Flexible Fabrication and Hybridization of Bioactive Hydrogels with Robust Osteogenic Potency.

Osteogenic scaffolds reproducing the natural bone composition, structures, and properties have represented the possible frontier of artificially orthopedic implants with the great potential to revolutionize surgical strategies against the bone-related diseases. However, it is difficult to achieve an all-in-one formula with the simultaneous requirement of favorable biocompatibility, flexible adhesion, high mechanical strength, and osteogenic effects. Here in this work, an osteogenic hydrogel scaffold fabricated by inorganic-in-organic integration between amine-modified bioactive glass (ABG) nanoparticles and poly(ethylene glycol) succinimidyl glutarate-polyethyleneimine (TSG-PEI) network was introduced as an all-in-one tool to flexibly adhere onto the defective tissue and subsequently accelerate the bone formation. Since the N-hydroxysuccinimide (NHS)-ester of tetra-PEG-SG polymer could quickly react with the NH2-abundant polyethyleneimine (PEI) polymer and ABG moieties, the TSG-PEI@ABG hydrogel was rapidly formed with tailorable structures and properties. Relying on the dense integration between the TSG-PEI network and ABG moieties on a nano-scale level, this hydrogel expressed powerful adhesion to tissue as well as durable stability for the engineered scaffolds. Therefore, its self-endowed biocompatibility, high adhesive strength, compressive modulus, and osteogenic potency enabled the prominent capacities on modulation of bone marrow mesenchymal stem cell (BMSCs) proliferation and differentiation, which may propose a potential strategy on the simultaneous scaffold fixation and bone regeneration promotion for the tissue engineering fields.

Single-cell RNA-seq analysis reveals that immune cells induce human nucleus pulposus ossification and degeneration.

Background and aims: Determining the transcriptomes and molecular mechanism underlying human degenerative nucleus pulposus (NP) is of critical importance for treating intervertebral disc degeneration (IDD). Here, we aimed to elucidate the detailed molecular mechanism of NP ossification and IDD using single-cell RNA sequencing. Methods: Single-cell RNA-seq and bioinformatic analysis were performed to identify NP cell populations with gene signatures, biological processes and pathways, and subpopulation analysis, RNA velocity analysis, and cell-to-cell communication analysis were performed in four IDD patients. We also verified the effects of immune cells on NP ossification using cultured NP cells and a well-established rat IDD model. Results: We identified five cell populations with gene expression profiles in degenerative NP at single-cell resolution. GO database analysis showed that degenerative NP-associated genes were mainly enriched in extracellular matrix organization, immune response, and ossification. Gene set enrichment analysis showed that rheumatoid arthritis signaling, antigen processing and presentation signaling were activated in the blood cell cluster. We revealed that stromal cells, which are progenitor cells, differentiated toward an ossification phenotype and delineated interactions between immune cells (macrophages and T cells) and stromal cells. Immune factors such as TNF-α, CD74 and CCL-3 promoted the differentiation of stromal cells toward an ossification phenotype in vitro. Blocking TNF-α with a specific inhibitor successfully reversed NP ossification and modified NP morphology in vivo. Conclusion: Our study revealed an increase in macrophages and T cells in degenerative NP, which induced stromal cell differentiation toward an ossification phenotype, and contributed to the identification of a novel therapeutic target to delay IDD.

Detection of Subchondral Bone Microcirculatory Perfusion in Adults with Early Osteonecrosis of the Femoral Head Using Contrast-Enhanced Ultrasound: A Prospective Study.

