Disruption of hedgehog signaling leads to hyoid bone dysplasia during embryogenesis.

The development of the hyoid bone is a complex process that involves the coordination of multiple signaling pathways. Previous studies have demonstrated that disruption of the hedgehog pathway in mice results in a series of structural malformations. However, the specific role and critical period of the hedgehog pathway in the early development of the hyoid bone have not been thoroughly characterized. In this study, we treated pregnant ICR mice with the hedgehog pathway inhibitor vismodegib by oral gavage in order to establish a model of hyoid bone dysplasia. Our results indicate that administration of vismodegib at embryonic days 11.5 (E11.5) and E12.5 resulted in the development of hyoid bone dysplasia. We were able to define the critical periods for the induction of hyoid bone deformity through the use of a meticulous temporal resolution. Our findings suggest that the hedgehog pathway plays a crucial role in the early development of the hyoid bone. Additionally, our research has established a novel and easily established mouse model of synostosis in the hyoid bone using a commercially available pathway-selective inhibitor.

LINC00664/miR-411-5p/KLF9 feedback loop contributes to the human oral squamous cell carcinoma progression.

OBJECTIVES: Oral squamous cell carcinoma (OSCC) is one of the most aggressive head and neck cancers with high incidence. Multiple studies have revealed that long non-coding RNAs (lncRNAs) play pivotal roles in tumorigenesis. However, the role of long intergenic non-protein coding RNA 664 (LINC00664) on the progression of OSCC was still unclear. SUBJECTS AND METHODS: In this study, the expression of LINC00664 in OSCC tissues and cell lines was detected by quantitative real-time polymerase chain reaction (qRT-PCR). The functional role of LINC0664 was estimated by cell counting kit-8 (CCK-8), transwell assays, Western blot in vitro, and xenograft tumor model in vivo. The regulatory mechanism was investigated by RNA-binding protein immunoprecipitation (RIP), chromatin immunoprecipitation (ChIP), and luciferase reporter assays. RESULTS: LINC00664 was found to be upregulated in OSCC tissues and cell lines and was associated with poor prognosis of OSCC patients. LINC00664 knockdown suppressed OSCC cell proliferation, migration, invasion, and epithelial-mesenchymal transition (EMT). Moreover, Kruppel like factor 9 (KLF9) enhanced LINC00664 expression at transcription level. Interestingly, LINC00664 upregulated KLF9 expression by sponging miR-411-5p. In addition, knockdown of LINC00664 restrained tumor growth of OSCC in vivo. CONCLUSION: Our study identified the oncogenic roles of LINC00664 in OSCC tumorigenesis and EMT via KLF9/LINC00664/miR-411-5p/KLF9 feedback loop, which provides new perspectives of the potential therapeutic target for OSCC.

A Comparative Evaluation of Iliac Crest Cortical-Cancellous Bone Blocks Graft With and Without Concentrated Growth Factors (CGFs) in Secondary Alveolar Bone Grafting: A Retrospective Study.

BACKGROUND: The purpose of this study was to investigate the clinical effect and bone resorption of iliac crest cortical-cancellous bone block grafts combined with concentrated growth factor (CGF) compared with iliac crest cortical-cancellous bone block grafts only in secondary alveolar bone grafting. MATERIALS AND METHODS: Eighty-six patients (43 in the CGF group and 43 in the non-CGF group) with unilateral alveolar clefts were examined. Patients (17 in the CGF group and 17 in the non-CGF group) were randomly chosen for radiologic evaluation. Quantitative evaluation of the bone resorption rate was made with cone-beam computed tomography and Mimics 19.0 software at 1 week and 12 months after surgery. RESULTS: The success rate of bone grafting was 95.3% and 79.1% in the CGF and non-CGF groups, respectively ( P =0.025). The mean bone resorption rate at 12 months postoperatively was 35.66±15.80% and 41.39±19.57% in the CGF and non-CGF groups, respectively ( P =0.355). The bone resorption patterns of the 2 groups were similar on the labial, alveolar process, and palatal sides, and there was no obvious bone resorption on the labial side in either group. Nasal side bone resorption in the CGF group was significantly less than that in the non-CGF group ( P =0.047). CONCLUSIONS: Cortical-cancellous bone block grafts reduce labial bone resorption, while CGF reduces nasal bone resorption and improves the success rate. The combination of bone block and CGF in secondary alveolar bone grafting is worthy of further clinical application.

