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Laryngeal squamous cell carcinoma (LSCC) is a laryngeal malignancy with a high mortality rates, and its treatment remains difficult. Sevoflurane is a surgical anesthesia which has anti-tumor effect. This investigation assessed the effects of LSCC cells treatment with Sevoflurane in vitro and in vivo. Hep-2 and Tu177 cells, human LSCC samples and BALB/C nude mice were used for result assessments. Cell viability, proliferation, migration and invasion were assessed via Cell Count Kit-8, wound healing assay and transwell invasion assay respectively. MiR-26a and FOXO1 expressions was examined by qRT-PCR. FOXO1, E-cadherin, N-cadherin and vimentin activities were examined by Western blotting. Moreover, animal experiments were performed to verify our findings in vitro. Lastly, miR-26a and FOXO1 expression levels in clinical samples were analyzed. According to the results, Sevoflurane decreased LSCC cells’ viability and even stimulated their apoptosis in vitro and in vivo. Moreover, it could reduce the migration, invasion and EMT. Mechanistically, sevoflurane could downregulate miR-26a expression and that miR-26a could negatively modulate FOXO1 activity. Thus, sevoflurane could increase FOXO1 activity. In the clinical samples, miR-26a expression was significantly upregulated, but FOXO1 was remarkably down-regulated and miR-26a expression in LSCC was linked with better prognosis. In conclusion, MiR-26a is increased and FOXO1 is reduced in human LSCC, Sevoflurane inhibits proliferation and mediates apoptosis of LSCC cells. Further, MiR-26a binds FOXO1 directly, and FOXO1 expression is down-regulated by Sevoflurane. Finally, Sevoflurane triggers LSCC cells apoptosis in vivo. Sevoflurane use to target miR-26a/FOXO1 may be a novel alternative for LSCC therapy.

Programmed death‑ligand 1 (PD‑L1), an immune co‑stimulatory molecule, is expressed on various cancer cells and the surface of immune cells. Its overexpression on tumor cells suppresses the immune response to promote tumor cell immune escape. The present study demonstrated that PD‑L1 was critical in head and neck squamous cell carcinoma (HNSCC) carcinogenesis. Immunohistochemical analysis of HNSCC tissue microarrays revealed that PD‑L1 was overexpressed in tumor tissue, and its expression increased as tumor malignancy progressed (from grade I to IV). Subsequently, the expression of PD‑L1 was knocked down or overexpressed in the HNSCC cell lines Cal‑27 and Fadu. It was demonstrated that PD‑L1 significantly induced HNSCC cell proliferation and colony forming ability. Cell proliferation was also promoted in Cal‑27 cell xenograft BALB/c nude mice. In addition, it was determined by western blotting that the PD‑L1‑mediated increase in HNSCC cell proliferation may have been associated with the activation of mammalian target of rapamycin (mTOR) signaling pathway. Furthermore, mTOR inhibitor (rapamycin) prevented the increase in proliferation. Based on these results, it was concluded that PD‑L1 promoted cell proliferation of HNSCC cells through mTOR signaling, and blocking PD‑L1 may be conducive in HNSCC therapy.

Long non-coding RNAs (LncRNAs) have gained widespread interest and attention as vital regulators in cancer occurrence and development. Nonetheless, the functions and mechanisms of lncRNAs involved in nasopharyngeal carcinoma (NPC) are largely unknown. By analysing the data from GSE61218, we identified a novel lncRNA LINC01515 which is altered in NPC. A series of experiments were performed to examine the exact roles of LINC01515 as well as the molecular mechanisms by which LINC01515 acted in NPC. LINC01515 expression was increased in NPC and that high LINC01515 expression was associated with a worse prognosis. Functionally, depletion of LINC01515 resulted in an inhibition of cell proliferation, migration and invasion, while apoptosis was promoted. Mechanistically, LINC01515 facilitated cell division cycle associated 5 (CDCA5) expression via serving as a sponge for miR-325. And more notably, miR-325 suppressed NPC progression in vitro by targeting CDCA5. Furthermore, the anti-tumor property induced by LINC01515 knockdown was partially reversed due to the overexpression of CDCA5. Taken together, LINC01515 exerted the carcinogenic effect in NPC through regulating miR-325/CDCA5 pathway. Our findings shed light on the possibility of exploiting LINC01515 as a prognostic biomarker or therapeutic target in NPC.

