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Aging results in progressive decline of renal function as well as histological alterations including glomerulosclerosis and interstitial fibrosis. The objective of current study was to test the benefits of moderate swimming exercise in aged rats on renal function and structure and investigate its molecular mechanisms. Aged rats of 21-months old were given moderate swimming exercise for 12 weeks. Swimming exercise in aged rats led to reduced plasma levels of creatinine and blood urea nitrogen. Periodic acid-Schiff staining results revealed reduced renal injury scores in aged rats after swimming exercise. Swimming exercise in aged rats mitigated renal fibrosis and downregulated the mRNA expression of Acta2, Fn, Col1a, Col4a, and Tgfb1 in kidneys. Swimming exercise in aged rats attenuated lipid accumulation and reduced levels of triglyceride in kidneys. Swimming exercise in aged rats abated oxidative stress, evidenced by reduced MDA levels and increased MnSOD activities in kidneys. Swimming exercise in aged rats inhibited NF-κB activities and reduced renal expression of pro-inflammatory cytokines including MCP-1, IL-1ß and IL-6. Mechanistically, swimming exercise restored mRNA and protein expression of PPAR-α in kidney of aged rats. Furthermore, swimming exercise in aged rats increased expression of PPAR-α-targeting microRNAs including miR-21 and miR-34a. Collectively, swimming exercise activated PPAR-α, which partly explained the benefits of moderate swimming exercise in aging kidneys.
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Nefropatias , MicroRNAs , Ratos , Animais , PPAR alfa/metabolismo , Natação , Nefropatias/metabolismo , Rim/metabolismo , Fibrose , MicroRNAs/metabolismo , RNA Mensageiro/metabolismoRESUMO
The Xinjiang brown cattle, Red steppe cattle, and Yunling cattle are indigenous cultivated cattle breeds in Chinese frontier provinces, and they produce high-grade beef and milk products, however, their genetic diversity in many important genes related to excellent meat and milk production is still unknown. Our previous studies have found that several candidate genes (e.g., SREBP1c and PAX7) were associated with bovine economically important phenotypic traits, but none has been reported in the above-mentioned three cattle breeds. Since the InDel (insertion/deletion) marker becomes a useful tool applied in the animal molecular breeding, herein, we firstly found that the InDel variations of seven candidate genes in these cattle. Results showed that the genotypic and allelic distributions of these seven genes were remarkably different among these three cattle (p < 0.05 or p < 0.01). Furthermore, the InDel variations of SREBP1c and PAX7 genes were significantly associated with eight phenotypic traits in Xinjiang brown cattle (p < 0.05 or p < 0.01), respectively, suggesting that they can become the useful DNA markers.
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Bovinos/genética , Mutação INDEL/genética , Fenótipo , Animais , Frequência do Gene/genética , Genótipo , Leite , Fator de Transcrição PAX7/genética , Proteína de Ligação a Elemento Regulador de Esterol 1/genéticaRESUMO
Molybdenum disulfide has drawn persistent interest as a promising nonprecious electrocatalyst alternative to Pt for the hydrogen evolution reaction (HER). However, the MoS2 catalytic efficiency is still lower than the Pt-based catalysts owing to insufficient active sites with more inert basal planes. Herein, we designed and synthesized porous MoS2 nanosheets to activate the basal planes by etching away Al in Al-doped MoS2 . The optimized porous MoS2 shows a small onset overpotential as low as 136â mV, a large cathode current density of 10â mA cm-2 at η=201â mV, a low Tafel slope of 62â mV decade-1 , and a high TOF of 0.29 H2 â s-1 per active site at η=200â mV. This study opens up new avenues for designing electrocatalysts based on porous MoS2 or other layered materials with enhanced HER performance.
