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1.
Anal Chem ; 93(33): 11634-11640, 2021 08 24.
Artigo em Inglês | MEDLINE | ID: mdl-34378382

RESUMO

Exploring the ratiometric fluorescence biosensing of DNA-templated biemissive silver nanoclusters (AgNCs) is significant in bioanalysis, yet the design of a stimuli-responsive DNA device is a challenge. Herein, using the anti-digoxin antibody (anti-Dig) with two identical binding sites as a model, a tweezer-like DNA architecture is assembled to populate fluorescent green- and red-AgNCs (g-AgNCs and r-AgNCs), aiming to produce a ratio signal via specific recognition of anti-Dig with two haptens (DigH). To this end, four DNA probes are programmed, including a reporter strand (RS) dually ended with a g-/r-AgNC template sequence, an enhancer strand (ES) tethering two same G-rich tails (G18), a capture strand (CS) labeled with DigH at two ends, and a help strand (HS). Initially, both g-AgNCs and r-AgNCs wrapped in the intact RS are nonfluorescent, whereas the base pairing between RS, ES, CS, and HS resulted in the construction of DNA mechanical tweezers with two symmetric arms hinged by a rigid "fulcrum", in which g-AgNCs are lighted up due to G18 proximity ("green-on"), and r-AgNCs away from G18 are still dark ("red-off"). When two DigHs in proximity recognize and bind anti-Dig, the conformation switch of these tweezers resultantly occurs, taking g-AgNCs away from G18 for "green-off" and bringing r-AgNCs close to G18 for "red-on". As such, the ratiometric fluorescence of r-AgNCs versus g-AgNCs is generated in response to anti-Dig, achieving reliable quantization with a limit of detection at the picomolar level. Based on the fast stimulated switch of unique DNA tweezers, our ratiometric strategy of dual-emitting AgNCs would provide a new avenue for a variety of bioassays.


Assuntos
Técnicas Biossensoriais , Nanopartículas Metálicas , Anticorpos , DNA , Fluorescência , Prata , Espectrometria de Fluorescência
2.
Pathogens ; 10(6)2021 May 31.
Artigo em Inglês | MEDLINE | ID: mdl-34072858

RESUMO

Cryptosporidium spp., Entamoeba histolytica, Giardia duodenalis, and Blastocystis sp. infections have been frequently reported as etiological agents for gastroenteritis, but also as common gut inhabitants in apparently healthy individuals. Between July 2016 and March 2017, stool samples (n = 507) were collected from randomly selected individuals (male/female ratio: 1.1, age range: 38-63 years) from two sentinel hospitals in Tengchong City Yunnan Province, China. Molecular (PCR and Sanger sequencing) methods were used to detect and genotype the investigated protist species. Carriage/infection rates were: Blastocystis sp. 9.5% (95% CI: 7.1-12.4%), G. duodenalis 2.2% (95% CI: 1.1-3.8%); and E. histolytica 2.0% (95% CI: 0.9-3.6%). Cryptosporidium spp. was not detected at all. Overall, 12.4% (95% CI: 9.7-15.6) of the participants harbored at least one enteric protist species. The most common coinfection was E. histolytica and Blastocystis sp. (1.0%; 95% CI: 0.3-2.2). Sequence analyses revealed that 90.9% (10/11) of the genotyped G. duodenalis isolates corresponded to the sub-assemblage AI. The remaining sequence (9.1%, 1/11) was identified as sub-assemblage BIV. Five different Blastocystis subtypes, including ST3 (43.7%, 21/48), ST1 (27.1%, 13/48), ST7 (18.8%, 9/48), ST4 (8.3%, 4/48), and ST2 (2.1%, 1/48) were identified. Statistical analyses confirmed that (i) the co-occurrence of protist infections was purely random, (ii) no associations were observed among the four protist species found, and (iii) neither their presence, individually or jointly, nor the patient's age was predictors for developing clinical symptoms associated with these infections. Overall, these protist mono- or coinfections are asymptomatic and do not follow any pattern.

3.
Infect Dis Poverty ; 10(1): 31, 2021 Mar 18.
Artigo em Inglês | MEDLINE | ID: mdl-33731163

RESUMO

BACKGROUND: The coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) has led to a significant number of mortalities worldwide. COVID-19 poses a serious threat to human life. The clinical manifestations of COVID-19 are diverse and severe and 20% of infected patients are reported to be in a critical condition. A loss in lung function and pulmonary fibrosis are the main manifestations of patients with the severe form of the disease. The lung function is affected, even after recovery, thereby greatly affecting the psychology and well-being of patients, and significantly reducing their quality of life. METHODS: Participants must meet the following simultaneous inclusion criteria: over 18 years of age, should have recovered from severe or critical COVID-19 cases, should exhibit pulmonary fibrosis after recovery, and should exhibit Qi-Yin deficiency syndrome as indicated in the system of traditional Chinese medicine (TCM). The eligible candidates will be randomized into treatment or control groups. The treatment group will receive modern medicine (pirfenidone) plus TCM whereas the control group will be administered modern medicine plus TCM placebo. The lung function index will be continuously surveyed and recorded. By comparing the treatment effect between the two groups, the study intend to explore whether TCM can improve the effectiveness of modern medicine in patients with pulmonary fibrosis arising as a sequelae after SARS-CoV-2 infection. DISCUSSION: Pulmonary fibrosis is one of fatal sequelae for some severe or critical COVID-19 cases, some studies reveal that pirfenidone lead to a delay in the decline of forced expiratory vital capacity, thereby reducing the mortality partly. Additionally, although TCM has been proven to be efficacious in treating pulmonary fibrosis, its role in treating pulmonary fibrosis related COVID-19 has not been explored. Hence, a multicenter, parallel-group, randomized controlled, interventional, prospective clinical trial has been designed and will be conducted to determine if a new comprehensive treatment for pulmonary fibrosis related to COVID-19 is feasible and if it can improve the quality of life of patients. TRIAL REGISTRATION: This multicenter, parallel-group, randomized controlled, interventional, prospective trial was registered at the Chinese Clinical Trial Registry (ChiCTR2000033284) on 26th May 2020 (prospective registered).


