Glaucocalyxin A delays the progression of OA by inhibiting NF-κB and MAPK signaling pathways.

BACKGROUND: Osteoarthritis (OA) is a common degenerative joint condition marked by inflammation and cartilage breakdown. Currently, there is a dearth of treatment medications that can clearly slow the course of OA. Glaucocalyxin A (GLA) is a diterpene chemical identified and extracted from Rabdosia japonica with antithrombotic, anticoagulant, anti-tumor, anti-inflammatory, anti-oxidant, and other pharmacological properties. Previous research has linked inflammation to abnormalities in the homeostasis of the extracellular matrix (ECM). Although GLA has been shown to have anti-inflammatory qualities, its effects on the progression of OA are unknown. As a result, the goal of this study was to see if GLA could slow the course of OA. METHODS: ATDC5 cells were stimulated by IL-1ß to create an inflammatory chondrocyte damage model. Quantitative polymerase chain reaction, Western Blot, high-density culture, and immunofluorescence were used to detect the expression levels of associated gene phenotypes. We also created a mouse model of OA induced by destabilization of the medial meniscus (DMM) instability, and GLA was administered intraperitoneally once every two days for eight weeks. Mice knee specimens were stained with hematoxylin-eosin, Safranin O/fast green, and immunohistochemical, and the Osteoarthritis Research Society International grade system and Mankin's score were used to assess the protective effect of GLA on cartilage. RESULTS: In vitro and in vivo, we explored the effects and molecular processes of GLA as a therapy for OA. The findings demonstrated that GLA might reduce the expression of associated inflammatory mediators and protect the ECM by inhibiting the NF-κB and MAPK signaling pathways. Animal research revealed that GLA could protect against the DMM-induced OA model mice by stabilizing ECM. CONCLUSION: Taken together, our findings show that GLA has a protective impact on cartilage throughout OA progression, implying that GLA could be employed as a possible therapeutic agent for OA, thus giving a new therapeutic method for the treatment of OA.

Mechanism of lysine oxidase-like 1 promoting synovial inflammation mediating rheumatoid arthritis development.

Rheumatoid arthritis (RA) is a chronic inflammatory joint disease that causes great distress to patients and society. Early diagnosis is the key to the successful treatment of RA. The basement membrane, one of the oldest tissue structures, is localized under the epithelium. Its complex composition and rich biological functions have made it a focus of research in recent years, while basement membrane-associated genetic variants are involved in most human disease processes. The aim of this study is to find new diagnostic biomarkers for RA and explore their role and possible mechanism in rheumatoid arthritis. The GSE12021, GSE55235 and GSE55457 datasets were downloaded from the GEO database. Their fraction associated with basement membrane genes was analyzed and differentially expressed genes between the disease and normal groups were explored. We identified two basement membrane-associated genes, lysine oxidase-like 1 (LOXL1) and discoid peptide receptor 2 (DDR2). Focusing on the more interesting LOXL1, we found that LOXL1 expression was significantly elevated in the synovium of patients with rheumatoid arthritis, and LOXL1 mRNA and protein levels were elevated in tumor necrosis factor α-stimulated human synovial sarcoma cells (SW982). And LOXL1 knockdown inhibited tumor necrosis factor α-induced inhibition in SW982 cells expression of inducible nitric oxide synthase (INOS), cyclooxygenase-2 (COX2), and interleukin-6 (IL-6). Interestingly, knockdown of LOXL1 inhibited the phosphorylation of PI3K and AKT. In summary, LOXL1 may become a novel diagnostic gene for RA, and knockdown of LoxL1 may inhibit synovial inflammation by affecting PI3K/AKT pathway.

A web-based calculator for predicting the prognosis of patients with sarcoma on the basis of antioxidant gene signatures.