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BACKGROUND Chronic hepatitis B virus (HBV) infection is the major cause of hepatocellular carcinoma (HCC). HBV X protein (HBx) plays a crucial role in the development of HCC. Moreover, many tripartite motif (TRIM) family proteins exert diverse biological functions in hepatocarcinogenesis. However, as a novel member of this family, the specific effect of TRIM52 is still largely obscure. In the present study, we investigated the expression and function of TRIM52 in HBV-associated HCC. MATERIAL AND METHODS Fluorescence quantitative polymerase chain reaction (FQ-PCR) was performed to detect the HBV DNA levels in the peripheral blood of HCC patients. Quantitative real-time PCR (qRT-PCR) and Western blot analysis were performed to detect the expression of TRIM52, HBx, and NF-κB p65. HBx-pcDNA3.1 and TRIM52-shRNA were used to induce HBx ectopic expression and TRIM52 silencing, respectively. Pyrrolidine dithiocarbamate (PDTC) was used to block the activation of NF-κB. Cell proliferation was detected using the Cell Counting Kit-8 (CCK-8) assay. RESULTS TRIM52 expression was up-regulated together with HBx in HBV-associated HCC tissues. Ectopic expression of HBx elevated TRIM52 expression in HepG2 cells. TRIM52 silencing repressed the proliferation of HepG2.2.15 cells. Moreover, NF-κB p65 expression was increased in HCC cell lines. Blocking NF-κB activation with PDTC suppressed TRIM52 expression and attenuated the viability of HepG2.2.15 cells. CONCLUSIONS These findings indicate that TRIM52 can promote cell proliferation and HBx may regulate TRIM52 expression via the NF-κB signaling pathway in HBV-associated HCC.
Assuntos
Carcinoma Hepatocelular/metabolismo , Carcinoma Hepatocelular/virologia , Vírus da Hepatite B/fisiologia , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/virologia , Proteínas com Motivo Tripartido/metabolismo , Carcinoma Hepatocelular/patologia , Proliferação de Células , DNA Viral/metabolismo , Regulação Neoplásica da Expressão Gênica , Inativação Gênica , Células Hep G2 , Humanos , Neoplasias Hepáticas/patologia , Transativadores/metabolismo , Fator de Transcrição RelA/metabolismo , Regulação para Cima/genética , Proteínas Virais Reguladoras e AcessóriasRESUMO
Round pneumonia is an uncommon form of pulmonary infection usually found in children. It may resemble pulmonary neoplasm on radiographs. We present a case of round pneumonia in a 43-year-old male with a history of smoking and a family history of lung cancer. The patient was treated with antibiotics for more than two weeks, after which the infection resolved completely both clinically and radiologically. Clinicians should consider this uncommon type of pneumonia in the differential diagnosis of spherical pulmonary masses to avoid unnecessary diagnostic tests.
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Infecções Comunitárias Adquiridas/diagnóstico , Pneumonia/diagnóstico , Adulto , Antibacterianos/uso terapêutico , Infecções Comunitárias Adquiridas/tratamento farmacológico , Diagnóstico Diferencial , Humanos , Neoplasias Pulmonares/diagnóstico , Masculino , Pneumonia/tratamento farmacológico , Radiografia Torácica , Fumar/efeitos adversos , Tomografia Computadorizada por Raios XRESUMO
Tumor necrosis factor-alpha (TNFα) plays a crucial role in inflammatory diseases such as rheumatoid arthritis and postmenopausal osteoporosis. Recently, it has been demonstrated that hydrogen gas, known as a novel antioxidant, can exert therapeutic anti-inflammatory effect in many diseases. In this study, we investigated the effect of treatment with hydrogen molecule (H(2)) on TNFα-induced cell injury in osteoblast. The osteoblasts isolated from neonatal rat calvariae were cultured. It was found that TNFα suppressed cell viability, induced cell apoptosis, suppressed Runx2 mRNA expression, and inhibited alkaline phosphatase activity, which was reversed by co-incubation with H(2). Incubation with TNFα-enhanced intracellular reactive oxygen species (ROS) formation and malondialdehyde production increased NADPH oxidase activity, impaired mitochondrial function marked by increased mitochondrial ROS formation and decreased mitochondrial membrane potential and ATP synthesis, and suppressed activities of antioxidant enzymes including SOD and catalase, which were restored by co-incubation with H(2). Treatment with H(2) inhibited TNFα-induced activation of NFκB pathway. In addition, treatment with H(2) inhibited TNFα-induced nitric oxide (NO) formation through inhibiting iNOS activity. Treatment with H(2) inhibited TNFα-induced IL-6 and ICAM-1 mRNA expression. In conclusion, treatment with H(2) alleviates TNFα-induced cell injury in osteoblast through abating oxidative stress, preserving mitochondrial function, suppressing inflammation, and enhancing NO bioavailability.
