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Long-term hypercaloric diets may adversely affect the development of ovarian follicles. We investigated the effects of high sugar (HS), high fat low sugar (HFLS), and high fat normal sugar (HFNS) diets on the ovarian follicle development in mice fed with these diets as compared to those fed with normal diet (control) for 180 days. Body weight, gonadal fat, glucose, lipid, insulin, estrous cycle, sex hormones and ovarian tissues were examined, and metabolism-related protein expression in the ovaries was evaluated by immunoblotting. The mice fed with hypercaloric diets showed hyperinsulinemia and hyperlipidemia, and exhibited heavier body and gonadal fat weights, longer estrous cycles, and fewer preantral and antral follicles than mice fed with normal diet. The sex hormone levels in the blood were similar to those in controls, except for significantly elevated estradiol levels in the HS diet group. The AMPKα phosphorylation was reduced, while AKT phosphorylation and caspase-3 levels were increased in the ovarian tissues of mice in all three hypercaloric diet groups than those in control. Taken together, the results suggest hyperinsulinemia and hyperlipidemia as possible mechanisms that impair the development of ovarian follicles in response to long-term exposure to unhealthy hypercaloric diets.
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Hiperinsulinismo , Hiperlipidemias , Animales , Dieta Alta en Grasa/efectos adversos , Femenino , Glucosa , Hiperinsulinismo/etiología , Hiperlipidemias/etiología , Ratones , Folículo Ovárico/fisiologíaRESUMEN
Lung cancer is a leading cause of cancer-related death worldwide. In this study, we used lung adenocarcinoma cells as a model, as lung adenocarcinoma has the highest mortality rate among all lung cancers. For the past few years, medical treatments or lung cancer have been limited because of chemotherapy resistance. Therefore, understanding the pathogenesis of the development of drug resistance in lung cancer is urgent. Gemcitabine is widely prescribed in the chemotherapeutic treatment of lung cancers. In this study, we developed gemcitabine-resistant lung adenocarcinoma cells (A549-GR) from the A549 cell line. The results showed that apoptotic protein expression and reactive oxygen species (ROS) generation were reduced in A549-GR cells compared to A549 cells. Interestingly, we found that signal transducer and activator of transcription 3 (STAT3) translocated to the nucleus and mitochondria to affect the apoptotic pathway and ROS generation, respectively. Furthermore, treatment with STAT3 small interfering RNA diminished the increase in ROS production, proliferation and antiapoptotic proteins in A549-GR cells. Taken together, the study demonstrated that STAT3 acts as an essential regulator and moderates apoptosis through two major mechanisms to induce gemcitabine resistance in cells; and these findings provide a potential target for the treatment of gemcitabine-resistant lung cancer.
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Apoptosis , Desoxicitidina/análogos & derivados , Resistencia a Antineoplásicos , Neoplasias Pulmonares/metabolismo , Mitocondrias/metabolismo , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Factor de Transcripción STAT3/metabolismo , Células A549 , Apoptosis/efectos de los fármacos , Apoptosis/genética , Núcleo Celular/efectos de los fármacos , Núcleo Celular/metabolismo , Citosol/metabolismo , Desoxicitidina/farmacología , Resistencia a Antineoplásicos/efectos de los fármacos , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Humanos , Neoplasias Pulmonares/patología , MicroARNs/genética , MicroARNs/metabolismo , Mitocondrias/efectos de los fármacos , Modelos Biológicos , Regulación hacia Arriba/efectos de los fármacos , Regulación hacia Arriba/genética , GemcitabinaRESUMEN
Hyperglycemia results in the formation of reactive oxygen species which in turn causes advanced glycation end products (AGEs) formation, leading to diabetic cardiomyopathy. Our previous study showed that AGE-induced reactive oxygen species-dependent apoptosis is mediated via protein kinase C delta (PKCδ)-enhanced mitochondrial damage in cardiomyocytes. By using microRNA (miRNA) database, miRNA-210 was predicted to target c-Jun N-terminal kinase (JNK), which were previously identified as downstream of PKCδ in regulating mitochondrial function. Therefore, we hypothesized that miR-210 mediates PKCδ-dependent upregulation of JNK to cause cardiac mitochondrial damage and apoptosis following AGE exposure. AGE-exposed cells showed activated cardiac JNK, PKCδ, and apoptosis, which were reversed by treatment with a JNK inhibitor and PKCδ-KD (deficient kinase). Cardiac miR-210 and mitochondrial function were downregulated following AGE exposure. Furthermore, JNK was upregulated and involved in AGE-induced mitochondrial damage. Interestingly, luciferase activity of the miR-210 mimic plus JNK WT-3'-untranslated region overexpressed group was significantly lower than that of miR-210 mimic plus JNK MT-3'UTR group, indicating that JNK is a target of miR-210. Moreover, JNK activation induced by AGEs was reduced by treatment with the miR-210 mimic and reversed by treatment with the miR-210 inhibitor, indicating the regulatory function of miR-210 in JNK activation following AGE exposure. Additionally, JNK-dependent mitochondrial dysfunction and apoptosis were reversed following treatment with the miR-210 mimic, while the miR-210 inhibitor showed no effect on JNK-induced mitochondrial dysfunction and apoptosis in AGE-exposed cardiac cells. Taken together, our study showed that PKCδ-enhanced JNK-dependent mitochondrial damage is mediated through the reduction of miR-210 in cardiomyocytes following AGE exposure.
