Porphyromonas gingivalis promotes the progression of oral squamous cell carcinoma by stimulating the release of neutrophil extracellular traps in the tumor immune microenvironment.

BACKGROUND: The aim of this study was to investigate the impact of Porphyromonas gingivalis (P. gingivalis) on the progression of oral squamous cell carcinoma (OSCC) through neutrophil extracellular traps (NETs) in the tumor immune microenvironment. METHODS: The expression of NETs-related markers was identified through immunohistochemistry, immunofluorescence, and Western blotting in different clinical stages of OSCC samples. The relationship between NETs-related markers and clinicopathological characteristics in 180 samples was analyzed using immunohistochemistry data. Furthermore, the ability to predict the prognosis of OSCC patients was determined by ROC curve analysis and survival analysis. The effect of P. gingivalis on the release of NETs was identified through immunofluorescence and immunohistochemistry, both in vitro and in vivo. CAL27 and SCC25 cell lines were subjected to NETs stimulation to elucidate the influence of NETs on various cellular processes, including cell proliferation, migration, invasion, and metastasis in vitro. Furthermore, the impact of NETs on the growth and metastatic potential of OSCC was assessed using in vivo models involving tumor-bearing mice and tumor metastasis mouse models. RESULTS: Immunochemistry analysis revealed a significant correlation between the NETs-related markers and clinical stage, living status as well as TN stage. P. gingivalis has demonstrated its ability to effectively induce the release of NETs both in vivo and in vitro. NETs have the potential to facilitate cell migration, invasion, and colony formation. Moreover, in vivo experiments have demonstrated that NETs play a pivotal role in promoting tumor metastasis. CONCLUSION: High expression of NETs-related markers demonstrates a strong correlation with the progression of OSCC. Inhibition of the NETs release process stimulated by P. gingivalis and targeted NETs could potentially open up a novel avenue in the field of immunotherapy for patients afflicted with OSCC.

Mena as a key enhancer factor of EMT to promote metastasis of human tongue squamous cell carcinoma.

OBJECTIVE: The aim of this study was to investigate the effect of mammalian-enabled (Mena) on tongue squamous cell carcinoma (TSCC) metastasis and its mechanism. MATERIALS AND METHODS: Immunochemistry was performed to investigate the Mena and tumor-related markers expression, and its clinicopathological characteristics in 46 TSCC specimens. TSCC cell SCC9 and Cal27 untransfected or stable transfected with Mena overexpression and small interfering RNA were used to determine the role of Mena in cell proliferation, cell migration, invasion and metastasis, and EMT-related markers in vitro, and the effect of Mena on TSCC growth and metastasis through tumor-bearing and tumor metastasis immunodeficient mice models in vivo. RESULTS: Immunochemistry showed that the expression of Mena was significantly correlated with lymphatic metastasis and TNM stage, E-cadherin, Vimentin, and MMP2. Mena did not affect cell proliferation and colony formation in vitro, and tumor growth in vivo. However, it promoted cell migration and invasion in vitro, and TSCC metastasis in vivo. CONCLUSIONS: Mena expression is associated with lymphatic metastasis and tumor stage and promotes TSCC invasion and metastasis by inducing the EMT process. Thus, Mena may be a biomarker for prognosis and targeted therapy in TSCC patients.

Effect of concentrated growth factor (CGF) on postoperative sequel of completely impacted lower third molar extraction: a randomized controlled clinical study.

