This study was designed to explore the immune reconstitution efficacy of human thymic slices transplanted into renal capsule,subcutaneous or muscle of nude mice,and further explore the optimal location of heterotopic transplantation.The thymus tissue discarded from congenital heart disease patients was made into 0.5-1 mm thick tissue sections and cultured in vitro to remove immune cells.H&E staining and immunohistochemical staining were used to assess the residual tissue structure and cell types in thymic slices,while quantitative PCR methods were used to assess the function of residual cells in thymic slices.Then thymic slices were transplanted into the renal capsule,subcutaneous or muscle of nude mice,and the immune reconstitution efficacy was compared by flow cytometry and histology.Data showed that after 14 days of culture in vitro,the clearance rate of T lymphocytes in the thymic slices was more than 90%,and the epithelial cell network structure of the tissue was intact,while a large number of macrophages,dendritic cells and endothelial cells remained.Quantitative PCR results showed that the gene expression levels of epithelial cell markers and secreted cytokines in cultured thymic slices could be effectively maintained.Flow cytometry showed that at 16 weeks after transplantation,the proportion of T cells in peripheral blood of mice in different transplantation groups were significantly increased,whereas the proportion of T cells in muscle group was the highest.In situ histological examination showed that the regeneration of thymus tissue was detected at all three transplant sites.In addition,the graft detection rate was 40%in the renal capsule group,60%in the subcutaneous group and 100%in the musclegroup.In conclusion,the human thymic slices cultured in vitro for 14 days retain a complete thymic matrix microenvironment.Transplantation of human thymic slices can effectively reconstruct the ratio of T cells in nude mice,and the muscle is the most effective transplantation site.

Objective To evaluate the quality of different parts of agarwood formed by stimulation with biological induction method, thus to provide evidence for agarwood quality control. Methods The identification of agarwood and the determination of alcohol extracts from agarwood were achieved by the methods of Chinese Pharmacopeia. The determination of total chromones from agarwood was achieved by ultraviolet spectrophotometry(UVS), with aquilarone E as reference substance and 760 nm as the detective wavelength. Results The UVS method was stable within 5 h and the recovery rate was 95.71%(sR=2.09%). The average alcohol extract content of the outside part of the xylem of Aquilaria sinensis(Lour.) Gilg(part B) was 109.4 mg/g,which was higher than that (100 mg/g) of the Chinese Pharmacopoeia standard. The average content of total chromones was 15.73 mg/g. The rest identification results were accorded with the requirements of Chinese Pharmacopeia. The average alcohol extract content of the inside part of the xylem(part Z) was 47.6 mg/g, and the average content of total chromones was 2.66 mg/g. The rest identification results did not reach the requirements of Chinese Pharmacopeia. Conclusion The obtained part B of the agarwood meets the requirements of Chinese Pharmacopeia, while part Z has color changed and its quality has not yet reached the requirements. The results indicated that the biological induction method impacts Aquilaria sinensis xylem from outside to inside, and then the xylem grows into agarwood gradually.

Objective To establish a method for fingerprint analysis of amino acids from honey by high performance liquid chromatography ( HPLC). Methods Amino acids of honey were concentrated by 732 cation exchange resin, and then were treated by pre-column derivatization with phenyl isothiocyanate, with praline as control peak. The chromatography was performed on a Waters Symmetry C18 ( 250 mm × 4.6 mm × 5 μm) column, with acetonitrile ∶ water (4∶1) as mobile phase A and 30 mmol/L sodium acetate ∶ acetonitrile (355∶15, acetic acid adjusting pH value to be 6.5) as mobile phase B by gradient elution. The detection wave length was set at 254 nm. The flow rate was 1.0 mL/min. The column temperature was 40℃, and the injection volume was 5μL. Results Sixteen common peaks were shown in the fingerprint of 15 batches of honey samples. The similarity for 15 batches of honey samples was in the range of 0.910 ~ 0.996 . Conclusion The fingerprint detection method is simple, practical, reproducible and specific, and can provide certain reference for quality control of honey.

