Ribosomal Dysregulation in Metastatic Laryngeal Squamous Cell Carcinoma: Proteomic Insights and CX-5461's Therapeutic Promise.

One of the main barriers to the successful treatment of laryngeal squamous cell carcinoma (LSCC) is postoperative progression, primarily due to tumor cell metastasis. To systematically investigate the molecular characteristics and potential mechanisms underlying the metastasis in laryngeal cancer, we carried out a TMT-based proteomic analysis of both cancerous and adjacent non-cancerous tissues from 10 LSCC patients with lymph node metastasis (LNM) and 10 without. A total of 5545 proteins were quantified across all samples. We identified 57 proteins that were downregulated in LSCC with LNM, which were enriched in cell adhesion pathways, and 69 upregulated proteins predominantly enriched in protein production pathways. Importantly, our data revealed a strong correlation between increased ribosomal activity and the presence of LNM, as 18 ribosomal subunit proteins were found to be upregulated, with RPS10 and RPL24 being the most significantly overexpressed. The potential of ribosomal proteins, including RPS10 and RPL24, as biomarkers for LSCC with LNM was confirmed in external validation samples (six with LNM and six without LNM) using Western blotting and immunohistochemistry. Furthermore, we have confirmed that the RNA polymerase I inhibitor CX-5461, which impedes ribosome biogenesis in LSCC, also decreases the expression of RPS10, RPL24, and RPS26. In vitro experiments have revealed that CX-5461 moderately reduces cell viability, while it significantly inhibits the invasion and migration of LSCC cells. It can enhance the expression of the epithelial marker CDH1 and suppress the expression of the mesenchymal markers CDH2, VIM, and FN at a dose that does not affect cell viability. Our study broadens the scope of the proteomic data on laryngeal cancer and suggests that ribosome targeting could be a supplementary therapeutic strategy for metastatic LSCC.

Immediate implant placement in the posterior mandibular region was assisted by dynamic real-time navigation: a retrospective study.

BACKGROUND: Efficient utilization of residual bone volume and the prevention of inferior alveolar nerve injury are critical considerations in immediate implant placement (IIP) within the posterior mandibular region. Addressing these challenges, this study focuses on the clinical efficacy and implant accuracy of dynamic real-time navigation, an emerging technology designed to enhance precision in implantation procedures. METHODS: This study included 84 patients with 130 implants undergoing immediate placement in the posterior mandibular region. Stratified into dynamic navigation, static guide plate, and freehand implant groups, clinical indicators, including initial stability, distance to the inferior alveolar nerve canal, depth of implant placement, and various deviations, were systematically recorded. Statistical analysis, employing 1- or 2-way ANOVA and Student's t-test, allowed for a comprehensive evaluation of the efficacy of each technique. RESULTS: All 130 implants were successfully placed with an average torque of 22.53 ± 5.93 N.cm. In the navigation group, the distance to the inferior alveolar nerve and the depth of implant placement were significantly greater compared to the guide plate and freehand groups (P < 0.05). Implant deviation was significantly smaller in both the navigation and guide plate groups compared to the freehand group(P < 0.05). Additionally, the navigation group exhibited significantly reduced root and angle deviations compared to the guide plate group(P < 0.05), highlighting the superior precision of navigation-assisted immediate implant placement. CONCLUSIONS: It is more advantageous to use dynamic navigation rather than a static guide plate and free-hand implant insertion for immediate posterior mandibular implant implantation.

Transcriptome analysis of osteogenic differentiation of human maxillary sinus mesenchymal stem cells using RNA-Seq.