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Apoptosis , Productos Finales de Glicación Avanzada/metabolismo , MAP Quinasa Quinasa 4/metabolismo , MicroARNs/metabolismo , Mitocondrias Cardíacas/metabolismo , Animales , Línea Celular , Productos Finales de Glicación Avanzada/genética , MAP Quinasa Quinasa 4/genética , MicroARNs/genética , Mitocondrias Cardíacas/genética , RatasRESUMEN
Recent studies have provided overwhelming evidence of the involvement of microglia-related molecular networks in the pathogenesis of Alzheimer's diseases (AD). The potential involvement of pro-inflammatory cytokines interleukin (IL)-18, IL-23 and IL-17 on amyloid (Aß) clearance is still unclear. In this study, we addressed that there might be a net relationship among IL-18, IL-23, and IL-17 and they can affect Aß clearance in cultured macrophage/microglia cells. In human macrophage cell line THP-1, Aß42 incubation could increase the expression of IL-18, IL-23 and IL-17 in a concentration dependent manner. THP-1 cell could clear Aß42 in the culture medium time-dependently, but its capacity of Aß clearance was impaired by IL-18, IL-23 or IL-17 treatment. Similarly, the capacity of the microglia cell line BV2 to clear Aß42 was impaired by IL-18, IL-23 or IL-17 treatment. In co-cultures of BV2 with APP/PS1 neuron, Aß was efficiently cleared by BV2 cell, but Aß clearance was impaired by IL-18, IL-23 or IL-17 treatment. The effects of IL-18, IL-23 and IL-17 could be blocked by their corresponding neutralizing antibodies. In addition, the inhibitory effects of IL-18 were blocked by IL-23 or IL-17 neutralizing antibodies while the inhibitory effects of IL-23 were blocked by IL-17 neutralizing antibodies. Our study provides evidences showing that amyloid induced IL-18/IL-23/IL-17 axis could impair macrophage and microglia-mediated Aß clearance. Thus, IL-18/IL-23/IL-17 axis might be a therapeutic target in AD.
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Péptidos beta-Amiloides/metabolismo , Interleucina-17/metabolismo , Interleucina-18/metabolismo , Subunidad p19 de la Interleucina-23/metabolismo , Células THP-1/metabolismo , Enfermedad de Alzheimer/metabolismo , Animales , Encéfalo/metabolismo , Células Cultivadas , Citocinas/metabolismo , Modelos Animales de Enfermedad , Humanos , Ratones , Ratones Transgénicos , Microglía/metabolismo , Neuronas/metabolismoRESUMEN
BACKGROUND: Exosomes are lipid-bilayer enclosed nano-sized vesicles that transfer functional cellular proteins, mRNA and miRNAs. Mesenchymal stem cells (MSCs) derived exosomes have been demonstrated to prevent memory deficits in the animal model of Alzheimer's disease (AD). However, the intravenously injected exosomes could be abundantly tracked in other organs except for the targeted regions in the brain. Here, we proposed the use of central nervous system-specific rabies viral glycoprotein (RVG) peptide to target intravenously-infused exosomes derived from MSCs (MSC-Exo) to the brain of transgenic APP/PS1 mice. MSC-Exo were conjugated with RVG through a DOPE-NHS linker. RESULTS: RVG-tagged MSC-Exo exhibited improved targeting to the cortex and hippocampus after being administered intravenously. Compared with the group administered MSC-Exo, in the group administered RVG-conjugated MSC-Exo (MSC-RVG-Exo) plaque deposition and Aß levels were sharply decreased and activation of astrocytes was obviously reduced. The brain targeted exosomes derived from MSCs was better than unmodified exosomes to improve cognitive function in APP/PS1 mice according to Morris water maze test. Additionally, although MSC-Exo injected intravenously reduced the expression of pro-inflammatory mediators TNF-α, IL-ß, and IL-6, but the changes of anti-inflammatory factors IL-10 and IL-13 were not obvious. However, administration of MSC-RVG-Exo significantly reduced the levels of TNF-α, IL-ß, and IL-6 while significantly raised the levels of IL-10, IL-4 and IL-13. CONCLUSIONS: Taken together, our results demonstrated a novel method for increasing delivery of exosomes for treatment of AD. By targeting exosomes to the cortex and hippocampus of AD mouse, there was a significant improvement in learning and memory capabilities with reduced plaque deposition and Aß levels, and normalized levels of inflammatory cytokines.
