BACKGROUNDIntervertebral disc degeneration is the main cause of low back pain. The purpose of this study was to explore potential methods for reversing the degeneration of lumbar intervertebral discs by transplantation of gene-modified nucleus pulposus cells into rabbit degenerative lumbar intervertebral discs after transfecting rabbit nucleus pulposus cells with adeno-associated virus 2 (AAV2)-mediated connective tissue growth factor (CTGF) and tissue inhibitor of metalloproteinases 1 (TIMP1) genes in vitro.

METHODSComputer tomography (CT)-guided percutaneous annulus fibrosus injury was performed to build degenerative lumbar intervertebral disc models in 60 New Zealand white rabbits. rAAV2-CTGF-IRES-TIMP1-transfected rabbit nucleus pulposus cells were transplanted into degenerative lumbar intervertebral discs (transplantation group), phosphate-buffered saline (PBS) was injected into degenerative lumbar intervertebral discs (degeneration control group) and normal lumbar intervertebral discs served as a blank control group. After 6, 10 and 14 weeks, the disc height index (DHI) and signal intensity in intervertebral discs were observed by X-ray and magnetic resonance imaging (MRI) analysis. The expression of CTGF and TIMP1 in nucleus pulposus tissue was determined by Western blotting analysis, the synthesis efficiency of proteoglycan was determined by a (35)S-sulfate incorporation assay, and the mRNA expression of type II collagen and proteoglycan was detected by RT-PCR.

RESULTSMRI confirmed that degenerative intervertebral discs appeared two weeks after percutaneous puncture. Transgenic nucleus pulposus cell transplantation could retard the rapid deterioration of the DHI. MRI indicated that degenerative intervertebral discs were relieved in the transplantation group compared with the degeneration control group. The expression of collagen II mRNA and proteoglycan mRNA was significantly higher in the transplantation group and the blank control group compared with the degeneration control group (P < 0.05).

CONCLUSIONSCT-guided percutaneous puncture can successfully build rabbit degenerative intervertebral disc models. Both CTGF and TIMP1-transfected cell transplantation helps to maintain disc height, and promotes the biosynthesis of type II collagen and proteoglycan in intervertebral discs, reversing the degeneration of intervertebral discs.

Animals ; Blotting, Western ; Cell Transplantation ; methods ; Cells, Cultured ; Collagen Type II ; genetics ; Connective Tissue Growth Factor ; genetics ; metabolism ; Dependovirus ; genetics ; Intervertebral Disc ; cytology ; diagnostic imaging ; metabolism ; Intervertebral Disc Degeneration ; diagnostic imaging ; metabolism ; therapy ; Magnetic Resonance Imaging ; Rabbits ; Radiography ; Reverse Transcriptase Polymerase Chain Reaction ; Tissue Inhibitor of Metalloproteinase-1 ; genetics ; metabolism

The research on the morphology features of the acetabulum bone joint surface area would be helpful to establishing the acetabulum 3D model for the purpose of the biomechanical analysis of hip joint, and therefore might have its important clinical significance. However, in former studies, the acetabulum was simply considered as a semi sphere. In this study, based on the acetabulum 3D-point data acquired by the 3D laser surface scanner and the reverse engineering technology together with the optimal fit algorithm, two kinds of best-fit model were achieved by a sphere surface and a rotating elliptical surface respectively approaching to the acetabulum bone joint surface. Both fitting errors were then compared and analyzed. The results showed that the fitting error of the rotating elliptical surface was significantly less than that of the sphere surface (P < 0.001). The average radius of fitting sphere was 24.37 +/- 2.22 mm and the average long axis of fitting rotating elliptical surface was 26.02 +/- 2.76 mm while its short axis was 24.17 +/- 2.16 mm. These findings would be helpful to our new recognition of the acetabulum since they were results of the first quantitative analyses for the acetabulum bone surface and also might serve as an important reference base in its further studies and application.
Acetabulum ; anatomy & histology ; Adult ; Algorithms ; Humans ; Imaging, Three-Dimensional ; Models, Anatomic

OBJECTIVETo explore the prevalence, distribution and clinical significance of high-intensity zone (HIZ) in anterior annulus fibrosus (AF) in comparison with HIZ in posterior AF of lumbar disc.

