Forsythoside B attenuates neuro-inflammation and neuronal apoptosis by inhibition of NF-κB and p38-MAPK signaling pathways through activating Nrf2 post spinal cord injury.

BACKGROUND: Spinal cord injury (SCI) is a ruinous neurological pathology that results in locomotor and sensory impairment. Neuro-inflammation and secondary neuronal apoptosis contribute to SCI, with anti-inflammatory therapies the focus of many SCI studies. Forsythoside B (FTS•B), a phenylethanoid glycoside extracted from the leaves of Lamiophlomis rotata Kudo, has been shown previously to have anti-inflammatory properties. Nevertheless, the therapeutic effect of FTS•B on neuro-inflammation after SCI is unknown. METHODS: Neuro-inflammation was assessed by western blotting (WB), immunofluorescence (IF) staining, and enzyme-linked immunosorbent assay (ELISA) both in vitro and in vivo. Secondary neuronal apoptosis was simulated in a microglia-neuron co-culture model with the degree of apoptosis measured by WB, IF, and TUNEL staining. In vivo, FTS•B (10 mg/kg, 40 mg/kg) were intraperitoneally injected into SCI mice. Morphological changes following SCI were evaluated by Nissl, Hematoxylin-eosin, and Luxol Fast Blue staining. Basso Mouse Scale scores were used to evaluate locomotor function recovery. RESULTS: FTS•B markedly decreased the levels of iNOS, COX-2 and signature mediators of inflammation. Phosphorylated p38 and nuclear factor-kappa B (NF-κB) were markedly decreased by FTS•B. Additionally, FTS•B-induced inhibition of NF-κB and p38-MAPK signaling pathways was reversed by Nrf2 downregulation. Administration of FTS•B also significantly reduced apoptosis-related protein levels indicating that FTS•B ameliorated secondary neuronal apoptosis. FTS•B administration inhibited glial scar formation, decreased neuronal death, tissue deficiency, alleviated demyelination, and promoted locomotor recovery. CONCLUSION: FTS•B effectively attenuates neuro-inflammation and secondary neuronal apoptosis by inhibition of NF-κB and p38-MAPK signaling pathways through activating Nrf2 after SCI. This study demonstrates FTS•B to be a potential therapeutic for SCI.

Growth Differentiation Factor 15 Regulates Oxidative Stress-Dependent Ferroptosis Post Spinal Cord Injury by Stabilizing the p62-Keap1-Nrf2 Signaling Pathway.

Background: Spinal cord injury (SCI) is a severe traumatic disorder of the central nervous system (CNS) that causes irreversible damage to the nervous tissue. The consequent hemorrhage contributed by trauma induces neuronal ferroptosis post SCI, which is an important death mode to mediate neuronal loss. Growth differentiation factor 15 (GDF15) is a cytokine that regulates cell proliferation, differentiation, and death. However, the specific role of GDF15 in neuronal ferroptosis post SCI remains unknown. Materials and Methods: Neuronal ferroptosis in vitro was measured by detection of lipid peroxidation, glutathione, iron content, and reactive oxidative stress. In vivo, western blotting and immunofluorescence (IF) staining was utilized to measure ferroptosis post SCI. IF staining, TUNEL staining, hematoxylin-eosin staining, and Nissl staining were used to measure neurological damage. Finally, locomotor function recovery was analyzed using the Basso Mouse Scale and Louisville Swim Scale. Results: GDF15 was significantly increased in neuronal ferroptosis and silencing GDF15 aggravated ferroptosis both in vitro and in vivo. Besides, GDF15-mediated inhibition of neuronal ferroptosis is through p62-dependent Keap1-Nrf2 pathway. In SCI mice, knockdown of GDF15 significantly exacerbated neuronal death, interfered with axon regeneration and remyelination, aggravated ferroptosis-mediated neuroinflammation, and restrained locomotor recovery. Conclusion: GDF15 effectively alleviated neuronal ferroptosis post SCI via the p62-Keap1-Nrf2 signaling pathway and promoted locomotor recovery of SCI mice, which is suggested as a potential target on SCI pathogenesis and treatment.

FANCC deficiency mediates microglial pyroptosis and secondary neuronal apoptosis in spinal cord contusion.