The aim of this study was to quantitatively assess subchondral bone microcirculation perfusion in adults with early osteonecrosis of the femoral head (ONFH) using contrast-enhanced ultrasound (CEUS) and to evaluate its correlation with the Association Research Circulation Osseous (ARCO) stage. We investigated 97 adult patients with definite ONFH by imaging a total of 155 hips, performing CEUS, storing images of CEUS processes at different ARCO stages and generating CEUS time-intensity curves (TICs) to obtain perfusion parameters. Differences in CEUS parameters at different ARCO stages were analyzed, and correlations were explored. A logistic regression model was constructed by incorporating the meaningful CEUS indicators. The CEUS parameters time to peak (TTP), peak intensity (PI), enhanced intensity (EI), ascending slope (AS), descending slope (DS) and area under the receiver operating characteristic curve (AUC) were significantly different in ARCO stage Ⅰ compared with stage ⅢA, and the same results were obtained in stage Ⅱ compared with stage ⅢA. However, there were no significant differences between stages Ⅰ and Ⅱ. The MTT (mean transit time) assay was not significantly different between the different stages. The receiver operating characteristic curve analysis of TTP, PI, EI, AS, DS and AUC in stages Ⅰ and ⅢA had a certain diagnostic efficacy, similar to the results in stages Ⅱ and ⅢA. The diagnostic performance of DS was less accurate in stages Ⅰ and ⅢA, while the diagnostic performance of TTP was less accurate in stages Ⅱ and ⅢA. ARCO stage was independently and negatively correlated with TTP and DS and independently and positively correlated with PI, EI, AS and AUC. The MTT assay was not correlated with ARCO stage. Logistic regression models containing statistically significant TTP, EI and AUC values were constructed, and all three values were closely related to the ARCO stage. In patients with different ARCO stages of ONFH, CEUS can effectively assess subchondral bone perfusion of the femoral head and is expected to become an effective imaging method for the diagnosis of early ONFH.

Pretreatment Computed Tomography-Based Machine Learning Models to Predict Outcomes in Hepatocellular Carcinoma Patients who Received Combined Treatment of Trans-Arterial Chemoembolization and Tyrosine Kinase Inhibitor.

Aim: Trans-arterial chemoembolization (TACE) in combination with tyrosine kinase inhibitor (TKI) has been evidenced to improve outcomes in a portion of patients with hepatocellular carcinoma (HCC). Developing biomarkers to identify patients who might benefit from the combined treatment is needed. This study aims to investigate the efficacy of radiomics/deep learning features-based models in predicting short-term disease control and overall survival (OS) in HCC patients who received the combined treatment. Materials and Methods: A total of 103 HCC patients who received the combined treatment from Sep. 2015 to Dec. 2019 were enrolled in the study. We exacted radiomics features and deep learning features of six pre-trained convolutional neural networks (CNNs) from pretreatment computed tomography (CT) images. The robustness of features was evaluated, and those with excellent stability were used to construct predictive models by combining each of the seven feature exactors, 13 feature selection methods and 12 classifiers. The models were evaluated for predicting short-term disease by using the area under the receiver operating characteristics curve (AUC) and relative standard deviation (RSD). The optimal models were further analyzed for predictive performance on overall survival. Results: A total of the 1,092 models (156 with radiomics features and 936 with deep learning features) were constructed. Radiomics_GINI_Nearest Neighbors (RGNN) and Resnet50_MIM_Nearest Neighbors (RMNN) were identified as optimal models, with the AUC of 0.87 and 0.94, accuracy of 0.89 and 0.92, sensitivity of 0.88 and 0.97, specificity of 0.90 and 0.90, precision of 0.87 and 0.83, F1 score of 0.89 and 0.92, and RSD of 1.30 and 0.26, respectively. Kaplan-Meier survival analysis showed that RGNN and RMNN were associated with better OS (p = 0.006 for RGNN and p = 0.033 for RMNN). Conclusion: Pretreatment CT-based radiomics/deep learning models could non-invasively and efficiently predict outcomes in HCC patients who received combined therapy of TACE and TKI.

Cortical bone trajectory screws in the treatment of lumbar degenerative disc disease in patients with osteoporosis.

Lumbar degenerative disc disease (DDD) in the elderly population remains a global health problem, especially in patients with osteoporosis. Osteoporosis in the elderly can cause failure of internal fixation. Cortical bone trajectory (CBT) is an effective, safe and minimally invasive technique for the treatment of lumbar DDD in patients with osteoporosis. In this review, we analyzed the anatomy, biomechanics, and advantages of the CBT technique in lumbar DDD and revision surgery. Additionally, the clinical trials and case reports, indications, advancements and limitations of this technique were further discussed and reviewed. Finally, we concluded that the CBT technique can be a practical, effective and safe alternative to traditional pedicle screw fixation, especially in DDD patients with osteoporosis.

A Novel Rat Tail Needle Minimally Invasive Puncture Model Using Three-Dimensional Printing for Disk Degeneration and Progressive Osteogenesis Research.

