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1.
Eur J Nutr ; 61(7): 3697-3706, 2022 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-35689124

RESUMO

PURPOSE: Serum magnesium is the most frequently used laboratory test for evaluating clinical magnesium status. Hypomagnesemia (low magnesium status), which is associated with many chronic diseases, is diagnosed using the serum magnesium reference range. Currently, no international consensus for a magnesemia normal range exists. Two independent groups designated 0.85 mmol/L (2.07 mg/dL; 1.7 mEq/L) as the low cut-off point defining hypomagnesemia. MaGNet discussions revealed differences in serum magnesium reference ranges used by members' hospitals and laboratories, presenting an urgent need for standardization. METHODS: We gathered and compared serum magnesium reference range values from our institutions, hospitals, and colleagues worldwide. RESULTS: Serum magnesium levels designating "hypomagnesemia" differ widely. Of 43 collected values, only 2 met 0.85 mmol/L as the low cut-off point to define hypomagnesemia. The remainder had lower cut-off values, which may underestimate hypomagnesemia diagnosis in hospital, clinical, and research assessments. Current serum magnesium reference ranges stem from "normal" populations, which unknowingly include persons with chronic latent magnesium deficit (CLMD). Serum magnesium levels of patients with CLMD fall within widely used "normal" ranges, but their magnesium status is too low for long-term health. The lower serum magnesium reference (0.85 mmol/L) proposed specifically prevents the inclusion of patients with CLMD. CONCLUSIONS: Widely varying serum magnesium reference ranges render our use of this important medical tool imprecise, minimizing impacts of low magnesium status or hypomagnesemia as a marker of disease risk. To appropriately diagnose, increase awareness of, and manage magnesium status, it is critical to standardize lower reference values for serum magnesium at 0.85 mmol/L (2.07 mg/dL; 1.7 mEq/L).


Assuntos
Magnésio , Humanos , Padrões de Referência , Valores de Referência
2.
BMC Neurosci ; 19(1): 37, 2018 06 25.
Artigo em Inglês | MEDLINE | ID: mdl-29940857

RESUMO

BACKGROUND: High-frequency transcutaneous neuromuscular electrical nerve stimulation (TENS) is currently used for the administration of electrical current in denervated muscle to alleviate muscle atrophy and enhance motor function; however, the time window (i.e. either immediate or delayed) for achieving benefit is still undetermined. In this study, we conducted an intervention of sciatic nerve crush injury using high-frequency TENS at different time points to assess the effect of motor and sensory functional recovery. RESULTS: Animals with left sciatic nerve crush injury received TENS treatment starting immediately after injury or 1 week later at a high frequency(100 Hz) or at a low frequency (2 Hz) as a control. In SFI gait analysis, either immediate or late admission of high-frequency electrical stimulation exerted significant improvement compared to either immediate or late administration of low-frequency electrical stimulation. In an assessment of allodynia, immediate high frequency electrical stimulation caused a significantly decreased pain threshold compared to late high-frequency or low-frequency stimulation at immediate or late time points. Immunohistochemistry staining and western blot analysis of S-100 and NF-200 demonstrated that both immediate and late high frequency electrical stimulation showed a similar effect; however the effect was superior to that achieved with low frequency stimulation. Immediate high frequency electrical stimulation resulted in significant expression of TNF-α and synaptophysin in the dorsal root ganglion, somatosensory cortex, and hippocampus compared to late electrical stimulation, and this trend paralleled the observed effect on somatosensory evoked potential. The CatWalk gait analysis also showed that immediate electrical stimulation led to a significantly high regularity index. In primary dorsal root ganglion cells culture, high-frequency electrical stimulation also exerted a significant increase in expression of TNF-α, synaptophysin, and NGF in accordance with the in vivo results. CONCLUSION: Immediate or late transcutaneous high-frequency electrical stimulation exhibited the potential to stimulate the motor nerve regeneration. However, immediate electrical stimulation had a predilection to develop neuropathic pain. A delay in TENS initiation appears to be a reasonable approach for nerve repair and provides the appropriate time profile for its clinical application.


Assuntos
Lesões por Esmagamento/terapia , Regeneração Nervosa/fisiologia , Neuralgia/fisiopatologia , Nervo Isquiático/lesões , Estimulação Elétrica Nervosa Transcutânea , Animais , Estimulação Elétrica/métodos , Potenciais Somatossensoriais Evocados/fisiologia , Masculino , Ratos Sprague-Dawley , Neuropatia Ciática/metabolismo , Estimulação Elétrica Nervosa Transcutânea/métodos
3.
J Pathol ; 241(3): 337-349, 2017 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-27801527

RESUMO

Hypoxia-inducible factor 1α (HIF-1α) controls many genes involved in physiological and pathological processes. However, its roles in glutamatergic transmission and excitotoxicity are unclear. Here, we proposed that HIF-1α might contribute to glutamate-mediated excitotoxicity during cerebral ischaemia-reperfusion (CIR) and investigated its molecular mechanism. We showed that an HIF-1α conditional knockout mouse displayed an inhibition in CIR-induced elevation of extracellular glutamate and N-methyl-d-aspartate receptor (NMDAR) activation. By gene screening for glutamate transporters in cortical cells, we found that HIF-1α mainly regulates the cystine-glutamate transporter (system xc- ) subunit xCT by directly binding to its promoter; xCT and its function are up-regulated in the ischaemic brains of rodents and humans, and the effects lasted for several days. Genetic deletion of xCT in cortical cells of mice inhibits either oxygen glucose deprivation/reoxygenation (OGDR) or CIR-mediated glutamate excitotoxicity in vitro and in vivo. Pharmaceutical inhibition of system xc- by a clinically approved anti-cancer drug, sorafenib, improves infarct volume and functional outcome in rodents with CIR and its therapeutic window is at least 3 days. Taken together, these findings reveal that HIF-1α plays a role in CIR-induced glutamate excitotoxicity via the long-lasting activation of system xc- -dependent glutamate outflow and suggest that system xc- is a promising therapeutic target with an extended therapeutic window in stroke. Copyright © 2016 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.


