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1.
World J Clin Cases ; 9(20): 5547-5555, 2021 Jul 16.
Artigo em Inglês | MEDLINE | ID: mdl-34307608

RESUMO

BACKGROUND: The Hartmann procedure is currently recognized as a common, safe, and feasible surgical procedure. However, its reversal rate is low, and the optimal timing for Hartmann reversal surgery is controversial. CASE SUMMARY: A 65-year-old man came to our hospital with a complaint of an intestinal fistula next to the stoma. The patient had undergone a Hartmann procedure 13 years prior. We performed colonoscopy, computed tomography, and other diagnostics before successfully reversing the stoma. CONCLUSION: Although the optimal time for Hartmann procedure reversal is controversial, time may ultimately not be a factor in the success of reversal.

2.
World J Gastrointest Surg ; 13(3): 303-314, 2021 Mar 27.
Artigo em Inglês | MEDLINE | ID: mdl-33796217

RESUMO

BACKGROUND: With advancements in laparoscopic technology and the wide application of linear staplers, sphincter-saving procedures are increasingly performed for low rectal cancer. However, sphincter-saving procedures have led to the emergence of a unique clinical disorder termed anterior rectal resection syndrome. Colonic pouch anastomosis improves the quality of life of patients with rectal cancer > 7 cm from the anal margin. But whether colonic pouch anastomosis can reduce the incidence of rectal resection syndrome in patients with low rectal cancer is unknown. AIM: To compare postoperative and oncological outcomes and bowel function of straight and colonic pouch anal anastomoses after resection of low rectal cancer. METHODS: We conducted a retrospective study of 72 patients with low rectal cancer who underwent sphincter-saving procedures with either straight or colonic pouch anastomoses. Functional evaluations were completed preoperatively and at 1, 6, and 12 mo postoperatively. We also compared perioperative and oncological outcomes between two groups that had undergone low or ultralow anterior rectal resection. RESULTS: There were no significant differences in mean operating time, blood loss, time to first passage of flatus and excrement, and duration of hospital stay between the colonic pouch and straight anastomosis groups. The incidence of anastomotic leakage following colonic pouch construction was lower (11.4% vs 16.2%) but not significantly different than that of straight anastomosis. Patients with colonic pouch construction had lower postoperative low anterior resection syndrome scores than the straight anastomosis group, suggesting better bowel function (preoperative: 4.71 vs 3.89, P = 0.43; 1 mo after surgery: 34.2 vs 34.7, P = 0.59; 6 mo after surgery: 22.70 vs 29.0, P < 0.05; 12 mo after surgery: 15.5 vs 19.5, P = 0.01). The overall recurrence and metastasis rates were similar (4.3% and 11.4%, respectively). CONCLUSION: Colonic pouch anastomosis is a safe and effective procedure for colorectal reconstruction after low and ultralow rectal resections. Moreover, colonic pouch construction may provide better functional outcomes compared to straight anastomosis.

3.
World J Clin Cases ; 8(23): 6095-6102, 2020 Dec 06.
Artigo em Inglês | MEDLINE | ID: mdl-33344610

RESUMO

BACKGROUND: Small-cell neuroendocrine carcinoma (SNEC) of the rectum is a rare tumor associated with poor prognosis. CASE SUMMARY: We report a case of a 77-year-old male who came into our hospital because of blood with his stool. An endoscopy revealed a cauliflower-like neoplasm in his rectum. Imaging examination showed that the lesion in the upper rectum was likely rectal cancer, and there was no evidence of metastasis. The patient was treated with surgery. Pathological examination confirmed SNEC of the rectum and an R0 resection was achieved. However, 1 mo after the operation, the patient developed intestinal and ureteral obstructions due to peritoneal metastases. Finally, the patient died from renal failure. CONCLUSION: SNEC of the rectum is a high-grade carcinoma with an aggressive phenotype, and surgery should be cautiously considered.

4.
Stem Cell Res ; 38: 101464, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-31176915

RESUMO

The in vitro expansion of endothelial progenitor cells (EPCs) is necessary for obtaining sufficient amounts of cells for clinical applications. However, EPC expansion is conventionally carried out under non-physiologic oxygen concentrations (normoxia, ~20% O2). We compared the effects of normoxic and hypoxic culture on the stemness of expanded EPCs. Human EPCs were cultured under hypoxia (1% O2) or normoxia (~20% O2), respectively. Cell proliferation, colony formation, in vitro angiogenesis, and the migration ability of the expanded EPCs were compared. To explore the underlying mechanism, whole transcriptome RNA sequencing (RNA-seq) was also performed to select differentially expressed genes (DEGs), which were then partially validated by western blotting. EPCs cultured under normoxia showed reduced proliferation, colony formation, in vitro angiogenesis, and migration abilities and a higher proportion of senescent cells compared with those cultured under hypoxia. A total of 48 DEGs were identified by transcriptome RNA-seq. Further bioinformatics analysis revealed that six pathways were enriched, among which the p53 signaling pathway. Finally, we confirmed the differential expression of the p53 pathway by Western blot analysis. Compared with hypoxia, normoxia is not favorable for maintaining the stemness of human EPCs. Several signaling pathways, including the p53 signaling pathway implicate in reducing stemness of EPCs under normoxia.


