SSR1 and CKAP4 as potential biomarkers for intervertebral disc degeneration based on integrated bioinformatics analysis.

Background: Intervertebral disc degeneration (IDD) is a significant cause of low back pain and poses a significant public health concern. Genetic factors play a crucial role in IDD, highlighting the need for a better understanding of the underlying mechanisms. Aim: The aim of this study was to identify potential IDD-related biomarkers using a comprehensive bioinformatics approach and validate them in vitro. Materials and Methods: In this study, we employed several analytical approaches to identify the key genes involved in IDD. We utilized weighted gene coexpression network analysis (WGCNA), MCODE, LASSO algorithms, and ROC curves to identify the key genes. Additionally, immune infiltrating analysis and a single-cell sequencing dataset were utilized to further explore the characteristics of the key genes. Finally, we conducted in vitro experiments on human disc tissues to validate the significance of these key genes in IDD. Results: we obtained gene expression profiles from the GEO database (GSE23130 and GSE15227) and identified 1015 DEGs associated with IDD. Using WGCNA, we identified the blue module as significantly related to IDD. Among the DEGs, we identified 47 hub genes that overlapped with the genes in the blue module, based on criteria of |logFC| ≥ 2.0 and p.adj <0.05. Further analysis using both MCODE and LASSO algorithms enabled us to identify five key genes, of which CKAP4 and SSR1 were validated by GSE70362, demonstrating significant diagnostic value for IDD. Additionally, immune infiltrating analysis revealed that monocytes were significantly correlated with the two key genes. We also analyzed a single-cell sequencing dataset, GSE199866, which showed that both CKAP4 and SSR1 were highly expressed in fibrocartilage chondrocytes. Finally, we validated our findings in vitro by performing real time polymerase chain reaction (RT-PCR) and immunohistochemistry (IHC) on 30 human disc samples. Our results showed that CKAP4 and SSR1 were upregulated in degenerated disc samples. Taken together, our findings suggest that CKAP4 and SSR1 have the potential to serve as disease biomarkers for IDD.

LncRNA LINC00662 Exerts an Oncogenic Effect on Osteosarcoma by the miR-16-5p/ITPR1 Axis.

BACKGROUND: Osteosarcoma (OS) is one of the most malignant bone tumors and has a high metastatic rate. Increasing research has demonstrated the vital roles of long noncoding RNAs (lncRNAs) in human cancers, including OS. LncRNA LINC00662 has been revealed to act as an oncogene involved in multiple tumor progression. This study aimed to investigate the expression pattern, function, and regulatory mechanism of LINC00662 in OS. METHODS: Patients who underwent OS surgery were involved in this study. Experiments including RT-qPCR, MTT, western blot, FISH, RNA pull-down, luciferase reporter, colony formation, transwell invasion and migration, and sphere formation assay were performed to investigate the regulatory role of LINC00662 in OS. RESULTS: In the present study, our findings demonstrated the upregulation of LINC00662 expression in OS tissues and cells, and high expression of LINC00662 predicted a poor clinical prognosis of patients' iNOS. Through a series of in vivo assays, LINC00662 knockdown suppressed OS cell proliferation, invasion, migration, and stemness property maintenance. Further mechanistical investigations indicated that LINC00662 functioned as a competing endogenous RNA (ceRNA) for sponging microRNA-16-5p (miR-16-5p) to upregulate the expression of IP receptor type 1 (ITPR1) in OS cells. Restoration assays validated the involvement of ITPR1 in LINC00662-mediated regulation of cell functions in OS. CONCLUSION: LINC00662 exerts oncogenic functions in OS by targeting the miR-16-5p/ITPR1 axis.

Gene co-expression network analysis identifies BRCC3 as a key regulator in osteogenic differentiation of osteoblasts through a ß-catenin signaling dependent pathway.