BACKGROUND: The surgical extraction of impacted third molars is one of the most common procedures in oral and maxillofacial surgery, which associated with several postoperative complications. The aim of this clinical trial was to estimate the implication of concentrated growth factor (CGF) on postoperative sequelae after the completely impacted lower third molar extraction. MATERIALS AND METHODS: A total of 74 sides of 37 participants who had completely bilateral impacted lower third molars were enrolled in this split-mouth, randomized single­blind, clinical trial. Surgical extraction was undertaken on both sides of the mandible. Randomization was achieved by opaque, sealed envelopes. The postoperative outcomes including wound healing, swelling and pain were clinically assessed at different-time intervals(1st, 3rd and 7th days). A p-value < 0.05 was considered statistically significant. RESULTS: The wound healing index was significantly better in the test sides (P = 0.001). Regarding the facial swelling, the test sides had significantly less values than the control sides, particularly on the 1st (1.01 ± .57 vs. 1.55 ± .56) and 3rd days (1.42 ± 0.8 vs. 2.63 ± 1.2) postoperatively. Nonetheless, the swelling was disappeared within the 7th day in both sides. The pain scores of visual analog scale were no a statistically significant difference between both sides on the 1st day, meanwhile, the pain scores were significantly lower in the test sides compared with the control sides, especially on the 3rd (P = 0.001) and 7th days (P < 0.001) postoperatively. CONCLUSION: The application of CGF following the surgical extraction of lower third molar has accelerated the healing of soft tissues as well as reduced postoperative sequelae such as swelling and pain. Therefore, the CGF could be promoted among clinicians during the lower third molar surgical extraction. TRIAL REGISTRATION: This study was registered with the TCTR identification number TCTR20210325002 on 25/03/2021 at Thai Clinical Trials Register-Medical Research Foundation of Thailand (MRF). Also it was ethically approved from the institutional ethics committee at the Hospital of Stomatology, Xian Jiaotong University, Xian, China (No: 032), and has been conducted in accordance to the guidelines of the declaration of Helsinki. Written informed consent was obtained from all participants in the study.

Positional Changes of Mandibular Canal Before and After Decompression of Cystic Lesions in the Mandible.

PURPOSE: Significant displacement of the mandibular canal (MC), which occurs frequently in extensive mandibular cystic lesion cases, may raise the risk of inferior alveolar neurovascular bundle injury in surgery. The aim of the present study was to measure the association between positional changes of the MC and the direction (in the coronal plane) of bone expansion of cystic lesions in the mandible. PATIENTS AND METHODS: We performed a retrospective study of patients who had undergone decompression and enucleation surgery from January 2014 to December 2018. Based on coronal planes of cone-beam computerized tomography, the centroids of the expanded mandibles were calculated and considered markers for evaluation of the directions of bone expansion. In addition, the changes in the position of the MC before decompression and enucleation were measured and compared. A Cartesian coordinate system was introduced in this study to illustrate the relationship of positional changes between the displacement of the MC and expansion of the mandible in a straightforward manner. Statistical analysis was performed using the paired t test, unpaired t test, one-way analysis of variance or linear regression as appropriate. RESULTS: Thirty-six patients with an average age of 29.8 years (14 men, 22 women) who received treatment of decompression and enucleation for mandibular cystic lesions were included in this study. The MCs were displaced in the direction toward the lower edge of the mandible and opposite to the direction of mandibular expansion. In addition, the MCs were relocated close to their original location by 1.67 ± 1.45 mm (mean ± standard deviation) approximately 1 year after decompression, accounting for 22.66% of the total displacement. CONCLUSIONS: In mandibular cystic lesion cases, the MCs tend to displace opposite to the direction of mandibular expansion and relocate less after decompression.

Retrospective Clinical Study of Maxillary Sagittal Fractures: Predictors of Postoperative Outcome.

PURPOSE: Because of less attention to the sagittal component of maxillary fractures, these fractures are often misdiagnosed or the reduction is missed leading to maxillary transverse discrepancies. Therefore, the purpose of this study was to identify factors associated with good or adverse postoperative outcomes of maxillary sagittal fractures. MATERIALS AND METHODS: This study was a single-center retrospective cohort study. The sample was composed of cases of maxillary sagittal fractures treated at the Department of Oral and Maxillofacial Surgery, Craniomaxillofacial Trauma Unit of Xi'an Jiaotong University (Xi'an, China) from January 2008 through December 2013. The predictor variables were age, gender, occupation, cause of injury, injury severity, treatment timing, treatment method, and quality of fracture reduction. The outcome variable was the postoperative treatment effect index. Descriptive, bivariate, and multivariate statistics were computed. The P value was set to .05. RESULTS: The sample was composed of 40 cases. The male-to-female ratio was 4:1; the most vulnerable age group was 20 to 30 years (30%); laborers (72.5%) were more prone to injury; and the main cause of injury was motor vehicle accident (62.5%). No cases of isolated sagittal fracture were found and most (35%) occurred with other maxillary fractures, including Le Fort fractures. A statistically significant association between treatment timing and quality of fracture reduction and the postoperative treatment effect index (P < .05) was found. CONCLUSION: The results of this study suggest that better results are achieved when fractured bone is treated sooner. Anatomic repositioning of the fractured bone is the important predictor for good postoperative outcomes.

