the rate of apoptosis. Further, the reversal of hypoxia-induced autophagy increased cell migration compared with the normoxia condition. This study concludes that hypoxia triggers a feedback mechanism that delays apoptosis of endothelial cells and that is driven by hypoxia-induced autophagy. Thus, approaches aimed at the disruption of this mechanism can be expected to enhance the susceptibility of endothelial cells to hypoxia-induced apoptosis.
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Regulation of AMH and SCF expression in human granulosa cells by GnRH agonist and antagonist.
Pharmazie
PUBLISHED: 06-25-2011
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With the progress of cancer treatment, fertility preservation has become an urgent requisition. Gonadotropin-releasing hormone agonist (GnRH-a) and antagonist (GnRH-ant) have been used to protect fertility for these patients. However, studies showed that although GnRH-a and GnRH-ant had a comparable down-regulating effect on the pituitary, GnRH-ant could not preserve ovarian function. Moreover, GnRH-ant alone could deplete primordial follicles. It might be speculated that an additional intraovarian system existed except the pituitary system. Anti-Müerian hormone (AMH) and Stem cell factor (SCF) proved to be the key factors in follicle recruitment and development. The balance between AMH and SCF was tightly related to ovarian reserve. To investigate the intraovarian effect of GnRH-a or GnRH-ant on ovarian reserve, we examined AMH and SCF expression in human granulosa cells (hGCs). GCs were isolated from follicular aspirates after oocyte removal from the patients undergoing assisted reproduction techniques. After pretreated with GnRH-a (triptorelin) or GnRH-ant (cetrorelix) for 48 h, mRNA and protein of AMH and SCF were analyzed by Real-time PCR and Immunoblot assay respectively. The results indicated that AMH mRNA and protein expressions were down-regulated in the GnRH-ant groups, SCF mRNA and protein expressions were up-regulated in the high-dose GnRH-ant group. There was no difference of AMH and SCF expression in the GnRH-a group or GnRH-a + GnRH-ant group compared with control. These results suggested the effects of GnRH-a and GnRH-ant on the regulation of AMH and SCF were different, which may provide insight into the mechanism of GnRH-a and GnRH-ant interventions on ovarian reserve.
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Mifepristone may shorten the induction-to-abortion time for termination of second-trimester pregnancies by ethacridine lactate.
Contraception
PUBLISHED: 05-31-2011
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We reviewed our experience with adding mifepristone to the protocol for the termination of pregnancy up to 24 weeks of gestation by intra-amniotic ethacridine lactate.
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Molecular characterization of a Fus3/Kss1 type MAPK from Puccinia striiformis f. sp. tritici, PsMAPK1.
PLoS ONE
PUBLISHED: 04-03-2011
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Puccinia striiformis f. sp. tritici (Pst) is an obligate biotrophic fungus that causes the destructive wheat stripe rust disease worldwide. Due to the lack of reliable transformation and gene disruption method, knowledge about the function of Pst genes involved in pathogenesis is limited. Mitogen-activated protein kinase (MAPK) genes have been shown in a number of plant pathogenic fungi to play critical roles in regulating various infection processes. In the present study, we identified and characterized the first MAPK gene PsMAPK1 in Pst. Phylogenetic analysis indicated that PsMAPK1 is a YERK1 MAP kinase belonging to the Fus3/Kss1 class. Single nucleotide polymerphisms (SNPs) and insertion/deletion were detected in the coding region of PsMAPK1 among six Pst isolates. Real-time RT-PCR analyses revealed that PsMAPK1 expression was induced at early infection stages and peaked during haustorium formation. When expressed in Fusarium graminearum, PsMAPK1 partially rescued the map1 mutant in vegetative growth and pathogenicity. It also partially complemented the defects of the Magnaporthe oryzae pmk1 mutant in appressorium formation and plant infection. These results suggest that F. graminearum and M. oryzae can be used as surrogate systems for functional analysis of well-conserved Pst genes and PsMAPK1 may play a role in the regulation of plant penetration and infectious growth in Pst.
