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1.
Artigo em Inglês | MEDLINE | ID: mdl-32012968

RESUMO

Cemented paste backfill (CPB) is a common environmentally friendly mining approach. However, it remains undetermined whether CPB pollutes underground mine water. Tank leaching analysis of a CPB mass in distilled water was performed for 120 d, and water quality was tested in situ for a long-term pollution assessment. Computerized tomography was also used to determine the CPB micro-pore structure and ion-leaching mechanism. The dissolved Zn2+, Pb2+ and As5+ concentrations in the leachate peaked at 0.56, 0.11 and 0.066 mg/L, respectively, whereas the Co2+ and Cd2+ concentrations were lower than the detection limit. The CPB porosity decreased from 46.07% to 40.88% by soaking, and 80% of the pore diameters were less than 13.81 µm. The permeability decreased from 0.8 to 0.5 cm/s, and the quantity, length, and diameter of the permeate channels decreased with soaking. An in-situ survey showed novel selective solidification. The Zn2+ concentration in the mine water was 10-20 times that of the background water, and the Pb2+ concentration was 2-4 times the regulated value. Although the Pb2+ content decreased significantly with mining depth, there remains a serious environmental risk. Mine water pollution can be reduced by adding a solidifying agent for Pb2+ and Zn2+, during CPB preparation.

2.
Artigo em Inglês | MEDLINE | ID: mdl-31736338

RESUMO

Hepatocellular carcinoma (HCC) is the sixth common malignant tumor worldwide but current efficient and convenient screening methods remain lacking. This study aimed to discover a diagnostic or a screening biomarker from the urine of HBV-related HCC patients. We used iTRAQ coupled with mass spectrometry to identify candidate urinary proteins in a discovery cohort (n=40). The selected proteins were confirmed using ELISA in a validation cohort (n=140). Diagnostic performance of the selected proteins was assessed using receiver operating characteristic (ROC) and qualitative diagnostic analysis. A total of 96 differentially expressed proteins were identified. Urinary alpha-fetoprotein (u-AFP) and orosomucoid 1 (u-ORM1) were selected as target proteins by bioinformatics analysis and were significantly higher in HCC than in non-HCC patients as validated by western blot and ELISA. U-AFP had a strong correlation with serum AFP-L3 (Pearson r =0.944, p < 0.0001), indicating that u-AFP may be derived from circulating blood. The AUC of u-AFP was 0.795 with a sensitivity of 62.5% and a specificity of 95.4%, which showed no significantly difference with serum AFP (se-AFP). The AUC was 0.864 as u-AFP and u-ORM1 were combined, and performed much better than u-AFP or u-ORM1 alone. Qualitative diagnostic analysis showed that the positive predictive value of u-AFP was 90.1% and the diagnostic sensitivity of parallel combination of u-AFP and u-ORM1 was 85.1%. Taken together, AFP and ORM1 in the urine may be used as a diagnostic or screening biomarker of HCC and studies on large samples are needed to validate the result.

3.
Chemistry ; 25(68): 15586-15593, 2019 Dec 05.
Artigo em Inglês | MEDLINE | ID: mdl-31574171

RESUMO

Development of Pt group metal-free catalysts for low-temperature CO oxidation remains critical. In this work, active and stable mesoporous Cu-Ce-Ox solid solutions are prepared by using spray pyrolysis. The specific surface areas and pore volumes reach as high as 170 m2 g-1 and 0.24 cm3 g-1 , respectively. The results of CO oxidation study suggest that (1) the catalyst obtained by spray pyrolysis possesses much higher activity than those made by co-precipitation, sol-gel, and hydrothermal methods; (2) the optimal Cu0.2 -Ce0.8 -Ox solid solution presents a reactivity over 28 times that of both single-component CuO and CeO2 at 70 °C. Based on the study of pure-phase Cu-Ce-Ox solid solutions by selective leaching of segregated CuOx species, the active center for CO oxidation is confirmed as the bimetallic Cu-Ce-O site, whereas the individual CuOx particles not only act as spectators but also block the active Cu-Ce-O sites. A low apparent activation energy of approximately 48 kJ mol-1 is detected for CO oxidation at the Cu-Ce-O site, making Cu-Ce-Ox solid solutions able to present high activity at low temperature. Furthermore, the Cu-Ce-Ox catalysts exhibit excellent stability and thermal tolerance toward CO oxidation.

