Revision surgery for periprosthetic fracture of distal femur after endoprosthetic replacement of knee joint following resection of osteosarcoma.

Purpose: Periprosthetic fracture (PPF) is one of the severe complications in patients with osteosarcoma and carries the risk of limb loss. This study describes the characteristics, treatment strategies, and outcomes of this complication. Methods: Patients were consecutively included who were treated at our institution between 2016 and 2020 with a PPF of distal femur. The treatment strategies included two types: 1) open reduction and internal fixation with plates and screws and 2) replacement with long-stem endoprosthesis and reinforcement with wire rope if necessary. Results: A total of 11 patients (mean age 12.2 years (9-14)) were included, and the mean follow-up period was 36.5 (21-54) months. Most fractures were caused by direct or indirect trauma (n = 8), and others (n = 3) underwent PPF without obvious cause. The first type of treatment was performed on four patients, and the second type was performed on seven patients. The mean Musculoskeletal Tumor Society (MSTS) score was 20 (17-23). All patients recovered from the complication, and limb preservation could be achieved. Conclusion: PPF is a big challenge for musculoskeletal oncologists, particularly in younger patients. Additionally, PPF poses a challenge for orthopedic surgeons, as limb preservation should be an important goal. Hence, internal fixation with plates and endoprosthetic replacement are optional treatment strategies based on fracture type and patient needs.

Cancer Cell Membrane-Coated Nanoparticle Co-loaded with Photosensitizer and Toll-like Receptor 7 Agonist for the Enhancement of Combined Tumor Immunotherapy.

Tumor immunotherapy has shown considerable therapeutic potential in the past few years, but the clinical response rate of immunotherapy is less than 20%. Encountering the high heterogeneity of tumors, it will be a general trend to apply combined therapy for cancer treatment. Photodynamic therapy (PDT) transiently kills tumor cells by producing reactive oxygen species (ROS), while residual tumor cells are prone to metastasis, leading to tumor recurrence. In combination with tumor immunotherapy, it is hoped to awaken the host immune system and eradicate residual tumor cells. Herein, cancer cell membrane-coated nanoparticles as a platform to combine PDT, TLR7 agonist, and tumor antigen for the enhancement of tumor therapeutic efficacy are designed. The final biomimetic nanoparticles (CCMV/LTNPs) can specifically kill tumor cells through PDT, while strong host antitumor immune responses are elicited to eliminate residue tumor cells under the help of immune adjuvant and tumor antigen from the cancer cell membrane. In summary, a photoimmunotherapy strategy is designed that synergistically enhances the tumor therapeutic effects by killing tumor cells through PDT and activating host antitumor immune responses through the co-delivery of adjuvant and tumor antigen, which may offer a promising strategy for clinical immunotherapy in the future.

Association of circulating tumor cells and IMP3 expression with metastasis of osteosarcoma.

Background: Circulating tumor cells (CTCs) have been identified as a prognostic biomarker of tumors such as breast cancer and nasopharyngeal carcinoma, because they are obtained through a simple and noninvasive blood draw or liquid biopsy, but its clinical significance in osteosarcoma is still unclear. In this study, we analyzed the relationship between CTCs and clinicopathological features and discussed whether CTCs could be used as a biomarker for metastasis in osteosarcoma. Methods: We enrolled 50 osteosarcoma patients with Enneking Stage IIB and Stage III and detected CTCs in 5 ml of peripheral blood samples collected from patients using the Canpatrol® CTC detection platform. Subsequently, multiplex RNA in situ hybridization (RNA-ISH) based on various molecular markers was performed to identify and classify CTCs. The relationships between clinical pathological features and CTC counts, subtypes (epithelial type, E type; hybrid epithelial/mesenchymal type, H type; mesenchymal type, M type), and insulin-like growth factor mRNA-binding protein 3 (IMP3) expression in CTCs were analyzed. Results: CTCs were detected in 86% (43/50) of the osteosarcoma patients. The CTC counts, especially the total CTCs and H-type CTCs, signifcantly differed between Enneking Stage IIB and Stage III patients (P < 0.05). No significant differences were observed between the CTC count or type and other clinicopathological features (P > 0.05). There were significant differences in the expression of IMP3 in different types of CTCs, and the IMP3 positive rates in E/H/M type CTCs were 38.4, 65.6, and 62.0%, respectively (P < 0.001). Receiver operating characteristic (ROC) curve analysis showed that IMP3-positive CTC count had the best performance for diagnostic metastasis, with the largest area under the curve of 0.873 and cutoff value of four cells/5ml blood (sensitivity = 87.5%; specificity = 82.4%). Serial CTC monitoring in one patient suggested that total CTCs and H-type CTCs were associated with disease progression. Conclusion: This study demonstrates that the CTCs, especially the IMP3-positive CTCs and H/M-type CTCs, are related to the metastasis of osteosarcoma.

