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1.
Anal Chem ; 93(14): 5839-5848, 2021 04 13.
Artigo em Inglês | MEDLINE | ID: mdl-33797890

RESUMO

The multiple-metal-nanoparticle tagging strategy has generally been applied to the multiplexed detection of multiple analytes of interest such as microRNAs (miRNAs). Herein, it was used for the first time to improve both the specificity and sensitivity of a novel mass spectroscopic platform for miRNA detection. The mass spectroscopic platform was developed through the integration of the ligation reaction, hybridization chain reaction amplification, multiple-metal-nanoparticle tagging, and inductively coupled plasma mass spectrometry. The high specificity resulted from the adoption of the ligation reaction is further enhanced by the multiple-metal-nanoparticle tagging strategy. The combination of hybridization chain reaction amplification and metal nanoparticle tagging endows the proposed platform with the feature of high sensitivity. The proposed mass spectrometric platform achieved quite satisfactory quantitative results for Let-7a in real-world cell line samples with accuracy comparable to that of the real-time quantitative reverse-transcriptase polymerase chain reaction method. Its limit of detection and limit of quantification for Let-7a were experimentally determined to be about 0.5 and 10 fM, respectively. Furthermore, due to the unique way of utilizing the multiple-metal-nanoparticle tagging strategy, the proposed platform can unambiguously discriminate between the target miRNA and nontarget ones with single-nucleotide polymorphisms based on their response patterns defined by the relative mass spectral intensities among the multiple tagged metal elements and can also provide location information of the mismatched bases. Its unique advantages over conventional miRNA detection methods make the proposed platform a promising and alternative tool in the fields of clinical diagnosis and biomedical research.


Assuntos
Nanopartículas Metálicas , MicroRNAs , Limite de Detecção , Espectrometria de Massas , MicroRNAs/genética , Hibridização de Ácido Nucleico
2.
Talanta ; 224: 121848, 2021 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-33379064

RESUMO

A facile MALDI-TOF mass spectrometric platform for quantitative analysis of protein biomarkers was developed based on magnetic ZnFe2O4 nanoparticles and mass tagging signal amplification. In this platform, magnetic ZnFe2O4 nanoparticles functionalized with an aptamer of the biomarker of interest was used to magnetically separate silica nanoparticles modified with another aptamer of the target biomarker and a barcoding peptide from solution phase in the presence of the biomarker of interest. After the silica nanoparticles were dissolved by KHF2, the released barcoding peptide was detected by MALDI-TOF mass spectrometry with magnetic ZnFe2O4 nanoparticles used as assisting matrix of laser desorption ionization. Since the mass spectral intensity of the barcoding peptide is directly related to the concentration of the target biomarker, the proposed platform can be applied to the quantification of the target biomarker in complex biological samples. The effectiveness of the proposed platform was tested on the detection of carcinoembryonic antigen (CEA) in serum. Experimental results revealed that the proposed platform could achieve quite reliable quantitative results for CEA in human serum samples with accuracy comparable to a commercial CEA ELISA Kit. Its limit of detection and limit of quantification for CEA were estimated to be 0.6 × 10-3 and 1.8 × 10-3 ng/mL, respectively, considerably lower than the corresponding values reported in literature. Due to its features of simplicity in design, extremely low background signal, high sensitivity and selectivity, the proposed method can be further developed to be a competitive alternative for the quantification of CEA and other protein biomarkers as well.


Assuntos
Nanopartículas , Biomarcadores , Humanos , Magnetismo , Peptídeos , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz
3.
R Soc Open Sci ; 7(7): 191653, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32874599

RESUMO

Density functional theory was employed to investigate the (111), (200), (210), (211) and (220) surfaces of CoS2. The surface energies were calculated with a sulfur environment using first-principle-based thermodynamics. It is founded that surfaces with metal atoms at their outermost layer have higher energy. The stoichiometric (220) surface terminated by two layer of sulfur atoms is most stable under the sulfur-rich condition, while the non-stoichiometric (211) surface terminated by a layer of Co atoms has the lower energy under the sulfur-poor environment. The electric structure results show that the front valence electrons of (200) surface are active, indicating that there may be some active sites on this face. There is an energy gap between the stoichiometric (220) and (211), which has low Fermi energy, indicating that their electronic structures are dynamically stable. Spin-polarized bands are calculated on the stoichiometric surfaces, and these two (200) and (210) surfaces are predicted to be noticeably spin-polarized. The Bravais-Friedel-Donnay-Harker (BFDH) method is adopted to predict crystal growth habit. The results show that the most important crystal planes for the CoS2 crystal growth are (111) and (200) planes, and the macroscopic morphology of CoS2 crystal may be spherical, cubic, octahedral, prismatic or plate-shaped, which have been verified by experiments.

