Therapeutic impacts of GNE­477­loaded H2O2 stimulus­responsive dodecanoic acid­phenylborate ester­dextran polymeric micelles on osteosarcoma.

Osteosarcoma (OS) is a highly malignant primary bone neoplasm that is the leading cause of cancer­associated death in young people. GNE­477 belongs to the second generation of mTOR inhibitors and possesses promising potential in the treatment of OS but dose tolerance and drug toxicity limit its development and utilization. The present study aimed to prepare a novel H2O2 stimulus­responsive dodecanoic acid (DA)­phenylborate ester­dextran (DA­B­DEX) polymeric micelle delivery system for GNE­477 and evaluate its efficacy. The polymer micelles were characterized by morphology, size and critical micelle concentration. The GNE­477 loaded DA­B­DEX (GNE­477@DBD) tumor­targeting drug delivery system was established and the release of GNE­477 was measured. The cellular uptake of GNE­477@DBD by three OS cell lines (MG­63, U2OS and 143B cells) was analyzed utilizing a fluorescent tracer technique. The hydroxylated DA­B was successfully grafted onto dextran at a grafting rate of 3%, suitable for forming amphiphilic micelles. Following exposure to H2O2, the DA­B­DEX micelles ruptured and released the drug rapidly, leading to increased uptake of GNE­477@DBD by cells with sustained release of GNE­477. The in vitro experiments, including MTT assay, flow cytometry, western blotting and RT­qPCR, demonstrated that GNE­477@DBD inhibited tumor cell viability, arrested cell cycle in G1 phase, induced apoptosis and blocked the PI3K/Akt/mTOR cascade response. In vivo, through the observation of mice tumor growth and the results of H&E staining, the GNE­477@DBD group exhibited more positive therapeutic outcomes than the free drug group with almost no adverse effects on other organs. In conclusion, H2O2­responsive DA­B­DEX presents a promising delivery system for hydrophobic anti­tumor drugs for OS therapy.

Comprehensive AI-assisted tool for ankylosing spondylitis based on multicenter research outperforms human experts.

Introduction: The diagnosis and treatment of ankylosing spondylitis (AS) is a difficult task, especially in less developed countries without access to experts. To address this issue, a comprehensive artificial intelligence (AI) tool was created to help diagnose and predict the course of AS. Methods: In this retrospective study, a dataset of 5389 pelvic radiographs (PXRs) from patients treated at a single medical center between March 2014 and April 2022 was used to create an ensemble deep learning (DL) model for diagnosing AS. The model was then tested on an additional 583 images from three other medical centers, and its performance was evaluated using the area under the receiver operating characteristic curve analysis, accuracy, precision, recall, and F1 scores. Furthermore, clinical prediction models for identifying high-risk patients and triaging patients were developed and validated using clinical data from 356 patients. Results: The ensemble DL model demonstrated impressive performance in a multicenter external test set, with precision, recall, and area under the receiver operating characteristic curve values of 0.90, 0.89, and 0.96, respectively. This performance surpassed that of human experts, and the model also significantly improved the experts' diagnostic accuracy. Furthermore, the model's diagnosis results based on smartphone-captured images were comparable to those of human experts. Additionally, a clinical prediction model was established that accurately categorizes patients with AS into high-and low-risk groups with distinct clinical trajectories. This provides a strong foundation for individualized care. Discussion: In this study, an exceptionally comprehensive AI tool was developed for the diagnosis and management of AS in complex clinical scenarios, especially in underdeveloped or rural areas that lack access to experts. This tool is highly beneficial in providing an efficient and effective system of diagnosis and management.

Circ_0001060 Upregulates and Encourages Progression in Osteosarcoma.

Circular RNA (circRNA) is involved in the occurrence and development of various cancers. To this day, the expression and mechanism of circRNA in osteosarcoma (OS) remain unclear. We previously found that circ_0001060 was highly expressed in OS tumor tissues. In this work, we identified that high level expression of circ_0001060 was significantly associated with late clinical stage, larger tumor volume, higher frequency of metastasis, and poor prognosis in OS patients. Furthermore, we confirmed that silencing circ_0001060 inhibited the proliferation and migration of OS cell. Using bioinformatics analysis, we built three circRNA-miRNA-mRNA regulatory modules (circ_0001060-miR-203a-5p-TRIM21, circ_0001060-miR-208b-5p-MAP3K5, and circ_0001060-miR-203a-5p-PRKX), suggesting that these signaling axes may be involved in the inhibitory effect of circ_0001060 on OS. To sum up, circ_0001060 is a novel tumor biomarker for OS as well as a potential therapeutic target.

