The Role of m6A in Osteoporosis and the Differentiation of Mesenchymal Stem Cells into Osteoblasts and Adipocytes.

Osteoporosis is a systemic disease in which bone mass decreases, leading to an increased risk of bone fragility and fracture. The occurrence of osteoporosis is believed to be related to the disruption of the differentiation of mesenchymal stem cells into osteoblasts and adipocytes. N6-adenylate methylation (m6A) modification is the most common type of chemical RNA modification and refers to a methylation modification formed by the nitrogen atom at position 6 of adenine (A), which is catalyzed by a methyltransferase. The main roles of m6A are the post-transcriptional level regulation of the stability, localization, transportation, splicing, and translation of RNA; these are key elements of various biological activities, including osteoporosis and the differentiation of mesenchymal stem cells into osteoblasts and adipocytes. The main focus of this review is the role of m6A in these two biological processes.

Is the Zero-P Spacer Suitable for 3-Level Anterior Cervical Discectomy and Fusion Surgery in Terms of Sagittal Alignment Reconstruction: A Comparison Study with Traditional Plate and Cage System.

The Zero-P spacer was primarily developed aiming to reduce the morbidity associated with the traditional anterior cervical plate. During the past decade, many authors have reported the use of Zero-P spacers for anterior cervical discectomy and fusion (ACDF) of one or two segments. Nevertheless, there is still a paucity of knowledge on the safety and feasibility of using Zero-P spacers for 3-level fixation. The objective of this study was to investigate the clinical and radiological outcomes, with a focus on the sagittal alignment reconstruction of 3-level ACDF surgery using Zero-P spacers versus those using a traditional plate and cage system. From Sep 2013 to Aug 2016, a total of 44 patients who received 3-level ACDF surgery due to cervical spondylotic myelopathy were recruited. The Zero-P spacer was used in 23 patients (group ZP) and the traditional plate and cage system in 21 (group PC). Clinical outcomes were analyzed by Neck Disability Index (NDI) and Japanese Orthopedic Association (JOA) scores, and dysphagia was evaluated using the Bazaz score. Radiological outcomes, including fusion rate, adjacent segment degeneration (ASD), and especially changes in cervical sagittal alignment, were analyzed. The NDI and JOA scores did not differ significantly between the two groups postoperatively (p > 0.05); however, there was significantly less dysphagia in patients using Zero-P spacers at the 3- and 6-month follow-up (p < 0.05). At the 24-month follow-up, the fusion rate and ASD were similar between the two groups (p > 0.05). Interestingly, patients using Zero-P spacers had a significantly lower postoperative C2-7 Cobb angle and fused segment Cobb angle, compared to those using a traditional plate and cage system (p < 0.05); meanwhile, the fused segment disc wedge was also found to be significantly smaller in patients using Zero-P spacers after surgery (p < 0.05). Moreover, we further divided patients into subgroups according to their cervical lordosis. In patients with a preoperative C2-7 Cobb angle ≤ 10°, significantly less cervical and local lordosis, as well as disc wedge, were seen in group ZP after surgery (p < 0.05), while in others with a preoperative C2-7 Cobb angle > 10°, no significant difference in postoperative changes of the cervical sagittal alignment was seen between group ZP and group PC (p > 0.05). Zero-P spacers used in 3-level ACDF surgery could provide equivalent clinical outcomes and a lower rate of postoperative dysphagia, compared to the traditional plate and cage system. However, our results showed that it was inferior to the cervical plate in terms of sagittal alignment reconstruction for 3-level fixation. We recommend applying Zero-P spacers for 3-level ACDF in patients with good preoperative cervical lordosis (C2-7 Cobb angle > 10°), in order to restore and maintain physiological curvature of the cervical spine postoperatively.

[Comparison of short-term effectiveness of Prodisc-C Vivo artificial disc replacement and Zero-P fusion for treatment of single-segment cervical spondylosis].