Assuntos
COVID-19/complicações , COVID-19/virologia , Fibrose Pulmonar/etiologia , Fibrose Pulmonar/terapia , SARS-CoV-2 , Antivirais/uso terapêutico , Terapia Combinada , Análise de Dados , Medicina Tradicional Chinesa , Fibrose Pulmonar/diagnóstico , Qualidade de Vida , Resultado do Tratamento
4.
Front Immunol ; 12: 616343, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33717108

RESUMO

Babesia microti is a protozoan that infects red blood cells. Babesiosis is becoming a new global threat impacting human health. Rhoptry neck proteins (RONs) are proteins located at the neck of the rhoptry and studies indicate that these proteins play an important role in the process of red blood cell invasion. In the present study, we report on the bioinformatic analysis, cloning, and recombinant gene expression of two truncated rhoptry neck proteins 2 (BmRON2), as well as their potential for incorporation in a candidate vaccine for babesiosis. Western blot and immunofluorescence antibody (IFA) assays were performed to detect the presence of specific antibodies against BmRON2 in infected mice and the localization of N-BmRON2 in B. microti parasites. In vitro experiments were carried out to investigate the role of BmRON2 proteins during the B. microti invasion process and in vivo experiments to investigate immunoprotection. Homologous sequence alignment and molecular phylogenetic analysis indicated that BmRON2 showed similarities with RON2 proteins of other Babesia species. We expressed the truncated N-terminal (33-336 aa, designated rN-BmRON2) and C-terminal (915-1171 aa, designated rC-BmRON2) fragments of the BmRON2 protein, with molecular weights of 70 and 29 kDa, respectively. Western blot assays showed that the native BmRON2 protein is approximately 170 kDa, and that rN-BmRON2 was recognized by serum of mice experimentally infected with B. microti. Immunofluorescence analysis indicated that the BmRON2 protein was located at the apical end of merozoites, at the opposite end of the nucleus. In vitro red blood cell invasion inhibition studies with B. microti rBmRON2 proteins showed that relative invasion rate of rN-BmRON2 and rC-BmRON2 group is 45 and 56%, respectively. Analysis of the host immune response after immunization and B. microti infection showed that both rN-BmRON2 and rC-BmRON2 enhanced the immune response, but that rN-BmRON2 conferred better protection than rC-BmRON2. In conclusion, our results indicate that truncated rhoptry neck protein 2, especially its N-terminal fragment (rN-BmRON2), plays an important role in the invasion of host red blood cells, confers immune protection, and shows good potential as a candidate vaccine against babesiosis.


Assuntos
Antígenos de Protozoários/imunologia , Babesia microti/imunologia , Babesiose/prevenção & controle , Interações Hospedeiro-Parasita/imunologia , Proteínas de Protozoários/imunologia , Vacinas Protozoárias/imunologia , Animais , Antígenos de Protozoários/genética , Babesia microti/genética , Modelos Animais de Doenças , Eritrócitos/imunologia , Eritrócitos/parasitologia , Imunofluorescência , Expressão Gênica , Imunização , Camundongos , Filogenia , Transporte Proteico , Proteínas de Protozoários/genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia
5.
Gene ; 766: 145153, 2021 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-32950633

RESUMO

AIM: Acute lung injury (ALI) is the mild form of acute respiratory distress syndrome (ARDS) which is a common lung disease with a high incidence and mortality rate. Recent studies manifested that some circular RNAs were associated with ALI. In this study, we aimed to uncover the effect of circular RNA circ_0054633 on ALI initiation and progression and proposed a new mechanism related to ALI. METHODS: The lipopolysaccharides (LPS)-induced acute lung injury model were build both in vivo of rat and in vitro of primary murine pulmonary microvascular endothelial cells (MPVECs). Hematoxylin and eosin (H&E) was employed to observe the tissue morphology and estimate the degree of lung damage. We used real-time quantitative polymerase chain reaction (RT-qPCR) to measure the expression level of circ_0054633. The expression levels of inflammatory cytokines IL-17A and tumor necrosis factor-α (TNF-α) were detected by ELISA. The effects of circ_0054633 on MPVECs proliferation and apoptosis were detected with the help of CCK-8 and apoptosis assay, separately. The expression level of NF-κB p65 protein was measured by Western blot. RESULTS: circ_0054633, IL-17A, TNF-α and NF-κB p65 were all overexpressed in LPS-treated rat and MPVECs, and LPS enhanced the proliferation and apoptosis of MPVECs. While circ_0054633 silencing reversed the above promotion effects of LPS on IL-17A, TNF-α expression and MPVECs proliferation and apoptosis. CONCLUSIONS: Quietness of circ_0054633 alleviated LPS-induced ALI via NF-κB signaling pathway, implicating circ_0054633 may be a potential biomarker for diagnose and therapy of ALI.