BACKGROUND: Traumatic spinal cord injury (SCI)-induced neuroinflammation results in secondary neurological destruction and functional disorder. Previous findings showed that microglial pyroptosis plays a crucial role in neuroinflammation. Thus, it is necessary to conduct a comprehensive investigation of the mechanisms associated with post-SCI microglial pyroptosis. The Fanconi Anemia Group C complementation group gene (FANCC) has been previously reported to have an anti-inflammation effect; however, whether it can regulate microglial pyroptosis remains unknown. Therefore, we probed the mechanism associated with FANCC-mediated microglial pyroptosis and neuroinflammation in vitro and in vivo in SCI mice. METHODS: Microglial pyroptosis was assessed by western blotting (WB) and immunofluorescence (IF), whereas microglial-induced neuroinflammation was evaluated by WB, Enzyme-linked immunosorbent assays and IF. Besides, flow cytometry, TdT-mediated dUTP Nick-End Labeling staining and WB were employed to examine the level of neuronal apoptosis. Morphological changes in neurons were assessed by hematoxylin-eosin and Luxol Fast Blue staining. Finally, locomotor function rehabilitation was analyzed using the Basso Mouse Scale and Louisville Swim Scale. RESULTS: Overexpression of FANCC suppressed microglial pyroptosis via inhibiting p38/NLRP3 expression, which in turn reduced neuronal apoptosis. By contrast, knockdown of FANCC increased the degree of neuronal apoptosis by aggravating microglial pyroptosis. Besides, increased glial scar formation, severe myelin sheath destruction and poor axon outgrowth were observed in the mice transfected with short hairpin RNA of FANCC post SCI, which caused reduced locomotor function recovery. CONCLUSIONS: Taken together, a previously unknown role of FANCC was identified in SCI, where its deficiency led to microglia pyroptosis, neuronal apoptosis and neurological damage. Mechanistically, FANCC mediated microglia pyroptosis and the inflammatory response via regulating the p38/NLRP3 pathway.

LncRNA-COX2 inhibits Fibroblast Activation and Epidural Fibrosis by Targeting EGR1.

Rationale: Epidural fibrosis is one of the contributors to failed back surgery syndrome (FBSS) with a high incidence of about 80,000 cases per year. The fibrosis spreads from the operative region to the dura mater or the nerve root and results in functional incapacity and pain after laminectomy. Our previous study showed that down-regulation of lncRNA-COX2 is involved in the epidural scar formation. However, it remains unknown whether lncRNA-COX2 participate in the fibroblast activation and epidural fibrogenesis. Methods: LncRNA-COX2 and EGR1 expression were assessed by qRT-PCR and western blotting. Fibroblasts differentiation, proliferation and migration was determined by Collagen I/ɑ-SMA, 5-ethynyl-2'-deoxyuridine (EdU) and Transwell Assay respectively. Luciferase reporter assay was performed for the verification of target of LncRNA-COX2. Laminectomy was performed to establish the model of epidural fibrosis in mice. Epidural scar was evaluated by hematoxylin and eosin (HE) staining and Masson Trichrome staining. Results: Based on the result of transcriptome profiling, we found LncRNA-COX2 was significantly decreased in epidural tissues after laminectomy and in activated fibrotic fibroblasts. In vitro, overexpression of LncRNA-COX2 suppressed epidural fibrogenesis by inhibiting fibroblasts differentiation, proliferation and migration. Mechanistically, LncRNA-COX2 functioned as competing endogenous RNA (ceRNA) of EGR1. Gain of LncRNA-COX2 significantly decreased the expression of EGR1 and showed anti-fibrotic effect while EGR1 was markedly increased after loss of LncRNA-COX2. In vivo, LncRNA-COX2 attenuated laminectomy-induced epidural fibrosis in mice. Conclusion: In summary, the results demonstrated that LncRNA-COX2 showed anti-fibrotic effect by targeting EGR1 and identified LncRNA-COX2 as therapeutic molecule for preventing aberrant epidural fibrosis.

Activation of glucagon-like peptide-1 receptor in microglia attenuates neuroinflammation-induced glial scarring via rescuing Arf and Rho GAP adapter protein 3 expressions after nerve injury.

Rationale: The neuroinflammation is necessary for glial group initiation and clearance of damaged cell debris after nerve injury. However, the proinflammatory polarization of excessive microglia amplifies secondary injury via enhancing cross-talk with astrocytes and exacerbating neurological destruction after spinal cord injury (SCI). The glucagon-like peptide-1 receptor (GLP-1R) agonist has been previously shown to have a neuroprotective effect in neurodegeneration, whereas its potency in microglial inflammation after SCI is still unknown. Methods: The effect and mechanism of GLP-1R activation by exendin-4 (Ex-4) were investigated in in vitro cultured glial groups and in vivo in SCI mice. Alterations in the gene expression after GLP-1R activation in inflammatory microglia were measured using mRNA sequencing. The microglial polarization, neuroinflammatory level, and astrocyte reaction were detected by using western blotting, flow cytometry, and immunofluorescence. The recoveries of neurological histology and function were also observed using imaging and ethological examinations. Results: GLP-1R activation attenuated microglia-induced neuroinflammation by reversing M1 subtypes to M2 subtypes in vitro and in vivo. In addition, activation of GLP-1R in microglia blocked production of reactive astrocytes. We also found less neuroinflammation, reactive astrocytes, corrected myelin integrity, ameliorated histology, and improved locomotor function in SCI mice treated with Ex-4. Mechanistically, we found that Ex-4 rescued the RNA expression of Arf and Rho GAP adapter protein 3 (ARAP3). Knockdown of ARAP3 in microglia reversed activation of RhoA and the pharmacological effect of Ex-4 on anti-inflammation in vitro. Conclusion: Ex-4 exhibited a previously unidentified role in reducing reactive astrocyte activation by mediation of the PI3K/ARAP3/RhoA signaling pathway, by neuroinflammation targeting microglia, and exerted a neuroprotective effect post-SCI, implying that activation of GLP-1R in microglia was a therapeutical option for treatment of neurological injury.