HMGB1 Positive Feedback Loop Between Cancer Cells and Tumor-Associated Macrophages Promotes Osteosarcoma Migration and Invasion.

Numerous studies have demonstrated the key roles of tumor-associated macrophages (TAMs) in osteosarcoma metastasis. Higher levels of high mobility group box 1 (HMGB1) promote osteosarcoma progression. However, whether HMGB1 is involved in the polarization of M2 macrophages into M1 macrophages in osteosarcoma remains largely unknown. Here, HMGB1 and CD206 mRNA expression levels were measured by a quantitative reverse transcription-polymerase chain reaction in osteosarcoma tissues and cells. HMGB1 and receptor for advanced glycation end products (RAGE) protein expression levels were measured by western blotting. Osteosarcoma migration was measured using transwell and wound-healing assays, while a transwell assay determined osteosarcoma invasion. Macrophage subtypes were detected using flow cytometry. HMGB1 expression levels were aberrantly enhanced in osteosarcoma tissues compared with normal tissues and were positively correlated with AJCC III and IV stages, lymph node metastasis, and distant metastasis. Silencing HMGB1 inhibited the migration, invasion, and epithelial-mesenchymal transition (EMT) of osteosarcoma cells. Furthermore, reduced HMGB1 expression levels in conditioned media derived from osteosarcoma cells induced the polarization of M2 TAMs to M1 TAMs. In addition, silencing HMGB1 inhibited the liver and lung metastasis of tumors and reduced the expression levels of HMGB1, CD163, and CD206 in vivo. HMGB1 was found to regulate macrophage polarization through RAGE. Polarized M2 macrophages induced osteosarcoma migration and invasion, activating HMGB1 expression in osteosarcoma cells to form a positive feedback loop. In conclusion, HMGB1 and M2 macrophages enhanced osteosarcoma migration, invasion, and EMT through positive feedback regulation. These findings reveal the significance of tumor cell and TAM interactions in the metastatic microenvironment.

Effect of Dietary Supplemental Zinc on Laying Performance, Egg Quality, and Plasma Hormone Levels of Breeding Pigeons.

This study aimed to evaluate the dietary zinc requirement of parental pigeons for better laying and reproductive performance, egg quality, sex hormones, and mineral content in eggs. A total of 160 pairs of healthy American Silver King pigeons were randomly assigned to five treatments of eight replicate cages each with four pairs of birds per cage, and fed a basal diet without zinc supplementation or the basal diet supplemented with 30, 60, 90, and 120 mg of zinc/kg (ZnSO4·7H2O). The experiment lasted for 45 days, including two laying cycles. Results indicated the egg production rate (P = 0.081), egg shape index (P = 0.038), egg eggshell percentage (P = 0.070), and zinc and calcium contents (P < 0.01) tended to be affected or significantly affected by zinc addition. They increased quadratically with dietary zinc levels (P < 0.05). Besides, shell thickness (P = 0.069), plasma testosterone (P = 0.008), LH, and carbonic anhydrase contents (P < 0.05) tended to be affected or significantly affected by zinc addition. They increased linearly as dietary zinc level increased (P < 0.05). Compared with the control, 60 mg/kg zinc addition increased egg production rate, egg shape index, zinc and calcium contents in eggshell, and plasma testosterone concentration in pigeons (P < 0.05), and tended to increase the eggshell percentage (P = 0.07). Besides, supplemental 120 mg/kg zinc had higher shell thickness and LH content than control (P < 0.05), but had no difference with 60 mg/kg zinc addition. In conclusion, the supplementation of zinc at the level of 60 mg/kg to basal diet improved laying performance by increasing eggshell quality and sex hormone levels of breeding pigeons.

