ABSTRACT Introduction: Severe fever with thrombocytopenia syndrome virus (SFTSV) is a newly emerged virus that poses a great threat to human health because of high fatality rate. Methods: To develop sensitive and specific sero-diagnostic systems for SFTSV infections, monoclonal antibodies (MAbs) against recombinant SFTSV nucleocapsid (rSFTSV-N) protein were developed by immunizing BALB/C mice with rSFTSV-N protein and fusing the spleen cells with SP2/0 myeloma cells. Three hybridoma cell lines secreting MAbs against rSFTSV-N were obtained. MAb based IgG sandwich enzyme linked immunosorbent assay (ELISA) and IgM capture ELISA systems were established by using the newly developed MAbs. One hundred fifteen clinical suspected SFTS patients serum samples were used to evaluate the newly established systems by comparing with the total antibody detecting sandwich ELISA system and indirect ELISA systems. Results: The MAbs based sandwich IgG ELISA was perfectly matched with that of the total antibody sandwich ELISA and the indirect IgG ELISA. IgM capture ELISA results perfectly matched with that of the total antibody sandwich ELISA while detecting eight additional positive samples missed by the indirect IgM ELISA. Conclusions: The MAbs against rSFTSV-N protein offer new tools for SFTSV studies and our newly developed MAb-based IgG and IgM capture ELISA systems would offer safe and useful tools for diagnosis of SFTS virus infections and epidemiological investigations.

OBJECTIVE: To determine the role of RNA-binding protein with serine-rich domain 1 (RNPS1) in uterine corpus endometrial carcinoma (UCEC), the role of RNPS1 knockdown in UCEC development in vitro and in vivo, and the relationship between RNPS1 and mismatch repair (MMR) in UCEC. METHODS: We predicted the potential function of RNPS1 using bioinformatics systems. The expression of RNPS1 in tissues and cell lines was analyzed by western blotting and immunohistochemistry. The expression of RNPS1 in MMR was assessed using bioinformatics and western blotting. The proliferation and apoptosis of UCEC cells were assessed under RNPS1 knockdown conditions, and RNPS1 regulation in MMR was detected by suppressing Notch signaling. Associations between RNPS1 and gene mutations in UCEC and prognosis were analyzed. RESULTS: The RNPS1 level was higher in UCEC tumors than in normal tissues and tumors or RL952 cells. Prognostic outcomes were worse when UCEC showed abundant RNPS1 expression. Lentiviral RNPS1 knockdown weakened tumor cell proliferation and suppressed biomarker expression, reduced the tumor volume, promoted apoptosis in vitro and in vivo, and inhibited UCEC development. Increased MutS homolog 2 (MSH2) and MutS homolog 6 (MSH6) levels in MMR after RNPS1 knockdown were reversed by inhibiting Notch signaling. Furthermore, RNPS1 was associated with mutations in NAA11, C2orf57, NUPR1, and other genes involved in UCEC prognosis. CONCLUSION: RNPS1 may regulate the expression levels of MSH2 and MSH6 in MMR, enhancing the proliferation, development, and prognosis of UCEC through a Notch signaling pathway in UCEC. Our study offers a new method and strategy for delaying UCEC development through modulating MMR.

Abstract Rapeseed is one of important oil crops in China. Better understanding of the regulation network of main agronomic traits of rapeseed could improve the yielding of rapeseed. In this study, we obtained an influrescence mutant that showed a fusion phenotype, similar with the Arabidopsis clavata-like phenotype, so we named the mutant as Bnclavata-like (Bnclv-like). Phenotype analysis illustrated that abnormal development of the inflorescence meristem (IM) led to the fused-inflorescence phenotype. At the stage of protein abundance, major regulators in metabolic processes, ROS metabolism, and cytoskeleton formation were seen to be altered in this mutant. These results not only revealed the relationship between biological processes and inflorescence meristem development, but also suggest bioengineering strategies for the improved breeding and production of Brassica napus.