4.
Technol Cancer Res Treat ; 19: 1533033820917959, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32281513

RESUMO

The objective of this article is to study the effect of inhibiting phosphatase and tensin homolog deleted chromatosome 10 gene on phosphoinositide 3-kinase/protein kinase B (Akt)/Forkhead homeobox O3a signaling pathway in human nasopharyngeal carcinoma HK-1 cells. Nasopharyngeal carcinoma HK-1 cell lines were divided into PTEN gene interference group (siPTEN), nonspecific small interfering RNA group (siNC), empty vector group (Vector), and no transfection control group (Normal). The mRNA and protein expression levels of PTEN, PI3K, p-Akt, and FoxO3a were detected by real-time fluorescence quantitative polymerase chain reaction and Western blot. Immunofluorescence was used to detect the subcellular localization of PTEN, PI3K, p-Akt, and FoxO3a in HK-1 cells. The proliferation of HK-1 cells was detected by MTT assay, and the apoptosis of HK-1 cells was detected by flow cytometry. Compared with the siNC group, the expression levels of PTEN, FoxO3a messenger RNA, and protein in the siPTEN group were significantly decreased (P < .05), while the expression levels of PI3K, p-Akt messenger RNA, and protein were significantly increased (P < .05). The growth rate of HK-1 cells in the siPTEN group was significantly higher than the siNC group (P < .05), while the apoptosis rate was significantly lower than that of the siNC group (P < .05). Small interfering RNA can inhibit the expression of PTEN in HK-1 cells, and PTEN can participate in the development of NPC by affecting PI3K/Akt/FoxO3a signaling pathway.


Assuntos
Classe I de Fosfatidilinositol 3-Quinases/metabolismo , Proteína Forkhead Box O3/metabolismo , Neoplasias Nasofaríngeas/patologia , PTEN Fosfo-Hidrolase/genética , Proteínas Proto-Oncogênicas c-akt/metabolismo , RNA Interferente Pequeno/genética , Apoptose/fisiologia , Linhagem Celular Tumoral , Movimento Celular/fisiologia , Proliferação de Células/fisiologia , Classe I de Fosfatidilinositol 3-Quinases/genética , Proteína Forkhead Box O3/genética , Humanos , Carcinoma Nasofaríngeo/genética , Carcinoma Nasofaríngeo/metabolismo , Carcinoma Nasofaríngeo/patologia , Neoplasias Nasofaríngeas/genética , Neoplasias Nasofaríngeas/metabolismo , PTEN Fosfo-Hidrolase/antagonistas & inibidores , PTEN Fosfo-Hidrolase/biossíntese , PTEN Fosfo-Hidrolase/metabolismo , Proteínas Proto-Oncogênicas c-akt/genética , RNA Interferente Pequeno/administração & dosagem , RNA Interferente Pequeno/metabolismo , Transdução de Sinais
5.
Diagn Cytopathol ; 47(11): 1197-1202, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31318494

RESUMO

Intrathyroidal thymic carcinoma is a malignant epithelial tumor of the thyroid gland with thymic epithelial differentiation; it is the malignant counterpart of ectopic thymoma of the thyroid gland and histologically resembles thymic carcinoma. Given its favorable prognostic course, preoperative cytological diagnosis is important to distinguish this neoplasm from other more aggressive thyroid neoplasms that can have similar or overlapping cytomorphological features. However, there is little information about the cytology of this disease available in the literature, even in the latest fourth edition of World Health Organization classification of tumors of endocrine organs. Here, we report a case of intrathyroidal thymic carcinoma exhibiting neuroendocrine differentiation in a 33-year-old woman and present a review of the literature.