Recent studies have demonstrated that human maxillary sinus mesenchymal stem cells (hMSMSCs) have osteogenic potential and can be osteogenically induced. Here, we investigated pivotal molecular functions and candidates that contribute to the osteogenic differentiation of hMSMSCs. Human maxillary sinus membranes were harvested from 3 patients with jaw deformities. hMSMSCs from human maxillary sinus membranes were osteogenically induced for 0 or 21 days. Subsequently, their functional profiles were analysed by RNA sequencing and validated by quantitative PCR. Compared with control hMSMSCs, osteogenically induced hMSMSCs showed (1) osteogenic differentiation phenotype, as evidenced by the cell nodes, alizarin red staining, osteogenesis-related protein, and RNA expression; (2) accelerated osteogenic process of ossification and calcium signalling, as demonstrated by Gene Ontology (GO) terms and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway; (3) enriched osteogenesis gene expression of SMOC2, OMD, IGF1, JUNB, BMP5, ADRA1A, and IGF2, which was validated by quantitative PCR. Based on by these results, we demonstrated that accelerated ossification process, calcium signalling, and upregulation of SMOC2, OMD, IGF1, JUNB, BMP5, ADRA1A and IGF2, may contribute to the osteogenic differentiation of hMSMSCs.

Anti-PD-L1 antibody enhances curative effect of cryoablation via antibody-dependent cell-mediated cytotoxicity mediating PD-L1highCD11b+ cells elimination in hepatocellular carcinoma.

Cryoablation (CRA) and microwave ablation (MWA) are two main local treatments for hepatocellular carcinoma (HCC). However, which one is more curative and suitable for combining with immunotherapy is still controversial. Herein, CRA induced higher tumoral PD-L1 expression and more T cells infiltration, but less PD-L1highCD11b+ myeloid cells infiltration than MWA in HCC. Furthermore, CRA had better curative effect than MWA for anti-PD-L1 combination therapy in mouse models. Mechanistically, anti-PD-L1 antibody facilitated infiltration of CD8+ T cells by enhancing the secretion of CXCL9 from cDC1 cells after CRA therapy. On the other hand, anti-PD-L1 antibody promoted the infiltration of NK cells to eliminate PD-L1highCD11b+ myeloid cells by antibody-dependent cell-mediated cytotoxicity (ADCC) effect after CRA therapy. Both aspects relieved the immunosuppressive microenvironment after CRA therapy. Notably, the wild-type PD-L1 Avelumab (Bavencio), compared to the mutant PD-L1 atezolizumab (Tecentriq), was better at inducing the ADCC effect to target PD-L1highCD11b+ myeloid cells. Collectively, our study uncovered the novel insights that CRA showed superior curative effect than MWA in combining with anti-PD-L1 antibody by strengthening CTL/NK cell immune responses, which provided a strong rationale for combining CRA and PD-L1 blockade in the clinical treatment for HCC.

TREM2+ macrophages suppress CD8+ T-cell infiltration after transarterial chemoembolisation in hepatocellular carcinoma.

