Experimental Study of Shear Performance of High-Strength Concrete Deep Beams with Longitudinal Reinforcement with Anchor Plate.

As a transfer member at the discontinuous place of vertical load, the deep beam has a complex stress mechanism and many influencing factors, such as compressive strength of concrete, shear span ratio, and reinforcement ratio. At the same time, the stress analysis principle of traditional shallow beams is no longer applicable to the design and calculation of deep-beam structure. The main purpose of this paper was to use the strut-and-tie model to analyze its stress mechanism, and to verify the applicability of the model. Nine high-strength concrete deep-beam specimens with longitudinal reinforcement with an anchor plate of the same size were tested by two-point concentrated loading method. The effects of shear span ratio (0.3, 0.6, and 0.9), longitudinal reinforcement ratio (0.67%, 1.05%, and 1.25%), horizontal reinforcement ratio (0.33%, 0.45%, and 0.50%), and stirrup reinforcement ratio (0.25%, 0.33%, and 0.50%) on the failure mode, deflection curve, characteristic load, crack width, steel bar, and concrete strain of the specimens were analyzed. The results showed that the failure mode of deep-beam specimens was diagonal compression failure. The normal section cracking load was about 15 to 20% of the ultimate load, and the inclined section cracking load was about 30~40% of the ultimate load. The shear span ratio increased from 0.3 to 0.9, and the bearing capacity decreased by 32.9%. When the longitudinal reinforcement ratio increased from 0.67% to 1.25%, the ultimate load increased by 42.6%. The shear span ratio and longitudinal reinforcement ratio have a significant effect on the bearing capacity of the high-strength concrete deep beams with longitudinal reinforcement with an anchor plate. The shear capacity of nine high-strength concrete deep-beam specimens with longitudinal reinforcement with an anchor plate was calculated by national standards, and the results were compared with the calculation results of the Tan-Tang model, the Tan-Cheng model, SSTM, and SSSTM. The analysis showed that the softened strut-and-tie model takes into account the softening effect of compressive concrete, and is a more accurate mechanical model, which can be applied to predict the shear capacity of high-strength concrete deep-beam members with longitudinal reinforcement with an anchor plate.

Analysis of Shear Model for Steel-Fiber-Reinforced High-Strength Concrete Corbels with Welded-Anchorage Longitudinal Reinforcement.

According to the shear capacity test results of six steel-fiber-reinforced high-strength concrete (SFHSC) corbels with welded-anchorage longitudinal reinforcement under concentrated load, the effects of shear span ratio and steel fiber volume fraction on the failure mode, cracking load and ultimate load of corbel specimens were analyzed. On the basis of experimental research, the shear transfer mechanism of corbel structure was discussed. Then, a modified softened strut-and-tie model (MSSTM), composed of the diagonal and horizontal mechanisms, was proposed, for steel-fiber-reinforced high-strength concrete corbels. The contributions of concrete, steel fiber and horizontal stirrups to the shear bearing capacity of the corbels were clarified. A calculation method for the shear bearing capacity of steel-fiber-reinforced high-strength concrete corbels was established and was simplified on this basis. The calculation results of the model were compared with the test values and calculation results of the GB50010-2010 code, the ACI318-19 code, the EN 1992-1-1 code and the CSA A23.3-19 code. The results showed that the concrete corbel with small shear span ratio mainly has two typical failure modes: shear failure and diagonal compression failure. With the increase in shear span ratio, the shear capacity of corbels decreases. Steel fiber can improve the ductility of a reinforced concrete corbel, but has little effect on the failure mode of the diagonal section. The calculated values of the national codes were lower than the experimental values, and the results were conservative. The theoretical calculation values of the shear capacity calculation model of the corbels were close to the experimental results. In addition, the model has a clear mechanical concept considering the tensile properties of steel-fiber-reinforced high-strength concrete and the influence of horizontal stirrups, which can reasonably reflect the shear transfer mechanism of corbels.

Low miR-182-5p Expressing Extracellular Vesicles Derived From Human Bone Marrow Stromal Cells of Subjects With Steroid-Induced Osteonecrosis of the Femoral Head Aggravate Disease Progression.

Steroid-induced osteonecrosis of the femoral head (SONFH) is a refractory, progressive disease. However, the underlying mechanisms that aggravate femoral head necrosis remain unclear. Extracellular vesicles (EVs) act as molecular carriers in intercellular communication. We hypothesize that EVs derived from human (h) bone marrow stromal cells (BMSC) resident in SONFH lesion areas promote the pathogenesis of SONFH. In the present study, we determined the modulatory effects of SONFH-hBMSCs-derived EVs on the pathogenesis of SONFH in vitro and in vivo. We found that the expression of hsa-miR-182-5p was downregulated in SONFH-hBMSCs and EVs isolated from those hBMSCs. After tail vein injection, EVs isolated from hBMSCs transfected with hsa-miR-182-5p inhibitor aggravated femoral head necrosis in the SONFH mouse model. We conclude that miR-182-5p regulates bone turnover in the SONFH mouse model via targeting MYD88 and subsequent upregulation of RUNX2 expression. We further assume that EVs derived from hBMSCs resident in SONFH lesion areas aggravate femoral head necrosis by downregulating miR-182-5p secreted from hBMSC located outside these lesions. We suggest that miR-182-5p could provide a novel target for future therapeutic approaches to treat or prevent SONFH. © 2023 American Society for Bone and Mineral Research (ASBMR).

