Decoding Algorithm of Motor Imagery Electroencephalogram Signal Based on CLRNet Network Model.

EEG decoding based on motor imagery is an important part of brain-computer interface technology and is an important indicator that determines the overall performance of the brain-computer interface. Due to the complexity of motor imagery EEG feature analysis, traditional classification models rely heavily on the signal preprocessing and feature design stages. End-to-end neural networks in deep learning have been applied to the classification task processing of motor imagery EEG and have shown good results. This study uses a combination of a convolutional neural network (CNN) and a long short-term memory (LSTM) network to obtain spatial information and temporal correlation from EEG signals. The use of cross-layer connectivity reduces the network gradient dispersion problem and enhances the overall network model stability. The effectiveness of this network model is demonstrated on the BCI Competition IV dataset 2a by integrating CNN, BiLSTM and ResNet (called CLRNet in this study) to decode motor imagery EEG. The network model combining CNN and BiLSTM achieved 87.0% accuracy in classifying motor imagery patterns in four classes. The network stability is enhanced by adding ResNet for cross-layer connectivity, which further improved the accuracy by 2.0% to achieve 89.0% classification accuracy. The experimental results show that CLRNet has good performance in decoding the motor imagery EEG dataset. This study provides a better solution for motor imagery EEG decoding in brain-computer interface technology research.

The survival rates and risk factors of implants in the early stage: a retrospective study.

BACKGROUND: Few large-sample studies in China have focused on the early survival of dental implants. The present study aimed to report the early survival rates of implants and determine the related influencing factors. METHODS: All patients receiving dental implants at our institution between 2006 and 2017 were included. The endpoint of the study was early survival rates of implants, according to gender, age, maxilla/mandible, dental position, bone augmentation, bone augmentation category, immediate implant, submerged implant category, implant diameter, implant length, implant torque, and other related factors. Initially, SPSS22.0 was used for statistical analysis. The Chi-square test was used to screen all factors, and those with p < 0.05 were further introduced into a multiple logistic regression model to illustrate the risk factors for early survival rates of implants. RESULTS: In this study, we included 1078 cases (601 males and 477 females) with 2053 implants. After implantation, 1974 implants were retained, and the early survival rate was 96.15%. Patients aged 30-60 years (OR 2.392), with Class I bone quality (OR 3.689), bone augmentation (OR 1.742), immediate implantation (OR 3.509), and implant length < 10 mm (OR 2.972), were said to possess risk factors conducive to early survival rates. CONCLUSIONS: The early survival rate of implants in our cohort exceeded 96%, with risk factors including age, tooth position, bone quality, implant length, bone augmentation surgery, and immediate implantation. When the above factors coexist, implant placement should be treated carefully.

Aloe-emodin induces apoptosis in human oral squamous cell carcinoma SCC15 cells.

BACKGROUND: Oral and pharyngeal cancer is the most common malignant human cancers. Chemotherapy is an effective approach for anti-oral cancer therapy, while the drug tolerance and resistance remain a problem for oral cancer patients. Aloe-emodin, rhein and physcion are classified as anthraquinones, which are the main pharmacodynamic ingredients of Rheum undulatum L.. This study was undertaken to investigate whether aloe-emodin, rhein and physcion show inhibiting growth and inducing apoptosis in oral squamous cell carcinoma SCC15 cells. We found that aloe-emodin show inhibiting growth and inducing apoptosis in oral squamous cell carcinoma SCC15 cells, we also investigated the underlying mechanisms of apoptosis induced by aloe-emodin. METHODS: Thiazolyl blue tetrazolium bromide (MTT) test was used to detect cell proliferation. Cell apoptosis was detected by flow cytometry. We also used western blot analysis to detect the potential mechanisms of apoptosis. RESULTS: Aloe-emodin, rhein and physcion inhibit the proliferation of SCC15 cells and the order of inhibition level are aloe-emodin > Rhein > Physcion, the half maximal inhibitory concentrations (IC50) value of aloe-emodin was 60.90 µM at 48 h of treatment. Aloe-emodin treatment resulted in a time- and dose-dependent decrease in cell viability and increased the apoptotic cell ratio. The results of western blotting showed the expression levels of caspase-9 and caspase-3 proteins increased following aloe-emodin treatment. CONCLUSIONS: Our results revealed that aloe-emodin treatment could inhibit cell viability of SCC15 cells and the potential mechanism of inhibition might be through the induction of apoptosis by regulation of the expression levels of caspase-9 and caspase-3. This indicates that aloe-emodin may be a good agent for anti-oral cancer drug exploring.