RESUMEN
In recent years, neuropathological and epidemiological studies have indicated an association between Alzheimer's disease (AD) and several cardiovascular risk factors. In this study, the cardio-protective effects of folic acid (FA) in early stage AD was elucidated using a triple-transgenic (3xTg) Alzheimer's mouse model. Eleven-month-old C57BL/6 mice and 3xTg mice were assigned to five groups. During the four-month treatment period, the low-FA treatment group received FA through their diet, and the high-FA treatment groups received 3 mg/dl folate in drinking water and were also gastric-fed 1.2 mg/kg folate every day. In the C57B1/6J mice, treatment with high doses of FA (HFA) did not show any considerable effect compared to the control group or the low-dose dietary FA treatment group. However, Alzheimer's mice treated with HFA showed enhanced cardio-protection. Western blot analysis revealed that FA treatment restored SIRT1 expression, which was suppressed in 3xTg mice, through enhanced AMPK expression. FA significantly enhanced the IGF1 receptor survival mechanism in the hearts of the 3xTg mice and suppressed the expression-intrinsic and extrinsic apoptosis-associated proteins. The results suggest that FA intake may significantly alleviate cellular pathological events in the heart associated with AD.
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Enfermedad de Alzheimer/patología , Apoptosis/efectos de los fármacos , Ácido Fólico/farmacología , Enfermedad de Alzheimer/metabolismo , Péptidos beta-Amiloides/genética , Péptidos beta-Amiloides/metabolismo , Animales , Línea Celular , Modelos Animales de Enfermedad , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Miocitos Cardíacos/citología , Miocitos Cardíacos/efectos de los fármacos , Miocitos Cardíacos/metabolismo , Presenilina-1/genética , Presenilina-1/metabolismo , Receptores de Somatomedina/metabolismo , Factores de Riesgo , Sirtuina 1/metabolismo , Superóxido Dismutasa/genética , Superóxido Dismutasa/metabolismo , Proteínas tau/genética , Proteínas tau/metabolismoRESUMEN
Doxorubicin (Dox) causes the generation of intracellular reactive oxygen species (ROS) and inactivates insulin-like growth factor 1 (IGF1) signaling, leading to cardiomyocyte apoptosis. IGF-binding protein 3 (IGFBP3) is the most abundant circulating IGF1 carrier protein with high affinity, which has been reported to mediate ROS-induced apoptosis. Hypoxia-inducible factor 1α (HIF1A), an upstream protein of IGFBP3 is regulated by prolyl hydroxylase domain (PHD) through hydroxylation. In this study, we investigated the role of IGFBP3, HIF1A, and PHD in Dox-induced cardiac apoptosis.Cells challenged with 1 µM Dox for 24 h increased ROS generation, augmented intracellular and secreted IGFBP3 levels, and reduced IGF1 signaling. Further, we showed that Dox enhanced the extracellular association of IGF1 with IGFBP3. Moreover, echocardiography parameters, especially ejection fraction (EF) and fractional shortening (FS) were significantly reduced in ventricle tissue of Dox challenged rats. Notably, siRNA approach against IGFBP3 or an anti-IGFBP3 antibody rescued Dox-induced cardiac apoptosis, mitochondrial ROS, and the decrease in the IGF1 signaling activity. Furthermore, silencing HIF1A either using siRNA or inhibitor downregulated intracellular IGFBP3, rescued apoptosis, mitochondrial generation, and reduction in IGF1 signaling. Finally, western blot data revealed that ROS scavenger reversed Dox-induced cardiac apoptosis, increased levels of HIF1A and secreted IGFBP3, and decreased IGF1 survival signaling and PHD expression.These findings suggest that Dox-induced ROS generation suppressed PHD, which might stabilize nuclear HIF1A protein, leading to increased IGFBP3 expression and secretion. This in turn results in enhanced extracellular association of the latter with IGF1 and blocks IGF1 pro-survival signaling and may result in inducing cardiac apoptosis.
