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1.
Zhongguo Zhong Yao Za Zhi ; 46(7): 1592-1597, 2021 Apr.
Artigo em Chinês | MEDLINE | ID: mdl-33982456

RESUMO

For the field detection problems of critical quality attribute(CQA) of moisture content in traditional Chinese medicine(TCM) manufacturing process, big brand TCM Tongren Niuhuang Qingxin Pills were used as the carrier, to establish a moisture content NIR field detection model with or without cellophane in real world production with use of near infrared(NIR) spectroscopy combined with stoichiometry. With the moisture content determined by drying method as reference value, the partial least square method(PLS) was used to analyze the correlation between the spectrum and the moisture reference value. Then the spectral pretreatment methods were screened and optimized to further improve the accuracy and stability of the model. The results showed that the best quantitative model was developed by the spectral data pretreatment of standard normal variate(SNV) with the latent variable factor number of 2 and 7 of Tongren Niuhuang Qingxin Pills with or without cellophane samples. The prediction coefficient of determination(R_(pre)~2) and standard deviation of prediction(RMSEP) of the model with cellophane samples were 0.765 7 and 0.157 2%; R_(pre)~2 and RMSEP of the model without cellophane samples were 0.772 2 and 0.207 8%. The NIR quantitative models of moisture content of Tongren Niuhuang Qingxin Pills with and without cellophane both showed good predictive performance to realize the rapid, accurate and non-destructive quantitative analysis of moisture content in such pills, and provide a method for the field quality control of the critical chemical attributes of moisture in the manufacturing of big brand TCM.


Assuntos
Medicamentos de Ervas Chinesas , Medicina Tradicional Chinesa , Análise dos Mínimos Quadrados , Espectroscopia de Luz Próxima ao Infravermelho
2.
Zhongguo Zhong Yao Za Zhi ; 46(7): 1598-1605, 2021 Apr.
Artigo em Chinês | MEDLINE | ID: mdl-33982457

RESUMO

Texture sensory attributes are the key items in quality control of Chinese medicinal honeyed pills. The purpose of this study is to develop a quality control method for assessing the texture sensory attributes of Chinese medicinal honeyed pills based on real-world Tongren Niuhuang Qingxin pilular masses and finished products. First, parameters of texture profile analysis(TPA) were optimized through single factor and central composite design(CCD) experiments to establish a detection method for texture sensory attri-butes of Tongren Niuhuang Qingxin Pills. The results showed that the established detection method was stable and reliable, with the optimal parameters set up as follows: deformation percentage of 70%, detection speed at 30 mm·min~(-1), and interval time of 15 s. Furthermore, 540 data points yielded form six texture sensory attributes of pills from 30 batches were subjected to multivariate statistical process control(MSPC) with Hotelling T~2 and squared prediction error(SPE) control charts to establish the quality control method of Tongren Niuhuang Qingxin Pills. This study is expected to provide a reference for improving the quality control system of Chinese medicinal honeyed pills.


Assuntos
Medicamentos de Ervas Chinesas , Medicina Tradicional Chinesa , Controle de Qualidade
3.
Artigo em Inglês | MEDLINE | ID: mdl-32994137

RESUMO

BACKGROUND: Group A Streptococcus (GAS) is an important pathogen causing morbidity and mortality worldwide. Surveillance of resistance and emm type has important implication to provide helpful information on the changing GAS epidemiology and empirical treatment. METHODS: To study the emergence of resistant GAS in children with upper respiratory tract infection (URTI), a retrospective study was conducted from 2000 to 2019 in southern Taiwan. Microbiological studies, including antibiotic susceptibility, were performed. GAS emm types and sequences were determined by molecular methods. The population was divided into two separate decades to analyze potential changes over time. The 1st decade was 2000-2009; the 2nd decade was 2010-2019. Multivariate analyses were performed to identify independent risk factors associated with macrolide resistance between these periods. RESULTS: A total of 320 GAS from 339 children were enrolled. Most of the children (75%) were under 9 years of age. The most common diagnosis was scarlet fever (225, 66.4%), and the frequency increased from 54.8% in the 1st to 77.9% in the 2nd decade (p < 0.0001). There was a significant increase in resistance to erythromycin and azithromycin from 18.1%, 19.3% in the 1st to 58.4%, 61.0% in the 2nd decade (p < 0.0001). This was associated with clonal expansion of the GAS emm12-ST36 which carrying erm(B) and tet(M) from 3.0% in the 1st to 53.2% in the 2nd decade (p < 0.0001). CONCLUSIONS: Significant emergence of macrolide-resistant GAS emm12-ST36 in children supports the need for continuing surveillance and investigation for the clonal virulence.

