RESUMO
Legionellosis caused by Legionella longbeachae is diagnosed mainly by PCR. We report a case of L. longbeachae infection in mainland China, which was diagnosed by metagenomic next-generation sequencing, in a man who developed an epileptic seizure after using moxifloxacin. Metagenomic next-generation sequencing may be a useful tool to detect Legionella spp.
Assuntos
Epilepsia/induzido quimicamente , Legionelose , Moxifloxacina/efeitos adversos , Pneumonia Bacteriana/tratamento farmacológico , Convulsões/induzido quimicamente , China , Humanos , Legionella longbeachae , Legionelose/tratamento farmacológico , Masculino , Pessoa de Meia-Idade , Moxifloxacina/uso terapêuticoRESUMO
Long non-coding RNAs (lncRNAs) are emerging as important regulators involved in the pathogenesis of many diseases. However, it is still unknown if they contribute to the occurrence of acute pancreatitis (AP). Here, we identified a lncRNA CASC2 (Cancer Susceptibility Candidate 2) was significantly upregulated in the pancreatic tissues from AP patients. Knockdown or overexpression of CASC2 in vitro could specifically repress or induce the expression of two proinflammatory cytokines including IL6 (Interleukin 6) and IL17, respectively. Changing the expression levels of several transcription factors that were predicted to bind to the promoter of CASC2, we found c-MYC could specifically regulate the expression of CASC2. Using immunoprecipitation, mass spectrometry, and co-immunoprecipitation assays, we proved that c-MYC assembled a transcriptional complex with PCAF (p300/CBP-associated Factor) and CtBP1/2 (C-terminal Binding Protein 1 and 2), terming as the CtBP-PCAF-c-MYC (CPM) complex. Further investigation revealed that CtBPs were amplified in the pancreatic tissues from AP patients and they functioned as coactivators to induce the expression of CASC2 and thus led to the upregulation of IL6 and IL17. Moreover, we identified that decreased DNA methylation levels in the promoters of CtBPs and inflammatory stimuli coactivated the expression of CtBPs. Collectively, we identified a new signaling pathway in which DNA methylation and inflammatory stimuli coregulate the CPM complex to activate CASC2 expression, whose induction further activates the expression of IL6 and IL17, eventually aggravating inflammation response and causing the pathology of AP.
Assuntos
Oxirredutases do Álcool/metabolismo , Metilação de DNA , Proteínas de Ligação a DNA/metabolismo , Pancreatite/metabolismo , Proteínas Proto-Oncogênicas c-myc/metabolismo , Proteínas Supressoras de Tumor/metabolismo , Fatores de Transcrição de p300-CBP/metabolismo , Doença Aguda , Oxirredutases do Álcool/genética , Citocinas/genética , Citocinas/metabolismo , Proteínas de Ligação a DNA/genética , Regulação da Expressão Gênica , Humanos , Neoplasias Pancreáticas/metabolismo , Pancreatite/sangue , Pancreatite/genética , Proteínas Proto-Oncogênicas c-myc/genética , Proteínas Supressoras de Tumor/genética , Fatores de Transcrição de p300-CBP/genéticaRESUMO
Nasopharyngeal carcinoma (NPC) is highly prevalent in Southeast Asia, and an unfavorable outcome is usually attributed to advanced stage NPC. Current methods for the early diagnosis of NPC have limitations in clinical practice. The aim of this study was to investigate the diagnostic ability of Septin 9 methylation for NPC. A quantitative methylation-sensitive PCR (qMS-PCR) assay was developed to measure the methylation status and levels of Septin 9 in nasopharyngeal tissues and paired swabs from patients with NPC, chronic nasopharyngitis, and healthy donors. Methylated Septin 9 was detected in 92% (23/25) of NPC tissues and 25% (4/16) of nasopharyngitis controls (p < 0.05). High-frequency hypermethylation with decreased mRNA expression of Septin 9 in NPC was also identified. Further, Septin 9 methylation was identified in 90.5% (19/21) of NPC biopsies and 71.4% (15/21) of paired swabs, indicating a good concordance between the two sample types. In addition, methylated Septin 9 was found in 16 (72.7%) nasal swabs from 22 NPC patients, 2 of 19 (10.5%) nasopharyngitis, but not in any of the healthy controls (p < 0.01). The methylation score in nasal swabs of the NPC group was also significantly higher than that of non-NPC controls (p < 0.001). Moreover, receiver operating characteristic (ROC) curve analysis showed an area under the curve (AUC) of 0.882 of Septin 9 methylation tests to discriminate NPC from non-NPC subjects. Our study demonstrated that frequent methylation of Septin 9 was present in NPC. Its detection in nasopharyngeal swabs may provide a minimally invasive and informative method for identifying early NPC cases.
