Harmine loaded Au@MSNs@PEG@Asp6 nano-composites for treatment of spinal metastasis from lung adenocarcinoma by targeting ANXA9 in vivo experiment.

Background: Annexin A9 (ANXA9) has been proved to be concerned with cancer development. However, to explore the clinical consequences of ANXA9 in lung adenocarcinoma (LUAD), especially its correlation to spinal metastasis (SM) has no in-depth study. The study was expected to elucidate the mechanism of ANXA9 in regulating SM of LUAD and create a productive nano-composites delivery system targeting this gene for treatment of SM. Methods: Harmine (HM), a ß-carboline extracted from the traditional Chinese herb Peganum harmala, loaded Au@MSNs@PEG@Asp6 (NPS) nano-composites were synthesized. Bioinformatics analysis and clinical specimens' tests were used to verify the association between ANXA9 and prognosis of LUAD with SM. The immunohistochemistry (IHC) was employed to detect the expression levels of the ANXA9 protein in LUAD tissues with or without SM, and its significance in clinic was also explored. ANXA9­siRNA was applied to investigate the molecular mechanism of ANXA9 in tumor behaviors. The HM release kinetics was detected by high performance liquid chromatography (HPLC) method. The cellular uptake efficiency of nanoparticles by A549 cells was observed by fluorescence microscope. Antitumor effects of nanoparticles were assessed in the nude mouse model of SM. Results: The genomic amplification of ANXA9 was prevalent in LUAD tissues and closely associated with poor outcome and SM (P<0.01). The experimental result manifested that high expression of ANXA9 could lead to wretched prognosis and ANXA9 was an independent risk factor for survival (P<0.05). After impeding expression of ANXA9, the proliferation and metastatic ability of tumor cells obviously decreased, and expression of matrix metallopeptidase 2 (MMP-2) and matrix metallopeptidase 9 (MMP-9) were considerably downregulated, while the expression of associated oncogene pathway were downregulated (P<0.01) as well. The synthesized HM-loaded NPS nano-composites could target to cancer and response to reactive oxygen species (ROS) to release HM slowly. Notably, in comparison to free HM, the nano-composites showed excellent targeting and anti-tumor effects in the A549 cell-bearing mouse model. Conclusions: ANXA9 may serve as a novel biomarker for predicting poor prognosis in LUAD, and we provided an efficient and targeting drug delivery nano-composites system for precise treatment of SM from LUAD.

Genome-wide analysis of long non-coding RNA expression profile in lung adenocarcinoma compared to spinal metastasis.

BACKGROUND: Long non-coding RNAs (lncRNAs) play important roles in tumor metastasis. The aim of the present study was to investigate their expression profile and potential functions in spinal metastasis (SM) of lung adenocarcinoma. METHODS: We conducted lncRNA and mRNA expression in lung adenocarcinoma and its SM tissue using microarray analysis. Quantitative reverse-transcriptase polymerase chain reaction (qRT-PCR) revealed 10 differentially expressed lncRNAs. Gene ontology and pathway analysis were performed to test the gene effect. Possible target genes of lncRNAs were predicted based on precise algorithms. RESULTS: Microarray analysis found many significantly differentially expressed lncRNAs and mRNAs in lung adenocarcinoma compared with SM. qRT-PCR results aligned with those of the microarray analysis. The expression level of 10 lncRNAs showed the same trend (P<0.05). Biologic pathways known to be involved in cancer were identified among the differentially expressed mRNAs; these include cell adhesion molecules (related to 42 genes), focal adhesion (related to 31 genes), cytokine-cytokine receptor interaction (related to 48 genes), and extracellular matrix-receptor interaction (related to 23 genes). About 9,458 lncRNAs were found to have cis- or trans-genes. A total of 2,317 cis target genes were discovered to be abnormally expressed and could be regulated by lncRNAs in SM of lung adenocarcinoma. CONCLUSIONS: Our results offer a genome-wide differential expression of lncRNA in lung adenocarcinoma and SM, as well as laying the foundation for further investigations of lncRNAs correlated with lung adenocarcinoma metastasis.

Diagnostic accuracy of MR, CT, and ECT in the differentiation of neoplastic from nonneoplastic spine lesions.