4.
Exp Ther Med ; 18(2): 1417-1425, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31316628

RESUMO

There are two main types of drugs that are used to treat chronic hepatitis B (CHB), including interferon (IFN) and nucleotide analogues. IFN inhibits the virus through direct antiviral activity and via immune regulation, and it has been widely applied for the treatment of CHB and other infections. However, the efficiency of IFN therapy is not entirely satisfactory. The aim of the present study was to investigate the factors affecting IFN therapy. The plasma of patients with CHB treated with IFN was collected and divided into the virological response group and non-virological response group according to their virological response. Serum proteins of virologically responsive patients were compared before and after IFN therapy using isobaric tags for relative and absolute quantitation technology. ELISA was used to validate these results in the same sample. In in vitro cell experiments, HepG2.2.15 cells were transfected with haptoglobin (Hp)-targeting small interfering RNA to inhibit expression of the Hp protein, and reverse transcription-quantitative polymerase chain reaction and western blotting were utilized to detect hepatitis B virus (HBV)-DNA, IFN and downstream molecular changes in the cell supernatants. The Hp protein levels were demonstrated to be significantly lower following 48 weeks of IFN therapy, and the levels of Hp in patients in the virological response group were significantly lower than those in the non-virological response group. In in vitro cell experiments, following inhibition of Hp protein expression, significantly decreased levels of HBV-DNA, and elevated levels of IFN and its downstream molecules were observed. These findings suggest that Hp may be able to predict the efficacy of IFN therapy, and it may inhibit HBV clearance. There is an association between Hp and IFN, which requires further clinical and laboratory studies to explore.

5.
Environ Technol ; : 1-12, 2019 Jan 29.
Artigo em Inglês | MEDLINE | ID: mdl-30694117

RESUMO

In this work, we developed a novel magnetic bimetallic Al/Fe (oxyhydr)oxide adsorbent through a facile and cost-effective method and explored its potential to adsorb fluoride in water. Its synthesis involved corrosion of natural magnetite in aluminium chloride solution, followed by titration with NaOH solution for in-situ synthesis of Al/Fe (oxyhydr)oxide-coated magnetite (Mag@Al2Fe). Characterization data indicated a uniform coating of Al/Fe (oxyhydr)oxide on magnetite, and the resulting composite possessed large specific surface area (∼90 m2/g) and good magnetic property. In batch adsorption experiments, the isotherm and kinetic data fitted well to the Langmuir model and pseudo-second-order model, respectively. The maximum adsorption capacity of Mag@Al2Fe is 26.5 mg/g, which was much higher than natural magnetite (0.44 mg/g). Moreover, this material retained high adsorption capacity toward fluoride within a wide pH range (3.0-8.0) and offered facile magnetic separation from water. Influence of competing ions was also evaluated which showed that the presence of Cl- and NO3- posed negligible interference, while HCO3- and SO42- had negative effects on fluoride adsorption. Thermodynamic investigations revealed that fluoride adsorption was exothermic and spontaneous. The observed increase in solution pH and formation of Al-F and Fe-F bonds (as indicated by XPS analysis) after fluoride adsorption suggested the major adsorption mechanism of ligand exchange. Besides, the adsorption/desorption cycle studies demonstrated the well-retained performance of Mag@Al2Fe for repeated application after regeneration by 0.5 mol/L NaOH solution. Facile synthesis, high defluoridation, lower cost, and quick separation of Mag@Al2Fe indicates its promising potential for drinking water defluoridation.

6.
Int J Oncol ; 54(3): 1086-1098, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30628664

RESUMO

Globally, gastric cancer is the fifth most common malignancy, with high rates of incidence and mortality. The high mortality rate and poor prognosis of gastric cancer are closely associated with its profound invasiveness, high incidence of metastasis, rapid proliferation, and high rate of recurrence. Previous studies have confirmed that stathmin (STMN) has an important role in the occurrence, development and prognosis of gastric cancer. However, the detailed mechanisms by which STMN affects these processes remain unclear. The aim of the present study was to determine how STMN promotes invasion, migration and proliferation in gastric cancer tumor cells. The results of immunohistochemistry indicated that STMN is overexpressed in stomach neoplasm tissues, and that it is associated with migration, invasion, proliferation and anti­apoptotic states of gastric cancer cells. The secretory proteins of gastric cancer cells with or without STMN knockdown were further analyzed using the isobaric tags for relative and absolute quantitation method to identify differentially expressed proteins verified by reverse transcription­quantitative polymerase chain reaction and western blot analysis. Inhibition of STMN decreases the levels of clusterin, cystatin C and matrix metalloproteinases, followed by inhibiting the protein kinase B and signal transducer and activation of transcription activation. These findings suggest that STMN could be a promising therapeutic target for gastric cancer.


