Basso, Beattie And Bresnahan Scores Research Articles

Effect of neuroglobin gene-modified bone marrow mesenchymal stem cells on spinal cord injury in rats

Objective To observe the protective effect of neuroglobin gene-modified bone marrow mesenchymal stem cells(BMSCs)on spinal cord injury(SCI)in rats and explore the mechanism. Methods Sixty SD rats were randomly divided into saline group, BMSCs group, Neuroglobin(Ngb)/BMSCs group and blank group, given saline, BMSCs-culture medium, BMSCs-culture medium infected with Ngb after SCI,and nothing respectively.Measurements were evaluated by the Basso,Beattie and Bresnahan (BBB)scores at 1st,3rd,7th,14th and 21st day separately Results The BBB scores at 1st and 3rd day in each group were as follows:blank group(2. 76±0. 50,and 3. 70±0. 76),saline group(2. 72±0. 53,and 4. 06±0. 77), BMSCs group(2. 76±0. 71,and 4. 13±0. 76),and Ngb/BMSCs group(2. 79±4. 20,and 4. 20±0. 82). There was no significant difference among groups. At 7th, 14th and 21st day, the BBB scores in each group were gradually increased:blank group(5. 22±0. 64,6. 16±0. 78,and 7. 54±0. 90), saline group(5. 72±0. 94, 7. 61±0. 76, and 10. 87±0. 91), BMSCs group(5. 87±0. 84, 8. 33±0. 94, and 11. 45±1. 02), and Ngb/BMSCs group(6. 52±0. 95, 9. 78±0. 84, and 13. 33±1. 05).There were statistically significant difference between Ngb/BMSCs group and others(P< 0. 05). Conclusion Neuroglobin gene-modified BMSCs can make injured spinal cord supply stable and a high level Ngb in time, which can obviously help recovery of neural function after SCI in rats. Key words: Neuroglobin; Gene modified; Bone marrow mesenchymal stem cells; Spinal cord injury

Comparative Study on the Effects of Ceftriaxone and Monocytes on Recovery after Spinal Cord Injury in Rat.

Comparison between the efficacy of ceftriaxone and monocytes on improvement of neuron protection and functional recovery after spinal cord injury (SCI) in rat. Rats were randomly divided into three groups of ten. Spinal cord injury was performed on rats under general anesthesia using the weight dropping method. Ceftriaxone was injected intraperitoneally 200 mg/kg/day for seven days after SCI. Monocytes were injected 2 × 105 cells 4 days after SCI. Hind limb motor function was assessed using the Basso, Beattie and Bresnahan (BBB) scale. Corticospinal tract (CST) axons were traced by injection of biotin dextran amine (BDA) into the sensorimotor cortex. There were statistically significant differences in BBB scores in ceftriaxone in comparison to both monocytes receiving and control groups. On the other hand there were statistically significant differences in axon counting in both ceftriaxone and monocytes receiving groups in comparison to control group. Our findings suggest that ceftriaxone improves functional recovery more effective than monocytes in rats after SCI. These results are from an experimental model and validation is required for further investigation.

An Acute Growth Factor Treatment that Preserves Function after Spinal Cord Contusion Injury

Inflammation of the spinal cord after traumatic spinal cord injury (SCI) leads to destruction of healthy tissue. This "secondary degeneration" is more damaging than the initial physical damage and is the major contributor to permanent loss of functions. In our previous study, we showed that combined delivery of two growth factors, vascular endothelial growth factor and platelet-derived growth factor, significantly reduced secondary degeneration after hemisection injury of the spinal cord in the rat. Growth factor treatment reduced the size of the lesion cavity at 30 days, compared to control animals, and further reduced the cavity at 90 days in treated animals, whereas in control animals the lesion cavity continued to increase in size. Growth factor treatment also reduced astrogliosis and reduced macroglia/macrophage activation around the injury site. Treatment with individual growth factors alone had similar effects to control treatments. The present study investigated whether growth factor treatment would improve locomotor behavior after spinal contusion injury, a more relevant pre-clinical model of SCI. The growth factors were delivered for the first 7 days to the injury site by osmotic minipump. Locomotor behavior was monitored at 1-28 days after injury using the Basso, Beattie and Bresnahan (BBB) score and at 30 days using automated gait analysis. Treated animals had BBB scores of 18; control animals scored 10. Treated animals had significantly reduced lesion cavities and reduced macroglia/macrophage activation around the injury site. We conclude that growth factor treatment preserved spinal cord tissues after contusion injury, thereby allowing functional recovery. This treatment has the potential to significantly reduce the severity of human spinal cord injuries.