Using machine learning techniques to predict the risk of osteoporosis based on nationwide chronic disease data.

Osteoporosis is a major public health concern that significantly increases the risk of fractures. The aim of this study was to develop a Machine Learning based predictive model to screen individuals at high risk of osteoporosis based on chronic disease data, thus facilitating early detection and personalized management. A total of 10,000 complete patient records of primary healthcare data in the German Disease Analyzer database (IMS HEALTH) were included, of which 1293 diagnosed with osteoporosis and 8707 without the condition. The demographic characteristics and chronic disease data, including age, gender, lipid disorder, cancer, COPD, hypertension, heart failure, CHD, diabetes, chronic kidney disease, and stroke were collected from electronic health records. Ten different machine learning algorithms were employed to construct the predictive mode. The performance of the model was further validated and the relative importance of features in the model was analyzed. Out of the ten machine learning algorithms, the Stacker model based on Logistic Regression, AdaBoost Classifier, and Gradient Boosting Classifier demonstrated superior performance. The Stacker model demonstrated excellent performance through ten-fold cross-validation on the training set and ROC curve analysis on the test set. The confusion matrix, lift curve and calibration curves indicated that the Stacker model had optimal clinical utility. Further analysis on feature importance highlighted age, gender, lipid metabolism disorders, cancer, and COPD as the top five influential variables. In this study, a predictive model for osteoporosis based on chronic disease data was developed using machine learning. The model shows great potential in early detection and risk stratification of osteoporosis, ultimately facilitating personalized prevention and management strategies.

Construction and validation of a machine learning model for the diagnosis of juvenile idiopathic arthritis based on fecal microbiota.

Purpose: Human gut microbiota has been shown to be significantly associated with various inflammatory diseases. Therefore, this study aimed to develop an excellent auxiliary tool for the diagnosis of juvenile idiopathic arthritis (JIA) based on fecal microbial biomarkers. Method: The fecal metagenomic sequencing data associated with JIA were extracted from NCBI, and the sequencing data were transformed into the relative abundance of microorganisms by professional data cleaning (KneadData, Trimmomatic and Bowtie2) and comparison software (Kraken2 and Bracken). After that, the fecal microbes with high abundance were extracted for subsequent analysis. The extracted fecal microbes were further screened by least absolute shrinkage and selection operator (LASSO) regression, and the selected fecal microbe biomarkers were used for model training. In this study, we constructed six different machine learning (ML) models, and then selected the best model for constructing a JIA diagnostic tool by comparing the performance of the models based on a combined consideration of area under receiver operating characteristic curve (AUC), accuracy, specificity, F1 score, calibration curves and clinical decision curves. In addition, to further explain the model, Permutation Importance analysis and Shapley Additive Explanations (SHAP) were performed to understand the contribution of each biomarker in the prediction process. Result: A total of 231 individuals were included in this study, including 203 JIA patients and Non-JIA individuals. In the analysis of diversity at the genus level, the alpha diversity represented by Shannon value was not significantly different between the two groups, while the belt diversity was slightly different. After selection by LASSO regression, 10 fecal microbe biomarkers were selected for model training. By comparing six different models, the XGB model showed the best performance, which average AUC, accuracy and F1 score were 0.976, 0.914 and 0.952, respectively, thus being used to construct the final JIA diagnosis model. Conclusion: A JIA diagnosis model based on XGB algorithm was constructed with excellent performance, which may assist physicians in early detection of JIA patients and improve the prognosis of JIA patients.

Mechanism of carbon nanotube growth in expanded graphite via catalytic pyrolysis reaction using carbores P as a carbon source.

Carbon nanotubes (CNTs) had potential applications in energy conversion and storage devices, and it could be prepared by expanded graphite loaded with catalyst at high temperature, however, the mechanism of carbon nanotube growth in expanded graphite need further confirmation. In this work, carbon nanotubes' in situ growth in expanded graphite (EG) were prepared via catalytic pyrolysis reaction using carbores P as a carbon source and Co(NO3)3•6H2O as a catalyst. The results of X-ray diffraction (XRD), scanning electron microscope (SEM) and energy dispersive X-ray spectroscope (EDS) indicated the carbon nanotubes could generate in, EG with the presence of carbores P as a carbon source and cobalt nitrate as a catalyst. More interestingly, the growth mechanism of carbon nanotubes could be concluded by the results of differential thermal analysis-thermogravimetry-mass spectrometry (DTA-TG-MS) and X-ray photoelectron spectroscopy (XPS) analysis. The pyrolysis products of carbores P were mainly hydrocarbon gas such as CH4 gas, which reacts with Co(NO3)3·6H2O catalyst to reduces CoOx to Co particles, then the carbon form pyrolysis was deposited the on the surface catalyst Co particles and, after continuous solid dissolution and precipitation, carbon nanotubes were at last generated in EG at last.

