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Background: Macrophage migration inhibitory factor (MIF), originally reported as an inflammation regulating molecule, is elevated in various cancer cells, which may promote carcinogenesis. Meanwhile, ISO-1 is a potent small molecular inhibitor of MIF, which has not been investigated in nasopharyngeal carcinoma (NPC), hence the impact of ISO-1 on NPC cells remains to be illustrated. Objective: This study intended to explore the biological function of ISO-1 in NPC cells in vitro and prove a possibility of ISO-1 being a novel agent in NPC treatments. Methods: Gene expression of MIF in Head and Neck squamous cell carcinoma was obtained from The Cancer Genome Atlas (TCGA) database. Nasal pharyngeal tissues were collected from adult patients undergoing nasopharyngeal biopsy for MIF level detection. Proliferation of NPC cell lines 5-8B and 6-10B was studied using Cell Counting Kit-8 (CCK-8) assay and plate-colony-formation assay, apoptosis was determined by flow cytometry and TUNEL staining, migration and invasion capacities were measured by wound-healing assay and transwell assay, all to explore the function of ISO-1 in NPC cells in vitro. Epithelial-to-mesenchymal transition (EMT) level of NPC cells was determined by Western blot analysis and immunofluorescence assay. Results: Transcript level of MIF was significantly higher in head and neck squamous cell carcinoma. Protein MIF was overexpressed in human NPC tissues compared to non-cancerous ones, and its expression could be compromised by ISO-1 in vitro. 100μM ISO-1 significantly hindered NPC cells' migration and invasion capacities in vitro but acted relatively poorly on proliferation and apoptosis. Immunofluorescence assay and Western blotting implied a downregulated EMT level through TGF-β/Smad4 axis in ISO-1 treated NPC cells compared to the vehicle Conclusion: This study indicated that MIF antagonist ISO-1 holds an impact on NPC progression by influencing the migration and invasion of NPC cells ISO-1 inhibits the EMT process of NPC cells through TGF-β/Smad4 axis, supporting that prudent application of ISO-1 may be a potential adjuvant treatment for NPC.

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.

Head and neck squamous cell carcinoma (HNSCC) is the sixth most common cancer in the world, yet current treatment options are associated with limited success. The aim of the present study was to investigate the expression of ferritin in HNSCC and clarify whether it may serve as a biomarker for predicting HNSCC metastasis. The chemiluminescent immunoassay method was used to investigate the differences in the serum ferritin (SF) levels between patients with and without tumors, and between HNSCC with and without lymph node metastasis. The iron content and expression levels of ferritin were detected to verify the differences between tumor and normal tissues, and between HNSCC without and with lymph node metastasis. Data from the Gene Expression Omnibus (GEO) dataset was used to support the aforementioned results. No statistically significant difference in the SF level was observed between patients with and without tumors. Iron content and expression levels of ferritin heavy chain (FTH) and ferritin light chain (FTL) were higher in tumor tissues compared with normal tissues. The iron content and expression levels of SF, FTH and FTL were increased in HNSCC with metastasis compared with HNSCC without metastasis. The GEO dataset further verified the results and reported that the expression level of FTH was correlated with the prognosis of patients with HNSCC. Ferritin may not be a biomarker for the early diagnosis of HNSCC. However, an association exists between the expression level of ferritin and HNSCC cervical metastasis. SF may be a potential biomarker for predicting cervical lymph node metastasis in patients with HNSCC.

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.

IUR is a recently identified oncogenic lncRNA in leukaemia, while its roles in nasopharyngeal carcinoma (NPC) are unclear. We aimed to explore the possible involvement of IUR in NPC. IUR and PTEN were downregulated, while miR-144 was upregulated in NPC. In addition, IUR was inversely correlated with miR-144 and positively correlated with PTEN. In NPC cells, overexpression of IUR resulted in a downregulated expression of miR-144 and an upregulated expression of PTEN. Overexpression of miR-144 led to a downregulated expression of PTEN and attenuated the effects of overexpression of IUR. Cell proliferation assay showed that overexpression of IUR and PTEN resulted in decreased NPC cell proliferation rate. Overexpression of miR-144 played an opposite role and attenuated the effects of overexpression of IUR. In conclusion, IUR can downregulate miR-144 to upregulate PTEN in NPC, therefore inhibiting NPC cell proliferation.

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.

