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1.
Theor Appl Genet ; 137(9): 200, 2024 Aug 09.
Artículo en Inglés | MEDLINE | ID: mdl-39122841

RESUMEN

KEY MESSAGE: A stable Agrobacterium-mediated transformation system was constructed for B. juncea, and BjuLKP2 was overexpressed, leading to plant yellowing. A stable and efficient transformation system is necessary to verify gene functions in plants. To establish an Agrobacterium-mediated transformation system for B. juncea, various factors, including the explant types, hormone combination and concentration, infection time and concentration, were optimized. Eventually, a reliable system was established, and two BjuLKP2 overexpression (OE) lines, which displayed yellowing of cotyledons, shoot tips, leaves and flower buds, as well as a decrease in total chlorophyll content, were generated. qRT-PCR assays revealed significant upregulation of five chlorophyll synthesis genes and downregulation of one gene in the BjuLKP2 OE line. Furthermore, antioxidant capacity assays revealed reduced activities of APX, CAT and SOD, while POD activity increased in the BjuLKP2 OE26. Additionally, the kinetic determination of chlorophyll fluorescence induction suggested a decrease in the photosynthetic ability of BjuLKP2 OE26. GUS assays revealed the expression of BjuLKP2 in various tissues, including the roots, hypocotyls, cotyledons, leaf vasculature, trichomes, sepals, petals, filaments, styles and stigma bases, but not in seeds. Scanning electron revealed alterations in chloroplast ultrastructure in both the sponge and palisade tissue. Collectively, these findings indicate that BjuLKP2 plays a role in plant yellowing through a reduction in chlorophyll content and changes in chloroplasts structure.


Asunto(s)
Clorofila , Regulación de la Expresión Génica de las Plantas , Planta de la Mostaza , Agrobacterium/genética , Clorofila/metabolismo , Planta de la Mostaza/genética , Fotosíntesis , Hojas de la Planta/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Modificadas Genéticamente , Transformación Genética
2.
Medicine (Baltimore) ; 102(43): e35438, 2023 Oct 27.
Artículo en Inglés | MEDLINE | ID: mdl-37904411

RESUMEN

Using network pharmacology, molecular docking, and microRNA recognition, we have elucidated the mechanisms underlying the treatment of asthma by Jinxin oral liquid (JXOL). We began by identifying and normalizing the active compounds in JXOL through searches in the traditional Chinese medicine systems pharmacology database, SwissADME database, encyclopedia of traditional Chinese medicine database, HERB database, and PubChem. Subsequently, we gathered and standardized the targets of these active compounds from sources including the encyclopedia of traditional Chinese medicine database, similarity ensemble approach dataset, UniProt, and other databases. Disease targets were extracted from GeneCards, PharmGKB, OMIM, comparative toxicogenomics database, and DisGeNET. The intersection of targets between JXOL and asthma was determined using a Venn diagram. We visualized a Formula-Herb-Compound-Target-Disease network and a protein-protein interaction network using Cytoscape 3.9.0. Molecular docking studies were performed using Schrodinger software. To identify pathways related to asthma, we conducted gene ontology functional analysis and Kyoto encyclopedia of genes and genomes pathway enrichment analysis using Metascape. MicroRNAs regulating the hub genes were obtained from the miRTarBase database, and a network linking these targets and miRNAs was constructed. Finally, we found 88 bioactive components in JXOL and 218 common targets with asthma. Molecular docking showed JXOL key compounds strongly bind to HUB targets. According to gene ontology biological process analysis and Kyoto encyclopedia of genes and genomes pathway enrichment analysis, the PI3K-Akt signaling pathway, the MAPK signaling pathway, or the cAMP signaling pathway play a key role in treating of asthma by JXOL. The HUB target-miRNA network showed that 6 miRNAs were recognized. In our study, we have revealed for the first time the unique components, multiple targets, and diverse pathways in JXOL that underlie its mechanism of action in treating asthma through miRNAs.


Asunto(s)
Asma , Medicamentos Herbarios Chinos , MicroARNs , Humanos , Simulación del Acoplamiento Molecular , Farmacología en Red , Fosfatidilinositol 3-Quinasas , Asma/tratamiento farmacológico , Medicina Tradicional China , Medicamentos Herbarios Chinos/farmacología , Medicamentos Herbarios Chinos/uso terapéutico
3.
ACS Synth Biol ; 11(5): 1811-1823, 2022 05 20.
Artículo en Inglés | MEDLINE | ID: mdl-35481381

