Objective To investigate the effect of mild hypothermia on neurological function in rats with spinal cord injury (SCI) based on the adenosine monophosphate activated protein kinase (AMPK)/Nod-like receptor protein 3 (NLRP3) pathway. MethodsThe rat model of SCI was established by Allen’s method. The 7-8 weeks old SPF male SD rats were randomly divided into SCI group, mild hypothermia group (SCI+mild hypothermia treatment), Compound C group (SCI+mild hypothermia treatment+20 mg/kg AMPK/NLRP3 pathway inhibitor Compound C), and normal rats with laminectomy as sham-operation group. Basso, Beattie, and Bresnahan (BBB) score was used to evaluate the motor ability of rats at 1, 3, 7, 14 days after treatment. After 14 days, the rats were sacrificed, and the spinal cord histopathological morphology was observed by HE staining, the neuronal apoptosis in spinal cord tissue was detected by TUNEL assay, and the serum levels of interleukin 2 (IL-2), IL-6, transforming growth factor β1 (TGF-β1), malondialdehyde (MDA), and superoxide dismutase (SOD) were detected by ELISA. The expression of AMPK/NLRP3 pathway protein, including B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax), and cleaved Caspase-9 were detected by Western blot. Results At 1 day after treatment, the rats in SCI group, mild hypothermia group, and Compound C group did not recover their motor ability; at 3, 7, and 14 days, the BBB score of SCI group was significantly lower than that of SCI group (P<0.05), the BBB score of mild hypothermia group was significantly higher than that of SCI group (P<0.05), and the BBB score of Compound C group was significantly lower than that of mild hypothermia group (P<0.05). Compared with the sham-operation group, the SCI group displayed obvious pathological changes in the spinal cord tissue, with disordered tissue architecture, inflammatory infiltration, and blurred interstitial boundaries. The neuronal apoptosis rate, Bax/Bcl-2 ratio, cleaved Caspase-9 expression, NLRP3 protein expression, serum IL-2, IL-6, and MDA levels were elevated, whereas serum TGF-β1, SOD levels, and spinal cord phosphorylation AMPK/AMPK protein expression significantly decreased (P<0.05). Compared with the SCI group, the above phenomena significantly improved in the mild hypothermia group (P<0.05). Conclusion Subnormal hypothermia can attenuate neurological dysfunction after spinal cord injury in rats, potentially by activating the AMPK/NLRP3 signaling pathway.
Objective To investigate the surgery tactics for ossification of ligamentum flavum (OLF) associated with dural ossification (DO) in the thoracic spine and the cl inical outcome. Methods Between June 2006 and December 2009, 98 patients with thoracic spinal stenosis secondary to OLF were treated, and DO was found in 18 cases during operation. There were 11 males and 7 females with a mean age of 58 years (range, 46-73 years). The disease duration ranged from 5 to 48 months (mean,20 months). All patients underwent surgical decompression because of recent neurological aggravation. Both DO and OLF were resected with octagonal decompression by dissecting pedicle flavum tunnel. The Japanese Orthopaedic Association (JOA) score, modified Oswestry Disabil ity Index (ODI), and the Cobb angle were used to evaluate the effectiveness. Results The initial symptoms were significantly alleviated postoperatively. All patients had transient cerebrospinal fluid (CSF) leakage postoperatively, the CSF leakage disappeared after 8-10 days of conservative treatment. All the incisions healed by first intention. There was no complication of neurologic function deterioration, meningitis, wound infection, or spinocutaneous fistula. Eighteen patients were followed up 20-60 months (mean, 49 months). No recurrence of spinal cord compression symptoms,or neurologic function deterioration was observed at last follow-up. The JOA scores and effectiveness and modified ODI scores were significantly improved after 1 month and 12 months of operation when compared with preoperative scores (P lt; 0.05). The Cobb angles of kyphosis of the involved vertebrae were (6.7 ± 1.6)° before operation and (8.0 ± 1.2)° after 12 months of operation, showing significant difference (t=4.000，P=0.001). Postoperative T2-weighted axial MRI, sagittal MRI scan, and short T1 inversion recovery MRI showed that compressed deformity of the spinal cord returned to normal. Conclusion The surgery tactics for thoracic spinal stenosis secondary to the OLF with DO is safe, and no patching dura mater tears is effective.
Objective To study the inhibitory effect of RNA interference (RNAi) on bcl-2 expression of vascular smooth muscle cells （VSMCs) in rabbit. Methods The expression vector of bcl-2 gene-targeting small interference RNA (pshRNA-bcl-2) was constructed and was transfected into VSMCs by lipofectamine, and the unloaded vector was used as control. The expression of bcl-2 mRNA was identified by RT-PCR and Western blot, respectively. The growth of the transfected VSMCs was examined by MTT. Results The pshRNA-bcl-2 may inhibit the expression of bcl-2 gene at the levels of transcription and translation. There were significant differences (P＜0.01) of the expressions of bcl-2 mRNA between the VSMCs that were transfected with pshRNA-bcl-2 and the ones in plasmid transfected group and control group, respectively. There was a significant difference (P＜0.01) in the growth of VSMCs between the plasmid transfected and the control groups. Conclusion The plasmid containing the small interference RNA of bcl-2 may have an inhibitory effect on the cell growth and endogenous expression of bcl-2 gene at the levels of transcription and translation in VSMCs.