Revitalizing mouse diphyodontic dentition formation by inhibiting the sonic hedgehog signaling pathway.

BACKGROUND: Tooth regeneration depends on the longevity of the dental epithelial lamina. However, the exact mechanism of dental lamina regression has not yet been clarified. To explore the role of the Sonic hedgehog (Shh) signaling pathway in regression process of the rudimentary successional dental lamina (RSDL) in mice, we orally administered a single dose of a Shh signaling pathway inhibitor to pregnant mice between embryonic day 13.0 (E13.0) and E17.0. RESULTS: We observed that the Shh signaling pathway inhibitor effectively inhibited the expression of Shh signaling pathway components and revitalized RSDL during E15.0-E17.0 by promoting cell proliferation. In addition, mRNA-seq, reverse transcription plus polymerase chain reaction (RT-qPCR), and immunohistochemical analyses indicated that diphyodontic dentition formation might be related to FGF signal up-regulation and the Sostdc1-Wnt negative feedback loop. CONCLUSIONS: Overall, our results indicated that the Shh signaling pathway may play an initial role in preventing further development of mouse RSDL in a time-dependent manner.

MMP1 3'UTR facilitates the proliferation and migration of human oral squamous cell carcinoma by sponging miR-188-5p to up-regulate SOX4 and CDK4.

Growing evidence indicates that the non-coding 3'-untranslated region (3'UTR) of genes acts as competing endogenous RNAs (ceRNAs) to exert their roles in a number of diseases, including cancer. In the present study, MMP1 messenger RNA was identified to be significantly up-regulated in oral squamous cell carcinoma (OSCC) tissues, and both MMP1 and its 3'UTR promoted tumor growth and cell motility. Further mechanism investigations indicated that MMP1 3'UTR was able to antagonize miR-188-5p; in addition, overexpression of MMP1 3'UTR up-regulated the expression level of SOX4 and CDK4, target genes of miR-188-5p, which have also been identified as oncogenic driver genes in OSCC. Therefore, a ceRNA regulatory network among MMP1, SOX4, and CDK4 mediated via competing for binding to miR-188-5p was proved. Taken together, the present study demonstrates for the first time that MMP1 mRNA participates in the development of OSCC via ceRNA regulatory mechanism and genes involved in the ceRNA network may provide a novel avenue for target therapy.

Tumor-associated macrophages promote prostate cancer progression via exosome-mediated miR-95 transfer.

Tumor-associated macrophages (TAMs) are vital constituents in mediating cell-to-cell communication within the tumor microenvironment. However, the molecular mechanisms underlying the interplay between TAMs and tumor cells that guide cell fate are largely undetermined. Extracellular vesicles, also known as exosomes, which are derived from TAMs, are the components exerting regulatory effects. Thus, understanding the underlying mechanism of "onco-vesicles" is of crucial importance for prostate cancer (PCa) therapy. In this study, we analyzed micro RNA sequences in exosomes released by THP-1 and M2 macrophages and found a significant increase in miR-95 levels in TAM-derived exosomes, demonstrating the direct uptake of miR-95 by recipient PCa cells. In vitro and in vivo loss-of-function assays suggested that miR-95 could function as a tumor promoter by directly binding to its downstream target gene, JunB, to promote PCa cell proliferation, invasion, and epithelial-mesenchymal transition. The clinical data analyses further revealed that higher miR-95 expression results in worse clinicopathological features. Collectively, our results demonstrated that TAM-mediated PCa progression is partially attributed to the aberrant expression of miR-95 in TAM-derived exosomes, and the miR-95/JunB axis provides the groundwork for research on TAMs to further develop more-personalized therapeutic approaches for patients with PCa.

[Correlation analysis of color change and Maillard reaction during processing of Gardeniae Fructus Praeparatus].

