Search

Main Conclusion: This study reveals miRNA indirect regulation of C4 genes in sugarcane through transcription factors, highlighting potential key regulators like SsHAM3a. C4 photosynthesis is crucial for the high productivity and biomass of sugarcane, however, the miRNA regulation of C4 genes in sugarcane remains elusive. We have identified 384 miRNAs along the leaf gradients, including 293 known miRNAs and 91 novel miRNAs. Among these, 86 unique miRNAs exhibited differential expression patterns, and we identified 3511 potential expressed targets of these differentially expressed miRNAs (DEmiRNAs). Analyses using Pearson correlation coefficient (PCC) and Gene Ontology (GO) enrichment revealed that targets of miRNAs with positive correlations are integral to chlorophyll-related photosynthetic processes. In contrast, negatively correlated pairs are primarily associated with metabolic functions. It is worth noting that no C4 genes were predicted as targets of DEmiRNAs. Our application of weighted gene co-expression network analysis (WGCNA) led to a gene regulatory network (GRN) suggesting miRNAs might indirectly regulate C4 genes via transcription factors (TFs). The GRAS TF SsHAM3a emerged as a potential regulator of C4 genes, targeted by miR171y and miR171am, and exhibiting a negative correlation with miRNA expression along the leaf gradient. This study sheds light on the complex involvement of miRNAs in regulating C4 genes, offering a foundation for future research into enhancing sugarcane's photosynthetic efficiency. [ABSTRACT FROM AUTHOR]

Highlights: A multifactorial and oriented scaffolds was constructed to stimulate osteochondral regeneration. This is the first demonstration that both drug and gene nanoparticles were spatially deposited on a patterned film through metal-ligand interactions. For the first time, film-rolling approach was employed to construct osteochondral plug to mimick the Haversian canal structure of natural bone. The integrated repair of bone and cartilage boasts advantages for osteochondral restoration such as a long-term repair effect and less deterioration compared to repairing cartilage alone. Constructing multifactorial, spatially oriented scaffolds to stimulate osteochondral regeneration, has immense significance. Herein, targeted drugs, namely kartogenin@polydopamine (KGN@PDA) nanoparticles for cartilage repair and miRNA@calcium phosphate (miRNA@CaP) NPs for bone regeneration, were in situ deposited on a patterned supramolecular-assembled 2-ureido-4 [lH]-pyrimidinone (UPy) modified gelation hydrogel film, facilitated by the dynamic and responsive coordination and complexation of metal ions and their ligands. This hydrogel film can be rolled into a cylindrical plug, mimicking the Haversian canal structure of natural bone. The resultant hydrogel demonstrates stable mechanical properties, a self-healing ability, a high capability for reactive oxygen species capture, and controlled release of KGN and miR-26a. In vitro, KGN@PDA and miRNA@CaP promote chondrogenic and osteogenic differentiation of mesenchymal stem cells via the JNK/RUNX1 and GSK-3β/β-catenin pathways, respectively. In vivo, the osteochondral plug exhibits optimal subchondral bone and cartilage regeneration, evidenced by a significant increase in glycosaminoglycan and collagen accumulation in specific zones, along with the successful integration of neocartilage with subchondral bone. This biomaterial delivery approach represents a significant toward improved osteochondral repair. [ABSTRACT FROM AUTHOR]

Objective: This study is aimed to investigate the clinical outcomes of a novel SSPS for fixation of the comminuted coronoid fracture. Methods: A retrospective study was carried out in the patients with comminuted fractures of the coronoid treated by SPSS fixation between January 2014 and December 2018. A total of 17 patients (17 sides) was included in our study, including 11 male and six female, with a mean age range from 18 to 60. All cases started to functional rehabilitation immediately after the operation. Clinical outcomes were evaluated both radiographically and functionally at the follow‐up visit, including the elbow instability, range of motion and Mayo elbow performance score (MEPS). Results: According to the O'Driscoll classification system, there was two side of type 1.2, two of type 2.1, four of type 2.2, three of type 2.3, two of 3.1 and four of type 3.2. The surgery was carried out by Kocher and anteromedial approach in 12 patients, posterior and anteromedial approach in four, anterior approach in one. The average operation time and intraoperative blood loss was 129.41±43.87 min and 115.29±104.65 ml. The median follow‐up time was 9 months (range, 6 to 15 months). The mean flexion, extension, pronation and supination motion was 138.76±8.67 degrees, 20.00±13.58, 82.94±5.32and 74.12±14.39 respectively at final follow up. The mean MEPS score was 89.76±8.46, including 11 excellent, 3 good and 3 fair result. The mean VAS score was 1.94±0.97. The mean union time of coronoid fractures was 2.77±0.31 months according to the established standard of healing. There were no significant differences in clinical outcomes among groups according to the O'Driscoll classification (P >.05) and ligament repair strategy (P >.05). No patient underwent instability or dislocation of the elbow during follow up. There were two cases with mild ulnar nerve symptoms which recovered totally at follow up. Meanwhile, there were three cases with heterotopic ossification of the elbow. Conclusion: Our findings demonstrated that the SSPS can provide a reliable fixation for the comminuted coronoid fracture with satisfactory clinical outcomes. [ABSTRACT FROM AUTHOR]