4.
Front Microbiol ; 11: 117, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32117141

RESUMO

Group A streptococcus (GAS) is a versatile pathogen that causes a wide spectrum of diseases in humans. Invading host cells is a known strategy for GAS to avoid antibiotic killing and immune recognition. However, the underlying mechanisms of GAS resistance to intracellular killing need to be explored. Endothelial HMEC-1 cells were infected with GAS, methicillin-resistant Staphylococcus aureus (MRSA) and Salmonella Typhimurium under nicotinamide (NAM)-supplemented conditions. The intracellular NAD+ level and cell viability were respectively measured by NAD+ quantification kit and protease-based cytotoxicity assay. Moreover, the intracellular bacteria were analyzed by colony-forming assay, transmission electron microscopy, and confocal microscopy. We found that supplementation with exogenous nicotinamide during infection significantly inhibited the growth of intracellular GAS in endothelial cells. Moreover, the NAD+ content and NAD+/NADH ratio of GAS-infected endothelial cells were dramatically increased, whereas the cell cytotoxicity was decreased by exogenous nicotinamide treatment. After knockdown of the autophagy-related ATG9A, the intracellular bacterial load was increased in nicotinamide-treated endothelial cells. The results of Western blot and transmission electron microscopy also revealed that cells treated with nicotinamide can increase autophagy-associated LC3 conversion and double-membrane formation during GAS infection. Confocal microscopy images further showed that more GAS-containing vacuoles were colocalized with lysosome under nicotinamide-supplemented conditions than without nicotinamide treatment. In contrast to GAS, supplementation with exogenous nicotinamide did not effectively inhibit the growth of MRSA or S. Typhimurium in endothelial cells. These results indicate that intracellular NAD+ homeostasis is crucial for controlling intracellular GAS infection in endothelial cells. In addition, nicotinamide may be a potential new therapeutic agent to overcome persistent infections of GAS.

5.
Front Microbiol ; 9: 1733, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30123194

RESUMO

Group A Streptococcus (GAS) is a human pathogen causing a wide spectrum of diseases, from mild pharyngitis to life-threatening necrotizing fasciitis. GAS has been shown to evade host immune killing by invading host cells. However, how GAS resists intracellular killing by endothelial cells is still unclear. In this study, we found that strains NZ131 and A20 have higher activities of NADase and intracellular multiplication than strain SF370 in human endothelial cells (HMEC-1). Moreover, nga mutants of NZ131 (SW957 and SW976) were generated to demonstrate that NADase activity is required for the intracellular growth of GAS in endothelial cells. We also found that intracellular levels of NAD+ and the NAD+/NADH ratio of NZ131-infected HMEC-1 cells were both lower than in cells infected by the nga mutant. Although both NZ131 and its nga mutant were trapped by LC3-positive vacuoles, only nga mutant vacuoles were highly co-localized with acidified lysosomes. On the other hand, intracellular multiplication of the nga mutant was increased by bafilomycin A1 treatment. These results indicate that NADase causes intracellular NAD+ imbalance and impairs acidification of autophagosomes to escape autophagocytic killing and enhance multiplication of GAS in endothelial cells.

6.
Sci Rep ; 7(1): 5676, 2017 07 18.
Artigo em Inglês | MEDLINE | ID: mdl-28720835

RESUMO

Annexin A2 (ANXA2), a phospholipid-binding protein, has multiple biological functions depending on its cellular localization. We previously demonstrated that IFN-γ-triggered ANXA2 secretion is associated with exosomal release. Here, we show that IFN-γ-induced autophagy is essential for the extracellular secretion of ANXA2 in lung epithelial cells. We observed colocalization of ANXA2-containing autophagosomes with multivesicular bodies (MVBs) after IFN-γ stimulation, followed by exosomal release. IFN-γ-induced exophagic release of ANXA2 could not be observed in ATG5-silenced or mutant RAB11-expressing cells. Furthermore, knockdown of RAB8A and RAB27A, but not RAB27B, reduced IFN-γ-triggered ANXA2 secretion. Surface translocation of ANXA2 enhanced efferocytosis by epithelial cells, and inhibition of different exophagic steps, including autophagosome formation, fusion of autophagosomes with MVBs, and fusion of amphisomes with plasma membrane, reduced ANXA2-mediated efferocytosis. Our data reveal a novel route of IFN-γ-induced exophagy of ANXA2.


