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1.
Vet Res ; 55(1): 82, 2024 Jun 27.
Artigo em Inglês | MEDLINE | ID: mdl-38937820

RESUMO

Respiratory diseases constitute a major health problem for ruminants, resulting in considerable economic losses throughout the world. Parainfluenza type 3 virus (PIV3) is one of the most important respiratory pathogens of ruminants. The pathogenicity and phylogenetic analyses of PIV3 virus have been reported in sheep and goats. However, there are no recent studies of the vaccination of sheep or goats against PIV3. Here, we developed a purified inactivated ovine parainfluenza virus type 3 (OPIV3) vaccine candidate. In addition, we immunized sheep with the inactivated OPIV3 vaccine and evaluated the immune response and pathological outcomes associated with OPIV3 TX01 infection. The vaccinated sheep demonstrated no obvious symptoms of respiratory tract infection, and there were no gross lesions or pathological changes in the lungs. The average body weight gain significantly differed between the vaccinated group and the control group (P < 0.01). The serum neutralization antibody levels rapidly increased in sheep post-vaccination and post-challenge with OPIV3. Furthermore, viral shedding in nasal swabs and viral loads in the lungs were reduced. The results of this study suggest that vaccination with this candidate vaccine induces the production of neutralizing antibodies and provides significant protection against OPIV3 infection. These results may be helpful for further studies on prevention and control strategies for OPIV3 infections.


Assuntos
Infecções por Respirovirus , Doenças dos Ovinos , Vacinas de Produtos Inativados , Vacinas Virais , Animais , Ovinos , Infecções por Respirovirus/veterinária , Infecções por Respirovirus/prevenção & controle , Infecções por Respirovirus/virologia , Infecções por Respirovirus/imunologia , Vacinas de Produtos Inativados/imunologia , Doenças dos Ovinos/prevenção & controle , Doenças dos Ovinos/virologia , Doenças dos Ovinos/imunologia , Vacinas Virais/imunologia , Respirovirus/imunologia , Imunogenicidade da Vacina , Vacinação/veterinária
2.
BMC Vet Res ; 20(1): 209, 2024 May 18.
Artigo em Inglês | MEDLINE | ID: mdl-38760785

RESUMO

BACKGROUND: Bovine coronavirus (BCoV) is implicated in severe diarrhea in calves and contributes to the bovine respiratory disease complex; it shares a close relationship with human coronavirus. Similar to other coronaviruses, remarkable variability was found in the genome and biology of the BCoV. In 2022, samples of feces were collected from a cattle farm. A virus was isolated from 7-day-old newborn calves. In this study, we present the genetic characteristics of a new BCoV isolate. The complete genomic, spike protein, and nucleocapsid protein gene sequences of the BCoV strain, along with those of other coronaviruses, were obtained from the GenBank database. Genetic analysis was conducted using MEGA7.0 and the Neighbor-Joining (NJ) method. The reference strains' related genes were retrieved from GenBank for comparison and analysis using DNAMAN. RESULTS: The phylogenetic tree and whole genome consistency analysis showed that it belonged to the GIIb subgroup, which is epidemic in Asia and America, and was quite similar to the Chinese strains in the same cluster. Significantly, the S gene was highly consistent with QH1 (MH810151.1) isolated from yak. This suggests that the strain may have originated from interspecies transmission involving mutations of wild strains. The N gene was conserved and showed high sequence identity with the epidemic strains in China and the USA. CONCLUSIONS: Genetic characterization suggests that the isolated strain could be a new mutant from a wild-type lineage, which is in the same cluster as most Chinese epidemic strains but on a new branch.


Assuntos
Doenças dos Bovinos , Infecções por Coronavirus , Coronavirus Bovino , Genoma Viral , Filogenia , Animais , Bovinos , Coronavirus Bovino/genética , Coronavirus Bovino/isolamento & purificação , China/epidemiologia , Doenças dos Bovinos/virologia , Doenças dos Bovinos/epidemiologia , Infecções por Coronavirus/veterinária , Infecções por Coronavirus/virologia , Infecções por Coronavirus/epidemiologia , Fezes/virologia , Glicoproteína da Espícula de Coronavírus/genética , Animais Recém-Nascidos
3.
Microb Pathog ; 181: 106155, 2023 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-37301331