Assuntos
Diferenciação Celular , Células Neuroendócrinas , Timoma , Neoplasias do Timo , Glândula Tireoide , Neoplasias da Glândula Tireoide , Adulto , Feminino , Humanos , Imuno-Histoquímica , Células Neuroendócrinas/metabolismo , Células Neuroendócrinas/patologia , Timoma/diagnóstico , Timoma/metabolismo , Timoma/patologia , Neoplasias do Timo/diagnóstico , Neoplasias do Timo/metabolismo , Neoplasias do Timo/patologia , Glândula Tireoide/metabolismo , Glândula Tireoide/patologia , Neoplasias da Glândula Tireoide/diagnóstico , Neoplasias da Glândula Tireoide/metabolismo , Neoplasias da Glândula Tireoide/patologia
6.
J Mass Spectrom ; 54(3): 250-257, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30672064

RESUMO

A facile mass spectrometric kinetic method for quantitative analysis of chiral compounds was developed by integrating mass spectrometry based on chemical derivatization and the spectral shape deformation quantitative theory. Chemical derivatization was employed to introduce diastereomeric environments to the chiral compounds of interest, resulting in different abundance distribution patterns of fragment ions of the derivatization products of enantiomers in mass spectrometry. The quantitative information of the chiral compounds of interest was extracted from complex mass spectral data by an advanced calibration model derived based on the spectral shape deformation quantitative theory. The performance of the proposed method was tested on the quantitative analysis of R-propranolol in propranolol tablets. Experimental results demonstrated that it could achieve accurate and precise concentration ratio predictions for R-propranolol with an average relative predictive error (ARPE) of about 4%, considerably better than the corresponding results of the mass spectrometric method based on chemical derivatization and the univariate ratiometric model (ARPE: about 12%). The limit of detection (LOD) and limit of quantification (LOQ) of the proposed method for the concentration ratio of R-propranolol were estimated to be 1.5% and 6.0%, respectively. The proposed method is complementary to the existing methods designed for the quantification of enantiomers such as the Cooks kinetic method.

7.
Curr Stem Cell Res Ther ; 13(8): 658-664, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30095059

RESUMO

There is increasing evidence reporting that as a common phenomenon in MetS relative diseases, insulin resistance (IR) is regarded as an independent etiological factor and a warning indicator of MetS occurrence. Therefore, for the special group (overweight or obesity), clinical regular monitoring of IR is an important basis for the prevention and early intervention of MetS relative diseases. This surveys reveals that human umbilical cord mesenchymal stem cells (HUC-MSCs)possess a kind of potential: it may become a possible theraphy for IR in type 2 diabetes mellitus (T2DM) and related diseases. Specific emphasis is focused on evaluating the improvement IR function of HUC-MSCs under the background of development in vitro and in vivo. Next, the action mechanisms of HUC-MSCs is discussed, and some of their advantages and disadvantages in the course of clinic application are presented. The final section highlights the application of HUC-MSCs in T2DM and relative diseases at this stage. Up to now, although many questions remain unresolved, we still consider that HUC-MSCs is one of the best therapy ameliorating IR in the future.


Assuntos
Resistência à Insulina , Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais/citologia , Cordão Umbilical/citologia , Diabetes Mellitus/terapia , Humanos
8.
Huan Jing Ke Xue ; 38(5): 1783-1791, 2017 May 08.
Artigo em Chinês | MEDLINE | ID: mdl-29965081

RESUMO

In Beijing, the chemical composition and component concentrations of volatile organic compounds (VOCs) were investigated during the municipal solid waste composting process using a portable gas chromatograph coupled with a mass spectrometer. The contributions of VOCs to the ozone formation potential were computed using the maximum incremental reactivity (MIR) scale and the propylene-equivalent concentration scale. The results showed that the concentrations of waste discharge in the sorting room, the first fermentation workshop, the second fermentation workshop, the compost product workshop, and the plant boundary were 10302.1, 15484.1, 929.9, 4693.6 and 370.4 µg·m-3, respectively. The main VOCs of the municipal solid waste composting plant were ethanol, limonene, and acetone. The propylene-equivalent concentrations of waste discharge in the sorting room, the first fermentation workshop, the second fermentation workshop, the compost product workshop, and the plant boundary were 25875.7, 4087.9, 378.0, 747.7 and 296.8 µg·m-3, whereas the O3 formation potentials computed using the MIR scale were 26979.3, 21168.3, 1469.3, 6439.6 and 455.8 µg·m-3. Reducing pollution by controlling the VOCs emission of waste discharge in the sorting room and the first fermentation workshop is important and can decrease the ozone formation potential. Given the accuracy and accessibility of the method, the MIR scale is more suitable for calculating the ozone formation potential of VOCs emitted from the municipal solid waste composting plant.