Pathogenic factors of maxillary sinus mucosal thickening observed by cone-beam computed tomography (CBCT).

OBJECTIVE: To explore the pathogenic factors associated with maxillary sinus mucosal thickening with Cone-beam computed Tomography (CBCT). METHODS: From 2016 through 2020, 93 patients with periapical periodontitis or periodontitis in the maxillary posterior dental region were selected. RESULTS: The preoperative thickness of the periodontitis group was significantly higher than that of the periapical periodontitis group (P < 0.05). The difference achieves statistical significance for the comparison of the thickness change with various severity of inflammation (F = 54.824, P = 0.000), the change with time (F = 312.741, P = 0.000). and the change with the interaction severity of inflammation and time（F = 86.132, P = 0.000). CONCLUSIONS: Patients with maxillary sinus mucosa thickening caused by periodontitis and periapical periodontitis should be extracted their infectious teeth and get thoroughly debridement. Maxillary sinus augmentation can perform favorable efforts 3-6 months after extracting teeth.

Treatment of Sinus Graft Infection After Sinus Floor Elevation: A Series of Four Case Reports.

This report of cases aims to share our treatment experiences in 4 sinus graft infection cases after sinus floor elevation and simultaneous implant placement. The preoperative and postoperative intraoral and radiographic photographs were collected and used to assess the treatment outcomes. The sinus cavity status, bone augmentation results, and implant stability were used as measurements to determine the treatment effectiveness. Four patients received partial graft removal as their surgical treatment for sinus graft infection combined with antibiotic therapy, with or without immediate secondary grafting. After early intervention, antibiotic therapy, and partial debridement of the infected sinus grafts, radiographic and clinical outcomes indicate successful resolution of the graft infection and stable bone graft levels around the implants. The keys to the successful management of the sinus graft infection were: early detection of the infection; early intervention, including partial debridement of the infected graft particles; and antibiotic therapy.

The clinical significance and application of the peri-implant phenotype in dental implant surgery: a narrative review.

Background and Objective: In recent years, the concept of the peri-implant phenotype has become a new standard for the clinical evaluation of the soft and hard tissues surrounding dental implants. Improving this phenotype enhances the likelihood of achieving long-term favorable results and is a necessary consideration during implant planning. Stable peri-implant tissue support is also crucial for the functional and aesthetic value of implant restoration. Herein, the authors review the clinical significance of the peri-implant phenotype and assess the timing of treatment strategies for improving peri-implant phenotype elements. Methods: A literature search was performed to retrieve papers on peri-implant tissue management and clinical outcomes published up to November 24th, 2022 in PubMed, Web of Science, EMBASE, and Scopus. Key Content and Findings: The optimal time to improve peri-implant bone thickness (PBT) is with augmentation procedures before implant surgery or at the same time as first-stage surgery. Similarly, issues associated with keratinized mucosa width (KMW) and mucosal thickness (MT) should be addressed before final restoration. The establishment of supracrestal tissue height (STH) depends on the MT and implant depth of the patient. Furthermore, special attention should be paid to the effect of the peri-implant phenotype on the prognosis of immediate implant placement in the aesthetic zone. Conclusions: The long-term success of implant restoration depends on careful planning that considers appropriate interventions for improving the peri-implant phenotype at different stages of treatment to reduce iatrogenic variables.

Temperature Changes in Oral All-Ceramic Materials with Different Optical Properties under Er:YAG Laser Irradiation.