Assuntos
Clusterina/metabolismo , Inativação Gênica , Proteínas Proto-Oncogênicas c-akt/metabolismo , Fator de Transcrição STAT3/metabolismo , Transdução de Sinais/genética , Estatmina/genética , Neoplasias Gástricas/patologia , Idoso , Apoptose , Biomarcadores Tumorais/genética , Biomarcadores Tumorais/metabolismo , Movimento Celular/genética , Proliferação de Células/genética , Clusterina/genética , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Invasividade Neoplásica/genética , Proteínas Proto-Oncogênicas c-akt/genética , Fator de Transcrição STAT3/genética , Estatmina/metabolismo , Neoplasias Gástricas/metabolismo
7.
Cell Physiol Biochem ; 48(2): 741-752, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30025407

RESUMO

BACKGROUND/AIMS: C reactive protein (CRP) levels are elevated in many diseases, including malignant tumors and cardiovascular disorders. In this study, the protein interaction network for CRP was evaluated to determine the importance of CRP and its interacting proteins in the molecular pathogenesis of hepatocellular carcinoma (HCC). METHODS: Isobaric tags for relative and absolute quantitation (iTRAQ) and mass spectrometry were used to identify CRP interacting proteins in SMMC7721 cells. Moreover, Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) were used to evaluate enriched genes and pathways for differentially expressed genes using DAVID and WebGestalt. Co-immunoprecipitation and western blot analyses were employed to assess interactions between CRP and KRT8, ANXA2, ENO2, and HSP90B1. RESULTS: In total, 52 proteins that interact with CRP were identified. A GO analysis suggested that most of the interacting proteins were involved in CRP complexes and regulated metabolic processes. A KEGG pathway analysis suggested that most CRP-interacting proteins contribute to the TRAIL signaling pathway, Class I PI3K/Akt signaling pathway, plasma membrane estrogen receptor signaling, Nectin adhesion pathway, and S1P1 pathway. Immunoprecipitation and western blot analyses revealed interactions between CRP and KRT8, ANXA2, ENO2, and HSP90B1. CONCLUSIONS: iTRAQ based proteomic profiling revealed the network of CRP interacting proteins. This network may activate the PI3K/Akt signaling pathway, thereby contributing to the pathogenesis of HCC.


Assuntos
Proteína C-Reativa/metabolismo , Carcinoma Hepatocelular/patologia , Neoplasias Hepáticas/patologia , Proteômica , Anexina A2/metabolismo , Proteína C-Reativa/antagonistas & inibidores , Proteína C-Reativa/genética , Carcinoma Hepatocelular/metabolismo , Perfilação da Expressão Gênica , Humanos , Queratina-8/metabolismo , Neoplasias Hepáticas/metabolismo , Glicoproteínas de Membrana/antagonistas & inibidores , Glicoproteínas de Membrana/genética , Glicoproteínas de Membrana/metabolismo , Nectinas/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Mapas de Interação de Proteínas , Proteínas Proto-Oncogênicas c-akt/metabolismo , Interferência de RNA , RNA Interferente Pequeno/metabolismo , Transdução de Sinais , Ligante Indutor de Apoptose Relacionado a TNF/metabolismo
8.
J Colloid Interface Sci ; 530: 704-713, 2018 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-30015156

RESUMO

Present study reports the successful development of a novel lanthanum (La)-based magnetic adsorbent and its use for phosphate removal from water. For its synthesis, natural magnetite (Mag), Fe3O4, was subjected to partial dissolution in HCl solution and the obtained suspension was mixed with an alkaline solution for in-situ synthesis of ferrihydrite (Fh)-coated Mag (Mag@Fh). Mag@Fh was then decorated with La (hydr)oxides followed by calcination to produce Fh-coated and La-decorated Mag (Mag@Fh-La). Obtained Mag@Fh-La represented high phosphate adsorption capacity (44.8 mg P/g at 15.7% La in its structure) and La usage efficiency. Moreover, Mag@Fh-La retained its high adsorption capacity (>35.0 mg P/g) over a wide range of equilibrium solution pH (3.2-10.7). The combination of FTIR, XPS analysis and adsorption experiments revealed that ligand exchange and electrostatic attraction were the main mechanisms that jointly facilitated the adsorption of phosphate. Adsorption-desorption cycle studies confirmed the well-retained adsorption efficiency of regenerated Mag@Fh-La for repeated applications. Final experiments with real domestic wastewater (initial phosphate concentration of 1.7 mg/L) revealed that 0.2 g/L Mag@Fh-La efficiently reduced the phosphate concentration to below 0.02 mg/L. Overall, this work clearly highlights that the synthesized novel adsorbent has promising applications in phosphate removal from real wastewater.

