Objective To investigate retrospectively the long-term results of discectomy for patients with lumbar intervertebral disc herniation. Methods From July 1988 to May 2003, 273 cases of 1040 patients with lumbar intervertebral disc herniation undergone surgical treatment in our hospital were followed up. All patients were divided three groups according the time of follow-up. The follow-up time was three years as middle follow-up group (Ⅰ), five years as longer follow-up group(Ⅱ) and ten years and more as sup-longer follow-up group (Ⅲ). Sixty-eight cases(24.91%) were in group Ⅰ, including 42 males and 26 females, with the average age of 43.7 years (14-63 years). The group Ⅱ included 141 cases (51.65%), 92 males and 49 females, with the average age of 46.1 years (18-76 years). As group ⅡⅢ, 64 cases (23.44%) were included 46 males and 18 females, with the average age of 43.5 years (20-63 years). The standards Scoring System of Chinese Spinal Association (CSA) and Japan Orthopaedic Association (JOA) were used for investigation. Results According to CSA system, the total good and excellent rate of surgical treatment for lumbar intervertebral disc herniation was 89.0%. The percentage of the satisfactory of the group Ⅰ, Ⅱ, Ⅲ were 92.6%, 91.5% and 79.7% respectively. There was significant difference between group Ⅰ and group Ⅱ, Ⅲ. The score of JOA were 24.75±5.08, 22.43±6.55, 21.64±7.18 postoperatively, with significant difference between group Ⅰ and group Ⅱ, Ⅲ. Conclusion The mid-term results of surgery for patients with lumbar iutervertebral disc herniation is good, and the good and excellent rate decreases gradually with the follow-up time. The results were similar to each other for evaluation between the standard of CSA and JOA.

0.05)between left AF(238,36.62%)and right AF(220,33.85%).More HIZs(446,68.62%)were located in inferior AF than that of middle or superior AF.The motion segments from L3、4 to L5S1 were the region that the HIZ occurred frequently and it could present in single segment or multi-segment.In anterior AF,HIZs often occurred at L2、3 and/or L3、4 discs.Whereas,they usually developed at L4、5 and/or L5S1 in posterior AF. Conclusion The incidence rate of HIZ in lumbar disc is higher.Posterior and inferior AF of discs and lower motive segments have more risk of HIZs.It could develop in single motive segment or multi-segments at one time.

Objective To study the biological effects of pSNAV2-hTGF?1 and pSNAV2-hTGF?3 on the reversion of rabbit disc degeneration. Methods Rabbit nucleus pulpous and annulus fibrosus cells were isolated and cultured. The fluorescence labled pSNAV2 were used to detect the transfect rates of rabbit disc cells at first. Then, the pSNAV2-hTGF?1 and pSNAV2-hTGF?3 were transfected into the degenerated rabbit disc cells respectively. The biological effects of hTGF?1 and hTGF?3 on degenerated rabbit disc cells were detected with Western-bloting and 35S detection to analyze and compare the matrix synthesis of the tranfected cells. Results pSNAV2 could transfect degenerated disc cells effectively in the early stages. Both the pSNAV2-hTGF?1 and pSNAV2-hTGF?3 could stimulate the synthesis of collagen Ⅱ and proteoglycan of the rabbit disc cells. For the early stage of degenerated disc cells, the synthesis of collagen Ⅱ and proteoglycan were greater transfected with pSNAV2-hTGF?1 than transfected with pSNAV2-hTGF?3. The pSNAV2-hTGF?1 could promote the degenerated rabbit annulus fibrosus cells to synthesize collagen Ⅰ and pSNAV2-hTGF?3 could promote the degenerated nucleus pulpous cells of later stage to synthesize the collagen Ⅱ. Conclusion Both pSNAV2-hTGF?1 and pSNAV2-hTGF?3 can promote the degenerated rabbit disc cells of early stage to synthesize the matrix. pSNAV2-hTGF?3 can efficently promote the seriously degenerated nucleus pulpous cells to synthesize the collagen Ⅱ.

Objective To deliver an exogenous therapeutic gene—hTGF ?1 to rabbit lumbar intervertebral disc in vivo by adenovirus vector and observe the expression of the exogenous therapeutic gene. Methods 20?l of 0 01?mol/L PBS, with or without adenovirus (6?10 6pfu) carrying TGF ?1 gene (Ad/CMV hTGF ?1) which were constructed by our laboratory, was injected directly into nucleus pulposus tissues of lumber discs of mature New Zealand white rabbits. Immunohistochemical staining for human transforming growth factor ?1 was performed on the rabbit disc tissues in different periods after operation. A half quantitation for immunohistochemical staining was performed by VIDAS analysis system. Results Discs injected with Ad/CMV hTGF ?1 exhibited extensive and intense positive immunohistochemical staining for transforming growth factor ?1 from 1 week to 12 weeks after operation, and positive pellets in nucleus pulposus cells in 4th day group, while the control groups(intact group and PBS group) showed negative or weakly positive immunohistochemical staining. The OPTDM was significantly increased in the discs injected with Ad/CMV hTGF?1 compared to the contact discs or the discs injected with only PBS( P