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Doxorrubicina , Proteína 3 de Unión a Factor de Crecimiento Similar a la Insulina , Animales , Ratas , Apoptosis , Doxorrubicina/farmacología , Proteína 3 de Unión a Factor de Crecimiento Similar a la Insulina/genética , Proteína 3 de Unión a Factor de Crecimiento Similar a la Insulina/metabolismo , Miocitos Cardíacos/metabolismo , Especies Reactivas de Oxígeno/metabolismo , ARN Interferente Pequeño/metabolismoRESUMEN
It has been reported that 80% of diabetic patients die due to cardiovascular diseases. We previously demonstrated that activated hypoxia-inducible factor-1α (HIF-1 α)/insulin-like growth factor binding protein-3 (IGFBP-3) signaling by reactive oxygen species (ROS)-regulated prolyl hydroxylase domain-containing protein (PHD) is involved in high-glucose (HG)-induced cardiac apoptosis. Diallyl trisulfide (DATS), a garlic component, shows the strongest inhibitory effect on diabetic cardiomyopathy. In this study, we investigated whether HIF-1α/IGFBP-3 signaling governs the antiapoptotic effect by DATS on HG-exposed cardiomyocytes. It was observed that significantly increased levels of cell apoptosis and decreased Akt phosphorylation were reversed by DATS in HG-exposed cardiac cells. H2O2 and PHD small interfering RNA treatments increased HIF-1α and IGFBP-3 protein levels, which were decreased by DATS treatment. Overexpression of HIF-1α and IGFBP-3 increased HG-induced cell apoptosis, which was suppressed by DATS. The coimmunoprecipitation assay results showed that DATS not only increased the IGF-1 level and reduced IGFBP-3 level but also suppressed their extracellular association for cardiac cells exposed to HG. Experiments using neonatal cardiomyocytes and hearts showed similar results. These findings indicate that the effect of ROS-regulated PHD on the activation of HIF-1α/IGFBP-3 signaling governs the antiapoptotic effect by DATS on HG-exposed cardiomyocytes.
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Proteína 3 de Unión a Factor de Crecimiento Similar a la Insulina , Miocitos Cardíacos , Compuestos Alílicos , Apoptosis , Glucosa , Humanos , Peróxido de Hidrógeno , Subunidad alfa del Factor 1 Inducible por Hipoxia/genética , Proteína 3 de Unión a Factor de Crecimiento Similar a la Insulina/genética , Miocitos Cardíacos/metabolismo , Especies Reactivas de Oxígeno/metabolismo , SulfurosRESUMEN
This study aimed to investigate the mechanism underlying the protective effects of galangin against H2O2/UVB-induced damage using in vitro and in vivo models of photodamage. Moreover, we identified the involvement of miRNA regulation in this process. The H2O2/UVB-treated HS68 human dermal fibroblasts and UVB-induced C57BL/6J nude mice were used as in vitro and in vivo models of photodamage. The results showed that galangin treatment alleviated H2O2/UVB-induced reduction in cell viability, TGFß/Smad signaling impairment, and dermal aging. Based on the results of microRNA array analyses and database searches, hsa-miR-4535 was identified as a potential candidate miRNA that targets Smad4. In vitro, galangin treatment activated Smad2/3/4 complex and inhibited hsa-miR-4535 expression in H2O2/UVB-exposed cells. In vivo, topical application of low (12 mg/kg) and high doses (24 mg/kg) of galangin to the dorsal skin of C57BL/6J nude mice significantly alleviated UVB-induced skin photodamage by promoting TGFß/Smad collagen synthesis signaling, reducing epidermal hyperplasia, wrinkle formation, and skin senescence, as well as inhibiting hsa-miR-4535 expression. Taken together, our findings indicate a link between hsa-miR-4535 and TGFß/Smad collagen synthesis signaling and suggest these factors to be involved in the photo-protective mechanism of galangin in dermal fibroblasts against H2O2/UVB-induced aging. The evidence indicated that galangin with anti-aging properties can be considered as a supplement in skin care products.
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Colágeno/metabolismo , Flavonoides/farmacología , Peróxido de Hidrógeno/efectos adversos , MicroARNs/metabolismo , Protectores contra Radiación/farmacología , Transducción de Señal , Piel/efectos de los fármacos , Proteínas Smad/metabolismo , Factor de Crecimiento Transformador beta/metabolismo , Colágeno/efectos de los fármacos , Colágeno/efectos de la radiación , Fibroblastos/efectos de los fármacos , Fibroblastos/metabolismo , Fibroblastos/efectos de la radiación , Humanos , Transducción de Señal/efectos de los fármacos , Transducción de Señal/efectos de la radiación , Piel/metabolismo , Piel/efectos de la radiación , Envejecimiento de la Piel/efectos de los fármacos , Envejecimiento de la Piel/efectos de la radiación , Rayos Ultravioleta/efectos adversosRESUMEN
To investigate the spatiotemporal properties of epileptiform activity in vitro, 400 microm-thick transverse hippocampal slices were prepared from juvenile rat and planar multi-electrode array (MEA) containing 60 electrodes was used to record the electrical activity induced by bath application of high potassium artificial cerebrospinal fluid (ACSF) on slices. Following successful induction of epileptiform bursts, phenobarbital sodium was applied to test for its inhibitory effects on bursting activity in different regions of slice. Region-specific characteristics of epileptiform activity and anticonvulsant actions of phenobarbital sodium in the hippocampal network were determined by comparing the population activity obtained from MEA. The results showed that: (1) 15 min after high-K+ ACSF application, rhythmic and synchronous epileptiform bursts could be detected from all CA sub-regions. Quantitative analysis indicates that the firing patterns of different CA sub-regions were not statistically different (P>0.05). However, no bursting activity was recorded from granular cells in dentate gyrus, only sparse spikes were observed, with frequency significantly lower than that in CA regions (P<0.05). (2) The high-K+-induced bursting activity could last for more than 40 min with stable bursting activities. (3) Bath application of 60 micromol/L phenobarbital sodium inhibited the bursting activities on hippocampal slice. Bursting activities in CA3c and CA1 were firstly suppressed. 10 min after the phenobarbital sodium application, strong bursting activities persisted only in some of pyramidal cells in CA3a and CA3b. These results show that MEA could be applied for studying the spatial and temporal properties of epileptiform activity in vitro, as well as the region-specific effects of anti-epileptic drugs.