METHODSAccording to the diagnosis and location of HIZ, 610 lumbar magnetic resonance images with entire clinical materials were divided into control group (without HIZ), anterior AF group (HIZ), posterior AF group (HIZ) and anterior & posterior (AP) AF group (HIZ). The incidence of HIZ was summarized. The clinical data, such as male/female ratio, age, and body weight, prevalence of low back pain (LBP) and distribution of HIZ, were compared and analyzed between the groups.

RESULTSThree hundred and fifteen cases shown no HIZ (51.6%), 95 cases presented HIZ in anterior AF (15.6%, 119 discs), 159 cases presented HIZ in posterior AF (26.1%, 189 discs) and 41 cases presented HIZs in both anterior and posterior AF (6.7%, 96 discs). There was significant difference between the prevalence of HIZ in anterior AF and that in posterior AF (P < 0.01). The male/female ratio and body weight of each groups showed no difference (P > 0.05), and the age was proved to be statistically different between four groups (P < 0.01, control group < posterior AF group < AP AF group < anterior AF group). HIZs in anterior AF often occurred at L(1,2)-L(4,5), whereas, they usually developed at L(3,4)-L(5)S(1) in posterior AF. The incidence of LBP in control group, anterior AF group, posterior AF group, AP AF group were 40.0%, 52.6%, 55.4% and 65.8%, respectively. The LBP prevalence of control group was lower than that of other three groups (P < 0.05), and the prevalence of last three groups showed no difference (P > 0.05).

CONCLUSIONSCompared with HIZ in posterior AF, the HIZ in anterior AF of lumbar disc has a lower prevalence, often develops in elder patients and in upper motion segments. It also indicates an obvious relation to LBP as the former.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Intervertebral Disc ; pathology ; Intervertebral Disc Displacement ; diagnosis ; pathology ; Lumbar Vertebrae ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Young Adult

OBJECTIVESince 2011 EB-APS conference, we hypotheses that phase switching of inspiration-expiration is dominantly initiated by oscillatory information PaO2, PaCO2 and [H+] via fast peripheral chemical receptors. However, the evidence of the waveform of ABG is lack.

METHODSSix surgery patients with normal heart function and negative Allen test, had been placed the arterial catheterization directly connected to 3 x 1 000 mm pre-heparin plastic pipe for continuous collecting arterial blood. We counted the number of heart beat for the blood collecting time, and separated the blood pipe into the heart beat numbers' short pieces using haemostatic forceps, then put pipe into iced water at once fir analyzing PaO2, PaCO2, pH and SaO2 as soon as possible. We selected two breaths cycles of waveform from each patient for data calculations of magnitudes and time interval.

RESULTSThe heart beat numbers for filling blood into pipe were 16 ± 2, and all covered more than 2 breathing cycles. Each breathing cycle is cover 5 ± 0.6 heart beat. There were significant changes of PaO2, PaCO2, [H+] a and SaO2 (i.e. the highest high values compare to the next lowest values, P < 0.05). The time interval of changing PaO2, PaCO2, [H+]a and SaO2 magnitudes were 11.28 ± 1.13 mmHg, 1.77 ± 0.89 mmHg, 1.14 ± 0.35 nmol/L and 0.52% ± 0.44% respectively.

CONCLUSIONThis simple continuous beat-by-beat arterial blood sampling and ABG analyzing method is new and practicable. We obtain a clear evidence of periodic parameters ABG waveform, which following breathing cycle.