Assuntos
Epigênese Genética , Carcinoma Nasofaríngeo/diagnóstico , Neoplasias Nasofaríngeas/diagnóstico , Nasofaringite/diagnóstico , RNA Mensageiro/genética , Septinas/genética , Área Sob a Curva , Estudos de Casos e Controles , Metilação de DNA , Diagnóstico Diferencial , Detecção Precoce de Câncer/métodos , Humanos , Carcinoma Nasofaríngeo/genética , Carcinoma Nasofaríngeo/metabolismo , Carcinoma Nasofaríngeo/patologia , Neoplasias Nasofaríngeas/genética , Neoplasias Nasofaríngeas/metabolismo , Neoplasias Nasofaríngeas/patologia , Nasofaringite/genética , Nasofaringite/metabolismo , Nasofaringite/patologia , Nasofaringe/metabolismo , Nasofaringe/patologia , Estadiamento de Neoplasias , Reação em Cadeia da Polimerase/métodos , RNA Mensageiro/metabolismo , Curva ROC , Septinas/metabolismoRESUMO
BACKGROUND: It is challenging to distinguish intestinal tuberculosis from Crohn's disease due to dynamic changes in epidemiology and similar clinical characteristics. Recent studies have shown that polymorphisms in genes involved in the interleukin (IL)-23/IL-17 axis may affect intestinal mucosal immunity by affecting the differentiation of Th17 cells. AIM: To investigate the specific single-nucleotide polymorphisms (SNPs) in genes involved in the IL-23/IL-17 axis and possible pathways that affect susceptibility to intestinal tuberculosis and Crohn's disease. METHODS: We analysed 133 patients with intestinal tuberculosis, 128 with Crohn's disease, and 500 normal controls. DNA was extracted from paraffin-embedded specimens or whole blood. Four SNPs in the IL23/Th17 axis (IL22 rs2227473, IL1ß rs1143627, TGFß rs4803455, and IL17 rs8193036) were genotyped with TaqMan assays. The transcriptional activity levels of different genotypes of rs2227473 were detected by dual luciferase reporter gene assay. The expression of IL-22R1 in different intestinal diseases was detected by immunohistochemistry. RESULTS: The A allele frequency of rs2227473 (P = 0.030, odds ratio = 0.60, 95% confidence interval: 0.37-0.95) showed an abnormal distribution between intestinal tuberculosis and healthy controls. The presence of the A allele was associated with a higher IL-22 transcriptional activity (P < 0.05). In addition, IL-22R1 was expressed in intestinal lymphoid tissues, especially under conditions of intestinal tuberculosis, and highly expressed in macrophage-derived Langhans giant cells. The results of immunohistochemistry showed that the expression of IL-22R1 in patients with Crohn's disease and intestinal tuberculosis was significantly higher than that in patients with intestinal polyps and colon cancer (P < 0.01). CONCLUSION: High IL-22 expression seems to be a protective factor for intestinal tuberculosis. IL-22R1 is expressed in Langhans giant cells, suggesting that the IL-22/IL-22R1 system links adaptive and innate immunity.
Assuntos
Doença de Crohn/diagnóstico , Células Gigantes de Langhans/patologia , Interleucinas/genética , Receptores de Interleucina/metabolismo , Tuberculose Gastrointestinal/diagnóstico , Adulto , Biópsia , Estudos de Casos e Controles , Doença de Crohn/genética , Doença de Crohn/imunologia , Diagnóstico Diferencial , Feminino , Predisposição Genética para Doença , Células Gigantes de Langhans/imunologia , Humanos , Interleucinas/imunologia , Mucosa Intestinal/citologia , Mucosa Intestinal/imunologia , Mucosa Intestinal/patologia , Masculino , Pessoa de Meia-Idade , Polimorfismo de Nucleotídeo Único , Regiões Promotoras Genéticas/genética , Receptores de Interleucina/imunologia , Fatores de Risco , Tuberculose Gastrointestinal/genética , Tuberculose Gastrointestinal/imunologia , Adulto Jovem , Interleucina 22RESUMO
BACKGROUND: Because of the similarity of intestinal tuberculosis and Crohn's disease in disease phenotype, differential diagnosis has always been a clinical problem. Arachidonic acid metabolites play an important role in the inflammatory response of intestinal tuberculosis and Crohn's disease. Recent studies have shown that the polymorphism locus in the promoter region of LTA4H gene affects LTB4 expression level and the susceptibility to extrapulmonary tuberculosis. Thus, we identified a total of 148 patients with intestinal tuberculosis, 145 with Crohn's disease, and 700 normal controls in this study. METHODS: All the study participants were local Han people from Jiangxi Province in the past eleven years. DNA was extracted from the paraffin-embedded specimens or the whole blood. The LTA4H promoter SNP (rs17525495) was genotyped with TaqMan assay. RESULTS: The T-alleles frequency was not significantly increased in patients with intestinal tuberculosis compared with healthy control group (p=0.630; OR=1.07; 95%CI=0.81-1.41), while patients with Crohn's disease have significantly increased T allele frequency compared with healthy population (p=0.032; OR=1.34; 95%CI=1.03-1.75). During treatment, the presence of the T allele significantly increased the proportion of Crohn's patients requiring glucocorticoids (p<0.05). CONCLUSIONS: The T allele of LTA4H gene SNP (rs17525495) is a risk factor for Crohn's disease instead of intestinal tuberculosis. More importantly, there may be a potential association of the different genotypes of rs17525495 with the treatment efficacy of 5-ASA and glucocorticoids in patients with Crohn's disease. The association between LTA4H polymorphism and drugs therapeutic effects might contribute to the practice of precision medicine and the prediction of clinical outcomes.
