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Regioselective and enantioselective hydroxylation of propargylic C-H bonds are useful reactions but often lack appropriate catalysts. Here a green and efficient asymmetric hydroxylation of primary and secondary C-H bonds at propargylic positions has been established. A series of optically active propargylic alcohols were prepared with high regio- and enantioselectivity (up to 99% ee) under mild reaction conditions by using P450tol, while the C≡C bonds in the molecule remained unreacted. This protocol provides a green and practical method for constructing enantiomerically chiral propargylic alcohols. In addition, we also demonstrated that the biohydroxylation strategy was able to scaled up to 2.25 mmol scale with the production of chiral propargyl alcohol 2a at a yield of 196 mg with 96% ee, which's an important synthetic intermediate of antifungal drug Ravuconazole.
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OBJECTIVES: To investigate the epidemiological characteristics of respiratory Haemophilus influenzae (HI) infection in children in Suzhou, China and its association with climatic factors and air pollutants. METHODS: The data on air pollutants and climatic factors in Suzhou from January 2016 to December 2019 were collected. Respiratory secretions were collected from 7 940 children with acute respiratory infection who were hospitalized during this period, and bacterial culture results were analyzed for the detection of HI. A stepwise regression analysis was used to investigate the association of HI detection rate with air pollutants (PM2.5, PM10, NO2, SO2, CO, and O3) and climatic factors (monthly mean temperature, monthly mean humidity, monthly total rainfall, monthly total sunshine duration, and monthly mean wind speed). RESULTS: In 2016-2019, the 4-year overall detection rate of HI was 9.26% (735/7 940) among the children in Suzhou. The children aged <1 year and 1-<3 years had a significantly higher HI detection rate than those aged ≥3 years (P<0.01). The detection rate of HI in spring was significantly higher than that in the other three seasons, and the detection rate of HI in autumn was significantly lower than that in the other three seasons (P<0.001). The multiple linear regression analysis showed that PM10 and monthly mean wind speed were independent risk factors for the detection rate of HI: the detection rate of HI was increased by 0.86% for every 10 µg/m3 increase in the concentration of PM10 and was increased by 5.64% for every 1 m/s increase in monthly mean wind speed. Air pollutants and climatic factors had a lag effect on the detection rate of HI. CONCLUSIONS: HI is an important pathogen for acute respiratory infection in children in Suzhou and is prevalent in spring. PM10 and monthly mean wind speed are independent risk factors for the detection rate of HI.
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Poluentes Atmosféricos , Poluição do Ar , Infecções por Haemophilus , Infecções Respiratórias , Criança , Humanos , Poluentes Atmosféricos/efeitos adversos , Poluentes Atmosféricos/análise , Estações do Ano , China/epidemiologia , Infecções por Haemophilus/etiologia , Infecções por Haemophilus/induzido quimicamente , Poluição do Ar/efeitos adversos , Poluição do Ar/análiseAssuntos
Fístula Arteriovenosa , Malformações Vasculares do Sistema Nervoso Central , Embolização Terapêutica , Humanos , Fístula Arteriovenosa/patologia , Malformações Vasculares do Sistema Nervoso Central/complicações , Malformações Vasculares do Sistema Nervoso Central/diagnóstico por imagem , Malformações Vasculares do Sistema Nervoso Central/patologiaRESUMO
Sulfoxaflor is a new systemic insecticide developed as a replacement for older neonicotinoids which are known to be toxic to pollinators. However, its metabolism in nectar and effect on nectar biosynthesis have not been investigated. After soil and foliar application, sulfoxaflor and its main metabolites in soil, leaf and Salvia splendens nectar, were measured by liquid chromatography coupled with triple quadrupole mass spectrometer (LC-MS/MS). The chemical composition between the clean and sulfoxaflor spiked nectar were also compared. The activities of two possible sulfoxaflor detoxifying enzymes in S. splendens nectar, nitrile hydratase and glutathione-s-transferase, were measured by LC-MS and spectrophotometry. S. splendens nectar proteome was investigated by high-resolution orbitrap-based MS/MS to screen for sulfoxaflor detoxifying relevant proteins. S. splendens could absorb sulfoxaflor through root or leaf surface and secrete a proportion of sulfoxaflor along with its metabolites into the nectar. After soil application, sulfoxaflor's low toxic metabolite X11719474 was dominant in the nectar and reached an average concentration of 8905 ppb. However, after foliar application, sulfoxaflor was dominant over its metabolites in the nectar. S. splendens nectar has no nitrile hydratase and glutathione-s-transferase activity and none of the 106 proteins identified in the nectar were predicted to function in detoxifying sulfoxaflor. Soil and foliar sulfoxaflor application can result in different profiles of sulfoxaflor and its metabolites presented in the nectar. However, sulfoxaflor had no effects on S. splendens nectar secretion and chemical composition and cannot be directly detoxified by components in the nectar.
