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.

Objective To investigate the possible role of leukocytes, a important cellular component of the blood, in the induction of microcirculatory disturbances in noise-exposed cochleae. Methods Guinea pigs were exposed to a 4 kHz narrow band noise at 115 dB SPL for 4 hours, and the exposed cochleae were collected at 2 or 4 days after the noise exposure. Several techniques (cell viability assay, fluorescent labeling for blood plasma and for hair cell nuclei) were used to examine morphological changes in microvessels in the spiral lamina and in the stria vascularis as well as the hair cell nuclei. Results In noise-exposed cochleae, aggregatad leukocytes were found in the vessels of the spiral lamina. In the up-stream of the vessels with aggregated leukocytes, red blood cells were tightly packed, causing an increase in vessel diameters. Most leukocytes were located within the lumen of vessels, but some of them infiltrated the spiral lamina, usually alongside the vessels. Cell viability, assessed by trypan blue staining, showed that these plugged leukocytes lost their viability. In contrast, no leukocytes aggregated in the vessels of the stria vascularis, nor did leukocytes lose their viability. Conclusion In the cochleae exposed to intense noise, leukocytes were trapped in the vessels of the spiral lamina, leading to disturbance of blood flow. The involvement of leukocytes in local damaging processes and the fate of leukocytes need further studies.

Objective To quantitatively analyze the occurrence of apoptotic and necrotic outer hair cells (OHCs) and to evaluate the changes of these parameters with time after noise exposure. Methods Chinchillas were exposed to a narrow band noise at either 104 or 108 dB SPL for 1 hour. The animals were executed and dissected at either 1, 4 and 30 days after the noise exposure and the cochleas were collected for detection of OHC death mode. The apoptotic and necrotic OHCs were distinguished by examining the OHC nuclear morphology and confirmed by staining for caspase 3 activity or TUNEL assay. ABR thresholds for click stimuli were used to monitor changes in auditory function. Results The number of apoptotic cells were significantly greater than those of necrotic cells shortly after the noise exposure at day 1 for the 104 dB group, day1 and day4 for 108 dB group ( P =0 01, P =0 03, P =0 01) and the difference between the number of apoptotic cells and necrotic cells became statistically insignificant at day 4 and day 30 for the 104 dB group and day 30 for 108 dB group ( P =0 67, P =0 29, P =0 52). By day 30, apoptotic and necrotic pathologies continued, although in small quantity in both 104 dB group and 108 dB group. Conclusions The results of the study indicated that the early expansion of cochlear lesion is contributed primarily by apoptosis, whereas the later stage of lesion expansion is likely contributed equally by apoptosis and necrosis. The death of OHCs not only takes place during a noise exposure, but also continues for at least 30 days after noise exposure

Objective To examine the role of mitochondrial energy-conversion efficiency in regulating the initiation and execution of the apoptotic process in outer hair cells(OHCs)of cochlea following exposure to intense noise.Methods Seventeen adult chinchillas were used in present study.The animals were randomly assigned to one of the two groups,12 animals were exposed to noise and the remaining 5 animals without exposure to noise served as normal control.For all the animals,3-nitropropionic acid solution(3-NP,50mmol/L),an irreversible inhibitor of succinate dehydrogenase(SDH),was dropped onto the round window of the cochlea of the right ear using a micro-syringe to inhibit the mitochondrial energy production,to serve as the test ear.Artificial perilymph solution(AP)was dropped onto the round window of the cochlea of the left ear to serve as the control ear.16h after application of 3-NP and AP,the animals were exposed to 75 pairs of impulse noise at 155 dB pSPL.The cochleae were harvested 2h after the noise exposure.The cochlear basilar membrane was stained with propidium iodide(PI),a DNA intercalating fluorescent probe used to visualize the morphologic viability of hair cell nuclei.All the specimens were examined with a fluorescence microscope.Results In the 3-NP-treated cochlea,medium degree of nuclear condensation of OHCs appeared to be the dominant nuclear pathology,and only a few OHCs showed nuclear fragmentation in the damaged area of the cochlea.In contrast,the AP-treated control ear exhibited a large quantity of nuclear fragments,and a small quantity of medium degree of nuclear condensation.Conclusion The present study indicates that mitochondrial energy-conversion function plays an important role in regulating the apoptotic process.Disruption of the mitochondrial energy can not deter the apoptotic process from initiation,but can slow down the pace of apoptotic progression in outer hair cells of the cochlea following exposure to intense noise.