En bloc resection of huge primary tumors with epidural involvement in the mobile spine using the "rotation-reversion" technique: Feasibility, safety, and clinical outcome of 11 cases.

Background: En bloc resection of spinal tumors provides better local control and survival outcomes than intralesional resection. Safe margins during en bloc resection of primary spinal tumors with epidural involvement are required for improved outcomes. The present study describes a "rotation-reversion" technique that has been used for en bloc resection of huge primary tumors in the mobile spine with epidural involvement and reported the clinical outcomes in these patients. Methods: All patients with primary spinal tumors who were treated with the rotation-reversion technique at our institution between 2015 and 2021 were evaluated retrospectively. Of the patients identified, those with both huge extraosseous soft-tissue masses and epidural involvement were selected for a case review. Clinical and radiological characteristics, pathologic findings, operative procedures, complications, and oncological and functional outcomes of these patients were reviewed. Results: Of the 86 patients identified with primary spinal tumors who underwent en bloc resection using the rotation-reversion technique between 2015 and 2021, 11 had huge extraosseous soft-tissue masses with epidural involvement in the mobile spine. The average maximum size of these 11 tumors was 8.1 × 7.5 × 9.7 cm. Median follow-up time was 28.1 months, mean operation time was 849.1 min (range 465-1,340 min), and mean blood loss was 6,972.7 ml (range 2,500-17,700 ml), with 10 (91%) of the 11 patients experiencing perioperative complications. The negative margin rate was 91%, with only one patient (9%) experiencing local recurrence. Ten patients were able to walk normally or with a crutch at the last follow-up, whereas one was completely paralyzed preoperatively. Conclusion: The rotation-reversion technique is an effective procedure for the en bloc resection of huge primary spinal tumors, with the extension of invasion in selected patients including not only the vertebral body but also the pedicle and part of the posterior arch.

Image-guided percutaneous biopsy and pathological diagnosis in atypical tuberculous spondylitis: a case series and clinical outcomes.

Background: Tuberculous spondylitis can be difficult to distinguish from alternative spinal pathologies such as malignancy, particularly if the imaging features are not typical. Biopsy and histopathological analysis are facilitative to the early and accurate diagnosis of atypical tuberculous spondylitis and the clinical management. The purpose of this study is to describe some of the atypical imaging features of tuberculous spondylitis diagnosed by image-guided percutaneous biopsy, as well as associated treatment outcomes. Methods: We performed a retrospective analysis of all patients diagnosed with tuberculous spondylitis after image-guided percutaneous biopsy at The Third Affiliated Hospital of Southern Medical University between 2013 and 2020. Of the patients identified, those with atypical imaging features were selected for case review. All patients were given anti-tuberculous medication treatment with or without surgery. The imaging features, histological and microbiological results, and clinical presentations and outcomes were evaluated. Neurological function was evaluated according to the Frankel grading system. The clinical outcomes were evaluated by Visual Analogic Scale (VAS) scores for pain, imaging [X-ray, computed tomography (CT), and magnetic resonance imaging (MRI)] results, and laboratory examinations. Comparison of VAS scores was made by Student t-test. Results: Of the 102 patients identified with tuberculous spondylitis between 2013 and 2020, eight patients (two females and six males) with a mean age of 41.6 years (range, 18-61 years) demonstrated atypical imaging findings, including central vertebral body lesion, multiple skip vertebral lesions, extradural mass lesion and anterior subperiosteal lesion. All eight patients received anti-tuberculous medication treatment, and six underwent surgery. One patient developed a pleural effusion after debridement of the thoracic lesion. The mean follow-up period was 16.2 months (6-37 months). The VAS scores before treatment and at the final follow-up showed significant differences (7.25±1.49 and 0.0±0.0, respectively, P<0.01). Improved neurological function were observed in all patients. Solid fusion and osteogenic osteosclerosis were observed at the final follow-up, and no recurrence was observed in any cases. Conclusions: All eight patients had a good prognosis. Image-guided biopsy and histopathological analysis are helpful for the early diagnosis of tuberculous spondylitis, especially when imaging features are not typical for this condition.

Small molecule SMU-CX24 targeting toll-like receptor 3 counteracts inflammation: A novel approach to atherosclerosis therapy.