AIM: To provide guidance for appropriate imaging examinations for diagnosing spinal tumors or tumor-like lesions. METHODS: A total of 121 patients with suspected spinal tumors were included this retrospective study. Each patient underwent ≥2 imaging examinations, including computerized tomography (CT), magnetic resonance (MR), and/or emission computed tomography (ECT). All patients were diagnosed by pathology after core needle or surgical biopsies. The results were compared with those of pathological examinations using paired chi-squared tests, and compared with each other. Statistical indicators that tested the consistency of the results included McNemar's and kappa coefficients, as well as receiver operating characteristic curves. RESULTS: The differences among MR, CT, ECT, and pathology were not significant. The kappa coefficient of MR, CT, and ECT was 46.1%, 36.0%, and 55.9%, respectively. The area under the curve of ECT, MR, and CT scans was 0.809, 0.705, and 0.704, respectively; and the differences among them were significant (P < .05). Post hoc multiple comparisons showed no significant differences among imaging examinations in terms of sensitivity, specificity, misdiagnosis rate, and coincidence rate (P > .05). However, significant differences were noted in the kappa coefficient and missed diagnosis rate (P < .05). CONCLUSIONS: Although ECT was the most accurate imaging method, its high cost and large radiation dosage limit its widespread application. Furthermore, MR verified spinal tumors more effectively; however, CT excluded them more efficiently. In summary, when all factors are considered, MR is still the optimal modality for the diagnosis of spinal tumors, especially during the initial screening.

CX3CL1 Induces Vertebral Microvascular Barrier Dysfunction via the Src/P115-RhoGEF/ROCK Signaling Pathway.

Trans-endothelial migration (TEM) of cancer cells is a critical step in metastasis. Micro-vascular barrier disruptions of distant organs play important roles in tumor cells TEM. The spine is a preferred site for multiple cancer cell metastases. Our previous study found that vertebral spongy bone was rich in CX3CL1 and that CX3CL1 can attract fractalkine receptor-expressing tumor cells to the spine. In the present study, we determined whether CX3CL1 was involved in vertebral micro-vascular barrier disruption and promoted tumor cell TEM after circulating tumor cells were arrested in the vertebral micro-vasculature. We examined the role of CX3CL1 in the barrier function of vertebral micro-vascular endothelial cells (VMECs) and explored the molecular mechanisms of CX3CL1-induced VMEC barrier disruption. Our results demonstrated that CX3CL1 led to F-actin formation and ZO-1 disruption in VMECs and induced the vertebral micro-vascular barrier disruption. Importantly, we found that the activation of the Src/P115-RhoGEF/ROCK signaling pathway plays an important role in CX3CL1-induced VMEC stress fiber formation, ZO-1 disruption and then vertebral micro-vascular barrier hyper-permeability. Inhibiting Src/P115-RhoGEF/ROCK signaling in VMECs effectively blocked CX3CL1-induced vertebral vascular endothelial dysfunction and subsequent tumor cell TEM. The results of this study and our previous study indicate that in addition to its chemotaxis, CX3CL1 plays a critical role in regulating vertebral micro-vascular barrier function and tumor cell TEM. CX3CL1 induced VMECs stress fiber formation, ZO-1 disruption and then vascular endothelial hyperpermeability via activation of the Src/P115-RhoGEF/ROCK signaling pathway. The inhibition of the Src/P115-RhoGEF/ROCK signaling pathway in VMECs effectively blocked tumor cells TEMs in vertebral spongy bone and maybe a potential therapeutic strategy for spine metastases in the future.

Relationships of accelerometer-based measured objective physical activity and sedentary behaviour with cognitive function: a comparative cross-sectional study of China's elderly population.

BACKGROUND: This study explored the effects of physical activity and sedentary behaviour on the decline of cognitive ability among the elderly. To compensate for the limitations of self-reported physical activity, objective measures were used. METHODS: A cross-sectional survey of 308 aged people mean 68.66 ± 5.377 years, in Nanjing, China, was conducted. Physical activity was measured using the ActiGraph GT3X+, and cognitive function was measured using the Montreal Cognitive Assessment. RESULTS: The overall participant model, adjusted for age, BMI, education, and monthly average income, found that light physical activity (ß = 0.006, p < 0.01), moderate-vigorous physical activity (ß = 0.068, p < 0.001), and total physical activity (ß = 0.006, p < 0.01) had a significant linear relationship with cognitive ability, while sedentary time did not (ß = - 0.020, p>0.05). Further, light physical activity only affects the cognitive ability of elderly females (ß = 0.006, p < 0.05). There was an inverted 'U' association between moderate-vigorous physical activity and cognitive ability. The association models found that moderate-vigorous physical activity in the 22.13 min·day- 1~38.79 min·day- 1 range affected cognitive ability most beneficially, with the highest beta coefficient among all groups (ß = 0.091, p < 0.05). CONCLUSIONS: While physical activity can significantly improve cognitive ability among the elderly, sedentary behaviour is associated with decreased cognitive function across genders.