Isoliensinine Suppresses Osteoclast Formation Through NF-κB Signaling Pathways and Relieves Ovariectomy-Induced Bone Loss.

Osteoporosis is among the major contributors of pathologic fracture in postmenopausal women, which is caused by the bone metabolic disorder owing to the over-activation of osteoclasts. Inhibition of osteoclast differentiation and maturation has become a mainstream research interest in the prevention of osteoporosis. Isoliensinine (Iso) is a dibenzyl isoquinoline alkaloid with antioxidant, anti-inflammatory, and anti-cancer activities. However, whether it can be used as a potential treatment for osteoporosis remains undiscovered. Here, we investigated whether Iso might suppress the differentiation of osteoclasts in vitro and in vivo to play an anti-osteoporosis role. Our results showed that Iso inhibits the formation of mature multinuclear osteoclasts induced by RANKL, the bone resorption, and the osteoclast-specific genes expression by blocking the nuclear translocation of NF-κB p65, and the effect was in a dosage-dependent way. Furthermore, we investigated the therapeutic effect of Iso on osteoporosis in ovariectomized (OVX) mice. We found that Iso attenuated bone loss in the OVX mice and significantly promoted BS, Conn. DN, Tb.Th, TB.N, and BV/TV Index. All in all, Iso showed a prominent effect of osteoclast inhibition, with great promise for treating osteoporosis.

Diagnosis and treatment of primary hyperparathyroidism with pathological fracture of the limbs: A retrospective observational study.

Primary hyperparathyroidism (PHPT) with pathological fracture is rare, and the early symptoms of PHPT lack specificity, leading to misdiagnosis. Therefore, this study aimed to summarize the clinical characteristics and treatment of PHPT patients with pathological fractures and to improve the attention of orthopedic clinicians to PHPT. It is a retrospective study, 2226 patients with hyperparathyroidism in our hospital from 2009 to 2019 were screened, excluding secondary hyperparathyroidism and patients without limb fracture, and the remaining 20 patients with PHPT accompanied by pathological fractures were finally analyzed. Parathyroid hormone (PTH) and calcium levels were compared on the first postoperative day, and the prognosis of the patients was assessed by bone mineral density and Visual Analogue Scale scores at 3 and 12 months postoperatively. The early symptoms of PHPT patients in this study included urinary calculi (80%), bone pain (30%), and digestive tract symptoms (25%). Fourteen (70%) cases were misdiagnosed at the initial diagnosis. After parathyroidectomy, the blood calcium and PTH levels decreased significantly in all patients (P < .05). For the treatment of fracture, 9 of the patients underwent surgical treatment of the fracture, while the remaining patients received splint external fixation. The follow-up time was 4.60 ± 0.62 years (1-10 years). All patients recovered well from the fracture, the symptoms of systemic bone pain were markedly improved, and bone mineral density was significantly improved after surgery. Orthopedic surgeons need to avoid misdiagnosis and pay attention to the early symptoms in PHPT patients with pathological fracture, and better therapeutic effects can be obtained by combining parathyroidectomy with fractures fixation.

Aneurysmal bone cyst secondary to giant cell tumor of the extremities: a case series of 30 patients.

The purpose was to investigate the clinical features, diagnosis, treatment, and prognosis of aneurysmal bone cyst (ABC) secondary to giant cell tumors (GCT) of the extremities. Data from patients with ABC secondary to GCT of the extremities were obtained from the medical records. Clinical features, imaging findings, pathologic diagnosis, surgical methods, and prognosis were analyzed. The median age of the patients was 33 years (range 15 to 52 years) and 83.3 percent were between 20 to 40 years. The lesions were mainly located in the proximal tibia and distal femur, accounting for 63.3% (19/30). 21 patients were treated with curettage, and 9 with tumor resection. The recurrence rates of the curettage group and resection group were 52.4% and 11.1% respectively. However, the average postoperative (Musculoskeletal Tumor Society) MSTS score were 28.6±1.2 post-curettage, and 25.0±0.5 post-resection, with a significant difference between the 2 groups (P<0.01). In these relapsed patients, 10 underwent a second curettage, while 2 cases underwent a resection and there was no postoperative re-recurrence in both groups. A comprehensive analysis should be performed when making the diagnosis of ABC secondary to GCT. Although the recurrence rate is higher, curettage is still the optimal method for satisfactory joint function. If recurrence occurs after the first curettage, a second curettage should be performed.

