LncRNA KCNQ1OT1 promoted BMP2 expression to regulate osteogenic differentiation by sponging miRNA-214.

BACKGROUND: Osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) is of much significance for bone formation, the imbalance of it would result in osteoporosis and other pathological bone defects. Increasing evidences showed that long non-coding RNAs (lncRNAs) and miRNAs played vital roles in the regulation of osteogenic differentiation. LncRNA KCNQ1OT1 was often regarded as an imprinted lncRNA and was related to tumor progression, while its function in osteogenic differentiation remained unclear. METHOD: qRT-PCR was performed to detect the expression of KCNQ1OT1, miR-214 and osteogenesis-related genes BMP2, Runx2, OPN, and OCN. Western blotting was carried out to detect osteogenesis-related markers. The osteoblastic phenotype was evidenced by alkaline phosphatase (ALP) activity and Alizarin Red S accumulation detection. Bioinformatics and luciferase assays were used to predict and validate the interaction between KCNQ1OT1 and miR-214 as well as BMP2 and miR-214. RESULTS: KCNQ1OT1 was significantly up-regulated during the process of osteogenic induction while miR-214 was contrarily down-regulated. Knockdown of KCNQ1OT1 inhibited osteogenic differentiation and down-regulated BMP2 and osteogenesis-related genes. It was also confirmed that KCNQ1OT1 directly interacted with miR-214. Meanwhile, miR-214 could bind to 3'UTR of BMP2 and therefore inhibited its expression. Furthermore, co-transfection of miR-214 inhibitor could rescue the down-regulation of BMP2 and osteogenesis-related genes and osteogenic differentiation suppression induced by KCNQ1OT1 knockdown. Moreover, miR-214 inhibitor significantly reversed the decreased protein levels of p-Smad1/5/8, Runx2 and Osterix induced by shKCNQ1OT1. CONCLUSIONS: KCNQ1OT1 positively regulated osteogenic differentiation of BMSCs by acting as a ceRNA to regulate BMP2 expression through sponging miR-214.

[Development of static three dimensional screw-plate system and the biomechanic features thereof, a comparative study].

OBJECTIVE: To develop a new internal fixation device and the matching tools for subtrochanteric fracture, investigate the biomechanical features thereof, and compare its effects with those of other instruments. METHODS: A new internal fixation device, static three dimensional screw-plate system: (STDSP) was developed based on the anatomical parameters and the biomechanical characteristics of proximal femur and combined with the biomechanical advantages of the model of three-dimensional plate and multiple-screws plate fixation. Sixteen paired cadaveric adult femurs were randomly divided into 2 equal groups: fracture healing group undergoing insertion of implant and then taking out thereof with a nail left, and fracture group with a trapezoid defect to mimic subtrochanteric fracture 4 of which were fixed with dynamic hip screw (DHS) and STDSP or Gamma nail and STDSP. All the specimens underwent A-P position four point bending test, axial compression test, torsion test, and destroyable test. RESULTS: (1) In the fracture group, A-P position four point bending test showed that the stretch displacement of STDSP group was (0.221 +/- 0.089) mm, significantly smaller than that of the Gamma nails group [(0.506 +/- 0.086) mm, t = 38.278 P = 0.000]; Axial compression test showed that the inferior displacement of the STDSP group was (2.804 +/- 0.585) mm, significantly larger than that of the Gamma nail group [(1.874 +/- 0.549) mm, t = -5.454, P = 0.012]; Torsion test showed that the torque values at 50 and 100 of the STDSP group were (3.78 +/- 0.84) J and (2.3 +/- 1.08) J, both significantly larger than those of the Gamma nail groups [(2.3 +/- 1.08) J and (4.35 +/- 1.81) J, t = -4.729 P = 0.018, and t = -3.227 P = 0.048] and those of the DHS group [(7.3 +/- 1.273) J and (4.325 +/- 0.88) J respectively, t = -3.242 P = 0.048]; and torsion destroyable test showed that the torque values at 200 of the STDSP group was (21.88 +/- 2.51) J, significantly larger than those of the Gamma nail and DHS groups, [(14.36 +/- 5.01) J and (15.475 +/- 1.924) J and (12.075 +/- 1.179) J, t = -3.323 P = 0.048, t = - 3.732 P = 0.034]. (2) In the fracture healing specimens, A-P position four point bending test showed that the radial degree displacement of the STDSP group was (1. 050 +/- 0.316) mm, significantly smaller than that of the DHS group [(1.261 +/- 0.364) mm, t = 3.4 P = 0.042]. CONCLUSIONS: More capable of resisting bending than DHS and less capable of resisting axial compression, STDSP is an excellent internal fixator with good biomechanical feature in treating subtrochanteric femoral fracture.

