| Part 1 The research of umbilical cord mesenchymal stem cellsdifferentiate into nerve cells and repair of spinal cord injuryObjective: As a spinal disease induced by trauma,spinal cord injury(SCI)with the characteristic of sensory,motor and autonomic nerve function defect below the damage section.The epidemiologic survey displayed that 130 thousand patients occurred spinal cord injury in each year,and more than 2.5 million patients were plagued by various sequelae induced by spinal cord injury.More than $6 billion a year was spend by the SCI patients and caused a heavy burden to family and society.At present,the clinically treatment of SCI was mainly limited at medicine,surgery,and various physical intervention.Stem cell therapy was still stay in animal related research level.Such as marrow stromal stem cells transplantation,olfactory ensheathing cells transplantation,umbilical cord blood mesenchymal stem cell transplantation and mesenchymal stem cells(MSCs)transplantation source of umbilical cord huatong glue.Due to the extensive sources,easy expansion,no restrictions for the ethics,umbilical cord mesenchymal stem cells(UCMSCs)had become a new focus and direction in the basic experiments for the treatment of SCI.Under certain environment inducement,UCMSCs could differentiate into different functional cells and promoted the protective factors generation.When the UCMSCs differentiated into nerve cells,it could promote the restoration of nerve tissue and improve the neurological function of SCI rat.Some domestic scholars adopt the method of transplantation of human umbilical cord blood stem cells(hUCMSCs)to treat SCI rats.Although obtained a satisfactory curative effect,but there were low efficiency in stem cells differentiation.In this study,we intend to improve the differentiation rate and transplanting it to the SCI mice,in order to provide a theoretical basis in treating this intractable disease.Methods: Collected the umbilical cord and separated human umbilical cord mesenchymal stem cells by explants method,collagenase digestion method and collagenase with pancreatic enzyme digestion method.After the training and amplification,hUCMSCs were transfect by the Nurr-1 gene and differentiation into neural cells combined with cocktail factors(ATRA,GGF-2,bFGF,PDGF and forskolin).Then immunofluorescence technique was used to detect the markers of nerve cells.Establishment of spinal cord injury model by kunming mice,and then transplanted the cells to the spinal cord.Immunohistochemical was done to detect engraftment situation and observe the therapeutic effect.Results: After induction,the hUCMSCs turned round and shrink and then become more angle sample after stretching.In the growing process,the cell body gradually appeared synaptic structure.Two weeks after induction,nerve cells specific markers of MAP-2 and MBP appeared.The over expressed umbilical cord mesenchymal stem cells had the highest efficiency to differentiate into nerve cells.The SCI model mice had a serious damage with neuronal death in the gray matter,anterior horn of neurons partly surviving and some of neuron appeared nuclear pyknosis.The cells propagation in mass quantity at the injured area after transplanted UCMSCs,anti-human nucleus of positive cells could be detected by fluorescence microscope.After the transplantation,the Basso,Beattie& Bresnahan locamotor rating scale(BBB scale)of the experiment mice had a significant increase,and the hindlimb motor function gradually improved.Conclusion: Under the Nurr-1 gene transfection and cocktail factors inducement,UCMSCs could get the best differentiation and proliferation.If transplanted them to the injury area of the spinal cord,the hind limb function of experiment mice could significantly be improved.It showed a definite effect in treating spinal cord injury.Part 2 Comparison of clinical curative for cervical spinal cord injurywithout fracture or dislocation by surgical and non-surgicaltreatmentObjective: As a special type of spinal cord injury,cervical spinal cord injury without fracture or dislocation(CSCIWFD)usually suffered a mild damage at the cervical region and without obvious cervical spine fracture or dislocation at the imageological examination.But it had a relative seriously symptoms.This kind of injury accounted for 23% of the total number of cervical spinal cord injury.The treatment of CSCIWFD was mainly divided into 2 types.One was to use high-dose glucocorticoids for impacting,quiescent neurotrophic drugs and gave hyperbaric oxygen treatment(non-surgical therapy).The other way was to give a sufficient spinal cord decompression surgery.Each treatment could obtain a positive degree of neurological function recovery.However,as an invasive operations,if the overall curative effect was better than that of non-surgical treatment? Aiming at this problem,we conducted the clinical comparative study.Methods: A retrospective study was performed in 110 patients who were diagnosed as CSCIWFD at outpatient and in hospitalization.Among of them,92 patients accepted for more than 18 months of clinic follow-up.Based on the treatment,they were divided into 2 groups,38 cases in group A(non-surgical group).There were 23 male and 15 female patients ranging from 19-63 years(mean,42.1 years).Fifty-four cases in group B(surgical group).There were 35 male and 19 female patients ranging from 17-62 years(mean,40.8 years).The operative time,the intraoperative blood loss and complications were record in the surgical group.The neurological improvement based on JOA score and ASIA classification standard was recorded and compared at the point of pre-operation,1 week after treatment,1 month after treatment and at the last follow-up.Results: All patients were operated successfully in group B,and 10 cases accepted anterior cervical discectomy and internal fixation,17 cases accepted anterior cervical corpectomy and internal fixation,16 