Assuntos
Anti-Inflamatórios/farmacologia , Hidrogênio/farmacologia , Osteoblastos/efeitos dos fármacos , Fator de Necrose Tumoral alfa/fisiologia , Fosfatase Alcalina/metabolismo , Animais , Antioxidantes/farmacologia , Apoptose/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Subunidade alfa 1 de Fator de Ligação ao Core/genética , Subunidade alfa 1 de Fator de Ligação ao Core/metabolismo , Inativação Gênica/efeitos dos fármacos , Molécula 1 de Adesão Intercelular/genética , Molécula 1 de Adesão Intercelular/metabolismo , Interleucina-6/genética , Interleucina-6/metabolismo , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , NF-kappa B/metabolismo , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase Tipo II/metabolismo , Osteoblastos/enzimologia , Osteoblastos/imunologia , Ratos , Ratos Sprague-Dawley , Espécies Reativas de Oxigênio/metabolismo , Ativação Transcricional/efeitos dos fármacos , Fator de Necrose Tumoral alfa/farmacologiaRESUMO
BACKGROUND/AIMS: To investigate the suppressive effects of proteasome inhibitor MG132 on hepatitis B virus production. METHODOLOGY: HepG2 2.2.15 hepatoblastoma cells, which constitutively produce HBV particles, were used in the present study. MTT assay was used to evaluate the cytotoxicity of MG132. A Proteasome-Glo chymotrypsin-like cell-based assay was used to access the proteasome activity. Quantitative PCR were performed to analyze HBV-DNA. Secreted HBV antigens in the culture medium were measured by ELISA. Western blot and immunofluorescent staining of HBV antigen were also performed. RESULTS: After 6 days of MG132 treatment, proteasome activity was greatly decreased to 64.3 ± 7.8% and 36.4 ± 7.7% of untreated cells by 0.1µM and 0.3µM of MG132, respectively. The levels of HBsAg and HBeAg, and the copy number of extracellular HBV-DNA, were decreased to nearly half of the control group by 0.1µM MG132. The HBV replicative intermediates were also suppressed by MG132. Western blot and immunofluorescent staining clearly showed the lower levels of the expression of HBV proteins induced by MG132. CONCLUSIONS: MG132 could effectively inhibit the HBV replication in vitro. Ubiquitin-proteasome pathway plays an important role in HBV life cycle and could be a promising therapeutic target for anti-HBV drugs.
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Vírus da Hepatite B/efeitos dos fármacos , Leupeptinas/farmacologia , Inibidores de Proteassoma/farmacologia , Western Blotting , Ensaio de Imunoadsorção Enzimática , Imunofluorescência , Humanos , Técnicas In Vitro , Reação em Cadeia da Polimerase , Sais de Tetrazólio , Tiazóis , Células Tumorais CultivadasRESUMO
In hypertensive animals and patients, oxidative stress represents the primary risk factor for progression of left ventricular hypertrophy. Recently, it has been demonstrated that hydrogen, as a novel antioxidant, can selectively reduce hydroxyl radicals and peroxynitrite anion to exert therapeutic antioxidant activity. In the current study, we explored the effect of chronic treatment with hydrogen-rich saline (HRS) on left ventricular hypertrophy in spontaneously hypertensive rats (SHR). The 8-week-old male SHR and age-matched Wistar-Kyoto rats (WKY) were randomized into HRS-treated (6 ml/kg/day for 3 months, i.p.) and vehicle-treated groups. HRS treatment had no significant effect on blood pressure, but it effectively attenuated left ventricular hypertrophy in SHR. HRS treatment abated oxidative stress, restored the activity of antioxidant enzymes including GPx, GST, catalase, and SOD, suppressed NADPH oxidase activity and downregulated Nox2 and Nox4 expression in left ventricles of SHR. HRS treatment suppressed pro-inflammatory cytokines including IL-1ß, IL-6, TNF-α, and MCP-1, and inhibited NF-κB activation through preventing IκBα degradation in left ventricles of SHR. HRS treatment preserved mitochondrial function through restoring electron transport chain enzyme activity, repressing ROS formation, and enhancing ATP production in left ventricles of SHR. Moreover, HRS treatment suppressed ACE expression and locally reduced angiotensin II generation in left ventricles of SHR. In conclusion, HRS treatment attenuates left ventricular hypertrophy through abating oxidative stress, suppressing inflammatory process, preserving mitochondrial function, in which suppression of HRS on angiotensin II in left ventricles locally might be involved.