Objective: To tackle the challengeable dilemma of delayed femoral fracture, a technique of fixator‐assisted closed reduction and intramedullary nailing at one stage was introduced herein and its clinical results were investigated. Methods: A retrospective study was conducted on delayed femoral shaft fracture between February 2008 and January 2017. The multiple injured patients aged from 18 to 60 years with delayed femoral fracture was included. All patients were treated by one‐stage internal fixation technique and followed up for more than 1 year. Outcome measures including the operation time, intraoperative blood loss and limb alignment, healing time of fracture, visual analog scale (VAS), and range of knee motion were recorded and evaluated. Results: A total of 13 patients (16 sides) with a mean injury severity score (ISS) of 32.77 ± 9.98 (range, 19 to 52) participated in the investigation. The median length of time‐after‐fracture was 38 days (range, 21 to 110 days). The average shortening distance of the fracture ends was 35.48 ± 19.24 mm (range, 10.00 to 79.00 mm). The average surgery time was 192.19 ± 29.38 min for unilateral femoral fracture, with blood loss of 587.50 ± 232.02 ml. The postoperative discrepancy of lower limb was 3.87 ± 2.52 mm. No patient had vascular and neurologic complications due to the lengthening. All fractures healed successfully with a mean time of 2.98 ± 0.57 months. The mean VAS and maximal knee flexion were 1.63 ± 1.09 and 131.25° ± 5.32° at final follow‐up, respectively. Conclusions: Our findings indicated that fixator‐assisted closed reduction and intramedullary nailing at one stage is an effective treatment for delayed femoral fracture with satisfactory functional recovery. [ABSTRACT FROM AUTHOR]

Intervertebral disc (IVD) degenerative disease is a common health problem worldwide. Administration of mesenchymal stem cells (MSCs) in intervertebral disc degeneration (IVDD) has been widely explored in recent years. However, transplantation of MSCs is restricted by several factors. Currently, paracrine signaling is one of the main mechanisms by which MSCs play a therapeutic role in disc regeneration. Extracellular vehicles (EVs) are the main paracrine products of MSCs. They show great potential as an effective alternative to MSCs and play immunomodulation roles such as anti-inflammatory effects, antioxidative stress, antiapoptosis, and antiextracellular matrix (ECM) degradation during treatment of IVDD. This review focuses on the immunomodulatory effect of MSC EVs and their potential applications. [ABSTRACT FROM AUTHOR]

Gold nanorods (GNRs) are intensively explored for the application in cancer therapy, which has motivated the development of photothermal therapy (PTT) multifunctional nanoplatforms based on GNRs to cure osteosarcoma (OS). However, the major limitations include the toxicity of surface protectants of GNRs, unsatisfactory targeting therapy, and the resistant effects of photothermal-induced autophagy, so the risk of relapse and metastasis of OS increase. In the present study, the GNR multifunctional nanoplatforms were designed and synthesized to deliver transcription factor EB (TFEB)-siRNA–targeting autophagy; then, the resistance of autophagy to PTT and the pH-sensitive cell-penetrating membrane peptide (CPP) was weakened, which could improve the tumor-targeting ability of the GNR nanoplatforms and realize an efficient synergistic effect for tumor treatment. Meanwhile, it is worth noting that the GNR nanoplatform groups have anti-lung metastasis of OS. This study provides a new reference to improve the efficacy of OS clinically. [ABSTRACT FROM AUTHOR]

