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BACKGROUND:Endothelin has been found to be involved in the breakdown of the blood-spinal cord barrier after spinal cord injury,and stem cell-derived exosomes can reduce the permeability of the blood-spinal cord barrier and repair spinal cord injury. OBJECTIVE:To investigate whether exosomes produced by human umbilical cord mesenchymal stem cells can reduce the permeability of the blood-spinal cord barrier by inhibiting endothelin-1 expression,thus repairing spinal cord injury. METHODS:Exosomes were extracted from the cultured supernatant by the hyperspeed centrifugation method.The morphology of exosomes was observed by transmission electron microscope.The expression levels of tsg101 and CD63 were detected by western blot assay.Eighty SD rats were randomly divided into sham operation group,model group,exosome group,and endothelin-1 group(n=20).The modified Allen's method was used to create the rat model of spinal cord injury.In the endothelin-1 group,10 μL(1 μg/mL)endothelin-1 was injected directly into the injured area with a microsyringe.Immediately,1 day,2 days after operation,sham operation group and model group were injected with 200 μL PBS solution through the tail vein;the exosome group and endothelin-1 group were injected with 200 μL exosome(200 μg/mL)solution through the tail vein,respectively.Hind limb motor function scores were performed on days 1,3,7,14 and 21 after spinal cord injury.The blood-spinal cord barrier permeability was observed by Evans blue staining on day 7 after injury.The expression levels of tight junction proteins β-Catenin,ZO-1,Occludin and endothelin-1 in the spinal cord were detected by western blot assay. RESULTS AND CONCLUSION:(1)Basso-Beattie-Bresnahan score in the exosome group was significantly higher than that in the model group at 3-21 days after injury(P<0.05).Hematoxylin-eosin staining showed that spinal cord injury was greatly reduced in the exosome group compared with the model group.Basso-Beattie-Bresnahan score in the endothelin-1 group was significantly decreased compared with the exosome group(P<0.05).Spinal cord injury was more severe in the endothelin-1 group than that in the exosome group.(2)The expression of endothelin-1 in the model group was significantly increased compared with the sham operation group(P<0.05),and the expression of endothelin-1 in the exosome group was significantly decreased compared with the model group(P<0.05).(3)The blood-spinal cord barrier Evans blue exudate in the exosome group was significantly decreased compared with the model group(P<0.05).The expression levels of the tight junction proteins β-Catenin,Occludin and ZO-1 in the exosome group were increased(P<0.05);the Evans blue exudate in the endothelin-1 group was significantly increased compared with the exosome group(P<0.05).The expression level of tight junction protein was significantly decreased compared with the exosome group(P<0.05).(4)The results show that human umbilical cord mesenchymal cell-derived exosomes protect the permeability of the blood-spinal cord barrier by down-regulating the expression of endothelin-1 and play a role in the repair of spinal cord injury.
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Objective:To compare perioperative differences between posterior lumbar interbody fusion(PLIF)by a posterior median approach or open transforaminal lumbar interbody fusion(TLIF)and minimally invasive surgery TLIF(MIS-TLIF)by a paravertebral approach for lumbar spondylolisthesis(Ⅰ~Ⅱ°)in elderly patients.Methods:A retrospective study was conducted to analyze clinical data of patients aged 60 years and over with lumbar spondylolisthesis(n=68)who underwent PLIF or MIS-TLIF from January 2014 to December 2017.Patients were divided into the PLIF group(n=40, 12 males and 28 females)and the MIS-TLIF group(n=28, 6 males and 22 females)according to the type of surgery.Differences in operating time, blood loss, exposure frequency, postoperative drainage volume, indwelling duration, hospital stay length and complications were compared between the groups.Results:There was no significant difference in operative time between the PLIF group and the MIS-TLIF group(156±51)min vs.(153±38)min( P=0.77). There were significant differences in intraoperative blood loss and radiation exposure frequency between the PLIF group and the MIS-TLIF group(458±272)ml vs.(157±104)ml( P<0.001); (6.7±1.5)times vs.(30.6±6.9)times( P<0.01). There was no significant difference in intraoperative complications between the PLIF group and the MIS-TLIF group(25% or 10/40 vs.7.1% or 2/28, P=0.057). There were more postoperative drainage, longer indwelling time and hospitalization in the PLIF group than in the MIS-TLIF group(664±351)ml vs.(210±127)ml( P<0.001); (3.7±2.9)d vs.(2.2±0.8)d( P=0.002); (9.2±3.6)d vs.(6.9±1.7)d( P=0.001). The incidence of postoperative neurological complications was not significantly different between the PLIF group and the MIS-TLIF group(7.5% or 3/40 vs.14.3% or 4/28, P=0.365). The incidence of postoperative non-neurological complications was higher in the PLIF group than in the MIS-TLIF group(27.5% or 11/40 vs.7.1% or 2/28, P=0.036). Conclusions:Compared with PLIF, MIS-TLIF has the advantages of less intraoperative bleeding and postoperative drainage, shorter indwelling duration of urinary catheters, shorter hospital stays and fewer complications of non-neurological complications in treating Ⅰ-Ⅱ degree lumbar spondylolisthesis in the elderly, but it requires more radiation exposure.