RESUMO

Type I interferon has great broad-spectrum antiviral ability and immunomodulatory function, and its receptors are expressed in almost all types of cells. Bovine viral diarrhea virus (BVDV) is an important pathogen causing significant economic losses in cattle. In this study, a recombinant expression plasmid carrying bovine interferon-α(BoIFN-α)gene was constructed and transformed into E. coli BL21 (DE3) competent cells. SDS-PAGE and Westernblotting analysis showed that the recombinant BoIFN-α protein (rBoIFN-α) was successfully expressed. It is about 36KD and exists in the form of inclusion body. When denatured, purified and renatured rBoIFN-α protein stimulated MDBK cells, the expression of interferon stimulating genes (ISGs) such as ISG15, OAS1, IFIT1, Mx1 and IFITM1 were significantly up-regulated, and reached the peak at 12 h (P< 0.001). MDBK cells were infected with BVDV with moi of 0.1 and 1.0, respectively. The virus proliferation was observed after pretreatment with rBoIFN-α protein and post-infection treatment. The results showed that the denatured, purified and renatured BoIFN-α protein had good biological activity and could inhibit the replication of BVDV in MDBK cells in vitro, which provided a basis for BoIFN-α as an antiviral drug, immune enhancer and clinical application of BVDV.


Assuntos
Vírus da Diarreia Viral Bovina , Interferon Tipo I , Animais , Bovinos , Escherichia coli , Interferon-alfa/genética , Interferon-alfa/farmacologia , Interferon-alfa/metabolismo , Antivirais/uso terapêutico , Interferon Tipo I/metabolismo , Vírus da Diarreia Viral Bovina/genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/farmacologia , Proteínas Recombinantes/metabolismo
4.
Virol J ; 19(1): 33, 2022 02 23.
Artigo em Inglês | MEDLINE | ID: mdl-35197095

RESUMO

BACKGROUND: Rubella virus (RV) is the causative agent of rubella or German measles. Although most infections cause only mild self-limited measles-like illness, the infection in pregnant women can cause severe foetal malformation or even miscarriage, especially in the first 3 months of pregnancy. Therefore, it is of great practical significance to establish a simple and sensitive RV detection method. METHODS: The partial epitopes of the E1 and E2 proteins from Rubella Virus were selected as the target sites, the sequence of the selected antigenic sites of the E1 and E2 were linked by a linker. The expression plasmid P6T was constructed by inserting the gene into PET-32A + with a histidine Tag. The P6 protein was induced and expressed in Escherichia coli L21 (DE3) and purified by nickel column affinity. The protein P6 antigen was identified by Western blotting analysis, and an anti-P6 antibody ELISA was established to test known serum samples to evaluate the capability of this method. RESULTS: After purification, the concentration and purity of the protein P6 were 0.283 mg/mL and more than 80%, respectively. Western blotting analysis showed that the protein P6 could react with rubella virus positive serum. By ELISA, 36 negative sera and 58 positive sera were detected. The coincidence rate, specificity and sensitivity of the ELISA were 86.2%, 88.89% and 84.48%, respectively. The P6 ELISA with a kappa coefficient of 0.715, P < 0.05, indicated excellent consistency. CONCLUSIONS: The protein P6 with excellent antigenicity obtained from prokaryotic expression followed by chromatography purification could prove useful for early diagnosis of RV infection.


Assuntos
Vírus da Rubéola , Rubéola (Sarampo Alemão) , Anticorpos Antivirais , Western Blotting , Ensaio de Imunoadsorção Enzimática/métodos , Feminino , Humanos , Imunoglobulina G , Gravidez , Rubéola (Sarampo Alemão)/diagnóstico , Vírus da Rubéola/genética
5.
BMC Vet Res ; 18(1): 295, 2022 Jul 29.
Artigo em Inglês | MEDLINE | ID: mdl-35906644

RESUMO

BACKGROUND: The outbreak of Lumpy skin disease (LSD) in cattle caused by LSD virus (LSDV) was first reported in August 2019 in China. Since then, several LSD outbreaks have been reported in seven different provinces of China. Until now, several Lumpy skin disease virus (LSDV) strains from China have been reported and sequenced including LSDV/Xinjiang/2019 (MN598005.1), China/GD01/2020 (MW355944.1), and LSDV/Hongkong/2021 (MW732649.1). In October 2020, more than 1,700 cattle imported from Chile arrived in Xilingol, Inner Mongolia, and were diagnosed with LSD. Currently, limited data on the origin of the virus is available. METHODS: Nucleotide sequences of the ORF11, ORF36, ORF74, ORF117, ORF126 genes and the complete genome of LSDV strains and isolates were downloaded from NCBI database. MEGA7.0 was used to perform phylogenetic analysis with Neighbor-Joining (NJ). DNASTAR software is used to analyze homologous comparison analysis with related genes of reference strains included in Genbank. RESULTS: Compared with other strains isolated from China, the results of full genome sequence analysis showed the LSDV/NMG/2020 strain belonged to the recombinant strains. The LSDV/NMG/2020 strain is different from the current LSDV field isolates in Africa, the Middle East, Europe, and the newly emerged LSDV Russia variants. Based on the identities of P32, RPO30, EEV, GPCR and LSDV117 genes (99.8%, 99%, 99.8%, 99% and 98.7%), the sub-cluster recombinant containing LSDV/NMG/2020 strain is phylogenetically closer to the Russia strain (Saratov/2017). CONCLUSIONS: In this study, we reported a new isolated LSDV strain named LSDV/NMG/2020. The results of genomic characterization and phylogenetic analysis demonstrated that the LSDV/NMG/2020 isolate was a vaccine-like recombinant strain.