Objectives: This in vitro study is aimed at assessing the oral all-ceramic materials energy transmission and temperature changes after Er:YAG laser irradiation of monolithic zirconia all-ceramic materials with varying optical properties. Materials and Methods: Two monolithic zirconia materials, Zenostar T and X-CERA TT (monolithic Zirconia), were studied. Specimens were divided into four groups, with a thickness of 1.0, 1.5, 2.0, and 2.5 mm, respectively. The chemical elemental composition of the two materials was determined using X-ray spectroscopy and Fourier transform infrared spectroscopy. The light transmittance of specimens with different thicknesses was measured using a spectrophotometer at three wavelength ranges: 200-380, 380-780, and 780-2500 nm. Irradiation with Er:YAG laser was performed, and the resultant temperature changes were measured using a thermocouple thermometer. Results: Compositional analysis indicated that Si content in X-CERA TT was higher than that in Zenostar T. The light transmittance of both materials decreased as specimen thickness increased. Er:YAG laser irradiation led to temperature increase at both Zenostar T (26.4°C-81.7°C) and X-CERA TT (23.9°C-53.5°C) specimens. Both optical transmittance and temperature changes after Er:YAG laser irradiation were consistent with exponential distribution against different thickness levels. Conclusion: Er:YAG laser penetration energy and resultant temperature changes were mainly determined by the thickness and composition of the examined monolithic zirconia materials.

Single cell atlas of developing mouse dental germs reveals populations of CD24+ and Plac8+ odontogenic cells.

The spatiotemporal relationships in high-resolution during odontogenesis remain poorly understood. We report a cell lineage and atlas of developing mouse teeth. We performed a large-scale (92,688 cells) single cell RNA sequencing, tracing the cell trajectories during odontogenesis from embryonic days 10.5 to 16.5. Combined with an assay for transposase-accessible chromatin with high-throughput sequencing, our results suggest that mesenchymal cells show the specific transcriptome profiles to distinguish the tooth types. Subsequently, we identified key gene regulatory networks in teeth and bone formation and uncovered spatiotemporal patterns of odontogenic mesenchymal cells. CD24+ and Plac8+ cells from the mesenchyme at the bell stage were distributed in the upper half and preodontoblast layer of the dental papilla, respectively, which could individually induce nonodontogenic epithelia to form tooth-like structures. Specifically, the Plac8+ tissue we discovered is the smallest piece with the most homogenous cells that could induce tooth regeneration to date. Our work reveals previously unknown heterogeneity and spatiotemporal patterns of tooth germs that may lead to tooth regeneration for regenerative dentistry.

Clinical Outcomes of Alveolar Ridge Augmentation with In Situ Autogenous Block Bone: A Retrospective Review.

PURPOSE: To present clinical outcomes of alveolar ridge augmentation using in situ autogenous block bone and to compare the outcomes with previous studies. MATERIALS AND METHODS: The medical records of patients with a severe horizontal bone defect in a partially edentulous alveolar ridge (width < 3.5 mm), who received bone augmentation using in situ autogenous block bone, were retrospectively reviewed. After a 6-month or longer healing period, the augmentation effect was examined before implant placement. Cone beam computed tomography (CBCT) was performed before and after surgeries. The alveolar width of the bone grafts was measured on the CBCT images. RESULTS: A total of 16 patients (22 grafts) were included. Graft exposure was seen in three grafts, which were classified as failed cases. The augmentation volume at implant placement in the failed cases was significantly lower than that of the successful cases. There were no significant differences in augmentation between anterior maxillary and mandibular implant sites. CONCLUSION: Autogenous bone grafting using in situ block bone is an effective and reliable approach for horizontal bone augmentation in the mandible and anterior maxilla that eliminates second donor site morbidity. Complete release of the buccal flap and tension-free suture is the key to avoiding wound dehiscence and ensuring the effectiveness of bone augmentation.

Conditioned medium derived from 3D tooth germs: A novel cocktail for stem cell priming and early in vivo pulp regeneration.

OBJECTIVES: Conditioned medium (CM) from 2D cell culture can mitigate the weakened regenerative capacity of the implanted stem cells. However, the capacity of 3D CM to prime dental pulp stem cells (DPSCs) for pulp regeneration and its protein profile are still elusive. We aim to investigate the protein profile of CM derived from 3D tooth germs, and to unveil its potential for DPSCs-based pulp regeneration. MATERIALS AND METHODS: We prepared CM of 3D ex vivo cultured tooth germ organs (3D TGO-CM) and CM of 2D cultured tooth germ cells (2D TGC-CM) and applied them to prime DPSCs. Influences on cell behaviours and protein profiles of CMs were compared. In vivo pulp regeneration of CMs-primed DPSCs was explored using a tooth root fragment model on nude mice. RESULTS: TGO-CM enhanced DPSCs proliferation, migration, in vitro mineralization, odontogenic differentiation, and angiogenesis performances. The TGO-CM group generated superior pulp structures, more odontogenic cells attachment, and enhanced vasculature at 4 weeks post-surgery, compared with the TGC-CM group. Secretome analysis revealed that TGO-CM contained more odontogenic and angiogenic growth factors and fewer pro-inflammatory cytokines. Mechanisms leading to the differential CM profiles may be attributed to the cytokine-cytokine receptor interaction and PI3K-Akt signalling pathway. CONCLUSIONS: The unique secretome profile of 3D TGO-CM made it a successful priming cocktail to enhance DPSCs-based early pulp regeneration.

