Lnc-SELPLG-2:1 enhanced osteosarcoma oncogenesis via hsa-miR-10a-5p and the BTRC cascade.

BACKGROUND: To investigate the potential role of Long Non-coding RNAs (lncRNAs) in the progression of osteosarcoma. METHODS: The candidate lncRNAs were screened with RNA-seq and confirmed with quantitative real-time PCR. Using MTS, transwell assay, and flow cytometric analysis, the effects of overexpressed lnc-SELPLG-2:1 on cell functions were determined. Immunohistochemical staining, fluorescence in situ hybridization, and luciferase reporter assay were used to evaluate the potential mechanism of lnc-SELPLG-2:1 in vivo and in vitro using a tumor model. Moreover, the effects of overexpression of hsa-miR-10a-5p on the functions of SaOS2 cells were determined using functional cell analysis. A response test was used to confirm the mechanism by which lnc-SELPLG-2:1 sponge hsa-miR-10a-5p promotes the expression of BTRC to regulate osteosarcoma. RESULTS: Lnc-SELPLG-2:1 was highly expressed in osteosarcoma compared to normal cells and bone and marrow samples. Inhibition of lnc-SELPLG-2:1 accelerated cell apoptosis and suppressed cell proliferation, migration, and invasion, whereas lnc-SELPLG-2:1 overexpression had the opposite effect. Moreover, inhibiting lnc-SELPLG-2:1 in an in vivo model decreased tumor size and suppressed the expression of cell migration-related proteins. The prediction, dual luciferase assay, and response test results indicated that hsa-miR-10-5p and BTRC were involved in the lnc-SELPLG-2:1 cascade. Unlike lnc-SELPLG-2:1, hsa-hsa-miR-10a-5p had opposite expression and function. Competitive binding of lnc-SELPLG-2:1 to hsa-hsa-miR-10a-5p prevented BTRC from miRNA-mediated degradation, thereby activating the expression of VIM, MMP9, and MMP2, promoting osteosarcoma cell proliferation, migration, and invasion, and inhibiting apoptosis. CONCLUSION: Lnc-SELPLG-2:1 is an oncogenesis activator in osteosarcoma, and its functions are performed via hsa-miR-10a-5p /BTRC cascade.

Reduction of Blood Loss by Intra-articular Injection of Tranexamic Acid Combined with Knee and Hip Flexion at 45° During Primary Total Knee Arthroplasty: A Randomized Controlled Trial.

OBJECTIVE: To explore the hemostatic effect of intra-articular administration of tranexamic acid (TXA) combined with knee flexion in total knee arthroplasty (TKA). METHODS: This randomized controlled trial was conducted at the Third Affiliated Hospital of Southern Medical University (Guangzhou, China) from January 2017 to February 2018. The patients were randomized 1:1 to the TXA group (TXA 500 mg into the joint after closure, knee, and hip flexed at 45° for 4 h) or the control group (physiological saline, with limb fully extended). The primary endpoint was postoperative hemoglobin reduction. The postoperative levels of hemoglobin were measured at four time points: 6 h after operation, and on the first, second, and third postoperative days. Calculated blood loss (CBL) at 3 days, transfusion rate, range of motion (ROM), VAS pain score, and knee circumference increment were the secondary endpoints. Ninety-four (47/group) patients were analyzed. RESULTS: Postoperatively, there were statistically significant differences between the TXA and control groups in CBL (791 ± 212 mL vs 1175 ± 273 mL, P < 0.05). Hemoglobin reduction was significantly lower in the TXA group (2.0 ± 0.9 g/dL vs 4.5 ± 0.7 g/dL, P < 0.05). Based on the transfusion criteria, 3 out of 47 (6.4%) patients in the TXA group and 13 out of 47 (27.6%) patients in the control group received blood transfusions (P = 0.006). ROM (90.8° ± 6.2° vs 87.6° ± 6.4°, P = 0.004), VAS pain score (4.1 ± 1.1 vs 4.8 ± 1.3, P = 0.004), and KCI (2.4 ± 0.9 cm vs 3.2 ± 1.0 cm, P = 0.01) were better in the TXA group compared with thecontrols. There was no deep venous thrombosis (DVT), wound infection or other adverse events in either group. In the control group, 2 patients had a fever after blood transfusion. CONCLUSION: Intra-articular injection of TXA combined with knee and hip flexion at 45° can effectively attenuate CBL and hemoglobin reduction during primary TKA, without an additional adverse event.

