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1.
Materials (Basel) ; 13(10)2020 May 14.
Artigo em Inglês | MEDLINE | ID: mdl-32422987

RESUMO

The load capacity of reinforced concrete structure will decrease by chloride ingress in coastal region. In this paper, the corrosion probability and flexural strength of a typical reinforced concrete beam design under the influence of temperature and humidity was obtained by the Monte Carlo method. The relationship between flexural strength, temperature, relative humidity, and geometric parameters was established. The annual average temperature and relative humidity were treated as random variables together with the geometric size and concrete compressive strength. The corrosion probability and flexural strength in a wave splashing zone, coastal atmospheric zone, and offshore atmospheric zone were calculated. The results show that the corrosion probabilities of the three regions are obviously different. When the standard deviation of temperature is less than 1.5 °C, the temperature can be treated as a constant in the calculation of the concrete cracking probability in the wave splashing zone. A binary logistic regression formula was given to predict whether the randomness of temperature and humidity should be considered in the offshore atmospheric zone. When the standard deviation of the temperature is less than 1 °C, the temperature randomness has no significant effect on the flexural strength of beams in the wave splashing zone. The flexural strength distribution conforms to the normal distribution in the early stage of service and the Weibull distribution after concrete cracking.

2.
Ying Yong Sheng Tai Xue Bao ; 28(2): 367-374, 2017 Feb.
Artigo em Chinês | MEDLINE | ID: mdl-29749142

RESUMO

In order to reveal the dynamics of canopy vertical structure and its effects on understory regeneration, we built 24 permanent plots (20 m×20 m) on the upslope, midslopeand downslope, respectively, in a typical evergreen broadleaved forest in Damingshan, Guangxi, China. We measured the crown area of each tree with diameter at breast height (DBH)≥1.0 cm, and surveyed the understory regeneration in growing season from 2009 to 2011. The results showed that the total canopy cover significantly increased from 54.0% in 2009 to 67.4% in 2011 after the frozen disaster in 2008. A significant difference existed in the cover and increment of different canopy layers. The canopy cover in the upper layers was markedly higher than that in the middle and lower layers. The increment of canopy coverage in the middle and lower layers was significantly higher than that in the upper layer. There were 55 regenerated woody plant species, and the dominant families and species of regenerated plants were in accord with those in the evergreen broadleaved forest. Biodiversity index of regenerated plants in the same slope position was significantly different among different years, and no significant difference was observed among different slope positions in the same year. The correlation between the coverage at different canopy layers and the species richness and abundance of regenerated plants was not significant. Total canopy cover and canopy coverage at the middle and lower layers were significantly negatively correlated with the Shannon index, Simpson index, and Pielou evenness index of the understory regenerated plants. It indicated that canopy coverage had a significant influence on the regeneration of understory, and the middle and lower layers had a stronger influence on the biodiversity of regenerated plants.


Assuntos
Ecossistema , Florestas , Biodiversidade , China , Árvores
3.
Pathol Oncol Res ; 22(2): 393-9, 2016 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-26581611

RESUMO

The hepatitis B virus X protein (HBx), which is encoded by hepatitis B virus (HBV), plays crucial roles in the tumorigenesis of HBV associated hepatocellular carcinoma (HCC). Recent studies suggest that the HBx is involved in regulation of host immune cytokines and chemokines in HBV-associated HCC patients. However, effects of the HBx on autocrine chemokine expression profiles of hepatoma cells, which were shown in modulation of tumor-immune cell interactions, have not been investigated comprehensively. In the present study, human hepatoma cell lines SMMC-7721 and HepG2 were transfected with HBx-expressing plasmid. Human chemokine antibody array 1 (RayBio®), which simultaneously detects 38 chemokine factors, was used to determine chemokine expression profiles. Real-time polymerase chain reaction (real-time PCR) was used to further confirm the differential expression of chemokines. Chemokine antibody array revealed that all 38 chomekines were found to be expressed by SMMC-7721 and HepG2 cell lines. Interleukin-8 (IL-8) was obviously up-regulated, and epithelial neutrophil-activating protein 78 (ENA78), eosinophil chemotactic protein-1 (Eotaxin-1), monocyte chemotactic protein-1 (MCP-1), MCP-2, MCP-3 and macrophage inflammatory protein-3ß (MIP-3ß) were significantly declined in both cell lines following transfection of HBx-expressing plasmid. Other chemokines showed little or no significant changes. HBx-induced differential chemokine expression levels were validated by real-time PCR. Hierarchical cluster analysis identified a distinction of chomekine expression profiles between HBX-expressing hepatoma cell lines and controls. Our findings provide new evidence that HBx is able to selectively regulate chomekines in hepatoma cells that may be involved in the regulation of tumor-immune cell interactions.