Assuntos
Anexina A2/metabolismo , Autofagia/fisiologia , Interferon gama/farmacologia , Pulmão/efeitos dos fármacos , Células A549 , Células Epiteliais/efeitos dos fármacos , Células HEK293 , Humanos , Células Jurkat , Corpos Multivesiculares , Fagocitose , Proteínas rab de Ligação ao GTP/metabolismo , Proteínas rab27 de Ligação ao GTP/metabolismo
7.
Int J Mol Med ; 39(6): 1564-1570, 2017 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-28487934

RESUMO

Late diagnosis and lack of specific therapeutic targets contribute to the low survival rate of patients with epithelial ovarian cancer (EOC), the most lethal gynecologic malignancy. Therefore, the screening of diagnostic markers and the identification of therapeutic targets are urgently required. Heat shock factor 1 (HSF1) has been demonstrated to be overexpressed in certain malignancies and to be involved in tumor initiation, development, transformation and metastasis. It is believed that HSF1 is a promising candidate for antitumor therapy. However, its expression pattern and function in ovarian cancer are far from being fully elucidated. Therefore, we examined the HSF1 expression in human EOC tissues, and evaluated its carcinogenesis-promoting activity in a xenograft tumor model. Examination of HSF1 expression in human EOC tissues was performed by immunohistochemical assay using ovarian tissue blots. Specific short hairpin RNA (shRNA) against HSF1 was employed to knockdown HSF1 in SKOV3 cells. Cell proliferative activity was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay; cell cycle distribution and apoptosis were determined by flow cytometric analysis. In normal ovarian tissues, HSF1 was barely detected, whereas, high expression of HSF1 was found in malignant EOC tissues, including serous, mucinous, endometrioid, and clear cell EOC tissues. Suppressed proliferative activity and intensified apoptosis were observed in HSF1-knockdown SKOV3 cells. In nude mouse xenografts, downregulation of HSF1 was found to cause reduced carinogenesis, indicating the antitumor effect induced by modulation of HSF1 against EOC. Our findings suggest that HSF1 may be considered as a potential candidate diagnostic marker of human EOC, and that modulation of HSF1 could be a promising therapeutic strategy against human EOC.


Assuntos
Carcinogênese/genética , Regulação Neoplásica da Expressão Gênica , Fatores de Transcrição de Choque Térmico/genética , Neoplasias Epiteliais e Glandulares/genética , Neoplasias Ovarianas/genética , Animais , Apoptose , Carcinogênese/patologia , Carcinoma Epitelial do Ovário , Linhagem Celular Tumoral , Proliferação de Células , Feminino , Fatores de Transcrição de Choque Térmico/análise , Humanos , Camundongos Nus , Neoplasias Epiteliais e Glandulares/patologia , Neoplasias Ovarianas/patologia , Ovário/metabolismo , Ovário/patologia , Interferência de RNA , RNA Interferente Pequeno/genética
8.
Huan Jing Ke Xue ; 38(12): 5169-5173, 2017 Dec 08.
Artigo em Chinês | MEDLINE | ID: mdl-29964578

RESUMO

This study used short process nitrification sludge to investigate the effect of high free ammonia (FA) on ammonia escape during ammonia oxidation. A sequencing batch reactor (SBR) was used to study the ammonia escape process under different FA concentrations. The results indicate that FA in water is usually combined with water molecules at lower FA concentrations to produce stable NH3·H2O with almost no ammonia escape. With high FA concentration (FA>687.1 mg·L-1), ammonia nitrogen is not oxidized to oxidized nitrogen (NO2--N concentration is <0.1 mg·L-1 at the end of aeration), but the total nitrogen (TN) reached 269.7 mg·L-1. Therefore, when there is a high FA concentration, NH4+-N will evaporate in the form of FA, resulting in ammonia escape. In addition, when 226.6 ≤ FA ≤ 711.8 mg·L-1, the free ammonia escape rate (FEV) grows rapidly with the increase in FA concentration.


Assuntos
Amônia/química , Reatores Biológicos , Nitrificação , Esgotos , Nitrogênio/química , Oxirredução , Eliminação de Resíduos Líquidos
9.
Oncotarget ; 7(48): 78946-78957, 2016 Nov 29.
Artigo em Inglês | MEDLINE | ID: mdl-27793047

