Animals ; Cytokines ; physiology ; Humans ; Immune System ; drug effects ; physiology ; Melatonin ; pharmacology ; Opioid Peptides ; physiology ; Receptors, Melatonin ; physiology ; Stress, Physiological ; immunology ; T-Lymphocytes ; drug effects ; physiology

Objective To investigate the expression and significance of Maspin and IKKα in nasosinusoidal mucosa of rats with fungal rhinosinusitis (FRS).Methods A total of 40 SD rats were used to establish the FRS model, and randomly divided into nasal obstruction group, FRS group, immunosuppressive group and invasive FRS group, 10 rats in each group.Another 10 normal rats were used as control group.Mice in the control group were fed with normal diet.In the nasal obstruction group, the mice had only hemostatic cotton stuffed in the nasal cavity and injection of 0.9% NaCl in the abdominal and nasal cavities.In the FRS group, the mice were injected Aspergillus fumigatus spore suspension into the nasal cavity and 0.9% NaCl i.p.The mice of the immunosuppressive group were given cyclophosphamide i.p.and 0.9% NaCl injection into the nasal cavity.The invasive FRS group was injected with cyclophosphamide i.p.and Aspergillus fumigatus spore suspension into the nasal cavity.The serum levels of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were measured by enzyme-linked immunosorbent assay (ELISA).The expression of Maspin and IKKα in nasosinusoidal mucosa was detected by immunohistochemical staining.The expression of Maspin mRNA and IKKα mRNA in the nasosinusoidal mucosa was detected by fluorescence quantitative PCR.Results The serum levels of IL-6 and TNF-α in different groups were significantly different (P< 0.05).The level of IL-6 in the FRS group was (69.3 ± 10.9) ng/L, significantly higher than those in the control group, nasal obstruction group, immunosuppressive group and invasive FRS group [(45.2 ± 7.1)ng/L, (46.4 ± 6.7) ng/L, (21.3 ± 4.5) ng/L, and (20.9 ± 4.3 ng/L)] (P < 0.05).The level of TNF-α in the FRS group was (30.4 ± 4.8) ng/L, significantly higher than those in the control group, nasal obstruction group, immunosuppressive group and invasive FRS group [(14.8 ± 2.7) ng/L, (13.9 ± 1.4) ng/L, (7.9 ± 0.6) ng/L, and (7.8 ± 0.4 ng/L)] (P < 0.05).The levels of IL-6 and TNF-α in the control group were significantly higher than those in the immunosuppressive group and invasive FRS group (P< 0.05).There was no significant difference between the immunosuppressive group and the invasive group (P> 0.05).Theresult of immunohistochemical staining showed that the protein expression of Maspin in the FRS group and invasive FRS group was significantly lower than that in the control group, nasal obstruction group and immunosuppressive group, while the expression of IKKα protein was significantly higher than that of control group, nasal obstruction group and immunosuppressive group (P< 0.05).The protein expression of Maspin in the invasive FRS group was significantly lower than that in the FRS group, by contrast, the expression of IKKα protein was significantly higher (P< 0.05).The PCRresult revealed that the expression levels of Maspin and IKKα mRNA were (0.217 ± 0.013) and (0.193 ± 0.012), significantly lower than that in the control, obstruction and immunosuppressive groups [(0.309 ± 0.021), (0.302 ± 0.017), and (0.293 ± 0.02)] (P< 0.05), while the expressions level of IKKα mRNA were significantly higher [(0.319 ± 0.043), (0.384 ± 0.048) vs (0.169 ± 0.015), (0.171 ± 0.018), and (0.175 ± 0.019)] (P< 0.05).Conclusions Down-regulation of Maspin expression after IKKα activation is the main cause of the onset of FRS, which may also be one of the mechanisms of invasive FRS.

OBJECTIVE:To establish a method for content determination of phosphatidylcholine in eustachian tube lavage fluid of patients with secretory otitis media. METHODS:HPLC was used. The samples were pretreated by liquid-liquid extraction. It was performed on the column of Hypersil CN with mobile phase of acetoneitril-methanol-phosphoric acid (100∶10∶0.6,V/V/V)at the flow rate of 1.8 ml/min,the detection wavelength was 205 nm,temperature was 30 ℃ and volume was 20 μl. RESULTS:The lin-ear range of phosphatidylcholine was 11.99-119.9 μg/ml(r=0.999 6);RSDs of precision tests of intra-day and inter-day were no more than 15%;average recovery was 97.54%(RSD=9.36%,n=9);the average content of phosphatidylcholine in eustachian tube lavage fluid of patients was(24.43±3.61)μg/ml. CONCLUSIONS:The method is simple and accurate,and can be used for the content determination of phosphatidylcholine in eustachian tube lavage fluid of patients with secretory otitis media.

