Functional electronic stimulation (FES) may provide a means to restore motor function in patients with spinal cord injuries. The goal of this study is to determine the regions in the spinal cord controlling different hindlimb movements in the rats. Normalization was used for the regions dominating the corresponding movements. It has been verified that FES can be used in motor function recovery of the hindlimb. The spinal cord was stimulated by FES with a three-dimensional scan mode in experiments. The results show that stimulation through the electrodes implanted in the ventral locations of the lumbosacral enlargement can produce coordinated single- and multi-joint hindlimb movements. A variety of different hindlimb movements can be induced with the appropriate stimulation sites, and movement vectors of the hindlimb cover the full range of movement directions in the sagittal plane of the hindlimb. This article drew a map about spinal cord motor function of the rat. The regions in the spinal cord which control corresponding movements are normalized. The data in the study provide guidance about the location of electrode tips in the follow-up experiments.
Objective To explore the effects of human urine-derived stem cells (hUSCs) and hUSCs combined with chondroitinase ABC (chABC) on the expressions of nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF) in the spinal cord injury (SCI) of rats, and to investigate the underlying mechanism. Methods hUSCs were cultured from human urine, and their phenotypes were detected by flow cytometry. The SCI model of rats were made via Allen method. Sixty Sprague Dawley rats were divided into 5 groups (n=12): the sham operation group (group A), SCI group (group B), SCI+hUSCs group (group C), SCI+chABC group (group D), and SCI+hUSCs+chABC group (group E). Basso, Beattie, Bresnahan (BBB) score was used to measure the lower extremity motor function of rats in each group at 10, 20, and 30 days after operation. Real-time fluorescent quantitative PCR was used to detect the relative mRNA expressions of NGF and BDNF at 30 days. Meanwhile, the protein expression of NGF and BDNF were confirmed by immunohistochemistry staining. The relative protein expressions of Bax and Bcl-2 were detected by Western blot. Results The hUSCs were identified to have multipotential differentiation potential. At 10, 20, and 30 days, BBB score was significantly lower in group B than in groups A, C, D, and E, in groups C, D, and E than in group A, in groups C and D than in group E (P<0.05). Real-time fluorescent quantitative PCR and immunohistochemistry staining demonstrated that the expressions of NGF and BDNF were significantly lower in group B than in groups A, C, D, and E, in groups C, D, and E than in group A, in groups C and D than in group E (P<0.05); but there was no significant difference between groups C and D (P>0.05). Western blot results indicated that the protein expression of Bax was significantly higher in group B than in groups A, C, D, and E, in groups C, D, and E than in group A, in groups C and D than in group E (P<0.05). Meanwhile, the protein expression of Bcl-2 was significantly lower in group B than in groups A, C, D, and E, in groups C, D, and E than in group A, in groups C and D than in group E (P<0.05). Conclusion hUSCs can protect SCI and this positive effect can be enhanced by chABC; this neuro-protective effect may depend on promoting the expressions of NGF and BDNF, and suppressing the neuronal apoptosis.