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Inseticidas , Salvia , Cromatografia Líquida , Glutationa , Inseticidas/análise , Inseticidas/toxicidade , Neonicotinoides/análise , Néctar de Plantas/química , Proteoma , Piridinas , Solo/química , Compostos de Enxofre , Espectrometria de Massas em Tandem , TransferasesRESUMO
BACKGROUND AND AIMS: Many angiosperms can secrete both floral (FN) and extrafloral (EFN) nectar. However, much remains unclear about how EFN and FN differ in secretion, composition and ecological function, especially when both FN and EFN are secreted on flowers of the same species. METHODS: Hemerocallis citrina flowers secrete both FN and EFN. The FN and EFN traits including volume, presentation pattern and temporal rhythms of secretion were compared by field observation. Sugar and amino acid contents were analysed using regular biochemical methods, whereas the proteome was investigated by combined gel-based and gel-free approaches. Animal feeders on FN and EFN were investigated by field observation. Hemerocallis citrina plants were exposed by soil drenching to two systemic insecticides, acetamiprid and imidacloprid, and the concentration of these in FN and EFN was measured by ultra-high performance liquid chromatography coupled with mass spectrometry. KEY RESULTS: Hemerocallis citrina FN was concentrated and sucrose dominant, secreted in the mature flower tube and served as a reward for pollinators. Conversely, EFN was hexose rich, more dilute and less rich in sugar and amino acids. EFN was secreted on the outside of developing floral buds, and was likely to attract predatory animals for defence. EFN had fewer phenolics, but more pathogenesis-related components, such as chitinase and glucanase. A significantly different proteomic profile and enzymatic activities between FN and EFN suggest that they had different biosynthesis mechanisms. Both neonicotinoid insecticides examined became present in both nectar types soon after application, but in greater concentration within EFN; EFN also attracted a wider range of insect species than FN. CONCLUSIONS: Hemerocallis citrina FN and EFN differed in production, composition and ecological function. The EFN pathway could be a significant way for neonicotinoids to enter the wild food chain, and must be considered when evaluating the risks to the environment of other systemic insecticides.
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Formigas , Hemerocallis , Inseticidas , Animais , Carboidratos , Flores/metabolismo , Hemerocallis/metabolismo , Neonicotinoides , Néctar de Plantas/metabolismo , Proteômica , AçúcaresRESUMO
Cerebral venous sinus thrombosis is a special cerebrovascular disease affecting young adult and middle-aged people. The clinical manifestations of cerebral venous sinus thrombosis are diverse and nonspecific; thus, imaging plays an important role in early diagnosis. Anticoagulation with heparin is the preferred treatment for cerebral venous sinus thrombosis. Endovascular treatment is also being increasingly used to achieve recanalization of the cerebral venous sinus. We herein describe a woman in her early 50s who was diagnosed with cerebral venous sinus thrombosis for which anticoagulation with heparin was ineffective. To improve her symptoms and prognosis, we selected balloon venoplasty to treat the right sigmoid sinus thrombosis. Her condition subsequently improved, and no recurrence was observed after several follow-ups.
