RESUMO
Objective: To observe mutual interactions between macrophages(Mφ) and glioma stem cells (GSCs)in dual-color tracing model in vitro, to identify the biological characteristics of fusion cells in multiple levels, and to analysis the relevant molecular mechanisms. Methods: Red fluorescent protein(RFP) gene was stably transfected into human GSCs cell line SU4. Mφ cells were obtained from Balb/c nude mice with enhanced green fluorescent protein (EGFP) expression. Then two cells were co-cultured in dual-color tracing platform. RFP/EGFP double positive cells with high proliferation ability were mono-cloned. The fusion cells were verified by Western blot, fluorescence in situ hybridization, immunocytochemistry and chromosome karyotype analysis.The biological characteristics of fusion cells were further analyzed, together with relevant molecular changes. Results: RFP / EGFP double positive cells were obtained through in vitro co-culture. RFP and EGFP coexpression were proved at transcriptional and translational levels in the fusion cells. They also co-expressed GSCs marker Nestin and Mφ marker CD68, and karyotype analysis showed two types of characteristic chromosomes, which confirmed that the fusion cells originated from spontaneous fusion between SU4-RFP and Mφ.Fusion cell proliferation rate and invasion ability were higher than SU4-RFP, which were relevant with down-regulation of miR-146b-5p and activation of STAT3. Fusion cells transfected with miR-146b-5p showed a higher apoptosis rate(18.83%) and lower tumor formation(4/5). Conclusion: Mφ could fuse with GSCs spontaneously in local tumor micro-environment. The proliferation and invasion abilities of fusion cells were higher than their parent cells, which were relevant with down-regulation of miR-146b-5p and activation of STAT3. It revealed the possible mechanisms of malignant progression of gliomas.
Assuntos
Técnicas de Cocultura , Regulação para Baixo , Glioma , Macrófagos , Animais , Comunicação Celular , Fusão Celular , Linhagem Celular Tumoral , Transformação Celular Neoplásica , Proteínas de Fluorescência Verde , Humanos , Hibridização in Situ Fluorescente , Proteínas Luminescentes , Camundongos , Camundongos Nus , MicroRNAs , Células-Tronco Neoplásicas , Transfecção , Proteína Vermelha FluorescenteRESUMO
Despite the critical roles of excited states in protein functions, they remain intractable for most structural studies because of their notably low populations. Chemical shifts for "invisible" states in slow exchange with the ground state are intuitively observed using nuclear magnetic resonance (NMR) chemical exchange saturation transfer (CEST) experiments. Here, we present a CEST NMR spectroscopy study for the observation of protein pseudocontact shifts (PCSs) of excited states, which carry valuable angular and distance information about these states. We first validate this approach, dubbed PCS-CEST, in the slow-exchange system of Abp1p SH3-Ark1p labeled with lanthanide ions, where the PCSs of the minor states resemble those of the holo-form ground state as expected. We further demonstrate that pre-existing folding transitional conformations of an FF domain exhibit remarkably lower PCS values than the ground state, which suggests that the low-populated ensemble is unfolded or largely unfolded. A higher resolution of PCSs of the minor states is achieved using our 1D selective CEST experiments. Thus, PCS-CEST provides an exquisite structural probe into the minor but functionally essential excited states.
RESUMO
This study aims to explore the roles of somatic embryogenesis receptor-like kinase (SERK) in Malus hupehensis (Pingyi Tiancha). The full-length sequences of SERK1 in triploid Pingyi Tiancha (3n) and a tetraploid hybrid strain 33# (4n) were cloned, sequenced, and designated as MhSERK1 and MhdSERK1, respectively. Multiple alignments of amino acid sequences were conducted to identify similarity between MhSERK1 and MhdSERK1 and SERK sequences in other species, and a neighbor-joining phylogenetic tree was constructed to elucidate their phylogenetic relations. Expression levels of MhSERK1 and MhdSERK1 in different tissues and developmental stages were investigated using quantitative real-time PCR. The coding sequence lengths of MhSERK1 and MhdSERK1 were 1899 bp (encoding 632 amino acids) and 1881 bp (encoding 626 amino acids), respectively. Sequence analysis demonstrated that MhSERK1 and MhdSERK1 display high similarity to SERKs in other species, with a conserved intron/exon structure that is unique to members of the SERK family. Additionally, the phylogenetic tree showed that MhSERK1 and MhdSERK1 clustered with orange CitSERK (93%). Furthermore, MhSERK1 and MhdSERK1 were mainly expressed in the reproductive organs, in particular the ovary. Their expression levels were highest in young flowers and they differed among different tissues and organs. Our results suggest that MhSERK1 and MhdSERK1 are related to plant reproduction, and that MhSERK1 is related to apomixis in triploid Pingyi Tiancha.