[Protein expression of Skp2 in osteosarcoma and its relation with prognosis].

OBJECTIVE: To investigate the expression of Skp2 and its relation with P27 expression, clinic pathologic features, and prognostic indicator in osteosarcoma. METHODS: We collected osteosarcoma specimen from 52 patients (29 males and 23 females), who were all treated by radical resection of tumor. The expression of Skp2 and P27 was determined by SP immunohistochemistry. Forty-four patients were followed up for 4 to approximately 84(mean = 31.2)months, while the other 8 patients were lost. Twenty of them survived over 5 years and 24 died. RESULTS: In osteosarcoma, Skp2 highly expressed (mean value was 1.74). Expression intensity of Skp2 at the stage III was obviously higher than that of the stage II(IIa and IIb) (P < 0.05). Skp2 expression was correlated with the relapse, metastasis, and 5-year survival in osteosarcoma (P < 0.05), but not with different pathologic types, sex, or age(P > 0.05). The expressions of skp2 and P27 were negative correlation in osteosarcoma (r = -0.907, P < 0.05). CONCLUSION: Skp2 plays an important role in the occurrence and development of osteocarcoma by causing the degradation of P27, and can be an important prognostic indicator in osteosarcoma.

[Internal fixation with T type titanium plate for volar Barton's fracture].

OBJECTIVE: To evaluate the operative method and the clinical efficacy of volar Barton's fracture treated by internal fixation with T type titanium plate. METHODS: We performed the operation of the open reduction and internal fixation with T type titanium plate in 18 patients with volar Barton's fracture. RESULTS: The 18 patients were followed up after the operation at the average time of 13 months (6 to 31 months). X radiographs showed that all patients achieved solid bone union. Mehara's evaluation for volar Barton's fracture revealed excellent outcome in 12 patients, good in 5 patients, and fair in 1 patients. The excellent and good rate of clinical results was 94.4%. X-radiographs showed that the volar tilting angle was 8-18 (mean=11.0) degree, and the ulnar inclining angle was 16-26 (mean=22.4) degree. There was no shortening of the radius. CONCLUSION: The operative treatment with titanium T type plate fixation for volar Barton's fracture is in favor of fracture anatomic reduction, solid internal fixation, and catagmatic concrescence. It is beneficial for functional recovery of the wrist at the utmost. The outcome in the near future is satisfactory. It is an ideal method for volar Barton's fracture.

[Design and manufacture of the static 3 dimensional screw-plate system and its matching tools].

OBJECTIVE: To design and manufacture a new type instrument and its matching tools for the proximal femur. METHODS: A new type instrument of subtrochanterical fracture-static 3-dimensional screw-plate system(STDSP)was designed and manufactured. The different types of accessory instruments which adapted to STDSP,the aiming guide, and the accessory tools were also designed and manufactured, and they were tested on bones. RESULTS: STDSP included 3 special lag screws and 1 anatomic plate. The lag screws distributed in femoral neck like "or"shape. There was 135 degree between the lag screws and femoral shaft. The shape of the plate was similar to the anatomic structure of the proximal thighbone. The posterior screw was stably connected to the plate. The plate was affixed crosswise to the lateral and anterior side by bicortical screws. The aiming guides and all kinds of accessory tools could help to install STDSP exactly and conveniently,which was verified on cadavers and X-ray photographs. CONCLUSION: STDSP has the function of dynamic compressing in 3-dimensional space, and can match with the structure of the proximal femur preferably. STDSP may be a suitable instrument to the subtrochanteric fractures. The accessory apparatus of STDSP has simple configuration,and can let the STDSP be installed accurately, conveniently and rapidly.

[Integrity of lumbar facet joint and curative effect after a lumbar intervertebral disc operation].

OBJECTIVE: To analyze the relation between the clinical outcome and the integrity of the facets after a lumbar operation,and to provide a reference for choosing operative method and clinical prognosis. METHODS: Forty-three patients with complete data underwent uni-segment discectomy were enrolled. There were 3 surgical interventions: open-window discectomy, full or semi-laminectomy. Groups were divided based on the integrity of the facets after the operation, and the clinical symptoms and signs were evaluated using the Japanese Orthopaedic Association Back (JOA) scores at 24-month follow-up. RESULTS: Preoperative JOA scores were not significantly different among the groups (P>0.05). Compared with the facet intact group at 24-month follow-up,JOA scores were descended statistically in total uni-facetectomy group and total uni-facetectomy plus partial opposite facetectomy group (P<0.01). CONCLUSION: Keeping facets integrated plays an important role in achieving good clinical results,and the damage of facet should be avoided in the lumbar operation.