Objective Apoptosis and necrosis are two forms of hair cell death in cochlea following exposure to intense impulse noise.The present study was designed to examine the activity of caspase-3,caspase-8 and caspase-9 in the basilar membrane of cochlea,and investigate the cellular signal pathway associated with noise-induced hair cell death in cochlea.Methods Twenty-seven adult chinchillas were used in present study.The animals were randomly assigned into test group(no.15) and control group(no.12).The animals in test group were exposed to noise for the detection of the activity of caspase-3,caspase-8 and caspase-9,respectively,each with 5 animals.Animals in control group were not exposed to noise.Chinchillas were exposed to impulse noise at 155 dB peak sound pressure level(pSPL) for 75 times.Two hours after noise exposure,both cochleae were perfused with approximately 30?l of freshly prepared solution containing one of the three caspases(caspase-3,-8 and-9) respectively using a micro-syringe.One hour after perfusion,the animals were sacrificed and the cochleae were harvested.The Corti organs were double stained with propidium iodide(PI),and a DNA intercalating fluorescent probe was used to visualize the morphologic viability of hair cell nuclei.All the specimens were observed with a fluorescence microscope.Results The normal cochlea did not exhibit green fluorescence for any of the three caspases in Corti organ.Strong caspase-3,-8 and-9 green fluorescence appeared in the outer hair cells with condensed or fragmented nuclei,a sign of apoptosis,whereas no fluorescence was observed for any of the three caspases in the swollen outer hair cell nuclei,a sign of necrosis,in the cochleae exposed to intense impulse noise.Conclusion It is indicated that caspase-9 is involved in the intrinsic and caspase-8 involved in the extrinsic apoptotic pathway which occur simultaneously in apoptotic outer hair cells.No caspase activation occurs in necrotic outer hair cell in the cochlea following exposure to intense impulse noise.

Objective To compare the prevalence of apoptosis and necrosis, and investigate the primary death pathway of outer hair cells of rat cochlea following styrene exposure. Methods Fourteen adult Long Evans rats were used in the present study. The animals were randomly assigned into test group (n=8) and control group (n=6). Animals in test group were exposed to styrene by gavage at 400 mg/kg (2g styrene was mixed with 1ml olive oil). Treatment was performed once a day, 5 days per week for 3 weeks. Animals in control group were fed by gavage the same volume of olive oil on an identical time schedule used for the test group. The auditory brainstem response (ABR) thresholds of both ears elicited with clicks were measured before and at the end of the 3-week styrene or olive oil treatment. After hearing was re-assessed, animals were sacrificed and cochleae were quickly removed from the skull. Following fixation, whole specimens comprising the basilar membrane with Corti's organ were separated from the modiolus. Apoptotic, necrotic and missing outer hair cells (OHCs) were distinguished by combined assays of nuclear staining with propidium iodide (PI), TUNEL assay and filamentous actin(F-actin)staining with FITC-phalloidin. Each Corti's organ was thoroughly examined by fluorescence microscopy. The numbers of damaged OHCs (apoptotic, necrotic and missing OHCs) were documented. Results Neither threshold shift of ABR nor sign of hair cell (HC) damage was found in the cochlea of control animals. The animals of test group showed both physiological and pathological changes in the cochleae following the 3-week styrene treatment. ABR testing revealed an average of 15 dB of threshold shifts. F-actin staining exhibited the maximal level of OHCs damage in the middle portion of Corti's organ. The major damage occurred in the third row of OHCs, followed by the second and first rows of OHCs. Three types of morphological changes in damaged OHC nuclei were revealed by PI labeling: nuclear condensation, nuclear swelling and nuclear missing. Strong TUNEL green fluorescence appeared in the OHCs with condensed nuclei. Quantitative analysis showed that the average number of apoptotic OHCs was approximately three times greater than the number of necrotic OHCs (P=0.01). Conclusion It is indicated that apoptosis is the primary death pathway of OHCs leading to generation of the cochlear lesion following styrene exposure.

Objective To make a quantitative analysis of damaging hair cells,and investigate the progression pattern of cochlear pathology and the primary death pathway of hair cells in cochleae of aging Fischer 344 rats.Methods Thirty-two Fischer 344 rats were used in this experiment.The animals were assigned to two groups:rats in young group (n=13) were 3-4 months old,and in aging group (n=19) were 20-27 months old.Rats in aging group were further divided into two subgroups based on the animals' age:20-23-month subgroup (n=13) and 24-27-month subgroup (n=7).The auditory brainstem response (ABR) thresholds of both ears elicited with tone bursts at 5,10,20 and 40 kHz were measured in both young,and aging Fischer 344 rats.Upon completion of the auditory test,animals were decapitated,and both left and right bullae were exposed.Following fixation,whole specimens comprising the basilar membrane with Corti organ were separated from the modiolus.Propidium iodide (PI),a popular DNA intercalating fluorescent probe was used to trace the morphological changes in hair cell nuclei,and a terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay was used to detect nuclear DNA fragmentation in the aging cochleae.Filamentous actin (F-actin),an important structural protein of outer hair cells (OHCs),was stained with FITC-labeled phalloidin to illustrate the morphologic viability of cuticular plate and the stereocilia for affirming the missed OHCs.Each Corti organ was thoroughly inspected from the apical to the basal turns of the cochlea under a fluorescence microscope.The numbers of damaging OHCs including missed nuclei,condensed nuclei and swollen nuclei were documented,and a cochleogram was drawn.Results There were significant thresholds at all tested frequencies between the young and aging rats (P

