Multiplex analysis of serum hormone and cytokine in patients with cervical cOPLL: towards understanding the potential pathogenic mechanisms.

Cervical ossification of the posterior longitudinal ligament (cOPLL) is one of the major causes of myelopathy. However, the mechanism underlying remains elusive. In the present study, using MILLIPLEX magnetic bead panel, we investigated four serum hormones and six serum cytokines in cOPLL patients and healthy subjects. The results showed that tumor necrosis factore-α (TNF-α) were significantly increased, and DDK-1 was significantly decreased in the serum from male and female cOPLL patients compared with those from healthy controls, respectively. Osteopontin (OPN) and fibroblast growth factor-23 (FGF-23) were significantly increased in male cOPLL patients compared with that in healthy male controls. Further analysis showed that FGF-23 and OPN significantly increased, dickkopf-1 (DKK-1) decreased in the extensive cOPLL group. In addition, a significant positive correlation between the OPN and FGF-23 was observed in male cOPLL patients. The results are useful for understanding the mechanism underlying cOPLL.

CT evaluation of upper thoracic spine for surgical application of transarticular screw placement.

INTRODUCTION: Although pedicle screw fixation has been increasingly used in the upper thoracic spine in recent years, controversies exist about the safety and complications such as nerve or vascular intrusion associated with the technique. In this study, an alternative method of transarticular screw fixation was validated. MATERIALS AND METHODS: Morphometric analysis was performed on computed tomography (CT) scans of the upper thoracic zygapophysial joints of C7, T1, T2 and T3 in 20 male and 20 female patients in the axial and sagittal planes. The degree of screw angulation was recorded in the sagittal and axial planes and the screw length was measured at the spinal level from C7 to T3. RESULTS: The smallest medial-lateral diameter and anterior-posterior diameter of IAP was found at T3 in the female patients and C7 in the male patients. The screw trajectory length ranged from 14.9 to 20.5 mm in all patients. All the above measurements were significantly different between male and female patients at all levels (P < 0.05). The mean value of screw trajectory angle was 19.3°-20.1° in the axial plane and 44.3°-45.7° in the sagittal plane. There was no statistically significant difference (P > 0.05) between male and female patients in the axial and sagittal angles. CONCLUSION: The morphometric data of C7-T3 zygapophysial joints indicate the suitable screw diameter and screw length for this technique. Transarticular screw fixation proved to be a potentially safe alternative to pedicle screw fixation in this region.

GPBAR1 is associated with asynchronous bone metastasis and poor prognosis of hepatocellular carcinoma.

Background: Hepatocellular carcinoma (HCC) is the second leading cause of cancer-related death in China. Asynchronous metastasis is the main reason for HCC recurrence, but the current assessment of HCC metastasis and prognosis is far from clinically satisfactory. Materials: In our study, we investigated the expression of G-protein-coupled bile acid receptor (GPBAR1) in HCC tissues and tumor-adjacent tissues by qRT-PCR and immunohistochemistry. The associations between GPBAR1 expression, clinicopathological factors, and asynchronous metastases were assessed by the Chi-square test. The overall survival curves of different variables were plotted with the Kaplan-Meier method, and the statistical significance between different subgroups was analyzed with the log-rank test. The independent prognostic factors were identified by the Cox regression hazard model. Results: GPBAR1 was more highly expressed in HCC tissues than in tumor-adjacent tissues. GPBAR1 expression in HCC was significantly higher than that in liver cirrhosis, followed by normal liver tissues. GPBAR1 was significantly associated with poor prognosis in HCC and can be regarded as an independent prognostic biomarker. Interestingly, GPBAR1 expression in HCC was significantly correlated with asynchronous metastasis to the bone but not to the liver or lung. Conclusions: GPBAR1 was found to be an independent, unfavorable prognostic factor of HCC, as well as an indicator of asynchronous bone metastasis but not liver or lung metastases. Our results could provide a new aspect for HCC metastasis studies and help identify high-risk HCC patients, which helps ameliorate the prognostic assessment of HCC.