Assuntos
Flores/genética , Malus/genética , Filogenia , Sequência de Aminoácidos/genética , Proteínas de Arabidopsis/genética , Clonagem Molecular , Flores/crescimento & desenvolvimento , Regulação da Expressão Gênica de Plantas , Malus/crescimento & desenvolvimento , Proteínas de Plantas/biossíntese , Proteínas de Plantas/genética , Proteínas Quinases/genética , Alinhamento de Sequência , Tetraploidia , TriploidiaRESUMO
OBJECTIVE: Acute myocardial infarction (AMI) is a serious cardiovascular disease with a high incidence worldwide and the main cause of sudden cardiac death. The aim of this article was to study the protective role of miR-335 in myocardial infarction (MI) and the underlying molecular mechanism. MATERIALS AND METHODS: Thirty Sprague Dawley (SD) rats were randomly divided into sham group, MI + NC group and MI + agomiR-335 group. The expression of miR-335 in rat myocardium was detected by quantitative Real Time-Polymerase Chain Reaction (RT-PCR). Western blot was performed to detect the expression of TNF-α, IL-6, IL-1ß, Caspase-3, Cleaved Caspase-3 (C-Caspase-3) and MAP3K2 in rat myocardium. On the 7th day of the establishment of the rat MI model, a high-resolution small animal ultrasound system was utilized to detect the cardiac function of the rats, and the heart tissues and blood samples of the rats were collected. The corresponding kits were purchased to detect the contents of LDH, CK-MB, MDA and SOD in rat serum, and HE staining was employed to observe the morphology of rat myocardial tissue. RESULTS: The expression of miR-335 in myocardial infarcted zones and border zones of MI rats decreased significantly. The upregulation of miR-335 remarkably inhibited myocardial inflammation and apoptosis after MI, thus improving the cardiac function of MI rats. Compared with the sham group, the MAP3K2 expression in the MI + NC group was observably increased, while the overexpression of miR-335 could inhibit the expression of this protein. Through the Luciferase reporter gene experiment, we found that miR-335 could directly target MAP3K2. CONCLUSIONS: The expression of miR-335 decreased in myocardial tissue after MI, and the overexpression of miR-335 reduced myocardial damage by inhibiting oxidative stress, inflammation, and apoptosis via targeting MAP3K2, thereby improving the cardiac function of MI rats.
Assuntos
MAP Quinase Quinase Quinase 2/metabolismo , MicroRNAs/metabolismo , Infarto do Miocárdio/metabolismo , Miócitos Cardíacos/metabolismo , Regulação para Cima , Animais , Ecocardiografia , Feminino , MicroRNAs/genética , Infarto do Miocárdio/patologia , Miócitos Cardíacos/patologia , Ratos , Ratos Sprague-DawleyRESUMO
In 14 ventilated, normocapnic baboons anaesthetised with alpha-chloralose, local CBF (hydrogen clearance) and the amplitude and latency of local components of the somatosensory evoked potential (SEP, median nerve stimulation) were measured bilaterally in ventrobasal thalamus (VPL), medial lemniscus (ML), and cerebral cortex before and during progressive ischaemia, produced by occlusion of the right middle cerebral artery and subsequent controlled reductions in mean systemic blood pressure (MSBP). The first significant reduction from control of the left cortical SEP amplitude occurred in the range of 30-40 mm Hg MSBP, but those of the VPL and ML responses only below 30 mm Hg; in the range of 20-30 mm Hg, the average SEP amplitudes in cortex, VPL, and ML were 8.6, 72.6, and 90.7% of control, respectively. In terms of local CBF, the cortical SEP threshold was in the range of 15-20 ml/100 g/min (as in previous work), that of VPL in the range of 10-15 ml/100 g/min, but the ML response was only markedly reduced below 10 ml/100 g/min. Thus, the differential ischaemic sensitivity of the SEP between the three regions was clearly demonstrated. These results indicate that as one descends the neuraxis, there is an increasing resistance of electrophysiological function to systemic hypotension, together with a decreasing threshold for local ischaemia.
