gic rhinitis animal model Al(OH)3 colloid gel of different concentration (0.5-5 mg) may cause side effects such as foreign body granuloma.

Objective To investigate the histopathological changes in cricoarytenoid joints in 32 animal models.The characteristic histopathological changes of arytenoid cartilages after recurrent nerve paralysis were evaluated. Methods Sixteen dogs (32 vocal folds, 8 as normal control) were divided into different animal models of recurrent nerve paralysis as transection, haft-section, ligation, or crush. The histopathological finds of arytenoid cartilages were analysed.Results Arytenoid cartilages showed fibrin (12/24), disruption of the fibrous membrane(9/24), fibrillation (7/24) and degenerative changes in their joint surface structure (3/24) at various levels of intensity.The fibrin and disruption of the fibrous membrane were found 1 month after injury, and all changes appeared in 6 months. The fibrillation and arytenoid cartilages degenerative changes revealed in transaction group and ligation group, and became stronger in time of 6 months. The correlation among the fibrillation ratio and the normal control was positive ( t were 6.23 and 3.65, P<0.01 ). The correlation among the number of cellular of arytenoid cartilages and the normal control was positive (t= 2.78, P < 0. 05 ). The fibrillation (7) and arytenoid cartilages degenerative changes (3) revealed in vocal fold fixation to influence the recovery of laryngeal function.Conclusions The histopathological change of cricoarytenoid joint after recurrent nerve paralysis was related to the severity of neural injury. Influence the recovery of laryngeal function more often from 6 months.

BACKGROUNDGene therapy and epigenetic therapy have gained more attention in cancer treatment. However, the effect of a combined treatment of gene therapy and epigenetic therapy on head and neck squamous cell carcinoma have not been studied yet. To study the mechanism and clinical application, human laryngeal carcinoma cell (Hep-2) tumor-bearing mice were used.

METHODSA xenograft tumor model was established by the subcutaneous inoculation of Hep-2 cells in the right armpit of BALB/c nu/nu mice. The mice with well-formed tumor were randomly divided into six groups. Multisite injections of rAd-p53 and/or 5-aza-dC were used to treat tumor. Tumor growth was monitored by measuring tumor volume and growth rate. p53 and E-cadherin protein levels in tumor tissues were detected by immunohistochemical staining. The mRNA levels were monitored with FQ-PCR.

RESULTSGene therapy was much more effective than single epigenetic therapy and combined therapy. The gene therapy group has the lowest tumor growth rate and the highest expression levels of p53 and E-cadherin.

CONCLUSIONSThe combined treatment of gene and epigenetic therapy is not suggested for treating head and neck carcinoma, because gene therapy shows an antagonistic effect to epigenetic therapy. However, the mechanisms of action are still unclear.

Animals ; Azacitidine ; analogs & derivatives ; therapeutic use ; Cadherins ; analysis ; DNA Modification Methylases ; antagonists & inhibitors ; Epigenesis, Genetic ; Genes, p53 ; Genetic Therapy ; Humans ; Laryngeal Neoplasms ; genetics ; pathology ; therapy ; Male ; Mice ; Mice, Inbred BALB C ; Tumor Suppressor Protein p53 ; analysis ; Xenograft Model Antitumor Assays

BACKGROUNDCurrently anti-inflammatory therapy with steroids for allergic rhinitis need long-term repeated administration, although it is effective. Gene therapy is being suggested to substitute it. The aim of this study was to investigate nonviral vector mediated exogenous gene expression in COS-7 cells in vitro and the effect of intranasal mouse interleukin (mIL)-12 transgene expression on allergen induced eosinophil infiltration of nasal mucosa in a murine model of allergic rhinitis.

METHODSIn vitro COS-7 cells were infected with Epstein-Barr virus (EBV)/lipoplex. The expression of IL-12 p70 in cell culture supernatant was examined by enzyme-linked immunosorbent assay (ELISA). In mice with ovalbumin (OVA) induced allergic rhinitis, EBV/lipoplex was administered by nasal drops before OVA challenge once a day from day 1 to day 10. The expression of IL-12 mRNA and protein, the change of eosinophil count in nasal mucosa and serum total IgE were measured 24 hours after the last challenge.

