MiR-423-5p Regulates Cells Apoptosis and Extracellular Matrix Degradation via Nucleotide-Binding, Leucine-Rich Repeat Containing X1 (NLRX1) in Interleukin 1 beta (IL-1ß)-Induced Human Nucleus Pulposus Cells.

BACKGROUND Disc degeneration is characterized partly by the degradation in the extracellular matrix (ECM) and excess apoptosis of nucleus pulposus (NP) cells. NLRX1 (nucleotide-binding, leucine-rich repeat containing X1) is different from the other nucleotide-binding-domain and leucine-rich-repeat proteins and mainly located to the mitochondrial. It negatively regulates NF-κB (nuclear factor kappa B) and apoptosis inhibition. However, how NLRX1 is regulated and exerts effects in disc degeneration is unclear. Thus, the study aimed to analyze the effects of NLRX1 on NP cells. MATERIAL AND METHODS NLRX1 expression was detected in interleukin (IL)-1ß-induced NP cells by western blot and quantitative real-time polymerase chain reaction (qRT-PCR). Then, NLRX1 was overexpressed in IL-1ß-induced NP cells to detect apoptosis-related proteins and the extracellular matrix (ECM) by western blot, along with the detection of apoptosis levels using flow cytometry. StarBase predicted miR-423-5p target 3'UTR of NLRX1. Dual luciferase reporter assay showed that miR-423-5p could bind to the 3'UTR of NLRX1. Besides, miR-423-5p significantly affected NLRX1 levels detected by qRT-qPCR. RESULTS The miR-423-5p overexpression markedly, and negatively regulated the protective effects of NLRX1 on IL-1ß induced NP cells. Thus, our results suggested that miR-423-5p mediated the regulation of NLRX1 to affect apoptosis and ECM levels in IL-1ß induced NP cells. CONCLUSIONS miR-423-5p and NLRX1 could be potential therapeutic targets for patients with disc degeneration.

Outcomes of surgery for acromioclavicular joint dislocation using different angled hook plates: a prospective study.

PURPOSE: Hook plate fixation is widely used to treat acromioclavicular joint dislocation. However, there are many post-operative complications affecting the effect of treatment. The aim of this study is to evaluate the efficacy of the clavicular hook plate with different hook angles as a method of treatment in AC joint dislocation, and to guide the clinical application of hook plate. METHODS: We prospectively analysed 54 patients who were diagnosed with AC joint dislocation and treated with hook plate fixation by different hook angles. The patients were randomised into three groups: the -20° < AHP < 0° group, the 20° > AHP > 0° group and the 40° > AHP > 20° group. All patients were required to conform to regular follow-up post-operatively. Routine imaging to the shoulder was obtained to evaluate maintenance of the dislocation and the implant. Constant-Murley criteria were used to evaluate functional results. RESULTS: There were 19 patients in the -20° < AHP < 0° group, with one lost to follow-up, 22 patients in the 20° > AHP > 0° group, with two male patients lost to follow-up, and one female patient excluded because of no follow-up consent, and 19 patients in the 40° > AHP > 20° group, with one female and one male patient lost to follow-up. The Constant score was 61.8 ± 12.8, 74.7 ± 9.2 and 70.7 ± 9.4 before implant removal, and 78.8 ± 8.3, 87.1 ± 6.4 and 85.0 ± 6.1 after implant removal in the -20° < AHP < 0°, 20° > AHP > 0° and 40° > AHP > 20° groups, respectively. The functional results of the 20° > AHP > 0° and 40° > AHP > 20° groups were significantly better than the -20° < AHP < 0° group (P < 0.05), but the functional results of the 20° > AHP > 0° and 40° > AHP > 20° groups were not statistically significant. The CCD was 98.1 ± 4.8%, 107.5 ± 5.1% and 105.5 ± 4.1% before implant removal, and 98.8 ± 4.6%, 108.3 ± 4.8% and 107.2 ± 3.3% after implant removal in the three groups, respectively. The CCD of the 20° > AHP > 0° and 40° > AHP > 20° groups were statistically significantly different from the -20° < AHP < 0° group (P < 0.001). However, there was no statistical difference between the 20° > AHP > 0° group and the 40° > AHP > 20° group. Post-operative persistent pain occurred in 18.5% of all patients, post-operative stiffness occurred in 25.9% of all patients and 24.0% of patients had subacromial erosion. CONCLUSIONS: Hook plate treatment for AC joint dislocation can achieve the desired results, but the efficacy was significantly different depending on the different angles of the hook plate. AHP should be controlled within the range of 0-40° as much as possible when making clinical decisions.

Circular RNA circ_0081001 knockdown enhances methotrexate sensitivity in osteosarcoma cells by regulating miR-494-3p/TGM2 axis.