ADAM17-regulated CX3CL1 expression produced by bone marrow endothelial cells promotes spinal metastasis from hepatocellular carcinoma.

Spinal metastasis occurs in 50­75% of bone metastases caused by hepatocellular carcinoma (HCC), and HCC­derived spinal metastasis can lead to a less favorable prognosis. Recently, several studies have demonstrated that C­X3­C motif chemokine ligand 1 (CX3CL1) is closely associated with cancer metastasis, and its secretion is modulated by a disintegrin and metalloproteinase 17 (ADAM17). Bone marrow endothelial cells (BMECs) are an essential component of bone marrow. However, little is known about the roles in and effects of BMECs on HCC spinal metastasis. The present study demonstrated that CX3CL1 and C­X­C motif chemokine receptor 3 (CXCR3) expression was upregulated in HCC spinal metastases, and that CX3CL1 promoted the migration and invasion of HCC cells to the spine. Western blot analysis revealed that the Src/protein tyrosine kinase 2 (PTK2) axis participated in CX3CL1­induced HCC cell invasion and migration. CX3CL1 also increased the expression of M2 macrophage markers in THP­1 monocytes. BMECs promoted the migration and invasion of Hep3B and MHCC97H cells by secreting soluble CX3CL1, whereas the neutralization of CX3CL1 inhibited this enhancement. CX3CL1 enhanced the activation of the phosphatidylinositol­4,5­bisphosphate 3­kinase catalytic subunit alpha (PIK3CA)/AKT serine/threonine kinase 1 (AKT1) and Ras homolog family member A (RHOA)/Rho associated coiled­coil containing protein kinase 2 (ROCK2) signaling pathways through the Src/PTK2 signaling pathway. Furthermore, ADAM17 was activated by mitogen­activated protein kinase (MAPK)z14 in BMECs and significantly promoted the secretion of CX3CL1. HCC cells enhanced the recruitment and proliferation of BMECs. The overexpression of CX3CR1 facilitated the spinal metastasis of HCC in a mouse model in vivo. In addition, in vivo experiments revealed that BMECs promoted the growth of HCC in the spine. The present study demonstrated that CX3CL1 participates in HCC spinal metastasis, and that BMECs play an important role in the regulation of CX3CL1 in the spinal metastatic environment.

Thermosensitive hydrogels loaded with human-induced pluripotent stem cells overexpressing growth differentiation factor-5 ameliorate intervertebral disc degeneration in rats.

To evaluate the effects of thermosensitive hydrogels loaded with human-induced pluripotent stem cells transfected with the growth differentiation factor-5 (GDF5-hiPSCs) on rat intervertebral disc regeneration. GDF5-hiPSCs were cocultured with rat nucleus pulposus (NP) cells in vitro. Real-time PCR and western blot were used to determine the differentiation of hiPSCs. Rat caudal intervertebral discs were punctured using a needle under X-ray, and groups of coccygeal (Co) discs were subject to various treatments: Puncture group (Co6/7, punctured without treatment); Hydrogel group (Co7/8, 2 µl of hydrogel injected without cells); GDF5-hiPSCs + Hydrogel group (Co8/9, 2 µl of GDF5-hiPSCs-loaded hydrogel injected); and Normal control (Co5/6). X-ray, MRI, and histological evaluations were performed at 1, 2, and 3 months after cell transplantation and relative changes in the disc height index (DHI%) and voxel count were calculated and compared. GDF5-hiPSCs were successfully differentiated to a chondrogenic linage after cocultured with rat NP cells. In terms of X-ray, MRI, and HE staining scores, the GDF5-hiPSCs + Hydrogel group was significantly superior to the Puncture and Hydrogel groups (p < .05). Compared with the Normal group, the MRI-based voxel count of the GDF5-hiPSCs + Hydrogel group was significantly lower at 1, 2, and 3 months after cell transplantation (p < .05). However, there were no significant differences in histological scores at 1 and 2 months after cell transplantation compared with the Normal group (p > .05). In conclusion, thermosensitive hydrogel-encapsulated hiPSCs overexpressing the GDF5 gene ameliorated intervertebral disc degeneration.

Hsa_circ_0006571 promotes spinal metastasis through sponging microRNA-138 to regulate sirtuin 1 expression in lung adenocarcinoma.

