Enamel matrix derivative expedites osteogenic differentiation of BMSCs via Wnt/ß-catenin pathway in high glucose microenvironment.

INTRODUCTION: The influence of enamel matrix derivative (EMD) on proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) was explored in high glucose (HG) microenvironment with interaction of Wnt/ß-catenin pathway. MATERIALS AND METHODS: Extraction of BMSCs from Sprague-Dawley rats, culture, and identification were manifested. The cells were treated with different concentration of EMD in HG to figure out the most available concentration for proliferation and osteogenic differentiation. Then, observation of cell growth curve and cell cycle changes, and detection of Osterix, runt-related transcription factor 2 (Runx2), COL-I, early osteogenic indexes, Calcium salt deposition, and ß-catenin protein in Wnt/ß-catenin pathway were assured. After adding Wnt/ß-catenin pathway inhibitor (XAV-939) in the cells with osteogenesis induction, detection of binding of ß-catenin to Osterix was clarified. RESULTS: Via identification BMSCs cultured in vitro was qualified. Different concentrations of EMD could accelerate cell proliferation in HG and osteogenesis induction, and 75 µg/mL EMD had the best effect. The HG augmented BMSCs proliferation and the propidium iodide index of flow cytometry cycle was elevated in HG, which were strengthened via the EMD. After BMSCs' osteogenesis induction, Osterix, Runx2, CoL-1, early osteogenic indexes, and calcium salt deposition were reduced, but elevated via EMD. ß-Catenin was the lowest in the HG, but elevated after EMD. After addition of XAV-939, reduction of ß-catenin and the downstream (Osterix and Runx2) were manifested. Detection of binding protein bands was in ß-catenin and Osterix of the HG after EMD treatment. CONCLUSION: EMD may facilitate the osteogenic differentiation of BMSCs via activating the Wnt/ß-catenin pathway in HG.

Constitutive activity of a G protein-coupled receptor, DRD1, contributes to human cerebral organoid formation.

The intricate balance of neural stem cell (NSC) amplification and neurogenesis is central to nervous system development. Dopamine D1 receptor (DRD1) is a typical G protein-coupled receptor (GPCR) mainly expressed in neurogenic area, with high constitutive activity. The receptor appears in the embryonic period before the formation of mature synaptic contacts, which indicates that dopamine receptor and its constitutive activity play crucial roles in the embryonic brain development. Here, we found that DRD1 was enriched in human NSCs. Inhibition of the receptor activity by its inverse agonists promoted human NSCs proliferation and impeded its differentiation. These results were also mimicked by genetic knockdown of DRD1, which also blocked the effects of inverse agonists, suggesting a receptor-dependent manner. More interestingly, knock-in A229T mutant with reduced DRD1 constitutive activity by CRISPR-Cas9 genome editing technology resulted into increased endogenous human NSCs proliferation. These results were well reproduced in human cerebral organoids, and inhibition of the DRD1 constitutive activity by its inverse agonists induced the expansion and folding of human cerebral organoids. The anatomic analysis uncovered that decreasing the constitutive activity of DRD1 by its inverse agonists promoted the NSCs proliferation and maintenance that led to hindered cortical neurogenesis. Further mechanistic studies revealed that the PKC-CBP pathway was involved in the regulation by DRD1. Thus, our findings indicate that the constitutive activity of DRD1 and possibly other GPCRs plays an important role in the development of human nervous system.

The Systemic Immune-Inflammation Index and Albumin as Prognostic Predictors in Laryngeal Carcinoma.