RESUMEN

Objectives: Emerging severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants pose a great threat and burden to global public health. Here, we evaluated a clustered regularly interspaced short palindromic repeat-associated enzyme 12a (CRISPR-Cas12a)-based method for detecting major SARS-CoV-2 variants of concern (VOCs) in SARS-CoV-2 positive clinical samples. Methods: Allele-specific CRISPR RNAs (crRNAs) targeting the signature mutations in the spike protein of SARS-CoV-2 are designed. A total of 59 SARS-CoV-2 positive oropharyngeal swab specimens were used to evaluate the performance of the CRISPR-Cas12a-mediated assay to identify major SARS-CoV-2 VOCs. Results: Compared with Sanger sequencing, the eight allele-specific crRNAs analyzed can specifically identify the corresponding mutations with a positive predictive value of 83.3-100% and a negative predictive value of 85.7-100%. Our CRISPR-Cas12a-mediated assay distinguished wild-type and four major VOCs (Alpha, Beta, Delta, and Omicron) of SARS-CoV-2 with a sensitivity of 93.8-100.0% and a specificity of 100.0%. The two methods showed a concordance of 98.3% (58/59) with a κ value of 0.956-1.000, while seven (11.9%) samples were found to be positive for extra mutations by the CRISPR-based assay. Furthermore, neither virus titers nor the sequences adjacent to the signature mutations were associated with the variation of fluorescence intensity detected or the false-positive reaction observed when testing clinical samples. In addition, there was no cross-reaction observed when detecting 33 SARS-CoV-2 negative clinical samples infected with common respiratory pathogens. Conclusions: The CRISPR-Cas12a-based genotyping assay is highly sensitive and specific when detecting both the SARS-CoV-2 wild-type strain and major VOCs. It is a simple and rapid assay that can monitor and track the circulating SARS-CoV-2 variants and the dynamics of the coronavirus disease 2019 (COVID-19) pandemic and can be easily implemented in resource-limited settings.


Asunto(s)
COVID-19 , SARS-CoV-2 , COVID-19/diagnóstico , Sistemas CRISPR-Cas/genética , Humanos , Mutación , SARS-CoV-2/genética
4.
Plant Physiol Biochem ; 179: 25-31, 2022 May 15.
Artículo en Inglés | MEDLINE | ID: mdl-35306327

RESUMEN

Brassica juncea var. tumida Tsen et Lee (Tumorous stem mustard) is an unique vegetable in China. Its enlarged tumorous stem was used as main raw material to produce pickle (Zhacai). In practice, early-bolting happens around 15% of planting area all year and inhibits its production. Here, about 209 PP2C proteins were identified through HMMER software and divided into 13 sub-families in B. juncea. BjuPP2C52 belongs to E sub-family, was up-regulated at reproductive growth stages and interacts with BjuFKF1, a key protein in regulating plant photoperiod flowering, in vitro and in vivo. To explore interactive proteins, BjuPP2C52 was used as bait, 12 potential interactive proteins were screened from yeast library, and they are BjuCOL3, BjuCOL5, BjuAP2, BjuAP2-1, BjuSVP-1, BjuFLC-2, BjuSKP1f, BjuA014572, BjuA008686, BjuO002119, BjuB036787 and BjuA019268. Further study verified that 10 out of the 12 screened proteins interacted with BjuPP2C52 in vivo. qRT-PCR was conducted to understand the expression pattern of those 10 interactive proteins in different tissues and development stages in B. juncea. The results showed that BjuCOL3, BjuCOL5, BjuB036787 and BjuA019268 were significantly up-regulated, while BjuA008686 and BjuO002119 were down-regulated in flowers compared with other four tissues. In developmental stages, BjuCOL5, BjuAP2, BjuAP2-1, BjuA014572, BjuB036787 and BjuA019268 were significantly up-regulated, while BjuSVP-1, BjuA008686 and BjuO002119 were down-regulated at reproductive stages. Based on the results, BjuCOL5, BjuAP2, BjuAP2-1, BjuSVP-1, BjuA014572, BjuB036787 and BjuA019268 may function in regulating flowering time in B. juncea.


Asunto(s)
Regulación de la Expresión Génica de las Plantas , Planta de la Mostaza , Flores/metabolismo , Humanos , Filogenia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo
5.
3 Biotech ; 10(5): 222, 2020 May.
Artículo en Inglés | MEDLINE | ID: mdl-32368431

RESUMEN

Genic male sterility (GMS) is a common and important trait, which is widely used for the production of hybrid seeds. However, the molecular mechanism of GMS in watermelon remains poorly understood. In this study, we comparatively analyzed the transcriptome profiles of sterile and fertile floral buds using the bulked segregant analysis (BSA) and transcriptome sequencing (RNA-seq). A total of 2507 differentially expressed genes (DEGs) including 593 up-regulated and 1914 down-regulated, were identified to be related to male sterility in watermelon line Se18. Gene ontology (GO) analysis showed that 57 GO terms were significantly enriched, while Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis revealed plant hormone signal transduction, glycolysis/gluconeogenesis, starch and sucrose metabolism, plant-pathogen interaction, phenylpropanoid biosynthesis pathways were obviously enriched. Furthermore, the efficiency of the RNA-seq analysis was validated by quantitative real-time PCR (qRT-PCR). Among the DEGs, some valuable candidate genes involved in pollen development were identified, such as gene Cla000029, a bHLH transcription factor and homologous to MS1 in Arabidopsis. Moreover, other DEGs including MYB gene Cla012590 (MYB26), Cla017100 (MYB21), etc., also provide useful information for further understanding the function of key genes involved in pollen development. This study provides new insights into the global network of male sterility in watermelon.