Objective: To compare the short-term effectiveness and the impact on cervical segmental range of motion using Prodisc-C Vivo artificial disc replacement and Zero-P fusion for the treatment of single-segment cervical spondylosis. Methods: The clinical data of 56 patients with single-segment cervical spondylosis who met the selection criteria between January 2015 and December 2018 were retrospectively analyzed, and they were divided into study group (27 cases, using Prodisc-C Vivo artificial disc replacement) and control group (29 cases, using Zero-P fusion) according to different surgical methods. There was no significant difference between the two groups in terms of gender, age, type of cervical spondylosis, disease duration, involved segments and preoperative pain visual analogue scale (VAS) score, Japanese Orthopaedic Association (JOA) score, neck disability index (NDI), surgical segments range of motion, upper and lower adjacent segments range of motion, overall cervical spine range of motion, and cervical curvature (P>0.05). The operation time, intraoperative blood loss, postoperative hospitalization stay, time of returning to work, clinical effectiveness indicators (VAS score, JOA score, NDI, and improvement rate of each score), and imaging indicators (surgical segments range of motion, upper and lower adjacent segments range of motion, overall cervical spine range of motion, and cervical curvature, prosthesis position, bone absorption, heterotopic ossification, etc.) were recorded and compared between the two groups. Results: There was no significant difference in operation time and intraoperative blood loss between the two groups (P>0.05); the postoperative hospitalization stay and time of returning to work in the study group were significantly shorter than those in the control group (P<0.05). Both groups were followed up 12-64 months, with an average of 26 months. There was no complication such as limb or organ damage, implant failure, and severe degeneration of adjacent segments requiring reoperation. The VAS score, JOA score, and NDI of the two groups at each time point after operation significantly improved when compared with those before operation (P<0.05); there was no significant difference in the above scores at each time point after operation between the two groups (P>0.05); there was no significant difference in the improvement rate of each score between the two groups at last follow-up (P>0.05). The surgical segments range of motion in the study group maintained to varying degrees after operation, while it in the control group basically disappeared after operation, showing significant differences between the two groups (P<0.05). At last follow-up, there was no significant difference in the upper and lower adjacent segments range of motion in the study group when compared with preoperative ones (P>0.05), while the upper adjacent segments range of motion in the control group increased significantly (P<0.05). The overall cervical spine range of motion and cervical curvature of the two groups decreased at 3 months after operation, and increased to varying degrees at last follow-up, but there was no significant difference between groups and within groups (P>0.05). At last follow-up, X-ray films and CT examinations showed that no prosthesis loosening, subsidence, or displacement was found in all patients; there were 2 cases (7.4%) of periprosthetic bone resorption and 3 cases (11.1%) of heterotopic ossification which did not affect the surgical segments range of motion. Conclusion: Both the Prodisc-C Vivo artificial disc replacement and Zero-P fusion have satisfactory short-term effectiveness in treatment of single-segment cervical spondylosis. Prodisc-C Vivo artificial disc replacement can also maintain the cervical spine range of motion to a certain extent, while reducing the occurrence of excessive motion of adjacent segments after fusion.

Exosomes derived from microRNA-21 overexpressed adipose tissue-derived mesenchymal stem cells alleviate spine osteoporosis in ankylosing spondylitis mice.

MicroRNA-21 (miR-21) can induce proliferation and differentiation of mesenchymal stem cells (MSCs) to promote bone formation, we therefore aimed to investigate whether exosomes derived from miR-21 overexpressing adipose tissue-derived MSCs (AD-MSCs) could improve spine osteoporosis in ankylosing spondylitis (AS) mice. Cultured AD-MSCs were transfected with lentivirus vectors containing miR-21 or control vector, and the supernatant was centrifugated and filtrated to harvest the exosomes (miR-21-Exos or vector-Exos). BALB/c mice were immunized with cartilage proteoglycan to establish proteoglycan-induced ankylosing spondylitis (PGIA) model. Six weeks later, PGIA mice were further injected with miR-21-Exos or vector-Exos. Transfection of miR-21 in AD-MSCs significantly enhanced miR-21 levels in AD-MSCs and their exosomes. miR-21-Exos showed concentration-dependent protective effect against spine osteoporosis in PGIA mice, evidenced by increased bone mineral content and bone mineral density, reduced number of osteoclasts, decreased content of deoxypyridinoline in the urine, decreased content of tartrate-resistant acid phosphatase (TRACP)-5b and cathepsin K in the serum, and down-regulated interleukin (IL)-6 expression in the spine, whereas vector-Exos did not show any treatment benefit. The above findings indicate that miR-21-Exos could be utilized to treat spine osteoporosis in AS.

p53 inhibits CTR1-mediated cisplatin absorption by suppressing SP1 nuclear translocation in osteosarcoma.