Experimental Investigation on Shear Capacity of Steel-Fiber-Reinforced High-Strength Concrete Corbels.

As short cantilever members, corbels are mainly used to transfer eccentric loads to columns. Because of the discontinuity of load and geometric structure, corbels cannot be analyzed and designed using the method based on beam theory. Nine steel-fiber-reinforced high-strength concrete (SFRHSC) corbels were tested. The width of the corbels was 200 mm, the cross-section height of the corbel column was 450 mm, and the cantilever end height was 200 mm. The shear span/depth ratios considered were 0.2, 0.3, and 0.4; the longitudinal reinforcement ratios were 0.55%, 0.75%, and 0.98%; the stirrup reinforcement ratios were 0.39%, 0.52%, and 0.785%; and the steel fiber volume ratios were 0, 0.75%, and 1.5%. According to the test results, this paper discusses the failure process and failure mode of corbel specimens with a small shear span/depth ratio and analyzes the effects of variables such as shear span/depth ratio, longitudinal reinforcement ratio, stirrup reinforcement ratio, and steel fiber volume content on the shear capacity of corbels. The shear capacity of corbels is significantly affected by the shear span/depth ratio, followed by the longitudinal reinforcement ratio and the stirrup reinforcement ratio. Moreover, it is shown that steel fibers have little impact on the failure mode and ultimate load of corbels, but can enhance the crack resistance of corbels. In addition, the bearing capacities of these corbels were calculated by Chinese code GB 50010-2010 and further compared with ACI 318-19 code, EN 1992-1-1:2004 code, and CSA A23.3-19 code, which adopt the strut-and-tie model. The results indicate that the calculation results by the empirical formula in the Chinese code are close to the corresponding test results, while the calculation method based on the strut-and-tie model of a clear mechanical concept yields conservative results, and hence the related parameter values must be further modified.

Incorporating paraffin@SiO2 nanocapsules with abundant surface hydroxyl groups into polydimethylsiloxane to develop composites with enhanced interfacial heat conductance for chip heat dissipation.

Incorporating phase change capsules into polymeric matrices is an effective approach for developing flexible composites with both heat storage capacity and good thermal reliability, while the interfacial heat conductance between the capsules and the matrix has seldom been considered. Herein, paraffin@SiO2 nanocapsules synthesized by an interfacial polycondensation process using a basic catalyst were incorporated into a polydimethylsiloxane matrix for the first time to prepare phase change composites at different loadings. Furthermore, the composites containing the nanocapsules were systematically compared with the composites containing the paraffin@SiO2 microcapsules synthesized using an acidic catalyst. It is shown that, at every identical mass fraction, the composites containing the nanocapsules not only possessed larger latent heat than those containing the microcapsules, but also exhibited higher thermal conductivity and lower hardness. The enhancement in thermal conductivity as well as the decline in hardness for the composite containing the nanocapsules are revealed to originate from a larger amount of hydroxyl groups at the surfaces of the nanocapsules than the microcapsules, which could form more hydrogen bonds with the polymer matrix. This bonding favored the interfacial heat conductance between the nanocapsules and the matrix together with decreasing the crosslinking density of the matrix. Subsequently, composites with enhanced thermal conductivity were developed by combining the nanocapsules with a BN filler. By evaluating the performance for chip heat dissipation, it was found that, when the chip was heated at a power of 10 W, the incorporation of the paraffin@SiO2 nanocapsules at a loading of 36 wt% into the polymer matrix made a remarkable decrease in the chip equilibrium temperature by 31.7 °C, and a further decline by 8.9 °C occurred when combined with 16 wt% BN. This work sheds light on facilitating the interfacial heat conductance between phase change capsules and the polymer matrix by hydrogen bonding.

Clinical Characteristics of Posner-Schlossman Syndrome Patients in China.

