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1.
Artigo em Inglês | MEDLINE | ID: mdl-31953219

RESUMO

This study aimed to explore how Wuchang bream (Megalobrama amblycephala) survive and defend against the toxicity of ambient total ammonia nitrogen (0, 5, 10, 20 and 30 mg/L TA-N) during 30-day exposure. As a result, hepatic malondialdehyde and protein carbonylation as well as histopathological alterations increased with increasing TA-N level, which suggested that chronic ammonia exposure caused oxidative stress and damage in the liver of fish. Meanwhile, the activities of hepatic total superoxide dismutase (T-SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR) and glucose 6-phosphate dehydrogenase (G6PD) as well as the mRNA expression of Cu/Zn sod, cat, gpx and g6pd were elevated significantly along with significant reduction of glutathione (GSH) and nicotinamide adenine dinucleotide phosphate (NADPH) (P < 0.05). These results indicated that hepatic antioxidant responses were activated to alleviate oxidative damages induced by ammonia, in which lower-concentration ammonia only initiate SOD-CAT-GR-G6PDH defense and higher ammonia activated the SOD-CAT-GPx-GSH-GR-G6PDH antioxidant response. In addition, significant increases of serum urea and hepatic ammonia, urea, glutamine, arginase as well as glutamine synthetase were detected with the increase of TA-N (P < 0.05), while serum ammonia levels kept stable (P > 0.05). The present findings further revealed that ammonia could be detoxified directly into glutamine and urea in Wuchang bream to cope with ammonia exposure. In conclusion, under chronic ammonia exposure, enhanced hepatic antioxidant responses as well as increased urea and glutamine synthesis worked in combination to allow Megalobrama amblycephala to defend against environmental ammonia toxicity.


Assuntos
Amônia , Cyprinidae/metabolismo , Proteínas de Peixes/metabolismo , Glutamina/metabolismo , Fígado , Ureia/metabolismo , Amônia/metabolismo , Amônia/toxicidade , Animais , Fígado/metabolismo , Fígado/patologia , Estresse Oxidativo , Poluentes Químicos da Água/toxicidade
2.
Clin Chim Acta ; 500: 59-68, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31655056

RESUMO

BACKGROUND: Type 2 diabetes (T2D), a typical metabolic disease, is closely associated with serum free fatty acids. But the association between serum total fatty acids (TFAs, free fatty acids plus esterified fatty acids) and T2D has not been reported. METHODS: Serum esterified fatty acids were hydrolyzed under alkaline conditions, and serum TFAs were extracted after acidizing. Fourteen of serum TFAs in 1,828 serum samples, including 543 controls, 655 prediabetes, and 630 T2D patients, were simultaneously quantified based on the calibration curves of 8 fatty acids using matrix-assisted laser desorption/ionization-Fourier transform ion cyclotron resonance mass spectrometry (MALDI-FTICR MS). RESULTS: Correlation analysis revealed strong correlations among serum TFAs and ratios of the TFAs in T2D patients compared with controls or prediabetes both in males and females. Receiver operating characteristic analysis indicated that a panel including fasting plasma glucose, glycosylated hemoglobin type A1c, gamma-glutamyltransferase, triglyceride, C18:1, and C20:3, has a good capability to distinguish prediabetes from T2D, with the sensitivity of 87.0%, the specificity of 91.0%, and the area under curve (AUC) of 0.96. CONCLUSIONS: In this study, rapid, absolute, and simultaneous quantification of serum TFAs was performed using MALDI-FTICR MS. C18:1 and C20:3 were significantly correlated with prediabetes and T2D.