BACKGROUND: Systemic inflammation via host-tumor interactions is currently recognized as the seventh cancer hallmark. The purpose of this study was to detect whether pretreatment peripheral indexes were associated with aggressive behavior and prognosis of laryngeal carcinoma patients. METHODS: The pretreatment peripheral indexes such as albumin and systematic immune-inflammation index (SII) in 338 patients with laryngeal carcinoma were retrospectively recorded, the relationships between them and clinicopathological features and prognosis were analyzed. RESULTS: A high SII value was significantly positively associated with age (P = 0.01), N stage (P = 0.022) and tumor differentiation (P = 0.001). A low albumin value was significantly negatively associated with age (P = 0.01), tumor location (P = 0.001) and T stage (P = 0.015), N stage (P = 0.001) and tumor differentiation (P = 0.001). Univariate and multivariate survival analysis showed that a high SII (HR: 2.415, 95% CI 1.400-4.184; P = 0.002), a low blood albumin content (HR: 3.194, 95% CI 2.030-5.025; P = 0.001) independently predicted poor overall survival (OS). However, neutrophil-lymphocyte ratio (NLR), platelet-lymphocyte ratio (PLR) and platelet distribution width (RDW) were not independent prognostic factors. CONCLUSION: Pretreatment peripheral indexes SII and albumin could function as inexpensive indicators of aggressive behavior and be feasible and promising predictive biomarkers for prognosis in laryngeal carcinoma patients. Quantification of pretreatment SII and albumin may help physicians to design more effective management and follow-up strategies in laryngeal carcinoma patients.

LAMC2 acts as a novel therapeutic target of cetuximab in laryngeal cancer.

This study was set out to determine the function of LAMC2 in laryngeal cancer (LC). Initially, we identified the expression of LAMC2 in LC cells and tissues using TCGA datasets, GEO datasets (GSE143224), qRT-PCR, and western blot. Besides, we analyzed the correlations between LAMC2 and clinicopathologic features in LC patients. The CCK-8 assays were performed to detect cell viability and the half-maximal inhibitory concentration of cetuximab (IC50) in LC cells. We explored the correlations between LAMC2 and EGFR and further explored the regulation mechanism of cetuximab in LC. This study identified a high expression of LAMC2 in LC cells and tissues. The expression levels of LAMC2 were associated with TNM classification, lymph node (LN) metastasis, differentiation, and overall survival (OS). LAMC2 significantly promoted cell proliferation and cell viability. Besides, cetuximab significantly inhibited LAMC2 expression levels. LAMC2 significantly reversed the effect of cetuximab suppressing cell proliferation in LC cells. In conclusion, LAMC2 may act as a novel anti-cancer target in LC.

Design and evaluation of a zero-order controlled release system based on pre-hydrated constant release area prepared by compression coating technology.

The main aim of this research work was to develop and evaluate a drug delivery system with compression coating technology to control drug release at a constant rate. The compression coated tablets (CCTs) consist of the hydrophilic matrix core and the hydrophobic waxy coating. The presence of hydrophobic waxy coating could provide sufficient time for hydration of the core to prevent initial burst release. The mechanism research revealed that erosion was the main way of drug release and the releasing area was constant during the entire release process because the core tablet was located in the cup-shaped coating after one side cover was dropped at the lag time. This made the release behavior exhibit zero-order kinetics (R2>0.99). The coating rupture strength and the core swelling force at the lag time influenced erosion rate thus affecting release rate. Different solubility of drugs (propranolol hydrochloride, melatonin, and nifedipine) was selected as model drugs and the properties of the prepared CCTs in terms of formulations and in vitro release were evaluated. The release rate was independent of solubility, medium pH, and osmotic pressure. This zero-order controlled system could be applied to both controlled drug delivery and chrono pharmaceutical drug delivery.

Identification of Arylphorin interacting with the insecticidal protein PirAB from Xenorhabdus nematophila by yeast two-hybrid system.