OBJECTIVES: The prognosis of osteoporosis is very poor, and it is very important to identify a biomarker for prevention of osteoporosis. In this study, we aimed to identify candidate markers in osteoporosis and to investigate the role of candidate markers in osteogenic differentiation. MATERIALS AND METHODS: Using Weighted Gene Co-Expression Network analysis, we identified three hub genes might associate with osteoporosis. The mRNA expression of hub genes in osteoblasts from osteoporosis patients or healthy donor was detected by qRT-PCR. Using siRNA and overexpression, we investigated the role of hub gene BRCC3 in osteogenic differentiation by alkaline phosphatase staining and Alizarin red staining. Moreover, the role of ß-catenin signaling in the osteogenic differentiation was detected by using ß-catenin signaling inhibitor XAV939. RESULTS: We identified three hub genes that might associate with osteoporosis including BRCC3, UBE2N, and UBE2K. UBE2N mRNA and UBE2K mRNA were not changed in osteoblasts isolated from osteoporosis patients, compared with healthy donors, whereas BRCC3 mRNA was significantly increased. Depletion of BRCC3 promoted the activation of alkaline phosphatase and formation of calcified nodules in osteoblasts isolated from osteoporosis patients and up-regulated ß-catenin expression. XAV939 reversed the BRCC3 siRNA-induced osteogenic differentiation. Additionally, inhibited osteogenic differentiation was also observed after BACC3 overexpression, and this was accompanied by decreased ß-catenin expression. CONCLUSION: BRCC3 is an important regulator for osteogenic differentiation of osteoblasts through ß-catenin signaling, and it might be a promising target for osteoporosis treatment.

Inhibitory Effect of MicroRNA-107 on Osteosarcoma Malignancy Through Regulation of Wnt/ß-catenin Signaling in Vitro.

We showed that miR-107 expression was decreased in osteosarcoma (OS) tissues and cell lines. miR-107 mimic significantly decreased OS cell proliferation and inhibited invasion and migration of OS cells. Inhibition of miR-107 expression notably promoted proliferation, invasion and migration of OS cells. In addition, miR-107 mimic inhibited EMT biomarkers and significantly increased apoptosis. miR-107 mimic significantly decreased the protein expression of ß-catenin, Cyclin D1, and c-Myc, whereas increased GSK-3ß protein expression. miR-107 mimic markedly reduced the luciferase activity of 3'UTR of ß-catenin. Overexpression of ß-catenin inhibited miR-107 mimic-induced decrease of cell proliferation, invasion and migration ability, and increase of apoptosis.

Outcomes of autograft alone versus PEEK+ autograft interbody fusion in the treatment of adult lumbar isthmic spondylolisthesis.

OBJECTIVES: Bone resulting from a complete resection of the posterior arch can be cut into an autograft bone that contains the facet joint structure and morselised bone for interbody fusion. However, whether a strut autograft that contains this trimmed facet joint can produce the same clinical and radiographic outcomes as a cage for interbody fusion remains unclear. The aim of this study was to compare the outcomes of a local facet joint autograft alone to those of polyetheretherketone (PEEK)+autograft for posterior lumbar interbody fusion (PLIF) in the treatment of adult isthmic spondylolisthesis. PATIENTS AND METHODS: A retrospective analysis was performed on 84 patients with single lumbar isthmic spondylolisthesis who were treated with a local facet joint autograft alone (group A; n=44) or PEEK+autograft (group B; n=40) in PLIF with a minimum follow-up period of 24 months. Pain and disability were assessed using the visual analogue scale, Oswestry disability index and Kirkaldy-Willis criteria. In the radiological evaluation, disc height, slippage reduction, and fusion status were examined. Postoperative complications were also monitored. RESULTS: At the last follow-up examination, 84.1% (37/44) of the patients in group A and 82.5% (33/40) of the patients in group B had a good outcome, and there were no significant differences between the two groups. Boh Methods led to significant improvements in disc height, and while PEEK+autograft produced a smaller loss in disc height, the difference was insignificant. The improvements in slippage and the fusion and complication rates between the two groups were similar. CONCLUSION: There were no significant differences in the clinical outcomes or radiographic improvements of both fusion methods in the treatment of adult isthmic spondylolisthesis. An autograft excised from a complete posterior arch containing a facet joint for interbody fusion is effective and affordable for treating isthmic spondylolisthesis.

Easy Preparation and Photoelectrochemical Properties of CdS Nanoparticle/Graphene Nanosheet Nanocomposites Using Supercritical Carbon Dioxide.