3.
Sci Total Environ ; 702: 134969, 2020 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-31710851

RESUMO

Waterborne microcystin-LR (MC-LR) released by cyanobacterial blooms in eutrophic water bodies have caused serious risk to aquatic animal and human health. In the present study, we for the first time conducted a comprehensive in vivo investigation on chronic inflammatory responses and its molecular pathways of different environmental relevant levels of MC-LR (0, 0.4, 2 and 10 µg/L) in male zebrafish (Danio rerio). The results showed that chronic MC-LR exposure caused splenic inflammatory changes including the formation of melano-macrophage centers, remarkable elevation of serum tumor necrosis factor alpha (TNFα) and interleukin 1 beta (IL1ß) levels as well as significant upregulated expression of MyD88-dependent toll-like receptor (TLR/MyD88) signaling pathway genes (tlr4a, myd88, erk2, p38a, il1ß and tnfα). The immunohistochemical and western blot results further validated that higher MC-LR concentrations tended to enhance the MyD88 signal. Moreover, significant decreases of serum C3 levels along with splenic c3b expression in the 10 µg/L exposure group proved that chronic MC-LR exposure could ultimately decrease the innate immunity of fish. Our findings revealed that chronic exposure of MC-LR could cause chronic inflammation through TLR/MyD88 signaling pathway and subsequently induce immune disorders in male zebrafish, which also urge us to pay more attention on the potential immunotoxicity of long-term exposure to low concentration of MC-LR.


Assuntos
Microcistinas/toxicidade , Fator 88 de Diferenciação Mieloide/metabolismo , Poluentes Químicos da Água/toxicidade , Proteínas de Peixe-Zebra/metabolismo , Animais , Masculino , Peixe-Zebra
4.
Environ Sci Technol ; 54(2): 1014-1023, 2020 01 21.
Artigo em Inglês | MEDLINE | ID: mdl-31859493

RESUMO

Transgenerational effects of microcystin-LR (MC-LR) released by cyanobacterial blooms have become a hot topic. In the present study, adult zebrafish pairs were exposed to 0, 0.4, 2, and 10 µg/L MC-LR for 60 days and the embryos (F1 generation) were hatched without or with continued MC-LR exposures at the same concentrations until 5 days postfertilization (dpf). The results showed the existence of MC-LR both in F0 gonads and in F1 embryos and indicated that MC-LR could be transferred directly from the F0 adult fish to F1 offspring. The adverse effects on sex hormone levels, sexual development, and fecundity in F0 generation along with abnormal development in F1 offspring were observed. Furthermore, downregulation of antioxidant genes (cat, mn-sod, gpx1a) and upregulation of innate immune-related genes (tlr4a, myd88, tnfα, il1ß) as well as increased proinflammation cytokine contents (TNF-α, IL-1ß, IL-6) were noticed in F1 offspring without/with continued MC-LR exposures. In addition, significant differences between the two F1 embryo treatments demonstrated that continuous MC-LR exposure could result in a higher degree of inflammatory response compared to those without MC-LR exposure. Our findings revealed that MC-LR could exert cross-generational effects of immunotoxicity by inhibiting the antioxidant system and activating an inflammatory response.


Assuntos
Poluentes Químicos da Água , Peixe-Zebra , Animais , Imunidade Inata , Microcistinas , Oxirredutases
5.
Clin Chim Acta ; 489: 124-129, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30543790

RESUMO

BACKGROUND: The goal of this study was to detect novel modified forms of hemoglobin using mass spectrometry (MS) and to investigate the effect of modified hemoglobin on HbA1c and fasting plasma glucose (FPG). METHODS: This study was conducted on 1200 subjects aged >25 years. Hemoglobin from the above-mentioned subjects was detected using direct-infusion electrospray ionization-MS, and HbA1c and FPG were measured according to the manufacturer's instructions. Regression analysis was performed to estimate the correlations and interactions among HbA1c, FPG, and modified hemoglobin. RESULTS: Multiple modified forms (α1, α2, α3, ß1, ß2, and ß3) of hemoglobin were observed using MS. Statistical analyses indicated that modified hemoglobin was significantly correlated with FPG (p ≤ .01). The association of FPG with α1% (p = .021) and ß3% (p < .001) values was independent of HbA1c% and other modified forms of hemoglobin. Interaction analyses implied two significant interaction effects of HbA1c% with gender (ß = -0.184, p = .007) and α3% (ß = -0.104, p < .001) on FPG. The relationship between HbA1c% and FPG was stronger in males than in females, and a decreased level of α3% also affected the association of HbA1c% and FPG. CONCLUSIONS: This MS-based method is an effective tool for monitoring glycated forms of hemoglobin than traditional approaches. For the Han Chinese population, multiple-glycated hemoglobin affects the association of FPG with HbA1c%, and the correlation between FPG and HbA1c% in females is different from that in males. These data suggest that the HbA1c criteria for the diagnosis and monitoring of diabetes should be established according to genders and modified types of hemoglobin.


