Association between circulating cell-free mitochondrial DNA and inflammation factors in noninfectious diseases: A systematic review.

OBJECTIVE: This study aimed to assess the correlation between the circulating cell-free mitochondria DNA and inflammation factors in noninfectious disease by meta-analysis of data from eligible studies. MATERIALS AND METHODS: Through a comprehensive searching of pubmed, embase, web of science, cochrane from establishment of the database to October 31, 2022, studies were selected that investigated the association of circulating cell free mitochondria DNA with inflammatory factors in non-infectious diseases. Studies that met the inclusion criteria and were published in English or Chinese were included. Data of each correlation coefficients were extracted from the paper and 95% confidence intervals were calculated. Sensitivity and heterogeneity tests were carried out for each data. RESULTS: A total of 660 articles were retrieved and 22 were included in this meta-analysis, including 2600 patients. A fixed effects model was employed to examine ISS and IL-8, others were analyzed using random effects models. The correlation coefficient between mtDNA and ISS score was 0.37 (95%CI = [0.232;0.494]), p<0.0001, heterogeneity I2 = 46%, p = 0.11). The correlation coefficients between mtDNA and inflammatory factors are as follows: TNFα, 0.405 [(95%CI = [0.253;0.538], p<0.0001, heterogeneity I2 = 77%, p = 0.0001]. IL-6, 0.469 [(95%CI = [0.296;0.612]), p = 0.0001, heterogeneity I2 = 93%, p<0.0001]. CRP, 0.333[(95%CI = [0.149;0.494]), p = 0.005, heterogeneity I2 = 85%, p<0.0001]. IL-8, 0.343[(95%CI = [0.233;0.524]), p = 0.001, heterogeneity I2 = 50%, p = 0.09]. PCT, 0.333 [(95%CI = [0.06;0.64]), p = 0.09,heterogeneity I2 = 64%,p = 0.06]. There were no significant publication bias for TNFα, IL-6 and CRP. CONSLUSION: Circulating cell free mtDNA was moderate positively correlated with the expression of inflammatory factors and the degree of trauma.

Monitoring angiogenesis in skin autografts using photoacoustic microscopy.

BACKGROUND: Skin autografts have been broadly used to manage the skin and soft tissue defects. It is important for surgeons to assess the vitality of skin autografts via observing the angiogenesis. However, there is lack of reliable approach for giving the quantitative angiogenesis information on the skin autografts. Recently, photoacoustic microscopy imaging has attracted much attention based on its good performance in angiography. METHODS: In this study, we aim to monitor angiogenesis in skin autografts via PAM, and further verify its clinical potential for the early prediction of skin autografts clinical outcome. RESULTS AND CONCLUSIONS: The results indicate that PAM is a feasible, precise, high-resolution, noninvasive technique for the early prediction of necrosis of skin autografts via monitoring the angiogenesis, providing a promising tool for surgeons to use this surgical technology.

NIR-triggered photodynamic therapy of traumatic heterotopic ossification with a type II collagen-targeted photosensitizer.

Traumatic heterotopic ossification (HO) represents an intractable sequela following trauma with no currently effective prophylaxis or treatment. Photodynamic therapy (PDT) is a non-invasive treatment for various proliferative diseases. However, the specific effects of PDT on HO development remain unclear. In this study, the therapeutic potential of a near-infrared (NIR) probe-WL-808, composed of type II collagen-binding peptide (WYRGRL) and a PDT photosensitizer (IR-808), was evaluated for the innovative HO-targeted PDT approach. In vitro studies indicated that WL-808 could induce chondrocyte apoptosis and inhibit cell viability through ROS generation under NIR excitation. In vivo, the efficacy of WL-808-mediated PDT was tested on the tenotomy HO model mice. WL-808 specifically targeted the type II collagen cartilaginous template of HO, promoting cell apoptosis and enhancing extracellular matrix (ECM) degradation under 808 nm NIR excitation, which inhibited the final ectopic bone formation. Moreover, no obvious toxicity or side effects were detected after treatment with WL-808. Taken together, WL-808-mediated PDT significantly diminished ectopic cartilage and subsequent bone formation, providing a new perspective for HO prophylaxis and treatment.