Assuntos
Isquemia Encefálica/fisiopatologia , Tronco Encefálico/fisiopatologia , Córtex Cerebral/fisiopatologia , Potenciais Somatossensoriais Evocados , Tálamo/fisiopatologia , Animais , Arteriopatias Oclusivas/complicações , Arteriopatias Oclusivas/fisiopatologia , Isquemia Encefálica/etiologia , Circulação Cerebrovascular , Papio , Tempo de ReaçãoRESUMO
Carotid-cavernous sinus fistulas are not rare, but they have never been reported in association with persistent primitive trigeminal artery. We recently encountered such a case. The Jaeger-Hamby procedure was employed, with mandatory occlusion of the primitive trigeminal artery.
Assuntos
Fístula Arteriovenosa/cirurgia , Doenças das Artérias Carótidas/cirurgia , Seio Cavernoso , Nervo Trigêmeo/irrigação sanguínea , Adulto , Artérias/anormalidades , Artérias/cirurgia , Fístula Arteriovenosa/etiologia , Doenças das Artérias Carótidas/etiologia , Artéria Carótida Interna , Traumatismos Craniocerebrais/complicações , Humanos , Recém-Nascido , MasculinoRESUMO
The authors report a case of acoustic neurinoma presenting as intratumoral bleeding. This is the tenth reported occurrence. The literature is reviewed. All cases that have been reported have appeared with sudden onset of headache, vomiting, and decreased levels of consciousness. Aneurysm rupture, an arteriovenous malformation, or other vascular anomalies are suspected first. Preexisting unilateral hearing impairment is a valuable clue to differential diagnosis. Contrast-enhanced computed tomographic scans and cerebral angiograms are important tools for correct diagnosis. The tumor size (greater than 2 cm) and the thin, dilated vessels within the tumor are considered as pathogenetic factors for bleeding. When neurological status is not stable, placement of a ventriculoperitoneal shunt followed by urgent extirpation of the tumor is indicated.
Assuntos
Neuroma Acústico/irrigação sanguínea , Idoso , Feminino , Hemorragia/patologia , Humanos , Neuroma Acústico/diagnóstico por imagem , Neuroma Acústico/patologia , Tomografia Computadorizada por Raios XRESUMO
The relationship between central conduction time (CCT) and hemispheric cerebral blood flow (CBF) has been examined in 20 patients presenting with subarachnoid hemorrhage. A total of 63 combined CCT/CBF recordings were performed at various times throughout the hospital course of these patients, and the findings were correlated to clinical status. The initial-slope index of the CBF (CBF isi) was found to correlate well with clinical grade, and a gradation in flow was noted between the different neurological grades. Patients in Grades I and II (Hunt and Hess classification) had the highest flows (mean CBF isi = 47.2 +/- 8.1); Grade III patients had intermediate flows (mean CBF isi = 39.6 +/- 7.8); and Grade IV patients had the lowest flows (mean CBF isi = 32.0 +/- 6.4). While CCT tended to become increasingly prolonged with worsening grade, a significant difference could not be demonstrated between Grade I, II, and III patients. Only when Grade IV status was reached was the CCT significantly prolonged. When CBF isi and CCT were examined, a threshold relationship was noted between CBF isi and CCT prolongation. At flow values above 30, little change was noted in CCT, and CCT remained in the normal range. However, at flow values below 30, CCT became increasingly prolonged as blood flow diminished. The degree of CCT prolongation appeared to be directly proportional to the degree of blood flow diminution at flows below threshold.