PLAU and LAMC2 can predict a poor prognosis in patients with HNSCC.

Objectives: Head and neck squamous cell carcinoma (HNSCC) is the most common malignancy of the head and neck. However, the molecular mechanisms governing the development of HNSCC have not been fully elucidated. Materials and Methods: Differentially expressed genes (DEGs) were screened out from The Cancer Genome Atlas (TCGA) and GSE23036 datasets. Weighted gene coexpression network analysis (WGCNA) was used to reveal the correlations among genes and to search for significantly correlated gene modules. The expression levels of genes in HNSCC and normal samples according to antibody-based detected methods was assessed by utilizing the Human Protein Atlas (HPA). The impact of the selected hub genes on the prognosis of HNSCC patients was assessed by analysing immunohistochemistry (IHC) and immunofluorescence (IF) expression levels and clinical data. Results: Twenty-four genes positively correlated with tumour status and 15 genes negatively correlated with tumour status were screened out by WGCNA. PLAU and LAMC2 were associated with a poor prognosis in patients with HNSCC and were finally screened out and verified by GEPIA and HPA database analysis. Immunohistochemistry of samples collected from 175 patients with HNSCC and subsequent statistical analysis also showed that PLAU and LAMC2 were associated with a poor prognosis in patients with HNSCC, and the levels of these two factors were positively correlated. The expression and co-localization of PLAU and LAMC2 in HNSCC tissues were confirmed by double immunofluorescence labeling. Conclusions: There was a positive correlation between PLAU and LAMC2 expression in HNSCC samples, and PLAU and LAMC2 might be independent prognostic biomarkers for HNSCC.

An Automated Method of 3D Facial Soft Tissue Landmark Prediction Based on Object Detection and Deep Learning.

BACKGROUND: Three-dimensional facial soft tissue landmark prediction is an important tool in dentistry, for which several methods have been developed in recent years, including a deep learning algorithm which relies on converting 3D models into 2D maps, which results in the loss of information and precision. METHODS: This study proposes a neural network architecture capable of directly predicting landmarks from a 3D facial soft tissue model. Firstly, the range of each organ is obtained by an object detection network. Secondly, the prediction networks obtain landmarks from the 3D models of different organs. RESULTS: The mean error of this method in local experiments is 2.62±2.39, which is lower than that in other machine learning algorithms or geometric information algorithms. Additionally, over 72% of the mean error of test data falls within ±2.5 mm, and 100% falls within 3 mm. Moreover, this method can predict 32 landmarks, which is higher than any other machine learning-based algorithm. CONCLUSIONS: According to the results, the proposed method can precisely predict a large number of 3D facial soft tissue landmarks, which gives the feasibility of directly using 3D models for prediction.

Alveolar ridge preservation in post-extraction sockets using concentrated growth factors: a split-mouth, randomized, controlled clinical trial.

Aim: The aim of this clinical trial was to assess the impact of autologous concentrated growth factor (CGF) as a socket-filling material and its ridge preservation properties following the lower third molar extraction. Materials and methods: A total of 60 sides of 30 participants who had completely symmetrical bilateral impacted lower third molars were enrolled. The primary outcome variables of the study were bone height and width, bone density, and socket surface area in the coronal section. Cone beam computed tomography images were obtained immediately after surgery and three months after surgery as a temporal measure. Follow-up data were compared to the baseline using paired and unpaired t-tests. Results: CGF sites had higher values in height and width when compared to control sites (Buccal wall 32.9 ± 3.5 vs 29.4 ± 4.3 mm, Lingual wall 25.4 ± 3.5 vs 23.1 ± 4 mm, and Alveolar bone width 21.07 ± 1.55vs19.53 ± 1.90 mm, respectively). Bone density showed significantly higher values in CGF sites than in control sites (Coronal half 200 ± 127.3 vs -84.1 ± 121.3 and Apical half 406.5 ± 103 vs 64.2 ± 158.6, respectively). There was a significant difference between both sites in the reduction of the periodontal pockets. Conclusion: CGF application following surgical extraction provides an easy, low-cost, and efficient option for alveolar ridge preservation. Thus, the use of CGF by dentists during dental extractions may be encouraged, particularly when alveolar ridge preservation is required. Clinical trial registration: TCTR identification, TCTR20221028003.