6.
Plant Sci ; 278: 32-43, 2019 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-30471727

RESUMEN

Drought stress has become one of the most urgent environmental hazards for horticultural crops. In this research, we analyzed watermelon adaptation strategies to drought stress in drought-tolerant (M20) and -susceptible (Y34) genotypes via transcriptomic and physiological analyses. After drought stress, a total of 6228 and 4311 differentially expressed genes (DEGs) were observed in Y34 and M20, respectively. Numerous DEGs were involved in various defense responses such as antioxidation, protein protection, osmotic adjustment, wax accumulation, hormone signaling, and melatonin biosynthesis. Accordingly, the contents of ABA, melatonin, wax, and some osmoprotectants were increased by drought stress in both Y34 and especially M20. Exogenous application of melatonin or ABA induced wax accumulation and drought tolerance and melatonin may function upstream of ABA. In comparison to Y34, M20 was more able to activate ABA signaling, melatonin biosynthesis, osmotic adjustment, antioxidation, and wax accumulation under drought stress. These stronger responses confer M20 tolerance to drought. Photosynthesis and most DEGs involved in photosynthesis and cell growth were decreased by drought stress in both M20 and especially Y34. For drought-susceptible genotypes, growth retardation may be an important mechanism for saving and redistributing resources in order to reprogram stress signaling networks.


Asunto(s)
Citrullus/metabolismo , Estrés Fisiológico , Agua/metabolismo , Adaptación Biológica/genética , Ácido Ascórbico/metabolismo , Respiración de la Célula , Clorofila/metabolismo , Citrullus/genética , Sequías , Perfilación de la Expresión Génica , Genotipo , Glutatión/metabolismo , Melatonina/metabolismo , Fotosíntesis , Reguladores del Crecimiento de las Plantas/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Prolina/metabolismo
7.
PLoS One ; 13(1): e0191308, 2018.
Artículo en Inglés | MEDLINE | ID: mdl-29338040

RESUMEN

Despite identification of WRKY family genes in numerous plant species, a little is known about WRKY genes in watermelon, one of the most economically important fruit crops around the world. Here, we identified a total of 63 putative WRKY genes in watermelon and classified them into three major groups (I-III) and five subgroups (IIa-IIe) in group II. The structure analysis indicated that ClWRKYs with different WRKY domains or motifs may play different roles by regulating respective target genes. The expressions of ClWRKYs in different tissues indicate that they are involved in various tissue growth and development. Furthermore, the diverse responses of ClWRKYs to drought, salt, or cold stress suggest that they positively or negatively affect plant tolerance to various abiotic stresses. In addition, the altered expression patterns of ClWRKYs in response to phytohormones such as, ABA, SA, MeJA, and ETH, imply the occurrence of complex cross-talks between ClWRKYs and plant hormone signals in regulating plant physiological and biological processes. Taken together, our findings provide valuable clues to further explore the function and regulatory mechanisms of ClWRKY genes in watermelon growth, development, and adaption to environmental stresses.


Asunto(s)
Citrullus/genética , Citrullus/fisiología , Regulación de la Expresión Génica de las Plantas , Proteínas de Plantas/genética , Estrés Fisiológico/genética , Factores de Transcripción/genética , Secuencias de Aminoácidos , Secuencia de Aminoácidos , Citrullus/efectos de los fármacos , Secuencia Conservada , Evolución Molecular , Duplicación de Gen , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Filogenia , Reguladores del Crecimiento de las Plantas/farmacología , Proteínas de Plantas/química , Alineación de Secuencia , Estrés Fisiológico/efectos de los fármacos , Sintenía , Factores de Transcripción/química
8.
PLoS One ; 12(4): e0176352, 2017.
Artículo en Inglés | MEDLINE | ID: mdl-28437432