Summary Allergic rhinitis is thought to be an allergic disease associated with immunoglobulin (Ig)E-mediated immune response, characterized by increased T helper type 2 (Th2) cytokine production, elevated eosinophil levels in the nasal mucosa and induced nasal secretions. MicroRNA (miRNA) microarray data revealed that the expression level of miR-466a-3p was significantly decreased. Notably, GATA binding protein (GATA-3) was identified as one of its target genes through miRNA target prediction web tools. The expression levels of miR-466a-3p were altered by mimics and lentivirus both in vivo and in vitro, similar to those of GATA-3. Furthermore, the symptoms and histology of allergic rhinitis as well as the levels of serum IgE and interleukin (IL)-4 were examined in different groups of mice. Interestingly, the results for lentiviral miR-466a-3p-treated allergic rhinitis mice were relatively similar to normal mice, compared to allergic rhinitis mice without treatment. Also, miR-466a-3p negatively regulated GATA-3 expression in allergic rhinitis mice, indicating the participant of Th2-cell responses in allergic rhinitis. Taken together, our findings highlight a new perspective on the role of miR-466a-3p in allergic rhinitis. In addition, this study provides a theoretical framework and experimental reference for future research targeting microRNAs as therapeutic targets and diagnostic biomarkers of allergic rhinitis.

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 dietary compound phenethyl isothiocyanate (PEITC), an important tumoricidal component found in cruciferous vegetables, exhibits strong anticancer and chemopreventive effects in a variety of tumors. However, its role in human laryngeal cancer is unclear. The aim of the present study was to investigate whether PEITC exhibits anticancer properties in human laryngeal carcinoma Hep-2 cells in vitro and to identify the potential molecular mechanisms. The results showed that treatment of Hep-2 cells with PEITC significantly inhibited cell proliferation in a dose- and time-dependent manner, promoted apoptosis with concurrent G2/M cell cycle arrest and inhibited cell invasion in a dose-dependent manner. These effects were accompanied by significant alterations in the expression levels of key proteins associated with pro-survival signaling pathways, including PI3K, Akt, ERK, NF-κB, Bcl, Bax, cyclin B, CDK4 and CDK6. Importantly, these effects were not reflected in 16HBE normal human bronchial epithelial cells, suggesting a safe range of treatment concentrations between 0 and 10 µM PEITC. In summary, PEITC exhibited significant anticancer effects against human laryngeal cancer cells in vitro with low toxicological impact on normal bronchial epithelial cells. This was achieved through dysregulation of key proteins involved in the occurrence and development of tumors, thereby offering a valuable contribution to future strategies for the treatment and screening of patients with laryngocarcinoma.

The auditory brainstem response (ABR) wave I threshold, latency and amplitude are insensitive to spiral ganglion neurons (SGNs) degeneration, but are sensitive to the degeneration of Schwann cells and can estimate the status of Schwann cells in a neural degeneration mouse model. The thorough pre-operative ABR assessment would be helpful in predicting cochlear implant performance.This study aimed in finding a non-invasive electrophysiological method to evaluate the status of the auditory nerve and the Schwann cells in sensorineural hearing loss (SNHL) and auditory neuropathy (AN) ears, and providing useful information for candidates screening and outcome prediction in cochlear implantation.The frequency-specific acoustic ABR was recorded in mice. The immunohistochemical staining was performed to detect the SGNs and Schwann cells in mice cochlea. The correlations between ABR wave I metrics and SGNs, Schwann cells were investigated.In SNHL and AN mice cochlea, statistically significant correlations between ABR wave I thresholds, latencies and amplitudes at 8, 16, and 32 kHz and their corresponding SGNs densities were found only in wave I amplitude at 8 kHz. While the ABR wave I metrics at all three frequencies showed strong significant correlations with their corresponding Schwann cells densities.

Allergic rhinitis (AR) is primarily caused by a T helper cell (Th)1/Th2 imbalance. In a murine AR model of a previous study, the serum ovalbumin (OVA)-sIgE concentration was high, whereas microRNA (miR)-135a was lowly expressed in the nasal mucosa. The abnormal expression pattern of miR-135a coincided with highly expressed endogenous factors, including GATA binding protein (GATA)-3 and interleukin (IL)-4, and lowly expressed factors, including T-box expressed in T cells (T-bet) and interferon (IFN)-γ. We hypothesized that miR-135a may play an important role in immune regulation in AR mice. In the present study, AR was induced by OVA in the mice. Two groups of the AR mice were treated with a miR-135a mimic and a mimic control, respectively. The serum and nasal mucosa were collected for analysis. Following miR-135a application, the serum OVA-sIgE concentration was significantly reduced. In the nasal mucosa, the expression levels of miR-135a were higher, the mRNA and protein expression levels of GATA-3 and IL-4 were lower, and the expression levels of T-bet and IFN-γ were higher. The miR-135a corrected the Th1/Th2 imbalance in the AR mice. Findings of this study may provide a basis for novel genetic treatments in addressing allergic diseases.