Assuntos
Doenças dos Bovinos , Doença Nodular Cutânea , Vírus da Doença Nodular Cutânea , Animais , Bovinos , Doenças dos Bovinos/epidemiologia , Surtos de Doenças/veterinária , Doença Nodular Cutânea/epidemiologia , Filogenia
6.
Sensors (Basel) ; 22(4)2022 Feb 14.
Artigo em Inglês | MEDLINE | ID: mdl-35214370

RESUMO

In the arid grasslands of northern China, unreasonable grazing methods can reduce the water content and species numbers of grassland vegetation. This project uses solar-powered GPS collars to obtain track data for sheep grazing. In order to eliminate the trajectory data of the rest area and the drinking area, the kernel density analysis method was used to cluster the trajectory point data. At the same time, the vegetation index of the experimental area, including elevation, slope and aspect data, was obtained through satellite remote sensing images. Therefore, using trajectory data and remote sensing image data to establish a neural network model of grazing intensity of sheep, the accuracy of the model could be high. The results showed that the best input parameters of the model were the combination of vegetation index, sheep weight, duration, moving distance and ambient temperature, where the coefficient of determination R2=0.97, and the mean square error MSE = 0.73. The error of grazing intensity obtained by the model is the smallest, and the spatial-temporal distribution of grazing intensity can reflect the actual situation of grazing intensity in different locations. Monitoring the grazing behavior of sheep in real time and obtaining the spatial-temporal distribution of their grazing intensity can provide a basis for scientific grazing.


Assuntos
Pradaria , Animais , China , Ovinos
7.
Microb Pathog ; 154: 104854, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33757898

RESUMO

Caprine parainfluenza virus type 3 (CPIV3) was first identified in goats named JS2013 in China. In 2019, a sheep herd broke a disease with respiratory disease in Hebei province, China. In order to confirm the pathogen of the disease, the nasal swabs, stool swabs and blood samples were collected from the sheep. Virus isolation was performed on MDBK cells and identification was conducted by RT-PCR. The complete genome of the isolate was sequenced and phylogenetic analyzed. In order to evaluate the pathogenicity of the virus, five seronegative sheep were experimental infected with the virus suspension. The phylogenetic analyses based on the complete genome and the M gene indicated that the isolate strain was distinguished distinct from previously reported CPIV3 lineage of JS2013. The virus-inoculated sheep displayed the syndrome with depression, cough, and fever. Virus shedding were detected by RT-PCR from nasal swabs. All infected showed virus shedding during 2 - 21dpi and viremia could be detected in serum samples. Gross pathological assessment of sheep in infected group showed gross lesion in the lungs. Histopathological observation results indicated that lungs had mild to moderate interstitial pneumonia, with thickened alveolar walls, decreased alveolar space, and increased amounts of inflammatory cells infiltration. This is the first report of pathogenicity of the novel lineage of sheep-derived CPIV3. The results would be helpful for further studies on the prevention and control strategies for CPIV3 infections in goat and sheep.


Assuntos
Doenças das Cabras , Infecções por Respirovirus , Doenças dos Ovinos , Animais , China , Cabras , Vírus da Parainfluenza 3 Humana/genética , Filogenia , Infecções por Respirovirus/veterinária , Ovinos , Virulência
8.
Biochem Biophys Res Commun ; 525(2): 326-333, 2020 04 30.
Artigo em Inglês | MEDLINE | ID: mdl-32093887

