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To investigate the role of Tregs and their subtypes in the treatment of allergic rhinitis with allergen immunotherapy (AIT) as well as the underlying mechanism.1. Thirty-one healthy controls, 29 Allergic rhinitis (AR) patients and 16 AR patients treated with AIT were recruited. The total nasal symptom scores (TNSSs) were calculated. The serum levels of IgE, IL-2, TNF-α, IFN-γ, IL-4, IL-5, IL-6, IL-10 and IL-17 were measured. 2. Changes in the proportions of CD4+ T cells, Treg cells, Treg subtypes and Th1/Th2/Th9/Th17/Tfh cells in the peripheral blood of the subjects in the three groups were measured. 3. The correlations of Treg cells, Treg subtypes and TNSS with the levels of various cytokines in the AR group and AIT group were analysed.1. Compared with the control group, the TNSS and IgE, IL-5 and IL-6 levels in the AR group were significantly increased, while the IL-2, IFN-γ and IL-10 levels were significantly decreased (P 0.05). Compared with the AR group, the TNSS and IgE, IL-5 and IL-6 levels in the AIT group were significantly decreased, while the IL-2, IFN-γ and IL-10 levels were significantly increased (P 0.05). 2. Compared with the control group, the proportions of Tregs, GATA3+ Tregs and Th1 cells in the AR group were significantly reduced, while the proportions of PU-1+ Tregs, T-bet+ Tregs and Th2 cells were significantly increased (P 0.05). Compared with the AR group, the proportions of Tregs and Th1 cells in the AIT group were significantly increased, while the proportions of PU-1+ Tregs and Th2 cells were decreased (P 0.05). 3. Correlation analysis showed that Treg cell proportions were negatively correlated with the TNSS, sIgE levels, IL-5 levels and IL-6 levels but positively correlated with the IL-2 and IL-10 levels (P 0.05). PU-1+ Treg cell proportions were positively correlated with the TNSS, sIgE levels, IL-5 levels and IL-6 levels but negatively correlated with the Treg cell proportions, IL-2 levels and IL-10 levels (P 0.05).AIT can reduce the proportions of PU-1+ Treg subtypes in AR patients. PU-1+ Treg cell numbers can potentially be used as an indicator to monitor the therapeutic effect of AIT on AR.

The aim of this study was to investigate the role and mechanism of Notch2-dependent GATA3+ Treg cells in allergic rhinitis (AR). Samples were collected from patients in the control and AR groups to detect differences in the numbers of GATA3+ Treg cells and their intracellular Notch2 levels. The effects of Notch2 on GATA3+ Treg cell differentiation and function in vitro were detected. AR mice were subjected to adoptive transfer of GATA3+ Treg cells to detect changes in the allergic inflammatory response and Th2 cells. Mice with Treg cell-specific knockout of Notch2 were constructed, and an AR model was established to detect the changes. The number of GATA3+ Treg cells and intracellular Notch2 expression in peripheral blood of the AR group were decreased compared with the controls (P 0.05), and the number of GATA3+ Treg cells was significantly negatively correlated with the level of allergen-specific IgE (sIgE; P 0.01). In vitro experiments showed that Notch2 promoted the differentiation and immunosuppressive function of GATA3+ Treg cells, and Notch2 directly promoted GATA3 transcription in Treg cells (P 0.05). Animal experiments indicated that adoptive transfer of GATA3+ Treg cells reduced the allergic inflammatory response in AR mice (P 0.05). The number of GATA3+ Treg cells was decreased in gene knockout mice (P 0.05), and autoimmune inflammation was observed. After modeling, the allergic inflammatory response was further aggravated (P 0.05). Overall, our findings indicate that Notch2 alleviates AR by specifically increasing GATA3+ Treg cell differentiation. Notch2 expressed in Treg cells is expected to be a new therapeutic target for AR.