Arteries ; physiology ; Blood Gas Analysis ; Heart Rate ; Humans ; Monitoring, Physiologic ; methods ; Respiration

OBJECTIVETo construct adeno-associated virus (AAV) expression system for transforming growth factor beta3 (TGFbeta3 ) and detect its biological effect on proteoglycan synthesis of the earlier and later dedifferentiated rabbit lumbar disc nucleus pulpous (NP) cells, which was compared with that of adenovirus (AV) expression system for TGFbeta1.

METHODSTGFbeta3 gene was obtained using PCR. Its upstream contained restriction enzyme site Kpn I, and its downstream contained restriction enzyme site Sal I. Using the restriction enzyme sites of PCR product of TGFbeta3 and the corresponding multiple cloning site (MCS) in plasmid AAV, TGFbeta3 was subcloned into AAV. The recombinant plasmid AAV-TGFbeta3 was transfected into H293 cells with Lipofectamine 2000, and the expression of TGFbeta3 gene was detected using immunofluorescent analysis. After AAV-TGFbeta3 virus particle with infectious activity was packaged, TGFbeta3 expression in NP cells was detected by immunoblotting, and its biological effect on proteoglycan synthesis was detected by antonopulos method and compared with that of AV-TGFbeta1 in the earlier and later dedifferentiated NP cells.

RESULTSFor the earlier dedifferentiated NP cells, AAV-TGFbeta3 slowly and stably enhanced proteoglycan synthesis, but AV-TGFbeta1 rapidly and transiently enhanced its synthesis. For the later dedifferentiated NP cells, AAV-TGFbeta3 stably enhanced proteoglycan synthesis, but AV-TGFbeta1 inhibited its synthesis.

CONCLUSIONAAV expression system can mediate TGFbeta3 gene to be expressed stably, and AAV-TGFbeta3 can enhance proteoglycan synthesis of the earlier and later dedifferentiated NP cells.

Animals ; Cell Line ; DNA, Recombinant ; genetics ; Dependovirus ; genetics ; metabolism ; Female ; Humans ; Intervertebral Disc ; cytology ; Placenta ; cytology ; Plasmids ; genetics ; Polymerase Chain Reaction ; Pregnancy ; Proteoglycans ; biosynthesis ; Rabbits ; Transforming Growth Factor beta1 ; genetics ; Transforming Growth Factor beta3 ; genetics ; Viral Proteins ; biosynthesis

OBJECTIVETo study gene expression of collagen types IX and X in human lumbar intervertebral discs during aging and degeneration and to explore the role of collagen types IX and X in disc degeneration.

METHODSFetal, adult and pathologic specimens were subjected to in situ hybridization with cDNA probes to investigate mRNA-expressions of types IX and X collagen gene.

RESULTSIn fetal intervertebral discs, positive mRNA hybridization signals of type IX collagen were concentrated in the nucleus pulposus and the inner layer of anulus fibrosus. Interstitial matrix of the nucleus pulposus also showed positive type X collagen staining. Positive mRNA hybridization signals of types IX and X were not detected in the middle and outer layers of anulus fibrosus. In adult specimens, expression of type IX collagen mRNA was markedly decreased. No hybridization signals of type X collagen was observed. As for pathological specimens, there was no gene expression of type IX collagen. In severe degenerated discs from adults, there were focal positive expressions of type X collagen.

CONCLUSIONSObvious changes of collagen gene expression occur with aging. Expression of type IX collagen decreases in adult and pathological discs. Results of type X collagen expression suggest that type X collagen is expressed only in older adult and senile discs (i.e., when disc degeneration has already reached a terminal stage), indicating the terminal stage of degeneration.

Adolescent ; Adult ; Collagen Type IX ; metabolism ; Collagen Type X ; metabolism ; Female ; Gene Expression ; Humans ; Immunohistochemistry ; In Situ Hybridization ; Intervertebral Disc ; embryology ; metabolism ; Lumbar Vertebrae ; Male

BACKGROUNDLow back pain has emerged as a widespread disease often caused by intervertebral disc degeneration. This study aimed to establish an in vitro cell culture model of rhesus monkey lumbar intervertebral discs and to investigate the effect of combined connective tissue growth factor (CTGF) and tissue inhibitor of metalloprotease-1 (TIMP-1) expression mediated by adeno-associated virus (AAV) on collagen type II and proteoglycan levels. The purpose of these investigations was to explore potential methods for relieving the degeneration of lumbar intervertebral disc cells.