Dust ; Female ; Humans ; Male ; Occupational Exposure ; adverse effects ; Pneumoconiosis ; diagnostic imaging ; etiology ; Poaceae ; Radiography, Thoracic ; Silicates ; adverse effects

Objective To investigate the association between Human Leukocyte Antigen DR (HLADR) gene polymorphisms and total IgE levels in children with asthma.Methods This study involved 84 unrelated children with asthma and 168 healthy controls without asthma.All participants had their serum total IgE levels measured with UniCAP Pharmacia system,and skin-prick test with ten kinds of inhalant allergens were taken among them.HLA oligonucleotide array was used to determine twenty-one gene frequencies of HLADR.Results ( 1 ) The frequencies of HLA-DRB1 * 070X allele and HLA-DRB1 * 11XX allele among the asthmatic were significantly higher than those in the healthy controls (HLA-DRB1 * 070X allele:2.98％vs.0.30％,x2 =6.915,P ＜ 0.05 ; HLA-DRB1 * 11XX allele:13.69％ vs.5.95％,x2 =9.478,P ＜ 0.01 ),Odds ratios( OR)for the two groups were 10.57(95％ CI:1.215 -91.986)and 2.79(95％ CI:1.429 -5.449)respectively.HLA-DRB3( 52 ) * 010X allele were significantly decreased in asthmatics compared to healthy controls(13.99％,x2 =5.854,P ＜0.05),OR was 0.429(95％ CI:0.214 -0.862).(2) Significant correlation between HLA-DRB1 * 160XX,HLA-DRB1 * 3 (17)alleles and the level of total IgE were found in asthmatic children(P ＜0.05).OR were 0.145(95％ CI:0.027 -0.781 )for HLA-DRB1 * 160XX allele and 1.667(95％CI:1.367-2.033)for HLA-DRB1 * 3(17)allele.Conclusion HLA-DRB1 *070X allele and HLA-DRB1 * 11XX allele were implicated in susceptibility to asthma,HLA-DRB3 (52) * 010X allele might conferring protection effects against asthma.There were association between HLA-DRB1 * 160XX,HLA-DRB1 * 3 ( 17 ) alleles and the level of total IgE in asthmatic children.Protective effects of HLA-DRB1 * 160XX allele against high level IgE response was noted,while HLA-DRB1 * 3(17)allele might be associated with high level of IgE in patients with asthma.

In order to explore the pathogenesis of non-alcoholic fatty liver disease (NAFLD), and to find the best evidence for clinical practice, recent literature about the pathogenesis and treatment of NAFLD was analyzed, and it was found that the generation of reactive oxygen species (ROS) is the most important factor in development of NAFLD. Based on insulin resistance (IR), generation of ROS is a central link in the course of "two hits". Other factors, such as leptin resistance, caspase-3, Fas and its ligand, peripheral natural killer T cells, cyclooxygenase-2, metabolic nuclear receptors, hepatic deposition of iron, ferritin, haptoglobin, retinol binding protein 4, imbalance of intestinal flora, mitochondrial dysfunction and endoplasmic reticulum stress, also contribute to the progress of NAFLD. In the treatment of NAFLD, beside the conventionally used methods such as IR improvement, antioxidation and lipid metabolism improvement, other medicines such as nuclear metabolism ligands or activators, iron-chelating agents and syndrome differentiation treatment in traditional Chinese medicine also have good efficacy.

Objective To discuss the effect of family nursing intervention on the life quality and pulmary function of patients with chronic obstructive pulmonary disease (COPD). Methods We divided 72 patients with COPD into the test group and the control group with 36 cases in each group.The two groups received routine treatment and nursing but additional family intervention was given to the patients and fam-ily members in the test group.The life quality and pulmonary function after intervention in the two groups were appraised in the two groups. Results The evaluation of life quality and pulmonary function were alleviated after intervention compared with those before intervention (P ＜ 0.05).But the above items in the control group were not significantly improved (P ＞ 0.05). Conclusions Effective family intervention could improve the life quality and pulmonary function of patients with COPD.