Proteomic analysis of mitochondria from infantile hemangioma endothelial cells treated with sodium morrhuate and its liposomal formulation.

Hemangioma is the most common benign tumor of infancy. The aim of this study is to evaluate the biological effects of sodium morrhuate (SM) and its liposomal formulation on infantile hemangioma endothelial cells (IHECs). Morphological analysis revealed that exposure to liposomal sodium morrhuate (LSM) preferentially caused apoptotic death in IHECs, manifested as shrunken configuration and formation of apoptotic bodies. In contrast, necrotic death was prominent in IHECs treated with an equal concentration of SM. By means of proteomic analysis and confirmation experiments, we revealed that the apoptosis-inducing effects of LSM were associated with an upregulation of a set of genes involved in mitochondrial death pathway, including apoptosis-inducing factor, cytochrome c1, caspase-8, and lamin B1. In conclusion, our data highlight the proapoptotic activity of LSM in IHECs through the mitochondrial apoptotic pathway and may provide a promising avenue to treat hemangiomas of infancy.

Clinical Outcomes of Treatment Strategies for Postoperative Plate Fracture and In Situ Fracture of the Femoral Shaft.

Objective: To investigate the treatment and clinical efficacy of postoperative plate fracture and in situ fracture of the femoral stem. Methods: We have retrospectively analyzed the clinical data, revised surgery information, and clinical efficacy of patients with postoperative plate fracture of the femoral stem in our hospital. A total of 33 cases were included whose original fractures were located in the upper and cadaveric femur and treated with paralleling intramedullary pins for revision surgery, as well as patients whose original fractures were located in the lower femur which were fixed with retrograde intramedullary nailing or anatomical locking and compression splints in the distal femur. For the selection of bone grafting, the original fracture site with Fernadez-Esteve scab grades I and II was treated with an autologous iliac bone graft. Postoperatively, patients were evaluated for fracture healing time, the clinical outcome of the affected limb, and complications in the iliac bone donor area. Results: All patients were followed up until fracture healing, and all patients achieved clinical healing with a healing rate of 100% and a mean healing time of 6.3 months. No internal fixation failure such as rebreakage or loosening of the internal fixation occurred in all patients during the follow-up period. According to the Tohner-Wrnch criteria, 23 cases were excellent, 10 cases were good, and 0 cases were poor, with an excellent rate of 100%. Complications in the autologous iliac bone donor area amounted to 36.7%. Conclusion: For patients with original fractures located in the upper femoral segment or cadre, it is recommended to perform revision surgery with a paralleling intramedullary pin, while patients with original fractures located in the lower femoral segment are fixed with the retrograde intramedullary nailing or an anatomical type of distal femoral locking and compression splint. Patients with postoperative plate fractures of the femoral stem do not require routine autologous bone grafting for revision surgery.

Overexpression of Mena is associated with tumor progression and poor prognosis in oral squamous cell carcinoma via EMT.

Background: Mena, a cytoskeletal regulatory protein, is involved in actin-based regulation of cell motility and adhesion, and contributes to tumor invasion and metastasis. However, the role of Mena in oral squamous cell carcinoma remains unclear. This is the first research focusing on the prognostic value of Mena in OSCC. In this study, we aimed to investigate the correlation between Mena expression and clinicopathological significance, as well as prognostic value in OSCC. Methods: Mena gene expression profiles of OSCC and normal tissues were collected from Oncomine, TCGA, and GEO databases. Biological function was analyzed through GO, KEGG and GSEA enrichment. Further, the expression level of Mena and tumor-related markers in 151 OSCC specimens was examined by IHC staining based on tissue microarray. Kaplan-Meier analysis was used to assess the prognostic performance of Mena in OSCC. Result: Mena was generally upregulation in various malignancies, especially OSCC. The functional analyses indicated that Mena was involved in the assembly and regulation of actin, cell movement, and EMT. IHC staining revealed that high expression of Mena in OSCC was correlated with Lymphatic metastasis, TNM stage, E-cadherin, Vimentin, and MMP-2, but insignificantly Ki67. Kaplan-Meier analysis demonstrated that elevated expression of Mena was significantly associated with poor overall survival and disease-free survival of OSCC patients. Conclusion: Mena could be a novel biomarker for predicting the prognosis of OSCC patients, which supports a theoretical basis for developing molecular target therapy.