Simultaneous ossification of the posterior longitudinal ligament and ossification of the ligamentum flavum causing upper thoracic myelopathy in DISH: case report and literature review.

A rare case of a 44-year-old Chinese male with diffuse idiopathic skeletal hyperostosis (DISH) and simultaneous ossification of the posterior longitudinal ligament (OPLL) and ossification of the ligamentum flavum (OLF) at T1-2 causing thoracic myelopathy is reported herein. Posterior decompression without extirpating the OPLL was performed at T1-2. Postoperatively, symptoms were greatly improved, with remaining hyperreflexia and Grade 4/5 muscle strength in the lower extremities. The Japanese Orthopedic Association score improved from 5 preoperatively to 9 at final follow-up. The presence of a cyst due to leakage of cerebrospinal fluid was confirmed by MRI at day 27, but it resolved after conservative management. The clinical manifestation of DISH, the relationship among DISH, OPLL, and OLF, and management of thoracic myelopathy due to OPLL and OLF were reviewed.

Intra-discal vancomycin-loaded PLGA microsphere injection for MRSA discitis: an experimental study.

OBJECTIVE: To prepare the vancomycin hydrochloride (VA)-loaded poly lactic acid-glycolic acid (PLGA) copolymer microsphere by the multiple emulsion method and evaluate its therapeutic effects on infective discitis. METHODS: Firstly, the particle diameter distribution, shape, encapsulation efficiency, drug-loaded dosage and release curve of VA-PLGA microspheres were evaluated in vitro. Rabbits with methicillin-resistant Staphylococcus aureus infective discitis were treated with VA-PLGA intra-discal injection. Meanwhile, VA intravenous injection, blank PLGA microspheres intra-discal injection served as controls. Thirty days later, therapeutic effects were evaluated through X-ray radiophotography, histopathological and bacteriological examination. RESULTS: Mean particle diameter was between 61.57 ± 4.37 and 67.45 ± 8.13 µm, and mean encapsulation efficiency was between 60.20 ± 1.61 and 75.27 ± 1.60 %m/m. In vitro release experiment showed that the release time was over 30 days. The result of in vivo experiment showed that inflammatory reaction in the VA-PLGA intra-discal injection group was milder than the intravenous injection group (P < 0.05), also with less inflammation. The bacterial count was also significantly lower (1.02 × 10(3) ± 1.22 × 10(3) CFU/g) than the intravenous injection group (7.51 × 10(4) ± 7.16 × 10(4) CFU/g) (P < 0.05). Besides these data, the amount used in VA-PLGA intra-discal injection group is about 20 mg, and that used in the intravenous injection group is about 2.4 g. So, we just use 1/120 of VA i.v. to obtain the better results with our microparticles. CONCLUSION: Intra-discal injection with VA-PLGA sustained-release microspheres can use much less dosage, and effectively control and reduce infective discitis, and the therapeutic effect is superior to that of intravenous injection. A need for the clinical trials will be carried out in the near future.

Bilateral C1 laminar hooks combined with C2 pedicle screws fixation for treatment of C1-C2 instability not suitable for placement of transarticular screws.

The study design described here is a posterior C1-C2 fusion technique composed of bilateral C1 hooks and C2 pedicle screws. In addition, the clinical results of using this method on 13 patients with C1-C2 instability are reported. The objectives are to introduce a new technique for posterior C1-C2 fusion and to evaluate the clinical outcome of using it to treat C1-C2 instability. From October 2006 to August 2008, 13 patients (9 men and 4 women) with C1-C2 instability were included in this study: 3 had acute odontoid fractures, 4 had obsolete odontoid fractures, 4 had os odontoideum and 2 had traumatic rupture of the transverse ligament. All patients underwent posterior atlantoaxial fixation with bilateral C1 hooks and C2 pedicle screws. The mean follow-up duration was 25 months (range 13-30 months). Each patient underwent a complete cervical radiograph series, including anterior-posterior, lateral, and flexion-extension views, and a computed tomographic scan. The clinical course was evaluated according to the Frankel grading system. No clinically manifested injury of the nerve structures or the vertebral artery was observed in any of these cases. Five patients with neurological symptoms showed significant improvement in neurological function postoperatively. Bony fusion and construction stability were observed in all 13 patients (100%) on their follow-up radiographs, and no instrument failure was observed. Bilateral C1 hooks combined with C2 pedicle screws can be used as an alternative treatment method for C1-C2 dislocation, especially in cases not suitable for the use of transarticular screws. The clinical follow-up shows that this technique is a safe and effective method of treatment.