PirAB toxin was initially found in the Photorhabdus luminescens TT01 strain and is a demonstrated binary toxin with high insecticidal activity. In this paper, we co-expressed the pirAB gene of Xenorhabdus nematophila HB310 in a prokaryotic expression system, and we found that the PirAB protein showed high hemocoel insecticidal activity against Galleria mellonella, Helicoverpa armigera and Spodoptera exigua. LD50 values were 1.562, 2.003 and 2.17 µg/larvae for G. mellonella, H. armigera, and S. exigua, respectively (p > 0.05). Additionally, PirAB-interaction proteins were identified from G. mellonella by 6 × His Protein Pulldown combined with liquid chromatography-tandem mass spectrometry (LC-MS/MS). Of which, arylphorin of G. mellonella showed the highest matching rate. A protein domain conservative structure analysis indicated that arylphorin has three domains including Hemocyanin-N, Hemocyanin-M, and Hemocyanin-C. Among these protein domains, Hemocyanin-C has immune and recognition functions. Further, Hemocyanin-C domain of arylphorin was identified to interact with PirA but not PirB by Yeast two-hybrid system. These findings reveal, for the first time, new host protein interacting with PirAB. The identification of interaction protein may serve as the foundation for further study on the function and insecticidal mechanism of this binary toxin from Xenorhabdus.

NF-κB directly regulates ß-arrestin-1 expression and forms a negative feedback circuit in TNF-α-induced cell death.

ß-Arrestins (ß-arrestin-1 and -2) are multifunctional proteins that play important roles in the regulation of inflammation and cell survival that need to be tightly controlled; however, the mechanism that underlies their gene expression is largely unclear. Here, we demonstrate that ß-arrestin-1 is a transcriptional target of NF-κB. mRNA and protein levels of ß-arrestin-1 were up-regulated by NF-κB inducers. Inhibition of NF-κB prevented the up-regulation of ß-arrestin-1 mRNA, whereas activation of NF-κB led to increased ß-arrestin-1 expression. ß-Arrestin-1 promoter activity was consistently enhanced upon NF-κB activation as a result of the presence of a highly conserved κB site. ß-Arrestin-1, in turn, suppressed the transcriptional activity of NF-κB by interfering with the interaction between p65 and p50. ß-Arrestin-1-deficient mice displayed reduced TNF-α-induced cell death and increased expression of antiapoptotic genes. Reintroduction of ß-arrestin-1, but not its mutant, which is unable to interfere with the p65-p50 interaction, into ß-arrestin-deficient mouse embryonic fibroblasts partially restored sensitivity to TNF-α-induced cell death. These findings reveal NF-κB and ß-arrestin-1 to be key components of a negative feedback circuit that is necessary to regulate cell death.-Li, J., Guo, A., Wang, Q., Li, Y., Zhao, J., Lu, J., Pei, G. NF-κB directly regulates ß-arrestin-1 expression and forms a negative feedback circuit in TNF-α-induced cell death.

Expression and characterization of two chitinases with synergistic effect and antifungal activity from Xenorhabdus nematophila.

Xenorhabdus nematophila HB310 secreted the insecticidal protein toxin complex. Two chitinase genes, chi60 and chi70, were found in X. nematophila toxin complex locus. In order to clarify the function of two chitinases, chi60 and chi70 genes were cloned and expressed in Escherichia coli Transetta (DE3). As a result, we found that the Chi60 and Chi70 belonged to glycoside hydrolases (GH) family 18 with a molecular mass of 65 kDa and 78 kDa, respectively. When colloidal chitin was treated as the substrate, Chi60 and Chi70 were proved to have the highest enzymatic activity at pH 6.0 and 50 °C. Chi60 and Chi70 had obvious growth inhibition effect against the second larvae of Helicoverpa armigera with growth inhibiting rate of 81.99% and 90.51%. Chi70 had synergistic effect with the insecticidal toxicity of Bt Cry 1Ac, but the Chi60 had no synergistic effect with Bt Cry 1Ac. Chi60 and Chi70 showed antifungal activity against Alternaria brassicicola, Verticillium dahliae and Coniothyrium diplodiella. The results increased our understanding of the chitinases produced by X. nematophila and laid a foundation for further studies on the mechanism of the chitinases.