RESUMEN

The calcium-dependent protein kinase (CDPK) is a ser/thr protein kinase that plays vital roles in plant growth, development, and responses to multiple stresses. Despite an important member of the stress responsive gene family, little is known about the evolutionary history and expression patterns of CDPK genes in melon. Herein, a total of 18 CDPK genes and 7 CDPK-related protein kinases (CRK) genes were identified in the melon genome via bioinformatic analysis, which were unevenly distributed across eleven chromosomes with an apparent exception for chromosome 3. Comparative syntenic analysis between Cucumis melo L. and Arabidopsis thaliana revealed that 13 CmCDPKs and 19 AtCPKs existed in 20 corresponding syntenic blocks. In addition, based on gene structure and phylogenetic analyses, all CmCDPKs were divided into four groups (CDPK I-IV) and CmCRKs clustered into one group (CRK I). Interestingly, group CDPK IV was clearly distinct from the other three CDPK groups, but clustered with CRK I on the phylogenetic tree, implying their origination from a common ancestor. Furthermore, CmCDPKand CmCRK genes were differentially expressed in response to various stimuli, such as biotic stress (Podosphaera xanthii), abiotic stress (salt and cold), and hormone (abscisic acid) treatment. To our knowledge, this is the first report on CDPK and CRK gene families in melon, which provides a basic foundation for functional characterizations of CmCDPK and CmCRK genes in the future.


Asunto(s)
Cucumis melo/genética , Regulación de la Expresión Génica de las Plantas , Proteínas de Plantas/genética , Proteínas Quinasas/genética , Expresión Génica , Perfilación de la Expresión Génica , Genoma de Planta , Filogenia , Estrés Fisiológico/genética
9.
PLoS One ; 11(11): e0166314, 2016.
Artículo en Inglés | MEDLINE | ID: mdl-27861528

RESUMEN

Osmotic stress adversely affects the growth, fruit quality and yield of watermelon (Citrullus lanatus (Thunb.) Matsum. & Nakai). Increasing the tolerance of watermelon to osmotic stress caused by factors such as high salt and water deficit is an effective way to improve crop survival in osmotic stress environments. Roots are important organs in water absorption and are involved in the initial response to osmosis stress; however, few studies have examined the underlying mechanism of tolerance to osmotic stress in watermelon roots. For better understanding of this mechanism, the inbred watermelon accession M08, which exhibits relatively high tolerance to water deficits, was treated with 20% polyethylene glycol (PEG) 6000. The root samples were harvested at 6 h after PEG treatment and untreated samples were used as controls. Transcriptome analyses were carried out by Illumina RNA sequencing. A total of 5246 differentially expressed genes were identified. Gene ontology enrichment and biochemical pathway analyses of these 5246 genes showed that short-term osmotic stress affected osmotic adjustment, signal transduction, hormone responses, cell division, cell cycle and ribosome, and M08 may repress root growth to adapt osmotic stress. The results of this study describe the watermelon root transcriptome under osmotic stress and propose new insight into watermelon root responses to osmotic stress at the transcriptome level. Accordingly, these results allow us to better understand the molecular mechanisms of watermelon in response to drought stress and will facilitate watermelon breeding projects to improve drought tolerance.


Asunto(s)
Citrullus/genética , Perfilación de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Presión Osmótica , Raíces de Plantas/genética , Estrés Fisiológico/genética , Transcriptoma , Citrullus/metabolismo , Biología Computacional , Ontología de Genes , Secuenciación de Nucleótidos de Alto Rendimiento , Raíces de Plantas/metabolismo , Especies Reactivas de Oxígeno , Factores de Tiempo
10.
PLoS One ; 11(6): e0156515, 2016.
Artículo en Inglés | MEDLINE | ID: mdl-27258145

RESUMEN

Continuous cropping of watermelon (Citrullus lanatus L.) can lead to reduced yield and quality. We aimed to determine the effects of cattle manure addition and rotation with green garlic to improve yield and reduce disease incidence in watermelon and to examine the effects on the biological and chemical characteristics of the soil. Field experiments were performed during 2012-2014 on land previously under two years of continuous watermelon cropping in northwest China. We examined three treatment combinations: watermelon and garlic rotation, cattle manure application before watermelon planting, and combined cattle manure addition and crop rotation. Watermelon monoculture was retained as a control. Watermelon yield was significantly higher and disease incidence was lower in the treatments than the control. The populations of soil bacteria and actinomycetes and the bacteria/fungi ratio increased significantly and soil enzyme activities were generally enhanced under treatments. Available nutrients and soil organic matter contents were much higher under experimental treatments than the control. Results suggest both cattle manure application and garlic rotation can ameliorate the negative effects of continuous cropping. The combined treatment of cattle manure addition and green garlic rotation was optimal to increase yield, reduce disease incidence and enhance soil quality.