Toll-like receptor 3 (TLR3), as an important pattern recognition receptor (PRR), dominates the innate and adaptive immunity regulating many acute and chronic inflammatory diseases. Atherosclerosis is proved as an inflammatory disease, and inflammatory events involved in the entire process of initiation and deterioration. However, the contribution of TLR3 to atherosclerosis remains unclear. Herein, we identified the clinical relevance of TLR3 upregulation and disease processes in human atherosclerosis. Besides, activation of TLR3 also directly led to significant expression of atherogenic chemokines and adhesion molecules. Conversely, silencing TLR3 inhibited the uptake of oxLDL by macrophages and significantly reduced foam cell formation. Given the aberrance in TLR3 functions on atherosclerosis progression, we hypothesized that TLR3 could serve as novel target for clinical atherosclerosis therapy. Therefore, we developed the novel ellipticine derivative SMU-CX24, which specifically inhibited TLR3 (IC50 = 18.87 ± 2.21 nmol/L). In vivo, atherosclerotic burden was alleviated in Western diet fed ApoE-/- mice in response to SMU-CX24 treatment, accompanying notable reductions in TLR3 expression and inflammation infiltration within atherosclerotic lesion. Thus, for the first time, we revealed that pharmacological downregulation of TLR3 with specific inhibitor regenerated inflammatory environment to counteract atherosclerosis progression, thereby proposing a new strategy and probe for atherosclerosis therapy.

Influence of dietary phosphorus concentrations on the performance of rearing pigeons (Columba livia), and bone properties of squabs.

The objective of this study was to investigate the effects of dietary P levels on the performance of rearing pigeons, and bone characteristics of squabs from 7 to 21 d of age. A total of 192 pairs of adult Silver King pigeons (40 wk of age) were used. The pigeons were randomly allocated to one of 4 treatment groups, each consisting of eight replicates of 6 pigeon pairs per replicate. Dietary treatments included the basal diet (containing 0.3% of P), the basal diet supplemented with 0.2, 0.4, or 0.8% inorganic P. And the dietary Ca content was kept at 1.40% across all treatments. The experimental diets were fed to parent pigeons as corn-soybean complete pellet feed, and squabs fed with crop milk secreted by parent pigeons. Pigeons in the group of 0.4% supplemental non-phytate phosphorus (NPP) had shorter (P = 0.045) oviposition interval than those in the control group and group of 0.8% NPP. When the diet was supplemented with 0.8% of NPP, the least average egg weight was observed (P = 0.006). Female breeding birds had much higher (P < 0.01) Ca, P, and ALP in serum than male ones. At 7-d of age, dietary P supplementation influenced P and Ca content in tibia ash of squabs (P < 0.05). The tibia ash Ca content in the group of 0.2% NPP was the highest among the treatments (P = 0.007). At d 21 of age, both the birds in the group of 0.4 and 0.8% NPP had higher tibia breaking strength (P < 0.01) and tibia ash contents (P < 0.001) compared to the ones in the control group. In conclusion, the P deficiency in the diet of parent pigeons could cause poor bone mineralization of squabs, especially impaired the bone-breaking strength and bone ash content. The 0.8% of NPP supplementation in the diet has a positive influence on mineralization of squabs although production depression was observed. Both P and Ca metabolism of female breeding birds were more active than male ones at earlier time points of rearing period. The desirable supplemental NPP level in diet for breeding pigeon was 0.4% according to the performance data in the present trial. The recommended Ca: P ratio for pigeons, which was different from the optimum value for broilers, needs to be studied in the future.

Protective Effects of Zinc on Salmonella Invasion, Intestinal Morphology and Immune Response of Young Pigeons Infected with Salmonella enterica Serovar Typhimurium.