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Histological and molecular studies of the non-host interaction between wheat and Uromyces fabae.
Planta
PUBLISHED: 03-28-2011
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Non-host resistance (NHR) confers plant species immunity against the majority of microbes. As an important crop, wheat can be damaged by several Puccinia species but is immune to all Uromyces species. Here, we studied the basis of NHR in wheat against the broad bean rust pathogen Uromyces fabae (Uf). In the wheat-Uf interaction, microscopic observations showed that urediospores germinated efficiently on wheat leaves. However, over 98% of the germ tubes failed to form appressoria over stomata. For the few that invaded through stomata, the majority of them failed to penetrate wheat mesophyll cells. At 96 hours after inoculation, less than 4% of the Uf infection units that had entered the mesophyll tissue formed haustoria. Attempted penetration by haustorium mother cells induced the thickening of cell wall and the formation of papillae in plant cells, which arrested the development or growth of Uf penetration pegs. For the Uf haustoria formed in wheat cells, they were encased in callose-like materials and did not elicit hypersensitive response. Localized accumulation of H(2)O(2) were observed in plant cell walls, papillae and encasement of haustoria during the wheat-Uf interaction. Furthermore, quantitative RT-PCR analysis showed that several genes involved in basal resistance and oxidative stress responses were up-regulated during Uf infection. In conclusion, our study revealed the cytological and molecular bases of NHR in wheat against the non-adapted rust fungus Uf, and highlighted the significance of papilla production in the prehaustorial NHR.
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The impact of the catastrophic earthquake in Chinas Sichuan province on the mental health of pregnant women.
J Affect Disord
PUBLISHED: 03-10-2011
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On May 12, 2008, a magnitude 8.0 earthquake struck Chinas southwestern Sichuan province. Recent studies have identified mental health problems among the survivors, but little is known about the impact of the Sichuan earthquake on the mental health of pregnant women in the area. The main objective was to assess the impact of the Sichuan earthquake on the mental health of pregnant women in earthquake stricken areas.
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Development and validation of a liquid chromatography-tandem mass spectrometry method for the determination of zofenopril and its active metabolite zofenoprilat in human plasma.
J Pharm Biomed Anal
PUBLISHED: 02-09-2011
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A novel, sensitive and rapid liquid chromatographic-electrospray ionization mass spectrometric method was developed and validated for the determination of zofenopril and its active metabolite zofenoprilat in human plasma. The method was based on a single extraction step using methyl tert-butyl ether and did not require chemical derivatization. The chromatographic conditions were optimized; separation was performed on a phenyl-hexyl column (5?m, 250mm×4.6mm i.d.) with a mobile phase consisting of a solution of methanol and water (95:5, v/v) that also contained 0.1% of formic acid. A flow rate of 1.0mL/min was used. Zofenopril, zofenoprilat and the internal standard (IS) fosinopril sodium were measured using an electrospray ion source in a positive reaction monitoring mode. Linear calibration curves were generated for zofenopril concentrations between 0.1052 and 1052ng/mL and for zofenoprilat concentrations between 0.2508 and 2508ng/mL. In both cases, the coefficients of determination were greater than 0.995. The extraction recovery for zofenopril was 93.5% on average. It was 92.5% for zofenoprilat. The inter- and intra-batch precision and accuracy for both zofenopril and zofenoprilat were higher than 14%. The method was applied to measure the concentrations of zofenopril and zofenoprilat in plasma samples.
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Self-assembled highly ordered ethane-bridged periodic mesoporous organosilica and its application in HPLC.
