LINC00482 sponged miR-2467-3p to promote bone metastasis of prostate cancer through activating Wnt/ß-catenin signaling pathway.

This study was designed to investigate the biological functions of LINC00482 in prostate cancer (PCa) with bone metastasis. TCGA dataset of PCa was applied for LINC00482 expression analysis and real time PCR was used to verify the expression level of LINC00482 in PCa tissues as well as PCa bone metastatic tissues. To detect the biological functions of LINC00482 in vitro, various assays were used including CCK-8, EdU, colony formation and transwell assays. The biological functions of LINC00482 were also identified in vivo by inoculating PCa cells into the left cardiac ventricle of mice, followed by evaluating the osteolytic lesions and osteolytic score. In addition, Starbase and Lncbase databases were applied for predicting the potential target miRNA of LINC00482, while TargetScan and Starbase databases were used for predicting the potential target of miRNA. The luciferase reporter assay was utilized to determine the interactions among these molecules and western blotting was employed to verified the targeted proteins. Results showed that high expression level of LINC00482 was observed in bone metastatic PCa tissues and associated with PCa progression. Silencing of LINC00482 inhibited cell proliferation, migration and invasion in PCa. Furthermore, LINC00482 was proved to act as a competing endogenous RNA by sponging miR-2467-3p to activate Wnt/ß-catenin signaling pathway, which may be a promising therapeutic target for PCa with bone metastasis.

Identification and Validation of the Prognostic Stemness Biomarkers in Bladder Cancer Bone Metastasis.

BACKGROUND: Bladder urothelial carcinoma (BLCA) is one of the most common urinary system malignancies with a high metastasis rate. Cancer stem cells (CSCs) play an important role in the occurrence and progression of BLCA, however, its roles in bone metastasis and the prognostic stemness biomarkers have not been identified in BLCA. METHOD: In order to identify the roles of CSC in the tumorigenesis, bone metastasis and prognosis of BLCA, the RNA sequencing data of patients with BLCA were retrieved from The Cancer Genome Atlas (TCGA) databases. The mRNA expression-based stemness index (mRNAsi) and the differential expressed genes (DEGs) were evaluated and identified. The associations between mRNAsi and the tumorigenesis, bone metastasis, clinical stage and overall survival (OS) were also established. The key prognostic stemness-related genes (PSRGs) were screened by Lasso regression, and based on them, the predict model was constructed. Its accuracy was tested by the area under the curve (AUC) of the receiver operator characteristic (ROC) curve and the risk score. Additionally, in order to explore the key regulatory network, the relationship among differentially expressing TFs, PSRGs, and absolute quantification of 50 hallmarks of cancer were also identified by Pearson correlation analysis. To verify the identified key TFs and PSRGs, their expression levels were identified by our clinical samples via immunohistochemistry (IHC). RESULTS: A total of 8,647 DEGs were identified between 411 primary BLCAs and 19 normal solid tissue samples. According to the clinical stage, mRNAsi and bone metastasis of BLCA, 2,383 stage-related DEGs, 3,680 stemness-related DEGs and 716 bone metastasis-associated DEGs were uncovered, respectively. Additionally, compared with normal tissue, mRNAsi was significantly upregulated in the primary BLCA and also associated with the prognosis (P = 0.016), bone metastasis (P < 0.001) and AJCC clinical stage (P < 0.001) of BLCA patients. A total of 20 PSRGs were further screened by Lasso regression, and based on them, we constructed the predict model with a relatively high accuracy (AUC: 0.699). Moreover, we found two key TFs (EPO, ARID3A), four key PRSGs (CACNA1E, LINC01356, CGA and SSX3) and five key hallmarks of cancer gene sets (DNA repair, myc targets, E2F targets, mTORC1 signaling and unfolded protein response) in the regulatory network. The tissue microarray of BLCA and BLCA bone metastasis also revealed high expression of the key TFs (EPO, ARID3A) and PRSGs (SSX3) in BLCA. CONCLUSION: Our study identifies mRNAsi as a reliable index in predicting the tumorigenesis, bone metastasis and prognosis of patients with BLCA and provides a well-applied model for predicting the OS for patients with BLCA based on 20 PSRGs. Besides, we also identified the regulatory network between key PSRGs and cancer gene sets in mediating the BLCA bone metastasis.

17ß-Estradiol/extrogen receptor ß alleviates apoptosis and enhances matrix biosynthesis of nucleus pulposus cells through regulating oxidative damage under a high glucose condition.