LINC00266-1/miR-548c-3p/SMAD2 feedback loop stimulates the development of osteosarcoma.

Osteosarcoma (OS) is one of the most common primary bone malignancies and accounts for 3.4% of pediatric tumors. Its 5-year survival is as low as about 20%. Differentially expressed lncRNAs in OS profiling were searched in the downloaded profile of GSE12865. As a result, LINC00266-1 was detected to be upregulated in both GSE12865 and OS tissues we collected. SMAD2 was the downstream target binding to promoter sites of LINC00266-1, displaying a positive regulatory interaction. Knockdown of LINC00266-1 suppressed the proliferative and metastatic abilities, and promoted the apoptosis in OS cells. Besides, knockdown of LINC00266-1 significantly alleviated the growth of OS in vivo. MiR-548c-3p was the sponge miRNA of LINC00266-1, which was able to reverse the regulatory effects of LINC00266-1 on OS cell phenotypes. Moreover, miR-548c-3p bound to the 3'-UTR of SMAD2 and thus downregulated SMAD2. Overexpression of SMAD2 partially reversed the regulatory effects of LINC00266-1 on OS cell phenotypes. Finally, we have identified that LINC00266-1/miR-548c-3p/SMAD2 feedback loop was responsible for stimulating the development of OS.

Comparison of the Effects of Proximal Humeral Internal Locking System (PHILOS) Alone and PHILOS Combined with Fibular Allograft in the Treatment of Neer Three- or Four-part Proximal Humerus Fractures in the Elderly.

OBJECTIVE: To compare and analyze the clinical outcomes of the proximal humeral internal locking system (PHILOS) alone and the PHILOS combined with fibular allograft in the treatment of Neer three- and four-part proximal humerus fractures (PHF) in the elderly. METHODS: From January 2014 to January 2018, a total of 42 elderly patients with Neer three- or four-part PHF admitted to our hospital were randomly divided into observation group and control group, with 21 patients in each group. The observation group was treated with the PHILOS combined with fibular allograft. The control group was treated with the PHILOS alone. Perioperative parameters and fracture classification were recorded in the two groups. Function results were assessed by Visual Analog Scale (VAS), Constant-Murley score (CMS), American Shoulder and Elbow Surgeons (ASES) score, and the Disability of Arm-Shoulder-Hand (DASH) score. Radiological results were evaluated using the neck-shaft angle (NSA) and humeral head height (HHH), and complications were also recorded in each group. RESULTS: There were no significant differences between the two groups in terms of preoperative status, age, gender, cause of trauma, fracture site, and fracture classification. The average follow-up time was 12 months. At the last follow-up, the VAS and DASH observation groups were lower than the control group, and there was significant difference between the two groups (P < 0.05). The CMS and ASES were higher in the observation group than the control group, and there was significant difference between the two groups (P < 0.05). The mean difference in the NSA and HHH were lower in the observation group than the control group, and there was a significant difference between the two groups (P < 0.05). There was one postoperative complication in the observation group, which was humeral head avascular necrosis (AVN). There were seven postoperative complications in the control group, including three cases of humeral head collapse and three cases of screw cutout and one case of humeral head AVN. The incidence of postoperative complications in the observation group was significantly lower than the control group (P < 0.05), there was a significant difference between the two groups. CONCLUSIONS: For Neer three- or four-part PHF in the elderly patients, PHILOS fixation with fibular allograft shows satisfactory short-term results with respect to humeral head support and maintenance of reduction, and may reduce the incidence of complications associated with fixation using a PHILOS alone.

CircRNA LRP6 promotes the development of osteosarcoma via negatively regulating KLF2 and APC levels.