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Potenciales de Acción/fisiología , Epilepsia/fisiopatología , Hipocampo/fisiopatología , Animales , Electrodos , Electroencefalografía , Fenómenos Electrofisiológicos/fisiología , Técnicas In Vitro , Masculino , Ratas , Ratas Sprague-Dawley , Procesamiento de Señales Asistido por ComputadorRESUMEN
OBJECTIVE: To analyze the genetic and biological characters of a new isolate of coxsackievirus B3 (CoxB3), i.e. FY-19 strain, and investigate its mechanistic role in causing different clinical symptoms of hand-foot-mouth disease (HFMD). METHODS: FY-19 strain, isolated from a patient with severe clinical symptoms from Fuyang, China in 2008, was identified by the serological parameters via the Lim Benyesh-Melnick (LBM) antiserum pools. Its genotype was further characterized by sequencing the whole genome. And its biological characters were also examined by proliferation kinetic and pathogenetic analysis. RESULTS: FY-19 strain was identified as CoxB3 showing 23.0%, 16.5% and 32.1% difference with Nancy strain in 3'-, 5'-noncoding and coding regions respectively. FY-19 also showed a high homology with other HFMD-related CoxB3 isolates in China. But its homology with non-HFMD-related CoxB3 isolates was lower (13.5% and 25.0% difference in 3'-NCR and coding region respectively). The viral replication kinetic analysis suggested that the FY-19 proliferation increased rapidly and peaked at 14 hours post-infection. In pathological analysis, FY-19 strain induced mortal pathology in sucking mice. CONCLUSION: Differences in genetic and biological characters exist between FY-19 and Nancy strains. Further analysis on the pathogenesis of this variant may aid in elucidating the mechanisms of HFMD.
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Enterovirus Humano B/clasificación , Enterovirus Humano B/genética , Enfermedad de Boca, Mano y Pie/virología , Animales , Línea Celular , Chlorocebus aethiops , Infecciones por Coxsackievirus , Enterovirus Humano B/aislamiento & purificación , Genotipo , Humanos , Ratones , ARN Viral , Células Vero , Proteínas Virales/genéticaRESUMEN
Local resection or ablation remains an important approach to treat drug-resistant central neurological disease. Conventional surgical approaches are designed to resect the diseased tissues. The emergence of photothermal therapy (PTT) offers a minimally invasive alternative. However, their poor penetration and potential off-target effect limit their clinical application. Here, polydopamine nanoparticles (PDA-NPs) were prepared and characterized. Studies were performed to evaluate whether PDA-NPs combined with near-infrared (NIR) light can be used to ablate deep brain structures in vitro and in vivo. PDA-NPs were prepared with a mean diameter of â¼150 nm. The particles show excellent photothermal conversion efficiency. PDA-NPs did not show remarkable cytotoxicity against neuronal-like SH-SY5Y cell lines. However, it can cause significant cell death when combined with NIR irradiation. Transcranial NIR irradiation after PDA-NPs administration induced enhanced local hyperthermia as compared with NIR alone. Local temperature exceeded 60 °C after 6 min of irradiation plus PDA while it can only reach 48 °C with NIR alone. PTT with PDA (10 mg/mL, 3 µL) and NIR (1.5 W/cm2) can ablate deep brain structures precisely with an ablation volume of â¼6.5 mm3. Histological analysis confirmed necrosis and apoptosis in the targeted area. These results demonstrate the potential of NP-assisted PTT for the treatment against nontumorous central neurological diseases.