Assuntos
Proliferação de Células , Senescência Celular , Células Progenitoras Endoteliais/metabolismo , Neovascularização Fisiológica , Transdução de Sinais , Adulto , Idoso , Hipóxia Celular , Células Cultivadas , Células Progenitoras Endoteliais/citologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade
5.
Stem Cells Transl Med ; 8(1): 14-21, 2019 01.
Artigo em Inglês | MEDLINE | ID: mdl-30156755

RESUMO

Transplantation of endothelial progenitor cells (EPCs) is a proven safe and effective method for treatment of cerebral ischemia in animal experiments. However, safety and efficacy need to be determined in clinical trials. We performed a two-center, randomized, placebo-controlled phase I/IIa trial with blinded outcome assessment on 18 patients with acute cerebral infarct within the middle cerebral artery territory, and followed for up to 4 years. Autologous ex vivo expanded EPCs were injected intravenously in the EPC group, and patients who received saline or autologous bone marrow stromal cells served as control groups. Mortality of any cause, adverse events, and new-onset comorbidities were monitored. Changes in neurological deficits were assessed at different time points. We found no toxicity events or infusional or allergic reactions in any treated group. Three patients in the placebo group died during the 4-year follow-up. We found that the EPC group had fewer serious adverse events compared with the placebo-controlled group, although there were no statistical differences in mortality among the three groups. Furthermore, there was no significant difference in neurological or functional improvement observed among the three groups, except for the Scandinavia Stroke Scale score at 3 months between the EPC group and placebo-controlled group. Autologous transplantation of EPCs appears to improve long-term safety in acute cerebral infarct patients, supporting the feasibility of this novel method for treatment of ischemic stroke (ClinicalTrials.gov: NCT01468064). Stem Cells Translational Medicine 2019;8:14-21.


Assuntos
Células Progenitoras Endoteliais/citologia , Células-Tronco Mesenquimais/citologia , Acidente Vascular Cerebral/terapia , Adulto , Idoso , Sobrevivência Celular/fisiologia , Células Progenitoras Endoteliais/transplante , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Transplante Autólogo
6.
eNeuro ; 5(3)2018.
Artigo em Inglês | MEDLINE | ID: mdl-29963617

RESUMO

Acute ischemic stroke (AIS) is caused by clotting in the cerebral arteries, leading to brain oxygen deprivation and cerebral infarction. Recombinant human tissue plasminogen activator (tPA) is currently the only Food and Drug Administration-approved drug for ischemic stroke. However, tPA has to be administered within 4.5 h from the disease onset and delayed treatment of tPA can increase the risk of neurovascular impairment, including neuronal cell death, blood-brain barrier (BBB) disruption, and hemorrhagic transformation. A key contributing factor for tPA-induced neurovascular impairment is activation of matrix metalloproteinase-9 (MMP-9). We used a clinically-relevant mouse embolic model of focal-cerebral ischemia by insertion of a single embolus of blood clot to block the right middle cerebral artery. We showed that administration of the potent and highly selective gelatinase inhibitor SB-3CT extends the time window for administration of tPA, attenuating infarct volume, mitigating BBB disruption, and antagonizing the increase in cerebral hemorrhage induced by tPA treatment. We demonstrated that SB-3CT attenuates tPA-induced expression of vascular MMP-9, prevents gelatinase-mediated cleavage of extracellular laminin, rescues endothelial cells, and reduces caveolae-mediated transcytosis of endothelial cells. These results suggest that abrogation of MMP-9 activity mitigates the detrimental effects of tPA treatment, thus the combination treatment holds great promise for extending the therapeutic window for tPA thrombolysis, which opens the opportunity for clinical recourse to a greater number of patients.