J Sep Sci
PUBLISHED: 02-07-2011
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Monodisperse spherical periodic mesoporous organosilicas (PMOs) with ethane integrated in the framework were synthesized and their application as stationary phase for chromatographic separation is demonstrated. The ethane-PMOs were prepared by condensation of 1,2-bis(triethoxysilyl)ethane (BTSE) in basic condition using octadecyltrimethylammonium chloride (C(18)TMACl) as template and ethanol as co-solvent. The morphology and mesoporous structure of ethane-PMOs were controlled under different concentrations of sodium hydroxide (NaOH) and EtOH. The results of scanning electron microscopy (SEM), transmission electron microscopy (TEM), powder X-ray diffraction (XRD), nitrogen sorption measurement, Fourier transform infrared spectroscopy (FT-IR) and elemental analysis showed that ethane-PMOs have spherical morphology, uniform particle distribution, highly ordered pore structure, high surface area and narrow pore-size distribution. The column packed with these materials exhibits good permeability, high chemical stability and good selectivity of mixtures of aromatic hydrocarbons in normal phase high-performance liquid chromatography (HPLC).
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Effects of low-dose mifepristone administration in two different 14-day regimens on the menstrual cycle and endometrial development: a randomized controlled trial.
Contraception
PUBLISHED: 01-20-2011
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This study was conducted to evaluate the effects of treatment with mifepristone 5 mg given every day or every other day for 14 days, starting 15 days after menses started for endometrial development.
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Inhibitory action of Celastrol on hypoxia-mediated angiogenesis and metastasis via the HIF-1? pathway.
Int. J. Mol. Med.
PUBLISHED: 01-18-2011
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Celastrol, a natural biologically active compound isolated from Tripterygium wilfordii Hook F root extracts, has been shown to possess antitumor properties and therefore, is an interesting candidate for the development of novel chemotherapeutic cancer agents. In this study, we have demonstrated that Celastrol is a potent inhibitor of hypoxia-induced angiogenic and metastatic activity as shown by a decrease in the proliferation of both endothelial and cancer cells, blocking of migration as well as of tube formation of endothelial cells, and by inhibition of cancer cell invasion under hypoxic conditions. Moreover, Celastrol decreased hypoxia-inducible factor-1? (HIF-1?) mRNA levels under both normoxia and hypoxia and inhibited hypoxia-induced accumulation of nuclear HIF-1? protein. Meanwhile, inhibition of nuclear HIF-1? protein levels were accompanied by a reduction in the transcriptional activity of HIF-1? target genes, including VEGF. In addition, the inhibitory effect of Celastrol on HIF-1? protein was partly due to its suppression of HSP90 activity. We conclude that Celastrol regulates HIF-1? at multiple levels that may together or individually contribute to its antitumor activity against hypoxia-induced angiogenesis and metastasis.
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Effect of human endometrial stromal cell-derived conditioned medium on uterine natural killer (uNK) cells proliferation and cytotoxicity.
Am. J. Reprod. Immunol.
PUBLISHED: 01-12-2011
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Human endometrial stromal cells are involved in the regulation of immune cell proliferation, apoptosis, differentiation, and function. In the endometrium, uNK cells are in close contact with stromal cells. The aim of the study was to investigate the effects of human endometrial stromal cells on uNK-cell proliferation and uNK-cell cytotoxicity.
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Polysomnographic characteristics of daytime sleepiness in obstructive sleep apnea syndrome.
Sleep Breath
PUBLISHED: 01-08-2011
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The study is intended to explore the nocturnal sleep determinants for excessive daytime sleepiness (EDS) in patients with obstructive sleep apnea syndrome (OSAS).
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Prevalence and characterization of human Shigella infections in Henan Province, China, in 2006.