Asunto(s)
Citrullus/crecimiento & desarrollo , Ajo/crecimiento & desarrollo , Microbiología del Suelo , Agricultura/métodos , Animales , Bacterias/aislamiento & purificación , Bovinos , China , Hongos/aislamiento & purificación
11.
Front Plant Sci ; 7: 644, 2016.
Artículo en Inglés | MEDLINE | ID: mdl-27242845

RESUMEN

Drought stress has become an increasingly serious environmental issue that influences the growth and production of watermelon. Previous studies found that arbuscular mycorrhizal (AM) colonization improved the fruit yield and water use efficiency (WUE) of watermelon grown under water stress; however, the exact mechanisms remain unknown. In this study, the effects of Glomus versiforme symbiosis on the growth, physio-biochemical attributes, and stress-responsive gene expressions of watermelon seedlings grown under well-watered and drought conditions were investigated. The results showed that AM colonization did not significantly influence the shoot growth of watermelon seedlings under well-watered conditions but did promote root development irrespective of water treatment. Drought stress decreased the leaf relative water content and chlorophyll concentration, but to a lesser extent in the AM plants. Compared with the non-mycorrhizal seedlings, mycorrhizal plants had higher non-photochemical quenching values, which reduced the chloroplast ultrastructural damage in the mesophyll cells and thus maintained higher photosynthetic efficiency. Moreover, AM inoculation led to significant enhancements in the enzyme activities and gene expressions of superoxide dismutase, catalase, ascorbate peroxidase, glutathione reductase, and monodehydroascorbate reductase in watermelon leaves upon drought imposition. Consequently, AM plants exhibited lower accumulation of MDA, H2O2 and [Formula: see text] compared with non-mycorrhizal plants. Under drought stress, the soluble sugar and proline contents were significantly increased, and further enhancements were observed by pre-treating the drought-stressed plants with AM. Taken together, our findings indicate that mycorrhizal colonization enhances watermelon drought tolerance through a stronger root system, greater protection of photosynthetic apparatus, a more efficient antioxidant system and improved osmoregulation. This study contributes to advances in the knowledge of AM-induced drought tolerance.

12.
Zhonghua Yu Fang Yi Xue Za Zhi ; 49(7): 615-20, 2015 Jul.
Artículo en Chino | MEDLINE | ID: mdl-26310474

RESUMEN

OBJECTIVE: To analyze epidemiological characteristics of norovirus variant of GII.4 Sydney from January 2012 to June 2014 in sentinel hospitals of Guangdong province, as well as the outbreaks caused by norovirus variant of GII.4 Sydney. METHODS: During January 2012 to June 2014, a total of 10 750 fecal samples were obtained from 22 hospitals of surveillance sites in Guangdong province. Those samples were sent to the local municipal CDCs for extracting and detecting norovirus nucleic acid. Then, all the positive samples were delivered to Guangdong provincial CDC that used Random Number Method to draw 855 positive samples for norovirus genotyping, and 690 samples were successfully sequenced. Chi-square tests were used to compare norovirus infection status of diarrhea cases in different age groups as well as during different periods. Epidemiological data of 13 outbreaks which were caused by norovirus variant of GII.4 Sydney from January 2012 to June 2014 were collected from the Public Health Emergency Management Information System of Guangdong Province, and the epidemiological characteristics were analyzed. RESULTS: The norovirus variant of GII.4 Sydney was first detected in August 2012 and the detection rate was 13/15 in November 2012. During November 2012 to January 2013 (period T1), the norovirus positive rate of each month was 23.8% (100/421), 15.9% (61/383) and 19.2% (95/495), respectively. During November 2013 to January 2014 (period T2), the norovirus positive rate of each month was 17.0% (90/529), 8.7% (37/426) and 11.2% (46/409), respectively which were significantly lower than that of period T1 (χ² alue was 6.65, 9.93 and 10.74. P value was 0.010, 0.002, and 0.001, respectively). In period T1, the norovirus positive rate of people ages 15 and older was 26.3% (143/543) and the rate of people under 15 was 14.9% (113/756) (χ² = 2.90, P < 0.001). In period T2, the norovirus positive rate of people ages 15 and older was 10.1% (52/516) and the rate of people under 15 (14.3% (121/848)) (χ²= 5.09, P = 0.024). The foodborne transmission was the infection source for ten of thirteen outbreaks. CONCLUSION: The norovirus variant of GII.4 Sydney was first detected in August 2012. The epidemic began to occur in the community since November 2012, and the strength of the epidemic declined 1 year later. The foodborne transmission was the main infection sources for the outbreaks caused by norovirus variant of GII.4 Sydney.


Asunto(s)
Epidemias , Enfermedades Transmitidas por los Alimentos , Genotipo , Norovirus , Vigilancia de Guardia , Adolescente , Adulto , Ácido Quenodesoxicólico/análogos & derivados , Niño , China , Diarrea , Brotes de Enfermedades , Humanos
13.
Emerg Infect Dis ; 21(7): 1240-2, 2015 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-26080037

RESUMEN

In the past decade, the most prevalent norovirus genotype causing viral gastroenteritis outbreaks worldwide, including China, has been GII.4. In winter 2014-15, norovirus outbreaks in Guangdong, China, increased. Sequence analysis indicated that 82% of the outbreaks were caused by a norovirus GII.17 variant.