NgR expression in macrophages promotes nerve regeneration after spinal cord injury in rats.

OBJECTIVE: This study aimed to investigate the expression of Nogo-66 receptor (NgR) in macrophages after SCI and clarify its role in neuron regeneration. METHODS: Macrophages harvested from injured spine cord of rats were stained by double immunofluorescence labeling technique to observe the expression of NgR at histological and cellular levels. Macrophages which expressed NgR were constructed in vitro, and then the effects of NgR on macrophage phagocytosis and neuraxon regeneration in three groups (NgR-macrophages group, mock group and normal macrophages group) were studied using Western blot, micro-MTT colorimetry, and LDH assay separately. RESULTS: The results showed that CD68-positive macrophages in injured tissue of spine cord expressed NgR after double immunofluorescence staining on day 7 after SCI, and so did macrophages isolated and cultured from the injured spine cord. The results of Western blot showed that phagocytosis of macrophages in NgR-macrophages group was much better than that in mock group and normal macrophage group (p < 0.05). And the results of Micro-MTT colorimetry and LDH assay indicated that the capacity of neuraxon regeneration in NgR-macrophages group was significantly higher than that in the other two groups (p < 0.05). CONCLUSIONS: The results suggested that there was NgR expressing in the infiltrated macrophages following SCI, which increased phagocytosis of the macrophages, and promoted post-SCI CNS regeneration in vitro.

C2 translaminar screw as the optimal choice for atlantoaxial dislocation with C2-C3 congenital fusion.

OBJECTIVE AND IMPORTANCE: The entry point and trajectory are very important for transarticular screw (TAS) and C2 pedicle screw (PDS) plantation. When the physical size is not large enough for the screw passing through, an accurate entry point is the most important point for successful screw insertion without vertebral artery (VA) injury and spinal cord injury. Once the laminas of C2 and C3 are fused, the normal anatomic mark might disappear and the insertion point would be hard to find. As a result, the complication of TAS or PDS implantation increases rapidly. We used C2 translaminar screws (TLSs) with C1 lateral mass screws as the optimal fixation for atlantoaxial dislocation in order to reduce the risk of VA injury and spinal cord injury. CLINICAL PRESENTATION: A 37-year-old woman with atlantoaxial dislocation due to obsolete odontoid fracture complained of neck pain and myelopathy. Preoperative CT reconstruction showed C2-C3 fusion and small size of C2 isthmus. TECHNIQUE: The patient underwent posterior atlantoaxial fusion using C1 lateral mass screws and C2 TLSs. The posterior arch of atlas was removed for decompression and fusion was done at C1-C2 joints by grafting bone fragments from the posterior iliac crest. CONCLUSION: TLSs combined with C1 lateral mass screws might be a useful technique for patients with atlantoaxial dislocation and C2-C3 fusion, especially with small size of C2 isthmus. Also, the fusion of posterior elements between C2 and C3 might be a relative contraindication for TAS fixation.

Bioinformatics analyses of pathways and gene predictions in IL-1α and IL-1ß knockout mice with spinal cord injury.

