The anterior talofibular ligament: A thin-slice three-dimensional magnetic resonance imaging study.

PURPOSE: The aim of this study was to provide an accurate and improved understanding of anterior talofibular ligament (ATFL) anatomy, and to determine the exact positioning and diameter of the bony tunnel during ATFL repair and/or reconstruction surgery. METHOD: A total of 58 healthy asymptomatic volunteers were examined, wherein 38 underwent bilateral ankle 3D MRI, and 20 underwent unilateral ankle 3D MRI (10 left and 10 right ankles). Data from a total of 96 MRI datasets were collected. The MRI data from these cases were exported into Mimics to enable reconstruction of 3D ATFL models. The resulting image quality was evaluated using a 5-point subjective scoring system. In addition, the length, width, thickness, and positioning of each ATFL and the area of the ATFL footprints were identified within the 3D model using Mimics and SolidWorks. RESULTS: The image quality score was 4.48 ± 0.50. The ATFL formed one (65.6%), two (31.3%), or three (3.1%) bundles forms. The footprint area was 31.25 ± 6.29 mm2 on the fibular side, and 17.48 ± 4.49 mm2 on the talar side. CONCLUSION: Thin-slice 3D MRI aids in the reconstruction of the 3D ATFL model, and it provides reference for the accurate anatomy of the area and location of the ATFL. This technology will facilitate diagnosis of ATFL injuries and choice of surgical methods. LEVEL OF EVIDENCE: level IV.

Evaluation of 3-Dimensional Magnetic Resonance Imaging (3D MRI) in Diagnosing Anterior Talofibular Ligament Injury.

BACKGROUND It is challenging to entirely show the anterior talofibular ligament (ATFL) and accurately diagnose ATFL injury with traditional 2-dimensional (2D) magnetic resonance imaging (MRI). With the introduction of 3.0T MRI, a 3-dimensional (3D) MRI sequence can achieve images with high spatial resolution. This study aimed to evaluate the accuracy of 3D MRI and compare it with 2D MRI in diagnosing ATFL injury. MATERIAL AND METHODS This was a prospective study in which 45 patients with clinically suspected ATFL injury underwent 2D MRI, 3D MRI, and 3D model reconstruction followed by arthroscopic surgery between February 2018 and April 2019. Two radiologists who had over 11 and 13 years of musculoskeletal experience assessed the injury of ATFL in consensus without any clinical clues. Arthroscopic surgery results were the standard reference of MRI accuracy. RESULTS The 3D MRI results of ATFL injury showed the sensitivity of diagnosis of complete tears of 83% and specificity of 82%. The partial tears diagnosis sensitivity was 78%, and specificity was 100%. The sensitivity of diagnosis of sprains was 100%, and the specificity was 97%. The 3D MRI accuracy of diagnosis was 98% for no injury, 98% for sprain, 91% for partial tear, and 82% for complete tear. The difference in the diagnosis of sprain and partial tears by 3D MRI and 2D MRI was statistically significant (P<0.05). A 3D reconstruction model was successfully created for all patients. CONCLUSIONS 3D MRI may be a reliable and accurate method to detect ATFL injury. The 3D reconstruction model using 3D MRI sequences has excellent prospects in application.

Inhibition of exosomal miR-24-3p in diabetes restores angiogenesis and facilitates wound repair via targeting PIK3R3.

Diabetic foot ulcer (DFU) is one of the common ailments of elderly people suffering from diabetes. Exosomes containing various active regulators have been found to play a significant role in apoptosis, cell proliferation and other biological processes. However, the effect and the underlying mechanism of action of diabetes patients derived from circulating exosomes (Dia-Exos) on DFU remain unclear. Herein, we aim to explore the potential regulatory role of Dia-Exos. First, we attempted to demonstrate the harmful effect of Dia-Exos both in vivo and in vitro. miRNA-24-3p (miR-24-3p) was found enriched with Dia-Exos. Hence, inhibition of this miRNA could partially reverse the negative effect of Dia-Exos, thus, in ture, accelerates wound repair. Luciferase assay further verified the binding of miR-24-3p to the 3'-UTR of phosphatidylinositol 3-kinase regulatory subunit gamma (PIK3R3) mRNA and the PIK3R3 expression enhanced human umbilical vein endothelial cells functionality in vitro. Hence, the findings of this study reveal the regulatory role of Dia-Exos in the process of wound healing and provide experimental evidence for the therapeutic effects of knocking down miR-24-3p in DFU treatment.