Assuntos
Carcinoma Hepatocelular/metabolismo , Quimiocinas/metabolismo , Vírus da Hepatite B/fisiologia , Hepatite B/metabolismo , Neoplasias Hepáticas/metabolismo , Transativadores/metabolismo , Western Blotting , Carcinoma Hepatocelular/patologia , Carcinoma Hepatocelular/virologia , Quimiocinas/genética , Hepatite B/patologia , Hepatite B/virologia , Humanos , Técnicas Imunoenzimáticas , Neoplasias Hepáticas/patologia , Neoplasias Hepáticas/virologia , Análise Serial de Proteínas , RNA Mensageiro/genética , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transativadores/genética , Células Tumorais Cultivadas , Proteínas Virais Reguladoras e Acessórias
4.
Exp Mol Pathol ; 99(3): 399-408, 2015 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-26341139

RESUMO

Our previous study showed hepatitis B virus X protein (HBx) suppresses the p16 expression in hepatocarcinogenesis. In this study we explored the relationship between HBx and trimethylation of H3K9 (H3K9me3), and elucidated the underlying mechanisms in HBx inducing the tumor suppressor p16 gene silence. SMMC-7721 and HepG2 hepatoma cell lines were transfected with HBx-expressing plasmid. Immunohistochemistry, Western blotting and real-time polymerase chain reaction, were performed to detect the expressions of HBx, H3K9me3, and jumonji domain-containing protein 2B (JMJd2B). H3K9me3 enrichment on the p16 promoter was measured by immunoprecipitation-PCR (ChIP-PCR) analyses, and 39 cases of hepatitis B virus (HBV) associated-hepatocellular carcinoma (HCC) and corresponding noncancerous liver tissues were also examined. We demonstrated that HBx was able to upregulate H3K9me3 and suppress JMJd2B mRNA and protein levels in SMMC-7721 and HepG2 hepatoma cell lines. JMJd2B, as a specific target of H3K9me3 for demethylation, was inversely correlated with the levels of H3K9me3 in SMMC-7721 (r=-0.666, P<0.05) and HepG2 cells (r=-0.625, P<0.05). The ChIP-PCR data indicated that HBx remarkably increased H3K9me3 on the p16 promoter region. Immunohistochemistry analysis showed that H3K9me3 expression in HBx positive HCC samples were significantly higher than that in HBx negative HCC tissues and were associated with decreased levels of JMJd2B expression. JMJd2B immunoreactivity was also remarkably inversed to that of HBx in HCC tissues (r=-0.630, P<0.05). Our results provide evidence that HBx is able to induce H3K9me3 on the p16 promoter via the decrease of demethylase JMJd2B expression and thus promote the repression of p16 gene expression to enhance hepatocarcinogenesis.


Assuntos
Carcinoma Hepatocelular/metabolismo , Transformação Celular Neoplásica , Genes p16 , Vírus da Hepatite B/genética , Histonas/metabolismo , Neoplasias Hepáticas/metabolismo , Transativadores/genética , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/patologia , Carcinoma Hepatocelular/virologia , Linhagem Celular Tumoral , Proliferação de Células/genética , Humanos , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/patologia , Neoplasias Hepáticas/virologia , Lisina/metabolismo , Metilação , Regiões Promotoras Genéticas , Proteínas Virais Reguladoras e Acessórias
5.
Chin J Integr Med ; 21(5): 376-83, 2015 May.
Artigo em Inglês | MEDLINE | ID: mdl-25519444

RESUMO

OBJECTIVE: To investigate the preventive effects of Qiangzhi Decoction (, QZD) on influenza A pneumonia through inhibition of inflammatory cytokine storm in vivo and in vitro. METHODS: One hundred ICR mice were randomly divided into the virus control, the Tamiflu control and the QZD high-, medium-, and low-dose groups. Mice were infected intranasally with influenza virus (H1N1) at 10 median lethal dose (LD50). QZD and Tamiflu were administered intragastrically twice daily from day 0 to day 7 after infection. The virus control group was treated with distilled water alone under the same condition. The number of surviving mice was recorded daily for 14 days after viral infection. The histological damage and viral replication and the expression of inflammatory cytokines were monitored. Additionally, the suppression capacity on the secretion of regulated on activation normal T cells expressed and secreted (RANTES) and tumor necrosis factor-α (TNF-α) in epithelial and macrophage cell-lines were evaluated. RESULTS: Compared with the virus control group, the survival rate of the QZD groups significantly improved in a dose-dependent manner (P<0.05), the viral titers in lung tissue was inhibited (P<0.05), and the production of inflammatory cytokines interferon-γ (IFN-γ), interleukin-6 (IL-6), TNF-α, and intercellular adhesion molecule-1 (ICAM-1) were suppressed (P<0.05). Meanwhile, the secretion of RANTETS and TNF-α by epithelial and macrophage cell-lines was inhibited with the treatment of QZD respectively in vitro (p<0.05) CONCLUSIONS: The preventive effects of QZD on influenza virus infection might be due to its unique cytokine inhibition mechanism. QZD may have significant therapeutic potential in combination with antiviral drugs.