RESULTSEBV/lipoplex could effectively transfect COS-7 cells. The expression of IL-12 p70 in cell culture supernatant was significantly more than in blank control. IL-12 via EBV plasmid vector transduction could be overexpressed in vivo. In pGEG.mIL-12 treated models, the nasal mucosa revealed a high level of widespread mIL-12 transduction by immunohistochemistry and in situ hybridization. Histological evaluation revealed marked suppression of eosinophil infiltration in nasal mucosa. The eosinophil count in allergic rhinitis group [(26.5 +/- 9.8)/high-power field (HPF)] was significantly increased over control group [(0.40 +/- 0.52)/HPF] (F = 56.94, P < 0.01), while the count in IL-12 gene therapy group [(4.60 +/- 2.63)/HPF] was significantly less than that of allergic group (F = 56.9, P < 0.01). Serum total IgE between in gene therapy mice [(88.83 +/- 6.71) ng/ml] and allergic rhinitis mice [(103.1 +/- 5.7) ng/ml] showed a significant difference (F = 1216, P < 0.05).

CONCLUSIONSNonviral EBV plasmid vector, pGEG.mIL-12 was able to overexpress exogenous gene both in vitro and in murine nasal mucosa in vivo. IL-12 overexpression via EBV/lipoplex could stem allergen induced eosinophil infiltration in nasal mucosa in murine models of allergic rhinitis, which may suggest a new cytokine immunogenetic therapy for allergic rhinitis.

Administration, Intranasal ; Animals ; COS Cells ; Cercopithecus aethiops ; Eosinophilia ; therapy ; Genetic Therapy ; Herpesvirus 4, Human ; genetics ; Immunoglobulin E ; blood ; Interleukin-12 ; genetics ; Lipids ; administration & dosage ; Male ; Mice ; Mice, Inbred BALB C ; Nasal Mucosa ; metabolism ; Rhinitis, Allergic, Perennial ; therapy ; Rhinitis, Allergic, Seasonal ; therapy

Objective To evaluate the effect of intranasal liposome-mediated IL-12 gene therapy on the eosinophils and IL-5 in the murine model of allergic rhinitis. Methods Thirty-six BALB/C mice were randomly divided into allergic rhinitis (AR) group, gene therapy group and control group. Allergic rhinitis group were sensitized and stimulated by ovalbumin(OVA), and gene therapy group were administered with hposome-mediated pGEG. m IL-12 transnasally before stimulated. The eosinophiis in bone marrow were counted by Wright's staining, and the eosinophils in nasal mucosa were counted by HE staining. The eosinophils of peripheral blood were detected by flow cytometry. The expression of IL-5 in bone marrow and nasal mucosa was examined by immunohistochemistry. The IL-5 in serum was detected by ELISA. Results Among the three groups, the difference of all data was statistically significant (P<0.01). Multiple Comparison showed that the ratio of eosinophils to white cells and the mount of IL-5 positive cells in nasal mucosa and bone marrow of gene therapy group was significantly lower than that of AR group (P<0.05).The ratio of eosinophils to granulocyte(0.124±0.031) and the expression level of IL-5[(29. 51±6. 68) pg /ml]in peripheral blood [0.184±0. 079 and (56. 58±16. 80) pg/ml] were significantly lower in gene therapy group than in AR group (P<0.05). Conclusions Transnasal administration of liposome- mediated pGEG. mIL-12 could depress the expression of IL-5 in bone marrow, peripheral blood, and nasal mucosa, to influence the proliferation and differentiation of eosinophils and decrease the delivery and transference of eosinophils to peripheral blood and nasal mucosa. It may be a new treatment for respiratory tract allergic inflammation.