9.
J Air Waste Manag Assoc ; 66(9): 930-40, 2016 09.
Artigo em Inglês | MEDLINE | ID: mdl-27192607

RESUMO

UNLABELLED: On the basis of total temperature increase, normal dehydration, and maturity, the odor compositions of surface and internal piles in a well-run sewage sludge compost plant were analyzed using gas chromatography-mass spectrometry with a liquid nitrogen cooling system and a portable odor detector. Approximately 80 types of substances were detected, including 2 volatile inorganic compounds, 4 sulfur organic compounds, 16 benzenes, 27 alkanes, 15 alkenes, and 19 halogenated compounds. Most pollutants were mainly produced in the mesophilic and pre-thermophilic periods. The sulfur volatile organic compounds contributed significantly to odor and should be controlled primarily. Treatment strategies should be based on the properties of sulfur organic compounds. Hydrogen sulfide, methyl mercaptan, dimethyl disulfide, dimethyl sulfide, ammonia, and carbon disulfide were selected as core indicators. Ammonia, hydrogen sulfide, carbon disulfide, dimethyl disulfide, methyl mercaptan, dimethylbenzene, phenylpropane, and isopentane were designated as concentration indicators. Benzene, m-xylene, p-xylene, dimethylbenzene, dichloromethane, toluene, chlorobenzene, trichloromethane, carbon tetrachloride, and ethylbenzene were selected as health indicators. According to the principle of odor pollution indicator selection, dimethyl disulfide was selected as an odor pollution indicator of sewage sludge composting. Monitoring dimethyl disulfide provides a highly scientific method for modeling and evaluating odor pollution from sewage sludge composting facilities. IMPLICATIONS: Composting is one of the most important methods for sewage sludge treatment and improving the low organic matter content of many agricultural soils. However, odors are inevitably produced during the composting process. Understanding the production and emission patterns of odors is important for odor control and treatment. Core indicators, concentration indicators, and health indicators provide an index system to odor evaluation. An odor pollution indicator provides theoretical support for further modelling and evaluating odor pollution from sewage sludge composting facilities.


Assuntos
Poluentes Atmosféricos/análise , Poluição do Ar/análise , Monitoramento Ambiental/métodos , Odorantes/análise , Esgotos/química , Solo/química , Cromatografia Gasosa-Espectrometria de Massas
10.
J BUON ; 21(6): 1471-1475, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-28039710

RESUMO

PURPOSE: Borrmann type IV gastric cancer is still detected at an advanced stage and survival rates for these patients remain poor. The purpose of this study was to provide valuable prognostic and surgical information on patients with Borrmann type IV gastric cancer. METHODS: We compared the clinicopathological features of Borrmann type IV gastric cancer with those of other types of gastric cancer. Clinicopathologic features of patients with Borrmann type IV gastric cancer were evaluated as prognostic predictors by univariate and multivariate analyses. RESULTS: The results showed that Borrmann type IV gastric cancer had more advanced and unfavorable clinicopathological factors compared with other Borrman types. The 5-year overall survival rate was 16.7%, and the median survival 25 months. The 5-year overall survival rate was influenced by tumor size, depth of invasion, lymph node metastasis, and chemotherapy. Of these, independent prognostic factors were lymph node metastasis (present vs absent, risk ratio 1.855, p=0.002) and radical (R0) resection (no vs yes, risk ratio 2.035, p<0.001). A significant survival benefit for radical resection was observed, with a 5-year overall survival rate of 30.7% compared with non-radically resected cases (4.8%). CONCLUSIONS: Early detection and radical resection were essential to improve the prognosis of patients with Borrmann type IV gastric cancer.


Assuntos
Neoplasias Gástricas/patologia , Adulto , Idoso , Antineoplásicos/uso terapêutico , Distribuição de Qui-Quadrado , Feminino , Gastrectomia , Humanos , Estimativa de Kaplan-Meier , Metástase Linfática , Masculino , Pessoa de Meia-Idade , Análise Multivariada , Invasividade Neoplásica , Estadiamento de Neoplasias , Modelos de Riscos Proporcionais , Estudos Retrospectivos , Fatores de Risco , Neoplasias Gástricas/classificação , Neoplasias Gástricas/mortalidade , Neoplasias Gástricas/terapia , Fatores de Tempo , Resultado do Tratamento , Carga Tumoral
11.
Arch Virol ; 161(2): 405-16, 2016 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-26597185