Assuntos
Povo Asiático/genética , Doença de Crohn/genética , Epóxido Hidrolases/genética , Etnicidade/genética , Estudos de Associação Genética , Predisposição Genética para Doença , Polimorfismo de Nucleotídeo Único/genética , Tuberculose Gastrointestinal/genética , Adulto , Doença de Crohn/enzimologia , Feminino , Frequência do Gene/genética , Humanos , Masculino , Modelos Genéticos , Tuberculose Gastrointestinal/enzimologiaRESUMO
Six new steroidal saponins, namely glauco-chinaosides A-F, and one known compound were isolated from the tubers of Smilax glauco-china. Their structures were elucidated by a combination of spectroscopic analysis and hydrolysis followed by spectral and chromatographic analysis. Compounds 1-7 were tested in vitro for their cytotoxic activities against four human tumor cell lines (SH-SY5Y, SGC-7901, HCT-116, and Lovo). Compounds 1, 2, and 5 exhibited cytotoxic activity against SGC-7901, with IC50 values of 2.7, 11.5, and 6.8 µM, respectively.
Assuntos
Antineoplásicos Fitogênicos/isolamento & purificação , Antineoplásicos Fitogênicos/farmacologia , Medicamentos de Ervas Chinesas/isolamento & purificação , Medicamentos de Ervas Chinesas/farmacologia , Glicosídeos/isolamento & purificação , Glicosídeos/farmacologia , Saponinas/isolamento & purificação , Saponinas/farmacologia , Smilax/química , Esteróis/isolamento & purificação , Esteróis/farmacologia , Antineoplásicos Fitogênicos/química , Ensaios de Seleção de Medicamentos Antitumorais , Medicamentos de Ervas Chinesas/química , Glicosídeos/química , Células HCT116 , Humanos , Concentração Inibidora 50 , Estrutura Molecular , Ressonância Magnética Nuclear Biomolecular , Rizoma/química , Saponinas/química , Esteróis/químicaRESUMO
Spontaneous neuronal activity in dorsal cochlear nucleus (DCN) may be involved in the physiological processes underlying salicylate-induced tinnitus. As a neuronal activity marker, immediate-early gene (IEG) expression, especially activity-dependent cytoskeletal protein (Arc/Arg3.1) and the early growth response gene-1 (Egr-1), appears to be highly correlated with sensory-evoked neuronal activity. However, their relationships with tinnitus induced by salicylate have rarely been reported in the DCN. In this study, we assessed the effect of acute and chronic salicylate treatment on the expression of N-methyl D-aspartate receptor subunit 2B (NR2B), Arg3.1, and Egr-1. We also observed ultrastructural alterations in the DCN synapses in an animal model of tinnitus. Levels of mRNA and protein expression of NR2B and Arg3.1 were increased in rats that were chronically administered salicylate (200 mg/kg, twice daily for 3, 7, or 14 days). These levels returned to baseline 14 days after cessation of treatment. However, no significant changes were observed in Egr-1 gene expression in any groups. Furthermore, rats subjected to long-term salicylate administration showed more presynaptic vesicles, thicker and longer postsynaptic densities, and increased synaptic interface curvature. Alterations of Arg3.1 and NR2B may be responsible for the changes in the synaptic ultrastructure. These changes confirm that salicylate can cause neural plasticity changes at the DCN level.