BACKGROUND: Oxidative stress plays a critical role in tumorigenesis, tumor development, and resistance to therapy. A systematic analysis of the interactions between antioxidant gene expression and the prognosis of patients with sarcoma is lacking but urgently needed. METHODS: Gene expression and clinical data of patients with sarcoma were derived from The Cancer Genome Atlas Sarcoma (training cohort) and Gene Expression Omnibus (validation cohorts) databases. Least absolute shrinkage, selection operator regression, and Cox regression were used to develop prognostic signatures for overall survival (OS) and disease-free survival (DFS). Based on the signatures and clinical features, two nomograms for predicting 2-, 4-, and 6-year OS and DFS were established. RESULTS: On the basis of the training cohort, we identified five-gene (CHAC2, GPX5, GSTK1, PXDN, and S100A9) and six-gene (GGTLC2, GLO1, GPX7, GSTK1, GSTM5, and IPCEF1) signatures for predicting OS and DFS, respectively, in patients with sarcoma. Kaplan-Meier survival analysis of the training and validation cohorts revealed that patients in the high-risk group had a significantly poorer prognosis than those in the low-risk group. On the basis of the signatures and other independent risk factors, we established two models for predicting OS and DFS that showed excellent calibration and discrimination. For the convenience of clinical application, we built web-based calculators (OS: https://quankun.shinyapps.io/sarcDFS/). CONCLUSIONS: The antioxidant gene signature models established in this study can be novel prognostic predictors for sarcoma.

Analysis of Risk Factors for Non-union After Surgery for Limb Fractures: A Case-Control Study of 669 Subjects.

Objective: The purpose of this study was to analyze the risk factors for limb fracture non-union in order to improve non-union prevention and early detection. Methods: A total of 223 patients with non-union after surgery for limb fractures performed at our institution from January 2005 to June 2017 were included as the case group, while a computer-generated random list was created to select 446 patients with successful bone healing after surgery for limb fractures who were treated during the same period as the control group, thus achieving a ratio of 1:2. The medical records of these patients were reviewed retrospectively. Age, sex, body mass index, obesity, smoking, alcohol, diabetes, hypertension, osteoporosis, fracture type, multiple fractures, non-steroidal anti-inflammatory drugs (NSAIDs) use, delayed weight bearing, internal fixation failure, and infection data were analyzed and compared between the two groups. A multivariate logistic regression model was constructed to determine relevant factors associated with non-union. Results: After comparison between two groups by univariate analysis and multivariate logistic regression, we found some risk factors associated that osteoporosis (odds ratio [OR] = 3.16, 95% confidence interval [CI]: 2.05-4.89, p < 0.001), open fracture (OR = 2.71, 95%CI: 1.72-4.27, p < 0.001), NSAIDs use (OR = 2.04, 95%CI: 1.24-3.37, p = 0.005), delayed weight bearing (OR = 1.72, 95%CI: 1.08-2.74, p = 0.023), failed internal fixation (OR = 5.93, 95%CI: 2.85-12.36, p < 0.001), and infection (OR = 6.77, 95%CI: 2.92-15.69, p < 0.001) were independent risk factors for non-union after surgery for limb fractures. Conclusions: Osteoporosis, open fracture type, NSAIDs use, delayed weight bearing, failed internal fixation, and infection were found to be the main causes of bone non-union; clinicians should, therefore, take targeted measures to intervene in high-risk groups early.

A SEER-based nomogram accurately predicts prognosis in Ewing's sarcoma.