RESUMO

S100A4 is a calcium-binding protein capable of promoting epithelial-mesenchymal transition. Previously, we have demonstrated that S100A4 is required to sustain the head and neck cancer-initiating cells (HN-CICs) subpopulation. In this study, to further investigate the molecular mechanism, we established the head and neck squamous cell carcinoma (HNSCC) cell lines stably expressing mutant S100A4 proteins with defective calcium-binding sites on either N-terminal (NM) or C-terminal (CM), or a deletion of the last 15 amino-acid residues (CD). We showed that the NM, CM and CD harboring sphere cells that were enriched with HN-CICs population exhibited impaired stemness and malignant properties in vitro, as well as reduced tumor growth ability in vivo. Mechanistically, we demonstrated that mutant S100A4 proteins decreased the promoter activity of Nanog, likely through inhibition of p53. Moreover, the biophysical analyses of purified recombinant mutant S100A4 proteins suggest that both NM and CM mutant S100A4 were very similar to the WT S100A4 with subtle difference on the secondary structure, and that the CD mutant protein displayed the unexpected monomeric form in the solution phase.Taken together, our results suggest that both the calcium-binding ability and the C-terminal region of S100A4 are important for HN-CICs to sustain its stemness property and malignancy, and that the mechanism could be mediated by repressing p53 and subsequently activating the Nanog expression.


Assuntos
Cálcio/metabolismo , Carcinoma de Células Escamosas/metabolismo , Neoplasias de Cabeça e Pescoço/metabolismo , Células-Tronco Neoplásicas/metabolismo , Proteína A4 de Ligação a Cálcio da Família S100/metabolismo , Animais , Sítios de Ligação , Carcinoma de Células Escamosas/genética , Carcinoma de Células Escamosas/patologia , Linhagem Celular Tumoral , Proliferação de Células , Regulação Neoplásica da Expressão Gênica , Neoplasias de Cabeça e Pescoço/genética , Neoplasias de Cabeça e Pescoço/patologia , Humanos , Camundongos Endogâmicos BALB C , Camundongos Nus , Mutação , Proteína Homeobox Nanog/genética , Proteína Homeobox Nanog/metabolismo , Células-Tronco Neoplásicas/patologia , Fenótipo , Regiões Promotoras Genéticas , Domínios Proteicos , Estrutura Secundária de Proteína , Proteína A4 de Ligação a Cálcio da Família S100/química , Proteína A4 de Ligação a Cálcio da Família S100/genética , Transdução de Sinais , Carcinoma de Células Escamosas de Cabeça e Pescoço , Relação Estrutura-Atividade , Fatores de Tempo , Transfecção , Carga Tumoral , Proteína Supressora de Tumor p53/genética , Proteína Supressora de Tumor p53/metabolismo
10.
Int J Mol Sci ; 17(6)2016 Jun 14.
Artigo em Inglês | MEDLINE | ID: mdl-27314333

RESUMO

R-spondin 1 (Rspo1) plays an essential role in stem cell biology by potentiating Wnt signaling activity. Despite the fact that Rspo1 holds therapeutic potential for a number of diseases, its biogenesis is not fully elucidated. All Rspo proteins feature two amino-terminal furin-like repeats, which are responsible for Wnt signal potentiation, and a thrombospondin type 1 (TSR1) domain that can provide affinity towards heparan sulfate proteoglycans. Using chemical inhibitors, deglycosylase and site-directed mutagenesis, we found that human Rspo1 and Rspo3 are both N-glycosylated at N137, a site near the C-terminus of the furin repeat 2 domain, and Rspo2 is N-glycosylated at N160, a position near the N-terminus of TSR1 domain. Elimination of N-glycosylation at these sites affects their accumulation in media but have no effect on the ability towards heparin. Introduction of the N-glycosylation site to Rspo2 mutant at the position homologous to N137 in Rspo1 restored full glycosylation and rescued the accumulation defect of nonglycosylated Rspo2 mutant in media. Similar effect can be observed in the N137 Rspo1 or Rspo3 mutant engineered with Rspo2 N-glycosylation site. The results highlight the importance of N-glycosylation at these two positions in efficient folding and secretion of Rspo family. Finally, we further showed that human Rspo1 is subjected to endoplasmic reticulum (ER) quality control in N-glycan-dependent manner. While N-glycan of Rspo1 plays a role in its intracellular stability, it had little effect on secreted Rspo1. Our findings provide evidence for the critical role of N-glycosylation in the biogenesis of Rspo1.