Lactoferrin (LF) can downregulate allergic airway inflammation in asthma. However, the in vivo effect of exogenous LF on allergic rhinitis (AR), a disease attributed to airway inflammation, has yet to be determined. We investigated the effect of intranasal administration recombinant human (rh) LF and its underlying mechanisms on AR in BALB/c mice. Multiple parameters of allergic responses were evaluated to determine the effect of rhLF. We found that the number of eosinophils and goblet cells, as well as mRNA and protein expression of type 2 helper T (Th2), Th17 and regulatory T (Treg) cells in the nasal cavity, was significantly upregulated in AR mice compared with the controls, Conversely, administration of rhLF prior to or after intranasal ovalbumin challenge markedly downregulated these same parameters. Th1-specific mRNA and protein expression in the nasal cavity of the controls was not different from that in AR mice, but expression significantly increased with rhLF treatment. The mRNA and protein expression of endogenous LF in the nasal cavity was significantly downregulated in AR mice compared with the controls. However, after rhLF treatment, endogenous LF mRNA and protein expression was significantly upregulated. Exogenous rhLF inhibited allergic inflammation in AR mice, most likely by promoting the endogenous LF expression and skewing T cells to a Th1, but not a Th2 and Th17 phenotype in the nasal mucosa. Our findings suggest that rhLF treatment may be a novel therapeutic approach for prevention and treatment AR.

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.

TERT is the main functional unit of telomerase, which maintains telomere length and chromosome structure stability. TERT has been shown to act as a key factor in various biological processes, such as cell proliferation, via uncharacterized mechanisms. We transfected HEp-2 laryngeal carcinoma cells with a TERT overexpressing adenovirus (Ad-TERT) and TERT shRNA silencing adenovirus (Ad-sh-TERT), and examined the effect on TERT and the AP-1 transcription factor subunits c-Fos and c-Jun using RT-PCR and western blot analysis. TERT mRNA expression was quantified using RT-PCR in 24 human laryngeal carcinoma samples, and TERT protein co-expression with AP-1 was investigated in a human laryngeal carcinoma tissue microarray using quantum-dot based immunofluorescence. The effect of specific ERK and p38 inhibitors on ERK, p38, c-Jun and c-Fos phosphorylation was investigated in TERT-overexpressing HEp-2 cells. TERT overexpression led to increased TERT, c-Jun and c-Fos mRNA and protein expression and increased cell proliferation, while TERT silencing had the opposite effects. TERT mRNA expression levels were positively correlated with c-Fos and c-Jun mRNA in human laryngeal carcinoma tissue. TERT and AP-1 protein were expressed at high levels and positively correlated in laryngeal carcinoma tissues. Treatment of TERT-overexpressing HEp-2 cells with specific p38 and ERK inhibitors indicated that TERT modulates the expression and phosphorylation of the AP-1 subunits c-Jun and c-Fos through the p38 and ERK signaling pathways. In conclusion, the results of this study indicate that TERT is capable of promoting cell proliferation via activation of the AP-1 subunits, c-Jun and c-Fos, in laryngeal carcinoma cells.