Background and objective: Maggots are the larval stage of Lucilia sericata and have strong antibacterial activity and immunomodulatory effects. The objective of our study was to investigate whether maggot extracts can modulate regulatory T cells (Tregs) and treat allergic rhinitis (AR). Methods: Mice were randomly assigned to five groups (n=6/group): normal, AR, Maggot, AR+ Maggot, and AR+ dexamethasone (DXM). The Total Nasal Symptom Score (TNSS), ovalbumin (OVA)-sIgE titers, histopathological characteristics and Th1-/Th2-/Th17-related cytokine levels were evaluated. The expression of T-bet, GATA3, RORγt and Foxp3 in the spleen and nasal mucosa of mice was detected, and the proportion of differentiated Tregs in the spleen of mice was determined. In addition, the effects of maggot extracts on the expression level of Foxp3 and the differentiation of Tregs in vitro were studied. Histological evaluation of the potential toxicity was also performed. Results: Compared with the AR group, the AR+ Maggot group showed reduction in histopathological inflammation, downregulated OVA-sIgE titers, and restoration of the imbalance in cytokine profiles (P0.05). In vitro, maggot extracts promoted the expression of Foxp3 and differentiation of Tregs in a dose- and time-dependent manner (P

The aim of this study was to evaluate factors affecting the recurrence of positive RT-PCR results. By performing a retrospective analysis, we evaluated the clinical data of recurrent positive coronavirus disease 2019 (COVID-19) patients in multiple medical institutions in Wuhan. We recruited COVID-19 patients who were hospitalized from January 1 to March 10, 2020, in three tertiary hospitals in Wuhan, met the discharge criteria and received at least one additional nucleic acid test before leaving the hospital. According to the RT-PCR results, patients were split into a recurrent positive group (RPos group) and a nonrecurrent positive group (non-RPos group). Clinical characteristics, therapeutic schedules and antibody titers were compared between the two groups. AI-assisted chest high-resolution computed tomography (HRCT) technology was applied to investigate pulmonary inflammatory exudation and compare the extent of lung areas with different densities. This study involved 122 COVID-19 patients. There were no significant differences in age, sex, preexisting diseases, clinical symptoms, clinical classification, course of disease, therapeutic schedules or serum-specific antibodies between the two groups. A higher proportion of patients who showed pulmonary inflammatory exudation on HRCT scans were recurrent positive at the time of discharge than other patients (81.6% vs 13.7%, P < 0.01). In addition, the degree of pulmonary fibrosis was higher in the RPos group than in the non-RPos group (P < 0.05). Subpleural exudation at the peripheral edge of the lung and extensive pulmonary fibrosis at the time of discharge represent risk factors for the recurrence of COVID-19.

The practices of head and neck surgical oncologists must evolve to meet the unprecedented needs placed on our health care system by the Coronavirus disease 2019 (COVID‐19) pandemic. Guidelines are emerging to help guide the provision of head and neck cancer care, though in practice, it can be challenging to operationalize such recommendations. Head and neck surgeons at Wuhan University faced significant challenges in providing care for their patients. Similar challenges were faced by the University of Toronto during the severe acute respiratory syndrome (SARS) pandemic in 2003. Herein, we outline our combined experience and key practical considerations for maintaining an oncology service in the midst of a pandemic.

Background. Deciphering the immune characteristics within tumors and identifying the immune signals related to the prognostic factor are helpful for the treatment and management of tumor patients. However, systematic analysis of immune signatures in head and neck squamous cell carcinoma (HNSCC) remains largely unstudied. Methods. A total of 718 immune-related genes were extracted from RNA sequencing data from 519 HNSCC patients in the TCGA database, and survival analysis with integrated bioinformatics analyses was performed to build the final predictive prognosis model. Results. The 178 survival-associated genes ( P < 0.05 ) participated in important immune functions, including immune cell activation and migration. Multivariate regression analysis using 93 genes ( P < 0.01 ), together with survival-associated clinicopathological parameters, identified 35 independent prognostic factors. The most significant 8 independent factors were CD3E, CD40LG, TNFRSF4, CD3G, CD5, ITGA2B, ABCB1, and TNFRSF13b. The final prognostic model achieved outstanding predictive efficiency with the highest AUC of 0.963. Conclusion. Our prognostic model based on the immune signature could effectively predict the prognosis of HNSCC patients, providing novel predictive biomarkers and potential therapeutic targets for HNSCC patients.