This study aimed in fully investigating the toxicities of ouabain to mouse cochlea and the related cellular environment, and providing an optimal animal model system for cell transplantation in the treatment of auditory neuropathy (AN) and sensorineural hearing loss (SNHL).Different dosages of ouabain were applied to mouse round window. The auditory brainstem responses and distortion product otoacoustic emissions were used to evaluate the cochlear function. The immunohistochemical staining and cochlea surface preparation were performed to detect the spiral ganglion neurons (SGNs), Schwann cells and hair cells.Ouabain at the dosages of 0.5 mM, 1 mM and 3 mM selectively and permanently destroyed SGNs and their functions, while leaving the hair cells relatively intact. Ouabain at 3 mM resulted in the most severe SGNs loss and induced significant loss of Schwann cells started as early as 7 days and with further damages at 14 and 30 days after ouabain exposure.The application of ouabain to mouse round window induces damages of SGNs and Schwann cells in a dose- and time-dependent manner, this study established a reliable and accurate animal model system of AN and SNHL.

Regional metastasis is an important prognostic factor for patients with head and neck squamous cell carcinoma (HNSCC). Neuromedin U (Nmu) is a secreted neuropeptide, named due to its potent uterine contraction‑inducing activity. The aim of the present study was to analyze the significance of Nmu in the regional metastasis of HNSCC. The characteristics of 240 patients recruited from the Department of Otolaryngology Head and Neck Surgery, Renmin Hospital of Wuhan University (Wuhan, China) were summarized retrospectively. The positive rate of neck dissection was analyzed according to the material. The expression levels of Nmu in human tumor samples were analyzed using immunohistochemistry. Subsequently, the expression of Nmu was investigated using a tissue microassay to analyze the association between Nmu protein expression and Tumor Node Metastasis (TNM) status. The positive rate of neck dissection was 51.4% in the study sample. The expression levels of Nmu in primary tumors with regional metastasis were higher, compared with those without metastasis. There was increased protein expression of Nmu in the advanced tumor tissues. The data obtained in the present study demonstrated that the expression of Nmu was correlated with regional metastasis and TNM status. Overexpression of Nmu may be involved in the process of regional metastasis of HNSCC, and may serve as a novel and valuable biomarker for predicting regional metastasis in patients with HNSCC.

The abnormal expression of microRNAs (miRNAs) plays a key role in tumorigenesis. In order to identify potential miRNA biomarkers for early diagnosis of larynx carcinoma, we employed a miRNA microarray technique and applied bioinformatic algorithms to characterize miRNA classifiers in early larynx carcinoma and normal esophageal mucosa tissue samples from 69 patients who were selected retrospectively for this study. We identified 47 miRNAs that were significantly differentially expressed in primary larynx tumor tissues compared to normal tissues using a SAM algorithm. Of these, 30 were up-regulated and 17 down-regulated in early larynx cancer, including hsa-miR-657 , which was overexpressed, and hsa-miR-1287 , which was underexpressed. These two candidate miRNA biomarkers were combined as a single classifier to recognize the biological characteristics in early larynx carcinoma. Real-time quantitative reverse-transcription PCR validated the microarray results in both trial and test samples. The hsa-miR-657-hsa-miR-1287 classifier displayed high sensitivity and specificity for discriminating between early larynx carcinoma and normal mucosa tissues, suggesting they may be suitable as potential predictive biomarkers for the early diagnosis of larynx carcinoma.