Background: To explore the clinical characteristics of Posner-Schlossman syndrome (PSS) patients in China, especially the risk factors associated with the dynamic changes of corneal endothelial cell (CEC) density and retinal nerve fiber layer (RNFL) thickness during the course of the disease. Methods: In this retrospective cohort study, patients consequently suffering from PSS were recruited. Patients data including gender, age of onset, frequency of attack, and disease duration. We performed detailed ophthalmology examinations including intraocular pressure (IOP), best-corrected visual acuity (BCVA), slit-lamp examination, optical coherence tomography (OCT), assessing RNFL thickness, and determined CEC density in the outpatient department. The unaffected eyes served as control. Results: We recruited 121 patients (eyes, n = 125), including 69 (57.0%) males and 52 (43.0%) females with Posner-Schlossman syndrome. The age of first-onset was 33.81 ± 13.63 years old. The majority of these patients were aged 20-50 years (80.2%). The peak IOP was 47.67 ± 13.31 mmHg in the affected eyes. The frequency of PSS attack was 2.7 ± 3.7 times per year. The disease duration was 9.5 ± 10.4 years. Among all patients, there were 27 (22.31%) patients with a history of digestive disorders, 25 (20.66%) patients with a history of allergies, and 24 (19.83%) patients with a history of infectious disease. In the affected eyes, CEC density (2532.94 ± 490.83/mm2 vs. 2777.13 ± 356.87/mm2, p < 0.001), the RNFL thickness of four quadrants (superior 105.46 ± 29.86 µm vs. 121.33 ± 17.30 µm, p < 0.001; temporal 74.58 ± 22.21 µm vs. 81.94 ± 18.20 µm, p = 0.009; inferior 110.50 ± 33.42 µm vs. 128.29 ± 14.39 µm, p < 0.001; nasal 54.85 ± 14.48 µm vs. 63.49 ± 15.40 µm, p < 0.001) and central papillary (87.11 ± 21.18 µm vs. 99.18 ± 7.97 µm, p < 0.001) were significantly reduced compared to the fellow eyes. The disease duration and recurrent frequency were significantly associated with CEC density (p < 0.001 and p = 0.029) and the disease duration was significantly associated with RNFL thickness (p = 0.002). Conclusions: In this study, CEC loss and RNFL thinning were present in the affected eyes. Our results indicated that disease duration and recurrent frequency played an important role in the progression of PSS.

Experimental Study and Calculation Methods of Shear Capacity for High-Strength Reinforced Concrete Full-Scale Deep Beams.

The shear behavior of 8 high-strength concrete full-scale deep beams with high-strength steel bars was studied. The depth beam size was 100 mm × 900 mm × 2200 mm, the test parameters included the shear span-to-depth ratio (λ = 0.9, 0.6, 0.3), longitudinal reinforcement ratio (ρs=0.66%, 1.06%, 1.26%) and stirrup reinforcement ratio (ρsv = 0, 0.26%, 0.34%, 0.5%). The ratio of the cracking load of the inclined section to the ultimate load is between 30% and 50%, and the bending deformation of the deep beam is small, showing the characteristics of brittle failure for deep beams. Under the action of a concentrated load, the failure mode of deep beams with a small shear span ratio is the failure of the diagonal compression struts, which is very different from that of shallow beams with a large shear span ratio. With the increase of shear span ratio from 0.3 to 0.9, the ultimate shear capacity of deep beams decreases by 19.33%. With the increase of longitudinal reinforcement ratio from 0.67% to 1.27%, the ultimate shear capacity of deep beams increased by 45.02%. With the increase of vertical stirrup reinforcement ratio from 0% to 0.5%, the ultimate shear capacity of deep beams increased by 8.93%. Increasing the area of longitudinal bars or stirrups limited the transverse tensile strain of the compression struts, which is conducive to improving the compressive strength of the compression struts of deep beams and then improving the bearing capacity of deep beams. The strut-and-tie model (STM) is more suitable for analyzing the shear capacity of deep beams. The calculation methods for calculating the shear capacity of deep beams were compared with ACI 318-19, CSA A23 3-19, EN 1992-1-1:2004, Tan-Tan model, Tan-Cheng model, softened STM (SSTM) and simplified SSTM (SSSTM). The results showed that the shear capacity of deep beams could be well predicted by reasonably determining the STM parameters.

Effects of Extracellular Vesicles from Osteogenic Differentiated Human BMSCs on Osteogenic and Adipogenic Differentiation Capacity of Naïve Human BMSCs.

Osteoporosis, or steroid-induced osteonecrosis of the hip, is accompanied by increased bone marrow adipogenesis. Such a disorder of adipogenic/osteogenic differentiation, affecting bone-marrow-derived mesenchymal stem cells (BMSCs), contributes to bone loss during aging. Here, we investigated the effects of extracellular vesicles (EVs) isolated from human (h)BMSCs during different stages of osteogenic differentiation on the osteogenic and adipogenic differentiation capacity of naïve (undifferentiated) hBMSCs. We observed that all EV groups increased viability and proliferation capacity and suppressed the apoptosis of naïve hBMSCs. In particular, EVs derived from hBMSCs at late-stage osteogenic differentiation promoted the osteogenic potential of naïve hBMSCs more effectively than EVs derived from naïve hBMSCs (naïve EVs), as indicated by the increased gene expression of COL1A1 and OPN. In contrast, the adipogenic differentiation capacity of naïve hBMSCs was inhibited by treatment with EVs from osteogenic differentiated hBMSCs. Proteomic analysis revealed that osteogenic EVs and naïve EVs contained distinct protein profiles, with pro-osteogenic and anti-adipogenic proteins encapsulated in osteogenic EVs. We speculate that osteogenic EVs could serve as an intercellular communication system between bone- and bone-marrow adipose tissue, for transporting osteogenic factors and thus favoring pro-osteogenic processes. Our data may support the theory of an endocrine circuit with the skeleton functioning as a ductless gland.