A mouse model of allergic rhinitis (AR) was prepared, and exogenous surfactant protein A (SP-A) was given by an intranasal route to study its mechanism and effects in the mice.Sixty male BALB/c mice were randomly divided into a normal control group, a group with AR (AR group), and a group with AR that was given SP-A (treatment group).A mouse model of AR was successfully established. Enzyme-linked immunoassay showed that the level of ovalbumin-specific immunoglobulin E in the AR group was significantly higher than those in the treatment and control groups (p0.05), whereas the levels were not significantly different (p0.05) between the treatment and control groups. Hematoxylin-eosin staining showed typical allergic injury of the nasal epithelium in the AR group, and the number of eosinophils that migrated into the nasal tissue in the AR group was significantly greater than those measured in the treatment and control groups (p0.05). Western blotting and real-time quantitative polymerase chain reaction testing revealed that the type 2 helper (Th2) cytokine (interleukin 4 and interleukin 5) levels were highest in the AR group, followed by the treatment and control groups, with significant differences between each of the groups (p0.05). Significant differences were found in the levels of nasal mucosa type 1 helper (Th1) cytokines (interferon gamma, interleukin 12) among the AR, treatment, and control groups; the highest levels were found in the control group, and the lowest levels were detected in the AR group (p0.05).Exogenous SP-A had a significant therapeutic effect in mice with AR, and its mechanisms of action included inhibition of the differentiation of Th2 cells in the nasal mucosa, reduced levels of Th2 cytokines, and increased levels of Th1 cytokines. Together, these effects corrected the Th1/Th2 imbalance, inhibited the increase of specific immunoglobulin E production, effectively reduced the symptoms of AR, and inhibited the development of AR.

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.

Purpose To explore the influences of telomerase and alternative lengthening of telomeres mechanism on telomere length and laryngeal squamous cell carcinoma in vitro and in vivo. Materials and methods Short hairpin RNA expression vectors targeting the messenger TERT, TRF2, RAD51 and NBS1 were constructed. The mRNA and protein expression of targeted genes in human laryngeal squamous carcinoma cell line HEp-2 was evaluated by reverse transcription polymerase chain reaction and Western blotting separately. The length of telomere was analyzed by fluorescent in-situ hybridization. Cell viability was examined by cell counting Kit-8. Effects on tumor growth were also investigated in vivo. Results The transfection of multiple short hairpin RNAs expression plasmid significantly inhibited the mRNA and protein expression of related genes. Silence of alternative lengthening of telomeres mechanism and telomerase mechanism related genes resulted in the shortening of telomere length in HEp-2 cell. However, silence of alternative lengthening of telomeres mechanism related genes could shorten the telomere length but had no significant difference. Both simultaneously and separately blocking telomerase mechanism and alternative lengthening of telomeres mechanism resulted in reduction of tumor cell viability. Silence of alternative lengthening of telomeres mechanism and telomerase mechanism related genes inhibited the tumor growth in vivo. Conclusions The inhibition of telomere related gene may be a promising strategy for the treatment of laryngeal squamous cell carcinoma.

Calpains are a family of intracellular cysteine proteases involved in various biological processes. Previously, the family was identified to have abnormal expression in several types of malignant tumor. Calpain 6 was less well known; however, it was recently identified to be involved in the carcinogenesis of certain types of malignant tumor. However, the expression of calpain 6 in head and neck squamous cell carcinoma (HNSCC) remains unclear. A total of six datasets from the Gene Expression Omnibus (GEO) was analyzed and an association between calpain 6 expression levels and HNSCC was identified, with the expression of calpain 6 observed to be significantly decreased in HNSCC (P

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.

The Notch signaling pathway plays an important role in the proliferation and differentiation of cells. Although recent studies have shown that Notch plays a role in the mechanisms of cisplatin resistance, the mechanism by which Notch plays roles in intrinsic or acquired cisplatin resistance remains unclear. In the present study, poorly differentiated nasopharyngeal carcinoma cells were treated with a γ-secretase inhibitor (DAPT), which led to a decrease in the Notch intracellular domain and inhibition of Notch signaling. Treatment was not sufficient to induce pronounced apoptosis of CNE-2 cells, but did result in the down-regulation of the P-glycoprotein and ERCC1 protein. In contrast, the combined treatment of DAPT and cisplatin induced substantial cell apoptosis compared to cisplatin treatment alone.

RNA interference (RNAi) is an evolutionary conserved mechanism for specific gene silencing. There are currently numerous cancer therapy clinical trials based on RNAi technology. Using an adenoviral system as a delivery mediator of RNAi, we investigated the therapeutic effects of targeting three genes simultaneously in vitro and in vivo. In this study, we constructed an recombinant adenoviral shRNA expression system as Adv-pEGFP-shVEGF-shTERT-shBcl-xl for multi-genes silencing. Our results showed that the adenoviral vector can achieve above 90% of transfection efficiency and induced obvious apoptosis in CNE-2 cell both in vitro and in vivo compared with targeting the TERT alone or controlled group.