Objective To investigate the treatment for injured vocal folds by implanting autologous adipose-derived mesenchymal stem cells ( ADSC ), and to observe the characteristics of lamina propria of the vocal folds and its major extracellular matrix ( ECM ), as well as the growth features of ADSC. Methods The lamina propria was injured by localized resection in fifty-three vocal folds of thirty-four rabbits.Isolation, culture and identification of ADSC were performed in twenty rabbits. Three to five days after vocal folds injury, autogeneic ADSC were implanted into the injured vocal folds. As control, scaffolds ( collagen or hyaluronic acid) and none were injected into eighteen and fifteen vocal folds respectively. Larynges were harvested at 15 days, 40 days and 3, 6, 12 months after operation. The growth and distribution of ADSC were detected by DAPI stain. Meanwhile, HE staining was performed for histopathologic research, Masson trichrome staining, Alcian Blue staining and immunohistochemical staining were used for collagen,hyaluronic acid and fibronection respectively. Results ADSC showed a spindle-shaped adherent growth,with multi-differentiation potential. After implanting into the injured vocal fold, ADSC can survive in vocal fold lamina propria. In ADSC implanting group, the morphology and histologic structure of vocal folds were similar to normal in six and twelve months after ADSC implanting respectively. collagen had an increasing trend in three months, with the disorderly distribution in the vocal fold lamina propria, then it became decreasing until the twelveth month, when concentration closed to normal, however, distribution of collagen was a little irregular. The content of hyaluronic acid increased within forty days and distributed in the lamina propria, then gradually reduced to normal levels in the following twelve months, and limited in the superficial and middle layers of lamina propria, which closed to normal. Fibronectin in the lamina propria continued to scattered at the peak levels at fortieth day, then decreased in the later twelve months when the content became near normal. In the untreated group, vocal fold showed a local scar contracture at third month after injury, histology showed large number of fibrous tissue ( mainly collagen fibers) hyperplasia with a tendency of increasing, disorders was also found in vocal fold lamina propria. In the scaffold implanting group, the changes were between the two groups above. Conclusions ADSC are good source of seed cells for vocal fold tissue engineering, which have the ability for promoting injured rabbit vocal folds ECM secretion, rational distribution and part ordering arrangement. ADSC also play an important role in vocal folds reparation and regeneration.

Objective To analyze the correlation between airborne pollen concentrations and symptoms in patients with pollen allergic rhinitis.Methods Durhum sampler was used to collect the pollen concentration and species from June to September in 2011.The clinical skin prick test (SPT) data were analyzed.The patients with pollen allergic rhinitis were divided into pure pollen allergic rhinitis group (pollen group) and pollen combined perennial allergens allergic rhinitis group (combined group).Symptom scores of patients were assessed,and correlation between pollen concentration and onset of symptoms of patients were analyzed.SPSS 16.0 software was used to analyse the data.Results While the peak of Summer-Autumn pollen concentration appeared from August 20 to September 15,the major pollen included Artemisia L,Chenopodium album and Humulus scandens.The peak of pollen concentration in one day reached 638/1000 mm2.The patients taken SPT from June to September accounted for 51.9％ of the patients in whole year,among which SPT pollen positive patients were 1509,60.7％ of all SPT positive patients.The amount and rate of SPT positive patients showed significant correlation with pollen concentration( r value were 0.90 and 0.99,both P ＜0.05 ).Onset of symptoms in two groups was correlated with pollen concentration in Summer-Autumn.Symptoms of cough in combined group showed more severe compared with patients with pollen group (t =2.36,P ＜ 0.05 ).Conclusions Pollen concentration has a major effect on onset of symptoms of allergic rhinitis.Airborne pollen monitoring has important preventive and therapeutic significance on patients with allergic rhinitis.