Sustained notch signaling inhibition with a gamma-secretase inhibitor prevents traumatic heterotopic ossification.

Background: Traumatic heterotopic ossification (THO) is a devastating sequela following traumatic injuries and orthopedic surgeries. To date, the exact molecular mechanism of THO formation is still unclear, which hinders the development of effective treatments. The process of THO formation is believed to recapitulate a series of spatiotemporal cellular and signaling events that occur during skeletal development. The Notch signaling pathway is a critical genetic regulator in embryological bone development and fracture healing. However, few data are available concerning whether Notch signaling regulates THO development and maturation. Methods: We firstly detected the expressions of Notch target genes in both mouse and human THO samples with quantitative RT-PCR and immunohistochemistry (IHC). Then, tissue-resident mesenchymal progenitor cells (TMPCs) were isolated, and the abilities of the proliferation and osteogenic and chondrogenic differentiation of TMPCs were examined under the intervention of the gamma-secretase inhibitor-DAPT at different time points. Finally, DAPT was also administrated in THO mice by burn and Achilles tenotomy injury, and ectopic cartilage and bone formation were monitored by histology and micro-CT. Results: Several Notch target genes were upregulated in both mouse and human THO tissues. Sustained Notch signaling inhibition by DAPT reduced proliferation, osteogenic and chondrogenic differentiation of TMPCs in a time-dependent manner. Moreover, DAPT administration within 3 weeks could inhibit ectopic cartilage and bone formation in a mouse THO model without affecting the total body bone mass. Conclusions: The Notch signaling serves as an important therapeutic target during THO formation. And sustained gamma-secretase inhibition by DAPT has great potential in repressing chondrogenic and osteogenic differentiation of TMPCs, as well as inhibited ectopic cartilage and bone formation in vivo. The translational potential of this article: Sustained Notch inhibition via systemic DAPT (or other similar gamma-secretase inhibitors) administration has promising clinical utility for inhibiting THO formation, providing new insight into THO prophylaxis and treatment.

Comparing two surgical approaches for treating multilevel cervical spondylotic myelopathy: A meta-analysis.

PURPOSE: This meta-analysis aims to evaluate the therapeutic efficacy of anterior versus posterior surgical approaches for multisegment cervical spondylotic myelopathy (MCSM). METHODS: Eligible studies published between the period of January 2001 and April 2022 and comparing the anterior and posterior surgical approaches for treating cervical spondylotic myelopathy were retrieved from the PubMed, Web of Science, Embase, and Cochrane databases. RESULTS: A total of 17 articles were selected based on the inclusion and exclusion criteria. This meta-analysis failed to show any significant difference in the duration of surgery, the hospitalization time, or the improvement in the Japanese Orthopedic Association score between the anterior and posterior approaches. The anterior approach, however, exhibited increased efficacy in the improvement of the neck disability index, reduction in the visual analog scale for cervical pain, and improvement in the cervical curvature compared with the posterior approach. CONCLUSION: Bleeding was also less with the anterior surgical approach. The posterior approach provided a significantly higher range of motion of the cervical spine and showed fewer postoperative complications compared with the anterior approach. While both the surgical approaches have good clinical outcomes and show postoperative neurological function improvement, the meta-analysis shows that both anterior and posterior approaches have certain merits and shortcomings. A meta-analysis of a larger number of randomized controlled trials with longer follow-up can conclusively determine which of the surgical approaches is more beneficial in the treatment of MCSM.

Deep-penetration functionalized cuttlefish ink nanoparticles for combating wound infections with synergetic photothermal-immunologic therapy.