The chromaticity space parameters of the samples during the processing of Gardeniae Fructus Praeparatus(Jiaozhizi in Chinese herbal name, JZZ) were measured by the visual analyzer to analyze the color variation rule during the processing of JZZ, and the content changes of total reducing sugar, total amino acid and 5-hydroxymethylfurfural(5-HMF) related to Maillard reaction were measured. Pearson correlation analysis and linear regression analysis of the data were carried out by SPSS 24.0 software. The experimental results showed that the objective coloration of the samples in the processing of JZZ was basically consistent with the traditional subjective color judgment; the contents of total reducing sugar and total amino acids showed a decreasing trend during the processing of JZZ, and the content of 5-HMF showed an increasing trend, which was in line with Maillard reaction law. Pearson correlation analysis results showed that there was a significant correlation between the chromaticity space parameters L~*(lightness value), a~*(red green value), b~*(yellow blue value), E~*ab(total color value) and the contents of total reducing sugar, total amino acid and 5-HMF(P<0.01), among which the values of L~*, a~*, b~*, E~*ab were positively correlated with the contents of total reducing sugar and total amino acid, and negatively correlated with the contents of 5-HMF. The results of linear regression analysis also showed that these two were highly correlated. In this study, by establishing the correspondence relationship between the color change of JZZ processing and Maillard reactants, wecan not only provide a basis for the objective digital expression of subjective color of JZZ, but also provide a reference for explaining the processing mechanism of JZZ from a new perspective.

Reciprocal regulation of miR-1254 and c-Myc in oral squamous cell carcinoma suppresses EMT-mediated metastasis and tumor-initiating properties through MAPK signaling.

AIM: This study aimed to determine the effect of miR-1254 on oral squamous cell carcinoma (OSCC) metastasis and the specific mechanism involved. METHODS: The metastatic properties of OSCC cells were analyzed by transwell assays. The tumor-initiating properties of OSCC cells were analyzed by tumor sphere formation assays. The mRNA and protein expressions of targeted genes were determined by quantitative polymerase chain reaction assays and western blot analyses, respectively. Xenograft experiments were employed to evaluate the anti-metastatic effects of miR-1254 and miR-1254-mediated cancer stem cell (CSC) properties in vivo. The gene targets of miR-1254 were investigated by luciferase reporter assays. Chromatin immunoprecipitation assays were performed to observe the transcriptional regulation of miR-1254 biogenesis by transcription factor. RESULTS: miR-1254 attenuated OSCC metastasis and tumor-initiating properties in vitro and in vivo. Consistent with the experimental observations, miR-1254 was decreased in late-stage OSCCs and strongly correlated with risk of OSCC metastasis. Moreover, miR-1254 was mechanistically shown to down-regulate MAP3K3, accompanied by inactivation of the MAPK signaling pathway and inhibition of epithelial-mesenchymal transition (EMT) in OSCC cells. miR-1254 was transcriptionally repressed by c-Myc to form a positive feed back loop through MAPK signaling. CONCLUSION: Our findings suggest that miR-1254 is a potential target for the treatment of OSCCs, and miR-1254 can be clinically utilized as a biomarker for the clinical prognosis or diagnosis of OSCCs.

Disruption of Hedgehog Signaling by Vismodegib Leads to Cleft Palate and Delayed Osteogenesis in Experimental Design.

The function of hedgehog signaling has previously been shown to be crucial for craniofacial development. In this study, we treated C57/BL6J mice with the hedgehog pathway inhibitor vismodegib by oral gavage to establish a stable vismodegib-induced cleft palate model. At E10.5 and E12.5, mice in the experimental group were treated with 100 mg/kg of vismodegib, whereas mice in the control group were treated with solvent. The treated pregnant mice were sacrificed on E13.5, E14.5, E15.5, and E16.5. Palatal shelf growth was evaluated via histological and immunohistochemical analyses as well as palatal organ culture. Immunohistochemical staining was performed to examine the expression of osteogenic proteins in the palatal tissue. A high proportion of the mice administered 2 doses of 100 mg/kg of vismodegib displayed a cleft palate. Histologic examination revealed severely retarded palatal shelf growth and thickened epithelium in the experimental group. Vismodegib exposure induced complete cleft palate, which was attributed to a reduced cell proliferation rate in the palatal mesenchyme along the anterior-posterior axis. Moreover, this model also showed delayed ossification in the region of palatine bone with downregulation of Indian hedgehog (Ihh) protein. Our results suggest that vismodegib can be used to inhibit hedgehog signaling to affect palatal morphogenesis. Under treatment with this exogenous inhibitor, the cell proliferation rate of the palatal shelves and the osteogenic potential of the hard palate were decreased, which likely contributed to the complete cleft palate.