BACKGROUND: Circular RNAs (circRNAs) are known to participate in lung cancer. However, their role in spinal metastasis (SM) of lung adenocarcinoma remains elusive. In this study, we determined that hsa_circ_0006571 serves as a sponge for miR-138, which targets sirtuin 1 (Sirt1) in the development of SM. METHODS: A human circRNA microarray was performed to compare SM and lung adenocarcinoma samples. The expression of hsa_circ_0006571 and miR-138 was determined using quantitative polymerase chain reaction (qPCR) in vitro and in vivo. Cell proliferation was performed by Cell Counting Kit-8 (CCK-8) and apoptosis was analyzed by Annexin V/PI staining. RNA-pulldown and RNA immunoprecipitation (RIP) were used to analyze the interaction between hsa_circ_0006571. Tumor metastasis was determined through a xenograft experiment in vivo. RESULTS: Hsa_circ_0006571 was observed to be significantly upregulated in SM tissues through circRNA microarray and qPCR. We detected a lower expression of miR-138 in SM tissues compared with lung adenocarcinoma. Hsa_circ_0006571 silencing suppressed lung cancer cell proliferation and migration while promoting apoptosis. Hsa_circ_0006571 interacted with miR-138 to promote expression of Sirt1, leading to activation of epithelial-mesenchymal transition (EMT). Xenograft experiments showed that downregulation of hsa_circ_0006571 delayed the SM of lung adenocarcinoma cells via the miR-138-Sirt1 axis. CONCLUSIONS: Hsa_circ_0006571 promoted tumor cell migration and invasion via the miR-138/Sirt1 pathway. Our observations indicate that circRNAs are possible novel therapeutic targets for SM of lung adenocarcinoma.

Influence of the built environment of Nanjing's Urban Community on the leisure physical activity of the elderly: an empirical study.

BACKGROUND: Urbanization and aging are global phenomena that offer unique challenges in different countries. A supportive environment plays an important role in addressing the issues of health behavioral change and health promotion (e.g., prevent chronic illnesses, promote mental health) among older adults. With the development of the socio-ecological theoretical model, studies on the impact of supportive environments on physical activity have become popular in the public health field in the EU and US. Meanwhile, very few Chinese studies have examined the relationship between built environment features and older adults' physical activity at the ecological level. The purpose of the study is to investigate how the factors part of the built environment of Nanjing's communities also influence leisure time physical activity among the elderly. METHODS: Using a socio-ecological model as a theoretical framework, we conducted a cross-sectional study of 399 elderly people from 19 communities in Nanjing, China, using a one-on-one questionnaire to collect data, including participants' perceived built environment and self-reported physical activity. A multivariate linear regression method was used to analyze the factors influencing their recreational physical activity. RESULTS: This study found that compared to older people with low average monthly income, the recreational physical activity of the elderly with average monthly incomes between 1001 and 2000 ¥ (ß = 23.31, p < 0.001) and 2001 ¥ or more (ß = 21.15, p < 0.001) are significantly higher. After controlling for individual covariates, street connectivity (ß = 7.34, p = 0.030) and street pavement slope (ß = - 7.72, p = 0.020), we found that two out of ten built environment factors indicators influence their physical activity. The importance of each influencing factor ranked from highest to lowest are monthly average income, street pavement slope, and street connectivity. Other factors were not significantly related to recreational physical activity by the elderly. CONCLUSIONS: Older adults with a high income were more likely to participate in recreational physical activity than those with a low income. In order to positively impact physical activity in older adults and ultimately improve health, policymakers and urban planners need to ensure that street connectivity and street pavement slope are factored into the design and development of the urban environment.

Multi-segmental lumbar spinal stenosis treated with Dynesys stabilization versus lumbar fusion in elderly patients: a retrospective study with a minimum of 5 years' follow-up.