Background: Osteosarcoma (OS) is a malignant bone tumor mainly affecting children and young adolescents. Cisplatin is a first-line chemotherapy drug for OS, however, drug resistance severely limits the survival of OS. Nevertheless, cellular factors in cisplatin resistance for OS remain obscure. In this study, the function and potential mechanism of p53 in cisplatin absorption were explored in OS cells. Methods: The CRISPR-Cas9 gene editing technology was performed to obtain p53 gene knock-out U2OS cells. The p53 over-expression 143B cell line was established by lentivirus-mediated virus infection. Moreover, the functions of p53 and CTR1 in cisplatin absorption were assessed by inductively coupled plasma mass spectrometry (ICP-MS) through CTR1 over-expression and knock-down. Further, the DNA binding activity of SP1 on CTR1 gene promoter was determined by dual-luciferase assay and chromatin immunoprecipitation (ChIP) assay. The functional regulation of p53 on SP1 was studied by nucleocytoplasmic separation assay and electrophoretic mobility shift assay (EMSA). The interaction between p53 and SP1 was verified by Co-Immunoprecipitation assay. Results: Under cisplatin treatment, p53 knock-out promoted CTR1 expression and cisplatin uptake, while p53 overexpression inhibited CTR1 expression and cisplatin uptake. Moreover, p53 regulated CTR1 level not by binding to CTR1 promoter directly but by suppressing the nuclear translocation of transcription factor specificity protein 1 (SP1). It was verified that SP1 is directly bound with CTR1 promoter. SP1 overexpression stimulated CTR1 expression, and SP1 knock-down attenuated CTR1 expression. Conclusion: The p53 might function as a negative regulator in CTR1 mediated cisplatin absorption, and the p53-SP1-CTR1 axis is a target for cisplatin resistance.

A Versatile Punch Stroke Correction Model for Trial V-Bending of Sheet Metals Based on Data-Driven Method.

During air bending of sheet metals, the correction of punch stroke for springback control is always implemented through repeated trial bending until achieving the forming accuracy of bending parts. In this study, a modelling method for correction of punch stroke is presented for guiding trial bending based on a data-driven technique. Firstly, the big data for the model are mainly generated from a large number of finite element simulations, considering many variables, e.g., material parameters, dimensions of V-dies and blanks, and processing parameters. Based on the big data, two punch stroke correction models are developed via neural network and dimensional analysis, respectively. The analytic comparison shows that the neural network model is more suitable for guiding trial bending of sheet metals than the dimensional analysis model, which has mechanical significance. The actual trial bending tests prove that the neural-network-based punch stroke correction model presents great versatility and accuracy in the guidance of trial bending, leading to a reduction in the number of trial bends and an improvement in the production efficiency of air bending.

Modified surgical treatment for a patient with neurofibromatosis scoliosis: A case report.

BACKGROUND: The correction surgery for severely multidimensional spinal deformity in neurofibromatosis type I is very difficult and it is still a very big challenge for spine surgeons. CASE SUMMARY: A 44-year-old woman presented with progressive kyphosis for more than 10 years and low back pain for 2 years. She had been diagnosed with neurofibromatosis at a local hospital many years ago. Conservative treatments had been applied, but the symptoms got worse rather than alleviated. Therefore, surgery was required. CONCLUSION: For this patient with severe deformity, the correction treatment of Ponte osteotomy followed by satellite rod technique in the region of the apical vertebra and the technique of pedicle screws and dual iliac screws had been applied, and successful clinical outcomes were achieved.

Effect of Deep Cryogenic Activated Treatment on Hemp Stem-Derived Carbon Used as Anode for Lithium-Ion Batteries.

The cryogenic process has been widely applied in various fields, but it has rarely been reported in the preparation of anode materials for lithium-ion battery. In this paper, activated carbon derived from hemp stems was prepared by carbonization and activation; then, it was subjected to cryogenic treatment to obtain cryogenic activated carbon. The characterization results show that the cryogenic activated carbon (CAC) has a richer pore structure than the activated carbon (AC) without cryogenic treatment, and its specific surface area is 1727.96 m2/g. The porous carbon had an excellent reversible capacity of 756.8 mAh/g after 100 cycles at 0.2 C as anode of lithium-ion battery, in which the electrochemical performance of CAC was remarkably improved due to its good pore structure. This provides a new idea for the preparation of anode materials for high-capacity lithium-ion batteries.