J. Clin. Microbiol.
PUBLISHED: 11-10-2010
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In 2006, 3,531 fecal samples were collected from patients with diarrhea in Henan Province, China. A total of 467 (13.2%) Shigella strains were isolated and serotyped. Seventy-one Shigella flexneri strains were characterized by MIC determination, pulsed-field gel electrophoresis (PFGE), and detection of genes encoding cephalosporin resistance. Most infections were caused by S. flexneri variant X [IV:(7),8] (27.6%), S. sonnei (24.2%), and S. flexneri 2a (20.8%). However, large regional differences were observed. Significantly higher odds (2.0) of females compared to males were infected with S. flexneri 2a. Untypeable S. flexneri (-:6) isolates were absent among males, as were untypeable S. flexneri [I:(7),8] isolates among females. Patient ages ranged from 2 months to 82 years, with 231 subjects (49.7%) <5 years of age. Most of the patients were male (62.1% [n = 290]). Infections peaked in July; week 27 with 38 cases (8.1%). All of the 71 S. flexneri conferred resistance to nalidixic acid; in addition, 21% (n = 15) and 79% (n = 56) were high- and low-level resistant to ciprofloxacin, respectively. Six S. flexneri isolates {serotype 2b [II:7,(8)] and 2b [II:(3),4;7,(8)]} harbored the bla(CTX-M-14) or bla(CTX-M-15) gene. A total of 52 unique XbaI PFGE patterns were observed among the 71 S. flexneri isolates with 11 distinct PFGE clusters. This study revealed a high prevalence of shigellosis with geographical differences in the distribution of serotypes in the distribution of serotypes and also differences in comparisons by gender. A high frequency of resistance, including 100% resistance to ciprofloxacin and resistance to extended-spectrum cephalosporins, was observed. We detected several isolates exhibiting the same PFGE type and MIC profile, indicating multiple undetected outbreaks.
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Re-evaluation of pathogens causing Valsa canker on apple in China.
Mycologia
PUBLISHED: 10-07-2010
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Valsa canker is a destructive disease on apple that causes serious economic losses in eastern Asia. In the present study fungal isolates from cankered apple and pear bark were examined and compared with morphology and rDNA-ITS sequences. Valsa mali was confirmed to be an independent species and a causal pathogen of Valsa canker on apple and pear in China. It was the predominant species (96.7% of isolates) on apple and was complemented by V. malicola (3.3% of isolates). Significant intraspecific genetic differentiation was detected in V. mali with two varieties recognized, V. mali var. mali occurring exclusively on apple and V. mali var. pyri occurring on both apple and pear. Results from genetic analysis and cross-inoculation tests provided support for the hypothesis that host preference probably catalyzed such genetic changes within the pathogen populations.
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[Cloning and transcriptional profiling of PsNCS1 from Puccinia striiformis f. sp. tritici].
Wei Sheng Wu Xue Bao
PUBLISHED: 09-07-2010
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We cloned PsNCS1 encoding neuronal calcium sensor from Pst and analyzed its transcriptional profile.
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Growth hormone releasing peptide-2, a ghrelin agonist, attenuates lipopolysaccharide-induced acute lung injury in rats.
Tohoku J. Exp. Med.
PUBLISHED: 09-01-2010
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Acute lung injury (ALI) and its severe form, acute respiratory distress syndrome (ARDS), are the most common complications of sepsis, and the mortality of sepsis-induced ALI remains high in critically ill patients. Growth hormone releasing peptide-2 (GHRP-2), a ghrelin agonist, has been shown to exert beneficial effects on various inflammatory diseases. We therefore explored whether GHRP-2 possesses anti-inflammatory properties in the pathogenesis of lipopolysaccharide (LPS)-induced ALI. Male Sprague-Dawley rats were intratracheally instilled with LPS (2 mg/kg) to induce ALI. ALI was confirmed with lung tissue injury (histopathological examination), enhanced lung edema (wet-to-dry weight ratio), and neutrophil infiltration (myeloperoxidase activity) at 6 h after LPS exposure. The analyses of bronchoalveolar lavage fluid showed the significant increases in pulmonary permeability (total cells and protein) and the levels of proinflammatory cytokines, including tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6). In contrast, these lung injury indexes were attenuated in rats that received a subcutaneous injection of GHRP-2 (100 microg/kg) 0.5 h prior to LPS administration. To further explore the potential anti-inflammatory mechanism of GHRP-2 in LPS-induced ALI, we assessed of nuclear factor-kappaB (NF-kappaB) activity in lung tissues at 6 h after LPS challenge. We thus found that pretreatment with GHRP-2 markedly suppressed the activation of NF-kappaB in lung tissues. These results indicate that GHRP-2 attenuated LPS-induced ALI. Early protection appears to be mediated partly through the inhibition of NF-kappaB pathway activation. The present study indicates that GHRP-2 acts as a potential therapeutic reagent for treating ALI.
