Morroniside ameliorates glucocorticoid-induced osteoporosis and promotes osteoblastogenesis by interacting with sodium-glucose cotransporter 2.

CONTEXT: Morroniside (MOR) possesses antiosteoporosis (OP) effects, but its molecular target and relevant mechanisms remain unknown. OBJECTIVE: We investigated the effects of MOR on glucocorticoid-induced OP and osteoblastogenesis and its underlying mechanisms. MATERIALS AND METHODS: The effects of MOR (10-100 µM) on the proliferation and differentiation of MC3T3-E1 cells were studied in vitro. The glucocorticoid-induced zebrafish OP model was treated with 10, 20 and 40 µM MOR for five days to evaluate its effects on vertebral bone density and related osteogenic markers. In addition, molecular targets prediction and molecular docking analysis were carried out to explore the binding interactions of MOR with the target proteins. RESULTS: In cultured MC3T3-E1 cells, 20 µM MOR significantly increased cell viability (1.64 ± 0.12 vs. 0.95 ± 0.16; p < 0.01) and cell differentiation (1.57 ± 0.01 vs. 1.00 ± 0.04; p < 0.01) compared to the control group. MOR treatment significantly ameliorated vertebral bone loss in the glucocorticoid-induced OP zebrafish model (0.86 ± 0.02 vs. 0.40 ± 0.03; p < 0.01) and restored the expression of osteoblast-specific markers, including ALP, Runx2 and Col-І. Ligand-based target prediction and molecular docking revealed the binding interaction between MOR and the glucose pockets in sodium-glucose cotransporter 2 (SGLT2). DISCUSSION AND CONCLUSIONS: These findings demonstrated that MOR treatment promoted osteoblastogenesis and ameliorated glucocorticoid-induced OP by targeting SGLT2, which may provide therapeutic potential in managing glucocorticoid-induced OP.

Ginsenoside Rb1 Facilitates Browning by Repressing Wnt/ß-Catenin Signaling in 3T3-L1 Adipocytes.

BACKGROUND The discovery of browning in white adipose tissue has provided new ideas for treating obesity. Many studies have reported that ginsenoside Rb1 (G-Rb1) has activity against diabetes, inflammation, and obesity, but further investigation is needed on the effect and mechanism of G-Rb1 on browning. MATERIAL AND METHODS We treated 3T3-L1 adipocytes with 0-200 µM G-Rb1, and 0.5 µM Compound 3f and 30 µM SKL2001 were used to activate Wnt/b-catenin signaling. Adipocyte activity was evaluated by Cell Counting Kit-8. Oil Red O staining was used to detect the lipid droplets. Quantitative real-time polymerase chain reaction was used to measure the expression of Cd-137, Cited-1, Txb-1, Prdm-16, and Ucp-1 mRNA. Western blotting was used to measure the expression of Ucp-1, pGSK-3ß (Ser 9), GSK- 3ß, and ß-catenin proteins. The expression of Ucp-1 was also detected with immunofluorescence. RESULTS Adipocyte activity was not affected by 0-100 µM G-Rb1. However, G-Rb1 dose-dependently reduced the accumulation of lipid droplets; increased the expression of Cd-137, Cited-1, Txb-1, Prdm-16, and Ucp-1 mRNA; and increased the expression of Ucp-1, pGSK-3ß (Ser 9), GSK-3ß, and ß-catenin proteins. The accumulation of lipid droplets and the expression of Ucp-1 protein decreased as b-catenin increased. CONCLUSIONS G-Rb1 at various concentrations (0-100 µM) promoted the browning of adipocytes in a dose-dependent manner. Further, we confirmed that activation of Wnt/ß-catenin signaling could inhibit browning. Therefore, the browning promoted by G-Rb1 may be associated with the inhibition of Wnt/ß-catenin signaling.

miR-532-3p inhibits osteogenic differentiation in MC3T3-E1 cells by downregulating ETS1.

BACKGROUND: Many studies had identified that MicroRNAs (miRNAs) could affect bone metabolism by regulating the expression of various proteins. This study explored the effect and mechanism of miR-532-3p on osteogenic differentiation. METHODS: We analyzed the content of miR-532-3p in osteoporosis patients, osteoporosis rats, and osteogenic induced MC3T3-E1 cells. MiR-532-3p mimic or inhibitor utilized to alter intracellular miR-532-3p content. MTT method executed to detect the effect of miR-532-3p on osteoblast proliferation. Real-time qPCR, Western blot, alkaline phosphatase staining, and alizarin red staining utilized to ascertain the influence of miR-532-3p on osteogenic differentiation. Then, databases and a dual-luciferase reporter gene assay used to verify the target of miR-532-3p. Furthermore, the lentiviral vector was utilized to overexpress interesting target gene expression and checked whether the target gene was involved in the regulation of osteogenic differentiation by miR-532-3p. RESULTS: MiR-532-3p expression boosted in low bone mineral density (BMD) patients and rats. In MC3T3-E1 cells, miR-532-3p expression gradually decreased as osteogenic induction matures. MiR-532-3p mimic negatively regulated succinate dehydrogenase (SDH) activity, alkaline phosphatase (ALP) activity, mineralization ability, the osteogenic-associated gene (Col1A1, Runx2, ALP, OPN, and OCN) and E-26 transformation specific-1 (ETS1) expression of MC3T3-E1 cells. Things are the opposite of the miR-532-3p inhibitor. ETS1 identified as the miR-532-3p target gene, and miR-532-3p could inhibit its expression. Besides, improved ETS1 expression could rescue the suppressive effect of miR-532-3p mimic on osteogenic differentiation. CONCLUSION: miR-532-3p can suppress osteogenic differentiation by downregulating ETS1 expression.