Assuntos
Lesão Pulmonar Aguda/metabolismo , Proliferação de Células/fisiologia , Células Endoteliais/metabolismo , Inflamação/metabolismo , NF-kappa B/metabolismo , RNA Circular/metabolismo , Lesão Pulmonar Aguda/induzido quimicamente , Animais , Apoptose/efeitos dos fármacos , Apoptose/fisiologia , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Inflamação/induzido quimicamente , Interleucina-17/metabolismo , Lipopolissacarídeos/farmacologia , Pulmão/efeitos dos fármacos , Pulmão/metabolismo , Masculino , Camundongos , Ratos , Ratos Sprague-Dawley , Fator de Transcrição RelA/metabolismo , Fator de Necrose Tumoral alfa/metabolismo
6.
Oncol Rep ; 41(5): 3100-3110, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30976815

RESUMO

The pleiotropic effects of hyperthermia on cancer cells have been well documented, and microwave hyperthermia (MWHT) has been widely applied for multifarious cancer treatment. However, the mechanisms underlying the anticancer effect of MWHT combined with gemcitabine (GEM) remain poorly understood. The aim of the present study was to investigate the role of autophagy in the thermo­chemotherapy of human squamous cell lung carcinoma cells. It was observed that MWHT combined with GEM potently suppressed the viability of NCI­H2170 and NCI­H1703 cells, and induced G0/G1 cell cycle arrest. Notably, MWHT with GEM induced autophagy, as indicated by the formation of autophagic vacuoles, downregulation of p62 and upregulation of light chain 3­II. It was further demonstrated that the autophagy was due to the production of reactive oxygen species (ROS), whereas N­acetyl cysteine, an ROS scavenger, attenuated the level of autophagy. However, when the autophagy inhibitor 3­methyladenine was used, there was no significant change in the production of ROS. Furthermore, it was observed that MWHT combined with GEM downregulated the protein expression levels of phosphoinositide 3­kinase (PI3K), phosphorylated (p)­PI3K, protein kinase B (AKT), p­AKT, mammalian target of rapamycin (mTOR), p­mTOR, phosphorylated S6 (pS6) and p70 S6 kinase, which are associated with autophagy. In addition, the results demonstrated that ROS served as an upstream mediator of PI3K/AKT/mTOR signaling. In light of these findings, the present study provides original insights into the molecular mechanisms underlying the cell death induced by MWHT combined with GEM, and this may be a promising approach for the treatment of human squamous cell lung carcinoma.


Assuntos
Autofagia/efeitos da radiação , Carcinoma Pulmonar de Células não Pequenas/terapia , Desoxicitidina/análogos & derivados , Hipertermia Induzida/métodos , Neoplasias Pulmonares/terapia , Adenina/análogos & derivados , Adenina/farmacologia , Autofagia/efeitos dos fármacos , Carcinoma Pulmonar de Células não Pequenas/patologia , Linhagem Celular Tumoral , Terapia Combinada/métodos , Desoxicitidina/farmacologia , Desoxicitidina/uso terapêutico , Resistencia a Medicamentos Antineoplásicos/efeitos da radiação , Humanos , Neoplasias Pulmonares/patologia , Micro-Ondas/uso terapêutico , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/efeitos da radiação , Serina-Treonina Quinases TOR/metabolismo
7.
Infect Dis Poverty ; 7(1): 100, 2018 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-30318019

RESUMO

BACKGROUND: Angiostrongyliasis is a food-borne parasitic zoonosis. Human infection is caused by infection with the third-stage larvae of Angiostrongylus cantonensis. The life cycle of A. cantonensis involves rodents as definitive hosts and molluscs as intermediate hosts. This study aims to investigate on the infection status and characteristics of spatial distribution of these hosts, which are key components in the strategy for the prevention and control of angiostrongyliasis. METHODS: Three villages from Nanao Island, Guangdong Province, China, were chosen as study area by stratified random sampling. The density and natural infection of Pomacea canaliculata and various rat species were surveyed every three months from December 2015 to September 2016, with spatial correlations of the positive P. canaliculata and the infection rates analysed by ArcGIS, scan statistics, ordinary least squares (OLS) and geographically weighted regression (GWR) models. RESULTS: A total of 2192 P. canaliculata specimens were collected from the field, of which 1190 were randomly chosen to be examined for third-stage larvae of A. cantonensis. Seventy-two Angiostrongylus-infected snails were found, which represents a larval infection rate of 6.1% (72/1190). In total, 110 rats including 85 Rattus norvegicus, 10 R. flavipectus, one R. losea and 14 Suncus murinus were captured, and 32 individuals were positive (for adult worms), representing an infection rate of 29.1% of the definitive hosts (32/110). Worms were only found in R. norvegicus and R. flavipectus, representing a prevalence of 36.5% (31/85) and 10% (1/10), respectively in these species, but none in R. losea and S. murinus, despite testing as many as 32 of the latter species. Statistically, spatial correlation and spatial clusters in the spatial distribution of positive P. canaliculata and positive rats existed. Most of the spatial variability of the host infection rates came from spatial autocorrelation. Nine spatial clusters with respect to positive P. canaliculata were identified, but only two correlated to infection rates. The results show that corrected Akaike information criterion, R2, R2 adjusted and σ2 in the GWR model were superior to those in the OLS model. CONCLUSIONS: P. canaliculata and rats were widely distributed in Nanao Island and positive infection has also been found in the hosts, demonstrating that there was a risk of angiostrongyliasis in this region of China. The distribution of positive P. canaliculata and rats exhibited spatial correlation, and the GWR model had advantage over the OLS model in the spatial analysis of hosts of A. cantonensis.