9.
Int J Oncol ; 53(1): 266-274, 2018 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-29749468

RESUMO

Metastasis is a characteristic of malignant tumors and may be a fatal clinical factor for many patients with cancer. Hepatocellular carcinoma (HCC) cells are highly metastatic; the mechanism of metastasis is complicated and may be influenced by a number of factors. Membrane proteins may block receptors or inhibit important enzymes, thus inhibiting tumor progression, and may be potential therapeutic targets for tumor prognosis and treatment. The present study aimed to use proteomics to analyze the dynamic changes of membrane proteins in HCC cells, to improve our understanding of membrane protein functions and to clarify the important components of the mechanisms of HCC metastasis. The present study used the highly metastatic MHCC97-H and the lowly metastatic MHCC97-L HCC cell lines, and the isobaric tags for relative and absolute quantitation (iTRAQ) approach was used for high-throughput screening of metastasis-related membrane proteins. A total of 22 membrane proteins were identified as differentially expressed between the MHCC97-H and MHCC97-L cell lines; these results were verified by reverse transcription-quantitative polymerase chain reaction and western blotting. A number of the identified proteins were revealed to be related to tumor metastasis, including the tetraspan in transmembrane protein CD9. CD9 was demonstrated to be highly expressed in MHCC97-H cells compared with MHCC97-L cells. The functional role of CD9 was characterized by inhibiting its expression using a small interfering RNAs, which demonstrated that reduced CD9 expression inhibited cell migration and metastasis, as determined by wound-healing and invasion assays. Results from the present study demonstrated that CD9 was highly expressed in the highly metastatic HCC cells and promoted HCC cell migration. This protein may be a novel target for regulating the invasive phenotype in HCC.


Assuntos
Carcinoma Hepatocelular/genética , Neoplasias Hepáticas/genética , Proteínas de Membrana/genética , Tetraspanina 29/genética , Carcinoma Hepatocelular/patologia , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células/genética , Regulação Neoplásica da Expressão Gênica , Ensaios de Triagem em Larga Escala , Humanos , Neoplasias Hepáticas/patologia , Invasividade Neoplásica/genética , Metástase Neoplásica , RNA Interferente Pequeno/genética , Tetraspanina 29/antagonistas & inibidores
10.
Cell Physiol Biochem ; 45(2): 744-760, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29414802

RESUMO

BACKGROUND/AIMS: Hepatitis B virus (HBV) infection is a major cause of cirrhosis and hepatocellular carcinoma. Therefore, we aimed to obtain further information on HBV pathogenesis, and to search for novel putative molecules for anti-HBV therapy. METHODS: We utilized Isobaric Tags for Relative and Absolute Quantitation (iTRAQ) to identify the secretory proteins that are differentially expressed in the HBV DNA-transfected HepG2.2.15 cell line and its parental HepG2 cell line. Immunohistochemistry (IHC) was employed to assess the clinical relevance of the observations. Small interfering (si)RNA-based silencing transfection methods were carried out to study the function of ENPP2. RESULTS: Totally, 133 unique proteins were identified as differentially expressed in HepG2.2.15 cell line compared with HepG2 cell line. Ectonucleotide pyrophosphatase/phosphodiesterase family member 2 precursor (ENPP2) is one of the most significantly up-regulated secretory proteins associated with HBV replication. This differential expression of ENPP2 was further validated by real-time quantitative RT-PCR, Western Blot and immunohistochemical analysis. To study the function of ENPP2, we knockdown ENPP2 expression in HepG2.2.15 cell line by RNA interference. ENPP2 silencing increased HBV replication approximately 2.3-fold by enhancing, via the type I IFN signaling pathway, HBV cccDNA (covalently closed circular DNA) translation into viral RNA. Moreover, attenuation of ENPP2 expression inhibited both the invasion and migration ability of hepatoma cells in vitro via interacting with the molecules in the tumor microenvironment. CONCLUSION: Our study demonstrates that ENPP2 may be a novel anti-HBV target and indicate that suppression of its expression may inhibit the invasion and migration ability of hepatoma cells.


Assuntos
Carcinoma Hepatocelular/patologia , Vírus da Hepatite B/fisiologia , Neoplasias Hepáticas/patologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Carcinoma Hepatocelular/sangue , Carcinoma Hepatocelular/metabolismo , Linhagem Celular , Movimento Celular , DNA Viral/fisiologia , Regulação da Expressão Gênica , Células Hep G2 , Humanos , Interferon Tipo I/metabolismo , Neoplasias Hepáticas/sangue , Neoplasias Hepáticas/metabolismo , Proteínas de Membrana/metabolismo , Pessoa de Meia-Idade , Diester Fosfórico Hidrolases/sangue , Diester Fosfórico Hidrolases/química , Diester Fosfórico Hidrolases/genética , Diester Fosfórico Hidrolases/metabolismo , Proteoglicanas/metabolismo , Interferência de RNA , RNA Interferente Pequeno/metabolismo , Transdução de Sinais , Replicação Viral
11.
J Environ Sci (China) ; 63: 1-8, 2018 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-29406093