Abstract Senescence, a highly programmed process, largely determines yield and quality of crops. However, knowledge about the onset and progression of leaf senescence in crop plants is still limited. Here, we report that salt-induced protein (salT), a new gene, may be involved in leaf senescence. Overexpressing salT could prolong the duration of leaves with higher concentrations of chlorophyll compared with the wild type. Moreover, overexpression of salT could delay the senescence of rice leaves though the inhibition of senescence associated genes (SAGs). Overall, the characterization of salT suggested that it is a new gene affecting the leaf senescence induced by natural and dark conditions.

Abstract The clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated (Cas) technology has recently emerged as a powerful genomic editing tool with great potential for crop breeding. However, commonly used protocols for screening of CRISPR/Cas9-induced mutations are laborious, time-consuming, and costly. In the present study we examined the applicability of high resolution melting (HRM) analysis fast screening CRISPR/Cas9-induced mutations in T0 plants and subsequent genotyping of T1 populations. Comparative analysis demonstrated that HRM analysis could identify mutant T0 plants carrying various types of mutation, including single nucleotide substitutions and short insertion/deletions, with false positive/negative rates of 0-2.78%. Furthermore, T1 plants derived from single T0 plants could be correctly genotyped by HRM analysis using WT parents and T0 plants as controls. We hence recommend the adoption of HRM analysis in CRISPR/Cas9 mediated genetic studies and breeding in rice and other crop species.

RESUMO: A salinidade do solo limita a produção agrícola. O trigo mole é uma das culturas mais importantes com característica allohexaploid e genoma altamente complexo. O mapeamento QTL é uma maneira muito útil de identificar genes para traços quantitativos, como a tolerância à salinidade em trigo hexaplóide. No presente estudo realizou-se um ensaio hidropónico para identificar locos de traços quantitativos (QTLs) associados à tolerância à salinidade do trigo sob concentração de NaCl 150 mM, usando uma população de linhagem consanguíneo recombinante (Xiaoyan 54 × Jing 411). Os valores dos traços de mudas de trigo, incluindo comprimento máximo da raiz (MRL), peso seco da raiz (RDW), ponha o peso seco (SDW), peso seco total (TDW) e a proporção das plantas de trigo TDW entre o estresse salgado e o controle (TDWR), foram avaliados ou calculados. Um total de 19QTLs para cinco traços foram detectados através do método de mapeamento de intervalo composto usando a versão 2.5 do cartógrafo QTL sob condições normais e de estresse salino. Estes QTLs distribuídos em 12 cromossomos explicaram a porcentagem de variação fenotípica por QTL individual variando de 7,9% a 19,0%. Entre eles, foram detectados 11 e 6 QTLs em condições de estresse normal e sal, respectivamente, e dois QTLs foram detectados para TDWR. Cromossoma 1A, 3A e 7A podem conter genes que são candidatos cruciais associados à tolerância ao sal de trigo. Nossos resultados seriam úteis para a seleção assistida por marcadores para produzir variedades de trigo com tolerância salina melhorada.

ABSTRACT: Soil salinity limits agricultural production and is a major obstacle for increasing crop yield. Common wheat is one of the most important crops with allohexaploid characteristic and a highly complex genome. QTL mapping is a useful way to identify genes for quantitative traits such as salinity tolerance in hexaploid wheat. In the present study, a hydroponic trial was carried out to identify quantitative trait loci (QTLs) associated with salinity tolerance of wheat under 150mM NaCl concentration using a recombinant inbred line population (Xiaoyan 54×Jing 411). Values of wheat seedling traits including maximum root length (MRL), root dry weight (RDW), shoot dry weight (SDW), total dry weight (TDW) and the ratio of TDW of wheat plants between salt stress and control (TDWR) were evaluated or calculated. A total of 19QTLs for five traits were detected through composite interval mapping method by using QTL Cartographer version 2.5 under normal and salt stress conditions. These QTLs distributed on 12 chromosomes explained the percentage of phenotypic variation by individual QTL varying from 7.9% to 19.0%. Among them, 11 and six QTLs were detected under normal and salt stress conditions, respectively and two QTLs were detected for TDWR. Some salt tolerance related loci may be pleiotropic. Chromosome 1A, 3A and 7A may harbor crucial candidate genes associated with wheat salt tolerance. Our results would be helpful for the marker assisted selection to breed wheat varieties with improved salt tolerance.