Comparative clinical efficacy and safety of cortical bone trajectory screw fixation and traditional pedicle screw fixation in posterior lumbar fusion: a systematic review and meta-analysis.

PURPOSE: To compare the clinical efficacy and safety between cortical bone trajectory (CBT) and pedicle screw (PS) in posterior lumbar fusion surgery. METHODS: Five electronic databases were used to identify relevant studies comparing the clinical efficacy and safety between CBT and PS. The main outcomes were postoperative fusion rates and complication (especially in superior facet joint violations, symptomatic ASD, wound infection, dural tear, screw malposition and hematoma). The secondary results included operation time, intraoperative blood loss, length of hospital stay, incision length, ODI, VAS, JOA score, JOA recovery rate, patients' satisfaction and health-related quality of life. RESULTS: The outcomes showed that there was no significant difference in terms of fusion rate (p = 0.55), back and leg VAS score (p > 0.05), JOA score (p = 0.08) and incidence of reoperation (p = 0.07). However, CBT was superior to PS with Oswestry Disability Index (ODI) (p = 0.02), JOA recovery rate (p < 0.00001) and patients' satisfaction (p = 0.001). In addition, CBT was superior to PS with significantly lower incidence of superior facet joint violation and symptomatic ASD. However, there was no significant difference regarding wound infection (p > 0.05) and screw malposition (p > 0.05). CBT group required significant shorter operation time, less blood loss, shorter incision length and shorter length of hospital stay in comparison with PS group (p < 0.05). CONCLUSIONS: Both CBT and PS achieve similar, fusion rate and revision surgery rate. Furthermore, CBT is superior to PS with lower incidence of complications, shorter operation time, less blood loss, shorter incision length and shorter length of hospital stay. These slides can be retrieved under Electronic Supplementary Material.

Efficacy of antifibrinolytic agents on surgical bleeding and transfusion requirements in spine surgery: a meta-analysis.

PURPOSE: Spine surgery is usually associated with large amount of blood loss and blood transfusion. Excessive blood loss may cause hypotension, inadequate oxygenation of organs, necessitate allogeneic blood transfusion, and spinal epidural hematoma formation. Aprotinin, TXA, and EACA are antifibrinolytics currently offered as prophylactic agents to reduce surgery-associated blood loss. The purpose of this study was to assess the efficacy of using antifibrinolytic agents in reducing blood loss and blood transfusions in spine surgery. METHODS: PubMed, Embase, and Cochrane-controlled trials register were used to identify RCTs published before April 2015 that examined the effectiveness of intravenous aprotinin, tranexamic acid (TXA), and epsilon-aminocaproic acid (EACA) on reduction of blood loss and blood transfusions, compared with placebo in spine surgery. Randomized controlled trials reported the primary outcome that included total blood loss, intra-operative blood loss, post-operative blood loss, blood transfusion requirements, blood transfusion rate, and incidence of deep vein thrombosis. Meta-analysis was performed using the Stata12.0. Weighted mean difference with 95 % confidence intervals was used to summarize the findings across the trials for continuous outcomes. Dichotomous data were expressed as risk ratios with 95 % confidence intervals. A P < 0.05 was considered statistically significant. RESULTS: 17 studies involving 1191 patients were identified. Among them, 13 RCTs with 943 patients were included for the evaluation of total blood loss. Compared with the control group, the antifibrinolytic agents reduced total blood loss (SMD = -0.62; 95 % CI -0.75, -0.48; P = 0.000), The aprotinin group (SMD = -0.80; 95 % CI -1.22, -0.37; P = 0.938), The TXA group (SMD = -0.75; 95 % CI -0.93, -0.57; P = 0.000), and the EACA group (SMD = -0.28; 95 % CI -0.54, -0.01; P = 0.185). Thirteen RCTs with eight hundred and ninety four patients were included for the evaluation of intra-operative blood loss. Compared with the control group, the antifibrinolytic agents reduced intra-operative blood loss (SMD = -0.41; 95 % CI -0.55, -0.28; P = 0.010), The aprotinin group (SMD = -0.62; 95 % CI -0.93, -0.30; P = 0.862), The TXA group (SMD = -0.47; 95 % CI -0.64, -0.29; P = 0.005), and the EACA group (SMD = -0.16; 95 % CI -0.42, -0.11; P = 0.897). Eight RCTs with six hundred and seven patients were included for the evaluation of post-operative blood loss. Compared with the control group, the antifibrinolytic agents reduced post-operative blood loss (SMD = -0.68; 95 % CI -0.85, -0.51; P = 0.000), the aprotinin group (SMD = -0.48; 95 % CI -0.85, -0.12; P = 0.036), the TXA group (SMD = -0.80; 95 % CI -1.01, -0.59; P = 0.000), and the EACA group (SMD = -0.32; 95 % CI -0.68, -0.04; P = 0.009). Ten RCTs with seven hundred and twenty twopatients were included for the evaluation of blood transfusion. Compared with the control group, the antifibrinolytic agents reduced blood transfusion (SMD = -0.68; 95 % CI -0.85, -0.51; P = 0.000), the aprotinin group (SMD = -0.80; 95 % CI -1.22, -0.37; P = 0.938), the TXA group (SMD = -0.38; 95 % CI -0.58, -0.18; P = 0.000), and the EACA group (SMD = -0.28; 95 % CI -0.54, -0.01; P = 0.185). Twelve RCTs with eight hundred and fifteenpatients were included for the evaluation of blood transfusion rate. The transfusion rate was 35.6 % in the patients with antifibrinolytic agents and 55.2 % in the patients with placebo (RR = 0.75; 95 % CI 0.63, 0.89; P = 0.939). All studies were included for the evaluation of safety, with a total of eight thromboembolic events reported overall (two in the experimental group and six in the control group). CONCLUSIONS: The antifibrinolytic agents were able to reduce perioperative blood loss and transfusion requirements in spine surgery. TXA appeared more effective than aprotinin and EACA in reducing total blood loss, intra-operative blood loss, and blood transfusion according to the results of this analysis. The three groups in reducing the post-operative blood loss are significantly better than control groups. There was no evidence that the use of antifibrinolytic agents was a risk factor for thromboembolism in spine surgery. Further multicenter, large-sample, double-blind RCTs are required to confirm the efficacy and safety of the three antifibrinolytic agents in spine surgery.