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Trombose dos Seios Intracranianos , Feminino , Heparina/uso terapêutico , Humanos , Pessoa de Meia-Idade , Trombose dos Seios Intracranianos/diagnóstico por imagem , Trombose dos Seios Intracranianos/tratamento farmacológico , Adulto JovemRESUMO
BACKGROUND: Programmed death-ligand 1 (PD-L1) is an important immune checkpoint inhibitor. Recent studies suggest that the PD-L1-mediated pathway may be a promising target in allergic asthma. However, the mechanism by which PD-L1 represses neutrophilic asthma (NA) remains unclear. In this study, we examined correlations between the expression of PD-L1 and the production of T helper cell type 1 (Th1), T helper cell type 2 (Th2), and T helper cell type 17 (Th17) cells in pediatric patients with NA and a mouse model. METHODS: The clinical samples of 26 children with asthma and 15 children with a bronchial foreign body were collected over a period of 12 months by the Children's Hospital of Soochow University. An experimental mouse model of asthma was established to study NA. An enzyme-linked immunoassay (ELISA) was used to assess soluble PD-L1 (sPD-L1) and cytokines [e.g., interleukin (IL)-4, IL-6, interferon gamma (IFN-γ), IL-17 and granulocyte-macrophage colony-stimulating factor (GM-CSF)] in bronchoalveolar lavage fluid (BALF). RESULTS: NA patients had significantly higher levels of sPD-L1, IL-6, IL-17, and GM-CSF in their BALF than non-NA and control patients (P<0.05). In a murine model of asthma, the positive rate and fluorescence intensity of PD-L1 in the NA group and the immunoglobulin G (IgG)-treated NA group were higher than in the PD-L1 antibody (Ab)-treated NA group and the phosphate-buffered saline (PBS) control group (P<0.05). In the plasma and the BALF of the NA group and the IgG-treatment NA group, the levels of IL-17, IL-4, tumor necrosis factor alpha (TNF-α), and granulocyte colony-stimulating were higher than those in the PBS control group (P<0.05). The histopathological examination of lung tissues from all mice groups showed that a large number of inflammatory cells were found around the airway in the NA group and the IgG-treatment group. CONCLUSIONS: PD-L1 may contribute to the Th17/IL-17 immune response, which is associated with neutrophilic inflammation and asthma. A PD-L1 blockade reduces pulmonary neutrophils and mucus production.
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BACKGROUND: This study set out to evaluate the clinical significance and diagnostic effectiveness of serological tests and real-time polymerase chain reactions (RT-PCR) in children of different age groups and disease durations infected with Mycoplasma pneumoniae (MP). METHODS: Pediatric patients with lower respiratory tract infection (LRTI) confirmed by polymerase chain reaction (PCR) were enrolled and subjected to bronchoalveolar lavage fluid PCR (BALF-PCR) for MP infection. The diagnostic values of the serum immunoglobulin M (IgM) test, paired sera immunoglobulin G (IgG) test, RT PCR applied to nasopharyngeal aspirates (NPA-PCR), and combined IgM and NPA-PCR test were evaluated. RESULTS: When BALF PCR was used as the gold standard, the MP positivity rate of combined IgM and NPA PCR was 78.85%in children aged 3-5 years. The positivity rates of IgM, NPA PCR, and combined IgM and NPA PCR in children older than 5 years were 71.21%, 72.72%, and 84.85%, respectively. The detection rate of combined IgM and NPA PCR was consistent with BALF PCR (Kappa =0.727). The MP positivity rates of combined IgM and NPA PCR at 1-2 weeks was as high as 91.11%, and was consistent with the BALF PCR (Kappa =0.756). Moreover, the positivity rates of IgM or NPA PCR at 2-3 weeks were 63.16%, and were consistent with each other (Kappa =0.771). CONCLUSIONS: Combined IgM and NPA PCR is the optimal test to confirm MP infection among children aged 3-5 years in cases with a disease duration of less than2 weeks, and either NPA PCR or IgM is recommended for children older than 5 years with a disease duration of 2-3 weeks. KEYWORDS: Mycoplasma pneumoniae pneumonia (MPP); diagnosis; children; age; disease duration.