RESUMO

The circulation of duck hepatitis A virus types 1 (DHAV-1) and 3 (DHAV-3) in Southeast Asia has resulted in a continuously changing epidemiological scenario. In this study, a duplex real-time PCR assay for simultaneous quantitative detection of DHAV-1 and DHAV-3 was established, and 200 liver samples from dead ducklings collected from 31 different flocks in Shandong province, China, were tested. Fifty-eight (29.0 %) samples from 13 flocks were positive for DHAV-1 single infection, 113 (56.5 %) samples from 13 other flocks were positive for DHAV-3 single infection, and 24 samples (12.0 %) from four flocks were positive for both viruses. DHAV-1 and DHAV-3 were detected with high viral loads in all of the organs tested (liver, spleen, pancreas, kidney, heart, thymus, bursa of Fabricius and brain). No significant difference in DHAV-1 and DHAV-3 viral loads was found between singly infected and coinfected samples, and there was no correlation between the viral loads of the two viruses and the age of dead ducklings. To the best of our knowledge, this is the first report about the in vivo distribution of DHAV-1 and DHAV-3 in clinically infected ducklings.


Assuntos
Patos/virologia , Vírus da Hepatite do Pato/isolamento & purificação , Hepatite Viral Animal/virologia , Infecções por Picornaviridae/veterinária , Estruturas Animais/virologia , Animais , Animais Recém-Nascidos , China/epidemiologia , Coinfecção/epidemiologia , Coinfecção/veterinária , Coinfecção/virologia , Hepatite Viral Animal/epidemiologia , Epidemiologia Molecular , Reação em Cadeia da Polimerase Multiplex , Infecções por Picornaviridae/epidemiologia , Infecções por Picornaviridae/virologia , Prevalência , Reação em Cadeia da Polimerase em Tempo Real , Carga Viral
12.
Genet Test Mol Biomarkers ; 18(8): 546-51, 2014 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-25014214

RESUMO

Spinal muscular atrophy (SMA) is a common autosomal recessive neuromuscular disorder caused by mutations of the survival of motor neuron 1 (SMN1) gene. Approximately 90-95% of SMA patients have a homozygous deletion of SMN1, and 5-10% of patients are believed to have subtle mutations. The molecular diagnosis of SMN1 subtle mutations is hampered by a highly homologous SMN2 gene. It is important to establish a rational molecular diagnostic procedure for SMN1 subtle mutations. We analyzed the SMN1 mutations in nine nonhomozygous patients by the following procedures: multiplex ligation-dependent probe amplification, genomic sequencing, T-A cloning on cDNA or genomic level, and/or real-time quantitative analysis. By the above molecular diagnostic procedure, six SMN1 subtle mutations, including c.5C>G(p.Ala2Gly), c.22_23 insA (p.Ser8LysfsX23), c.40G>T(p.Glu14X), c.43C>T(p.Gln15X), c.683T>A(p.Leu228X), and c.56delT(p.Val19GlyfsX21), were identified in nine Chinese patients. p.Glu14X has not been reported previously. Compared with the level of full-length SMN1 transcripts in the healthy carriers (14.1±4.5), the patient with p.Ala2Gly had no significant reduction (13.9±3.64, p=0.955). However, the levels in the patients carrying other mutations were significantly reduced (0.27±0.139 to 13.9±3.64, p=0.000-0.004). We present a reliable and rational diagnostic procedure for SMN1 subtle mutations, which would be helpful in molecular diagnosis of SMA compound heterozygotes. Our work extends the SMN1 mutation spectrum.


Assuntos
Transtornos Heredodegenerativos do Sistema Nervoso/genética , Atrofia Muscular Espinal/genética , Mutação de Sentido Incorreto , Patologia Molecular/métodos , Proteína 1 de Sobrevivência do Neurônio Motor/genética , Substituição de Aminoácidos , China , Feminino , Transtornos Heredodegenerativos do Sistema Nervoso/diagnóstico , Humanos , Masculino , Atrofia Muscular Espinal/diagnóstico , Proteína 2 de Sobrevivência do Neurônio Motor/genética
13.
Dis Markers ; 2014: 912182, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24591770