Expression of Neu5Acα2,3Gal and Neu5Acα2,6Gal on the nasal mucosa of patients with chronic rhinosinusitis and its possible effect on bacterial biofilm formation.

OBJECTIVE: Adherence of pathogen to nasal mucosa and colonization is the first step of bacterial biofilm(BBF) formation in patients with chronic rhinosinusitis (CRS).Terminal sialic acids presenting on cell surface are potential targets for bacterial binding, thus may partly contribute to the pathogenesis of CRS. However, little has been published in this respect, the purpose of our study aimed to investigate the expression of sialic acids on the nasal mucosa in CRS patients and its possible effect on BBF formation. METHODS: Sinus mucosa were harvested from CRS patients undergoing endoscopic surgery. The positive of BBF formation were detected by scanning electronic microscopy (SEM) and the expression of Neu5Acα2,3Gal(α2,3-linked sialic acid) and Neu5Acα2,6Gal(α2,6-linked sialic acid) on nasal mucosa were determined by fluorescent-immunohistochemical staining (F-IHC) with MAL-II and SNA respectively. A semi-quantitative scoring system was used to assess their different expression between CRS group and the control, as well as BBF positive and negative group. RESULTS: Expression of Neu5Acα2,3Gal and Neu5Acα2,6Gal were both detected in the epithelium and submucosal glands of all 40 CRS patients and 23 controls, they were significantly up-regulated in CRS group(p < 0.05). Among 24 CRS patients, typical BBF formation were identified in 13 cases while the other 11 were regarded as negative, Between the subgroup of BBF(+) and BBF(-), both of Neu5Acα2,3Gal and Neu5Acα2,6Gal had a trend of increasing in BBF(+) group, however, the increased expression of Neu5Acα2,3Gal was statistical significance (4.77 ± 0.90 versus 3.45 ± 1.40; p = 0.0282), whereas the difference of Neu5Acα2,6Gal was insignificant(4.15 ± 1.27 versus 3.55 ± 1.59; p = 0.4281). CONCLUSION: Expression of MAL-II binding (most probable Neu5Acα2,3Gal) and SNA binding (Neu5Acα2,6Gal) were up-regulated in inflamed nasal mucosa, and the increased expression of them may contribute to bacterial biofilm formation which deserved a further investigation.

Aminopeptidase N5 (APN5) as a Putative Functional Receptor of Cry1Ac Toxin in the Larvae of Athetis lepigone.

Athetis lepigone was a new lepidopteran pest and caused severe damage to maize crops in China. We have detected that Cry1Ac protoxin and toxin were highly active against the larvae of A. lepigone. However, there is no report about the mode of action of Bt Cry1Ac toxin against this pest until now. A 110 kDa APN5 protein from BBMV of A. lepigone was identified as the binding receptor of Cry1Ac toxin using Ligand blotting. The Cry1Ac receptor APN5 was cloned from A. lepigone larval midgut mRNA and named as AlAPN5 (GenBank accession no.: KU950745). AlAPN5 had a GATEN motif and been classified to Class 5 APNs. 79.2% reduction in mortality was observed when A. lepigone larvae were injected with siRNA of the AlAPN5 gene and treated with Cry1Ac toxin. These data demonstrate that AlAPN5 is a putative functional receptor and maybe the only receptor of Cry1Ac in A. lepigone.

PirAB protein from Xenorhabdus nematophila HB310 exhibits a binary toxin with insecticidal activity and cytotoxicity in Galleria mellonella.