Objective To determine sensitive indices for defining degenerative cervical spinal canal stenosis(DCSCS) with a new radiograghic measurement method. Methods One hundred normal lateral radiographs of the cervical spine were divided into two groups according to ages. The following indices were measured or calculated: sagittal diameter of spinal canal(a), sagittal diameter of cervical body(b), sagittal diameter of edge or degenerative cervical body(c), cervical spinal canal ratio[(a/b),CSCR] and effective cervical spinal canal ratio [(a+b-c)/c,ECSCR]. The comparisons of these indices between two groups were performed to select the most sensitive index for clinical reference. Results There was no significant difference between the two age groups in terms of a, b and a/b. The comparison of c and (a+b-c)/c between two age groups showed significant difference at the level of C4,C5 and C6 (P

Objective To define the role of melatonin in the pathogenesis of chickens scoliosis following pinealectomy and constant light irradiation. Methods Ten white leghorn chickens in the control group were kept in light-dark (12h:12h) cycle, 500 lx in daytime and 0-5 lx in nighttime after birth. Pinealectomy was performed in 20 white leghorn chickens when 3-day-old and then kept in light-dark cycle as the control group. Constant light (500 lx) irradiation was used to reduce the secretion of melatonin in 20 chickens after their births. Radiologic examinations were performed on all chicken spines for scoliosis monthly. When the chickens were 3-month-old, their mid-day and mid-night serum samples were collected and analyzed with ELISA kit for melatonin. Results There was no scoliosis in the control group and constant light group when the chickens were 3-month-old. In the pinealectomy group, 4 chickens had obvious scoliosis in the first month when X-ray examination was taken. The curved deformity progressed and became serious when the chickens grew up. There were 7 chickens with severe curved deformity in the second month. When the chickens were 3-month-old, there were totally 11 chickens with scoliosis, Cobb' angle 11?-85?, average 30.63?. The level of melatonin in control group was low in daytime (10.6 pg/ml) and high in nighttime (110.4 pg/ml) alternately. The melatonin level was much lower, daytime 8.4 pg/ml and nighttime 6.9 pg/ml in pinealectomy group and 10.8 pg/ml in constant light group. There was no statistical significance in the serum melatonin between the pinealectomy group and constant light group. Both groups remained low level of serum melatonin. Conclusion Pinealectomy can reduce the secretion of melatonin and induce scoliosis in chickens. Although constant light could suppress the secretion of melatonin in chicken serum, it did not induce scoliosis. The pathogenesis of chickens scoliosis might not be mediated by low-level melatonin.

Objective To evaluate the reversion possibility of hVEGF165 and TGF?1 to intervertebral disc degeneration by gene method. Methods The hVEGF165cDNA obtained from plasmid pcDNA3(+)-hVEGF165 was subcloned into the packaging plasmid pSNAV of AAV by molecular clone ways. The recombinant plasmid pSNAV-hVEGF165 was identified by restriction enzymes analysis and sequencing analysis, and then transfered to the HEK293 cell and VEC by lipofectamine mediated gene transfer method. The protein hVEGF165 was detected by immunofluorescence for immunocytochemistry and explored the influence to the proliferation of vascular endothelial cell by MTT. Whereafter the AAV-hVEGF165 was packaged by Benyuan Zhengyang Company. AAV-hVEGF165 and AAV-TGF?1 were cotransfected into annulus fibrosus cell of intervertebral disc, then the expression of hVEGF165 and TGF?1, and the change of collagen Ⅰin annulus fibrosus cell were detected by Western blot. Results The recombinant pSNAV-hVEGF165 was completely constructed and confirmed by restriction enzymes analysis and sequencing analysis. The protein hVEGF165 was detected by immunofluorescence for immunocytochemistry in experimental group, and hVEGF165 could promote the proliferation of vascular endothelial cell. The bioactive AAV-hVEGF165 was successfully constructed. The expression of AAV-hVEGF165 and AAV-TGF?1 were manifested in degenerative annulus fibrosus cell by Western blot, and the expression of collagen Ⅰin annulus fibrosus cell cotransfected by AAV-hVEGF165 and AAV-TGF?1 was markedly more than that of the monogenic transfected cell. Conclusion hVEGF165 could cooperate with TGF?1 to promote the expression of collagen Ⅰ.