Assuntos
Núcleo Coclear/metabolismo , Regulação da Expressão Gênica , Genes Precoces/genética , RNA Mensageiro/genética , Zumbido/genética , Animais , Núcleo Coclear/ultraestrutura , Modelos Animais de Doenças , Imuno-Histoquímica , Masculino , Microscopia Eletrônica de Transmissão , Ratos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase em Tempo Real , Salicilato de Sódio/toxicidade , Sinapses/genética , Sinapses/metabolismo , Zumbido/induzido quimicamente , Zumbido/metabolismoRESUMO
AIM: To investigate pim-3 expression in hepatic stellate cells (HSCs) stimulated by lipopolysaccharide (LPS), and its protective effect on HSCs. METHODS: Rat HSC-T6 cells were stimulated by LPS. The effect of LPS on proliferation and apoptosis of HSC-T6 cells was investigated by methyl thiazoyltetrazolium (MTT) assay and flow cytometry after annexin V-fluorescein isothiocyanate/propidium iodide double staining. pim-3 mRNA and protein were detected by reverse transcriptase polymerase chain reaction and Western blotting at 48 h when HSC-T6 cells were stimulated with 1 µg/mL LPS for 0, 3, 6, 12, 24 and 48 h. The cells without stimulation served as controls. To study the effect of pim-3 kinase on HSC-T6 cells, si-pim3 (siRNA against pim-3) was transfected into HSC-T6 cells. HSC-T6 cells were subjected to different treatments, including LPS, si-pim3, or si-pim3 plus LPS, and control cells were untreated. Protein expression of pim-3 was detected at 48 h after treatment, and cell proliferation at 24 and 48 h by MTT assay. Apoptosis was detected by flow cytometry, and confirmed with caspase-3 activity assay. RESULTS: LPS promoted HSC-T6 cell proliferation and protected against apoptosis. Significantly delayed upregulation of pim-3 expression induced by LPS occurred at 24 and 48 h for mRNA expression (pim-3/ß-actin RNA, 24 or 48 h vs 0 h, 0.81 ± 0.20 or 0.78 ± 0.21 vs 0.42 ± 0.13, P < 0.05), and occurred at 12 h and peaked at 24 and 48 h for protein expression (pim-3/GAPDH protein, 12, or 24 or 48 h vs 0 h, 0.68 ± 0.12, 1.47 ± 0.25 or 1.51 ± 0.23 vs 0.34 ± 0.04, P < 0.01). pim-3 protein was ablated by si-pim3 and upregulated by LPS in HSC-T6 cells at 48 h after treatment (pim-3/GAPDH: si-pim3, si-pim3 plus LPS or LPS vs control, 0.11 ± 0.05, 0.12 ± 0.05 or 1.08 ± 0.02 vs 0.39 ± 0.03, P < 0.01). Ablation of pim-3 by si-pim3 in HSC-T6 cells partly abolished proliferation (OD at 24 h, si-pim3 group or si-pim3 plus LPS vs control, 0.2987 ± 0.050 or 0.4063 ± 0.051 vs 0.5267 ± 0.030, P < 0.01; at 48 h 0.4634 ± 0.056 or 0.5433 ± 0.031 vs 0.8435 ± 0.028, P < 0.01; si-pim3 group vs si-pim3 plus LPS, P < 0.01 at 24 h and P < 0.05 at 48 h), and overexpression of pim-3 in the LPS group increased cell proliferation (OD: LPS vs control, at 24 h, 0.7435 ± 0.028 vs 0.5267 ± 0.030, P < 0.01; at 48 h, 1.2136 ± 0.048 vs 0.8435 ± 0.028, P < 0.01). Ablation of pim3 with si-pim3 in HSC-T6 cells aggravated apoptosis (si-pim3 or si-pim3 plus LPS vs control, 42.3% ± 1.1% or 40.6% ± 1.3% vs 16.8% ± 3.3%, P < 0.01; si-pim3 vs si-pim3 plus LPS, P > 0.05), and overexpression of pim-3 in the LPS group attenuated apoptosis (LPS vs control, 7.32% ± 2.1% vs 16.8% ± 3.3%, P < 0.05). These results were confirmed by caspase-3 activity assay. CONCLUSION: Overexpression of pim-3 plays a protective role in LPS-stimulated HSC-T6 cells.
Assuntos
Células Estreladas do Fígado/efeitos dos fármacos , Lipopolissacarídeos/farmacologia , Animais , Apoptose/efeitos dos fármacos , Caspase 3/metabolismo , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Regulação Enzimológica da Expressão Gênica , Células Estreladas do Fígado/enzimologia , Proteínas Serina-Treonina Quinases/genética , Proteínas Serina-Treonina Quinases/metabolismo , Interferência de RNA , RNA Mensageiro/metabolismo , Ratos , Fatores de Tempo , Transfecção , Regulação para CimaRESUMO
This study investigates the effect of tone inventories on brain activities underlying pitch without focal attention. We find that the electrophysiological responses to across-category stimuli are larger than those to within-category stimuli when the pitch contours are superimposed on nonspeech stimuli; however, there is no electrophysiological response difference associated with category status in speech stimuli. Moreover, this category effect in nonspeech stimuli is stronger for Cantonese speakers. Results of previous and present studies lead us to conclude that brain activities to the same native lexical tone contrasts are modulated by speakers' language experiences not only in active phonological processing but also in automatic feature detection without focal attention. In contrast to the condition with focal attention, where phonological processing is stronger for speech stimuli, the feature detection (pitch contours in this study) without focal attention as shaped by language background is superior in relatively regular stimuli, that is, the nonspeech stimuli. The results suggest that Cantonese listeners outperform Mandarin listeners in automatic detection of pitch features because of the denser Cantonese tone system.