Assuntos
Sistema y+ de Transporte de Aminoácidos/metabolismo , Isquemia Encefálica/metabolismo , Encéfalo/metabolismo , Hipóxia Celular/fisiologia , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Sistema y+ de Transporte de Aminoácidos/genética , Animais , Separação Celular/métodos , Ácido Glutâmico/metabolismo , Camundongos , Ativação Transcricional/fisiologia , Regulação para Cima
5.
BMC Musculoskelet Disord ; 18(1): 171, 2017 04 24.
Artigo em Inglês | MEDLINE | ID: mdl-28438150

RESUMO

BACKGROUND: Intermittent parathyroid hormone (PTH) can be used to treat osteoporosis of the spine and hip. However, whether it can be used to treat osteoporosis of the mandible is unclear. The purpose of this study was to explore the influence of applying intermittent PTH to ovariectomized rats on the trabecular bone microarchitecture of the mandible and femoral head. METHODS: Eighteen female rats were divided into three groups: the healthy group, ovariectomized (OVX) group, and OVX + PTH group. The OVX group and OVX + PTH group had an OVX at 8 weeks of age. The OVX + PTH group received intermittent PTH therapy for 12 weeks. The mandibles and femurs of all rats were removed at 20 weeks and were then scanned using microcomputed tomography (micro-CT). RESULTS: From the micro-CT analysis, the trabecular bone microarchitecture of the mandible and femoral head are offered as follows: (1) The bone volume fraction and trabecular thickness in the OVX group were lower than those in the healthy group. (2) The bone volume fraction and trabecular thickness in the OVX + PTH group approximated those in the healthy group. CONCLUSION: The conclusions of this study regarding the trabecular bone microarchitecture of the mandible and femoral head are offered as follows: (1) The BV/TV and TbTh in the OVX group were lower than those in the healthy group. (2) The BV/TV and TbTh in the OVX + PTH group approximated those in the healthy group, therefore, intermittent PTH displayed high efficacy for treating femoral or mandibular deterioration of bone microstructure resulting from loss of ovarian function. Osteoporosis of the femur or mandible in the rats was ameliorated by intermittent PTH therapy.


Assuntos
Cabeça do Fêmur/efeitos dos fármacos , Cabeça do Fêmur/diagnóstico por imagem , Mandíbula/efeitos dos fármacos , Mandíbula/diagnóstico por imagem , Ovariectomia/efeitos adversos , Hormônio Paratireóideo/administração & dosagem , Animais , Feminino , Ovariectomia/tendências , Ratos , Ratos Wistar , Microtomografia por Raio-X/métodos
6.
Environ Toxicol ; 32(2): 456-468, 2017 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-26919256

RESUMO

Cinnamomum cassia essential oil (CC-EO) has various functional properties, such as anti-microbial, hypouricemic, anti-tyrosinase and anti-melanogenesis activities. The present study aimed to evaluate the anti-cancer activities of CC-EO and its major constituent, cinnamaldehyde, in human oral squamous cell carcinoma HSC-3 cells. Determination of the cell viability, apoptotic characteristics, DNA damage, cell cycle analysis, reactive oxygen species (ROS) production, mitochondrial membrane potential, cytosolic Ca2+ level and intracellular redox status were performed. Our results demonstrated that CC-EO and cinnamaldehyde significantly decreased cell viability and caused morphological changes. The cell cycle analysis revealed that CC-EO and cinnamaldehyde induced G2/M cell cycle arrest in HSC-3 cells. The apoptotic characteristics (DNA laddering and chromatin condensation) and DNA damage were observed in the CC-EO-treated and cinnamaldehyde-treated HSC-3 cells. Moreover, CC-EO and cinnamaldehyde promoted an increase in cytosolic Ca2+ levels, induced mitochondrial dysfunction and activated cytochrome c release. The results of ROS production and intracellular redox status demonstrated that CC-EO and cinnamaldehyde significantly increased the ROS production and thiobarbituric acid reactive substance levels, and the cellular glutathione content and glutathione peroxidase activity were significantly reduced in HSC-3 cells. Our results suggest that CC-EO and cinnamaldehyde may possess anti-oral cancer activity in HSC-3 cells. © 2016 Wiley Periodicals, Inc. Environ Toxicol 32: 456-468, 2017.