Assuntos
Angiostrongylus cantonensis , Especificidade de Hospedeiro , Interações Hospedeiro-Parasita , Animais , China/epidemiologia , Geografia , Humanos , Ratos , Caramujos/parasitologia , Análise Espacial , Infecções por Strongylida/epidemiologia , Infecções por Strongylida/parasitologia , Infecções por Strongylida/transmissão , Zoonoses
8.
Zhongguo Zhong Yao Za Zhi ; 43(8): 1654-1661, 2018 Apr.
Artigo em Chinês | MEDLINE | ID: mdl-29751713

RESUMO

The chemical constituents from the n-butanol fraction of the 70% ethanol extract of Datura metel roots were separated by silica gel and ODS chromatogram columns as well as preparative HPLC. On the basis of spectral data analysis, their structures were elucidated. Twenty-one compounds were obtained and their structures were identified as citroside A (1), coniferin (2), paeoniflorin (3), (6R,7E,9R)-9-hydroxy-4,7-megastigmadien-3-one 9-O-[α-L-arabin-opyranosyl-(l→6)-ß-D-glucopyranoside] (4), (1R,7R,10R,11R)-12-hydroxyl anhuienosol (5), kaurane acid glycoside A (6), ent-2-oxo-15,16-dihydroxypimar-8(14)-en-16-O-ß-glucopyranoside (7), ginsenoside Rg1(8), ginsenoside Re (9), notoginsenosides R1(10), N-butyl-O-ß-D-fructofuranoside (11), salidroside (12), hexyl ß-sophoroside (13), 2,6-dimethoxy-4-hydroxyphenol 1-glucoside (14), benzyl-O-ß-D-xylopyranoxyl(1→6)-ß-D-glucopyranoside (15), (Z)-3-hexenyl-O-α-L-arabinopyranosyl-(1→6)-ß-D-glucopyranoside (16), N-[2-(3,4-dihydro-xyphenyl)-2-hydroxyethyl]-3-(4-methoxyphenyl) prop-2-enamide (17), cannabisin D (18), cannabisin E (19), melongenamide B (20), paprazine (21). Compounds 2-17 and 20-21 were isolated from the Solanaceae family for the first time.


Assuntos
Datura metel , Medicamentos de Ervas Chinesas , Etanol , Glicosídeos , Raízes de Plantas
9.
Acta Trop ; 185: 371-379, 2018 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-29559329

RESUMO

Babesiosis has become a new global threat impacting human health, and most human babesiosis cases are caused by Babesia microti. Until now few antigens of B. microti have been described which can be used for the diagnosis of human babesiosis. In the present study, we report on the bioinformatic analysis, cloning and expression of the sequence encoding the B. microti seroreactive antigen 5-1-1 to investigate its potential incorporation in serologic diagnostic tools for babesiosis. Bioinformatic analysis and recombinant gene expression were performed to molecularly characterize seroreactive antigen 5-1-1. Enhanced chemiluminescence (ECL)-Western blot methods were used to detect specific antibodies in infected mice. Immunofluorescence antibody assays (IFA) were performed to detect the localization of BmSA5-1-1 in B. microti parasites. ELISA and immunochromatographic (ICT) tests were developed using recombinant BmSA5-1-1 to evaluate its potential use in rapid detection methods for B. microti antibodies and for the diagnosis of babesiosis. A recombinant expression plasmid was constructed by inserting the target gene fragment in the pET28a vector after double digestion with BamHI and XhoI restriction enzymes. The recombinant BmSA5-1-1 protein was expressed in Escherichia coli (rBmSA5-1-1) and purified by means of Ni-nitrilotriacetic acid (NTA) agarose columns. Polyclonal antibodies were generated against rBmSA5-1-1. Based on indirect immunofluorescence assay results, BmSA5-1-1 appeared to localize on the surface of B. microti. ELISA tests using the rBmSA5-1-1 antigen detected specific antibodies from infected mice as early as 4 days post-infection. Our results indicate that the two methods we developed can detect specific antibodies in mice at different stages of infection with sensitivities of 100% (rBmSA5-1-1 ELISA) and 90% (ICT). The specificity of the two methods was 100%. Sera of patients suffering from other closely related parasitic diseases, such as malaria and toxoplasmosis, produced negative results. In conclusion, seroreactive antigen 5-1-1, a member of the BMN1 protein family, is expressed on the outer surface of B. microti and is a promising candidate antigen for the early diagnosis of babesiosis. rBmSA5-1-1 ELISA and ICT methods show good potential for detecting specific antibodies in mice at different stages of infection.