Objective\ To observe the expression of coreprotein gene in human lumbar discs in foetus and adult. Methods\ The coreprotein cDNA probe was prepared by RT-PCR. The mRNA level and distribution of coreprotein was observed in fetal and adult lumbar discs, using in situ hybridization method. Results\ 1) The highest expression lies in the lumbar nucleus pulposus of the fetal discs, less higher expression in the interior of the annulus fibrosus and the region between nucleus and annulus fibroses. 2) In adult disc tissue, few positive signals were shown. Low gene expression level of coreprotein in the subcultured disc cells was expressed. Conclusion\ The higher expression of coreprotein in fetal discs is chiefly located in the nucleus pulposus.

BACKGROUND: There are no reports about adult degenerative disc cell culture model currently. This establishment of disc cell culture is the cellular basis of intervertebral disc tissue engineering. OBJECTIVE: To establish cell culture models from human degenerative intervertebral disc, and to settle the cytology base of disc tissue engineering. DESIGN AND SETTING: The controlled observational experiment with regard to cytology was performed in Shandong Institute of Orthopaedics and Traumatology. MATERIALS: Five degenerative intervertebral disc patients were recruited from Department of Orthopaedics, Affiliated Hospital of Qingdao University Medical College, including one case in L2-3, three cases in L4-5 and one case in L5-S1. METHODS: Five samples were taken from the operational disectomy for lumbar disc herniation, and these samples were cultured using enzymatic digestion cell culture and tissue cell culture method differently. All the samples were cultured with HAMF12 medium with the addition of 10% and 20% fetal bovine serum. The morphology of the cultured cell was observed with Giernsa staining and transmission electron microscopy respectively. MAIN OUTCOME MEASURES: ①Cellular growth; ②Morphology of the cultured cells;③Ultrastructure Of the cultured cells. RESULTS: By means of enzymatic digestion cell culture and tissue cell culture method, numerous degenerative lumbar disc cells were obtained and successfully subcultured. The cell secretions around the cultured cells may influence the growth of these cells. Destroying the secretions, the cultured cells could be highly proliferated. The degenerative intervertebral disc cells were proliferated vigorously in HAMF12 medium with 20% fetal bovine serum compared with that with 10% fetal bovine serum. The notochordal cell was observed in all the specimens. CONCLUSION: The adult degenerative disc can be used to obtain the required cell for tissue engineering.

Objective 1) To verify the the potential of the adenoassociated viral vector as a strategy for intradiscal gene transfer in degenerative rabbit intervertebral discs. 2) To investigate the gene transduction efficacy and to quantify the biologic effects on the matrix synthesis after single gene transfer and combined gene transfer. Methods Rabbit models of disc degeneration were established by injecting the N-terminal 30×103 fibronectin fragment (Fn-f), 4 weeks later, saline with or without virus was injected directly into 144 lumbar discs of 36 skeletally mature New Zealand white rabbits. Group A (n =8) received the rAAV2-hTGFβ1; Group B (n=6) received rAAV2-hTGFβ3;Group C (n=6) recived rAAV2-hTGFβ1 and rAAV2-hTGFβ3; Group D (n=8) recived rAAV2-EGFP as the experimental control. Group E (n=8) recived PBS as the blank control. Two rabbits of the group A and group E were sacrefied 1 week after injection, immunohistochemical staining for hTGF-β1 was performed on the slices of nucleus pulposus (NP) tissues. On 4,8 and 12 weeks after gene transferring, NP tissues were cultured or decomposed to quantify the biochemical changes of the matrix using 35S-sulfate incorporation assay and western blot detection. The expression of EGFP was observed 12 weeks after injection. Results Discs in group A exhibited extensive and intense positive immunostaining for hTGF-β1 than the control discs in group E 1 week after gene transferring. The nucleus pulposus tissues in group A, B and C exhibited a 1.28-2.06 fold increase in proteoglycan synthesis and a 1.25-1.73 fold increase in collagen type Ⅱ production over those in group E (P＜0.05 or P＜0.01).Combination of two gene transfer in group C makes a significantly increased level of proteoglycan (1.195-1.290 fold)and collagen type Ⅱ (1.152-1.219 fold) than single gene transfer in group A and B(P＜0.05 or P＜0.01).No statistic differences shows between A group and B group. The difference of the matrix synthesis between group D and group E was also not statistically significant (P＞0.05). Extensive and intensive green fluorescence was observed on the slice of nucleus pulposus tissues received rAAV2-EGFP 12 weeks after gene delivery. The expression of EGFP kept for more than 12 weeks. Conclusion Findings showed that the disc tissue injected with rAAV2 mediated genes highly expressed the therapeutic proteins from 1 week to more than 12 weeks after delivery. It is suggested that adenoassociated virus be an valid vector for the transfer of the exogenous genes in the degenerative disc. The therapeutic factors hTGF-β1 and hTGF-β3 could efficiently increase the synthesis of proteoglycan and collagen type Ⅱ in the degenerative NP cells and combined transfer of two genes was more effective than single gene transfer. The two factors have an positive synergistic effects.