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Nasofaringe/patologia , Neurofibromatose 1/cirurgia , Neurofibrossarcoma/cirurgia , Orofaringe/patologia , Pré-Escolar , Dispneia/patologia , Humanos , Masculino , Nasofaringe/cirurgia , Terapia Neoadjuvante , Neurofibromatose 1/patologia , Neurofibrossarcoma/patologia , Orofaringe/cirurgia , Tomografia Computadorizada por Raios XRESUMO
BACKGROUND: Mycoplasma pneumoniae (M. pneumoniae) is one of the most common causes of community-acquired pneumonia in children. Recent studies demonstrated that the incidence of severe or fatal M. pneumoniae was gradually increasing, which may be related to the excessive inflammation. However, the exact pathogenesis of excessive inflammation in Mycoplasma pneumoniae pneumonia(MPP) is still unclear. This study aimed to reveal the role of miR-29c/B7-H3/Th17 axis in children with MPP. METHODS: Children hospitalized in Respiratory Department during Jan. 2014 to Dec. 2015 were enrolled. All children enrolled was confirmed with MP infection using real-time PCR and ELISA. Children were excluded if they were co-infected with other pathogens. A total of 52 children with MPP and 26 controls were enrolled. miR-29c expression in monocytes of children with MPP was determined by real-time PCR and soluble B7-H3 (sB7-H3) and IL-17 were determined by ELISA, and explore their clinical significance. miR-29c overexpression and silencing technology and luciferase reporter assay were performed to confirm whether B7-H3 is the direct target of miR-29c. The levels of transcription factor ROR-γt in CD4+ T cells and cytokine IL-17A in supernatant were detected after stimulated by different concentrations of B7-H3 fusion protein in vitro. RESULTS: Of all 52 children with MPP, the mean age of the children were 77 ± 33 months, and 23 cases were male accounting for 44.2%. Nineteen cases had pleural effusion accounting for 36.5%. Children with MPP had significantly lower level of miR-29c and higher level of sB7-H3 and IL-17 compared to controls (both P < 0.05). The level of miR-29c significantly increased during convalescent phase compared to that of acute phase while sB7-H3 and IL-17 significantly decreased during convalescent phase (both P < 0.05). There was a positive correlation between the level of sB7-H3 and IL-17 in children with MPP during acute-stage (r = 0.361,P = 0.009). Children with MPP combined with pleural effusion had significantly higher level of sB7-H3 compared to those without pleural effusion (9952.3 ± 3065.3 vs. 7449.7 ± 2231.5, pg/ml), and the levels of sB7-H3 was positively correlated with the number of days of fever. The level of miR-29c was negatively correlated with M. pneumoniae specific IgG, IgM level. High concentrations of B7-H3(15µg/ml) could enhance ROR-γt expression and increase IL-17A. Functional studies based on luciferase reporter assay and immunofluorescence staining suggested that B7-H3 is the direct target of miR-29c, and miR-29c silencing or overexpression could up- or down-regulate the expression of B7-H3 in THP-1 cells. CONCLUSIONS: The axis of miR-29c/B7-H3/Th17 plays a vital role in children with MPP through excessive inflammation. miR-29c and B7-H3 may be the new target for the prevention and treatment of MPP, and may be the novel and potential biomarkers for the assessment of prognosis.