Assuntos
Citocinas/metabolismo , Medicamentos de Ervas Chinesas/uso terapêutico , Inflamação/patologia , Vírus da Influenza A Subtipo H1N1/fisiologia , Infecções por Orthomyxoviridae/prevenção & controle , Pneumonia/prevenção & controle , Substâncias Protetoras/uso terapêutico , Animais , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Quimiocina CCL5/metabolismo , Quimiocinas/metabolismo , Cães , Medicamentos de Ervas Chinesas/farmacologia , Ensaio de Imunoadsorção Enzimática , Hemaglutinação por Vírus/efeitos dos fármacos , Humanos , Vírus da Influenza A Subtipo H1N1/efeitos dos fármacos , Vírus da Influenza A Subtipo H1N2/efeitos dos fármacos , Pulmão/efeitos dos fármacos , Pulmão/patologia , Células Madin Darby de Rim Canino , Camundongos Endogâmicos ICR , Infecções por Orthomyxoviridae/complicações , Infecções por Orthomyxoviridae/patologia , Pneumonia/complicações , Pneumonia/patologia , Substâncias Protetoras/farmacologia , Taxa de Sobrevida , Fator de Necrose Tumoral alfa/farmacologia
6.
J Cardiothorac Surg ; 8: 146, 2013 Jun 07.
Artigo em Inglês | MEDLINE | ID: mdl-23758949

RESUMO

Glomus tumor is usually a small, benign tumor and typically occurs in the dermis or subcutis or soft tissue of the extremities and rarely in the visceral locations. Its occurrence in the main bronchus is extremely rare. The current case reported a 30-year-old woman with dyspnea on exertion and hemoptysis, she had a glomus tumor which has large size, deep location and exhibits an infiltrative margin as well as increased atypical mitotic figures. These characteristics suggest malignant behavior. However, there is little data regarding glomus tumors arising in the bronchus, the need for caution in diagnosing this case as a malignant glomus tumor must be highlighted. Therefore, the diagnosis of bronchial glomus tumor of uncertain malignant potential was favored. To the best of our knowledge, both the type and the location of this glomus tumor are extremely rare. Accumulation of more cases are needed to clarify their diagnosis and significance since there is little data regarding glomus tumors arising in the bronchus.


Assuntos
Neoplasias Brônquicas/diagnóstico , Neoplasias Brônquicas/cirurgia , Tumor Glômico/diagnóstico , Tumor Glômico/cirurgia , Adulto , Biópsia , Neoplasias Brônquicas/patologia , Broncoscopia , Diagnóstico Diferencial , Diagnóstico por Imagem , Feminino , Tumor Glômico/patologia , Humanos , Excisão de Linfonodo , Invasividade Neoplásica
7.
Exp Mol Pathol ; 95(2): 144-50, 2013 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-23797005

RESUMO

The aim of the present study was to optimize the pregelatinized starch technique for cell block preparation and apply this approach in cultured cells of all types of growing forms, suspension and adherent. In order to evenly mix the starch powder and the cell suspension, we crafted a special plastic dropper. To prove the effectiveness of this optimized technique we used different cell lines, NCI-H69, NCI-H345, HCT-116, SKBR3 and MDA-MB-231. The morphology features, immunocytochemistry (ICC) and fluorescent/chromogenic in-situ hybridization (FISH/CISH) on the cell block sections were evaluated. The morphology features, the ICC and ISH results of cell block sections prepared by the new method were satisfactory comparing with the results obtained in biopsies, the gold standard test for this kind of analysis. The most attractive advantage of our optimized pregelatinized starch technique is that this new method is based on cell suspensions instead of cell sediment, so with our technique every section will contain cells due to the even distribution of the starch powder and the cells forming a homogeneous cell block. To the authors' knowledge, this is the first description on cell block preparation based on cell suspension.