Iron-Catalyzed Intermolecular 1,2-Difunctionalization of Styrenes and Conjugated Alkenes with Silanes and Nucleophiles.

The first iron-catalyzed 1,2-difunctionalization of styrenes and conjugated alkenes with silanes and either N or C, using an oxidative radical strategy, is described. Employing FeCl2 and di-tert-butyl peroxide allows divergent alkene 1,2-difunctionalizations, including 1,2-aminosilylation, 1,2-arylsilylation, and 1,2-alkylsilylation, which rely on a wide range of nucleophiles, namely, amines, amides, indoles, pyrroles, and 1,3-dicarbonyls, thus providing a powerful platform for producing diverse silicon-containing alkanes.

TRIM47 silencing inhibits the malignant biological behaviors of prostate cancer cells by regulating MDM2/p53 signaling.

Prostate cancer (PCa) is a prevalent male malignancy globally. Tripartite motif 47 (TRIM47) has been reported to be associated with PCa. However, how TRIM47 acts on PCa is still incompletely understood. Here, we explored the biological roles of TRIM47 in PCa cells and investigated its potential regulatory mechanism. TRIM47 expression in PCa cells was detected by qRT-PCR and western blot. After TRIM47 silencing, the viability of PCa cells was measured using CCK-8 method. Flow cytometry was employed to estimate cell cycle. Cell apoptotic level was subjected to appraisement with TUNEL assay. Additionally, wound healing- and transwell assays were adopted for evaluation of migration and invasion of PCa cells. Moreover, the Biogrid database and HDOCK SERVER predicated that TRIM47 could interact with mouse double minute 2 (MDM2), which was detected using the Co-immunoprecipitation (co-IP) assay and glutathione S-transferase (GST) pull-down assay. The expression of proteins in MDM2/p53 signaling was detected by western blot analysis. Results indicated that TRIM47 expression was highly expressed in PCa cells. TRIM47 knockdown inhibited PCa proliferation and cell cycle whereas promoted cell apoptosis. Besides, TRIM47 knockdown significantly inhibited the migration and invasion of PCa cells. In addition, TRIM47 was proved to bind to MDM2 and regulated MDM2/p53 expression. Importantly, MDM2 overexpression counteracted the impacts of TRIM47 knockdown on cell viability, cell cycle, apoptosis, migration and invasion by regulating the MDM2/p53 pathway. Collectively, our results suggested that TRIM47 silencing inhibits the malignant biological behaviors of prostate cancer cells by regulating MDM2/p53 signaling, which may provide a novel therapeutic target for PCa treatment.

Controllable Fabrication of Sub-10 nm Graphene Nanopores via Helium Ion Microscopy and DNA Detection.

Solid-state nanopores have become a prominent tool in the field of single-molecule detection. Conventional solid-state nanopores are thick, which affects the spatial resolution of the detection results. Graphene is the thinnest 2D material and has the highest spatial detection resolution. In this study, a graphene membrane chip was fabricated by combining a MEMS process with a 2D material wet transfer process. Raman spectroscopy was used to assess the quality of graphene after the transfer. The mechanism behind the influence of the processing dose and residence time of the helium ion beam on the processed pore size was investigated. Subsequently, graphene nanopores with diameters less than 10 nm were fabricated via helium ion microscopy. DNA was detected using a 5.8 nm graphene nanopore chip, and the appearance of double-peak signals on the surface of 20 mer DNA was successfully detected. These results serve as a valuable reference for nanopore fabrication using 2D material for DNA analysis.

Development and characterization of a bacterial enzyme cascade reaction system for efficient and stable PET degradation.

