Objective To investigate the method of cultivation and the feature of differentiation of spinal cordderived stem cells in vitro.Methods The neural stemcells from spinal cord of 15 days fetal rats were harvested and cultivated in aserumfree limited medium. The stem cells were induced to differentiate and theresults were identified by cellular immunohistochemistry. Results Lots of stem cells were obtained from the spinal cord of fetal rats and the sphere of stemcells was formed about 10 days. Neural stem cells can give rise to mature neurons and astrocytes.Conclusion Epidermal growth factor/basic fibroblast growth factor serum-free limited medium can promote the proliferation activity ofthe stem cells. Spinal cord-derived stem cells can differentiate into glial cells and neurons.
OBJECTIVE: To investigate the protective effect of tumor necrosis factor-alpha(TNF-alpha) on spinal motor neurons after peripheral nerve injury. METHODS: Twenty Wistar rats were divided into two groups, the right sciatic nerves of 20 Wistar rats were transected, the proximal stumps were inserted into a single blind silicone tube. 16 microliters of normal saline(NS) and TNF-alpha(30 U/ml) were injected into the silicone tubes. After 2 weeks, the 4th, 5th lumbar spinal cord were taken for examination. Enzyme histochemical technique and image analysis were used to show acetylcholinesterase(AChE) and nitric oxide synthase(NOS) activity of spinal motor neurons. RESULTS: The number of AChE and NOS staining neurons were 8.65 +/- 1.98 and 5.92 +/- 1.36 in the experimental group and 6.37 +/- 1.42 and 8.67 +/- 1.45 in the control group respectively, there were significant difference between the two groups(P lt; 0.01). CONCLUSION: It suggests that TNF-alpha has protective effect on motor neurons after peripheral nerve injury.
ObjectiveTo analyze the curative effect of vacuum sealing drainage for early deep infection after posterior spinal internal fixation. MethodsFrom March 2009 to March 2012, 9 patients with early deep infection after posterior spinal internal fixation, including 5 males and 4 females aged between 21 and 64 years, averaging at 44.6, underwent debridement and vacuum sealing drainage (VSD). Original fixtures in all the patients were not taken out. ResultsThe patients were treated by VSD for once to three times with an average of 2.1 times, and VSD continued for 3 to 7 days every time. Two patients underwent one time of VSD, 4 underwent twice, and 3 underwent three times. In these patients, 6 achieved wound healing after VSD, one changed to ordinary dressing and wound suturing two weeks later due to hemorrhage of VSD, one changed to ordinary dressing and wound suturing wound three weeks later due to impeded drainage, one accepted skin-grafting after three times of VSD. All the patients were followed up for 6 to 38 months (18 months on average) and all of them were cured. There was no obvious back pain or signs of incision infection. The X-ray films showed that there were no bone destruction and the white blood cell count, erythrocyte sedimentation rate, and C-reactive protein significantly decreased to normal after surgery. ConclusionOne-stage debridement and vacuum sealing drainage therapy is an effective method for treating early deep infection after posterior spinal internal fixation.
Objective To explore the influence factors for the functional improvement after the fetal olfactory ensheathing cell (OEC) transplantation for chronic spinal cord injury(SCI). Methods The olfactory bulbs were harvested and trypsinized down to single fetal OEC. They were cultured for 12-17 days prepared for use. From November 2001 to December 2003, a total of 300 patients volunteered for the fetal OEC transplantation, among whom 222 suffered from complete chronicSCI and 78 suffered from incomplete chronic SCI. The procedures were performed on the patients with a disease course ranging from 6 months to 31 years (average 3.1 years) after their injuries. The fetal OEC was transplanted by the form of injections into the spinal cord at the upper and lower ends of the injury site. All the patients were assessed by the ASIA standard before the transplantation and 2-8 weeks after the transplantation. The influence factors including age, sex, duration after the injury, and injury degreesand levels were compared with those in the functional improvement after fetal OEC transplantation. Results The partially-improved neurological functions assessed by the ASIA standard were indicated by the motor scores increasing from 39.1±20.6 to 45.9±20.3 (Plt;0.001), the light touch scores from 51.7±24.9 to 63.4±23.0 (Plt;0.001), and the pin prick scores from 53.0±24.2 to 65.3±22.7(Plt;0.001). There was no significant difference in the functional improvement of the motor, light touch, and pin brick when compared with the age, sex, duration after theinjury, and the injury degrees and levels. The motor scores and light touch scores at the cervical level were higher than the scores at the thoracic level. Conclusion The fetal OEC transplantation can partially improve the neurological functions quickly in treatment of the chronic spinal cord injury. All the influence factors except the motor scores and light touch scores, which were higher at the cervical level than at thoracic level, have no impact on the functional improvement after the fetal OEC transplantation.