Assuntos
Circulação Cerebrovascular , Potenciais Somatossensoriais Evocados , Aneurisma Intracraniano/fisiopatologia , Condução Nervosa , Hemorragia Subaracnóidea/fisiopatologia , Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Vias Neurais/fisiologia , Córtex Somatossensorial/fisiologia , Medula Espinal/fisiologia , Transmissão SinápticaRESUMO
Somatosensory evoked responses to median nerve stimulation have been recorded during the management of subarachnoid hemorrhage from intracranial aneurysms. The type of measurement was central conduction time (CCT), the time interval between the N14 peak (at C-2) and the N20 peak (at the cortex). Significant differences were found between patients with aneurysms in clinical Grade 4 and normal individuals, although there was no significant difference between most aneurysm patients in Grades 1 through 3. The prolongation of CCT was found of prognostic value. Significant prolongation (mean plus two standard deviations) from the normal period of 5.4 +/- 0.4 msec was found significantly useful to predict a less than satisfactory outcome (a patient with neurological signs or disabled) at 2 months, and these differences were detectable as early as 48 hours postoperatively. Differences between conduction time in the two hemispheres could also be used prognostically, although the significance of the differences did not appear until 48 to 72 hours after surgery, and was in any event less than that of CCT.
Assuntos
Potenciais Somatossensoriais Evocados , Hemorragia Subaracnóidea/cirurgia , Adulto , Idoso , Encéfalo/fisiopatologia , Feminino , Humanos , Aneurisma Intracraniano/complicações , Aneurisma Intracraniano/cirurgia , Masculino , Pessoa de Meia-Idade , Condução Nervosa , Hemorragia Subaracnóidea/etiologia , Hemorragia Subaracnóidea/fisiopatologiaRESUMO
Somatosensory evoked potentials have been recorded during 34 operations for intracranial aneurysm. The central conduction time (CCT), the time between the N14 peak (recorded at C-2) and the N20 peak (recorded at the cortex) in response to median nerve stimulation, has been found to be increased by administration of halothane, by brain retraction, and by temporary vascular occlusion in some instances. Increase of CCT to more than 10 msec, or disappearance of the response was associated in two cases with postoperative neurological deficit, neither permanent. In a further case, prolongation of CCT was used as a guide to the repositioning of an imperfectly placed clip on a middle cerebral artery aneurysm. The value of the technique as a perioperative monitoring system is discussed.
Assuntos
Potenciais Somatossensoriais Evocados , Aneurisma Intracraniano/cirurgia , Adolescente , Adulto , Idoso , Encéfalo/fisiopatologia , Criança , Feminino , Humanos , Aneurisma Intracraniano/fisiopatologia , Cuidados Intraoperatórios , Masculino , Pessoa de Meia-Idade , Condução NervosaRESUMO
Infestation of the central nervous system with sparganum, the larva of Spirometra mansoni, is rare. Only 27 cases have been reported in the literature; however, the true incidence might be underestimated. Two cases are reported that were definitively diagnosed by stereotactic biopsy techniques. Without a positive histological diagnosis, the first case would have been diagnosed as having a metastatic brain tumor and a course of radiotherapy would probably have been initiated. Differentiation between granuloma of cerebral sparganosis and brain tumors such as meningiomas, gliomas, and metastatic tumors is often difficult before operation. Detailed magnetic resonance imaging might offer some help in such cases. Emphasis is placed on the increasing importance of stereotactic surgery in the diagnosis and treatment of an intracerebral mass lesion, including cerebral sparganosis.
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Encefalopatias/parasitologia , Encefalopatias/cirurgia , Esparganose/diagnóstico , Esparganose/cirurgia , Técnicas Estereotáxicas , Doenças dos Trabalhadores Agrícolas/parasitologia , Biópsia/métodos , Diagnóstico Diferencial , Feminino , Humanos , Pessoa de Meia-IdadeRESUMO
Flash visual evoked potentials (F-VEP's) have been recorded in fifteen patients (thirty eyes) with anterior visual pathway compression lesions before, during and after the operation in relation to clinical visual testing. A grading system for the wide variety of abnormal F-VEP's has been established, based on changes in forty patients (eighty eyes). No significant correlation between change in F-VEP and visual acuity or visual field has been demonstrated. Intraoperative F-VEP's have been found to be sensitive to halothane and to surgical manipulation of the optic nerve and/or chiasm during dissection of the tumour. The technique is considered a useful means of identifying the optic pathway during operation and may assist in prevention of optic damage if the transphenoidal approach is used.