Repair of Spinal Cord Injury (SCI) Using Bone Marrow Stromal Cell Transfected with Adenoviral Vector Expressing Glial derived Neurotropic Factor (GDNF) in a Rat SCI Model

Back ground Subsequent to spinal cord injury many pathological changes may occur that could lead to inappropriate environment for repair. The Most important of such changes is the death of neurons. Exogenous administration of growth factors that modulate neuronal survival, synaptic plasticity, and neurotransmission has been proposed as a potential therapeutic treatment for SCI. Among these growth factors, GDNF is a peptide with pleiotropic survival and growth-promoting effects on neurons. In addition, GDNF induces the growth of motor and sensory axons and inhibits neuronal apoptosis. Adult stem cells may provide new strategies to treat SCI. Among various types of candidate stem cells, bone marrow stromal cells (BMSC) are promising because they have shown potential to neuronal differentiation and repair in damaged spinal cord. In this study, we aimed to improve results of treatment using combination of BMSC and GDNF features. Methods: Rats were divided randomly into four groups of six. Spinal cord injury was then performed under general anesthesia using the weight dropping method. The BMSCs were injected on 3th day of post-spinal cord injury. Group one included rats receiving normal saline, group two received BMSC, group three received BMSC infected with adenoviruses encoding the beta-galactosidase gene, and group four received BMSC infected with adenoviruses encoding the GDNF gene. A Basso, Beattie and Bresnahan (BBB) score test was performed for a period of four weeks. Two weeks before the end of BBB, biotin dextran amine was injected intracerebrally followed by tissue staining at the end of the fourth week. Results: There was a significant difference in BBB scores between groups one and four (p<0.05). BBB scale improved to 12.8 points group one, while it was 8.6 in the control. There were no significant differences between other groups. There were significant differences in axon counting between group one and other groups (p<0.0001). Also, there were significant differences in axon counting between group four with groups two and three (p<0.0001). Conclusion: Combined use of these methods (GDNF expressing BMSCs) showed better results in comparison with BMSC alone. In this study the two methods were used simultaneously. The time of injection of BMSCs is very important and so we suggest that other injection times be tested. Key words: BMSC, GDNF, Gene Therapy, Spinal cord injury.

Effect of asiatic acid on oxidative stress after acute spinal cord injury in rats

Objective To observe the effect of asiatic aicd treatment on acute spinal cord injury in rats.Methods Forty-five SD rats were randomly divided into a sham-operation group,model control group and asiatic acid treatment group.Functional outcome measurements were evaluated by the Basso,Beattie and Bresnahan (BBB) score at 24,48 and 72 h.The sections were stained with hematoxylin and eosin for morphological and histopathological analyses.The reactive oxidative species and inflammatory factor were detected.Results The BBB score was significantly higher in the asiatic acid treatment group (6.70 ± 1.09 and 9.58 ±2.81) than in the model control group (3.05 ±0.98 and 4.55 ±0.65) at 48 h and 72 h (P ＜ 0.05).Compared to the model control,HE staining showed the histopathological performance was improved greatly in rat spinal cord in asiatic acid treatment group.At the three time points,tissue malondialdehyde (MDA) content in asiatic acid treatment group [(6.42 ± 0.15),(6.17 ± 0.25) and (5.48 ± 0.33) nmol/ml] was statistically significantly lower than in the model group [(7.55 ±0.37),(7.61 ± 0.24) and (6.59 ±0.22) nmol/ml].The superoxide dismutase (SOD) activity in the asiatic acid treatment group [(220.55 ±3.76),(231.44 ±4.88) and (235.98 ±5.26) U/mg] was higher than in the model group [(180.31 ±4.68,175.32 ±3.45 and 170.51 ±3.44) U/mg] at the three time points.After injury,tissue myeloperoxidase (MPO) and IL contents were increased,and showed a significant difference between the model control group and asiatic acid treatment group at the three time points.Conclusion The present study suggests that asiatic acid exerts neuroprotection by reducing oxidative stress. Key words: Asiatic acid; Spinal cord injury; Oxidative stress