We aimed to investigate the biological functions of circLRP6 in the progression of osteosarcoma. CircLRP6 level in OS was detected by quantitative real-time polymerase chain reaction. Correlation between circLRP6 level with survival of OS patients was evaluated. Cell counting kit-8 and Transwell assay were conducted to detect proliferative, migratory and invasive capacities of OS cells. Cell cycle and apoptosis in OS cells influenced by circLRP6 were evaluated by flow cytometry. RNA immunoprecipitation was conducted to verify the binding relationship between circLRP6 with LSD1 and EZH2. Finally, the interaction between LSD1, EZH2 and promoter regions of KLF2, APC was clarified by chromatin immunoprecipitation. CircLRP6 level markedly increased in OS tissues. Besides, OS patients with high expression of circLRP6 showed shorter disease-free survival and over-all survival than those with low expression. CircLRP6 knockdown suppressed proliferative, migratory and invasive rates of OS cells. Moreover, circLRP6 knockdown induced apoptosis and arrested cell cycle in G0/G1 phase. The interaction between circLRP6 with LSD1 and EZH2 mediates their binding to the promoter regions of KLF2 and APC. Knockdown of circLRP6 weakened the binding abilities of LSD1, EZH2 to KLF2, APC. APC overexpression inhibited proliferation, induced apoptosis and arrested cell cycle. Moreover, the tumor-suppressor effect of downregulated circLRP6 on OS could be reversed by APC knockdown. Collectively, circLRP6 was highly expressed in OS and served as an oncogene by binding to LSD1 and EZH2 to inhibit expressions of KLF2 and APC.

Hsa-miR-889-3p promotes the proliferation of osteosarcoma through inhibiting myeloid cell nuclear differentiation antigen expression.

Osteosarcoma accounts for about 0.2% in human malignant solid tumors. The mortality and metastatic rates of osteosarcoma remain relatively high. MicroRNA (miRNA) is a kind of non-coding small-molecular RNA discovered in recent years. Various studies have identified the involvement of miRNA in the occurrence and development of tumor as an oncogene or tumor-suppressor gene. This study aims to investigate the effect of hsa-miR-889-3p on the progression of osteosarcoma and its underlying mechanism. Through the bioinformatics methods, we first found that hsa-miR-889-3p was upregulated in osteosarcoma, and it was a prognostic risk factor for osteosarcoma. Additionally, the gene set enrichment analysis (GSEA) revealed that hsa-miR-889-3p mainly affected cell cycle progression and proliferation of osteosarcoma. To verify the bioinformatics results, regulatory effects of hsa-miR-889-3p on osteosarcoma both in vitro and in vivo experiments were investigated. It is found that hsa-miR-889-3p could promote the proliferation of osteosarcoma cells though regulating cell cycle progression. Tumor size and growth rate of osteosarcoma were influenced by hsa-miR-889-3p in xenograft models. To further explore its potential mechanism, the target gene of hsa-miR-889-3p was predicted. Furthermore, hsa-miR-889-3p was confirmed to inhibit the expression of myeloid cell nuclear differentiation antigen (MNDA) in a targeted manner. In conclusion, hsa-miR-889-3p could promote the proliferation of osteosarcoma through inhibiting MNDA expression, which provides a potential therapeutic strategy in treatment for osteosarcoma.

[Case control study on Zero-profile intervertebral fusion system and conventional cage-plate intervertebral fusion system for the treatment of multi-segment cervical spondylosis].

OBJECTIVE: To explore the clinical effect in the near future between Zero-profile intervertebral fusion system (Zero-P) and conventional cage-plate intervertebral fusion system (CCP) for the multi-segment(>=2 segments) cervical spondylosis. METHODS: Forty-two patients with cervical spondylosis who underwent multi-segment decompression of the cervical spine from October 2012 to October 2017 were selected as subjects. Zero-P was applied in 21 patients (Zero-P group) and CCP was applied in 21 patients(CCP group). The general condition and perioperative parameters of all the patients were recorded. VAS, JOA scores and incidence of dysphagia were observed before and after operation. The prevertebral soft tissue thickness was measured at 1 week, 1 month after operation and at the last follow-up. At the same time, the Cobb angle of the functional unit of the fusion segments was measured, and the overall curvature change of the cervical vertebra was observed. The clinical efficacy was reviewed at 1 week, 1, 3, 12 months after surgery, and the AP and lateral cervical X-rays were reviewed to evaluate the internal fixation effect. RESULTS: There were no significant differences in age, gender, duration of disease, surgical segment, follow-up time and hospitalization time between two groups(P>0.05). The length of the surgical incision, intraoperative blood loss, operation time, postoperative drainage volume in the Zero-P group were(4.37±0.72) cm, (50.9±7.98)ml, (84.4±8.18) min, (76.2±10.13) ml, respectively, and those in CCP group were (6.50±0.71) cm, (108.6±9.25) ml, (118.6±8.55) min, (130.1±9.42) ml, respectively. There were signigicant differences in above items between two groups(P<0.05). There were no significant difference in the VAS and JOA improvement rate between two groups at the last follow-up (P>0.05). There was no significant difference in the overall physiological curvature of the cervical vertebra between two groups (P>0.05). The prevertebral soft tissue thickness at 1 week, 1 month after operation, final follow-up respectively was(11.6±1.9), (9.8±1.4), (9.5±1.6) mm in Zero-P group, and in CCP group those were(12.5±2.6), (11.1±2.4), (11.0±1.9) mm, respectively. There were significant differences in each time point between two groups(P<0.05). At the last follow-up, no complication of dysphagia was found in Zero-P group, and three patients had dysphagia in CCP group, with a statistically significant difference between two groups (P<0.05). CONCLUSIONS: Multiple segmental decompression fusion to treat cervical spondylosis, regardless of the traditional CCP system or new Zero-P system are effective methods, but the Zero-P system has advantages of small surgical incision, short operation time, less intraoperative bleeding, convenient operation, better recovery of prevertebral soft tissue swelling, and lower possibility of postoperative dysphagia.

