BACKGROUND:As the main function cel of intervertebral disc, nucleus pulposus cel s are the focus of studying the degenerative mechanism;thereby, it is crucial to maintaining the physiological function of nucleus pulposus cel s in vitro and the stability of the cel phenotype.
OBJECTIVE:To study the excel ence of differential velocity adherent procedure in primary culture of nucleus pulposus cel s of rat intervertebral disc through comparison.
METHODS:Twenty male Wistar rats aged 4 weeks were enrol ed, and then nucleus pulposus cel s of intervertebral disc were isolated and cultured in vitro;cel passage culture was performed in different groups when the primary cel s were merged to 90%. Differential velocity adherent group cel s adhered for 30 minutes, and non-adherent cel s were aspirated and transferred to new culture dish after readjusting the concentration;the controls received no intervention. Passages 1 and 2 cel s in the differential velocity adherent group were isolated and purified by the same procedure. The morphology of three generations of cel s in the two groups was compared, the purity of the identification was detected by immunohistochemistry, the cel viability was detected by cel counting kit-8 and the cel growth curve was drawn.
RESULTS AND CONCLUSION:Inverted phase contrast microscope and hematoxylin-eosin staining revealed that the cel homogeneity of the differential velocity adherent group was significantly higher than that of the control group, and there were more kinds of fibroblast-like cel s in nucleus pulposus cel s in the control group. Identification and purity analysis of col agen type II showed that the cytoplasm of two groups were both stained brown, indicating that they were the nucleus pulposus chrondrocytes. The positive rate of differential velocity adherent group was significantly higher than that of the control group (P<0.05). The cel growth curve of cel counting kit-8 showed cel s in the two groups al passed by the latency phase within 2 days, then to the logarithmic phase of 3 days, and entered the lag phase, while the growth rate of the control group was more rapid during the latency and the early logarithmic phases. These findings suggest that differential velocity adherence method is a practical and effective procedure for the isolation and purification of primary cultured rat nucleus pulposus cel s. Through the primary culture, twice differential velocity adherence procedure, the passage 3 rat nucleus pulposus cel s are metabolic exuberant, consistent with the phenotype, the cel purity is higher, and the logarithmic growth phase can be used as the optimal time for studying the mechanism of intervertebral disc cel s.

BACKGROUND:The narrowing of intervertebral space induced by the intervertebral disc degeneration is mainly characterized by the expression of proteoglycan in nucleus pulposus cells and the reduction of col agen type II. OBJECTIVE:To quantitatively observe col agen type II protein in adult normal and degenerative intervertebral disc nucleus pulposus cells by immunofluorescence staining and safranin O staining.
METHODS:The nucleus pulposus specimens were col ected from adult scoliosis patients and patients with intervertebral disc protrusion, who were al volunteers. After culture, 26 cells in each patient were measured. There were 78 cells in both normal group and degeneration group. The normal and degenerative intervertebral disc nucleus pulposus cells were subjected to safranin“O”staining, and gray values were determined;intracellular col agen type II was detected by immunofluorescence staining.
RESULTS AND CONCLUSION:Immunofluorescence staining revealed that, degenerative intervertebral disc nucleus pulposus cells were only mildly stained, with the fuzzy staining, the shape was round, spindle, fusiform and irregular. There were a very smal amount of fluorescent particles within cells. The expression of col agen type II was decreased significantly compared with normal cells (P<0.05). Safranin O staining showed that, degenerative nucleus pulposus cells began to swel , the nuclei swel ed and were stained slightly, cellprocesses were prolonged, cytoplasmic dyeing was uneven accompanying with vacuole, celldisruption, scattered and chaotic distribution were visible, patches of necrosis were observed. The image gray value showed no significant difference compared with normal nucleus pulposus cells (P>0.05). The degenerative intervertebral disc nucleus pulposus cells have a smal quantity and partial y become apoptotic, the content of col egen type II protein is decreased significantly compared with normal nucleus pulposus cells.

Objective:To evaluate the feasibility of transforaminal endoscopic nerve root decompression for degenerative lumbar spinal stenosis (DLSS).Methods: From July 2011 to April 2016,96 cases of single segment DLSS were involved.All the patients had unilateral lower extremity neurological symptoms,signs,neurogenic intermittent claudication of less than 500 m.Imaging examinations (CT or MRI) or diagnostic nerve root block confirmed single segment degeneration.The mean age was (71.6±5.4) years,male: 55 cases,female: 41 cases.Their intraoperative blood loss,operation time,complications,ambulation time and discharge time were recorded.Leg pain VAS,ODI were used to evaluate the pain and lumbar function of the patients.The clinical efficacy was evaluated by Nakai evaluation.Results: All the patients were performed endoscopic decompression of the lateral recess and nerve root by removing the ventral part of the superior facet joint,the ligamentum flavum and the intervertebral disc.The decompression range was from the inferior edge of the upper pedicle to the superior edge of the lower pedicle.The nerve root was detected to have no compression and the pulse of nerve root returned to normal.The patient got ambulant on the operation day and discharged if he had no discomfort symptom.In the study,68 cases got follow up.The mean follow-up time was 12.1 months (6-63 months).The VAS at dif-ferent follow-up time points was improved relative to the baseline,and the difference was statistically significant (F=491.60,P<0.001).The ODI at different follow-up time points was improved relative to the baseline,and the difference was statistically significant (F=189.91,P<0.001).The excellent and good rates of Nakai evaluation were 79.4％ (excellent in 42 cases,good in 12 cases,fair in 10 cases and poor in 4 cases).The mean intraoperative blood loss was (49.29±11.86) mL.The mean operation time was (92.46±21.34) min.The mean ambulation time was 1.8 h.The mean discharge time was 2.3 days.Postoperative epidural hematoma was found in 1 case.Foot drop was found in 1 case.Second stage open surgery was performed in 6 cases.Conclusion: We can apply transforaminal endoscopic decompression for the patients of lumbar spinal stenosis who have unilateral nerve root irritation.Patients with transforaminal endoscopic decompression can get less surgical trauma,quick recovery and obtain good short-term outcome.

Scoliosis is a common deformity in neuromuscular disease, which usually has the characteristics of early onset age, severe degree of deformity, and rapid progression.Neuromuscular scoliosis often leads to serious damages to the quality of life, and results in the loss of walking, standing and sitting, and cardiopulmonary insufficiency.Surgical treatment can improve the quality of life for children with neuromuscular scoliosis, but surgical treatment is still challenging due to the complex surgery and many complicated diseases.The complications are much higher than those of idiopathic scoliosis.A multidisciplinary team is necessary in the surgical treatment of neuromuscular scoliosis to promptly and effectively reduce the complications.