Therapeutic effect of omega-3 fatty acids on T cell-mediated autoimmune diseases.

The present study was to demonstrate that the G protein coupled receptors serve as targets for the treatment of autoimmune disease such as rheumatoid arthritis and multiple sclerosis. Rats received pristane at the base of the tail. Affected joints were counted daily. The T cell mediated autoimmune diseases such as pristine-induced arthritis (PIA) and autoimmune encephalomyelitis (EAE) in a rat model were profoundly ameliorated by treatment with the specific G protein couple receptors 120 (GPR120) stimuli omega-3 fatty acids (ω-3 FAs). Our study further revealed that the activation of GPR120 by ω-3 FAs can result in a decrease of phosphorylated transforming growth factor-ß activated kinase 1 (TAK1), and further inhibit the downstream IKKß/I-κB pathway and the terminal NF-κB activation which serves as a mediator of T cell activation. ω-3 Fatty acids exhibited an inhibitory effect on TAK1 by enhancing the association of ß-arrestin2 and TAK1 binding protein 1 (TAB1), thus the disassociation of TAB1 from the TAB1/TAK1 complex renders a limited effect on ß-arrestin2 signaling as an innate immunity regulation. GPR120 is a functional receptor of ω-3 fatty acids in T cell-mediated autoimmune disease compared with its effect on innate immunity.

Suture Anchor Versus Allogenic Tendon Suture in Treatment of Haglund Syndrome.

BACKGROUND Haglund's deformity is an abnormal bony enlargement on the back of the heel. It can cause the impact of the posterior calcaneal bursa and Achilles tendon insertion, and finally result in pain. This syndrome is called Haglund syndrome. The purpose of this study was to explore the effect of the suture anchor and allogeneic tendon suture in the treatment of Haglund syndrome. MATERIAL AND METHODS We retrospectively studied 20 patients with Haglund syndrome treated from January 2015 to December 2016. The patients were randomly divided into Group 1 (the suture anchor group) and Group 2 (the allogeneic tendon group), with 10 patients in each group and an average follow-up of 32 months after surgery. The AOFAS, VAS, and Arner-Lindholm scales were used to summarize the patient follow-up results and complications. RESULTS In the 2 groups of patients, the postoperative AOFAS, VAS scores, and the Arner-Lindholm scale showed good results. However, the postoperative AOFAS score and VAS of the suture anchor group were better than those of the allogeneic tendon group, with shorter operation times. No Achilles tendon rupture or wound infection occurred during the entire postoperative period in either group. These results show the superiority of suture anchors. CONCLUSIONS The higher AOFAS and VAS score and shorter operation time in the suture anchor group suggest it is the better alternative for treatment of Haglund syndrome.

Investigation of the Relationship Between Flatfoot and Patellar Subluxation in Adolescents.

Patellar subluxation is common in adolescents, and a variety of factors are related to this condition, with valgus of the knee joint an important factor. The results of many studies suggest that flatfoot can cause an abnormality of the lower limb power line. Structural abnormalities of the foot caused by the high stresses exerted by body weight can lead to structural deformity of the knee and can also cause knee valgus. Screening for foot problems can help determine the risk of patellar subluxation, and early intervention can lessen the incidence of this condition. The purpose of the present study was to investigate the effects of flatfoot on the structure and function of the knees and, especially, the risk of patellar subluxation. A total of 72 participants were recruited for this cross-sectional study. The mean age at examination was 15.4 ± 4.0 (range 9 to 22) years. The measured parameters were heel valgus angle, arch index, and quadriceps angle (Q-angle). Overall, the mean values of the heel valgus angle, arch index, and Q-angle were 5.9° ± 2.4° (range 1° to 11°), 0.33 ± 0.07 (range 0.23 to 0.46), and 19.1° ± 3.5° (range 9° to 26°), respectively. The Q-angle was directly associated with the heel valgus angle (r = 0.818, p < .001) and arch index (r = 0.655, p < .001). We found that flatfoot can affect the morphology of the knee joint and increase the risk of patellar subluxation.