Asunto(s)
Brotes de Enfermedades , Gastroenteritis/epidemiología , Gastroenteritis/virología , Norovirus/genética , China/epidemiología , Genes Virales , Genotipo , Humanos , Datos de Secuencia Molecular , Análisis de Secuencia de ADN
14.
Appl Environ Microbiol ; 81(7): 2311-9, 2015 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-25616804

RESUMEN

An aseptic meningitis outbreak occurred in Luoding City of Guangdong, China, in 2012, and echovirus type 30 (ECHO30) was identified as the major causative pathogen. Environmental surveillance indicated that ECHO30 was detected in the sewage of a neighboring city, Guangzhou, from 2010 to 2012 and also in Luoding City sewage samples (6/43, 14%) collected after the outbreak. In order to track the potential origin of the outbreak viral strains, we sequenced the VP1 genes of 29 viral strains from clinical patients and environmental samples. Sequence alignments and phylogenetic analyses based on VP1 gene sequences revealed that virus strains isolated from the sewage of Guangzhou and Luoding cities matched well the clinical strains from the outbreak, with high nucleotide sequence similarity (98.5% to 100%) and similar cluster distribution. Five ECHO30 clinical strains were clustered with the Guangdong environmental strains but diverged from strains from other regions, suggesting that this subcluster of viruses most likely originated from the circulating virus in Guangdong rather than having been more recently imported from other regions. These findings underscore the importance of long-term, continuous environmental surveillance and genetic analysis to monitor circulating enteroviruses.


Asunto(s)
Brotes de Enfermedades , Infecciones por Echovirus/epidemiología , Infecciones por Echovirus/virología , Enterovirus Humano B/clasificación , Monitoreo del Ambiente , Meningitis Aséptica/epidemiología , Meningitis Aséptica/virología , China/epidemiología , Ciudades , Análisis por Conglomerados , Infecciones por Echovirus/transmisión , Enterovirus Humano B/genética , Enterovirus Humano B/aislamiento & purificación , Humanos , Epidemiología Molecular , Datos de Secuencia Molecular , Filogenia , ARN Viral/genética , Alineación de Secuencia , Análisis de Secuencia de ADN , Aguas del Alcantarillado/virología , Proteínas Estructurales Virales/genética
15.
PLoS One ; 9(1): e86877, 2014.
Artículo en Inglés | MEDLINE | ID: mdl-24497989

RESUMEN

Enterovirus 96 (EV-C96) is a newly described serotype within the enterovirus C (EV-C) species, and its biological and pathological characters are largely unknown. In this study, we sequenced the whole genome of a novel EV-C96 strain that was isolated in 2011 from a patient with acute flaccid paralysis (AFP) in Guangdong province, China and characterized the properties of its infection. Sequence analysis revealed the close relationship between the EV-C96 strains isolated from the Guangdong and Shandong provinces of China, and suggested that recombination events occurred both between these EV-C96 strains and with other EV-C viruses. Moreover, the virus replication kinetics showed EV-C96 Guangdong strain replicated at a high rate in RD cells and presented a different cell tropism to other strains isolated from Shandong recently. These findings gave further insight into the evolutionary processes and extensive biodiversity of EV-C96.


Asunto(s)
Enterovirus Humano C/genética , Infecciones por Enterovirus/virología , Paraplejía/virología , Animales , Línea Celular Tumoral , Preescolar , China , Chlorocebus aethiops , Enterovirus Humano C/aislamiento & purificación , Enterovirus Humano C/fisiología , Femenino , Genoma Viral , Humanos , Cinética , Filogenia , Recombinación Genética , Análisis de Secuencia de ADN , Células Vero , Proteínas Estructurales Virales/genética , Replicación Viral
16.
Appl Environ Microbiol ; 79(24): 7679-83, 2013 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-24096418