PirAB (Photorhabdus insect-related proteins, PirAB) toxin was initially found in the Photorhabdus luminescens TT01 strain and has been shown to be a binary toxin with high insecticidal activity. Based on GenBank data, this gene was also found in the Xenorhabdus nematophila genome sequence. The predicted amino acid sequence of pirA and pirB in the genome of X. nematophila showed 51% and 50% identity with those gene sequences from P. luminescens. The purpose of this experiment is to identify the relevant information for this toxin gene in X. nematophila. The pirA, pirB and pirAB genes of X. nematophila HB310 were cloned and expressed in Escherichia coli BL21 (DE3) using the pET-28a vector. A PirAB-fusion protein (PirAB-F) was constructed by linking the pirA and pirB genes with the flexible linker (Gly)4 DNA encoding sequence and then efficiently expressed in E. coli. The hemocoel and oral insecticidal activities of the recombinant proteins were analyzed against the larvae of Galleria mellonella. The results show that PirA/B alone, PirA/B mixture, co-expressed PirAB protein, and PirAB-F all had no oral insecticidal activity against the second-instar larvae of G. mellonella. Only PirA/B mixture and co-expressed PirAB protein had hemocoel insecticidal activity against G. mellonella fifth-instar larvae, with an LD50 of 2.718µg/larva or 1.566µg/larva, respectively. Therefore, we confirmed that PirAB protein of X. nematophila HB310 is a binary insecticidal toxin. The successful expression and purification of PirAB laid a foundation for further studies on the function, insecticidal mechanism and expression regulation of the binary toxin.

Management of maxillary sinus inverted papilloma via endoscopic partial medial maxillectomy with an inferior turbinate reversing approach.

The aim of this study is to evaluate the efficacy of endoscopic treatment for maxillary inverted papilloma (IP) through partial medial maxillectomy with an inferior turbinate reversing approach. A retrospective analysis of patients treated in our institution for maxillary sinus IP between July 2011 and August 2015 was performed. Demographics, operative technique, characteristics of tumors, complications, postoperative follow-up, and recurrence were evaluated. Twenty-two patients were enrolled in the study. All tumor attachments were identified intraoperatively. Adequate visualization was obtained following our approach. All inferior turbinate and nasolacrimal ducts were preserved. The median follow-up time was 41 months. One recurrence occurred at the follow-up time of 27 months. Postoperative hemorrhage and numbness at the ipsilateral frontal teeth were reported in two and one patients, respectively. Endoscopic surgery through partial medial maxillectomy using an inferior turbinate reversing approach provides full access to the maxillary sinus and preserves the inferior turbinate and nasolacrimal duct.

[Associations between hepatitis B virus x gene mutations and hepatocellular carcinoma].

OBJECTIVE: To investigate the association of mutations in the hepatitis B virus (HBV) X gene (HBX, encoding the HBx protein) and development of hepatocellular carcinoma (HCC). METHODS: Forty-four patients with HBV-related HCC participated in the study, along with 76 patients with chronic HBV infection who assessed as controls. All patients had serum HBV DNA levels that were higher than 10(3) copies/ml. Extracted HBV DNA was subjected to nested PCR to amplify the HBX gene, followed by direct sequencing. All sequencing data were compared to the consensus HBV sequence to identify mutations. The sequencing data were analyzed by Chromas and SeqMan software. RESULTS: Mutations of G1467C, G/C1479A, C1485T and C1653T in the X region were found, but did not show any significant difference in occurrence between the HCC group and the chronic HBV infection group (P>0.05). The T1674C mutation in the X region, however, occurred more frequently in the HCC group (29.27% vs.6.67%, P<0.05). Prevalence of the T1753C mutation and the A1762T/G1764A double mutation in the BCP region was significantly higher in the HCC group than in the chronic HBV infection group (P<0.05) and in the group of patients with hepatitis B e antigen (HBeAg)-negative status compared to the patients with HBeAg-positive status (P<0.05). CONCLUSION: Incidence of the T1674C mutation in the X region and of the T1753C mutation and the A1762T/G1764A double mutation in the BCP region was higher for patients with HBV-related HCC; the T1753C mutation and the A1762T/G1764A double mutation may inhibit the formation of HBeAg.

Detection of minimum activity fragment of a novel Cry7Ab3 protein specific to Henosepilachna vigintioctomaculata (Coleopteran: Coccinellidae).