Periodontitis regulates renal impairment in obese mice via TGF-ß/Smad pathway.

OBJECTIVE: To determine the impact of periodontitis on renal impairment induced by obesity. METHODS: Periodontitis and obesity models were induced using silk ligatures with bacteria and high-fat diet, respectively. Indicators of renal function were compared. Renal tubular epithelial cells (RTECs) were treated with lipopolysaccharides from periodontal pathogens in a high-fat environment to induce cell models of periodontitis and obesity. The transforming growth factor-ß/mothers against decapentaplegic homolog (Smad) (TGF-ß/Smad) pathway was evaluated both in vivo and in vitro. The indicators of renal function, renal pathological changes, and serum inflammatory cytokines were measured. The viability/apoptosis of RTECs and the expression of inflammatory cytokines were determined. RESULTS: Periodontitis resulted in an increase in TGF-ß/Smad activity in the kidney of obese mice. Moreover, the activity of RTECs was also increased in vitro. Downregulation of TGF-ß led to reduced TGF-ß, p-Smad2, p-Smad3, and Smad7 levels in kidney tissue and RTECs, ameliorated renal function indicators and renal pathological changes, increased viability and apoptosis of RTECs, and decreased levels of inflammatory cytokines. CONCLUSION: Periodontitis regulates renal impairment via the TGF-ß/Smad pathway in obese mice.

Impact of High-Altitude Hypoxia on Early Osseointegration With Bioactive Titanium.

Nowadays, the bone osseointegration in different environments is comparable, but the mechanism is unclear. This study aimed to investigate the osseointegration of different bioactive titanium surfaces under normoxic or high-altitude hypoxic environments. Titanium implants were subjected to one of two surface treatments: (1) sanding, blasting, and acid etching to obtain a rough surface, or (2) extensive polishing to obtain a smooth surface. Changes in the morphology, proliferation, and protein expression of osteoblasts on the rough and smooth surfaces were examined, and bone formation was studied through western blotting and animal-based experiments. Our findings found that a hypoxic environment and rough titanium implant surface promoted the osteogenic differentiation of osteoblasts and activated the JAK1/STAT1/HIF-1α pathway in vitro. The animal study revealed that following implant insertion in tibia of rabbit, bone repair at high altitudes was slower than that at low altitudes (i.e., in plains) after 2weeks; however, bone formation did not differ significantly after 4weeks. The results of our study showed that: (1) The altitude hypoxia environment would affect the early osseointegration of titanium implants while titanium implants with rough surfaces can mitigate the effects of this hypoxic environment on osseointegration, (2) the mechanism may be related to the activation of JAK1/STAT1/HIF-1α pathway, and (3) our results suggest the osteogenesis of titanium implants, such as oral implants, is closely related to the oxygen environment. Clinical doctors, especially dentists, should pay attention to the influence of hypoxia on early osseointegration in patients with high altitude. For example, it is better to choose an implant system with rough implant surface in the oral cavity of patients with tooth loss at high altitude.

LncRNA ELF3-AS1 is involved in the regulation of oral squamous cell carcinoma cell proliferation by reprogramming glucose metabolism.

