Expression of Innate Immunity Genes in Epithelial Cells of Hypertrophic Adenoids with and without Pediatric Chronic Rhinosinusitis: A Preliminary Report / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Adenoid hypertrophy (AH) is associated with pediatric chronic rhinosinusitis (pCRS), but its role in the inflammatory process of pCRS is unclear. It is thought that innate immunity gene expression is disrupted in the epithelium of patients with chronic rhinosinusitis (CRS), including antimicrobial peptides and pattern recognition receptors (PRRs). The aim of this preliminary study was to detect the expression of innate immunity genes in epithelial cells of hypertrophic adenoids with and without pCRS to better understand their role in pCRS.</p><p><b>METHODS</b>Nine pCRS patients and nine simple AH patients undergoing adenoidectomy were recruited for the study. Adenoidal epithelium was isolated, and real-time quantitative polymerase chain reaction (RT-qPCR) was employed to measure relative expression levels of the following messenger RNAs in hypertrophic adenoid epithelial cells of pediatric patients with and without CRS: Human β-defensin (HBD) 2 and 3, surfactant protein (SP)-A and D, toll-like receptors 1-10, nucleotide-binding oligomerization domain (NOD)-like receptors NOD 1, NOD 2, and NACHT, LRR and PYD domains-containing protein 3, retinoic acid-induced gene 1, melanoma differentiation-associated gene 5, and nuclear factor-κB (NF-κB). RT-qPCR data from two groups were analyzed by independent sample t-tests and Mann-Whitney U-tests.</p><p><b>RESULTS</b>The relative expression of SP-D in adenoidal epithelium of pCRS group was significantly lower than that in AH group (pCRS 0.73 ± 0.10 vs. AH 1.21 ± 0.15; P = 0.0173, t = 2.654). The relative expression levels of all tested PRRs and NF-κB, as well as HBD-2, HBD-3, and SP-A, showed no statistically significant differences in isolated adenoidal epithelium between pCRS group and AH group.</p><p><b>CONCLUSIONS</b>Down-regulated SP-D levels in adenoidal epithelium may contribute to the development of pCRS. PRRs, however, are unlikely to play a significant role in the inflammatory process of pCRS.</p>

Antagonism between gene therapy and epigenetic therapy on human laryngeal carcinoma tumor-bearing mice / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Gene 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.</p><p><b>METHODS</b>A 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.</p><p><b>RESULTS</b>Gene 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.</p><p><b>CONCLUSIONS</b>The 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.</p>

Correlation between symptoms of pollen allergic rhinitis and pollen grain spreading in summer and autumn / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To analyze the correlation between airborne pollen concentrations and symptoms in patients with pollen allergic rhinitis.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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 mm(2). 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).</p><p><b>CONCLUSIONS</b>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.</p>