ATG16L1 is a Potential Prognostic Biomarker and Immune Signature for Osteosarcoma: A Study Based on Bulk RNA and Single-Cell RNA-Sequencing.

BACKGROUND: Osteosarcoma is a common solid malignancy of the bone in children and adolescents, and its metastasis and recurrence are the principal causes of poor treatment outcomes. METHODS: Autophagy-related genes were used to cluster osteosarcoma patients by consensus clustering analysis using the GSE21257 database. Differentially expressed genes (DEGs) were identified by limma package. Multiple-gene risk signature was constructed using least absolute shrinkage and selection operator (LASSO) analysis and Cox regression analyses. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to determine gene expression levels. Then, single-cell RNA-sequencing dataset GSE152048 were used to identify the correlation between the DEGs and effector molecules expressed in specific tumor-infiltrating immune cells. RESULTS: Two clusters were identified in the consensus clustering analysis, which were confirmed by principal component analysis. Limma analysis revealed that 15 genes were related, and 9 genes were screened using protein-protein interaction network and LASSO regression analysis. Cox regression analyses identified 5 genes. Combined with survival analysis, only the autophagy related 16 like 1 gene (ATG16L1) was significant. The results of qRT-PCR showed low expression levels of ATG16L1 in tumor cells group. Immune infiltration analysis revealed significantly lower expression of CD8+ T cells in the high ATG16L1 gene expression group. ScRNA-seq revealed that in the ATG16L1+ CD8 + T cell group, the expression of GZMB was lower, whereas the expression of ITGA1 was higher. These results showed that ATG16L1 is an immune-related gene, which is associated with poor prognosis in patients with osteosarcoma. CONCLUSION: ATG16L1 is a potential prognostic biomarker and immune signature and may be a therapeutic target for osteosarcoma.

Single-Cell RNA Sequencing Reveals the Migration of Osteoclasts in Giant Cell Tumor of Bone.

Giant cell tumor of bone (GCTB) is benign tumor that can cause significant osteolysis and bone destruction at the epiphysis of long bones. Osteoclasts are thought to be highly associated with osteolysis in GCTB. However, the migration of osteoclasts in GCTB remains unclear. A deeper understanding of the complex tumor microenvironment is required in order to delineate the migration of osteoclasts in GCTB. In this study, samples were isolated from one patient diagnosed with GCTB. Single-cell RNA sequencing (scRNA-seq) was used to detect the heterogeneity of GCTB. Multiplex immunofluorescence staining was used to evaluate the cell subtypes identified by scRNA-seq. A total of 8,033 cells were obtained from one patient diagnosed with GCTB, which were divided into eight major cell types as depicted by a single-cell transcriptional map. The osteoclasts were divided into three subsets, and their differentiation trajectory and migration status were further analyzed. Osteoclast migration may be regulated via a series of genes associated with cell migration. Furthermore, four signaling pathways (RANKL, PARs, CD137 and SMEA3 signaling pathway) were found to be highly associated with osteoclast migration. This comprehensive single-cell transcriptome analysis of GCTB identified a series of genes associated with cell migration as well as four major signaling pathways that were highly related to the migration of osteoclasts in GCTB. Our findings broaden the understanding of GCTB bionetworks and provides a theoretical basis for anti-osteolysis therapy against GCTB in the future.

Moracin attenuates LPS-induced inflammation in nucleus pulposus cells via Nrf2/HO-1 and NF-κB/TGF-ß pathway.