In order to study the prophylactic and therapeutic effect of methyl-prednisolone (MP) on traction injury of spinal cord, 48 rabbits were divided into four groups randomly. According to decreasing amount of the amplitude of P1-wave, 50% reduction lasted for 5 min and 10 min with MP as experimental group, and 50% 5 min and 10 min with NS as control, the changes of amplitudes were monitored by, and the function of the spinal cord was assessed. The amounts of MDA and SOD of the spinal cord tissue were determined and the pathomorphological changes of the spinal cord were observed. The results showed that in the experimental groups, the recovery of P1-wave was quicker, the Tarlov and Molt value were decreased, the density of gray matter of the anterior horn and the myelinated nerve fiber of white matter of 100 microns diameter were higher, the SOD and MDA was decreased and the degenerative and necrotic degree of neuron and nerve fiber were milder. Where in the control groups all the above items were just on the opposite. The conclusions list as follows: the application of MP before operation of spinal deformity might prevent traction injury of the spinal cord during operative correction of spinal deformity, and could also minimized the secondary damage to spinal cord from traction injury if MP was used in time. The action to MP were summarized as improving the microcirculation, inhibiting the hyperoxidation of lipid and accelerating the recovery of SCEP.
OBJECTIVE: Both primary and metastatic tumor of spine can influence spinal stability, spinal cord and nerves. The principles of dealing spinal tumor are resection of tumor decompression on spinal cord and reconstruction of spinal stability. METHODS: Since Aug. 1993 to Oct. 1996, 15 cases with spinal tumor were treated, including 4 primary spinal tumor and 11 metastatic tumor. Tumor foci were mainly in thoracic and lumbar spine. Graded by Frankel classification of spinal injuries, there were 1 case of grade A, 1 of grade B, 3 of grade C, 5 of grade D and 5 of grade E. Tumors of upper lumbar spine and thoracic spine were resected through anterior approach. Posterior approach also was adopted once posterior column was affected. Tumors of lower lumbar spine were resected by two-staged operation: firstly, operation through posterior approach to reconstruct spinal stability: secondly, operation through anterior approach. After resection of tumor, the spines were fixed by Kaneda instrument, Steffee plate or Kirschner pins. To fuse the spine, bone grafting was used in benign tumor and bone cement used in malignant tumor. RESULTS: Except one patient died from arrest of bone marrow, the others were followed up for 3 to 20 months. Postoperatively, 11 patients could sit up on one foot with the help of body supporter, and 9 patients could walk in two weeks under careful monitoring. There was no exacerbation of symptom and failure of fixation. The function of spinal cord was improved: 1 case from grade B to grade E, 1 from A to C, 2 from C to E and 4 from D to E. CONCLUSION: The spine can be reconstructed for weight bearing early by internal fixation. The symptom can be relieved and the nervous function can be improved by resection of tumor and decompression.
Objective To review the advances in repair of spinal cord injury by transplantation of marrow mesenchymal stem cells(MSCs). Methods The related articles in recent years were extensively reviewed,the biological characteristic of MSCs,the experimental and clinical studies on repair of spinal cord injury by transplantation of MSCs,the machanisms of immigration and therapy and the problems were discussed and analysed. Results The experimental and clinical studies demonstrated that the great advances was made in repair of spinal cord injury by transplantation of MSCs. After transplantation, MSCs could immigrate to the position of spinal cord injury, and differentiate into nervelike cells and secrete neurotrophic factors.So it could promote repair of injuryed spinal cord and recovery of neurologicalfunction. Conclusion Transplantation of MSCs was one of effective ways in repair of spinal cord injury, but many problems remain to be resolved.