The development of allergic rhinitis is considered to be determined by the interaction between genetic and environmental factors. Surfactant protein A (SP-A), a member of the collectin family of proteins, plays an important role in immune regulation. The purpose of this study was to investigate the association between SP-A polymorphisms and allergic rhinitis. We conducted a case–control association study on a Chinese Han population, comprising 216 adult individuals with AR and 84 healthy controls. A total of 9 single-nucleotide polymorphisms (SNPs) mapped to the SP-A were genotyped using PCR-based molecular identification methods. The frequency of A allele at amino acid 223 in the patient group was significantly higher than that in the control group after correcting for multiple testing (P = 0.006). The 1A2 allele haplotype in SFTPA2 was associated with decreased risk for allergic rhinitis, after applying Bonferroni corrections (P = 0.003). However, genetic variants of the SFTPA1 genes were not found to be associated with AR. In addition, no significant associations were established between any of the 9 SFTPA gene polymorphisms and the skin-prick test responses (P > 0.05). Further, no association was established between the 9 SNP loci and the levels of total serum immunoglobulin E (IgE) (P > 0.05). These results indicate that the gene polymorphism at the residue 223 in the carbohydrate recognition domain of SFTPA2 may be a genetic marker for the development of AR in the adult Chinese Han population.

Background and purpose:Telomere length must be maintained throughout cancer cell progression and proliferation. In most tumours, telomerase activity maintains telomere length. Therefore, telomerase is a target for cancer treatments. However, some cancer cells maintain telomere length through an alternative mechanism termed ‘alternative lengthening of telomeres’. To determine how telomerase inhibition relates to the initiation of the alternative lengthening of telomeres pathway, we investigated telomerase activity and telomere maintenance in Hep-2 cells with and without reduced telomerase activity.Materials and methods:We investigated telomerase activity levels in a normal Hep-2 cell line and in residual cells following telomerase inhibition treatment. Additionally, we looked for expression of a marker protein for the alternative lengthening of telomeres mechanism.Results and conclusions:In the residual cells, telomerase activity was eliminated. However, these cells had higher levels of the alternative lengthening of telomeres biomarker, suggesting an alternative mechanism for telomere maintenance following telomerase inhibition. These results could have a major impact on the design of new cancer treatments.

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.

Telomerase activity is mainly regulated by the human telomerase reverse transcriptase (hTERT) gene. Our objective was to investigate the effect of short hairpin RNA (shRNA) directed against hTERT mRNA on telomerase activity in laryngeal cancer cells (Hep-2), nasopharyngeal carcinoma cells (NEC), and human bone marrow mesenchyme stem cells (hMSCs). Short hairpin RNA expression vectors targeting the messenger RNA of hTERT were constructed. Cells were treated with shRNA expression vectors directed against hTERT mRNA and control vectors that included mismatched shRNA. We found that treatment of special shRNA expression vectors induced significantly decrease in hTERT expression, telomerase activity, and cell viability in Hep-2 and NEC cells. In contrast, the shRNA control showed none of these effects. And none of these effects appeared in hMSCs cells. Our results suggest that shRNA against hTERT mRNA inhibits telomerase activity and cell viability through suppression of the hTERT expression in cancer cells. And this treatment has no side effect on healthy cells lack of telomerase activity. RNA interfering technology may be a promising strategy for the treatment of cancers.

Objective: Analysis of the correlation between telomerase and the expression of its related proteins may provide insight into the molecular mechanism of nasopharyngeal carcinogenesis. We investigated the effect of short hair pin ribonucleic acid (RNA) specific for human telomerase reverse transcriptase messenger RNA on the expression of the proteins c-myc (the transcription factor c-myc is a shortlived nuclear phospho-protein involved in cell proliferation and differentiation, belongs to the myc family), proliferating cell nuclear antigen and Caspase-3 in nasopharyngeal carcinoma cells.Methods: Short hairpin RNA expression vectors targeting the messenger RNA of human telomerase reverse transcriptase were constructed. Cells were treated with the short hairpin RNA expression vectors targeting human telomerase reverse transcriptase or vectors that included mismatched short hairpin RNA, and telomerase activity was measured by telomeric repeat amplification enzyme-linked immunosorbent assay. Cell viability was examined using the 3-(4,5-dimethyl thizol-2-yl) 2,5-diphenyl tetrazolium bromide assay. The expression of the three proteins (c-myc, proliferating cell nuclear antigen and Caspase-3) was determined by Western blotting.Results: Short hairpin RNA specific for human telomerase reverse transcriptase messenger RNA significantly inhibited telomerase activity. In addition, the expression of and proliferating cell nuclear antigen were both inhibited, while the expression of Caspase-3 was up-regulated.Conclusions: Our results suggest that short hairpin RNA directed against human telomerase reverse transcriptase inhibits cell viability by regulating telomerase activity and its related proteins expression in nasopharyngeal carcinoma cells. Therefore, RNA interference technology may be a promising strategy for the treatment of nasopharyngeal cancer.