The present study was designed to investigate the protective effect of moracin on primary culture of nucleus pulposus cells in intervertebral disc and explore the underlying mechanism. Moracin treatment significantly inhibited the LPS-induced inflammatory cytokine accumulation (IL-1ß, IL-6 and TNF-α) in nucleus pulposus cells. And moracin also dramatically decreased MDA activity, and increased the levels of SOD and CAT induced by LPS challenge. Moreover, the expressions of Nrf-2 and HO-1 were decreased and the protein levels of p-NF-κBp65, p-IκBα, p-smad-3 and TGF-ß were increased by LPS challenge, which were significantly reversed after moracin treatments. Moracin treatments also decreased the levels of matrix degradation enzymes (MMP-3, MMP-13) as indicated by RT-PCR analysis. However, Nrf-2 knockdown abolished these protective effects of moracin. Together, our results demonstrated the ability of moracin to antagonize LPS-mediated inflammation in primary culture of nucleus pulposus in intervertebral disc by partly regulating the Nrf2/HO-1 and NF-κB/TGF-ß pathway in nucleus pulposus cells.

Biomechanical Evaluation of an Anterior Upper Thoracic Plate Fixation System: An In Vitro Human Cadaveric Study.

OBJECTIVE: This study aimed to evaluate the biomechanical characteristics of the anterior upper thoracic plate fixation system (AUTP). METHODS: Twelve specimens were divided into 2 groups: the AUTP group and the anterior cervical locking plate (ACLP) group. Bone mineral density was assessed in all specimens. The specimens were loaded with pure bending moments of ±4 Nm to move toward extension/flexion, right/left lateral tilt, and right/left axial rotation. Each specimen was tested in 3 load cycles under 3 conditions (native, destabilized, and torsion). The tightening moments were assessed in every screw using a torsion meter. RESULTS: The tightening moment of the AUTP screws was more pronounced than the ACLP (P < 0.05). Significant decreases after stabilization with the AUTP compared with the native situation were observed in the AUTP group (P < 0.05). In terms of the direction of lateral tilt and torsion, the neutral zone increased significantly after stabilization with the AUTP compared with the native situation (P < 0.05). CONCLUSIONS: The AUTP was shown to provide more rigidity to the destabilized spine than the ACLP.

Comparisons of front plate, percutaneous sacroiliac screws, and sacroiliac anterior papilionaceous plate in fixation of unstable pelvic fractures.

BACKGROUND: This observational study was aimed at comparing the clinical efficacy of sacroiliac anterior plate fixation (SAPF), sacroiliac anterior papilionaceous plate (SAPP), and percutaneous sacroiliac screw internal fixation (PSCIF) introduced for patients with unstable pelvic fracture. METHODS: Seventy-eight patients with unstable pelvic fracture (Tile type B or C) were recruited. Twenty-six patients underwent SAPF, 26 underwent SAPP, and 26 underwent PSCIF. Matta scores were calculated to evaluate the reduction of pelvic fractures, and Majeed scores were applied for the assessment of functional recoveries after surgery. Other perioperative clinical indicators were also recorded, including operation time, bleeding status, length of incision, ambulation time, fracture healing time, and incision infection. RESULTS: Total operation time of PSCIF was remarkably shorter than that of SAPF and SAPP (P < .05), and the bleeding volume of SAPF and SAPP group was almost 26∼29 times as high as that of PSCIF group (P < .05). Besides, SAPP resulted in significant blood loss compared with SAPF (P < .05), while SAPF resulted in significantly larger operative incision length than SAPP and PSCIF (P < .05). Moreover, patients' stay time was prolonged in both SAPF and SAPP groups than in the PSCIF group (P < .05). Patients who received PSCIF exhibited significantly higher Matta and Majeed scores than those who received SAPF (all P < .05). Finally, SAPP was associated with fewer complications than SAPF, and complications were barely observed in the PSCIF group. CONCLUSION: PSCIF may be more appropriate for patients with unstable pelvic fractures in comparison with SAPP and SAPF. Besides, SAPP is likely to be more efficacious than SAPF especially for Tile C patients.

MicroRNA-17 family as novel biomarkers for cancer diagnosis: a meta-analysis based on 19 articles.