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Cervical spondylotic myelopathy is a common and frequently-occurring disease in Orthopaedics, especially with multi-segmental cervical spondylotic myelopathy. There are several pathogenic factors for cervical spondylotic myelopathy. The clinical symptoms are serious, and the imaging manifestations are complex. Most of them are accompanied by serious neurological damage, which seriously affects the quality of life of patients. Furthermore, some patients have serious cervical spinal cord injury symptoms, which endanger their lives after mild trauma. Therefore, early diagnosis, early treatment and surgery are the most effective methods at present, which could effectively eliminate the factors of spinal cord compression and reconstruct the stability of cervical spine function. However, the choice of surgical approach and decompression fusion has always been the focus of debate. The primary purpose of surgery for cervical spondylotic myelopathy is to completely relieve the compression of spinal cord. According to the imaging characteristics of cervical spondylotic myelopathy, it is the key to the success of the operation to determine the decompression and reconstruction of cervical spondylotic myelopathy. Anterior approach can be adopted in patients with the compression factors located in front of the spinal cord, including disc protrusion, mild osteophyte or local ossification posterior longitudinal ligament, less lesion segments, small compression range, cervical kyphosis or local kyphosis deformity, easy resection of compression and sagittal imbalance correction. Decompression fusion can be selected through intervertebral space, subtotal vertebral body resection and mixed decompression for fusion and fixation. In order to retain the active function of cervical vertebrae, artificial disc replacement and fusion can be selected. For patients with the compression factors located in the posterior part of the spinal cord, including ligamentumflavum hypertrophy or ossification, congenital developmental spinal canal stenosis, there are many factors causing compression in front of the spinal cord. The range is larger, the lesion segment is longer. Thus, it is difficult to resect thoroughly in front of the spinal cord with higher risk of anterior surgery. Although the posterior approach is indirect decompression with the help of posterior spinal cord movement, it plays a good role in improving nerve function with fewer complications and relatively higher safety. There are absolute surgical indications and relative indications for two approaches. Combining anterior and posterior surgery has advantages of complete decompression and strong fixation. However, it has the advantages of great trauma and high risk, so it is necessary to apply it in clinical practice. Moreover, no matter how to choose the approach, we should use perfect imaging data combined with clinical symptoms and signs to clarify the pathological factors of spinal cord com pression, the severity of spinal cord injury, the scope of compression and the responsible segment, and formulate an individualized operation plan.
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Objective To investigate the application value of single or combined detection of HE4 and CA125 in the differentia‐tion diagnosis of endometriosis .Methods The detection results of serum HE4 and CA125 levels in 105 cases of patients with ovari‐an endometriosis (endometriosis group) ,53 cases of patients with (ovarian malignant group) tumor and 64 healthy women undergo‐ing the healthy physical examination(control group) were performed the retrospective and statistical analysis .Results The positive rates of serum HE4 in the endometriosis group ,ovarian malignant tumor group and control group were 57 .1% ,90 .6% and 12 .5%respectively with statistically significant difference among them (P< 0 .05) .The positive rates of serum CA125 in the above 3 groups were 44 .8% ,92 .5% and 14 .0% respectively ,and the differences were statistically significant(P<0 .05) .(3)The sensitivity and specificity of 70 pmol/L≤ HE4<150 pmol/L for diagnosing endometriosis were 57 .14% and 53 .86% respectively ,which of 35 U/mL≤CA125<150 U/mL for diagnosing endometriosis were 44 .76% and 61 .24% .But the sensitivity and specificity of their combined detection were 70 .32% and 60 .38% .There were statistically significant differences of sensitivity and specificity among single and combined detecion of HE4 and CA125(P<0 .05) .Conclusion Serum HE4 and CA125 levels are significantly elevated in the patients with endometriosis ,their combined detection could further raise the sensitivity for diagnosing endometriosis .