RESUMO

Spexin (SPX) acts as a neuropeptide with pleiotropic functions that can participate in anxiety regulation. Corticotropin releasing factor (CRF) is widely expressed in brain tissues and associated with depression and anxiety and addiction. With the anxious mice under chronic unpredictable stress, we found SPX mRNA expression level in the hippocampus of the brain was significantly reduced, while local CRF mRNA expression level was increased. Furthermore, CRF injection in the hippocampus could also decrease SPX mRNA expression levels in hippocampus and other brain tissues, including pituitary and hypothalamus. With the primary mouse hippocampal cell model, CRF treatment could decrease SPX mRNA expression at hippocampal cell level and this inhibitory effect was mediated only by corticotropin releasing factor receptor 2 (CRFR2) but not corticotropin releasing factor receptor 1 (CRFR1). In HEK293 cells with CRFR2 over-expression, CRF could also inhibit SPX promoter activity coupling with AC/cAMP/PKA and MEK1/2/Erk1/2 cascades. In addition, Epac was also involved with the CRF-repressed SPX promoter activity and cross-talked with MEK1/2/Erk1/2 pathway. CRF could inhibit SPX gene expression in mouse hippocampus via transcriptional activation at the promoter level with coupling of AC/cAMP and MEK1/2/Erk1/2 signaling, which will be relevant to the anxiety response mediated by SPX in central nervous system.


Assuntos
Ansiedade , Hormônio Liberador da Corticotropina/farmacologia , Hipocampo/metabolismo , Hormônios Peptídicos/fisiologia , Animais , Regulação da Expressão Gênica/efeitos dos fármacos , Células HEK293 , Humanos , Camundongos , Hormônios Peptídicos/efeitos dos fármacos , Hormônios Peptídicos/genética , Regiões Promotoras Genéticas/efeitos dos fármacos , RNA Mensageiro/efeitos dos fármacos , RNA Mensageiro/metabolismo , Receptores de Hormônio Liberador da Corticotropina , Transdução de Sinais
9.
Arch Biochem Biophys ; 688: 108366, 2020 07 30.
Artigo em Inglês | MEDLINE | ID: mdl-32387473

RESUMO

Preeclampsia (PE) is a pregnancy-related syndrome and has become the leading cause of maternal and neonatal morbidity and mortality. LncRNA has been elucidated to play critical roles in the phenotype of trophoblast cells. However, the effect of AK002210 has not been reported. We aim to investigate the effect of AK002210 on the phenotype of trophoblast cells. Quantitative reverse transcription PCR was used to assess the gene expression. CCK-8 assay was used to evaluate the cell proliferation. Transwell assay was performed to detect the migration and invasion of trophoblast cells. Luciferase assay and rescue experiment were carried out to verify the interaction between miR-590-3p and AK002210 as well as NLR family apoptosis inhibitory protein (NAIP). The results revealed that AK002210 promoted the proliferation, migration and invasion of trophoblast cell while AK002210 knockdown inhibited that. Mechanically, we found that AK002210 was targeted by miR-590-3p. Moreover, miR-590-3p also directly targets NAIP which served as a ceRNA of AK002210. Rescue experiment showed that miR-590-3p reversed the effect of AK002210 which further confirmed their interaction. Moreover, AK002210 was proved to participated in the regulation of ERK/MMP-2 signal axis. In conclusion, we found that AK002210 knockdown may play a critical role in the progression of PE via miR-590-3p/NAIP and ERK/MMP signaling. It has potential to be a novel prognostic or therapeutic marker of PE.


Assuntos
Movimento Celular/fisiologia , Proliferação de Células/fisiologia , Pré-Eclâmpsia/fisiopatologia , RNA Longo não Codificante/fisiologia , Transdução de Sinais/fisiologia , Trofoblastos/fisiologia , Apoptose/fisiologia , Linhagem Celular , Regulação para Baixo , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Feminino , Técnicas de Silenciamento de Genes , Humanos , Metaloproteinase 2 da Matriz/metabolismo , MicroRNAs/metabolismo , Proteína Inibidora de Apoptose Neuronal/metabolismo , Placenta/metabolismo , Pré-Eclâmpsia/metabolismo , Gravidez , RNA Longo não Codificante/genética , RNA Longo não Codificante/metabolismo
10.
Mikrochim Acta ; 187(2): 132, 2020 01 15.
Artigo em Inglês | MEDLINE | ID: mdl-31942660

RESUMO

The multifunctional hemin@carbon dot hybrid nanozymes (hemin@CD) with simultaneous peroxidase-like activity and fluorescence signalling property was prepared for the first time. Based on these properties, hemin@CD was applied to develop a dual-channel fluorescent probe for H2O2 and H2O2-based biocatalytic systems. By virtue of the peroxidase-like activity, hemin@CD can catalyze the oxidative coupling of 4-aminoantipyrine with phenol in the presence of H2O2 to form a pink-red quinoneimine dye with a maximum absorbance at 505 nm. Under the excitation wavelength of 480 nm, the green fluorescence of hemin@CD peaks at 540 nm and is quenched by the generated quinoneimine dye due to an inner filter effect, and also by H2O2 because of dynamic quenching. Thus, a colorimetric and fluorimetric dual-channel optical probe for H2O2 is obtained. Due to the glucose/xanthine transformations under formation of H2O2 by the relevant oxidase catalysis, the probe can be applied for detection of glucose and xanthine. The colorimetric detection limits for H2O2, glucose and xanthine are 0.11, 0.15, 0.11 µM, and the and fluorimetric detection limits are 0.15, 0.15, 0.12 µM, respectively. Graphical abstractSchematic representation of the colorimetric and fluorimetric dual probe for H2O2, glucose and xanthine based on the multifunctional emin@carbon dot) hybrid nanozymes with simultaneous peroxidase-like activity and fluorescence signalling property.