ObjectiveTo investigate the expression changes and the repair effect of mitogen and stress- activated protein kinase 1 (MSK1) on spinal cord injury (SCI) in rats.MethodsOne hundred and twenty male Sprague Dawley (SD) rats (weighing 220-250 g) were used for the study, 70 of them were randomly divided into sham-operation group and SCI group (n=35), the rats in SCI group were given SCI according to Allen’s method, and the sham-operation group only opened the lamina without injuring the spinal cord; spinal cord tissue was collected at 8 hours, 12 hours, 1 day, 2 days, 3 days, 5 days, and 7 days after invasive treatment, each group of 5 rats was used to detect the expression of MSK1 and proliferating cell nuclear antigen (PCNA) by Western blot assay. Another 20 SD rats were grouped by the same method as above (n=10). In these rats, a negative control lentiviral LV3NC dilution was injected at a depth of approximately 0.8 mm at the spinal cord T10 level. The results of transfection at 1, 3, 5, 7, and 14 days after injection were observed under an inverted fluorescence microscope to determine the optimal transfection time of the virus. The other 30 SD rats were randomly divided into group A with only SCI, group B with a negative control lentiviral LV3NC injected after SCI, and group C with MSK1 small interfering RNA (siRNA) lentivirus injected after SCI, with 10 rats each group. The Basso, Beatlie, Bresnahan (BBB) score of hind limbs was measured at 1, 3, 5, 7, and 14 days after treatment; spinal cord tissue collected at the optimal time point for lentivirus transfection was detected the expression changes of MSK1 and PCNA by Western blot and the localization by immunofluorescence staining of MSK1 and PCNA proteins.ResultsWestern blot assay showed that there was no significant changes in the expression of MSK1 and PCNA at each time points in the sham-operation group. In the SCI group, the expression of MSK1 protein was gradually decreased from 8 hours after injury to the lowest level at 3 days after injury, and then gradually increased; the expression change of PCNA protein was opposite to MSK1. The expression of MSK1 in SCI group was significantly lower than that in the sham-operation group at 1, 2, 3, and 5 days after injury (P<0.05), and the expression of PCNA protein of SCI group was significantly higher than that of the sham-operation group at 8 hours and 1, 2, 3, 5, and 7 days after injury (P<0.05). The fluorescence expression of both the SCI group and the sham-operation group has be found and peaked at 7 days. There was a positive correlation between fluorescence intensity and time in 7 days after transfection. With the prolongation of postoperative time, the BBB scores of groups A, B, and C showed a gradually increasing trend. The BBB score of group C was significantly lower than those of groups A and B at 5, 7, and 14 days after treatment (P<0.05). After transfection for 7 days, Western blot results showed that the relative expression of MSK1 protein in group C was significantly lower than that in groups A and B (P<0.05); and the relative expression of PCNA protein was significantly higher than that in groups A and B (P<0.05). Immunofluorescence staining showed that MSK1 was expressed in the nuclei of the spinal cord and colocalized with green fluorescent protein, neuronal nuclei, and glial fibrillary acidic protein (GFAP). The relative expression area of MSK1 positive cells in group C was significantly higher than that in group B (P<0.05), and the relative expression areas of PCNA and GFAP positive cells were significantly lower than those in group B (P<0.05).ConclusionLentivirus-mediated MSK1 siRNA can effectively silence the expression of MSK1 in rat spinal cord tissue. MSK1 may play a critical role in the repair of SCI in rats by regulating the proliferation of glial cells.
ObjectiveTo explore the feasibility and mechanism of inhibiting miR-429 to improve the permeability of the blood spinal cord barrier (BSCB) in vitro, and provide a new gene therapy target for enhancing the spinal cord microenvironment.MethodsFirst, the immortalized human brain microvascular endothelial cell line (hCMEC/D3) was transfected with the anti-miR-429 antagonist (antagomiR-429) and its negative control (antagomiR-429-NC), respectively. The miR-429 expression of hCMEC/D3 cells was observed by fluorescence microscopy and real-time fluorescence quantitative PCR to verify the transfection efficiency of antagomiR-429. Then the effect of miR-429 on BSCB permeability was observed in vitro. The experiment was divided into 4 groups. The blank control group (group A) was constructed of normal hCMEC/D3 cells and Ha-sc cells to prepare the BSCB model, the hypoxia-induced group (group B), the hypoxia-induced+antagomiR-429-NC group (group C), and the hypoxia-induced+antagomiR-429 group (group D) were constructed of normal, antagomiR-429-NC transfected, and antagomiR-429 transfected hCMEC/D3 cells and Ha-sc cells to prepare the BSCB models and hypoxia treatment for 12 hours. The permeability of BSCB in vitro was measured by horseradish peroxidase (HRP) permeability. Real-time fluorescence quantitative PCR, Western blot, and immunofluorescence staining were used to observe the expressions of ZO-1, Occludin, and Claudin-5.ResultsThe antagomiR-429 and antagomiR-429-NC were successfully transfected into hCMEC/D3 cells under a fluorescence microscope, and the transfection efficiency was about 90%. Real-time fluorescence quantitative PCR results showed that the relative expression of miR-429 in antagomiR-429 group was 0.109±0.013, which was significantly lower than that of antagomiR-429-NC group (0.956±0.004, P<0.05). HRP permeability measurement, real-time fluorescence quantitative PCR, and Western blot results showed that the HRP permeability of groups B and C were significantly higher than those of groups A and D (P<0.05), and the relative expressions of ZO-1, Occludin, and Claudin-5 proteins and mRNAs were significantly lower in groups B and C than in groups A and D (P<0.05) and in group D than in group A (P<0.05); there was no significant difference between groups B and C (P>0.05). Immunofluorescence staining showed that the immunofluorescence of ZO-1, Occudin, and Claudin-5 at the cell membrane boundary in group D were stronger than those in groups B and C, but not as strong as that in group A.ConclusionInhibition of miR-429 expression can promote the expressions of ZO-1, Occludin, and Claudin-5 proteins in microvascular endothelial cells, thereby improving the increased permeability of BSCB due to hypoxia.