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Objective To investigate the selection of surgery and clinical outcomes of upper cervical injuries.Methods 25 upper cervical injury patients were involved in this retrospective study from November 2011 to June 2014.Including 20 males and 5 females with mean age of 37.1 years old (range,14-55 years old).Individual operation methods were based on the comprehensive evaluation of specific situations including the clinical manifestation,the type of the injuries and the imaging data.HaloVest distraction was applicated before operation.The surgery by anterior approach were performed for 7 patients and posterior approach were performed for 18 patients.Preoperative and postoperative American Spinal Injury Association (ASIA) grade and Functional Independence Measurement (FIM) score were studied to evaluate the nerve functional restoration.Imaging data before and after the operation were contrasted to evaluate the reduction of the fracture,the bone union,the fusion of the bone graft and the condition of the internal fixation.Wilcoxon Singed Rank Test was applied to compare the FIM score between pre-operation and last follow-up.Results 15 patients presented neurological function deficit because of cervical spinal cord compromise.All cases were followed up for 6-35 months (mean 18.2 months),showing good clinical and radiological effects.Solid fusion was obtained in all patients among 3-12 months.The ASIA grade improved by an average of 1.1 (6 months after operation) and 1.2 (12 months after operation).There was significant difference in FIM score between pre-operation and last follow-up.One patient got cerebrospinal fluid leakage.Conservative treatment was implemented with the Trendelenburg position,rehydration fluids and so on.Removal of drainage tube 8 days later when the drainage was less than 30 ml/24 h.No incision infection,cerebrospinal fluid leakage,migration or breakage of internal fixation was observed at the last follow-up.Conclusion The type of upper cervical injuries are complicated,the characteristics of fracture,dislocation and nerve injury in different patients are different.The specific situation should be evaluated comprehensively to make individual operation methods.The success of the operation requires the proficiency of the anatomic basis,the biomechanical characteristics,precise entrance point and direction in operation,appropriate diameter of the screw and suitable depth of the screw road.
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BACKGROUND:A large number of studies have shown that adult stem cels derived from multiple tissues are available to differentiate towards nucleus pulposus-like celsin vitro. It is unclear whether mesenchymal stem cels derived from nucleus pulposus tissues have the ability to differentiate towards nucleus pulposus-like phenotypes induced by transforming growth factor-beta 1. Up to now, there are few reports on the difference between the differentiation ability of mesenchymal stem cels derived from nucleus pulposus tissues and adipose-derived mesenchymal stem cels. OBJECTIVE:To compare the ability of mesenchymal stem cels derived from nucleus pulposus tissues and adipose-derived mesenchymal stem cels differentiating into nucleus pulposus-like cels under induction of transforming growth factor-beta 1. METHODS:The groin fat tissue and the coccygeal spine of rats were taken respectively to isolate and culture mesenchymal stem cels derived from nucleus pulposus tissues and adipose-derived mesenchymal stem cels by mechanical enzyme digestion method. Flow cytometry was employed to detect the expression of CD105, CD90, CD29, CD45, CD44, CD34, and CD24 of both two kinds of stem cels. Mesenchymal stem cels derived from nucleus pulposus tissues and adipose-derived mesenchymal stem cels were divided into complete induction group (complete induction medium with transforming growth factor-beta 1), incomplete induction group (complete induction medium without transforming growth factor-beta 1) and control group(DMEM/F12 containing 10% fetal bovine serum and 100 mg/L penicilin/streptomycin), respectively. After 14 days of culture, real-time PCR was used to detect the expression of colagen type II, Aggrecan and SOX-9 in each group. RESULTS AND CONCLUSION:CD105, CD90, CD29 expressed positively and CD45, CD44, CD34, CD24 negatively in both two kinds of stem cels. After 14 days of induced differentiation, the expressions of colagen type II, Aggrecan and SOX-9 in the two kinds of cels were significantly higher in the complete induction groups than in the control groups (P < 0.05). Under the induction of transforming growth factor-beta 1, the expression of colagen type II, Aggrecan and SOX-9 in mesenchymal stem cels derived from nucleus pulposus tissues was significantly higher than that in adipose-derived mesenchymal stem cels (P < 0.05). These findings suggest that both two kinds of mesenchymal stem cels have the ability to differentiate towards nucleus pulposus-like cels induced by transforming growth factor-beta, and mesenchymal stem cels derived from nucleus pulposus tissues may be more suitable as seed cels for nucleus pulposus tissue engineering research.