Purpose The purpose of this study was to investigate the clinical features of eosinophilic hyperplastic lymphogranuloma (EHLG) in the head and neck. Materials and methods Collecting the patients who diagnose with EHLG by pathological examination. The EHLG patients with the masses involved regions, such as involved inguinal region, chest wall, abdominal wall, anterior superior iliac spine or clavicle, instead of head and neck were excluding. All of the participants will sign the informed consent form. The history data includes: clinical history, blood routine test, pathological examination, and recurrence will be collected. Results A total of 41 patients of EHLG were included. These patients predominantly presented as an enlarging and painless single or multiple masses with a history of repeated swelling. There were the complaint of itchy skin and pigmentation. The routine blood test showed that the percentage value of eosinophil increased in almost patients including 26 cases had raised absolute eosinophil count. The serum level of lgE was increased in 29 cases remarkably. With the methods of treatments, 36 patients received surgical excision, 3 patients accepted hormonotherapy, and another 2 patients for radiotherapy. The recurrence of EHLG was in 9 patients. Conclusions EHLG is a rare disease. The clinical manifestation (itchy skin and pigmentation) and increased eosinophil play critical values to the diagnosis of EHLG. Confirmed diagnosis always depends on pathological examination. Surgery is a preferred treatment, while low dose of radiotherapy is necessary for preventing relapse after operation and hormonotherapy.

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.

MicroRNAs (miRNAs) play critical roles in carcinogenesis and development of cancers. In this study, we analyzed the eccentrically expressed miRNAs in head and neck squamous cell carcinoma (HNSCC) tissues based on the miRNA-Seq data of HNSCC patients available in the Cancer Genome Atlas database. Aberrant expression of 2589 miRNAs was detected in HNSCC tissues (1128 downregulated and 1461 upregulated). The differential expression levels of the miRNAs were further validated by analysis of 25 HNSCC samples and paired control tissues and compared with the Gene Expression Omnibus database to determine the candidate miRNAs. Quantitative reverse transcription polymerase chain reaction was used to compare the expression of these candidate miRNAs between 22 fresh HNSCC tissue samples and 11 control samples. In addition, the relationship between the expression of these candidate miRNAs and Tumor, Node, Metastases staging of HNSCC was analyzed. Compared with the expression in control tissues, the levels of hsa-miR-410-3p, hsa-miR-411-5p, hsa-miR-125b-2-3p, and hsa-miR-99a-3p were significantly lower in HNSCC. According to the Cancer Genome Atlas dataset analyzed, all 4 miRNAs were shown to inhibit tumor progression (T stage), positive lymph node metastasis (N stage), and distant metastasis (M stage) in HNSCC. Kyoto Encyclopedia of Genes and Genomes analysis showed that genes regulated by these 4 miRNAs were enriched in certain pathways, including the transforming growth factor-β signaling pathway and the Hippo pathway. Enriched gene ontology terms mainly included regulation of transcription, cell proliferation, and apoptosis, which are well-characterized functions of miRNAs. Moreover, all 4 miRNAs inhibited the progression of primary tumors (T stage) and metastasis of regional lymph nodes (N stage). The top 4 aberrantly expressed miRNAs identified in this study have great clinical value in developing strategies for early diagnosis and treatment of HNSCC. More intensive studies are required to elucidate the mechanism underlying the roles of these miRNAs in HNSCC.

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 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.

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.