Assuntos
Anticorpos Antiprotozoários/sangue , Antígenos de Protozoários/genética , Babesia microti/imunologia , Babesiose/diagnóstico , Técnica Indireta de Fluorescência para Anticorpo/métodos , Animais , Antígenos de Protozoários/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Proteínas Recombinantes/imunologia
10.
Neurosci Lett ; 651: 134-139, 2017 06 09.
Artigo em Inglês | MEDLINE | ID: mdl-28479104

RESUMO

Migraine is a debilitating disorder characterized by recurrent headache arising from neurovascular dysfunction. Despite recent progress in migraine research, the exact mechanisms underpinning migraine are poorly understood. Furthermore, it is difficult to develop an animal model of migraine that resembles all symptoms of patients. In this study, we established a novel animal model of migraine induced by epidural injection of calcitonin gene-related peptide (CGRP), and examined climbing hutch behavior, facial-grooming behavior, body-grooming behavior, freezing behavior, resting behavior, and ipsilateral hindpaw facial grooming behavior of rats following CGRP injection. CGRP significantly reduced climbing hutch behavior, and face-grooming behavior, and increased immobile behavior. We also found that the P15 and P85 percentile range of behavioral data exhibited a high positive rate (83.3%) for establishing the model with less false positive rate. Our results verified that the rat model of migraine induced by CGRP featured many behaviors of migraine patients demonstrated during migraine attacks. Our findings suggest that this new model can be a useful tool for understanding the pathophysiology of migraine and studying novel therapeutic strategies for the treatment of migraine.


Assuntos
Comportamento Animal/efeitos dos fármacos , Peptídeo Relacionado com Gene de Calcitonina/administração & dosagem , Modelos Animais de Doenças , Transtornos de Enxaqueca/psicologia , Animais , Asseio Animal/efeitos dos fármacos , Masculino , Transtornos de Enxaqueca/induzido quimicamente , Ratos Sprague-Dawley
11.
Gut Pathog ; 8: 58, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27891182

RESUMO

BACKGROUND: Acute diarrhea is one of the major public health problems worldwide. Most of studies on acute diarrhea have been made on infants aged below 5 years and few efforts have been made to identify the etiological agents of acute diarrhea in people over five, especially in China. METHODS: 271 diarrhea cases and 149 healthy controls over 5 years were recruited from four participating hospitals between June 2014 and July 2015. Each stool specimen was collected to detect a series of enteric pathogens, involving five viruses (Rotavirus group A, RVA; Norovirus, NoV; Sapovirus, SaV; Astrovirus, As; and Adenovirus, Ad), seven bacteria (diarrheagenic Escherichia coli, DEC; non-typhoidal Salmonella, NTS; Shigella spp.; Vibrio cholera; Vibrio parahaemolyticus; Aeromonas spp.; and Plesiomonas spp.) and three protozoa (Cryptosporidium spp., Giardia lamblia, G. lamblia, and Blastocystis hominis, B. hominis). Standard microbiological and molecular methods were applied to detect these pathogens. Data was analyzed using Chi square, Fisher-exact tests and logistic regressions. RESULTS: The prevalence of at least one enteric pathogen was detected in 29.2% (79/271) acute diarrhea cases and in 12.1% (18/149) in healthy controls (p < 0.0001). Enteric viral infections (14.4%) were the most common in patients suffering from acute diarrhea, followed by bacteria (13.7%) and intestinal protozoa (4.8%). DEC (12.5%) was the most common causative agent in diarrhea cases, followed by NoV GII (10.0%), RVA (7.4%) and B. hominis (4.8%). The prevalence of co-infection was statistically higher (p = 0.0059) in the case group (7.7%) than in the healthy control (1.3%). RVA-NoV GII (3.0%) was the most common co-infection in symptomatic cases. CONCLUSIONS: DEC was the most predominant pathogen in diarrhea cases, but it was largely overlooked because the lack of laboratory capacities. Because of the high prevalence of co-infections, it is recommended the urgent development of alternative laboratory methods to assess polymicrobial infections. Such methodological improvements will result in a better prevention and treatment strategies to control diarrhea illness in China.

12.
Zhonghua Wei Zhong Bing Ji Jiu Yi Xue ; 25(8): 475-8, 2013 Aug.
Artigo em Chinês | MEDLINE | ID: mdl-24021043

RESUMO

OBJECTIVE: To observe the effect of nystatin on incidence of invasive fungal infections (IFI) and the prognosis of mechanically ventilated critically ill patients. METHODS: A prospective study was conducted. Critical ill patients admitted to Department of Critical Care Medicine of Jiangxi Provincial People's Hospital from May 1st, 2012 to April 30th, 2013 needing mechanical ventilation were enrolled. The patients were randomly divided into two groups by envelope method. Patients in the nystatin group were administered nystatin 1000 kU three times a day via the gastric tube; and patients in the control group were given gastrointestinal prokinetic drug as placebo. The specimens were collected every 3 days throughout the ICU stay (T0, T3, T6, T9), the strain distribution was observed, and the corrected colonization index (CCI) of all patients were calculated. The incidence of candidemia and 28-day mortality as well as the duration of stay in ICU and hospital were also recorded. RESULTS: A total of 874 strains were isolated from 124 patients, of which Candida albicans accounted for 57.6% (503/874). The most frequently colonized body sites were oropharyngeal site,account for 35.6% (311/874). The CCI of the nystatin group were lower than those of the control group at T6 and T9 [T6: 0.19±0.10 vs. 0.39±0.15, T9: 0.00 (0.10) vs. 0.45 (0.30), all P<0.05]. The incidence of candidemia in the nystatin group was slightly lower than that in control group [0.5% (3/60) vs. 7.8% (5/64), P>0.05]. The mortality in the nystatin group was lower than that in control group [18.3% (11/60) vs. 34.4% (22/64), P<0.05]. ICU day in the nystatin group was shorter than that in the control group (days: 9.56±3.47 vs. 11.89±6.32, P<0.05). However,hospital day was similar in the two groups (days: 18.35±7.42 vs. 20.58±8.77, P>0.05). CONCLUSIONS: Nystatin might reduce the colonization of Candida albicans and was associated with shorter ICU day.