The challenge of wound infections post-surgery and open trauma caused by multidrug-resistant bacteria poses a constant threat to clinical treatment. As a promising antimicrobial treatment, photothermal therapy can effectively resolve the problem of drug resistance in conventional antibiotic antimicrobial therapy. Here, we report a deep-penetration functionalized cuttlefish ink nanoparticle (CINP) for photothermal and immunological therapy of wound infections. CINP is decorated with zwitterionic polymer (ZP, namely sulfobetaine methacrylate-methacrylate copolymer) to form CINP@ZP nanoparticles. Natural CINP is found to not only exhibit photothermal destruction of methicillin-resistant Staphylococcus aureus (MRSA) and Escherichia coli (E. coli), but also trigger macrophages-related innate immunity and enhance their antibacterial functions. The ZP coating on the surface of CINP enables nanoparticles to penetrate into deeply infected wound environment. In addition, CINP@ZP is further integrated into the thermosensitive Pluronic F127 gel (CINP@ZP-F127). After in situ spraying gel, CINP@ZP-F127 is also documented notable antibacterial effects in mice wound models infected with MRSA and E. coli. Collectively, this approach combining of photothermal therapy with immunotherapy can promote delivery efficiency of nanoparticles to the deep foci of infective wounds, and effectively eliminate wound infections.

A targeted activatable NIR-II nanoprobe for positive visualization of anastomotic thrombosis and sensitive identification of fresh fibrinolytic thrombus.

Anastomotic thrombosis prevalently causes anastomosis failure, accompanied with ischemia and necrosis, the early diagnosis of which is restricted by inherent shortcomings of traditional imaging techniques in clinic and lack of appropriate prodromal biomarkers for thrombosis initiation. Herein, a fresh thrombus-specific molecular event, protein disulfide isomerase (PDI) is innovatively chosen as the activating factor, and a thrombosis targeting and PDI-responsive turn-on near infrared II (NIR-II) fluorescence nanoprobe is firstly developed. The supramolecular complex-based nanoprobe IR806-PDA@BSA-CREKA is fabricated by assembling NIR-II emitting cyanine derivative IR806-PDA with bovine serum albumin (BSA), which could ameliorate the stability and pharmacokinetics of the nanoprobe, addressing the contradiction in the balance of brightness and biocompatibility. The NIR-II-off nanoprobe exhibits robust turn-on NIR-II fluorescence upon PDI-specific activation, in vitro and in vivo. Of note, the constructed nanoprobe demonstrates superior photophysical stability, efficient fibrin targeting peptide-derived thrombosis binding and a maximum signal-to-background ratio (SBR) of 9.30 for anastomotic thrombosis in NIR-II fluorescent imaging. In conclusion, the exploited strategy enables positive visualized diagnosis for anastomotic thrombosis and dynamic monitoring for thrombolysis of fresh fibrinolytic thrombus, potentially contributes a novel strategy for guiding the therapeutic selection between thrombolysis and thrombectomy for thrombosis treatment in clinic.

Development of a Facile Lysosome-Targeting Nitric Oxide Fluorescent Probe Based on a Novel Reaction Mechanism.

As a ubiquitous signal molecule in biosystems, nitric oxide (NO) plays an important role in many physiological and pathological processes. Therefore, it is of great significance to detect NO in organisms for the study of related diseases. Currently, a variety of NO fluorescent probes have been developed based on several types of reaction mechanisms. However, due to the inherent disadvantages of these reactions, like potential interference by biologically related species, there is a great need to develop NO probes based on the new reactions. Herein, we report our discovery of the unprecedented reaction between a widely used fluorophore of 4-(dicyanomethylene)-2-methyl-6-(p-(dimethylamino)styryl)-4H-pyran (DCM) and NO under mild conditions with fluorescence changes. By the analysis of the structure of the product, we proved that DCM undergoes a particular nitration process and proposed a mechanism for fluorescence changes due to the interruption of the intramolecular charge transfer (ICT) process of DCM by the nitrated product of DCM-NO2. Based on the understanding of this specific reaction, we then easily constructed our lysosomal-localized NO fluorescent probe LysoNO-DCM by linking DCM and a morpholine group, a lysosomal-targeting functional group. LysoNO-DCM exhibits excellent selectivity, sensitivity, pH stability, and outstanding lysosome localization ability with Pearson's colocalization coefficient of up to 0.92 and is successfully applied to the imaging of exogenous and endogenous NO in cells and zebrafish. Our studies expand design methods for NO fluorescence probes based on the novel reaction mechanism and will benefit the studies of this signaling molecule.