BACKGROUND & AIMS: The immune landscape of hepatocellular carcinoma (HCC) following transarterial chemoembolisation (TACE) remains to be clarified. This study aimed to characterise the immune landscape following TACE and the underlying mechanism of HCC progression. METHODS: Tumour samples from five patients with treatment-naive HCC and five patients who received TACE therapy were collected and subjected to single-cell RNA sequencing. Another 22 paired samples were validated using immunofluorescence staining and flow cytometry. To clarify the underlying mechanisms, in vitro co-culture experiments and two types of TREM2-KO/WT mouse models, namely, an HCC cell orthotopic injection model and a spontaneous HCC model, were used. RESULTS: A reduced number of CD8+ T cells and an increased number of tumour-associated macrophages (TAMs) were observed in the post-TACE microenvironment. TACE therapy reduced the cluster CD8_C4, which was highly enriched with tumour-specific CD8+ T cells of pre-exhausted phenotype. TREM2 was found to be highly expressed in TAMs following TACE, which was associated with a poor prognosis. TREM2+ TAMs secreted less CXCL9 but more galectin-1 than did TREM2- TAMs. Galectin-1 promoted PD-L1 overexpression in vessel endothelial cells, impeding CD8+ T cell recruitment. TREM2 deficiency also increased CD8+ T cell infiltration, which inhibited tumour growth in both in vivo HCC models. More importantly, TREM2 deficiency enhanced the therapeutic effect of anti-PD-L1 blockade. CONCLUSIONS: This study shows that TREM2+ TAMs play an important role in suppressing CD8+ T cells. TREM2 deficiency increased the therapeutic effect of anti-PD-L1 blockade by enhancing antitumour activity of CD8+ T cells. These findings explain the reasons for recurrence and progression after TACE and provide a new target for HCC immunotherapy after TACE. IMPACT AND IMPLICATIONS: Studying the immune landscape in post-TACE HCC is important to uncover the mechanisms of HCC progression. By using scRNA sequencing and functional assays, we discovered that both the number and function of CD8+ T cells are compromised, whereas the number of TREM2+ TAMs is increased in post-TACE HCC, correlating with worse prognosis. Moreover, TREM2 deficiency dramatically increases CD8+ T cell infiltration and augments the therapeutic efficacy of anti-PD-L1 blockade. Mechanistically, TREM2+ TAMs display lower CXCL9 and increased Gal-1 secretion than do TREM2- TAMs, with Gal-1 mediating the overexpression of PD-L1 in vessel endothelial cells. These results suggest that TREM2 could be a novel immunotherapeutic target for patients treated with TACE in HCC. This provides an opportunity to break the plateau of limited therapeutic effect. This study has the value of understanding the tumour microenvironment of post-TACE HCC and thinking a new strategy of immunotherapy in the field of HCC. It is therefore of key impact for physicians, scientists and drug developers in the field of liver cancer and gastrointestinal oncology.

Regulation of endothelial cells on the osteogenic ability of bone marrow mesenchymal stem cells in peri-implantitis.

OBJECTIVES: The relationship between bone resorption and angiogenesis in peri-implantitis remains to be studied. We constructed a Beagle dog model of peri-implantitis, and extracted bone marrow mesenchymal stem cells (BMSCs) and endothelial cells (ECs) for culture. The osteogenic ability of BMSCs in the presence of ECs was investigated through an in vitro osteogenic induction model, and its mechanism was initially explored. SUBJECTS AND METHODS: The peri-implantitis model was verified by ligation, bone loss was observed by micro-CT, and cytokines were detected by ELISA. The isolated BMSCs and ECs were cultured to detect the expression of angiogenesis, osteogenesis-related proteins, and NF-κB signaling pathway-related proteins. RESULTS: 8 weeks after surgery, the peri-implant gums were swollen, and micro-CT showed bone resorption. Compared with the control group, IL-1ß, TNF-α, ANGII and VEGF were markedly increased in the peri-implantitis group. In vitro studies found that the osteogenic differentiation ability of BMSCs co-cultured with IECs was decreased, and the expression of NF-κB signaling pathway-related cytokines was increased. CONCLUSION: Endothelial cells inhibit the osteogenic differentiation of bone marrow mesenchymal stem cells through NF-κB signaling in the environment of peri-implantitis, which may become a new target for the treatment of peri-implantitis.

Clinical study of dynamic real-time navigation assisted immediate implant without flapping in the esthetic zone.

OBJECTIVE: The purpose of this study is to investigate the clinical effect of Dynamic real-time navigation to assist immediate implant without flapping in the esthetic zone. METHODS: Eight patients who underwent immediate implantation in the aesthetic area were included. A total of 11 implants were implanted using dynamic real-time navigation system combined with non-flap technology. Clinical indicators including implant deviation, initial stability, alveolar bone absorption, implant success rate, pink esthetic score (PES), Papilla index score (PIS), and the thickness of labial side bone plate of the implant were recorded. RESULTS: The deviation between the actual implant position and the preoperative design was (0.76±0.08) mm at the top, (1.11±0.18) mm at the root, (0.90±0.16) mm at the depth, and (1.48±0.91)°at the Angle. ISO values of all implants were greater than 59. PES was greater than 8. PIS index was 2 or 3. The average alveolar bone absorption was (0.34±0.09) mm and the thickness of bone plate on the lip of implant was greater than 1.6 mm. The success rate of implantation was 100%. CONCLUSION: The use of dynamic real-time navigation assisted non-flap implantation in the aesthetic area can effectively reduce implant deviation and improve the aesthetic effect.