Graphene nanosheets (GNSs) were modified with CdS nanoparticles (NPs) using supercritical CO2 (SC CO2), which has gas-like diffusivity, low viscosity, and near-zero surface tension. The resulting CdS NP/GNS nanocomposites were characterized by field-emission scanning electron microscopy, transmission electron microscopy, X-ray diffraction, X-ray photoelectron spectroscopy, Raman spectroscopy, and photoluminescence spectroscopy. Distinct morphologies of CdS NP/GNS nanocomposites decorated on the GNS surface were obtained at different SC CO2 pressures, temperatures, and durations and in different sources. Results showed that the sources and SC CO2 significantly influenced the aggregation or assembly behavior of the CdS NP/GNS nanocomposites on the GNSs. The formation mechanism of the distinct nanohybrid structures was studied by Raman mapping. A difference was noted between the Raman spectra of pristine graphene nanosheets and CdS NP/GNS nanocomposites. This result can be ascribed to the CdS NPs anchored onto the GNS defects and to the improved quality of the GNSs under SC CO2. The photo-current densities of CdS NP/GNS nanocomposites were at least three times higher than that of the pristine CdS NPs at the same applied voltage for photoelectrochemical water splitting. The findings suggested that highly efficient graphene-supported NP photoelectrocatalysts can be fabricated by the supercritical fluid method and that graphene can serve as a favorable photoelectrocatalytic carrier, with promising potential applications in environmental and energy fields. Keywords: Graphene Nanosheets, Cadmium Sulfide, Raman Spectroscopy, Photoelectrochemical.

[Comparison of inter- and intra-observer reliability between GATA and SMU classification systems for spinal tuberculosis].

OBJECTIVE: To compare the inter- and intra-observer reliability of the GATA and SMU classification systems for spinal tuberculosis and assess the clinical value of SMU classification. METHODS: One hundred patients with spinal tuberculosis treated in our hospital from January 2004 to December 2011 were randomly selected for analysis, including 54 males and 46 females with a mean age of 45 years (range, 16-68 years). All the patients had X-ray, CT and MRI examinations. Five observers experienced in spinal tuberculosis independently assigned the classification using the GATA and SMU classification systems, and the assignment was repeated 3 months later to test its reproducibility. Kappa value was used to determine the intra- and inter-observer reliability. RESULTS: For GATA and SMU classification systems, the inter-observer percentage of agreement averaged (59.9∓4.84)% (κ=0.412∓0.058) and (81.6∓6.06)% (κ=0.753∓0.068), and the intra-observer percentage of agreement was (75.6∓5.27)% (κ=0.624∓0.078) and (89.8∓2.28)% (κ=0.862∓0.037), respectively. CONCLUSION: The SMU classification system of spinal tuberculosis has a higher inter-observer and intra-observer reliability than the GATA classification system, but its clinical value needs to be further tested in future clinical trials.

[Jiawei shentong zhuyu decoction prevented the occurrence of failed back surgery syndrome and its effect on serum TNF-alpha a clinical study].

OBJECTIVE: To explore the clinical roles of Jiawei Shentong Zhuyu Decoction (JSZD) in preventing the occurrence of failed back surgery syndrome (FBSS), and to observe its effect on serum tumor necrosis factor-alpha (TNF-alpha). METHODS: Totally 100 patients prepared for surgical operation due to lumbar intervertebral disc herniation were randomly assigned to the treatment group and the control group according to random number table, 50 cases in each group. Patients in the treatment group additionally took JSZD, one dose per day, taken in two portions, once in the morning and once in the evening. Those in the control group took Celecoxib Capsule (200 mg each time, once per day) and Mecobalamin Tablet (0.5 mg each time, 3 times per day). They only took Mecobalamin Tablet from the 11th day. All patients were treated for 30 days. Japanese Orthopaedic Association (JOA) score was performed before treatment, at week 1, after treatment, at 6 months of followed-ups, and at 12 months of followed-ups. And the levels of TNF-alpha in the peripheral blood were observed before treatment and at one month after treatment. RESULTS: Totally 93 patients completed the followed-up study. The JOA scores were improved after treatment, at 6 and 12 months of followed-ups (P < 0.05, P < 0.01). The JOA score at 6 months of followed-ups was superior in the treatment group to that of the control group (P < 0.05). Five patients (accounting for 10.6%) suffered from FBSS in the treatment group, while 9 (accounting for 19.6%) suffered from FBSS in the control group. The treatment group was superior to the control group (P < 0.05). The TNFalpha level was improved after treatment in the two groups. Of them, the improvement of TNF-alpha in the treatment group was better than that of the control group (P < 0.05). CONCLUSION: The application of JSZD was effective for preventing the occurrence of FBSS, and improved the serum TNF-alpha level.