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Antígenos B7/metabolismo , Interleucina-17/metabolismo , MicroRNAs/metabolismo , Mycoplasma pneumoniae , Pneumonia por Mycoplasma/diagnóstico , Pneumonia por Mycoplasma/metabolismo , Estudos de Casos e Controles , Pré-Escolar , Infecções Comunitárias Adquiridas , Feminino , Humanos , Lactente , Masculino , Pneumonia por Mycoplasma/etiologiaRESUMO
OBJECTIVE: To explore the symptomatic burden of patients with essential thrombocythemia (ET) and its relation with clinical characteristics including the mutation status, therapeutic protocols and sex. METHODS: Total of 173 Chinese ET patients were selected and grouped on the basis of disease characteristics (mutation status, therapeutic pro to- cols, and sex). RESULTS: All the groups showed low-to-high symptom burden, with the highest in the Hu (hydroxyurea)-group (total symptom score [TSS], 14.7; range, 7.6-14.7). In the JAK2V617F-positive, Hu-treated, and female groups TSS and independent symptom scores were higher than those in the control group. The CALR-positive and IFN-α-treated groups had lower overall and individual scores as compared with groups lacking the corresponding characteristics. As the number of characteristics (JAK2V617F-positive, Hu-treated, and female) increases, the severity of symptoms gradually increased. CONCLUSION: The different characteristics have various effects on symptom burden in ET patients. The accumulation of certain characteristics will lead to more severe symptom burden, thus the patientï¼s symptom burden should be considered comprehensively when making up the treatment schemes and prognosis.
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Trombocitemia Essencial , Povo Asiático , Calreticulina , Feminino , Humanos , Hidroxiureia , Janus Quinase 2 , MutaçãoRESUMO
OBJECTIVE: To study the epidemiological characteristics of respiratory adenovirus (ADV) infections in children from the Suzhou area, China. METHODS: The clinical data of ADV-positive children out of 35 529 children with respiratory tract infections who were hospitalized in the Children's Hospital of Soochow University between January 2006 and December 2015 were retrospectively studied. RESULTS: Of the 35 529 children with respiratory tract infections, 440 (1.24%) were ADV-positive. There was no significant difference in the rate of ADV infections between boys and girls (1.18% vs 1.34%). The ADV infection rates of children at the age of <1 year old, 1-3 years old, 3-7 years old and 7-14 years old were 0.39%â(71/18 002), 1.12%â(103/9 191), 3.14%â(201/6 398), and 3.35%(â65/1 938) respectively and the rate increased with age (P<0.01). The ADV infection rates in spring [1.85%(60/8 658)] and summer [2.20%(189/8 606)] were significantly higher than in autumn [0.30%(27/8 952)] and winter [0.69%(64/9 313)] (P<0.01). CONCLUSIONS: The ADV infection rate is increased with age in the children from the Suzhou area, but it is not associated with gender. ADV infections are more common in spring and summer.
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Infecções por Adenoviridae/epidemiologia , Infecções Respiratórias/epidemiologia , Adolescente , Criança , Pré-Escolar , China/epidemiologia , Feminino , Humanos , Lactente , Masculino , Fatores de TempoRESUMO
OBJECTIVE: To investigate the percentage of T lymphocyte subsets and allergen screening results in infants and young children with Mycoplasma pneumoniae (MP) infection complicated by wheezing. METHODS: Flow cytometry was used to measure the percentage of peripheral blood T cell subsets in 354 infants and young children with MP infection complicated by wheezing (MP wheezing group), 336 infants and young children with MP infection but without wheezing (MP non-wheezing group), and 277 children with recurrent wheezing (recurrent wheezing group). Allergen screening was also performed for these children. RESULTS: Both the MP wheezing group and recurrent wheezing group had significantly lower percentages of CD3+ and CD3+CD8+ lymphocytes than the MP non-wheezing group (p<0.05). The MP groups with or without wheezing had a significantly higher percentage of CD3+CD4+ lymphocytes than the recurrent wheezing group (p<0.05). Both the MP wheezing group and recurrent wheezing group had significantly higher percentages of CD3-CD19+ and CD19+CD23+ lymphocytes than the MP non-wheezing group (p<0.05), and the recurrent wheezing group had the highest percentages (p<0.05). The overall positive rate of food allergens was significantly higher than that of inhaled allergens (30.3% vs 14.7%; p<0.05). The positive rates of food and inhaled allergens in the recurrent wheezing group and MP wheezing group were significantly higher than in the MP non-wheezing group (p<0.05), and the recurrent wheezing group had the highest rates. CONCLUSIONS: Imbalance of T lymphocyte subsets and allergic constitution play important roles in the pathogenesis of MP infection complicated by wheezing in infants and young children.