A comparative study of the morphology and molecular biology between the Schneiderian membrane and palatine mucoperiosteum.

Schneiderian membrane is an indispensable structure for osteogenesis under the sinus floor space after maxillary sinus floor elevation.Therefore,this study aimed to compare the Schneiderian membrane and palatine mucoperiosteum in various aspects to explore whether the Schneiderian membrane has a periosteal layer and osteogenic ability. Schneiderian membrane and palatine mucoperiosteum specimens were collected and stained with HE, Masson, and Sirius red. Immunofluorescence staining was used to observe the expression and localization of mesenchymal stem cells (MSCs). Then MSCs from two tissues were isolated,cultured, and identified. The expression of osteogenic markers OCN, RUNX2, and BMP2 ware detected by Western blotting and quantitative PCR after osteogenic differentiation.The morphological observations revealed both the Schneiderian membrane and palatine mucoperiosteum were composed of three layers.Immunofluorescence staining showed that the inner bone surface layer of the Schneiderian membrane was rich in MSCs, which was similar to the cambium layer of the palatine mucoperiosteum.In addition, MSCs from two tissues showed similar morphological phenotype. After further osteogenic induction of the two groups, the expression of BMP2, RUNX2, and OCN were significantly increased. This study provide a novel insight into that Schneiderian membrane is a mucoperiosteal membrane rich of MSCs, containing a periosteal layer and osteogenic ability similar to mucoperiosteum.

Topology-enhanced molecular graph representation for anti-breast cancer drug selection.

BACKGROUND: Breast cancer is currently one of the cancers with a higher mortality rate in the world. The biological research on anti-breast cancer drugs focuses on the activity of estrogen receptors alpha (ER[Formula: see text]), the pharmacokinetic properties and the safety of the compounds, which, however, is an expensive and time-consuming process. Developments of deep learning bring potential to efficiently facilitate the candidate drug selection against breast cancer. METHODS: In this paper, we propose an Anti-Breast Cancer Drug selection method utilizing Gated Graph Neural Networks (ABCD-GGNN) to topologically enhance the molecular representation of candidate drugs. By constructing atom-level graphs through atomic descriptors for each distinct compound, ABCD-GGNN can topologically learn both the implicit structure and substructure characteristics of a candidate drug and then integrate the representation with explicit discrete molecular descriptors to generate a molecule-level representation. As a result, the representation of ABCD-GGNN can inductively predict the ER[Formula: see text], the pharmacokinetic properties and the safety of each candidate drug. Finally, we design a ranking operator whose inputs are the predicted properties so as to statistically select the appropriate drugs against breast cancer. RESULTS: Extensive experiments conducted on our collected anti-breast cancer candidate drug dataset demonstrate that our proposed method outperform all the other representative methods in the tasks of predicting ER[Formula: see text], and the pharmacokinetic properties and safety of the compounds. Extended result analysis demonstrates the efficiency and biological rationality of the operator we design to calculate the candidate drug ranking from the predicted properties. CONCLUSION: In this paper, we propose the ABCD-GGNN representation method to efficiently integrate the topological structure and substructure features of the molecules with the discrete molecular descriptors. With a ranking operator applied, the predicted properties efficiently facilitate the candidate drug selection against breast cancer.

Application of Computer-Aided Design and Individualized Templates for Bilateral Zygomaticomaxillary Complex Fractures.