Riluzole improves outcome following ischemia–reperfusion injury to the spinal cord by preventing delayed paraplegia

The spinal cord is vulnerable to ischemic injury due to trauma, vascular malformations and correction of thoracic aortic lesions. Riluzole, a sodium channel blocker and anti-glutamate drug has been shown to be neuroprotective in a model of ischemic spinal cord injury, although the effects in clinically relevant ischemia/reperfusion models are unknown. Here, we examine the effect of riluzole following ischemia–reperfusion injury to the spinal cord.Female rats underwent high thoracic aortic balloon occlusion to produce an ischemia/reperfusion injury. Tolerance to ischemia was evaluated by varying the duration of occlusion. Riluzole (8mg/kg) was injected intraperitoneally 4h after injury. Locomotor function (Basso, Beattie and Bresnahan (BBB) scale) was assessed at 4h, 1day, and 5days post-ischemia. Spinal cords were extracted and evaluated for neuronal loss using immunohistology (choline acetyltransferase (ChAT) and neuronal nuclei (NeuN)), inflammation (CD11b), astrogliosis (glial fibrillary acidic protein – GFAP) and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL).Ischemic injury lasting between 5.5 and 6.75min resulted in delayed paraplegia, whereas longer ischemia induced immediate paraplegia. When riluzole was administered to rats that underwent 6min of occlusion, delayed paraplegia was prevented. The BBB score of riluzole-treated rats was 11.14±4.85 compared with 1.86±1.07 in control animals. Riluzole also reduced neuronal loss, infiltration of microglia/macrophages and astrogliosis in the ventral horn and intermediate zone of the gray matter. In addition, riluzole reduced apoptosis of neurons in the dorsal horn of the gray matter.Riluzole has a neuroprotective effect in a rat model of spinal cord injury/reperfusion when administered up to 4h post-injury, a clinically relevant therapeutic time window.

Functional electrical stimulation post-spinal cord injury improves locomotion and increases afferent input into the central nervous system in rats

BackgroundFunctional electrical stimulation (FES) has been found to be effective in restoring voluntary functions after spinal cord injury (SCI) and stroke. However, the central nervous system (CNS) changes that occur in as a result of this therapy are largely unknown.ObjectiveTo examine the effects of FES on the restoration of voluntary locomotor function of the CNS in a SCI rat model.MethodsSCI rats were instrumented with chronic FES electrodes in the hindlimb muscles and were divided into two groups: (a) FES therapy and (b) sedentary. At day 7 post-SCI, the animals were assessed for locomotion performance by using a Basso, Beattie and Bresnahan (BBB) scale. They were then anesthetized for a terminal in vivo experiment. The lumbar spinal cord and somatosensory cortex were exposed and the instrumented muscles were stimulated electrically. Associated neurovascular responses in the CNS were recorded with an intrinsic optical imaging system.ResultsFES greatly improved locomotion recovery by day 7 post-SCI, as measured by BBB scores (P < 0.05): (a) FES 10 ± 2 and (b) controls 3 ± 1. Furthermore, the FES group showed a significant increase (P < 0.05) of neurovascular activation in the spinal cord and somatosensory cortex when the muscles were stimulated between 1 and 3 motor threshold (MT).ConclusionHind limb rehabilitation with FES is an effective strategy to improve locomotion during the acute phase post-SCI. The results of this study indicate that after FES, the CNS preserves/acquires the capacity to respond to peripheral electrical stimulation.