BACKGROUND: Hyperglycemia in the Diabetes mellitus (DM) patients is a potential etiology of disc degeneration. 17ß-estradiol (17ß-E2) supplementation plays an anti-diabetic role in DM patients. OBJECTIVE: This study was aimed to investigate the role and mechanism of 17ß-E2 in regulating nucleus pulposus (NP) cell apoptosis and NP matrix production under a high glucose condition. METHODS: Rat NP cells were cultured in medium with a high glucose concentration (0.2 M). 17ß-E2 was added into the culture medium to investigate its protective effects. The ERß inhibitor PHTPP and ERß activator ERB041 were used to investigate the effects of ERß. NP cell apoptosis was analyzed by flow cytometry and expression of apoptosis-related molecules. NP matrix production was evaluated by expression of matrix macromolecules. Additionally, intracellular reactive oxygen species (ROS) content was also detected. RESULTS: Compared with the control NP cells, 17ß-E2 decreased NP cell apoptosis ratio, down-regulated gene expression of Bax and caspase-3, up-regulated gene expression of Bcl-2, increased protein expression of cleaved PARP and cleaved caspase-3, and increased expression of matrix molecules (aggrecan and collagen II). Moreover, 17ß-E2 decreased ROS content. Further analysis showed that ERß inhibition partly reversed these effects of 17ß-E2 whereas ERß activation further promoted its effects. CONCLUSION: 17ß-E2/ERß interaction attenuates apoptosis and promotes matrix biosynthesis of NP cells through alleviating oxidative damage under a high glucose condition. This study provides new knowledge on strategies for retarding disc degeneration.

Heterogeneous expression and biological function of SOX18 in osteosaroma.

Osteosaroma (OS) is a primary bone malignancy and is associated with high morbidity. Sex determining region Y-box 18 (SOX18) is identified overexpressed in OS. However, the molecular mechanism underlying the biological function of SOX18 in OS is still unclear. The aim of the current study was to determine the SOX18 expression in patients with OS and its effect on tumor cell malignant phenotypes. Our results showed that SOX18 was overexpressed in OS patients from both E-MEXP-3628 database and independent samples from our hospital and in OS cell lines. SOX18 silencing significantly induced G0-G1 phase cell cycle arrest and apoptosis and inhibited U-2OS cell migration and invasion and cell growth both in vitro and in vivo. However, SOX18 overexpression remarkably promoted 143B cell proliferation, migration and invasion and inhibited cell cycle arrest and apoptosis. The protein expression levels of p53, p21, Bax, Bcl-2, and Caspase-3 were also regulated by SOX18. Moreover, SOX18 was found negative correlated with the expression of HERC1, HER2, HERC3, HERC4, HERC5, and HERC6 in OS patients and in OS cells, with the most significant correlation detected in HERC2 expression, which was following found interacted with SOX18 in OS cells. Taken together, our results suggest that SOX18 is overexpressed in OS and plays an important role in proliferation, apoptosis, migration and invasion of OS cells, and may provide a novel and promising thera-peutic strategy for OS.

Lumbar idiopathic intervertebral disc calcification associated with ossification of the ligamentum flavum in adult: a case report.

A 23 year-old female was diagnosed with lumbar idiopathic intervertebral disc calcification associated with ossification of OLF. The patient underwent posterior decompression and posterolateral fusion with pedicle screw fixation, which achieved excellent clinical improvement. Surgical intervention of a posterior approach for decompression and instrumented fusion is indicated in cases that the spinal cord compression with neurologic deficits was involved.

FS-3D-FISP for the diagnosis of ankle impingement syndrome and the evaluation of clinical outcomes of arthroscopic surgery.

This study aimed to assess the diagnostic value of three types of MRI sequences and to observe the clinical outcomes of arthroscopy surgery for ankle joint impingement syndrome. Ankle joint impingement syndrome was confirmed by FSE-T2WI, FSE-PDWI, and FS-3D-FISP MRI in 23 patients with arthroscopically proven ankle impingement. All 23 patients underwent arthroscopic surgery and the ankle joint function was evaluated before, 1 week after and 6 months after the operation. The patients were followed-up for 12-64 months (average 28 months). There was no significant difference in ankle function score between preoperatively and 1 week postoperatively, but 86.96 % patients got overall excellent or good scores 6 months after the surgery, significantly higher than before the surgery. The FS-3D-FISP MRI exhibited a good consistency with arthroscopic examination and had higher sensitivity and specificity for the diagnosis of ankle impingement than FSE-T2WI and FSE-PDWI. In summary, arthroscopy surgery for ankle impingement syndrome has several advantages such as good efficacy, minimal trauma, quick recovery, and much less complications. The preoperative FS-3D-FISP MRI allows accurate diagnosis and positioning of ankle impingement syndrome.

Effect of combination therapy with alginate dressing and mouse epidermal growth factor on epidermal stem cells in patients with refractory wounds.