Assuntos
Acroleína/análogos & derivados , Antineoplásicos Fitogênicos/farmacologia , Apoptose/efeitos dos fármacos , Pontos de Checagem do Ciclo Celular/efeitos dos fármacos , Cinnamomum/química , Óleos Voláteis/farmacologia , Acroleína/farmacologia , Carcinoma de Células Escamosas/tratamento farmacológico , Carcinoma de Células Escamosas/patologia , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Dano ao DNA , Humanos , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Neoplasias Bucais/tratamento farmacológico , Neoplasias Bucais/patologia , Espécies Reativas de Oxigênio/metabolismo
7.
Eur Arch Otorhinolaryngol ; 272(10): 3051-7, 2015 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-25726166

RESUMO

We aimed to investigate expression of magnesium transporter genes in patients with head and neck cancer who underwent cisplatin-based neoadjuvant chemotherapy and their association with serum magnesium level. Head and neck cancer patients scheduled to undergo neoadjuvant cisplatin-based chemotherapy were eligible for enrollment. Blood samples were obtained at three time points: prior to, during, and after completion of chemotherapy. Expression levels of magnesium transporter genes were determined by quantitative real-time PCR. A total of 23 patients were included in the final analysis. The average serum magnesium levels dropped 6.98 and 5.20% during and after completion of chemotherapy. There were neither significant associations between serum magnesium level and demographic variables nor tumor-related variables. SLC41A1 expression level was positively correlated with serum magnesium whereas TRPM6 expression level was negatively correlated with serum magnesium. Serum magnesium level decreased during cisplatin-based chemotherapy in head and neck cancer patients. Further studies are warranted to investigate optimal magnesium measurement and substitution protocol.


Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica , Proteínas de Transporte de Cátions/metabolismo , Cisplatino , Neoplasias de Cabeça e Pescoço , Canais de Cátion TRPM/metabolismo , Desequilíbrio Hidroeletrolítico , Adulto , Idoso , Antineoplásicos/administração & dosagem , Antineoplásicos/efeitos adversos , Protocolos de Quimioterapia Combinada Antineoplásica/efeitos adversos , Protocolos de Quimioterapia Combinada Antineoplásica/farmacologia , Cisplatino/administração & dosagem , Cisplatino/efeitos adversos , Monitoramento de Medicamentos/métodos , Perfilação da Expressão Gênica , Neoplasias de Cabeça e Pescoço/tratamento farmacológico , Neoplasias de Cabeça e Pescoço/patologia , Humanos , Magnésio/sangue , Masculino , Pessoa de Meia-Idade , Terapia Neoadjuvante/efeitos adversos , Terapia Neoadjuvante/métodos , Estadiamento de Neoplasias , Desequilíbrio Hidroeletrolítico/sangue , Desequilíbrio Hidroeletrolítico/induzido quimicamente , Desequilíbrio Hidroeletrolítico/fisiopatologia
8.
Cardiovasc Diabetol ; 13: 120, 2014 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-25078288

RESUMO

BACKGROUND: We investigated the effects of dietary calcium (Ca) and magnesium (Mg) intakes on cardiovascular disease risks in older patients with diabetes. METHODS: In this cross-sectional study, 197 patients with type 2 diabetes aged 65 years and above were recruited. The 24-h dietary recalls and 1-week self-reported typical dietary intake patterns were collected. The Ca and Mg intakes of <67% of the recommended dietary allowance (RDA), 67%-100% of RDA, and >100% of RDA were defined as low, moderate, and high Ca and Mg intakes, respectively. Anthropometric measurements were determined and biochemical analysis of blood and urine was performed. RESULTS: Our data indicated that 60.9% and 87.3% of our patients were Ca and Mg intakes below RDA, respectively. Patients whose Ca intake was high or low (81.2%) had significantly higher C-reactive protein (CRP) than those whose Ca intake was moderate (p = 0.043). Furthermore, patients whose Mg intake was low (87.3%) had significantly higher CRP than that of those who took adequate Mg (p = 0.025). The dietary Ca:Mg intake ratios were highly correlated with CRP, platelet counts, and red blood cell distribution (p < 0.05). A dietary Ca:Mg intake ratio of 2.0-2.5 was significantly correlated to lower CRP levels (p = 0.013). CONCLUSIONS: High or low calcium intake increases cardiovascular disease risks. We suggest that "moderate" intake of 402-600 mg Ca/day (approximately 67%-100% of Taiwan RDA for Ca) and adequate Mg intake (or meeting RDA for Mg) with Ca:Mg intake ratio of 2.0-2.5 are important for reducing cardiovascular disease risks in older patients with diabetes.


Assuntos
Cálcio da Dieta/administração & dosagem , Cálcio da Dieta/efeitos adversos , Doenças Cardiovasculares/epidemiologia , Diabetes Mellitus Tipo 2/epidemiologia , Idoso , Idoso de 80 Anos ou mais , Biomarcadores/sangue , Proteína C-Reativa/metabolismo , Doenças Cardiovasculares/sangue , Doenças Cardiovasculares/induzido quimicamente , Estudos Transversais , Diabetes Mellitus Tipo 2/sangue , Feminino , Humanos , Magnésio/administração & dosagem , Magnésio/efeitos adversos , Masculino , Fatores de Risco
9.
J Neuroeng Rehabil ; 11: 62, 2014 Apr 16.
Artigo em Inglês | MEDLINE | ID: mdl-24739213

