OBJECTIVE: To investigate whether Buyang Huanwu Decoction (BYHWD) had a good curative effect on the neuroprotection of red nucleus neurons after spinal cord injury (SCI) and the possible molecular mechanism. METHODS: Ninety male Sprague-Dawley rats were divided into 5 groups (n=18 per group) according to a random number table, including the control, model, low- (12.78 g/kg, BL group), medium- (25.65 g/kg, BM group), and high-dose BYHWD groups (51.30 g/kg, BH group). A rubrospinal tract transection model in rats was established, and different doses of BYHWD were intragastrically administrated for 4 weeks. The forelimb locomotor function was recorded using the spontaneous vertical exploration test. Cyclic adenosine monophosphate (cAMP) level in red nucleus was detected through an enzyme-linked immunosorbent assay. The morphology and number of red nucleus neurons were observed using Nissl's staining and axonal retrograde tracing by Fluoro-Gold (FG). The expression of cAMP-dependent protein kinase A (PKA), nuclear factor kappa-B (NF-κB) p65, and brain-derived neurotrophic factor (BDNF) in red nucleus were detected using immunohistochemistry and quantitative real-time polymerase chain reaction. RESULTS: Compared with the control group, the utilization rate of bilateral forelimbs, unilateral right forelimbs, proportion of FG-labeled positive neurons, cAMP level, protein expressions of PKA and BDNF, and BDNF mRNA expression were significantly decreased in the model group (P<0.01), while NF-κB p65 was increased in the model group (P<0.01). Compared with the model group, the utilization rate of bilateral forelimbs and unilateral right forelimbs were significantly higher in the BL, BM and BH groups (P<0.01), the proportion of FG-labeled positive neurons, cAMP level, protein expressions of PKA and BDNF and BDNF mRNA expression in all BYHWD groups were increased (P<0.05 or P<0.01), while NF-κB p65 were decreased in all BYHWD groups (P<0.05 or P<0.01). CONCLUSIONS BYHWD possesses a sound neuroprotective effect on red nucleus neurons after SCI, and the efficacy was dose-related. The mechanism may be related to regulating the cAMP/PKA/NF-κ B p65 signaling pathway, finally promoting expression of BDNF.
Animals ; Spinal Cord Injuries/pathology* ; Drugs, Chinese Herbal/therapeutic use* ; Rats, Sprague-Dawley ; Male ; Cyclic AMP/metabolism* ; Transcription Factor RelA/metabolism* ; Cyclic AMP-Dependent Protein Kinases/metabolism* ; Signal Transduction/drug effects* ; Brain-Derived Neurotrophic Factor/genetics* ; Red Nucleus/metabolism* ; Recovery of Function/drug effects* ; Neurons/metabolism* ; Rats

OBJECTIVES: Neuropathic pain (NP) is one of the most common forms of chronic pain, yet current treatment options are limited in effectiveness. Peripheral nerve injury activates spinal microglia, altering their inflammatory response and phagocytic functions, which contributes to the progression of NP. Most current research on NP focuses on microglial inflammation, with relatively little attention to their phagocytic function. Early growth response factor 2 (EGR2) has been shown to regulate microglial phagocytosis, but its specific role in NP remains unclear. This study aims to investigate how EGR2 modulates microglial phagocytosis and its involvement in NP, with the goal of identifying potential therapeutic targets. METHODS: Adult male Sprague-Dawley (SD) rats were used to establish a chronic constriction injury (CCI) model of the sciatic nerve. Pain behaviors were assessed on days 1, 3, 7, 10, and 14 post-surgery to confirm successful model induction. The temporal and spatial expression of EGR2 in the spinal cord was examined using real-time quantitative PCR (RT-qPCR), Western blotting, and immunofluorescence staining. Adeno-associated virus (AAV) was used to overexpress