RESUMO

BACKGROUND: The molecular work-up of thyroid nodules from fine needle aspiration samples has given clinicians a new level of diagnostic information. The aim of the present study was to evaluate the utility of molecular analysis in thyroid fine needle aspiration samples from a Chinese population. METHODS: Specimens were collected from thyroid nodules by fine needle aspiration. Cytology diagnosis and genes analysis were performed and correlated with histology outcome. RESULTS: A total of 83 patients with thyroid nodules were enrolled, including 20 benign lesions and 63 papillary carcinomas. BRAF and RAS mutations and RET/PTC gene rearrangements were found in 65.1%, 0%, and 1.6% of papillary carcinomas, respectively. No gene alterations were found in benign lesions. The combination of BRAF testing and cytology improved the accuracy of cytology from 69.9% to 89.2% (P < 0.05). Moreover, BRAF testing confirmed 82.4% of papillary carcinomas with suspicious cytology and identified 33.3% of papillary carcinomas with atypia cytology. CONCLUSIONS: Of the three candidate markers, BRAF testing showed diagnostic utility in fine needle aspiration. Combining BRAF testing with cytology improves the accuracy of fine needle biopsy. Those who have positive BRAF and malignant or suspicious malignant cytology can undergo thyroidectomy without a frozen section.


Assuntos
Carcinoma/diagnóstico , Glândula Tireoide/patologia , Neoplasias da Glândula Tireoide/diagnóstico , Nódulo da Glândula Tireoide/diagnóstico , Biópsia por Agulha Fina , Carcinoma/genética , Carcinoma Papilar , China , Análise Mutacional de DNA , Humanos , Técnicas de Diagnóstico Molecular , Proteínas Proto-Oncogênicas B-raf/genética , Câncer Papilífero da Tireoide , Neoplasias da Glândula Tireoide/genética , Nódulo da Glândula Tireoide/genética , Proteínas ras/genética
14.
J Virol Methods ; 192(1-2): 12-7, 2013 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-23624117

RESUMO

Infection with duck hepatitis A virus (DHAV) causes an acute, rapidly spreading, and fatal disease of young ducklings. DHAV type 1 (DHAV-1) and type 3 (DHAV-3) have been identified in China. In this study, a duplex RT-PCR assay was developed to identify DHAV-1 and DHAV-3 with mixed infection. The method was shown to be high specificity and sensitivity. The minimum detection limit of the method has been determined to be 10pg total RNA templates extracted from duck liver samples or 10² copies viral RNA of DHAV-1 and DHAV-3 respectively. Using the method, from 60 clinical liver samples of 26 duckling flocks in Shandong, Guangdong, Sichuan and Henan provinces of China, 15 (57.7%) flocks were identified as mixed infection of DHAV-1 and DHAV-3, and 9 (34.6%) flocks were DHAV-1 or DHAV-3 single infection. Among them, 38.3% (23/60) of duckling samples were detected as mixed infection of DHAV-1 and DHAV-3, and 48.3% (29/60) of samples were DHAV-1 or DHAV-3 single infection. These results indicated that the improved duplex RT-PCR method provides a rapid and cost-effective laboratory differential diagnosis for mixed infection of DHAV-1 and DHAV-3 in ducklings.


Assuntos
Vírus da Hepatite do Pato/classificação , Vírus da Hepatite do Pato/isolamento & purificação , Reação em Cadeia da Polimerase Multiplex/métodos , Infecções por Picornaviridae/veterinária , Doenças das Aves Domésticas/diagnóstico , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Medicina Veterinária/métodos , Animais , China , Coinfecção/diagnóstico , Coinfecção/veterinária , Coinfecção/virologia , Patos , Vírus da Hepatite do Pato/genética , Infecções por Picornaviridae/diagnóstico , Infecções por Picornaviridae/virologia , Doenças das Aves Domésticas/virologia , Sensibilidade e Especificidade , Virologia/métodos
15.
Bing Du Xue Bao ; 29(5): 522-8, 2013 Sep.
Artigo em Chinês | MEDLINE | ID: mdl-24386841

RESUMO

To reveal the genetic variation of the viral protein 1 (VP1) gene of the duck hepatitis A virus type 3 (DHAV-3), the VP1 gene of 13 virulent DHAV-3 strains isolated from Shandong province of China in 2012 were amplified by RT-PCR, sequenced and analyzed. The results showed that all the VP1 genes of the 13 isolates contained 720 nucleotides encoding 240 amino acids, and shared with nucleotide identities of 94. 6%-99.9% and amino acid identities of 95.0%-100%. The nucleotide and amino acid sequence homologies between the 13 DHAV-3 isolates and other 31 DHAV-3 reference strains were 92.5%-100% and 90. 8%-100%, respectively. Phylogenetic analysis showed that the VP1 gene of DHAV-3 had distinct geographical characteristics. Distribution of genotypes of the 44 DHAV-3 strains was as follows: except the vaccine strain B63, all the other Chinese isolates belonged to genotype I (GI), Vietnamese wild isolates mainly belonged to subtype 1 (S1) of genotype II (GII), and all Korean isolates belonged to subtype 2 (S2) of GII.


