CGRP promotes osteogenic differentiation by regulating macrophage M2 polarization through HDAC6/AKAP12 signaling pathway.

Aim: To determine the mechanism of Calcitonin gene-related peptide (CGRP) in bone healing. Materials & methods: Alkaline phosphatase (ALP) activity and inflammatory-factor levels were detected using ELISA. Osteogenic differentiation was assessed using Alizarin red staining technique. The interaction between histone deacetylase 6 (HDAC6) and A-kinase anchoring protein 12 (AKAP12) was investigated through Co- immunoprecipitation. Results: CGRP treatment promoted rat bone marrow-derived macrophages (BMDMs) M2 polarization. CGRP facilitated osteogenic differentiation by enhancing M2 polarization of BMDMs. Mechanistically, CGRP promoted AKAP12 acetylation to activate the extracellular regulated protein kinases pathway by HDAC6 inhibition. Conclusion: CGRP promoted M2 polarization of rat BMDMs and facilitated osteogenic differentiation through the HDAC6/AKAP12/extracellular regulated protein kinases signaling pathway, thereby promoting bone healing.

[Box: see text].

Research on driver's anger recognition method based on multimodal data fusion.

OBJECTIVES: This paper aims to address the challenge of low accuracy in single-modal driver anger recognition by introducing a multimodal driver anger recognition model. The primary objective is to develop a multimodal fusion recognition method for identifying driver anger, focusing on electrocardiographic (ECG) signals and driving behavior signals. METHODS: Emotion-inducing experiments were performed employing a driving simulator to capture both ECG signals and driving behavioral signals from drivers experiencing both angry and calm moods. An analysis of characteristic relationships and feature extraction was conducted on ECG signals and driving behavior signals related to driving anger. Seventeen effective feature indicators for recognizing driving anger were chosen to construct a dataset for driver anger. A binary classification model for recognizing driving anger was developed utilizing the Support Vector Machine (SVM) algorithm. RESULTS: Multimodal fusion demonstrated significant advantages over single-modal approaches in emotion recognition. The SVM-DS model using decision-level fusion had the highest accuracy of 84.75%. Compared with the driver anger emotion recognition model based on unimodal ECG features, unimodal driving behavior features, and multimodal feature layer fusion, the accuracy increased by 9.10%, 4.15%, and 0.8%, respectively. CONCLUSIONS: The proposed multimodal recognition model, incorporating ECG and driving behavior signals, effectively identifies driving anger. The research results provide theoretical and technical support for the establishment of a driver anger system.

A Study on the Effect of Precise Rehabilitation Therapy Guided by Three-dimensional-computed Tomography Reconstruction Technology in Hip Fracture Surgery Patients.

OBJECTIVE: To evaluate the effectiveness of precise rehabilitation therapy guided by three-dimensional computed tomography (3D-CT) reconstruction technology in hip fracture patients through a retrospective cohort study. METHOD: Data were retrospectively collected from 60 patients aged over 60 who had undergone hip fracture surgery. They were divided into two groups based on their chosen rehabilitation method: a control group and a test group. The study collected demographic data, fracture characteristics, and quality of life indicators to assess the impact of rehabilitation on economic indicators and daily living activities (ADL). Additionally, it included assessments of muscle strength, joint mobility, hip function, postoperative complications, and records of hospitalization information and costs. Cognitive function was also assessed postoperatively. RESULTS: There were no significant differences in demographic data, fracture characteristics, ADL, or Fugl-Meyer assessment (FMA) between the two groups. However, the test group exhibited significantly higher post-surgery muscle strength recovery and hip mobility compared to the control group (P<0.05). Additionally, the test group had significantly fewer hospitalization days and lower hospitalization costs than the control group (P<0.05). CONCLUSION: Precise rehabilitation therapy guided by 3D-CT reconstruction technology for hip fracture surgery patients can enhance early muscle strength recovery, improve mobility of the affected limb, reduce hospitalization duration and costs, and enhance overall patient recovery outcomes.

Repairing of Subchondral Defect and Articular Cartilage of Knee Joint of Rabbit by Gadolinium Containing Bio-Nanocomposites.