Bacillus thuringiensis Cry7Ab3 toxin has insecticidal activity against larvae of Henosepilachna vigintioctomaculata. Cry7Ab3 toxin is solubilized under alkaline condition and activated by proteases within the larval gut. In order to assess the functions of the N- and C-terminal regions, several N- and C-terminal truncated forms of Cry7Ab3 were constructed. It was determined that amino acid removal at the N-terminal, which disrupt the α-helico structure, resulted in the inactivation of the protein. The deletion of 512 amino acids from the C-terminus reduced the toxicity. However, the deletion of 481 amino acids from the C-terminus resulted in the highest activity. These findings directly demonstrated the critical roles of N- and C-terminal amino acids on the toxicity of Cry7Ab3.

Laryngo-tracheobronchial amyloidosis: a case report and review of literature.

AMYLOIDOSIS is a benign process which can have systemic involvement. Though larynx is the common site of localized amyloidosis in the head and neck region,1 it was seldom reported with heterochronous implication of bilateral ventricles. Here we report a case of laryngeal amyloidosis heterochronously localized at bilateral ventricles with tracheobronchial involvement. Combined with our experience we reviewed the literature, and discuss the pertinent managements of this condition.

Organoids in gastrointestinal diseases: from bench to clinic.

The etiology of gastrointestinal (GI) diseases is intricate and multifactorial, encompassing complex interactions between genetic predisposition and gut microbiota. The cell fate change, immune function regulation, and microenvironment composition in diseased tissues are governed by microorganisms and mutated genes either independently or through synergistic interactions. A comprehensive understanding of GI disease etiology is imperative for developing precise prevention and treatment strategies. However, the existing models used for studying the microenvironment in GI diseases-whether cancer cell lines or mouse models-exhibit significant limitations, which leads to the prosperity of organoids models. This review first describes the development history of organoids models, followed by a detailed demonstration of organoids application from bench to clinic. As for bench utilization, we present a layer-by-layer elucidation of organoid simulation on host-microbial interactions, as well as the application in molecular mechanism analysis. As for clinical adhibition, we provide a generalized interpretation of organoid application in GI disease simulation from inflammatory disorders to malignancy diseases, as well as in GI disease treatment including drug screening, immunotherapy, and microbial-targeting and screening treatment. This review draws a comprehensive and systematical depiction of organoids models, providing a novel insight into the utilization of organoids models from bench to clinic.

Microbial and metabolic profiles unveil mutualistic microbe-microbe interaction in obesity-related colorectal cancer.

Obesity is a risk factor for colorectal cancer (CRC), and the involvement of gut microbiota in the pathogenesis of obesity and CRC is widely recognized. However, the landscape of fecal microbiome and metabolome distinguishing patients with obesity-related CRC from obesity remains unknown. Here, we utilize metagenomic sequencing and metabolomics from 522 patients with CRC and healthy controls to identify the characteristics of obese CRC. Our integrated analysis reveals that obesity-related CRC is characterized by elevated Peptostreptococcus stomatis, dysregulated fatty acids and phospholipids, and altered Kyoto Encyclopedia of Genes and Genomes pathways involving glycerophospholipid metabolism and lipopolysaccharide synthesis. Correlation analysis unveils microbial interactions in obesity, where the probiotic Faecalibacterium prausnitzii and the tumor-promoting species P. stomatis may engage in cross-feeding, thereby promoting tumorigenesis. In vitro experiments affirm enhanced growth under cross-feeding conditions. The mutualistic microbe-microbe interaction may contribute to the association between obesity and elevated CRC risk. Additionally, diagnostic models incorporating BMI-specific microbial biomarkers display promise for precise CRC screening.

Anticancer mechanism of apigenin and the implications of GLUT-1 expression in head and neck cancers.