PURPOSE: Reduction of the bilateral zygomaticomaxillary complex (ZMC) fracture with individualized templates based on computer- aided surgical simulation system. To evaluate the practicality and accuracy of this approach in the treatment of bilateral ZMC fracture. METHODS: Sixteen patients with bilateral ZMC fractures were collected to create a study model. The authors reconstruct the ZMC on one side via the three-dimensional (3D) model, and then mirrored to the opposite side. Multiple individualized templates were made based on the 3D model, and used as intraoperative guidance to reduce fractures. After surgery, the facial symmetry and the position of zygoma were observed. The mouth opening, pupil level, and sensation of infraorbital nerve were evaluated. Some mark points on zygoma were measured and the postoperative horizontal asymmetry rate (H) was calculated. Besides, orbital height and width were measured. RESULTS: For all patients, the position of bilateral ZMC was basically restored. The patients with restriction of mouth opening all recovered to normal. The H values were less than 3.0% at all mark points. There was almost no difference in bilateral orbital width and height. Meanwhile, there was no significant difference between the preoperative measurements of the ideal virtual 3D model and the postoperative measurements of patients. CONCLUSIONS: The study proves that application of computer-aided design and individualized templates can accurately guide the reduction operation of ZMC fracture, restore the ideal shape of ZMC, and obtain good facial symmetry.

Distinctive microbiota of delayed healing of oral mucositis after radiotherapy of nasopharyngeal carcinoma.

Background: Oral mucositis is the most common complication after radiotherapy of nasopharyngeal carcinoma (NPC). Previous studies had revealed that oral microbiota took great alteration soon after and during radiotherapy. Here, we aimed to investigate if the alteration of oral microbiota was related to delayed healing of oral mucositis after six month of radiotherapy. Methods: We recruited 64 NPC patients and collected samples after six month of radiotherapy. 32 patients were included into normal healing group (N), 22 patients were mild delayed healing group (M), while 10 patients were severe delayed healing group (S). 16S rRNA gene sequencing was used to assess and identify oral microbiota alteration. Results: The diversity of oral microbial communities was not significantly different. Composition of oral microbial was huge different among S group, for the Actinobacteria and Veillonella were significantly increased, which showed significant dysbiosis of the oral microbiome. Functional analysis of metabolic pathways of oral microbiota demonstrated that degradation of organic acids and amino acids were significantly increased in S group. Moreover, phenotype analysis found that relative abundance of aerobic and biofilm formation were higher in S group. We also found the Actinobacteria co-occurred with Veillonellaceae, but anti-occurred with other biofilm oral bacteria. These two biomarkers may be predictable for severe delayed healing of oral mucositis after radiotherapy. Conclusion: This study suggests a potential association between oral microbiome and delayed healing of oral mucositis. The Actinobacteria and Veillonellaceae may be biomarkers in predicting the risks for the severe delayed healing of oral mucositis after radiotherapy of NPC.

Experts' consensus on perioperative management of tooth extractions in patients receiving oral antithrombotic treatment.

Thromboembolic diseases, which comprise venous thromboembolic diseases and arterial thromboembolic diseases, have become the number one cause of death worldwide. To prevent or treat thrombosis, patients with thromboembolic diseases need to take antithrombotic drugs, which would increase the risk of bleeding during and after surgery. Tooth extraction is the most common operation in oral and maxillofacial surgery clinics. Although patients given oral antithrombotic drugs do not need to undergo drug withdrawal, the perioperative management of such patients remains confusing to most clinicians. Moreover, the potential risk factors for bleeding warrant further study. To improve the clinicians' knowledge of perioperative management for patients subjected to tooth extractions with oral antithrombotic drugs, experts have drafted this consensus focusing on preoperative bleeding risk assessment, intraoperative operating norms, and postoperative care to summarize the points needing attention.

The diverse roles of YAP in the regulation of human nasal epithelial remodeling.