Inhibition of human laryngeal carcinoma growth by gene therapy and epigenetic therapy / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To observe the effects of gene therapy and epigenetic therapy on the tumor growth of laryngeal carcinoma and the underlying mechanisms.</p><p><b>METHODS</b>The animal model of human laryngeal carcinoma was established by the subcutaneous inoculation of Hep-2 cells at the right armpit of BALB/c nu/nu mice. The tumor-bearing mice were randomized into 4 groups, p53 therapy group(rAd-p53), epigenetic therapy group(5-aza-dC), combination therapy group (rAd-p53+5-aza-dC) and control group. The gene and protein expressions of molecular markers p53 and E-cadherin were detected by FQ-PCR and immunohistochemistry.</p><p><b>RESULTS</b>By the day 20 of the treatments, the mean tumor volumes were(106.09 ± 24.40)mm(3) in p53 therapy group, (166.55 ± 40.11) mm(3) in epigenetic therapy group, (126.11 ± 22.49) mm(3) in combination therapy group,and (252.83 ± 54.09) mm(3) in control group. Both gene therapy (F = 37.30, P < 0.05) and epigenetic therapy (F = 4.79, P < 0.05) inhibited the growth of xenografted tumors, with an interaction effect (F = 22.01, P < 0.05) between the two groups. The integral optical density value of p53 protein expression of p53 therapy group (628.07 ± 95.16) was significantly higher than that of combination therapy group (494.76 ± 100.22), (t = 8.72, P < 0.05). The integral optical density values of E-cadherin protein expression were 558.89 ± 97.58 in p53 therapy group, 380.41 ± 90.60 in epigenetic therapy group, 494.76 ± 102.88 in combination therapy group,and 162.60 ± 40.38 in control group respectively, indicating the enhancements of E-cadherin protein expression by gene therapy (F = 45.24, P < 0.05) or epigenetic therapy(F = 5.73, P < 0.05)and the existence of interaction effect (F = 21.82, P < 0.05) between gene therapy and epigenetic therapy. The expression levels of p53 gene were 4.43 ± 0.12 in p53 therapy group, 1.06 ± 0.11 in epigenetic therapy group, 3.51 ± 0.10 in combination therapy group,and 1.09 ± 0.11 in control group, respectively, showing an interaction effect between gene therapy and epigenetic therapy (F = 298.11, P < 0.05). The expression levels of E-cadherin gene were 4.50 ± 0.34 in p53 therapy group, 2.02 ± 0.16 in epigenetic therapy group, 2.99 ± 0.12 in combination therapy group, and 1.00 ± 0.11 in control group, respectively. The expression of E-cadherin gene was enhanced by gene therapy (F = 329.12, P < 0.05)or epigenetic therapy(F = 88.57, P < 0.05), with an interaction effect between the two therapies (F = 122.17, P < 0.05).</p><p><b>CONCLUSIONS</b>Xenografted tumors of human laryngeal carcinoma cells are inhibited by gene therapy, the epigenetic therapy and the combination therapy. The gene therapy was significantly better than the epigenetic therapy or the combination therapy. There might be antagonistic effect between p53 and 5-aza-dC.</p>

Clinical and histopathologic features of biofilm-associated chronic rhinosinusitis with nasal polyps in Chinese patients / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Biofilms have given new insights to the understanding of pathogenesis of chronic rhinosinusitis (CRS). However, the link between biofilms formation and local inflammatory response remains poorly defined in CRS with nasal polys. The aim of this study was to determine the potential association of the presence of biofilms in the nasal mucosal tissues with clinical features in Chinese patients, which had CRS with nasal polyps (CRSwNP).</p><p><b>METHODS</b>A total of 19 patients with CRSwNP and 12 patients with non-CRS were subjected to endoscopic surgery and their nasal mucosal tissue specimens were examined histologically and by scanning electron microscopy (SEM). Their demographic and clinical features were recorded.</p><p><b>RESULTS</b>Thirteen (68.4%) out of the 19 specimens from patients with CRSwNP, but none from control patients, were positive for biofilms that displayed typical characteristics of bacterial and fugal structures. The presence of biofilms in the nasal mucosal tissues was associated with significantly greater values of purulent nasal discharge and preoperative Lund-Kennedy scores, higher levels of serum total IgE and percentages of subjects with endoscopic surgery (ESS) history in patients with CRSwNP, and more severe inflammation in the nasal mucosal tissues of patients with CRSwNP.</p><p><b>CONCLUSION</b>Our study demonstrated the presence of biofilms in the nasal mucosal tissues of many patients, contributing to the understanding of the pathogenic process of CRSwNP in Chinese patients.</p>

Characterization of vocal fold regeneration after adipose-derived mesenchymal stem cells implanting / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>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.</p><p><b>METHODS</b>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 fibronectin respectively.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSIONS</b>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.</p>