PURPOSE: The present study aims to investigate the role of ELF3-AS1 in oral squamous cell carcinoma (OSCC). PATIENTS AND METHODS: A total of 112 patients with OSCC were admitted in Guangdong Provincial Stomatological Hospital from March 2016 to March 2019. RT-qPCR, cells and transient transfections, cell proliferation rate measurements and Western blots were carried out to analyze the samples. RESULTS: In the present study, we showed that ELF3-AS1 and glucose transporter 1 (GLUT1) were both upregulated in OSCC tissues, and those two factors were positively correlated. In OSCC cells, ELF3-AS1 overexpression resulted in upregulation, while ELF3-AS1 siRNA silencing caused downregulated expression of GLUT1 and glucose uptake. ELF3-AS1 and GLUT1 overexpression resulted in increased rate of OSCC cells, while ELF3-AS1 and GLUT1 siRNA silencing resulted in decreased proliferation rate of OSCC cells. In addition, GLUT1 siRNA silencing attenuated the effects of ELF3-AS1 overexpression. CONCLUSION: Therefore, ELF3-AS1 promotes the proliferation of OSCC cells by reprogramming glucose metabolism.

MALAT1 overexpression promotes the proliferation of human periodontal ligament stem cells by upregulating fibroblast growth factor 2.

Fibroblast growth factor 2 (FGF2) has been revealed to promote human periodontal ligament stem cell (PDLSC) proliferation. The abnormal proliferation of PDLSCs has also been associated with the pathogenesis of periodontitis. The long non-coding RNA, metastasis-associated lung adenocarcinoma transcript 1 (MALAT1), has been demonstrated to regulate FGF2 secretion. Therefore, MALAT1 may also be associated with periodontitis. The aim of the present study was to investigate the effect of MALAT1 overexpression on the proliferation of PDLSCs. In the current study, PDLSCs derived from healthy and periodontitis-affected teeth were collected. MALAT1 and FGF2 mRNA expression in PDLSCs was detected using reverse transcription-quantitative PCR. PDLSCs overexpressing MALAT1 were subsequently generated. PDLSC proliferation was analyzed using a Cell Counting kit-8 assay. FGF2 protein expression was detected using western blot analysis. The results revealed that MALAT1 and FGF2 mRNA were significantly upregulated in PDLSCs derived from periodontitis-affected teeth when compared with PDLSCs derived from healthy teeth. PDLSCs derived from periodontitis-affected teeth also demonstrated a significantly higher proliferation rate than PDLSCs derived from healthy teeth. MALAT1 and FGF2 mRNA expression were positively correlated in both PDLSC groups. MALAT1 overexpression promoted the proliferation of healthy and periodontitis-affected PDLSC groups and upregulated FGF2 protein expression. The present study concluded that MALAT1 overexpression promoted the proliferation of human PDLSC potentially via upregulating FGF2.

Wnt7a predicts poor prognosis, and contributes to growth and metastasis in tongue squamous cell carcinoma.

Regional and distant metastases are the principal reasons underlying the high mortality rate associated with tongue squamous cell carcinoma (TSCC); however, the precise molecular mechanisms involved in tongue tumorigenesis remain unknown. The present study aimed to determine the expression and mechanism of regulation of Wnt7a in the growth and metastasis of TSCC. Wnt7a mRNA and protein expression levels were examined in TSCC tissues using reverse transcription­quantitative polymerase chain reaction and immunohistochemical staining. A loss­of­function assay was performed in TSCC cell lines using Wnt7a small interfering RNA or short hairpin RNA, after which, cell proliferation, migration and invasion were analyzed using Cell Counting Kit­8, tumorigenicity and Transwell assays, respectively. Epithelial­mesenchymal transition (EMT)­associated proteins were detected by western blotting. The mRNA and protein expression levels of Wnt7a were significantly upregulated in cancer tissues compared with in the adjacent non­cancerous tissues. Clinical analysis indicated that Wnt7a expression was associated with T classification, lymph node metastasis and pathological differentiation, and high Wnt7a expression predicted a short recurrence­free survival for patients with TSCC. Silencing Wnt7a expression suppressed cell proliferation, migration and invasion, and reversed the EMT phenotype in TSCC cell lines. The present study revealed that Wnt7a may be upregulated in TSCC, where it may participate in modulating cell proliferation, migration, invasion and the EMT of TSCC. Therefore, Wnt7a should be considered a novel oncogene, and a potential prognostic and therapeutic target for patients with TSCC.

Potential functions of long non­coding RNAs in the osteogenic differentiation of human bone marrow mesenchymal stem cells.