LncRNA SNHG3/miRNA-151a-3p/RAB22A axis regulates invasion and migration of osteosarcoma.

To elucidate the potential function of lncRNA SNHG3 in the development of osteosarcoma. Quantitative real-time polymerase chain reaction was conducted for detection of SNHG3, miRNA-151a-3p and RAB22 A in osteosarcoma tissues and cells. Receiver operating characteristic curve was introduced to analyze the diagnostic potential of SNHG3 in osteosarcoma. Correlation between SNHG3 expression and the overall survival of osteosarcoma patients was evaluated using Kaplan-Meier method. Invasive and migratory potentials of osteosarcoma cells were examined by Transwell assay. Furthermore, dual-luciferase reporter gene assay, RNA-pull down and RIP assay were used to verify the binding of SNHG3/RAB22 A to miRNA-151a-3p. The function of SNHG3/miRNA-151a-3p/RAB22 A axis in osteosarcoma was finally confirmed by rescue experiments. SNHG3 and RAB22 A were highly expressed in osteosarcoma patients, while miRNA-151a-3p was lowly expressed. The overall survival of osteosarcoma patients with high expression of SNHG3 was shorter than those with low expression. SNHG3 overexpression markedly promoted invasive and migratory potentials of osteosarcoma cells. Through dual-luciferase reporter gene assay, both SNHG3 and RAB22 A could bind to miRNA-151a-3p. RAB22 A expression was positively regulated by SNHG3, but negatively regulated by miRNA-151a-3p. Finally, rescue experiments confirmed that RAB22 A overexpression could reverse the promotive effects of miRNA-151a-3p knockdown on invasive and migratory potentials of osteosarcoma cells. SNHG3 is highly expressed in osteosarcoma, and promotes the invasive and migratory potentials of osteosarcoma cells by absorbing miRNA-151a-3p to upregulate RAB22 A expression.

Long non-coding RNA XIST promotes osteoporosis through inhibiting bone marrow mesenchymal stem cell differentiation.

The purpose of the present study was to identify the key long non-coding (lnc)RNAs in the occurrence and development of osteoporosis (OP) and to explore the associated molecular mechanism. First, the Gene Expression Omnibus (GEO) datasets, with key words 'osteoporosis' and 'HG-133A', were screened. RankProd R package was used to calculate the dysregulated lncRNAs in OP. Following this, bone marrow mesenchymal stem cells (BM-MSCs) harvested from 3-week-old Sprague Dawley rats were employed for detection of osteoblast differentiation. Following overexpression or interference with X-inactive specific transcript (XIST), osteogenesis-associated genes and proteins in BM-MSCs were detected using reverse transcription-quantitative polymerase chain reaction and western blot analysis. Alkaline phosphatase (ALP) and Alizarin Red S staining were also performed to measure the osteogenic ability of BM-MSCs. Results from the two datasets indicated that 6 lncRNAs were dysregulated in OP. Notably, XIST is key lncRNA in diverse diseases, and was subsequently selected for analysis. It was revealed that XIST was significantly upregulated in plasma and monocytes from patients with OP compared with the normal controls. Furthermore, results indicated that overexpression of XIST significantly inhibited osteoblast differentiation in BM-MSCs, as evidenced by the decreased expression of ALP, bone γ-carboxyglutamic acid-containing protein and runt related transcription factor 2, reduced ALP activity and a decreased number of calcium deposits. However, interference of XIST exhibited the opposite biological effects in BM-MSCs. Taken together, XIST was highly expressed in the serum and monocytes of patients with OP. In addition, the findings suggested that XIST could inhibit osteogenic differentiation of BM-MSCs.

miRNA-544a Regulates the Inflammation of Spinal Cord Injury by Inhibiting the Expression of NEUROD4.