Cancer remains as the leading cause of death all over the world due to the lack of efficient diagnostic techniques and therapeutic methods. Many studies have reported the potential diagnostic value of microRNA-17 (miRNA-17, miR-17) family members as biomarkers for cancer detection. However, inconsistent results were revealed from a wide range of studies. As a result of this, a meta-analysis based on 19 studies was conducted to assess the diagnostic performance of miR-17 family for cancer detection. A total of 1772 patients with certain types of cancer and 1320 healthy controls were involved in these studies. The overall diagnostic accuracy was measured by the following: sensitivity, 0.67 (95 % confidence interval (CI) 0.60-0.74); specificity, 0.83 (95 % CI 0.74-0.85); positive likelihood ratio (PLR), 3.9 (95 % CI 2.6-5.9); negative likelihood ratio (NLR), 0.40 (95 % CI 0.34-0.48); and diagnostic odds ratio (DOR), 10 (95 % CI 6-16), respectively. Additionally, the pooled area under the summary receiver operator characteristic (SROC) curve (area under the curve (AUC)) was 0.79 (95 % CI 0.75-0.82), indicating a relatively low accuracy of miR-17 family as biomarkers for cancer detection. Subgroup analysis further showed that miR-17 family had more reliable performance in cancer diagnosis for Asian than that for Caucasian. Moreover, multiple miRNAs containing miR-17, -20a/b, and -93 reflected higher diagnostic accuracy than both miR-106a/b (single miRNA) and the overall miR-17 family assay. Therefore, appropriate combinations of miR-17 family may be used as non-invasive screening biomarkers for cancer, and it is necessary to carry out a large-scale population-based study to further assess the potential diagnostic value of miR-17 family.

Genetic diversity analysis reveals that geographical environment plays a more important role than rice cultivar in Villosiclava virens population selection.

Rice false smut caused by Villosiclava virens is an economically important disease of grains worldwide. The genetic diversity of 153 isolates from six fields located in Wuhan (WH), Yichang Wangjia (YCW), Yichang Yaohe (YCY), Huanggang (HG), Yangxin (YX), and Jingzhou (JZ) in Hubei province of China were phylogenetically analyzed to evaluate the influence of environments and rice cultivars on the V. virens populations. Isolates (43) from Wuhan were from two rice cultivars, Wanxian 98 and Huajing 952, while most of the other isolates from fields YCW, YCY, HG, YX, and JZ originated from different rice cultivars with different genetic backgrounds. Genetic diversity of isolates was analyzed using random amplified polymorphic DNA (RAPD) and single-nucleotide polymorphisms (SNP). The isolates from the same cultivars in Wuhan tended to group together, indicating that the cultivars had an important impact on the fungal population. The 110 isolates from individual fields tended to cluster according to geographical origin. The values of Nei's gene diversity (H) and Shannon's information index (I) showed that the genetic diversity among isolates was higher between than within geographical populations. Furthermore, mean genetic distance between groups (0.006) was higher than mean genetic distance within groups (0.0048) according to MEGA 5.2. The pairwise population fixation index (FST) values also showed significant genetic differentiation between most populations. Higher genetic similarity of isolates from individual fields but different rice cultivars suggested that the geographical factor played a more important role in the selection of V. virens isolates than rice cultivars. This information could be used to improve the management strategy for rice false smut by adjusting the cultivation measures, such as controlling fertilizer, water, and planting density, in the rice field to change the microenvironment.

[Establishment of a mouse model bearing spinal metastatic human prostate cancer using fluorescence-labeled PC-3 cells].

OBJECTIVE: To establish a mouse model of spinal metastasis of human prostate cancer using fluorescence-labeled PC-3 cells to allow direct observation by in vivo imaging. METHODS: PC-3 cells were infected with a lentivirus carrying green fluorescence protein (GFP) gene. The GFP-positive cell clone was expanded and prepared into cell suspension for injection into the inferior vena cava of nude mice. The tumor growth and metastasis in the mice was directly observed using an in vivo fluorescence imaging system. The tumor-bearing mice were sacrificed after 3 months for histological examination with HE staining. RESULTS: The labeled cells showed stable GFP expression both in vitro and in vivo. One week after cell injection, green fluorescence signals were detected by the in vivo fluorescence imaging system in the lower back of the mice, and at 4 weeks, the fluorescent tumor mass increased with a bone metastasis rate of 19% (3/16). Dissection of the mice at 3 months revealed lumbar tumor infiltration in 3 mice, showing a consistent result with in vivo fluorescence imaging. CONCLUSION: The nude mouse model of spinal bone metastasis of human prostate cancer established using GFP-labeled PC-3 cells facilitates further study of bone metastasis of prostate cancer.