Assuntos
Potenciais Evocados , Idioma , Percepção da Altura Sonora , Percepção da Fala/fisiologia , Estimulação Acústica/métodos , Adulto , Atenção , Comportamento , China , Processamento Eletrônico de Dados , Eletrofisiologia , Feminino , Humanos , Masculino , Reprodutibilidade dos Testes , Processamento de Sinais Assistido por Computador , Fala , Acústica da Fala , Adulto JovemRESUMO
Aspirin (salicylate), as a common drug that is frequently used for long-term treatment in a clinical setting, has the potential to cause reversible tinnitus. However, few reports have examined the inflammatory cytokines expression and alteration of synaptic ultrastructure in the cochlear nucleus (CN) in a rat model of tinnitus. The tinnitus-like behavior of rats were detected by the gap prepulse inhibition of acoustic startle (GPIAS) paradigm. We investigated the expression levels of the tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), N-methyl D-aspartate receptor subunit 2A (NR2A) mRNA and protein in the CN and compared synapses ultrastructure in the CN of tinnitus rats with normal ones. GPIAS showed that rats with long-term administration of salicylate were experiencing tinnitus, and the mRNA and protein expression levels of TNF-α and NR2A were up-regulated in chronic treatment groups, and they returned to baseline 14 days after cessation of treatment. Furthermore, compared to normal rats, repetitive salicylate-treated rats showed a greater number of presynaptic vesicles, thicker and longer postsynaptic densities, increased synaptic interface curvature. These data revealed that chronic salicylate administration markedly, but reversibly, induces tinnitus possibly via augmentation of the expression of TNF-α and NR2A and cause changes in synaptic ultrastructure in the CN. Long-term administration of salicylate causes neural plasticity changes at the CN level.
Assuntos
Núcleo Coclear/metabolismo , Mediadores da Inflamação/metabolismo , Ácido Salicílico , Sinapses/metabolismo , Zumbido/induzido quimicamente , Animais , Comportamento Animal , Núcleo Coclear/imunologia , Núcleo Coclear/ultraestrutura , Modelos Animais de Doenças , Regulação da Expressão Gênica , Interleucina-6/genética , Interleucina-6/metabolismo , Masculino , Plasticidade Neuronal , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Receptores de N-Metil-D-Aspartato/genética , Receptores de N-Metil-D-Aspartato/metabolismo , Reflexo de Sobressalto , Sinapses/imunologia , Sinapses/ultraestrutura , Fatores de Tempo , Zumbido/genética , Zumbido/imunologia , Zumbido/patologia , Zumbido/psicologia , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/metabolismoAssuntos
Imunoglobulina G/metabolismo , Doenças Linfáticas/imunologia , Doenças Linfáticas/patologia , Idoso , Hiperplasia do Linfonodo Gigante/imunologia , Hiperplasia do Linfonodo Gigante/patologia , Diagnóstico Diferencial , Humanos , Doenças Linfáticas/cirurgia , Linfoma/patologia , Masculino , Plasmócitos/imunologia , Pseudolinfoma/imunologia , Pseudolinfoma/patologiaRESUMO
OBJECTIVE: To investigate the value of incidental focal (18)F-FDG uptake in the colon and rectum and characteristics of functional anatomic form for differential diagnosis of colorectal benign or malignant diseases. METHODS: Clinical data and images of incidental focal hypermetabolism focus in colon and rectum of 37 individuals undergoing (18)F-FDG PET-CT were analyzed retrospectively. According to the eventual outcomes of pathological examination and clinical follow-up, these cases were divided into four subgroups: malignant disease, benign tumor (including precancerous change), inflammation and physiological uptake. Radioactive uptake level (SUVmax) and change of delayed imaging (RI) of focal hypermetabolism focus were compared between groups. The data analysis was performed using variance analysis. RESULTS: The average SUVmax was 6.3±3.7, 8.8±6.5, 5.2±1.4, and 3.8±0.9 in malignant disease (n=11), benign (precancerous) tumor (n=9), inflammation (n=9) and physiological uptaking (n=8) respectively. The average SUVmax was 7.6±5.6 in benign and malignant tumor, and 4.7±1.5 in inflammation and physiological uptake. The distinction of average SUVmax was not statistically significant between benign and malignant tumor or inflammation and physiological uptake. But it was higher in tumors as compared to inflammation or physiological uptake with a statistically difference (P<0.05). The RI was 0.3±0.2, 0.4±0.1, 0.3±0.2, 0.4±0.2 in above 4 groups respectively, and the differences were not statistically significant. CONCLUSIONS: The incidental focal hypermetabolism focus in the colon the rectum during (18)F-FDG PET-CT may indicate potential colorectal malignant diseases and precancerous lesions. SUVmax value in focal hypermetabolism focus in the colon and rectum can help to distinguish tumor from inflammation or physiological uptake. But there is no diagnostic value for distinguishing malignant disease from benign tumor.
Assuntos
Doenças do Colo/diagnóstico por imagem , Tomografia por Emissão de Pósitrons/métodos , Doenças Retais/diagnóstico por imagem , Adulto , Idoso , Idoso de 80 Anos ou mais , Diagnóstico Diferencial , Feminino , Fluordesoxiglucose F18 , Humanos , Masculino , Pessoa de Meia-Idade , Estudos RetrospectivosRESUMO
OBJECTIVE: To evaluate the relationship between the Mandarin acceptable noise level (ANL) and the personality trait for normal-hearing adults. METHODS: Eighty-five Mandarin speakers, aged from 21 to 27, participated in this study. ANL materials and the Eysenck Personality Questionnaire (EPQ) questionnaire were used to test the acceptable noise level and the personality trait for normal-hearing subjects. SPSS 17.0 was used to analyze the results. RESULTS: ANL were (7.8 ± 2.9) dB in normal hearing participants. The P and N scores in EPQ were significantly correlated with ANL (r = 0.284 and 0.318, P < 0.01). No significant correlations were found between ANL and E and L scores (r = -0.036 and -.167, P > 0.05). CONCLUSION: Listeners with higher ANL were more likely to be eccentric, hostile, aggressive, and instabe, no ANL differences were found in listeners who were different in introvert-extravert or lying.