Circular RNA hsa_circ_0076906 competes with OGN for miR-1305 biding site to alleviate the progression of osteoporosis.

Osteoporosis is a skeletal disorder, the pathogenic factors of which include the decreased ability of osteogenesis and enhanced osteolysis. Human-derived mesenchymal stem cells (hMSCs) possess the differential capacity to osteoblasts and chondrocytes, so the application of hMSCs in bone tissue is promising to be effective. A group of RNAs which lack of 5' and 3'ends called circular RNAs (circRNAs) were discovered. In this study, we described a previously found circular RNA, circ_0076906, to bind miR-1305 and regulate its target gene, Osteoglycin (OGN), thus regulate osteogenic differentiation of hMSCs and alleviate the progression of osteoporosis. Osteogenic differentiation induced in hMSCs; qRT-PCR and western blot to examine the expressions of mRNAs and proteins; Alkaline phosphatase activity and Alizarin red staining to examine bone formation; luciferase report experiments to detect the interaction between molecules; and the nuclear/cytoplasm separation of cells. 1. Circ_0076906 was induced in osteogenic differentiation; 2. Circ_0076906 silencing inhibited osteogenesis-related genes in hMSCs; 3. Circ_0076906 acted as a sponge for miR-1305; 4. MiR-1305 regulated OGN expression; 5. Circ_0076906 induced-osteogenic differentiation depended on miR-1305/ OGN pathway. Circ_0076906 relieved osteoporosis and promoted osteogenic differentiation through the miR-1305/ OGN pathway.

Characterization and Preparation of Nano-porous Carbon Derived from Hemp Stems as Anode for Lithium-Ion Batteries.

As a biomass waste, hemp stems have the advantages of low cost and abundance, and it is regarded as a promising anode material with a high specific capacity. In this paper, activated carbon derived from hemp stems is prepared by low-temperature carbonization and high-temperature activation. The results of characterizations show the activated carbon has more pores due to the advantages of natural porous structure of hemp stem. The aperture size is mainly microporous, and there are mesopores and macropores in the porous carbon. The porous carbon has an excellent reversible capacity of 495 mAh/g after 100 cycles at 0.2 °C as the anode of lithium-ion battery. Compared with the graphite electrode, the electrochemical property of activated carbon is significantly improved due to the reasonable distribution of pore size. The preparation of the activated carbon provides a new idea for low cost and rapid preparation of anode materials for high capacity lithium-ion batteries.

rna interference targeting p110ß reduces tumor necrosis factor-alpha production in cellular response to wear particles in vitro and osteolysis in vivo.

Aseptic joint loosening is a key factor that reduces the life span of arthroplasty. There are currently few effective treatments for joint loosening except surgical revision. We explored the inhibitory effects of p110ß-targeted small interfering RNA (siRNA) and lentivirus on particle-induced inflammatory cytokine expression in the murine macrophage cell line, RAW264.7. siRNA- and lentivirus-targeting p110ß were transfected and infected prior to particle stimulation, respectively. Ceramic and titanium particles of different sizes were prepared to stimulate macrophages. Fluorescence microscopy showed that the efficiency of siRNA transfection and lentivirus infection were 74.2 ± 4.2 and 92.3 ± 2.6 %, respectively. TNF-alpha mRNA in the particle stimulation plus RNA interference (RNAi) groups were significantly lower compared with the particle stimulation-only groups (P < 0.05). Similarly, protein levels of TNF-alpha in RNAi-treated groups were significantly decreased after transfection or infection (P < 0.05). It showed that Phosphor-AKT (Ser473) activation was significantly reduced by RNAi through western blot. As assessed by CT, micro-CT and histological analysis, particle implantation induced a significant osteolysis in mice calvaria, which was limited by p110ß lentivirus addition. These results suggested that p110ß subtype of PI3K, followed by activation of Ser473, may possibly participate in the regulation of macrophages activity by wear particles, ultimately resulting in the TNF-α secretion and osteolysis.