Assuntos
Glucose/análise , Peróxido de Hidrogênio/análise , Xantina/análise , Biocatálise , Carbono , Colorimetria/métodos , Colorimetria/normas , Corantes Fluorescentes/química , Fluorometria/métodos , Fluorometria/normas , Hemina , Limite de Detecção , Mimetismo Molecular , Peroxidase/metabolismo
11.
Analyst ; 144(9): 2994-3004, 2019 May 07.
Artigo em Inglês | MEDLINE | ID: mdl-30892312

RESUMO

The human telomerase reverse transcriptase catalytic subunit (hTERT) is the rate-limiting subunit of the telomerase holoenzyme. Down-regulating the expression of hTERT mRNA by antisense oligonucleotides would reduce the expression of hTERT, inhibit telomerase activity, and impair the growth of cancer cells in vitro. In this work, we propose a locked nucleic acid-functionalized gold nanoparticle flare probe (AuNP-probe). After transferring these probes into cells by endocytosis of the gold nanoparticles, the binding process of the antisense locked nucleic acid with hTERT mRNA along with gene regulation can be visualized by fluorescence recovery of flare-sequences. A significant decline in hTERT mRNA levels and the hTERT content occurred in cancer cells after treatment with the AuNP-probes, and only approximately 25% of the original level of hTERT mRNA remained after 72 h. AuNP-probe treated cancer cells were arrested in the G1 phase of the cell cycle and underwent apoptosis; cell viability decreased obviously compared with that of telomerase-negative normal cells.


Assuntos
DNA/química , Corantes Fluorescentes/química , Ouro/química , Nanopartículas Metálicas/química , RNA Mensageiro/metabolismo , Telomerase/metabolismo , Antineoplásicos/química , Antineoplásicos/farmacologia , Antineoplásicos/toxicidade , Apoptose/efeitos dos fármacos , Carbocianinas/química , Catequina/análogos & derivados , Catequina/farmacologia , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , DNA/toxicidade , Regulação para Baixo , Inibidores Enzimáticos/farmacologia , Fluorescência , Pontos de Checagem da Fase G1 do Ciclo Celular/efeitos dos fármacos , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Nanopartículas Metálicas/toxicidade , Microscopia Confocal/métodos , Microscopia de Fluorescência/métodos , Hibridização de Ácido Nucleico , Oligonucleotídeos/química , Oligonucleotídeos/genética , Oligonucleotídeos/toxicidade , Oligonucleotídeos Antissenso/química , Oligonucleotídeos Antissenso/genética , Oligonucleotídeos Antissenso/toxicidade , RNA Mensageiro/genética , Telomerase/antagonistas & inibidores , Telomerase/genética , Fatores de Tempo
12.
Sensors (Basel) ; 19(22)2019 Nov 12.
Artigo em Inglês | MEDLINE | ID: mdl-31726718

RESUMO

The data validity of safe driving in the Internet of Vehicles (IoV) is the basis of improving the safety of vehicles. Different from a traditional information systems, the data anomaly analysis of vehicle safety driving faces the diversity of data anomaly and the randomness and subjectivity of the driver's driving behavior. How to combine the characteristics of the IOV data with the driving style analysis to provide effective real-time anomaly data detection has become an important issue in the IOV applications. This paper aims at the critical safety data analysis, considering the large computing cost generated by the real-time anomaly detection of all data in the data package. We preprocess it through the traffic cellular automata model which is built to achieve the ideal abnormal detection effect with limited computing resources. On the basis of this model, the Anomaly Detection based on Driving style (ADD) algorithm is proposed to realize real-time and online detection of anomaly data related to safe driving. Firstly, this paper designs the driving coefficient and proposes a driving style quantization model to represent the driving style of the driver. Then, based on driving style quantization model and vehicle driving state information, a data anomaly detection algorithm is developed by using Gaussian mixture model (GMM). Finally, combining with the application scenarios of multi-vehicle collaboration in the Internet of Vehicles, this paper uses real data sets and simulation data sets to analyze the effectiveness of the proposed ADD algorithm.