Objective To explore the clinical features of spontaneous spinal epidural hematoma (SSEH) and to find out factors influencing its prognosis. Methods From September 1998 to October 2006, 23 patients with SSEH (10 males and 13 females) were treated. Their ages ranged from 10 to 69 years. The primary neurological status were classified as grade A in 7 patients,B in 2 patients, C in 4 patients, D in 9 patients and E in 1 patients accordingto ASIA grading system. The progressive intervals of their symptoms were divided as four period: less than 12 hours (12 patients), 12 to 24 hours(2 patients), 24 to 48 hours(3 patients) and more than 48 hours(6 patients). SSEH was diagnosedby MRI or by histopathological examination. The cases history, laboratory examination, radiological image, treatment, pathological result and prognosis were recorded and analyzed after 3 month. Results In 23 patients, there were 1 case of deterioration, 8 cases of no change, 9cases of improvement and 5 cases of complete recovery. The gender had no correlationwith prognosis(P＞0.05). In the patients who had shorter progressive interval and more severe edema of spinal cord, the prognosis was worse(P＜0.05). Inthe patients who had mild neurological deficit, the prognosis was good (P＜0.01). In 17 patients undergoing surgery, the scores for prognosis was 1 point in 1 case, 2 points in 5 cases, 3 points in 6 cases and 4 points in 5 cases; the operation time had no correlation with prognosis(r=0.056, P＞0.05). In6patients undergoing conservative treatment, the scores for prognosis were 2 points and 3 points in 3 cases respectively. Conclusion Prognosis of patient with SSEH is influenced by his primary neurological status, progressive interval, spinal edema and size of hematoma. The major treatment is surgical evacuation of hematoma as early as possible to break the aggravation of spinal function. Conservative treatment is not considered unless the neurological defects recovered in the early period.
Objective To investigate the division, prol iferation and differentiation abil ities of nestin+/GFAP+cell after spinal cord injury and to identify whether it has the characteristic of neural stem cells (NSCs). Methods Twelvemale SD rats, aged 8 weeks and weighing 200-250 g, were randomized into 2 groups (n=6 per group): model group inwhich the spinal cord injury model was establ ished by aneurysm cl ip compression method, and control group in which no processing was conducted. At 5 days after model ing, T8 spinal cord segment of rats in each group were obtained and the gray and the white substance of spinal cord outside the ependymal region around central tube were isolated to prepare single cellsuspension. Serum-free NSCs culture medium was adopted to culture and serum NSCs culture medium was appl ied to induce differentiation. Immunohistochemistry detection and flow cytometry were appl ied to observe and analyze the type of cells and their capabil ity of division, prol iferation and differentiation. Results At 3-7 days after injury, the model group witnessed a plenty of nestin+/GFAP+ cells in the single cell suspension, while the control group witnessed few. Cell count of the model and the control group was 5.15 ± 0.71 and 1.12 ± 0.38, respectively, indicating there was a significant difference between two groups (P lt; 0.01). Concerning cell cycle, the proportion of S-phase cell and prol iferation index of the model group (15.49% ± 3.04%, 15.88% ± 2.56%) were obviously higher than those of the control group (5.84% ± 0.28%, 6.47% ± 0.61%), indicating there were significant differences between two groups (P lt; 0.01). In the model group, primary cells gradually formed threedimensional cell clone spheres, which were small in size, smooth in margin, protruding in center and positive for nestin immunofluorescence staining, and large amounts of cell clone spheres were harvested after multi ple passages. While in the control group, no obvious cell clone spheres was observed in the primary and passage culture of single cell suspension. At 5 days after induced differentiation of cloned spheres in the model group, immunofluorescence staining showed there were a number of galactocerebroside (GaLC) -nestin+ cells; at 5-7 days, there were abundance of β-tubul in III-nestin+ and GFAP-nestin+ cells; and at 5-14 days, GaLC+ ol igodendrocyte, β-tubul in II+ neuron and GalC+ cell body and protruding were observed. Conclusion Nestin+/GFAP+ cells obtained by isolating the gray and the white substance of spinal cord outside the ependymal region around central tube after compressive spinal cord injury in adult rat has the abil ity of self-renewal and the potential of multi-polarization and may be a renewable source of NSCs in the central nervous system.
Objective To discuss the clinical value of whole spine magnetic resonance imaging (WSMRI) in practice of neurosurgical spinal surgery. Method A total of 70 cases examined using WSMRI between January 2015 and December 2016 were collected and analyzed retrospectively. Results All patients got clear images of WSMRI. Eighteen cases got important information, including spinal variation (1 case), multiple lesions (3 cases), combined lesions (6 cases) and large range multi-segmental lesions (8 cases), which were missed by single-segment MRI . Conclusions WSMRI can show all the spine, spinal cord and surrounding tissue in one image at one time. It has high clinical value because of its accurate positioning, comprehensiveness, time saving, and low rate of misdiagnosis and missed diagnosis.