Silencing of Transient Receptor Potential Channel 4 Alleviates oxLDL-induced Angiogenesis in Human Coronary Artery Endothelial Cells by Inhibition of VEGF and NF-κB.

BACKGROUND: Transient receptor potential channel 4 (TRPC4) plays central roles in endothelial cell function. The aim of this study was to investigate the silencing effects of TRPC4 on oxidized low-density lipoprotein (oxLDL)-induced angiogenesis in human coronary artery endothelial cells (HCAECs), as well as the underlying molecular mechanism involved in this process. MATERIAL/METHODS: HCAECs were transfected with small interfering RNA (siRNA) targeting TRPC4 (TRPC4-siRNA) or with a negative control (NC)-siRNA. The expression of TRPC4 was confirmed by real-time polymerase chain reaction (RT-PCR) and Western blotting. After the siRNA transfection, oxLDL was added to the medium. Cell proliferation, migration, and in vitro angiogenesis were determined by bromodeoxyuridine (BrdU) enzyme-linked immunosorbent assay (ELISA), Transwell assay and scratch-wound assay, respectively, and tube formation on Matrigel. Expression of vascular endothelial growth factor (VEGF) and nuclear factor (NF)-κB p65 were assessed by Western blotting. RESULTS: Both the mRNA and protein levels of TRPC4 were significantly reduced by transfection with TRPC4-siRNA compared to the control group or NC-siRNA group (P<0.05). Silencing of TRPC4 significantly decreased the cell proliferation, migration, and tube formation (all P<0.05). Furthermore, the expression levels of VEGF and NF-κB p65 were markedly lowered by silencing of TRPC4 in HCAECs. CONCLUSIONS: These results suggest that silencing of TRPC4 alleviates angiogenesis induced by oxLDL in HCAECs through inactivation of VEGF and NF-κB. Suppression of TRPC4 might be an alternative therapeutic strategy for atherosclerotic neovascularization.

Effect of lamina open angles in expansion open-door laminoplasty on the clinical results in treating cervical spondylotic myelopathy.