Assuntos
Análise Química do Sangue/métodos , Glicemia/análise , Grupos Étnicos , Hemoglobinas/análise , Espectrometria de Massas , Adulto , China/etnologia , Jejum/sangue , Feminino , Hemoglobina A Glicada/análise , Humanos , Masculino , Pessoa de Meia-Idade
6.
Chemosphere ; 183: 315-322, 2017 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-28551208

RESUMO

Microcystins (MCs), produced by toxic cyanobacterial blooms that appeared world wildly in eutrophication waters, have often caused fish illness and even massive death cases. Among at least 90 structural variants, microcystin-LR (MC-LR) is the most common and toxic variant. In order to better understand innate immune responses in fish disrupted by environmental concentrations of MC-LR, male zebrafish (Danio rerio) were exposed to 0, 0.3, 1, 3, 10 and 30 µg/L MC-LR for 30 d, and the changes in splenic pathology and immunological gene expression as well as serum immune parameters were studied. In the low concentration groups (0.3, 1 and 3 µg/L), zebrafish displayed splenic inflammatory changes including the formation of melano-macrophage centers and the increase of macrophage pseudopodia, remarkable elevation of serum C3 levels, and significantly upregulated expression of innate immune-related genes (c3b, lyz, il1ß, tnfα and ifnγ). In contrast, high concentrations of MC-LR (10 and 30 µg/L) resulted in the degeneration of splenic lymphocytes and macrophages, and down-regulation of immune-related genes as well as significant decreases in the level of serum C3. Furthermore, significant increases in the activity of serum ACP and ALP suggested that high concentrations of MC-LR increased permeability of macrophage plasma membrane or cellular necrosis, and subsequently decreased innate immune function. Our findings illustrated that sub-chronic exposure of MC-LR has dualistic influences on fish innate immune system with inflammatory activation at low exposure concentrations but turned to immune inhibition with the increases of exposure concentration.


Assuntos
Peixes/microbiologia , Imunidade Inata/efeitos dos fármacos , Imunomodulação , Microcistinas/farmacologia , Peixe-Zebra/imunologia , Animais , Exposição Ambiental/efeitos adversos , Peixes/imunologia , Regulação da Expressão Gênica/imunologia , Imunidade Inata/genética , Masculino , Modelos Animais , Baço/patologia , Ativação Transcricional/imunologia , Peixe-Zebra/metabolismo
7.
Zhonghua Wai Ke Za Zhi ; 45(16): 1128-31, 2007 Aug 15.
Artigo em Chinês | MEDLINE | ID: mdl-18005619

RESUMO

OBJECTIVE: To observe the mechanical properties of decellularized porcine aortic valve, and to explore the effects of precoating methods of biological scaffold on histocompatibility. METHODS: Fresh porcine aortic valves were decellularized using trypsin, TritonX-100 and nuclease. Treated valves were evaluated by light microscopy, scanning electron microscopy (SEM) and mechanical test. Three groups of scaffold were precoated with phosphate buffered saline (PBS), poly-L-lysine (PLL) and fetal bovine serum (FBS) respectively. Myofibroblasts were seeded onto each scaffold. Light and electron microscopic observation was performed and MTT test was used to examine efficiency of cell attachment. RESULTS: HE stain and SEM showed that cells were almost absent in the treated leaflet. The wave-like collagen together with the whole three-dimensional structure was maintained. Compared with normal valves, the Max-load, Max-stress and elastic modulus decreased while the Max-strain increased (P<0.05). The result of MTT test showed more cells were attached on the valves treated with FBS compared to the other two groups. Histological investigations also confirm that the high degree of cell attachment on the valves precoated with FBS (F=129.26, P=0.000). CONCLUSIONS: Enzyme combined with detergent and nucleases can remove cells from porcine aortic valves. Meanwhile the mechanical properties of these valves may be altered. Precoating porcine aortic valve with FBS is an effective method to improve cell attachment, growth and increasing.