Ewing's sarcoma is a high-grade malignancy bone and soft tissue tumor that most commonly occurs in children and adolescents. Although the overall prognosis of Ewing's sarcoma has improved, the 5-year survival rate has not improved significantly. The study aimed to determine the risk factors independently associated with the prognosis of Ewing's sarcoma and to construct a nomogram to predict patient survival. Patients diagnosed with Ewing's sarcoma were collected from the Surveillance, Epidemiology, and End Results program database between 2004 and 2015 and further divided into training and validation cohort. Univariate and multivariate Cox regression analyses were used to identify meaningful independent prognostic factors. The nomogram was used to predict 3- and 5-year overall survival (OS) and cancer-specific survival (CSS). Finally, the nomogram was verified internally and externally through the training and validation cohorts, and the predictive capability was evaluated using the receiver operating characteristic (ROC) curve, C-index, and calibration curve and compared with that of the 7th TNM stage. A total of 1120 patients were divided into training (n = 713) and validation (n = 407) cohorts. Based on the multivariate analysis of the training cohort, a nomogram that integrated age, tumor size, primary site, N stage, and M stage was constructed (P < 0.05). The predicted C-indexes of OS and CSS of the training cohort were 0.744 (95% CI 0.717-0.771) and 0.743 (95% CI 0.715-0.770), respectively. However, the TNM stage had a C-index of 0.695 (95% CI 0.666-0.724) and 0.698 (95% CI 0.669-0.727) for predicting OS and CSS, respectively. The nomogram showed higher C-indexes than those in the TNM stage. Furthermore, the internal and external calibration curves showed good consistency between the predicted and observed values. Age, tumor size, primary site, N stage, and M stage are independent risk factors affecting the OS and CSS in Ewing's sarcoma patients. Compared with the 7th TNM staging, the nomogram consisting of these factors was more accurate for risk assessment and survival prediction in patients with Ewing's sarcoma, thus providing a novel reliable tool for risk assessment and survival prediction in Ewing's sarcoma patients.

Celastrol Attenuates RANKL-Induced Osteoclastogenesis in vitro and Reduces Titanium Particle-Induced Osteolysis and Ovariectomy-Induced Bone Loss in vivo.

Excessive bone resorption by osteoclasts contributes significantly to osteoclast-related diseases such as periprosthetic osteolysis and osteoporosis. Osteolysis in a titanium particle-induced calvarial model and bone loss in an ovariectomized mice model occurred similarly to those in humans; thus, these models can be used to evaluate potential therapies for aseptic prosthetic loosening and osteoporosis. Celastrol, which is extracted from the seeds of the genus Tripterygium, has been thoroughly investigated for its anti-inflammatory and anti-cancer pharmacological effects. However, the mechanisms involving bone metabolism by which celastrol inhibits osteoclastogenesis are not yet fully understood. We demonstrated that celastrol inhibited the receptor activator of nuclear factor κB ligand-induced osteoclastogenesis and the bone resorptive function of osteoclasts in vitro by inhibiting the activation of transforming growth factor ß-activated kinase 1-mediated NF-κB and mitogen-activated protein kinase signaling pathways and downregulating osteoclastogenesis marker-related genes. Furthermore, celastrol was also shown to be beneficial in both the titanium particle-induced osteolysis calvarial and the murine ovariectomy-induced bone loss. Collectively, our results suggested that celastrol is promising for the prevention of aseptic prosthetic loosening and osteoporosis in the treatment of osteolytic diseases induced by disrupted osteoclast formation and function.

An Intraoperative Trajectory-Determined Strategy of Patient-Specific Drill Template for C2 Transoral Pedicle Insertion in Incomplete Reduction of Atlantoaxial Dislocation: An In Vitro Study.

OBJECTIVES: This study aims to explore a novel intraoperative trajectory-determined strategy of grouped patient-specific drill templates (PDTs) for transoral C2 pedicle screw insertion (C2 TOPI) for atlantoaxial dislocation (AAD) with incomplete reduction and to evaluate its efficiency and accuracy. METHODS: Ten cadaveric C2 specimens were scanned by computed tomography (CT) and randomly divided into two groups (the PDT and freehand groups). A novel intraoperative trajectory-determined strategy of grouped PDTs was created for AAD with incomplete reduction. C2 TOPI was performed by use of the PDT technique and the fluoroscopy-guided freehand technique. After surgery, the screw deviations from the centroid of the cross-section at the midpoint of the pedicle and screw position grades were assessed in both groups. RESULTS: Compared to the freehand group, the PDT group had a significantly shorter surgery time than the freehand group (47.7 vs 61.9 min, P < 0.001). The absolute deviations from the centroids between the preoperative designs and postoperative measurements on the axial plane of the pedicle were 1.19 ± 0.25 mm in the PDT group and 1.82 ± 0.51 mm in the freehand group. On the sagittal plane of the pedicle, the corresponding values were 1.10 ± 0.33 mm in the PDT group and 1.70 ± 0.49 mm in the freehand group. The absolute deviations of the free-hand group on both the axial and sagittal planes were higher than that of the freehand group (P < 0.05 and P < 0.05, respectively). For the grade of screw insertion position, nine (90%) were observed in type I and one (10%) in type II in the PDT group, whereas five (50%) were in type I, three (30%) were in type II, and two (20%) in type III in the freehand group. Statistical differences could not be found between the groups in terms of the screw positions (P > 0.05). CONCLUSION: The novel intraoperative trajectory-determined strategy of grouped PDTs can be used as an accurate and feasible method for C2 TOPI for AAD with incomplete reduction.