OBJECTIVE: The aim of this study was to explore the changes of the extracellular matrix in nasal mucosa by a guinea pig model of prolonged allergic-induced rhinitis. METHOD: Thirty-two male Hartley guinea pigs were randomly divided into four groups: allergen challenged groups (Group 2 w, Group 6 w and Group 12 w) and a control group. Ovalbumin-sensitized guinea pigs were repeatedly challenged with allergen twice a week from 2 weeks to 12 weeks. Matched control groups were challenged with physiological saline. Nasal mucosa were obtained from the animals killed. Hematoxylin-Eosin, Masson's trichrome, and immunohistochemical staining against transforming growth factor-beta1 (TGF-beta1), Collagen III and Collagen I were performed to nasal mucosa. RESULT: (1) Pathological examination showed obvious infiltration of eosinophils and the enlarged thickness of epithelial layer of nasal mucosa in the experiment groups. (2) The area ratios of blue stained in the extracellular matrix of nasal mucosa were increased. The area ratios of blue stained were statistically different in Group 6 w and Group 12 w compared with the control group. (3) The increasing absorbance of TGF-beta1 were statistically different in the experiment groups with the control group. The absorbance of Collagen III and Collagen I showed a rising trend along prolonged allergen challenged in the experiment groups. CONCLUSION Prolonged allergen challenge and the inflammation of nasal mucosa, can lead to the increasing of the inflammation relevant factors and the deposit of collagen in the extracellular matrix of nasal mucosa.
Allergens ; immunology ; Animals ; Collagen Type I ; metabolism ; Collagen Type III ; metabolism ; Eosinophils ; immunology ; Extracellular Matrix ; immunology ; metabolism ; pathology ; Guinea Pigs ; Inflammation ; Male ; Nasal Mucosa ; immunology ; metabolism ; pathology ; Rhinitis, Allergic, Perennial ; immunology ; metabolism ; pathology ; Transforming Growth Factor beta1 ; metabolism

OBJECTIVE: The aim of this study was to use a guinea pig model of prolonged allergic-induced rhinitis to characterize the feature of nasal mucosa remodeling. METHOD: Forty-eight male Hartley guinea pigs were randomly divided into six groups: allergen challenged groups (Group OVA(2w) , Group OVA(6w) and Group OVA(12w)) and control groups respectively (Group Sal(2w), Group Sal(6w) and Group Sal(12w)). Each group had 8 guinea pigs. To develop a guinea pig model of nasal mucosa remodeling, ovalbumin-sensitized guinea pigs were repeatedly challenged with allergen twice a week from two weeks to 12 weeks. Matched control groups were challenged with physiological saline. Nasal lavage was performed 24 hours after the last intranasal challenge. Then nasal mucosa were obtained. HE, AB-PAS, MT, and immunohistochemical staining against transforming growth factor-beta1 (TGF-beta1) were performed. Eosinophil cationic protein (ECP) and OVA-special IgE (OVA-sIgE) were detected by ELISA in nasal lavage fluid. RESULT: (1) The levels of OVA-sIgE in nasal lavage fluid in Group OVA(6w) and Group OVA(12w) were significantly different from Group OVA(2w), while the levels of ECP had no significant difference among the experiment groups. The levels of OVA-sIgE and ECP in experiment groups were significantly different from control groups respectively (P < 0.01). (2) Grade 0 and Grade 1 of epithelial damage were significantly different in Group OVA(6w) and Group OVA(12w) when compared with from Group OVA(2w) (P < 0.01). At the same time, Grade 0 and Grade 1 of epithelial damage were statistically different in the experiment groups when compared with the respectively control groups (P < 0.05). (3) Goblet gland hyperplasia and collagen deposit within the extracellular matrix (ECM) were easily found in Group OVA(6w) and Group OVA(12w) compared with Group OVA(2w) (P < 0.01). The number of goblet gland and the ratio of collagen deposit were statistically more in Group OVA(6w) and Group OVA(12w) than in Group Sal(6w) and Group Sal(12w) (P < 0.05). That feature of the ratio of collagen deposit did not show in Group OVA(2w) versus Group Sal(2w). (4) Increased TGF-beta1 expressions were observed in Group OVA(6w) and Group OVA(12w) compared with Group OVA(2w) (P < 0.01). Those increasing expressions were also observed in experiment groups rather than in the respectively control groups (P < 0.01). CONCLUSION Epithelial damage, goblet cells hyperplasia and collagen deposition in ECM were observed as the features of remodeling in this guinea pig model of allergic rhinitis under prolonged allergen challenge. Epithelial damage, excessive expression of related cytokines and enhancement activity of enzymes were observed in early time after challenge of allergen. The features of goblet cells hyperplasia and collagen deposition in ECM were observed at a later stage.
Airway Remodeling ; Allergens ; immunology ; Animals ; Disease Models, Animal ; Guinea Pigs ; Male ; Nasal Mucosa ; immunology ; pathology ; Rhinitis, Allergic, Perennial ; immunology ; pathology