Assuntos
Proteínas do Capsídeo/genética , Vírus da Hepatite do Pato/genética , Vírus da Hepatite do Pato/isolamento & purificação , Hepatite Viral Animal/virologia , Infecções por Picornaviridae/veterinária , Doenças das Aves Domésticas/virologia , Sequência de Aminoácidos , Animais , Proteínas do Capsídeo/química , China , Patos , Vírus da Hepatite do Pato/classificação , Dados de Sequência Molecular , Filogenia , Infecções por Picornaviridae/virologia
16.
Virology ; 436(1): 112-7, 2013 Feb 05.
Artigo em Inglês | MEDLINE | ID: mdl-23174505

RESUMO

The capsid protein (CP) of duck circovirus (DuCV) is the major immunogenic protein and has a high proportion of arginine residues concentrated at the N terminus of the protein, which inhibits efficient mRNA translation in prokaryotic expression systems. In this study, we investigated the subcellular distribution of DuCV CP expressed via recombinant baculoviruses in Sf9 cells and the DNA binding activities of the truncated recombinant DuCV CPs. The results showed that two independent bipartite nuclear localization signals (NLSs) situated at N-terminal 1-17 and 18-36 amino acid residue of the CP. Moreover, two expression level regulatory signals (ELRSs) and two DNA binding signals (DBSs) were also mapped to the N terminus of the protein and overlapped with the two NLSs. The ability of CP to bind DNA, coupled with the karyophilic nature of this protein, strongly suggests that it may be responsible for nuclear targeting of the viral genome.


Assuntos
Proteínas do Capsídeo/química , Proteínas do Capsídeo/metabolismo , Circovirus/metabolismo , Sinais de Localização Nuclear/metabolismo , Animais , Baculoviridae/genética , Proteínas do Capsídeo/genética , Linhagem Celular , Núcleo Celular/metabolismo , Circovirus/genética , Proteínas de Ligação a DNA/química , Proteínas de Ligação a DNA/metabolismo , Patos/virologia , Genoma Viral , Sinais de Localização Nuclear/química , Sinais de Localização Nuclear/genética , Proteínas Recombinantes/metabolismo , Células Sf9 , Transdução de Sinais , Replicação Viral
17.
Mol Biol Rep ; 40(3): 2473-83, 2013 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-23187740

RESUMO

Substantial evidence has demonstrated that platelet-derived growth factor-D (PDGF-D) is tightly associated with the development and progression of tumors. However, its biological functions in esophageal squamous cell carcinoma (ESCC) remain to be delineated. In this study, we found that expressions of PDGF-D mRNA and protein in ESCC tissues and cells were significantly higher than that in normal esophageal epithelial tissues (P < 0.05), further investigation showed that PDGF-D protein level in EC1 cells was obviously higher than those in EC9706 and Eca109 cells (P < 0.05). Elevated PDGF-D level was closely associated with TNM staging, tumor differentiation and lymph node metastasis (P < 0.05), but not related to the patients' age and gender (P > 0.05). In addition, down-regulation of PDGF-D expression markedly inhibited proliferation, reduced invasion and induced apoptosis in EC1 cells. More importantly, reduced PDGF-D level evoked the down-regulation of p65 and p-IκBα proteins and elevation of IκBα protein of NF-κB pathway, accompanied with the decreases of bcl-2 and MMP-9 protein expressions and increases of bax protein level and caspase-3 activities. Correctively, our data suggest that PDGF-D plays pivotal roles in the development and progression of ESCC, and combinations with PDGF-D and NF-κB pathway may be effective and feasible molecular targets for therapy of ESCC.


