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Human respiratory syncytial virus (HRSV) is the most prevalent pathogen contributing to acute respiratory tract infections (ARTI) in infants and young children and can lead to significant financial and medical costs. Here, we developed a simultaneous, dual-gene and ultrasensitive detection system for typing HRSV within 60 minutes that needs only minimum laboratory support. Briefly, multiplex integrating reverse transcription-recombinase polymerase amplification (RT-RPA) was performed with viral RNA extracted from nasopharyngeal swabs as a template for the amplification of the specific regions of subtypes A (HRSVA) and B (HRSVB) of HRSV. Next, the Pyrococcus furiosus Argonaute (PfAgo) protein utilizes small 5'-phosphorylated DNA guides to cleave target sequences and produce fluorophore signals (FAM and ROX). Compared with the traditional gold standard (RT-qPCR) and direct immunofluorescence assay (DFA), this method has the additional advantages of easy operation, efficiency and sensitivity, with a limit of detection (LOD) of 1 copy/µL. In terms of clinical sample validation, the diagnostic accuracy of the method for determining the HRSVA and HRSVB infection was greater than 95%. This technique provides a reliable point-of-care (POC) testing for the diagnosis of HRSV-induced ARTI in children and for outbreak management, especially in resource-limited settings.
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RNA Viral , Infecções por Vírus Respiratório Sincicial , Vírus Sincicial Respiratório Humano , Sensibilidade e Especificidade , Humanos , Vírus Sincicial Respiratório Humano/genética , Vírus Sincicial Respiratório Humano/isolamento & purificação , Infecções por Vírus Respiratório Sincicial/diagnóstico , Infecções por Vírus Respiratório Sincicial/virologia , RNA Viral/genética , Lactente , Pyrococcus furiosus/genética , Pyrococcus furiosus/isolamento & purificação , Proteínas Argonautas/genética , Técnicas de Diagnóstico Molecular/métodos , Técnicas de Amplificação de Ácido Nucleico/métodos , Limite de Detecção , Nasofaringe/virologia , Infecções Respiratórias/diagnóstico , Infecções Respiratórias/virologia , Pré-EscolarRESUMO
This study evaluated the anti-diabetic effect of polysaccharides isolated from Ornithogalum caudatum and their underlying mechanisms. To achieve this, a type 2 diabetes mellitus mouse model was established using a combination of a high-fat diet and low-dose streptozotocin injection. The mice were treated with Ornithogalumcaudatum polysaccharides (OCPs) for 4 weeks. OCPs treatment significantly decreased body weight loss, fasting blood glucose levels, and plasma insulin levels in diabetic mice. Additionally, compared with the untreated group, OCPs treatment significantly decreased total cholesterol, triacylglycerol, and low-density lipoprotein-cholesterol levels, but increased those of high-density lipoprotein-cholesterol in diabetic mice. Moreover, antioxidant enzyme activity and histopathology results revealed that OCPs effectively alleviated oxidative stress and streptozotocin-induced lesions by increasing antioxidant enzyme activity. Results from mechanistic studies showed that OCPs treatment significantly increased the expression of p-PI3K, p-Akt, and p-GSK-3ß in the liver. Moreover, OCPs optimized the gut microbiota composition of diabetic mice by significantly decreasing the Firmicutes/Bacteroidetes ratio and increasing the levels of beneficial bacteria (Muribaculaceae_norank, Prevotellaceae_UCG-001 and Alloprevotella). Overall, these findings suggest that OCPs exert anti-diabetic effects by triggering the PI3K/Akt/GSK-3ß signaling pathway and regulating the gut microbiota.
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BACKGROUND: Since January 2020, measures has been adopted in the Chaoshan area to limit the spread of COVID-19. Restrictions were removed after August 2020. At the same time, children returned to school. We previously reported the changes of 14 main respiratory pathogens in hospitalized children before and during the COVID-19 outbreak in Chaoshan area. However, the changes of respiratory pathogen spectrum in hospitalized children after the epidemic are still unknown, which will be elucidated in this study. METHODS: There are 6201 children hospitalized with respiratory tract infection were enrolled in the study, which were divided into two groups: 2533 from outbreak group (1 January 2020-31 December 2020), and 3668 from post-outbreak group (1 January 2021-31 December 2021). Pharyngeal swab samples were collected. 14 respiratory tract pathogens were detected by liquid chip technology. RESULTS: The positive rate of pathogen detection is significantly lower in the outbreak group (65.42%, 1657/2533) than that in the post-outbreak group (70.39%, 2582/3668; χ2 = 17.15, P < 0.05). The Influenza A virus (FluA) detection rate was 1.9% (49) in 2020, but 0% (0) in 2021. The detection rates of Bordetella pertussis (BP) decreased from 1.4% (35) in 2020 to 0.5% (17) in 2021. In contrast, the detection rates of Influenza B virus (FluB), Cytomegalovirus (CMV), Haemophilus influenzae (HI), Streptococcus pneumoniae (SP) increased from 0.3% (8), 24.7% (626), 2.0% (50) and 19.4% (491) in 2020 to 3.3% (121), 27.9% (1025), 4.6% (169), 22.8% (836) in 2021, respectively (P < 0.01). CONCLUSIONS: The detection rates of pathogens such as FluA, FluB, CMV, HI, SP, BP were statistically different between 2020 and 2021. From 2020 to 2021, the positive rates of Flu, CMV, HI and SP increased, while the positive rates of FluA and BP decreased. After the COVID-19 prevention and control measures are gradually relaxed, the positive rate of respiratory pathogens in children aged from 6 months to 6 years will increase.