RESUMO

BACKGROUND: A new version of the CatWalk XT system was evaluated as a tool for detecting very subtle alteration in gait based on higher speed sample rate; the system could also demonstrate minor changes in neurological function. In this study, we evaluated the neurological outcome of sciatic nerve injury intervened by local injection of hyaluronic acid. Using the CatWalk XT system, we looked for differences between treated and untreated groups and differences within the same group as a function of time so as to assess the power of the Catwalk XT system for detecting subtle neurological change. METHODS: Peripheral nerve injury was induced in 36 Sprague-Dawley rats by crushing the left sciatic nerve using a vessel clamp. The animals were randomized into one of two groups: Group I: crush injury as the control; Group II: crush injury and local application with hyaluronic acid. These animals were subjected to neurobehavior assessment, histomorphology evaluation, and electrophysiology study periodically. These data were retrieved for statistical analysis. RESULTS: The density of neurofilament and S-100 over the distal end of crushed nerve showed significant differences either in inter-group comparison at various time points or intra-group comparison from 7 to 28 days. Neuronal structure architecture, axon counts, intensity of myelination, electrophysiology, and collagen deposition demonstrate significant differences between the two groups. There was significant difference of SFI and angle of ankle in inter- group analysis from 7 to 28 days, but there were no significant differences in SFI and angle of ankle at time points of 7 and 14 days. In the Cat Walk XT analysis, the intensity, print area, stance duration, and swing duration all showed detectable differences at 7, 14, 21, and 28 days, whereas there were no significant difference at 7 and 14 days with CatWalk 7 testing. In addition, there were no significant differences of step sequence or regularity index between the two versions. CONCLUSION: Hyaluronic acid augmented nerve regeneration as early as 7 days after crush injury. This subtle neurological alteration could be detected through the CatWalk XT gait analysis but not the SFI, angle of ankle, or CatWalk 7 methods.


Assuntos
Ácido Hialurônico/farmacologia , Coxeadura Animal/diagnóstico , Traumatismos dos Nervos Periféricos/diagnóstico , Animais , Eletrofisiologia , Imuno-Histoquímica , Coxeadura Animal/etiologia , Compressão Nervosa , Regeneração Nervosa/efeitos dos fármacos , Traumatismos dos Nervos Periféricos/complicações , Traumatismos dos Nervos Periféricos/fisiopatologia , Ratos , Ratos Sprague-Dawley , Recuperação de Função Fisiológica/efeitos dos fármacos , Nervo Isquiático/lesões
10.
Nutr J ; 11: 41, 2012 Jun 13.
Artigo em Inglês | MEDLINE | ID: mdl-22695027

RESUMO

BACKGROUND: Type 2 diabetes mellitus is a major global public health problem in the worldwide and is increasing in aging populations. Magnesium intake may be one of the most important factors for diabetes prevention and management. Low magnesium intake may exacerbate metabolic abnormalities. In this study, the relationships of magnesium intake with metabolic parameters, depression and physical activity in elderly patients with type 2 diabetes were investigated. METHODS: This cross-sectional study involved 210 type 2 diabetes patients aged 65 years and above. Participants were interviewed to obtain information on lifestyle and 24-hour dietary recall. Assessment of depression was based on DSM-IV criteria. Clinical variables measured included anthropometric measurements, blood pressure, and biochemical determinations of blood and urine samples. Linear regression was applied to determine the relationships of magnesium intake with nutritional variables and metabolic parameters. RESULTS: Among all patients, 88.6% had magnesium intake which was less than the dietary reference intake, and 37.1% had hypomagnesaemia. Metabolic syndromes and depression were associated with lower magnesium intake (p < 0.05). A positive relationship was found between magnesium intake and HDL-cholesterol (p = 0.005). Magnesium intake was inversely correlated with triglyceride, waist circumference, body fat percent and body mass index (p < 0.005). After controlling confounding factor, HDL-cholesterol was significantly higher with increasing quartile of magnesium intake (p for trend = 0005). Waist circumference, body fat percentage, and body mass index were significantly lower with increase quartile of magnesium intake (p for trend < 0.001). The odds of depression, central obesity, high body fat percentage, and high body mass index were significantly lower with increasing quartile of magnesium intake (p for trend < 0.05). In addition, magnesium intake was related to high physical activity level and demonstrated lower serum magnesium levels. Serum magnesium was not significantly associated with metabolic parameters. CONCLUSIONS: The majority of elderly type 2 diabetes who have low magnesium intake may compound this deficiency with metabolic abnormalities and depression. Future studies should determine the effects of increased magnesium intake or magnesium supplementation on metabolic control and depression in elderly people with type 2 diabetes.


Assuntos
Depressão/fisiopatologia , Diabetes Mellitus Tipo 2/fisiopatologia , Magnésio/administração & dosagem , Síndrome Metabólica/fisiopatologia , Atividade Motora , Idoso , Pressão Sanguínea/efeitos dos fármacos , Composição Corporal , Índice de Massa Corporal , HDL-Colesterol/sangue , Estudos Transversais , Depressão/complicações , Diabetes Mellitus Tipo 2/complicações , Feminino , Humanos , Estilo de Vida , Modelos Lineares , Magnésio/sangue , Masculino , Síndrome Metabólica/complicações , Avaliação Nutricional , Razão de Chances , Fatores de Risco , Inquéritos e Questionários , Triglicerídeos/sangue , Circunferência da Cintura
11.
Cytotherapy ; 12(4): 455-65, 2010 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-20230225