INTRODUCTION: Middle- and long-term outcomes of multi-segmental lumbar spinal stenosis treated with Dynesys stabilization (DS) have rarely been reported. Older age and multi-segmental degeneration may be positive factors in achieving satisfactory outcomes following DS. The present study aimed to compare the middle- and long-term outcomes of DS with lumbar fusion for treatment of multi-segmental lumbar spinal stenosis (ms-LSS) in elderly patients. MATERIALS AND METHODS: This study retrospectively analyzed patients with ms-LSS treated by DS or lumbar fusion from January 2011 to April 2013. Twenty-two patients were included in the Dynesys group, and 44 patients treated by lumbar fusion and rigid fixation were included in the fusion group. Clinical outcomes were assessed by VAS and ODI. Radiological outcomes were measured by range of motion (ROM) of stabilized segments and the proximal adjacent segment, intervertebral disc height (DH) and L1-S1 lumbar lordosis angle (LL). Modified Pfirrmann grade score was used to access disc degeneration. OUTCOMES: The mean follow-up time of the Dynesys group and fusion group was 68.50 ± 6.40 and 70.14 ± 7.26 months, respectively. Baseline data were similar between the two groups. There were no significant differences between the two groups in terms of improvement of clinical outcomes (VAS and ODI). DS preserved a certain degree of ROM (3.74 ± 2.00) of surgical segments. ROM of proximal adjacent segment underwent an increase in both groups at the final follow-up. The DH of the surgical segments and proximal adjacent segment in both groups was significantly lower than that before surgery (P = 0.000). LL of both groups improved (P = 0.000), and there was no significant difference between the two groups. The modified Pfirrmann score of proximal adjacent segment of both groups increased at the final follow-up. The fusion group underwent a more significant increase (P = 0.000), whereas the inter-group difference showed no significance (P = 0.090). CONCLUSION: DS is a safe and effective surgical treatment of multi-segmental lumbar spinal stenosis in the elderly population. DS preserves a certain degree of mobility of surgical segments.

ß­catenin signalling inhibits cartilage endplate chondrocyte homeostasis in vitro.

Cartilaginous endplate degeneration serves a key role in the process of intervertebral disc (IVD) degeneration, however, effective therapies are hindered by an incomplete understanding of the mechanisms that underlie cartilage endplate (CEP) homeostasis and degeneration. Wnt/ß­catenin signalling has been reported as a major factor in regulating biological processes. Whether Wnt/ß­catenin signalling engages in CEP homeostasis has not yet been investigated. The present study aimed to assess the function of CEP cells via the activation of Wnt/ß­catenin signalling to examine and promote the mechanism of degeneration of CEP in vitro. Rat CEP cells were confirmed to exhibit a chondrocytic phenotype by toluidine blue staining. The increased number of senescence­associated ß­galactosidase (SA­ß­gal)­positive cells and reduced cellular proliferation were investigated in the presence of a ß­catenin inhibitor, and the inhibitor improved the trend of senescence. An increased number of apoptotic cells was detected by lithium chloride treatment, and inhibiting Wnt/ß­catenin signalling protected the cells from apoptosis. Expression of the catabolic enzymes, metalloproteinase­13 and a disintegrin and metalloproteinase with thrombospondin motifs­5, and the decreased expression of aggrecan were also observed by Wnt/ß­catenin signalling activation, and a Wnt/ß­catenin signalling inhibitor decreased the expression of catabolic enzymes and increased the expression of aggrecan induced by Wnt/ß­catenin signalling activation. Wnt/ß­catenin signalling may provide potential strategies for preventing CEP degeneration.

Value of CT-guided Core Needle Biopsy in Diagnosing Spinal Lesions: A Comparison Study.

OBJECTIVE: A retrospective study was designed to evaluate the effectiveness of CT-guided core needle biopsy in diagnosing spinal lesions through comparison with C-arm guidance. METHODS: From April 2013 to July 2017, a total of 188 patients, who suffered from spinal lesions or had malignant tumor history with a new spinal fracture, were included in this study. There were 96 men and 92 women, with an average of 57.1 years. A total of 238 core needle biopsies were performed. A total of 140 core needle biopsies were carried out under C-arm guidance in 102 patients (group 1); 98 core needle biopsies were carried out under CT guidance in 86 patients (group 2); 108 core needle biopsies were performed in thoracic vertebrae, 116 were in lumbar vertebrae, and 14 were in sacral vertebrae. Seventy-eight patients accepted surgical treatment after biopsies. For these patients, the histological pathologies of the biopsy and surgery were compared to evaluate the accuracy of the biopsy. For the other 110 patients who did not receive surgical treatment, the treatment response and the clinical course were used to evaluate the accuracy of the biopsy. The success rate, the diagnostic accuracy rate, the true positive/negative rate, and complications of the two groups were calculated and compared. RESULTS: There were no significant differences in sex, age, and lesion sites between the C-arm guidance group (group 1) and the CT guidance group (group 2). There were no complications in the two groups. Pathological diagnoses were established in 232 of 238 biopsies. They revealed that 52 were primary malignant tumors, 12 were benign tumors, 70 were metastatic tumors, 4 were tuberculosis, and 94 were classified as "other." The success rate of group 2 was higher than that of group 1, but it was not statistically significant (95.7% vs 100%; P = 0.098). According to the final diagnosis, the diagnostic accuracy rates were calculated and compared. There was no significant difference between the two groups (95.5% vs 96.9%; P = 0.835). The kappa coefficient was used to analyze the concordance between the histological pathologies of the biopsy and the final diagnosis in the two groups. The kappa values of the two group were 0.909 and 0.939, respectively. The results showed good consistency in both groups, but seemed better for group 2. CONCLUSION: CT-guided core needle biopsy is a relatively safe and effective procedure for diagnosing spinal lesions with a high diagnostic accuracy rate and few complications.