EGR2 in the spinal cord, and behavioral assessments were performed to evaluate the effects of EGR2 modulation of NP. CCI and lipopolysaccharide (LPS) models were established in animals and microglial cell lines, respectively, and changes in phagocytic activity were measured using RT-qPCR and fluorescent latex bead uptake assays. After confirming the involvement of microglial phagocytosis in NP, AAV was used to overexpress EGR2 in both in vivo and in vitro models, and phagocytic activity was further evaluated. Finally, eukaryotic transcriptome sequencing was conducted to screen differentially expressed mRNAs, followed by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses to identify potential downstream effectors of EGR2. RESULTS: The CCI model successfully induced NP. Following CCI, EGR2 expression in the spinal cord was upregulated in parallel with NP development. Overexpression of EGR2 via spinal AAV injection enhanced microglial phagocytic activity and increased pain hypersensitivity in rats. Both animal and cellular models showed that CCI or LPS stimulation enhanced microglial phagocytosis, which was further amplified by EGR2 overexpression. Transcriptomic analysis of spinal cord tissues from CCI rats overexpressing EGR2 revealed upregulation of numerous genes associated with microglial phagocytosis and pain regulation. Among them, Lag3 emerged as a potential downstream target of EGR2. CONCLUSIONS EGR2 contributes to the maintenance of NP by enhancing microglial phagocytosis in the spinal dorsal horn.
Animals ; Microglia/metabolism* ; Phagocytosis/physiology* ; Rats, Sprague-Dawley ; Neuralgia/physiopathology* ; Early Growth Response Protein 2/metabolism* ; Male ; Rats ; Spinal Cord/metabolism* ; Sciatic Nerve/injuries*

OBJECTIVES: To investigate the effect of monotropein on motor function recovery of mice with spinal cord injury (SCI) and explore the underlying mechanism. METHODS: Forty-five adult female C57BL/6 mice were randomized equally into sham operation group, SCI group, and SCI group with daily intraperitoneal monotropein injection. The mice in the former two groups received daily saline injections. Motor function of the mice was evaluated using BMS scores, slant plate test, and footprint analyses. Pathological changes and neuronal counts in the spinal cord were observed using HE, LFB, and Nissl staining. The biological functions of monotropein were explored using GO and KEGG enrichment analyses. NeuN/cleaved caspase-3 immunofluorescence assay and Western blotting were used to detect neuronal apoptosis in the spinal cord of the mice. In cultured HT22 cells, the effect of monotropein on TNF-α-induced cell apoptosis was evaluated using TUNEL staining and Western blotting. In monotropein-treated HT22 cells and SCI mice, the changes in the PI3K/AKT pathway were examined, and the effect of a PI3K/AKT pathway activator (IGF-1) on HT22 cell apoptosis and motor function recovery of SCI mice were observed. RESULTS: SCI mice with monotropein treatment showed significantly improved motor functions with reduced SCI areas and increased myelin retention and neuron counts in the spinal cord. Bioinformatics analysis suggested a role of PI3K/AKT signaling pathway in mediating the anti-apoptotic effects of monotropein. In SCI mice, monotropein obviously reduced apoptotic neurons, decreased expressions of cleaved caspase-3 and Bax and increased Bcl-2 expression in the spinal cord. In HT22 cells, monotropein significantly inhibited TNF-α-induced apoptosis and PI3K/AKT pathway activation. Treatment with IGF-1 obviously increased apoptosis of HT22 cells and exacerbated locomotor dysfunction in SCI mice. CONCLUSIONS Monotropein promotes motor function recovery in SCI mice by reducing neuronal apoptosis possibly by inhibiting the PI3K/AKT signaling pathway.