Assuntos
Técnicas de Preparação Histocitológica , Linhagem Celular , Gelatina , Humanos , Amido
8.
J Cardiothorac Surg ; 8: 131, 2013 May 17.
Artigo em Inglês | MEDLINE | ID: mdl-23684153

RESUMO

We herein reported a primary pulmonary papillary carcinoma with colloid-like luminal content in the glandular cavity and classic nuclear features such as pseudo-inclusions, intranuclear grooves in the tumor cell nuclei and ground glass nuclei which closely mimics papillary thyroid carcinoma. Meanwhile, lymph node in the left pulmonary hilum was involved and showed similar features to the primary pulmonary papillary carcinoma. This specific histopathological presentation caused a diagnostic dilemma.The patient didn't show previous concomitant or subsequent evidence of a thyroid tumor. Immunohistochemistry further confirmed pulmonary origin and excluded a metastasis from the thyroid, as it was thyroglobulin negative, thyroid transcription factor 1 and surfactant apoprotein A positive, which was consistent with the imageology and history.Based on the above features, the diagnosis of primary pulmonary papillary carcinoma was confirmed. Understanding the existence of papillary thyroid carcinoma-like pulmonary papillary carcinoma will avoid misdiagnosis or unnecessary clinical and radiologic investigations in future.


Assuntos
Adenocarcinoma/diagnóstico por imagem , Adenocarcinoma/cirurgia , Neoplasias Pulmonares/diagnóstico por imagem , Neoplasias Pulmonares/cirurgia , Adenocarcinoma/patologia , Idoso de 80 Anos ou mais , Carcinoma/diagnóstico por imagem , Carcinoma Papilar , Diagnóstico Diferencial , Evolução Fatal , Hemorroidectomia , Humanos , Imuno-Histoquímica , Neoplasias Pulmonares/patologia , Excisão de Linfonodo , Masculino , Pneumonectomia , Radiografia , Câncer Papilífero da Tireoide , Neoplasias da Glândula Tireoide/diagnóstico por imagem , Ultrassonografia
9.
Ultrasound Med Biol ; 39(7): 1215-22, 2013 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-23602805

RESUMO

The purpose of this study was to investigate the diagnostic value of shear wave elasticity imaging (SWEI) and real-time elastography (RTE) in liver fibrosis induced by dimethylnitrosamine (DMN) and to compare the accuracy of these methods. Seventy male Wistar rats given a single intra-peritoneal injection of DMN and 10 control rats given a saline injection underwent SWEI and RTE to determine their shear wave velocity (V(s)) and liver fibrosis (LF) index, respectively. Correlations between V(s) or the LF index and histologic stage of liver fibrosis (S0-S4) were analyzed, and the diagnostic values of the techniques were assessed using a receiver operating characteristic curve. A positive correlation was found between V(s) and stage of liver fibrosis (r = 0.947, p < 0.001) and between LF index and stage (S) of liver fibrosis (r = 0.662, p < 0.001). For Vs, the areas under the receiver operating characteristic curve for the diagnosis of fibrosis, S ≥ S1, S ≥ S2, S ≥ S3 and S = S4, were 0.983, 0.995, 0.999 and 0.964, respectively; for the LF index, the values were 0.871, 0.887, 0.761 and 0.839, respectively (all p < 0.001). Vs and the LF index values in rats with severe inflammatory activity were significantly higher than those in controls (p < 0.001). In conclusion, positive correlations exist between V(s) or the LF index and the severity of liver fibrosis in rats. Vs is more accurate than the LF index in predicting liver fibrosis in rats. However, severe inflammatory activity may reduce the accuracy of both techniques.


Assuntos
Modelos Animais de Doenças , Técnicas de Imagem por Elasticidade/métodos , Interpretação de Imagem Assistida por Computador/métodos , Cirrose Hepática/diagnóstico por imagem , Animais , Humanos , Aumento da Imagem/métodos , Ratos , Ratos Wistar , Reprodutibilidade dos Testes , Sensibilidade e Especificidade
10.
Am J Respir Crit Care Med ; 186(11): 1160-7, 2012 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-23043084