Many studies focused on the annulus fibrosus (AF) injury in rodent tail model for the intervertebral disk degeneration (IDD) research. However, previous studies caused tremendous injury of intervertebral disk (IVD) by penetrating whole disk. This study aimed to build a progressive IDD rodent tail model by a novel device for precise and minimally invasive puncture in AF. A precise puncture device was customized by 3D Printing Technique. 40 rodent tail IVDs were randomly grouped as follows: group A, non-puncture; group B, annulus needle puncture (ANP) for 4 week; group C, ANP for 8 week; and group D, ANP for 12 week. Pre- and post-puncture IVD height on radiographs and IVD signal intensity on T2 magnetic resonance imaging (MRI) were measured. Average bone density (ABD) on the end of coccygeal vertebrae between punctured disk was measured on the radiographs. Hematoxylin and eosin, TUNEL staining methods, immunofluorescence for cleaved-caspas3 and immunohistochemistry for aggrecan and collagen II were performed. Progressively and significantly increasing IVD height loss and degenerative grade were observed following the time points. The ABD was respectively, 81.20 ± 4.63 in group A, 83.93±3.18 in group B, 92.65 ± 4.32 in group C, 98.87 ± 6.69 in group D. In both group C and group D, there were significant differences with group A. In histology, increasing number of AF cells was noted in group B. In both group C and D, the fissures in AF were obviously observed, and a marked reduction of AF cells were also observed. In all ANP groups, there were significant decrease in number of NP cells, as well as aggrecan and collagen II contents. TUNEL assay showed cellular apoptosis were stimulated in all puncture group, especially in group D. A progressive IDD rat model could be standardly established by the micro-injury IVD puncture using a novel 3D printing device. This animal model provided a potential application for research of progressive hyperosteogeny following IDD development.

A self-amplifying loop of YAP and SHH drives formation and expansion of heterotopic ossification.

Heterotopic ossification (HO) occurs as a common complication after injury or in genetic disorders. The mechanisms underlying HO remain incompletely understood, and there are no approved prophylactic or secondary treatments available. Here, we identify a self-amplifying, self-propagating loop of Yes-associated protein (YAP)-Sonic hedgehog (SHH) as a core molecular mechanism underlying diverse forms of HO. In mouse models of progressive osseous heteroplasia (POH), a disease caused by null mutations in GNAS, we found that Gnas-/- mesenchymal cells secreted SHH, which induced osteoblast differentiation of the surrounding wild-type cells. We further showed that loss of Gnas led to activation of YAP transcription activity, which directly drove Shh expression. Secreted SHH further induced YAP activation, Shh expression, and osteoblast differentiation in surrounding wild-type cells. This self-propagating positive feedback loop was both necessary and sufficient for HO expansion and could act independently of Gnas in fibrodysplasia ossificans progressiva (FOP), another genetic HO, and nonhereditary HO mouse models. Genetic or pharmacological inhibition of YAP or SHH abolished HO in POH and FOP and acquired HO mouse models without affecting normal bone homeostasis, providing a previously unrecognized therapeutic rationale to prevent, reduce, and shrink HO.

Analysis and improvement of the three-column spinal theory.

BACKGROUND: Denis and Ferguson et al.'s three-column spinal theory has been widely accepted and applied. However, this three-column theory was proposed based solely on observation and experience without thorough documented data and analysis. The aim of this study was to analyze and improve Denis and Ferguson et al.'s three-column spinal theory to propose a novel three-column concept in epidemiology, morphology and biomechanics. METHODS: A retrospective analysis of the computed tomography imaging data of patients with a diagnosis of T11-L5 vertebral fractures was conducted between February 2010 and December 2018. Three-dimensional (3D) distribution maps of fracture lines of all subjects were obtained based on 3D mapping techniques. In addition, a 25-year-old health male volunteer was recruited for the vertebral finite element force analysis. RESULTS: The present study enrolled 459 patients (age: 48 ± 11.42 years), containing a total of 521 fractured vertebrae. The fracture lines peaked in the upper and the outer third sections of the vertebra, starting from the anterior part of the vertebral pedicles in 3-D maps. Regarding flexion and extension of the spine, the last third of the vertebral body in front of the spinal canal was one main stress center in the finite element analysis. The stress on the vertebral body was greater in front of the pedicles in the lateral bending. CONCLUSION: The study reveals that the posterior one-third of the vertebral body in front of the spinal canal and the posterior one-third of the vertebral body in front of the pedicle are very different in terms of fracture characteristics and risks to spinal canal (3D maps and stress distributing graphs), therefore, they should be classified as different columns. We provide strong evidence that Su's three-column theory complies with the characteristics of vertebral physiological structure, vertebral fracture, and vertebral biomechanics.