Abstract Background: CircRNAs are widespread in plants and play important roles in response to abiotic stresses. Low nitrogen (LN) promotes the growth of plant root system, allowing it to explore more nitrogen. However, whether circRNAs involved in the response to LN stress and the regulation of LN-promoted root growth in wheat remains unclear. Methods: Two wheat varieties (LH9 and XN979) with contrasting root phenotypes to LN stress were used as materials to identify circRNAs under control and LN conditions by using high-throughput sequencing technology. Results: Six differentially expressed circRNAs (DECs) involved in the common response to LN stress and 23 DECs involved in the regulation of LN-promoted root growth were successfully identified. GO analysis of the DEC-host genes involved in the regulation of LN-promoted root growth showed that GO terms related to biological regulation, responses to stimuli and signalling were significantly enriched. Moreover, seven DECs were predicted to have miRNA binding sites and may serve as miRNA sponges to capture miRNAs from their target genes. Conclusions: LN stress altered the expression profiles of circRNAs in wheat. This is the first report of LN stress responsive circRNAs in plants. Our results provided new clues for investigating the functions of circRNAs in response to LN stress and in the regulation of LN-promoted wheat root growth.

Neste trabalho, grafeno foi usado na extração em fase sólida dispersiva de 11 herbicidas triazina e 5 inseticidas neonicotina da água. As condições de extração como quantidade de grafeno, solvente de dessorção e pH da solução foram otimizados. Nas condições otimizadas, um enriquecimento de alta eficiência foi alcançado na análise quantitativa dos pesticidas. Triazinas e neonicotinas foram determinadas por espectrometria de massas-cromatografia gasosa (GC-MS) e cromatografia líquida de alta eficiência acoplada à espectrometria de massas (LC-MS/MS), respectivamente. Os resultados indicaram que o grafeno foi um excelente adsorvente na adsorção de pesticidas. A recuperação dos pesticidas foi 83,0-108,9% com desvio padrão relativo (RSD) entre 2,4% e 12,3%. Os limites de detecção (LODs) variaram entre 0,03 μg L-1 e 0,40 μg L-1. Finalmente, o método proposto foi aplicado na análise de amostras reais, como água de torneira e de rio.

In this work, graphene was used for dispersive solid-phase extraction of 11 triazine herbicides and 5 neonicotine insecticides from water. The extraction conditions such as the amount of graphene, the desorption solvent and the solution pH were optimized. Under the optimal conditions, high efficient enrichment was achieved for the quantitative analysis of the pesticides. Triazines and neonicotines were determined by gas chromatograph-mass spectrometer (GC-MS) and high performance liquid chromatography electrospray tandem mass spectrometry (LC-MS/MS), respectively. The results indicated that graphene was an excellent adsorbent for the adsorption of pesticides. The recoveries of the pesticides were 83.0-108.9% with relative standard deviation (RSD) between 2.4% and 12.3%. The limits of detection (LODs) ranged from 0.03 μg L-1 to 0.40 μg L-1. Finally, the proposed method was applied for real sample analysis, such as tap and river water.

Seeds are the most important plant storage organ and play a central role in the life cycle of plants. Since little is known about the protein composition of rice (Oryza sativa) seeds, in this work we used proteomic methods to obtain a reference map of rice seed proteins and identify important molecules. Overall, 480 reproducible protein spots were detected by two-dimensional electrophoresis on pH 4-7 gels and 302 proteins were identified by MALDI-TOF MS and database searches. Together, these proteins represented 252 gene products and were classified into 12 functional categories, most of which were involved in metabolic pathways. Database searches combined with hydropathy plots and gene ontology analysis showed that most rice seed proteins were hydrophilic and were related to binding, catalytic, cellular or metabolic processes. These results expand our knowledge of the rice proteome and improve our understanding of the cellular biology of rice seeds.