The study aimed to determine the effects of orally supplemental zinc on body weight, Salmonella invasion, serum IgA, intestinal histomorphology, and immune response of Salmonella enterica serovar Typhimurium (S. typhimurium)-challenged young pigeons. A total of 72 healthy White King pigeons (25 days old) with similar weight were randomly assigned to 3 treatments with six replicate cages. The 3 treatments were unchallenged, S. typhimurium-challenged, and S. typhimurium-challenged orally supplemented with 1 mg zinc per bird. Salmonella infection decreased (P < 0.05) the body weight, the bursa index, the serum IgA content, and the villus height/crypt depth ratio in the ileum, but increased the neutrophil proportion (P < 0.001) and the mRNA expressions of IL-1ß and IL-8 in the jejunum (P < 0.05). Orally supplemental zinc reduced (P = 0.007) the bacterial load in the liver and improved (P < 0.05) the body weight, the bursa index, the serum IgA content, the villus height/crypt depth ratio, and the NOD-like receptor family pyrin domain containing 3 (NLRP3) protein expression, as well as tended to increase (P = 0.064) the protein abundance of caspase-1 of the jejunum, but did not alleviate the high level of neutrophil proportion and IL-1ß mRNA expression of the jejunum (P > 0.05). The results indicated that oral zinc supplementation improved the intestinal mucosal morphology and enhanced the immune response, as well as activated caspase-1-dependent cell pyroptosis pathways in the jejunal epithelium, thereby restricting Salmonella invasion of the challenged young pigeons.

Effects of Yucca schidigera extract on serum biochemical parameters, humoral immune response, and intestinal health in young pigeons.

Introduction: It is of great importance to find antibiotic alternatives that can improve poultry performance and enhance immunity. Plant-derived extracts and their concentrates are natural bioactive compounds that are widely and effectively applied as the antibiotic alternatives in animal industries. This study was conducted to investigate the effects of Yucca schidigera extract (YSE) on growth performance, serum biochemical parameters, immune function, intestinal morphology, and microbiota diversity of young pigeons. Methods: A total of 120 healthy White King pigeons (28 days old) with similar weight were randomly assigned to 4 treatments with six replicate cages. Each of the pigeons from 4 treatments was orally administrated with 0 (control), 5, 10, and 15 mg YSE per day, respectively. Results: The results showed that orally supplemental YSE had no significant effects (P > 0.05) on the growth performance and immune organ index of pigeons. The serum total protein and IgM contents in the 10 mg YSE group were significantly higher (P < 0.05) than those in the control group. Supplemental 10 and 15 mg YSE significantly lowered the level of serum total cholesterol (P < 0.05) and increased (P < 0.05) the villi height in the jejunum compared with the control group. Supplemental 5 and 10 mg YSE significantly decreased (P < 0.05) the level of serum alanine aminotransferase and the crypt depth in the ileum compared with the control group. The beta diversity showed a distinct difference in the ileum microbial composition between the control and the 10 mg YES group. YSE supplementation enriched the bacterial genera Sulfurospirillum, Solobacterium, Desulfovibrio, Desulfobulbus, Lactococcus, Parabacteroides, Acidaminococcus, Acetobacter, and Streptococcus. Additionally, Enterococcus genus showed a significantly negative correlation with serum alanine aminotransferase (R = -0.618, P = 0.043). Actinomyces genus showed a significantly negative correlation with cholesterol (R = -0.633, P = 0.036). Turicibacter genus showed a significantly positive correlation with villi height in the jejunum (R = 0.751, P = 0.008). Discussion: In conclusion, orally supplemental YSE could improve serum biochemistry, immunoglobulin contents, and intestinal morphology by regulating the composition of microbial community in the ileum of young pigeons.

Exploration of Proteomics Analysis of Crop Milk in Pigeons (Columba livia) during the Lactation Period.

Pigeon milk is a curdlike substance separated from the mature crop epithelium of breeders, associated with the rapid growth and development of squabs. The aim of this study was to investigate in detail the variations in the content of several important ingredients in crop milk. In this study, we utilized proteomic techniques to investigate the composition and changing pattern of crop milk protein of squabs on days 1 (D1), 3 (D3), and 7 (D7). Our results indicated that the crude protein contents in crop milk decreased with age, and they were up to 50% during the first 3 days. The proteomic data showed that a total of 2558 proteins were identified in all samples from three stages, and the top 15% crop milk proteins were ribosomal protein, keratin, peroxiredoxin, annexin, heat shock protein, and eukaryotic translation protein based on liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis and normalized spectral abundance factors (NSAFs) calculation. Furthermore, the compositions of crop milk protein between D1 and D3 were quite similar [51 differentially expressed proteins (DEPs)], while great proteomic differences were observed between D1/D3 and D7 (more than 240 DEPs). Additionally, gene ontology (GO) enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis showed that up-regulated DEPs mainly participate in immune response, while down-regulated DEPs were involved in cell differentiation and development as well as tRNA aminoacylation biosynthesis. In conclusion, DEPs were mainly related to protein synthesis, immunity, and antioxidation, which provided effective information for the development of artificial squab milk products in the future.