Yes-associated protein (YAP) is essential in maintaining tissue size. Aberrant epithelial remodeling is a key pathological alteration in both inflammation and benign tumors in nasal mucosa. We sought to investigate the expression and localization patterns of YAP in remodeled nasal epithelium of basal cell hyperplasia, goblet cell metaplasia and squamous metaplasia. YAP expression patterns were evaluated in tissues obtained from patients with NP (n = 45) and IP (n = 27), and control subjects with septal deviation (n = 17) and tissue-derived primary cell cultures. Compared to the normal epithelium, expressions of YAP were significantly higher in basal cell hyperplasia (NP, 11.4-fold; IP, 19.6-fold), followed by squamous metaplasia (8.2-fold) and mild to moderate goblet cell metaplasia (2.9-fold); while their expression was lower in severe goblet cell metaplasia (3.3-fold). Our resultsshowed that: 1) ectopic nuclear YAP expression associated with p63+ basal cell hyperplasia and the high proliferative potential epithelial cells; 2) increase of cytoplasmic YAP correlated with mild to moderate goblet cell metaplasia; 3) increase of cytoplasmic YAP correlated with squamous cell metaplasia. The in vitro cell model also demonstrated almost concordant changes of YAP with the mucosa findings. Different YAP expression and localization patterns should play critical but differential roles in the nasal epithelial remodeling processes under mucosal inflammation and benign tumor formation.

Chitosan as additive affects the bacterial community, accelerates the removals of antibiotics and related resistance genes during chicken manure composting.

Manures, storages for antibiotic resistance genes (ARGs), pollute soil and water as well as endanger human health. Recently, we have been searching a better solution to remove antibiotics and ARGs during aerobic composting. Here, the dynamics of chitosan addition on the profiles of 71 ARGs, bacterial communities, chlortetracycline (CTC), ofloxacin (OFX) were investigated in chicken manure composting and compared with zeolite addition. Chitosan addition effectively reduces antibiotics contents (CTC under detection limit, OFX 90.96%), amounts (18) and abundance (56.7%, 11.1% higher than zeolite addition) of ARGs and mobile genetic elements (MGEs) after 42 days composting. Network analysis indicated that a total of 27 genera strains assigned into 4 phyla (Firmicutes, Proteobacteria, Actinobacteria and Bacteroidetes) were the potential hosts of ARGs. Redundancy analysis (RDA) demonstrated that bacterial community succession is the main contributor in the variation of ARGs. Overall, chitosan addition may effect bacterial composition by influencing physic-chemical properties and the concentration of antibiotics, Cu2+, Zn2+ to reduce the risk of ARG transmission. This study gives a new strategy about antibiotics and ARGs removal from composting on the basis of previous studies.

Inorganic matter modified water-based copolymer prepared by chitosan-starch-CMC-Na-PVAL as an environment-friendly coating material.

Inorganic matter modifications were used to improve the hydrophobic properties and slow-release effects of water-based copolymer films. Water-based copolymers were prepared by aqueous polymerization of polyvinyl alcohol, starch, chitosan, and sodium carboxymethyl cellulose, and then, zeolite powder, volcanic ash or biochar were added to prepare the inorganic matter modified water-based copolymer films. The results showed that the inorganic matter modified water-based copolymer films had enhanced thermal stability, reductions in O-H and water vapour permeability, and increased crystallinity and roughness. Compared with water-based copolymer films, the water absorption capacities of the zeolite powder modified water-based copolymer films, volcanic ash modified water-based copolymer films, and biochar modified water-based copolymer films were reduced by 42.8 %, 50.0 % and 39.0 %, and their ammonium permeability was reduced by 53.0 %, 12.1 % and 1.1 %, respectively. Inorganic matter modified water-based copolymer films have properties that make them suitable for use in preparing slow-release coating materials.

