Objective To prepare isophenylcyclopentylamine hydrochloride capsules and evaluate its quality. Methods The suitable excipients were selected by the drug-excipient compatibility test,and after the formulation and preparation process design and screening,the micromeritic property and hygroscopicity of contents of capsules as well as the basic performance and stability of is-ophenylcyclopentylamine hydrochloride capsules were evaluate. Results The selected formulation was composed of isophenylcyclo-pentylamine hydrochloride 10 mg,anhydrous dibasic calcium phosphate 128.7 mg,mannitol 128.7 mg and aerosil 2.7 mg.The angle of repose of intermediate powders was(30.71±1.09)°,aerated and packed bulk densities were(0.76±0.01)and(0.90±0.02)g/ml re-spectively,Hausner value was 1.19±0.01,Carr′s index was(15.86±1.05)%,indicating good flowability and filling property as well as relatively low humidity.The contents,uniformity of contents and dissolution of capsules fulfilled the requirements.The quality of ca-pusles kept well in the 6-month accelerated and long-term stability test.Conclusion The formulation is simple and reasonable,and the preparation process showed a good reproducibility,which might be suitable for industrialization.

BACKGROUNDAdenoid 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.

METHODSNine 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.

RESULTSThe 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.

CONCLUSIONSDown-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.

Adenoids ; cytology ; Antimicrobial Cationic Peptides ; metabolism ; Child ; Epithelial Cells ; metabolism ; Female ; Humans ; Immunity, Innate ; genetics ; physiology ; Male ; Receptors, Pattern Recognition ; metabolism ; Sinusitis ; metabolism ; Toll-Like Receptors ; metabolism

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

BACKGROUNDBiofilms 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).

METHODSA 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.

RESULTSThirteen (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.

CONCLUSIONOur 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.

Adolescent ; Adult ; Biofilms ; Chronic Disease ; Female ; Humans ; Male ; Microscopy, Electron, Scanning ; Middle Aged ; Nasal Mucosa ; pathology ; Nasal Polyps ; etiology ; microbiology ; pathology ; Rhinitis ; etiology ; microbiology ; pathology ; Sinusitis ; etiology ; microbiology ; pathology

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 effects of gene therapy and epigenetic therapy on the tumor growth of laryngeal carcinoma and the underlying mechanisms.Methods 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.Results By the day 20 of the treatments,the mean tumor volumes were (106.09±24.40)mm3 in p53 therapy group,(166.55 ± 40.11 )mm3 in epigenetic therapy group,(126.11 ±22.49 )mm3 in combination therapy group,and (252.83 ± 54.09 )mm3 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 epigenetie 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 ).Conclusions Xenografled tumors of human laryngeal carcinoma cells are inhibited by gene therapy,the epigenetic therapy and the ombination 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.

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.

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 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 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.