Objective To investigate the clinical outcomes of trans-facet joints approach to treat thoracic degenerative disease with anterior compression.Methods From January 2003 to December 2009,22 patients with thoracic myelopathy caused by anterior compression were studied retrospectively.The patients included 16 males and 6 females,aged from 36 to 72 years(average 54.2 years).There were thoracic ossification of posterior longitudinal ligament(OPLL)in 11 cases,thoracic disc protrusion with ossification in 8 cases,thoracic vertebra posterior osteophytes in 2 cases,ankylosing spondylitis with thoracic pseudoarthrosis in 1 cases.Preoperative Japanese Orthopaedic Association(JOA)score was 5.2(range,2-9).The characteristic of thoracic degeneration was analyzed by CT and MRI examination.Posterior decompressive laminectomies were performed by the technique of "cap uncovering".The facet joints were removed bilaterally.Anterior ossified compressions were cut via posterior-lateral approach,and then intervertebral bone graft and bilateral pedicle screws were implanted.Results All patients were followed up for 8 to 38 months.According to the revised Epstein standard,there were excellent in 7 patients,good in 9,fair in 4,and poor in 2.The total effective rate was 90.9%(20/22).The excellent and good rate was 72.7%(16/22).The mean postoperative JOA score was 8.7(range,2-11).Surgical complications included dural laceration in 1 patient,pleura injury in 1 patient,epidural hematoma in 2 patients.There were no cases of spinal instability or deep infection.Conclusion The anterior compression can be solved completely via trans-facet joints approach in thoracic degenerative disease patients.

Objective To investigate the expression of DR5 and DcR2 protein and mRNA in lumbar intervertebral disc (LID) of patients with prolapse of LID and normal controls.Methods The expression and distribution of DR5 and DcR2 protein were immunostained in prolapsed LIDs of 60 patients and 22 normal LIDs of 8 normal controls. In parallel, mRNA of DR5 and DcR2 was quantified by real time fluorescent reverse transcriptase-polymerase chain reaction (RT-PCR) in prolapsed LIDs of 30 patients and 9 normal LIDs of 3 normal controls.Results The positive rates of DR5 protein in prolapsed IVDs and normal IVDs were 41.60% and 26.09% respectively. There were a significant difference between the two groups ( P=0.001). Further similar evidences were obtained by quantification of DR5 mRNA ( P=0.025). There was no difference in the expression of DcR2 protein and mRNA between the two groups.Conclusion The apoptosis pathway induced by DR5/tumor necrotic factor-related apoptosis-inducing ligand (TRAIL) maybe exists in LID tissues.

BACKGROUND:Inhibiting the apoptosis of intervertebral disc cel s can postpone the degenerative process of intervertebral disc. Survivin has a strong function of regulating cel proliferation and anti-apoptosis.
OBJECTIVE:To construct and identify the lentiviral vector encoding survivin gene of human.
METHODS:The survivin gene of human (BIRC5) was synthesized through the gene synthesis technology, amplified by PCR and analyzed by electrophoresis. The target gene was cloned into lentiviral expression plasmid to obtain the recombinant lentiviral vector Lenti-BIRC5. After transformation into competent E. coli cel s, the candidate clones were identified by PCR firstly. The positive clones were identified by gene sequencing. The lentivirus plasmid containing target gene was transfected into 293T cel s, and the expression of recombinant lentiviral vector Flag-Survivin fusion protein was detected through western blot analysis.
RESULTS AND CONCLUSION:The PCR results of electrophoresis and DNA sequencing showed that lentiviral vector containing human survivin gene was constructed successful y. Western blot analysis results showed that the target gene was transfected successful y and over-expressed in cultured cel s. The lentiviral expression vector of human survivin gene Lenti-BIRC5 was constructed successful y, which lays a foundation for the study addressing the anti-apoptotic effects of survivin on human nucleus pulposus cel s.