Assuntos
Valva Aórtica/fisiologia , Materiais Revestidos Biocompatíveis/química , Tecidos Suporte/química , Animais , Valva Aórtica/citologia , Valva Aórtica/efeitos dos fármacos , Fenômenos Biomecânicos , Bioprótese , Adesão Celular , Proliferação de Células , Células Cultivadas , Materiais Revestidos Biocompatíveis/farmacologia , Fibroblastos/citologia , Próteses Valvulares Cardíacas , Ratos , Suínos , Engenharia Tecidual/métodos
8.
Zhonghua Jie He He Hu Xi Za Zhi ; 30(7): 518-21, 2007 Jul.
Artigo em Chinês | MEDLINE | ID: mdl-17961406

RESUMO

OBJECTIVE: To analyze the association of the polymorphism of Met764Thr with bronchial asthma and lung function of asthmatic subjects of Han nationality in Southern China. METHODS: In 164 unrelated patients with asthma and 112 unrelated healthy controls, polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and DNA sequencing were used to determine polymorphism of Met764Thr locus allele in ADAM33 gene. The clinical indexes associated with lung function (FVC%, FEV(1)%) were compared among the three genotypes (Met764/Met764, Met764/Thr764, Thr764/Thr764) in asthmatic subjects. RESULTS: No significant difference was found in the allele (Met764, Thr 764) frequency among populations in UK, US, German, Korean, and Southern China (chi(2) = 6.77, P > 0.05). The frequencies of the genotypes (Met764/Met764, Met764/Thr764, Thr764/Thr764) were respectively 78.7% (129), 18.3% (30), 3.0% (5) in 164 asthmatic subjects and respectively 91.1% (102), 6.3% (7), 2.7% (3) in the 112 controls. There was a significant difference in the distributions of the genotypes (Met764/Met764, Met764/Thr764, Thr764/Thr764) between asthmatic subjects and controls (chi(2) = 8.46, P < 0.05). The frequencies of alleles (Thr764) were respectively 12.2% in the asthmatic subjects and 5.8% in the controls. Significant difference was observed in the allele (Met764, Thr 764) frequency between the two groups (chi(2) = 6.27, P < 0.05). The presence of Thr764 allele of ADAM33 gene was found to be a greater risk factor in asthmatic subjects than in controls. The odds ratio (OR) of Met764/Thr764 and Met764/Thr764 + Thr764/Thr764 were 3.389 (1.430 - 8.030), 2.767 (1.308 - 5.854), respectively. When compared with Met764/Met764 genotype, all P < 0.05. There was a significant decrease in the FVC% and FEV(1)% levels of Met764/Thr764, Thr764/Thr764 and Met764/Met764 genotype. CONCLUSIONS: These results suggest that Met764Thr locus genetic polymorphism is associated with susceptibility of asthma and clinical indexes of lung function of asthmatic subjects of Han nationality in Southern China.


Assuntos
Proteínas ADAM/genética , Asma/genética , Asma/fisiopatologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Grupo com Ancestrais do Continente Asiático , Estudos de Casos e Controles , Feminino , Frequência do Gene , Predisposição Genética para Doença , Genótipo , Humanos , Masculino , Pessoa de Meia-Idade , Polimorfismo de Fragmento de Restrição , Testes de Função Respiratória , Adulto Jovem
9.
Zhonghua Yi Xue Za Zhi ; 87(23): 1622-6, 2007 Jun 19.
Artigo em Chinês | MEDLINE | ID: mdl-17803853