Cartilage tissue engineering with demineralized bone matrix gelatin and fibrin glue hybrid scaffold: an in vitro study.

To develop a cartilage-like tissue with hybrid scaffolds of demineralized bone matrix gelatin (BMG) and fibrin, rabbit chondrocytes were cultured on hybrid fibrin/BMG scaffolds in vitro. BMG scaffolds were carefully soaked in a chondrocyte-fibrin suspension, which was polymerized by submerging the constructs into thrombin-calcium chloride solution. Engineered cartilage-like tissue grown on the scaffolds was characterized by histology, immunolocalization, scanning electron microscopy, biochemical assays, and analysis of gene expression at different time points of the in vitro culture. The presence of proteoglycan in the fibrin/BMG hybrid constructs was confirmed by positive toluidine blue and alcian blue staining. Collagen type II exhibited intense immunopositivity at the pericellular matrices. Chondrogenic properties were further demonstrated by the expression of gene-encoded cartilage-specific markers, collagen type II, and aggrecan core protein. The glycosaminoglycan production and hydroxyproline content of tissue grown on the fibrin/BMG hybrid scaffolds were higher than that of the BMG group. In conclusion, the fibrin/BMG hybrid scaffolds may serve as a potential cell delivery vehicle and a structural basis for cartilage tissue engineering.

Aggrecanases gene inhibition in chondrocytes: a new possible strategy to relieve immune rejection of transplants.

Cartilage damaged by trauma or degenerative disease has limited intrinsic potential for repair, due to lack of blood supply. The repair and reconstruction of cartilage defects are severe problems, and many patients are eager to find avenues to these matters. Until now, the number of methods used to repair cartilage defects has increased, but all of these have their own advantages and inconveniences, and do not seem to have been optimized. As the source of autologous cartilage is limited and has a high potential donor site morbidity, it is common practice to transplant allogenic cartilage instead. However, immunological rejection will happen accompanied with allogenic cartilage transplantation, affect the long viability of cartilage and result in the absorption of cartilage. Cartilage is an avascular tissue and its extracellular matrix prevents immunization of the host. The extracellular matrix acts as immunological barrier and makes the cartilage be a poor antigen tissue. So it is important to maintain the stability of cartilage matrix. The main features are the loss of aggrecan after cartilage transplantation surgery and aggrecanases play an important role in the cartilage degradation of aggrecan. We hypothesize that if we inhibit the aggrecanases gene of chondrocytes, make the extracellular matrix aggrecan of chondrocytes increasing and immunological rejection problems will be relieved. Accordingly, this will provide a new method for allogenic and tissue engineering cartilage transplantation and cartilage transplantation will be utilized widely for any clinical treatments.

[Application of 3D printing in the treatment of unilateral comminuted zygomatic bone fracture].

To explore the validity of 3D printing technique in the treatment of unilateral comminuted zygomatic bone fracture. METHODS: Twenty-one patients with unilateral comminuted zygomatic bone fracture were included in the present study, which were treated from hospital January 2014 to April 2017. All patients underwent CT scan and the data were imported in Mimics 10.01 software. The zygomatic bone of healthy side was mirrored to the fracture side to rebuild a "perfect" reduction model. Bone fixation plates were pre-modeled on the model printed by a 3D printing machine and used for bone reduction and fixation during operation. Three dimensional measurements were performed to evaluate the validity of 3D printing based on pre- and post-operative three dimensional CT model. SPSS25.0 software package was used to perform paired t test on the measured data. RESULTS: No significant difference were observed between postoperative CT model and preoperative "perfect" reduction model. All patients were satisfied with their facial appearance. CONCLUSIONS: 3D printing technique is helpful to improve the accuracy of reduction of unilateral comminuted zygomatic bone fracture via preoperative pre-modeling.