Assuntos
Micoses/prevenção & controle , Nistatina/uso terapêutico , Respiração Artificial/efeitos adversos , Adulto , Idoso , Candida albicans/isolamento & purificação , Candidemia/prevenção & controle , Feminino , Humanos , Unidades de Terapia Intensiva , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos
13.
Rheumatol Int ; 32(3): 767-71, 2012 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-21193990

RESUMO

The minor allele of the non-synonymous single nucleotide polymorphism (SNP) +1858C>T within the PTPN22 gene has now been unequivocally confirmed as conferring susceptibility to RA in population from Europe and America, but not in population from Asia. The aim of this study was to jointly address and integrate these separate findings to further elucidate the association between the PTPN22 gene and RA in Chinese Hans of Guangdong province. Four hundred and ninety-four cases with RA and 496 healthy controls were randomly selected, their SNPs at position -1123G>C (rs2488457), +1858C>T (rs2476601), +788G>A (rs33996649), and rs1310182 were genotyped using PCR-RFLP, followed by agarose gel electrophoresis. +1858C>T (rs2476601) and +788G>A (rs33996649) are not polymorphic in Chinese Hans. Meanwhile, our result reveals that the degree of association between the promoter polymorphism, -1123G>C and RA, was analogous to that observed in Japanese reports (odds ratio [OR] = 1.517, 95% CI = [1.154-1.995], P = 0.003). Expression study also indicated a tendency for association between -1123G>C and PTPN22 gene expression. Our study underpins that the promoter polymorphism, -1123G/C, may be a causal SNP for RA in Asian.


Assuntos
Artrite Reumatoide/genética , Grupo com Ancestrais do Continente Asiático/genética , Predisposição Genética para Doença , Polimorfismo de Nucleotídeo Único , Proteína Tirosina Fosfatase não Receptora Tipo 22/genética , Adulto , China , Feminino , Expressão Gênica , Genótipo , Humanos , Masculino , Regiões Promotoras Genéticas
14.
Environ Microbiol Rep ; 3(5): 603-12, 2011 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-23761341

RESUMO

A fish pathogen Edwardsiella tarda LTB-4 produced various indole alkaloids, including indole, 2-(1H-indol-3-yl)ethanol, 4-di(1H-indol-3-yl)methylphenol, tri(1H-indol-3-yl)methane and 2-[2,2-bis(1H-indol-3-yl)]ethylphenylamine. Indole was the most abundant among these indole alkaloids. E. tarda LTB-4 produced indoles during its whole growth phase and maintained a high level (around 35.5 µM) during the stationary phase. The relevant tryptophanase (TnaA) gene tnaA was cloned from LTB-4 and conditionally expressed in Escherichia coli; the recombinant TnaA catalysed L-tryptophan to indole. A tnaA in-frame deletion mutant ΔtnaA was constructed through double cross-over allelic exchange by means of the suicide vector pRE118; deletion of tnaA caused some phenotypic changes including decreased swarming and twitching motility, lipopolysaccharide production and multiple antibiotic resistances. Also, subtherapeutic doses of chloromycetin, carbenicillin and tetracyline could cause the decrease of bacterial growth, but greatly induce the production of indole by E. tarda. Most importantly, attenuated virulence of the ΔtnaA mutant to zebra fish by increasing the LD50 for about 55-fold indicated that TnaA involved in the virulence of E. tarda.

15.
Otolaryngol Head Neck Surg ; 143(6): 765-71, 2010 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-21109075

RESUMO

OBJECTIVE: Pepsin detection in throat sputum has been posited as a reliable biological marker of laryngopharyngeal reflux disease (LPRD). This study was designed to further correlate pepsin concentration with symptoms and signs of LPRD. STUDY DESIGN: Cross-sectional study. SETTING: Nanfang Hospital of Southern Medical University. SUBJECTS AND METHODS: Fifty-six laryngitis patients were divided into a reflux laryngitis group and a chronic laryngitis group based on the reflux symptom index (RSI), reflux finding score (RFS), and proton pump inhibitor treatment for two weeks. Oral and hypopharyngeal secretions from the study patients and from 15 healthy subjects were collected. Thirty-six obstructive sleep apnea (OSA) patients were divided into a mild-moderate group and a severe group by the apnea-hypopnea index (AHI). Bedtime and first-morning oral secretions were collected. Enzyme-linked immunosorbent assay was used to measure the pepsin concentration. RESULTS: In laryngitis patients, the total score of RSI and RFS (P < 0.05), and the symptoms, including clearing throat often, coughing, and sensing a lump in the throat (P < 0.006), were more severe in the pepsin-positive group. No significant differences were found between the oral and hypopharyngeal secretions. In OSA, pepsin levels in the first-morning oral secretions were correlated with AHI, mean SaO(2), and mini SaO(2) (P < 0.01). However, RSIs were not significantly correlated with these indicators. CONCLUSION: Higher levels of pepsin in sputum were associated with higher RSI and RFS in cases of laryngitis. There was no relationship between pepsin levels and RSI in cases of OSA. There were no differences of pepsin concentration in sputum collection methods or in collection timing.