RESUMO

BACKGROUND AIMS: Human mesenchymal stromal cells (hMSC) play a crucial role in tissue engineering and regenerative medicine, and have important clinical potential for cell therapy. However, many hMSC studies have been restricted by limited cell numbers and difficult detection in vivo. To expand the lifespan, hMSC are usually immortalized by virus-mediated gene transfer. However, these genetically modified cells easily lose critical phenotypes and stable genotypes because of insertional mutagenesis. METHODS: We used a non-viral transfection method to establish human telomerase reverse transcriptase-immortalized cord blood hMSC (hTERT-cbMSC). We also established red fluorescent protein (RFP)-expressing hTERT-cbMSC (hTERT/RFP-cbMSC) by the same non-viral transfection method, and these cells were injected into a rat model with traumatic brain injury for in vivo detection analysis. RESULTS: The hTERT-cbMSC could grow more than 200 population doublings with a stable doubling time and maintained differentiation capacities. hTERT/RFP-cbMSC could proliferate efficiently within 2 weeks at the injury location and could be detected easily under a fluorescent microscope. Importantly, both hTERT-cbMSC and hTERT/RFP-cbMSC showed no chromosomal abnormalities by karyotype analysis and no tumor formation in severe combined immunodeficient (SCID) mice by transplantation assay. CONCLUSIONS: We have developed immortalized cbMSC with hTERT expression and RFP expression, which will be useful tools for stem cell research and translational study.


Assuntos
Lesões Encefálicas/terapia , Linhagem Celular Transformada , Proteínas Luminescentes/metabolismo , Células-Tronco Mesenquimais , Transplante de Células-Tronco , Células Estromais/metabolismo , Telomerase/metabolismo , Adipogenia/genética , Animais , Lesões Encefálicas/genética , Lesões Encefálicas/patologia , Lesões Encefálicas/fisiopatologia , Imunofluorescência , Perfilação da Expressão Gênica , Humanos , Proteínas Luminescentes/genética , Masculino , Células-Tronco Mesenquimais/metabolismo , Células-Tronco Mesenquimais/patologia , Camundongos , Camundongos SCID , Análise em Microsséries , Modelos Animais , Osteogênese/genética , Ratos , Células Estromais/patologia , Telomerase/genética , Proteína Vermelha Fluorescente
12.
J Biomed Sci ; 17: 9, 2010 Feb 13.
Artigo em Inglês | MEDLINE | ID: mdl-20152049

RESUMO

Although gait change is considered a useful indicator of severity in animal models of Parkinson's disease, systematic and extensive gait analysis in animal models of neurological deficits is not well established. The CatWalk-assisted automated gait analysis system provides a comprehensive way to assess a number of dynamic and static gait parameters simultaneously. In this study, we used the Catwalk system to investigate changes in gait parameters in adult rats with unilateral 6-OHDA-induced lesions and the rescue effect of dopaminergic neuron transplantation on gait function. Four weeks after 6-OHDA injection, the intensity and maximal area of contact were significantly decreased in the affected paws and the swing speed significantly decreased in all four paws. The relative distance between the hind paws also increased, suggesting that animals with unilateral 6-OHDA-induced lesions required all four paws to compensate for loss of balance function. At 8 weeks post-transplantation, engrafted dopaminergic neurons expressed tyrosine hydroxylase. In addition, the intensity, contact area, and swing speed of the four limbs increased and the distance between the hind paws decreased. Partial recovery of methamphetamine-induced rotational response was also noted.


Assuntos
Dopamina/metabolismo , Neurônios/transplante , Doença de Parkinson/terapia , Adrenérgicos/administração & dosagem , Adrenérgicos/farmacologia , Animais , Modelos Animais de Doenças , Células-Tronco Embrionárias/citologia , Feminino , Marcha/fisiologia , Transtornos Neurológicos da Marcha , Neurônios/efeitos dos fármacos , Neurônios/metabolismo , Oxidopamina/administração & dosagem , Oxidopamina/farmacologia , Doença de Parkinson/metabolismo , Doença de Parkinson/patologia , Ratos , Ratos Sprague-Dawley
13.
Cells Tissues Organs ; 192(2): 93-105, 2010.
Artigo em Inglês | MEDLINE | ID: mdl-20215735

RESUMO

The amniotic membrane has been clinically applied as a therapeutic material in wound covering and corneal surface reconstruction. Recently, mesenchymal stem cells (MSCs) have been isolated from the placenta, specifically from the amniotic membrane. However, the localization of MSCs in the amniotic membrane has not been determined. In this study, term placenta was collected, and we performed immunohistochemical staining techniques to identify and localize MSCs in the mesoderm of the amniotic membrane in situ with MSC antibodies, including CD90 and CD105. We further directly cultured and characterized MSCs from the amniotic membrane mesoderm (AMSCs). The AMSCs were easily isolated and represented a homogenous fibroblastic morphology at early passages. In addition to MSC surface markers, AMSCs expressed Sox2, Oct-4 and Nanog. AMSCs could be induced into osteocytes, adipocytes and chondrocytes in vitro and show immunosuppressive effects on T-cell proliferation. Under appropriate conditions, AMSCs could differentiate into neuronal-like cells, which were identified by neuronal-specific markers and their ability to secrete dopamine. This study reveals that AMSCs provide a promising source for stem cell studies and also extend the clinical potential of the amniotic membrane in the field of regenerative medicine.