Glaucocalyxin A suppresses osteoclastogenesis induced by RANKL and osteoporosis induced by ovariectomy by inhibiting the NF-κB and Akt pathways.

ETHNOPHARMACOLOGICAL RELEVANCE: Glaucocalyxin A (GLA), the most abundant active component of the aboveground sections of Rabdosia japonica (Burm. f.) Hara var. glaucocalyx (Maxim.) Hara, possesses various pharmacological activities, such as antioxidant, antithrombosis, anticoagulation, antibacterial, antitumor, anti-inflammatory activities. According to previous studies, inflammation is closely associated with osteoclast differentiation and activity. Although GLA has demonstrated effective anti-inflammatory properties, its effects on osteoclast differentiation remain unclear. AIM OF THE STUDY: To examine the possible inhibitory effects of GLA and its molecular mechanisms in osteogenesis induced by RANKL as well as ovariectomy (OVX)-induced osteoporosis (OP) in mice. MATERIALS AND METHODS: Tartrate-resistant acid phosphatase (TRAP) staining, F-actin staining, and a bone resorption pit assay were applied for identifying the effects of GLA on the differentiation of osteoclasts and the function of bone resorption. The mRNA expression of the genes related to osteoclast differentiation was measured by quantitative PCR. Protein expression of nuclear factor of activated T-cells, cytoplasmic 1 (NFATc1), c-fos and phosphorylation of inhibitor of nuclear factor kappa B (IκBα), protein kinase B (AKT), c-Jun N-terminal kinase (JNK), extracellular signal-regulated kinase (ERK), and p38 in RANKL-induced osteoclasts was determined using western blotting. The effect of GLA on OP was studied using a mouse model of OVX. RESULTS: At nontoxic concentrations ≤0.5 µM in vitro, GLA suppressed the formation of osteoclasts induced by RANKL with the decreased number and area size of TRAP-positive multinuclear osteoclasts, and the resorption of bone function by reducing F-actin ring number and bone resorption pit areas. It also reduced the expression of the genes specific for osteoclasts, which included genes encoding NFATc1, cathepsin K, c-fos, TRAP, vacuolar-type ATPase d2, and dendritic cell-specific transmembrane protein. Moreover, GLA repressed NF-κB and Akt pathway activation induced by RANKL. Micro-CT analysis of femur samples indicated decreased bone loss and greater trabecular bone density after GLA treatment, which showed that GLA played a protective role by inhibiting bone loss in OVX-induced OP mice in vivo. CONCLUSIONS: Our study is the first to show that GLA has significant therapeutic potential in OP, which is the disease of osteoclast increase caused by estrogen deficiency.

Patient-specific drill template for C2 transoral pedicle insertion in complete reduction of atlantoaxial dislocation: cadaveric efficacy and accuracy assessments.