Bioinformatics Analysis of the Molecular Mechanism of Late-Stage Heterotopic Ossification.

BACKGROUND: Heterotopic ossification (HO) is a common disease happened in soft tissues after injury. The present study utilized the bioinformatics method to analyze the HO samples in a mouse burn/tenotomy-induced HO model to identify the possible key points and treatment targets. METHODS: The transcriptome profiles of GSE126118 were obtained from the Gene Expression Omnibus (GEO) database. The study was based on a mouse burn/tenotomy-induced HO model, and 2 tenotomy samples and 3 uninjured contralateral hindlimb tendon samples were collected at 3 weeks after injury for further analysis. The transcripts per million approach was performed for background correction and normalization; then, the differentially expressed genes (DEGs) were detected using the limma R package with the settings p < 0.01 and ∣log2FC∣ > 2.0. The Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment, and the protein-protein interaction (PPI) network analysis were performed with the detected DEGs. RESULTS: A total of 74 DEGs were upregulated, and 159 DEGs were downregulated between the tenotomy and uninjured tendon group. Pathway and process enrichment analyses demonstrated that the upregulated DEGs were mainly associated with terms related to ECM remodeling, ossification, angiogenesis, inflammation, etc., and the downregulated DEGs were mainly associated with oxidative phosphorylation, metabolic process, etc. CONCLUSION: The results of GO, KEGG, and PPI network analyses suggested that the ECM remodeling, ossification, angiogenesis, and inflammation processes were markedly upregulated in the tenotomy site. And the oxidative phosphorylation and metabolic processes were markedly downregulated. These findings provide valuable clues for highlighting the characteristics of late-stage HO and investigating possible treatments.

ßig-h3 enhances chondrogenesis via promoting mesenchymal condensation in rat Achilles tendon heterotopic ossification model.

Heterotopic ossification (HO) is a poorly characterized disease with ectopic bone formation in the musculoskeletal soft tissues. HO is widely considered as a tissue repair process goes away, with endochondral ossification to be the major pathological basis. The molecular mechanism of how the resident/recruited progenitor cells for tissue regeneration error differentiated into the chondrocytes remains unknown. Here, we found Transforming Growth Factor B Induced Gene Human Clone 3 (ßig-h3) was highly expressed in the inflammation and chondrogenesis stages of a heterotopic ossification model after rat Achilles tendon injury, as well as upon chondrogenic differentiation conditions in vitro. ßig-h3 functioned as an extracellular matrix protein, which was induced by TGFß signaling, could bind to the injured tendon-derived stem cells (iTDSCs) and inhibit the attachment of iTDSCs to collagen I. Exogenous ßig-h3 was also found able to accelerate the process of mesenchymal condensation of cultured iTDSCs and promote chondrogenic differentiation in vitro, and additional injection of iTDSCs could promote endochondral ossification in Achilles tendon injury model. Taken together, ßig-h3 might function as an adhesion protein that inhibited the attachment of iTDSCs to collagen I (the injury site) but promoted the attachment of iTDSCs to each other, which resulted in promoting chondrogenic differentiation.

Three-dimensional computed tomography mapping of posterior malleolar fractures.