13.
Int J Mol Sci ; 20(2)2019 Jan 16.
Artigo em Inglês | MEDLINE | ID: mdl-30654562

RESUMO

Flax (Linum usitatissimum L.) is an important industrial crop that is often cultivated on marginal lands, where salt stress negatively affects yield and quality. High-throughput RNA sequencing (RNA-seq) using the powerful Illumina platform was employed for transcript analysis and gene discovery to reveal flax response mechanisms to salt stress. After cDNA libraries were constructed from flax exposed to water (negative control) or salt (100 mM NaCl) for 12 h, 24 h or 48 h, transcription expression profiles and cDNA sequences representing expressed mRNA were obtained. A total of 431,808,502 clean reads were assembled to form 75,961 unigenes. After ruling out short-length and low-quality sequences, 33,774 differentially expressed unigenes (DEUs) were identified between salt-stressed and unstressed control (C) flax. Of these DEUs, 3669, 8882 and 21,223 unigenes were obtained from flax exposed to salt for 12 h (N1), 24 h (N2) and 48 h (N4), respectively. Gene function classification and pathway assignments of 2842 DEUs were obtained by comparing unigene sequences to information within public data repositories. qRT-PCR of selected DEUs was used to validate flax cDNA libraries generated for various durations of salt exposure. Based on transcriptome sequences, 1777 EST-SSRs were identified of which trinucleotide and dinucleotide repeat microsatellite motifs were most abundant. The flax DEUs and EST-SSRs identified here will serve as a powerful resource to better understand flax response mechanisms to salt exposure for development of more salt-tolerant varieties of flax.


Assuntos
Linho/genética , Perfilação da Expressão Gênica , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Análise de Sequência de RNA/métodos , Cloreto de Sódio/toxicidade , Estresse Fisiológico/genética , Análise por Conglomerados , Regulação para Baixo/efeitos dos fármacos , Regulação para Baixo/genética , Etiquetas de Sequências Expressas , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Genes de Plantas , Repetições de Microssatélites/genética , Anotação de Sequência Molecular , Reprodutibilidade dos Testes , Estresse Fisiológico/efeitos dos fármacos , Transcriptoma/genética , Regulação para Cima/efeitos dos fármacos , Regulação para Cima/genética
14.
Adv Physiol Educ ; 42(3): 449-453, 2018 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-29972058

RESUMO

Peer instruction has been used extensively in lecture courses; however, there is little evidence of its use in laboratory courses. The purpose of the present study was to describe the implementation of the peer instruction method in a physiology laboratory course in China. Second-year medical students attended a 6-wk physiology laboratory course in the fall semester of the 2016-2017 school year. In the six new physiology laboratory classes, peer instruction strategies were used to substitute for the traditional short, didactic lectures. The effects of peer instruction were measured by in-class quizzes and confidence levels. The students' evaluations of peer instruction were measured by a Likert scale questionnaire. Peer instruction significantly improved the mean score on quizzes (0.53 ± 0.50 vs. 0.68 ± 0.47, P < 0.001) and confidence levels (2.36 ± 0.66 vs. 2.80 ± 0.45, P < 0.001). Furthermore, for individual incorrect answers, 39.07% changed to correct answers after peer instruction, whereas, for correct answers, 6.61% were changed to an incorrect response. Overall, significantly more students changed their answers from incorrect to correct than from correct to incorrect [χ2: 333.11; degrees of freedom (df): 1; P < 0.001]. Therefore, the positive effects of peer instruction were higher than the negative effects (χ2: 244.55; df: 1; P < 0.001). Moreover, student evaluations of peer instruction were highly positive. In conclusion, the implementation of peer instruction to the physiology laboratory course is an effective strategy to enhance students' performance on in-class quizzes and confidence levels. In addition, the attitude of students toward peer instruction was favorable.


Assuntos
Currículo , Avaliação Educacional/métodos , Grupo Associado , Fisiologia/educação , Estudantes de Medicina , China , Currículo/normas , Avaliação Educacional/normas , Humanos
15.
Environ Monit Assess ; 190(7): 391, 2018 Jun 11.
Artigo em Inglês | MEDLINE | ID: mdl-29892786