Surgical Outcomes and Prognostic Factors for Metastatic Spine Hepatocellular Carcinoma.

BACKGROUND: We evaluated the surgical outcomes of metastatic spine hepatocellular carcinoma (HCC) and determined the factors that might influence the outcomes of metastatic HCC of the spine. METHODS: From 2010 to 2017, 72 patients with HCC-derived metastatic spine tumors were treated in our department. For each patient, we recorded the pre- and postoperative visual analog scale score, Frankel grade, perioperative complications, and mortality. Univariate and multivariate analyses were used to explore a range of factors that might influence postoperative survival. RESULTS: The mean postoperative survival was 10.8 ± 5.4 months. The concordance rate between a Tokuhashi score of 0-8 and a life expectancy of <6 months was only 19.2%. The mean postoperative survival for patients undergoing excisional surgery was 14.7 ± 6.5 months, and the mean survival of those receiving palliative surgery was 8.5 ± 2.6 months. Pain had significantly improved in both patient groups (P < 0.001). Paralysis did not change significantly in the excisional surgery group (P = 0.641) or palliative surgery group (P = 0.912). Univariate analysis showed that the preoperative Frankel score, Tomita score, Tokuhashi score, blood loss, multilevel metastases, and operative type were independent prognostic factors for postoperative survival time. Multivariate analysis showed that operation type was an independent factor for prognosis, just as were the Tomita score and Tokuhashi score. CONCLUSIONS: Our results have challenged previously reported estimates of the life expectancy correlating with the Tokuhashi score. Our results showed that excisional surgery resulted in better clinical outcomes compared with palliative surgery.

LINC00852 Promotes Lung Adenocarcinoma Spinal Metastasis by Targeting S100A9.

Background: Lung adenocarcinoma has a strong tendency to develop into bone metastases, especially spinal metastases (SM). Long noncoding RNAs (lncRNAs) play critical roles in regulating several biological processes in cancer cells. However, the mechanisms underlying the roles of lncRNAs in the development of SM have not been elucidated to date. Methods: Clinical specimens were collected for analysis of differentially expressed lncRNAs. The Kyoto Encyclopedia of Genes and Genomes (KEGG) was used to examine the effects of these genes on pathways. RNA pull-down was utilized to identify the targeting protein of lncRNAs. The effects of lncRNA on its target were detected in A549 and SPCA-1 cells via perturbation of the lncRNA expression. Oncological behavioral changes in transfected cells and phosphorylation of kinases in the relevant pathways, with or without inhibitors, were observed. Further, tumorigenicity was found to occur in experimental nude mice. Results: LINC00852 and the mitogen-activated protein kinase (MAPK) pathway were found to be associated with SM. Moreover, the LINC00852 target S100A9 had a positive regulatory role in the progression, migration, invasion, and metastasis of lung adenocarcinoma cells, both in vitro and in vivo. Furthermore, S100A9 strongly activated the P38 and REK1/2 kinases, and slightly activated the phosphorylation of the JNK kinase in the MAPK pathway in A549 and SPCA-1 cells. Conclusion: LINC00852 targets S100A9 to promote progression and oncogenic ability in lung adenocarcinoma SM through activation of the MAPK pathway. These findings suggest a potential novel target for early intervention against SM in lung cancer.

CX3CL1 involves in breast cancer metastasizing to the spine via the Src/FAK signaling pathway.