Leucine-rich-α-2-glycoprotein 1 (LRG1) is considered as a potential biomarker as it is aberrantly expressed in various malignancies. However, there is limited information regarding its role in head and neck squamous cell carcinoma (HNSCC). In the present study, we aimed to explore the expression pattern of LRG1 in HNSCC and its clinicopathological significance. We first analyzed LRG1 gene expression in HNSCC by investigating data obtained from the Gene Expression Omnibus (GEO) database. The results showed that LRG1 was downregulated in HNSCC tissues and its expression level was negatively related to tumor T and N stages and degree of malignancy. Then, we further tested a tissue microassay and clinical samples, respectively, by immunohistochemical staining and western blotting. Consistently, the results revealed that LRG1 expression was decreased in tumor tissues regardless of the grade of the tumor. Moreover, the protein level of LRG1 showed slight differences among four T stages or three N stages. In addition, there were no significant associations between LRG1 protein expression and other clinicopathological parameters such as gender, age, tumor location and clinical staging. These findings imply that downregulation of the expression of LRG1 is correlated with tumorigenesis but not with the development of HNSCC, indicating the potential clinical value of LRG1 in the early diagnosis of HNSCC.

The development of allergic rhinitis is considered to be determined by the interaction between genetic and environmental factors. Surfactant protein D (SP-D) has been proposed to offer protection against allergenic challenge at various levels in allergic responses. The present study aimed to investigate whether polymorphisms within the SFTPD gene (Met11Thr, Ala160Thr, and Ser270Thr) are associated with allergic rhinitis. Genotyping of SFTPD polymorphisms was performed using the pyrosequencing method. The study population comprised 216 patients with allergic rhinitis and 84 normal controls. The frequency of 11Thr/Thr genotype and Thr allele in the patient group was significantly higher than that in the control group after applying Bonferroni corrections (P = 0.007 and P = 0.006, respectively). Our subjects with the 11Thr/Thr genotype are more susceptible to allergic rhinitis. There were no significant differences between the patient group and the control group for frequencies of genotypes and alleles in either Ala160Thr or Ser270Thr single nucleotide polymorphisms (P > 0.05). No significant associations could be detected between any of these three SFTPD gene polymorphisms and the skin prick test response (P > 0.05). Meanwhile, there was a lack of association between the three loci and the levels of serum total immunoglobulin E (P > 0.05). In summary, our results suggest that the Met11Thr polymorphism in SP-D plays a major role in the genetic predisposition to allergic rhinitis in Chinese adult population, whereas the other two SP-D polymorphisms displayed no significant association with allergic rhinitis.

Objective:Telomerase is an attractive molecular target because it is active in most malignant cells but undetectable in most normal somatic cells. Small, interfering ribonucleic acid segments have been shown to be effective tools for inhibiting the expression of a given gene within human cells. In the present study, we examined the effects of short hairpin ribonucleic acid expression vectors on the growth of laryngeal squamous cell carcinoma in nude mice, and we assessed potential side effects in these animals.Methods:Short hairpin ribonucleic acid expression vectors targeting the messenger ribonucleic acid of the telomerase catalytic unit were constructed and transfected into Hep-2 human laryngeal squamous cells carcinoma in nude mice. Apoptosis and telomerase catalytic unit expression within tumour cells were evaluated after treating with short hairpin ribonucleic acid. Peripheral blood was collected for haematological and biochemical analysis.Results:The findings demonstrated that short hairpin ribonucleic acid plasmids could inhibit tumour cell growth by 76.5 per cent, and that many tumour cells underwent necrotic or apoptotic cell death. There were no significant side effects of short hairpin ribonucleic acid on the heart, liver, kidney, spleen or blood system in this experimental model.Conclusion:These results indicated that the short hairpin ribonucleic acid expression vector targeted at the telomerase catalytic unit of messenger ribonucleic acid significantly inhibited the growth of laryngeal carcinoma in nude mice, with no significant side effects on the experimental animals.