Assuntos
Isquemia Encefálica/enzimologia , Gelatinases/metabolismo , Ativador de Plasminogênio Tecidual/administração & dosagem , Animais , Barreira Hematoencefálica/metabolismo , Isquemia Encefálica/prevenção & controle , Caveolina 1/metabolismo , Modelos Animais de Doenças , Células Endoteliais/efeitos dos fármacos , Células Endoteliais/metabolismo , Gelatinases/antagonistas & inibidores , Compostos Heterocíclicos com 1 Anel/administração & dosagem , Laminina/metabolismo , Masculino , Metaloproteinase 9 da Matriz/metabolismo , Inibidores de Metaloproteinases de Matriz/administração & dosagem , Camundongos Endogâmicos C57BL , Sulfonas/administração & dosagem
7.
Brain Inj ; 32(3): 342-349, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29333886

RESUMO

OBJECTIVE: To determine early effects on outcome from traumatic brain injury (TBI) induced by controlled cortical impact (CCI) associated with anaemia in mice. HYPOTHESIS: Outcome from TBI with concomitant anaemia would be worse than TBI without anaemia. METHODS: CCI was induced with electromagnetic impaction in four groups of C57BL/6J mice: sham, sham+anaemia; TBI; and TBI+anaemia. Anaemia was created by withdrawal of 30% of calculated intravascular blood volume and saline replacement of equal volume. Functional outcome was assessed by beam-walking test and open field test (after pre-injury training) on post-injury days 3 and 7. After functional assessment, brains removed from sacrificed animals were pathological reviewed with haematoxylin and eosin, cresyl violet, Luxol Fast Blue, and IBA-1 immunostains. RESULTS: Beam-walking was similar between animals with TBI and TBI+anaemia (p = 0.9). In open field test, animals with TBI+anaemia walked less distance than TBI alone or sham animals on days 3 (p < 0.001) and 7 (p < 0.05), indicating less exploratory and locomotion behaviours. No specific pathologic differences could be identified. CONCLUSIONS: Anaemia associated with TBI from CCI is associated with worse outcome as measured by less distance travelled in the open field test at three days than if anaemia is not present.


Assuntos
Anemia/etiologia , Lesões Encefálicas Traumáticas/complicações , Anemia/patologia , Animais , Encéfalo/metabolismo , Encéfalo/patologia , Proteínas de Ligação ao Cálcio/metabolismo , Modelos Animais de Doenças , Progressão da Doença , Comportamento Exploratório/fisiologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Proteínas dos Microfilamentos/metabolismo , Avaliação de Resultados em Cuidados de Saúde , Transtornos Psicomotores/diagnóstico , Transtornos Psicomotores/etiologia , Estatísticas não Paramétricas
8.
Methods Mol Biol ; 1626: 147-155, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28608207

RESUMO

Pathological activation of gelatinases (matrix metalloproteinase-2 and -9; MMP-2/-9) has been shown to cause a number of detrimental outcomes in neurodegenerative diseases. In gel gelatin zymography is a highly sensitive methodology commonly used in revealing levels of gelatinase activity and in separating the proform and active form of gelatinases, based on their different molecular weights. However, this methodology is inadequate in resolving complex enzyme isoforms, because gelatinase expression and activity can be regulated at transcriptional and/or post-translational levels under in vivo conditions resulting in alternation of their isoelectric focusing (IEF) points. In this chapter, we describe an advanced methodology, termed two-dimensional zymography, combining IEF with zymographic electrophoresis under non-reducing conditions to achieve significant improvement in separation of the gelatinase isoforms in both cell-based and in vivo models for acute brain injuries and neuroinflammation.


Assuntos
Eletroforese em Gel de Poliacrilamida/métodos , Ensaios Enzimáticos/métodos , Gelatina/metabolismo , Gelatinases/metabolismo , Focalização Isoelétrica/métodos , Doenças Neurodegenerativas/enzimologia , Animais , Encéfalo/enzimologia , Encéfalo/metabolismo , Modelos Animais de Doenças , Ativação Enzimática , Gelatinases/análise , Gelatinases/isolamento & purificação , Camundongos , Doenças Neurodegenerativas/metabolismo , Isoformas de Proteínas/análise , Isoformas de Proteínas/metabolismo , Ratos
9.
Medicine (Baltimore) ; 96(20): e6775, 2017 May.
Artigo em Inglês | MEDLINE | ID: mdl-28514293

RESUMO

Rheumatoid arthritis (RA) is polygenic autoimmune disease with unclear etiology. MicroRNAs (miRNAs) play a critical role in the pathogenesis of RA. The objective of this study was to evaluate the role of miR-146a in patients with RA receiving Tripterygium wilfordii Hook F (TwHF) treatment.In total, 69 patients with RA and 69 healthy controls (HC) were included in the study, and patients with RA received TwHF treatment for 24 weeks. Blood samples were collected from RA patients and HC, and peripheral blood mononuclear cells (PBMCs) were isolated. Expression of miR-146a was analyzed in RA patients (baseline, 12 weeks and 24 weeks) and HC.Circulating miR-146a expression was markedly increased in patients with RA compared with healthy controls (P < .001), ROC analysis of miR-146a for diagnosis for RA showed that the AUC was 0.908 (95% CI: 0.862-0.955) with a sensitivity of 87.0% and a specificity of 82.6% at best cutoff. And miR-146a expression was positively associated with the DAS28 score and CRP level (P = .002 and P = .019). Moreover, miR-146a expression was markedly reduced after TwHF therapy (P < .001), and baseline miR-146a level was observed to present an increased tendency in responders compared with non-responders at 24 weeks (P = .066).Our study presented that circulating miR-146a level was correlated with risk and disease activity of RA patients by TwHF treatment, which could strikingly decrease expression of miR-146a in RA patients, and miR-146a may have a value in predicting clinical response of TwHF treatment. It indicates that circulating miR-146a plays a prominent role in RA patients treated by TwHF.