C-X3-C chemokine ligand 1 (CX3CL1) has been shown to be involved in the development of multiple tumors. Our previous study demonstrated that CX3CL1 may be involved in the process of metastasis of various malignant tumors to the spine, including breast cancer, but the molecular mechanism was still unknown. In the present study, we found that the receptor CX3CR1 was overexpressed in the spinal metastases of breast cancer than in para-tumor tissue. In terms of CX3CL1, it was significantly more expressed in normal spinal cancellous bone than in limbs. However, CX3CR1 was not expressed at a high level in every breast cancer cell compared with the human mammary epithelial cell line MCF-10A. In addition, CX3CL1 did promote the migration and invasion abilities of MDA-MB-231 cells. However, CX3CL1 has no obvious effect on cell growth. Furthermore, CX3CL1 induced chemotaxis of tumor cells via the Src/FAK signaling pathway. The migration index enhanced by CX3CL1 was dramatically declined using Bosutinib and PF-00562271, which are the inhibitors of Src and FAK signaling pathways, respectively. Therefore, CX3CL1 in spinal cancellous bone attracts CX3CR1-expressing tumor cells to the spine and enhances their migration and invasion abilities through the Src/FAK signaling pathway.

CX3CL1/fractalkine enhances prostate cancer spinal metastasis by activating the Src/FAK pathway.

Chemokines serve important roles in the development of cancer. C-X3-C motif chemokine ligand 1 (CX3CL1) has been demonstrated to promote metastases in different types of tumors. The authors' previous studies demonstrated that the CX3CL1 (also termed fractalkine)/steroid receptor coactivator (Src)/focal adhesion kinase (FAK) signaling pathway is associated with spinal metastasis. In the present study, it was observed that CX3CL1/C-X3-C motif chemokine receptor 1 (CX3CR1) was overexpressed in prostate cancer tissues with spinal metastasis compared with primary tumors. Overexpression of CX3CR1 induced cell proliferation, migration and invasion, and inhibited cellular apoptosis. However, repression of CX3CR1 reduced cell proliferation, migration and invasion, and increased cellular apoptosis. In addition, the Src/FAK pathway was activated by CX3CL1, which depends on the Tyr992 residue of epidermal growth factor receptor (EGFR) for phosphorylation. The inhibitors of these kinases repressed the cell migration induced by CX3CL1 or CX3CR1 overexpression. Furthermore, overexpression of CX3CR1 induced the spinal metastasis of prostate cancer in an in vivo mouse model. Therefore, CX3CL1 and its regulation of the EGFR, Src and FAK pathways may be potential targets for the early prevention of spinal metastasis in prostate cancer.

Are older patients with solitary spinal metastases fit for total en-bloc surgery?

OBJECTIVE: Due to radical resection, total en-bloc spondylectomy (TES) is associated with significant levels of surgical injury and spinal instability, particularly in elderly patients with solitary spinal metastases (SM), whether the possible benefits outweigh the risk requires intense consideration. Our aim was to compare and analyze the impact of age on patient prognosis. PATIENTS AND METHODS: This study investigated TES in 78 consecutive patients with solitary SM, who were divided into Group A (>65 years, n = 32) and group B (<60 years of age, n = 46). Surgical outcomes were assessed according to survival time, local recurrence, neurological function, pain, and quality of life before and after surgery. Differences between groups were statistically compared using analysis of variance (ANOVA) or chi-square tests. RESULTS: There was no significant difference between the two groups in terms of surgery duration, blood loss, blood transfusion or the duration of hospital stay (p > 0.05). Furthermore, there was no significant difference in the median survival time between the two groups (p > 0.05). However, the perioperative complication rate in group A was higher than that in group B (p < 0.05). There was no significant difference in terms of local recurrence rate when compared between group A and group B (p > 0.05), and there were no significant differences in terms of improvements in neurological function, Visual Analogue Scale and Karnofsky scores of patients between the two groups (p > 0.05). CONCLUSION: Older patients can experience survival and local recurrence rates that were similar to those of younger patients. Although older patients are at increased risk of perioperative complications, this factor does not appear to lead to serious adverse outcomes. Older patients are still good candidates to receive TES to cure solitary SM after careful preparation and strict selection.

Microarray analysis of the expression profile of lncRNAs reveals the key role of lncRNA BC088327 as an agonist to heregulin­1ß­induced cell proliferation in peripheral nerve injury.

