Low-dose celecoxib-loaded PCL fibers reverse intervertebral disc degeneration by up-regulating CHSY3 expression.

Intervertebral disc degeneration (IDD) has been identified as one of the predominant factors leading to persistent low back pain and disability in middle-aged and elderly people. Dysregulation of Prostaglandin E2 (PGE2) can cause IDD, while low-dose celecoxib can maintain PGE2 at the physiological level and activate the skeletal interoception. Here, as nano fibers have been extensively used in the treatment of IDD, novel polycaprolactone (PCL) nano fibers loaded with low-dose celecoxib were fabricated for IDD treatment. In vitro studies demonstrated that the nano fibers had the ability of releasing low-dose celecoxib slowly and sustainably and maintain PGE2. Meanwhile, in a puncture-induced rabbit IDD model, the nano fibers reversed IDD. Furthermore, low-dose celecoxib released from the nano fibers was firstly proved to promote CHSY3 expression. In a lumbar spine instability-induced mouse IDD model, low-dose celecoxib inhibited IDD in CHSY3wt mice rather than CHSY3-/- mice. This model indicated that CHSY3 was indispensable for low-dose celecoxib to alleviate IDD. In conclusion, this study developed a novel low-dose celecoxib-loaded PCL nano fibers to reverse IDD by maintaining PGE2 at the physiological level and promoting CHSY3 expression.

Clinical Features and Influencing Factors for the Prognosis of Patients With Sudden Deafness.

Backgrounds: Studies on risk factors influencing the prognosis of patients with sudden onset deafness are lacking. Methods: From March 2018 to March 2021, 500 patients, from the Tongde Hospital in Zhejiang Province, with sudden onset deafness were enrolled. We collected clinical information from the hospital medical records, including certain demographic characteristics, information related to sudden-onset deafness, and laboratory parameters. Univariate and multivariate analyses were performed to determine independent prognostic risk factors for patients with sudden deafness. Additionally, we also employed orthogonal partial least squares discriminant analysis (OPLS-DA) to analyze the data of these enrolled patients. Results: The baseline clinical characteristics of the enrolled patients were analyzed. Based on their prognoses, the included patients were divided into the overall effective and ineffective groups. Between these two groups, the univariate and multivariate analyses were performed. Age, type of hearing curve at the initial diagnosis, acute phase, and sudden deafness site were found to be independently associated with the prognoses of patients with sudden deafness (all P < 0.05). Through the OPLS-DA, the sudden deafness site was found to be an indicator with the highest predictive power. Conclusions: Age, type of hearing curve at the initial diagnosis, acute phase, and sudden deafness site were all independently correlated with the prognoses of patients with sudden deafness and, therefore, need to be emphasized.

Saxagliptin enhances osteogenic differentiation in MC3T3-E1 cells, dependent on the activation of AMP-activated protein kinase α (AMPKα)/runt-related transcription factor-2 (Runx-2).

Osteoporosis is a metabolic bone disease commonly observed in the elderly, and its pathogenesis is associated with declined osteogenic differentiation. Osteogenic differentiation could be facilitated by the activation of the AMP-activated protein kinase (AMPK) pathway. Saxagliptin, an anti-diabetic agent with inhibitory effects against dipeptidyl peptidase 4 (DPP-4), has been recently reported to induce the activation of the AMPK pathway. The present study proposes to explore the function and mechanism of Saxagliptin in osteogenic differentiation. Osteogenic differentiation induction medium (ODIM) was utilized to induce osteogenic differentiation in MC3T3-E1 cells. Significantly increased mineral nodule formation, elevated alkaline phosphatase (ALP) activity, and upregulated expression of osteogenic marker genes activating transcription factor-4 (ATF-4), osteopontin (OPN), and type I collagen (Col1) were observed in ODIM-cultured MC3T3-E1 cells, all of which were further enhanced by the introduction of Saxagliptin. The elevated expression level of runt-related transcription factor-2 (Runx-2), an important transcriptional factor involved in the progression of osteogenic differentiation, in ODIM-cultured MC3T3-E1 cells was further promoted by Saxagliptin. The AMPK pathway in ODIM-cultured MC3T3-E1 cells was significantly activated by Saxagliptin, and the functions of Saxagliptin in promoting osteogenic differentiation were abolished by compound C, the inhibitor of the AMPK pathway. Conclusively, Saxagliptin enhanced osteogenic differentiation in MC3T3-E1 cells, dependent on the activation of AMPKα/RUNX-2.

