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.
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 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.
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 therapeutic effect of basic fibroblast growth factor (bFGF) on spinal cord injury (SCI) in rats and the influence of Notch/signal transducer and activator of transcription 3 (STAT3) signaling pathway. Methods A total of 40 10-week-old male Sprague Dawley (SD) rats were selected to establish T10-segment SCI model by a free falling object. Among them, 32 successful models were randomly divided into model group and bFGF group, with 16 in each group. Another 16 SD rats were selected as sham-operation group, with only T10 processes, dura mater, and spinal cord exposed. After modeling, the rats in bFGF group were intraperitoneally injected with 100 μg/kg bFGF (once a day for 28 days), and the rats in model group and sham-operation group were injected with normal saline in the same way. The survival of rats in each group were observed after modeling. Basso-Beattie-Bresnahan (BBB) scores were performed before modeling and at immediate, 14 days, and 28 days after modeling to evaluate the functional recovery of hind limbs. Then, the spinal cord tissue at the site of injury was taken at 28 days and stained with HE, Nissl, and propidium iodide (PI) to observe the pathological changes, neuronal survival (number of Nissl bodies) and apoptosis (number of PI red stained cells) of the spinal cord tissue; immunohistochemical staining and ELISA were used to detect the levels of astrocyte activation markers [glial fibrillary acidic protein (GFAP)] and inflammatory factors [interleukin 1β (IL-1β), tumor necrosis factor α (TNF-α), interferon γ (IFN-γ)] in tissues, respectively. Western blot was used to detect the expressions of Notch/STAT3 signaling pathway related proteins [Notch, STAT3, phosphoryl-STAT3 (p-STAT3), bone morphogenetic protein 2 (BMP-2)] in tissues. Results All rats survived until the experiment was completed. At immediate after modeling, the BBB scores in model group and bFGF group significantly decreased when compared to sham-operation group (P<0.05). At 14 and 28 days after modeling, the BBB scores in model group significantly decreased when compared to sham-operation group (P<0.05); the bFGF group showed an increase compared to model group (P<0.05). Compared with before modeling, the BBB scores of model group and bFGF group decreased at immediate after modeling, and gradually increased at 14 and 28 days, the differences between different time points were significant (P<0.05). The structure of spinal cord tissue in sham-operation group was normal; in model group, there were more necrotic lesions in the spinal cord tissue and fewer Nissl bodies with normal structures; the number of necrotic lesions in the spinal cord tissue of the bFGF group significantly reduced compared to the model group, and some normally structured Nissl bodies were visible. Compared with sham-operation group, the number of Nissl bodies in spinal cord tissue significantly decreased, the number of PI red stained cells, GFAP, IL-1β, TNF-α, IFN-γ, Notch, p-STAT3 /STAT3, BMP-2 protein expression levels significantly increased in model group (P<0.05). The above indexes in bFGF group significantly improved when compared with model group (P<0.05). Conclusion bFGF can improve motor function and pathological injury repair of spinal cord tissue in SCI rats, improve neuronal survival, and inhibit neuronal apoptosis, excessive activation of astrocytes in spinal cord tissue and inflammatory response, the mechanism of which may be related to the decreased activity of Notch/STAT3 signaling pathway.