METHODSRhesus monkey lumbar intervertebral disc nucleus pulposus cells (NPCs) were isolated by enzyme digestion, cultured, and transduced with rAAV2-CTGF-IRES-TIMP-1, rAAV2-CTGF, or rAAV2-TIMP-1 at a multiplicity of infection (MOI) of 10(6). The expression of collagen type II and proteoglycan was measured using RT-PCR and Western blotting. The synthetic rate of proteoglycan was measured using (35)S incorporation.

RESULTSRhesus monkey lumbar intervertebral disc NPCs were transduced with rAAV2-CTGF-IRES-TIMP-1, rAAV2-CTGF, and rAAV2-TIMP-1 and the transduced genes were expressed and detected. Compared to the control, CTGF promoted the synthesis of collagen type II and proteoglycan. TIMP-1 showed an enhancing effect on the expression of proteoglycan but no effect on collagen type II. Expression of both genes in rhesus monkey lumbar intervertebral disc NPCs significantly enhances the synthesis of proteoglycan and collagen type II.

CONCLUSIONSSingle gene transduction of CTGF or TIMP-1 can enhanced synthesis of proteoglycan. CTGF expression can also enhance collagen type II protein synthesis. Combined transduction of both CTGF and TIMP1 can significantly promote the expression of proteoglycan and collagen type II to levels greater than transduction of a single gene alone. Our study provides a good basis for multi-gene therapy to treat lumbar intervertebral disc degeneration.

Animals ; Blotting, Western ; Cells, Cultured ; Collagen Type II ; biosynthesis ; Connective Tissue Growth Factor ; genetics ; metabolism ; Intervertebral Disc ; cytology ; Macaca mulatta ; Proteoglycans ; biosynthesis ; Reverse Transcriptase Polymerase Chain Reaction ; Tissue Inhibitor of Metalloproteinase-1 ; genetics ; metabolism ; Transduction, Genetic

OBJECTIVETo study the outcome of laminoforaminotomy with posterolateral discectomy for patients with lateral disc herniation at C(7)-T(1).

METHODSFrom August 2000 to August 2008, 12 patients with lateral disc herniation at C(7)-T(1) underwent posterolateral discectomy were analyzed retrospectively. Neurologic function were evaluated with the Motor Scoring System. Preoperative motor were compared with postoperative one. The unique clinical manifestation, imageology features and intraoperative findings were analyzed.

RESULTSAll these twelve patients were lateral type. All the patients showed hand intrinsic muscles atrophy and hand weakness. Nine patients had no paraesthesia. The average follow-up period was 26 months. Postoperative scores were significantly higher than preoperative ones.

CONCLUSIONSDisc herniation at C(7)-T(1) is predominantly lateral type and present C(8) nerve motor deficit (hand intrinsic muscles atrophy and hand weakness) and only minority has paraesthesia in C(8) nerve dermatome. Posterolateral cervical discectomy technique is safe and effective for patients with lateral disc herniation at C(7)-T(1).

Adult ; Cervical Vertebrae ; Female ; Follow-Up Studies ; Humans ; Intervertebral Disc Displacement ; diagnosis ; surgery ; Male ; Middle Aged ; Retrospective Studies ; Thoracic Vertebrae ; Treatment Outcome

OBJECTIVETo retrospectively analyze and evaluate the results of treatment for atlantoaxial instability or dislocation employing pedicle screws of atlas and axis.