RESUMO

We report that green algae in lakes and rivers can serve as precursors of halobenzoquinone (HBQ) disinfection byproducts (DBPs) produced during chlorination. Chlorination of a common green alga, Chlorella vulgaris, produced 2,6-dichloro-1,4-benzoquinone (2,6-DCBQ), the most prevalent HBQ DBP in disinfected water. Under varying pH conditions (pH6.0-9.0), 2,6-DCBQ formation ranged from 0.3 to 2.1µg/mg C with maximum formation at pH8.0. To evaluate the contribution of organic components of C. vulgaris to 2,6-DCBQ formation, we separated the organics into two fractions, the protein-rich fraction of intracellular organic matter (IOM) and the polysaccharide-laden fraction of extracellular organic matter (EOM). Chlorination of IOM and EOM produced 1.4µg/mg C and 0.7µg/mg C of 2,6-DCBQ, respectively. The IOM generated a two-fold higher 2,6-DCBQ formation potential than the EOM fraction, suggesting that proteins are potent 2,6-DCBQ precursors. This was confirmed by the chlorination of proteins extracted from C. vulgaris: the amount of 2,6-DCBQ produced is linearly correlated with the concentration of total algal protein (R2=0.98). These results support that proteins are the primary precursors of 2,6-DCBQ in algae, and control of green algal bloom outbreaks in source waters is important for management of HBQ DBPs.


Assuntos
Benzoquinonas/metabolismo , Chlorella vulgaris/fisiologia , Desinfetantes/metabolismo , Poluentes Químicos da Água/metabolismo , Benzoquinonas/análise , Clorófitas , Desinfetantes/análise , Desinfecção , Halogenação , Poluentes Químicos da Água/análise , Purificação da Água/métodos
12.
Mol Med Rep ; 17(3): 3481-3488, 2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-29286136

RESUMO

Despite the use of adjuvant therapies, the cumulative proportion of live births remains at ~40%. Accumulating data show that low pregnancy rates, even in the presence of high fertility rates, are due to implantation failure. The present study aimed to identify and construct a profile of proteins that react with preimplantation factor (PIF) and to provide an understanding into the molecular mechanisms by which PIF promotes trophoblast invasion. Cytoplasmic proteins were immunoprecipitated with biotin­labeled synthetic PIF or intralipid and scrambled PIF (PIFscr). The protein profiles were analyzed using isobaric tags for relative and absolute quantification coupled with mass spectrometry. Immunoprecipitation and western blot analyses were used to assess the interactions between PIF and myosin heavy chain 10 (MYH10) and heat shock protein family D1. Small interfering RNA­based silencing was performed to examine the function of MYH10. In the results of the present study, 21 proteins were identified with interactions with PIF. The immunoprecipitation and western blot analyses revealed an interaction between PIF and MYH10. Silencing of the expression of MYH10 in HEC­1­B cells significantly attenuated cell migration and invasion capacities. These data support the conclusion that MYH10­mediated cell migration and invasion act in conjunction with PIF to promote the trophoblast invasion procedure.


Assuntos
Implantação do Embrião/efeitos dos fármacos , Peptídeos/farmacologia , Proteoma/metabolismo , Linhagem Celular , Movimento Celular/efeitos dos fármacos , Cromatografia Líquida de Alta Pressão , Embrião de Mamíferos/citologia , Embrião de Mamíferos/efeitos dos fármacos , Embrião de Mamíferos/metabolismo , Feminino , Células HEK293 , Humanos , Espectrometria de Massas , Cadeias Pesadas de Miosina/antagonistas & inibidores , Cadeias Pesadas de Miosina/genética , Cadeias Pesadas de Miosina/metabolismo , Miosina não Muscular Tipo IIB/antagonistas & inibidores , Miosina não Muscular Tipo IIB/genética , Miosina não Muscular Tipo IIB/metabolismo , Proteoma/efeitos dos fármacos , Proteômica , Interferência de RNA , RNA Interferente Pequeno/metabolismo , Tubulina (Proteína)/genética , Tubulina (Proteína)/metabolismo
13.
Dig Liver Dis ; 49(7): 780-788, 2017 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-28377286

RESUMO

BACKGROUND: Liver fibrosis can lead to cirrhosis and hepatocellular carcinoma if not treated in the early stages. The molecular mechanisms of the pathogenesis of hepatic fibrosis remain unclear. AIM: To identify the molecules involved in the pathogenesis of liver fibrosis and to investigate the potential effect and mechanism of Annexin A2 up-regulation during liver fibrosis progression. METHODS: Twenty Sprague-Dawley rats were divided into two groups: the carbon tetrachloride (CCl4)-induced liver fibrosis group and the normal control group. Hematoxylin and eosin staining or Masson Trichrome staining and enzyme-linked immunosorbent assay were applied to assess the degree of liver damage and fibrosis in rats with CCl4-induced liver fibrosis. Liver tissue protein profiles were analyzed using iTRAQ and mass spectrometry. RT-PCR and western blotting analyses were employed to validate differentially expressed proteins. Small interfering RNA-based silencing was performed to study the function of Annexin A2. RESULTS: Twelve weeks after CCl4 injection, significant body weight changes and liver injury and liver fibrosis were observed in rats. In addition, 130 proteins were differentially expressed in the liver fibrosis group. Overexpression of Annexin A2 was confirmed by RT-PCR and Western blotting analysis. Silencing of Annexin A2 expression in HepG2 and LX-2 cells significantly reduced the secretion of von Willebrand factor (vWF). CONCLUSION: Annexin A2 promotes liver fibrosis by mediating vWF secretion, which can be used to mitigate the progression of liver fibrosis.