BACKGROUND: The transoral atlantoaxial reduction plate (TARP) is an effective advance in the treatment of atlantoaxial dislocation (AAD) and can enable the performance of anterior atlantoaxial release, reduction, decompression, and internal fixation in a one-stage operation. However, accurate transoral C2 pedicle insertion (C2TOPI) remains a challenge. The aim of this study is to develop a grouped patient-specific drill template (PDT) specifically for AAD with complete reduction and, furthermore, to compare its efficacy and accuracy in facilitating C2TOPI. METHODS: After CT scanning, ten cadaveric C2 specimens were randomly assigned to two groups (the PDT and freehand group). A grouped PDT specifically for AAD with complete reduction was designed and manufactured. C2TOPI was performed using the PDT or the fluoroscopy-guided freehand technique. Postoperative CT scans were subsequently performed to analyze the deviations at the centroid of the cross section at the midpoint of the pedicle. Screw position grades were also assessed in both groups. RESULTS: Compared to the freehand group, the PDT group had a significantly shorter surgery time (p < 0.001). Significant differences between the two groups were observed in the absolute value of the deviations at the centroid of the pedicle on either the axial or sagittal planes (p < 0.05). No significant difference was found in the screw positions between the two groups (p > 0.05); however, two unacceptable breaches (20%) occurred in the freehand group. CONCLUSION: A specifically designed PDT could provide an accurate and easy-to-apply method for C2TOPI in AAD with complete reduction.

Is a patient-specific drill template via a cortical bone trajectory safe in cervical anterior transpedicular insertion?

BACKGROUND: This study aimed to develop patient-specific drill templates by computer numerical control or three-dimensional printing via two cortical bone trajectories (CBTs) and to evaluate their efficacies and accuracies in cervical anterior transpedicular insertion. METHODS: Preoperative CT images of 20 cadaveric cervical vertebrae (C3-C7) were obtained. After image processing, patient-specific drill templates were randomly assigned to be constructed via two CBTs (CBT0 and CBT0.7) and manufactured by two methods (computer numerical control and three-dimensional printing). Guided by patient-specific drill templates, 3.5-mm-diameter screws were inserted into the pedicles. Postoperative CT scans were performed to evaluate the screw deviation in the entry point and midpoint of the pedicle. The screw positions were also graded. RESULTS: Computer numerical control patient-specific drill templates had a significantly shorter manufacturing time compared to three-dimensional-printed patient-specific drill templates (p < 0.01). Absolute deviations at the entry point and midpoint of the pedicle had no significant differences on the transverse and sagittal planes (p > 0.05). There were no significant differences in screw positions (p = 0.3). However, three screw positions were in grade 3 in CBT0, while the others were in grade 1. CONCLUSIONS: CBT0.7 appears to be a safe and feasible trajectory for cervical anterior transpedicular insertion. Bio-safe computer numerical control patient-specific drill templates can facilitate cervical anterior transpedicular insertion with good feasibility and accuracy.

Tibial tunnel placement in anatomic anterior cruciate ligament reconstruction: a comparison study of outcomes between patient-specific drill template versus conventional arthroscopic techniques.

INTRODUCTION: Accurate anatomic graft tunnel positioning is essential for the successful application of anatomic anterior cruciate ligament (ACL) reconstruction. The accurate insertion of the tibial tunnel (TT) remains challenging. Here, we explored a novel strategy of patient-specific drill template (PDT) for the placement of TT in ACL reconstruction and assessed its efficacy and accuracy. MATERIALS AND METHODS: TT placement was randomized and performed by use of the PDT technique in 40 patients (PDT group) and the conventional arthroscopic technique in 38 patients (Arthroscopic group). After surgery, the deviations at the center point of the ACL tibial attachment area and radiological TT positioning were assessed in both groups. The preoperative and follow-up examinations included pivot-shift testing, KT-1000 arthrometer testing, the Lysholm and International Knee Documentation Committee scales were used to compare the knee stability and the functional state. RESULTS: The ideal center points achieved in the PDT group were more precise than that in the arthroscopic group (p < 0.001). Radiological TT positioning performed by use of the PDT technique was more accurate than that by the arthroscopic technique (p = 0.027). Statistical differences could not be found between the groups in terms of the pivot-shift test, KT-1000 arthrometer laxity measurements, the Lysholm or International Knee Documentation Committee scales. Both groups improved at follow-up compared with the preoperative assessment in terms of the pivot-shift test, the laxity tests, and scoring scales. CONCLUSIONS: The novel PDT strategy could provide more accurate TT positioning than the traditional arthroscopic technique in ACL reconstruction. However, functional scales and stability tests gave similar results in the PDT and the standard techniques. LEVEL OF EVIDENCE: I.