BACKGROUND: Posterior malleolar fractures have been reported to occur in < 40% of ankle fractures. AIM: To reveal the recurrent patterns and characteristics of posterior malleolar fractures by creating fracture maps of the posterior malleolar fractures through the use of computed tomography mapping. METHODS: A consecutive series of posterior malleolar fractures was used to create three-dimensional reconstruction images, which were oriented and superimposed to fit an ankle model template by both aligning specific biolandmarks and reducing reconstructed fracture fragments. Fracture lines were found and traced in order to generate an ankle fracture map. RESULTS: This study involved 112 patients with a mean age of 49, comprising 32 pronation-external rotation grade IV fractures and 80 supination-external rotation grade IV fractures according to the Lauge-Hansen classification system. Three-dimensional maps showed that the posterior ankle fracture fragments in the supination-external rotation grade IV group were relatively smaller than those in the pronation-external rotation grade IV group after posterior malleolus fracture. In addition, the distribution analyses on posterior malleolus fracture lines indicated that the supination-external rotation grade IV group tended to have higher linear density but more concentrated and orderly distribution fractures compared to the pronation-external rotation grade IV group. CONCLUSION: Fracture maps revealed the fracture characteristics and recurrent patterns of posterior malleolar fractures, which might help to improve the understanding of ankle fracture as well as increase opportunities for follow-up research and aid clinical decision-making.

A novel tissue separation method for determining upper trunk center of gravity in patients with thoracolumbar kyphosis using MIMICS.

BACKGROUND: In the past, a variety of methods have been explored to determine the center of gravity or gravity line, such as the hanging method or force plate analysis. However, these methods possess limitations related to precise location of center of gravity. The aim of this study was to describe a tissue separation method to precisely calculate the center of gravity in upper trunk. METHODS: Twenty post-operative patients with thoracolumbar kyphosis were retrospectively studied. Center of gravity models were computed: T1-T5 segment, T6-T10 segment and T1-T10 segment. The tissue separation method was used to calculate the center of gravity in each segment. A new center of gravity was composited from T1-T5 segment and T6-T10 segment by composition formula. Similarity and collinearity between center of gravity models was analyzed to verify the reliability of tissue separation method. Correlation between gravity line and theoretic hip axis was compared on pre- and post-operative radiographs to explore their potential application for surgical plan. FINDINGS: Composited center of gravity had significant correlation and high similarity with center of gravity in T1-T10 segment. There was high collinearity between center of gravity points. The post-operative included angle between the gravity line and theoretic hip axis significantly decreased to nearly 0°. INTERPRETATION: These findings demonstrate the MIMICS can reliably calculate the center of gravity in the upper trunk by a tissue separation method. The pre-operative included angle between the gravity line and theoretic hip axis was suggested as a parameter to improve surgical design for thoracolumbar kyphosis correction.

Melatonin modulates IL-1ß-induced extracellular matrix remodeling in human nucleus pulposus cells and attenuates rat intervertebral disc degeneration and inflammation.

The inflammatory-associated factors interleukin-1ß (IL-1ß), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) are widely reported to be associated with intervertebral disc (IVD) degeneration (IVDD). N-acetyl-5-methoxytryptamine (melatonin) is a natural hormone secreted by the pineal gland which has been shown to participate in several physiological and pathological progresses, such as aging, anti-inflammation, anti-apoptosis and autophagy regulation. However, the effects of melatonin on IVD remain unclear. In the present study, we treated human nucleus pulposus cells (NPCs) with melatonin and discovered that melatonin could modulate extracellular matrix (ECM) remodeling induced by IL-1ß by enhancing collagen II and aggrecan expression levels and by downregulating matrix metalloproteinase-3 (MMP-3) levels. These findings were verified by western blot and immunofluorescence assays. Intraperitoneal injection of melatonin mitigated IVDD in the rat tail puncture model. X-ray and magnetic resonance imaging (MRI), as well as hematoxylin-eosin (H&E), Safranine O-Green, Alcian blue and Celium red staining methods were adopted to evaluate IVDD grades, the structural integrity of nucleus pulposus (NP) and annulus fibrosus (AF) and the damage and calcification of the cartilage endplate. Melatonin reduced inflammatory cell aggregation and the release of the inflammatory factors IL-1ß, IL-6, TNF-α as determined by immunohistochemistry. In conclusion, the present study demonstrated that melatonin could modulate ECM remodeling by IL-1ß in vitro and attenuate the IVDD and induction of inflammation in a rat tail puncture model in vivo. The data demonstrated that melatonin may contribute to the restoration processs of IVD following damage and may be used as a potential novel therapy for IVDD.

3D Computed Tomography Mapping of Thoracolumbar Vertebrae Fractures.