COL3A1 and MMP9 Serve as Potential Diagnostic Biomarkers of Osteoarthritis and Are Associated With Immune Cell Infiltration.

BACKGROUND: Osteoarthritis (OA) is one of the most common age-related degenerative diseases. In recent years, some studies have shown that pathological changes in the synovial membrane occur earlier than those in the cartilage in OA. However, the molecular mechanism of synovitis in the pathological process of OA has not been elucidated. This study aimed to identify novel biomarkers associated with OA and to emphasize the role of immune cells in the pathogenesis of OA. METHODS: Microarray datasets were obtained from the Gene Expression Omnibus (GEO) and ArrayExpress databases and were then analyzed using R software. To determine differential immune cell subtype infiltration, the CIBERSORT deconvolution algorithm was used. Quantitative reverse transcription PCR (qRT-PCR) was used to determine the relative expressions of selected genes. Besides, Western blotting was used to assess the protein expression levels in osteoarthritic chondrocytes. RESULTS: After analyzing the database profiles, two potential biomarkers, collagen type 3 alpha 1 chain (COL3A1), and matrix metalloproteinase 9 (MMP9), associated with OA were discovered, which were confirmed by qRT-PCR and Western blotting. Specifically, the results revealed that, as the concentration of IL-1ß increased, so did the gene and protein expression levels of COL3A1 and MMP9. CONCLUSION: The findings provide valuable information and direction for future research into novel targets for OA immunotherapy and diagnosis and aids in the discovery of the underlying biological mechanisms of OA pathogenesis.

Curcumin-primed human BMSC-derived extracellular vesicles reverse IL-1ß-induced catabolic responses of OA chondrocytes by upregulating miR-126-3p.

BACKGROUND: Curcumin has anti-inflammatory effects and qualifies as a potential candidate for the treatment of osteoarthritis (OA). However, curcumin has limited bioavailability. Extracellular vesicles (EVs) are released by multiple cell types and act as molecule carrier during intercellular communication. We assume that EVs can maintain bioavailability and stability of curcumin after encapsulation. Here, we evaluated modulatory effects of curcumin-primed human (h)BMSC-derived EVs (Cur-EVs) on IL-1ß stimulated human osteoarthritic chondrocytes (OA-CH). METHODS: CellTiter-Blue Viability- (CTB), Caspase 3/7-, and live/dead assays were used to determine range of cytotoxic curcumin concentrations for hBMSC and OA-CH. Cur-EVs and control EVs were harvested from cell culture supernatants of hBMSC by ultracentrifugation. Western blotting (WB), transmission electron microscopy, and nanoparticle tracking analysis were performed to characterize the EVs. The intracellular incorporation of EVs derived from PHK26 labeled and curcumin-primed or control hBMSC was tested by adding the labeled EVs to OA-CH cultures. OA-CH were pre-stimulated with IL-1ß, followed by Cur-EV and control EV treatment for 24 h and subsequent analysis of viability, apoptosis, and migration (scratch assay). Relative expression of selected anabolic and catabolic genes was assessed with qRT-PCR. Furthermore, WB was performed to evaluate phosphorylation of Erk1/2, PI3K/Akt, and p38MAPK in OA-CH. The effect of hsa-miR-126-3p expression on IL-1ß-induced OA-CH was determined using CTB-, Caspase 3/7-, live/dead assays, and WB. RESULTS: Cur-EVs promoted viability and reduced apoptosis of IL-1ß-stimulated OA-CH and attenuated IL-1ß-induced inhibition of migration. Furthermore, Cur-EVs increased gene expression of BCL2, ACAN, SOX9, and COL2A1 and decreased gene expression of IL1B, IL6, MMP13, and COL10A1 in IL-1ß-stimulated OA-CH. In addition, phosphorylation of Erk1/2, PI3K/Akt, and p38 MAPK, induced by IL-1ß, is prevented by Cur-EVs. Cur-EVs increased IL-1ß-reduced expression of hsa-miR-126-3p and hsa-miR-126-3p mimic reversed the effects of IL-1ß. CONCLUSION: Cur-EVs alleviated IL-1ß-induced catabolic effects on OA-CH by promoting viability and migration, reducing apoptosis and phosphorylation of Erk1/2, PI3K/Akt, and p38 MAPK thereby modulating pro-inflammatory signaling pathways. Treatment of OA-CH with Cur-EVs is followed by upregulation of expression of hsa-miR-126-3p which is involved in modulation of anabolic response of OA-CH. EVs may be considered as promising drug delivery vehicles of curcumin helping to alleviate OA.