Animals ; Spinal Cord Injuries/metabolism* ; Apoptosis/drug effects* ; Signal Transduction/drug effects* ; Mice, Inbred C57BL ; Mice ; Proto-Oncogene Proteins c-akt/metabolism* ; Female ; Phosphatidylinositol 3-Kinases/metabolism* ; Neurons/pathology* ; Recovery of Function

OBJECTIVES: To investigate the effect of orexin-A-mediated regulation of ionotropic glutamate receptors for promoting motor function recovery in rats with spinal cord injury (SCI). METHODS: Thirty-six newborn SD rats (aged 7-14 days) were randomized into 6 groups (n=6), including a normal control group, a sham-operated group, and 4 SCI groups with daily intrathecal injection of saline, DNQX, orexin-A, or orexin-A+DNQX for 3 consecutive days after PCI. Motor function of the rats were evaluated using blood-brain barrier (BBB) score and inclined plane test 1 day before and at 1, 3, and 7 days after SCI. For patch-clamp experiment, spinal cord slices from newborn rats in the control, sham-operated, SCI, and SCI+orexin groups were prepared, and ventral horn neurons were acutely isolated to determine the reversal potential and dynamic indicators of glutamate receptor-mediated currents under glutamate perfusion. RESULTS: At 3 and 7 days after SCI, the orexin-A-treated rats showed significantly higher BBB scores and grip tilt angles than those with other interventions. Compared with those treated with DNQX alone, the rats receiving the combined treatment with orexin and DNQX had significantly higher BBB scores and grip tilt angles on day 7 after PCI. In the patch-clamp experiment, the ventral horn neurons from SCI rat models exhibited obviously higher reversal potential and greater rise slope of glutamate current with shorter decay time than those from sham-operated and orexin-treated rats. CONCLUSIONS Orexin-A promotes motor function recovery in rats after SCI possibly by improving the function of the ionotropic glutamate receptors.
Animals ; Spinal Cord Injuries/drug therapy* ; Rats ; Rats, Sprague-Dawley ; Receptors, Ionotropic Glutamate/metabolism* ; Recovery of Function/drug effects* ; Orexins/pharmacology* ; Male ; Female ; Animals, Newborn ; Neuropeptides/pharmacology* ; Intracellular Signaling Peptides and Proteins/pharmacology*

Demyelination and remyelination play key roles in spinal cord injury (SCI), affecting the recovery of motor and sensory functions. Research in rodent models is extensive, but the study of these processes in non-human primates is limited. Therefore, our goal was to thoroughly study the histological features of demyelination and remyelination after contusion injury of the cervical spinal cord in Macaca fascicularis. In a previous study, we created an SCI model in M. fascicularis by controlling the contusion displacement. We used Eriochrome Cyanine staining, immunohistochemical analysis, and toluidine blue staining to evaluate demyelination and remyelination. The results showed demyelination ipsilateral to the injury epicenter both rostrally and caudally, the former mainly impacting sensory pathways, while the latter primarily affected motor pathways. Toluidine blue staining showed myelin loss and axonal distension at the injury site. Schwann cell-derived myelin sheaths were only found at the center, while thinner myelin sheaths from oligodendrocytes were seen at the center and surrounding areas. Our study showed that long-lasting demyelination occurs in the spinal cord of M. fascicularis after SCI, with oligodendrocytes and Schwann cells playing a significant role in myelin sheath formation at the injury site.
Animals ; Spinal Cord Injuries/physiopathology* ; Demyelinating Diseases/etiology* ; Remyelination/physiology* ; Macaca fascicularis ; Disease Models, Animal ; Myelin Sheath/pathology* ; Oligodendroglia/pathology* ; Schwann Cells/pathology* ; Female ; Spinal Cord/pathology* ; Axons/pathology*

Effective axon regeneration is essential for the successful restoration of nerve functions in patients suffering from axon injury-associated neurological diseases. Certain self-regeneration occurs in injured peripheral axonal branches of dorsal root ganglion (DRG) neurons but does not occur in their central axonal branches. By performing rat sciatic nerve or dorsal root axotomy, we determined the expression of the dysbindin domain containing 2 (DBNDD2) in the DRGs after the regenerative peripheral axon injury or the non-regenerative central axon injury, respectively, and found that DBNDD2 is down-regulated in the DRGs after peripheral axon injury but up-regulated after central axon injury. Furthermore, we found that DBNDD2 expression differs in neonatal and adult rat DRGs and is gradually increased during development. Functional analysis through DBNDD2 knockdown revealed that silencing DBNDD2 promotes the outgrowth of neurites in both neonatal and adult rat DRG neurons and stimulates robust axon regeneration in adult rats after sciatic nerve crush injury. Bioinformatic analysis data showed that transcription factor estrogen receptor 1 (ESR1) interacts with DBNDD2, exhibits a similar expression trend as DBNDD2 after axon injury, and may targets DBDNN2. These studies indicate that reduced level of DBNDD2 after peripheral axon injury and low abundance of DBNDD2 in neonates contribute to axon regeneration and thus suggest the manipulation of DBNDD2 expression as a promising therapeutic approach for improving recovery after axon damage.