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<p><b>BACKGROUND</b>Recent studies have indicated that human nucleus pulposus contain mesenchymal stem cells (NP-MSCs). However, the immunophenotypic variation of NP-MSCs in vitro was unclear. The present study was conducted to address the immunophenotypic variation of mesenchymal stem cells in nucleus pulposus under continuous proliferation in vitro and show the difference between mesenchymal stem cells and nucleus pulposus cell.</p><p><b>METHODS</b>Tissue samples were obtained from thoracolumbar burst fracture patients and degenerative disc disease patients who underwent discectomy and fusion procedures. Flow cytometric and laser scanning confocal microscope (LSCM) were used to detect the variation of mesenchymal stem cells in nucleus pulposus which were expressing CD105 and CD24 in condition with or without transforming growth factor β1 (TGF-β1).</p><p><b>RESULTS</b>More than 90% of the analyzed primary cells of mesenchymal stem cells in nucleus pulposus fulfilled the general immunophenotyping criteria for MSCs, such as CD44, CD105 and CD29, but the marker of mature NP cells characterized as CD24 was negative. In continuous cultures, the proportion of mesenchymal stem cells which were expressing CD44, CD105 and CD29 in nucleus pulposus gradually decreased. The mesenchymal stem cells in nucleus pulposus cells were positive for CD105 and CD29, with slight positivity for CD44. The CD24 expression gradually increased in proliferation. Biparametric flow cytometry and laser scanning confocal microscopy confirmed the presence of cells which were expressing CD105 and CD24 independently, and only a small part of cells expressed both CD105 and CD24 simultaneously. TGF-β1 could stimulate mesenchymal stem cells in nucleus pulposus to express CD24.</p><p><b>CONCLUSIONS</b>Non-degenerative and degenerative NP contains mesechymal stem cells. The variation of CD24 can be used as a marker to identify the NP-MSCs differentiation into NP-like cells.</p>
Subject(s)
Adult , Female , Humans , Male , Young Adult , Antigens, CD , Metabolism , CD24 Antigen , Metabolism , Cell Differentiation , Physiology , Cells, Cultured , Endoglin , Flow Cytometry , Hyaluronan Receptors , Metabolism , Integrin beta1 , Metabolism , Intervertebral Disc , Cell Biology , Metabolism , Mesenchymal Stem Cells , Cell Biology , Metabolism , Receptors, Cell Surface , MetabolismABSTRACT
BACKGROUND:Endogenous stem cells have no repair effects on the process of disc degeneration. Authors assumed that this maybe associate with abnormal effects of related etiological factor, resulting in an inhibitory effect on the function of nucleus pulposus-derived mesenchymal stem cells. OBJECTIVE:To investigate the effects of inflammatory cytokine interleukin-1βon biological characteristics of nucleus pulposus-derived mesenchymal stem cells of rats. METHODS:Lumbar spinal nucleus pulposus was obtained from 3-month-old male Sprague-Dawley rats. Nucleus pulposus-derived mesenchymal stem cells were isolated and cultured with col agenase and sequential trypsin digestion. The expression of CD24, CD34, CD45, CD90 and CD105 was detected using flow cytometry. Stem cellgene SOX2 and Nanog expression was measured using RT-PCR. Adipogenic, osteogenic and chondrogenic abilities of nucleus pulposus-derived mesenchymal stem cells were observed. The apoptotic rate of interleukin-1β-treated nucleus pulposus-derived mesenchymal stem cells was detected using flow cytometry. Fluorescent quantitative PCR was used to measure the expression of SOX9, proteoglycan, type II col agenase and caspase-3 gene after nucleus pulposus-derived mesenchymal stem cells were treated with interleukin-1β.