Effects of the combined administration of vitamins C and E on the oxidative stress status and programmed cell death pathways after experimental spinal cord injury

Experimental, controlled, animal study. To assess the effects of vitamins C and E (VCE) treatment on oxidative stress and programmed cell deaths after rat spinal cord injury (SCI), as well as functional recovery. Taiwan. Fifty-four Sprague-Dawley rats were used for the experimental procedure. In the sham group, laminectomy at T10 was performed, followed by impactor contusion of the spinal cord. In the control group, only a laminectomy was performed without contusion. Oxidative stress status was assessed by measuring the spinal cord tissue content of superoxide dismutase (SOD) and gluthatione peroxidase (GSH-Px) activities. We also evaluated the effects of combined VCE treatment using western blot to analyze expression of cleaved caspase-3 and microtubule-associated protein light chain 3 (LC3), and the Basso, Beattie and Bresnahan (BBB) scale to evaluate functional outcomes. Combined treatment of VCE significantly counteracted the effects of spinal cord contusion on oxidative stress represented by activities of SOD and GSH-Px (P<0.05). The VCE treatment also significantly enhanced LC3-II expression and decreased cleaved caspase-3 compared with the sham (P<0.05). Furthermore, BBB scores significantly improved in the VCE-treated group compared with the sham group (on day 14 and 28 after SCI; P<0.05). The combined administration of VCE was clearly capable of modulating the antioxidant effects, and of reducing apoptosis and increasing autophagy at the lesion epicenter leading to an improved functional outcome. Use of such clinically ready drugs could help earlier clinical trials in selected cases of human SCIs.

Bone marrow stromal cells inhibit caspase-12 expression in rats with spinal cord injury

The mechanisms underlying the potentially beneficial effect of bone marrow stem cells (BMSCs) on spinal cord injury (SCI) are unknown. Therefore, the aim of the present study was to explore the protective effect of BMSCs in rats with SCI. A total of 45 adult male Sprague-Dawley rats were randomly divided into three groups; the SCI group (n=15), the BMSC group (n=15) and the sham-operation group (n=15). In the SCI and BMSC treatment groups, a modified Allen’s weight-drop technique was used to induce SCI. The BMSC treatment group received an injection of BMSCs using a microneedle into the epicenter of the spinal cord 24 h after injury. Rats in the sham-operation group were not subjected to SCI; however, the corresponding vertebral laminae were removed. Seven days after transplantation, a rapid recovery was observed in the Basso, Beattie and Bresnahan (BBB) scores of the BMSC treatment group, whereas the BBB scores in the SCI group remained low (P<0.05). Caspase-12 expression in the SCI group was increased compared with that in the sham-operation group, whereas caspase-12 expression was attenuated 24 h after transplantation in the BMSC treatment group (P<0.05). In conclusion, the transplantation of BMSCs may improve locomotor function and attenuate caspase-12 expression following SCI. Therefore, it is likely to be an effective strategy for preventing severe injury of the spinal cord.

An experimental spinal cord injury rat model using customized impact device: A cost-effective approach

Till date, NYU MASCIS (New York University, Multicenter Animal Spinal Cord Injury Study) impactor and Ohio State University electromagnetic spinal cord injury device impactor were under use for simulating an experimental spinal cord injury in rodents; functional recovery being assessed through Basso, Beattie and Bresnahan (BBB) scoring method which is an open field behavior based scoring system. Although, the cited impactors are state-of-art devices, affordability to scientists in developing and under developed countries is questionable. Since the acquisition of these impact devices are expensive, we designed a customized impact device based on the requirement, satisfying all the parameters to withstand a standard animal model for contusion type of spinal cord injury at the thoracic level without compromising the lesion reproducibility. Here, a spinal cord contusion is created using a blunt-force impactor in male Wistar rats. Our method gave consistent lesion effects as evaluated by behavior scoring methods. All the animals showed equal degree of performance in tests like narrow beam, inclined plane and horizontal ladder and in BBB scores (open field locomotor test). The aim of presenting our experience is to reinstate the fact that lack of affordability to get sophisticated instrumentation need not be a hurdle in the pursuit of science.

Alginic acid sodium hydrogel co-transplantation with Schwann cells for rat spinal cord repair.