Animals ; Ganglia, Spinal/metabolism* ; Nerve Regeneration/genetics* ; Rats ; Axons/metabolism* ; Sciatic Nerve/injuries* ; Rats, Sprague-Dawley ; Male

OBJECTIVE: To evaluate the effectiveness of the single-stage anterior eccentric kyphotic distraction reduction technique (EKD-RT) for treating lower cervical dislocation with locked facet joints, assessing its reduction success rate, neurological improvement, and safety. METHODS: A retrospective analysis was conducted on 67 patients with lower cervical dislocation and locked facet joints (21 unilateral, 46 bilateral) treated between January 2015 and January 2024. There were 39 males and 28 females, with an average age of 49.5 years (range, 22-75 years). The injured segments included C _{3, 4} in 4 cases, C _{4, 5} in 13 cases, C _{5, 6} in 22 cases, and C _{6, 7} in 28 cases. The interval between injury and admission ranged from 2 hours to 2 days (mean, 5.6 hours). Preoperative Frankel grading included grade A in 9 cases, grade B in 28 cases, grade C in 17 cases, grade D in 11 cases, and grade E in 2 cases. Japanese Orthopaedic Association (JOA) score was 7.0±1.4. All patients underwent single-stage anterior cervical discectomy and fusion. Following discectomy at the dislocated level, the EKD-RT was applied to unlock and reduce the locked facet joints, followed by internal fixation. Operation time, blood loss, reduction success rate, and complications were recorded. Interbody fusion status was evaluated using Bridwell criteria. Neurological status was assessed pre- and post-operatively using Frankel grading. Spinal cord function was scored using the 17-point JOA score, and the improvement rate was calculated. RESULTS: Successful reduction of the locked facet joints achieved in all cases. The operation time was 41-85 minutes (range, 63.3 minutes), and intraoperative blood loss was 50-360 mL (range, 125.0 mL). Complications included cerebrospinal fluid leakage in 2 cases; no severe complications such as major vascular injury or recurrent laryngeal nerve injury occurred. All patients were followed up 12-24 months (mean, 17.9 months). At last follow-up, radiological examination confirmed interbody fusion in all patients, with no implant failure or migration. The Frankel grading included grade A in 3 cases, grade B in 9 cases, grade C in 13 cases, grade D in 16 cases, and grade E in 26 cases; the JOA score reached 13.7±2.3; all of which significantly improved compared to preoperative levels ( P<0.05). The improvement rate of JOA score was 66.1%±24.7%. CONCLUSION The EKD-RT is an effective surgical approach for lower cervical dislocation with locked facet joints. It enables safe and efficient reduction of the locked facet joints via a single incision, resulting in significant neurological improvement with a low complication rate.