RESUMO

RATIONALE: Effective treatment for lung cancer requires accuracy in subclassification of carcinoma subtypes. OBJECTIVES: To identify microRNAs in bronchial brushing specimens for discriminating small cell lung cancer (SCLC) from non-small cell lung cancer (NSCLC) and for further differentiating squamous cell carcinoma (SQ) from adenocarcinoma (AC). METHODS: Microarrays were used to screen 723 microRNAs in laser-captured, microdissected cancer cells from 82 snap-frozen surgical lung specimens. Quantitative reverse-transcriptase polymerase chain reaction was performed on 153 macrodissected formalin-fixed, paraffin-embedded (FFPE) surgical lung specimens to evaluate seven microRNA candidates discovered from microarrays. Two microRNA panels were constructed on the basis of a training cohort (n = 85) and validated using an independent cohort (n = 68). The microRNA panels were applied as differentiators of SCLC from NSCLC and of SQ from AC in 207 bronchial brushing specimens. MEASUREMENTS AND MAIN RESULTS: Two microRNA panels yielded high diagnostic accuracy in discriminating SCLC from NSCLC (miR-29a and miR-375; area under the curve [AUC], 0.991 and 0.982 for training and validation data set, respectively) and in differentiating SQ from AC (miR-205 and miR-34a; AUC, 0.977 and 0.982 for training and validation data set, respectively) in FFPE surgical lung specimens. Moreover, the microRNA panels accurately differentiated SCLC from NSCLC (AUC, 0.947) and SQ from AC (AUC, 0.962) in bronchial brushing specimens. CONCLUSIONS: We found two microRNA panels that accurately discriminated between the three subtypes of lung carcinoma in bronchial brushing specimens. The identified microRNA panels may have considerable clinical value in differential diagnosis and optimizing treatment strategies based on lung cancer subtypes.


Assuntos
Carcinoma Pulmonar de Células não Pequenas/patologia , Carcinoma de Células Escamosas/patologia , Neoplasias Pulmonares/patologia , MicroRNAs/genética , Carcinoma de Pequenas Células do Pulmão/patologia , Idoso , Lavagem Broncoalveolar/métodos , Carcinoma Pulmonar de Células não Pequenas/genética , Carcinoma Pulmonar de Células não Pequenas/cirurgia , Carcinoma de Células Escamosas/genética , Carcinoma de Células Escamosas/cirurgia , Linhagem Celular Tumoral , Distribuição de Qui-Quadrado , Diagnóstico Diferencial , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Modelos Lineares , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/cirurgia , Masculino , Pessoa de Meia-Idade , Inclusão em Parafina , Curva ROC , Reação em Cadeia da Polimerase em Tempo Real/métodos , Reprodutibilidade dos Testes , Estudos de Amostragem , Carcinoma de Pequenas Células do Pulmão/genética , Carcinoma de Pequenas Células do Pulmão/cirurgia
11.
Zhonghua Gan Zang Bing Za Zhi ; 20(5): 386-9, 2012 May.
Artigo em Chinês | MEDLINE | ID: mdl-22971287

RESUMO

OBJECTIVE: To investigate the feasibility of real-time elastography for quantitative evaluation of liver fibrosis in a rat model. METHODS: A total of 70 male Wistar rats were included in the group for dimethylnitrosamine (DMN)-induced liver injury, and 10 saline-injected rats were used as normal control. Hepatic injury was induced by a single intraperitoneal injection of DMN at a dose of 50 mg/kg of body weight. Nine or ten rats in the group with DNM injected and one or two rats in the normal control group were randomly selected and sacrificed at each of the following post-injection time: day 5, 7, 10, 14, 21, 24, and 28. And their livers were taken for pathology analysis. All the rats underwent real-time elastography before sacrificed in order to acquire area ratio of low-strain region (% AREA) and liver fibrosis index (LF index) which were compared with the stage of liver fibrosis and grade of necroinflammatory pathologically. By the different data, Spearman correlation analysis, rank-sum test or receiver operating characteristic curve was used. RESULTS: Among 58 successfully modeled rats, there were nine, 13, 14 and 12 rats of S1, S2, S3 and S4 liver fibrosis on pathology, respectively, which were with or without mild necroinflammatory. The other 10 rats were found to be S0 with severe necroinflammatory. Values of LF index and % AREA both increased with liver fibrosis stage (P less than 0.05). There was certain correlation between LF index and liver fibrosis stage (r=0.643, P=0.000), so was % AREA and liver fibrosis stage (r=0.662, P=0.000). As for LF index, Areas under the receiver operating characteristic curve (Az) was 0.943, 0.890, 0.743 and 0.821 for the diagnosis of hepatic fibrosis S1 or higher, S2 or higher, S3 or higher and S4, respectively; as for % AREA, they were 0.948, 0.883, 0.772 and 0.842, respectively. However, we found a significant difference for LF index or % AREA between S0 with and without severe inflammatory activity rats (P=0.005 and P=0.017). CONCLUSION: Real-time elastography is available for quantitative assessment of liver fibrosis in rats induced by DMN, but severe inflammatory activity can affect its accuracy.