BACKGROUND: Circular RNAs (circRNAs) have been shown to participate in the chemoresistance and tumorigenesis of multiple cancers. The purpose of this research was to investigate the function of circ_0081001 in methotrexate (MTX) resistance of osteosarcoma (OS) and its potential molecular mechanism. METHODS: The expression of circ_0081001, cytochrome P450 family 51 subfamily A member 1 (CYP51A1), and miR-494-3p was detected by qRT-PCR. Cell viability, apoptosis, migration, and invasion were evaluated by Cell Counting Kit-8 (CCK-8) assay, flow cytometry, and transwell assay, respectively. Western blot (WB) assay was used to measure the protein levels of cleaved-caspase3 (cleaved-casp3), E-cadherin, N-cadherin, and transglutaminase-2 (TGM2). The interaction between miR-494-3p and circ_0081001 or TGM2 was predicted by bioinformatics analysis and verified using the dual-luciferase reporter assay. The mice xenograft model was established to investigate the roles of circ_0081001 in MTX resistance of OS in vivo. RESULTS: Circ_0081001 and TGM2 were upregulated, and miR-494-3p was downregulated in MTX-resistant OS tissues and cells. Moreover, circ_0081001 interference enhanced cell sensitivity to MTX through promoting apoptosis and inhibiting cell viability and metastasis in vitro. Furthermore, circ_0081001 was identified as a molecular sponge of miR-494-3p to upregulate TGM2 level. In addition, circ_0081001 knockdown inhibited MTX resistance via upregulating miR-494-3p and downregulating TGM2. Besides, circ_0081001 downregulation improved MTX sensitivity of OS in vivo. CONCLUSION: Knockdown of circ_0081001 enhanced MTX sensitivity of OS cells through downregulating TGM2 by sponging miR-494-3p, elucidating a novel regulatory mechanism for chemoresistance of OS and providing a potential circRNA-targeted therapy for OS.

CircSAMD4A contributes to cell doxorubicin resistance in osteosarcoma by regulating the miR-218-5p/KLF8 axis.

Circular RNA sterile alpha motif domain containing 4A (circSAMD4A) was found to be differentially expressed in osteosarcoma and contributed to the tumorigenesis of osteosarcoma. However, the role of circSAMD4A in doxorubicin (DXR) resistance of osteosarcoma is yet to be elucidated. Levels of circSAMD4A, microRNA (miR)-218-5p and Krüppel-like factor 8 (KLF8) were detected using quantitative reverse transcription-polymerase chain reaction. Western blot was applied to detect the protein levels of KLF8, cyclin D1 and p21. Cell viability, cell cycle, migration and invasion were analyzed using Cell Counting Kit-8 assay, flow cytometry and transwell assay, respectively. The interaction between miR-218-5p and circSAMD4A or KLF8 was verified using dual-luciferase reporter assay or RNA immunoprecipitation assay. In vivo experiments were performed using murine xenograft models. CircSAMD4A and KLF8 were elevated in osteosarcoma, and knockdown of circSAMD4A or KLF8 sensitized osteosarcoma cells to DXR by mediating resistant cell viability, migration and invasion inhibition, and cell cycle arrest in vitro. miR-218-5p was decreased in osteosarcoma, and miR-218-5p inhibition enhanced DXR resistance. Besides, miR-218-5p was found to bind to circSAMD4A or KLF8, and subsequent rescue experiments indicated that miR-218-5p inhibition reversed the inhibitory effects of circSAMD4A silencing on DXR resistance, and silencing miR-218-5p enhanced DXR resistance by targeting KLF8 in osteosarcoma cells. Moreover, circSAMD4A could indirectly regulate KLF8 via miR-218-5p. Additionally, circSAMD4A knockdown enhanced the cytotoxicity of DXR in osteosarcoma in vivo via regulating miR-218-5p and KLF8. In all, circSAMD4A enhanced cell DXR resistance in osteosarcoma by regulating the miR-218-5p/KLF8 axis, suggesting a novel therapeutic target for therapy-resistant osteosarcoma.

Popliteal pseudoaneurysm and arteriovenous fistula after arthroscopic anterior and posterior cruciate ligament reconstruction: A case report.