A pilot study of the precision of digital intraocular pressure measurement during vitrectomy.

AIM: To evaluate the precision of digital intraocular pressure (IOP) measurement in silicone oil (SO) filled eyes during vitrectomy. METHODS: This is a retrospective, single-blind study. Patients who were diagnosed with retinal detachment and scheduled for vitrectomy with SO injection were consecutively enrolled. During the vitrectomy, IOP was digitally measured and then by a rebound tonometer (IcarePRO). The rebound tonometer readings were masked to the surgeons. The digitally measured IOP and that of rebound tonometer were compared, and the inter-methods agreement was assessed. The absolute deviation in IOP values between these two methods (ΔIOP) was also calculated, and correlations between ΔIOP and refractive status, lens status and levels of surgeons' experience were analyzed. RESULTS: A total of 131 patients (131 eyes) were recruited, with a mean age of 51.0±16.1y. There was no significant difference in IOPs between digital measurement and the rebound tonometer (15.6±4.3 vs 15.7±5.1 mm Hg; t=0.406, P=0.686). Intraclass correlation coefficients (ICC) analysis indicated a strong correlation between these two measurements (ICC=0.830, P<0.001). The mean ΔIOP was 2.0±1.9 mm Hg (range: 0-12.8 mm Hg), with 98 eyes (74.8%) had the ΔIOP within 3 mm Hg. ΔIOP was found to be negatively correlated with levels of surgeons' experience (r=-0.183; P=0.037), but not with the refractive status or lens status of the patients (both P>0.05). CONCLUSION: For experienced surgeons, the digital IOP measurement may be an acceptable technique for IOP measurement in SO filled eyes during vitrectomy. However, its use by inexperienced surgeons should be taken with caution.

SOX9 Knockout Induces Polyploidy and Changes Sensitivity to Tumor Treatment Strategies in a Chondrosarcoma Cell Line.

As most chemotherapeutic drugs are ineffective in the treatment of chondrosarcoma, we studied the expression pattern and function of SOX9, the master transcription factor for chondrogenesis, in chondrosarcoma, to understand the basic molecular principles needed for engineering new targeted therapies. Our study shows an increase in SOX9 expression in chondrosarcoma compared to normal cartilage, but a decrease when the tumors are finally defined as dedifferentiated chondrosarcoma (DDCS). In DDCS, SOX9 is almost completely absent in the non-chondroid, dedifferentiated compartments. CRISPR/Cas9-mediated knockout of SOX9 in a human chondrosarcoma cell line (HTB94) results in reduced proliferation, clonogenicity and migration, accompanied by an inability to activate MMP13. In contrast, adhesion, apoptosis and polyploidy formation are favored after SOX9 deletion, probably involving BCL2 and survivin. The siRNA-mediated SOX9 knockdown partially confirmed these results, suggesting the need for a certain SOX9 threshold for particular cancer-related events. To increase the efficacy of chondrosarcoma therapies, potential therapeutic approaches were analyzed in SOX9 knockout cells. Here, we found an increased impact of doxorubicin, but a reduced sensitivity for oncolytic virus treatment. Our observations present novel insight into the role of SOX9 in chondrosarcoma biology and could thereby help to overcome the obstacle of drug resistance and limited therapy options.

Experimental Study on Shear Capacity of High Strength Reinforcement Concrete Deep Beams with Small Shear Span-Depth Ratio.

This study aimed to investigate the shear capacity performance for eight deep beams with HTRB600 reinforced high strength concrete under concentrated load to enable a better understanding of the effects of shear span-depth ratio, longitudinal reinforcement ratio, vertical stirrup ratio and in order to improve design procedures. The dimension of eight test specimens is 1600 mm × 200 mm × 600 mm. The effective span to height ratio l0/h is 2.0, the shear span-depth ratio λ is 0.3, 0.6 and 0.9, respectively. In addition, the longitudinal reinforcement ratio ρs is set to 0.67%, 1.05%, 1.27%, and the vertical stirrup ratio is taken to be 0%, 0.25%, 0.33%, 0.5%. Through measuring the strain of steel bar, the strain of concrete and the deflection of mid-span, the characteristics of the full process of shear capacity, the failure mode and the load deflection deformation curve were examined. The test results showed that the failure mode of deep beams with small shear span-depth ratio is diagonal compression failure, which is influenced by the layout and quantity of web reinforcement. The diagonal compression failure could be classified into two forms: crushing-strut and diagonal splitting. With decreasing of shear span-depth ratio and increasing longitudinal reinforcement ratio, the shear capacity of deep beams increases obviously, while the influence of vertical web reinforcement ratio on shear capacity is negligible. Finally, the shear capacity of eight deep beams based on GB 50010-2010 is calculated and compared with the calculation results of ACI 318-14, EN 1992-1-1:2004 and CSA A23.3-04, which are based on strut-and-tie model. The obtained results in this paper show a very good agreement with GB50010-2010 and ACI 318-14, while the results of EN 1992-1-1:2004 and CSA A23.3-04 are approved to be conservative.

hBMSC-Derived Extracellular Vesicles Attenuate IL-1ß-Induced Catabolic Effects on OA-Chondrocytes by Regulating Pro-inflammatory Signaling Pathways.