A variety of gadolinium (Gd) based nanoparticles (NPs) were synthesized due to the unique magnetic properties of Gd-containing rare earth compounds and the particularity of micro/nano-materials, which were then incorporated into hydroxyapatite (HA) to obtain inorganic-organic composite materials. Then, HA/Gd NPs containing slow-release transforming growth factor (TGF-ß1) were harvested. Adipose-derived stem cells (ADSCs) were extracted from the adipose tissue of a four-month-old New Zealand white rabbit. HA/Gd NPs were attached to absorbable gelatin sponge to obtain HA/Gd NPs/gelatin sponge composite scaffold. In addition, the third generation ADSCs were taken and cultured in the composite scaffold, so that ADSCs-HA/Gd bio-nanocomposites were obtained. The in vitro culture test of osteoblast MC3T3-E1 showed that Gd-containing NPs had good biocompatibility. The prepared HA/Gd NPs loaded with TGF-ß1 were spherical, with an average particle size of (9.16 ± 3.16) µm. The NPs were easy to aggregate and adherent. Enzyme-linked immunosorbent assay (ELISA) test results showed that TGF-ß1 in NPs was sustained and released continuously for 29 days. HA/Gd NPs/gelatin sponge composite scaffold combined with ADSCs were co-cultured for three days, and the electron microscope showed that the HA/Gd NPs were dispersed, and the cells could adhere and grow well. Then, animal models of rabbit knee articular cartilage defects were established and were rolled into three groups (ADSCs-HA/Gd nano group, HA/Gd nano scaffold group, and blank control). The repair area of the rabbit knee of ADSCs-HA/Gd nano group was smooth and flat, the scaffold was absorbed, the toluidine blue stain was positive, and the type II collagen immunohistochemical stain was positive. In general, ADSCs-HA/Gd nanomaterials were helpful for chondrogenic cell differentiation and had certain adoption prospects in the field of tissue engineering to repair cartilage defects.

Puerarin facilitates osteogenesis in steroid-induced necrosis of rabbit femoral head and osteogenesis of steroid-induced osteocytes via miR-34a upregulation.

The present study investigated the effect of puerarin on promoting the osteogenesis in steroid-induced necrosis of the femoral head (SONFH). New Zealand rabbits were administrated with horse serum and methylprednisolone (MPS) for establishing SONFH in vivo model, which was then treated with puerarin treatment. Histo-morphological changes in the femoral head were examined by hematoxylin-eosin staining. Osteoblasts were isolated from healthy rabbits and treated by individual or combined administration of dexamethasone and puerarin. Osteoblast viability was measured by CCK-8 assay. Mineralized nodule formation was evaluated by alizarin red assay. Expressions of RUNX family transcription factor 2 (RUNX2), Type-I collagen α 1 (COL1A1), ALP and miR-34a in the femoral head were determined by qRT-PCR and Western blot. Puerarin attenuated the effect of SONFH on promoting histopathological abnormalities and counteracted SONFH inhibition on the expressions of ALP, RUNX2, COL1A1 and miR-34a in the rabbits. Rabbit osteoblasts were successfully isolated, as they showed red mineralized nodules. Dexamethasone exposure decreased osteoblast viability, which was increased by puerarin treatment. Furthermore, puerarin treatment attenuated dexamethasone-induced inhibition on the viability, osteoblastic differentiation, and the expressions of ALP, RUNX2, COL1A1 and miR-34a in the osteoblasts. Puerarin facilitated osteogenesis of steroid-induced necrosis of rabbit femoral head and osteogenesis of steroid-induced osteocytes via miR-34a upregulation.

MiR-214 regulates fracture healing through inhibiting Sox4 and its mechanism.

OBJECTIVE: To investigate the expression of micro ribonucleic acid (miR)-214 in the bone tissue and blood of patients with fragility fracture. METHODS: The expression of miR-214 was detected via quantitative reverse transcription-polymerase chain reaction. The effect of miR-214 on proliferation and apoptosis of osteoblasts were detected via methyl thiazolyl tetrazolium assay and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling staining. RESULTS: The expression of miR-214 in the bone tissue and blood of patients with fragility fracture significantly declined. miR-214 could promote the proliferation of osteoblasts and inhibited the apoptosis of osteoblasts. miR-214 is involved in fracture healing through inhibiting Sox4 and promoting phosphorylation of PI3K/AKT pathway. The expression of BSP in cells treated with miR-214 mimics was significantly increased to 2.5-fold (p=0.0168), while the expression of BSP in cells treated with miR-214 AMO was significantly decreased, reduced to 0.3 times (p=0.0397). The expression of BMP2 in cells treated with miR-214 mimics was significantly increased to 2.5-fold (p=0.003), while the expression of BMP2 was significantly decreased in cells treated with miR-214 AMO, reduced to 0.3 times (p=0.0002). miR-214 can regulate the expression of Sox2, PI3K and AKT proteins. CONCLUSION: MiR-214 regulates the proliferation, apoptosis, bone formation of osteoblasts and participate in the fracture healing process by inhibiting the expression of Sox4, which provided new ideas for clinical treatment of fracture healing.

Hepatoprotective effects of Di Wu Yang Gan: A medicinal food against CCl4-induced hepatotoxicity in vivo and in vitro.