RESUMO

This study used a total of 474 groundwater samples analyzed from 2014 data to evaluate the distribution of groundwater quality in the Water and Sanitation Agency (WASA) jurisdiction of Lahore city, Pakistan. The study further assessed the variations in suitability of groundwater for drinking (emphasis on arsenic and fluoride) and irrigation using spatial correlation technique in GIS. The hydrochemical analysis revealed a predominance of Mg-Ca-HCO3-SO4 and Ca-Mg-HCO3-SO4 type. Distribution analysis indicated relatively higher salinity (TDSmax = 1667 mg/L), total hardness (THmax = 558 mg/L), and alkalinity (HCO3-max = 584 mg/L) in the south-eastern region of the city, while the central part displayed the highest levels of SO4 and NO3. Also, the eastern region (north-south) of Lahore had significantly elevated As concentrations (up to 86 µg/L). The order of exceedance in terms of arsenic was Gunj Bakhsh town (17.4%), Nishter town (16.4%), Iqbal town (9.8%), Aziz Batti and Shalimar town (8.1%), and Ravi town (3%). The groundwater was classified as average saline to highly saline, except few samples in Aziz Batti/Shalimar town that were in non-saline group. Otherwise, the various indices classified the groundwater for irrigation as generally acceptable. With the various irrigation quality indices displaying discernible variations for the entire study area, it was observed from the distribution maps that the groundwater suitability for irrigation is relatively excellent in the areas away from industries and landfill locations. Also, the chloride analysis shows 98.7% of the groundwater samples belong to the very fresh and fresh water class. Thus, continued monitoring and studying the changes in groundwater quality in Lahore is imperative.


Assuntos
Água Potável/análise , Monitoramento Ambiental/métodos , Água Subterrânea/análise , Poluentes Químicos da Água/análise , Abastecimento de Água , Irrigação Agrícola/métodos , Arsênio/análise , Água Potável/química , Fluoretos/análise , Água Doce/análise , Água Subterrânea/química , Paquistão , Fosfatos , Salinidade , Análise Espacial , Qualidade da Água/normas
16.
Cancer Sci ; 108(1): 12-22, 2017 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-27862769

RESUMO

The molecular mechanism underlying non-alcoholic fatty liver disease progression to hepatocellular carcinoma (HCC) remains unknown. In this study, immunohistochemistry staining results showed that NS5ABP37 protein, which is in a state of lower expression in tumor tissues, decreased with increasing degree of HCC malignancy. Two cell models, HepG2 and L02, were used to analyze the mechanism between NS5ABP37 and HCC. In agreement, NS5ABP37 protein overexpression significantly suppressed cell proliferation, caused G1 /S cell cycle arrest, and induced apoptosis by increasing caspase-3/7 activity and cleaved caspase-3 levels. In addition, NS5ABP37 overexpression resulted in decreased intracellular triglyceride and total cholesterol contents, with level reduction in sterol regulatory element-binding proteins (SREBPs) and downstream effectors. Furthermore, NS5ABP37 overexpression decreased SREBP1c and SREBP2 levels by reducing their respective promoters. Finally, reactive oxygen species levels and endoplasmic reticulum stress were both induced by NS5ABP37 overexpression. These findings together indicate that NS5ABP37 inhibits cancer cell proliferation and promotes apoptosis, by altering SREBP-dependent lipogenesis and cholesterogenesis in HepG2 and L02 cells and inducing oxidative stress and endoplasmic reticulum stress.


Assuntos
Carcinoma Hepatocelular/metabolismo , Colesterol/biossíntese , Fibronectinas/metabolismo , Lipogênese , Neoplasias Hepáticas/metabolismo , Apoptose , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/patologia , Caspase 3/metabolismo , Caspase 7/metabolismo , Linhagem Celular Tumoral , Proliferação de Células , Progressão da Doença , Estresse do Retículo Endoplasmático , Fibronectinas/deficiência , Fibronectinas/genética , Humanos , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/patologia , Estadiamento de Neoplasias , RNA Mensageiro/análise , RNA Mensageiro/genética , Espécies Reativas de Oxigênio/metabolismo , Proteínas de Ligação a Elemento Regulador de Esterol/metabolismo , Triglicerídeos/metabolismo
17.
Tumour Biol ; 39(2): 1010428317694313, 2017 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-28240051

RESUMO

Studies have demonstrated that microRNA 185 may be a promising therapeutic target in liver cancer. However, its role in hepatocellular carcinoma is largely unknown. In this study, the proliferation of human HepG2 cells was inhibited by transfection of microRNA 185 mimics. Cell-cycle analysis revealed arrest at the G0/G1 phase. Transfection of HepG2 cells with microRNA 185 mimics significantly induced apoptosis. These data confirmed microRNA 185 as a potent cancer suppressor. We demonstrated that microRNA 185 was a compelling inducer of autophagy, for the first time. When cell autophagy was inhibited by chloroquine or 3-methyladenine, microRNA 185 induced more cell apoptosis. MicroRNA 185 acted as a cancer suppressor by regulating AKT1 expression and phosphorylation. Dual-luciferase reporter assays indicated that microRNA 185 suppressed the expression of target genes including RHEB, RICTOR, and AKT1 by directly interacting with their 3'-untranslated regions. Binding site mutations eliminated microRNA 185 responsiveness. Our findings demonstrate a new role of microRNA 185 as a key regulator of hepatocellular carcinoma via autophagy by dysregulation of AKT1 pathway.