INTRODUCTION: Vascular injuries are a well recognised but very rare complication of surgery or trauma around the knee, especially associated with arthroscopic anterior and posterior cruciate ligament reconstruction. PRESENTATION OF CASE: This report describes a case of a Popliteal pseudoaneurysm and arteriovenous fistula after Arthroscopic anterior and Posterior Cruciate Ligament Reconstruction. An 57-year-old male who complained of pain and swelling around the left lower extremity after the ACL and PCL reconstruction using a semitendinosus graft from the two limbs caused by bicycle accident. He was diagnosed with the popliteal pseudoaneurysm and arteriovenous fistula by the Doppler ultrasonographic examination and CTA, and transferred to the interventional radiology suite where a successful vascular stent implantation was made. There was no postoperative complication. DISCUSSION: The most important finding of the present case were a popliteal pseudoaneurysm and arteriovenous fistula after the ACL and PCL reconstruction. To our knowledge, very few cases reporting an arteriovenous fistula with popliteal pseudoaneurysm following arthroscopy have been described. Vascular injury is a very rare complication of knee surgery, but surgeons should always consider this possibility in patients who have undergone knee arthroscopy. If the patient has symptoms of pain in the popliteal area and unexplainable swelling following arthroscopic ACL and PCL reconstruction, a popliteal vascular injury should be suspected. CONCLUSIONS: In order to minimize the risk of vascular complications in ACL and PCL reconstructive surgery, we advise to look for less traumatizing instruments and to limit the amount of riskful actions, precise attention should be paid in the establishment of the tibial tunnels.

[Treating Neer two- and three-part of proximal humeral fractures through anterolateral acromial approach and deltopectoral approach].

OBJECTIVE: To compare clinical results of treating Neer two- and three-part of proximal humeral fractures between anterolateral acromial approach and deltopectoral approach. METHODS: From January 2009 to December 2012, 49 patients with Neer two- and three-part of proximal humeral fractures were treated with locked plate fixation. In anterolateral acromial approach group, there were 22 patients including 9 males and 13 females with an average of (63.2±7.6) years old, while 27 patients in deltopectoral approach including 12 males and 15 females with an average of (62.9±7.0) years old. Operative time, blood loss during operation, fracture healing time and complications were observed and compared, postoperative Constant-Murley scoring and VAS scoring were applied for evaluate function of shoulder joint and pain at 3 months, 1 and 2 years respectively. RESULTS: All patients were followed up from 24 to 41 months with an average of 34.5 months. Operative time, blood loss, fracture healing time in anterolateral acromial approach group was (68.20±7.04) min, (151.30±20.57) ml, (10.88±4.90) weeks respectively, and better than that of in deltopectoral approach group which was (75.81±13.70) min, (242.10±37.25) ml and (13.60±2.45) weeks. Three months after operation, Constant-Murley scoring and VAS score in anterolateral acromial approach group was 88.32±5.45, 0.41±0.63 and better that of in deltopectoral approach group which was 63.53±8.31, 1.65±1.02. There was no significant differences between two groups in Constant-Murley scoring and VAS score at 1 and 2 years after operation. Each group has one case occurred loss of length humerus head height, and there was 1 case with subacromial impingement, 1 case with bolt loose and 2 cases with delayed union in deltopectoral approach. No axillary nerve injury, humeral head necrosis and breakage of internal fixation occurred both of two groups. CONCLUSION: Both of anterolateral acromial approach and deltopectoral approach are effective in treating Neer two- and three-part of proximal humeral fractures, and can obtain excellent outcomes. Moreover, anterolateral acromial approach has advantage of less trauma, less blood loss, shorter operative time, rapid recovery of shoulder joint function and fracture.

[Analysis and strategy on the early complications of lumbar disc herniation with Coflex system].

OBJECTIVE: To explore the short-term effectiveness and the cause of the early complications of lumbar disc herniation with Coflex system in order to provide evidence for the prevention. METHODS: From November 2007 to August 2008, 37 patients (20 males and 17 females) were treated with Coflex system. The age was from 33 to 70 years with an average of 52 years and the history was from 6 to 50 months with an average of 16.5 years. Complications were observed and the short-term effectiveness was evaluated by scores of JOA and ODI before and after operation. RESULTS: All patients were followed up from 1 to 2 years with an average 20 months. The JOA score increased from 9.09 +/- 1.10 preoperatively to 25.40 +/- 1.20 in the last follow-up. ODI decreased from 24.70 +/- 4.80 preoperatively to 4.80 +/- 1.00 in the last follow-up. The VAS score decreased from 7.86 +/- 0.80 preoperatively to 3.20 +/- 0.50 in the last follow-up. The symptoms remarkably improved. Complications occurred in 4 pa-tients (10.8%), among them, persistent low back pain was in 1 case and conservative treatment did not work; opposite lower limb pain was in 1 case at the 3rd week after operation and symptomatic treatment was effective; displacement of Coflex was in 1 case and Coflex breakage happened in 1 case at the 6th month after operation, but both did not have related clinical symptom. CONCLUSION: Coflex can obtain good clinical outcomes in treating lumbar disc herniation, but it has special complications. The indications and manipulations should be chosen properly.