BACKGROUND:Chitosan is a kind of natural biomaterial and is characterized by great biocompatibility, progressive degeneration and absorption and excellent mechanical property; however, whether it may become an ideal cytoskeleton in the engineering of cartilage tissue or not should be researched further.OBJECTIVE: To observe the hydrophilicity and adsorptivity to human nasal septum chondrocytes and the effect of its function of a novel scaffold made by [poly (dl-lactide-co-glycolide)] (PLGA)/chitosan nonwoven cloth embedded with lecithin (LEC) and poly-L-lysine (PLYS).DESIGN: Blank control study.SETTING: Department of Otolaryngology-Head and Neck Surgery, Nanjing General Hospital of Nanjing Military Area Command of Chinese PLA.MATERIALS: The experiment was carried out in the Department of Otolaryngology-Head and Neck Surgery, Nanjing General Hospital of Nanjing Military Area Command of Chinese PLA from October 2005 to June 2006. Chitosan nonwoven cloth was provided by Hainan Xinlong Company. The mainly technical parameters were detailed as the follows: degree of deacetylation ≥ 90% and relative molecular weight 2-5 × 105. PLGA/chitosan nonwoven cloth scaffold was made in Department of Otolaryngology-Hean and Neck Surgery, Nanjing General Hospital of Nanjing Military Area Command of Chinese PLA and High Polymer Institute of Sun Yat-sen University. The mainly technical parameters were detailed as the follows: mole ratio between monome lactide and glycolide 75:25, porosity 82%-86%, pore diamater 100-300 μm, shear strength 48 MPa, depth 1.5 mm and completely degenerated duration 14-18 weeks. Human nasal septum chondrocytes were the septal cartilage of nose which was derived from operated patients with deflection of nasal septum under sterile condition.METHODS: PLGA/chitosan nonwoven cloth was sheared into pieces with the size of 1.5 cm × 1.5 cm, dipped in 0.01 volume fraction of LEC anhydrous alcohol and 1 g/L PLYS for 6 hours, dealt with ultraviolet radiation after dehydration for 1hour, dipped in 0.75 volume fraction of ethanol for 24 hours, washed with Hanks solution, and incubated for 24 hours. After operations mentioned above, two kinds of novel scaffolds containing various components were obtained, and they were simple scaffold and scaffold embedded with LEC and PLYS. Cells derived from the third generation of human nasal septum chondrocytes were used to make suspension. In addition, cell suspension was grown on those two scaffolds to determine the degree of hydrophilicity through observing diffused degree of cell suspension. Whether cell suspension of human nasal septum chondrocytes was wafted on scaffolds or not were observed under phase contrast microscope so as to determine adsorptivity between cells and scaffolds; meanwhile, growth of cells and production of matrix were also observed.MAIN OUTCOME MEASURES: ① Hydrophilicity of those two kinds of scaffolds (diffused degree of cell suspension of human nasal septum chondrocytes on scaffold) and adsorptivity to cells (whether cell suspension of human nasal septum chondrocytes was wafted on scaffolds or not); ② growth of cells and production of matrix.RESULTS: ① When simple scaffold was put in cell suspension of human nasal septum chondrocytes, cell suspension showing like balls attached to the surface of scaffold, and then, scattered into space of scaffold gradually. During the period of culture, phase contrast microscope indicated that masses of cells attached to the surface of fiber of scaffold.When the petri dish was shaken, cell groups drifted irregularly. The adherent rate was (21±3.7)%. With the cultured time passing by, matrix was not produced. ② When scaffold embedded with LEC and PLYS was put in cell suspension of human nasal septum chondrocytes, cell suspension scattered into space of scaffold rapidly. The adherent rate was (89±5.6)%, which was higher than that of single scaffold group. This suggested that scaffold showed a strong hydrophilicity.Phase contrast microscope indicated that chondrocytes as the form of monome or community were distributed between scaffold and fiber averagely and attached to the surface of fiber of scaffold. When the petri dish was shaken, cell groups did not drift irregularly, and only a few of cells were scarred at the bottom of petri dish. This suggested that scaffold had a strong adsorptivity to cells. One week after culture, matrix showing like cobweb was produced among fibers of scaffold.With the cultured time passing by, matrix was produced abundantly.CONCLUSION: The novel scaffold of PLGA/chitosan nonwoven cloth embedded with LEC and PLYS is characterized by an excellent hydrophilicity and adsorptivity to human nasal septum chondrocytes; meanwhile, it also can secrete matrix.