Outcomes of arthroscopic arthrolysis for the post-traumatic elbow stiffness.

PURPOSE: Post-traumatic contracture is a common complication after elbow trauma. If conservative therapy fails to restore adequate elbow motion, surgical release is recommended. Increase in arthroscopy knowledge and skills, as well as technological advances in the passed decade of years, has made arthroscopic arthrolysis a safe and reliable treatment for patients with a post-traumatic elbow contracture. The aim of this study was to report on the clinical outcome and improvement of ROM in post-traumatic stiff elbow treated by arthroscopic arthrolysis. METHODS: Between 2008 and 2012, 34 consecutive patients with post-traumatic stiffness were treated with arthroscopic arthrolysis. Active and passive elbow movement is encouraged the day after operation with the effective pain management. Mayo Elbow Performance Index (MEPI) and visual analogue scale were measured. RESULTS: At the final follow-up, the average arc of elbow motion increased from 48.6 ± 19.3 pre-operatively to 114.5 ± 25.7, with a mean improvement of 65.9°. The MEPI score improved from 68.2 ± 16.4 pre-operatively to 92.4 ± 21.6, with a mean improvement of 24.2 (p < 0.001). Results were good to excellent in 29 patients. CONCLUSION: Injuries are the most common cause of elbow stiffness requiring surgical release. The procedure of arthroscopic arthrolysis is a good option for the treatment of post-traumatic elbow stiffness as it restores normal elbow function. Early passive/active post-operative rehabilitation is very important.

Functional outcome of limb-salvage surgery with shoulder abduction brace for bone tumors around the shoulders.

BACKGROUND: The current trend is toward salvage of the extremity after tumor excision without compromising the extent of resection for bone tumor around the shoulders. OBJECTIVES: The aim of this study was to evaluate functional outcome of patients treated with limb-salvage surgeries combined with shoulder abduction braces. METHODS: Thirty-six patients with bone tumors around the shoulders, who had limb-sparing resection and reconstruction performed with a shoulder abduction brace, were retrospectively reviewed. Allograft transplantation and rigid internal fixation was performed in 22 patients and artificial prosthetic replacement was performed in 14 patients. Functional evaluation was performed based on the Musculoskeletal Tumour Society (MSTS) scoring system. RESULTS: The overall survival was 78.8% (26/33) at 2 years. The mean final functional score was (81.2 ± 19.6%). The MSTS of patients treated by allograft transplantation and prosthetic replacement were (79.4 ± 15.3%) and (81.9 ± 18.1%), respectively. The MSTS scores differed only slightly between these two groups (P > 0.05). All the patients regained good ROM of the shoulder joints. CONCLUSIONS: Satisfactory functional outcomes can be obtained by limb-salvage surgery for bone tumor around the shoulder. Postoperatively shoulder crutches with shoulder abduction brace are encouraged as the aid of reconstruction of shoulder joint function.

Identifying interactive effects of spatial drivers in soil heavy metal pollutants using interpretable machine learning models.