STUDY DESIGN: A retrospective study. OBJECTIVE: To evaluate and compare the relation of the efficacy and clinical results of expansion open-door laminoplasty (EOLP) with different angles in lamina open-door. SUMMARY OF BACKGROUND DATA: EOLP is currently the most widely adopted surgical treatment for cervical spondylotic myelopathy. Although many long-term clinical follow-up studies have reported that most patients recover satisfactorily after EOLP, there have been numerous reports regarding postoperative complications, such as stubborn axial symptoms (AS) and C5 palsy. The lamina open-door angles in EOLP play a decisive role in determining the openness of the door that affects clinical outcomes. Nonetheless, no thorough studies on different angles in EOLP have been published. METHOD: A total of 198 cervical spondylotic myelopathy patients who underwent posterior cervical EOLP and at least 24 months follow-up treatment between July 2006 and January 2009 were selected as case studies. Among the 198 cases used, there were 39 double-segment cases with the location being C3-C5 in 11 cases and C4-C6 in 28 cases, 97 three-segments (C4-C7) and 62 four segments (C3-C7). All of the patients underwent x-ray, computed tomography, and magnetic resonance imaging images for evaluation of the cervical spine. According to different opening angles, measured by computed tomography scan after operation 1 week, the patients were divided into 2 groups, group A (>30 degrees, 76 patients including 44 males and 32 females) and group B (15-30 degrees, 122 patients including 71 males and 51 females). All patients were followed up for over 24 months, clinical results including operative duration, intraoperative bleeding volume, postoperative complications, C2-C7 Cobb angle, cervical curvature index (CI), range of motion (ROM), and values after the spinal cord backward shift were analyzed statistically, evaluating the neurological function at final follow-up and calculating the improvement rate of nerve function recovery. RESULTS: There was no statistically significant difference (P>0.05) between the 2 groups in the following areas: the Japanese Orthopedic Association scores, C2-C7 Cobb angle, cervical CI, and ROM. In addition, operative duration and intraoperative bleeding volume between A group and B group showed no significant differences (P>0.05). After surgery, 51 patients (67.1%) in group A had AS, 8 patients (10.4%) had C5 palsy, and 1 patient had mild cervical kyphosis. Whereas postoperatively group B contained 37 cases (10.5%) with AS, 3 (2.4%) with C5 palsy, and in 4 cases (3.28%) the lamina open-door had reclosed. The rate of patients with AS and C5 palsy in group A was higher than group B. The incidence of postoperative complications between the 2 groups have a significant difference (P<0.05). The rate of improvement of Japanese Orthopedic Association scores in last follow-up between group A and group B did not reach statistical significance (P>0.05). At the 1-month follow-up the range of the value of spinal cord backward shift was 0-7.95 mm with the average being 2.41±0.46 mm. C2-C7 Cobb angle, CI, and ROM between the 2 groups revealed no statistical significance (P>0.05). ROM comparisons preoperatively and postoperatively between the 2 groups were significantly different (P<0.05). CONCLUSIONS: In different angles of lamina open-door, the improvement rate of neurological function after surgery had no statistically significant difference between 2 groups. When the open-door angle is maintained between 15 and 30 degrees, it can reduce the incidence of C5 palsy in the hinge side and AS, but we should prevent reclosure of the lamina open-door.

[The significance of lumbar MRI in the diagnosis of recurrented lumbar disc herniation after surgery].

OBJECTIVE: To retrospectively evaluate the clinical evaluation of preoperative lumbar T2 sagittal MRI image in predicting the recurrent lumbar disc herniation (RLDH). METHODS: Between January 2009 and April 2011, 28 patients were diagnosed as recurrent L4-5 disc herniation within 2.5 years after surgery, and 25 of them were included in the study as relapse group. At the same time, selected 25 patients implemented the same surgical methods in the same level as a control group randomly, they were all with good to excellent result and the follow-up time was at least 2.5 years. There was no statistical significance between the two groups in gender, age and body mass index(BMI) (P > 0.05). The lumbar MRI image of two groups of patients before surgery were collected and analyzed, with the disc degeneration grade classified. The χ(2) test was used to analyzed the difference of degeneration between the two groups of patients before surgery. Rank correlation analysis evaluated the correlation between disc degeneration and the period of time from the first operation to the recurrence. RESULTS: In terms of preoperative lumbar disc degeneration, there were 22 cases of low-grade disc degeneration and 3 cases of advanced disc degeneration in the relapse group and 5 cases and 20 cases respectively in the control group. there was significant difference between two groups (χ(2) = 23.27, P < 0.05), low-grade disc degeneration (gradesIand III) was significantly more frequent in the relapse group than in the control group. The patients with low-grade disc degeneration had a higher risk of recurrence, that was the risk of recurrent disc herniation increased by a factor of 4.4 from advanced disc degeneration to low-grade disc degeneration(OR = 4.4, 95%CI:1.983-9.765, P < 0.05). In cases of recurrence, the time interval between primary surgery and the recurrence of the patient with advanced disc degeneration was longer compared with low-grade disc degeneration (r = 0.733, P < 0.05). CONCLUSIONS: Preoperative lumbar MRI image may suggest the possibility of the recurrence lumbar disc herniation.Light disc degeneration is an important risk factor for recurrent disc herniation, and the time interval between primary surgery and the recurrence is positively correlated with severity of disc degeneration.

Up-regulation of NCAPG mediated by E2F1 facilitates the progression of osteosarcoma through the Wnt/ß-catenin signaling pathway.