A number of different approaches have been developed to inhibit telomerase activity in human cancer cells. In this study, the effect of antisense oligonucleotides (ODNs) by targeting human telomerase reverse transcriptase (hTERT) mRNA in a laryngeal cancer cell line (Hep-2) was investigated. A 20mer antisense oligodeoxynucleotide targeting the most open part of hTERT mRNA (anti-hTERT) and a mismatched control sequence were synthesized. Cells were treated daily with oligonucleotides for up to 72 hours. hTERT mRNA expression was measured by the reverse transcription polymerase chain reaction (RT-PCR) assay; telomerase activity by the telomerase PCR ELISA assay kit (TRAP; Boehringer Mannheim, GmbH, Mannheim, Germany). Cell viability after administration of ODNs was determined using the MTT assay. Morphological changes were examined by haematoxylin and eosin staining. The cell cycle was analyzed using flow cytometry. It was found that antisense treatment induced a decrease in hTERT mRNA expression, telomerase activity, cell growth rate, cell viability, and an increase in apoptosis. The results suggest that inhibition of telomerase activity in Hep-2 cells by short-term antisense treatment against the mRNA of hTERT results in apoptotic cell death. The treatment with anti-hTERT may be useful as a treatment modality for laryngeal squamous carcinoma.

Nasopharyngeal carcinoma is a common malignant tumor in the head and neck. Because of frequent recurrence and distant metastasis which are the main causes of death, better treatment is needed. Indole-3-carbinol (I3C), a natural phytochemical found in the vegetables of the cruciferous family, shows anticancer effect through various signal pathways. I3C induces G1 arrest in NPC cell line with downregulation of cell cycle-related proteins, such as CDK4, CDK6, cyclin D1 and pRb. In vivo, nude mice receiving I3C protectively or therapeutically exhibited smaller tumors than control group after they were inoculated with nasopharyngeal carcinoma cells. The expression of CDK4, CDK6, cyclin D1 and pRb in preventive treatment group and drug treatment group both decreased compared with the control group. We conclude that I3C can inhibit the growth of NPC in vitro and in vivo by suppressing the expression of CDK and cyclin families. The drug was safe and had no toxic effects on normal tissues and organs.

To establish the rat model of depression and observe its morphological changes and nerve growth factor (NGF) expression in nasal mucosal.Thirty SD rats were divided into depression model group and normal controls with 15 rats in each group. The rat depression model were made by application of chronic unforeseeable medium stress stimulating. Behavior change were observed with open box experiment(open field) and humoral consumption-experiments (sucrose consumption), After being sacrificed, the nasal mucosal were taken for HE and NGF immunohistochemical study. The difference of nasal mucosal NGF protein between two groups were examined with medical digital image analysis technology.There were no swelling and inflammatory cells infiltrating in nasal mucosal of normal group, but 53% nasal mucosal of depression model were observed such morphological changes. Th NGF immunohistochemical staining were negative in normal group but positive in 53% depression model group. There were significant differences between two groups (P0.01).Depression rats may appear rhinitis symptoms under long-term stress stimulating. NGF may exert immunoregulatory effects, stimulate the immune cell proliferation and gathering, induce various media release, increase the sensitivity of nerve fibers itself and the producing of neuropeptide, promote the formation of nerve source inflammation and increase the reactivity of nasal mucosa.

To evaluate the effects of radiotherapy and surgery on carcinoma of the middle ear.A review of five published literatures was conducted according to defined selection criteria by the Review Manager 5.0 statistical software.There were no systematic reviews or large-scale RCTs between radiotherapy and symptomatic treatment containing surgery and radiotherapy for carcinoma of the middle ear.Radiotherapy and symptomatic treatment for carcinoma of the middle ear have no obvious differences. The radiotherapy is the first choice for the treatment of squamous cell carcinoma of the middle ear.

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.