PURPOSE: This study aimed to explore the potential genes and pathways regulated in spinal cord injury (SCI) model mice with IL-1α and IL-1ß knockout (KO). METHODS: Gene expression profile GSE70302, which includes data from injured spinal cord of 4 IL-1α-KO mice, 4 IL-1ß-KO mice and 4 C57BL with 6 mice as controls was downloaded from the Gene Expression Omnibus database. The differentially expressed genes (DEGs) of the IL-1α-KO or IL-1ß-KO vs. control, and IL-1α-KO vs. IL-1ß-KO groups were screened, followed by function enrichment and protein-protein interaction (PPI) analyses. Finally, miRNAs associated with SCI that may target the DEGs were predicted. RESULTS: A total of 579 and 992 DEGs were selected from the IL-1α-KO vs. control group and the IL-1ß-KO vs. control group, respectively, and 208 genes common between the 2 comparison groups were identified. Additionally, 526 DEGs were identified from the IL-1α-KO vs. IL-1ß-KO groups. These DEGs were significantly enriched in functions and pathways associated with ion transport, neuron apoptotic processes and inflammatory responses. The common genes were enriched in the pathways for cytokine-cytokine receptor interaction. DEGs of IL-1α-KO vs. IL-1ß-KO were significantly enriched in the immune system, hematopoietic cell lineage and PI3K-Akt signalling pathway-associated biological processes and pathways. The PPI network consisted of 76 nodes, such as Saa2, Kcna1, Scn8a, Ccl5, Ccl28 and Pink1. A total of 94 miRNAs, including mir-17-5P and mir-30a-5p were predicted that could target the DEGs. CONCLUSION: IL-1α and IL-1ß may play important roles in SCI by regulating ion transport, inflammation and neuron apoptotic processes and their associated genes or miRNAs. Compared with IL-1ß-KO, IL-1α-KO may improve the outcome of SCI via the alteration of hematopoietic cell lineage and PI3K-Akt signalling pathways.

Identification of critical genes in nucleus pulposus cells isolated from degenerated intervertebral discs using bioinformatics analysis.

Intervertebral disc (IVD) degeneration is a pathological process, which may lead to lower back pain. The present study aimed to investigate the pathogenesis of IVD degeneration. GSE42611 was downloaded from Gene Expression Omnibus, including 4 nucleus pulposus samples isolated from degenerated IVDs and 4 nucleus pulposus samples separated from normal IVDs. The differentially expressed genes (DEGs) between the degenerated and normal samples were screened using the limma package in R. Functional and pathway enrichment analyses were conducted separately for the upregulated and downregulated genes, using Database for Annotation, Visualization and Integrated Discovery software. In addition, protein­protein interaction (PPI) networks were constructed using the Search Tool for the Retrieval of Interacting Genes database and Cytoscape software. Finally, module analyses were conducted for the PPI networks using the MCODE plug­in in Cytoscape. A total of 558 DEGs were identified in the degenerated nucleus pulposus cells: 253 upregulated and 305 downregulated. Pathway enrichment analysis revealed that downregulated thrombospondin 1 (THBS1) was enriched in extracellular matrix­receptor interaction. Interleukin (IL)­6 in the PPI network for the upregulated genes and vascular endothelial growth factor A (VEGFA) in the PPI network for the downregulated genes had higher degrees. Additionally, four modules (µM1, µM2, µM3 and µM4) were identified from the PPI network for the upregulated genes. Four modules (dM1, dM2, dM3 and dM4) were identified from the PPI network for the downregulated genes. In the dM2 module, collagen genes and integrin subunit α4 (ITGA4) may interact with each other. Additionally, functional enrichment indicated that collagen genes were enriched in extracellular matrix organization. In conclusion, IL­6, VEGFA, THBS1, ITGA4 and collagen genes may contribute to the progression of IVD degeneration. These results suggested that the manipulation of these genes and their products may have potential as a novel therapeutic strategy for the treatment of patients with IVD.

[Pseudomonas aeruginosa elastase inhibits immune phagocytosis mediated by pulmonary surfactant protein A].