The accurate identification of spatial drivers is crucial for effectively managing soil heavy metals (SHM). However, understanding the complex and diverse spatial drivers of SHM and their interactive effects remains a significant challenge. In this study, we present a comprehensive analysis framework that integrates Geodetector, CatBoost, and SHapley Additive exPlanations (SHAP) techniques to identify and elucidate the interactive effects of spatial drivers in SHM within the Pearl River Delta (PRD) region of China. Our investigation incorporated fourteen environmental factors and focused on the pollution levels of three prominent heavy metals: Hg, Cd, and Zn. These findings provide several key insights: (1) The distribution of SHM is influenced by the combined effects of various individual factors and interactions within the source-flow-sink process. (2) Compared with the spatial interpretation of individual factors, the interaction between Hg and Cd exhibited enhanced spatial explanatory power. Similarly, interactions involving Zn mainly demonstrated increased spatial explanatory power, but there was one exception in which a weakening was observed. (3) Spatial heterogeneity plays a crucial role in determining the contributions of environmental factors to soil heavy metal concentrations. Although individual factors generally promote metal accumulation, their effects fluctuate when interactions are considered. (4) The SHAP interpretable method effectively addresses the limitations associated with machine-learning models by providing understandable insights into heavy metal pollution. This enables a comparison of the importance of environmental factors and elucidates their directional impacts, thereby aiding in the understanding of interaction mechanisms. The methods and findings presented in this study offer valuable insights into the spatial heterogeneity of heavy metal pollution in soil. By focusing on the effects of interactive factors, we aimed to develop more accurate strategies for managing SHM pollution.

G-CSF/SCF exert beneficial effects via anti-apoptosis in rabbits with steroid-associated osteonecrosis.

OBJECTIVES: Osteonecrosis is also known as avascular necrosis, and two types of cell death are included in the pathogenesis of osteonecrosis: necrosis and apoptosis. Apoptosis in the osteonecrosis of femoral head is thought to be the key determinant of glucocorticoid-induced cortical bone loss. The present study was implemented to evaluate the anti-apoptotic effect of Granulocyte colony-stimulating factor and stem cell factor (G-CSF/SCF) in rabbits with steroid-induced osteonecrosis. METHODS: In the experiment, osteonecrosis was induced by low-dose lipopolysaccharide and subsequent pulsed high-dose methylprednisolone. Rabbits in preventive medicine group were treated with 100 µg/kg/d G-CSF and 25 µg/kg/d SCF. Then hematological and histomorphometric methods were used to investigate the treatment effects of osteonecrosis. Apoptosis was assessed via quantitative TUNEL staining and activated caspase-3 immunostaining and immunoblotting. RESULTS: The results showed that G-CSF/SCF treatment could increase the secretion of serum osteocalcin, but inhibit the expression of serum tartrate-resistant acid phosphatase (TRAP5b). The incidence of osteonecrosis was significantly decreased in Preventive group when compared with Steroid group (42.1% vs. 88.2%). Histomorphometric analysis showed that G-CSF/SCF pre-disposal treatment was able to increase trabecular mineral appositional rate (MAR) and bone formation rate (BFR). Quantitative TUNEL and activated caspase-3 levels showed lower apoptosis in the Preventive group. CONCLUSIONS: In conclusion, G-CSF/SCF treatment could inhibit caspase-3-dependent apoptosis in osteocytes to exert beneficial effects in preventing steroid-induced ON in rabbit models.

Copper Ion-Modified Germanium Phosphorus Nanosheets Integrated with an Electroactive and Biodegradable Hydrogel for Neuro-Vascularized Bone Regeneration.

Severe bone defects accompanied by vascular and peripheral nerve injuries represent a huge orthopedic challenge and are often accompanied by the risk of infection. Thus, biomaterials with antibacterial and neurovascular regeneration properties are highly desirable. Here, a newly designed biohybrid biodegradable hydrogel (GelMA) containing copper ion-modified germanium-phosphorus (GeP) nanosheets, which act as neuro-vascular regeneration and antibacterial agents, is designed. The copper ion modification process serves to improve the stability of the GeP nanosheets and offers a platform for the sustained release of bioactive ions. Study findings show that GelMA/GeP@Cu has effective antibacterial properties. The integrated hydrogel can significantly boost the osteogenic differentiation of bone marrow mesenchymal stem cells, facilitate angiogenesis in human umbilical vein endothelial cells, and up-regulate neural differentiation-related proteins in neural stem cells in vitro. In vivo, in the rat calvarial bone defect mode, the GelMA/GeP@Cu hydrogel is found to enhance angiogenesis and neurogenesis, eventually contributing to bone regeneration. These findings indicate that in the field of bone tissue engineering, GelMA/GeP@Cu can serve as a valuable biomaterial for neuro-vascularized bone regeneration and infection prevention.