It is well known that crucifers have antitumor effects and 3,3'-diindolylmethane (DIM) is one of the major bioactive components, and the associated molecular mechanisms in a short-term high-dose manner are widely discussed. However, the antitumor effects of DIM in a long-term low-dose manner in nasopharyngeal carcinoma (NPC) has not been reported yet, as to the potential mechanisms in the human body. In the present study, NPC cells were induced by 20 µmol/l DIM for over a month, and the proliferation, apoptosis, migration and in vivo metastasis were investigated. The results showed that DIM significantly reduced the proliferation and migration; however, changes in apoptosis were not observed. In vivo study showed the metastasis was significantly reduced. Compared to the short-term high-dose manner, incomplete similar qualities were observed; next we explored the possible signal pathway revolved, the ERK signaling showed similar changes, while the PI3K/Akt, NF-κB, P38, JNK pathways were significantly altered in the short-term high-dose manner (our previous study) showed no obvious change, indicating the ERK signaling may be the main effector of DIM.

Interleukin–23 (IL–23) is a conventional proinflammatory cytokine that plays a role in tumor progression by inducing inflammation in the tumor microenvironment. However, the role of IL–23 in thyroid cancer migration and invasion remains unclear. In the present study, we observed that the treatment with IL–23, induced migration and invasion in human thyroid cancer cells. Additional data demonstrate that SOCS4 negatively regulates IL-23-mediated migration and invasion. On investigating the mechanisms involved in IL–23 mediated migration and invasion, we observed that miR–25 promotes the migration and invasion of thyroid cancer cells by directly binding to the 3′-UTR of SOCS4 that leads to the inhibition of SOCS4. In addition, we also demonstrated that IL–23 increases miR–25 expression levels, and overexpressed miR–25 is involved in IL-23-associated SOCS4 inhibition and cell migration and invasion. Together, our data suggest that IL–23 induces migration and invasion in thyroid cancer cells by mediating the miR–25/SOCS4 signaling pathway.

To investigate the positive rates of some allergens in different age groups of allergic rhinitis patients, and to analyze the relationship between serum specific immunoglobulin E(sIgE) and total immunoglobulin E(IgE).Six hundred and fifty-three patients with allergic rhinitis, tested by AllergyScreen system, were enrolled in our research.The positive rate of sIgE in all patients was 85.5%. The positive rate of dust mite was the highest with the result of 76.3%, and the positive rate of feline or dogs hair was the lowest of 20.3%. The statistical significance of different positive rate of some allergens were found in the different age groups. All the patients were divided into two groups, one was younger than 18 years old group, which including 18 years old and another was older than 18 years old group. Except Mx, the positive rate of other allergens existed statistical difference. The level of serum total IgE could not reflect the positive rate of sIgE completely.It is an effective and reliable method of the diagnosis of allergic rhinitis to combine the analysis of total serum IgE and sIgE levels with patients clinical manifestations. Different strategies should be employed for the prevention and treatment of allergic rhinitis according to their ages.

Surfactant protein A (SP-A) is protein that appears to play an important role in mammalian first-line host defense. The objective of this study was to immunolocalize SP-A in human sinonasal tissue.Eleven cases of allergic rhinitis, fifteen cases of polyp and seven cases of normal middle turbinate were studied with immunohistochemistry and immunofluorescence method to detect the expression of SP-A.The expression of SP-A in allergic rhinitis and polyp were dramatically higher than that in controls (P0.05), and there was no remarkable difference in the expression of SP-A between allergic rhinitis and polyp (P0.05). The result demonstrated that SP-A was positively correlated with eosinophils within the basement membrane of epithelium (R = 0.81, 0.55). In the result of immunofluorescence, there was significantly higher expression SP-A in nasal mucosa of allergic rhinitis and nasal polyp than that in control group (P0.05).SP-A is likely to play key roles in the inflammatory reaction process of allergic rhinitis and polyp. Its secretion in the upper airway indicates that future studies may allow manipulation of this protein and development of novel treatments for sinonasal pathology.