Long non­coding RNAs (lncRNAs) are a specific group of RNA molecules that do not encode proteins. They have been shown to serve important regulatory functions in various biological and cell differentiation processes. However, the potential functions and regulatory mechanisms of lncRNAs that are associated with the osteogenic differentiation of human bone marrow mesenchymal stem cells (hBMSCs) remain to be elucidated. The present study aimed to investigate lncRNAs that are differentially expressed during the osteogenic differentiation of hBMSCs, along with the potential functions of those lncRNAs. To this end, three groups of hBMSCs were stimulated to undergo osteogenic differentiation for 7 days. Known lncRNAs, unknown lncRNAs and mRNAs that demonstrated differential expression prior to and following the osteogenic differentiation of hBMSCs were screened using lncRNA high­throughput sequencing. In addition, 12 lncRNAs were selected for reverse transcription­quantitative polymerase chain reaction (RT­qPCR) validation of the accuracy of the sequencing results. The potential functions and possible targets of the differentially expressed lncRNAs were analyzed using bioinformatics technologies (gene ontology, Kyoto Encyclopedia of Genes and Genomes and gene co­expression network analysis). In total, 64 lncRNAs were differentially expressed by at least two­fold in hBMSCs prior to and following osteogenic differentiation; these included seven known lncRNAs (two upregulated and five downregulated lncRNAs) and 57 unknown lncRNAs (35 upregulated and 22 downregulated lncRNAs). In addition, 409 mRNAs (257 upregulated and 152 downregulated mRNAs) were differentially expressed by at least two­fold. The RT­qPCR results obtained for 12 selected differentially expressed lncRNAs were consistent with the sequencing results. The gene co­expression network analysis of lncRNAs and mRNAs demonstrated that four lncRNAs (ENSG00000238042, lnc_1269, lnc_1369 and lnc_1708) may serve important roles in the osteogenic differentiation of hBMSCs. In conclusion, during the osteogenic differentiation of hBMSCs, the lncRNA expression profile changed significantly; certain of the observed differentially expressed lncRNAs may be derived from protein­coding genes and may serve important roles in osteogenic differentiation.

Silencing FOXC1 inhibits growth and migration of human oral squamous cell carcinoma cells.

Forkhead box C1 (FOXC1) is a transcription factor that serves an important role in regulating tumorigenesis and cancer progression. However, the expression and functional role of FOXC1 in oral squamous cell carcinoma (OSCC) remains unclear. FOXC1 protein expression was determined using immunohistochemical staining of OSCC tissues and normal tissues. Cell Counting Kit-8, colony formation, migration and 5-ethynyl-2'-deoxyuridine assays were performed to investigate the role and underlying mechanism of action of FOXC1 in OSCC. A consistent increase in the immunoreactive intensity of FOXC1 in OSCC tissues as compared with that in adjacent normal tissues was demonstrated. Knockdown of FOXC1 impaired cell growth and colony formation by inhibiting cell proliferation and reducing cyclin B1 and cyclin D1 levels in OSCC cells. FOXC1-silenced OSCC cells exhibited decreased migration compared with that demonstrated by the control cells, accompanied by a downregulation of matrix metalloproteinase (MMP)-2 and MMP-9. Collectively, the results of the present study demonstrated that FOXC1 functions as an oncogene in OSCC and may be an important therapeutic target and predictive biomarker for OSCC.

Analysis of the miRNA and mRNA involved in osteogenesis of adipose-derived mesenchymal stem cells.

Mesenchymal stem cells (MSCs) are bone marrow stromal cells capable of differentiating into different tissue types. Osteoblastic differentiation is a complex process that is critical for bone formation. An increasing number of studies have suggested that microRNAs (miRNAs) may serve important roles in various biological processes, including osteogenesis of MSCs. However, less is known about the participation of particular miRNAs in the osteogenic differentiation of adipose-derived stem cells (ADSCs). In order to identify functional miRNAs and the key genes involved in the osteogenesis of MSCs, the present study reconstructed a global network using data from the National Center for Biotechnology Information Gene Expression Omnibus. Meanwhile, gene ontology and pathway analysis were performed using the Cytoscape plug-in BinGO and the Database for Annotation, Visualization, and Integration Discovery, respectively. An miRNA-mRNA network composed of 72 mRNA and nine miRNA nodes advised by bioinformatics analysis was constructed. These mRNAs and miRNAs were predicted to be involved in the regulation of osteogenic differentiation of ADSCs according to the gene microarray. In the present study, six miRNAs (miR-143-3p, miR-135a-5p, miR-31-5p, miR-22-3p, miR-193b-3p and let-7i-5p) were observed to be highly associated with the osteogenesis of ADSCs, and dihydropyrimidinase like 3 was identified as a novel regulator in this process. These results provide support for further investigations into the management of bone regeneration-associated diseases.