Morphologic evaluation of injured and contralateral uninjured ankles in patients with unilateral chronic ankle instability.

OBJECTIVE: To compare the morphological anatomy and abnormalities of the anterior talofibular ligament (ATFL) and calcaneofibular ligament (CFL) in unilateral chronic ankle instability (CAI). METHODS: 22 patients (men: women, 13:9; mean age, 28.95 ± 8.127 years) with unilateral CAI and 18 healthy volunteers (men: women, 9:9, mean age, 28.33 ± 3.678 years) were recruited. MRI scans were divided into Group 1 (22 injured ankles), Group 2 (22 contralateral uninjured ankles), and Group 3 (36 healthy volunteer ankles). The morphologic variables, MRI signal intensity (SI) values were evaluated. RESULTS: The ATFL proximal, intermediate, and distal sites and the CFL proximal and distal sites in Group 3 were narrower than those in Group 1 (P ＜0.05). Both ATFL and CFL in Group 1 were thicker than those in Group 3 (P ＜0.01). The proximal and intermediate sites of the ATFL and the proximal site of the CFL in Group 3 were narrower than those in Group 2 (P ＜0.01). The intermediate site of the ATFL and the proximal and distal sites of the CFL in Group 2 were thicker than those in Group 3 (P ＜0.01). The mean SI values of the ATFL in Group 1 were higher than those in Groups 2 and 3 (P ＜0.01). The ATFL and CFL SI values were higher in Group 2 than those in Group 3 (P ＜0.05). CONCLUSION: Both the injured and contralateral uninjured ankles had wider ATFL and CFL, more thickness, and higher SI values compared with those of healthy volunteer ankles. ADVANCES IN KNOWLEDGE: High-resolution three-dimensional MRI provides a potential tool assisting clinical decision on the treatment and rehabilitation therapy of patients with unilateral CAI.

Stratified-structural hydrogel incorporated with magnesium-ion-modified black phosphorus nanosheets for promoting neuro-vascularized bone regeneration.

Angiogenesis and neurogenesis play irreplaceable roles in bone repair. Although biomaterial implantation that mimics native skeletal tissue is extensively studied, the nerve-vascular network reconstruction is neglected in the design of biomaterials. Our goal here is to establish a periosteum-simulating bilayer hydrogel and explore the efficiency of bone repair via enhancement of angiogenesis and neurogenesis. In this contribution, we designed a bilayer hydrogel platform incorporated with magnesium-ion-modified black phosphorus (BP) nanosheets for promoting neuro-vascularized bone regeneration. Specifically, we incorporated magnesium-ion-modified black phosphorus (BP@Mg) nanosheets into gelatin methacryloyl (GelMA) hydrogel to prepare the upper hydrogel, whereas the bottom hydrogel was designed as a double-network hydrogel system, consisting of two interpenetrating polymer networks composed of GelMA, PEGDA, and ß-TCP nanocrystals. The magnesium ion modification process was developed to enhance BP nanosheet stability and provide a sustained release platform for bioactive ions. Our results demonstrated that the upper layer of hydrogel provided a bionic periosteal structure, which significantly facilitated angiogenesis via induction of endothelial cell migration and presented multiple advantages for the upregulation of nerve-related protein expression in neural stem cells (NSCs). Moreover, the bottom layer of the hydrogel significantly promoted bone marrow mesenchymal stem cells (BMSCs) activity and osteogenic differentiation. We next employed the bilayer hydrogel structure to correct rat skull defects. Based on our radiological and histological examinations, the bilayer hydrogel scaffolds markedly enhanced early vascularization and neurogenesis, which prompted eventual bone regeneration and remodeling. Our current strategy paves way for designing nerve-vascular network biomaterials for bone regeneration.