Objective To investigate the influence of knocking down Ezrin expression in combination with HSP70-induced immune killing on the apoptosis and proliferation of mouse osteosarcoma cells.Methods Human osteosarcoma cell line MG63 was cultured,the HSP70 and Ezrin-shRNA DNA fragments were cloned into the expression vector pGFP-V-RS containing CMV and pU6 promoters,and constructed the expression vector pGFP-V-RS-shRNA and pGFP-V-RS-shRNA-HSP70.The vectors were transfected into MG63 cell line,respectively.The status of transfected MG63 cells was observed by fluorescent microscope.The expressions of Ezrin and HSP70 were examined by real time RT-PCR and Western blot.Flow cytometry 、MTS test was used to detect the changes of cell apoptosis and proliferation,and changes of the expression of apoptosis and cell cycle related proteins was detected by Western blot.The specific cytotoxic T lymphocytes (CTLs) were induced by HSP70,and its killing effect on target MG63 tumor cells was analyzed by MTT assay.Results The specific vector simultaneously knocking down Ezrin and overexpressing HSP70 was constructed for the first time in China and confirmed by fluorescence microscope and gene/protein expression analysis.Compared to Ezrin knock-down alone,simultaneous HSP70 overexpression partially recovered the promoted cellular apoptosis and proliferation suppression by Ezrin knock-down,however,when compared to the normal control,the apoptosis rate of LM 8 cells was still significantly increased from 11.01％±0.22％ to 24.28％±0.50％,while the proliferation rate decreased from 395.14％±2.24％ to 310％± 2.83％.In addition,Western detected that Ezrin-shRNA could promote the expression of Bax.However,the expression of Ezrin-shRNA could reduce the Bcl-2 and Cyclin D1.Ezrin-shRNA/HSP70 also has the same effect.MTT assays revealed that the CTL cytotoxic effect on target MG63 tumor cells at all concentrations were significantly higher in CT+IL-2+HSP70 group compared with that in CT+ IL-2 group,with a cytotoxicity as high as 56.33％± 1.95％.Conclusion Simultaneous knocking down Ezrin and overexpressing HSP70 promotes the apoptosis and inhibits the proliferation of osteosarcoma cell.And the HSP70 can induce CTL which enhances the killing effect on tumor cell.And the HSP70 can induce CTL which enhance the killing effect on tumor cell.

Objective:To observe the pulmonary apoptosis resulted from ischemia reperfusion lung injury in rats. Methods: Forty male SD rats, weighing 300 350 g,were randomly divided into 8 groups: 6 ischemia reperfusion group,simple ischemia group and control group( n =5).An in situ ischemia reperfusion lung injury rat model was established. Apoptotic cells were detected by the terminal deoxynucleotidyl transferase mediate dUTP nick end labelling (TUNEL) technique and the apoptotic DNA fragments by pulmonary DNA electrophoresis. Electron microscopy was performed to verify the morphologic changes of apoptosis. Results: Obvious apoptosis of pneumocytes occurred early after(30 min) ischemia reperfusion lung injury, with the peak on 2 h after reperfusion, and there were no significant differences 12 h after reperfusion compared with control group. There was a slight but no statistically significant elevation of apoptotic pneumocytes number in simple ischemia group compared with controls. Conclusion: The result indicates that apoptosis may contribute significantly to ischemia reperfusion lung injury.

Objective To investigate the effects of neferine on the proliferation and the P-glycoprotein(P-gp) expression of refractory/relapsed acute leukemic cells and to provide experimental evidence for further clinical use. Methods MTT and immunocytochemistry SABC methods were used respectively to observe the alteration of the proliferation and the expression of P-gp in refractory/relapsed acute leukemic cells after treating with neferine. Results The inhibition ratio on acute leukemic cells of neferine adding adriamycin(ADM) group was significantly higher than that of ADM group (P0.05). Conclusion [WTBZ]Neferine can inhibit the proliferation of refractory/relapsed acute leukemic cells, and reduce the P-gp expression of refractory/relapsed acute leukemic cells and consequently reverse multidrug resistance(MDR).