Assuntos
Laringite/diagnóstico , Refluxo Laringofaríngeo/diagnóstico , Pepsina A/análise , Escarro/química , Adulto , Biomarcadores/análise , Estudos Transversais , Feminino , Humanos , Laringite/etiologia , Refluxo Laringofaríngeo/complicações , Masculino , Pessoa de Meia-Idade , Faringe
16.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi ; 24(9): 857-60, 2008 Sep.
Artigo em Chinês | MEDLINE | ID: mdl-18782515

RESUMO

AIM: To silence RelB gene in mouse bone-marrow derived dendritic cells (DC) utilizing lentiviral vector, a novel tolerogenic dendritic cell with a relatively low expression level RelB was constructed and a new way to treat and prevent autoimmune diseases was explored. METHODS: Interferential targeting sequence R5 of RelB in mice was designed, synthesized and cloned into lentiviral vectors. Together with viral packaging materials were co-cultured in 293FT cell line to package lentiviral vector. Supernatant fluids were harvested, then virus titer detected. Mouse bone marrow derived DCs were infected by lentivirus particle. RelB gene expression level was detected by RT-PCR and immunofluorescence staining and analyzed by software of geo pro. There are three experiment control groups including immature DC, mature DC and DC infected by a negative independent control of T6. RESULTS: A similar RelB expression was detected by RT-PCR and immunofluorescence staining assay between DC infected virus R5 and immature DC, but was lower than that of mature DC. Significant difference in statistics P < 0.05. A similar RelB expression was detected by RT-PCR and immunofluorescence staining approaches between DC infected virus T6 and mature DC, but was higher than that of immature DC. Significant difference in statistics P < 0.05. CONCLUSION: RelB gene expressed by mouse bone marrow derived DC was silenced by Lentivirus vector effectively. The lentivirus vector with a low immunogenicity can be used to immunotherapy in vivo and overcome difficult transfection problem of primary DC. A new viral vector of DC immunotherapy can be obtained.


Assuntos
Células da Medula Óssea/metabolismo , Células Dendríticas/metabolismo , Inativação Gênica , Marcação de Genes/métodos , Fator de Transcrição RelB/genética , Animais , Células da Medula Óssea/citologia , Células Cultivadas , Células Dendríticas/virologia , Vetores Genéticos/genética , Vetores Genéticos/metabolismo , Humanos , Lentivirus/genética , Lentivirus/metabolismo , Camundongos , RNA Interferente Pequeno/genética , RNA Interferente Pequeno/metabolismo , Fator de Transcrição RelB/metabolismo
17.
Nan Fang Yi Ke Da Xue Xue Bao ; 28(8): 1382-6, 2008 Aug.
Artigo em Chinês | MEDLINE | ID: mdl-18753066

RESUMO

OBJECTIVE: To establish the genotype-specific targets plasmids and engineered E.coli strains of botulinum neurotoxins (BoNT) types B and E based on reverse genetics. METHODS: The gene sequences of BoNT were obtained from GenBank and analyzed using DNAMAN, Lasergene, Vector NTI and BLAST. Two target fragments of BoNT/B and BoNT/E were anchored and then synthesized as 5 and 10 short DNA single strands, respectively. The full-length target sequences were amplified by overlapping PCR and subcloned into pMD 18-T vector, and the recombinant plasmids were identified by restriction enzyme digestion and sequencing. RESULTS: Sixty full-length sequences of 4 types of BoNT, namely types A, B, E, and F, were available in GenBank. Two target fragments, BoNT/B of 215 bp and BoNT/E of 360 bp, and their specific primer pairs were anchored after sequence analysis. pMD 18-T-BoNT/B and pMD 18-T-BoNT/E containing these two target sequences were confirmed. CONCLUSION: The engineered plasmids and E.coli stains containing the genotype-specific target fragments of BoNT/B and BoNT/E have been constructed successfully.


Assuntos
Toxinas Botulínicas/genética , Clostridium botulinum/isolamento & purificação , Marcação de Genes , Sequência de Bases , Toxinas Botulínicas Tipo A , Clostridium botulinum/genética , Genótipo , Dados de Sequência Molecular , Reação em Cadeia da Polimerase/métodos , Análise de Sequência de DNA
18.
Nan Fang Yi Ke Da Xue Xue Bao ; 28(5): 866-9, 2008 May.
Artigo em Chinês | MEDLINE | ID: mdl-18504223