Objective To investigate the effects of 17β-estradiol on the cell apoptosis after chronic spinal cord injury in ovariectomized rats. Methods A total of 90 female Wistar rats (weighing, 220-250 g) received removal of bilateral ovaries. After 2 weeks, the rats were randomly divided into 3 groups (n=30): sham-operation group (group A); chronic gradual spinal cord injury model and 17β-estradiol treatment group (group B); and chronic gradual spinal cord injury model and normal saline treatment group (group C). Rats of group A only received removal of spinous process at T10. Rats of groups B and C were made the models of chronic gradual spinal cord injury, and then 17β-estradiol (100 μg/kg, twice a week) and normal saline were given by peritoneal injection, respectively. The cell apoptosis and positive cells of Caspase-3 were examined by the TUNEL methods and Caspase-3 immunohistochemical staining at 1, 3, 7, 14, 28, and 60 days after modeling; and the neurological function was evaluated by Tarlov scale and inclined plane test scoring. Results At 14, 28, and 60 days after modeling, Tarlov scale and inclined plane test scores of group B were significantly better than those of group C (P lt; 0.05), but were significantly lower than those of group A (P lt; 0.05). At 28 days after modeling, HE staining showed that the edema of spinal gray matter and the neurons, the proliferation of glial cells and astrocytes, and less pathologic change were observed in group B; and the pathological changes in group B were mitigated than in group C. At 60 days after modeling, edema of spinal gray matter and the neurons was significantly ameliorated in group B. At 14, 28, and 60 days after modeling, the rate of Caspase-3 positive cells in group B was significantly lower than in group C (P lt; 0.05), but was significantly higher than in group A (P lt; 0.05). At 7, 14, 28, and 60 days after modeling, the cell apoptotic rate was significantly lower in group B than in group C (P lt; 0.05), but was significantly higher than in group A (P lt; 0.05). Conclusion 17β-estradiol can reduce the numbers of apoptotic cells and promote the nerve function recovery after chronic spinal cord injury of rats.