Assuntos
Antirreumáticos/uso terapêutico , Artrite Reumatoide/sangue , Artrite Reumatoide/tratamento farmacológico , Medicamentos de Ervas Chinesas/uso terapêutico , MicroRNAs/sangue , Fitoterapia , Tripterygium , Área Sob a Curva , Biomarcadores/sangue , Feminino , Humanos , Leucócitos Mononucleares/metabolismo , Masculino , Pessoa de Meia-Idade , Prognóstico , Curva ROC , Risco , Índice de Gravidade de Doença , Resultado do Tratamento
10.
Biotechnol Lett ; 38(5): 893-900, 2016 May.
Artigo em Inglês | MEDLINE | ID: mdl-26910777

RESUMO

OBJECTIVE: To test the feasibility of secretion of functional chondroitinase ABC (ChABC), a bacterial enzyme that promotes axonal regeneration after spinal cord injury, from human bone marrow stromal cells (hBMSCs). RESULTS: A lentiviral-expression vector, Lenti6.3-ChABC-3F, carrying the ChABC-3F gene without the bacterial leader sequence (aa 1-24) was constructed. Transfection of these Lenti6.3-ChABC-3F lentivirus led to stable expression in and secrection of ChABC proteins from hBMSCs for at least ten passages in culture in vitro, which was demonstrated by QRT-PCR, immunostaining, western blotting and ELISA. Moreover, the secreted ChABC proteins exhibited similar functional activity as the commercially-available ChABC. CONCLUSIONS: The lentivirus-mediated transfection of chondroitinase ABC gene without the bacterial leader sequence induced substantial long-term secretion of functional ChABC in hBMSCs.


Assuntos
Condroitina ABC Liase/metabolismo , Células-Tronco Mesenquimais/metabolismo , Proteínas Recombinantes/metabolismo , Células Cultivadas , Condroitina ABC Liase/genética , Expressão Gênica , Perfilação da Expressão Gênica , Instabilidade Genômica , Humanos , Imunoensaio , Lentivirus/genética , Sinais Direcionadores de Proteínas/genética , Reação em Cadeia da Polimerase em Tempo Real , Proteínas Recombinantes/genética , Transfecção
11.
ACS Chem Neurosci ; 6(10): 1658-64, 2015 Oct 21.
Artigo em Inglês | MEDLINE | ID: mdl-26241578

RESUMO

SB-3CT is a potent and selective inhibitor of matrix metalloproteinase (MMP)-2 and -9, which has shown efficacy in an animal model of severe traumatic brain injury (TBI). However, SB-3CT is poorly water-soluble and is metabolized primarily to p-hydroxy SB-3CT (2), a more potent inhibitor than SB-3CT. We synthesized the O-phosphate prodrug (3) of compound 2 to enhance its water solubility by more than 2000-fold. The prodrug 3 was a poor MMP inhibitor, but readily hydrolyzed to the active 2 in human blood. Pharmacokinetics and brain distribution studies in mice showed that 2 crossed the blood-brain barrier (BBB) and achieved therapeutic concentrations in the brain. The prodrug 3/compound 2 was evaluated in a mouse model of severe TBI and found to significantly decrease the brain lesion volume and improve neurological outcomes. MMP-9 inhibition by a water-soluble thiirane inhibitor is a promising therapy for treatment of TBI.


Assuntos
Lesões Encefálicas/tratamento farmacológico , Lesões Encefálicas/patologia , Compostos Heterocíclicos com 1 Anel/uso terapêutico , Inibidores de Metaloproteinases de Matriz/uso terapêutico , Sulfonas/uso terapêutico , Animais , Área Sob a Curva , Barreira Hematoencefálica/efeitos dos fármacos , Barreira Hematoencefálica/metabolismo , Lesões Encefálicas/fisiopatologia , Linhagem Celular , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Compostos Heterocíclicos com 1 Anel/farmacologia , Concentração Inibidora 50 , Masculino , Metaloproteinase 9 da Matriz/metabolismo , Inibidores de Metaloproteinases de Matriz/química , Inibidores de Metaloproteinases de Matriz/farmacologia , Camundongos , Camundongos Endogâmicos C57BL , Exame Neurológico , Desempenho Psicomotor/efeitos dos fármacos , Solubilidade , Sulfonas/farmacologia , Água/metabolismo
12.
PLoS One ; 10(4): e0123852, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25859655