Assuntos
Antioxidantes/administração & dosagem , Hidrogênio/administração & dosagem , Hipertensão/complicações , Hipertrofia Ventricular Esquerda/tratamento farmacológico , Estresse Oxidativo , Cloreto de Sódio/administração & dosagem , Angiotensina II/sangue , Angiotensina II/metabolismo , Animais , Catalase/metabolismo , Citocinas/metabolismo , Complexo de Proteínas da Cadeia de Transporte de Elétrons/metabolismo , Glutationa Peroxidase/metabolismo , Glutationa Transferase/metabolismo , Ventrículos do Coração/enzimologia , Ventrículos do Coração/metabolismo , Ventrículos do Coração/patologia , Hemodinâmica , Hipertensão/metabolismo , Hipertrofia Ventricular Esquerda/etiologia , Hipertrofia Ventricular Esquerda/metabolismo , Masculino , Malondialdeído/sangue , Mitocôndrias Cardíacas/enzimologia , Mitocôndrias Cardíacas/fisiologia , NADPH Oxidases/metabolismo , NF-kappa B/metabolismo , Ácido Peroxinitroso/metabolismo , Distribuição Aleatória , Ratos , Ratos Endogâmicos SHR , Ratos Endogâmicos WKY , Espécies Reativas de Oxigênio/metabolismo , Superóxido Dismutase/metabolismoRESUMO
Candida albicans is a common cause of morbidity in hospitalized and immunosuppressed patients. There are still many unknown genes involved in the virulence of C. albicans. The present study aims to examine the effect of TOP2 gene in candidal virulence, including hyphal growth, phospholipase and proteinase activity. Targeted gene disruption of both TOP2 alleles in a wild-type strain of C. albicans produced hyphae more efficiently. TOP2 disruption also increased phospholipase and proteinase activities, and enhanced virulence as assessed by host tissue colonization in systemic infection model. The result of reverse transcription PCR displayed that PLB1 and SAP4 expressions of top2 mutants was significantly upregulated when compared with the isogenic parental strain. Together, these results indicated that TOP2 gene was involved in candidal pathogenicity, and the major reasons for the comparatively high virulence of null mutants were the higher capacity to produce hyphae and the increased phospholipase and proteinase activities, at least in part.
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Candida albicans/genética , Candida albicans/patogenicidade , DNA Topoisomerases Tipo II/genética , Hifas/genética , Animais , Ácido Aspártico Endopeptidases/genética , Ácido Aspártico Endopeptidases/metabolismo , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Regulação Fúngica da Expressão Gênica , Humanos , Hifas/crescimento & desenvolvimento , Camundongos , Camundongos Endogâmicos BALB C , Mutação , Peptídeo Hidrolases/metabolismo , Fosfolipases/metabolismoRESUMO
Cryptococcal osteomyelitis is extremely rare and almost always occurs in immunocompromised patients. We describe a case of osteomyelitis due to Cryptococcus neoformans involving both scapula and rib in an immunocompetent and previously healthy patient. The patient received treatment with amphotericin B deoxycholate and flucytosine for 4 weeks, followed by oral fluconazole 400 mg per day for 8 weeks and 200 mg per day for 9 months. The 12-month course of antifungal therapy resulted in complete clinical recovery and undetectable serum cryptococcal antigen. Cryptococcal osteomyelitis should be suspected in any immunocompetent patient with osteolytic lesions on radiological images.