METHODSThirty-one patients (23 male and 8 female) with atlantoaxial instability or dislocation were stabilized using pedicle screws of atlas and axis between May 2005 to January 2008. The patients ranged in age from 17 to 67 years (mean 43.5 years). Patients consisted of chronic odontoid fracture in 17, Os odontoideum in 8, fresh odontoid fracture in 4, transverse ligament rupture in 1, rheumatoid arthritis in 1. Clinical features included neck pain in 31; restricted neck movement in 28, varying degrees of spastic quadriparesis in 19. All patients underwent posterior C(1) to C(2) pedicle screw fixation. Operative time, intraoperative blood loss, complications were recorded, neurological and radiographic studies were carried.

RESULTSMean follow-up time was 13 months. Operative time averaged 2.5 h. Mean intraoperative blood loss was 300 ml. A patient had postoperative wound infection and was treated conservatively with antibiotics and local wound care. A patient developed pulmonary artery embolism and got well with anticoagulation. Satisfactory stability was achieved in all cases with no vascular and C(2) neuralgia. Average JOA score in 19 cases increased at final follow-up (P < 0.01). Solid fusion was achieved in 29 cases, fusion rate was 93.6%.

CONCLUSIONSStabilization of atlantoaxial complex via pedicle screws of atlas and axis has advantages of intraoperative restoration, easier placement of screw, solid fixation. It is a safe and effective treatment modality for posterior C(1-2) fusion.

Adolescent ; Adult ; Aged ; Atlanto-Axial Joint ; Bone Screws ; Female ; Follow-Up Studies ; Humans ; Joint Dislocations ; surgery ; Joint Instability ; surgery ; Male ; Middle Aged ; Retrospective Studies ; Spinal Fusion ; methods ; Treatment Outcome ; Young Adult

OBJECTIVETo investigate the neuroprotective effects of glycyrrhizin (Gly) as well as its effect on expression of high-mobility group box 1 (HMGB1) in rats after traumatic brain injury (TBI).

METHODSMale Sprague-Dawley rats were randomly divided into three groups: sham group, TBI group, and TBI+Gly group (n=36 per group). Rat TBI model was made by using the modified Feeney's method. In TBI+Gly group, Gly was administered intravenously at a dosage of 10 mg/kg 30 min after TBI. At 24 h after TBI, motor function and brain water content were evaluated. Meanwhile, HMGB1/HMGB1 receptors including toll-like receptor 4 (TLR4) and receptor for advanced glycation end products (RAGE)/nuclear factor-κB(NF-κB) signaling pathway and inflammatory cytokines in the injured brain tissues were detected using quantitative real-time polymerase chain reaction, western blot, electrophoretic mobility shift assay and enzyme-linked immunosorbent assay. Furthermore, HMGB1, RAGE and TLR4 immunohistochemistry and apoptosis were analyzed.

RESULTSBeam walking performance impairment and brain edema were significantly reduced in TBI+Gly group compared with TBI group; meanwhile, the over-expressions of HMGB1/HMGB1 receptors (TLR4 and RAGE)/NF-κB DNA-binding activity and inflammatory cytokines were inhibited. The percentages of HMGB1, RAGE and TLR4-positive cells and apoptotic cells were respectively 58.37% ± 5.06%, 54.15% ± 4.65%, 65.50% ± 4.83%, 52.02% ± 4.63% in TBI group and 39.99% ± 4.99%, 34.87% ± 5.02%, 43.33% ± 4.54%, 37.84% ± 5.16% in TBI+Gly group (all P<0.01 compared with TBI group).

CONCLUSIONGly can reduce secondary brain injury and improve outcomes in rat following TBI by down-regulation of HMGB1/HMGB1 receptors (TLR4 and RAGE)/NF-κB-mediated inflammatory responses in the injured rat brain.

Animals ; Brain Injuries ; drug therapy ; Glycyrrhizic Acid ; pharmacology ; therapeutic use ; HMGB1 Protein ; metabolism ; Male ; Neuroprotective Agents ; therapeutic use ; Rats ; Rats, Sprague-Dawley