Role of Autologous Fat Transplantation Combined with Negative-Pressure Wound Therapy in Treating Rat Diabetic Wounds.

BACKGROUND: Negative-pressure wound therapy (NPWT) and autologous fat transplantation (AFT) are two clinical modalities for plastic and reconstructive surgery. At present, there are few reports on the combination of these two methods in treating diabetic wounds. This study aimed to explore the effect of this combined therapy on diabetic wound healing. METHODS: Full-thickness dorsal cutaneous wounds in rats with streptozotocin-induced diabetes were treated with either NPWT, AFT, or combined therapy. Rats covered with commercial dressings served as the control group. Macroscopic healing kinetics were examined. The levels of inflammation-related factors, such as interleukin-1ß (IL-1ß), interleukin-6 (IL-6), monocyte chemoattractant protein-1, arginase-1, and inducible nitric oxide synthase (iNOS) and angiogenesis-related factors such as vascular endothelial growth factor, were measured in the wound tissues on days 3, 7, and 14; immunohistochemical staining for arginase-1, iNOS, and CD31 was performed on days 3, 7, and 14. The length of reepithelialization was investigated on day 14. RESULTS: The combined therapy promoted faster wound healing than the other treatments. The expression levels of the proinflammatory factors IL-1ß, IL-6, monocyte chemoattractant protein-1 (MCP-1), and iNOS were reduced, and arginase-1 expression was increased compared with those in the other groups. The expression levels of vascular endothelial growth factor and CD31 in the NPWT-combined-with-AFT group were significantly higher than those in the other groups. Reepithelialization was faster in the NPWT-combined-with-AFT group (by day 14) than in the other groups. CONCLUSION: Combining NPWT and AFT can enhance diabetic wound healing by improving wound inflammation and increasing wound vascularization. CLINICAL RELEVANCE STATEMENT: The authors designed a randomized controlled trial of diabetic rats to confirm that NPWT can enhance the vascularization and improve inflammation of the diabetic wound after the autologous fat transplantation treatment. This article may provide a new idea for treating diabetic wounds.

Early diagnosis of heterotopic ossification with a NIR fluorescent probe by targeting type II collagen.

Heterotopic ossification (HO) is a devastating sequela in which the pathologic extracellular matrix of the cartilage and bone forms abnormally in soft tissues following traumatic injuries or orthopaedic surgeries. Early treatment is essential for inhibiting the progression of HO but is currently limited by the absence of sensitive and specific early diagnosis. Herein, this study exploits the enrichment of type II collagen (Col2a1) in the HO cartilage formation stage towards developing a near-infrared (NIR) probe for early HO diagnosis, namely WL-808 by conjugating a Col2a1-binding peptide (WYRGRL) and a cyanine dye (IR-808). WL-808 exhibits high specificity for targeting the cartilage in vitro and in vivo with no apparent cytotoxicity. The NIR signal of WL-808 can be detected in the HO cartilage lesions as early as 1 week after injury when micro-CT cannot show any ectopic bone formation. Moreover, the probe is rarely distributed in the normal knee articular cartilage in vivo via the intravenous administration method. Taken together, WL-808 demonstrates great potential in early HO diagnosis under NIR imaging, facilitating early HO prophylaxis and treatment in the clinic.

Adiponectin protects skeletal muscle from ischaemia-reperfusion injury in mice through miR-21/PI3K/Akt signalling pathway.