Assuntos
Anexina A2/metabolismo , Cirrose Hepática Experimental/metabolismo , Fator de von Willebrand/metabolismo , Animais , Western Blotting , Modelos Animais de Doenças , Ensaio de Imunoadsorção Enzimática , Inativação Gênica , Cirrose Hepática Experimental/etiologia , Espectrometria de Massas , Ligação Proteica , RNA Interferente Pequeno , Distribuição Aleatória , Ratos , Ratos Sprague-Dawley , Regulação para Cima
14.
Int J Oncol ; 2017 Mar 13.
Artigo em Inglês | MEDLINE | ID: mdl-28350119

RESUMO

Hepatocellular carcinoma (HCC) is one of most common malignant cancers and is the second leading cause of cancer related deaths. The prognosis and survival of patients are closely related to the degree of tumor metastasis. The mechanism of HCC metastasis is still unclear. In the present study, we investigated the molecular mechanism of C-reaction protein in promoting migration and invasion of hepatocellular carcinoma cells in vitro. We estimated that CRP is overexpressed in liver cancer tissues and that it promotes invasion and metastasis of HCC in vitro. In the present study, we employed iTRAQ-based mass spectrometry to analyze the HepG2 secretory proteins of CRP siRNA-treated cells and negative control siRNA-treated cells. We identified 109 differentially expressed proteins after silencing CRP, of which 45 were upregulated and 64 were downregulated. Some of the differentially expressed proteins were confirmed by western blot analysis and real-time quantitative PCR. Furthermore, we found that knockdown of CRP substantially abrogates HIF-1α expression levels, the luciferase activity of HIF-1α and ERK and Akt phosphorylation in HepG2 cells. The present study provides a novel mechanism by which CRP promotes the proliferation, migration, invasion and metastasis of hepatocellular carcinoma cells. Inhibition of CRP suppressed migration, invasion and healing of hepatoma carcinoma cells by decreasing HIF-1α activity and CTSD.

15.
Int J Oncol ; 50(3): 883-892, 2017 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-28197637

RESUMO

Hepatocellular carcinoma is the second most common cause of cancer-related deaths worldwide. Due to a high propensity to metastasize, active angiogenesis and rapid proliferation, recurrence and poor prognosis are major obstacles for treatment and cure of this disease. However, the detailed mechanisms of how fatty acid synthase (FASN) promotes migration, invasion and healing in tumor cells remain unclear. In the present study, the previous results that FASN was expressed higher in cancer samples than in non-cancerous samples, and influenced migration, invasion of hepatoma carcinoma cells, were verified by immunohistochemistry, tissue microarrays, Transwell assay and wound healing assay. The secretory proteins of hepatocellular carcinoma cells with or without FASN knockdown were analyzed using the isobaric tags for relative and absolute quantitation (iTRAQ) method to identify differentially expressed proteins (DEPs). The DEPs were verified by RT-PCR and western blot analysis, and were consistent with the iTRAQ results. Inhibition of FASN can decrease the levels of IGFBP1, and the expression, activity, and ubiquitination of HIF-1α. Inhibition of FASN can suppress migration, invasion and healing of hepatoma carcinoma cells by decreasing HIF-1α, and IGFBP1.


Assuntos
Carcinoma Hepatocelular/patologia , Movimento Celular/genética , Ácido Graxo Sintase Tipo I/antagonistas & inibidores , Ácido Graxo Sintase Tipo I/genética , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Proteína 1 de Ligação a Fator de Crescimento Semelhante à Insulina/metabolismo , Neoplasias Hepáticas/patologia , Carcinoma Hepatocelular/genética , Linhagem Celular Tumoral , Proliferação de Células/genética , Células Hep G2 , Humanos , Neoplasias Hepáticas/genética , Invasividade Neoplásica/genética , Interferência de RNA , RNA Interferente Pequeno/genética
16.
PLoS One ; 12(2): e0172214, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28222113