Histopathological change of cricoarytenoid joint after laryngeal recurrent nerve paralysis in dogs / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>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.</p><p><b>METHODS</b>Sixteen dogs (32 vocal folds, 8 as normal control) were divided into different animal models of recurrent nerve paralysis as transection, half-section, ligation, or crush. The histopathological finds of arytenoid cartilages were analysed.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSIONS</b>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.</p>

Effect of intranasal IL-12 gene therapy on the mice eosinophils and IL-5 in the murine model of allergic rhinitis / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>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.</p><p><b>METHODS</b>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 liposome-mediated pGEG. mIL-12 transnasally before stimulated. The eosinophils 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.</p><p><b>RESULTS</b>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).</p><p><b>CONCLUSIONS</b>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.</p>

Histopathological observation of bone remodeling in rabbit sinusitis model / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>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.</p><p><b>METHODS</b>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 mucosa, mucoperiosteum, the density of osteoblast, the amount of osteoclast.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSIONS</b>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.</p>

Evaluation of the safety of aluminium adjuvant in the preparation of allergic rhinitis animal model / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To evaluate the effect and safety of the formulation and dosage of aluminium adjuvant, Al(OH)(3), in the preparation of allergic rhinitis animal model.</p><p><b>METHODS</b>Sixty health BALB/c mice were divided randomly into 6 groups. Al(OH)(3) powder (5 mg) was used in one group, Al(OH)(3) colloid gel of different concentration (0.5 - 5 mg) was used in four groups, and normal saline was used in the control group. Ovalbumin injection and nasal topical challenge were used in the 5 testing groups to induce allergic rhinitis in mice. Normal saline was used in the control group.</p><p><b>RESULTS</b>Typical allergic rhinitis symptoms including frequent nasal scratching, and edema of peri-nasal mucosa were found in mice of the 5 mg Al(OH)(3) powder group. Eosinophils accumulation, goblet cells hyperplasia and hypersecretion were found in the mucosa of lateral nasal wall and inferior nasal turbinate. Neither obvious allergic rhinitis symptom, nor eosinophils accumulation in nasal mucosa was observed in the Al(OH)(3) colloid gel groups. Hemorrhagic ascites and lots of white nodules (foreign body granuloma) formation were found in the liver, spleen, and kidney of all mice of the 5 mg Al(OH)(3) colloid gel group. Five out of 10 mice of the 2 mg Al(OH)(3) colloid gel group exhibited above signs but of lower grade. Despite dispersed fine white sediment in the liver and mesentery, no obvious ascites was found in mice of the 1 mg and 0.5 mg Al(OH)(3) colloid gel groups.</p><p><b>CONCLUSIONS</b>Al(OH)(3) powder, 5 mg, is effective and safe in the preparation of allergic rhinitis animal model. Al(OH)(3) colloid gel of different concentration (0.5 - 5 mg) may cause side effects such as foreign body granuloma.</p>