Previous studies have confirmed that adiponectin (APN) plays a protective role in myocardial ischaemia-reperfusion (IR) injury, and the aim of this study was to investigate its effect on skeletal muscle. ELISA was used to detect the levels of Creatinine Kinase (CK), LDH, SOD and MDA in the plasma of the lower limbs of mice, and the levels of IL-6, IL-1ß and TNF-α in the gastrocnemius. Quantitative PCR was used to detect the expression level of miR-21. TUNEL staining was used to detect the apoptosis of the gastrocnemius. The expression levels of apoptosis proteins, autophagy marker proteins and downstream target genes of miR-21 in gastrocnemius were detected by Western Blot. The results of this study revealed that APN levels were significantly reduced in gastrocnemius of IR mice. The oxidative stress, inflammatory response, apoptosis and autophagy induced by IR were significantly ameliorated by APN injection. The above effects of APN may be achieved through miR-21/PI3K signalling pathway, as found by interfering gene expression levels with miRNA antagomir and lentiviral injection. Taken together, our study revealed that APN protects skeletal muscle from IR injury through miR-21 /PI3K/Akt signalling pathway through inhibiting inflammatory response, apoptosis and autophagy.

First clinical applications for the NIR-II imaging with ICG in microsurgery.

In microsurgery, it is always difficult to accurately identify the blood supply with ease, such as vascular anastomosis, digit replantation, skin avulsion reconstruction and flap transplantation. Near-infrared window I (NIR-I, 700-900 nm) imaging has many clinical applications, whereas near-infrared window II (NIR-II, 1,000-1700 nm) imaging has emerged as a highly promising novel optical imaging modality and used in a few clinical fields recently, especially its penetration distance and noninvasive characteristics coincide with the needs of microsurgery. Therefore, a portable NIR-II imaging instrument and the Food and Drug Administration (FDA) approved indocyanine green (ICG) were used to improve the operation efficiency in microsurgery of 39 patients in this study. The anastomotic vessels and the salvaged distal limbs were clearly visualized after intravenous injection of ICG. The technique enabled identification of perforator vessels and estimation of perforator areas prior to the flap obtention and made it easier to monitor the prognosis. Overall, this study highlights the use of the portable NIR- II imaging with ICG as an operative evaluation tool can enhance the safety and accuracy of microsurgery.

Comparing the Efficacy of Anatomical Locked Plate Fixation with Coracoclavicular Ligament Augmentation to Hook Plate Fixation in Treating Distal Clavicle Fractures.

OBJECTIVE: Hook plate fixation is the traditional method for treating distal clavicle fractures. However, in recent years, locked plate applications have emerged as a promising treatment method. This study aimed to compare the short- and mid-term clinical efficacy of anatomical locked plate fixation with coracoclavicular ligament augmentation using anchor nails to that of hook plate fixation in treating distal clavicle fractures. METHODS: This was a retrospective single-center cohort study investigating patients with distal clavicle fractures treated between January 2016 and February 2019 in Zhongnan Hospital of Wuhan University. Fifty-nine eligible patients who underwent either anatomical locked plate fixation with coracoclavicular ligament augmentation using anchor nails (LPF&CLA group; 20 patients) or clavicle hook plate fixation (CHPF group; 39 patients) were included. The visual analog scale (VAS) and Constant-Murley shoulder scores were used to assess shoulder function. In addition, the coracoclavicular distance between the affected and unaffected shoulders (ΔCC distance) was measured to assess the reduction. Patients were followed up at 3 months, 6 months, and 1 year postoperatively. The comparisons between the two groups were made using Student's t-test, chi-square test, or Fisher's exact test, if appropriate. RESULTS: Preoperative VAS scores were similar in both groups. At 3- and 6-month follow-up, the VAS score was significantly higher in the CHPF group than in the LPF&CLA group. In contrast, the Constant-Murley shoulder score was significantly lower in the CHPF group than in the LPF&CLA group. When the hook plates were removed, there was no statistical difference in both VAS (0.2 ± 0.4 in LPF&CLA group vs. 0.5 ± 0.5 in CHPF group, p = 0.05) and Constant-Murley shoulder (96.1 ± 3.1 in LPF&CLA group vs. 93.8 ± 5.2 in CHPF group, p = 0.08) scores at the last follow-up. Postoperatively, the ΔCC distance was 2.37 ± 1.93 mm in the LPF&CLA group and -1.56 ± 1.34 mm in the CHPF group. One year after surgery, ΔCC distance increased to 3.96 ± 1.17 mm in the LPF&CLA group and to -0.89 ± 1.39 mm in the CHPF group. CONCLUSION: For distal clavicle fractures in which the coracoclavicular ligament is disrupted, anatomical locked plate fixation with coracoclavicular ligament augmentation achieved better functional recovery and less pain than hook plate fixation at the 6-month follow-up. However, the hook plate provided better reduction throughout the follow-up period and shoulder pain could be relieved using removal surgery. Therefore, locked plates with coracoclavicular ligament augmentation favors post-surgery pain relief while harvesting similar functional outcomes to hook plate fixation.