Hsa_circ_0008934 promotes the proliferation and migration of osteosarcoma cells by targeting miR-145-5p to enhance E2F3 expression.

OBJECTIVE: To investigate the role of hsa_circ_0008934 in osteosarcoma and the molecular mechanism involved in the regulation of the occurrence and development of osteosarcoma METHODS: Differentially expressed circRNAs in the osteosarcoma cell lines SaOS2 and MG63 and in the normal human osteoblast cell line hFOB1.19 were identified via next-generation RNA sequencing. The expression and circular morphology of hsa_circ_0008934 were analyzed via quantitative real-time polymerase chain reaction (qRT-PCR) and RT-PCR analysis, respectively. Proliferation, apoptosis, cell cycle progression, migration, and invasion of SaOS2 and MG63 cells with hsa_circ_0008934 silencing or overexpression were assessed using the MTS method, colony formation assay, flow cytometry, and the transwell system, respectively. The subcellular distribution of hsa_circ_0008934 was revealed via fluorescence in situ hybridization. The binding of hsa_circ_0008934 with microRNAs was confirmed using the dual-luciferase reporter assay. The oncogenic roles of hsa_circ_0008934 in osteosarcoma were determined using an in vivo tumorigenesis assay with nude mice. qRT-PCR, western blotting, TUNEL assay, and immunohistochemistry (IHC) were used to detect the tumorigenicity of hsa_circ_0008934 in osteosarcoma cells. RESULTS: Many circRNAs were differentially expressed in SaOS2 and MG63 cells than in hFOB1.19 cells. Hsa_circ_0008934 expression was significantly elevated in SaOS2 and MG63 cells. Hsa_circ_0008934 silencing significantly reduced proliferation, enhanced apoptosis, blocked cell cycle progression, and impaired migration and invasion capacities of SaOS2 cells. Opposite cellular alterations were achieved by overexpressing hsa_circ_0008934 in MG63 cells. Hsa_circ_0008934 was mainly distributed in the cytosol and positively regulated E2F3 expression in osteosarcoma cells. In addition, it directly bound with miR-145-5p to repress E2F3 expression and enhanced the tumorigenesis of MG63 cells in nude mice. qRT-PCR revealed that the intracellular injection of hsa_circ_0008934 lentivirus resulted in hsa_circ_0008934 overexpression and miR-145-5p downregulation. Western blotting confirmed that E2F3 was upregulated. Moreover, the TUNEL assay showed that hsa_circ_0008934 overexpression inhibited the apoptosis of tumor cells. IHC detection revealed that the hsa_circ_0008934 overexpression could promote the expression of Ki67 and PCNA. CONCLUSION: Elevated hsa_circ_0008934 expression promotes the proliferation and migration of osteosarcoma cells by sponging miR-145-5p to enhance E2F3 expression.

EDNRB Reverses Methylprednisolone-Mediated Decrease in Neural Progenitor Cell Viability via Regulating PI3K/Akt Pathway and lncRNA Expression.

OBJECTIVE: To investigate the functions and mechanisms of methylprednisolone (MP) through endothelin receptor B (EDNRB) on the cell proliferation of neural progenitor cells (NPCs) to regulate spinal cord injury. METHODS: Primary NPCs were isolated from fetal mice and subjected to treatments with MP and IRL-1620 (EDNRB agonist). The cell viability was determined using the MTS assay. Total RNA was extracted from the cells, and RNA-seq was performed to screen for lncRNAs. The targets of the candidate lncRNAs were predicted by GO and KEGG analyses, and the expressions of lncRNAs were validated via qPCR. Furthermore, protein levels of the PI3K-AKT pathway were determined via Western blotting, and the expression of lncRNAs was detected after inhibiting the pathway with AKT inhibitor. RESULTS: MTS assays revealed that MP decreased the cell viability of NPCs, whereas the EDNRB agonist reversed this effect of MP. NPCs were used for RNA-seq in the following three groups: normal control (NC), MP, and MP combined with EDNRB agonist (MP + EDNRB). Our results suggested that the NONRATT030699.2, NONRATT004088.2, and NONRATT005601.2 lncRNAs might be involved in the signaling pathway that is correlated to MP and the EDNRB agonist. GO and KEGG pathway analyses revealed that this was the PI3K/AKT pathway. The relevant genes involved in the pathway were validated by Western blotting. The EDNRB agonist promoted cell proliferation mainly via the activation of the PI3K/AKT pathway; however, it suppressed the expression of p-ERK, thereby increasing the expression of cyclin D1 and attenuating the effect of MP in suppressing cell proliferation. Meanwhile, after the AKT signal pathway was inhibited, these lncRNA expressions were consistent with those in the MP + EDNRB group. CONCLUSION: MP inhibits NPC proliferation, whereas EDNRB activation reverses the effect of MP via lncRNA.