Background: In recent years, there are few reports on non-SMC condensin I complex subunit G (NCAPG) in osteosarcoma. Our study aims to explore the biological role of NCAPG in osteosarcoma and its underlying molecular mechanism and to further clarify the reasons for the abnormal expression of NCAPG in osteosarcoma. Methods: Here, we mined The Cancer Genome Atlas (TCGA) Program public database through bioinformatics methods, analyzed the differential expression of NCAPG in sarcoma tissue and normal tissue, and explored the relationship between NCAPG expression level and sarcoma tissue differentiation, including tumor recurrence, metastasis, and patient survival. Next, the transcription factors responsible for the abnormal expression of NCAPG in osteosarcoma tumors were predicted by multiple online website tools and verified via cellular experiments. Subsequently, loss of function and cell phenotype experiments were performed to confirm the effect of NCAPG on the malignant biological behavior of osteosarcoma cells. Mechanistically, by reviewing the literature, we found that NCAPG can affect the malignant progression of many solid tumors by regulating the Wnt/ß-catenin signaling pathway. Therefore, we preliminarily investigated the potential effect of NCAPG on this pathway via western blot experiments in osteosarcoma. Results: Increased expression of NCAPG was found in sarcoma compared to normal tissues, which was positively correlated with poor differentiation, metastasis, and poor prognosis. Combining the transcription factor prediction results, correlation analysis, and expression level in the TCGA public database with validation outcomes of in vitro cell assays, we found that E2F transcription factor 1 (E2F1) regulated the increased expression of NCAPG in osteosarcoma. The results of cell phenotype experiments showed that silencing NCAPG could inhibit the proliferation, migration, and invasion of osteosarcoma cells. The preliminary mechanistic investigation suggested that NCAPG may affect osteosarcoma progression through the Wnt/ß-catenin pathway. Conclusions: Our data reveal that E2F1 facilitates NCAPG expression in osteosarcoma by regulating the transcription of the NCAPG gene. Up-regulation of NCAPG promotes osteosarcoma progression via the Wnt/ß-catenin signaling axis.

Enhancing osteoporosis treatment using a targeted, sustained-release drug delivery system based on macrocyclic amphiphile.

Osteoporosis, a prevalent systemic bone metabolic disorder, primarily affects postmenopausal women and is characterized by increased bone fragility and a heightened risk of fractures. The efficacy of current osteoporosis treatments is often limited by non-specific drug targeting and undesirable off-target skeletal side effects. To address this challenge, we have developed a novel hydroxyapatite-responsive drug delivery system. This system utilizes a self-assembled p-phosphonatocalix[4]arene tetradodecyl ether (PC4A12C), engineered to specifically target and sustain the release of osteoporosis medication at sites of bone remodeling. Our focus centers on icariin (ICA), a drug known for its potent osteogenic properties and minimal adverse effects. In vitro, ICA-loaded PC4A12C (ICA@PC4A12C) demonstrated enhanced proliferation, differentiation, and mineralization in bone marrow mesenchymal stem cells (BMSCs). In vivo, ICA@PC4A12C exhibited superior efficacy in specifically targeting bone tissue, ensuring a controlled and slow release of icariin directly within the bone environment. In an osteoporosis mouse model, treatment with ICA@PC4A12C showed notable enhancement in osteogenic activity and a significant increase in bone density compared to ICA alone. These results demonstrate the potential of PC4A12C as an effective drug carrier in the development of advanced antiosteoporotic drug delivery systems.

Supramolecular 3 in 1: A Lubrication and Co-Delivery System for Synergistic Advanced Osteoarthritis Therapy.

The complexity, heterogeneity, and drug resistance of diseases necessitate a shift in therapeutic paradigms from monotherapy to combination therapy, which could augment treatment efficiency. Effective treatment of advanced osteoarthritis (OA) requires addressing three key factors contributing to its deterioration: chronic joint inflammation, lubrication dysfunction, and cartilage-tissue degradation. Herein, we present a supramolecular nanomedicine of multifunctionality via molecular recognition and self-assembly. The employed macrocyclic carrier, zwitterion-modified cavitand (CV-2), not only accurately loads various drugs but also functions as a therapeutic agent with lubricating properties for the treatment of OA. Kartogenin (KGN), a drug for articular cartilage regeneration and protection, and flurbiprofen (FP), an anti-inflammatory agent, were coloaded onto CV-2 assembly, forming a supramolecular nanomedicine KGN&FP@CV-2. The three-in-one combination therapy of KGN&FP@CV-2 addresses the three pathological features for treating OA collectively, and thus provides long-term therapeutic benefits for OA through sustained drug release and intrinsic lubrication in vivo. The multifunctional integration of macrocyclic delivery and therapeutics provides a simple, flexible, and universal platform for the synergistic treatment of diseases involving multiple drugs.

Deficiency of Acetyltransferase nat10 in Zebrafish Causes Developmental Defects in the Visual Function.