Assuntos
Heparina/metabolismo , Processamento de Proteína Pós-Traducional , Via Secretória , Trombospondinas/metabolismo , Sítios de Ligação , Retículo Endoplasmático/metabolismo , Glicosilação , Células HEK293 , Humanos , Ligação Proteica , Dobramento de Proteína , Estabilidade Proteica , Trombospondinas/química
11.
Artigo em Inglês | MEDLINE | ID: mdl-24845451

RESUMO

The oriental sole Brachirus orientalis (Pleuronectiformes: Soleidae) is characterized by both eyes on the right side of the body and orbicular-ovate body. In this paper, the complete mitochondrial genome sequence of this sole was first determined. The total length is 16,602 bp, including 13 protein-coding genes, 22 tRNA genes, and 2 rRNA genes (12S and 16S), as well as a putative control region and a putative L-strand replication origin (OL). Gene contents, locations, and arrangements are identical to those of typical bony fishes. Overall base composition of the mitogenome is 30.4%, 28.6%, 15.3%, and 25.7% for A, C, G, and T, with a high A + T content (56.1%). The determination of B. orientalis complete mitogenome sequence could contribute to phylogenetic study on Soleidae and Pleuronectiformes.


Assuntos
DNA Mitocondrial/genética , Linguados/genética , Genoma Mitocondrial , Animais , Composição de Bases , Sequência de Bases/genética , Dados de Sequência Molecular , Filogenia , Análise de Sequência de DNA/veterinária
12.
J Clin Microbiol ; 54(1): 157-62, 2016 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-26560544

RESUMO

emm typing is the most widely used molecular typing method for the human pathogen Streptococcus pyogenes (group A streptococcus [GAS]). emm typing is based on a small variable region of the emm gene; however, the emm cluster typing system defines GAS types according to the nearly complete sequence of the emm gene. Therefore, emm cluster typing is considered to provide more information regarding the functional and structural properties of M proteins in different emm types of GAS. In the present study, 677 isolates collected between 1994 and 2008 in a hospital in southern Taiwan were analyzed by the emm cluster typing system. emm clusters A-C4, E1, E6, and A-C3 were the most prevalent emm cluster types and accounted for 67.4% of total isolates. emm clusters A-C4 and E1 were associated with noninvasive diseases, whereas E6 was significantly associated with both invasive and noninvasive manifestations. In addition, emm clusters D4, E2, and E3 were significantly associated with invasive manifestations. Furthermore, we found that the functional properties of M protein, including low fibrinogen-binding and high IgG-binding activities, were correlated significantly with invasive manifestations. In summary, the present study provides updated epidemiological information on GAS emm cluster types in southern Taiwan.


Assuntos
Antígenos de Bactérias/genética , Proteínas da Membrana Bacteriana Externa/genética , Proteínas de Transporte/genética , Variação Genética , Tipagem Molecular/métodos , Infecções Estreptocócicas/epidemiologia , Infecções Estreptocócicas/microbiologia , Streptococcus pyogenes/classificação , Streptococcus pyogenes/isolamento & purificação , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Criança , Pré-Escolar , Feminino , Fibrinogênio/metabolismo , Genótipo , Hospitais , Humanos , Imunoglobulina G/metabolismo , Lactente , Recém-Nascido , Masculino , Pessoa de Meia-Idade , Epidemiologia Molecular/métodos , Prevalência , Ligação Proteica , Streptococcus pyogenes/genética , Streptococcus pyogenes/patogenicidade , Taiwan/epidemiologia , Virulência , Adulto Jovem
13.
PLoS One ; 10(12): e0145223, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26710228

RESUMO

Clustered regularly interspaced short palindromic repeats (CRISPR) are the bacterial adaptive immune system against foreign nucleic acids. Given the variable nature of CRISPR, it could be a good marker for molecular epidemiology. Group A streptococcus is one of the major human pathogens. It has two CRISPR loci, including CRISPR01 and CRISPR02. The aim of this study was to analyze the distribution of CRISPR-associated gene cassettes (cas) and CRISPR arrays in highly prevalent emm types. The cas cassette and CRISPR array in two CRISPR loci were analyzed in a total of 332 strains, including emm1, emm3, emm4, emm12, and emm28 strains. The CRISPR type was defined by the spacer content of each CRISPR array. All strains had at least one cas cassette or CRISPR array. More than 90% of the spacers were found in one emm type, specifically. Comparing the consistency between emm and CRISPR types by Simpson's index of diversity and the adjusted Wallace coefficient, CRISPR01 type was concordant to emm type, and CRISPR02 showed unidirectional congruence to emm type, suggesting that at least for the majority of isolates causing infection in high income countries, the emm type can be inferred from CRISPR analysis, which can further discriminate isolates sharing the same emm type.