The widespread use of polyethylene terephthalate (PET) in various industries has led to a surge in microplastics (MPs) pollution, posing a significant threat to ecosystems and human health. To address this, we have developed a bacterial enzyme cascade reaction system (BECRS) that focuses on the efficient degradation of PET. This system harnesses the Escherichia coli (E. coli) surface to display CsgA protein, which forms curli fibers, along with the carbohydrate-binding module 3 (CBM3) and PETases, to enhance the adsorption and degradation of PET. The study demonstrated that the BECRS achieved a notable PET film degradation rate of 3437 ± 148 µg/(d*cm²), with a degradation efficiency of 21.40% for crystalline PET MPs, and the degradation products were all converted to TPA. The stability of the system was evidenced by retaining over 80% of its original activity after multiple uses and during one month of storage. Molecular dynamics simulations confirmed that the presence of CsgA did not interfere with the enzymatic activity of PETases. This BECRS represents a significant step forward in the biodegradation of PET, particularly microplastics, offering a practical and sustainable solution for environmental pollution control.

Biomechanical comparison of a new undercut thread design vs. the V-shape thread design for pedicle screws.

BACKGROUND CONTEXT: Thread shape is regarded as an important factor influencing the fixation strength and osseointegration of bone screws. However, commercial pedicle screws with a V-shaped thread are prone to generating stress concentration at the bone-screw interface, thereby increasing the risk of screw loosening. Thus, modification of the pedicle-screw thread is imperative. PURPOSE: This study aimed to investigate the fixation stability of pedicle screws with the new undercut thread design in comparison to pedicle screws with a V-shaped thread. STUDY DESIGN: In vitro cadaveric biomechanical test and finite element analysis (FEA). METHODS: Pedicle screws with the undercut thread (characterized by a flat crest feature and a tip-facing undercut feature) were custom-manufactured, whereas those with the V-shaped thread were procured from a commercial supplier. Fixation stability was assessed by the cyclic nonpullout compressive biomechanical testing on cadaveric female osteoporotic vertebrae. The vertical displacement and rotation angle of the 2 types of pedicle screws were calculated every 100 cycles to evaluate their resistance to migration and rotation. FEA was conducted to investigate the stress distribution and bone damage at the bone-screw interface for both types of pedicle screws. RESULTS: Biomechanical testing revealed that the pedicle screws with the undercut thread exhibited significantly lower vertical displacement and rotation angles than the pedicle screws with the V-shape thread (P < 0.05). FEA results demonstrated a more uniform stress distribution in the bone surrounding the thread in the undercut design than in the V-shape design. Additionally, bone damage resulting from the pedicle screw was lower in the undercut design than in the V-shape design. CONCLUSIONS: Pedicle screws with an undercut thread are less prone to migration and rotation and thus more stable in the bone than those with a V-shape thread. CLINICAL SIGNIFICANCE: The undercut thread design may reduce the incidence of pedicle-screw loosening.

Research on prediction of coal water medium separation effect based on multi-models.

To improve the separation efficiency of raw coal and ensure clean use, the accurate calculation of the partition coefficients (PCs) in coal water medium sorting is required. Single models have been used to predict the partition coefficient (PC) for decades, but their accuracy remains constrained. This study proposes a multi-model (MM) calculation method based on the Gompertz model (GM), the Logistic model (LM), the Arctangent model (AM), and the Approximate formula (AFM) to improve the accuracy of the predicted coal water medium sorting PCs. Four groups of coal samples and two specific cases were used to verify the accuracy of the MM calculation method. The PCs of the MM method had a minimal Ef (0.91-8.84), a maximal R2 (0.9648-0.9994), a maximal F-value (199.17-11352.31), and the highest significance of all the models. The MM method was found to be the most suitable of all the models for predicting any coal water medium separation process. Further, when calculating the PC for cleaned coal ash, the separation density of MM is closer to the actual separation density than that of either the GM, LM, AM, or AFM models. The MM method, therefore, produces more accurate results compared to a single model. MM is expected to predict the PC based on the required cleaned coal ash, and then regulate the sorting density to improve the production efficiency.

Investigating the Mechanism of Diethylenetriamine Pentamethylene Phosphonic Acid as a Depressant for Calcite Flotation of Fluorite.