Assuntos
Criptococose/diagnóstico , Criptococose/microbiologia , Cryptococcus neoformans/isolamento & purificação , Osteomielite/diagnóstico , Osteomielite/microbiologia , Costelas/microbiologia , Escápula/microbiologia , Anfotericina B/administração & dosagem , Antifúngicos/administração & dosagem , Criptococose/patologia , Ácido Desoxicólico/administração & dosagem , Combinação de Medicamentos , Feminino , Flucitosina/administração & dosagem , Humanos , Pessoa de Meia-Idade , Osteomielite/patologia , Costelas/patologia , Escápula/patologia , Fatores de Tempo , Resultado do TratamentoRESUMO
Polycomb group (PcG) proteins are a family of epigenetic regulators responsible for the repression of genes in proliferation and differentiation of stem cells. PcG protein complex consists of two important epigenetic regulators: PRC1 (polycomb repressive complex 1) and PRC2 (polycomb repressive complex 2). In order to further understand the functions of PcG proteins in stem cell growth and differentiation, we review the PcG protein composition, PcG protein localization in the target gene, PcG protein recruitment, and the functions of PcG proteins in the development of stem cells.
Assuntos
Proteínas do Grupo Polycomb/fisiologia , Células-Tronco/citologia , Diferenciação Celular/fisiologia , Proliferação de Células , Humanos , Complexo Repressor Polycomb 1/metabolismo , Complexo Repressor Polycomb 1/fisiologia , Complexo Repressor Polycomb 2/metabolismo , Complexo Repressor Polycomb 2/fisiologia , Proteínas do Grupo Polycomb/metabolismo , Células-Tronco/metabolismoRESUMO
Aging is characterized by inevitable organ function decline over time, with consequent body deterioration and increased susceptibility to death. Astragalus polysaccharide (APS) has been reported to have anti-oxidative, anti-apoptotic, and anti-inflammatory properties. We investigated the potential protective effects of APS on hydrogen peroxide (H2 O2 ) induced hepatocyte senescence and identified related mechanisms in L02, Huh7, and LM3 cell lines. Aged female C57BL/6 mice were given APS for 1 week by intraperitoneal injection, and APS provided the strongest protective effect against H2 O2 -induced damage at 100 µM. APS reduced the expression of cell senescence markers and alleviated pathological damage in aged mouse liver. APS treatment decreased oxidative stress, apoptosis, NOD-like receptor protein-3-mediated pyroptosis, and maintained mitochondrial homeostasis. Notably, the protective effect of APS was weakened in the presence of chloroquine. APS might enrich autophagy by activating AMP-activated protein kinase (AMPK) and inhibiting mammalian target of rapamycin (mTOR). In conclusion, APS reduced reactive oxygen species levels, inhibited apoptosis and pyroptosis, and promoted mitophagy via AMPK/mTOR pathway to alleviate hepatocyte senescence in vitro and in vivo.
Assuntos
Proteínas Quinases Ativadas por AMP , Astrágalo , Proteínas Quinases Ativadas por AMP/genética , Proteínas Quinases Ativadas por AMP/metabolismo , Animais , Apoptose , Astrágalo/metabolismo , Autofagia , Hepatócitos/metabolismo , Mamíferos/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Polissacarídeos/farmacologia , Transdução de Sinais , Serina-Treonina Quinases TOR/genética , Serina-Treonina Quinases TOR/metabolismoRESUMO
The balance of angiotensin-converting enzyme (ACE) and angiotensin-converting enzyme 2 (ACE2) in high blood pressure variability (BPV) induced cardiovascular hypertrophy remains elusive. The aim of the present work was to investigate expression and activity of ACE and ACE2 in the heart and aorta of sinoaortic denervation (SAD) rats with high BPV and normal BP, and explore the potential role of ACE and ACE2 in high BPV-induced cardiovascular damage. Hemodynamics, cardiovascular hypertrophy, angiotensin II (Ang II) concentrations, ACE and ACE2 activity were determined. Cardiac-tissue ACE and ACE2 expression were assayed by real-time polymerase chain reaction and Western blot. Compared with sham-operated rats, systolic BPV and diastolic BPV increased and baroreflex sensitivity decreased significantly in SAD rats. SAD rats presented with obvious cardiovascular hypertrophy characterized by increased ratio of left ventricle weight to body weight and aortic weight to the length of aorta. There was no difference in plasma Ang II concentration between sham-operated and SAD rats. The cardiac and aortic ACE expression, aortic ACE2 expression and ACE activity were elevated in SAD rats. There was no significant difference in cardiac ACE2 expressions between sham-operated and SAD rats. The present work demonstrated that cardiac and aortic ACE expression, aortic ACE2 expression and ACE activity were increased in SAD rats. It is the tissue rather than the circulating renin-angiotensin system that contributes to high BPV-induced cardiovascular hypertrophy.