RESUMEN

The human-pathogenic viruses in urban sewage have been extensively monitored to obtain information on circulating viruses in human communities. Enteroviruses (EVs) excreted by patients who present with diverse clinical syndromes can remain infectious in the environment for several weeks, and limited data on circulating environmental EVs are available. A 4-year (2009 to 2012) surveillance study was conducted to detect nonpolio enteroviruses (NPEVs) in the urban sewage of Guangzhou city, China. After the viruses in the sewage samples were concentrated and isolated, molecular identification was used to detect and type the NPEVs. During the 4-year study, 17 different NPEV serotypes were identified in the sewage of Guangzhou city. The most common serotypes were echovirus 11 (ECHO11), ECHO6, ECHO7, and ECHO12 and coxsackie group B viruses 5 (CVB5) and CVB3. The predominant serotypes were influenced by spatial and temporal factors and differed each year. CVB5 was commonly detected in 2009 and 2010 but was rarely isolated in 2011 and 2012. In contrast, CVB3 was not observed in 2009 and 2010 but was increasingly detected in 2011 and 2012. Our study provides an overview of the serotype distribution and circulation patterns of NPEVs in the sewage of Guangzhou, China. In the absence of a systematic EV disease surveillance system, the detection and characterization of sewage-borne NPEVs will help us better understand the changes in EV disease trends and the epidemic background of circulating EVs, which could help interpret the EV trends and warn of future outbreaks in this area.


Asunto(s)
Enterovirus/clasificación , Enterovirus/aislamiento & purificación , Aguas del Alcantarillado/virología , China , Enterovirus/genética , Genotipo , Humanos , Prevalencia , ARN Viral/genética , Serotipificación
17.
Nan Fang Yi Ke Da Xue Xue Bao ; 33(4): 568-72, 2013 Apr.
Artículo en Chino | MEDLINE | ID: mdl-23644122

RESUMEN

OBJECTIVE: To explore a convenient and effective method for norovirus nucleic acid extraction from oysters suitable for long-term viral surveillance. METHODS: Two methods, namely method A (glycine washing and polyethylene glycol precipitation of the virus followed by silica gel centrifugal column) and method B (protease K digestion followed by application of paramagnetic silicon) were compared for their performance in norovirus nucleic acid extraction from oysters. Real-time RT-PCR was used to detect norovirus in naturally infected oysters and in oysters with induced infection. RESULTS: The two methods yielded comparable positive detection rates for the samples, but the recovery rate of the virus was higher with method B than with method A. CONCLUSION: Method B is a more convenient and rapid method for norovirus nucleic acid extraction from oysters and suitable for long-term surveillance of norovirus.


Asunto(s)
Centrifugación/métodos , Norovirus/aislamiento & purificación , Ostreidae/virología , ARN Viral/aislamiento & purificación , Animales , Norovirus/genética
18.
Zhonghua Liu Xing Bing Xue Za Zhi ; 34(10): 989-92, 2013 Oct.
Artículo en Chino | MEDLINE | ID: mdl-24377993

RESUMEN

OBJECTIVE: To analyze the epidemiological and etiological characteristics of diarrheal disease among children under 5 years of age in Guangdong province, in 2012. METHODS: 64 hospitals in 21 cities were chosen as the diarrheal syndromic surveillance sites, of which 14 hospitals were selected to carry out etiological surveillance among children under 5 years of age, including isolation and culture of both Vibrio cholera and Shigella as well as nucleic acid detection of rotavirus and norovirus by PCR. Descriptive method was used to analyze data from syndromic and etiological surveillance programs on diarrheal, from 1932 parents of the children. RESULTS: In 2012, the outpatient attendance rate on diarrheal among children under 5 years was 0.8%. The proportion of diarrheal in children under 5-year-olds was 63.5%, among the total number of diarrheal outpatients at the outpatient clinics under surveillance program. The morbidity of infectious diarrhea was 1454.5/10 million in children under 5 years of age. A total number of 1932 specimens were collected from children under 5 years of age, in the outpatient department. Among these specimens,Vibrio cholera appeared all negative but one was Shigella positive and proved to be Sh. sonnei. The positive rates of rotavirus and norovirus were 14.1% (273/1932)and 16.9% (326/1932). Both rotavirus and norovirus were found in 24 specimens, with a positive rate as 1.2% . 112 specimens were successfully gene sequenced for rotavirus, of which 33.9% as G1[P8] genotype, 25.9% as G9[P8], 12.5% as G2[P4] and 9.8% as G3[P8] respectively. 90 specimens were successfully gene-sequenced for norovirus, of which 76.7% as G II.4 genotype. Genetic subtypes of G II. 4/2006b, accounted for 50.0% and could be detected around the year except for June and December. New G II. 4/Sydney Strain_2012 was first detected in August and became the predominant in December. In addition, 5 specimens belonged to G I genotype with other 16 subtypes of G II. CONCLUSION: Results from our study proved that children under 5 years of age belonged to high-risk group for diarrheal disease in Guangdong province. Rotavirus and norovirus were both diverse in terms of genome.