BACKGROUND/AIMS: To explore the potential role of miR-544a in spinal cord injury and the possible mechanism involved. METHODS: We established a mouse model with spinal cord injury to examine the changes in grip force recovery of the forelimb or the posterior limb of the mouse. Microarray was performed to achieve differentiated miRNAs in the mice. The expressions of miR-544a, MCP-1, IL36B and IL17B after spinal cord injury were detected by qRT-PCR. Subsequently, miR-544a was overexpressed to observe changes in inflammation and grip strength after spinal cord injury. Target gene of miR-544a was then predicted using bioinformatics technology. Finally, dual luciferase reporter gene assay was used to verify the binding of miR-544a to its target gene. RESULTS: Using mice models with spinal cord injury, we found that the strength of their four limbs began to recover 7 days after injury. The results of microarray and qRT-PCR confirmed that mir-544a level in mice with spinal cord injury decreased with increase of injury time, while the levels of inflammatory genes MCP-1 (monocyte chemoattractant protein-1), IL1 (interleukin-1) and TNF-α (tumor necrosis factor alpha) IL36B (interleukin-36 beta) and IL17B (interleukin-17 beta) were significantly increased. However, overexpression of miR-544a in the mice significantly reduced the level of inflammation and restored their grip strength in their four limbs. Finally, we found that miR-544a can bind to the NEUROD4 (Neurogenic differentiation 4) 3'UTR (Untranslated Region) region through bioinformatics website prediction, which was further confirmed by dual luciferase reporter assay. NEUROD4 level was significantly reduced following the overexpression of miR-544a. CONCLUSION: The expression of miR-544a was significantly decreased after spinal cord injury. High expression of miR-544a could alleviate the inflammation caused by spinal cord injury and promote the recovery of spinal cord via the inhibition of NEUROD4.

FOXP4-AS1 participates in the development and progression of osteosarcoma by downregulating LATS1 via binding to LSD1 and EZH2.

Osteosarcoma (OS) is the most common malignant bone tumor in children and adolescents. LncRNA has been confirmed to participate in a variety of cancers. The purpose of this study was to explore the effect of FOXP4-AS1 on the development of osteosarcoma (OS) and its underlying mechanism. FOXP4-AS1 expressions in 60 OS tissues and paracancerous tissues were detected by qRT-PCR (quantitative real-time polymerase chain reaction). We confirmed that FOXP4-AS1 was overexpressed in OS tissues than that of paracancerous tissues. The disease-free survival and overall survival of OS patients were not correlated with age, gender and tumor location, but remarkably correlated with FOXP4-AS1 expression, tumor size and lung metastasis. For in vitro experiments, MG63 cells expressed a higher expression of FOXP4-AS1, whereas U2OS cells expressed a lower expression, which were selected for the following studies. Overexpressed FOXP4-AS1 led to enhanced proliferation, migration and invasion, shortened G0/G1 phase, as well as inhibited cell cycle. Knockdown of FOXP4-AS1 in MG63 cells obtained the opposite results. Furthermore, RIP assay indicated that FOXP4-AS1 could inhibit LATS1 expression by binding to LSD1 and EZH2, so as to participate in OS development. In conclusion, these results revealed that FOXP4-AS1 is overexpressed in OS, and is the independent risk factor in OS prognosis. Upregulated FOXP4-AS1 promotes the proliferation, migration and cell cycle, but inhibits apoptosis of OS cells. Furthermore, FOXP4-AS1 participates in the development and progression of OS by downregulating LATS1 via binding to LSD1 and EZH2.

The Role of Minimally Invasive Vertebral Body Stent on Reduction of the Deflation Effect After Kyphoplasty: A Biomechanical Study.

STUDY DESIGN: Biomechanical investigation using cadaver spines. OBJECTIVE: The aim of the present study was to assess the magnitude of the deflation effect after balloon kyphoplasty (BKP) or use of minimally invasive vertebral body stent (MIVBS) in in vitro biomechanical condition. SUMMARY OF BACKGROUND DATA: BKP is a well-established minimally invasive treatment option for osteoporotic vertebral compression fractures. However, this technique can lead to a secondary height loss-known as the "deflation effect"-causing intrasegmental kyphosis and an overall alignment failure. METHODS: The study was conducted on 24 human cadaveric vertebral bodies (T12-L5). After creating a compression fracture model, the fractured vertebral bodies were reduced by BKP (n = 12) or by MIVBS (n = 12) and then augmented with polymethyl methacrylate bone cement. Each step of the procedure was performed under fluoroscopic guidance and the results were analyzed quantitatively. Finally, the strength and stiffness of augmented vertebral bodies were measured by biomechanical tests. RESULTS: Complete initial reduction of the fractured vertebral body height was achieved by both systems. Secondary loss of reduction after balloon deflation was significantly greater in the BKP group (2.36 ±â€Š0.63 mm vs. 0.34 ±â€Š0.43 mm in the MIVBS group; P < 0.05). Height gain was significantly higher in the MIVBS group (77.68% ±â€Š11.46% vs. 34.87% ±â€Š13.16% in the BKP group; P < 0.05). Increase in the kyphotic angle gain (relative to the preoperative kyphotic angle) was significantly more in the MIVBS group (95.60% ±â€Š6.12% vs. 77.0% ±â€Š4.94% in the BKP group; P < 0.05). Failure load was significantly higher in the MIVBS group (189% ±â€Š16% vs. 146% ±â€Š14%; P < 0.05). However, stiffness was not significantly different between the two groups. CONCLUSION: The deflation effect after BKP can be significantly decreased with the use of the MIVBS technique. LEVEL OF EVIDENCE: N/A.