[Less invasive fixation for the treatment of comminuted fracture of distal tibia].

OBJECTIVE: To study the result of less invasive fixation in treating comminuted fracture of distal tibia. METHODS: From 2002 to 2008, 48 patients with comminuted fracture of distal tibia were treated with surgery. The closed reduction and less invasive fixation were done to stable broken fibula, and the reduction by Kirschner wire to pick was used for relatively larger debris of tibia. The Kirschner wire or screw were used to fix fracture after the restore of the ankle cavity position and the alignment of the tibia. Partial weight loading and functional exercise of ankle joint were done at 6th week after operation. RESULTS: Forty-eight patients were followed up for 1-24 months with an average of 12 months. All the fractures were united. According to Johner-Wruhs standard to value the result by factors of pain, deformity, motion range of joint,with or without injury of nerve and blood vessel. Thirty-eight cases obtained excellent result, 8 good, fair 2. The rate of excellent and good were 95.8%. CONCLUSION: Less invasive fixation has ascendancy such as easy operation, less injury of soft tissue, reliable fixation, which can maximally protect periosteum. It is a choice for treating comminuted fracture of distal tibia.

[Treatment of floating knee injuries with interlocked intramedullary nailing through a single incision].

OBJECTIVE: To investigate the technique and clinical effect of interlocked intramedullary nailing through a single incision on the treatment of floating knee injuries. METHODS: Thirty-eight patients of floating knee injuries included 25 male and 13 female with an average age of 35.6 years ranging from 17 to 52 years. All patients were treated by interlocked intramedullary nailing through a single incision. The clinical effect was evaluated postoperatively. RESULTS: Thirty-eight cases were followed-up for 8.6 to 15.4 months with an average of 12.0 months. Thirty-seven cases gained bone healing and the time of fracture healing was from 2.7 to 6.9 months, with an average of 4.8 months. The nonunion of the distal femoral fracture was observed in 1 case with retrograde nailing, the revision operation was performed by plate and bone graft, and bony union was obtained. The results were excellent in 22 cases, good in 13 cases, fair in 2 cases, and poor in 1 case. CONCLUSION: The treatment of floating knee injuries with interlocked intramedullary nailing through a single incision brings less damage, low infection rate, good fracture healing. It is an effective method for treating floating knee injuries.

[Different approaches of vertebroplasty for management of severe osteoporotic vertebral compression fractures].

OBJECTIVE: To compare the clinical efficacy of 3 approaches of vertebroplasty in the treatment of severe osteoporotic vertebral compression fractures. METHODS: Twenty-five patients with severe osteoporotic vertebral compression fractures were observed, whose average age was 72 years with average disease history of 12 days and average compression of the affected vertebral bodies of 73%. The patients were divided into 3 groups for 3 different fracture types according to Rao's classification of osteoporotic vertebral structure and deformity, namely wedge type (group A, n= 12), biconcave type (group B, n= 7) and crush type (group C, n=6). Unipedicular approach was adopted in group A, far lateral bipedicular approach in group B, and posterior wall vertebroplasty and pedicle screw fixation in group C. The average follow-up time was 1 year. Visual analog scale (VAS), analgesic use and the mobility were measured in the patients preoperatively and 3 days (7 days in group C) and 6 months after the operation, respectively, and the success rates and complications were observed. RESULTS: All the surgical procedures were successful. The average operation time was 35 min in group A, 50 min in group B, and 2 h in group C. The average volume of cement injected into each vertebral body was 2.0 ml. The average blood loss was 30 ml in groups A and B, and 600 ml in group C. The procedure increased mobility and decreased analgesic use. VAS was decreased by a mean of 4.8 in Group A, 6.2 in group B 3 days after the operation and 5.4 in group C 7 days postoperatively, and remained stable till 6 months after the operation (P<0.01). Cement extravasation occurred in 9 cases, cement in the vertebral canal in 2 cases with transient neurological symptoms, and cement extravasation in the intervertebral space and the anterior space of the vertebral bodies took place in 5 and 2 cases, respectively. No neurological complications were observed. CONCLUSION: Different types of severe osteoporotic vertebral compression fractures require management with different approaches of vertebroplasty for adequate filling of the remaining vertebral body, which provides significant pain relief with wider indications.