Asunto(s)
Diarrea/epidemiología , Diarrea/microbiología , Infecciones por Caliciviridae/epidemiología , Preescolar , China/epidemiología , Diarrea/virología , Femenino , Genotipo , Humanos , Lactante , Masculino , Infecciones por Rotavirus/epidemiología
19.
APMIS ; 120(8): 683-8, 2012 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-22779692

RESUMEN

Acute respiratory infections (ARI) are the major worldwide health problem due to associated high morbidity and mortality rates. Adenovirus (Adv) is one of the most common causes of viral ARI, and thus calls for specific diagnosis and better understanding of the epidemiology and clinical characteristics. Our aims were to find out the status of Adv infection in children <14 years with ARI, analyze the epidemiology and clinical characteristics among the Adv-infected children in Guangzhou, China, and to provide some basis for the research of Adv. The throat and pharyngeal swabs were collected among the children with acute respiratory tract infections in outpatient department from September 2006 to August 2008. The samples were analyzed by PCR and the sequences were blasted with the sequences of Adv in GenBank. Clinical data were analyzed along with virological data by using appropriate statistical methods. Adv was detected in 25 out of 512 (4.9%) children. The genome types of 23 samples were determined after analysis of the gene sequence. The most prevalent Adv type was species B type 3. Among the patients, 10 were of Ad3 (43.5%), three were of Ad1 (1.3%), five were of species C Ad2 (21.7%), and five were of species E Ad4 (21.7%). A higher incidence of positive results was found during the summer season, thus showing a pattern of seasonality. There exists Adv infection in children with acute respiratory system diseases in Guangzhou area. No significant differences were found among different age groups and gender groups. Co-infections with other respiratory virus were detected in 64% of the Adv positive samples.


Asunto(s)
Adenoviridae/aislamiento & purificación , Infecciones por Adenovirus Humanos/epidemiología , Infecciones por Adenovirus Humanos/virología , Infecciones del Sistema Respiratorio/epidemiología , Infecciones del Sistema Respiratorio/virología , Adenoviridae/genética , Adolescente , Niño , Preescolar , China , Femenino , Genoma Viral , Humanos , Lactante , Recién Nacido , Masculino , Filogenia
20.
Zhonghua Yu Fang Yi Xue Za Zhi ; 45(9): 825-9, 2011 Sep.
Artículo en Chino | MEDLINE | ID: mdl-22177307

RESUMEN

OBJECTIVE: To investigate the pathogens of acute respiratory infection (ARI) in Guangzhou from 2006 to 2009. METHODS: A total of 1554 cases of ARI patients in Second Affiliated Hospital of Guangzhou Medical College from September 2006 to September 2009, were recruited in the survey. The sample of throat and pharyngeal swab were collected from each patient.11 types of virus including influenza A (FluA), influenza B (FluB), adenovirus (ADV), human rhinovirus (HRV), respiratory syncytial virus (RSV), parainfluenza virus type 1, type 2, type 3 (HPIV1, HPIV2, HPIV3), human metapneumovirus (MPV) and human coronavirus (HCoV) type 229E, type OC43 were detected by Fluorescence Quota PCR method. The epidemic feature and clinical characteristic of each virus were then analyzed. RESULTS: Virus were found in 1024 samples in total, accounting for 65.9% (1024/1554). RSV was the most common virus, which was found in 261 samples (16.8%); and followed by HRV as 13.9% (216/1554), FluA as 11.6% (181/1554), MPV as 6.5% (101/1554), FluB as 6.4% (99/1554), HPIV as 4.9% (76/1554), ADV as 3.5% (55/1554) and HCoV as 2.3% (35/1554). HPIV and HCoV shared a similar infection ratio among different age groups. The infection ratio of FluA and FluB was highest among 15-24 years old group, accounting for 16.5% (29/176) and 7.4% (13/176) respectively. MPV, RSV and HRV were the main pathogens caused infection among children under 4 years old, accounting for 9.7% (49/503), 21.7% (109/503) and 18.9% (95/503). The infection ratio of ADV was 6.0% (19/318), which was the most common pathogen among 5-14 years old patients. The incidence rate of HPIV and HRV showed no obvious seasonal features; while the prevalence of FluA, FluB, RSV, ADV, MPV and HCoV changed significantly in different seasons.22.2% (227/1024) ARI patients co-infected other respiratory virus.90.1% (163/181) FluA patients, 88.9% (88/99) FluB patients and 92.7% (51/55) ADV patients had high fever symptoms. CONCLUSION: RSV was the main pathogen of ARI, and the new-found virus MPV was also another crucial pathogen. Some pathogens' incidence rate were related to the season and patient's age. Co-infections of other respiratory virus were also detected in parts of ARI patients.


Asunto(s)
Metapneumovirus/aislamiento & purificación , Infecciones del Sistema Respiratorio/epidemiología , Infecciones del Sistema Respiratorio/virología , Enfermedad Aguda , Adolescente , Adulto , Anciano , Niño , Preescolar , China/epidemiología , Humanos , Lactante , Recién Nacido , Persona de Mediana Edad , Virus Sincitiales Respiratorios/aislamiento & purificación , Adulto Joven
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