Assuntos
Limiar Auditivo , Ruído , Personalidade , Adulto , Audiometria da Fala , Feminino , Humanos , Masculino , Adulto JovemRESUMO
OBJECTIVE: To study the expressions of gastric mucosal proteins in chronic gastritis (CG) rats of Pi-Wei damp-heat syndrome (PWDHS), to investigate the pathogenesis correlated to CG rats of PWDHS, to observe the differential expressions of gastric mucosal proteins in CG rats of PWDHS, and to investigate the mechanisms of Sanren Decoction (SD) for treating CG rats of PWDHS. METHODS: Totally 36 male SD rats were adaptable fed for 3 days and randomly divided into 3 groups, i.e., the normal control group, the CG of PWDHS rat model group (abbreviated as the model group), and the SD treatment group, 12 in each group. The CG of PWDHS rat model was prepared by composite factors. The gastric mucosal protein was separated using two-dimensional gel electrophoresis technique, and stained by Coomassie brilliant blue. The protein spots expressed differently were analyzed by PDquest 8.0 software. The protein spots expressed differently was identified by MALDI-TOF/TOF-MS. RESULTS: The protein spots were 1 025 +/- 3 9, 994 +/- 51, 1 087 +/- 33 in the normal control group, the model group, and the SD treatment group respectively detected from two-dimensional gel electrophoresis profiles. Compared with the normal control group, there were 74 protein spots differentially expressed in the model group, 30 spots up-regulated and 44 spots down-regulated. Compared with the model group, there were 75 protein spots differentially expressed in the SD treatment group, 49 spots up-regulated and 26 spots down-regulated. Five protein spots differentially expressed were successfully identified, i.e., heat shock protein 72 (HSP72), heat shock protein 60 (HSP60), protein disulfide-isomerase (PDI), malate dehydrogenase (MDH), and unnamed protein. CONCLUSIONS: The pathogenesis of CG of PWDHS may be correlated to energy metabolism disturbance and stress. The mechanisms of SD for treating it may possibly adjust differential expressions of gastric mucosal proteins.
Assuntos
Medicamentos de Ervas Chinesas/uso terapêutico , Gastrite/tratamento farmacológico , Gastrite/metabolismo , Fitoterapia , Proteoma/metabolismo , Animais , Mucosa Gástrica/metabolismo , Gastrite/diagnóstico , Masculino , Medicina Tradicional Chinesa , Ratos , Ratos Sprague-DawleyRESUMO
OBJECTIVE: To explore the significance of colonic epithelial cell apoptosis and tumor necrosis factor α (TNF-α) changing in pathogenesis of melanosis coli (MC) in guinea pig and the molecular mechanism of rhubarb (Rhu) in inducing the disease, by means of using different dosages of Rhu to induce the disease. METHODS: One hundred and forty-four male guinea pigs, clean grade, were randomized according to their body weight into 5 groups, the untreated normal group and the 4 Rhu groups treated, respectively, with different doses of Rhu, 3 g/kg·d for low dose (Rhu-l) group, 6 g/kg·d for moderate dose (Rhu-m) group, 12 g/kg·d for high dose (Rhu-h) group and 24 g/kg·d for super-high dose (Rhu-s) group via gastric infusion. All animals were sacrificed 60 days later, their viscera were taken for observing the pathologic and morphologic changes with HE, melanin and melatonin staining, and the apoptosis of colonic epithelial cells was detected with TUNEL stain and transmission electric microscopy. In addition, the levels of TNF-α in serum and colonic tissue were measured using ELISA and RT-PCR. RESULTS: The pathological changes of MC could be found by naked eye in all Rhu groups, especially apparent at caecum and proximal end of colon, but did not found in gallbladder, jejunum and ileum. In normal guinea pigs, the colonic membrane was pink in color with no apparent pigment deposition. Membranous color deepened in the Rhu groups depending on the dosage of Rhu used. MC scoring showed the highest scores revealed in the Rhu-s group (6.00±0.00), which was significantly different to those in the Rhu-l (3.86±0.69), Rhu-m (4.43±0.79) and Rhu-h groups (4.88±0.35, all P<0.05). Levels of cell apoptosis in colon and TNF-α in serum in all Rhu groups were higher than those in the normal group (P<0.01), but showed no significant difference among the Rhu groups (P>0.05). Moreover, a positive correlation was found in the degree of induced MC with apoptosis rate and TNF-α level. CONCLUSIONS: Rhu (anthraquinone purgatives) had apparent effect on inducing MC; its molecular mechanism is maybe to destroy intestinal mucosal barrier and advance proinflammatory factor TNF-α releasing, which leads to colonic epithelial cells apoptosis, and finally induce the change of MC due to the deposition of brown pigments, i.e. the macrophage phagocytized apoptotic body, on the colonic membrane.