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COVID-19 , Infecções por Citomegalovirus , Infecções Respiratórias , Criança , Humanos , Lactente , Criança Hospitalizada , COVID-19/epidemiologia , Infecções Respiratórias/epidemiologia , Surtos de Doenças , Citomegalovirus , Infecções por Citomegalovirus/epidemiologiaRESUMO
BACKGROUND: The epidemiology of local viral etiologies is essential for the management of viral respiratory tract infections. Limited data are available in China to describe the epidemiology of viral respiratory infections, especially in small-medium cities and rural areas. OBJECTIVES: To determine the viral etiology and seasonality of acute respiratory infections in hospitalized children, a 3-year study was conducted in Shenzhen, China. METHODS: Nasopharyngeal aspirates from eligible children were collected. Influenza and other respiratory viruses were tested by molecular assays simultaneously. Data were analyzed to describe the frequency and seasonality. RESULTS: Of the 2025 children enrolled in the study, 971 (48.0%) were positive for at least one viral pathogen, in which 890 (91.7%) were <4 years of age. The three most prevalent viruses were influenza A (IAV; 35.8%), respiratory syncytial virus (RSV; 30.5%) and human rhinovirus (HRV; 21.5%). Co-infections were found in 302 cases (31.1%), and dual viral infection was dominant. RSV, HRV and IAV were the most frequent viral agents involved in co-infection. On the whole, the obvious seasonal peaks mainly from March to May were observed with peak strength varying from 1 year to another. CONCLUSIONS: This study provides a basic profile of the epidemiology of acute respiratory viral infection in hospitalized children in Shenzhen. The spectrum of viruses in the study site is similar to that in other places, but the seasonality is closely related to geographic position, different from that in big cities in northern China and neighboring Hong Kong.
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Hospitalização , Infecções Respiratórias/epidemiologia , Viroses/epidemiologia , Vírus/isolamento & purificação , Adolescente , Criança , Pré-Escolar , China/epidemiologia , Feminino , Humanos , Lactente , Recém-Nascido , Masculino , Técnicas de Diagnóstico Molecular , Nasofaringe/virologia , Prevalência , Estudos Prospectivos , Infecções Respiratórias/virologia , Estações do Ano , Topografia Médica , Viroses/virologia , Vírus/classificaçãoRESUMO
Acute respiratory tract infection is an important cause of morbidity and mortality with a worldwide disease burden. This study aimed to determine the prevalence and clinical characteristics of children with viral-induced acute respiratory tract infection, in Southern China. Nasopharyngeal aspirate samples from 1,980 pediatric patients with suspected acute respiratory tract infection, and 82 samples from healthy subject controls were collected for routine examination at the Second Affiliated Hospital of Shantou University Medical College, from October 2007 to August 2011. Specimens were tested by multiplex polymerase chain reaction (mPCR). At least one or more viruses were detected from 1,087 samples (54.9%). These included laboratory confirmations for 446 respiratory syncytial virus (RSV), 386 influenza virus A (FluA), 315 human rhinovirus (HRV), 135 human bocavirus (HBoV), 119 Parainfluenza virus 3 (PIV3), 82 Parainfluenza virus 1 (PIV1), 66 adenovirus (ADV), 53 WU polyomavirus (WUPyV), 52 human metapneumovirus (hMPV), and 29 influenza virus B (FluB) samples. Samples from healthy subjects were negative for any virus. Of the patients with positive specimens, 107 (9.8%) were admitted to pediatric intensive care unit (PICU). Co-infection with at least two of the viral pathogens under study was observed in 325 of the 1,980 patients (16.4% of the total number of cases). These findings may help in the diagnosis of viral infections of the respiratory tract in children, and help to consider current and potential therapeutic approaches for the treatment of acute respiratory tract infection, and further respiratory complications.