Indigenous Bacteria Have High Potential for Promoting Salix integra Thunb. Remediation of Lead-Contaminated Soil by Adjusting Soil Properties.

Salix integra Thunb., a fast-growing woody plant species, has been used for phytoremediation in recent years. However, little knowledge is available regarding indigenous soil microbial communities associated with the S. integra phytoextraction process. In this study, we used an Illumina MiSeq platform to explore the indigenous microbial composition after planting S. integra at different lead (Pb) contamination levels: no Pb, low Pb treatment (Pb 500 mg kg-1), and high Pb treatment (Pb 1500 mg kg-1). At the same time, the soil properties and their relationship with the bacterial communities were analyzed. The results showed that Pb concentration was highest in the root reaching at 3159.92 ± 138.98 mg kg-1 under the high Pb treatment. Planting S. integra decreased the total Pb concentration by 84.61 and 29.24 mg kg-1, and increased the acid-soluble Pb proportion by 1.0 and 0.75% in the rhizosphere and bulk soil under the low Pb treatment compared with unplanted soil, respectively. However, it occurred only in the rhizosphere soil under the high Pb treatment. The bacterial community structure and microbial metabolism were related to Pb contamination levels and planting of S. integra, while the bacterial diversity was only affected by Pb contamination levels. The dominant microbial species were similar, but their relative abundance shifted in different treatments. Most of the specific bacterial assemblages whose relative abundances were promoted by root activity and/or Pb contamination were suitable for use in plant-microbial combination remediation, especially many genera coming from Proteobacteria. Redundancy analysis (RDA) showed available nitrogen and pH having a significant effect on the bacteria relating to phytoremediation. The results indicated that indigenous bacteria have great potential in the application of combined S. integra-microbe remediation of lead-contaminated soil by adjusting soil properties.

Experimental investigation on rockburst behavior of the rock-coal-bolt specimen under different stress conditions.

Coal and rock burst are one of the main dynamic disasters that affect coal mine production. In this paper, the burst structural model of the rock-coal-bolt (RCB) system and the burst tendency criterion are established on the background of deep thin coal seam mining. Uniaxial and triaxial mechanical tests under different stress states are carried out on RCB specimens with different angles. Combined with thermal imaging, the mechanical behavior of the inclined RCB specimen under uniaxial loading is discussed. The results show that the burst tendency of the RCB specimen increases with the angle. The stress-strain curves of some uniaxial and triaxial test specimens show two or more peaks, and the thermal imaging evolutionary process shows that the cracks of the coal and rock develop from shear to tension shear cracks. There is a further development of fracture and energy accumulation between the first and second peaks in the stress-strain curve of the specimen. Therefore, the failure degree of the second peak of the specimen may be stronger than that of the first peak. Additionally, the established stiffness coefficient and burst energy index can better describe the burst tendency of the RCB specimen under different stress states. The results show that the burst tendency of the RCB specimen under the triaxial test is much higher than that of the uniaxial test. In other words, it also explains that the essence of the burst failure of the surrounding rock in the roadway is the initial instability induced by the inside surrounding rock in the roadway. Moreover, the burst tendency is the largest when the rock and coal combination angle is 15°, and the burst damage range may also be increased by the failure of internal coal and rock mass.

Application of a novel porous tantalum implant in rabbit anterior lumbar spine fusion model: in vitro and in vivo experiments.