IntroductionThe aim of the study was investigating the influence of Schwann cells-alginic acid sodium hydrogel co-transplantation on a rat model of spinal cord injury.Material and methodsSprague-Dawley (SD) rats were randomly assigned to 4 groups: control, injury, injury with Schwann cell transplantation, and injury with Schwann cells-alginic acid sodium hydrogel co-transplantation. Gelatin sponge blocks containing a Schwann cell suspension were transplanted into the injury site in the Schwann cell group; Schwann cells seeded in alginic acid sodium hydrogel were transplanted into the injury site in the Schwann cells-alginic acid sodium hydrogel group. At 12 h, 1, 3, 7, and 21 days after surgery, animals were assessed on the Basso, Beattie and Bresnahan (BBB) locomotor rating scale and then were sacrificed.ResultsIn the injury group, Bcl-2 immunoreactive cells peaked at 3 days after surgery, and the expression level returned to normal level at 14 days. In the co-transplantation group, Bcl-2 immunoreactive cells in the spinal cord-transected segments were significantly increased until 7 days (p < 0.05) and remained at this level for more than 14 days. In the injury group, the number of apoptotic cells was the highest, as compared with the other 3 groups, and peaked at 1 and 7 days following spinal cord injury, and they were mostly distributed in the white matter. The BBB scores were significantly higher in the Schwann cells-alginic acid sodium hydrogel transplantation group than in the simple injury and Schwann cell groups (p < 0.05).ConclusionsSchwann cells-alginic acid sodium hydrogel co-transplantation could inhibit cellular apoptosis and enhance Bcl-2 expression in the spinal cord-transected segments, and thereby promote the recovery of locomotor function after spinal cord injury, although it did not reach full rehabilitation.

Effective reinnervation of the quadriceps femoris by spinal ventral root cross-anastomosis in rats

To study the effective recovery of the quadriceps femoris by spinal ventral root cross-anastomosis in rats. End-to-end anastomosis was performed between the left L(1) and L(3) ventral roots using autogenous nerve graft, and the right L(1) and L(3) roots were left intact. In control animals, the left L(3) ventral root was cut and shortened, and anastomosis was not performed. Six months postoperatively, the movement of low extremities was detected by electrophysiological examination, hindlimb locomotion and basso, beattie and bresnahan (BBB) scoring at one, three, seven, 14, 21 and 28 days after SCI. Fluorescence retrograde tracing with TRUE BLUE (TB) and HE staining were performed to observe the nerve regeneration. Six months after surgery, the anastomotic nerve was smooth and not atrophic. The amplitudes of action potential were 7.63 ± 1.86 mV and 6.0 ± 1.92 mV respectively before and after the spinal cord hemisection. The contraction of left quadriceps femoris was induced by a single stimulation of the anastomotic nerve. The locomotion of left hindlimb was partially restored after spinal cord hemisection while creeping and climbing. In addition, there was significant difference in the BBB score at one, three and seven days after SCI. TB retrograde tracing and neurophysiologic observation indicated efficient reinnervation of the quadriceps femoris. The cross-anastomosis between spinal ventral root can partially reconstruct the function of quadriceps femoris following SCI and may have clinical implication for the treatment of human SCI.

Preventing Venous Thromboembolism After Thoracic or Thoracolumbar Spinal Fusion

The spinal cord is vulnerable to ischemic injury due to trauma, vascular malformations and correction of thoracic aortic lesions. Riluzole, a sodium channel blocker and anti-glutamate drug has been shown to be neuroprotective in a model of ischemic spinal cord injury, although the effects in clinically relevant ischemia/reperfusion models are unknown. Here, we examine the effect of riluzole following ischemia–reperfusion injury to the spinal cord.Female rats underwent high thoracic aortic balloon occlusion to produce an ischemia/reperfusion injury. Tolerance to ischemia was evaluated by varying the duration of occlusion. Riluzole (8 mg/kg) was injected intraperitoneally 4 h after injury. Locomotor function (Basso, Beattie and Bresnahan (BBB) scale) was assessed at 4 h, 1 day, and 5 days post-ischemia. Spinal cords were extracted and evaluated for neuronal loss using immunohistology (choline acetyltransferase (ChAT) and neuronal nuclei (NeuN)), inflammation (CD11b), astrogliosis (glial fibrillary acidic protein – GFAP) and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL).Ischemic injury lasting between 5.5 and 6.75 min resulted in delayed paraplegia, whereas longer ischemia induced immediate paraplegia. When riluzole was administered to rats that underwent 6 min of occlusion, delayed paraplegia was prevented. The BBB score of riluzole-treated rats was 11.14 ± 4.85 compared with 1.86 ± 1.07 in control animals. Riluzole also reduced neuronal loss, infiltration of microglia/macrophages and astrogliosis in the ventral horn and intermediate zone of the gray matter. In addition, riluzole reduced apoptosis of neurons in the dorsal horn of the gray matter.Riluzole has a neuroprotective effect in a rat model of spinal cord injury/reperfusion when administered up to 4 h post-injury, a clinically relevant therapeutic time window.