Purpose: N4-acetylcytidine (ac4C) is a post-transcriptional RNA modification catalyzed by N-acetyltransferase 10 (NAT10), a critical factor known to influence mRNA stability. However, the role of ac4C in visual development remains unexplored. Methods: Analysis of public datasets and immunohistochemical staining were conducted to assess the expression pattern of nat10 in zebrafish. We used CRISPR/Cas9 and RNAi technologies to knockout (KO) and knockdown (KD) nat10, the zebrafish ortholog of human NAT10, and evaluated its effects on early development. To assess the impact of nat10 knockdown on visual function, we performed comprehensive histological evaluations and behavioral analyses. Transcriptome profiling and real-time (RT)-PCR were utilized to detect alterations in gene expression resulting from the nat10 knockdown. Dot-blot and RNA immunoprecipitation (RIP)-PCR analyses were conducted to verify changes in ac4C levels in both total RNA and opsin mRNA specifically. Additionally, we used the actinomycin D assay to examine the stability of opsin mRNA following the nat10 KD. Results: Our study found that the zebrafish NAT10 protein shares similar structural properties with its human counterpart. We observed that the nat10 gene was prominently expressed in the visual system during early zebrafish development. A deficiency of nat10 in zebrafish embryos resulted in increased mortality and developmental abnormalities. Behavioral and histological assessments indicated significant vision impairment in nat10 KD zebrafish. Transcriptomic analysis and RT-PCR identified substantial downregulation of retinal transcripts related to phototransduction, light response, photoreceptors, and visual perception in the nat10 KD group. Dot-blot and RIP-PCR analyses confirmed a pronounced reduction in ac4C levels in both total RNA and specifically in opsin messenger RNA (mRNA). Additionally, by evaluating mRNA decay in zebrafish treated with actinomycin D, we observed a significant decrease in the stability of opsin mRNA in the nat10 KD group. Conclusions: The ac4C-mediated mRNA modification plays an essential role in maintaining visual development and retinal function. The loss of NAT10-mediated ac4C modification results in significant disruptions to these processes, underlining the importance of this RNA modification in ocular development.

Predictors of residual low back pain in patients with osteoporotic vertebral fractures following percutaneous kyphoplasty.

Objective: Patients with osteoporotic vertebral fractures (OVFs) often suffer from residual low back pain (LBP) after percutaneous kyphoplasty (PKP). The purpose of this study was to identify risk factors for postoperative residual LBP and to develop a nomogram to predict the occurrence of residual LBP. Methods: We retrospectively reviewed 236 patients who underwent PKP for OVFs and had a minimum follow-up of 12 months. The mean age was 72.1 ± 6.3, 74.3% were female and 25.7% were male. Patients with LBP VAS scores ≥ 3.5 at the 12th month postoperatively were considered to have residual LBP. Risk factors for residual LBP were identified by univariate and multifactorial logistic regression analysis. Then, a predictive nomogram was constructed and validated using the bootstrap method. The discrimination, calibration, and clinical utility of the nomogram were assessed using a receiver operating characteristic curve (ROC), a calibration curve, and a decision curve analysis (DCA). Results: univariate and multifactorial logistic regression analysis identified depression (P = 0.02), intravertebral vacuum cleft (P = 0.01), no anti-osteoporosis treatment (P < 0.001), cement volume <3 ml (P = 0.02), and cement distrubution (P = 0.01) as independent risk factors for residual LBP. The area under the ROC was 0.83 (0.74-0.93) and further validated by bootstrap method was 0.83 (0.73-0.92). The calibration curve illustrated the consistency between the predicted probability and the observed results. DCA showed that nomogram exhibits clinical utility and net benefit when the threshold probability is between 6% and 73%. Conclusions: Our study found that depression, intravertebral vacuum cleft, no anti-osteoporosis treatment, cement volume <3 ml and cement distribution represent independent risk factors for residual LBP. The nomogram containing the above five predictors can accurately predict the risk of residual LBP after surgery.

Long noncoding RNA HCG18 Promotes Extracellular Matrix Degradation of Nucleus Pulposus Cells in Intervertebral Disc Degeneration by Regulating the miR-4306/EPAS1 Axis.

BACKGROUND: Intervertebral disc degeneration is a very common disease worldwide and the leading cause of low back pain. Long noncoding RNAs are novel players in intervertebral disc degeneration and have multiple functions. This study explored the role of long noncoding RNA HCG18 in regulating extracellular matrix (ECM) degradation in nucleus pulposus cells (NPCs) during intervertebral disc degeneration. METHODS: NPCs were subjected to interleukin-1ß to induce a degenerative model of NPCs. Cell viability was assessed using Cell Counting Kit-8 assay. Messenger RNA and protein expressions were examined by real-time quantitative polymerase chain reaction and Western blot. The location of HCG18 was determined by nucleocytoplasmic separation assay. The binding relationships between HCG18, MIR4306, and EPAS1 were verified by dual luciferase reporter gene assay and/or RNA immunoprecipitation assay. RESULTS: HCG18 was highly expressed in interleukin-1ß-induced degenerated NPCs, which was associated with reduced collagen II and aggrecan expression and increased MMP-13 and ADAMTS-4 expression. HCG18 knockdown could remarkably inhibit ECM degradation in IL-1ß-induced degenerated NPCs, while HCG18 overexpression had the opposite effect. Our molecular study further revealed that HCG18 could sponge MIR4306, and HCG18 knockdown could suppress ECM degradation in degenerated NPCs by elevating MIR4306 expression. In addition, EPAS1 was identified as the direct target of MIR4306. As expected, MIR4306 overexpression inhibited ECM degradation in degenerated NPCs by downregulating EPAS1. CONCLUSIONS: HCG18 promoted ECM degradation in degenerated NPCs via regulation of the MIR4306/EPAS1 axis.