Articular cartilage regeneration remains a major challenge in orthopedics. Noncanonical Wnt5a is a particularly attractive growth factor in this context; Wnt5a inhibits chondrocyte hypertrophy but maintains chondrogenesis. We designed a novel, vertically oriented‐collagen scaffold. The effect of Wnt5a on MSCs and chondrocytes and the therapeutic effects of the Wnt5a/oriented‐collagen scaffold in terms of osteochondral repair and cartilage integration were evaluated. In vitro, the proliferation, migration, and differentiation of MSCs and chondrocytes treated with Wnt5a, and the mechanisms thereof, were assessed. mRNA microarray analysis was performed to compare the expression profiles of MSCs before and after Wnt5a treatment. In vivo, full‐thickness cylindrical osteochondral defects (4 mm in diameter, 3 mm in depth) were created in the patellar grooves of 24 New Zealand white rabbits and implanted with oriented‐collagen scaffolds (n = 8), Wnt5a/oriented‐collagen scaffolds (n = 8), or nothing (n = 8). After 6 and 12 weeks, integration and tissue responses were evaluated. The proliferation, migration, chondrogenic differentiation, and extracellular matrix formation of/by MSCs and chondrocytes improved greatly after treatment with Wnt5a. Western blotting showed that the PI3K/AKT/JNK signaling pathway was activated. Microarray analysis revealed that the Wnt5a group exhibited a significant upregulation of the PI3K pathway. Reactome GSEA pathway interaction analysis revealed that such upregulation was associated with collagen and extracellular matrix formation. In vivo, the Wnt5a/oriented‐collagen scaffold group exhibited optimal interface integration, cartilage regeneration, and collagenous fiber arrangement, accompanied by significantly increased glycosaminoglycan and collagen accumulations in the zones of regeneration and integration, compared to the other groups. Gene expression analysis showed that the levels of mRNAs encoding genes involved in cartilage formation were significantly increased in the Wnt5a/oriented, collagen scaffold group (all P <.05). Wnt5a promoted the proliferation, migration, and chondrogenic differentiation of MSCs and chondrocytes via the activation of the PI3K/AKT/JNK signaling pathway. The Wnt5a/oriented‐collagen constructs enhanced the structure‐specific regeneration of hyaline cartilage in a rabbit model and may be a promising treatment for the repair of human cartilage defects. [ABSTRACT FROM AUTHOR]

Malignant melanoma has the highest malignancy rate among all skin cancer and is characterized by an insidious onset, high invasion and poor patient prognosis. Yet, the mechanisms involved remain unclear and warrant further investigation. Based on bioinformatic analysis, phospholipase C β2 (PLCB2) has been found to be correlated with melanoma growth. The present study was the first to demonstrate that PLCB2 is a key factor affecting melanoma proliferation and apoptosis. Here, microarray datasets from the publicly available Gene Expression Omnibus (GEO) database were employed, and gene set enrichment analysis (GSEA) was introduced to identify candidate transcription factors. PLCB2 was identified as a crucial gene in the protein-protein interaction (PPI) network. The expression of PLCB2 mRNA in various cancer lines was analyzed by reverse transcription-polymerase chain reaction (RT-PCR). In addition, the proliferation ability and apoptosis rate in human melanoma cells overexpressing or not overexpressing PLCB2 were assessed using colony formation assay, flow cytometry and the Cell Counting Kit-8 (CCK-8) assay. Cell viability and apoptosis-related factors, such as p53, Bcl-2, Bax and caspase-3 were significantly regulated. Knockdown of PLCB2 suppressed the activation of the Ras/Raf/MAPK signaling pathway. In conclusion, knockdown of PLCB2 suppressed cell viability and promoted cell apoptosis by activating the Ras/Raf/MAPK pathway. Thus, PLCB2 may utilized as a potential therapeutic target in patients with melanoma. [ABSTRACT FROM AUTHOR]