In most human malignancies, telomere homeostasis is maintained by the reactivation of telomerase. While inhibiting telomerase provides a novel approach to the treatment of many cancers, telomere maintenance can occur in the absence of telomerase activity by the alternative lengthening of telomeres (ALT) mechanism. Therefore, it must be determined if inhibiting telomerase selects for cancer cells that activate ALT. Here, we report that Hep-2 cells that survived anti-telomerase treatments showed sustained proliferation in culture with down-regulated human telomerase reverse transcriptase (hTERT) expression and significantly enhanced levels of ALT-specific promyelocytic leukemia (PML) bodies. Analysis of the telomere lengthening kinetics also demonstrated elevated telomeric sister-chromatid exchange (T-SCE) in surviving Hep-2 cells, consistent with their long and heterogeneous telomeres. Similar to ALT cells, the surviving cells showed evidence of ALT telomere homeostasis. Furthermore, proteomic analysis identified several proteins differentially expressed between the untreated Hep-2 cells and surviving cells that may provide new insight for understanding these two telomere maintenance mechanisms. Thus, the findings in this study may help to improve telomerase-based therapy for cancer.

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.

Objective:To research the protective effect of melatonin against gentamicin ototoxicity.Methods:Guinea pigs were randomly divided into four groups. The first group received intramuscular gentamicin (120 mg/kg body weight/day) for 17 days. Over the same time period, a second group simultaneously received intramuscular gentamicin (120 mg/kg body weight/day) plus (on the other side) intramuscular melatonin (0.3 ml kg body weight/day). Two groups of controls were treated for 17 days with either intramuscular melatonin or intramuscular saline. After the 17 days, each animal underwent distortion product otoacoustic emission testing (both ears). The guinea pigs were sacrificed by decapitation just after the final injection. Their cochleae were used to produce a tissue section, surface preparation and scanning electron microscope preparation.Results:Distortion product otoacoustic emission testing indicated gentamicin-induced hearing loss at 3, 4, 6 and 8 kHz in gentamicin-treated animals. Animals receiving melatonin co-therapy had significantly attenuated hearing loss and their cochleae showed lower rates of outer hair cell loss (comparing the same cochlear turns), compared with gentamicin-treated animals (p Conclusion:These findings confirm the occurrence of outer hair cell loss after gentamicin treatment, and the attenuation of such loss following simultaneous melatonin injection, using the method of morphological evaluation. These results suggest that melatonin protects against gentamicin ototoxicity by interfering with cytotoxic mechanisms.

Analysis of the correlation between telomerase and expression of its related proteins may provide insight into the molecular mechanism of nasopharyngeal carcinogenesis. Here, we investigate the effect of short hairpin RNA (shRNA) specific for human telomerase reverse transcriptase (hTERT) mRNA on the expression of proliferating cell nuclear antigen (PCNA) and Caspase-3 in nasopharyngeal carcinoma (CNE) cells.shRNA expression vectors targeting the mRNA of hTERT were constructed. Cells were treated with the constructed expression vectors and telomerase activity was measured by telomeric repeat amplification ELISA (TRAP-ELISA). Cell viability was examined using the MTT assay. Cell apoptosis was detected by TUNEL method. The expression of PCNA and Caspase-3 proteins, was determined by Western blotting.shRNA specific for hTERT mRNA significantly inhibited telomerase activity, suppressed cell viability and induced apoptosis in CNE cells. In addition, the expression of PCNA was inhibited, while the expression of Caspase-3 was up-regulated.Our results suggest that shRNA directed against hTERT inhibits cell viability by regulating telomerase activity and its related protein expression in NPC cells. Therefore, RNA-interfering technology may be a promising strategy for the treatment of nasopharyngeal cancer.

