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Objective:To explore the effect of task-based rehabilitative training on neural circuit plasticity and forelimb motor function after C5 spinal cord injury in mice. Methods:A total of 21 healthy C57/BL mice were randomly and equally divided into sham group, model group and training group. The model was established by left C5 spinal cord crush injury. The lamina was removed without damaging the spinal cord in the sham group. Four weeks after injury, the training group received task-based rehabilitative training for four weeks. The horizontal ladder and rearing tests were used to assess motor function for forelimb before injury, and three days, two weeks, four weeks, six weeks and eight weeks after injury. The axons of the corticospinal tract in all mice were observed six weeks after injury by using biotinylated dextran amin (BDA) anterograde tracing. Eight weeks after injury, motor-evoked potential was applied to measure nerve conduction velocities in forelimb, while the axon sprouting and syntagmatic relation of neuron in the anterior horn of gray matter above lesion were observed by immunofluorescence double-labeling of BDA/neuron-specific nuclei protein (NeuN); the expression of Synapsin in the anterior horn of gray matter at lesion was observed by immunofluorescence double-labeling of NeuN/Synapsin I. Results:Eight weeks after injury, the latency of P1 and N1 was longer in the model group than in the sham group (P < 0.05), and was shorter in the training group than in the model group (P < 0.05). Compared with the sham group, the error rate of left forelimb increased, and the usage rate decreased (P < 0.05) in the model group and the training group; compared with the model group, the error rate of left forelimb decreased six weeks and eight weeks after injury (P < 0.05), and the usage rate increased eight weeks after injury (P < 0.05) in the treatment group. Compared with the model group, more axon sprouting co-localized with neurons in the anterior horn of gray matter above lesion (P < 0.05), and the expression of Synapsin I increased in the training group (P < 0.05). Conclusion:Task-based rehabilitative training could promote the neural circuit plasticity and improve the motor function of forelimb after spinal cord injury in mice.
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Rehabilitation is an essential treatment for knee osteoarthritis. Rehabilitation evaluation is the foundation of rehabilitation treatment. The items of rehabilitation evaluation for knee osteoarthritis include pain, proprioception, range of motion, muscle force, balance, gait, quality of life, and so on. When evaluating the patient's pain, researchers should not only focus on Visual Analogue Score (VAS) outcomes, but also consider the patient's body function and activity level, and whether there is neuropathic pain. Furthermore, isokinetic dynamometer can not only be used for quantitative assessment for muscle strength, but also for the quantitative evaluation for patient's proprioception. Moreover, Community Balance and Mobility Scale has no ceiling and floor effects which also is a reliable and useful tool for assessing the balance function in patients with knee osteoarthritis, compared with Berg Balance Scale. The evaluation for body structure in patients with knee osteoarthritis mainly includes the radiological examination and physical examination. There are a lot of scales to evaluate the quality of life in patients with knee osteoarthritis, and Osteoarthritis of Knee and Hip Quality of Life fits most. Further researches and promotion of evaluation tools for knee osteoarthritis are needed in the future.
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Objective To observe the marker effects of two different fluorescent dyes, DIL and DAPI, in labeling endogenous neural stem cells (ENSCs) in rat central nervous system. Methods Thirty-six Sprague-Dawley rats were randomized into staining groups, comprising DIL group and DAPI group, and the corresponding control groups, including DMSO group for DIL group and PBS group for DAPI group. 0.2% DIL 10μl or 10μg/ml DAPI 10μl was stereotactically injected into the lateral ventricle of rats of DIL group or DAPI group, while DMSO or PBS 10μl was introduced into that of DMSO group or PBS group. Neurological severity score (NSS) was determined 2 hours and 24 hours respectively after the operation. Rats were sacrificed at day 1, 3, 7 after the injection. Serial coronal sections of the brain and spinal cord were carried out on a cryostat, and then they were observed under a confocal microscope. The fluorescence intensity of the targeted area, which highlighted by labeled ependymal cells, in the brain and spinal cord of cervical vertebrae, thoracic vertebrae and lumbar vertebrae were semi-quantified. Fluorescence intensity of each section was measured in triplicate, and a mean value was obtained. Statistical analysis was performed on 3 data sets, randomly selected from sections of brain and spinal cord obtained at day 1, 3, 7. Results Two hours after DIL injection, the rats showed no evident neurological defect. NSS value was very low, and there was no significant difference compared with the DMSO group (P>0.05). Twenty-four hours later, normal neurological function recovered in all the rats. Red fluorescence could be seen in the cytoplasm of ependymal cells in the lateral ventricle and each spinal cord segment at day 1 after the DIL injection, and it did not disappear until the 7th day. Nuclei of DAPI-labeled lateral ventricle cells were blue, with clear nuclear morphology. Choroid plexus cells of the ventricle were also labeled. However, there was no blue fluorescence in the medulla oblongata or any segment ofthe spinal cord. The picture at day 3 and day 7 was similar to that of day 1. No significant difference was found between fluorescence intensity in DIL or DAPI stained cells(P>0.05) at any time point. Conclusions DIL may serve as a marker of the cytoplasm of ependymal cells in the brain ventricle and spinal central canal. DAPI, which is often used in the nuclear staining, can label the ENSCs in the brain ventricle. Intraventricular injection of fluorescent dye is a relatively safety procedure.
RESUMO
Objective To observe the marker effects of two different fluorescent dyes, DIL and DAPI, in labeling endogenous neural stem cells (ENSCs) in rat central nervous system. Methods Thirty-six Sprague-Dawley rats were randomized into staining groups, comprising DIL group and DAPI group, and the corresponding control groups, including DMSO group for DIL group and PBS group for DAPI group. 0.2% DIL 10μl or 10μg/ml DAPI 10μl was stereotactically injected into the lateral ventricle of rats of DIL group or DAPI group, while DMSO or PBS 10μl was introduced into that of DMSO group or PBS group. Neurological severity score (NSS) was determined 2 hours and 24 hours respectively after the operation. Rats were sacrificed at day 1, 3, 7 after the injection. Serial coronal sections of the brain and spinal cord were carried out on a cryostat, and then they were observed under a confocal microscope. The fluorescence intensity of the targeted area, which highlighted by labeled ependymal cells, in the brain and spinal cord of cervical vertebrae, thoracic vertebrae and lumbar vertebrae were semi-quantified. Fluorescence intensity of each section was measured in triplicate, and a mean value was obtained. Statistical analysis was performed on 3 data sets, randomly selected from sections of brain and spinal cord obtained at day 1, 3, 7. Results Two hours after DIL injection, the rats showed no evident neurological defect. NSS value was very low, and there was no significant difference compared with the DMSO group (P>0.05). Twenty-four hours later, normal neurological function recovered in all the rats. Red fluorescence could be seen in the cytoplasm of ependymal cells in the lateral ventricle and each spinal cord segment at day 1 after the DIL injection, and it did not disappear until the 7th day. Nuclei of DAPI-labeled lateral ventricle cells were blue, with clear nuclear morphology. Choroid plexus cells of the ventricle were also labeled. However, there was no blue fluorescence in the medulla oblongata or any segment ofthe spinal cord. The picture at day 3 and day 7 was similar to that of day 1. No significant difference was found between fluorescence intensity in DIL or DAPI stained cells(P>0.05) at any time point. Conclusions DIL may serve as a marker of the cytoplasm of ependymal cells in the brain ventricle and spinal central canal. DAPI, which is often used in the nuclear staining, can label the ENSCs in the brain ventricle. Intraventricular injection of fluorescent dye is a relatively safety procedure.