Objective To observe the histopathological finding of bone remodeling in rabbit sinusitis model at different time and the tendency, and to discuss the effect of bone in the pathogenesis of sinusitis. Methods First, the rabbit sinusitis model was made, then the experimental animals were divided into 3 groups according to the time of infection. There were 8 rabbits in each experimental group, and 4 rabbits in the control group. The sinus specimen were collected, embedded and stained. The bone in the inoculating side and noninoculating side was scored, and the bone in inoculating side was evaluated quantitatively and semiquantitatively. The parameters included the thickness of mucesa, mucoperiosteum, the density of osteoblast, the amount of esteoclast. Results The average bone score in the inoculating side was 2. 250,2.875,2.875 ; in the noninoculating side was 1. 625,2.250,2.500. Between group A and the control group, the difference of all three parameters had statistical significance. Between group B and group A, the difference of the thickness of mucosa and the density of osteoblast had statistical significance. Between group C and group B, none of the three parameters had statistical significance. Conclusions Bacterial sinusitis can lead to bone remodeling, obvious bone destroy can occur at the early phase, then the bone proliferation follows. These results demonstrate that bone remodeling is one of the basic histopathological characters of CRS and might be the reason to lead CRS to a constant and chronic process of inflammation.

OBJECTIVETo analyze the impact of olfactory nerve transection on the apoptosis of mice olfactory receptor neurons (ORN), and discuss the reliability of this experimental model.

METHODSAfter olfactory nerve transection of mice, anterograde horseradish peroxidase (HRP) tracing was performed to confirm the completion of nerve transection. On 8 h, 2 d, 3 d and 5 d after surgery, TdT mediated deoxyuridine triphosphate-biotin nick end labelling (TUNEL) was used to observe the apoptosis of ORN, while relative semi-quantitative RT-PCR and immunohistochemistry were used to reflect the expression of olfactory marker protein (OMP, special marker of mature ORN) in olfactory epithelium.

RESULTSNo HRP label was observed in olfactory bulb after olfactory nerve transaction. Both TUNEL-positive and OMP-positive cells were ORN. After the surgery, TUNEL-positive cells increased remarkably and peaked on 2 d after the surgery. Meanwhile OMP mRNA in olfactory epithelium began to decrease markedly till 5 d after the surgery, and the olfactory epithelium got thinner accordingly.

CONCLUSIONSThis experimental model can be used reliably to sever mice olfactory nerve and manipulate simultaneous apoptosis of mice ORN.

Animals ; Apoptosis ; Denervation ; Mice ; Mice, Inbred C57BL ; Olfactory Nerve ; cytology ; metabolism ; surgery ; Olfactory Receptor Neurons ; metabolism ; pathology

OBJECTIVETo investigate whether the local application of IL-12 gene with EBV-plasmid vector to nasal mucosa could prevent allergic inflammation in murine allergic rhinitis model.

METHODSThirty-six BALB/C mice were randomly divided into allergic rhinitis group gene therapy group and control group. In mice with OVA-induced allergic rhinitis, the EBV/lipoplex (a novel cationic lipid combined with EBV-plasmid vector, pGEG. mIL-12) was locally administered into nasal mucosa before OVA challenge. The expression of IL-12 mRNA and protein, the change of eosinophilia and mast cell, and Th2 cytokine production in the nasal mucosa were measured.

RESULTSThe amounts of IL-12 mRNA positive cells and IL-12 positive cells in nasal mucosa of gene therapy group were significantly higher than that of allergic rhinitis group (P < 0.01 and P < 0.05). The amount of eosinophils, mast cells, and the level of IL-5 expression in nasal mucosa in allergic rhinitis group were significantly higher than those in gene therapy group and control group (P < 0.01). The level of total IgE of peripheral blood in allergic rhinitis group was significantly higher than that in gene therapy group and control group (F = 1216.21, P < 0.01).

CONCLUSIONSThese findings indicated that IL-12 mRNA and protein were expressed effectively after the local administration of pGEG. mIL-12 in the nasal mucosa. The local application of pGEG. mIL-12 is effective in modulating nasal allergic response and may be a convenient method for future approach to allergic rhinitis.

Animals ; Genetic Therapy ; Genetic Vectors ; Interleukin-12 ; genetics ; therapeutic use ; Male ; Mice ; Mice, Inbred BALB C ; Nasal Mucosa ; metabolism ; Rhinitis, Allergic, Perennial ; therapy