Dynamically monitoring lymphatic and vascular systems in physiological and pathological conditions of a swine model via a portable NIR-II imaging system with ICG.

Objective: NIR-II imaging with indocyanine green (ICG) has been clinically used in liver tumor resection. However, few data are available concerning the application of ICG-NIR-II in lymphatic and vascular systems in clinic. To expand the application and promote the clinical translation of this approach, we aimed to investigate the feasibility of ICG-NIR-II imaging for monitoring both lymphatic and vascular systems in physiological and pathological conditions using a swine model and compared it to ICG-NIR-I imaging. Methods: we constructed a portable NIR-II imaging system suitable for large animals. Different simulated clinical scenarios in lymphatic and vascular systems of pigs, including lymphatic drainage, lymphorrhea, lymphatic obstruction, lymphatic reconstruction in flaps, venous thrombus formation and vascular anastomosis were modeled to evaluate the reliability of our NIR-II imaging system and the imaging quality of ICG in the NIR-I/II window. Results: Under different simulated clinical scenarios, our portable NIR-II imaging system showed good reliability for pigs. With the help of the portable imaging system, dynamical visualization of lymph vessels, lymph nodes and blood vessels of pigs in different clinical scenarios could be achieved in NIR-II imaging by using the tail fluorescence of ICG. Moreover, ICG-NIR-II imaging has lower background fluorescence and higher resolution than ICG-NIR-I imaging. Conclusions: We demonstrated the first application of a portable NIR-II imaging system for dynamically monitoring both lymphatic and vascular systems in physiological and pathological conditions using a swine model. Our study indicates that ICG-NIR-II imaging be a promising approach for the diagnosis of malfunctions in lymphatic and vascular systems and the surgical navigation of microsurgery and reconstructive surgery.

Rational Design of a Theranostic Agent Triggered by Endogenous Nitric Oxide in a Cellular Model of Alzheimer's Disease.

Oxidative damage caused by upregulated nitric oxide (NO) plays an important role in the pathogenesis of Alzheimer's disease (AD). Currently, stimulus-triggered theranostic agents have received much attention due to benefits on disease imaging and targeted therapeutic effects. However, the development of a theranostic agent triggered by NO for AD remains unexplored. Herein, through the mechanism analysis of the reaction between a fluorophore of 9,14-diphenyl-9,14-dihydrodibenzo[a,c]phenazine (DPAC) and NO, which we occasionally found and thereafter structure optimization of DPAC, a theranostic agent DPAC-(peg)4-memantine was fabricated. In an AD cellular model, DPAC-(peg)4-memantine exhibits NO sensing ability for AD imaging. Meanwhile, DPAC-(peg)4-memantine shows improved therapeutic by targeted drug release triggered by NO and sustained therapeutic effects owing to the synergetic antioxidative abilities via the anti-AD drug and NO scavenging. This work provides an unprecedented avenue for the studies on not only AD but also other diseases with NO upregulation.