Assuntos
Apoptose/genética , Carcinoma de Células Escamosas/genética , Carcinoma de Células Escamosas/metabolismo , Neoplasias Esofágicas/genética , Neoplasias Esofágicas/metabolismo , Regulação Neoplásica da Expressão Gênica , Linfocinas/genética , NF-kappa B/metabolismo , Fator de Crescimento Derivado de Plaquetas/genética , Transdução de Sinais , Adulto , Idoso , Carcinoma de Células Escamosas/patologia , Caspase 3/metabolismo , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células , Neoplasias Esofágicas/patologia , Carcinoma de Células Escamosas do Esôfago , Esôfago , Feminino , Humanos , Linfocinas/metabolismo , Masculino , Pessoa de Meia-Idade , Membrana Mucosa/metabolismo , Membrana Mucosa/patologia , Invasividade Neoplásica , Estadiamento de Neoplasias , Fator de Crescimento Derivado de Plaquetas/metabolismo , Interferência de RNA , RNA Mensageiro/genética , RNA Mensageiro/metabolismo
18.
Vet Microbiol ; 161(3-4): 331-3, 2013 Jan 25.
Artigo em Inglês | MEDLINE | ID: mdl-22906528

RESUMO

In 2009, an influenza virus (IV), A/canine/Shandong/JT01/2009 (CA/SD/JT01/09), was isolated from the dog exhibiting respiratory signs in China, and was a novel H5N2. Intraspecies transmission of the virus in dog population had thus far remained unclear. To determine whether the novel H5N2 was transmitted among dogs, we conducted contact exposure and inoculation experiments. Susceptible dogs were housed in the room which the novel H5N2 infected dogs were housed in. As a result, the direct contact resulted in intraspecies transmission. Most of the infected dogs and the sentinel animals developed mild respiratory syndrome, including transient increased body temperatures, conjunctivitis, sneezing, nasal discharge and mild coughing, virus shedding and seroconversion, but no fatal disease. These data suggest that dogs may play a role in transmission and spread of influenza virus.


Assuntos
Doenças do Cão/transmissão , Doenças do Cão/virologia , Vírus da Influenza A Subtipo H5N2/fisiologia , Infecções por Orthomyxoviridae/veterinária , Animais , China , Cães , Vírus da Influenza A Subtipo H5N2/isolamento & purificação , Infecções por Orthomyxoviridae/transmissão , Infecções por Orthomyxoviridae/virologia , Eliminação de Partículas Virais
19.
Vet Microbiol ; 159(1-2): 251-6, 2012 Sep 14.
Artigo em Inglês | MEDLINE | ID: mdl-22537707

RESUMO

Duck circovirus (DuCV) is classified in the genus Circovirus of the Circoviridae family. Two major open reading frames (ORFs), encoding the replicase (ORF1/rep) and the capsid protein (ORF2/cap), have been recognized for DuCV. Sequence analysis show that another major conserved ORF (named ORF3) is located in the complementary strand of ORF1/rep of DuCV, and its function remains to be investigated. In this study, the ORF3 of DuCV was expressed in recombinant baculovirus-infected Sf9 cells. By IFA and Western blot analysis, the ORF3 protein was positive for the sera from ducks infected with DuCV. The percentages of apoptotic cells of the Sf9 cells infected with the recombinant baculovirus encoding ORF3 of DuCV were significantly higher than (P<0.05) that of the Sf9 cells infected with wild-type baculovirus at 24, 48 and 72 h postinfection. Based on our knowledge, we deduced that the ORF3 protein of DuCV might play an important role in viral pathogenesis via its apoptotic activity.


Assuntos
Apoptose/genética , Circovirus/genética , Circovirus/metabolismo , Proteínas Virais/genética , Animais , Baculoviridae/genética , Baculoviridae/crescimento & desenvolvimento , Western Blotting , Linhagem Celular , Circovirus/classificação , Circovirus/patogenicidade , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Células Sf9
20.
Vet Microbiol ; 155(2-4): 409-16, 2012 Mar 23.
Artigo em Inglês | MEDLINE | ID: mdl-22033043

RESUMO

An influenza virus, A/canine/Shandong/JT01/2009, has been isolated from a dog exhibiting classical flu signs in China. HAI and NAI assays subtyped A/canine/Shandong/JT01/2009 as a H5N2 like virus. Phylogenetic reconstructions indicated strong relationships with viruses from various hosts and dispersed geographic locations. These analyses indicate A/canine/Shandong/JT01/2009 is a novel virus generated by complex reassortment of the viral segments.


Assuntos
Doenças do Cão/virologia , Vírus da Influenza A Subtipo H5N2/classificação , Vírus da Influenza A Subtipo H5N2/genética , Infecções por Orthomyxoviridae/veterinária , Animais , China , Cães , Vírus da Influenza A Subtipo H5N2/isolamento & purificação , Infecções por Orthomyxoviridae/virologia , Filogenia
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