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OBJECTIF: Évaluer dans quelle mesure le peuple chinois est protégé contre les dépenses catastrophiques et l'appauvrissement dû aux dépenses médicales, mais aussi étudier le système de santé et les facteurs structurels qui influencent le premier de ces résultats. MÉTHODES: Des données ont été obtenues de la quatrième enquête nationale sur les services de santé. Une analyse des dépenses de santé catastrophiques et de l'appauvrissement dû aux dépenses médicales a été réalisée sur un échantillon de 55 556 ménages présentant diverses caractéristiques et vivant dans des environnements ruraux et urbains de différentes régions du pays. La régression logistique a été utilisée pour identifier les déterminants des dépenses de santé catastrophiques. RÉSULTATS: Le taux des dépenses de santé catastrophiques s'élevait à 13,0 % et celui de l'appauvrissement à 7,5%. Les taux de dépenses de santé catastrophiques étaient supérieurs dans les familles dont certains membres étaient hospitalisés, souffraient de maladies chroniques ou étaient des personnes âgées, ainsi que dans les familles des régions rurales ou plus pauvres. Une combinaison de facteurs défavorables augmentait le risque de dépenses de santé catastrophiques. Les familles bénéficiant de régimes d'assurance de résident ou d'employé urbain présentaient des taux de dépenses de santé catastrophiques inférieurs aux familles bénéficiant du nouveau régime corporatif rural. Le besoin en soins de santé et leur utilisation, les données démographiques, le type d'assurance et le type de méthode de paiement des fournisseurs étaient les déterminants des dépenses de santé catastrophiques. CONCLUSION: Bien que la Chine ait considérablement développé la couverture de son assurance-maladie, la protection financière reste insuffisante. L'objectif des responsables politiques doit être de créer des programmes d'assurance améliorés en étendant les prestations sociales, en redessinant les arrangements du partage des coûts et les méthodes de paiements des fournisseurs, mais aussi en développant des stratégies de contrôle des dépenses plus efficaces.

OBJECTIVE: To assess the degree to which the Chinese people are protected from catastrophic household expenditure and impoverishment from medical expenses and to explore the health system and structural factors influencing the first of these outcomes. METHODS: Data were derived from the Fourth National Health Service Survey. An analysis of catastrophic health expenditure and impoverishment from medical expenses was undertaken with a sample of 55 556 households of different characteristics and located in rural and urban settings in different parts of the country. Logistic regression was used to identify the determinants of catastrophic health expenditure. FINDINGS: The rate of catastrophic health expenditure was 13.0%; that of impoverishment was 7.5%. Rates of catastrophic health expenditure were higher among households having members who were hospitalized, elderly, or chronically ill, as well as in households in rural or poorer regions. A combination of adverse factors increased the risk of catastrophic health expenditure. Families enrolled in the urban employee or resident insurance schemes had lower rates of catastrophic health expenditure than those enrolled in the new rural corporative scheme. The need for and use of health care, demographics, type of benefit package and type of provider payment method were the determinants of catastrophic health expenditure. CONCLUSION: Although China has greatly expanded health insurance coverage, financial protection remains insufficient. Policy-makers should focus on designing improved insurance plans by expanding the benefit package, redesigning cost sharing arrangements and provider payment methods and developing more effective expenditure control strategies.