Humans ; Male ; Middle Aged ; Female ; Cervical Vertebrae/diagnostic imaging* ; Retrospective Studies ; Adult ; Aged ; Zygapophyseal Joint/injuries* ; Joint Dislocations/diagnostic imaging* ; Treatment Outcome ; Spinal Fusion/methods* ; Young Adult ; Kyphosis/surgery*

OBJECTIVE: To explore the effectiveness of posterior single-level osteotomy with 360° release and correction for the treatment of osteoporotic vertebral compression fractures (OVCF) complicated with moderate to severe kyphosis. METHODS: A retrospective analysis was conducted on 11 patients with OVCF complicated with moderate to severe kyphosis between January 2022 and March 2023. There were 4 males and 7 females with an average age of 57 years ranging from 47 to 69 years. The disease duration ranged from 3 to 15 months, with an average of 7 months. Fracture segments included T ₁₁ in 3 cases, T ₁₂ in 5, L ₁ in 2, and L ₂ in 1. The T value of lumbar spine bone density was -5.0 to -2.0, with an average of -3.5. The preoperative neurological function was grade E according to Frankel grading. The Pfirrmann classification of the intervertebral disc above the injured vertebra was grade Ⅲ in 8 cases and grade Ⅳ in 3 cases. All patients underwent posterior single-level osteotomy with 360° release and correction. The operation time, intraoperative blood loss, hospital stay, and postoperative complications were recorded. Thoracolumbar local kyphosis Cobb angle, the mean height of the functional spinal unit (FSU), the sagittal vertical axis (SVA), and the sagittal index (SI) were measured. The visual analogue scale (VAS) score and Oswestry disability index (ODI) were used to evaluate the improvement of pain and function before operation, at 1 month after operation, and at last follow-up. RESULTS: The operation successfully completed in all patients, and there was no obvious complication. The operation time ranged from 100 to 190 minutes, with an average of 153 minutes, and the intraoperative blood loss ranged from 200 to 800 mL, with an average of 468 mL. The length of hospital stay was 8-14 days (mean, 12 days). All patients were followed up 6-24 months, with an average of 12.4 months. At last follow-up, all the 11 patients had bony fusion in the osteotomy area, and there was no displacement or subsidence of the Cage, no complication such as internal fixation failure or pseudarthrosis formation was found. The Cobb angle of local thoracolumbar kyphosis, the mean height of FSU, SVA, and SI significantly improved immediately after operation and at last follow-up when compared with preoperative ones, and the VAS score and ODI also significantly improved at 1 month after operation and at last follow-up ( P<0.05); there was no significant difference in above indexes between the two time points after operation ( P>0.05). CONCLUSION Posterior single-level osteotomy with 360° release and correction is an effective surgical method for treating OVCF complicated with moderate to severe kyphosis, with definite early effectiveness.
Humans ; Kyphosis/etiology* ; Male ; Female ; Osteotomy/methods* ; Middle Aged ; Retrospective Studies ; Aged ; Spinal Fractures/diagnostic imaging* ; Fractures, Compression/diagnostic imaging* ; Osteoporotic Fractures/complications* ; Treatment Outcome ; Lumbar Vertebrae/injuries* ; Thoracic Vertebrae/injuries* ; Operative Time ; Fracture Fixation, Internal/methods*

OBJECTIVE: To observe the clinical outcomes of anterior approach for the revision surgery following unsuccessful bone cement augmentation in osteoporotic vertebral compression fractures. METHODS: A total of 10 patients who experienced unsuccessful bone cement augmentation underwent anterior revision surgery between January 2020 and December 2021. There were 2 males and 8 females. The age ranged from 55 to 83 years old. The reasons for revision surgery were postoperative infection in 2 cases, postoperative neurological symptoms in 3 cases, and kyphosis resulting from postoperative vertebral collapse in 5 cases. The involved segments included 2 cases of L₁, 4 cases of L₂, 3 cases of L₃, and 1 case of L₄. Among them, 2 patients underwent primary percutaneous vertebroplasty (PVP), while eight patients underwent primary percutaneous kyphoplasty (PKP). The time interval between the first and second surgeries ranged from 1 to 13 months. All patients presented with varying degrees of lumbar pain. X-ray, CT, and MRI scans were conducted prior to the revision procedure. Surgical duration, intraoperative blood loss, and any complications were documented. RESULTS: The surgical procedures were successful