Effects of RNAi-mediated inhibition of aggrecanase-1 and aggrecanase-2 on rat costochondral chondrocytes in vitro.

AIM: Failure of transplanted cartilage or allogenic chondrocytes is attributed mainly to immunological rejection and cartilage degradation. A major feature is the loss of aggrecan from the cartilage matrix, primarily due to the action of the specific proteinases aggrecanase-1 and aggrecanase-2. The aim of this in vitro study was to determine whether the specific inhibition of aggrecanase-1 and aggrecanase-2 by RNAi would mitigate aggrecan loss from cultured chondrocytes. METHODS: Expression plasmid vectors of shRNA targeting aggrecanase-1 and aggrecanase-2 were constructed and transfected into cultured rattus costochondral chondrocytes. The transfected cells were induced with interleukin-1beta (IL-1beta). Gene mRNA levels were analyzed by RT-PCR. Aggrecan and collagen II content were measured by immunohistochemistry and Western blotting. RESULTS: As the chondrocytes underwent dedifferentiation, aggrecanase-1 increased significantly. The specific inhibition of aggrecanase-1 and aggrecanase-2 by RNAi had no negative effect on the morphology and growth velocity of the chondrocytes. The mRNA of aggrecanase-1 and aggrecanase-2 decreased significantly. The alpha-2-macroglobulin expression level was increased by the shRNA specific for aggrecanase-1. Other genes of the chondrocytic extracellular matrix were not affected. RNAi significantly increased the aggrecan and collagen II content of chondrocytes treated with IL-1beta. CONCLUSION: The results suggest that inhibition of aggrecanase-1 and aggrecanase-2 by RNAi can mitigate aggrecan degradation, without interfering with chondrocytic gene phenotype recovery. RNAi technology can be a useful tool for studying degenerative processes in cartilage.

Propranolol induces hemangioma endothelial cell apoptosis via a p53­BAX mediated pathway.

The use of propranolol for the treatment of infantile hemangioma (IH) has been widely investigated in recent years. However, the underlying therapeutic mechanism of propranolol for the treatment of IH remains poorly understood. The aim of the present study was to investigate the expression of proteins regulated by cellular tumor antigen p53 (p53) in associated apoptosis pathways in IH endothelial cells (HemECs) treated with propranolol. Furthermore, the present study aimed to investigate the exact apoptotic pathway underlying the therapeutic effect of propranolol against IH. In the present study, HemECs were subcultured and investigated using an inverted phase contrast microscope, immunocytochemical staining and a scanning electron microscope (SEM). Experimental groups and blank control groups were prepared. All groups were subjected to drug treatment. A high p53 expression model of HemECs was successfully established via transfection, and a low p53 expression model of HemECs was established using pifithrin­α. The apoptosis rate of each group was determined using Annexin V­fluorescein isothiocyanate/propidium iodide double staining and flow cytometry. The expression levels of downstream proteins regulated by p53 [tumour necrosis factor receptor superfamily member 6 (FAS), p53­induced death domain­containing protein (PIDD), death receptor 5 (DR5), BH3­interacting domain death agonist (BID), apoptosis regulator BAX (BAX), p53 unregulated modulator of apoptosis (PUMA), phosphatidylinositol­glycan biosynthesis class S protein (PIGS), and insulin­like growth factor­binding protein 3 (IGF­BP3)] were revealed in the experimental and control groups via western blotting. Microscopic observation revealed the growth of an adherent monolayer of cells, which were closely packed and exhibited contact inhibition. Immunocytochemical staining demonstrated increased expression of clotting factor VIII. SEM analysis revealed presence of Weibel­Palade bodies. The results of the analyses verified that the cultured cells were HemECs. The staining of the samples resulted in a significantly increased rate of apoptosis in experimental groups compared with the blank control group. This result suggested that there is an association between p53 expression and the rate of apoptosis of propranolol­treated HemECs. The results of the western blot analysis demonstrated an upregulation of BAX expression and a downregulation of IGF­BP3 expression in the HemECs treated with propranolol. There were no significant differences in the expression levels of FAS, DR5, PIDD, BID, PUMA and PIGS between experimental and control groups. This result suggests that p53 has an important role in HemEC apoptosis. The results of the present study additionally suggest that the propranolol­induced HemEC apoptosis pathway is a mitochondrial apoptosis pathway and is regulated by p53­BAX signaling.