ObjectiveTo explore the effectiveness of functional reconstruction of hand grasp and pinch by tendon transfers in patients with cervical spinal cord injury.MethodsBetween July 2013 and January 2016, tendon transfer surgery were performed in 21 patients (41 hands) with cervical spinal injury that motion level was located at C6 to reconstruct hand grasp and pinch function. There were 18 males and 3 females with a mean age of 42.3 years (range, 17-65 years). Nineteen patients were with complete spinal cord injury [American Spinal Injury Association (ASIA) grading A], 1 patient was with central cord syndrome whose bilateral hands were completely paralyzed and lower limbs were normal (ASIA grading D), and 1 patient was with cervical spondylotic myelopathy (AISA grading D). The time from injury to hospitalization was 12-22 months (mean, 16.8 months). According to the International classification of surgery of the hand in tetraplegia (ICSHT), there were 6 cases of grade O3, 10 of grade O4, 3 of grade OCu5, and 2 of grade O5. The surgery was divided into two stages with an interval of 6-11 months. At the first stage, grip function was reconstructed in all patients by transfering the extensor carpi radialis longus from radialis side to palmar side through subcutaneous tunnel, and braided and sutured with the flexor pollicis longus and flexor digitorum profundus. At the second stage, the lateral pinch function of the thumb and index finger was reconstructed by braiding and suturing the radial half of the extensor carpi ulnaris (the patients graded as ICSHT O3) or pronator tere (the patients graded above ICSHT O3) with extensor pollicis longus and abductor pollicis longus. The grasp force, the thumb and index finger lateral pinch force, and the maximum fingertips distance between the thumb and index finger were measured at preoperation and at different time points after operation. The modified Lamb and Chan questionnaire, based upon the activities of daily living, was used to evaluate the hand function of all patients at 6 months after sencond stage surgery.ResultsThere was 1 patient with elbow skin lesion, 1 patient with wrist stiffness; both of them recovered after corresponding treatment. All the 21 patients were followed up 15-32 months (mean, 19.6 months) without wound infection, tendon adhesion, tendon rupture, and other complications. The grasp forces of all patients were significantly improved at 4 weeks, 3 months, 6 months, and 1 year after the first stage surgery when compared with preoperative value (P<0.05); and no significant difference was found between different time points after operation (P>0.05). The thumb and index finger lateral pinch force and the maximum fingertips distance between the thumb and index finger of all patients were also significantly improved at 4 weeks, 3 months, 6 months, and 1 year after the second stage surgery when compared with preoperative values (P<0.05); and no significant difference was found between different time points after operation (P>0.05). And there was no significant difference of above indexes between the patients graded as ICSHT O3 and above ICSHT O3 (P>0.05). The functional outcome was good in 19 cases, fair in 1 case, and poor in 1 case according to modified Lamb and Chan questionnaire at 6 months after second stage surgery.ConclusionTendon transfer can significantly improve the hand function and the quality of life of the patients with complete cervical spinal cord injury.
Spinal cord injury (SCI) is a complex pathological process. Based on the encouraging results of preclinical experiments, some stem cell therapies have been translated into clinical practice. Mesenchymal stem cells (MSCs) have become one of the most important seed cells in the treatment of SCI due to their abundant sources, strong proliferation ability and low immunogenicity. However, the survival rate of MSCs transplanted to spinal cord injury is rather low, which hinders its further clinical application. In recent years, hydrogel materials have been widely used in tissue engineering because of their good biocompatibility and biodegradability. The treatment strategy of hydrogel combined with MSCs has made some progress in SCI repair. This review discusses the significance and the existing problems of MSCs in the repair of SCI. It also describes the research progress of hydrogel combined with MSCs in repairing SCI, and prospects its application in clinical research, aiming at providing reference and new ideas for future SCI treatment.
Objective To develop a tractive spinal cord injury model in rats with a novel spinal distractor so as to supply the rel iable animal model for researching the pathological mechanism and rehabil itation treatment of tractive spinal cordinjury. Methods A novel spinal distractor was prepared based on previous study. Sixty adult Sprague Dawley rats (weighing 250-300 g) were randomly divided into 5 groups, 12 rats in each group. T12-L3 spinal structures in the rear area were exposed and then T13-L2 spinal cords were revealed via dual laminectomy and kept integrity. In group A, a novel spinal distractor was placed without distraction; in groups B, C, D, and E, the T12-L3 spines were tracted with a novel spinal distractor which put on transverses process of T12-L3 vertebrae. During the tractive period, the somatosensory evoked potential (SEP) was used to monitor spinal cord function. The SEP ampl itudes descended 50% and kept distracting for 5 minutes in group B and for 10 minutes in group C, and descended 70% and kept distracting for 5 minutes in group D and for 10 minutes in group E, respectively to establ ish the tractive spinal cord injury model of T11-L2. The improved combine behavioral score (ICBS) was recorded at 1 and 7 days after injury in 6 rats of each group. The T13-L2 spinal tissue specimens were harvested for the morphological observation by HE and Nissl’s staining and for neurons counting. Results In group A, the ICBS score was 0 at 1 and 7 days after operation, showing significant difference when compared with the scores of the other groups (P lt; 0.05). The ICBS scores of groups D and E were significantly higher than those of groups B and C (P lt; 0.05). Edema and hemorrhage were observed in spinal cord surface and normal morphological structures were destroyed at different extent in groups B, C, D, and E at 1 day. There were adherence and congestion between spinal cord surface and peripheral issue without luster at 7 days, and dura depression was observed at the injury section, especially in group E. Necrosis and dissolution occurred in some neurons, and Nissl body structure dissolved or disappeared in groups B, C, D, and E. The neuron counting gradually decreased in accordance with the aggravation of injury in groups B, C, D, and E, showing significant difference when compared with group A (P lt; 0.05). Significant differences in neuron counting were found among groups B, C, D, and E (P lt; 0.05). Conclusion The tractive spinal cord injury model in rats can be successfully establ ished with novel spinal distractor, and the model establ ished by SEP ampl itude descending 70% and keeping distracting for 10 minutes is more suitable for study in tractive spinal cord injury.