BACKGROUND: The aim of this research was to determine the efficacy of combination therapy using an alginate dressing and mouse epidermal growth factor (mEGF) on proliferation and differentiation of epidermal stem cells (ESCs) in patients with refractory wounds. METHODS: Eighteen patients (12 males and 6 females, aged from 18 to 61 years (mean 36.4 years)) with various skin wounds, were treated by dressing changing for one month. The wounds were located in the foot (11), calf (3), thigh (2) and forearm (2). The patients were randomly divided into 3 groups: alginate dressing and mEGF (group A; n = 6), mEGF (group B; n = 6) and control (group C; n = 6). Wound closure indexes were measured at 7, 14, 21 and 28 days. Samples were harvested for pathologic examination, at 7 and 14 days following treatment. Cytokeratin 10 (CK10) and cytokeratin 15 (CK15) positive cells were evaluated using the super-sensitivity (SP) immunohistochemical staining technique. RESULTS: Wound healing was promoted in groups A and B. In group A, the wound closure index was increased significantly (P < 0.05), and in one case the maximum cure area reached 102 cm(2). Pathological examination identified a thicker epidermis, active angiogenesis and enhanced granulation in group A compared with groups B and C. Using the SP immunohistochemical staining technique, we showed that ESCs in group A were bigger in size and larger in number than in groups B and C. Overall, there was a significant difference in ESCs proliferation and differentiation between group A and group B (or C). CONCLUSIONS: Combination therapy using an alginate dressing and mEGF shows increased proliferation and differentiation of ESCs in patients with refractory wounds compared with those treated with mEGF alone.

[Application value of FS-3D-FISP sequence in the diagnosis of ankle cartilage injury].

OBJECTIVE: To compare several sequences of MRI and arthroscopy for detecting the ankle articular cartilage lesions and to evaluate the clinical outcome of special sequence of FS-3D-FISP. METHODS: Forty patients (41 ankles) with iterative ankle pain who were scheduled for arthroscopy underwent MR scanning, including FS-3D-FISP, FSE T2WI and FSE PDWI sequences. The results of each sequence were then compared with the arthroscopic findings. RESULTS: Using arthroscopic results as the standard of reference, the FS-3D-FISP images had the higher sensitivity (92.86%) than the other two sequences. The FS-3D-FISP sequence was well consistent with the result of arthroscopy. Kappa value (0.7590) was higher than the other two sequences (P < 0.01). CONCLUSION: As a favorable scanning sequence, FS-3D-FISP imaging can show the early-stage pathological changes of articular cartilage and it has an excellent correlation with the arthroscopic findings. A 3-D reconstruction is helpful to determine the location and the degree of lesion and obtain a more accurate classification to guide clinical decisions.

Functional protection of pentoxifylline against spinal cord ischemia/reperfusion injury in rabbits: necrosis and apoptosis effects.

BACKGROUND: Little is known about neuronal death mechanisms following spinal cord ischemia. The present study aimed to investigate the protective effect of pentoxifylline (PTX) against spinal cord ischemia/reperfusion (I/R) injury. METHODS: Rabbits sustained spinal cord ischemia following 45 minutes cross-clamping of the infrarenal aorta. Experimental groups were as follows: the first group of animals (sham, n = 8) underwent laparotomy alone and served as the sham group; the second group (I/R, n = 20) received carrier (3 ml saline solution) and served as the control group; the third group (PTX-A, n = 20) received PTX intravenously 10 minutes prior to ischemia; and the fourth group (PTX-B, n = 20) received PTX intravenously at the onset of reperfusion. Rabbits were evaluated for hind-limb motor function with the Tarlov scoring system at 48 hours. Serum was assayed with enzyme-linked immunosorbent assay for tumor necrosis factor alpha (TNF-alpha) and spinal cords were harvested for myeloperoxidase (MPO) activity, histopathological analysis, terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling staining, platelet/endothelial cell adhesion molecule-1 (PECAM-1) and caspase-3 immunohistochemistry, and the number of necrotic and apoptotic neuron were counted and data analyzed at 12, 24, 48 and 72 hours of reperfusion. Spinal cords were studied by electron microscopy. RESULTS: Improved Tarlov scores were seen in PTX-treated rabbits as compared with ischemic control rabbits at 48 hours. A significant reduction was found in TNF-alpha in serum, activity of MPO and immunoreactivity of the PECAM-1 and caspase-3 in PTX-treated rabbits. There were fewer apoptotic neurons than necrotic neurons (P < 0.05). A significant decrease in both necrotic and apoptotic neurons was observed in the PTX-treated groups (PTX-A and PTX-B) compared with the I/R group (P < 0.05). Both necrotic and apoptotic neurons were found with the electron microscope. CONCLUSIONS: PTX may induce protection against ischemia injury in the spinal cord, thereby preventing both necrosis and apoptosis. A major mode of cell death in spinal cord ischemia/reperfusion injury is necrosis while apoptosis is not dominant.