BACKGROUND: Growth differentiation factor 5 (GDF-5) is an important factor to regulate the formation and development of the cartilage and bone, it plays a crucial role on the promotion of repairing bone, cartilage and tendon ligament injury. OBJECTIVE: To transfect eukaryotic expression plasmid pcDNA 3.1(+)/GDF-5 to bone marrow stroma stem cells of mouse and to check the expression of extracellular matrix and proteoglycan which relates with the cartilage formation and differentiation. DESIGN, TIME AND SETTING: An in vitro observation regarding cells was performed in the central laboratory of Wuhan Union Hospital between March and December in 2008.MATERIALS: Twenty Kunming specimen male mice were offered by Experimental Animal Center of Tongji Medical College of Huazhong University of Science and Technology. The eukaryotic expression plasmid pcDNA 3.1(+)/GDF-5 was preserved at the laboratory.METHODS: The man-ow stroma stem cells were isolated from mouse bone marrow and cultured in vitro with whole bone marrow adherence method. Passage 3 cells were incubated on 6-well plate and began to transfect when they were 90% confluent. Experiment was assigned into three groups: trensfection group underwent transient transfection of liposome-mediated pcDNA 3.1(+)/GDF-5 using LipofectamineTM2000; blank plasmid group was transfected with blank plasmid pcDNA 3.1(+); control group was added with equal volume of liposome and other protocols were the same as above. MAIN OUTCOME MEASURES: The transfection efficacy was identified success by the expression of GDF-5 gene and protein using RT-PCR and immunocytochemistry at 72 heurs following transfection, cartilage matrix Ⅱ collagen expression was determined as above methods. Then marrow stroma stem cells were cultured for additional two weeks to check expression of proteoglycan with alcian blue staining.RESULTS: In the transfection group, a 219-bp specific amplification band was visible, there were brown positive stain in the cytoplasm of marrow stroma stem calls; In blank plasmid group and control group, no GDF-5 trensfection, specific amplification band or obvious stain of cytoplasm was observed. In the transfection group, the collagen Ⅱ gene was detected to express at 225 bp, with brown yellow stain in cytoplasm; in the blank plasmid group and control group, no collagen Ⅱ gene expression or SP ～ stain was observed. Alcian blue staining results showed the transfected cells were stained blue while those in the blank plasmid group and control group were not metachromasia stained.CONCLUSION: Gene trensfection of pcDNA 3.1 (+)/GDF-5 to marrow stroma stem cells can significantly raise expression of collagen II and proteoglycan, and promote the chondrogenic differentiation of marrow stroma stem cells.

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Objective To investigate the clinical efficacy of modified open-door laminoplasty with preservation ofthe unilateral paraspinal muscle ligament complex in treating ossification of posterior longitudinal ligament (OPLL) of the cervical spine.Methods From June 2015 to July 2017,thirty-two patients with OPLL of the cervical spine who underwent modified open-door laminoplasty with preservation of the unilateral paraspinal muscle ligament complex were retrospectively analyzed(modified group).There were 20 males and 12 females with the average age of 61.72±8.41 years (range 46-75 years).The decompression segment range included three cases of C2-C6,seven cases of C2-C7,eight cases of C3-C6,and fourteen cases of C3-C7.Moreover,twenty-three patients with OPLL of the cervical spine who underwent traditional unilateral open-door laminoplasty at the same time were included as controls (control group).The demographics,operation duration,and blood loss volume was recorded.The visual analogue scale (VAS),Japanese Orthopaedic Association scores (JOA),improvement rate of JOA score and neck disability index (NDI) were recorded and analyzed at preoperation,three months after operation,six months after operation and at the final follow-up.The cervical curvature,cervical curvature index (CCI),range of motion (ROM) were measured and statistically analyzed on the lateral X-ray film of the cervical spine.The union rate of the lamina hinge and that of spinous process-lamina was measured on the cross section of the CT scan.Bilateral cervical posterior muscle volume was recorded and analyzed at cross-section plane on MRI.Axial symptoms,C5 nerve root palsy and other complications were also recorded.Results All the patients were followed-up for 6 to 18 months (mean,13.04±4.67 months in control group,11.81±4.93 months in modified group).At the final follow-up,the JOA,VAS and NDI scores improved from preoperatively 6.09±0.79 to 13.43± 1.53,4.22±1.04 to 2.83±1.15 and 25.48％±3.20％ to 8.83％±5.41％ in the control group,versus 6.28±1.05 to 13.88±1.48,4.09± 1.00 to 1.16±0.57 and 25.06％±3.24％ to 5.66％±2.46％ in the modified group.The postoperative JOA score and recovery rate of the two groups was not significantly different,whereas the VAS and NDI scores were significantly reduced in the modified group compared to the control group.The maximum cervical flexion angle,cervical curvature,ROM and CCI of control group were 6.26°± 3.31°,30.17°±4.56°,11.39°±1.95° and 9.74％±4.05％ at the final follow-up,which were reduced significantly compared to pre-operation.The maximum cervical flexion angle,cervical curvature,ROM and CCI of modified group were 10.06°± 1.93°,35.03°± 5.01°,17.03°±2.86° and 14.22％±5.00％,and there were no significant differences compared to pre-operation.There were significant differences among two groups.At the final follow-up,the posterior muscle volume of the two groups at open side was decreased compared to preoperation,but there was no significant difference between the two groups.At the last follow-up,the posterior muscle volume at hinge side was not changed compared to preoperation in modified open-door laminoplasty group,while the volume of the posterior muscle at hinge side in the traditional control group decreased compared to preoperation.The difference was statistically significant.A total of eight patients with axial symptom were present after operation.There were two patients (6％,2/ 32) in the modified group and six patients (26％,6/23) in the control group.The difference of axial symptoms incidence was statistically significant between the two groups.Conclusion Modified open-door laminoplasty with preservation of the unilateral paraspinal muscle ligament complex is an effective technique in treating OPLL of the cervical spine.The technique not only guaranteed to have good recovery of neurological function and to maintain cervical curvature and range of motion,but also contributed to decrease the occurrence of postoperative axial symptoms.

The acute combination fractures of the atlas and axis in adults have a higher rate of neurological injury and early death compared with atlas or axial fractures alone. Currently, the diagnosis and treatment choices of acute combination fractures of the atlas and axis in adults are controversial because of the lack of standards for implementation. Non-operative treatments have a high incidence of bone nonunion and complications, while surgeries may easily lead to the injury of the vertebral artery, spinal cord and nerve root. At present, there are no evidence-based Chinese guidelines for the diagnosis and treatment of acute combination fractures of the atlas and axis in adults. To provide orthopedic surgeons with the most up-to-date and effective information in treating acute combination fractures of the atlas and axis in adults, the Spinal Trauma Group of Orthopedic Branch of Chinese Medical Doctor Association organized experts in the field of spinal trauma to develop the Evidence-based guideline for clinical diagnosis and treatment of acute combination fractures of the atlas and axis in adults ( version 2023) by referring to the "Management of acute combination fractures of the atlas and axis in adults" published by American Association of Neurological Surgeons (AANS)/Congress of Neurological Surgeons (CNS) in 2013 and the relevant Chinese and English literatures. Ten recommendations were made concerning the radiological diagnosis, stability judgment, treatment rules, treatment options and complications based on medical evidence, aiming to provide a reference for the diagnosis and treatment of acute combination fractures of the atlas and axis in adults.