RESUMO

Excessive activation of gelatinases (MMP-2/-9) is a key cause of detrimental outcomes in neurodegenerative diseases. A single-dimension zymography has been widely used to determine gelatinase expression and activity, but this method is inadequate in resolving complex enzyme isoforms, because gelatinase expression and activity could be modified at transcriptional and posttranslational levels. In this study, we investigated gelatinase isoforms under in vitro and in vivo conditions using two-dimensional (2D) gelatin zymography electrophoresis, a protocol allowing separation of proteins based on isoelectric points (pI) and molecular weights. We observed organomercuric chemical 4-aminophenylmercuric acetate-induced activation of MMP-2 isoforms with variant pI values in the conditioned medium of human fibrosarcoma HT1080 cells. Studies with murine BV-2 microglial cells indicated a series of proform MMP-9 spots separated by variant pI values due to stimulation with lipopolysaccharide (LPS). The MMP-9 pI values were shifted after treatment with alkaline phosphatase, suggesting presence of phosphorylated isoforms due to the proinflammatory stimulation. Similar MMP-9 isoforms with variant pI values in the same molecular weight were also found in mouse brains after ischemic and traumatic brain injuries. In contrast, there was no detectable pI differentiation of MMP-9 in the brains of chronic Zucker obese rats. These results demonstrated effective use of 2D zymography to separate modified MMP isoforms with variant pI values and to detect posttranslational modifications under different pathological conditions.


Assuntos
Lesões Encefálicas/enzimologia , Gelatinases/metabolismo , Microglia/metabolismo , Animais , Encéfalo/metabolismo , Encéfalo/patologia , Lesões Encefálicas/diagnóstico , Isquemia Encefálica/metabolismo , Isquemia Encefálica/patologia , Linhagem Celular , Meios de Cultivo Condicionados/metabolismo , Modelos Animais de Doenças , Humanos , Isoenzimas , Masculino , Metaloproteinase 9 da Matriz/metabolismo , Camundongos , Inflamação Neurogênica/metabolismo , Ratos
13.
PLoS One ; 9(11): e113531, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25420111

RESUMO

Aged garlic extract (AGE) is widely used as a dietary supplement, and is claimed to promote human health through anti-oxidant/anti-inflammatory activities with hypolipidemic, antiplatelet and neuroprotective effects. Prior studies of AGE have mainly focused on its organosulfur compounds, with little attention paid to its carbohydrate derivatives, such as N-α-(1-deoxy-D-fructos-1-yl)-L-arginine (FruArg). The goal of this study is to investigate actions of AGE and FruArg on antioxidative and neuroinflammatory responses in lipopolysaccharide (LPS)-activated murine BV-2 microglial cells using a proteomic approach. Our data show that both AGE and FruArg can significantly inhibit LPS-induced nitric oxide (NO) production in BV-2 cells. Quantitative proteomic analysis by combining two dimensional differential in-gel electrophoresis (2D-DIGE) with mass spectrometry revealed that expressions of 26 proteins were significantly altered upon LPS exposure, while levels of 20 and 21 proteins exhibited significant changes in response to AGE and FruArg treatments, respectively, in LPS-stimulated BV-2 cells. Notably, approximate 78% of the proteins responding to AGE and FruArg treatments are in common, suggesting that FruArg is a major active component of AGE. MULTICOM-PDCN and Ingenuity Pathway Analyses indicate that the proteins differentially affected by treatment with AGE and FruArg are involved in inflammatory responses and the Nrf2-mediated oxidative stress response. Collectively, these results suggest that AGE and FruArg attenuate neuroinflammatory responses and promote resilience in LPS-activated BV-2 cells by suppressing NO production and by regulating expression of multiple protein targets associated with oxidative stress.


Assuntos
Alho/química , Lipopolissacarídeos/farmacologia , Microglia/efeitos dos fármacos , Extratos Vegetais/farmacologia , Proteoma/análise , Proteômica , Animais , Anti-Inflamatórios/farmacologia , Western Blotting , Linhagem Celular , Dipeptídeos/química , Dipeptídeos/farmacologia , Relação Dose-Resposta a Droga , Eletroforese em Gel Bidimensional , Camundongos , Microglia/citologia , Microglia/metabolismo , Estrutura Molecular , Óxido Nítrico/antagonistas & inibidores , Óxido Nítrico/metabolismo , Transdução de Sinais/efeitos dos fármacos , Espectrometria de Massas em Tandem , Fatores de Tempo
14.
PLoS One ; 9(3): e92133, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24637518