Apigenin, a natural phytoestrogen flavonoid, has potential biological effects, including antioxidative, anti-inflammatory and anticancer activities. The mechanisms of anticancer activities of apigenin are unknown. Some studies have found that apigenin inhibits GLUT-1 mRNA and protein expression in cancer cells. Thus, we hypothesized that apigenin exerts similar effects on head and neck cancers through its inhibition of GLUT-1 expression. In this article, we review the anticancer mechanism of apigenin and the implications of GLUT-1 expression in head and neck cancers. In addition, we describe the current state of knowledge about the relationship between apigenin and GLUT-1 expression in head and neck cancers.

Effect of antisense oligodeoxynucleotides glucose transporter-1 on enhancement of radiosensitivity of laryngeal carcinoma.

PURPOSE: Laryngeal carcinomas always resist to radiotherapy. Hypoxia is an important factor in radioresistance of laryngeal carcinoma. Glucose transporter-1 (GLUT-1) is considered to be a possible intrinsic marker of hypoxia in malignant tumors. We speculated that the inhibition of GLUT-1 expression might improve the radiosensitivity of laryngeal carcinoma. METHODS: We assessed the effect of GLUT-1 expression on radioresistance of laryngeal carcinoma and the effect of GLUT-1 expressions by antisense oligodeoxynucleotides (AS-ODNs) on the radiosensitivity of laryngeal carcinoma in vitro and in vivo. RESULTS: After transfection of GLUT-1 AS-ODNs: MTS assay showed the survival rates of radiation groups were reduced with the prolongation of culture time (p<0.05); Cell survival rates were significantly reduced along with the increasing of radiation dose (p<0.05). There was significant difference in the expression of GLUT-1mRNA and protein in the same X-ray dose between before and after X-ray radiation (p<0.05). In vivo, the expressions of GLUT-1 mRNA and protein after 8Gy radiation plus transfection of GLUT-1 AS-ODNs were significant decreased compared to 8Gy radiation alone (p<0.001). CONCLUSION: Radioresistance of laryngeal carcinoma may be associated with increased expression of GLUT-1 mRNA and protein. GLUT-1 AS-ODNs may enhance the radiosensitivity of laryngeal carcinoma mainly by inhibiting the expression of GLUT-1.

Expression of hypoxia-related markers in inflammatory myofibroblastic tumors of the head and neck.

BACKGROUND: The etiology of inflammatory myofibroblastic tumors (IMTs) is controversial and the prognosis is unpredictable. Previous studies have not investigated the expression of hypoxia-related markers in IMTs. METHODS: Between 2002 and 2012, 12 consecutive patients with histologically proven IMTs were enrolled in the study. Immunohistochemistry was used to detect GLUT-1, HIF-1α, PI3K, and p-Akt expression in paraffin-embedded tumor specimens. Associations among GLUT-1, HIF-1α, PI3K, and p-Akt protein expression and clinical parameters were investigated. RESULTS: The mean duration of follow-up was 52.1 months (range, 11 to 132 months). Six patients had local recurrence. GLUT-1, HIF-1α, PI3K, and p-Akt expression were detected in 41.7%, 50.0%, 33.3%, and 41.7% of patients, respectively. Fisher's exact test revealed significant correlations between recurrence of IMT and PI3K expression (P = 0.01) and p-Akt expression (P = 0.015). Univariate analyses revealed significant correlations between survival and GLUT-1 expression (P = 0.028), PI3K expression (P = 0.006), and p-Akt expression (P = 0.028). Multivariate analysis did not show a significant relationship between survival and GLUT-1, HIF-1α, PI3K, or p-Akt. Spearman rank correlation analysis showed significant correlations between HIF-1α and PI3K expression (r = 0.707, P = 0.01) and between p-Akt and PI3K expression (r = 0.837, P = 0.001). CONCLUSIONS: Although our results are inconclusive owing to the small sample size, they suggest that PI3K and p-Akt expression may play a role in the recurrence of IMTs of the head and neck.