RESUMO

OBJECTIVE: To investigate the effects of transforming growth factor-beta1 (TGF-beta1) on construction of tissue engineering heart valves (TEHV). METHODS: Fresh porcine aortic valves were decellularized with trypsinase and detergent Triton X-100. Myofibroblasts were obtained from rat thoracic aorta, cultured, transfected with the vector containing TGF-beta1 gene-plasmid pcDNA3.0/TGF-beta1 mediated by lipofectamine 2,000 after 48 hours, screened by G418 for for 3 weeks. Decellularized valves were divided into 3 groups: Group A, seeded with the transfected myofibroblasts and cultured in medium without TGF-beta1, Group B, seeded with the transfected myofibroblasts and cultured in medium with TGF-beta1 10 ng/ml, and Group C, seeded with non-transfected myofibroblasts and cultured in medium without TGF-beta1. Hematoxylin-eosin staining and transmission electron microscopy were performed to observe the cell proliferation. DNA contents were measured. Hydroxyproline content was measured so as to indirectly test the collagen production. AGS-J mechanical testing instrument was used to test the mechanical properties of the strips of valves. RESULTS: Immunohistological investigation showed instant TGF-beta1 expression in the myofibroblasts 48 h after the transfection and stable TGF-beta1 expression 4 weeks later. Morphological examination showed that the myofibroblasts in Groups A and B were connected to one another closely with abundant extracellular matrix in the valves. The DNA contents of Groups A and B were (0.126 +/- 0.013) per thousand, and (0.109 +/- 0.004) per thousand, both significantly higher than that of Group [(0.089 +/- 0.011) per thousand, both P < 0.011], with a significant difference between Groups A and B (P < 0.05). The hydroxyproline content of Groups A and B were (5.83 +/- 0.67) per thousand and (5.02 +/- 0.40) per thousand, both significantly higher than that of Group C [(4.34 +/- 0.47) per thousand, both P < 0.05], with a significant difference between Groups A and B (P < 0.05). The maximum load of Groups A and B were (13.4 +/- 1.0) N and (11.7 +/- 1.4) N respectively, both significantly higher than that of Group C [(10.0 +/- 1.1) N, both P < 0.05], with a significant difference between Groups A and B (P < 0.05). CONCLUSION: TGF-beta1 is an important and effective bioactive factor for cell proliferation and extracellular matrix growth of heart valve. It is of great value for constructing TEHV in vitro.


Assuntos
Próteses Valvulares Cardíacas , Engenharia Tecidual/métodos , Fator de Crescimento Transformador beta1/fisiologia , Animais , Células Cultivadas , Fibroblastos/citologia , Fibroblastos/metabolismo , Masculino , Ratos , Ratos Sprague-Dawley , Transfecção , Fator de Crescimento Transformador beta1/genética
10.
Nan Fang Yi Ke Da Xue Xue Bao ; 27(4): 485-7, 2007 Apr.
Artigo em Chinês | MEDLINE | ID: mdl-17545039

RESUMO

OBJECTIVE: To investigate the association between the polymorphism of T(1) locus allele in ADAM33 gene and bronchial asthma in South China Han population. METHODS: Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and DNA sequencing were used to determine the polymorphism of T(1) locus allele in ADAM33 gene in 160 unrelated patients with asthma and 95 unrelated healthy controls from South China Han population. RESULTS: No significant difference was found in T(1) locus allele distribution frequency in populations of UK, US, Germany, Korea, and South China (Chi(2)=9.085, P=0.109). The frequencies of the genotypes (TT, TC, CC) were 80.6% (n=129), 16.9% (n=27) and 2.5% (n=4) in the 160 asthmatic patients and 94.7% (n=90), 3.2% (n=3) and 2.1% (n= 2) in the 95 controls, respectively, showing a significant difference in the distribution of the genotypes (TT, TC, CC ) between asthmatic patients and healthy controls (Chi(2)=10.955, P<0.05). The frequencies of the alleles (T, C) were 0.891 and 0.109 in the asthmatic patients and 0.963 and 0.037 in the controls, respectively, showing also a significant difference in the allele frequency between them (Chi square =8.299, P<0.05). The presence of C allele of ADAM33 gene T1 locus was found to be a greater risk factor in asthmatic patients than in the healthy controls. The odds ratio (OR) of TC and TC+CC were 6.279 (1.849-21.328) and 4.326 (1.620-11.550), respectively, with P value of 0.001 and 0.002, respectively, in comparison with TT genotype. CONCLUSION: The polymorphism of T(1) locus allele in ADAM33 gene is associated with the susceptibility to asthma in South China Han population.


Assuntos
Proteínas ADAM/genética , Asma/genética , Polimorfismo de Fragmento de Restrição , Grupo com Ancestrais do Continente Asiático/genética , China , Frequência do Gene , Predisposição Genética para Doença , Genótipo , Humanos , Análise de Sequência de DNA
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