RESUMO

Quantitative assessment of serial brain sections provides an objective measure of neurological events at cellular and molecular levels but is difficult to implement in experimental neuroscience laboratories because of variation from person-to-person and the time required for analysis. Whole slide imaging (WSI) technology, recently introduced for pathological diagnoses, offers an electronic environment and a variety of computational tools for performing high-throughput histological analysis and managing the associated information. In our study, we applied various algorithms to quantify histologic changes associated with brain injury and compared the results to manual assessment. WSI showed a high degree of concordance with manual quantitation by Pearson correlation and strong agreement using Bland-Altman plots in: (i) cortical necrosis in cresyl-violet-stained brain sections of mice after focal cerebral ischemia; (ii) intracerebral hemorrhage in ischemic mouse brains for automated annotation of the small regions, rather than whole hemisphere of the tissue sections; (iii) Iba1-immunoreactive cell density in the adjacent and remote brain regions of mice subject to controlled cortical impact (CCI); and (iv) neuronal degeneration by silver staining after CCI. These results show that WSI, when appropriately applied and carefully validated, is a highly efficient and unbiased tool to locate and identify neuropathological features, delineate affected regions and histologically quantify these events.


Assuntos
Lesões Encefálicas/patologia , Neuroimagem/métodos , Algoritmos , Animais , Automação , Lesões Encefálicas/complicações , Contagem de Células , Córtex Cerebral/patologia , Hemorragia Cerebral/complicações , Hemorragia Cerebral/patologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Microglia/patologia , Necrose , Degeneração Neural/complicações , Degeneração Neural/patologia , Projetos Piloto , Coloração pela Prata
15.
Biochem Biophys Res Commun ; 440(4): 502-8, 2013 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-24055873

RESUMO

Previous studies have confirmed the therapeutic effects of bone marrow stromal cells (BMSCs) transplantation on cerebral ischemia. However, the proliferative, differentiative, and homing capacity of BMSC from the elderly are significantly reduced, especially after several passages expansion in vitro. In this study, by introducing lentivirus-mediated hTERT and VEGF genes to modify human BMSCs from aged donors, we observed extended lifespan, promoted angiogenic capacity while less enhanced tumorigenicity of the genetically engineering BMSCs. These results therefore suggest that the modification of aged BMSCs by dual expression of hTERT and VEGF may be used for autologous cell replacement for ischemic cerebrovascular disease in elderly patients.


Assuntos
Células da Medula Óssea/fisiologia , Senescência Celular , Neovascularização Fisiológica , Telomerase/biossíntese , Fator A de Crescimento do Endotélio Vascular/biossíntese , Idoso , Células da Medula Óssea/citologia , Transplante de Medula Óssea , Engenharia Celular , Engenharia Genética , Humanos , Acidente Vascular Cerebral/terapia , Células Estromais/citologia , Células Estromais/fisiologia , Telomerase/genética , Fator A de Crescimento do Endotélio Vascular/genética
16.
BMC Neurosci ; 14: 24, 2013 Mar 04.
Artigo em Inglês | MEDLINE | ID: mdl-23452440

RESUMO

BACKGROUND: Post-stroke hyperglycemia appears to be associated with poor outcome from stroke, greater mortality, and reduced functional recovery. Focal cerebral ischemia data support that neural stem cells (NSCs) play an important role in post-ischemic repair. Here we sought to evaluate the negative effects of hyperglycemia on the cellular biology of NSCs following anoxia, and to test whether high glucose affects NSC recovery from ischemic injury. RESULTS: In this study, we used immortalized adult neural stem cells lines and we induced in vitro ischemia by 6 h oxygen and glucose deprivation (OGD) in an anaerobic incubator. Reperfusion was performed by returning cells to normoxic conditions and the cells were then incubated in experimental medium with various concentrations of glucose (17.5, 27.75, 41.75, and 83.75 mM) for 24 h. We found that high glucose (≥27.75 mM) exposure induced apoptosis of NSCs in a dose-dependent manner after exposure to OGD, using an Annexin V/PI apoptosis detection kit. The cell viability and proliferative activity of NSCs following OGD in vitro, evaluated with both a Cell Counting kit-8 (CCK-8) assay and a 5-ethynyl-2'-deoxyuridine (EdU) incorporation assay, were inhibited by high glucose exposure. Cell cycle analysis showed that high glucose exposure increased the percentage of cells in G0/G1-phase, and reduced the percentage of cells in S-phase. Furthermore, high glucose exposure was found to significantly induce the activation of c-Jun N-terminal protein kinase (JNK) and p38 mitogen-activated protein kinase (MAPK) and suppress extracellular signal-regulated kinase 1/2 (ERK1/2) activity. CONCLUSIONS: Our results demonstrate that high glucose induces apoptosis and inhibits proliferation of NSCs following OGD in vitro, which may be associated with the activation of JNK/p38 MAPK pathways and the delay of G1-S transition in the cells.