Silicon-Phosphorus-Nanosheets-Integrated 3D-Printable Hydrogel as a Bioactive and Biodegradable Scaffold for Vascularized Bone Regeneration.

Natural bone is a highly vascularized tissue that relies on the vasculature for blood and nutrients supply to maintain skeletal integrity. Bioactive nanomaterials with the capability of improving vascularized bone regeneration are highly demanded for bone tissue engineering. In this work, 2D silicon phosphorus (SiP) is explored as a new kind of bioactive and biodegradable nanomaterial with excellent angiogenesis and osteogenesis, and a 3D printed biohybrid hydrogel of GelMA-PEGDA incorporated with photocrosslinkable SiP-nanosheet (GelMA-PEGDA/SiPAC) is developed to apply on bone tissue engineering. Findings show that the GelMA-PEGDA/SiPAC possessess excellent biocompatibility and biodegradability, and can sustainably release Si and P elements. Compared with the biohybrid hydrogel scaffolds incorporated with black phosphorus nanosheets, the GelMA-PEGDA/SiPAC can further enhance the osteogenesis of mesenchymal stem cells, and tubular networking of human umbilical vascular endothelial cells. In a rat calvarial bone defect model, the superior angiogenesis and osteogenesis induced by GelMA-PEGDA/SiPAC have been confirmed in vivo. The current strategy paves a new way to design a multifunctional SiP nanocomposite scaffold on mediating the osteogenesis and angiogenesis in one system, and provides a bioactive and biodegradable alternative nanomaterial for tissue engineering and regenerative medicine.

Diagnosis and follow-up MRI evaluation of tennis leg:New understanding of the pathogenesis and imaging.

OBJECTIVES: To evaluate the Magnetic resonance imaging (MRI) findings of patients with a clinical diagnosis of tennis leg and to explore the pathogenesis of tennis leg. METHODS: A retrospective review of 58 (45 men, 13 women; age range, 7-81 years; mean age, 46.7 years) patients with a clinical diagnosis of tennis leg at our hospital during a 64-month period (May 2014 through Sep 2019) was conducted. All patients underwent MRI scan. Follow-up MRI was performed on 4 patients. Images findings, including integrity of the myotendinous junction and tendon of the gastrocnemius and soleus, and presence of fluid collection were analyzed. RESULTS: MRI revealed fluid collection between the medial head of the gastrocnemius and soleus in 44 cases (72.1%). In 25 cases (41.0%), the collected fluid spread to around the medial border of fascia cruris. Fifty-five cases (90.2%) had edema or disruption of the gastrocnemius, with most cases (n = 55) showing edema or disruption of the medial head of the gastrocnemius at the myotendinous junction. Twenty-two (36.1%) cases had edema or disruption of the soleus, with most cases (n = 17) showing edema or disruption of the soleus at the myotendinous junction. Plantaris tendon disruption was observed in 7 cases (11.5%). A thick area of reparative tissue at the distal myotendinous junction of the medial head of the gastrocnemius was observed in all 4 MRI patients followed up. CONCLUSION: Abnormalities of the medial head of the gastrocnemius at the myotendinous junction and tendon appear to be more common than those of the plantaris tendon. Reparative tissue at the distal myotendinous junction of the medial head of the gastrocnemius may be an important specific indication of chronic tennis leg injury.

Adipocyte-secreted microvesicle-derived miR-148a regulates adipogenic and osteogenic differentiation by targeting Wnt5a/Ror2 pathway.