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Nanopartículas , Fototerapia , Encéfalo/cirugía , Indoles , PolímerosRESUMEN
OBJECTIVE: To report the experience of surgical resection of Bismuth-Corlette type I and II hilar cholangiocarcinoma. METHODS: From January 1998 and January 2008, 52 cases of Bismuth-Corlette type I and II hilar cholangiocarcinoma were operated on. The clinical data and long-term outcome of the patients was retrospectively analyzed. RESULTS: Of the 52 cases, 44 cases (84.6%) received operation, 28 patients underwent radical resection (63.6%) and 16 patients (36.4%) underwent palliative resection.Seven patients were resected on caudate lobe and other section and lobe of the liver; among them, 2 patients received combined portal vein resection and 4 underwent combined hepatic artery resection respectively. Eleven cases developed postoperative complications and another one died in hospital. The median survival was 33.2 months in radical resection group, and 1-, 3-, 5-year survival rate was 82.6%, 47.8%, 34.7%, respectively, which was significant greater than those in the palliative resection group (41.6%, 16.6%, 8.3%, respectively) (P < 0.05). The median survival was 16.7 months in the palliative resection group. CONCLUSIONS: The radical resection is still the best treatment for Bismuth-Corlette type I and II hilar cholangiocarcinoma. Intraoperative pathology for resection margin, and combined liver resection, portal vein resection and hepatic artery resection can help improve the radical resection rate.
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Neoplasias de los Conductos Biliares/cirugía , Conductos Biliares Intrahepáticos , Colangiocarcinoma/cirugía , Anciano , Anciano de 80 o más Años , Femenino , Estudios de Seguimiento , Hepatectomía , Arteria Hepática/cirugía , Humanos , Masculino , Persona de Mediana Edad , Vena Porta/cirugía , Pronóstico , Estudios Retrospectivos , Tasa de SupervivenciaRESUMEN
BACKGROUND: Transient receptor potential canonical (TRPC) 6 inhibits Aß in Alzheimer's disease (AD) mouse brain and improves the behavioral performance. AIMS: To evaluate the association of TRPC6 expression in peripheral leucocytes from AD and mild cognitive impairment (MCI) patients and to explore its potential value in early diagnosis of AD. METHODS: TRPC6 mRNA levels in peripheral leucocytes were detected by quantitative real-time PCR. The Spearman correlation test was used to ascertain the associations between TRPC6 and the scores of MMSE, ADL, CSDD, CDR. The Receiver Operating Characteristic (ROC) curve was drawn to evaluate the diagnostic potential of TRPC6 for AD and MCI. RESULTS: There were 108â¯CE, 136 MCI, 164 Con and 60 PD in the study. The expression of TRPC6 mRNA level in peripheral leucocytes was significantly lower: 1) in patients with AD and MCI compared to Con; 2) in AD compared to MCI; 3) in hospitalized AD compared to AD from communities. There was a significantly positive correlation between TRPC6 mRNA and MMSE score (pâ¯=â¯.001, Râ¯=â¯0.327). Significantly inverse correlations were found between TRPC6 and CDR score (pâ¯<â¯0.001, Râ¯=â¯-0.303) as well as between TRPC6 and ADL score (pâ¯=â¯.001, Râ¯=â¯-0.342) for all AD. The area under curve of ROC was 0.881 for the classification of AD, and 0.706 for the classification of MCI, respectively. CONCLUSION: TRPC6 expression is inversely correlated with cognitive performance of AD. TRPC6 in peripheral leucocytes may be a potential biomarker for the diagnosis of AD.
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Enfermedad de Alzheimer/metabolismo , Disfunción Cognitiva/metabolismo , Canal Catiónico TRPC6/biosíntesis , Anciano , Estudios de Casos y Controles , Diagnóstico Precoz , Femenino , Hospitalización , Humanos , Leucocitos/metabolismo , Masculino , Persona de Mediana Edad , Enfermedad de Parkinson/metabolismo , Escalas de Valoración Psiquiátrica/estadística & datos numéricosRESUMEN
BACKGROUND: P-glycoprotein (P-gp) mediated drug efflux across the blood-brain barrier (BBB) is an important mechanism underlying poor brain penetration of certain antiepileptic drugs (AEDs). Nanomaterials, as drug carriers, can overcome P-gp activity and improve the targeted delivery of AEDs. However, their applications in the delivery of AEDs have not been adequately investigated. The objective of this study was to develop a nano-scale delivery system to improve the solubility and brain penetration of the antiepileptic drug lamotrigine (LTG). METHODS: LTG-loaded Pluronic(®) P123 (P123) polymeric micelles (P123/LTG) were prepared by thin-film hydration, and brain penetration capability of the nanocarrier was evaluated. RESULTS: The mean encapsulating efficiency for the optimized formulation was 98.07%; drug-loading was 5.63%, and particle size was 18.73 nm. The solubility of LTG in P123/LTG can increase to 2.17 mg/mL, making it available as a solution. The in vitro release of LTG from P123LTG presented a sustained-release property. Compared with free LTG, the LTG-incorporated micelles accumulated more in the brain at 0.5, 1, and 4 hours after intravenous administration in rats. Pretreatment with systemic verapamil increased the rapid brain penetration of free LTG but not P123/LTG. Incorporating another P-gp substrate (Rhodamine 123) into P123 micelles also showed higher efficiency in penetrating the BBB in vitro and in vivo. CONCLUSION: These results indicated that P123 micelles have the potential to overcome the activity of P-gp expressed on the BBB and therefore show potential for the targeted delivery of AEDs. Future studies are necessary to further evaluate the appropriateness of the nanocarrier to enhance the efficacy of AEDs.