Assuntos
Âmnio/citologia , Separação Celular/métodos , Células-Tronco Mesenquimais/citologia , Mesoderma/citologia , Neurônios/fisiologia , Adulto , Antígenos CD/metabolismo , Diferenciação Celular , Linhagem da Célula , Condrócitos/citologia , Dopamina/metabolismo , Endoglina , Feminino , Humanos , Células-Tronco Mesenquimais/metabolismo , Osteócitos/citologia , Gravidez , Receptores de Superfície Celular/metabolismo , Antígenos Thy-1/metabolismo
14.
J Surg Res ; 161(1): 101-10, 2010 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-19482304

RESUMO

BACKGROUND: Anastomosis of the nerve especially at narrow surgical field and presence of surgical tension is not easily accessible. DuraSeal demonstrates strong adhesive power without producing neurotoxicity. Herein, we evaluate the possibility of DuraSeal as a substitute in the repair of sciatic nerve gap injury. MATERIALS AND METHODS: The nerve gap model was constructed by excising the sciatic nerve (5mm in length) in Sprague Dawley rats leaving a 5mm nerve defect between nerve stumps. Animals were categorized into four groups: Group I: no treatment; Group II: 4 stitches suture; Group III: nerve approximation fixed by tissue glue; Group IV: nerve approximation fixed by DuraSeal. The motor function assessment included the CatWalk and SFI as well as electrophysiological studies. Nerve continuity and regeneration was examined at 1 and 8 wk after injury. The inflammatory cells, Schwann cell apoptosis, and Schwann cell proliferation were also investigated 1 wk after injury. RESULTS: The achievement of nerve continuity and myelination by DuraSeal approached that of suture demonstrated by crystal violet and Luxol Fast Blue staining at 1 and 8 wk, respectively. Motor function and electrophysiological parameters were restored in DuraSeal and suture group. Early expression of neurofilament and bromodeoxyuridine (BrdU) was also observed in these two groups. There was no statistically significant difference in deposits of macrophages and neutrophil cells or cell apoptosis among these four groups. CONCLUSIONS: DuraSeal achieved the same nerve regeneration compared with that of suture and produced better regeneration than that of the tissue glue or without any treatment. The accomplishment of nerve regeneration and continuity without causing neurotoxicity justifies using DuraSeal as a ligature in the anastomosis of nerve gap injury.


Assuntos
Anastomose Cirúrgica/métodos , Resinas Sintéticas/uso terapêutico , Nervo Isquiático/lesões , Neuropatia Ciática/cirurgia , Adesivos Teciduais/uso terapêutico , Animais , Apoptose , Estimulação Elétrica , Adesivo Tecidual de Fibrina/uso terapêutico , Regeneração Nervosa , Ratos , Ratos Sprague-Dawley , Resinas Sintéticas/toxicidade , Nervo Isquiático/fisiologia , Neuropatia Ciática/imunologia , Técnicas de Sutura
15.
J Sep Sci ; 33(13): 2010-6, 2010 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-20506427

RESUMO

A microdialysis sampling (MDS) on-line SPE (MDS/SPE) has been applied to redeem the detection after dilution to decrease matrix interference in the analysis of ketamine (K) and its two main metabolites, norketamine (NK) and dehydronorketamine (DHNK) in urine by HPLC. After being filtrated, diluted and adjusting the pH, K and its metabolites in the diluted sample solution were collected through MDS and then trapped on an on-line SPE for HPLC analysis. The optimal conditions for MDS/SPE were investigated and then applied to real sample analysis. Experimental results indicated that the MDS/SPE by using regenerated cellulose hollow fiber (8-cm length) and 1 mM sulfuric acid as the perfusate at 20 microL/min flow-rate to collect analytes from 100-fold diluted urine sample (20 mL at pH 6.0), and then having been trapped in octadecyl-modified silica phase SPE for 30 min, offered the optimum efficiency. The concentration levels of 41, 42 and 28% (m/m) for K, NK and DHNK, respectively, in urine were redeemed for determination. The detection limits were 0.38, 0.33 and 0.34 ng/mL (in 100-fold diluted sample) for K, NK and DHNK, respectively. The method provides a very simple, inexpensive and eco-friendly procedure to determine K, NK and DHNK in urine.


Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Ketamina/metabolismo , Ketamina/urina , Microdiálise , Extração em Fase Sólida , Humanos , Masculino , Microdiálise/instrumentação , Valores de Referência , Extração em Fase Sólida/instrumentação , Adulto Jovem
16.
Eur J Appl Physiol ; 108(2): 363-9, 2010 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-19816708