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Infecções Respiratórias/epidemiologia , Infecções Respiratórias/virologia , Viroses/epidemiologia , Viroses/virologia , Vírus/classificação , Vírus/isolamento & purificação , Criança , Pré-Escolar , China/epidemiologia , Feminino , Hospitais Universitários , Humanos , Lactente , Masculino , Nasofaringe/virologia , PrevalênciaRESUMO
OBJECTIVE: To study the characteristics of viral spectrum and clinical features of children in pediatric intensive care unit (PICU). METHOD: Nasopharyngeal aspirate specimens (NPA) from 349 patients(1 from each) and 130 cerebrospinal fluids (CSF) specimens were collected from children who were admitted to the PICU of Second Affiliated Hospital of Shantou University Medical College. Additional 87 NPA specimens were collected from healthy children for routine examination on the physical examination center, and the clinical data were collected. Multiplex PCR was applied to detect 16 kinds of viruses from NPA and CSF. Fluorescence quantitative PCR was applied to detect 13 viruses from CSF and to analyze the clinical data of positive cases. RESULT: There were 209 samples (59.9%) of the 349 NPA specimens were positive for viruses, which included 117 cases positive for human rhinovirus (HRV), 60 for respiratory syncytial virus (RSV), 20 for influenza virus A (Inf A), 10 for adenovirus (ADV), 6 for parainfluenza virus type 3(PIV-3), 6 for human Boca virus (HBoV), 5 for influenza virus C(Inf C), 4 for parainfluenza virus type 4(PIV-4), 4 for human coronavirus-HKU1/OC43, 3 for influenza virus B (Inf B), 3 for WU Polyomavirus (WUPyV), 2 parainfluenza virus type 1(PIV-1), 2 human metapneumovirus (HMPV) and 1 human coronavirus-NL63/229E. But none from 87 healthy controls were positive for any respiratory virus. Among the 130 CSF specimens, in 58 cases the diagnosis was viral encephalitis. There were 22 samples (37.9%) among the 58 CSF specimens positive for viruses, which included 14 enterovirus (EV), 3 human cytomegalovirus (HCMV), 2 mumps virus, 1 coxsackie virus A16 (Cox-A16), 1 herpes simplex virus (HSV) and 1 human rhinovirus (HRV). The total positive rate was 63.3% (221/349) . Co-infection by at least 2 viral pathogens under study was observed in 45 of the 349 patients (12.9% of the total number of cases, 20.4% of the positives cases). The commonest pathogens in co-infected samples were WUPyV (100%) and HMPV(100%). The positive rate of virus peaked in the first 6 months of life, the rate in boys were higher than in girls and the peak season was summer. The numbers of none serious cases in the virus positive group were less than those in the virus negative group while the numbers of extremely serious cases in the virus positive group were higher than in the virus negative group. CONCLUSION: Viral pathogen is a major cause of infectious disease in pediatric critical illnesses and virus infection may lead to severe illness.
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Unidades de Terapia Intensiva Pediátrica , Vírus de RNA/isolamento & purificação , Viroses/virologia , Doença Aguda , Distribuição por Idade , Criança , Pré-Escolar , Coinfecção/virologia , Encefalite Viral/epidemiologia , Encefalite Viral/virologia , Feminino , Humanos , Lactente , Vírus da Influenza A/isolamento & purificação , Masculino , Reação em Cadeia da Polimerase , Vírus Sinciciais Respiratórios/isolamento & purificação , Infecções Respiratórias/epidemiologia , Infecções Respiratórias/virologia , Rhinovirus/isolamento & purificação , Viroses/epidemiologiaRESUMO
OBJECTIVE: To investigate the prevalence and clinical features of human rhinovirus (HRV) infection in hospitalized children with acute respiratory (ARI) in eastern areas of Guangdong province from 2008 to 2010. METHODS: From Oct. 2008 through Sep. 2010, nasopharyngeal aspirates were collected prospectively, from hospitalized children with acute lower respiratory tract infection at the Second Hospital, affiliated to the Shantou University Medical College. Multiplex PCR was applied to detect ten kinds of viruses including HRV, RSV in the hospitalized children with respiratory tract infection. Clinical data on HRV-positive cases or RSV-positive cases were collected and analyzed. RESULTS: Among all the 1335 specimens, 124 were confirmed as HRV-positive cases (9.3%), with IVA-positive rate as the highest (25.1%), followed by RSV-positive rate (15.1%). HRV infection occurred sporadically around the year, with the highest HRV-positive rate seen in spring 2009 and autumn in 2010. Symptoms, signs, chest X-ray, leukocyte count and CRP count did not differ between patients with co-infection or single HRV infection. Clinical symptoms or signs were similar between those with single HRV infection or single RSV infection in children, but the single RSV infected children were more frequently seen with wheeze and cough. 28.4% of the single RSV infected children had bronchiolitis while 10.7% of single HRV infected children were seen (χ(2) = 0.281, P = 0.596). CONCLUSION: HRV was a relatively common cause for acute respiratory infections in the eastern areas of Guangdong province. The highest HRV-positive rate was slightly different in different years. Infants and young children were generally susceptible to rhinovirus infection. Bronchiolitis, wheeze and cough associated with HRV infection happened less than those caused by RSV.