ObjectiveTo assess whether expanding the landing zone of frozen elephant trunk (FET) increases the risk of spinal cord injury in patients with acute type A aortic dissection. MethodsPatients with acute type A aortic dissection who were treated in Guangdong Provincial People’s Hospital from 2017 to 2020 were collected. They were divided into two groups according to the landing zone of FET by the image diagnosis of postoperative chest X-ray or total aorta CT angiography, including a Th9 group which defined as below the eighth thoracic vertebral level, and a Th8 group which was defined as above or equal to the eighth thoracic vertebral level. Using the propensity score matching (PSM) method, the preoperative and intraoperative data of two groups were matched with a 1∶2 ratio. The prognosis of the two groups after PSM was analyzed. Results Before PSM, 573 patients were collected, including 58 patients in the Th9 group and 515 patients in the Th8 group. After PSM, 174 patients were collected, including 58 patients in the Th9 group (46 males and 12 females, with an average age of 47.91±9.92 years), and 116 patients in the Th8 group (93 males and 23 females, with an average age of 48.01±9.53 years). There were 8 patients of postoperative spinal cord injury in the two groups after PSM, including 5 (4.31%) patients in the Th8 group and 3 (5.17%) patients in the Th9 group (P=0.738). In the Th8 group, 2 patients had postoperative transient paresis and recovered spontaneously after symptomatic treatment, and 1 patient had postoperative paraplegia with cerebrospinal fluid drainage. After 3 days, the muscle strength of both lower limbs gradually recovered after treatment. There was no statistical difference in complications between the two groups (P＞0.05). ConclusionExpanding the landing zone of FET does not increase the risk of spinal cord injury in patients with acute type A aortic dissection. However, the sample size is limited, and in the future, multicenter large-scale sample size studies are still needed for verification
ObjectiveTo explore the clinical characteristics and surgical treatment strategies of delayed spinal cord injury (SCI) caused by atypical compression of old thoracolumbar fracture.MethodsBetween January 2011 and June 2018, 32 patients with delayed SCI caused by atypical compression of old thoracolumbar fracture who met the inclusion criteria were admitted and divided into group A (20 cases, underwent anterior subtotal vertebral body resection+titanium mesh reconstruction+screw rod internal fixation) and group B (12 cases, underwent posterior 270° ring decompression of vertebral canal+titanium mesh reconstruction+screw rod internal fixation) according to the different operation approaches. There was no significant difference between the two groups in age, gender, cause of injury, fracture segment, disease duration, preoperative American Spinal Injury Association (ASIA) classification, and preoperative back pain visual analogue scale (VAS) score, lumbar Japanese Orthopaedic Association (JOA) score, kyphosis angle, and vertebral canal occupational ratio (P>0.05). The incision length, operation time, intraoperative blood loss, complications, and bone fusion time of reconstructed vertebrae were recorded and compared between the two groups; the kyphosis angle, back pain VAS score, and lumbar JOA score were used to evaluate the effectiveness.ResultsExcept that the incision length in group A was significantly shorter than that in group B (t=−4.865, P=0.000), there was no significant difference in intraoperative blood loss and operation time between the two groups (P>0.05). There was no deaths or postoperative paraplegia cases in the two groups, and no deep infection or skin infection occurred. There was 1 case of cerebrospinal fluid leakage, 1 case of inferior vena cava injury, and 1 case of chyle leakage in group A. No serious complications occurred in group B. There was no significant difference in the incidence of complications between the two groups (P=0.274). All 32 patients were followed up 12-61 months, with an average of 20.8 months. The follow-up time for groups A and B were (19.35±5.30) months and (23.25±12.20) months respectively, and the difference was not significant (t=−1.255, P=0.219). The reconstructed vertebrae in all cases obtained bony fusion postoperatively. The fusion time of groups A and B were (8.85±2.27) months and (8.50±2.50) months respectively, and the difference was not significant (t=0.406, P=0.688). The kyphosis angle, back pain VAS score, and lumbar JOA score of the two groups at each time point after operation and last follow-up were significantly improved when compared with preoperatively (P<0.05); the lumbar JOA score was further improved with time postoperatively (P<0.05), while the kyphosis angle and the VAS score of back pain remained similarly (P>0.05). Comparison of kyphosis angle, back pain VAS score, and lumbar JOA score between the two groups at various time points postoperatively showed no significant difference (P>0.05). At last follow-up, the JOA score improvement rate in groups A and B were 83.87%±0.20% and 84.50%±0.14%, respectively, and the difference was not significant (t=–0.109, P=0.914); the surgical treatment effects of the two groups were judged to be significant.ConclusionIn the later stage of treatment of old thoracolumbar fractures, even mild kyphosis and spinal canal occupying may induce delayed SCI. Surgical correction and decompression can significantly promote the recovery of damaged spinal cord function. Compared with anterior approach surgery, posterior approach surgery has the advantages of less trauma, convenient operation, and fewer complications, so it can be preferred.