Assuntos
Neoplasias Encefálicas/cirurgia , Craniofaringioma/cirurgia , Potenciais Evocados Visuais , Meningioma/cirurgia , Neoplasias Hipofisárias/cirurgia , Vias Visuais/fisiopatologia , Adulto , Neoplasias Encefálicas/fisiopatologia , Craniofaringioma/fisiopatologia , Feminino , Humanos , Período Intraoperatório , Masculino , Meningioma/fisiopatologia , Pessoa de Meia-Idade , Neoplasias Hipofisárias/fisiopatologia , Tempo de Reação/fisiologia , Acuidade Visual , Campos VisuaisRESUMO
Intraoperative use of somatosensory evoked potentials (SEP's) to monitor intracranial aneurysm surgery and flash visual evoked potentials (F-VEP's) for parasellar surgery have been routinely employed in our clinic. We found that both EP modalities are sensitive to the changing concentration of our standard hypotensive agent, halothane. The prolongation of the N14-N20 interpeak latency to median nerve stimulation at the wrist, and prolongation of P100 latency with altered configuration of early VEP components to flash light stimulation, appear to be the results of direct pharmacological effects of the agent and not an effect of secondary hypotension. VEP is found easily abolished by halothane at a concentration of 2.0%, while the SEP is more resistant. Halothane is not ideal however when monitoring intraoperative VEP.
Assuntos
Anestesia por Inalação , Sistema Nervoso Central/efeitos dos fármacos , Potenciais Somatossensoriais Evocados/efeitos dos fármacos , Potenciais Evocados Visuais/efeitos dos fármacos , Halotano/farmacologia , Adolescente , Adulto , Vias Aferentes/efeitos dos fármacos , Pressão Sanguínea , Encefalopatias/cirurgia , Criança , Feminino , Humanos , Aneurisma Intracraniano/cirurgia , Período Intraoperatório , Masculino , Pessoa de Meia-Idade , Vias Visuais/efeitos dos fármacosRESUMO
Changes in the central conduction time (CCT) during the application of temporary clips were studied in 40 patients who had undergone operations for intracranial aneurysms in relation to postoperative neurological outcome. Ten of these 40 patients (25%) showed postoperative morbidity, although promptly recoverable in 5. None of the patients whose CCTs did not change following temporary occlusion of major vessels showed any postoperative morbidity, except in one case of anterior cerebral artery aneurysm. In 6 patients, temporary vascular occlusion caused a considerable transient prolongation in CCT of up to 10 msec. Two of these 6 patients were associated with postoperative neurological deficit (which was recoverable in 1). The cortical response became flat in 15 patients. Seven of these 15 patients showed hemispheric deficits postoperatively, although recoverable in 4. There was a correlation between the change in the somatosensory evoked response and postoperative outcome. Disappearance of the N20 potential following occlusion is regarded as a danger signal, but postoperative, irrecoverable neurological deficit seems to be unlikely if its disappearance takes more than 3-4 minutes. Even if the cortical response disappears, the clinical outcome is expected to be good if the N20 potential recovers within 20 minutes after recirculation.
Assuntos
Potenciais Somatossensoriais Evocados , Aneurisma Intracraniano/cirurgia , Adulto , Idoso , Artéria Basilar , Artéria Carótida Interna , Artérias Cerebrais , Constrição , Feminino , Humanos , Período Intraoperatório , Masculino , Nervo Mediano/fisiologia , Pessoa de Meia-Idade , Condução Nervosa , PrognósticoRESUMO
Three cases of isolated spinal extradural cavernous hemangiomas are reported, two in the thoracic and one in the lumbar region. One of them manifested as acute cord compression, the other two as chronic progressive myelopathy or radiculopathy. A total excision or subtotal excision with irradiation was performed. All of them had good functional recovery. The clinical picture, radiological diagnosis, and optimal method of treatment are discussed. The relevant literature is reviewed.