Panax notoginsenoside produces neuroprotective effects in rat model of acute spinal cord ischemia–reperfusion injury

Ethnopharmacological relevanceAcute spinal cord ischemia–reperfusion injury (SCII) is associated with pathological changes, including inflammation, edema, and neuronal apoptosis. Panax notoginsenoside (PNS), an important traditional Chinese medicine, has shown a variety of beneficial effects, including homeostasis maintenance, anti-myocardial ischemia activities, and neuroprotective functions. However, whether it can produce neuroprotective effects in SCII and the underlying mechanisms remain largely elusive. Aim of the studyIn the present study, we investigated the effects of PNS on neurological and histopathological changes after SCII as well as the underlying mechanisms. Materials and methodsSixty-four adult rats were randomly assigned into one of the four groups: the sham group, the ischemic group, the PNS group, and the Methylprednisolone group. A rat model of SCII was adopted from a commonly used protocol that was initially proposed by Zivin. Neurological function was evaluated with the Basso, Beattie and Bresnahan (BBB) locomotor rating scale. Histopathological changes were examined with hematoxylin and eosin staining as well as Nissl staining. Immunohistochemistry and Western blot were conducted to compare the changes in tumor necrosis factor-α, interleukin-1β, interleukin-10, aquaporin-4 (AQP-4), member 6 of the TNF receptor superfamily (Fas), and Fas ligand (FasL) in the spinal cord. Finally, neuronal apoptosis was measured by electron microscopy. ResultsThe BBB scores of the PNS-treated injured animals were significantly increased. The gross histopathological examination showed restored neuronal morphology and increased number of neurons after the PNS treatment. The PNS treatment decreased SCII-induced up-regulation of cytokine levels. In addition, PNS suppressed the increased expression of AQP-4 after SCII, suggesting an anti-edema effect. Finally, PNS treatment inhibited injury-induced apoptosis and reduced the expression levels of apoptosis-related proteins, Fas and FasL, confirming its anti-apoptosis effects against SCII. ConclusionThe current findings suggest that PNS produces robust neuroprotective effects in spinal cord ischemia–reperfusion injury, and this role may be mediated by its anti-inflammation, anti-edema, and anti-apoptosis actions.

Effect of erythropoietin on the mobility of bone marrow mesenchymal stem cells to the sites of injured spinal cord

Objective To obsevre the effect of erythropoietin (EPO) treatment on the mobility of bone marrow mesenchymal stem cell (BMSCs) to the sites of injured spinal cord and explore the molecular mechanism. Methods Thirty SD rats were randomly divided into sham-operation group, saline group and EPO treatment group. BMSCs labeled with Hoechst33342 were transplanted by tail vein. Functional outcome measurements were evaluated by the Basso, Beattie and Bresnahan (BBB) score at 1st, 3rd, 7th,14th, 21st and 28th day. The immigrating number of BMSCs was measured by immunofluorescent staining.The expression of brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF) in BMSCs was detected by Western blotting. Results From the 3rd day on, the BBB scores in EPO treatment group were (5.29±0.69), (8.18±0.38), (13.32±0.77), (15.25 ±1.83), (18.71 ±1.54) respectively, which were obviously higher than in saline group. The number of BMSCs was (90. 12 ±7. 68), (116. 26 ± 13. 54),( 186. 32 ± 20. 35 ), (211.64 ± 31.83 ) and (298.71 ± 21.54) respectively, which was significantly increased in EPO treatment group (P ＜ 0. 05 ). The protein levels of BDNF and NGF expressed by BMSCs were increased significantly (P ＜ 0. 05) in EPO treatment group as compared with sham-operation group.Conclusion EPO treatment after spinal cord injury may help BMSCs migrate to the injury site and accelerate recovery of neural function by up-regulating the expression of BDNF and NGF. Key words: Erythropoietin; Bone marrow mesenchymal stem cell; Spinal cord injury

The effect of different dose of butorphanol with epidural injections on neurological function for rats