MicroRNA-222 alleviates radiation-induced apoptosis by targeting BCL2L11 in cochlea hair cells.

Radiation-induced hair cell injury is detrimental for human health but the underlying mechanism is not clear. MicroRNAs (miRNAs) have critical roles in various types of cellular biological processes. The present study investigated the role of miR-222 in the regulation of ionizing radiation (IR)-induced cell injury in auditory cells and its underlying mechanism. Real-time PCR was performed to identify the expression profile of miR-222 in the cochlea hair cell line HEI-OC1 after IR exposure. miRNA mimics or inhibitor-mediated up- or down-regulation of indicated miRNA was applied to characterize the biological effects of miR-222 using MTT, apoptosis and DNA damage assay. Bioinformatics analyses and luciferase reporter assays were applied to identify an miRNA target gene. Our study confirmed that IR treatment significantly suppressed miR-222 levels in a dose-dependent manner. Up-regulation of miR-222 enhances cell viability and alleviated IR-induced apoptosis and DNA damage in HEI-OC1 cells. In addition, BCL-2-like protein 11 (BCL2L11) was validated as a direct target of miR-222. Overexpression of BCL2L11 abolished the protective effects of miR-222 in IR-treated HEI-OC1 cells. Moreover, miR-222 alleviated IR-induced apoptosis and DNA damage by directly targeting BCL2L11. The present study demonstrates that miR-222 exhibits protective effects against irradiation-induced cell injury by directly targeting BCL2L11 in cochlear cells.

Immune cell infiltration and related core genes expression characteristics in eosinophilic and non-eosinophilic chronic rhinosinusitis with nasal polyps.

Chronic rhinosinusitis with nasal polyps (CRSwNP) refers to chronic inflammation of the sinonasal mucosa. It can either be eosinophilic (ECRSwNP) or non-eosinophilic (non-ECRSwNP). However, immune cell infiltration in the microenvironment and pathogenesis of ECRSwNP and non-ECRSwNP are still unclear. The aim of the present study was to assess the immune cell infiltration and molecular mechanisms of ECRSwNP and non-ECRSwNP. In the present study, 22 immune cell types in ECRSwNP and non-ECRSwNP were investigated by CIBERSORT based on transcriptome data. The core gene related pathophysiology of CRSwNP was analyzed using Weighted Gene Correlation Network Analysis according to the phenotype of the infiltrated eosinophils and nasal polyps (NP). A total of four types of immune cells (mast cells, activated dendritic cells, M2 macrophages and activated natural killer cells) were demonstrated to have a direct and indirect correlation with eosinophilic infiltration in ECRSwNP. M1 macrophages and activated CD4+ memory T cells were correlated with major immune cell types in non-ECRSwNP. NP could affect the expression of 'olfactory receptor activity' and 'channel activity' genes to impair the olfactory signaling pathway and neuroactive ligand receptor pathway. 'Cell adhesion molecule binding', 'cytokine receptor binding' and 'glucocorticoid receptor binding' were significantly enriched in ECRSwNP, whereas epithelial cell injury, autophagy and the mTOR pathway (hsa04140 and hsa04150) may serve an important role in the pathogenesis of non-ECRSwNP. There were significantly different immune cell infiltration and related core genes expression characteristics between ECRSwNP and non-ECRSwNP. The results of the present study provide an improved basis for elucidation of the mechanism and treatment of CRSwNP.