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Metabolic abnormalities in adolescents with polycystic ovary syndrome in south China.
Reprod. Biol. Endocrinol.
PUBLISHED: 08-19-2010
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Adults with polycystic ovary syndrome (PCOS) can have multiple metabolic abnormalities. However, studies in the adolescent population are still limited and these results seem to vary widely. This study was to investigate the metabolic abnormalities in adolescents with PCOS in South China and the potential risk factors contributed to these health risks.
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Cloning and characterization of a calcium binding EF-hand protein gene TaCab1 from wheat and its expression in response to Puccinia striiformis f. sp. tritici and abiotic stresses.
Mol. Biol. Rep.
PUBLISHED: 05-09-2010
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Calcium is a ubiquitous and essential secondary messenger in eukaryotic signal transduction pathways. Calcium binding protein, as a component of pathways, plays various roles in response to biotic and abiotic stresses, as well as in developmental processes in plants. In this study, a calcium binding protein gene, designated as TaCab1 (Triticum aestivum calcium binding EF-hand protein 1), was isolated and characterized from wheat leaves (cv. Suwon 11) infected by Puccinia striiformis f. sp. tritici by in silico cloning and reverse transcription PCR (RT-PCR). TaCab1 did not have an intron and was predicted to encode a 216 amino acid protein which possesses an N-terminal region with a signal peptide, a transmembrane domain, an EF-hand motif and a caleosin domain. The results of transient assays with constructs of TaCab1 with green fluorescent protein (GFP) gene indicated that TaCab1 encodes a transmembrane protein. Quantitative real-time PCR (qRT-PCR) analyses revealed that TaCab1 was highly expressed in leaves than roots and stems. Although up-regulated expression profiles of TaCab1 were quite similar in both incompatible and compatible interactions, its transcript accumulation in the compatible interaction was much higher than in the incompatible interaction. The transcription of TaCab1 was also up-regulated at different degrees after treated by phytohormones [abscisic acid, benzyl adenine, ethylene, methyl jasmonate and salicylic acid (SA)] and stress stimuli [wounding, low temperature, polyethylene glycol and high salinity]. These results suggest that TaCab1 is involved in the plant-pathogen recognition, symptom development, and the basal tolerance to biotic and abiotic stresses through the SA signaling pathway.
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[Cloning and transcriptional profiling of a cell division cycle gene PsCdc2 from stripe rust fungus during incompatible and compatible interaction between wheat and Puccinia striiformis f. sp. tritici].
Wei Sheng Wu Xue Bao
PUBLISHED: 04-15-2010
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We cloned a cell division cycle PsCdc2 from Puccinia striiformis f. sp. tritici (Pst) and analyze its expression profile.
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Structural insights into ligand recognition by a sensing domain of the cooperative glycine riboswitch.
Mol. Cell
PUBLISHED: 04-12-2010
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Glycine riboswitches regulate gene expression by feedback modulation in response to cooperative binding to glycine. Here, we report on crystal structures of the second glycine-sensing domain from the Vibrio cholerae riboswitch in the ligand-bound and unbound states. This domain adopts a three-helical fold that centers on a three-way junction and accommodates glycine within a bulge-containing binding pocket above the junction. Glycine recognition is facilitated by a pair of bound Mg(2+) cations and governed by specific interactions and shape complementarity with the pocket. A conserved adenine extrudes from the binding pocket and intercalates into the junction implying that glycine binding in the context of the complete riboswitch could impact on gene expression by stabilizing the riboswitch junction and regulatory P1 helix. Analysis of riboswitch interactions in the crystal and footprinting experiments indicates that adjacent glycine-sensing modules of the riboswitch could form specific interdomain interactions, thereby potentially contributing to the cooperative response.
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Cloning and characterization of a wheat neutral ceramidase gene Ta-CDase.
Mol. Biol. Rep.