[Clinical analysis of the treatment of reserving limb for patients with malignant bone tumor].

OBJECTIVE: To investigate the curative effect of the treatment of reserving limb for patients with malignant bone tumor. METHODS: Forty patients with bone neoplasms underwent the operation of reserving limbs. The neoadjuvant chemotherapy was given to all the patients before the operation. After the ablation of the tumor we used the artificial prosthetic replacement,tumor tissue deactivation and reimplant, variant bone and joint transplantation to reserve the limb. RESULTS: All the patients were followed up. The average follow-up time was 34.8 months. Eight patients got complications. And the fineness rate was 72.5% according to functional evaluation standard of Enneking after surgical treatment of bone tumor. CONCLUSION: The operation of reserving limb was an ideal method to cure the malignant bone tumor. The proper operative treatment with neoadjuvant chemo-therapy is helpful to extend the life of the patients and increase the quality of life.

[Clavicular hook plate in the treatment of dislocation of acromioclavicular joint and fracture of distal clavicle].

OBJECTIVE: To evaluate the curative effects of AO clavicular hook plate in the treatment of dislocation of acromioclavicular joint and fracture of distal clavicle. METHODS: We retrospectively analyzed the clinical data of 39 patients with acromioclavicular joint dislocation or distal clavicle fractures treated with clavicular hook plate: 16 suffered from dislocation of acromioclaviculr joint, 18 suffered from far end collarbone fractures, and 5 patients had both symptoms. RESULTS: The 39 patients were followed up for 12 months averagely. All patients got good reduction and fixation. The acromioclavicular joint subluxation, breakage, and loosening of the clavicular hook plate were not observed. The acromioclavicular joint recovered quickly. CONCLUSION: AO clavicular hook plate is a good choice for acromioclavicular joint dislocation and distal clavicular fracture.

Differential effects of MTSS1 on invasion and proliferation in subtypes of non-small cell lung cancer cells.

Non-small cell lung cancer (NSCLC) accounts for >80% of all cases of lung cancer and can be divided into lung adenocarcinoma (LAC), large-cell carcinoma (LCC), and squamous cell carcinoma (SCC). Accumulating evidence suggests that MTSS1, which is a newly discovered protein associated with tumor progression and metastasis, may have differential roles in cancer malignancy. As it has been demonstrated that MTSS1 is overexpressed in NSCLC and may be an independent prognostic factor in patients with SCC, the present study explored the differential roles of MTSS1 in the invasion and proliferation of different subtypes of NSCLC. Stable overexpression and knockdown of MTSS1 was performed in human NSCLC H920 (LAC), H1581 (LCC) and SW900 cell lines (SCC), and western blot, cell invasion, proliferation and FAK activity analyses were used to investigate the effects. Overexpression of MTSS1 enhanced the invasion and proliferation abilities of H920 and H1581 cells, and these effects were abolished by treatment with selective FAK inhibitor 14, which did not affect the expression of MTSS1. Notably, overexpression of MTSS1 inhibited invasion and proliferation in SW900 cells, and this effect was enhanced by the selective FAK inhibitor. Knockdown of MTSS1 decreased the invasion and proliferation abilities of H920 and H1581 cells, whereas knockdown increased invasion and proliferation in SW900 cells. Furthermore, while overexpression of MTSS1 induced FAK phosphorylation and activity in H920 and H1581 cells, MTSS1 overexpression inhibited FAK phosphorylation/activity in SW900 cells. Knockdown of MTSS1 decreased FAK phosphorylation/activity in H920 and H1581 cells, whereas knockdown increased these processes in SW900 cells. To the best of our knowledge, the present study was the first to demonstrate that MTSS1 has differential roles in various subtypes of NSCLC, acting via a FAK-dependent mechanism. The results indicated that MTSS1 may enhance invasion and proliferation in LAC and LCC cells, whereas MTS11 inhibits these processes in SCC cells. These findings provide novel insight into the functional role of MTSS1 in cancer and may help elucidate therapeutic strategies for the treatment of various types of cancer.

Decreased expression of SLC 39A14 is associated with tumor aggressiveness and biochemical recurrence of human prostate cancer.