Assuntos
Neoplasias Epidurais , Hemangioma Cavernoso , Adulto , Terapia Combinada , Diagnóstico Diferencial , Neoplasias Epidurais/complicações , Neoplasias Epidurais/diagnóstico , Neoplasias Epidurais/patologia , Neoplasias Epidurais/terapia , Feminino , Hemangioma Cavernoso/complicações , Hemangioma Cavernoso/diagnóstico , Hemangioma Cavernoso/patologia , Hemangioma Cavernoso/terapia , Humanos , Laminectomia , Masculino , Pessoa de Meia-Idade , Mielografia , Tomografia Computadorizada por Raios XRESUMO
Intramedullary spinal cord metastasis (ISM) is an unusual cause of myelopathy, in comparison to the common problem of extramedullary metastasis. Most patients with ISM are found at autopsy. When patients with ISM are diagnosed premorbidly, they usually have a previously known or concurrently diagnosable primary neoplasm. Radiation therapy is the definitive therapy. We recently encountered a patient who was seen initially because of progressive myelopathy, without evidence of underlying malignancy, in whom a papillary carcinoma was found at myelotomy. The condition improved after surgery and the patient returned to his previous work for five months. Subsequently, a lesion histologically identical with that obtained from the previous myelotomy was found in the lung. This case illustrates the value of surgical intervention in selected situation. When there is no identifiable organ metastasis or multiple intramedullary lesions, radical microsurgical resection of a focal intramedullary lesion should be considered.
Assuntos
Carcinoma Papilar/secundário , Neoplasias da Medula Espinal/secundário , Carcinoma Papilar/cirurgia , Humanos , Masculino , Pessoa de Meia-Idade , Neoplasias da Medula Espinal/cirurgiaRESUMO
A patient who noted the onset of repetitive arterial epistaxis and progressive unilateral visual impairment 2 months after a blunt head injury was found to have a traumatic aneurysm of the cavernous portion of internal carotid artery (ICA). The epistaxis resolved and his vision recovered following a trapping procedure. This case and a review of previously reported cases emphasize the importance of early cerebral angiography in patients with post-traumatic epistaxis and visual failure.
Assuntos
Artéria Carótida Interna/diagnóstico por imagem , Traumatismos Craniocerebrais/complicações , Epistaxe/etiologia , Aneurisma Intracraniano/complicações , Adulto , Lesões das Artérias Carótidas , Artéria Carótida Interna/cirurgia , Epistaxe/fisiopatologia , Humanos , Aneurisma Intracraniano/etiologia , Aneurisma Intracraniano/cirurgia , Masculino , Tomografia Computadorizada por Raios X , Acuidade VisualRESUMO
A series of 52 patients suffering post-traumatic basal ganglia hemorrhage (BGH) after closed head injury is reviewed. Post-traumatic BGH was associated with other intracranial lesions in most cases. The outcome in this series is rather good, with 53.8% of patients having a functional survival. Radiologically, the post-traumatic intraventricular hemorrhage (IVH) and brainstem hemorrhage usually coexisted with a poor outcome. Old age (greater than 60), abnormal pupil reaction, impaired oculocephalic response, and abnormal motor response to pain stimuli are reliable clinical hallmarks for poor prognosis.
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Gânglios da Base/lesões , Hemorragia Cerebral/etiologia , Traumatismos Craniocerebrais/complicações , Adolescente , Adulto , Fatores Etários , Idoso , Hemorragia Cerebral/mortalidade , Hemorragia Cerebral/fisiopatologia , Criança , Pré-Escolar , Coma/etiologia , Traumatismos Craniocerebrais/patologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Exame Neurológico , Paralisia/etiologia , Prognóstico , Estudos RetrospectivosRESUMO
A relatively rare case of cerebellar arteriovenous malformation (AVM) associated with a saccular aneurysm fed by the left posterior inferior cerebellar artery (PICA) is described. The interpretation of angiograms and the appropriate surgical treatment for the coexistence of the intracranial AVM and aneurysm are discussed. It is a fact that the steal phenomenon existed in the flow of contrast medium during the angiography. The appropriate surgical treatment may be clipping of the aneurysmal neck first and then radical excision of the AVM.