Fluorite and calcite have been attracting research attention for a long time. This paper reports on an investigation of the use of diethylene triamine pentamethylphosphonic acid (DTPMPA) as a chelating inhibitor. DTPMPA was used as a chelating inhibitor to study the flotation, separation, and adsorption behaviors of fluorite and calcite minerals. The microflotation experiment showed that the maximum separation of fluorite and calcite can be achieved with a DTPMPA dosage of 1.5 × 10-4 mol/L under weakly alkaline conditions (pH = 8). Zeta potential measurement, Fourier transform infrared spectroscopy, and X-ray photoelectron spectroscopy were used to confirm that DTPMPA was adsorbed on the surface of calcite, inhibiting NaOl adsorption. Additionally, density functional theory calculations showed that oxygen in the DTPMPA phosphate group formed the most stable bidentate binuclear adsorption configuration by chelating with calcium on the calcite surface. Through detection analysis and simulation calculations, the results showed that DTPMPA exhibited significantly weaker adsorption on fluorite compared to that on calcite, highlighting its selective inhibition ability on calcite.

Three-dimensional comprehensive evaluation of unilateral alveolar cleft bone grafting with iliac bone and chin bone blocks.

In this study, we aimed to provide guidance for selecting bone grafting materials in cases of alveolar clefts. Twenty-nine patients with unilateral complete alveolar clefts were categorized into three groups based on the bone grafting material used: Group A (iliac bone block grafts), Group B (iliac cancellous bone grafts), and Group C (chin bone block grafts). Cone-beam computed tomography (CBCT) data were analyzed using Mimics 19.0 software. Results showed that Group A had the highest bone formation rate, with significant differences observed between Groups A and B, as well as between Groups B and C. Group A and Group C had the highest proportion of Type I in volume assessment, while Group B had the highest proportion of Type III, Significant differences were observed in the distribution of volume assessment scores among the three groups. Bone height measurement results indicated that buccal-side measurement points had a higher proportion of Type I bone height than palatal-side measurement points. Bone width measurement results showed that Type I bone width was highest in Group C, while Type IV bone width was highest in Group B. Significant differences were observed in the distribution of implanted bone width among the three groups. Total grafting scores indicated that Types A and D were predominant in Groups A and C, while Group B had the highest proportion of Type D. Significant differences were observed in the distribution of total grafting scores among the three groups. The comprehensive evaluation method provides accurate assessment of alveolar cleft bone grafting outcomes and is applicable in clinical settings. Based on the results, we consider both iliac bone blocks and chin bone blocks as suitable materials for alveolar cleft bone grafting. Grafting material selection should consider preoperative gap volume measured using CBCT, required bone quantity, donor site complications, and overall clinical needs.

Mechanical properties and tissue damage caused by staples in gastrointestinal anastomosis.

Gastrointestinal surgery using a stapler is usually associated with tissue damage, anastomosis leakage, bleeding, and other complications, which is one of the effective methods for treating digestive tract cancer. The cutting properties of staples and the tissue damage occurring in the process of stapling porcine esophageal and gastric tissues have been evaluated and a new type of stapler has been designed. Since different structural and mechanical properties esophageal and gastric tissues layers, the puncturing force exhibits a fluctuating trend. Compressive stress caused by the bending of the staple legs can lead to the destruction of the vascular network inside the tissue, tissue deforms and tears. Finally, a staple with an internal incision arc (IIA) tip is designed, which meeting the performance requirements.

Chemical Structure Characteristics and Model Construction of Coal with Three Kinds of Coalification Degrees.

The coal structure is extensively used for studying the properties of coal, and the construction of accurate and reliable coal structure models can promote these researches. In this study, Fourier transform infrared (FTIR) spectroscopy and X-ray diffraction (XRD) were used to analyze the changes in the coal structure as a function of the coalification degree, and a semiquantitative model construction method based on FTIR, XRD, and X-ray photoelectron spectroscopy (XPS) analyses was proposed. With an increase in the coalification degree, the size of the aromatic cores in the coal increased. During the conversion from lignite to bituminous coal, the content of aliphatic structures increased, while the content of oxygen-containing functional groups (OFGs) significantly decreased. Conversely, during the conversion from bituminous coal to anthracite, the content of aliphatic structures decreased while the content of OFGs slightly increased. The calculated FTIR spectra of the coal structure models were consistent with the experimental FTIR spectra, which confirmed the accuracy of the models. Furthermore, the models successfully explained the microscopic mechanism underlying the differences in the wettability of the coal samples with varying coalification degrees. The construction method and coal structure models in this study will be more widely applied in future research.