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Encéfalo/metabolismo , Portadores de Fármacos/farmacocinética , Nanopartículas/química , Poloxaleno/farmacocinética , Triazinas/farmacocinética , Animales , Química Encefálica , Línea Celular , Portadores de Fármacos/química , Lamotrigina , Masculino , Micelas , Tamaño de la Partícula , Poloxaleno/química , Ratas , Ratas Sprague-Dawley , Triazinas/químicaRESUMEN
To characterize the complete genome sequence of coxsackievirus B1 (CVB1) MSH/KM9/2009 strain isolated from Yunnan, China,2009. Eight overlapping clones covering the whole viral genome (excluding the poly-A tail) were obtained by RT-PCR and sequenced, and their nucleotide and amino acid sequences were compared with other known CVB1 strains. The genome of the CVB1 MSH/KM9/2009 strain had 7384 nucleotides in length, and contained a 741nt non-translated region (NTR) at the 5' end and a 94nt NTR at the 3' end. The entire open reading frame contained 6 549 nt, encoding a 2 183-aa polyprotein. In the coding region, there was no nucleotide deletion or insertion, but some changes of amino acid were unique. The complete genome sequence alignments showed that the CVB1 isolate MSH/KM9/2009 strain shared the highest nucleotide (80.9%, 81.6%, 80.5% and 80%) and amino acid (95.6%, 95.8%, 96.2% and 95.6%) identities to the CVB1 M16560, pmMC, Tucson B1 and CVB1Nm strain, respectively. Phylogenetic tree analysis showed that the MSH/KM9/2009, CVB1 M16560, pmMC, Tucson B1 and CVB1Nm strain clustered into same group. The newly isolated CVB1 strain MSH/KM9/2009 from Yunnan Province belonged to genotype CVB1.
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Infecciones por Coxsackievirus/virología , Enterovirus/genética , Enterovirus/aislamiento & purificación , Genoma Viral , Secuencia de Bases , Preescolar , China , Enterovirus/clasificación , Femenino , Genotipo , Humanos , Lactante , Masculino , Sistemas de Lectura Abierta , Filogenia , Proteínas Virales/genéticaRESUMEN
The epileptic seizure is a dynamic process involving a rapid transition from normal activity to a state of hypersynchronous neuronal discharges. Here we investigated the network properties of epileptiform discharges in hippocampal slices in the presence of high K(+) concentration (8.5 mmol/L) in the bath, and the effects of the anti-epileptic drug valproate (VPA) on epileptiform discharges, using a microelectrode array. We demonstrated that epileptiform discharges were predominantly initiated from the stratum pyramidale layer of CA3a-b and propagated bi-directionally to CA1 and CA3c. Disconnection of CA3 from CA1 abolished the discharges in CA1 without disrupting the initiation of discharges in CA3. Further pharmacological experiments showed that VPA at a clinically relevant concentration (100 µmol/L) suppressed the propagation speed but not the rate or duration of high-K(+)-induced discharges. Our findings suggest that pacemakers exist in the CA3a-b region for the generation of epileptiform discharges in the hippocampus. VPA reduces the conduction of such discharges in the network by reducing the propagation speed.
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Anticonvulsivantes/farmacología , Epilepsia/prevención & control , Hipocampo/efectos de los fármacos , Potasio , Ácido Valproico/farmacología , Animales , Mapeo Encefálico , Región CA3 Hipocampal/efectos de los fármacos , Región CA3 Hipocampal/fisiopatología , Modelos Animales de Enfermedad , Epilepsia/inducido químicamente , Epilepsia/fisiopatología , Hipocampo/fisiopatología , Técnicas In Vitro , Masculino , Células Piramidales/efectos de los fármacos , Células Piramidales/fisiopatología , Ratas , Ratas Sprague-DawleyRESUMEN
OBJECTIVE: To analyze the genetic characterization of the complete genome from a human coxsackievirus B3 strain A103/KM/09 isolated in Yunnan province, 2009. METHODS: By using RT-PCR, all the eight fragments which containing about 1000 nucleotides and covering full viral genome, were sequenced. By using Mega 5.05,Geneious, RDP 3 and SimPlot 3.5.1 software, sequences were aligned with other enterovirus reference sequences. Phylogenetic and recombination analysis were also carried out. RESULTS: The A103/KM/09 isolate genome showed 7389 nucleotides in length , encoding for 2185 amino acids. In the complete genome, the homology of nucleotide and amino acid among the seven coxsackievirus B3 isolates were 81.0%-88.0% and 95.7%-98.0%, respectively. There appeared 81.0% and 95.7% homology when compared with that of Nancy prototype strain. Results from the Phylogenetic analysis showed that the coxsackievirus B3 formed five distinct clades, I-V. Nucleotide divergence rates between clades were 16.2%-24.3% . The A103/KM/09 strain belonged to clade V. Clade V was further divided into four sub-clades,A-D. The nucleotide divergence between sub-clades was 4.3%-11.4%. Putative recombinant event for A103/ KM/09 was detected. CONCLUSION: All coxsackievirus B3 isolates could be divided into five clades, with A103/KM/09 strain belonged to Clade V-D. Evolution of coxsackievirus B3 had occurred in China.