Assuntos
Células-Tronco Adultas/efeitos dos fármacos , Apoptose/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Glucose/farmacologia , Animais , Ciclo Celular/efeitos dos fármacos , Sobrevivência Celular , Células Cultivadas , Desoxiuridina/análogos & derivados , Desoxiuridina/metabolismo , Relação Dose-Resposta a Droga , Glucose/deficiência , Hipóxia , Necrose/tratamento farmacológico , Proteínas do Tecido Nervoso/metabolismo , Ratos
17.
Nan Fang Yi Ke Da Xue Xue Bao ; 32(5): 659-63, 2012 May.
Artigo em Chinês | MEDLINE | ID: mdl-22588919

RESUMO

OBJECTIVE: To investigate the dynamic changes in CD4(+)CD25(+) regulatory T cells and Foxp3 expression in peripheral blood and brain tissues of rats after acute cerebral ischemia and explore their role in the pathophysiological evolution of acute ischemic stroke. METHODS: Forty-eight Wistar rats were randomized equally into ischemia and sham-operated groups, and right middle cerebral artery occlusion was induced in the former group. Flow cytometry and immunohistochemistry were employed to detect CD4(+)CD25(+) T cells and Foxp3 expression, respectively, in the peripheral blood and brain tissue at 1, 3, 7, and 14 days after modeling. The behavioral changes of the rats were evaluated using an improved NSS neurological functional scoring system. RESULTS: The neurological function scores of the two groups both gradually declined after the operation, and showed significant differences between the two groups at all the time points of measurement (P<0.01). The CD4(+)CD25 T cells in the peripheral blood were similar between the two group at 1 and 3 days after the operation (P>0.05), but increased significantly in the ischemia group at 7 and 14 days (P<0.05) with an inverse correlation to the neurological scores (r=-0.68, P=0.01). Immunohistochemistry detected the presence of Foxp3 primarily in the ischemic region of the brain tissue 1 day after cerebral ischemia; the contralateral hemisphere also showed a small quantity of Foxp3 expression. No Foxp3 expression was detected in the brain tissue of the sham-operated group. CONCLUSION: CD4(+)CD25 T regulatory cells participate in the inflammatory immune reactions as early as 1 day after acute cerebral ischemia in rats, which might be a protective mechanism of the brain cells.


Assuntos
Isquemia Encefálica/imunologia , Isquemia Encefálica/metabolismo , Fatores de Transcrição Forkhead/metabolismo , Linfócitos T Reguladores/metabolismo , Animais , Encéfalo/metabolismo , Fatores de Transcrição Forkhead/imunologia , Masculino , Ratos , Ratos Wistar , Linfócitos T Reguladores/imunologia
18.
Nan Fang Yi Ke Da Xue Xue Bao ; 31(10): 1792-4, 2011 Oct.
Artigo em Chinês | MEDLINE | ID: mdl-22027794

RESUMO

OBJECTIVE: To report a case of critical illness polyneuropathy (CIP) with Parkinson disease and discuss the development, clinical features and early diagnosis of this condition. METHODS: The clinical data of a patient with CIP and Parkinson's disease and the relevant literature were reviewed. RESULTS: This case showed no typical disease course of sepsis, and the condition exacerbated rapidly. The patient presented initially with abnormal homeostasis, followed by rapid onset of respiratory muscle weakness to require mechanical ventilation, but no limb weaknesses were detected. Intravenous antibiotics and aggressive treatment of sepsis did not produce any positive responses to wean from mechanical ventilation. Examinations of creatine kinase and cerebrospinal fluid showed no abnormalities. Electromyography and nerve conduction studies demonstrated declined nerve conduction velocity and decreased sensory and motor muscle action potentials, suggesting the possibility of CIP. CONCLUSION: In patients with Parkinson disease, the occurrence of sepsis with prolonged mechanical ventilation and limb weakness indicates the necessity of neurophysiological examination, muscle biopsies and laboratory tests, which may help detect CIP in the early phase. Proper interventions of sepsis may reduce the likeliness of CIP. Elimination of the risk factors and aggressive management of sepsis can be effective measures for preventing CIP.