Assuntos
Carcinoma Hepatocelular/genética , Neoplasias Hepáticas/genética , MicroRNAs/genética , Proteínas Proto-Oncogênicas c-akt/genética , Autofagia/genética , Materiais Biomiméticos/administração & dosagem , Carcinoma Hepatocelular/metabolismo , Carcinoma Hepatocelular/patologia , Carcinoma Hepatocelular/terapia , Ciclo Celular/genética , Processos de Crescimento Celular/genética , Terapia Genética/métodos , Células Hep G2 , Humanos , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/patologia , Neoplasias Hepáticas/terapia , MicroRNAs/administração & dosagem , MicroRNAs/biossíntese , Proteínas Proto-Oncogênicas c-akt/metabolismo , Transdução de Sinais , Transfecção/métodos
19.
Mol Vis ; 20: 1434-42, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25352749

RESUMO

PURPOSE: We conducted a meta-analysis aiming to evaluate the relationship between a common polymorphism (rs2511989 G>A) in the SERPING1 gene and the risk of age-related macular degeneration (AMD). METHODS: The PubMed, CISCOM, CINAHL, Web of Science, Google Scholar, EBSCO, Cochrane Library, and CBM databases were searched for relevant articles published before November 1, 2013, without any language restrictions. A meta-analysis was conducted using STATA 12.0 software. We calculated a crude odds ratio (OR) with a 95% confidence interval (95% CI) to evaluate the relationships under five genetic models. RESULTS: Seven case-control studies with a total of 7,159 patients with AMD and 5,797 healthy subjects met the inclusion criteria. The results of our meta-analysis showed that the SERPING1 rs2511989 polymorphism might be correlated with an increased risk of AMD (G allele versus A allele: OR = 1.09, 95% CI = 1.03-1.15, p = 0.020; GG + GA versus AA: OR = 1.14, 95% CI = 1.03-1.26, p = 0.014; GG versus GA+AA: OR = 1.10, 95% CI = 1.02-1.19, p = 0.012; GG versus AA: OR = 1.20, 95% CI = 1.07-1.34, p = 0.002; respectively). Results of subgroup analysis by ethnicity revealed positive correlations between the SERPING1 rs2511989 polymorphism and risk of AMD among Caucasians under five genetic models (all p<0.05), but not among Asians (all p>0.05). CONCLUSIONS: The current meta-analysis shows that the SERPING1 rs2511989 polymorphism may have a positive effect on the risk of AMD, especially among Caucasians.


Assuntos
Proteínas Inativadoras do Complemento 1/genética , Predisposição Genética para Doença , Degeneração Macular/genética , Polimorfismo de Nucleotídeo Único , Idoso , Idoso de 80 Anos ou mais , Alelos , Povo Asiático , Estudos de Casos e Controles , Proteína Inibidora do Complemento C1 , Feminino , Humanos , Degeneração Macular/etnologia , Degeneração Macular/patologia , Masculino , Pessoa de Meia-Idade , Modelos Genéticos , Razão de Chances , Fatores de Risco , População Branca
20.
Electrophoresis ; 35(23): 3345-54, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-25224837

RESUMO

The latest developments in chiral analysis of ß-blocker drugs by capillary electromigration techniques are reviewed in this article. Following the previous review by Aturki et al. [Electrophoresis 2011, 32, 2602-2628], this review includes the papers published during the period from January 2011 to December 2013. During this time, some novel chiral selectors were reported and applied to improve the enantioseparation of ß-blocker drugs and structurally related compounds. These chiral selectors include CDs and their derivatives, macrocyclic antibiotics, tartrate complexs, the monolithic molecularly imprinted polymer, and the polymeric surfactants. In addition, this article summarizes the methodological improvements for enhancing sensitivity in chiral analysis of ß-blockers and structurally related compounds by CE. The involved authors described the use of online sample preconcentration techniques to increase the detection sensitivity in the enantiomeric analysis of a broad range of samples.


Assuntos
Antagonistas Adrenérgicos beta/análise , Antagonistas Adrenérgicos beta/química , Eletroforese Capilar/métodos , Sensibilidade e Especificidade , Estereoisomerismo
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