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Encefalitis Viral/virología , Enterovirus Humano B/genética , Enterovirus Humano B/aislamiento & purificación , Infecciones por Enterovirus/virología , Secuencia de Bases , Preescolar , China/epidemiología , Encefalitis Viral/epidemiología , Infecciones por Enterovirus/epidemiología , Genoma Viral , Humanos , Masculino , Filogenia , Proteínas Virales/genéticaRESUMEN
OBJECTIVE: To analyze the genetic characterization of the complete genome from a human echovirus 6 (Echo6) strain KM57-09 isolated in Yunnan, China, in 2009. METHODS: Using the RT-PCR, eight fragments containing about 1000 nucleotides which covered the whole viral genome were sequenced. The sequences were aligned with other reference enterovirus sequences downloaded from the GenBank, using Mega 5.05, RDP 3 and SimPlot 3.5.1 softwares. RESULTS: Similar to the other human enterovirus, KM57-09 isolate genome appeared to have 7419 nucleotides in length, encoding for 2191 amino acids. In the complete genome, the rates of homology on nucleotide and amino acid among the seven Echo6 isolates were 79.3% - 80.2% and 93.3% - 94.4%, respectively as well as 79.3% and 93.6% of the rates of homology when compared with that of D' Amor prototype strain. In different segment of genome. The 2C-3A genome region was most similar to the HN-2-E25 strain, the 5' UTR, VP4, 3D and 3' UTR genome region were most similar to the CoxB5-Henan-2010. In the VP1 gene, the rates of homology on nucleotide and amino acid among the China isolates were 80.0% - 96.0% and 95.8% - 99.0%, respectively, and showed 77.6% - 96.0% and 95.2% - 99.0% of the rates on homology when compared to the other Echo6 reference strains isolated from other countries or areas, respectively. RESULTS: from phylogenetic analysis showed that the Echo6 formed five distinct groups, A-E. The KM57-09 strain belonged to clade E. The nucleotide divergence between clades was 15.6% - 23.3%. The putative recombinant event for KM57-09 was detected with RDP 3, SimPlot 3.5.1 and 3D sequence phylogenetic analysis. CONCLUSION: All the Echo6 isolates could be divided into five clades, the KM57-09 strain belonged to Clade E. The Echo6 strains isolated in China were contributed to several different chains of transmission.
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Echovirus 6 Humano/genética , Echovirus 6 Humano/aislamiento & purificación , Genoma Viral , China , Echovirus 6 Humano/clasificación , Humanos , Filogenia , Homología de Secuencia de AminoácidoRESUMEN
OBJECTIVE: To evaluate and describe computed tomographic (CT) and endoscopic (ES) imaging findings in patients with pathologically confirmed upper gastrointestinal (GI) tract heterotopic pancreas (HP). METHODS: Findings from imaging examinations in 11 patients with pathologically confirmed HP were retrospectively reviewed (CT images obtained from 11 patients and ES images from 6 patients were available for review). Two radiologists evaluated lesion location, size, shape and border as well as growth pattern, enhancement pattern, enhancement grade and number of tumors. The presence of surface dimpling, prominent enhancement of overlying mucosa, and low intralesional attenuation were also evaluated. RESULTS: HP in the upper GI tract showed typical features in CT imaging: submucosal masses, ill-defined borders, endoluminal growth patterns, bright enhancement similar to the normal pancreas, surface dimpling and low intralesional attenuation. Endoscopic photographs manifested an endoluminal, ill-defined, submucosal mass in the upper GI tract wall, typically with central umbilication. The LD (long diameter)/SD (short diameter) ratios were found to be significantly different between HP in the stomach and HP in the duodenum (P<.05 for each finding). In addition, HP in the duodenum tended to be small and round. CONCLUSIONS: HP exhibits typical pancreatic pathologic features. Images showed characteristic features in CT imaging: submucosal masses, ill-defined lesions with an endoluminal growth pattern, bright enhancement similar to the normal pancreas, surface dimpling and low intralesional attenuation. ES imaging showed an endoluminal, ill-defined, submucosal mass, typically with central umbilication.