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OBJETIVO: Evaluar el grado de protección de los habitantes de China frente a los gastos familiares catastróficos y el empobrecimiento provocado por los gastos médicos y examinar el sistema sanitario y los factores estructurales que influyen en el primero de estos resultados. MÉTODOS: Los datos se obtuvieron de la 4ª Encuesta nacional sobre los servicios sanitarios. Se emprendió la tarea de analizar los gastos sanitarios catastróficos y el empobrecimiento provocado por los gastos médicos con una muestra de 55.556 hogares con características diferentes localizados tanto en entornos rurales como urbanos en distintas partes del país. Se empleó un modelo de regresión logística para identificar los determinantes del gasto sanitario catastrófico. RESULTADOS: La tasa del gasto sanitario catastrófico fue del 13.0% y la del empobrecimiento, del 7,5%. La tasa del gasto sanitario imprevisto fue superior en los hogares en los que alguno de los miembros estaba hospitalizado, era anciano o sufría una enfermedad crónica, así como en los hogares de zonas rurales o más pobres. Una combinación de factores adversos aumentó el riesgo de sufrir gastos sanitarios catastróficos. Las familias inscritas en los seguros urbanos para empleados o residentes presentaron una tasa menor de gastos sanitarios catastróficos que aquellas que estaban inscritas en el nuevo seguro corporativo rural. La necesidad y el uso de la atención sanitaria, la demografía, el tipo de prestaciones así como el método de pago al proveedor fueron los determinantes del gasto sanitario imprevisto. CONCLUSIÓN: Aunque China ha ampliado mucho la cobertura del seguro de salud, la protección financiera sigue siendo insuficiente. Las autoridades deberían centrarse en diseñar planes de seguros mejorados aumentando las prestaciones, rediseñando los acuerdos para la financiación de los gastos y los métodos de pago al proveedor y desarrollar estrategias más eficaces para el control de los gastos.

Desenvolveu-se um procedimento de microextração por emulsificação assistida por ultrassom e aperfeiçoada por surfactante (UASEME) combinado com cromatografia líquida de alta eficiência com detector de arranjo de diodos para a determinação de 17b-estradiol (βE2), estrona (E1) e dietilestilbestrol (DES) em água. Ultrassom foi aplicado para auxiliar a emulsificação e Triton X-100 foi usado como dispersor e emulsificante. Tetracloreto de carbono foi usado como solvente extrator. Empregando otimização global, o procedimento foi caracterizado por um intervalo linear aceitável de 10 a 1000 ng mL-1 para βE2, E1 e DES (r > 0,997), exatidão e precisão validadas (RSD 0,85-1,28% (n = 5)) e alta sensibilidade com limites de detecção de 0,200, 0,100 e 0,125 ng mL-1 para βE2, E1 e DES, respectivamente. O procedimento foi aplicado para a análise de amostras típicas de água, com fatores de enriquecimento de 85,29, 173,45 e 97,05 para βE2, E1 e DES, respectivamente, e boas recuperações (≥ 89,82%). De maneira geral, o procedimento desenvolvido foi simples e confiável, com potencial de aplicação em larga escala para análises da água.

An ultrasound-assisted surfactant-enhanced emulsification microextraction (UASEME) combined with high performance liquid chromatography-diode array detector procedure was developed for the determination of 17β-estradiol (βE2), estrone (E1) and diethylstilbestrol (DES) in water. Ultrasound was applied to assist emulsification, and Triton X-100 was adopted as the disperser and emulsifier. Carbon tetrachloride was applied as the extraction solvent. Global optimization was used and the developed procedure was characterized by an acceptable linear range of 10 to 1000 ng mL-1 for βE2, E1 and DES (r > 0.997), a validated accuracy and precision (RSD 0.85-1.28% (n = 5)), and high sensitivity with limits of detection of 0.200, 0.100 and 0.125 ng mL-1 for βE2, E1 and DES, respectively. Furthermore, we applied this procedure to analyze typical water samples, with enrichment factors of 85.29, 173.45 and 97.05 for βE2, E1 and DES, respectively, and good recoveries (≥ 89.82%). Altogether, the proposed procedure was simple and reliable, with potential application in large-scale water analysis.