Stemness Subtypes and Scoring System Predict Prognosis and Efficacy of Immunotherapy in Soft Tissue Sarcoma.

Tumor stemness has been reported to play important roles in cancers. However, a comprehensive analysis of tumor stemness remains to be performed to investigate the specific mechanisms and practical values of stemness in soft tissue sarcomas (STS). Here, we applied machine learning to muti-omic data of patients from TCGA-SARC and GSE21050 cohorts to reveal important roles of stemness in STS. We demonstrated limited roles of existing mRNAsi in clinical application. Therefore, based on stemness-related signatures (SRSs), we identified three stemness subtypes with distinct stemness, immune, and metabolic characteristics using consensus clustering. The low-stemness subtype had better prognosis, activated innate and adaptive immunity (e.g., infiltrating B, DC, Th1, CD8+ T, activated NK, gamma delta T cells, and M1 macrophages), more enrichment of metabolic pathways, more sites with higher methylation level, higher gene mutations, CNA burdens, and immunogenicity indicators. Furthermore, the 16 SRS-based stemness prognostic index (SPi) was developed, and we found that low-SPi patients with low stemness had better prognosis and other characteristics similar to those in the low-stemness subtype. Besides, low-stemness subtype and low-SPi patients could benefit from immunotherapy. The predictive value of SPi in immunotherapy was more accurate after the addition of MSI into SPi. MSIlowSPilow patients might be more sensitive to immunotherapy. In conclusion, we highlighted mechanisms and practical values of the stemness in STS. We also recommended the combination of MSI and SPi which is a promising tool to predict prognosis and achieve precise treatments of immunotherapy in STS.

MicroRNA-483-5p inhibits osteogenic differentiation of human bone marrow mesenchymal stem cells by targeting the RPL31-mediated RAS/MEK/ERK signaling pathway.

miR-483-5p has been shown to play a key regulatory role in mediating a variety of biological activities. However, there are only a few studies on how miR-483-5p regulates osteogenic differentiation of human bone marrow mesenchymal stem cells (hBMSCs), hence there is a need to explore the role and mechanism of miR-483-5p in regulating the osteogenic differentiation of hBMSCs. To that end, we used bioinformatics and cell experiments to confirm our hypothesis, a miRNA microarray dataset (GSE74209) and a mRNA dataset (GSE56816) were obtained from the GEO database, and the differentially expressed miRNAs (DE-miRNAs) and mRNAs (DE-mRNAs) were screened. In total, We found that the up-regulated candidate target genes were significantly enriched in the MAPK signaling pathway. Using the MCODE plug-in from Cytoscape, we identified RPL31 as the seed gene in the third major module. And we successfully established a possible miRNA-hub gene regulatory network. More importantly, we confirmed that the expression of miR-483-5p was down-regulated while the expression of RPL31 was up-regulated during osteogenic induction. Overexpression of miR-483-5p significantly inhibited osteogenic differentiation. In addition, western blot was used to measure protein levels of RPL31 and phosphorylated MEK/ERK family members. We demonstrated that miR-483-5p inhibits the RAS/MEK/ERK signaling pathway by targeting RPL31 and inhibiting its expression, thereby playing an inhibitory role in osteogenic differentiation. In light of our findings, RPL31 can be used as a novel therapeutic target for bone defects and osteoporosis, we reveal a newly discovered mechanism of osteogenic differentiation and provide a new strategy for treating osteoporosis and related diseases.

Corticoperiosteal Pedicle Flap of Greater Trochanter for Salvage of Old Femoral Neck Fracture in Children: Outcomes After a Minimum 5 Years Follow-up.