Application of 3D printing rapid prototyping-assisted percutaneous fixation in the treatment of intertrochanteric fracture.

The aim of the present study was to investigate the application of 3D printing (3DP) rapid prototyping (RP) technique-assisted percutaneous fixation in the treatment of femoral intertrochanteric fracture (ITF) using proximal femoral nail anti-rotation (PFNA). A total of 39 patients with unstable ITF were included in the current study. Patients were divided into two groups: 19 patients were examined using computed tomography scanning and underwent PFNA with SDP-RP whereas the other 20 patients underwent conventional PFNA treatment. Anatomical data were converted from the Digital Imaging and Communications in Medicine format to the stereolithography format using M3D software. The 3DP-RP model was established using the fused deposition modeling technique and the length and diameter of the main screw blade was measured during the simulation. The postoperative femoral neck-shaft angle (NSA), surgery duration, intraoperative and postoperative blood loss, and the duration of hospital stay were recorded and compared with the corresponding values in conventional surgery. No significant differences were observed in mean PFNA size between the implants used and the preoperative planning estimates. It was demonstrated that the 3DP-RP assisted procedure resulted in more effective reduction of the NSA. Furthermore, patients undergoing 3DP-RP experienced a significant reduction in duration of surgery (P<0.01), as well as reductions in intraoperative (P=0.02) and postoperative (P=0.03) blood loss, compared with conventional surgery. At 6 months post-surgery, no cases of hip varus/vague deformities or implant failure were observed in patients that underwent either the 3DP-RP-assisted or conventional procedure. The results of the present study suggest that the 3DP-RP technique is able to create an accurate model of the ITF, which facilitates surgical planning and fracture reduction, thus improving the efficiency of PFNA surgery for ITFs.

Analysis of the relationship between the facet fluid sign and lumbar spine motion of degenerative spondylolytic segment using Kinematic MRI.

PURPOSE: To investigate the relationship between the facet fluid sign (FFS) and kinematics of affected lumbar segment in patients with low-grade, single-level lumbar degenerative spondylolisthesis using kinematic magnetic resonance imaging (kMRI).Materials and methods Ninety-two patients diagnosed with low-grade (Grade 1 or 2), single level lumbar degenerative spondylolisthesis L3 to S1. Angular and translational motion, whole lumbar motion, slip percentage, and facet angle were measured. Disc degeneration and degrees of osteoarthritis were graded, facet effusion was evaluated and classified as Positive or Negative FFS (PosFFS or NegFFS).Results There were significant differences in the facet degeneration, translational motion, percentage of total angular motion, and flexion-extension slip difference between the PosFFS and NegFFS group. 28.3% of the patients in PosFFS group had anterolisthesis in flexion, but not extension.There was a significant difference in the incidence of FFS between the dynamic and static spondylolisthesis (75% vs 35%, p < 0.001). If FFS was present on kMRI, a positive predictive value for dynamic spondylolisthesis was 75.6%. with the sensitivity and specificity of 0.75 and 0.65 for posFFS to diagnose dynamic DS. The likelihood ratio for dynamic slips in the presence of FFS was 2.1, and the likelihood ratio for static slips in the presence of FFS was 1.1.Conclusion Facet fluid sign was present in weight-bearing neutral position, and the presence of FFS in kMRI increased the probability of having dynamic spondylolisthesis.

Prognosis for recipients with hepatocellular carcinoma of salvage liver transplantation versus those of primary liver transplantation: a retrospective single-center study.