OBJECTIVE: To investigate the relationship between Pseudomonas aeruginosa (P.aeruginosa) elastase and pulmonary surfactant protein A (SP-A) in host infected by P.aeruginosa. METHODS: C3H mice were intranasally infected with P.aeruginosa wild-type PAO1, ΔlasB mutant and genetic complement strain PDO240LasB to determine the difference of virulence between wide type and mutant. The ability to degrade SP-A in vitro by PAO1, ΔlasB and PDO240LasB was observed through co-incubation of equal bacteria and SP-A and detected by Western blotting. The susceptibility of bacteria to phagocytosis was assayed by in vitro experiment that bacteria treated with SP-A was incubated with mouse Raw264.7 macrophages. RESULTS: Compared with wide-type PAO1, the virulence of ΔlasB mutant was attenuated in the mouse model of P.aeruginosa infection because of the knock down of elastase expression. The ΔlasB mutant lost the ability to degrade SP-A when incubated with SP-A in vitro. The in vitro phagocytosis experiments showed that SP-A augmented the phagocytosis of ΔlasB mutant bacteria more efficiently than the wild-type PAO1. CONCLUSION: P.aeruginosa elastase provides a protection from phagocytic cells by degrading SP-A.

Transarticular screw and C1 hook fixation for os odontoideum with atlantoaxial dislocation.

OBJECTIVE: To introduce a fixation device composed of C1-2 transarticular (TA) screws and C1 hooks and explore its indication and clinical outcome for os odontoideum with atlantoaxial dislocation (AAD). METHODS: From January 2004 to December 2008, 12 patients with os odontoideum (5 men and 7 women, average age 37.7 years [range 14-62 years]) were treated in the authors' hospital. All patients had AAD with local symptoms, and 10 had myelopathy. All patients underwent a posterior atlantoaxial fixation with C1-2 TA screws and C1 hooks. Clinical and radiographic analyses were performed at 3, 6, and 12 months postoperatively and annually thereafter. RESULTS: The follow-up period was 12-66 months (average follow-up period 35.5 months). No neurologic or vascular complications occurred in these cases, and the device was placed well with no loosening or breakage. Plain radiographs and three-dimensional reconstruction of computed tomography (CT) images revealed solid bony fusion with a good alignment of C1 and C2 at 3 months postoperatively. No hardware failure, pseudarthrosis, or instability was noted during the follow-up period. All patients had relief of pain within 3 months, and neurologic symptoms were substantially improved. CONCLUSIONS: When appropriate patients are selected, C1-2 TA screws combined with C1 hooks can be used to treat os odontoideum with AAD effectively with a relatively simple procedure resulting in excellent biomechanical strength and high bone fusion rate. Preoperative planning is crucial for the management of os odontoideum with AAD.

Bilateral atlantoaxial transarticular screws and atlas laminar hooks fixation for pediatric atlantoaxial instability.

STUDY DESIGN: An atlantoaxial fixation using bilateral C1-C2 transarticular screws and C1 laminar hooks was used in 5 pediatric patients, who were then followed up for 12 to 17 months to evaluate the technique. OBJECTIVE: To describe a modified posterior C1-C2 fixation technique and preliminary clinical and radiographic results in 5 pediatric patients. SUMMARY OF BACKGROUND DATA: Conventional posterior atlantoaxial fixations, such as Gallie and Brooks techniques, are frequently associated with high rates of pseudarthrosis and implant failure. The C1-C2 transarticular screw fixation has been shown to be effective in treatment of pediatric atlantoaxial instability, as well as adult atlantoaxial instability; however, this 2-point fixation merely stabilizes the atlantoaxial motion segment laterally. A 3-point fixation, composed with bilateral C1-C2 transarticular screws and C1 laminar hooks, has been developed. METHODS: Five patients with atlantoaxial instability, including 4 males and 1 female, aged 6 to 17 (average 10) years, underwent atlantoaxial fixation using bilateral C1-C2 transarticular screws and C1 laminar hooks during a 2-year period. The surgical technique and treatment procedures were intensively reviewed, and clinical symptoms and imaging appearance were retrospectively evaluated. RESULTS: Clinical follow-ups were obtained for an average of 14.4 (range: 12-17) months. The clinical and radiologic follow-up indicated a stable arthrodesis and offered clinical relief from symptoms for all patients. No neural or vascular impairment related to this technique was observed. CONCLUSION: Fixation of the atlantoaxial articulation using bilateral C1-C2 transarticular screws and C1 laminar hooks appears to be a reliable technique for treatment of pediatric atlantoaxial instability.