Assuntos
Antígenos de Bactérias/genética , Proteínas da Membrana Bacteriana Externa/genética , Proteínas de Transporte/genética , Repetições Palindrômicas Curtas Agrupadas e Regularmente Espaçadas/genética , DNA Intergênico/genética , Tipagem Molecular/métodos , Streptococcus pyogenes/genética , Humanos , Análise de Sequência de DNA
14.
BMC Genomics ; 16: 355, 2015 May 06.
Artigo em Inglês | MEDLINE | ID: mdl-25943439

RESUMO

BACKGROUND: The mitochondrial genomes (mitogenomes) of flatfishes (Pleuronectiformes) exhibit highly diversified types of large-scale gene rearrangements. We have reported that the mitogenomes of Crossorhombus azureus (Bothidae), Samariscus latus (Samaridae) and Cynoglossus fishes (Cynoglossidae) show different types of gene rearrangements. RESULTS: In the present study, the complete mitogenomes of two Symphurus species (Cynoglossidae), Symphurus plagiusa and Symphurus orientalis, were determined. The gene order in the S. plagiusa mitogenome is the same as that of a typical vertebrate (without any gene rearrangements). Surprisingly, large-scale gene rearrangements have occurred in S. orientalis. In the rearranged fragment from the control region (CR) to the WANCY tRNA cluster (tRNA cluster of tRNA-W, tRNA-A, tRNA-N, tRNA-C and tRNA-Y) in the S. orientalis mitogenome, tRNA-V and tRNA-M have been translocated to the 3' end of the 16S rRNA gene, with six large intergenic spacers over 20 bp in length. In addition, an origin for light-strand replication (OL) structure that is typically located in the WANCY region was absent in both the S. plagiusa and S. orientalis mitogenomes. It is generally recognized that a sequence in the WANCY region that encodes tRNAs forms a hairpin structure (OL-like structure) and can act as the OL when the typical locus is lost. Moreover, an additional OL-like structure was identified near the control region in the S. plagiusa mitogenome. CONCLUSIONS: The positions of the intergenic spacers and the rearranged genes of the S. orientalis mitogenome strongly indicate that the mechanism underlying the rearrangement of this mitogenome was Tandem Duplication and Random Loss. Additionally, two OL-like regions substituting for the typical locus were found in the S. plagiusa mitogenome. We speculate that the ancestral mitogenomes of S. plagiusa and S. orientalis also had this characteristic, such that if both OL-like structures functioned during mitochondrial replication, they could initiate duplicate replications of the light strand (L-strand), leading to duplication of the region between the two structures. We consider that this mechanism may account for the gene duplication that occurred during the gene rearrangement process in the evolution of the ancestral mitogenome to the S. orientalis mitogenome.


Assuntos
Linguados/genética , Genoma Mitocondrial , Animais , Duplicação Gênica , Rearranjo Gênico , Modelos Genéticos , Análise de Sequência de DNA
15.
Mitochondrial DNA ; 26(5): 732-3, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-24460167

RESUMO

Liachirus melanospilos (Pleuronectiformes: Soleidae) is characterized by both eyes on the right side of the body and pale brown color with black dots, small irregular, brown spots and large vague ocelli. In this study, the complete mitogenome sequence of this carpet sole has been determined. The total length is 17,001 bp, including 13 protein-coding genes, 22 tRNA genes and 2 rRNA genes (12S and 16S), as well as one control region and one L-strand replication origin (OL). Gene contents, locations and arrangements are identical to those of typical bony fishes. The nucleotide composition of the genome is 30.9%, 27.8%, 15.9% and 25.5% for A, C, G and T, respectively, with a slight bias of A + T content (56.4%). The determination of L. melanospilos complete mitogenome sequence could contribute to a better understanding of the systematic evolution of Soleidae and Pleuronectiformes.


Assuntos
Linguados/genética , Genoma Mitocondrial , Mitocôndrias/genética , Animais , Composição de Bases , Evolução Molecular , Linguados/fisiologia , Ordem dos Genes , Tamanho do Genoma , Análise de Sequência de DNA
16.
Mitochondrial DNA ; 26(1): 114-5, 2015 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-23789774

RESUMO

Aesopia cornuta belongs to the family Soleidae of Pleuronectiformes, and the morphological characters are much similar to those of Zebrias. In this article, we sequenced, characterized, and compared the complete mitogenome of A. cornuta for the first time. The genome is 16,737 base pairs in length, and is typically consist of 37 genes, including 13 protein-coding genes, two ribosomal RNA, 22 transfer RNA, as well as a putative L-strand replication origin and a putative control region. The gene organization is identical to that of typical bony fishes. The overall base composition is 29.1, 28.3, 26.8 and 15.8% for C, A, T and G, respectively, with a slight AT bias of 55.1%. This result is expected to contribute to understanding the systematic evolution of the genus Aesopia and further taxonomic and phylogenetic studies of Soleidae and Pleuronectiformes.