Comparison of discover cervical disc arthroplasty and anterior cervical discectomy and fusion for the treatment of cervical degenerative disc diseases: A meta-analysis of prospective, randomized controlled trials.

Objective: This study aims to evaluate the clinical efficacy and safety between Discover cervical disc arthroplasty (DCDA) and anterior cervical discectomy and fusion (ACDF) in Cervical degenerative disc diseases. Methods: Two researchers independently conducted a search of PubMed, EMBASE, and Cochrane Central Register of Controlled Trails (CENTRAL) for randomized controlled trials (RCTs) following the Cochrane methodology guidelines. A fixed-effects or random-effects model was applied based on different heterogeneity. Review Manager (Version 5.4.1) software was used to perform data analysis. Results: A total of 8 RCT studies were included in this meta-analysis. The results indicate that the DCDA group had a higher incidence of reoperation (P = 0.03) and a lower incidence of ASD (P = 0.04) than the CDA group. There was no significant difference between two groups regarding NDI score (P = 0.36), VAS ARM score (P = 0.73), VAS NECK score (P = 0.63), EQ-5D score (P = 0.61) and dysphagia incidence (0.18). Conclusion: DCDA and ACDF have similar results in terms of NDI scores, VAS scores, EQ-5D scores, and dysphagia. In addition, DCDA can reduce the risk of ASD but increases the risk of reoperation.

Prognostic impact of colorectal cancer patients with bone metastases: a single-center experience.

The incidence of bone metastasis (BM) in colorectal cancer (CRC) patients is low and the prognosis is poor. There is no clear conclusion on the risk factors affecting the survival of CRC patients with BM. The aim of this study was to investigate the factors that may affect the prognosis of CRC patients with BM. The clinical and pathological data of CRC patients with BM were retrospectively analyzed. The overall survival after BM diagnosis was estimated using the Kaplan-Meier method and Log-rank test, and a multivariable cox regression model was used to identify the prognostic factors of overall survival. This study included 178 CRC patients with BM, of whom 151 had left-sided CRC and 27 had right-sided colon cancer. 1124 CRC patients with BM from the SEER database were included to perform a sensitivity analysis of the primary outcome. Multivariate analysis showed that the N staging, site of BM, and primary tumor sidedness (PTS) were independent prognostic factors for CRC with BM. Among them, right-sided colon cancer patients with BM had a poorer prognosis. Sensitivity analyses showed that PTS was an independent prognostic factor in CRC patients with BM. Primary tumor sidedness and N stage may be potential prognostic markers for BM of CRC. The prognosis of N0 stage CRC with BM is better, while the prognosis of right-sided colon cancer is poor.

Sequential Therapy for Bone Regeneration by Cerium Oxide-Reinforced 3D-Printed Bioactive Glass Scaffolds.

Rational design of multifunctional biomaterials with customized architecture and on demand bioactivity is of great significance for bone tissue engineering (BTE) in modern society. Herein, a versatile therapeutic platform has been established by integrating cerium oxide nanoparticles (CeO2 NPs) into bioactive glass (BG) to fabricate three-dimensional (3D)-printed scaffolds, achieving a sequential therapeutic effect against inflammation and promoting osteogenesis toward bone defect. The antioxidative activity of CeO2 NPs plays a crucial role in alleviating the oxidative stress upon formation of bone defects. Subsequently, CeO2 NPs exert a promotion effect on the proliferation and osteogenic differentiation of rat osteoblasts through enhancing mineral deposition and alkaline phosphatase and osteogenic gene expression. Strikingly, the incorporation of CeO2 NPs bestows on the BG scaffolds greatly reinforced mechanical properties, improved biocompatibility, adequate cell adhesion, elevated osteogenic capability, and multifunctional performance in a single platform. In vivo studies on the treatment of rat tibial defect confirmed the better osteogenic properties of CeO2-BG scaffolds compared with pure BG scaffolds. Additionally, the employment of the 3D printing technique creates a proper porous microenvironment around the bone defect, which further facilitates the cell in-growth and new bone formation. This report provides a systematic study on CeO2-BG 3D-printed scaffolds prepared by simple ball milling method, achieving sequential and integral treatment in BTE based on a single platform.

A Unified Therapeutic-Prophylactic Tissue-Engineering Scaffold Demonstrated to Prevent Tumor Recurrence and Overcoming Infection toward Bone Remodeling.