To analyze the results of serum in allergic rhinitis and investigate the specific clinic allergen and serum immunoglobulin E (IgE) levels.Allergy Screen method was used to detect the specific allergen and total serum IgE level of 134 cases of Allergic rhinitis.The dust mite was the most common allergen in inhalation group in 134 cases of allergic rhinitis, the positive rates was 90%; then were donly, feline and scurfy fungus, the positive rates were 16%, 9%. The positive rates of total IgE was 54%. The serum IgE levels between 100 to 200 kU/L, there was 21 cases together, but there existed 7 negative cases. There were 51 cases' IgE levels more than 200 kU/L, the rates was 70.8%, but there still existed 4 negative cases.Allergy screen method can find relevant allergen and provide basis for the prevention and treatment of allergic disease.

To investigate the expression and clinical significance of RAD51 in laryngocarcinoma.The expression of RAD51 in laryngocarcinoma and polyp of vocal cord tissues were determined by immunohistochemical staining. The results were analyzed and compared with the clinical stage, lymph node metastasis and pathologic grade.(1) The expression of RAD51 in laryngocarcinoma group was extremely stronger than that in polyp of vocal cord group (P0.01). (2) There was significant difference for RAD51 expression in cancer cells between earlier clinical stage group and advanced clinical stage group (P0.01). (3) There was significant difference for RAD51 expression in different pathologic grades (P0.05). (4) There was also significant difference for RAD51 expression between groups with and without lymph node metastasis.RAD51 may play a critical role in tumorigenesis of laryngocarcinoma. RAD51 may be a potential marker for clinical diagnosis and treatment of Laryngocarcinoma. It may be significant in predicting clinical stage, pathologic grade and metastasis.

To explore the significance of PML expression in carcinogenesis of laryngocarcinoma and nasopharyngeal carcinoma.The PML expression in 56 laryngocarcinoma tissues and 34 nasopharyngeal carcinoma tissues were examined by immunohistochemistry. Moreover, the relationship between the PML expression, with the pathologic grade,T stage and lymph node metastasis was analyzed.The PML expression decreased with the ascending of pathological tumor grade (P0.05) and with the increasing of T grade of tumor (P0.05). The expression of PML in nasopharyngeal carcinoma was lower than that in carcinoma of larynx (P0.05). However, either in laryngocarcinoma or in nasopharyngeal carcinoma, there was no statistical difference on PML expression between with and without lymph node metastasis in laryngo carcinoma tissues and nasopharyngeal carcinoma tissues.The close relationship between PML expression and the malignant parameters such as pathological grade and T grade of laryngocarcinoma and nasopharyngeal carcinoma, indicated that PML may play a role in cancer repression but not in cancer metastasis.

To study the expression of Eotaxin and Eotaxin-2 in nasal polyposis and nasal polyp and explore the different mechanism between polyposis and polyp.Fifteen cases of nasal polyposis, thirteen cases of polyp and eight cases of normal middle turbinate were studied with immunohistochemical SP method to detect the expression of Eotaxin and Eotaxin-2.There were significant differences between either two groups about the expression of Eotaxin-2 (P0.05). The expression of Eotaxin in nasal polyposis and polyp were dramatically higher than in controls (P0.05), the expression of Eotaxin between nasal polyposis and polyp was no remarkable difference (P0.05).Both of Eotaxin and Eotaxin-2 likely play key roles in the inflammatory reaction process of nasal polyposis and polyp. The different expression of Eotaxin-2 between nasal polyposis and polyp demonstrates it may be one of the main causes in the different mechanism of the two diseases.