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Infecções Respiratórias/epidemiologia , Infecções Respiratórias/virologia , Rhinovirus/isolamento & purificação , Criança , Criança Hospitalizada , Pré-Escolar , China/epidemiologia , Feminino , Humanos , Lactente , Masculino , Infecções por Vírus Respiratório Sincicial/epidemiologiaRESUMO
This study aimed at investigating the prevalence and clinical characteristics of children with respiratory infection by WU polyomavirus (WUPyV) in Southern China. Nasopharyngeal aspirate samples were collected from 771 children with acute respiratory tract infection admitted to hospital and 82 samples from healthy subjects for routine examination at the outpatient service at the Second Affiliated Hospital of Shantou University, Medical College from July 2008 to June 2009. WUPyV was detected by the polymerase chain reaction (PCR) and DNA sequencing. All WUPyV-positive specimens were characterized further for nine viruses causing common respiratory infections, including inï¬uenza A and B, respiratory syncytial virus (RSV), parainfluenza virus (PIV) 1 and 3, human metapneumovirus, human bocavirus, adenovirus, and rhinovirus by PCR or real time (RT)-PCR. Fifteen out of 771 specimens from patients with acute respiratory tract infection, but none from healthy subjects, were positive for WUPyV and the positivity rate was 2%. Patients with WUPyV infection were between 2 and 48 months of age, and nine of the patients were male while six female. Four out of 15 patients were co-infected with RSV, one with adenovirus or rhinovirus, respectively. Patients with WUPyV infection displayed predominantly cough, moderate fever, and wheezing, and were diagnosed with pneumonia (n = 8), bronchiolitis (n = 4), upper respiratory tract infections (n = 2) and bronchitis (n = 1). One patient developed encephalitis. Therefore, WUPyV infection can cause acute respiratory tract infection with atypical symptoms, including severe complications, in children.
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Infecções por Polyomavirus/epidemiologia , Infecções por Polyomavirus/patologia , Polyomavirus/isolamento & purificação , Doenças Respiratórias/epidemiologia , Doenças Respiratórias/patologia , Distribuição por Idade , Pré-Escolar , China/epidemiologia , Comorbidade , Feminino , Hospitalização , Humanos , Lactente , Masculino , Nasofaringe/virologia , Reação em Cadeia da Polimerase , Infecções por Polyomavirus/virologia , Prevalência , Doenças Respiratórias/virologia , Análise de Sequência de DNA , Distribuição por SexoRESUMO
Limb imaging spectrometer is an important new remote sensor for research and application. Signal-to-noise ratio (SNR) is one of the key parameters to quantitatively evaluate the image quality and radiometric performance of an imaging spectrometer. The estimation and testing of SNR are very important for developing an imaging spectrometer. From the perspectives of radiative transmission and energy conversion, the SNR model is proposed, and the SNR equation of dispersive-type limb imaging spectrometer is derived, and the SNR values under several observing conditions for an limb imaging spectrometer prototype developed are theoretically evaluated based on atmospheric radiative transfer code MODTRAN 4.0. The results show that the SNR of the prototype under typical viewing geometry is not less than 8. As experimental validation, SNR testing was performed using an internally illuminated integrating sphere, and the experimental results have proved the correctness of this theoretical model.