BACKGROUND: Old femoral neck fracture (OFNF) generally refers to fractures for more than 3 weeks. Corticoperiosteal pedicle flap of greater trochanter (CPPF-GT) was designed to restore blood supply and donor bone for OFNF. This study aimed to assess the efficacy and radiographic results of CPPF-GT for treatment of OFNF in children after a minimum 5 years follow-up. METHODS: Twenty-three patients with OFNFs, age from 8 to 16 years old, who underwent open reductions, fracture fixations, and transpositions of CPPF-GTs were retrospectively reviewed. Clinical and radiological outcomes, including union, nonunion, avascular necrosis of femoral head, limb shortening, coxa vara, premature epiphyseal closure and Ratliff's assessment, were investigated in the postoperative follow-up. The results were compared with previously published joint-salvage study of OFNFs. RESULTS: All patients were followed for an average of 5.9 years (range: 5 to 10 years). All cases (100%) achieved hip unions at an average duration of 3.5 months (range: 2.5 to 5 months). No nonunion hip was observed. Three hips (13.0%) progressed to avascular necrosis of femoral head after 1.5 to 3 years of operation, respectively, and the revision surgeries to hip replacements were conducted. Premature epiphyseal closures were observed in 3 hips. Three hips visibly presented an average 2 cm shortening of the femoral neck. Coxa vara deformities were observed in 2 hips. According to the Ratliff's criteria, there were 20 cases (87.0%) with satisfactory union, good results were achieved in 17 cases, fair results in 3 cases, and poor result in 3 cases. CONCLUSIONS: Corticoperiosteal pedicle flap of greater trochanter is an effective and desirable option for treating old femoral neck fractures in children with a low rate of avascular necrosis and without nonunion. LEVEL OF EVIDENCE: Therapeutic Level IV. See Instructions for Authors for a complete description of levels of evidence.

Fibro-Osseous Pseudotumor.

Fibro-osseous pseudotumor is an extremely rare subcutaneous benign ossifying lesion associated with bone formation that is most commonly seen in the hands, followed by the toes. Because the tumor has a certain degree of invasiveness, it is often mistaken for malignancy, which leads to radical, excessive treatment. Our case involved a 32-year-old man with lesions on the left index finger. We documented the detailed data of diagnosis, treatment, and follow-up. We also conducted a review and summarized the published cases to advance our understanding of the disease, provide more accurate diagnostic criteria, and avoid inappropriate surgical procedures. [Orthopedics. 2021;44(6):e713-e718.].

Application and Clinical Effectiveness of Antibiotic-Loaded Bone Cement to Promote Soft Tissue Granulation in the Treatment of Neuropathic Diabetic Foot Ulcers Complicated by Osteomyelitis: A Randomized Controlled Trial.

This study explored the clinical effectiveness of antibiotic-loaded bone cement on primary treatment of diabetic foot infection. This is a randomized controlled study, including thirty-six patients with diabetic foot ulcer complicated by osteomyelitis who had undergone treatment between May 2018 and December 2019. Patients were randomly divided into control group (group A) and study group (group B). Patients in the intervention group received antibiotic-loaded bone cement repair as primary treatment, while patients in the control group received conventional vacuum sealing draining treatment. Clinical endpoints were assessed and compared between the two groups, including wound healing time, wound bacterial conversion, NRS pain score, number of wound dressing changes, and average hospitalization time. All patients were followed up for a period of 12 months after discharge. Results show that compared with the control group, patients in the study group had significant difference in the number of patients for baseline pathogens eradication, short NRS pain score, hospital length of stay and cost, wound surface reduction, healing time, low rate of complications, and infection recurrence. Based on the findings, we conclude that antibiotic-loaded bone cement can be used for treatment of wound in patient with diabetic foot infection. It can help to control wound infections, shorten hospital length of stay, reduce medical cost, and relieve both doctors' and patients' burden. The application of antibiotic-loaded bone cement is suitable for diabetic wound with soft tissue infection or osteomyelitis.