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Alérgenos/imunologia , Pneumonia por Mycoplasma/complicações , Sons Respiratórios/etiologia , Subpopulações de Linfócitos T/imunologia , Pré-Escolar , Feminino , Humanos , Lactente , Masculino , Pneumonia por Mycoplasma/imunologiaRESUMO
OBJECTIVE: To investigate the bacterial pathogenic characteristics of respiratory tract infection in children. METHODS: The medical data from 14,994 children with respiratory tract infection who were hospitalized in Children's Hospital Affiliated to Soochow University between November 2005 and October 2014 were retrospectively reviewed. RESULTS: Among the 14,994 sputum samples from the children with respiratory tract infection, 3,947 (26.32%) had a positive bacterial culture. The most common bacterial pathogen was Streptococcus pneumonia (12.79%), followed by Haemophilus influenzae (5.02%) and Moraxella catarrhalis (2.91%). The bacterial detection rates differed significantly in different years and seasons and children of different ages (P<0.01). The children who had not taken antibacterial agents before admission had a significantly higher positive bacterial culture rate than those who had taken antibacterial agents (P<0.01). There were significant differences in the bacterial detection rate among the children with different course of disease (<1â month, 1-3â months and >3 months) (P<0.05). The detection rates of Streptococcus pneumonia, Moraxella catarrhalis and Acinetobacter baumannii showed an increased trend with a prolonged disease course (P<0.05). CONCLUSIONS: Streptococcus pneumonia is the most common bacterial pathogen causing respiratory tract infection in children, followed by Haemophilus influenzae and Moraxella catarrhalis. The detection rate of bacterial pathogens varies in different years and seasons and children of different ages. The course of the disease and application of antibacterial agents outside hospital can affect the detection rate of bacterial pathogens in children with respiratory tract infection.
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Bactérias/isolamento & purificação , Infecções Respiratórias/microbiologia , Adolescente , Antibacterianos/uso terapêutico , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Recém-Nascido , Masculino , Estações do Ano , Fatores de TempoRESUMO
OBJECTIVE: To compare the detection rates of Mycoplasma pneumoniae (MP) from nasopharyngeal aspirates (NPA) and bronchoalveolar lavage fluid (BALF) in children with pneumonia. METHODS: A total of 164 hospitalized children with pneumonia were enrolled. NPA and BALF of these children were collected within 24 hours of admission, and MP-DNA was detected by fluorescence quantitative PCR. Venous blood samples of all these children were collected within 24 hours of admission and on days 7-10 of treatment, and serum MP-IgM was detected using ELISA. RESULTS: The positive rate of MP-DNA in NAP of the 164 cases was 51.8% , which was lower than 63.4% as the detection rate of MP-IgM in serum (P=0.044), and the two detection rates were moderately consistent with each other (Kappa=0.618, P<0.01). The positive rate of MP in BALF was 71.3%, which was not significantly different with that of MP-IgM in serum (P>0.05), and the detection rates were well consistent (Kappa=0.793, P<0.01). The detection rate of MP in NPA was lower than that in BALF (P<0.01), with moderate consistency between two of them (Kappa=0.529, P<0.01). The median MP copy number in BALF was significantly higher than that in NPA (P<0.01). The MP detection rates in NPA and BALF were significantly different among different courses of disease (P<0.05). As the course of disease extended, the MP detection rates in both NPA and BALF showed a declining trend; children with MP pneumonia of 1-2 weeks' duration and 2-4 weeks' duration had a higher MP-DNA detection rate in BALF than in NPA (P<0.05). CONCLUSIONS: MP-DNA in BALF has a high sensitivity, with a great significance for early diagnosis of MP pneumonia, while NPA MP-DNA tests may lead to a missed diagnosis.