OBJECTIVE: Solute carrier family 39, member 14 (SLC39A14), has been identified as a potential biomarker for various cancers. However, its roles in prostate cancer (PCa) are still unclear. The aim of this study was to investigate the clinical significance of SLC39A14 in patients with PCa and its functions in malignant phenotypes of PCa cells. PATIENTS AND METHODS: Subcellular localization and expression pattern of SLC39A14 protein were examined by immunohistochemistry. Then, the associations of SLC39A14 expression with various clinicopathological features and clinical outcome of patients with PCa were statistically evaluated. Subsequently, the effects of SLC39A14 overexpression and knockdown on PCa cell proliferation and motility were, respectively, examined by Cell Counting Kit-8, transwell, and wound-healing assays. RESULTS: The immunoreactive scores of SLC39A14 protein in human PCa tissues were significantly lower than those in normal prostate tissues. Based on the Taylor dataset, SLC39A14 downregulation occurred more frequently in patients with PCa with a higher Gleason score (P<0.001), advanced clinical stage (P=0.008), presence of metastasis (P=0.009), and prostate-specific antigen failure (P=0.006). More interestingly, the survival analysis identified SLC39A14 as an independent factor for predicting the biochemical recurrence-free survival of patients with PCa (P=0.017). Functionally, the enforced expression of SLC39A14 could suppress cell proliferation, invasion, and migration of PCa cell lines in vitro, which could be reversed by the knockdown of SLC39A14. CONCLUSION: Decreased expression of SLC39A14 may lead to malignant phenotypes of PCa cells and aggressive tumor progression in patients with PCa. Importantly, SLC39A14 may function as a tumor suppressor and a biomarker for screening patients with biochemical recurrence following radical prostatectomy.

Biliary Cast Syndrome: Hepatic Artery Resistance Index, Pathological Changes, Morphology and Endoscopic Therapy.

BACKGROUND: Biliary cast syndrome (BCS) was a postoperative complication of orthotopic liver transplantation (OLT), and the reason for BSC was considered to relate with ischemic type biliary lesions. This study aimed to evaluate the relationship between BCS following OLT and the hepatic artery resistance index (HARI), and to observe pathological changes and morphology of biliary casts. METHODS: Totally, 18 patients were diagnosed with BCS by cholangiography following OLT using choledochoscope or endoscopic retrograde cholangiopancreatography. In addition, 36 patients who did not present with BCS in the corresponding period had detectable postoperative HARI on weeks 1, 2, 3 shown by color Doppler flow imaging. The compositions of biliary casts were analyzed by pathological examination and scanning electron microscopy. RESULTS: HARI values of the BCS group were significantly decreased as compared with the non-BCS group on postoperative weeks 2 and 3 (P < 0.05). Odds ratio (OR) analysis of HARI 1, HARI 2, HARI 3 following the operation was >1 (OR = 1.300; 1.223; and 1.889, respectively). The OR of HARI 3 was statistically significant (OR = 1.889; 95% confidence interval = 1.166-7.490; P = 0.024). The compositions of biliary casts were different when bile duct stones were present. Furthermore, vascular epithelial cells were found by pathological examination in biliary casts. CONCLUSIONS: HARI may possibly serve as an independent risk factor and early predictive factor of BCS. Components and formation of biliary casts and bile duct stones are different.

MicroRNA-145 inhibits lung cancer cell metastasis.

Previous studies have identified a variety of microRNAs (miRNAs) that have important roles in cancer progression, particularly in tumor invasion and metastasis. Downregulation of miR­145 was reported to occur in various types of human cancer; however, the role of miR­145 in lung cancer metastasis and its potential mechanisms of action remain to be elucidated. The present study aimed to investigate the effects of miR­145 on metastasis and epithelial­mesenchymal transition (EMT) in A549 human lung adenocarcinoma cells. In addition, the underlying mechanisms by which miR­145 regulates EMT were examined. The miR­145 mimic was transfected into A549 cells; cell invasion and adhesion assays were then performed in order to investigate cell metastasis, and western blot analysis was used to examine the expression of EMT markers. In order to further examine the underlying mechanisms by which miR­145 regulates EMT, a luciferase reporter assay was performed to determine whether miR­145 targeted Oct4. In addition, the expression of Wnt3a and ß­catenin in A549 cells was measured following transfection with small hairpin RNA­Oct4. To the best of our knowledge, the results of the present study demonstrated for the first time, that miR­145 inhibited lung cancer cell metastasis and EMT via targeting the Oct4 mediated Wnt/ß­catenin signaling pathway.

An atypical monomelic presentation of Mazabraud syndrome.