Mast cell (MC) degranulation is the foundation of the acute phase of allergic rhinitis (AR). Previously, downregulation of GATA binding protein 3 (GATA-3) was shown to suppress MC activation in an AR mouse model. Binding of microRNA-135a (miR-135a) to GATA-3 was also observed, and overexpression of this miRNA decreased GATA-3 mRNA and protein expression. However, the effects of miR-135a on MCs during AR are currently unknown. In the present study, we utilized a lentiviral (LV) vector to intranasally administer miR-135a to ovalbumin (OVA)-sensitized AR mice. Following miR-135a treatment, the total serum IgE concentration observed during AR was significantly reduced. In the nasal mucosa, the expression of T-box expressed in T cells (T-bet) was higher, whereas that of GATA-3 was lower in the AR mice following miRNA treatment. Notably, during AR, the ratio of type 1 T-helper cells (Th1) to type 2 (Th2) cells in the spleen is unbalanced, favoring Th2. However, administering miR-135a to the AR mice appeared to balance this ratio by increasing and decreasing the percentage of Th1 and Th2 cells, respectively. MiR-135a also appeared to strongly suppress the infiltration of eosinophils and MCs into the nasal mucosa, and it was specifically localized in the MCs, suggesting that its influence is modulated through regulation of GATA-3 in these cells. Additional work identifying the full therapeutic potential of miR-135a in the treatment of AR and diseases involving allergen-induced inflammation is warranted.

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.

Impulse noise was adopted in adult rats to built acute deafferent animal model. Differential proteomics techniques were applied to detect the changes of protein expression in the auditory cortex before and after the noise exposure.Thirty adult SD rats were divided into three groups: normal group, rats with acute noise exposure and rats 28 days recovery after noise exposure (n=10/group). All animals were exposed to impulse noise at 156 dB for 50 pulses with a rise-time of 100 µs and duration of around 0.25 ms. ABR was used to evaluate the auditory function. The two-dimensional gel electrophoresis (2-DE) and matrix-assisted laser desorption ionization time-of-flight mass spectrometer (MALDI-TOF-MS) were used to identified the differential protein expression.Compared with the normal group, ABR thresholds were found significantly increased at 2, 4, 8, 16, 32 kHz (P0.05) in the acute and recovery groups. There was a 40-60 dBSPL ABR threshold shift at all tested frequencies immediately after impulse noise exposure. There was a partial recovery of ABR thresholds at 7 day to 28 days after impulse noise exposure. In addition, it seemed that the thresholds were rather stable and no further ABR threshold recovery was observed from 14 day to 28 days after the impulse noise exposure. Using differential proteomic techniques, 36 spots containing 27 proteins were revealed and identified in auditory cortex. Those proteins are related to cytoskeleton, neurotransmission, energy supply, mitochondrial function and synaptic remolding.Impulse noise may influence the function of microtubule transport and cell metabolism, there after affect the neurotransmission of auditory neurons. The compensatory changes such as pre- and postsynaptic or such related functional changes may also happen in auditory cortex after the deafferentation treatment.

To observe the distribution of Gd-DTPA in the inner ear of guinea pig by MRI at different time points after intratympanic administration, explore the optical time for observing the whole inner ear. To study the pharmacokinetic feature of Gd-DTPA in the inner ear, and find out whether discrimination of endolymph and perilymph can be obtained under current conditions.Sixty-five guinea pigs were randomly divided into 13 groups, after diluted Gd-DTPA intratympanic injection, each group of guinea pigs were scanned through MRI (3D-T1 FSE sequence) at different time points (0 h, 0.5 h, 1 h, 2 h, 4 h, 6 h, 8 h, 10 h, 12 h, 24 h, 48 h, 72 h, 96 h). Pixel intensity values of some locations in the inner ear were analyzed using e-Film software, then pixel intensity was converted into concentration using the results of previous in-vitro study. ABR thresholds of bilateral ears before, 1 d and 7 d after intratympanic injection were compared.Six hours after transtympanic Gd-DTPA injection was the time point when contrast agent was distributed all over the inner ear, and reached a high concentration: 589.29, 552.54, 570.17, 255.08, 107.09 and 139.18 µmol/L in the vestibule, scala vestibuli and scala tympani of basal turn, the 2(nd), the 3(rd) and the apical turn individually, that was also the optimal time for observing the whole inner ear by MRI. Perilymph appeared to be preferentially enhanced relative to the endolymph, resulting in a distinction between the scales of the inner ear. There was no significant difference between the experimental ear (diluted GD-DTPA injected ear) and contrast ear (physiological saline injected ear) on 1 d and 7 d.The best time getting MRI imaging after intratympanic diluted GD-DTPA injection is 6 h. After diluted agent injection perilymph can be enhanced so as to be differentiated with endolymph by MRI, diluted agent have no obvious effect on the ABR threshold. The pharmacokinetic feature of Gd-DTPA in the inner can be studied using MRI.