Assuntos
Antraquinonas/efeitos adversos , Catárticos/efeitos adversos , Doenças do Colo/induzido quimicamente , Doenças do Colo/patologia , Melanose/induzido quimicamente , Melanose/patologia , Animais , Apoptose/efeitos dos fármacos , Colo/patologia , Colo/ultraestrutura , Doenças do Colo/sangue , Doenças do Colo/complicações , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/patologia , Regulação da Expressão Gênica/efeitos dos fármacos , Cobaias , Marcação In Situ das Extremidades Cortadas , Masculino , Melanose/sangue , Melanose/complicações , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Fator de Necrose Tumoral alfa/sangue , Fator de Necrose Tumoral alfa/genéticaRESUMO
OBJECTIVE: The aim of this study was to construct a recombinant plasmid carrying fusion suicide gene CDglyTK and RNA interference eukaryotic expressing vector targeting to STAT3, and to investigate the effect of double suicide gene combined with RNAi targeting to STAT3 on HCT116 and HUVEC cells in vitro. METHODS: The CD and TK were cloned by polymerase chain reaction (PCR), and fusion gene CDglyTK was inserted into plasmid pEGFP after DNA sequence analysis, enzyme digestion and ligation. The recombinant plasmid was analyzed by PCR amplification and electrophoresis and enzyme digestion. DNA sequences containing small hairpin structure targeting to STAT3 were synthesized and inserted into the vector. The CDglyTK gene expressions in HCT116 and HUVEC cells were examined by reverse transcription-polymerase chain reaction (RT-PCR) after transfection of HCT116 and HUVEC cells. The inhibitory effect of RNA interference vector targeting to STAT3 was analyzed by RT-PCR and Western blot. The effects of 5-FC and GCV on HCT116 and HUVEC cells transfected with the recombinant plasmids were detected by MTT staining. RESULTS: The results of restriction enzyme digestion and PCR amplification and electrophoresis showed that the recombinant pEGFP/CDglyTK was constructed correctly. The mRNA expression of gene CDglyTK was detected in HCT116 and HUVEC cells which transfected with the recombinant plasmid. The results of RT-PCR and Western blot showed that the RNA interference expression vector targeting to STAT3 effectively inhibited the expression of STAT3 in HCT116 cells. The results of MTT test showed that the inhibition ratio of group pEGFP/CDglyTK was (63.72 ± 0.64)%, significantly higher than that of control group (P < 0.05). The inhibition rate of group pEGFP/STAT3 siRNA was (47.02 ± 0.39)%, which was lower than that of group pEGFP/CDglyTK (P < 0.05), and higher than that of control group (P < 0.05). The inhibition rate of group pEGFP/CdglyTK + pEGFP/STAT3 siRNA was (85.10 ± 0.17)%, significantly higher than those of groups pEGFP/CDglyTK and group pEGFP/STAT3 siRNA (P < 0.05). Meanwhile, in HUVEC cells, the inhibition rate of group pEGFP/CDglyTK was (70.24 ± 0.33)%, significantly higher than that of the control group (P < 0.05). The inhibition rate of group pEGFP/STAT3 siRNA was (46.32 ± 0.15)%, significantly lower than that of group pEGFP/CdglyTK (P < 0.05), and higher than that of the control group (P < 0.05). The inhibition rate of group pEGFP/CdglyTK+pEGFP/STAT3 siRNA was (87.10 ± 0.24)%, significantly higher than those of groups pEGFP/CDglyTK and pEGFP/STAT3 siRNA(P < 0.05). CONCLUSION: The recombinant plasmids pEGFP-CDglyTK and pEGFP/STAT3 siRNA have inhibitory effect on HCT116 and HUVEC cells. The killing effects of double suicide gene combined with RNAi targeting to STAT3 are much better than those of single gene therapy.
Assuntos
Neoplasias Colorretais/terapia , Terapia Genética , Interferência de RNA , Fator de Transcrição STAT3/genética , Linhagem Celular Tumoral , Neoplasias Colorretais/genética , Genes Transgênicos Suicidas , Vetores Genéticos , Humanos , Técnicas In Vitro , Plasmídeos , RNA Interferente Pequeno/genética , TransfecçãoAssuntos
Neoplasias Encefálicas/patologia , Ganglioglioma/patologia , Adulto , Neoplasias Encefálicas/metabolismo , Neoplasias Encefálicas/cirurgia , Feminino , Ganglioglioma/metabolismo , Ganglioglioma/cirurgia , Proteína Glial Fibrilar Ácida/metabolismo , Humanos , Imageamento por Ressonância Magnética , Fosfopiruvato Hidratase/metabolismo , Sinaptofisina/metabolismoRESUMO
OBJECTIVE: To study the induced change of tumor necrosis factor alpha (TNF-alpha) level in serum and colon tissue of guinea pig model with melanosis coli and its significance induced by rhubarb (RB). METHODS: One hundred and twelve guinea pigs of clean grade were randomly divided into four groups: the 16 in the normal group (untreated) and the 3 RB groups (32 in each) treated with low (3 g/kg d), medium (6 g/kg d), and high (12 g/kg d) dose of rhubarb respectively, administered by gastrogavage for 60 successive days. All guinea pigs were sacrificed at the terminal of the experiment and their blood serum and colon tissue were taken for detecting TNF-alpha level and TNF-alpha mRNA expression qualitatively and quantitatively using ELISA and RT-PCR. RESULTS: Compared with the normal group, serum and colonic tissue levels of TNF-alpha and TNF-alpha mRNA expression in the RB groups were higher significantly (P<0.01), while no significant difference was found among the later three groups (P>0.05). CONCLUSION: RB could induce change of TNF-alpha level in serum and colon tissue of guinea pig with melanosis coli.