Background: Human bone marrow-derived mesenchymal stromal cells (hBMSCs) provide a promising therapeutic approach in the cell-based therapy of osteoarthritis (OA). However, several disadvantages evolved recently, including immune responses of the host and regulatory hurdles, making it necessary to search for alternative treatment options. Extracellular vesicles (EVs) are released by multiple cell types and tissues into the extracellular microenvironment, acting as message carriers during intercellular communication. Here, we investigate putative protective effects of hBMSC-derived EVs as a cell-free approach, on IL-1ß-stimulated chondrocytes obtained from OA-patients. Methods: EVs were harvested from the cell culture supernatant of hBMSCs by a sequential ultracentrifugation process. Western blot, scanning electron microscopy (SEM), and nanoparticle tracking analysis (NTA) were performed to characterize the purified particles as EVs. Intracellular incorporation of EVs, derived from PHK26-labeled hBMSCs, was tested by adding the labeled EVs to human OA chondrocytes (OA-CH), followed by fluorescence microscopy. Chondrocytes were pre-stimulated with IL-1ß for 24 h, followed by EVs treatment for 24 h. Subsequently, proliferation, apoptosis, and migration (wound healing) were analyzed via BrdU assay, caspase 3/7 assay, and scratch assay, respectively. With qRT-PCR, the relative expression level of anabolic and catabolic genes was determined. Furthermore, immunofluorescence microscopy and western blot were performed to evaluate the protein expression and phosphorylation levels of Erk1/2, PI3K/Akt, p38, TAK1, and NF-κB as components of pro-inflammatory signaling pathways in OA-CH. Results: EVs from hBMSCs (hBMSC-EVs) promote proliferation and reduce apoptosis of OA-CH and IL-1ß-stimulated OA-CH. Moreover, hBMSC-EVs attenuate IL-1ß-induced reduction of chondrocyte migration. Furthermore, hBMSC-EVs increase gene expression of PRG4, BCL2, and ACAN (aggrecan) and decrease gene expression of MMP13, ALPL, and IL1ß in OA-CH. Notably, COL2A1, SOX9, BCL2, ACAN, and COMP gene expression levels were significantly increased in IL-1ß+ EV groups compared with those IL-1ß groups without EVs, whereas the gene expression levels of COLX, IL1B, MMP13, and ALPL were significantly decreased in IL-1ß+ EV groups compared to IL-1ß groups without EVs. In addition, the phosphorylation status of Erk1/2, PI3K/Akt, p38, TAK1, and NF-κB signaling molecules, induced by IL-1ß, is prevented by hBMSC- EVs. Conclusion: EVs derived from hBMSCs alleviated IL-1ß-induced catabolic effects on OA-CH via promoting proliferation and migration and reducing apoptosis, probably via downregulation of IL-1ß-activated pro-inflammatory Erk1/2, PI3K/Akt, p38, TAK1, and NF-κB signaling pathways. EVs released from BMSCs may be considered as promising cell-free intervention strategy in cartilage regenerative medicine, avoiding several adverse effects of cell-based regenerative approaches.

Influence of Extracellular Vesicles Isolated From Osteoblasts of Patients With Cox-Arthrosis and/or Osteoporosis on Metabolism and Osteogenic Differentiation of BMSCs.