AIMS: Adipocyte-secreted microvesicles (MVs)-derived microRNAs (miRNAs) are relevant to adipogenic and osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) in osteonecrosis of the femoral head (ONFH). Our aims are to investigate the mechanism of adipocyte-derived MVs-miR-148a in ONFH. MATERIALS AND METHODS: Adipocyte-derived MVs were identified via transmission electron microscopy and specific markers expression. The adipogenic and osteogenic differentiation were investigated by Oil-Red O staining, alkaline phosphatase (ALP) activity, Alizarin Red S (ARS) staining and osteogenic or adipogenic factors levels. Genes and proteins expression were detected by using quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting. The relationship between miR-148a and Wnt5a was tested via dual-luciferase reporter analysis. The adipogenic differentiation and osteogenic differentiation in methylprednisolone (MPS)-induced ONFH rat model were assessed via hematoxylin-eosin (HE) staining, and immunohistochemical staining of collagen I (COL I). KEY FINDINGS: Adipocyte-derived MVs promoted adipogenic differentiation via increasing Oil-Red O staining positive cells, adiponectin (Adipoq), acid-binding protein 2 (aP2) and peroxisome proliferator-activated receptor γ (PPAR-γ) levels, and repressed osteogenic differentiation of BMSCs via decreasing ARS staining positive cells, ALP, Runt-related transcription factor 2 (RUNX2) and osteocalcin (OCN) levels. MiR-148a was present in adipocyte-derived MVs, and miR-148a knockdown inhibited adipogenic differentiation and promoted osteogenic differentiation. Furthermore, Wnt5a expression was regulated by miR-148a. MiR-148a overexpression facilitated adipogenic differentiation and suppressed osteogenic differentiation via regulating the Wnt5a/Ror2 pathway. Adipocyte-derived MVs promoted adipogenic differentiation and inhibited osteogenic differentiation in MPS-induced ONFH rat model. SIGNIFICANCE: Adipocyte-derived MVs-miR-148a promoted adipogenic differentiation and suppressed osteogenic differentiation via targeting the Wnt5a/Ror2 pathway.

Platelet-Rich Plasma-Derived Exosomal USP15 Promotes Cutaneous Wound Healing via Deubiquitinating EIF4A1.

Epithelial regeneration is an essential wound healing process, and recent work suggests that different types of exosomes (Exos) can improve wound repair outcomes by promoting such epithelial regeneration. Platelet-rich plasma (PRP) is known to facilitate enhanced wound healing, yet the mechanisms underlying its activity are poorly understood. To explore these mechanisms, we first isolated PRP-derived Exos (PRP-Exos). Using immortalized keratinocytes (HaCaT cells) treated with PBS, PRP, or PRP-Exos, we conducted a series of in vitro Cell Counting Kit-8 (CCK-8), EdU, scratch wound, and transwell assays. We then established a wound defect model in vivo in mice and assessed differences in the mRNA expression within these wounds to better understand the basis for PRP-mediated wound healing. The functions of PRP-Exos and USP15 in the context of wound healing were then confirmed through additional in vitro and in vivo experiments. We found that PRP-Exos effectively promoted the in vitro proliferation, migration, and wound healing activity of HaCaT cells. USP15 was further identified as a key mediator through which these PRP-Exos were able to promote tissue repair both in vitro and in vivo. At a mechanistic level, USP15 enhanced the functional properties of HaCaT cells by promoting EIF4A1 deubiquitination. Thus, PRP-Exos and USP15 represent promising tools that can promote wound healing via enhancing epithelial regeneration.

Mobilization efficiency of granulocyte colony-stimulating factor and stem cell factor to bone marrow mononuclear cells and mechanisms.

The mobilization efficiency of granulocyte colony-stimulating factor (G-CSF) and stem cell factor (SCF) to bone marrow mononuclear cells (MNCs) in mice was observed, and the changes of CXCL12/CXCR4 signal were detected in order to find out the mobilization mechanism of stem cells. Kunming mice were randomly divided into two groups. The mice in treatment group were subjected to subcutaneous injection of G-CSF at a dose of 100 microg/kg and SCF at a dose of 25 microg/kg every day for 5 days, and those in control group were given isodose physiological saline. The MNCs were separated, counted and cultured, and the colony-forming unit-fibroblast (CFU-F) was evaluated. CD34+CXCR4+ MNCs were sorted by flow cytometry. The expression of CXCL12 protein in bone marrow extracellular fluid was detected by ELISA, and that of CXCL12 mRNA in bone marrow was measured by RT-PCR. The results showed that the counts of MNCs in peripheral blood and bone marrow were increased after administration of G-CSF/SCF (P<0.01). The factors had a dramatic effect on the expansion capability of CFU-F (P<0.05). Flow cytometric of bone marrow MNCs surface markers revealed that CD34+CXCR4+ cells accounted for 44.6%+/-8.7% of the total CD34+ MNCs. Moreover, G-CSF/SCF treatment induced a decrease in bone marrow CXCL12 mRNA that closely mirrored the fall in CXCL12 protein. In this study, it is evidenced that G-CSF/SCF can effectively induce MNCs mobilization by disrupting the balance of CXCL12/CXCR4 signaling pathway in the bone marrow and down-regulating the interaction of CXCL12/CXCR4.