Assuntos
Doenças do Colo/metabolismo , Medicamentos de Ervas Chinesas/farmacologia , Melanose/metabolismo , Rheum/química , Fator de Necrose Tumoral alfa/metabolismo , Animais , Feminino , Cobaias , Mucosa Intestinal/metabolismo , Masculino , Distribuição AleatóriaRESUMO
Lipopolysaccharide (LPS) is implicated in the pathology of acute liver injury and can induce lethal liver failure when simultaneously administered with D-galactosamine (D-GalN). At the present time, nonlethal liver failure, the liver injury of clinical implication, is incompletely understood following challenge by low-dose LPS/D-GalN. We report here our investigation of the effects of liver injury following a nonlethal dose LPS/D-GalN and the role of apoptosis in this disorder. Blood biochemistry indexes, including those of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and total bilirubin (TBIL), had risen by 6 h post-LPS/D-GalN injection, reached a peak at 24 h and sustained high levels at 48 h. An abnormal liver appearance was found at 24 and 48 h post-injection. Histopathological changes of hepatic injuries accompanied by hepatocellular death, inflammatory infiltration and hemorrhage began to appear at 6 h and were markedly aggravated at 24 and 48 h. Cell apoptosis was significantly induced by the nonlethal dose LPS/D-GalN challenge, and the apoptotic indexes (AIs) in 24 h- and 48 h-treated rats were approximately 70%, as estimated by the terminal transferase dUTP nick end labeling (TUNEL) assay. The mRNA levels of the inflammatory cytokine IL-1beta rose markedly at 6 h and maintained high levels at 24 and 48 h; however, TNF-alpha levels were normal in the liver tissues of 6-, 24- and 48-h-treated rats. mRNA expression of the damage gene nitric oxide synthase (NOS) was also induced early by the LPS/D-GalN challenge, reaching a peak at 6 h, then gradually decreasing in a stepwise manner; conversely, high expression levels of the apoptosis-inducing gene p53 mRNA were not found in the early post-injection period (6 h) but emerged in the crest-time of liver apoptosis (24 h) and were maintained at this level until the late stage (48 h). We also observed that in 24 h-treated rats, caspase-3, -8, -9 and -12 were markedly activated by LPS/D-GalN challenge. These results suggest that a challenge with low-dose LPS in conjunction with D-GalN can induce nonlethal but marked liver failure, the main morphological feature of which is hepatic apoptosis, which may be associated with a high expression of inducible (i)NOS (early post-injection period) and p53 genes (in the mid and late stages) and at least three apoptosis pathways participate in the pathogenesis.
Assuntos
Apoptose/efeitos dos fármacos , Galactosamina/farmacologia , Lipopolissacarídeos/farmacologia , Falência Hepática Aguda/induzido quimicamente , Alanina Transaminase/sangue , Análise de Variância , Animais , Aspartato Aminotransferases/sangue , Bilirrubina/sangue , Caspases/metabolismo , Marcação In Situ das Extremidades Cortadas , Falência Hepática Aguda/patologia , Masculino , Ratos , Ratos Wistar , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
By sol-gel processing, regenerated nano-TiO2/SF (silk fibroin) composite films were synthesized. The experimental results revealed that the nano-TiO2 particles were well dispersed in the regenerated silk fibroin. Using FT-IR and Raman spectroscopy, the secondary structures of these composite films with concentrations of 0, 0.2, 0.4, 0.8 and 1.6 wt% were characterized. Concentration-perturbed two-dimensional (2D) correlation spectra were calculated for the spectra in the 1800-1600 cm(-1) region. To investigate nano-TiO2 particles induced changes in the secondary structure and hydration, the slice spectra were calculated from the synchronous and asynchronous spectra, respectively. The transmittance IR and Raman spectra measurement indicated that the secondary structure of the pure silk film was mostly random coil and alpha-helix, while the composite films were beta-sheet. With increasing nano-TiO2 content, the secondary structure of composite films was changed from typical Silk I to typical Silk II. However, it was found that the transition of the SF's secondary structures would be restrained by excessive nano-TiO2 (over 0.8%) introduced into the composite SF films. Through the FT-IR absorbance and 2D correlation spectra, it was demonstrated that the formation of nano-TiO2 particles could induce the partial transformation of SF conformation from Silk I to Silk II.