FOXD3 inhibits cell proliferation, migration, and invasion in nasopharyngeal carcinoma through regulation of the PI3K-Akt pathway.

FOXD3 has been found previously to positively regulate miR-26b, a tumor inhibitor of nasopharyngeal carcinoma (NPC). However, FOXD3's precise function and associated mechanism of action in NPC have not yet been investigated. In this study, the expression of FOXD3 mRNA and protein was evaluated using RT-qPCR, western blotting, and immunohistochemistry. Protein levels involved in the phosphoinositide 3-kinase - protein kinase B (PI3K-Akt) pathway were assessed by western blot, and cell proliferation was determined by MTT and colony forming assays. Additionally, cell apoptosis was assessed by flow cytometric assay. Finally, the migration and invasion capabilities of the NPC cells were determined using wound healing and Transwell assays. We found that FOXD3 levels were relatively low in NPC tissue and cells, while an increase caused the inhibition of the PI3K-Akt pathway. Functional experiments found that overexpression of FOXD3 suppressed cell proliferation, migration, and invasion and enhanced cell apoptosis in NPC C6661 cells. IGF-1, an activator of the PI3K-Akt pathway, reversed the inhibitory effect of FOXD3. Furthermore, we found upregulation of the PI3K-Akt pathway and upregulation of the inhibitory effects of FOXD3 on C6661 cellular activities. In conclusion, FOXD3 negatively affected the PI3K-Akt pathway to restrain the processes involved in C6661 cell pathology. These findings further exposed the function and downstream axis of FOXD3 in NPC and displayed a promising new target for NPC therapy.

Long non-coding RNA LINC00460 promotes head and neck squamous cell carcinoma cell progression by sponging miR-612 to up-regulate AKT2.

LncRNAs (long noncoding RNAs) have been shown to be potentially critical regulators in head and neck squamous cell carcinoma (HNSCC). LncRNA LINC00460 (long intergenic non-protein coding RNA 460), an "oncogene", regulates progression of various tumors. However, the tumorigenic mechanism of LINC00460 on HNSCC is yet to be investigated. In the current study, we discovered that LINC00460 was relatively up-regulated in both HNSCC cancer tissues and cell lines, and predicted a poor prognosis in HNSCC patients. Gain- and loss-of functional studies established that over-expression of LINC00460 promoted cell proliferation, invasion and migration of HNSCC cells in vitro, while the promotion abilities were suppressed via knockdown of LINC00460. Our results identified miR-612 as a novel target of LINC00460, whose expression suggested a negative correlation with LINC00460 in HNSCC tissues and cell lines. LINC00460 increased the expression of serine/threonine kinase AKT2 via sponging miR-612. Rescue experiments indicated that LINC00460 could promote HNSCC progression partially through inhibition of miR-612. Subcutaneous xenotransplanted tumor model confirmed that interference of LINC00460 suppressed in vivo tumorigenic ability of HNSCC via down-regulation of AKT2. In conclusion, our findings clarified the biologic significance of LINC00460/miR-612/AKT2 axis in HNSCC progression and provided novel evidence that LINC00460 may be a new potential therapeutic target for HNSCC.

True hypopharyngeal carcinosarcoma: a case report and literature review.

Objective Carcinosarcoma consists of carcinomatous and sarcomatous tissues and is an aggressive malignant tumor. It is rarely reported in the hypopharynx. Methods A 72-year-old man presented with dysphagia and dyspnea. Laryngoscopy, computed tomography (CT), and 18F-fluorodeoxyglucose positron emission tomography/CT (18F-FDG PET/CT) showed a neoplasm on the left posterior hypopharyngeal wall. The patient underwent bilateral neck dissection and excision of the hypopharyngeal cancer followed by postoperative radiation therapy. Results Immunohistochemistry revealed carcinomatous cells with membrane positivity for cytokeratin, glucose transporter-1 (GLUT-1), phosphoinositide-3 kinase (PI3K), hypoxia-inducible factor-1α (HIF-1α), and hexokinase-II as well as sarcomatous cells with membrane positivity for smooth muscle actin, GLUT-1, HIF-1α, and PI3K. Histopathology and immunohistochemistry revealed a true carcinosarcoma of the hypopharynx (pT3N0M0, Stage III). Conclusions Thorough immunohistochemistry is required for a correct diagnosis of hypopharyngeal carcinosarcoma. 18F-FDG PET/CT may help to distinguish hypopharyngeal carcinosarcoma from benign tumors.