Objective To investigate the effect of quantitative semi-transected blade on the improvement of spinal cord semi-transected and lump defect model. Methods Forty-eight male Sprague Dawley rats (weighing 220-250 g) were divided into the experimental group (n=24) and control group (n=24). The spinal cord semi-transected and lump defect model was made by self-made quantitative semi-transected blade in the experimental group, and by ophthalmic scalpel in the controlgroup. Then, the complications were observed; the electrophysiological results were detected before modeling and at 21 days after modeling; the histological changes at margin of lump defect were observed at 6 hours, 5 days, and 28 days; Basso, Beattie, and Bresnahan (BBB) scores were detected at 1, 3, 5, 7, 14, 21, 28, 35, 42, 56, and 84 days after modeling. Results There was significant difference in the mortality between the experimental group (0) and the control group (26.67%) (P=0.028). Electrophysiological examination: there was no significant difference in latency and ampl itude of motor evoked potentials (MEP) and sensory evoked potentials (SEP) between 2 groups at preoperation (P gt; 0.05); at 21 days after operation, latencies of MEP and SEP increased and the amplitude decreased in the control group, showing significant differences when compared with those in the experimental group and the preoperative values (P lt; 0.05), but no significant difference was seen between preoperation and postoperation in the experimental group (P gt; 0.05). Histological examination: in the control group, small hematoma could be observed at normal side at 6 hours after modeling, increased spaces of spinal tissue and perineural invasion were observed at 5 days, and small cavity formed without normal motoneurons at 28 days in the margin of lump defect. In the experimental group, no small hematoma could be observed at 6 hours after modeling, no inreversible injury of neuron and small cavity were observed at 5 days, and normal motoneurons were observed without small cavity at 28 days in the margin of lump defect.BBB scores: except the scores between experimental group and control group at affected side (P gt; 0.05), there were significant differences between groups, and between normal side and affected side for intragroup (P lt; 0.05). Conclusion Semi-transected and lump defect model could be set up successfully by self-made quantitate semi-transected blade, procedure is repetitive and the model is stable. This model is an ideal model for semi-transected spinal cord injury.
ObjectiveTo systematically profile and characterize the circular RNA (circRNA) and microRNA (miRNA) expression pattern in the lesion epicenter of spinal tissues after traumatic spinal cord injury (TSCI) and predict the structure and potential functions of the regulatory network.MethodsForty-eight adult male C57BL/6 mice (weighing, 18-22 g) were randomly divided into the TSCI (n=24) and sham (n=24) groups. Mice in the TSCI group underwent T8-10 vertebral laminectomy and Allen’s weight-drop spinal cord injury. Mice in the sham group underwent the same laminectomy without TSCI. The spinal tissues were harvested after 3 days. Some tissues were stained with HE staining to observe the structure. The others were used for sequencing. The RNA-Seq, gene ontology (GO) analysis, and circRNA-miRNA network analyses (TargetScan and miRanda) were used to profile the expression and regulation patterns of network of mice models after TSCI.ResultsHE staining showed the severe damage to the spinal cord in TSCI group compared with sham group. A total of 17 440 circRNAs and 1 228 miRNAs were identified. The host gene of significant differentially expressed circRNA enriched in the cytoplasm, associated with positive regulation of transcription and protein phosphorylation. mmu-miR-21-5p was the most significant differentially expressed miRNA after TSCI, and circRNA6730 was predicted to be its targeted circRNA. Then a potential regulatory circRNA-miRNA network was constructed.ConclusionThe significant differentially expressed circRNAs and miRNAs may play important roles after TSCI. A targeted interaction network with mmu-miR-21-5p at the core of circRNA6730 could provide basis of pathophysiological mechanism, as well as help guide therapeutic strategies for TSCI.