Assuntos
Linguados/genética , Genoma Mitocondrial , Mitocôndrias/genética , Animais , Composição de Bases , RNA Ribossômico/genética , RNA de Transferência/genética , Origem de Replicação , Análise de Sequência de DNA/métodos
17.
Guang Pu Xue Yu Guang Pu Fen Xi ; 34(7): 1758-62, 2014 Jul.
Artigo em Chinês | MEDLINE | ID: mdl-25269275

RESUMO

The vibrational levels of KH(X1 sigma+ v" = 0-3) were generated in the reaction of K(5P) with H2. The vibrationally excited KH(v" = 17) was populated by an overtone pump-probe configuration Different characteristics of collisional energy transfer in highly and lowly excited vibrational levels of KH and CO2 were investigated through measuring the time-resolved distribution of vibrational energy in KH(v" = 17.3) + CO2 collisions. For KH(v" = 17), there existed three principal regions of vibration temperature (T(v)) in this equilibration process. The initial phase consists of very rapid fall in T(v) within - 5 micros, and the vibrational energy of KH(v" = 17) is mainly transferred to the vibrational levels of CO2 (00 degrees 1) or high rotational levels of CO2 (00 degrees 0). The second phase (5-20 micros) has a slight decline in T(v), and the process of energy transfer to vibrational levels or high rotational levels of CO2 has already finished. The vibration temperature of the third phase has a slightly more rapid decline compared with the last phase. This phase shows that the process of transfer to lowly rotational levels and translation energy of CO2 is accelerated. The equilibration of vibrationally excited KH (v" = 3) in CO2 was also investigated. There are similarities to the behavior of KH (v" = 17) in CO2 plot, but also are significant differences. Once the initial resonant V-R exchange has equalized vibrational temperatures, there is a very slow linear decline in T(v) with equilibrium attained within -80 micros. This same point is reached within 15 micros for KH (v" = 17). The data demonstrate that single rate coefficient measurements are unlikely to capture the complex nature of processes that generally are multistaged with different relaxation rates characterizing each different stage. Examination of the quantum state distributions reveals that these distinct stages reflect the dominance of specific energy transfer mechanisms, some of which are inherently fast and others are much slower. The energy gain into CO2 resulting from collisions with excited KH was probed using transient absorption techniques. Distributions of nascent CO2 rotational populations in both ground (00 degrees 0) state and the vibrationally excited (00 degrees 1) state were determined. A kinetic model was developed to describe rate coefficients for appearance of CO2 states resulting from collisions with excited KH. These experiments show that collisions resulting in CO2 (00 degrees 0) are accompanied by substantial excitation in rotation while the vibrationally excited CO2 (00 degrees 1) state has rotational energy distributions near the initial distributions.

18.
Int J Mol Med ; 34(6): 1591-8, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-25318762

RESUMO

The difficulties faced in the effective treatment of ovarian cancer are multifactorial, but are mainly associated with relapse and drug resistance. Cancer stem-like cells have been reported to be an important contributor to these hindering factors. In this study, we aimed to investigate the anticancer activities of a bioactive fungal metabolite, namely terrein, against the human epithelial ovarian cancer cell line, SKOV3, primary human ovarian cancer cells and ovarian cancer stem-like cells. Terrein was separated and purified from the fermentation metabolites of the marine sponge-derived fungus, Aspergillus terreus strain PF26. Its anticancer activities against ovarian cancer cells were investigated by cell proliferation assay, cell migration assay, cell apoptosis and cell cycle assays. The ovarian cancer stem-like cells were enriched and cultured in a serum-free in vitro suspension system. Terrein inhibited the proliferation of the ovarian cancer cells by inducing G2/M phase cell cycle arrest. The underlying mechanisms involved the suppression of the expression of LIN28, an important marker gene of stemness in ovarian cancer stem cells. Of note, our study also demonstrated the ability of terrein to inhibit the proliferation of ovarian cancer stem-like cells, in which the expression of LIN28 was also downregulated. Our findings reveal that terrein (produced by fermention) may prove to be a promising drug candidate for the treatment of ovarian cancer by inhibiting the proliferation of cancer stem-like cells.