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OBJECTIVE: WU polyomavirus (WUPyV), a new member of the genus Polyomavirus in the family Polyomaviridae, has been found to be associated with respiratory tract infections recently. But the role of the WUPyV as agents of human disease remains uncertain. We sought to describe the detection and clinical characterization of WUPyV in acute respiratory tract infection in children. METHOD: From July 2008 through June 2009, nasopharyngeal aspirates were collected from 771 children who were hospitalized with acute respiratory tract infection in Second Affiliated Hospital of Shantou University Medical College, and from 82 asymptomatic children who visited the health checkup clinic. WUPyV was detected by using PCR technology and was identified by using DNA sequencing. All WUPyV-positive specimens were screened for 9 common viruses [influenza A and B, respiratory syncytial virus (RSV), parainfluenza virus (PIV) 1 and 3, human metapneumovirus, human bocavirus, adenovirus and rhinovirus] by using PCR or RT-PCR. The clinical data of WUPyV infection were collected and analyzed. RESULT: In this study, fifteen of the 771 tested specimens with acute respiratory tract infection were positive for WUPyV, the positive rate was 1.95% and all of the asymptomatic children who visited the health checkup clinic were negative. Of the 15 cases who were positive for the virus, the age range was 2 to 48 (mean 18.8) months, 9 (60%) were male and 6 (40%) were female. WUPyV was the sole virus detected in 9 specimens (60%) from patients with acute respiratory tract infection. WUPyV was associated with the co-infection with another respiratory virus in 6 of 15 (40%) cases, most frequently with RSV (n = 4), followed by adenovirus (n = 1) and rhinovirus (n = 1). The most common clinical findings in the patients with WUPyV were cough, fever and wheezing. The most frequent diagnoses were pneumonia (n = 8), bronchiolitis (n = 4), upper respiratory tract infections (n = 2) and bronchitis (n = 1). A severe case was complicated with viral encephalitis. CONCLUSION: WUPyV may be a respiratory pathogen because it was the sole virus detected in 9 specimens from patients with respiratory illness and all of the asymptomatic controls were negative. The most common clinical findings are cough and wheezing. Young children may be susceptible to infection with this virus and occasionally the infection with this virus may cause severe disease. More comprehensive and in-depth studies are required to prove the pathogenicity of these viruses.
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Infecções por Polyomavirus/virologia , Polyomavirus/isolamento & purificação , Infecções Respiratórias/virologia , Criança , Pré-Escolar , Feminino , Genes Virais , Humanos , Lactente , Recém-Nascido , Masculino , Reação em Cadeia da Polimerase , Polyomavirus/genética , Infecções por Polyomavirus/fisiopatologiaRESUMO
Recent studies have demonstrated that mesenchymal stem cells could differentiate into germ cells under appropriate conditions. We sought to determine whether human umbilical cord Wharton's jelly-derived mesenchymal stem cells (HUMSCs) could form germ cells in vitro. HUMSCs were induced to differentiate into germ cells in all-trans retinoic acid, testosterone and testicular-cell-conditioned medium prepared from newborn male mouse testes. HUMSCs formed "tadpole-like" cells after induction with different reagents and showed both mRNA and protein expression of germ-cell-specific markers Oct4 (POUF5), Ckit, CD49(f) (alpha6), Stella (DDPA3), and Vasa (DDX4). Our results may provide a new route for reproductive therapy involving HUMSCs and a novel in vitro model to investigate the molecular mechanisms that regulate the development of the mammalian germ lineage.
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Diferenciação Celular , Células Germinativas/citologia , Células-Tronco Mesenquimais/citologia , Animais , Biomarcadores/análise , Meios de Cultivo Condicionados/farmacologia , Humanos , Masculino , Células-Tronco Mesenquimais/efeitos dos fármacos , Camundongos , Testículo/citologia , Testículo/metabolismo , Testosterona/farmacologia , Tretinoína/farmacologia , Cordão UmbilicalRESUMO
OBJECTIVE: To investigate the viral pathogens of acute lower respiratory tract infection (ALRTI) in hospitalized children from East Guangdong Province of China and the relationship of the pathogens with age and seasons. METHODS: The nasopharyngeal aspirates samples obtained from 345 hospitalized children with ALRTI were investigated for respiratory syncytial virus (RSV), human bocavirus (HBoV), human metapneumovirus (hMPV), influenza virus types A and B, rhinovirus, parainfluenza virus types 1 and 3 and adenovirus by PCR. RESULTS: Viral pathogens were detected in 178 patients (51.6%). RSV was the most frequent (19.3%). Novel viruses hMPV (3.2%) and HBoV (3.2%) were found. A highest detection rate (61.9%) of virus was found between January to March. The infants aged 1 to 6 months showed a higher detection rate (71.3%) of virus than the other age groups. The detection rate of viral pathogens was 72.6% in children with bronchiolitis, followed by asthmatic bronchitis (70.0%) and bronchial pneumonia (44.6%). CONCLUSIONS: RSV remained the leading viral pathogens in children with ALRTI in East Guangdong of China. Novel viruses HBoV and hMPV were also important pathogens. The detection rate of viral pathogens was associated with seasonal changes and age. Different respiratory infectious diseases had different viral detection rates, with highest detection rate in bronchiolitis cases.