Objective To observe the effect of epidural injection with different dose of butorphanol on the rats' neurological function.Methods A PE-530 catheter was inserted into the epidural space of all the SpragueDawley rats (male, weighting 180 ～210 g) at L1-2 level.After three days, a total of 32 rats without any motor dysfunction were randomly divided into 4 groups as follows saline(NS) group (group C, n= 8 )and butorphanol injection (B) group( B1∶ n=8;B2∶ n=8;B3∶ n=8).Rats in group C were epidurally injected NS 30 μl each ,and rats in group B1, B2 and B3 were respectively epidurally injected Butorphanol 60 μg/30μl, 120 μg/30 μl,240 μg/30 μl (all diluted with NS) ,and 1 time per day for5 days.The neurological function of rats was recorded before injection (T0) and 6h after injection on day 1 ～4(T1 ～T 4) and 6h,24h and 72h after injection on day 5 (T5 ～T7) by BBB (BASSO,BEATTIE and BRESNAHAN ) Score and the inclined plane test .Results Compared with group C ,the BBB score and the inclined plane test of group B1 showed no significant difference throughout the experimental period(P＞ 0.05 ).There was also no significant difference at T0 ～ T3 of group B2 and group B3 compared with group C (P ＞ 0.05 ), while at T4, the BBB score ( ( 18.50 ± 2.00 ) points, ( 16.38 ± 2.33 ) points) and the inclined plane test( (58.75 ± 5.17 )°, (59.38 ± 3.20) ° ) of the two groups were both obviously decreased when compared with group C( (21.00 ±0.00) points, (65.00 ±3.78)°, P＜0.05) ,and the same significant differences appeared at T5,T6 and T7 (P ＜ 0.05 ).Conclusion Repeated epidural injection of butorphanol 60 μg have no effect on neurological function of rats,while repeated epidural injection of butorphanol 120 μg and 240 μg could impaire the neurological function. Key words: Butorphanol; Epidural; BBB score; The inclined plane test

The effect of olfactory ensheathing cell transplantation combined with walking training on neurofunction recovery in rats after spinal cord contusion

Objective To explore the effect of olfactory ensheathing cell （OEC） transplantation combined with walking training on neurofunction recovery in rats after spinal cord contusion. Methods Forty adult female rats aged （75 ± 1 ） days were subjected to experimental spinal cord contusion at the T10 level using a New York University impactor at a height of 25 mm. They were then divided into 4 groups： （ 1 ） an OEC transplantation combined with walking training （OEC-walking training） group, （2） an OEC transplantation （OEC） group, （3） a walking training combined with Dulbecco＇s modified Eagle medium injection （DMEM） （walking training-DMEM） group, and （4） aDMEM injection （SCI-DMEM） group. The OEC transplants and DMEM injections were performed 2 weeks post-injury. Walking training began at the 7th day post-injury and consisted of daily sessions （once daily, 5 days a week for 10 weeks） of quadrupedal treadmill training, starting from 15 min and gradually increasing to 30 min daily, at speeds starting from 3 m/min and gradually increasing in accordance to the condition of the rats. Locomotor function recovery of the rats＇ hindlimbs was evaluated weekly using the Basso, Beattie and Bresnahan （BBB） locomotor rating scale.The expression of tyrosine hydroxylase （ TH ） was detected in the injured region of the lumbar spinal cord. Results The BBB scores of rats in the OEC-walking training group and the walking training-DMEM group improved significantly from the 4th week post-injury compared to the SCI-DMEM injection group. Rats in the OEC transplantation group had a significant improvement in BBB scores at the 5th to 8th weeks post-injury. At the end of the 11th week post-injury, the average BBB scores were 13.14 ± 0.24 in the OEC-walking training group, 11. 64 ± 0.56 in the OEC transplantation group, 12.29 ±0.64 in the walking training-DMEM group and 11.07 ± 0.84 in the SCI-DMEM group.The OEC-walking training group scored significantly higher than the other 3 groups. Although the number of TH-positive neurons in the lumbar spinal cord was not significantly different among the groups, the morphology of TH-positiveneurons in the OEC-walking training group and the walking training-DMEM group was different from those in the OEC transplantation group and the SCI-DMEM group. Conclusions OEC transplantation combined with walking training can effectively promote the functional recovery of the hindlimb. The plasticity of the descending TH system and of motoneurons of the ventral horn of the lumbar spinal cord might mediate the changes. Key words: Spinal cord contusion; Olfactory ensheathing cells transplantation; Walking training; Neurofunction; Tyrosine hydroxylase