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Pot experiment with winter wheat was conducted to investigate the effects of blended nitrogen (N) fertilizer (slow-release fertilizer-N:urea-N=1:1) combined with N fertilizer inhibitor NAM on soil ammonium (NH4+-N), nitrate (NO3--N), microbial biomass nitrogen (MBN) and fixed-ammonium (FN) contents. We analyzed dynamic characteristics of soil mineral N, MBN, FN pools under different treatments. There were six treatments, including no N fertilizer (CK), conventional urea (U), blended N fertilizer (MU), MU plus 2.5‰ NAM (MUN1), MU plus 5‰ NAM (MUN2), and MU plus 7.5‰ NAM (MUN3). Our results showed that, compared to that of MU treatment, MUN2 and MUN3 delayed the appearance time of NH4+-N peak. Averaged across the whole wheat growing period, soil mineral N content for NAM treatments decreased by 5.3%-11.7%. From tillering to maturity stage, MBN mineralization and mineralization rates were 38.96 mg·kg-1 and 91.5%, which was higher than that of U treatment; MBN mineralization and mineralization rates for MUN1, MUN2 and MUN3 treatments were 58.73 mg·kg-1, 83.3%, 94.20 mg·kg-1, 94.6%, 104.46 mg·kg-1 and 96.3%, respectively. The FA mineralization release for NAM treatments were higher by 2.83-9.19 mg·kg-1 than that of MU treatment. The results of path analysis showed that NAM addition weakened the direct effect of soil NH4+-N pool on NO3--N pool but enhanced the indirect effects of FN pool on NO3--N pool through affecting NH4+-N pool. The wheat grain yields of the MUN1, MUN2 and MUN3 treatments were significantly higher by 31.6%, 21.5% and 22.9% than that of MU treatment. Nitrogen use efficiencies were increased by 8.1%, 13.5% and 3.1%, respectively. In summary, through double regulation for N release and transformation in soil, NAM delayed the appearance time of soil NH4+-N peak and retarded its transformation into NO3--N, and increased the roles of MBN and FN in supplying N, thereby increased crop yield and N-fertilizer use efficiency.

Hsa_circRNA_33287 promotes the osteogenic differentiation of maxillary sinus membrane stem cells via miR-214-3p/Runx3.

BACKGROUND: Circular RNAs (circRNAs) comprise a novel class of noncoding RNAs that play important roles in a variety of diseases. However, the mechanism by which circRNAs regulate the osteogenic differentiation of maxillary sinus membrane stem cells (MSMSCs) remains largely unclear. METHODS: Microarray analysis was used to explore the expression profiles of circRNAs during the osteogenic differentiation of normal and BMP2 induced-MSMSCs. CircRNA_33287 was identified by agarose electrophoresis, quantitative real-time PCR (qRT-PCR), and western blotting. The function of circRNA_33287 was assessed by loss- and gain-of-function techniques and Alizarin red staining. Potential miRNA binding sites for circRNA_33287, and the target genes of miR-214-3p, were predicted by using online bioinformatics analysis tools. The relationships among the regulatory roles played by circRNA_33287, miR-214-3p, and Runt-related transcription factor 3 (Runx3), during the osteogenic differentiation of MSMSCs were verified by use of the dual luciferase reporter assay, qRT-PCR, and western blotting techniques, respectively. In addition, the molecular sponge potential of circRNA_33287 for miRNA was assessed via in vivo ectopic bone formation and a histological analysis performed after hematoxylin and eosin staining. RESULTS: Expression of circRNA_33287 was confirmed to be up-regulated during the osteogenic differentiation of MSMSCS. Overexpression and silencing of circRNA_33287 increased and decreased the expression levels of key markers of osteogenesis, respectively, including Runx2, OSX, and ALP. Furthermore, circRNA_33287 acted as a molecular sponge for miR-214-3p, which regulated Runx3 expression by targeting its 3'UTR. Moreover, circRNA_33287 protected Runx3 from miR-214-3p-mediated suppression. In addition, circRNA_33287 was shown to increase ectopic bone formation in vivo and displayed the strongest ability to stimulate bone formation when co-transfected with a miR-214-3p inhibitor. CONCLUSION: The novel pathway circRNA_33287/miR-214-3p/Runx3 was found to play a role in regulating the osteoblastic differentiation of MSMSCs in the posterior maxilla.

Lnc-NTF3-5 promotes osteogenic differentiation of maxillary sinus membrane stem cells via sponging miR-93-3p.