Healing improvement after rotator cuff repair using gelatin-grafted poly(L-lactide) electrospun fibrous membranes.

BACKGROUND: Rotator cuff tears (RCTs) are a common cause of shoulder pain and disability in middle and older age. Despite improvements in the understanding of this disease process and advances in surgical treatment, rotator cuff (RC) repair failure rates remain high. Insufficient healing capacity is likely the main factor for failure of reconstruction. MATERIALS AND METHODS: We fabricated implantable biodegradable gelatin-grafted poly(L-lactide) (PLLA) fibrous membranes using electrospinning technology and evaluated them using in vitro cell proliferation assays. Then, we established chronic rat RCT models and randomly assigned rats into one of three groups. In group 1 (n = 48), the detached supraspinatus tendon was repaired to its anatomic footprint (transosseous repair). In groups 2 and 3, the rats underwent transosseous repair and were implanted with either pure PLLA membranes (n = 48) or gelatin-PLLA membranes (n = 48) to augment the repairs. The animals were killed at 2, 4, and 8 wk postoperatively, which was followed by histomorphometric and biomechanical evaluation. RESULTS: Histologic observations revealed that gelatin-PLLA membranes have excellent biocompatibility and biodegradability. At 2, 4, and 8 wk postoperatively, the gelatin-PLLA membranes significantly increased the area of glycosaminoglycan staining at the tendon-bone interface compared with the control group (P < 0.05) and significantly improved collagen organization, as measured by birefringence under polarized light at the healing enthesis compared with the control and PLLA groups (P < 0.05). Biomechanical testing revealed that the gelatin-PLLA group had a greater ultimate load to failure and stiffness than the control group at 4 and 8 wk (P < 0.05). The gelatin-PLLA membranes had the highest stress of the healing enthesis. CONCLUSIONS: Local application of gelatin-PLLA fibrous membranes to the healing tendon-bone interface after RC repair in a rat chronic RCT model was found to strengthen the healing enthesis, increase the area of fibrocartilage, and improve collagen organization compared with repair alone. Augmentation with gelatin-grafted PLLA may enhance healing after RC repair and might eventually lead to improvement of clinical surgical outcomes.

Open reduction and bi-columnar internal fixation of intra-articular distal humerus fractures through a combined medial and lateral approach.

PURPOSE: The aim of this retrospective study was to investigate the suitability of bi-columnar internal fixation through a combined medial and lateral approach for the treatment of intra-articular distal humerus fractures. METHODS: Nineteen cases of intra-articular distal humerus fractures were treated with open reduction and bi-columnar internal fixation through a combined medial and lateral approach. The reduction in the articular surface and functional recovery of the affected elbows was assessed at an average follow-up of 15.8 ± 7.9 (7-43) months. RESULTS: The gap in the main articular fragments was less than 1 mm in 16 cases, while a gap of more than 1 mm and less than 2 mm was identified in 2 cases and of 3.7 mm in one case. All the fractures were united. At the latest follow-up, the mean flexion-extension of the elbows was 113.4° ± 20.7°, while the pronation-supination of the forearms was 158.3° ± 8.5°, and the mean Mayo Elbow Performance Index was 93.7 ± 9.1 points, leading to 13 excellent outcomes, and 6 with good results. CONCLUSIONS: Intra-articular fractures of the distal humerus can be effectively treated by open reduction and internal fixation through a combined medial and lateral approach at the elbow.