RESUMO

Cortical dysplasia accounts for at least 14% of epilepsy cases, and is mostly seen in children. However, the understanding of molecular mechanisms and pathogenesis underlying cortical dysplasia is limited. The aim of this cross-sectional study is to identify potential key molecules in the mechanisms of cortical dysplasia by screening the proteins expressed in brain tissues of childhood cortical dysplasia patients with epilepsy using isobaric tags for relative and absolute quantitation-based tandem mass spectrometry compared to controls, and several differentially expressed proteins that are not reported to be associated with cortical dysplasia previously were selected for validation using real-time polymerase chain reaction, immunoblotting and immunohistochemistry. 153 out of 3340 proteins were identified differentially expressed between childhood cortical dysplasia patients and controls. And FSCN1, CRMP1, NDRG1, DPYSL5, MAP4, and FABP3 were selected for validation and identified to be increased in childhood cortical dysplasia patients, while PRDX6 and PSAP were identified decreased. This is the first report on differentially expressed proteins in childhood cortical dysplasia. We identified differential expression of FSCN1, CRMP1, NDRG1, DPYSL5, MAP4, FABP3, PRDX6 and PSAP in childhood cortical dysplasia patients, these proteins are involved in various processes and have various function. These results may provide new directions or targets for the research of childhood cortical dysplasia, and may be helpful in revealing molecular mechanisms and pathogenesis and/or pathophysiology of childhood cortical dysplasia if further investigated.


Assuntos
Química Encefálica , Epilepsias Parciais/metabolismo , Perfilação da Expressão Gênica , Malformações do Desenvolvimento Cortical/metabolismo , Proteínas do Tecido Nervoso/biossíntese , Proteômica/métodos , Western Blotting , Criança , Pré-Escolar , Anormalidades Craniofaciais , Estudos Transversais , Epilepsias Parciais/genética , Feminino , Ontologia Genética , Humanos , Técnicas Imunoenzimáticas , Masculino , Malformações do Desenvolvimento Cortical/genética , Proteínas do Tecido Nervoso/genética , Proteínas do Tecido Nervoso/isolamento & purificação , Reação em Cadeia da Polimerase em Tempo Real , Espectrometria de Massas em Tandem
17.
Oncotarget ; 8(3): 4549-4562, 2017 Jan 17.
Artigo em Inglês | MEDLINE | ID: mdl-27999186

RESUMO

BACKGROUND AND AIMS: Hepatitis B virus (HBV) infection is a major risk factor for liver cirrhosis and hepatocellular carcinoma (HCC). To gain a better understanding of the pathogenesis of HBV infection, this study aimed to investigate the differentially expressed proteins (DEPs) in liver tissues from patients with chronic hepatitis B (CHB) infection. RESULTS: Seventy-one DEPs were identified. Overexpression of multi-drug resistance protein 1 (MDR1) was validated by RT-qPCR and western blot analyses. Moreover, its expression was increased at both the mRNA and protein levels in response to overexpression of HBV large surface protein (LHBs). Furthermore, screening of transcription factors suggested the possible involvement of hypoxia-inducible factor 1α (HIF-1α) in the interaction between LHBs and MDR1. The function of HIF-1α in the MDR1 activation was confirmed by EMSA and reporter gene analyses. MATERIALS AND METHODS: Liver samples from CHB patients and controls without HBV infection were collected and subjected to isobaric tags for relative and absolute quantitation (iTRAQ) and mass spectrometric analysis. CONCLUSIONS: These results imply that LHBs, in association with HIF-1α, induces MDR1 overexpression, which may contribute to the pathogenic changes in CHB infection.


Assuntos
Carcinoma Hepatocelular/virologia , Hepatite B Crônica/metabolismo , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Neoplasias Hepáticas/virologia , Proteínas do Envelope Viral/metabolismo , Subfamília B de Transportador de Cassetes de Ligação de ATP/genética , Subfamília B de Transportador de Cassetes de Ligação de ATP/metabolismo , Adolescente , Adulto , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/metabolismo , Estudos de Casos e Controles , Feminino , Regulação Neoplásica da Expressão Gênica , Células Hep G2 , Vírus da Hepatite B/metabolismo , Vírus da Hepatite B/patogenicidade , Hepatite B Crônica/genética , Humanos , Fígado/metabolismo , Fígado/virologia , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/metabolismo , Masculino , Espectrometria de Massas , Pessoa de Meia-Idade , Proteômica/métodos , Adulto Jovem
18.
Water Sci Technol ; 74(11): 2727-2735, 2016 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-27973377