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 develop an RNA-interference (RNAi) approach involving the hitting of multiple targets by a recombinant plasmid and evaluate its antitumor effect on laryngeal squamous carcinoma in vitro and in vivo.A plasmid containing 3 different short hairpin RNA (shRNA) segments termed pEGFPshVEGF-shTERT-shBcl-xl was constructed. Plasmids containing single shRNA against each target (VEGF, TERT, BCL-xl alone) individually were also constructed as control. Cells were treated with these plasmids. The expression of targeted genes as well as apoptosis of tumor cells were evaluated after treatment with multiple shRNA vectors or control vectors. The mRNA and protein expression were determined by RT-PCR and western blotting. Cell viability was examined using the MTT assay. Apoptotic morphological alterations were observed by Hoechst staining and electron microscopy. The in vivo antitumor effect was characterized in a nude mice model of laryngeal squamous carcinoma.We demonstrated that a recombinant plasmid containing multiple shRNAs could effectively and simultaneously inhibit VEGF, TERT and Bcl-xl mRNA and protein expression in the HEp-2 cells; the plasmid containing the 3 different shRNAs exhibited a potent antitumor effect on LSCC both in vitro and in vivo, and could much more effectively induced cell apoptosis than each single shRNA. We also demonstrated that the simultaneous blockage of these 3 genes have a better inhibitory effect on human HEp-2 cells than the blockage of each single shRNA.Our study demonstrates that the application of vector-based RNAi technology that involves hitting multiple targets will be a promising therapeutic modality in the gene therapy of human laryngeal cancers; furthermore, it provides experimental evidence for the clinical application of this technology in the future.

In our previous work, we constructed short hairpin RNA (shRNA) expression plasmids that targeted human telomerase reverse transcriptase (hTERT) messenger RNA, and we demonstrated that these vectors could inhibit the growth of cancer cells by 76.5% in vivo. In order to maximize the efficiency and versatility of the vector-based siRNA approach, here we have constructed multiple shRNA expression vectors that simultaneously targeted 3 different genes in cancer cells, and then investigated their effect on human laryngeal squamous carcinoma (Hep-2) in vivo.We found that expression of multiple shRNAs led to a significant reduction in VEGF, hTERT and Bcl-xl mRNA and protein expression. Tumor growth curves showed that those tumors treated with the shRNAs were obviously smaller than control, nontransfected tumors. Analysis at 14 days following the final injection showed a tumor inhibition ratio of 91.2%. However, the control shRNA vectors showed none of these effects.Short hairpin RNA expression vectors targeting the mRNA of VEGF, hTERT and Bcl-xl were constructed and subsequently transfected by direct injections into the tumors formed by Hep-2 cells implanted in nude mice. The expression of targeted genes as well as apoptosis of tumor cells were evaluated after treatment with multiple shRNA vectors or control vectors.Our results demonstrate that the application of vector-based RNAi technology that involves blocking multiple targets will be a promising therapeutic modality in the gene therapy of human cancers. The data strongly suggest that simultaneously blocking multiple genes in human cancers using an RNAi approach should be considered in cancer therapy.

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 observe the changes of GABA and GABAergic neurons in rat inferior colliculus following unilateral cochlear damage and explore the function and significance of GABA in reorganization of auditory center after deafferentation.Twenty sprague dawley rats were divided into four groups randomly. The technique of direct anti-GABA immunocytochemistry (SP) was used in this study. The quantity of GABA was measured by 835-50 type Amino Acid Automatism Analyzer and compared at one week, two weeks and one month after unilateral cochlear ablation, respectively.Compared with controlled groups, the quantity levels of GABA decreased and the number of GABAergic neurons also reduced one week after unilateral cochlear damage and there was a significant difference in GABA between two groups( P0. 05); after one month, the quantity of GABA and the number of GABAergic neurons almost reached the normal level, the differences were no significant.The dynamic change of GABA in rat inferior colliculus reflected the neuronal activity, which implied that GABA may play an important role in reorganization of auditory center after unilateral cochlear damage.

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 the effect of inhibiting human telomerase reverse transcriptase (hTERT) on expression of C-myc protein by RNA interference (RNAi) in the larynx cancer cell line, Hep-2.The primary structures of hTERT cDNA were found in GeneBank. Then the structure analyses were done according to the strategy of RNAi, which determined the specific base sequences to design shRNA plasmid. One type of plasmid, pshRNA1, involved in fluorescein gene was synthesized based on the specific base sequence. Control pshRNA2-a random sequence-were also constructed. METAFECTENE was used as the transfect ion reagent. Cells were treated daily with pshRNA1-2 or normal culture medium respectively. After administration of pshRNA1-2, hTERT mRNA was detected by RT-PCR, hTERT protein and C-myc protein were examined by Western Blot.The expression of hTERT mRNA and protein were both significantly decreased after treated by pshRNA1 (P0.05). The expression of C-myc protein was significantly increased after treated by pshRNA1 (P0.01).The inhibition of hTERT expression could increase the expression of C-myc protein in Hep-2 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.