PUBLISHED: 03-29-2010
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Ceramidases are key enzymes in the regulation of the cellular levels of ceramide, sphingosine and sphingosine-1-phosphate. This study first reports on the molecular cloning, sequencing and expression profile of the gene encoding the wheat neutral ceramidase designated as Ta-CDase. A full length wheat Ta-CDase gene is obtained by rapid amplification of cDNA ends (RACE) based on the sequence of the WSRC36 fragment from an incompatible suppression subtractive hybridization (SSH) cDNA library of wheat leaves infected by Puccinia striiformis f. sp. tritici. The open reading frame (ORF) of 2,839 nucleotides encodes a polypeptide of 785 amino acids with a predicted isoelectric point (pI) of 6.398. The protein conserved domain search indicates that the polypeptide contains the signature of ceramidase, signal peptide sequence and transmembrane region. A phylogenetic analysis reveals that a high degree of relatedness exists among wheat Ta-CDase and ceramidases from other plant species at the amino acid level, while its relationship to that of animals and pathogens is more distant. The expression profile of the Ta-CDase shows a very strong expression of transcripts only at 48 h post inoculation (hpi), while expression level is low at other time points. Southern blot analyses showed that Ta-CDase is a multi-copy gene and located on wheat chromosome 4D and 5A.
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Characterization of the expression profile of a wheat aci-reductone-dioxygenase-like gene in response to stripe rust pathogen infection and abiotic stresses.
Plant Physiol. Biochem.
PUBLISHED: 03-03-2010
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The methionine salvage pathway is conserved from prokaryotes to high eukaryotes. The reaction catalyzed by aci-reductone-dioxygenase (ARD) represents a branch point in the methionine salvage pathway. A novel aci-reductone-dioxygenase gene, designed as TaARD, was identified in a subtraction library constructed with RNA isolated from wheat leaves infected with the stripe rust pathogen. TaARD was predicted to encode a 197 amino acid protein that belongs to the cupin superfamily. In transient expression assays with onion epidermal cells, the TaARD-GFP fusion protein localized to the nucleus and cytoplasm. Southern blot analysis showed that the wheat genome had multiple copies of TaARD. Quantitative real-time RT-PCR (qRT-PCR) analyses revealed that the TaARD transcript was induced in wheat leaves infected with a compatible stripe rust strain. However, its expression was reduced or suppressed in incompatible interactions and by ABA, ethephon (ET), or salicylic acid (SA) treatments. With methyl jasmonate (MeJA) treatment, TaARD transcript level was suppressed in the first 6h but increased afterwards. The expression of TaARD also was inhibited by wounding and environmental stimuli, including high salinity and low temperature. Because of the role of ARD in the methionine salvage pathway, these results suggest that TaARD may be involved in ethylene synthesis and ethylene signaling in response to biotic and abiotic stresses.
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Optimization of the fermentation process of actinomycete strain hhs.015.
J. Biomed. Biotechnol.
PUBLISHED: 02-28-2010
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Strain Hhs.015(T) (Saccharothrix yanglingensis sp. nov.), an antagonistic endophytic Saccharothrix actinomycete isolated from roots of cucumber plants, exhibited a broad antimicrobial spectrum in vitro and was active as a biocontrol against plant diseases in field trials. The SSY medium was used for production of antimicrobial metabolites by strain Hhs.015(T). However, this medium is too expensive for large-scale production. In this study, an alternative culture medium, based on agricultural waste products (e.g., apple pomace), was optimized. The results showed that the alternative medium contained 15 g apple pomace, 4 g rapeseed meal, 0.1 g KH(2)PO(4), and 0.6 g MgSO(4).7H(2)O in 1 L distilled water. This medium reduced the material costs by 91.5% compared to SSY medium. Response surface methodology (RSM) was used to investigate the influence of environmental variables on production of compounds of antimicrobial metabolites. The optimal conditions achieved were initial pH 7.0, medium volume of 90 mL in 250 mL flasks, rotary speed of 100 rpm, temperature 25 degrees C, and inoculation volume of 15.8%. The antimicrobial activity was increased by 20% by optimizing the environmental parameters. The results obtained allow an efficient production of components with antimicrobial activity by strain Hhs.015(T) on a large scale at low costs.
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