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Líquido da Lavagem Broncoalveolar/microbiologia , DNA Bacteriano/análise , Nasofaringe/microbiologia , Pneumonia por Mycoplasma/diagnóstico , Adolescente , Criança , Pré-Escolar , Feminino , Humanos , Lactente , MasculinoRESUMO
BACKGROUND: Hepatocellular carcinoma (HCC) is one of the most common malignant tumors. We analyzed the expression of nuclear-transcription factor-kappa B (NF-kappaB) during hepatocarcinogenesis in order to evaluate its dynamic expression and its clinical value in the development and diagnosis of HCC. METHODS: Hepatoma models were induced by oral administration of 2-acetamidoflurene (2-FAA) to male Sprague-Dawley rats. Morphological changes were observed after hematoxylin and eosin staining. The cellular distribution of NF-kappaB expression during different stages of cancer development was investigated by immunohistochemistry, and the level of NF-kappaB expression in liver tissues was quantitatively analyzed by ELISA. The gene fragments of hepatic NF-kappaB were amplified by nested-polymerase chain reaction assay. RESULTS: Hepatocytes showed vacuole-like degeneration during the early stages, then had a hyperplastic nodal appearance during the middle stages, and finally progressed to tubercles of cancerous nests with high differentiation. The NF-kappaB-positive material was buff-colored, fine particles localized in the nucleus, and the incidence of NF-kappaB-positive cells was 81.8% in degeneration, 83.3% in precancerous lesions, and 100% in cancerous tissues. All of these values were higher than those in controls (P<0.01). Hepatic NF-kappaB expression and hepatic NF-kappaB-mRNA were also higher during the course of HCC development (P<0.01). CONCLUSION: The NF-kappaB signal transduction pathway is activated during the early stages of HCC development, and its abnormal expression may be associated with the occurrence of HCC.
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Biomarcadores Tumorais/metabolismo , Carcinoma Hepatocelular/metabolismo , Progressão da Doença , Neoplasias Hepáticas/metabolismo , NF-kappa B/metabolismo , 2-Acetilaminofluoreno/efeitos adversos , Animais , Carcinoma Hepatocelular/induzido quimicamente , Carcinoma Hepatocelular/patologia , Transformação Celular Neoplásica/metabolismo , Transformação Celular Neoplásica/patologia , Modelos Animais de Doenças , Hepatócitos/metabolismo , Hepatócitos/patologia , Fígado/metabolismo , Fígado/patologia , Neoplasias Hepáticas/induzido quimicamente , Neoplasias Hepáticas/patologia , Masculino , Ratos , Ratos Sprague-Dawley , Transdução de SinaisRESUMO
BACKGROUND: Hepatic hypoxia-inducible factor-1 (HIF-1) is activated in the progression of hepatocellular carcinoma (HCC). This study aimed to investigate the dynamic alterations of HIF-1alpha and its gene expression so as to explore the relationship between HIF-1alpha expression and hepatocarcinogenesis at the early stage of HCC. METHODS: A hepatoma model was made with 2-fluorenyl-acetamide (2-FAA) in male Sprague-Dawley rats. Morphological changes of rat hepatocytes were assessed pathologically (HE staining). The dynamic expression of hepatic and circulating HIF-1alpha was quantitatively analyzed by ELISA. The gene fragments of hepatic HIF-1alpha mRNA were amplified by RT-PCR and confirmed by sequencing. The cellular distribution of hepatic HIF-1alpha expression was confirmed by immunohistochemistry. RESULTS: Histological examination confirmed granule-like degeneration to atypical hyperplasia and HCC development in rat hepatocytes and progressive increases in the levels of hepatic and circulating HIF-1alpha and its gene expression during the course. The levels of HIF-1alpha expression in the liver and blood of rats with hepatoma were significantly higher than those in normal rats and those with degeneration. Immunohistochemical analysis confirmed the positive expression and hepatocyte distribution of HIF-1alpha in the development of rat hepatoma. A positive relationship was found between HIF-1alpha expression in the liver and blood (P<0.01). CONCLUSIONS: The above observations support the hypothesis that the overexpression of HIF-1alpha and its gene are closely associated with the malignant transformation of hepatocytes and play an important role at the stage of hepatocarcinogenesis.