Osteosarcoma occurs in children and adolescents frequently and leads to a high fatality rate. Although surgical resection is the most common methods in clinic, patients always suffer from tumor metastasis and recurrence and it is difficult for them to self-repair large bone defects. Furthermore, the postoperative infection from bacteria triggers an inflammatory response and hinders the bone-repair process. This work demonstrates a gadolinium (Gd)-complex and molybdenum sulfide (MoS2 ) co-doped N-acryloyl glycinamide (NAGA)/gelatin methacrylate (Gel-MA) multifunctional hydrogel (GMNG). The combination between NAGA and Gel-MA endows the GMNG with attractive mechanical properties and controllable degradation ability. The MoS2 improves the hydrogel system, which has excellent photothermal ability to kill tumor cells and inhibit bacterial infection both in vitro and in vivo. Based on the Gd-complex, the magnetic resonance imaging (MRI) effect can be used to monitor the position and degradation situation of the hydrogel. Notably, accompanied by the degradation of GMNG hydrogel, the gradually released Gd3+ from the hydrogel exhibits osteogenic property and could promote new bone formation efficiently in vivo. Therefore, this strategy supplies a method to prepare multifunctional bone-defect-repair materials and is expected to represent a significant guidance and reference to the development of biomaterials for bone tissue engineering.

[The analysis of prognostic factors in patients with thoracic ossification of the ligamentum flavum and thoracic ossification of posterior longitudinal ligament by posterior decompression].

OBJECTIVE: To investigate the prognostic factors for patients with thoracic ossification of the ligamentum flavum (OLF) and thoracic ossification of posterior longitudinal ligament (OPLL). METHODS: Clinical information of 83 patients suffering from thoracic OLF and OPLL was reviewed retrospectively from January 2006 to June 2010. The related factors such as gender, age, preoperative and postoperative Japanese Orthopedic Association (JOA) score, pathological segment, type of thoracic OPLL, degree of thoracic kyphosis, anteroposterior diameter of OPLL, range of circumferential decompression, cerebrospinal fluid leakage or not and dysfunction or not and carotid lumbar disorders or not were analyzed by Chi-square and Logistic regression. RESULTS: All cases were classified into desirable group (58 cases) and undesirable group (25 cases) based on the postoperative JOA score improvement rate. Comparison of physical characteristics between two groups of age, preoperative JOA and the course of the disease had not statistically significant (P > 0.05). Two groups in pathological segment of thoracic OPLL (χ(2) = 6.290, P = 0.043), the ossification type of OPLL (χ(2) = 5.361, P = 0.021) and dysfunction or not in preoperative (χ(2) = 27.711, P = 0.000) had significant difference. Logistic regression analysis showed that the upper thoracic segments (P = 0.044), beak type ossification (P = 0.023) and with dysfunction in preoperative (P = 0.009) were risk factors. There were 24 patients (28.9%) with cerebrospinal fluid leakage, 3 patients with early postoperative deep infection and neurological deterioration of 2 cases in postoperative. CONCLUSIONS: Patients with ossification on the upper section of thoracic have a better prognosis, but the beaked localized longitudinal ligament ossification in patients and associated with preoperative dysfunction show a poor prognosis, combined jumping segmental ossification and cervical or lumbar severe disorders are the influencing factor for poor prognosis.

Causal effects of homocysteine levels on the components of sarcopenia: A two-sample mendelian randomization study.

Background: Currently, it is unclear whether there is a causal association between genetically predicted plasma homocysteine (Hcy) levels and the risk of sarcopenia. We performed a Mendelian randomization (MR) study to assess the association between circulating Hcy levels and the components [grip strength, walking pace, and appendicular lean mass (ALM)] of sarcopenia. Methods: Independent single nucleotide polymorphisms (SNPs) significantly associated with plasma Hcy levels served as instrumental variables. Summary-level data regarding the components of sarcopenia. Were obtained from the UK Biobank. Inverse variance weighted (IVW) as the primary method was used for Mendelian randomization (MR) analysis. We also use four models, weighted median, MR-Egger regression, Maximum likelihood, and Penalised weighted median, as supplementary methods to IVW. The MR-Egger intercept test, Cochran's Q test, and "leave-one-out" sensitivity analysis were performed to evaluate the horizontal pleiotropy, heterogeneities, and stability of the causal association between Hcy levels and the components of sarcopenia. Results: The IVW-MR analysis suggested significant negative associations of increased plasma Hcy levels with grip strength (right: effect = -0.036, SE = 0.032, p = 5.53E-4; left: effect = -0.045, SE = 0.010, p = 1.45E-5), walking pace (effect = -0.038, SE = 0.011, p = 3.18E-4), and ALM (effect = -0.058, 0.013, p = 1.03E-5). However, there were no significant associations of decreased plasma Hcy levels with grip strength (right: effect = 0.005, SE = 0.021, p = 0.82; left: effect = -0.006, SE = 0.014, p = 0.64), walking pace (effect = 0.01, 0.020, p = 0.61), or ALM (effect = -0.034, SE = 0.018, p = 0.06).The accuracy and robustness of these findings were confirmed by sensitivity tests. Conclusion: Increased circulating Hcy levels were associated with lower grip strength, slower walking pace, and decreased ALM.