To establish the activation of NF-kappaB in middle ear cholesteatoma, discuss the relationship of NF-kappaB and the gene expression of IL-6 and explore the pathogenesis of middle ear cholesteatoma.Ten cases of cholesteatoma and 6 cases of normal external meatal skin were obtained from middle ear surgery. The NF-kappaB DNA binding activity and the mRNA level of IL-6 in these two kinds of tissues were detected by electrophoretic motility shift assay (EMSA) and Reverse transcription polymerase chain reaction (RT-PCR) respectively. The relationship of NF-kappaB DNA binding activity and the mRNA level of IL-6 were analyzed.The NF-kappaB DNA binding activities of cholesteatoma [(15.9 +/- 8.2)%] were higher than those in normal skin [(1.36 +/- 0.94)%, t = 3.502, P0.05]. The expression of IL-6 mRNA was increased significantly in patients with cholesteatoma, as compared with that in the control specimens (t = 2.166, P0.05) and had a significant positive correlation with NF-kappaB DNA binding activity (r = 0.752, P0.05).The IL-6 mRNA expression in cholesteatoma is closely related with the activity of NF-kappaB. It is tempting to speculate that NF-kappaB play a key role in the activation of cytokine in cholesteatoma.

A retrospective review of seventeen patients who were operated through the maxillary swing approach was carried out to assess the efficacy of this approach in the management of tumors of the central and lateral cranial base.From May 2000 to January 2005, 17 patients with primary or recurrent neoplasms involving the central cranial or lateral base underwent surgical resection via maxillary swing approach. Ten patients were male, and other seven patients were female, and age range was 7 to 58 years, with a mean age of 42. 6 years. Eight patients had tumors originally involving lateral cranial base, and nine patients had tumors originated from central cranial base. The pathology spectrum was very wide. Among them, five suffered from chordoma, two had rhabdomyosarcoma, two had squamous cell carcinoma, one had malignant fibrous histiocytoma, one had malignant melanoma, one had esthesioneuroblastoma, one had invaded hypophysoma, two had schwannoma, one had pleomorphic adenoma, and one had angiofibroma. Three patients had received previous surgery, two patients had previous radiation therapy and nine patients received postoperative radiotherapy.Sixteen of all seventeen patients had oncologically complete resection, one had near-total resection. This group patients was followed up from 10 to 60 months, with a median follow-up time of 28 months. Two patients died 14 and 26 months after surgery respectively, as a result of local recurrence and metastasis. One patient defaulted follow-up at 12 months after operation, and the other 14 patients were alive at the time of analysis. Of the 12 malignant tumors, the 1-and 2-year survival rate were 91.67% and 72.92%, respectively. The facial wounds of all patients healed primarily, and there were no necrosis of the maxilla, damage of the temporal branch of the facial nerve, lower-lid ectropion, and facial deformity. Epiphora and facial hypoesthesia were detected in all patients. Four patients (23.5%) developed palatal fistula, ten patients developed serous otitis media (58.8%), and four patients developed a certain degree of trismus (23.5%). Cerebrospinal fluid leak occurred in two patients. They subsequently healed with conservative management.The maxillary swing approach is a proven method for access to the central and lateral skull base with good exposure and acceptable morbidity. Complications and sequelae associated with this approach include facial scarring, transaction of the infraorbital nerve, impaired lacrimal drainage, eustachian tube dysfunction and serous otitis, palatal fistula, trismus etc. Some procedures should be performed for reducing the incidence and severity of complications in the maxillary swing approach.