Expression of odorant receptor genes on the olfactory epithelium following olfactory nerve disconnection / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To constitute the animal model of unilateral olfactory nerve transection and observe the expression level and distribution of odorant receptors.</p><p><b>METHODS</b>Thirty-two rats were divided into two groups: the olfactory nerve transection group (20) and the control group (12). The former group received the operation to transect the left olfactory nerve following the left olfactory bulb was exposed under microscope and the latter group did not give any disposal. At every stage of five days, two weeks, four weeks and six weeks after the operation, five rats from the nerve transection group and three from the control group were anaesthetized simultaneously, and olfactory epithelium were taken out after transcardial perfusion, then paraffin imbedding. Coronal sections were sliced for HE staining to observe the thickness changes of the olfactory epithelium, and for in situ hybridization (ISHs) to investigate the expression of olfactory receptor genes (Olr287, Olr226, Olr1493 and Olr1654) in the epithelium, also to evaluate the changes of the expression level and location of the selected receptors during the regeneration of olfactory epithelium.</p><p><b>RESULTS</b>HE staining showed that 5 days after the operation cell quantity and thickness of the olfactory epithelium decreased obviously, which increased gradually 2 or 4 weeks after operation. After 6 weeks' recovery, the thickness of the epithelium could reach the control level. The pattern of cell staining by ISH showed a specific spatial distribution along the anteroposterior (AP) and dorsoventral (DV) axis. Evidence suggested that odorant receptors were distributed in continuous and multiple overlapping bands in the normal or nerve transected-recovered epithelium rather than in the conventionally accepted three or four zones. The data also demonstrated that the distribution of sensory neuron types, as identified and defined by odorant receptor expression, was restored to normal or nearly so by 6 weeks after operation. Likewise, the numbers of probe-labeled neurons in the nerve transected-recovered had an obvious decrease 5 days after olfactory nerve transection. Reactive cells (x(-) +/- s) of Olr1493 in the operated side was (53.9 +/- 19.9), compared with (419.0 +/- 21.2) in the unoperated side, there was statistic significance between them (t = 63.960, P < 0.01). Reactive cells increased gradually according to the regeneration of the epithelium, and were nearly equivalent to the normal side 6 weeks later without significant differentiation (t = 2.600, P > 0.05), according to the absolute positive cells in the operated and unoperated side of (417.8 +/- 32.4) and (445.3 +/- 10.0) respectively.</p><p><b>CONCLUSION</b>The regeneration of the sensory neurons and receptors, both the number and the distribution, can recover to normal after olfactory nerve transection.</p>

Histolopathologic features of CT scan typing of chronic rhinosinusitis and relation with prognosis / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To evaluate the histopathologic changes of the ethmoid bone in CT scan typing of chronic rhinosinusitis (CRS), and investigate relations with outcomes of endoscopic surgery.</p><p><b>METHODS</b>One hundred and twelve random CRS patients undergoing endoscopic sinus surgery at Tongren hospital were prospectively evaluated. According to the preoperative CT scan, these patients were divided into three groups of CT scan typing. The bony changes and bone remodeling of ethmoid bone were graded and relations with outcomes of endoscopic surgery were investigated.</p><p><b>RESULTS</b>The CT scan typing results of ethmoid sinus of CRS were as followed: 22.3% (type III), 39.3% (type II), and 38.4% (type I). Comparison of bone remodeling and postoperative endoscopic scores between the three groups revealed a significant difference (P = 0.01, P = 0.02). A trend toward more advanced bone remodeling grade in association with a higher postoperative endoscopic scores was identified, with a statistically significant correlation.</p><p><b>CONCLUSION</b>The Chinese CT scan typing of CRS has its pathological basis and can a useful method to predict the surgical outcomes of CRS. The new bone formation or bony tissue remodeling of sinus often represents pathogenesis of CRS and implicates the prognosis of sinus surgery.</p>

Intranasal application of Epstein-Barr virus/lipoplex to abrogate eosinophillia in murine model of allergic rhinitis / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Currently 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.</p><p><b>METHODS</b>In 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.</p><p><b>RESULTS</b>EBV/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).</p><p><b>CONCLUSIONS</b>Nonviral 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.</p>

Impact of olfactory nerve transection on the apoptosis of mice olfactory receptor neurons / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To analyze the impact of olfactory nerve transection on the apoptosis of mice olfactory receptor neurons (ORN), and discuss the reliability of this experimental model.</p><p><b>METHODS</b>After 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.</p><p><b>RESULTS</b>No 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.</p><p><b>CONCLUSIONS</b>This experimental model can be used reliably to sever mice olfactory nerve and manipulate simultaneous apoptosis of mice ORN.</p>

Effect of intranasal interleukin-12 gene therapy for allergic rhinitis in murine model / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To 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.</p><p><b>METHODS</b>Thirty-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.</p><p><b>RESULTS</b>The 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).</p><p><b>CONCLUSIONS</b>These 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.</p>