RESUMO

To access better removal of nutrients with algae-based techniques, a dominant alga from real municipal wastewater was identified and its capacity in removing low concentrations of nitrogen (NH+4 or NO-3) and phosphorus (PO3-4) was evaluated. Results showed that Oedogonium brevicingulatum, a filamentous green alga, was confirmed as the dominant alga in the secondary effluent of a municipal wastewater treatment plant by polymerase chain reaction-denaturing gradient gel electrophoresis. Low concentrations of NH+4 or NO-3 (≤5 mg N L-1) and PO3-4 (≤0.5 mg P L-1) were 100% removed by the algae in a 7-d test. The maximum nutrient removal rate (Vmax) and the half-saturation constant (Km) for NH+4 (10.03 ± 0.95 mg g-1d-1 and 0.19 ± 0.03 mg L-1) and NO-3 (8.43 ± 0.21 mg g-1 d-1 and 0.27 ± 0.11 mg L-1) indicated the uptake capability for NH+4 is higher than that for NO-3. Meanwhile, it showed higher affinity for PO3-4 (Vmax: 1.42 ± 0.02 mg g-1 d-1; Km: 0.02 ± 0.00 mg L-1) with NH+4 as nitrogen source than that (Vmax: 1.24 ± 0.15 mg g-1 d-1; Km: 0.06 ± 0.03 mg L-1) with NO-3 as nitrogen source. Moreover, nutrient removal efficiencies were observed steady when nitrogen/phosphorus ratio ranged from 5:1 to 20:1. These results suggest that the dominant algae from municipal wastewater have potentials to be applied in nutrient removal.


Assuntos
Clorófitas/metabolismo , Nitrogênio/metabolismo , Fósforo/metabolismo , Águas Residuárias/análise , Poluentes da Água/análise , Poluentes da Água/metabolismo , Compostos de Amônio/metabolismo , Nitratos/metabolismo , Fosfatos/metabolismo , Eliminação de Resíduos Líquidos/métodos
19.
Oncotarget ; 7(46): 75468-75481, 2016 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-27690342

RESUMO

Cervical cancer is one of the most common malignant tumor in women. The mechanisms of cervical cancer are intricate and have not been fully understood. Therefore, we employed iTRAQ to obtain novel proteins profile which participates in the tumor oncogenesis of cervical cancer. 3300 proteins were identified aberrantly expressed in cervical cancer, and western bolt was performed to validate the results of iTRAQ. Then, we selected LYN for further study. Immunohistochemistry identified that LYN expression was significantly increased in cervical cancer tissues than that in cancer adjacent normal cervical tissues and normal cervical tissues. The increased LYN expression was significantly correlated with cancer differentiation and FIGO stage. Silencing LYN inhibited cell proliferation, migration and invasion, conversely, overexpression LYN promoted cell proliferation, migration and invasion. In terms of mechanism, LYN could also promote cervical cancer cells metastasis through activating IL-6/STAT3 pathway. In vivo study, overexpression LYN promoted tumor growth, meanwhile knockdown LYN inhibited tumor growth. These results indicate that LYN tyrosine kinase is an oncogenic gene and can serve as a novel target for cervical cancer research and therapy.


Assuntos
Proteínas Oncogênicas/metabolismo , Neoplasias do Colo do Útero/metabolismo , Quinases da Família src/metabolismo , Antineoplásicos/farmacologia , Antineoplásicos/uso terapêutico , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células , Feminino , Humanos , Interleucina-6/metabolismo , Terapia de Alvo Molecular , Estadiamento de Neoplasias , Proteínas Oncogênicas/antagonistas & inibidores , Proteínas Oncogênicas/genética , Ligação Proteica , Inibidores de Proteínas Quinases/farmacologia , Inibidores de Proteínas Quinases/uso terapêutico , Proteoma , Proteômica/métodos , Fator de Transcrição STAT3/metabolismo , Transdução de Sinais/efeitos dos fármacos , Neoplasias do Colo do Útero/genética , Neoplasias do Colo do Útero/patologia , Quinases da Família src/antagonistas & inibidores , Quinases da Família src/genética
20.
Bioresour Technol ; 220: 246-252, 2016 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-27584901

RESUMO

This study investigated effects of pH-depended inorganic carbon (IC) species and pH on algal growth in the sewage simulation system, and fruitfully discussed the relationships among IC, pH and algal growth by the Monod kinetics. Results showed HCO3(-) significantly increased algal growth by 3.17-6.52 times than that of CO3(2-) and/or glucose when the value of pH was in the range of 8.0-9.5, and also the preferentially utilized indicated by the affinity coefficient (Kp) of HCO3(-), CO3(2-) and glucose (0.17, 15.14 and 31.22, respectively). Meanwhile, the same pH range facilitated HCO3(-) to become a dominated species (e.g., 48.80-93.19% of total IC). More importantly, good linear correlations pairwise existed among pH, IC species and algae growth. These results suggested pH plays a critical role in regulation of IC species and algae growth, which would be an efficient method to control the IC discharge from sewage effluents and weaken bloom outbreak.


Assuntos
Carbono/metabolismo , Chlorella/crescimento & desenvolvimento , Chlorella/metabolismo , Eutrofização , Concentração de Íons de Hidrogênio , Cinética , Modelos Teóricos , Esgotos
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