Tendon injuries are common and require a long time to heal, and are particularly associated with some adverse problems such as adhesion and rupture. Herein, we aim to develop new bioactive scaffolds endowed with stem cell sheets and growth factors to enable cell migration and proliferation favorable for tendon regeneration in situ. An exogenous basic fibroblast growth factor (bFGF)-loaded fibrin gel was firstly incorporated into the porous network of knitted poly(lactide-co-glycolide) (PLGA) scaffolds and then sheets of mesenchymal stem cells (MSCs) were also integrated into the scaffolds. It was shown that the pores in the knitted PLGA scaffold were readily filled with a complex network of fibrin fiber gel and the fibrin fibers were beneficial for the controlled release of bFGF over a long time period. After transplantation in a critical-size Achilles tendon defect model (7 mm) in the rat right hindlimb, gross observation revealed no immunologic incompatibility or rejection derived from the scaffold systems. It was observed that the MSC sheets contributed directly to tendon regeneration, and exerted an environment-modifying effect on the injuries in situ, consistent with the beneficial effect of bFGF. It was interesting that the knitted PLGA–fibrin gel scaffolds loaded with MSC sheets and bFGF showed the highest expression of tendon-related gene markers and outstanding repair efficacy, including appreciable biomechanical strength and native-like histological microstructures. Therefore, the integration of MSC sheets and bFGF into PLGA/bFGF–fibrin gel scaffolds may stimulate the proliferation and tenogenic differentiation of MSCs in situ and synergistically enhance the injured tendon reconstruction. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Background/Aims: MicroRNAs (miRNAs) are a class of small noncoding RNAs that regulate gene expression by repressing translation or cleaving RNA transcripts in a sequence-specific manner. Downregulated microRNAs and their roles in cancer development have attracted much attention. A growing body of evidence showed that microRNA-133a (miR-133a) has inhibitory effects on cell proliferation, migration, invasion, and metastasis of osteosarcoma. Methods: MiR-133a expression in human osteosarcoma cell lines and human normal osteoblastic cell line hFOB was investigated by real-time PCR (RT-PCR). The role of miR-133a in human osteosarcoma growth and invasion was assessed in cell lines in vitro and in vivo. Then, luciferase reporter assay validated IGF-1R as a downstream and functional target of miR-133a, and functional studies revealed that the anti-tumor effect of miR-133a was probably due to targeting and repressing of IGF-1R expression. Results: MiR-133a was lower expressed in human osteosarcoma cell lines than human normal osteoblastic cell line hFOB and its effect on inhibiting proliferation, invasion and metastasis is mediated by its direct interaction with the IGF-1R. Furthermore, the tumour-suppressive function of miR-133a probably contributed to inhibiting the activation AKT and ERK signaling pathway. Conclusion: MiR-133a suppresses osteosarcoma progression and metastasis by targeting IGF-1R in human osteosarcoma cells, providing a novel candidate prognostic factor and a potential anti-metastasis therapeutic target in osteosarcoma. © 2016 The Author(s) Published by S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Introduction: Multiple sclerosis (MS) and osteoporosis (OP) affect a substantial proportion of the population. Accumulating evidence suggests that MS patients are at high risk for OP. We performed a meta-analysis to identify risk factors for lowered bone mineral density (BMD) in MS patients.Methods: We searched for articles within the Medline, Embase and Cochrane Library databases, published up to March 2014, pertaining to associations between MS and BMD. A total of 11 studies was included in the meta-analysis.Results: The analysis indicated that MS patients have reduced lumbar spine, femur neck, and hip BMD compared with healthy controls (lumbar spine, standardized mean difference (SMD) = −0.76, 95% CI: −1.07, −0.45; femur neck, SMD = −0.56, 95% CI: −0.84, −0.29; and hip, SMD = −0.62, 95% CI: −0.96, −0.29). Further subgroup analysis revealed that a disease duration of >7 years, total steroid dose during the disease of >15 g, and an Expanded Disability Status Scale (EDSS) score of > 3, increased the risk of reduced BMD in the lumbar spine and femoral neck, but not in the hip. Meta-regression analysis did not explain the heterogeneity in the clinical characteristics or outcome definitions.Conclusions: Our meta-analysis suggests that MS patients have reduced overall BMD compared with healthy controls. Furthermore, disease duration (>7 years), total steroid dose (>15 g), and EDSS score (>3) are risk factors for reduced BMD in MS patients. [ABSTRACT FROM AUTHOR]