Comparison of internal and external fixation of distal radius fractures.

BACKGROUND AND PURPOSE: There is no consensus on the difference in effects of internal fixation (IF) and external fixation (EF) on outcomes for the treatment of distal radius fractures. We performed a meta-analysis of randomized clinical studies. METHODS: We searched the literature and included studies that compared the effects of IF and EF on the treatment of distal radius fractures. Statistically, we pooled patient data using standard meta-analytic methods. For the continuous variables, the weighted mean difference (WMD) was used. For dichotomous data, the relative risk (RR) was calculated. RESULTS: 10 studies were eligible for data extraction. The pooled data showed that compared with EF, IF led to statistically significantly better Disabilities of the Arm, Shoulder, and Hand (DASH) scores at 12 months postoperatively, recovery of forearm supination at 3 months, and restoration of volar tilt and radial inclination. IF using volar locking plates resulted in better DASH scores than EF at 3 and 6 months, but the trend diminished over time; at 12 months postoperatively, the scores were not statistically significant. Compared with EF, IF led to fewer minor surgical complications. INTERPRETATION: For surgical treatment of distal radius fractures, IF yields better functional outcomes, forearm supination, restoration of anatomic volar tilt and radial inclination, and fewer minor complications. The patients who received IF using volar locking plates for the treatment of distal radius recovered more quickly than did patients who received EF.

Platelet-rich plasma enhances autograft revascularization and reinnervation in a dog model of anterior cruciate ligament reconstruction.

BACKGROUND: Autologous platelet-rich plasma (PRP) has been investigated as a potential promoter of tendon healing that affects the anterior cruciate ligament (ACL) graft maturation process. However, the influence of PRP on revascularization and reinnervation during the ACL graft remodeling has never been investigated. MATERIALS AND METHODS: We randomly assigned healthy and mature beagles to one of four groups. In group 1 (PRP group), we treated the ACL grafts with PRP. In group 2 (control group), we treated the ACL grafts with saline. In group 3 (sham group), we exposed only the knee joints. In group 4 (normal control group), no surgery was performed on the knees. We dissected the ligament tissue at 2, 6, and 12 wk after surgery and performed real-time polymerase chain reaction using primers for cluster of differentiation molecule 31, vascular endothelial growth factor, thrombospondin-1 (TSP-1), neurotrophin-3, growth-associated protein-43 (GAP-43), and nerve growth factor. RESULTS: We observed the increased expression of vascular endothelial growth factor, TSP-1, neurotrophin-3, GAP-43, and nerve growth factor mRNA in group 1 at 2, 6, and 12 wk after surgery, compared with that in group 2 (P < 0.05). We also detected increased levels of cluster of differentiation molecule 31 expression in group 1 (P < 0.05) at 2 and 6 wk after surgery. The levels of TSP-1 and GAP-43 mRNA were significantly increased in group 3 compared with those in group 4 at 2 wk after surgery (P < 0.05). CONCLUSIONS: During graft remodeling, we observed a time-dependent change in gene expression after ACL reconstruction surgery. In addition, these results demonstrate that PRP alters the expression of some target genes at certain times, particularly during the early stages of graft remodeling. Platelet-rich plasma could promote revascularization and reinnervation, which might explain the enhancing effect of PRP on ACL graft maturation.

The effect of platelet-rich plasma on patterns of gene expression in a dog model of anterior cruciate ligament reconstruction.