BACKGROUND Fractures of the thoracolumbar (TL) spine represent 90% of all spinal fractures, followed by cervical and lumbar spine fractures. This study aimed to create fracture maps of the traumatic thoracolumbar (TL) fracture vertebral body (T12-L2) through the use of CT mapping as a big data visualization method to reveal recurrent patterns and characteristics of traumatic TL fractures. MATERIAL AND METHODS A consecutive series of 174 fractured vertebrae (T12-L2) was used to create three-dimensional (3D) reconstruction images, which were superimposed and oriented to fit a model vertebral template by aligning specific bio-landmarks and reducing reconstructed fracture fragments. Fracture lines were found and traced to create a fracture map of the vertebral body. RESULTS Our study consisted of 165 patients with an average age of 47 years. A total of 174 fractured vertebrae were collected, consisting of 59 T12 vertebral fractures, 60 L1 vertebral fractures, and 55 L2 vertebral fractures. Two-dimensional (2D) maps, 3D maps, and heat maps showed that the fracture lines tended to be concentrated in the upper third and anterior third of the vertebral body, as well as being distributed in annular wedges along the anterior and lateral sides of the vertebral body. When compared with T12, the distribution of fracture lines in L1 and especially in L2 was more scattered and disorganized. CONCLUSIONS Fracture maps revealed recurrent patterns and characteristics of the traumatic TL fracture vertebral body, which improves understanding of TL fractures, as well as helping to increase opportunities for follow-up research and aid clinical decision-making.

Percutaneous Pedicle Screw Fixation Alone Versus Debridement and Fusion Surgery for the Treatment of Early Spinal Tuberculosis: A Retrospective Cohort Study.

BACKGROUND Advances in diagnostic imaging techniques make it possible to detect tuberculosis (TB) lesions earlier, when only bone destruction or inflammatory infiltration is demonstrated. These techniques provide doctors with more opportunities to treat TB in the early stages of the disease. Traditional aggressive debridement surgery increases the risk of surgical complications. Therefore, we aimed to determine whether using percutaneous pedicle screw (PPS) fixation alone for the treatment of early spinal TB was a valid and less invasive surgical technique. MATERIAL AND METHODS We retrospectively reviewed the clinical and radiographic outcomes in cases with thoracic or lumbar TB treated with PPS surgery or hybrid surgery between January 2010 and January 2017. The operative time, blood loss, length of hospital stay, and hospitalization costs in the 2 groups were recorded and compared. Erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) before and at 18 months after surgery were tested to evaluate TB progress. Back pain was measured using the visual analog scale (VAS) before the operation and at the final follow-up. Radiological outcomes were evaluated at 1, 6, 12, and 18 months after surgery. A paired t-test was used to evaluate preoperative and postoperative clinical outcomes using SPSS 19.0 software. P values less than 0.05 were considered to be significant. RESULTS A total of 42 patients were involved in this retrospective study. In both groups, the average preoperative ESR, CRP level, and VAS score for back pain significantly decreased after surgery. In the PPS group, the operative time, blood loss, hospital stay, and hospitalization costs were all significantly lower than those in the hybrid group. X-ray and CT images showed satisfactory bone fusion and good maintenance of spinal alignment in both groups at the final follow-up. CONCLUSIONS PPS fixation alone was a valid and less invasive surgery for the treatment of early spinal TB. Furthermore, the recovery process of spinal TB can be facilitated using a "simple" internal fixation procedure, and bone fusion can be achieved without aggressive debridement and bone graft surgery.

Oxidative damage induces apoptosis and promotes calcification in disc cartilage endplate cell through ROS/MAPK/NF-κB pathway: Implications for disc degeneration.

Cartilage endplate (CEP) cell calcification and apoptosis play a vital role in the intervertebral disc degeneration (IVDD). Oxidative stress is a key factor in inducing programmed cell death and cartilage calcification. However, the cell death and calcification of cartilage endplate cells under oxidative stress have never been described. The present study investigated the apoptosis and calcification in the cartilage endplate cell under oxidative stress induced by H2O2 to understand the underlying mechanism of IVDD. The cartilage endplate cells isolated from human lumbar discs were subjected to different concentrations of H2O2 for various time periods. The cell viability was determined by CCK-8 assay, whereas Western blot, immunofluorescence, and Alcian blue, Alizarin red, and Von Kossa staining evaluated the apoptosis and calcification. The level of mitochondria-specific reactive oxygen species (ROS) was quantified with an oxygen radical-sensitive probe-MitoSOX. The potential signaling pathways were investigated by Western blot after the addition of N-acetyl-l-cysteine (NAC). We found that the oxidative stress induced by H2O2 increased the apoptosis and subsequently the calcification in the cartilage endplate cells through the ROS/p38/ERK/p65 pathway. The apoptosis and the calcification of the cartilage endplate cells induced by H2O2 can be abolished by NAC. These results suggested that regulating the apoptosis and the calcification in the cartilage endplate cells under oxidative stress should be advantageous for the survival of cells and might delay the process of disc degeneration.