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Objective To investigate the clinical efficacy,operative essentials and indications of hybrid spinal fusion surgery for cervical spondylotic myelopathy.Methods From August 2008 to December 2011,thirty-eight patients with cervical spondylotic myelopathy underwent hybrid spinal fusion surgery in our hospital.There were 27 males and 11 females,aged from 33 to 70 years (average,51 years).A total of 86 segments were treated (fusion 48 vs.non-fusion 38).Twenty-eight patients underwent a two-level surgery,and ten patients received a three-level surgery.The Japanese Orthopaedic Association (JOA) score and Visual analogue scale (VAS) were used to evaluate pre-and post-operative neurological function and pain,respectively.The pre-and post-operative range of motion of the cervical spine was measured according to Xrays.Moreover,the surgical complications were recorded and analyzed.Results Thirty-seven patients were followed up for 15 to 55 months (average,29.1 months).The improvement of neurological function was obtained in 36 patients.The JOA score was improved from preoperative 10.5±1.57 to 14.3±1.97 at final follow-up,with an improvement rate of 58.46％,and the results were excellent in 16 cases,fair in 20 cases and poor in 1 case.The VAS was improved from preoperative 7.3±1.04 to 3.2±1.41 at final follow-up.The Cobb angle changed from preoperative 25°±3.21°to 20°±2.56°at final follow-up.After operation,the neurological function was not restored in 1 case; hoarseness and bucking occurred in 2 cases; sore throat occurred in 22 cases; anterior displacement of prosthesis (PCM) occurred in 3 cases.Other patients had no complications,such as displacement,loosening and heterotopic ossification.Conclusion In hybrid spinal fusion surgery,the lesions segments are decompressed fully,the severely degenerative segments are fused,and the motion of the non-fusion segments is reserved.As a result,not only the stability of the cervical spine is achieved,but also an obvious improvement of symptoms and a satisfactory short-term efficacy can be obtained.Therefore,this method is an alternative procedure for cervical spondylosis myelopathy.
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Objective To differential the biologic characteristics and multiple differentiation potential of mesenchymal stem cells in nucleus pulposus in (NP-MSC) scoliosis patient and patient with degenerative interverthral disc.Methods The human nucleus pulposus-mesenchymal stem cells were isolated and cultured with enzyme digestion from 2 patients of scoliosis and 2 patient with degenerative intervertbral disc separately.Cellular proliferation was detected with MTT assay and trypan blue.The immunophenotype expression of NP-MSC was detected by flow cytometry in scoliosis and degenerative group.The multiple differentiation ability of cells was assessed respectively using alizarin red dye,Oil red O dye and immunohistochemical staining.Results The primary cell morphology of scoliosis NP-MSC was the shape of spindle,while the degenerative NP-MSC was inhomogeneous.However,both of them became spindle shape after passages.The scoliosis NP-MSC was stronger than degenerative one in metabolic activity and proliferation.The percentage of positive antigen expression of CD44,CD105 and CD29 was 97％-100％ in scoliosis NP-MSC group,but 88.7％-97％ in the degenerative group.The expression of mature cell marker CD24 was negative in both groups.Furthermore,the MSC i.isolated from both groups differentiated along the osteogenic,chondrogenic but not adipogenic lineages.Conclusion The scoliosis and degenerative NP contains mesenchymal stem cells.Moreover the scoliosis NP-MSC had stronger ability of cell metabolic activity and proliferation.These cells have multiple differentiation potential with the exception of their adipogenic differentiation ability.
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[Objective]To evaluate the effectiveness of posterior lumbar interbody fusion(PLIF) in the treatment of upper lumbar disc herniation.[Method]Twenty six patients of upper lumbar disc herniation underwent posterior lumbar interbody fusion,lumbago VAS and the pain in waist and lower extremities grades were tested before operation,all patients had an average follow-up period of 20.8 months,then lumbago VAS and the pain in waist and lower extremities grades were tested at last follow-up,lumbago VAS and the pain in waist and lower extremities grades were compared through paired t-test.[Result]All 26 patients had a followed-up.Postoperative VAS had significant lower than preoperative,statistical analysis has remarkable significance(P