BACKGROUND: Autologous platelet-rich plasma (PRP) has been investigated as a potential promoter of tendon healing and has an enhancing effect on the anterior cruciate ligament (ACL) graft maturation process. However, the influence of PRP on the synthesis and degradation of the extracellular matrix during the ACL graft remodeling process has never been investigated. MATERIALS AND METHODS: Healthy and mature beagle dogs were randomly assigned to one of four groups: in group I (PRP group), ACL grafts were treated with PRP; in group II (control group), ACL grafts were treated with saline; in group III (sham group), only the knee joints were exposed; in group IV (normal control group), no surgery was performed to the knees. Ligament tissue was dissected at 2, 6, and 12 wk after surgery, and real-time PCR was performed using primers for growth factor-ß1 (TGF-ß1), collagen type1A1 (COL1Al), collagen type3A1 (COL3A1), decorin, biglycan, matrix metalloproteinase-1 (MMP-1), matrix metalloproteinase-13 (MMP-13), and tissue inhibitor of metalloproteinase-1 (TIMP-1). RESULT: In group I, the messenger RNA (mRNA) levels of collagen type 1A1, biglycan, and MMP-1 all increased 2, 6, and 12 wk after surgery, compared with group II (P < 0.05). At 2 and 6 wk after surgery, increased levels of COL3A1, MMP-1, and MMP-13 mRNA were also detected in group I (P < 0.05). Increased levels of TGF-ß1 mRNA was observed at 6 and 12 wk in group I after surgery (P < 0.05). CONCLUSIONS: During the graft remodeling process, we observed a time-dependent change of gene expression following ACL reconstruction surgery. Furthermore, our results demonstrate that PRP alters the expression of some target genes at certain time points, especially during the early stages of graft remodeling, which might explain the enhancing effect of PRP on the ACL graft maturation process.

[Relationship between bacterial biofilms and chronic rhinosinusitis by scanning electron microscopic study].

OBJECTIVE: To determine the presence and the morphological features of bacterial biofilms in surgical specimens of patients with chronic rhinosinusitis (CRS) compared with control patients without CRS by scanning electron microscopy (SEM), and to evaluate the role of biofilm on the pathogenesis of CRS. METHODS: Fifteen patients with CRS undergoing endoscopic sinus surgery and 11 control patients with fracture of nasal bone were enrolled in this study. Clinical information was recorded from each patient. All patients underwent a thorough otolaryngological examination, preoperative paranasal sinus computerized tomography (CT) scanning which were evaluated according to the Lund-Mckay CT scoring system. All the samples including uncinate process, ethmoid mucosa from CRS group and uncinate process, ethmoid bulla from control group were prepared using standard methods for SEM. The presence of bacterial biofilms on the samples of two groups was observed by SEM. Statistical analysis was performed using SPSS 13.0. Continuous data was analyzed by Student t test and dichotomous data was analyzed by chi² or Fisher exact test. P was considered to be significant at a level of 0.05. RESULTS: Nine (60.0%) of the 15 patients were found to have evidence of biofilms. In control group, only 1 (9.1%) of 11 patients had biofilm. The difference was statistical significant (chi² = 6.949, P < 0.01). All controls except one had healthy appearing cilia and goblet cells without biofilms. All the 16 CRS patients showed aberrant findings of the mucosal surface with variation in degrees of severity from disarrayed cilia to complete absence of cilia and goblet cells. Among them the typical morphologic feature such as water channels, 3-D structure, and matrix-embedding spherical or elliptical bodies were noted in 9 cases. Five samples including one case from control showed cilia aggregation. The preoperative CT scores of the CRS patients with biofilms (n = 9) were significantly higher than those without biofilms (n = 6, t = 2.14, P < 0.05). CONCLUSIONS: The typical morphologic feature of BF such as water channels, 3-D structure, and matrix-embedding spherical or elliptical bodies were noted in sinus mucosa of patients with CRS by the SEM. The positive rate of bacterial biofilms in CRS group was significantly higher compared to control group, which indicated bacterial biofilms might play an important role in the pathogenesis of CRS. Besides the typical bacterial biofilm features, cilia aggregation was found in five cases of CRS patients. We consider cilia aggregation can be regarded as one morphologic feature of bacterial biofilm in nasal mucosa, which needs further study. The presence of bacterial biofilms in CRS patients is associated with paranasal CT scores, which indicates that bacterial biofilm is correlated with the severity of CRS.