RESUMO

The effect of magnesium supplementation on exercise performance remains controversial. In the present study, the effects of magnesium sulfate on exercise performance and blood glucose metabolism were examined. In order to provide a non-invasive measure of continuous exercise, we developed an auto-blood sampling system was coupled to a microdialysis analyzer to detect the dynamic changes in glucose metabolism in conscious and freely moving gerbils subjected to forced swimming. Gerbils were pretreated with saline or magnesium sulfate (90 mg kg(-1), ip) 30 min before exercise. The duration times were significantly increased by 71% in the magnesium sulfate-treated groups (p < 0.01) when compared with those in the control. Another group of gerbils were subjected to blood sampling assay. A catheter was implanted in the jugular vein of each gerbil for collecting blood samples by the computer-aided blood sampler. The basal levels of plasma glucose, lactate, and magnesium were 6,245 +/- 662, 1,067 +/- 309, and 590 +/- 50 microM, respectively, with no significant difference between groups. Plasma glucose, lactate, and magnesium levels increased to 134 and 204%, 369 and 220%, and 155 and 422% of basal levels during swimming in both the control and magnesium sulfate-treated groups, respectively (p < 0.05). Pretreatment with magnesium sulfate elevated glucose and magnesium levels to 175 and 302% of the basal levels (p < 0.05), respectively, whereas pretreatment with magnesium sulfate reduced the lactate levels 150% of the basal level (p < 0.05) during swimming. Furthermore, the magnesium levels increased to about 152-422% of basal levels during forced swimming and the recovery period (p < 0.05). The present study demonstrates that magnesium sulfate improved the duration time of forced swimming exercise. In addition, magnesium raised glucose levels and attenuated lactate levels during forced swimming. These results indicate that positive effects of magnesium supplementation may contribute to the enhancement of exercise performance in athletes.


Assuntos
Glicemia/metabolismo , Sulfato de Magnésio/farmacologia , Esforço Físico/fisiologia , Animais , Metabolismo Energético , Gerbillinae/metabolismo , Masculino , Microdiálise , Condicionamento Físico Animal/fisiologia , Fatores de Tempo
17.
Chin J Physiol ; 53(5): 299-309, 2010 Oct 31.
Artigo em Inglês | MEDLINE | ID: mdl-21793341

RESUMO

Magnesium sulfate (MgSO4) ameliorates focal ischemia-induced neuronal death in the rat and gerbil models. However, the molecular mechanisms for this neuroprotection are not known. Focal cerebral ischemia was produced by unilateral occlusion of the right common carotid artery and the right middle cerebral artery (CCAO + MCAO) for 30 min or 60 min. Treatment with MgSO4 significantly increased the level of mitogen-activated protein kinase/extra-cellular signal-regulated kinase kinase 1/2 (MEK1/2), extra-cellular signal-regulated kinase 1/2 (ERK1/2), cyclic-AMP response element binding protein (CREB) phosphorylation and the anti-apoptotic protein Bcl-2 both in the non-ischemic (contralateral) and ischemic (ipsilateral) cortex. However, these effects were reversed by administration of U0126, a MEK kinase inhibitor. In the ipsilateral cortex, a significant increase in the level of the proapoptotic proteins Bax, Bad, BNIP3 and activated caspase 3 were detected at the end of focal ischemia compared to the non-ischemic cortex. Treatment of MgSO4 prevented these ischemia-induced activations of the death cascade. Collectively, these data indicate that the ERK-CREB-Bcl-2 signaling pathway might be involved in MgSO4-induced neuroprotection following focal ischemia. Moreover, MgSO4 treatment also resulted in a reduction in pro-apoptotic proteins. These results enhance our understanding on the role of MgSO4 in treating cerebral ischemia.


Assuntos
Isquemia Encefálica/metabolismo , Isquemia Encefálica/prevenção & controle , Córtex Cerebral/metabolismo , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Sulfato de Magnésio/uso terapêutico , Transdução de Sinais/fisiologia , Animais , Proteínas Reguladoras de Apoptose/metabolismo , Isquemia Encefálica/patologia , Caspase 3/metabolismo , Córtex Cerebral/efeitos dos fármacos , Córtex Cerebral/patologia , Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/metabolismo , Gerbillinae , Sulfato de Magnésio/farmacologia , Masculino , Modelos Animais , Neurônios/efeitos dos fármacos , Neurônios/metabolismo , Neurônios/patologia , Fosforilação/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Transdução de Sinais/efeitos dos fármacos
18.
Biochem Biophys Res Commun ; 382(1): 177-82, 2009 Apr 24.
Artigo em Inglês | MEDLINE | ID: mdl-19275877

RESUMO

Granulocyte colony-stimulating factor (G-CSF) demonstrates neuroprotective effects through different mechanisms, including mobilization of bone marrow cells. However, the influence of G-CSF-mediated mobilization of bone marrow-derived cells on injured sciatic nerves remains to be elucidated. The administration of G-CSF promoted a short-term functional recovery 7 days after crush injury in sciatic nerves. A double-immunofluorescence study using green fluorescent protein-chimeric mice revealed that bone marrow-derived CD34+ cells were predominantly mobilized and migrated into injured nerves after G-CSF treatment. G-CSF-mediated beneficial effects against sciatic nerve injury were associated with increased CD34+ cell deposition, vascular endothelial growth factor (VEGF) expression, and vascularization/angiogenesis as well as decreased CD68+ cell accumulation. However, cell differentiation and VEGF expression were not demonstrated in deposited cells. The results suggest that the promotion of short-term functional recovery in sciatic nerve crush injury by G-CSF involves a paracrine modulatory effect and a bone marrow-derived CD34+ cell mobilizing effect.