RESUMO

OBJECTIVE: To detect the changes in serum N-terminal pro-brain natriuretic peptide (NT-proBNP) levels in patients with cardiovascular diseases and explore its clinical significance. METHODS: Serum NT-proBNP concentrations were measured by electrochemiluminescent immunoassay (ECLIA) in 460 patients with cardiovascular diseases and in 50 normal controls, and echocardiographic examination was performed to determine the left ventricular ejection function (LVEF). Analysis of NT-proBNP was performed for its correlation to New York Heart Association (NYHA) functional classifications LVEF, and risk factors of cardiovascular diseases. RESULTS: The serum LgNT-proBNP concentrations was 3.74 in patients with cardiovascular diseases, significantly higher than that of normal controls (1.42, P<0.001). NT-proBNP concentrations also varied significantly among patients with different cardiovascular diseases as shown by one-way ANOVA analysis (F=17.761, P<0.001). The NT-proBNP levels increased with the severity of heart failure according to NYHA functional classifications (P<0.001), and varied significantly in patients suffering different cardiovascular diseases with the same NYHA functional class. Multivariable regression analysis indicated there were significant correlations of NT-proBNP levels with the patients' age (r=0.152, P<0.001), NYHA functional classifications (r=0.725, P<0.001), LVEF (r=-0.634, P<0.001), and clinica outcomes (r=-0.581, P<0.001). Logistic regression analysis identified NT-proBNP level as a strong indicator for cardiovascular events (HR=2.763, P<0.01) with close correlation to the treatment results. CONCLUSIONS: Serum NT-proBNP level varies significantly with the severity of heart failure and can be indicative of the patients' cardiac function in close correlation to the clinical prognosis, but its value for diagnostic stratification of cardiovascular diseases awaits further investigation.


Assuntos
Doença das Coronárias/sangue , Insuficiência Cardíaca/sangue , Peptídeo Natriurético Encefálico/sangue , Fragmentos de Peptídeos/sangue , Volume Sistólico/fisiologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Doença das Coronárias/fisiopatologia , Feminino , Insuficiência Cardíaca/fisiopatologia , Humanos , Imunoensaio/métodos , Masculino , Pessoa de Meia-Idade , Função Ventricular , Adulto Jovem
19.
Nan Fang Yi Ke Da Xue Xue Bao ; 27(8): 1277-9, 2007 Aug.
Artigo em Chinês | MEDLINE | ID: mdl-17715048

RESUMO

OBJECTIVE: To explore the diagnostic value of serum anti-cyclic citrullinated peptide (Anti-CCP) antibodies in patients with rheumatoid arthritis (RA). METHODS: Anti-CCP antibodies were detected in the serum samples of 120 RA patients, 71 non-RA patients with various rheumatic diseases, and 50 normal controls by enzyme-linked immunosorbent assay (ELISA) using domestic and imported commercial detection kits. Rheumatoid factors (RF) were assayed by immune-nephelometry. The correlation between Anti-CCP and RF in RA diagnosis was analyzed by calculating the area under curve of the receiver operating characteristic (ROC) curve. RESULTS: The positive rates for Anti-CCP, detected using both domestic and imported kits, were 61.7% (74/120) and 69.2% (83/120) in RA group, significantly higher than those in the non-RA group (9.9%, 7/71 and 7.0%, 5/71) and normal control group (both 0, P<0.001). The sensitivities for Anti-CCP and RF were 69.2% and 64.2%, and the specificities were 92.9% and 67.6%, respectively. The positive predictive value was 94.3% for Anti-CCP and 77.0% for RF, whereas the negative predictive value was 64.1% for Anti-CCP and 52.7% for RF. The likelihood ratio (LR) was 9.82 for anti-CCP and 1.98 for RF. The area under curve of ROC for Anti-CCP was 0.829 and 0.740 for RF. Anti-CCP antibodies had greater diagnostic value than RF in RA diagnosis, and Anti-CCP showed significant correlation with RF (r=0.29, P=0.001). CONCLUSION: Anti-CCP antibodies are an excellent serological marker for RA, which shows high diagnostic specificity at early stage, and can increase its diagnostic value when combined with RF detection, but the role of Anti-CCP in the occurrence and prognosis of RA remains to be further investigated.


Assuntos
Anticorpos/sangue , Anticorpos/imunologia , Artrite Reumatoide/sangue , Artrite Reumatoide/diagnóstico , Peptídeos Cíclicos/imunologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Criança , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Curva ROC , Fator Reumatoide/sangue , Adulto Jovem
20.
Di Yi Jun Yi Da Xue Xue Bao ; 24(7): 818-20, 2004 Jul.
Artigo em Chinês | MEDLINE | ID: mdl-15257913

RESUMO

OBJECTIVE: To assess the prognostic value of changes of peripheral blood T cell subsets after thermoradiotherapy for nasopharyngeal carcinoma (NPC). METHODS: Peripheral blood T cell subsets in 20 normal subjects (control group), 30 NPC patients undergoing thermoradiotherapy, and 20 NPC patients undergoing radiotherapy were detected by flow cytometry. RESULTS: The percentages of CD3+CD4+ and CD8+CD28+ cells were decreased and the percentages of CD3+CD8+ and CD8+CD28- cells increased as compared with the measurements in normal persons. One month after thermoradiotherapy, the percentages of CD3+CD4+ and CD8+CD28+ cells further decreased and the percentages of CD3+CD8+ and CD8+CD28- cells further increased, which continued to worsen 3 months after the treatment and appeared to be related to the survival of the patients. CONCLUSION: T cell subsets of NPC patients are abnormal and their immune functions depressed in NPC patients within a long period after thermoradiotherapy. CD8+CD28+ and CD8+CD28- T cell subsets can be significant for prognostic assessment in these patients after thermoradiotherapy.


Assuntos
Hipertermia Induzida , Neoplasias Nasofaríngeas/terapia , Subpopulações de Linfócitos T/imunologia , Adulto , Antígenos CD28/análise , Antígenos CD8/análise , Terapia Combinada , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Neoplasias Nasofaríngeas/imunologia , Prognóstico
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