To establish a human laryngeal carcinoma cell line unassociated with human papillomavirus (HPV).Viable tissue of a well-differentiated laryngeal squamous cell carcinoma was obtained and tested negative for the presence of HPV by polymerase chain reaction. Minced tissue fragments were then transplanted subcutaneously into nude mice. After two successive passages, the tumor tissue was seeded into culture flasks and incubated in a medium containing 10% fetal bovine serum, insulin and epidermal growth factor. Tumor cell phenotype and molecular features were determined by various methods.A stable cell line, designated as Lscc-02, was successfully established after 86 culture passages. The cells grew as a monolayer with epithelioid features. The cell doubling time was approximately 39.1 hours. The human origin of the tumor cells was confirmed by karyotype analysis. The squamous epithelial phenotype was demonstrated by the immunopositivity of anti-cytokeratin antibodies and ultrastructural presence of tonofilaments and desmosomes. The malignant nature of the cells was documented by their clonal formation in soft-agar and tumorigenicity in nude mice. Lscc-02 cells expressed carcinoembryonic antigen (CEA) and were negative for HPV DNA.This newly established Lscc-02 laryngeal squamous cell carcinoma cell line may be a useful model for investigating laryngeal carcinoma unrelated to HPV infection, and the role of HPV in the progression of human laryngeal carcinoma.

To explore the effect and significance of endoscopic surgery for intranasal meningoencephaloceles via the supraorbital keyhole approach.From January 1999 to December 2001, seven cases of intranasal meningoencephaloceles were treated by mini-invasive endoscopic surgery via supraorbital keyhole approach. The operation was carried out through supraorbital keyhole craniotomy approximately 2.0-2.5 cm in diameter following a skin incision just in the eyebrow. The roughly 3-4 cm skin incision began medial to the supraorbital nerve and ended within the lateral edge of the eyebrow. After the frontal lobe was elevated and the skull base defects exposed, the hernia sac was cut off at bone defect and the lump was resected via nasal cavity. The bone defect were repaired using bone flap, self fat, fascia of rectus abdominis muscle and Ear-Brain Glue.Seven patients were successfully treated at initial operation without recurrence and cerebrospinal fluid leak during follow-up 6 months to 2 years after surgery.The supraorbital keyhole approach is a safe approach in the hands of experienced surgeons for the mini-invasive treatment of meningoencephaloceles.

To study the feasibility of alpha-cyanoacrylate medical adhesive in fixation of intratemporal facial nerve when nerve was repaired within chitin chamber, and to investigate the nerve regeneration.Nerve defect of 6 mm was made in left intratemporal facial nerves of 48 rabbits. All the defects were bridged with chitin chamber and were fixed by alpha-cyanoacrylate medical adhesive, surgical suture and natural union. Nerve function test and histomorphological examination were carried out at 1 month and 3 months after repair.It was observed that the nerve was fixed firmly to the chamber with no crack or crease by alpha-cyanoacrylate medical adhesive. The regenerated new nerve fibers were more regular and denser and the neurological function recovered much better in the group fixed by alpha-cyanoacrylate medical adhesive than in the groups those fixed by surgical suture and natural union.The medical adhesive is strong in adhesion and beneficial to nerve repair; repair of intratemporal facial nerve defect within chitin chamber fixed by alpha-cyanoacrylate medical adhesive is feasible, simple and timesaving.

Objective 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) on hTERT expression and telomerase activity in laryngeal cancer cells. Design Short hairpin RNA expression vectors targeting the messenger RNA of hTERT were constructed. Cells were treated with shRNA expression vectors directed against 2 different hTERT sites, control vectors that included mismatched shRNA and those without shRNA. The expression of hTERT was determined by reverse-transcriptase polymerase chain reaction and Western blotting. The activity of telomerase was measured by telomeric repeated amplification enzyme-linked immunosorbent assay. The cell viability was examined using the 3-(4,5-dimethyl thizol-2-yl) 2,5-diphenyl tetrazolium bromide assay. Results We found that treatment of shRNA expression vectors induced a significant decrease in hTERT messenger RNA expression, the level of hTERT protein, telomerase activity, and cell viability. All of these effects were seen regardless of the target site, and the shRNA control showed none of these effects. Conclusion Our results suggest that shRNA directed against hTERT inhibits telomerase activity through suppression of the hTERT expression in laryngeal cancer cells and that RNA interfering technology may be a promising strategy for the treatment of laryngeal cancers.