Heregulin­1ß is capable of promoting the nerve regeneration of acellular nerve allografts with skin­derived precursor differentiated Schwann cell (SC) therapy in peripheral nerve injury. Long non­coding RNAs (lncRNAs) serve important roles in the regulation of gene transcription and trans-lation in multiple biological processes, but its association with the repair of peripheral nerve injury is unexplored. Therefore, in the present study, the aim was to identify novel indicators for peripheral nerve injury, and to detect whether there is an association between lncRNA expression and the treatment effect of heregulin­1ß on this disorder. The expression status of lncRNAs and mRNAs in a well­built rat model with sciatic nerve injury was investigated using microarray assays. Based on the results of the microarray assays and quantitative polymerase chain reaction validation, it was inferred that lncRNA BC088327 was upregulated to the largest extent among all the lncRNAs. According to these findings, the role of BC088327 in peripheral nerve injury was further assessed by detecting the cell viability, cell cycle and apoptosis in a hypoxic SC cell model after suppressing the expression of BC088327 using specific small interfering RNA. Based on the results of the lncRNA microarray assay, 805 lncRNAs were significantly differentially expressed, among which, 323 lncRNAs were upregulated and 482 lncRNAs were downregulated. Based on the results of the mRNA microarray assay, 1,293 lncRNAs were significantly differentially expressed, including 603 upregulated and 690 downregulated lncRNAs. Moreover, knockdown of lncRNA BC088327 suppressed cell viability and induced cell apoptosis and S-phase cell cycle arrest in the SCs. In conclusion, expression profile changes of lncRNAs in peripheral nerve injuries were closely associated with treatment with heregulin­1ß. lncRNA BC088327 may play a synergistic role with heregulin­1ß in repairing peripheral injury, which has the potential be a biomarker for the detection of peripheral injury and a medical target for the development of therapeutic modalities.

Caveolin­1 regulates oxidative stress­induced senescence in nucleus pulposus cells primarily via the p53/p21 signaling pathway in vitro.

Previous studies have indicated that cellular senescence is a critical underlying mechanism of intervertebral disc degeneration. However, the precise mechanism by which cellular senescence accelerates disc degeneration has not been fully elucidated. Caveolin­1 has recently emerged as an important regulator of cellular senescence. Therefore, the aim of the present study was to investigate whether caveolin­1 is involved in nucleus pulposus (NP) cellular senescence during oxidative stress. PCR was used to detect caveolin­1 mRNA expression and protein expression was detected by western blotting. Caveolin­1 expression at the mRNA and protein levels was markedly increased following treatment with tert­butyl hydroperoxide, and an increase in premature senescence was observed, as determined by senescence­associated ß­galactosidase staining and the decline of cellular proliferative ability. In addition, caveolin­1 gene expression was successfully knocked down by lentivirus­mediated RNA interference, which exerted a protective effect against the cellular senescence induced by oxidative stress. Notably, p53 and p21 protein expression, though not p16 protein expression, decreased with caveolin­1 silencing. The results suggested that caveolin­1 may be involved in NP cellular senescence during oxidative stress in vitro, mainly via the p53/p21 signaling pathway. Thus, caveolin­1 may represent a novel therapeutic target for the prevention of intervertebral disc degeneration.

Study of synergistic role of allogenic skin-derived precursor differentiated Schwann cells and heregulin-1ß in nerve regeneration with an acellular nerve allograft.

Development of tissue structure and three-dimensional microenvironment is crucial for regeneration of axons in the peripheral nerve repair. In this study we aimed to evaluate the efficacy of nerve regeneration by using an acellular nerve allograft (ANA) injected with allogenic skin-derived precursor differentiated Schwann cells (SKP-SCs) and heregulin-1ß. Skin-derived precursor cells (SKPs) were generated from dermis of newborn (postnatal day 2) Wistar rats. In a rat model, nerve regeneration was determined across a 15 mm lesion in the sciatic nerve by using an ANA injected with allogenic SKP-SCs and heregulin-1ß. The electrophysiological analysis, histological examination and electron microscopy were involved in this study. Cultured SKPs expressed nestin and fibronectin, and differentiated into cells with phenotype of SCs that presented characteristic markers of p75NGFR and S100-ß. Implantation of SKP-SCs into the rat models by using ANA and allogenic skin-derived precursor differentiated Schwann cells (SKP-SCs) increases sciatic nerve functional index (SFI), peak amplitudes, nerve conduction velocities, number of myelinated fibers within the graft, while reduces incubation period, sciatic nerve injury-induced weight and contractions loss. Using ANA injected with SKP-SCs combined with heregulin-1ß greatly promote peripheral nerve repair in a rat model. Our results suggest that SKP-SCs transplantation with heregulin-1ß represents a powerful therapeutic approach, and facilitates the efficacy of acellular nerve allograft in peripheral nerve injury, though the detailed mechanism remains to be elucidated.