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 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.

Since the year 2006,I have posted evidences and papers on magazine,webs,and conference.(Ⅰ)Those posts and papers discuss relationship among origin of EEGs,information processing in brain,and blood circulation.(Ⅱ)Those posts and papers suggest that blood circulation plays the role of basic timer when brain processing information;Suggest one possible molecular mechanism of this'olood cycle timer'.In the molecular mechanism,H+,O2,working mechanism of microcirculation system,H+seats of NMDA receptor are key factors.(Ⅲ)Those posts and papers suggest that the sense of'time passes'is the sense of'press'that comes of blood flowing in brain.The sense of'a physical continuous process'is constituted by two senses:(i)Sense of'press'(ii)Sense come from'a sequence of active metadata'.This paper is a review of those evidences and papers.Section 1 gives a description of the model.Section 2～5 give the evidences supporting the model,and clarify some problems.Section 6～8 use the model to explain some phenomena of neuroscience(e.g.Origin of alpha wave,EEGs from kinds of sleep phases,etc.).

Objective: To study the mechanism of effect of total flavone of rhizoma drynariae in groups with different dosage on osteoblasts cultured in vitro. Methods: To culture osteoblasts with UMA-106-01 osteoblast strain and to observe the activity of ALP and the endosmosis of 3H-TdR. Results: The total flavone of rhizoma drynariae increased the activity of ALP in cells cultured with UMR-106-01 strain. The ALP activities correspondingly changed in the 24h, 48h and 72h, which related to dosage effect and time effect. Among them, the activity in the 48h is the most ideal one. And the amount of endosmosis of the total flavone of rhizoma drynariae to 3H-TdR increased more remarkably in the 48h than that in the 24h and it also related to the time effect. Conclusion: The total flavone of rhizoma drynariae can promote the differentiation and multiplication of osteoblasts.

Diabetic diarrhea is one of the chronic complications of diabetes, which is particularly closely related to the spleen and kidney. Using these two organs as the theoretical foundation, the root of diabetic diarrhea can be grasped. Pathogenic dampness is responsible throughout the whole process of diabetic diarrhea. Using syndrome differentiation, deficiency is as the basic pattern and excess is the syndrome for diabetic diarrhea; diagnosis and treatment combine symptoms and syndromes, which not only particularly pay attention to protecting the spleen and kidney, but also clearing pathogenic dampness. At the same time, flexible medication is applied according to dynamic symptoms, which can achieve good efficacy.

Objective To discuss the influence of gestational hypothyroxinemia to the pregnancy outcomes and fetus development,and find the evidence of hormone replacement therapy.Methods The clinical data of 1141 gravida admitted from Nov.2014 to Oct.2015 were retrospectively analyzed,including the data of systematic antenatal examination,all the data of pregnancy,the materials of delivery,the last ultrasound examination,production status and the thyroid stimulating hormone (TSH) of the newborn etc.,to find the difference of related index.Results Of the 1141 gravida with integral data,200 had past history of thyroid disease,189 showed below normal of free thyroxine (FT4) and 752 were normal ones.The 189 gravida with normal TSH but lower FT4 were divided into group A (0-5％ lower than the normal FT4 value,n=60),group B (5％-10％ lower than the normal FT4 value,n=40) and group C (10％ and above lower than the normal FT4 value,n=89).The ones with both normal TSH and FT4 value served as control group.Compared to the control group,the higher premature delivery rate,incidence of gestational diabetes mellitus and cesarean delivery rate (P＜0.05) were found in group C,and more gravida in group B had a history of hypertension and dyslipidemia during pregnancy (P＜0.05).The cesarean delivery rate of group B and C were higher than group A.Meanwhile,the rate of group B was higher than control group (P＞0.05).At delivery,the maternal weight,BMI,diastolic pressure,and head circumference of fetus in the last ultrasound examination were higher in group C than in control group (P＜0.01),but the gestational weeks of the newborn were shorter in group C (38.55 ± 1.86 weeks) than in control group (39.14 ± 1.57 weeks,P＜0.01).The 189 gravida with lower FT4 were divided into two groups according to the thyroid peroxidase antibody (TPOAb) level.The head circumference of fetus in the last ultrasound examination was higher in TPOAb(+) group than in TPOAb(-) group (45.99 ± 62.36cm vs.33.23 ± 2.08cm,P＜0.01).Conclusions The influence of gestational hypothyroxinemia to pregnancy outcomes and fetus development cannot be ignored,especially for the pregnant women with lower FT4 value (10％ and above lower than the normal) or with positive TPOAb.It is suggested to take the thyroid function test in the early stage of pregnancy for those pregnant women mentioned above.

Objective To investigate the expression and significance of Toll-like receptors-4 in peripheral blood and placental tissue in gestational diabetes mellitus (GDM) patients.Methods From February 2013 to February 2015.a total of 30 cases of gestational diabetes mellitus patients(GDM group) and 30 cases of normal pregnant people(health group)were selected as research objects.Peripheral blood and placental tissue of two groups were collected.The expression of TLR4 mRNA in peripheral blood mononuclear cells was detected by RT-PCR,the expression of TLR4 protein in placenta tissue was detected by immunohistochemistry.Results The expression of TLR4 mRNA in peripheral blood mononuclear cells of the observation group was significantly higher than that of the health group[(0.63±0.12) vs.(0.32±0.07)],the difference was statistically significant(t=12.223,P<0.05).The expression of TLR4 in villous trophoblast cells,decidual cells and amniotic epithelial cells in GDM group was significantly higher than that in the health group(Z=2.325,2.374,2.162,P<0.05).Conclusion The expression of TLR4 in peripheral blood mononuclear cells,villous trophoblast cells and decidual cells in gestational diabetes mellitus patients significantly enhanced,suggesting that TLR4 might be related to the e gestational diabetes mellitus.