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Cardiomegalia/enzimologia , Isoenzimas/metabolismo , Peptidil Dipeptidase A/metabolismo , Nó Sinoatrial/inervação , Angiotensina II/sangue , Animais , Barorreflexo , Sequência de Bases , Western Blotting , Primers do DNA , Denervação , Masculino , Ratos , Reação em Cadeia da Polimerase em Tempo RealRESUMO
Important candidate genes that regulate lipid metabolism have the potential to increase the content of intramuscular fat (IMF) and improve meat quality. Secreted protein acidic and rich in cysteine like 1(SPARCL1) is a secreted glycoprotein with important physiological functions and is involved in the proliferation and differentiation of various cells. However, the role of the SPARCL1 gene in sheep preadipocytes and its regulatory mechanism is still unclear. In this study, we explored the effect of SPARCL1 on the proliferation and differentiation of sheep preadipocytes. The results showed that the expression level of the SPARCL1 gene is higher in fat tissue than in other tissues, and the gene was significantly increased on the 6th day of preadipocyte differentiation. In the preadipocyte proliferation stage, interference of SPARCL1 gene reduced cell viability and increased cell apoptosis. In preadipocyte differentiation stage, SPARCL1 overexpression significantly inhibited lipid droplets accumulation and triglyceride content by increasing Wnt10b, Fzd8, IL6, and ß-catenin and inhibiting PPARγ, C/EBPα, LPL, and IGF1 genes expression, whereas SPARCL1 deficiency significantly promoted cell differentiation by inhibiting ß-catenin and increasing GSK3ß, PPARγ, C/EBPα, and LPL. The results of this study suggest that SPARCL1 plays a negative role during preadipocyte differentiation and may become a novel target for regulating preadipocyte differentiation and improving IMF.Abbreviations:IMF: Intramuscular fat SPARCL1: Secreted protein acidic and rich in cysteine like 1 PPARγ: Peroxisome proliferator-activated receptor γ C/EBPα: CCAAT/enhancer-binding protein-α LPL: Lipoprotein lipase IGF1: Insulin-like growth factor 1 Wnt10b: Wnt family member 10B Fzd8: Frizzled class receptor 8 IL6: Interleukin 6 ß-catenin: Catenin beta interacting protein 1 GSK3ß: Glycogen synthase kinase 3 beta LRP5/6: Low-density lipoprotein receptor-related protein 5/6.
Assuntos
Adipócitos , Proteínas de Ligação ao Cálcio/metabolismo , Proliferação de Células , Proteínas da Matriz Extracelular/metabolismo , Adipócitos/citologia , Adipogenia , Animais , Proteína alfa Estimuladora de Ligação a CCAAT/metabolismo , Diferenciação Celular , OvinosRESUMO
The present study was designed to test the hypothesis that a small dose of ketanserin, which enhances baroreflex activity, prevents the early lesions of atherosclerosis. In experiment 1, baroreflex sensitivity (BRS) was measured in 31 spontaneously hypertensive rats (SHRs) in a conscious state using a computerized blood pressure monitoring system. Four weeks later, the rats were administered vitamin D3 and fed a high-cholesterol diet for 8 weeks to induce atherosclerosis. Then their hearts and aortae were removed for pathological examination. A negative correlation was found between BRS and the scores of coronary (r = -0.460, P < 0.01) and aortic atherosclerosis (r = -0.448, P < 0.05) in SHR. In experiment 2, SHRs were divided into 3 groups (n = 10 in each group) and received a dose of ketanserin of 0.3, 1.0, and 3.0 mg/kg (i.g.), respectively. At the smallest dose (0.3 mg/kg), ketanserin did not lower blood pressure but enhanced BRS. In experiment 3, SHRs were administered vitamin D3, fed a high-cholesterol diet, and simultaneously treated with low-dose ketanserin. The atherosclerosis scores of the treatment group were significantly lower than those of the control group (coronary score: 0.90 ± 0.14 vs. 1.76 ± 0.27, P < 0.05; aortic scores: 1.00 ± 0.39 vs. 2.18 ± 0.41, P < 0.05). In experiment 4, male New Zealand White rabbits were fed a high-cholesterol diet and treated with low-dose ketanserin at the same time. The atherosclerosis scores of the treatment group were significantly lower than those of the control group (aortic scores: 0.26 ± 0.20 vs. 0.60 ± 0.31, P < 0.05). In conclusion, the present study demonstrated, for the first time, that low-dose ketanserin prevented the development of atherosclerosis independent of its blood pressure lowering action in SHRs and New Zealand White rabbits at least in part via enhancement of arterial baroreflex function.