Background: Studies with extracellular vesicles (EVs), including exosomes, isolated from mesenchymal stem cells (MSC) indicate benefits for the treatment of musculoskeletal pathologies as osteoarthritis (OA) and osteoporosis (OP). However, little is known about intercellular effects of EVs derived from pathologically altered cells that might influence the outcome by counteracting effects from "healthy" MSC derived EVs. We hypothesize, that EVs isolated from osteoblasts of patients with hip OA (coxarthrosis/CA), osteoporosis (OP), or a combination of both (CA/OP) might negatively affect metabolism and osteogenic differentiation of bone-marrow derived (B)MSCs. Methods: Osteoblasts, isolated from bone explants of CA, OP, and CA/OP patients, were compared regarding growth, viability, and osteogenic differentiation capacity. Structural features of bone explants were analyzed via µCT. EVs were isolated from supernatant of naïve BMSCs and CA, OP, and CA/OP osteoblasts (osteogenic culture for 35 days). BMSC cultures were stimulated with EVs and subsequently, cell metabolism, osteogenic marker gene expression, and osteogenic differentiation were analyzed. Results: Trabecular bone structure was different between the three groups with lowest number and highest separation in the CA/OP group. Viability and Alizarin red staining increased over culture time in CA/OP osteoblasts whereas growth of osteoblasts was comparable. Alizarin red staining was by trend higher in CA compared to OP osteoblasts after 35 days and ALP activity was higher after 28 and 35 days. Stimulation of BMSC cultures with CA, OP, and CA/OP EVs did not affect proliferation but increased caspase 3/7-activity compared to unstimulated BMSCs. BMSC viability was reduced after stimulation with CA and CA/OP EVs compared to unstimulated BMSCs or stimulation with OP EVs. ALP gene expression and activity were reduced in BMSCs after stimulation with CA, OP, and CA/OP EVs. Stimulation of BMSCs with CA EVs reduced Alizarin Red staining by trend. Conclusion: Stimulation of BMSCs with EVs isolated from CA, OP, and CA/OP osteoblasts had mostly catabolic effects on cell metabolism and osteogenic differentiation irrespective of donor pathology and reflect the impact of tissue microenvironment on cell metabolism. These catabolic effects are important for understanding differences in effects of EVs on target tissues/cells when harnessing them as therapeutic drugs.

Extracellular Vesicles in Musculoskeletal Pathologies and Regeneration.

The incidence of musculoskeletal diseases is steadily increasing with aging of the population. In the past years, extracellular vesicles (EVs) have gained attention in musculoskeletal research. EVs have been associated with various musculoskeletal pathologies as well as suggested as treatment option. EVs play a pivotal role in communication between cells and their environment. Thereby, the EV cargo is highly dependent on their cellular origin. In this review, we summarize putative mechanisms by which EVs can contribute to musculoskeletal tissue homeostasis, regeneration and disease, in particular matrix remodeling and mineralization, pro-angiogenic effects and immunomodulatory activities. Mesenchymal stromal cells (MSCs) present the most frequently used cell source for EV generation for musculoskeletal applications, and herein we discuss how the MSC phenotype can influence the cargo and thus the regenerative potential of EVs. Induced pluripotent stem cell-derived mesenchymal progenitor cells (iMPs) may overcome current limitations of MSCs, and iMP-derived EVs are discussed as an alternative strategy. In the last part of the article, we focus on therapeutic applications of EVs and discuss both practical considerations for EV production and the current state of EV-based therapies.

Role of microRNA-145 in protection against myocardial ischemia/reperfusion injury in mice by regulating expression of GZMK with the treatment of sevoflurane.

This study aims to investigate the role of microRNA-145 (miR-145) in protection against myocardial ischemia/reperfusion (I/R) injury in mice by regulating expression of granzyme K (GZMK) with the treatment of sevoflurane. The mice model of myocardial I/R injury was established by left coronary artery ligation. The expression of miR-145 and GZMK in myocardial tissues of mice was detected by Reverse transcription quantitative polymerase chain reaction and western blot analysis. The changes of the cardiac function and hemodynamics, pathological changes of myocardial tissues, the ultrastructure of cardiomyocytes, myocardial infarction area, and cardiomyocyte apoptosis were observed. The expression of the apoptosis-related protein cleaved-caspase-3, Bax, and Bcl-2 was detected by western blot analysis. The levels of malondialdehyde, myeloperoxidase, superoxide dismutase in myocardial tissues were detected by spectrophotometric colorimetry. The levels of interleukin-1ß (IL-1ß), IL-6, and tumor necrosis factor-α in the serum of mice were detected by the enzyme-linked immunosorbent assay. The level of oxidative stress and the expression of inflammatory factors increased in mice with myocardial I/R injury. Sevoflurane postconditioning could reduce myocardial I/R injury in mice. Sevoflurane postconditioning may protect myocardial I/R injury through miR-145-regulation of GZMK in mice. Inhibition of miR-145 expression could reduce the protective effect of sevoflurane posttreatment on myocardial I/R injury in mice. Low expression of GZMK could attenuate the inhibitory effect of miR-145 on myocardial I/R injury after sevoflurane treatment in mice. Our study suggests that sevoflurane postconditioning may protect against myocardial I/R injury by upregulating miR-145 expression and downregulating GZMK expression.

The clinical feature of myocilin Y437H mutation in a Chinese family with primary open-angle glaucoma.