The development of functional foods based on medicinal food ingredients has become a hot topic in China. Di Wu Yang Gan (DWYG) is a Chinese medicinal food that contains five dietary plants. Various health benefits, including anti-inflammation, liver regeneration regulation, have been reported, though the mechanism is not clear. This study aimed to investigate the protective effect of DWYG on carbon tetrachloride-induced acute liver injury (ALI) in embryonic liver L-02 cells and mice model. DWYG-medicated serum protected L-02 cells from carbon tetrachloride-induced damage, reduced the levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in the culture medium, decreased the expression of Bax and increased the expression of Bcl-2. Mice study suggested that DWYG decreased the levels of malondialdehyde, ALT and AST. Together, these results suggest the hepatoprotective effects of DWYG against ALI and provide an experimental basis for the utilization of DWYG to treat liver damage.

Pinoresinol promotes MC3T3­E1 cell proliferation and differentiation via the cyclic AMP/protein kinase A signaling pathway.

Estradiol (E2) is a first­line drug for osteoporosis (OP) treatment via promotion of osteoblastic proliferation and differentiation. However, a long­term use of E2 would produce side effects thus, it is imperative to discover safer and more effective drugs. Pinoresinol (PINO) has a similar chemical structure to E2. The present study aimed to investigate whether PINO could promote osteoblastic proliferation and differentiation and the potential mechanisms. After treatment with 0.1 µg/l PINO for 2 days, MC3T3­E1 cell migration was assessed by wound healing assay. Estrogen (E2) treatment served as a positive control. RT­qPCR and western blotting were used for mRNA and protein expression analyses. Alkaline phosphatase (ALP) activity assay and Alizarin red staining were performed to investigate the calcification and mineralization, and the cyclic AMP (cAMP) level was detected by enzyme­linked immunosorbent assay (ELISA). H89, an inhibitor of protein kinase A (PKA), was introduced to verify the role of cAMP/PKA in the effect of PINO on MC3T3­E1 cells. Cell viability was the highest under 48 h of 0.1 µg/l PINO treatment. After treatment with PINO, a significant increase was observed in the migration rate and the expression of collagen type I (Col­I), ALP, osteopontin (OPN), runt­related transcription factor 2 (Runx2) and bone morphogenetic protein­2 (BMP­2) (P<0.01). The ALP activity and Alizarin red size in PINO and E2 groups were notably increased. The increased cAMP, PKA and phosphorylated cAMP response element­binding protein (CREB) levels were also observed in the PINO group. Furthermore, H89 co­treatment abolished the positive effects of PINO on cell viability and migration. PINO had similar effects to E2 on the osteoblastic proliferation and differentiation, and these positive effects may be attributed to the regulation of the cAMP/PKA signaling pathway.

miR-4262 regulates chondrocyte viability, apoptosis, autophagy by targeting SIRT1 and activating PI3K/AKT/mTOR signaling pathway in rats with osteoarthritis.

The present study aimed to investigate the effect and underlying mechanism of microRNA (miR)-4262 in the development of osteoarthritis (OA) in rats. Primary chondrocytes were separated from Sprague-Dawley rats and then treated with tumor necrosis factor-α (TNF-α). The level of miR-4262 was detected in TNF-α-treated chondrocytes, and then the miR-4262 or its target gene sirtuin type 1 (SIRT1) level was overexpressed, or knocked down. Furthermore, cell viability, cell apoptosis, cell autophagy and matrix synthesis, as well as the expressions of proteins associated with the phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT)/mammalian target of rapamycin (mTOR) signaling pathway were detected. miR-4262 was significantly overexpressed in TNF-α-treated chondrocytes compared with untreated cells (P<0.05). TNF-α treatment or miR-4262 overexpression significantly decreased cell viability, autophagy-related proteins levels and matrix synthesis-related proteins levels, as well as increased the apoptotic rate in chondrocytes (P<0.05). Overexpression of SIRT1 significantly increased cell viability, autophagy-related proteins levels and matrix synthesis-related proteins levels, as well as decreased the apoptotic rate in TNF-α-treated chondrocytes (P<0.05). In addition, the effects of miR-4262 on cell viability, cell apoptosis, cell autophagy and matrix synthesis were inhibited by SIRT1 (P<0.05). Furthermore, upregulated miR-4262 remarkably increased the expressions of phosphorylated (p)-PI3K, p-AKT and p-mTOR (P<0.05) in TNF-α treated chondrocytes. The present study revealed that the upregulation of miR-4262 may promote the occurrence and development of OA in rats by regulating cell viability, cell apoptosis, cell autophagy, and matrix synthesis. Furthermore, these roles of miR-4262 may be associated with PI3K/AKT/mTOR signaling pathway.