BACKGROUND: Some porous materials have been developed to enhance biologic fusion of the implants to bone in spine fusion surgeries. However, there are several inherent limitations. In this study, a novel biomedical porous tantalum was applied to in vitro and in vivo experiments to test its biocompatibility and osteocompatibility. METHODS: Bone marrow-derived mesenchymal stem cells (BMSCs) were cultured on porous tantalum implant. Scanning electron microscope (SEM) and Cell Counting Kit-8 assay were used to evaluate the cell toxicity and biocompatibility. Twenty-four rabbits were performed discectomy only (control group), discectomy with autologous bone implanted (autograft group), and discectomy with porous tantalum implanted (tantalum group) at 3 levels: L3-L4, L4-L5, and L5-L6 in random order. All the 24 rabbits were randomly sacrificed at the different post-operative times (2, 4, 6, and 12 months; n = 6 at each time point). Histologic examination and micro-computed tomography scans were done to evaluate the fusion process. Comparison of fusion index scores between groups was analyzed using one-way analysis of variance. Other comparisons of numerical variables between groups were made by Student t test. RESULTS: All rabbits survived and recovered without any symptoms of nerve injury. Radiographic fusion index scores at 12 months post-operatively between autograft and tantalum groups showed no significant difference (2.89 ±â€Š0.32 vs. 2.83 ±â€Š0.38, F = 244.60, P = 0.709). Cell Counting Kit-8 assay showed no significant difference of absorbance values between the leaching liquor group and control group (1.25 ±â€Š0.06 vs. 1.23 ±â€Š0.04, t = -0.644, P = 0.545), which indicated the BMSC proliferation without toxicity. SEM images showed that these cells had irregular shapes with long spindles adhered to the surface of tantalum implant. No implant degradation, wear debris, or osteolysis was observed. Histologic results showed solid fusion in the porous tantalum and autologous bone implanted intervertebral spaces. CONCLUSION: This novel porous tantalum implant showed a good biocompatibility and osteocompatibility, which could be a valid biomaterial for interbody fusion cages.

Evidence that miR-146a attenuates aging- and trauma-induced osteoarthritis by inhibiting Notch1, IL-6, and IL-1 mediated catabolism.

Primary osteoarthritis (OA) is associated with aging, while post-traumatic OA (PTOA) is associated with mechanical injury and inflammation. It is not clear whether the two types of osteoarthritis share common mechanisms. We found that miR-146a, a microRNA-associated with inflammation, is activated by cyclic load in the physiological range but suppressed by mechanical overload in human articular chondrocytes. Furthermore, miR-146a expression is decreased in the OA lesions of human articular cartilage. To understand the role of miR-146a in osteoarthritis, we systemically characterized mice in which miR-146a is either deficient in whole body or overexpressed in chondrogenic cells specifically. miR-146a-deficient mice develop early onset of OA characterized by cartilage degeneration, synovitis, and osteophytes. Conversely, miR-146a chondrogenic overexpressing mice are resistant to aging-associated OA. Loss of miR-146a exacerbates articular cartilage degeneration during PTOA, while chondrogenic overexpression of miR-146a inhibits PTOA. Thus, miR-146a inhibits both OA and PTOA in mice, suggesting a common protective mechanism initiated by miR-146a. miR-146a suppresses IL-1ß of catabolic factors, and we provide evidence that miR-146a directly inhibits Notch1 expression. Therefore, such inhibition of Notch1 may explain suppression of inflammatory mediators by miR-146a. Chondrogenic overexpression of miR-146a or intra-articular administration of a Notch1 inhibitor alleviates IL-1ß-induced catabolism and rescues joint degeneration in miR-146a-deficient mice, suggesting that miR-146a is sufficient to protect OA pathogenesis by inhibiting Notch signaling in the joint. Thus, miR-146a may be used to counter both aging-associated OA and mechanical injury-/inflammation-induced PTOA.

Global Reliability Sensitivity Analysis Based on Maximum Entropy and 2-Layer Polynomial Chaos Expansion.

To optimize contributions of uncertain input variables on the statistical parameter of given model, e.g., reliability, global reliability sensitivity analysis (GRSA) provides an appropriate tool to quantify the effects. However, it may be difficult to calculate global reliability sensitivity indices compared with the traditional global sensitivity indices of model output, because statistical parameters are more difficult to obtain, Monte Carlo simulation (MCS)-related methods seem to be the only ways for GRSA but they are usually computationally demanding. This paper presents a new non-MCS calculation to evaluate global reliability sensitivity indices. This method proposes: (i) a 2-layer polynomial chaos expansion (PCE) framework to solve the global reliability sensitivity indices; and (ii) an efficient method to build a surrogate model of the statistical parameter using the maximum entropy (ME) method with the moments provided by PCE. This method has a dramatically reduced computational cost compared with traditional approaches. Two examples are introduced to demonstrate the efficiency and accuracy of the proposed method. It also suggests that the important ranking of model output and associated failure probability may be different, which could help improve the understanding of the given model in further optimization design.