BACKGROUND: The function and the mechanism of long non-coding RNAs (lncRNAs) on the osteogenic differentiation of maxillary sinus membrane stem cells (MSMSCs) remain largely unknown. MATERIALS AND METHODS: The expression of lnc-NTF3-5 and Runt-related transcription factor 2 (RUNX2), Osterix (OSX), and Alkaline Phosphatase (ALP) was examined by quantitative real-time PCR (qRT-PCR) in MSMSCs during the process osteogenic differentiation. Then the function of lnc-NTF3-5 was evaluated by loss- and gain-of-function techniques, as well as qRT-PCR, western blot, and Alizarin Red staining. In addition, the microRNAs (miRNAs) sponge potential of lnc-NTF3-5 was assessed through RNA immunoprecipitation, dual luciferase reporter assay, and in vivo ectopic bone formation. RESULTS: Lnc-NTF3-5, RUNX2, OSX, and ALP increased alone with the differentiation. Inhibition of lnc-NTF3-5 decreased the expression of RUNX2, OSX, and ALP both at mRNA and protein levels. Alizarin red staining showed similar trend. In contrast, overexpression of lnc-NTF3-5 presented totally opposite effects. Besides, overexpression of lnc-NTF3-5 could decrease the expression of microRNA-93-3p (miR-93-3p). Enhance miR-93-3p could also inhibit the expression level of lnc-NTF3-5. RNA immunoprecipitation demonstrated that lnc-NTF3-5 is directly bound to miR-93-3p and dual luciferase reporter assay proved that miR-93-3p targets 3' UTR of RUNX2 to regulate its expression. Ultimately, in vivo bone formation study showed that lnc-NTF3-5 and miR-93-3p inhibitor co-transfection group displayed the strongest bone formation. CONCLUSIONS: The novel pathway lnc-NTF3-5/miR-93-3p/RUNX2 could regulate osteogenic differentiation of MSMSCs and might serve as a therapeutic target for bone regeneration in the posterior maxilla.

Preparation and characterization of slow-release fertilizer encapsulated by biochar-based waterborne copolymers.

To enhance the effectiveness of polyvinyl alcohol (PVA) coated fertilizers, a novel slow-release fertilizer was developed using biochar and waterborne copolymer of PVA and polyvinylpyrrolidone (PVP) as coating materials. The effects of botanical origins and doses of biochar as well as copolymer concentrations on biochar-copolymer structures and properties were investigated. Our results indicated that the water absorbency of blend films differing in biochar origins showed different responses to these three factors. Generally, biochar decreased water absorbency of copolymer with an increased degradability, and contributed to improving the slow-release property of coated urea. In particular, rice biochar-based copolymer (S5 film) had less hydrophilic OH bonds and encapsulated urea granules more compactly and densely. The urea particles coated with rice biochar-based copolymer (S5 film) exhibited an excellent release behavior of 65.28% nutrient leaching on the 22th day. Therefore, this work has demonstrated the potential of biochar-based copolymer from different botanical biochar for improving the effectiveness of fertilizers.

Endoscope-assisted neurectomy and inferior alveolar nerve avulsion in treating trigeminal neuralgia.

OBJECTIVE: To explore endoscope-assisted surgical treatment for idiopathic trigeminal neuralgia (TN), with neurectomy and avulsion of the inferior alveolar nerve (IAN), when patients are not willing to undergo intracranial procedures. CLINICAL MATERIALS AND METHODS: Five TN patients with inferior alveolar neuralgia in the mandibular branch were admitted; endoscope-assisted inferior alveolar neurectomy and avulsion was performed for TN treatment. Clinical observation and postoperative follow-up were conducted. RESULTS: After the operation, all patients had complete relief of pain, with mild postoperative swelling, limited mouth opening, and fast recovery. Patients were followed up for 3-24 months. Facial numbness and paresthesia were felt in the IAN distribution area of the operated side, with no recurrence. CONCLUSION: Endoscope-assisted neurectomy and avulsion of the IAN is a safe surgical method to treat TN in the mandibular branch when craniotomy is contraindicated or when patients decline neurosurgical options. It is effective in pain relief, with limited invasiveness and speedy recovery.