in all 10 patients, with 9 cases undergoing anterior reconstruction and posterior internal fixation through the interspace of the multifidus longissimus muscle. One patient underwent anterior reconstruction and anterior plate internal fixation. The duration of the anterior approach surgery ranged from 90 to 190 minutes, with a blood loss volume ranging from 130 to 480 ml. None of the patients experienced any intraoperative or postoperative complications such as nerve injury, cerebrospinal fluid leakage, major vessel damage, abdominal organ injury, incision infection, or bleeding. The follow-up period for 10 patients ranged from 3 to 20 months. Throughout the follow-up, none of the patients experienced complications such as displacement, cutting, or loosening of the artificial vertebral body. Prior to the operation, the visual analogue scale(VAS) ranged from 4 to 8 points, the final follow-up assessment ranged from 2 to 3 points. The Oswestry disability index (ODI) score a preoperative ranged from 17% to 37%, the latest follow-up evaluation ranged from 2% to 16%. Notably, significant correction in kyphosis angle was observed in 5 cases. Furthermore, there was notable improvement in VAS and ODI compared to preoperative values. CONCLUSION Anterior corpectomy and artificial vertebral body implantation not only facilitate the removal of bone cement but also enable effective debridement and direct decompression. Moreover, by reconstructing anterior and middle column support, restoring vertebral height and local curvature, in combination with pedicle screw internal fixation, surgical segment stability can be achieved. Therefore, for patients requiring revision following failed bone cement augmentation, anterior surgery represents an ideal treatment option.
Humans ; Female ; Male ; Aged ; Bone Cements ; Fractures, Compression/surgery* ; Middle Aged ; Aged, 80 and over ; Spinal Fractures/surgery* ; Osteoporotic Fractures/surgery* ; Reoperation/methods* ; Vertebroplasty

OBJECTIVE: To explore the clinical effect of personalized puncture planning before surgery using Picture Archiving and Communication System (PACS) in the treatment of osteoporotic vertebral compression fractures in the elderly. METHODS: A total of 69 elderly patients with osteoporotic vertebral compression fractures treated by percutaneous vertebroplasty from January 2020 20 to December 2021 with more than 1 year of follow-up were analyzed retrospectively. Thirty-four patients were individualized for preoperative planning with PACS software (observation group), including 8 males and 26 females, with a mean age of (73.30±7.96) years old;and 35 patients were treated with conventional treatment (control group), including 7 males and 28 females, with a mean age of (77.30±7.84) years old. The operation time, the amount of cement injection, cement leakage rate, bone watertight diffusion and refracture within 1 year between two groups were observed and compared. The Cobb's angle, low back pain visual analogue scale(VAS) and the modified Oswsetry disability indexes(ODI) before surgery and 1 day, 1 year after surgery were compared between two groups. RESULTS: Both groups successfully completed the operation without serious surgical complications, 2 refractures occurred in the control group. The operation time in the observation group was(41.9±11.9) min, which was less than that in the control group (52.7±13.6) min (P<0.05). There was no significant difference in the cement injection volume between two groups (P>0.05). Two cases of cement leakage in the observation group was less than 8 in the control group (P<0.05). The bone cement distribution index of two groups had significant difference(P<0.05). There were no significant differences between two groups in Cobb's angle of the injured vertebras and ODI before and 1 day after surgery(P>0.05), however, the comparative differences were statistically significant at 1 year after surgery(P<0.05). There was no significant difference in the VAS between two groups at each time period(P>0.05). CONCLUSION Using the PACS software to plan personalized puncture scheme can reduce the operation time, reduce the cement leakage rate, improve the diffusion of bone cement and longer maintain the postoperative form of vertebral body and the functional state of patients' lumbar back.
Humans ; Male ; Female ; Aged ; Vertebroplasty/methods* ; Fractures, Compression/diagnostic imaging* ; Spinal Fractures/diagnostic imaging* ; Osteoporotic Fractures/diagnostic imaging* ; Aged, 80 and over ; Retrospective Studies ; Radiology Information Systems