LHPP inhibits hepatocellular carcinoma cell growth and metastasis.

Hepatocellular carcinoma (HCC) has a poor prognosis, owing to its high potential for growth and metastasis. In this study, we aimed to investigate the roles of Phospholysine Phosphohistidine Inorganic Pyrophosphate Phosphatase (LHPP)in human HCCcell growth and metastasis. We analyzed the LHPP expression level in human HCC tissues paired normal tissues in the Oncomine database, and assessed the relationship between the LHPP expression levels with HCC patient's overall survival and the prognostic value of LHPP in human HCC by Kaplan-Meier survival analysis. Real-time PCR and Western Blot were used to examine the expression levels of LHPP in normal liver cell line (LO2) and human HCC cell lines (SMCC-7721, HepG2, Huh7, MHCC-97 H, and LM3). Through lentivirus infection, we established human HCC stable cell lines (Huh7 and LM3) overexpressing LHPP. Then, we detected these cell viability, colony , and invasion. Subsequently, we performed the gene set enrichment analysis (GSEA) for the RNA-seq data of HCC patients from TCGA. Finally, we examined the expression level of several oncogenes, including CCNB1, PKM2, MMP7, and MMP9, in these cells via real-time PCR assay. Here, we found thatLHPPis significantly downregulated in the human HCC tissues paired normal tissues. Furthermore, the high expression level of LHPP is associated with better clinical outcomes in human HCC. Overexpression of LHPPinhibitscell growth and metastasis in human HCC cells, and LHPP expression levels negatively correlate with cell cycle and metastasis in HCC tissues. Moreover, the level of LHPP is negatively correlated with CCNB1, PKM2, MMP7, and MMP9 in human HCC cells and HCC tissues. These findings highlight a novel tumor suppressor in human HCC growth and metastasis, and provide a promising diagnostic and prognostic factor for humanHCC.

miR-362-5p promotes cell proliferation and cell cycle progression by targeting GAS7 in acute myeloid leukemia.

Recently, miR-362-5p has attracted special interest as a novel prognostic predictor in acute myeloid leukemia (AML). However, its biological function and underlying molecular mechanism in AML remain to be further defined. Herein, we found that a significant increase in miR-362-5p expression was observed in AML patients and cell lines using quantitative real-time PCR. The expression of miR-362-5p was altered in THP-1 and HL-60 cells by transfecting with miR-362-5p mimic or inhibitor. A series of experiments showed that inhibition of miR-362-5p expression significantly suppressed cell proliferation, induced G0/G1 phase arrest and attenuated tumor growth in vivo. On the contrary, ectopic expression of miR-362-5p resulted in enhanced cell proliferation, cell cycle progression and tumor growth. Moreover, growth arrest-specific 7 (GAS7) was confirmed as a direct target gene of miR-362-5p and was negatively modulated by miR-362-5p. GAS7 overexpression imitated the tumor suppressive effect of silenced miR-362-5p on THP-1 cells. Furthermore, miR-362-5p knockdown or GAS7 overexpression obviously down-regulated the expression levels of PCNA, CDK4 and cyclin D1, but up-regulated p21 expression. Collectively, our findings demonstrate that miR-362-5p exerts oncogenic effects in AML by directly targeting GAS7, which might provide a promising therapeutic target for AML.