Assuntos
Fator Estimulador de Colônias de Granulócitos/farmacologia , Neovascularização Fisiológica/efeitos dos fármacos , Regeneração Nervosa/efeitos dos fármacos , Fármacos Neuroprotetores/farmacologia , Nervo Isquiático/efeitos dos fármacos , Nervo Isquiático/lesões , Animais , Antígenos CD34/análise , Células da Medula Óssea/efeitos dos fármacos , Células da Medula Óssea/fisiologia , Movimento Celular/efeitos dos fármacos , Proteínas de Fluorescência Verde/genética , Masculino , Camundongos , Camundongos Transgênicos , Nervo Isquiático/fisiologia
19.
J Biomed Sci ; 16: 75, 2009 Aug 23.
Artigo em Inglês | MEDLINE | ID: mdl-19698158

RESUMO

Attenuation of inflammatory cell deposits and associated cytokines prevented the apoptosis of transplanted stem cells in a sciatic nerve crush injury model. Suppression of inflammatory cytokines by fermented soybean extracts (Natto) was also beneficial to nerve regeneration. In this study, the effect of Natto on transplanted human amniotic fluid mesenchymal stem cells (AFS) was evaluated. Peripheral nerve injury was induced in SD rats by crushing a sciatic nerve using a vessel clamp. Animals were categorized into four groups: Group I: no treatment; Group II: fed with Natto (16 mg/day for 7 consecutive days); Group III: AFS embedded in fibrin glue; Group IV: Combination of group II and III therapy. Transplanted AFS and Schwann cell apoptosis, inflammatory cell deposits and associated cytokines, motor function, and nerve regeneration were evaluated 7 or 28 days after injury. The deterioration of neurological function was attenuated by AFS, Natto, or the combined therapy. The combined therapy caused the most significantly beneficial effects. Administration of Natto suppressed the inflammatory responses and correlated with decreased AFS and Schwann cell apoptosis. The decreased AFS apoptosis was in line with neurological improvement such as expression of early regeneration marker of neurofilament and late markers of S-100 and decreased vacuole formation. Administration of either AFS, or Natto, or combined therapy augmented the nerve regeneration. In conclusion, administration of Natto may rescue the AFS and Schwann cells from apoptosis by suppressing the macrophage deposits, associated inflammatory cytokines, and fibrin deposits.


Assuntos
Transplante de Células-Tronco Mesenquimais , Compressão Nervosa/reabilitação , Regeneração Nervosa/efeitos dos fármacos , Fitoterapia , Extratos Vegetais/uso terapêutico , Nervo Isquiático/efeitos dos fármacos , Alimentos de Soja , Líquido Amniótico/citologia , Animais , Apoptose/efeitos dos fármacos , Citocinas/antagonistas & inibidores , Citocinas/fisiologia , Fibrina/análise , Adesivo Tecidual de Fibrina/toxicidade , Inflamação/patologia , Macrófagos/efeitos dos fármacos , Proteínas do Tecido Nervoso/análise , Extratos Vegetais/administração & dosagem , Ratos , Ratos Sprague-Dawley , Recuperação de Função Fisiológica , Células de Schwann/efeitos dos fármacos , Células de Schwann/patologia , Nervo Isquiático/fisiologia
20.
Neurochem Res ; 34(7): 1304-16, 2009 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-19152028

RESUMO

PURPOSE: Attenuation of pro-inflammatory cytokines and associated inflammatory cell deposits rescues human amniotic fluid mesenchymal stem cells (AFS) from apoptosis. Hyperbaric oxygen (HBO) suppressed stimulus-induced pro-inflammatory cytokine production in blood-derived monocyte-macrophages. Herein, we evaluate the beneficial effect of hyperbaric oxygen on transplanted AFS in a sciatic nerve injury model. METHODS: Peripheral nerve injury was produced in Sprague-Dawley rats by crushing the left sciatic nerve using a vessel clamp. The AFS were embedded in fibrin glue and delivered to the injured site. Hyperbaric oxygen (100% oxygen, 2 ATA, 60 min/day) was administered 12 h after operation for seven consecutive days. Transplanted cell apoptosis, oxidative stress, inflammatory cell deposits and associated chemokines, pro-inflammatory cytokines, motor function, and nerve regeneration were evaluated 7 and 28 days after injury. RESULTS: Crush injury induced an inflammatory response, disrupted nerve integrity, and impaired nerve function in the sciatic nerve. However, crush injury-provoked inflammatory cytokines, deposits of inflammatory cytokines, and associated macrophage migration chemokines were attenuated in groups receiving hyperbaric oxygen but not in the AFS-only group. No significant increase in oxidative stress was observed after administration of HBO. In transplanted AFS, marked apoptosis was detected and this event was reduced by HBO treatment. Increased nerve myelination and improved motor function were observed in AFS-transplant, HBO-administrated, and AFS/HBO-combined treatment groups. Significantly, the AFS/HBO combined treatment showed the most beneficial effect. CONCLUSION: AFS in combination with HBO augment peripheral nerve regeneration, which may involve the suppression of apoptotic death in implanted AFS and the attenuation of an inflammatory response detrimental to peripheral nerve regeneration.


Assuntos
Oxigenoterapia Hiperbárica , Transplante de Células-Tronco Mesenquimais , Regeneração Nervosa/fisiologia , Neuropatia Ciática/terapia , Líquido Amniótico/citologia , Animais , Apoptose/efeitos dos fármacos , Quimiocina CCL2/metabolismo , Quimiocina CCL5/metabolismo , Eletrofisiologia , Humanos , Macrófagos/fisiologia , Modelos Animais , Regeneração Nervosa/efeitos dos fármacos , Nervos Periféricos/fisiologia , Peroxidase/metabolismo , Ratos , Ratos Sprague-Dawley , Nervo Isquiático/lesões , Nervo Isquiático/fisiologia
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