Mazabraud syndrome is a rare condition characterized by a combination of fibrous dysplasia and intramuscular myxomas. In Mazabraud syndrome, the distribution of fibrous dysplasia is mostly polyomelic and frequently located in the femur, with myxomas adjacent to the fibrous dysplasia lesion of bone (mostly in the quadriceps muscle). However, when presented as atypical clinical features, patients of Mazabraud syndrome is either misdiagnosed or difficult to diagnose. We report an atypical monomelic case of Mazabraud syndrome in the right upper arm and discuss the difficulties in making an accurate diagnosis.

Potential use of porous titanium-niobium alloy in orthopedic implants: preparation and experimental study of its biocompatibility in vitro.

BACKGROUND: The improvement of bone ingrowth into prosthesis and enhancement of the combination of the range between the bone and prosthesis are important for long-term stability of artificial joints. They are the focus of research on uncemented artificial joints. Porous materials can be of potential use to solve these problems. OBJECTIVES/PURPOSES: This research aims to observe the characteristics of the new porous Ti-25Nb alloy and its biocompatibility in vitro, and to provide basic experimental evidence for the development of new porous prostheses or bone implants for bone tissue regeneration. METHODS: The Ti-25Nb alloys with different porosities were fabricated using powder metallurgy. The alloys were then evaluated based on several characteristics, such as mechanical properties, purity, pore size, and porosity. To evaluate biocompatibility, the specimens were subjected to methylthiazol tetrazolium (MTT) colorimetric assay, cell adhesion and proliferation assay using acridine staining, scanning electron microscopy, and detection of inflammation factor interleukin-6 (IL-6). RESULTS: The porous Ti-25Nb alloy with interconnected pores had a pore size of 200 µm to 500 µm, which was favorable for bone ingrowth. The compressive strength of the alloy was similar to that of cortical bone, while with the elastic modulus closer to cancellous bone. MTT assay showed that the alloy had no adverse reaction to rabbit bone marrow mesenchymal stem cells, with a toxicity level of 0 to 1. Cell adhesion and proliferation experiments showed excellent cell growth on the surface and inside the pores of the alloy. According to the IL-6 levels, the alloy did not cause any obvious inflammatory response. CONCLUSION: All porous Ti-25Nb alloys showed good biocompatibility regardless of the percentage of porosity. The basic requirement of clinical orthopedic implants was satisfied, which made the alloy a good prospect for biomedical application. The alloy with 70% porosity had the optimum mechanical properties, as well as suitable pore size and porosity, which allowed more bone ingrowth.

Could nonunion tissue be transformed capable of bone formation by negative pressure: a new alternative to treat bone nonunion?

Despite substantial advances in orthopaedic surgery, bone nonunion is still a matter of debate for orthopaedic surgeon to provide optimal options on treatment of nonunion. Negative pressure therapy has already been successfully used in dealing with complex kinds of soft tissue healing. Some studies show that negative pressure can induce mesenchymal stem cells to differentiate into osteoblasts and others suggest that there are some mesenchymal-like cells existing in the nonunion tissue which can be re-activated and transformed into osteoblasts under some circumstance. We hypothesized that under negative pressure the mesenchymal-like cells can be transformed into osteoblasts in nonunion site. Under negative pressure, both "seeds" and "soils" can be made for new bone formation. If our hypothesis is proved to be corrected, it could be an innovative method to treat the bone nonunion.

Reduced expression of Raf kinase inhibitor protein correlates with poor prognosis in pancreatic cancer.

AIM: Raf kinase inhibitory protein (RKIP) is an inhibitor of the Raf/MEK/MAP kinase signaling cascade and a suppressor of cancer metastasis. But its function in pancreatic cancer was not yet clarified completely. The aim of this study was to investigate the involvement of RKIP in pancreatic cancer. METHODS: RKIP expression was investigated retrospectively by immunohistochemistry in paraffin-embedded primary tumor tissue samples from a series (n = 99) of consecutive patients with pancreatic cancer. Survival was calculated using Kaplan-Meier curves. Parameters found to be of prognostic significance in univariate analysis were verified in a multivariate Cox regression model. RESULTS: RKIP expression was high in normal pancreatic epithelium and retained to varying degrees in pancreatic cancer tissues. However, in tumor tissues with lymph node metastasis (P = 0.008) and high UICC stage (P = 0.006), RKIP expression was highly significantly reduced or lost. Furthermore, the reduced expression of RKIP significantly correlated with both poor overall and disease-free survival (P = 0.008 and 0.01, respectively). Multivariate analyses revealed RKIP to be an independent prognosticator. CONCLUSION: These findings suggest that RKIP could be a promising marker for predicting a better prognosis in pancreatic cancer.