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Carcinoma Hepatocelular/metabolismo , Transformação Celular Neoplásica/metabolismo , Regulação Neoplásica da Expressão Gênica , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Neoplasias Hepáticas/metabolismo , Lesões Pré-Cancerosas/metabolismo , 2-Acetilaminofluoreno , Animais , Sequência de Bases , Carcinoma Hepatocelular/induzido quimicamente , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/patologia , Transformação Celular Neoplásica/induzido quimicamente , Transformação Celular Neoplásica/genética , Transformação Celular Neoplásica/patologia , Modelos Animais de Doenças , Ensaio de Imunoadsorção Enzimática , Subunidade alfa do Fator 1 Induzível por Hipóxia/sangue , Subunidade alfa do Fator 1 Induzível por Hipóxia/genética , Imuno-Histoquímica , Fígado/metabolismo , Fígado/patologia , Neoplasias Hepáticas/induzido quimicamente , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/patologia , Masculino , Dados de Sequência Molecular , Lesões Pré-Cancerosas/induzido quimicamente , Lesões Pré-Cancerosas/genética , Lesões Pré-Cancerosas/patologia , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Regulação para CimaAssuntos
Inibidores da Angiogênese/uso terapêutico , Angiopoietina-2 , Carcinoma Hepatocelular/patologia , Neoplasias Hepáticas/patologia , Neovascularização Patológica/prevenção & controle , Angiopoietina-2/antagonistas & inibidores , Angiopoietina-2/genética , Angiopoietina-2/metabolismo , Carcinoma Hepatocelular/irrigação sanguínea , Carcinoma Hepatocelular/metabolismo , Células Endoteliais/efeitos dos fármacos , Células Endoteliais/metabolismo , Regulação Neoplásica da Expressão Gênica , Humanos , Neoplasias Hepáticas/irrigação sanguínea , Neoplasias Hepáticas/metabolismo , Estrutura Molecular , Interferência de RNA , Transdução de Sinais/efeitos dos fármacos , Fator A de Crescimento do Endotélio Vascular/antagonistas & inibidores , Fator A de Crescimento do Endotélio Vascular/metabolismoRESUMO
OBJECTIVE: To investigate the kinetic expression and alteration of nuclear transcription factor-kappa B (NF-kappaB) and its gene in hepatocellular carcinoma (HCC) development. METHODS: A hepatoma model was established with N-(2-fluorenyl) acetamide (2-FAA) using male SD rats. Morphological changes and dynamic alterations of NF-kappaB and NF-kappaB mRNA of the rat livers at different stages of HCC development were observed by pathological examinations. The liver specimens from HCC patients were collected by self-control method. The expression of NF-kappaB was quantitatively analyzed by ELISA. RESULTS: Hepatocytes showed vacuole-like denaturation, atypical hyperplasia, and transformation into highly differentiated cancerous hepatocytes with increasing tendencies of liver NF-kappaB and NF-kappaB mRNA expressions. The NF-kappaB positive material was granule-like and stained brown, with dot-nest-like staining localized in the nuclei and cytoplasm of HCC cells, but only in the cytoplasm of the cells of park cancer tissues. Its expression in HCC cells was stronger than that in their surrounding tissues (chi2 = 13.1, P less than 0.01). No positive relationship was found between NF-kappaB expression and histological grades, the number of tumors, or size of the tumors. CONCLUSION: The expression of NF-kappaB and its gene are associated with the development of HCC. To inhibit the expression may be useful to HCC therapy.