Assuntos
Anti-Hipertensivos/uso terapêutico , Aterosclerose/prevenção & controle , Ketanserina/uso terapêutico , Antagonistas da Serotonina/uso terapêutico , Animais , Aterosclerose/fisiopatologia , Barorreflexo/efeitos dos fármacos , Pressão Sanguínea/efeitos dos fármacos , Feminino , Masculino , Coelhos , Ratos , Ratos Endogâmicos SHRRESUMO
BACKGROUND: Homeobox A10 (HOXA10) has been implicated in the development and progression of various human cancers. However, the precise biological functions of HOXA10 in hepatocellular carcinoma (HCC) have not been defined. METHODS: In this study, we examined mRNA expression by quantitative real-time PCR (qRT-PCR) of HOXA10 as well as histone deacetylase (HDAC) and protein levels by Western blot of HOXA10, HDAC1, Cyclin D1, proliferating cell nuclear antigen (PCNA), Survivin and p53 acetylation in HCC tissues and cell lines. We also assessed cell proliferation using Cell Counting Kit-8 (CCK-8) and analyzed cell cycle by flow cytometry. Furthermore, tumor growth of HCC cells in vivo was monitored using the nude mouse xenograft model. Finally, HDAC1 promoter activity and binding in HCC cell lines were detected by luciferase reporter assay and chromatin immunoprecipitation (ChIP), respectively. RESULTS: We uncovered the elevated expression of HOXA10 in HCC tissues compared to adjacent normal liver tissues. RNA interference-mediated knockdown of HOXA10 inhibited HCC cell proliferation both in vitro and in vivo. HOXA10 knockdown also induced cell cycle arrest at G0/G1 phase and apoptosis, which were accompanied with the reduced expression of Cyclin D1, PCNA and Survivin. Notably, HOXA10 knockdown enhanced p53 acetylation (Lys382), which is crucial to the activation of p53. Likewise, HOXA10 knockdown suppressed the transcription of HDAC1, a potential deacetylase for p53. In line with these observations, HDAC1 downregulation abrogated the effects of HOXA10 overexpression on proliferation, cell cycle progression, apoptosis and p53 acetylation, indicating the role of HDAC1 in mediating HOXA10 functions. CONCLUSION: Our results demonstrate that HOXA10 knockdown inhibits proliferation, induces cell cycle arrest and apoptosis in HCC cells by regulating HDAC1 transcription.
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Polyamines are key regulators in cell growth and differentiation. It has been shown that ornithine decarboxylase (Odc) was essential for post-implantation embryo development, and overexpression of spermidine/spermine N1-acetyltransferase will lead to ovarian hypofunction and hypoplastic uteri. However, the expression and function of polyamine-related genes in mouse uterus during early pregnancy are still unknown. In this study we investigated the expression, regulation, and function of polyamine-related genes in mouse uterus during the peri-implantation period. Odc expression was strongly detected at implantation sites and stimulated by estrogen treatment. The expression of Odc antizyme 1 and spermidine/spermine N1-acetyltransferase was also highly shown at implantation sites and regulated by Odc or polyamine level in uterine cells. Embryo implantation was significantly inhibited by alpha-difluoromethylornithine, an Odc inhibitor. Moreover, the reduction of Odc activity caused by alpha-difluoromethylornithine treatment was compensated by the up-regulation of S-adenosylmethionine decarboxylase gene expression. Collectively, our results indicated that the coordinated expression of uterine polyamine-related genes may be important for embryo implantation.