To investigate the expressions of p53 and PCNA by inhibition of hTERT with siRNA in xenografted laryngeal squamous carcinoma in nude mice.pshRNA involved fluorescein gene was synthesized according to the sequence of hTERT cDNA. The xenografted tumor in nude mice was established by using human laryngeal squamous carcinoma cell line (Hep-2). pshRNA packaged by METAFECTENE was transfected into the transplantation tumors. The volumes of the subcutaneous tumor were determined. Fluorescence expression in the tumors was detected by confocal microscopy. The expressions of p53 and PCNA were detected using immunohistochemistry.The subcutaneous tumors appeared in all the nude mice after transplantation five days later. The diameter of subcutaneous tumors become 5-7 mm after transplantation. Many green fluorescent cells were observed by confocal microscopy after transfection of pshRNA and blank plasmid. The growth of laryngeal squamous carcinoma in nude mice was significantly decreased as compared with the control group. The proliferation activity was inhibited by 76.50%. The expressions of p53 was upregulated and the expression of PCNA was downregulated.The transfection of pshRNA could significantly induce inhibitory effects on growth of laryngeal squamous carcinoma in nude mice and the expression of PCNA could be effectively downregulated. The inhibition of telomerase reverse transcriptase (hTERT) mRNA could upregulate the expression of p53 and promote the apoptosis of carcinoma cells as well as inhibit the proliferation of laryngeal squamous carcinoma cells.

To investigate nuclear factor-kappaB/p65(NF-kappaB/p65) and cyclooxygenase-2 (COX-2) gene expressions and clinical significance in human laryngeal squamous cell carcinoma, at the same time to investigate the relationship between the two gene expressions.Forty-one samples of laryngeal squamous cell carcinoma and ten samples of vocal cord polyps were assayed using immunohistochemical SP method for detection of NF-kappaB/p65 and COX-2 protein expression level.There is significant difference between the laryngeal carcinoma group and vocal cord polyps group about NF-kappaB/p65 and COX-2 protein expression level. Qualitative analysis found theres significant relationship between COX-2 and histological classification. There's significant correlation relationship between NF-kappaB/p65 and COX-2 by using spearman correlation analysis.(1)The expressions of NF-kappaB/p65 and COX-2 both raise in laryngeal carcinoma. (2) NF-kappaB/p65 perhaps promotes the expression of COX-2 in laryngeal carcinoma. (3) NF-kappaB was hoped to become the new treated target of laryngeal carcinoma.

To observe the distribution of gama-aminobutyric acid (GABA) ergic neurons in rats cochlear nuclei (CN) in pre-and post-operation of unilateral cochlea ablation.The technique of direct anti-GABA immunocytochemistry (SP) was used in this study.The GABAergic neurons existed extensively in cochlear nuclei(CN). The number was more, the volume was larger of GABAergic cells in CN and those cells were stained heavily before ablation. The number of GABAergic cells in lesion side was far less than that in intact side after unilateral cochlea ablation one to two weeks (P0.01), then the number of GABAergic cells increased after three weeks which was still less (P0.05) and almost reached normal after a month (P0.05).The changes of GABAergic neurons reflect the balance between excitatory and inhibitory of neurons. It suggests that changes in the extent of GABAergic inhibition have been shown to contribute to the reorganization of primary auditory central system after unilateral cochlea ablation in rats. The decrease and increase of the number of GABAergic-neurons are essential for this reorganization.

In order to observe the DPOAE of guinea pigs under awake and anesthetic conditions, we invesitigated the amplitude of distortion product emission and I/O function curves and made a comparative analysis of their changes under different conditions. It was found that the amplitude of DPOAE and I/O curves did not show difference in the guinea pigs under awake and anesthetics conditions. We deem it credible and simple to measure DPOAE in the guinea pigs under awake condition, which is similar to measuring DPOAE in the guinea pigs under physiological condition.

To observe the effects of systemic steroids on the infiltration of eosinophils and the expression of TGF beta 1 in nasal polyps.The SP immunohistochemical methods were applied to explore the expression of TGF beta 1 in nasal polyps before and after corticosteroids use; the optical density of positive cells in mucosal epithelia, eosinophils, glandular epithelium, vascular endothelial, matrix and mucous membrane was measured by using HPIAS-1,000 image-conduct system.The expression of TGF beta 1 was positive in nasal polyps. The number of eosinophils and the expression of TGF beta 1 was significantly reduced in nasal polyps after treated with corticosteroids compared with pre-treated.The effects of systemical steroid use on the nasal polyps may depend on decreasing the infiltration of eosinophils and the expression of TGF beta 1.