Overendocytosis of superparamagnetic iron oxide particles increases apoptosis and triggers autophagic cell death in human osteosarcoma cell under a spinning magnetic field.

The toxicity of superparamagnetic iron oxide nanoparticles (SPIONs) is still a vital topic of debate and the mechanisms remain unclear. In the present study, overdose SPIONs could induce osteosarcoma cell death and the effects were exaggerated when combined with spinning magnetic field (SMF). In the combination group, mitochondrial transmembrane potential decrease more obviously and reactive oxygen species (ROS) was found to generate much higher in line with that of the apoptosis ratio. Meantime, amount of autophagy was induced. Inhibiting the autophagy generation by 3-methyladenine (3-MA) increase cell viability but decrease the caspase 3/7 and caspase 8 activities in combination groups, and inhibiting apoptosis took the same effect. In the end, the SPIONs effects on xenograft mice was examed by intratumoral injection. The result showed that the combination group could greatly decrease the tumor volume and prolong the lifespan of mice. In sum, the result indicated that overdose SPIONs induced ROS generation, and excessive ROS induced by combination of SPIONs and SMF contribute to autophagy formation, which play a apoptosis-promoting role that formed as a platform to recruits initiate the caspase activities.

FKBP14 overexpression contributes to osteosarcoma carcinogenesis and indicates poor survival outcome.

The FK506-binding protein 14 (FKBP14) is a subfamily of immunophilins, has been implicated in various biochemical processes. However, its effects on the primary malignant bone tumor, osteosarcoma, are unclear. Here, we reported that FKBP14 may be an oncogene as it overexpressed in osteosarcoma tissues and cell lines, and FKBP14 expression was correlated with metastases, recurrence, tumor maximum diameter and poor survival time. FKBP14 was associated with the biological pathways including cell cycle, apoptosis and metastasis. Furthermore, we detected FKBP14 knockdown induced cell cycle arrest, apoptosis, invasion and adhesion in vitro. FKBP14 knockdown decreased the protein levels of PCNA, CDK1 and CCNB1 that promotes cell cycle, increased Bax, caspase-3 and caspase-7 protein involved in promoting cell apoptosis, and increased KIF4A expression as well as decreased SMC4 and TMEM33 proteins that contribute to cell invasion and adhesion. In addition, FKBP14 knockdown also caused a significant inhibition in tumor growth in vivo. Then, we found that the protein RhoA was identified as a binding partner of FKBP14. Taken together, FKBP14 may act as an oncogene in osteosarcoma via suppressing apoptosis and promoting invasion and adhesion in osteosarcoma carcinogenesis. FKBP14 may be a prognostic factor and potential target for osteosarcoma treatment.

Effects of UCP4 on the Proliferation and Apoptosis of Chondrocytes: Its Possible Involvement and Regulation in Osteoarthritis.

Reactive oxygen species (ROS)-induced chondrocytes apoptosis plays a key role in osteoarthritis (OA) pathogenesis. Uncoupling protein 4 (UCP4) can protect cells against oxidative stress via reducing ROS production and cell apoptosis. Here, silencing of UCP4 in primary chondrocytes significantly inhibited cell survival, but induced ROS production and cell apoptosis. UCP4 mRNA of cartilage tissues was decreased in osteoarthritis patients, which was negatively correlated with synovial fluid (SF) leptin concentration. Moreover, leptin treatment (5, 10 and 20 ng/ml) of primary cultured chondrocytes significantly decreased mRNA and protein levels of UCP4, but increased ROS production and cell apoptosis in a dose-dependent manner. The effects of leptin treatment (20 ng/ml) on chondrocytes was partially reversed by ectopic expression of UCP4. More importantly, intraarticularly injection of UCP4 adenovirus remarkably alleviate OA progression and cell apoptosis in a rat OA model induced by anterior cruciate ligament transection (ACLT). In conclusion, UCP4, whose expression was suppressed by leptin, may be involved in the ROS production and apoptosis of chondrocytes, thus contributing to the OA pathogenesis.