Assuntos
Aspergillus/química , Proliferação de Células/efeitos dos fármacos , Ciclopentanos/farmacologia , Pontos de Checagem da Fase G2 do Ciclo Celular/efeitos dos fármacos , Animais , Antineoplásicos/química , Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Apoptose/genética , Aspergillus/metabolismo , Western Blotting , Linhagem Celular Tumoral , Proliferação de Células/genética , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/genética , Ciclopentanos/química , Relação Dose-Resposta a Droga , Feminino , Pontos de Checagem da Fase G2 do Ciclo Celular/genética , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Biologia Marinha , Estrutura Molecular , Neoplasias Ovarianas/genética , Neoplasias Ovarianas/metabolismo , Neoplasias Ovarianas/patologia , Poríferos/microbiologia , Proteínas de Ligação a RNA/genética , Proteínas de Ligação a RNA/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Células Tumorais Cultivadas
19.
Huan Jing Ke Xue ; 35(7): 2658-63, 2014 Jul.
Artigo em Chinês | MEDLINE | ID: mdl-25244852

RESUMO

To improve the hydrolysis and acidification of activated sludge, two new ways, which were adding fermented sludge or sterilized fermented sludge to batch-mode activated sludge alkaline fermentation system, were investigated in this study. Temperature effect was also studied (10 degrees C and 35 degrees C). Result showed that mesophilic condition greatly enhanced sludge hydrolysis and acidification, which resulted in an obvious volatile fatty acids (VFAs) accumulation in the system. These two new ways could improve both sludge hydrolysis and acidification. The hydrolysis and acidification efficiencies of adding sterilized fermented sludge system were 2 and 1.5 times higher than those of adding fermented sludge system, respectively. In addition, VFAs content was nearly constant in a long time in sterilized fermented sludge addition system while it greatly consumed in fermented sludge addition system. The reason for this enhancement of two new ways were different, fermented sludge addition increased the amount of hydrolytic enzyme and acid-forming bacteria, while sterilized fermented sludge addition increased the amount of degradable substances, and the result also demonstrated that strengthen hydrolysis was more important than acidification. Therefore, mesophilic condition together with sterilized fermented sludge addition was a more effective way to enhance VFAs accumulation in alkaline sludge fermentation process.


Assuntos
Ácidos Graxos Voláteis/química , Fermentação , Esgotos/química , Bactérias , Reatores Biológicos , Concentração de Íons de Hidrogênio , Hidrólise
20.
Guang Pu Xue Yu Guang Pu Fen Xi ; 34(4): 884-7, 2014 Apr.
Artigo em Chinês | MEDLINE | ID: mdl-25007592

RESUMO

The radiative lifetimes and rate coefficients for deactivation of high lying 6(1)sigma+ state of NaK by collisions with H2 were studied. An OPO laser was set to a particular 2(1)sigma+ <-- 1(1)sigma+ transition. Another single mode Ti sapphire laser was then used to excite molecule from 2(1)sigma+ level to the 6(1)sigma+ state. The predissociation was monitored by the atomic potassium emission at the 3D --> 4P (1.7 microm) or the S --> 4P (1.24 microm), while bound state radiative processes were monitored by total fluorescence from the upper state to the various levels, all studied as a function of H2 density. The values for predissociation, collisional dissociation and collisional depopulation rate coefficients were obtained. The decay signal of the time resolved fluorescence from the 6(1)sigma+ --> 2(1)sigma+, 6(1)sigma+ -->1(1)sigma+ or 2(1)sigma+ --> 1(1)sigma+ transition was monitored. Based on the Stern-Volmer equation, the radiative lifetimes were monitored for 6(1)sigma+ --> 2(1)sigma+ and 2(1)sigma+ --> 1(1)sigma+ transition. The rate coefficients for deactivation of collisions with H2 were monitored for 6(1)sigma+ --> 2(1)sigma+, 6(1)sigma+ --> 1(1)sigma+ and 2(1)sigma+ -->1(1)sigma+. When the density of H2 was 10(19) cm(-3), the total collisional transfer energy (15 426 cm(-1)) and radiative energy (10 215 cm(-1)) were obtained. The relative fraction ((f(v)), (f(R)), (f(T)) of average energy disposal was derived as (0.58, 0.03, 0.39); (f(v)), (f(R)), (f(T)) represent separately the relative fraction of average energy disposal among vibration, rotation and translation. The major vibrational and translational energy release supports the assumption that the 6(1)sigma(+) -H2 collision occurs primarily in a collisional energy transfer mechanism. In this experiment, alkali molecules relative energy population ratio was determined through using the time integrated intensity, so we can get the total transfer energy. That the NaK (6(1)sigma+) energy transfers to the H2 vibrational, rotational and translational energy was quantitatively given for the first time, which illustrates the collisional mechanism.

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