PURPOSE: The prognosis for recipients with hepatocellular carcinoma (HCC) of salvage liver transplantation (SLT) versus those of primary liver transplantation (PLT) remains controversial. The objective of this study was to evaluate the clinical features and survival rate of SLT recipients. METHODS: Three hundred seventy-one patients with HCC transplanted at Shanghai General Hospital, China, between October 2001 and October 2011 were separated into PLT (n = 295) and SLT (n = 76) groups. Patient characteristics and survival curves were studied by univariate and multivariate analysis. A Milan criteria-stratified survival analysis was conducted. RESULTS: The proportions of reoperation (11.8 vs. 5.4 %, P = 0.047) and early postoperative mortality (11.8 vs. 4.7 %, P = 0.032) were higher in the SLT group than in the PLT group. Recurrence free survival (RFS) rate and overall survival (OS) rate had no statistically significant differences after stratification using Milan criteria between the PLT group and SLT group. Alphafetoprotein >400 ng/mL (P = 0.011), microscopic vascular invasion (MVI) (P < 0.001), tumor node metastasis (TNM) staging (P = 0.006), and out of Milan criteria (P < 0.001) were independent risk factors for RFS, while MVI (P < 0.001), TNM staging (P = 0.009), and out of Milan criteria (P = 0.003) were factors for OS. In the multivariate logistic regression analysis, HCC recurrence was associated with MVI (OR = 4.196 [2.538-6.936], P < 0.001), and out of Milan criteria (OR = 2.704 [1.643-4.451], P < 0.001). CONCLUSIONS: Our retrospective, single-center study demonstrated that SLT increases surgical difficulty; however, it has good post-transplantation OS and is a feasible alternative after HCC recurrence within Milan criteria.

The effects of a semiconstrained integrated artificial disc on zygapophyseal joint pressure and displacement.

STUDY DESIGN: Measurement of zygapophyseal joint pressure and displacement was performed after placement of a semiconstrained integrated artificial disc (SIAD) in a cadaver model. OBJECTIVE: To understand the likelihood of accelerated zygapophyseal joints degeneration as a result of the implant. SUMMARY OF BACKGROUND DATA: A SIAD has been developed to treat lumbar spondylosis secondary to segmental disc degeneration and spinal stenosis. The SIAD replaces the stenotic segment's disc. Previous studies have demonstrated that nonconstrained artificial disc (NAD) replacements fail to achieve their optimal long-term outcomes likely because of significantly increased zygapophyseal joint pressure and displacement at the implanted level. Moreover, clinical studies have reported an increased incidence of zygapophyseal joint degeneration after lumbar disc replacement. METHODS: Eight cadaver lumbar specimens (L2-L5) were loaded in flexion, neutral, extension, left bend, and right rotation. Zygapophyseal joint pressure and displacement were measured during each of the 5 positions at each of the 3 levels with the ratio of deformation calculated under the different loads. An artificial disc was placed at the L3-L4 level, and the measurements were repeated. RESULTS: After L3-L4 segment implantation, the pressure in the zygapophyseal joint at operative segment was not significantly changed by SIAD and NAD implantation in axial compression and flexion, compared with physiological disc. Notable differences in zygapophyseal joint pressure between the SIAD and NAD were identified at the operative level in extension, left bend, and right rotation. The adjacent-level effect of NAD was significantly greater than that of SIAD. The ratio of deformation difference between the 2 discs was increased by load experienced in extension, flexion, left bend, and right rotation. CONCLUSION: The SIAD provided a superior biomechanical milieu for zygapophyseal joints at the implanted and adjacent levels compared with NAD, which may avoid the acceleration of postoperative zygapophyseal joint degeneration. LEVEL OF EVIDENCE: 1.

Biomechanical effects of semi-constrained integrated artificial discs on zygapophysial joints of implanted lumbar segments.

This study aimed to optimize the design and application of semi-constrained integrated artificial discs (SIADs) using a finite element (FE) analysis following implantation, wherein the zygapophysial joints of the segment were biomechanically reconstructed. An FE model of the L4-L5 segment was constructed. Variations in the stresses on the discs and zygapophysial joints were observed during 5° anteflexion, 5° extension and 5° rotation under the 400-N applied axial load. Stresses and load translation analyses of the discs and zygapophysial joints were conducted during anteflexion, extension and rotation under the 400-N applied axial load. Following implantation of the lumbar segments, the stresses on the SIAD zygapophysial joints were not significantly different from those of physiological discs during anteflexion, and these were both marginally greater compared with those of non-constrained artificial discs (NADs). During extension, the increase in the stress on the SIAD zygapophysial joints was less than that on NAD zygapophysial joints. Stresses on the NAD zygapophysial joints were higher than those on SIAD and physiological discs during rotation. The stress on the SIAD zygapophysial joints was not significantly different from that on physiological discs during rotation. For SIADs and NADs, the stresses on the zygapophysial joints and the displacements of the discs were greater compared with those of the physiological discs during extension. The SIADs affected the variations in the stresses on the implanted segment more than the NADs, and the SIADs protected the zygapophysial joints of the implanted segment to a higher degree than the NADs.