High prevalence of PCV2d in Hunan province, China: a retrospective analysis of samples collected from 2006 to 2016.

Porcine circovirus 2 (PCV2) has been widely prevailing in China since the first report in 2001, causing huge economic losses to the pig industry. In the present study, 674 samples were collected from 2006 to 2016 in Hunan province, and 62% were positive for PCV2. An increase was observed from 2006 to 2011 (72.1%-89.1%), and a decrease was observed from 2012 to 2016 (78.9%-36.8%). The prevalence of genotype PCV2a, PCV2b, and PCV2d was 0, 44.7% and 67%, respectively. During 2006-2007, PCV2b was the main genotype circulating in Hunan, while, in 2008, PCV2d became the predominant one. Coinfection with PCV2b and PCV2d was observed frequently, and the positive rates of coinfection ranged from 6.3% to 18.9% during 2006-2016. The complete genome was sequenced for 54 positive samples, and four were identified as PCV2b-1, 22 as PCV2b-2, four as PCV2d-1 and 24 as PCV2d-2, based on phylogenetic analysis of the complete genome and ORF2 region. Recombination analysis using the complete genome sequences of these isolates revealed a high recombination rate of 27.7% (17/54), and showed that recombination occurred mainly in the ORF1 region. This shows that the prevalence of PCV2 has clearly decreased in recent years and that PCV2d has become a predominant genotype since 2008. In addition, frequent recombination events were observed in the PCV2 isolates from Hunan, China.

The Biomechanical Study of Extraforaminal Lumbar Interbody Fusion: A Three-Dimensional Finite-Element Analysis.

OBJECTIVE: Finite-element method was used to evaluate biomechanics stability of extraforaminal lumbar interbody fusion (ELIF) under different internal fixation. METHODS: The L3-L5 level finite-element model was established to simulate decompression and internal fixation at L4-L5 segment. The intact finite model was treated in accordance with the different internal fixation. The treatment groups were exerted 400 N load and 6 N·m additional force from motion to calculate the angular displacement of L4-L5. RESULTS: The ROMs were smaller in all internal fixation groups than those in the intact model. Furthermore, the ROMs were smaller in ELIF + UPS group than in TLIF + UPS group under all operating conditions, especially left lateral flexion and right rotation. The ROMs were higher in ELIF + UPS group than in TLIF + BPS group. The ROMs of ELIF + UPS + TLFS group were much smaller than those in ELIF + UPS group, and as compared with TLIF + BPS group, there was no significant difference in the range of experimental loading. DISCUSSION: The biomechanical stability of ELIF with unilateral pedicle screw fixation is superior to that of TLIF with unilateral pedicle screw fixation but lower than that of TLIF with bilateral pedicle screws fixation. The stability of ELIF with unilateral fixation can be further improved by supplementing a translaminar facet screw.

Molecular characterization of Porcine sapelovirus in Hunan, China.

Porcine sapeloviruses (PSVs) are widely distributed in pig populations; however, little information on their evolutionary history and the mechanisms driving their divergence is available. Therefore, in the present study, 241 fecal samples and 91 intestinal contents collected from pigs at 26 farms in Hunan, China, were tested for the presence of PSVs. The overall PSV positivity rate was 46.39 %, with a particularly high infection rate detected in nursery and fattening pigs. A total of 29 PSV strains (PSV-HuNs) were isolated, with these showing high genetic diversity based on phylogenetic and pairwise distance analyses of the capsid-protein gene sequences. Incongruence between phylognetic trees of the capsid-protein and 3CD regions indicated frequent recombination within the PSV-HuNs, and a putative recombinant hotspot near the 3' end of the P1 region was identified. Our results suggested that recombination played an important role in driving PSV genetic diversity and evolution.