ScoreCentre: A computer program to assist with collection and calculation of BBB locomotor scale data

The Basso, Beattie and Bresnahan (BBB) Locomotor Rating Scale is a standardized assessment scale for use in experimental spinal cord injury (SCI) research. This paper describes a computer program, ScoreCentre, which aims to simplify the recording and handling of BBB locomotor scale data. The program assists with the recording of observational data from open-field testing and then automatically calculates BBB scores. Possible errors associated with data entry and manual calculation of scores are thus essentially eliminated. In addition, significant time is saved by the automated derivation of scores and subscores and elimination of the need to manually transfer data from paper records to a computer. ScoreCentre can also be used as a training aid, to help familiarize users with the BBB scale and to explore how changes in the observations impact on overall BBB score. ScoreCentre includes simple experiment management functions such as control of trial blinding, administration of drugs in a blinded fashion and longitudinal data analysis. ScoreCentre provides all the advantages of electronic records, such as ease of use, analysis and archiving, and allows the elimination of paper records if appropriate. When paper records are required, for example for archiving and auditing, they can be automatically produced by ScoreCentre. ScoreCentre will assist with both the learning and use of the BBB locomotor scale, thus facilitating the use of this standardized outcome measure in SCI research. S coreCentre is available to download from www.rmeonline.net/scorecentre.

Analysis on the Result of Wnt-3a Producing Fibroblast Implantation in Spinal Cord Injury of Rat

Objective: Promoting axonal regeneration is one of essential goals to be achieved for effective repair from spinal cord injury (SCI). The authors performed an experiment on the effect of Wnt-3a producing fibroblast injection on SCI. Methods: SCI is performed by New York University (NYU) impactor in 21 rats, and 1 week after injury, the rats were grouped into Wnt-3a producing fibroblast transplantation group (Wnt group, n=7), fibroblast transplantation group (vehicle group, n=7), and no treatment group (sham group, n=7). For the behavioral tests, the Basso, Beattie and Bresnahan (BBB) locomotor rating scale were evaluated. Also, to study radiological evidence of axon regeneration, the rats underwent manganese enhanced MRI (ME-MRI) twice, before treatment and 5 weeks after treatment. Results: On average, the sham group and vehicle group rats attained BBB score of 10.4 and 12.1, respectively, whereas Wnt group rats achieved a significantly higher BBB score of 14.0 at 5 weeks after treatment. At 5 weeks after treatment, relative signal intensities of ME-MRI was significantly higher in the Wnt group than in the vehicle group and sham group at 5 weeks after injury. Conclusion: In this study, the authors observed that neurological recovery after SCI in rats were improved by Wnt-3a producing fibroblasts which were injected intramedullary at 1 week after SCI. (J Kor Neurotraumatol Soc 2010;6:1-5)

Motor unit number estimation may be a useful method to evaluate motor function recovery after spinal cord transection in rats

Experimental rat study. To investigate changes in motor unit number estimation (MUNE) value of the medial gastrocnemius (MG) muscle after thoracic spinal cord transection in rats and to correlate the MUNE with hindlimb motor function recovery. China Rehabilitation Research Center, Beijing, China. Twelve rats were subjected to spinal cord transection or sham surgery and then evaluated by MUNE and the Basso, Beattie and Bresnahan (BBB) behavioral scale 56 days after the surgery. The MUNE values of the MG muscle were significantly decreased from baseline at 14-49 days after surgery. However, they returned to near pre-injury levels after 56 days. Rats recovered progressively from the severely impaired hindlimb motor function induced by spinal cord injury, as indicated by a gradual increase in BBB score during days 3-49 after surgery. However, this behavioral recovery was only partial and reached a plateau on day 49. Finally, there was a U-shape-like correlation between changes in MUNE values and BBB scores after thoracic spinal cord transection. Time-dependent changes in the functional motor unit number may occur in spinal segments caudal to the transection level, and MUNE could be a useful method to evaluate motor function recovery.