Challenges of repairing injuries and damage to the cartilage still remain in orthopedics. The characteristics of cartilage structure, especially avascular, make it a limited capacity of self-renewal. Articular cartilage defect or damage result from various causes will lead to degenerative osteoarthritis (OA). Surgical treatment and non-surgical treatment can temporarily alleviate symptoms to some extent but can't fundamentally restore the normal structure and function of cartilage, and therefore give rise to progressive degeneration. Autologous or allogeneic cartilage transplantation has been employed to the treatment of osteoarthritis for years. Nevertheless, the major deficiency of cartilage grafting is the inability and insufficiency to repair large cartilage defect. Implants are also unable to integrate with native tissue well. Adipose-derived stem cells (ASCs) can be easily isolated from subcutaneous fat tissues and harvest as intact cell sheets containing extracellular matrix (ECM), intercellular connect, ion channel, growth factor receptors, nexin and other important cell surface proteins by means of temperature-responsive culture dish (TCD). A cell sheet can provide a large amount of extracellular matrix, fibronectin, and cells contributing to the integration of cartilage. Decellularized extracellular matrix (DECM) of cartilage debris with excellent cell affinity and signal transduction is capable of driving cartilage homeostasis and regeneration. Appropriate decellularization process would remove cellular remnants of cartilage debris, keep the mechanical properties, and avoid the adverse immune response of allografts effectively. Micro-porous cartilage debris conduces to cell migration and angiogenesis. The cell-round shape of adipose-derived stem cells cultured in the three-dimensional (3D) system provided by hydrogel is more susceptible to chondrogenic stimulation and prevents it from fibroblast-like phenotypic conversion. We hypothesize that adipose-derived stem cell sheet encapsulated construct of micro-porous decellularized cartilage debris and hydrogel can effectively promote regeneration of cartilage defect. The construct of decellularized cartilage debris and hydrogel provide a favorable microenvironment for stem cells. Adipose-derived stem cells sheet supply fibronectin, collagen, and cells contributing to integration and regeneration of cartilage restore. Moreover, the constructs can be shaped and fabricated according to the configuration of target defect, especially in osteoarthritis, which is promising for clinical application. [ABSTRACT FROM AUTHOR]

Poly(lactide-co-glycolide) (PLGA) scaffolds embedded spatially with hydroxyapatite (HA) particles on the pore walls (PLGA/HA-S) were fabricated by using HA-coated paraffin spheres as porogens, which were prepared by Pickering emulsion. For comparisons, PLGA scaffolds loaded with same amount of HA particles (2%) in the matrix (PLGA/HA-M) and pure PLGA scaffolds were prepared by using pure paraffin spheres as porogens. Although the three types of scaffolds had same pore size (450-600 µm) and similar porosity (90%-93%), the PLGA/HA-S showed the highest compression modulus. The embedment of the HA particles on the pore walls endow the PLGA/HA-S scaffold with a stronger ability of protein adsorption and mineralization as well as a larger mechanical strength against compression. In vitro culture of rat bone marrow stem cells revealed that cell morphology and proliferation ability were similar on all the scaffolds. However, the alkaline phosphatase activity was significantly improved for the cells cultured on the PLGA/HA-S scaffolds. Therefore, the method for fabricating scaffolds with spatially embedded nanoparticles provides a new way to obtain the bioactive scaffolds for tissue engineering. Copyright © 2011 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]

Abstract: The restoration of the degenerated articular cartilage in patients with osteoarthritis (OA) is still a challenge for researchers and clinicians. Drug interventions and surgical treatments have been widely attempted for cartilage regeneration in OA. However, the results were largely unsatisfactory. Autologous chondrocyte implantation (ACI) or matrix-induced autologous chondrocyte implantation (MACI) offers potential for the regeneration of cartilage over the long-term. However, due to the limitations and disadvantages of ACI, alternative therapies for cartilage regeneration are in need. The availability of large quantities of mesenchymal stem cells (MSCs) and the multilineage differentiation, especially their chondrogenic differentiation property, have made MSCs the most promising cell source for cartilage regeneration. In addition, MSCs have been shown the ability to undergo site-specific differentiation. MSCs can be obtained as MSC sheets using the temperature-responsive culture dish method. The MSC sheet can provide amounts of cells and extracellular matrix, which might provide the continuity between the implant and host cartilage, thus improving integrative cartilage repair. Moreover, OA is associated with progressive and often severe inflammation. MSCs not only have the ability to contribute structurally to tissue repair, but also possess potent immunomodulatory and anti-inflammatory effects. Taken together, these properties make MSC sheet promising candidate for cartilage repair in OA. We hypothesize that MSC sheet encapsulated cartilage debris can efficiently promote cartilage repair in OA patients. Chondrocytes can be obtained and cultured from small cartilage debris in vitro. Therefore, the chondrocytes may grow from the debris in cartilage defect and improve cartilage regeneration. MSC sheet provide amounts of cells, ECM and protein for cartilage regeneration and integration, and may play some roles of periosteum. The operation of MSC sheet encapsulated cartilage debris for cartilage repair is simple and practical. Moreover, the cell sheet/cartilage debris constructs can be easily shaped based on the size and shape of cartilage defects. The new method might have great potential in treating cartilage defects clinically, especially for OA patients. [Copyright &y& Elsevier]