PURPOSE: To characterise the genotype(s), phenotype(s) and age-related penetrance of glaucoma in a Chinese family with primary open-angle glaucoma (POAG). METHODS: Recruited from a Chinese family spanning four generations, 7 individuals with POAG, 1 with ocular hypertension (OHT) and 14 unaffected individuals were included in this study. Genotypic investigation included sequencing of mutation sites using a glaucoma panel in combination with high-throughput sequencing and validated using Sanger sequencing. Phenotypic characterisation included investigation into patient medical history and physical examination. RESULTS: Eight (36.4%) family members harboured heterozygous Y437H mutation, of whom seven (87.5%) were glaucomatous and one (12.5%) had OHT. The mean age of POAG diagnosis was 30.85±7.13 years. The mean highest recorded intraocular pressure (IOP) was 46.57±6.53 mm Hg. They all had complained of symptoms associated with vision and pain. Four (57.1%) patients presented blindness. Five eyes (62.5%) presented with severe and three eyes with moderate visual field defects. Most of them underwent surgery on average 1.29±2.36 years after diagnosis, and the mean IOP at study was 17.95±7.23 mm Hg, with an average of 0.92±0.86 eye-drops. The patient with OHT was treated with latanoprost only and her IOP was well controlled. Age-related glaucoma penetrance was 0% in individuals under the age of 20 years, 50% at ages 20-35 years, 75% at ages 31-35 years and 87.5% over 45 years. CONCLUSION: A novel MYOC mutation (c.1309T>C, p.Y437H) in a Chinese family with POAG was identified which was associated with a phenotype characterised by severe visual impairment, frequent surgical intervention requirement and relatively high penetrance.

[Dynesys and posterior decompression and fusion internal fixation for the treatment of lumbar degenerative diseases: a systematic review].

OBJECTIVE: Meta analysis was used to evaluate the efficacy and safety of Dynesys and posterior decompression and fusion internal fixation for lumbar degenerative diseases. METHODS: The computer was used to retrieve the Cochrane library, Medline, Embase, CNKI, Wanfang database and Chinese biomedical literature database; and the references and main Chinese and English Department of orthopedics journals were manually searched. All the prospective or retrospective comparative studies on the clinical efficacy and safety of Dynesys and posterior decompression and fusion internal fixation were collected, so as to evaluate the methodological quality of the study and to extract the data. The RevMan 5.2 software provided by Cochrane collaboration was used for systematic evaluation. RESULTS: A total of 9 clinical studies were included, including 3 prospective randomized controlled trials(RCT) and 6 retrospective controlled observational studies, which included 692 patients, with 336 cases in Dynesys group, and 356 cases in posterior decompression and fusion internal fixation (PLIF) group. The results showed that compared with PLIF, Dynesys system significantly decreased operation time(P<0.01), intraoperative blood loss (P<0.01). Both Dynesys and PLIF groups experienced improved ODI and back/leg pain VAS scores at final follow-up, and no statistically significant difference was noted according to the two surgical procedures(P>0.05). Dynesys could remain the range of motion (ROM) of surgical segments with less increased ROM of adjacent segments compared with that of PLIF group(P<0.01). Regarding the disc height of surgical segments, no statistically significant difference was noted according to the two groups(P>0.05), but postoperative complications incidence rate in PLIF group was higher than that in Dynesys group (P<0.05). CONCLUSIONS: Both Dynesys system and PLIF can improve clinical outcomes of lumbar degenerative diseases effectively. Compared with PLIF, Dynesys could remain the range of motion(ROM) of surgical segments with less increased ROM of adjacent segments and lower complication incidence rate. But the ability to prevent adjacent segments degeneration needs more RCTs with long-term follow-up to confirm.

Changes of soil bacterial activities and functions after different N additions in a temperate forest.

It has been shown that different nitrogen (N) addition led to various influences on soil microbial activities in forest ecosystems; however, the changes of bacteria were still unclear. In this work, inorganic N (NH4NO3) and organic N (urea and glycine) were fertilized with different ratios (5:0, 1:4, 3:2, 2:3, and 1:4) on temperate forest soils, while fungicide (cycloheximide) was simultaneously added on half of each treatment to inhibit fungal activities (leaving only bacteria). After a 3-year field experiment, soil samples were harvested, then microbial enzymatic activities involved in carbon (C), and N and phosphorus (P) cycles were determined. Under laboratory conditions, four purified bacteria which were isolated from sample site had been inoculated in sterilized soils under different N types and enzymatic activities were assayed after 90-day incubation. The results showed that cellulase and polyphenol oxidase activities of non-fungicide-added treatments increased after N addition and greater organic N accelerated the increases. However, these enzymatic activities of fungicide-added treatments were not significantly influenced by N addition and N types. It may be due to the insufficient ability of bacteria to synthesize enough enzymes to decompose complex organic C (such as cellulose and lignin) into available compound, although N-limitation was alleviated. Alkaline phosphatase activities increased after N addition in both non-fungicide-added and fungicide-added treatments, and the acceleration on bacterial alkaline phosphatase activities was even greater. Furthermore, organic N showed at least 2.5 times promotion on bacteria alkaline phosphatase than those of inorganic N, which indicated greater alleviation of bacterial P-limitation after the addition of organic N. All the results indicated that soil bacteria may be seriously limited by soil available C but become the dominant decomposer of the complex P compounds after N addition, particularly greater organic N.