cases accepted posterior laminoplasty combined with titanium plate fixation,11 cases accepted posterior laminectomy and lateral mass screw fixation.The average operative time was 175.2 min and the mean blood loss was 210.6 ml.In group A the JOA score was 7.0±1.8,7.9±2.0,9.1±2.2,9.9±2.3,10.5±2.4,11.2±2.9 at the point of pre-operation,1 week after treatment,1 month after treatment,3 month after treatment,6 months after treatment and at the last follow-up,which there was significant difference compared with the pre-operation(P<0.05).In group B,there was significant difference when compared with the pre-operation(P<0.05).Besides,the JOA score in group B was superior than group A at each point after the treatment(P<0.05).Based on ASIA degree system at last follow-up,there was A in 3 case,B in 7 cases,C in 10 cases,D in 14 cases,and E in 4 cases in the group A,which was significantly different when compared with former level(P<0.05).There was A in 3 case,B in 6 cases,C in 16 cases,D in 17 cases,and E in 12 cases in the group B,which was significantly different when compared with former level(P< 0.05).During the treatment period,1 case occurred alimentary tract hemorrhage,2 cases occurred pulmonary infection,2 cases pressure ulcer and 1 case deep vein thrombosis in group A.One case occurred wound infection,2 cases occurred extradural hematoma,1 cases pressure ulcer and 1 case cerebrospinal leak in group B.There was no significant difference between the 2 group(P>0.05).Conclusion: When compared with the non-surgical group,the procedure of spinal cord decompression and internal fixation has the characteristic of promote the neurological function more superior and faster in the treatment of cervical spinal cord injury without fracture or dislocation patient.However,the surgical treatment does not increase the incidence of perioperative complications.Part 3 Curative effect analysis of anterior or posterior decompressionwith internal fixation to treat cervical spinal cord injury withoutfracture or dislocationObjective: It had been confirmed that surgical procedure had a better therapeutic effective than all kinds of non-surgical therapy in the treatment of cervical spinal cord injury without fracture or dislocation(CSCIWFD).So once a CSCIWFD patient with a certain degrees of neurological defect appeared in the hospital,a positive and appropriate surgical procedure should be done to restore the vertebral canal unobstructed.There were so many surgical methods that used to treat CSCIWFD.Such as anterior cervical discectomy and internal fixation,anterior cervical corpectomy and internal fixation,posterior laminoplasty combined with titanium plate fixation and posterior laminectomy and lateral mass screw fixation.According to the surgical approach,they were divided into the anterior and posterior spinal cord decompression.Whether the anterior and posterior approach had any differences in the clinical effective? In this study we try to solve the problem.Methods: A retrospective study was performed in 82 patients who were diagnosed as CSCIWFD in our hospitalization.Based on the treatment,they were divided into 2 groups,45 cases in group A(anterior group).There were 26 male and 19 female patients ranging from 22-62 years(mean,41.6 years).They accepted the anterior decompression and internal fixation.Thirty-seven cases in group B(posterior group).There were 24 male and 13 female patients ranging from 19-64 years(mean,42.9 years).The operative time,the intra-operative blood loss and complications were record in the 2 group.The neurological improvement based on JOA score and ASIA classification standard was recorded and compared at the last follow-up.Results: All patients were operated successfully.In group A,the average operative time was 62.6 min and the mean blood loss was 105.1 ml.In group B,the average operative time was 117.9 min and the mean blood loss was 240.8 ml,there was significant difference between the 2 groups(P<0.05).In group A,the JOA score was 6.8±1.5,9.5±1.9,11.6±2.6,13.5±3.1 at the point of pre-operation,2 week after treatment,3 month after treatment and at the last follow-up,which there was significant difference compared with the pre-operation(P<0.05).In group B,the JOA score was 6.6±1.4,8.6±1.7,10.9±2.5,13.3±3.4 at the point of pre-operation,2 week after treatment,3 month after treatment and at the last follow-up,which there was significant difference compared with the pre-operation(P<0.05).At the point of 2 weeks,there was significant difference between them(P<0.05).But there was no significant difference at the point of 3 month after treatment and at the last follow-up(P>0.05).Based on ASIA degree system at last follow-up,there was A in 1 case,B in 6 cases,C in 11 cases,D in 18 cases,and E in 9 cases in the group A,which was significantly different when compared with former level(P<0.05).There was A in 1 case,B in 4 cases,C in 11 cases,D in 13 cases,and E in 8 cases in the group B,which there was significantly different when compared with former level(P<0.05).During the hospitalized period,2 case occurred cerebrospinal fluid leakage,1 case recurrent laryngeal nerve injury and 1 cases of titanium mesh subsidence in group A.One case occurred wound infection,1 case liquefactive necrosis and 2 cases of extradural hematoma in group B.There was no significant difference between the 2 group(P>0.05).Conclusion: Both the anterior and posterior spinal cord decompression has an effective curative in treating cervical spinal cord injury without fracture or dislocation,and the incidence of perioperative complications are basically the same.But the anterior approach has the characteristic of with small surgical trauma,shorter operation time,less intraoperative bleeding.So,we should chose a reasonable operative procedure based on the compressed segments,oppression source and the length of intramedullary high signal. |
PDF Full Text Request