Objective To investigate correlation between Ile50Val polymorphism of IL-4 receptor gene and asthma and the impact on plasm total IgE,sIL-4R level.Methods the DNA is isolated from 2ml venous blood from 100 nomal subject and 100 asthma subject.The genetic polymorphism was identified by Allele-specific PCR techniquers,plasm total IgE,sIL-4R level was determined by ELISA.Results The frequency of genetype is significantly different be- tween control group and asthma group x~2=18.94,P

OBJECTIVE To observe the ultra-structure and mucociliary transport speed of maxillary sinus mucosa of experimental rhinogenic acute sinusitis in rabbits.METHODS Fifteen healthy adult male New Zealand white rabbits were divided into experimental group(10 rabbits) and blank control group(5 rabbits) randomly.For the experimental group,a piece of polyvinyl acetal absorbent sponge(Merocel?) in size of 3 mm?5 mm?25 mm was inserted into the right-side nasal cavity of each rabbit.The sponges were soaked with 1ml type Ⅲ streptococcus pneumoniae solution.Two weeks after insertion,the mucociliary transport speed was measured by India ink solution method.All rabbits were sacrificed to obtain the right-side maxillary sinus mucosa for ultrastructure observation through transmission electron microscope.RESULTS The mucociliary transport speed of experimental group was much lower than that of blank control group,and the difference was statistically significant(P

Objective: Sampledrawing is an important procedure in the study of allergic airway inflammation.The authors investigate the methods of drawing samples from the animals with allergic airway inflammation.Methods: We included in this study 20 Guinea pigs,10 rats and 20 mice,which underwent trachea incision,followed by bronchoalveolar and rhinal lavage and collection of the lavage fluids.Then we collected blood samples via the heart from the guinea pigs and rats and via both the heart and the eyes from the mice,and obtained the tissues of the nasal cavities and lungs by different methods.Results: All the samples were satisfactorily obtained from the animals,and 80% of the bronchoalveolar and rhinal lavage fluids were collected.Conclusion: Different methods should be adopted to suit different sampledrawing from the animal models of allergic airway inflammation.

To explore the effects of the psychological behavior training on the recruits' physical performance. A total of 380 recruits selected from the Department of General Staff and missile troops were divided into experimental group and control group. The recruits of the experimental group undertook psychological behavior training before athletic exercises, including 3 000-meter running, grenade throwing, and push-up. The scores of these athletic exercises were compared between recruits of two groups. The results showed that the performance was improved in all the recruits, but the recruits of the experimental group made even greater improvement. The differences were statistically significant (P