Assuntos
Técnicas de Imagem por Elasticidade , Cirrose Hepática Experimental/patologia , Fígado/patologia , Animais , Dimetilnitrosamina/efeitos adversos , Cirrose Hepática Experimental/induzido quimicamente , Masculino , Ratos , Ratos Wistar
12.
Acta Virol ; 56(2): 101-10, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22720699

RESUMO

UNLABELLED: Previously, we have found a new mutation at nt 1726-1730 that is associated with lower hepatitis B virus (HBV) DNA levels in the liver, and mutations at nt 1762/1764 that are correlated with higher HBV DNA levels. To confirm the effects of these mutations on the virus replication efficiency, substitutions nt 1726-1730 CTGAG and A1762T/G1764A in the HBV X (HBX) gene region were investigated alone or in combination. Cells Huh-7 or HepG2 were transfected with these constructs. The effects of these mutations on HBV were investigated at the gene and protein levels. The double mutation A1762T/G1764A increased whereas the nt 1726-1730 CTGAG mutations decreased the levels of released virion-associated and intracellular HBV DNA. The combined mutations had no appreciable effect on the replication capacity of the virus. Cells bearing the constructs with double mutations A1762T/G1764A contained the lowest levels of hepatitis B e antigen (HBeAg). Lowest expression of HBV X protein was in constructs that had both A1762T/G1764A and 1726-1730 CTGAG mutations. We think that changes in secondary RNA structure that were caused by these mutations might have been responsible for those results. KEYWORDS: hepatitis B virus; X gene; mutants; replication.


Assuntos
Vírus da Hepatite B , Regiões Promotoras Genéticas , Antígenos E da Hepatite B , Vírus da Hepatite B/genética , Humanos , Mutação , Replicação Viral
13.
Huan Jing Ke Xue ; 33(2): 652-7, 2012 Feb.
Artigo em Chinês | MEDLINE | ID: mdl-22509611

RESUMO

A solid-phase extraction (SPE)-high performance liquid chromatography (HPLC) method has been developed for simultaneous determination of 10 sulfonamide antibiotics in water. The analytes were first enriched and purified through a PEP solid-phase extraction column, and eluted with acetonitrile-dichloromethane solution (2: 1, V/V), then detected by a HPLC with a UV detector. The detection wavelength was 268 nm and the column temperature was 33 degrees C, using gradient elution process with acetonitrile - 0.4% acetic acid/water (V/V) as the mobile phase to achieve baseline separations of these 10 analytes. The linearity range was 10 - 2 000 microg x L(-1). The recovery ranges of standard addition for deionized water and real water samples were 73.4% - 95.6% and 70.2% - 92.5%, respectively (except for sulfonamide, were 8.5% and 8.0%). The limit of detection was 1.42-7.25 ng x L(-1). Application of this method for parts of Huangpu River in Shanghai, surface water and groundwater in Chongming Island showed that sulfonamide antibiotics were detected in different frequencies in different aqueous environments, with the concentration range of 13.3 - 241.5 ng x L(-1), proving this method is easy, rapid, sensitive and efficient to meet the needs of actual work.


Assuntos
Antibacterianos/análise , Cromatografia Líquida de Alta Pressão/métodos , Sulfonamidas/análise , Poluentes Químicos da Água/análise , Antibacterianos/classificação , China , Água Subterrânea/análise , Rios , Sensibilidade e Especificidade , Extração em Fase Sólida , Sulfonamidas/classificação
14.
Waste Manag ; 32(1): 137-43, 2012 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-21978423

RESUMO

The effect of ammonia inhibition was evaluated during the enhanced anaerobic treatment of digested effluent from a 700m(3) chicken-manure continuous stirred tank reactor (CSTR). A 12.3L internal circulation (IC) reactor inoculated with an anaerobic granular sludge and operated at 35±1°C was employed for the investigation. With a corresponding organic loading rate of 1.5-3.5kg-COD/m(3)d over a hydraulic retention time of 1.5d, a maximum volumetric biogas production rate of 1.2m(3)/m(3)d and TCOD (total COD) removal efficiency ranging from 70% to 80% was achieved. However, the continual increase in the influent TAN content led to ammonia inhibition in the methanogenesis system. The SCOD/TAN (soluble COD/total ammonia nitrogen) ratio was presented to be the key controlling factor for the anaerobic treatment of semi-digested chicken manure, and further validation through shock loading and ammonia inhibition experiments was conducted. The threshold value of the SCOD/TAN ratio was determined to be 2.4 (corresponding to a TAN of 1250mg/L) at an influent pH of 8.5-9.