Assuntos
Doença de Parkinson/complicações , Polineuropatias/complicações , Polineuropatias/diagnóstico , Sepse/complicações , Idoso , Humanos , Masculino , Respiração Artificial , Insuficiência Respiratória/complicações
19.
Cytotherapy ; 13(1): 46-53, 2011 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-20735164

RESUMO

BACKGROUND AIMS: This study aimed to observe nine factors expressed in rat ischemic brain after transplantation of bone marrow stromal cells (BMSC) and/or endothelial progenitor cells (EPC). These factors were vascular endothelial growth factor (VEGF), stromal cell-derived factor-1 (SDF-1), basic fibroblast growth factor (bFGF), insulin-like growth factor (IGF-l), transforming growth factor-ß (TGF-ß), platelet-derived growth factor-BB (PDGF-BB), brain-derived neurotrophic factor (BDNF), glial cell line-derived neurotrophic factor (GDNF) and nerve growth factor (NGF). METHODS: Adult Wistar rats were divided randomly into four groups: a vehicle group, BMSC group, EPC group and BMSC combined with EPC group. The rats were subjected to middle cerebral artery occlusion (MCAO) then implanted intravenously with 3 × 10(6) BMSC, EPC, BMSC/EPC or phosphate-buffered saline (PBS) 24 h after MCAO. Neurologic functional deficits were measured on days 1, 7, 14, 28 after transplantation. On day 7 after transplantation, quantitative reverse transcription (qRT)-polymerase chain reaction (PCR) and Western blot were employed to detect the expression of VEGF, SDF-1, bFGF, IGF-l, TGF-ß, PDGF-BB, BDNF, GDNF and NGF. RESULTS: The neurologic evaluation found that the neurologic severity scores were no different between the four groups on day 1, and the scores of rats in the BMSC/EPC group were significantly lower than those of rats in the other groups on days 7, 14 and 28 after transplantation. The expressions of bFGF, VEGF and BNDF were significantly higher in the BMSC/EPC group compared with the other groups. CONCLUSIONS: The intravenous transplantation of BMSC combined with EPC could promote the functional rehabilitation of rats with focal cerebral ischemia, and the mechanism may be related to the enhanced expression of factors.


Assuntos
Células da Medula Óssea/citologia , Isquemia Encefálica/metabolismo , Encéfalo/metabolismo , Encéfalo/patologia , Citocinas/metabolismo , Células Endoteliais/transplante , Transplante de Células-Tronco , Animais , Comportamento Animal , Células da Medula Óssea/metabolismo , Isquemia Encefálica/patologia , Isquemia Encefálica/fisiopatologia , Isquemia Encefálica/terapia , Células Endoteliais/citologia , Microvasos/patologia , Fatores de Crescimento Neural/genética , Fatores de Crescimento Neural/metabolismo , Ratos , Ratos Wistar , Células-Tronco/citologia , Células-Tronco/metabolismo , Células Estromais/citologia , Células Estromais/transplante
20.
Int J Cancer ; 127(9): 2222-9, 2010 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-20127864

RESUMO

Inhibition of tumor neovascularization has profound effects on the growth of solid tumors. Our previous studies have shown the effect of VEGF165-PE38 recombinant immunotoxin on proliferation and apoptosis in human umbilical vein endothelial cells in vitro. In this study, we explored the direct inhibition of angiogenesis in chick chorioallantoic membrane and antiangiogenic therapy in a malignant glioma model. HEK293 cells were transfected with the pVEGF165PE38-IRES2-EGFP plasmid. ELISA was used to confirm the expression of VEGF165-PE38 in the transfected cells. These cells released 1396 + or - 131.9 pg VEGF165-PE38/1x10(4) cells/48 h into the culture medium and the supernatant was capable of inhibiting the growth of capillary-like structures in chick chorioallantoic membrane assay. In a murine malignant glioma model, plasmid was directly administered via multiple local intratumoral delivery. After day 16 the tumor volume in mice treated with pVEGF165PE38-IRES2-EGFP was significantly lower than that in mice in the control groups. Immunohistochemistry studies showed that the treated group had decreased expression of CD31. Quantitative analysis of microvessel density in the treated group was 1.99 + or - 0.69/0.74 mm(2), and was significantly lower than that in the control groups (9.33 + or - 1.99/0.74 mm(2), 8.09 + or - 1.39/0.74 mm(2) and 8.49 + or - 1.69/0.74 mm(2)). Immunohistochemistry analysis indicated that immunotoxin VEGF165-PE38 was distributed in the treated group in malignant glioma tissue. Our findings provide evidence that the in vivo production of VEGF165-PE38 through gene therapy using a eukaryotic expression plasmid had potential antiangiogenic activity in malignant glioma in vivo.


Assuntos
Inibidores da Angiogênese/uso terapêutico , Terapia Genética , Glioma/terapia , Imunotoxinas/uso terapêutico , Fator A de Crescimento do Endotélio Vascular/genética , ADP Ribose Transferases/uso terapêutico , Animais , Toxinas Bacterianas/uso terapêutico , Linhagem Celular Tumoral , Exotoxinas/uso terapêutico , Estudos de Viabilidade , Glioma/irrigação sanguínea , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Plasmídeos , Pseudomonas/metabolismo , Transfecção , Fatores de Virulência/uso terapêutico , Ensaios Antitumorais Modelo de Xenoenxerto
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