[Effect of macrophage inflammatory protein-1ß on proliferation and apoptosis of human tongue squamous cell carcinoma CAL-27 cells in vitro].

OBJECTIVE: To detect CCR5 protein expression in different human tongue squamous cell carcinoma cells and observe the effect of macrophage inflammatory protein-1ß (MIP-1ß) on the proliferation and apoptosis of CAL-27 cells. METHODS: Western blotting and immunofluorescence staining were used to detect the expression of the CCR5, the receptor of MIP-1ß, in 3 human tongue squamous cell carcinoma cells UM-1, CAL-27, and Tca-8113. CCK-8 assay was used to assess the proliferation of CAL-27 cells stimulated with 10, 20, and 40 ng/mL MIP-1ß for 12, 24, or 48 h. The apoptosis of the cells stimulated with MIP-1ß (10, 20, and 40 ng/mL) for 24 h was analyzed using flow cytometry with Annexin V/PI double staining. RESULTS: CCR5 expression was detected both on the membrane and in the cytoplasm in all the 3 tongue squamous cell carcinoma cell lines. At the concentrations of 10, 20, and 40 ng/mL, MIP-1ß stimulation for 12 and 24 h significantly promoted the proliferation of CAL-27 cells (P<0.05); MIP-1ß stimulation for 48 h at the concentrations 10 and 20 ng/mL, but not at 40 ng/mL, promoted the proliferation of CAL-27 cells (P<0.05). MIP-1ß stimulation at 40 ng/mL for 24 produced the most obvious apoptosis-inducing effect in CAL -27 cells (P<0.05), while MIP-1ß at 10 or 20 ng/mL did not induce obvious apoptosis in the cells (P>0.05). CONCLUSION: CCR5 is expressed in all the 3 human tongue squamous cell carcinoma cells. MIP-1ß can promote the proliferation of CAL-27 cells but high concentrations of MIP-1ß also induced cell apoptosis. Prolonged stimulation of the cells with a high concentration of MIP-1ß shows attenuated effect in promoting cell proliferation probably as a result of cell apoptosis induced by MIP-1ß.

Investigation of proliferation and migration of tongue squamous cell carcinoma promoted by three chemokines, MIP-3α, MIP-1ß, and IP-10.

The aim of this work was to investigate the role of chemokines in proliferation and migration of tongue squamous cell carcinoma (TSCC). Out of the 80 cytokines surveyed by a human cytokine antibody array, three chemokines, macrophage inflammatory protein-3α (MIP-3α), macrophage inflammatory protein-1ß (MIP-1ß), and interferon gamma-induced protein 10 (IP-10), showed elevated expression in TSCC cells (CAL-27 and UM-1), compared to the oral mucosal epithelial cells. Immunohistochemistry confirmed the high level of expression of MIP-3α in the TSCC tissues, especially in the high clinical stages. Furthermore, Western blot and immunofluorescence staining indicated that C-C chemokine receptor type 5, C-C chemokine receptor type 6, and C-X-C motif chemokine receptor 3, which are the receptors for MIP-3α, MIP-1ß, and IP-10, respectively, were expressed in the TSCC cells. Viability assay showed MIP-3α, MIP-1ß, and IP-10 led to the proliferation of the CAL-27 cells. Interestingly, MIP-1ß and IP-10 also induced apoptosis in the TSCC cells. Transwell invasion assay showed MIP-3α and IP-10 could increase the invasive capability of TSCC cells; consistently, the enzymatic activities of matrix metalloproteinase-2 and matrix metalloproteinase-9 increased in the MIP-3α- and IP-10-treated cells. In summary, our results indicate the expression of MIP-3α, MIP-1ß, and IP-10 increased in the TSCC cells. The elevated expression of MIP-3α and IP-10 promoted proliferation and migration of TSCC. These chemokines, along with their receptors, could be potential biomarkers and therapeutic targets for TSCC, especially for those in the high clinical stages.