Assuntos
Amônia/farmacologia , Reatores Biológicos , Fermentação/efeitos dos fármacos , Esterco , Animais , Galinhas
17.
Histol Histopathol ; 26(1): 23-35, 2011 01.
Artigo em Inglês | MEDLINE | ID: mdl-21117024

RESUMO

A remarkable feature of HBV-associated HCC is male predominance. The cooperation of hepatitis B virus X protein (HBx) with androgen receptor (AR) signaling pathway has been documented to contribute to this dominance. HBx, a multifunctional viral regulator, has been documented to induce promoter hypermethylation and low expression of tumor suppressor genes via activation of DNA methyl-transferase (DNMT) in hepatocarcinogenesis. In prostate cancer, hypermethylation of AR promoter is associated with loss of AR expression. However, the relationship among HBx, DNMTs, the methylation status of AR and AR expression in HBV-associated HCC is still unknown. In this report, we found that HBx correlated with high levels of AR in HCC cases and induced AR expression by stimulating its transcription in liver cell lines. HBx correlated with high expression of DNMTs in HCC cases too. Both in vivo and in vitro, however, the expression of AR was not associated with its promoter methylation status, and the methylation status of AR was not regulated by DNMTs. AR expression is higher in peritumoral tissues than in tumors, as well as being higher in HBV-associated HCC than in HBV-negative cases. Therefore, HBx-induced high expression of AR plays a role during hepatocarcinogenesis, but is not involved with its promoter methylation or DNMTs. HBx-mediated DNMT deregulation is gene-specific, and the expression and methylated regulation of AR is tissue-specific.


Assuntos
Carcinoma Hepatocelular/etiologia , Hepatite B/complicações , Neoplasias Hepáticas/etiologia , Receptores Androgênicos/genética , Receptores Androgênicos/metabolismo , Transativadores/metabolismo , Adulto , Idoso , Sequência de Bases , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/metabolismo , Carcinoma Hepatocelular/virologia , Linhagem Celular Tumoral , DNA (Citosina-5-)-Metiltransferases/metabolismo , Metilação de DNA , Primers do DNA/genética , Feminino , Hepatite B/genética , Hepatite B/metabolismo , Hepatite B/virologia , Vírus da Hepatite B/metabolismo , Vírus da Hepatite B/patogenicidade , Humanos , Técnicas In Vitro , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/virologia , Masculino , Pessoa de Meia-Idade , Regiões Promotoras Genéticas , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , RNA Neoplásico/genética , RNA Neoplásico/metabolismo , Transdução de Sinais , Proteínas Virais Reguladoras e Acessórias
18.
Exp Mol Pathol ; 89(3): 268-75, 2010 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-20620135

RESUMO

The hepatitis B virus×protein (HBx) has been implicated as a potential trigger of the epigenetic deregulation of some genes, but the underlying mechanism remains unknown. The aim of this study is to identify underlying mechanisms involved in HBx-mediated epigenetic modification in the process of HBx induced p16(INK4A) promoter hypermethylation. Liver cell lines were stably transfected with HBx-expressing vector. The methylation status of p16(INK4A) was examined by methyl-specific polymerase chain reaction (MSP) and bisulfite sequencing. Reverse transcription and real-time polymerase chain reaction (real-time RT-PCR), Western blot and immunohistochemistry were used to analyze the expression of HBx, HBx-mediated DNA methylation abnormalities and p16(INK4A). Some cases of HCC and corresponding noncancerous liver tissues were studied. HBx up-regulates DNMT1 and DNMT3A expression in both mRNA level and protein level, and HBx represses p16(INK4A) expression through inducing hypermethylation of p16(INK4A) promoter. Moreover, HBx induces hypermethylation of p16(INK4A) promoter through DNMT1 and DNMT3A. Regulation of DNMT1 and DNMT3A by HBx promoted hypermethylation of p16(INK4A) promoter region. HBx-DNMTs-p16(INK4A) promoter hypermethylation may suggest a mechanism for tumorigenesis during hepatocarcinogenesis.


Assuntos
Carcinoma Hepatocelular/virologia , DNA (Citosina-5-)-Metiltransferases/biossíntese , Regulação Neoplásica da Expressão Gênica , Genes p16 , Neoplasias Hepáticas/virologia , Transativadores/metabolismo , Adulto , Idoso , Western Blotting , Carcinoma Hepatocelular/enzimologia , Carcinoma Hepatocelular/genética , Linhagem Celular Tumoral , DNA (Citosina-5-)-Metiltransferase 1 , Metilação de DNA/genética , DNA Metiltransferase 3A , Feminino , Humanos , Imuno-Histoquímica , Neoplasias Hepáticas/enzimologia , Neoplasias Hepáticas/genética , Masculino , Pessoa de Meia-Idade , Regiões Promotoras Genéticas/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transfecção , Regulação para Cima , Proteínas Virais Reguladoras e Acessórias
19.
J Cancer Res Clin Oncol ; 136(8): 1255-65, 2010 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-20182889