Assuntos
Implantação do Embrião/efeitos dos fármacos , Implantação do Embrião/genética , Regulação da Expressão Gênica/efeitos dos fármacos , Poliaminas/farmacologia , Útero/efeitos dos fármacos , Adenosilmetionina Descarboxilase/genética , Animais , Eflornitina/farmacologia , Feminino , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Hormônios Esteroides Gonadais/farmacologia , Masculino , Camundongos , Ornitina Descarboxilase/genética , Ornitina Descarboxilase/metabolismo , Ornitina Descarboxilase/fisiologia , Ovariectomia , Oxirredutases atuantes sobre Doadores de Grupo CH-NH/genética , Oxirredutases atuantes sobre Doadores de Grupo CH-NH/metabolismo , Poliaminas/metabolismo , Gravidez , Proteínas/genética , Proteínas/metabolismo , Pseudogravidez/genética , Útero/metabolismo , Poliamina OxidaseRESUMO
AIM: To explore the prevalence of SEN virus (SENV) in patients with non A-E hepatitis and volunteer blood donors in Shanghai. METHODS: According to the published gene sequences, primers from the conserved region were designed. Then, the prevalence of SEN virus in 30 samples from healthy voluntary blood donors and 30 samples from patients with non A-E hepatitis were detected by nested-PCR of SENV-D/H. Some PCR products were cloned and sequenced. RESULTS: The specificity of genotype-specific PCR was confirmed by sequencing, the SENV DNA was detected in 53.3% of the patients with non A-E hepatitis and 10% of the blood donors. The prevalence of SENV-D/H viremia was significantly higher in patients with non A-E hepatitis than in blood donors (P = 0.0002). SENV-H subtype and SENV-D subtype were found in 2 and 1 samples, respectively from blood donors. SENV-H subtype, SENV D subtype, mixed SENV-D and SENV-H subtype were found in 8, 6 and 2 samples, respectively, from patients with non A-E hepatitis. CONCLUSION: The gene type of SENV in patients with non A-E hepatitis and blood donors in shanghai is D or H subtype, and transfusion is not the only transmitting form of SENV.
Assuntos
Doadores de Sangue , Hepatite Viral Humana/virologia , Torque teno virus/isolamento & purificação , Adulto , Idoso , Sequência de Bases , DNA Viral/análise , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Dados de Sequência Molecular , PrevalênciaRESUMO
A chemically modified electrode is constructed based on the multi-walled carbon nanotubes (MWNTs)/4-aminobenzeresulfonic acid (4-ABSA) film-coated glassy carbon electrode. The electrocatalytic oxidation of tyrosine (Tyr) is investigated on the surface of the MWNTs/4-ABSA-modified electrode using cyclic voltammetry (CV) and differential pulse voltammetry (DPV). The prepared modified electrode shows voltammetric responses with high sensitivity and selectivity for Tyr in optimal conditions, which makes it very suitable for sub-micromolar detection of Tyr. A sensitive oxidation peak at +0.64 V is employed to determine Tyr. Good linear relationship between the oxidation peak current and the Tyr concentration in the range of 1x10(-7) to 5x10(-5) mol/L is obtained in phosphate buffer solution with pH 7.0. By use of modified electrode, the voltammetric detection limit for Tyr in DPV measurements is 8x10(-8) mol/L (S/N=3). Good sensitivity, selectivity and stability of the low-cost modified electrode make it very suitable for the determination of trace amounts of Tyr in pharmaceutical and clinical preparations.
Assuntos
Eletroquímica/métodos , Vidro/química , Nanotubos de Carbono/química , Ácidos Sulfanílicos/química , Ácidos Sulfônicos/química , Tirosina/análise , Calibragem , Oxirredução , Preparações Farmacêuticas/química , Reprodutibilidade dos Testes , Fatores de TempoRESUMO
limitations in current technology for generating transgenic animals, such as the time and the expense, hampered its extensive use in recombinant protein production for therapeutic purpose. In this report, we present a simple and less expensive alternative by directly infusing a recombinant adenovirus vector carrying human lactoferrin cDNA into rabbit mammary glands. The milk serum was collected from the infected mammary gland 48 h post-infection and subjected to a 10% SDS-PAGE and Western blotting. An 80-kDa protein was visualized after viral vector infection. With this method, we obtained a high level of expressed human lactoferrin of up to 2.3 mg/ml in the milk. Taken together, the method is useful for the transient high-level expression recombinant proteins, and the approach established here is probably one of the most economical and efficient ways for large-scale production of recombinant proteins of biopharmaceutical interest.