[Clinical study of transforaminal instrumented lumbar interbody fusion].

OBJECTIVE: To study the effect and complications of transforaminal lumbar interbody fusion technique. METHODS: The medical records and radiographs of 40 patients undergone transforaminal lumbar interbody fusion between 2005 and 2007 were retrospectively reviewed. There were 49 segments with fusion. Preoperative and 1 year postoperative functional evaluation were graded with ODI and VAS scoring system. The height and angle of the intervertebral space and the fusion status were measured as well. RESULTS: All patients were followed-up for 12 to 24 months with the average of 18 mouths. There were no severity postoperative complications. The operation time averaged 160 min and average blood loss 510 ml. The effect results were excellent in 28 cases, good in 7 and fair in 5. The fusion rate was 100%. One year after operation, the pain relief in the VAS and the reduction of the ODI were significant (P < 0.01), the height and angle of the intervertebral space increased obviously (P < 0.05). Fifteen patients complained low back pain to some extent untill the last follow-up. CONCLUSION: Transforaminal lumbar interbody fusion can achieve satisfactory clinical and radiographic results especially for the failed back surgery syndrome.

[The synthetic typing and its clinical application in atlantoaxial dislocation].

OBJECTIVE: To evaluate the synthetic typing and the treatment strategy for atlantoaxial dislocation. METHODS: The synthetic typing of atlantoaxial dislocation was worked out on the base of pathogenesis typing, Fielding imaging typing, and clinical typing, named PIR typing system (Pathogenesis, Imaging, and Reduction). Ninety-three patients with atlantoaxial dislocation were treated according to this typing system. RESULTS: Nine cases of type-II dens fracture were treated with hollow screw fixation. Bone union was accomplished at the follow-up of three months in all the patients, only with slight limitation of cervical motion. Un-retrieved Fielding I -degree dislocation was found in one case. Among the thirty-four patients treated with trans-oropharyngeal atlantoaxial reduction plate system (TARP), 32 obtained complete atlantoaxial reduction and fusion three months after operation. Atlantoaxial dislocation recurred in the other two cases because of screw loosening and the problem was solved through revision operations. Four patients in non-reducible type underwent anterior and/or posterior decompression. T heir neurological improved after operation but their atlantoaxial joints remained dislocated, and one case complicated with intracranial infection. CONCLUSIONS: Via the synthetic PIR typing system, atlantoaxial dislocation can be better classified according to its pathogenesis, imaging manifestation and mechanic stability. This system can also be served as a guide for clinical treatment. Anterior TARP operation and posterior atlantoaxial trans-pedicle screw-rod fixation are the main methods for the treatment of atlantoaxial dislocation.

[The reason and prevention of upper cervical reoperations].

OBJECTIVE: To discuss the reasons for the operation performed on 13 patients with upper cervical disease and to explore the management and prevention of upper cervical disease. METHODS: Thirteen patients with upper cervical disease were retrospectively reviewed. The reason for of reoperations on these patients were analyzed. The measures to reduce upper cervical operational complication and bad prognosis were discussed to avoid reoperations. RESULTS: The reasons for reoperations included 9 cases with unstable or re-dislocated atlantoaxial joint, 10 cases with residual spinal cord compression, 1 case with malposition of odontoid screw, 1 case with adjacent cervical spine regression, 1 case with occipital-cervical fusion failure, 1 case with spinal cord injury during operation, 1 case with bone-plant slipped into canales spinalis, and 1 case with demand to take out internal fixation for aggravated symptom. CONCLUSIONS: The common reasons for upper cervical reoperations were due to instability or redislocation of atlantoaxial joint and residual of spinal cord compression. Some measures such as reducing operate miss, using firm internal fixation and decompressing were advisable to decrease the incidence of reoperations.

[Mosaicplasty osteochondral grafting to repair cartilaginous defects under arthroscopy].

OBJECTIVE: To study mosaicplasty a as method of autogenous osteochondral transplantation in the treatment of cartilaginous defects. METHODS: The technique involves obtaining small cylindrical grafts from the non-weight bearing periphery of the femur at the patellar femoral joint, and transporting them to the prepared recipient site by arthroscopy. RESULTS: Fifteen patients with defects cartilaginous received mosaicptasty osteochondral grafting. Follow up for 12 to 21 months (mean 15 months) showed good results. CONCLUSION: The treatment is indicated for patients with focal cartilaginous defects under the age of 45.