ObjectiveTo study the effects of astaxanthin on the apoptosis after spinal cord injury in rats.MethodsOne hundred and forty-four healthy adult Sprague Dawley rats were divided into experimental group, control group, and sham group according to the random number table (n=48). In the control group and the experimental group, the modified Allen’s method was used to make the spinal cord injury model; in the sham group, only the lamina was cut without damaging the spinal cord. At immediate after operation, the rats in the experimental group were given intragastric administration of astaxanthin (75 mg/kg) twice a day; and the rats in the control group and the sham group were given equal amount of olive oil by gavage twice a day. BBB score was used to assess the motor function at 1 day and 1, 2, 3, and 4 weeks after operation. The malondialdehyde (MDA) content was determined by the thiobarbituric acid method at 24 hours after operation; and the activity of superoxide dismutase (SOD) was determined by the xanthine oxidase method. Apoptosis index (AI) was determined by TUNEL method at 6, 24, and 48 hours after operation. At 48 hours after operation, the water content of spinal cord was measured by dry-wet weight method, the lesion ratio of spinal cord was calculated, the ultrastructure of the spinal cord was observed by transmission electron microscopy, and ultrastructure scoring was performed using the Kaptanoglu score method.ResultsThe BBB score in the control group and the experimental group was significantly lower than that in the sham group at each postoperative time point (P<0.05); and the BBB score in the experimental group were significantly higher than that in the control group at 1-4 weeks postoperatively (P<0.05). The MDA content in the control group and the experimental group was significantly higher than that in the sham group at 24 hours after operation, and in the experimental group was significantly lower than in the control group (P<0.05). The SOD activity in the control group and the experimental group was significantly lower than that in the sham group, and in the experimental group was significantly higher than in the control group (P<0.05). At each time point postoperatively, the AI in the control group and the experimental group was significantly higher than that in the sham group, and in the experimental group was significantly lower than in the control group (P<0.05). At 48 hours after operation, the water content of spinal cord, the lesion ratio of spinal cord, and the ultrastructure score in the control group and the experimental group were significantly higher than those in the sham group, and in the experimental group were significantly lower than in the control group (P<0.05).ConclusionAstaxanthin can inhibit the lipid peroxidation, reduce the apoptosis, reduce the spinal cord edema, reduce the spinal cord lesion, reduce the histopathological damage after spinal cord injury, and improve the motor function of rats with spinal cord injury, and protect the spinal cord tissue, showing an obvious neuroprotective effect.
In order to investigate the clinical significance of electron-neurogram for evaluating the degree and prognosis of acute traumatic cervical spinal cord injury without fracture or dislocation, electron-neurogram and sensory evoked potential (SEP) of the upper limbs in 4 such cases were recorded from the 3rd to 30th day after the injury. The results showed SEP and MEP could be obtained from every nerve in both upper limbs, and continous monitoring of SEP and MEP could provide valuable data to judge the degree and prognosis of the injury in spinal cord.
This article investigates the role of AMP-activated protein kinase (AMPK) and its downstream signaling targets in mediating cellular processes such as autophagy, apoptosis, and inflammation, offering insights into how acupuncture may treat common central nervous system (CNS) diseases, including ischemic stroke, spinal cord injury, Parkinson disease, and Alzheimer disease. AMPK and its downstream effectors are pivotal in the signaling pathways that underlie the pathophysiology of CNS diseases. These pathways are implicated in a variety of cellular responses that contribute to the progression of neurological disorders. During CNS injury, AMPK can be activated through phosphorylation, triggering the regulation of downstream molecules and exerting protective effects on neuronal function. Acupuncture has been shown to promote neuroprotection and enhance recovery in CNS diseases through multiple mechanisms, one of which involves the activation of AMPK-related signaling pathways. Nevertheless, numerous unresolved challenges remain in this research field.