RESUMO

PURPOSE: Krüppel-like factor 8 (KLF8) plays an important role in oncogenic transformation and is highly overexpressed in several types of human cancer. We investigated the expression of KLF8 in renal cell carcinoma (RCC) tissues and the role of small interference RNA targeting KLF8 on growth, cell cycle, and apoptosis of human renal carcinoma cell line 786-0 in vitro and in vivo. METHODS: The expression of KLF8 protein and mRNA in human renal carcinoma samples was detected by immunochemistry and reverse transcription polymerase chain reaction (RT-PCR). The effects of small interference RNA (siRNA) targeting KLF8 on growth, invasiveness, cell cycle, and apoptosis of 786-0 cells were evaluated by MTT assay, Matrigel Invasion Assay, and flow cytometry in vitro. We also investigated effect of siRNA targeting KLF8 on growth of 786-0 cells in nude mice in vivo. RESULTS: Immunohistochemistry and RT-PCR results showed the expression of KLF8 protein and mRNA in RCC specimens was significantly higher than that in the adjacent non-tumorous renal tissues (P < 0.001). KLF8-siRNA depressed the cellular growth and invasion of 786-0 cells in vitro. The flow cytometry results revealed that KLF8-siRNA could induce an increase in G0/G1 phase cells and induce cell apoptosis. Intratumor injection of siRNA targeting KLF8 inhibited the growth of 786-0 cells in vivo in nude mice tumor model. CONCLUSIONS: KLF8 possibly involved in regulating the cell growth, invasion, apoptosis, and proliferation of renal carcinoma cancer cells. Blocking the KLF8 channel might be a potential therapeutic strategy for RCC.


Assuntos
Neoplasias Renais/genética , RNA Interferente Pequeno/genética , Proteínas Repressoras/genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Animais , Sequência de Bases , Biomarcadores Tumorais/genética , Divisão Celular , Linhagem Celular Tumoral , Primers do DNA , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Imuno-Histoquímica , Neoplasias Renais/patologia , Fatores de Transcrição Kruppel-Like , Masculino , Camundongos , Camundongos Nus , Pessoa de Meia-Idade , RNA Mensageiro/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transfecção
20.
J Surg Res ; 159(1): 443-50, 2010 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-20031158

RESUMO

The gelatinases B (MMP9) and A (MMP2) are two members of the matrix metalloproteinase (MMPs) family that are expressed in human cancer, and play a critical role in tumor cell invasion and metastasis. Caveolin-1 (Cav1) has recently been identified as a tumor metastasis modifier gene. However, the effect and mechanism of Cav1 in pancreatic carcinoma cell invasion remain unknown. In this study, we investigated the expression of Cav1, MMP2, and MMP9 in several different pancreatic carcinoma cell lines. We transfected pcDNA3.0-Cav1 plasmid and Cav1 siRNA into SW1990 and Bxpc3 cells, respectively. Using cell invasion assay, we found that overexpression of Cav1 inhibited cell invasion, whereas the knockdown of Cav1 in Bxpc3 cells promoted cell invasion. Moreover, to explore the mechanisms underlying these observations, we further investigated the expression of MMP2, MMP9, phospho-Akt, and phospho-Erk by Western blot, and the activities of MMP2 and MMP9 by gelatin zymography. The results indicated that Cav1 gene could inhibit pancreatic carcinoma cell invasion, at least in part, probably through Erk-MMP signal pathway, suggesting that the endogenous expression or re-expression of Cav1 might help therapeutically reduce their invasive potential in pancreatic carcinoma cells.


Assuntos
Carcinoma/enzimologia , Caveolina 1/metabolismo , Metaloproteinase 2 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/metabolismo , Neoplasias Pancreáticas/enzimologia , Caveolina 1/genética , Linhagem Celular Tumoral , Cromonas , Células Epiteliais/enzimologia , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Flavonoides , Técnicas de Silenciamento de Genes , Humanos , Sistema de Sinalização das MAP Quinases , Quinases de Proteína Quinase Ativadas por Mitógeno/antagonistas & inibidores , Morfolinas , Invasividade Neoplásica , Ductos Pancreáticos/enzimologia , Inibidores de Fosfoinositídeo-3 Quinase , Proteínas Proto-Oncogênicas c-akt/metabolismo , Interferência de RNA , RNA Mensageiro/metabolismo , RNA Interferente Pequeno/metabolismo
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