BACKGROUND:At present,syndromes happening frequency of diabetes mellitus can be controlled through regulating glucose concentration in blood.Therefore,a fast,steady,accurate glucose biosensor has been an intense area of focus for investigators.OBJECTIVE:To observe the effect of chitosan-gold nanoparticles hybrid film on electrochemical glucose biosensor.DESIGN,TIME AND SETTING:The observation experiment was performed at the Department of Medicine Laboratory,General Hospital,Guangzhou Military Command of Chinese PLA from February to April 2009.MATERIALS:Based on the immobilization of glucose oxidase (GOD) with cross-linking in the matrix of chitosan on an Au electrode by glutaraldehyde,a novel glucose biosensor was constructed with gold nanoparticles by electrodeposition.METHODS:The properties of CHIT/AuNPs hybrid film modified electrode were characterized by cyclic voltammetry (CV) in 20 mmol/L K3Fe (CN)6-0.1 mol/L KCI system by using the three-electrode system and the response to glucose was investigated by linear scan voltammetry (LSV).The optimal experiment conditions,stability and reproducibility about the glucose biosensor were also studied.MAIN OUTCOME MEASURES:The cyclic voltammetry properties of different modified electrode,as well as relation between glucose concentration and response current.RESULTS:Glucose concentration showed a linear correlation with response current in range of 0.001-6 mmol/L.The biosensor exhibited excellent performances of producibility,stability and the ignorance interferences of reducing substances,with the CONCLUSION:The glucose biosensor possesses excellent reproducibility and stability with a fast response time to glucose,which can be used in detection of glucose concentration in human serum.

Iodine is an essential micronutrient for pregnant women,which affects thyroid function of pregnant women and fetus,and thus influences the intellectual development of offspring.Severe iodine deficiency could lead to overt hypothyroidism and fetal cretinism.In the areas implemented the universal salt iodization (USI) program,mild to moderate iodine deficiency and iodine excess may coexist during pregnancy.Iodine deficiency could lead to hypothyroxinemia in pregnant women.Iodine more than adequate and iodine excess could increase the prevalence of subclinical hypothyroidism.The effect of iodine supplementation on thyroid function in pregnant women with mild to moderate iodine deficiency may result in different conclusion.Therefore,it is important to maintain optimal urinary iodine concentration to prevent thyroid dysfunction.

Objective　To study the effects of prolactin on interactions between Graves′ disease (GD) thyrocytes and autologous peripheral blood mononuclear cells (PBMC) in coculture. Methods　Primary thyrocytes and autologous PBMC were cocultured at various levels of ovine prolactin (oPRL) in vitro. The activation and proliferation of PBMC as well as HLA-DR and CD40 expressions on thyrocytes were determined by immunofluorescent staining and flow cytometry. Results　 With PBMC in GD cocultures, oPRL caused significant increase in the percentage of CD4+CD25+ cells at 200 μg/L ［(13.08±2.54)%, P＜0.01］ and in the proliferative index at 200 μg/L ［(17.82±3.02)%, P＜0.01］ and 1 000 μg/L ［(16.57±2.56)%, P＜0.05］, but not at 12.5 μg/L ［(10.43±2.38)% and (14.08±2.67)%, respectively］ or 50 μg/L ［(9.94±2.54)% and (14.56±2.80)%, respectively)］. The values at 200 or 1 000 μg/L as above were also significantly different from those at 12.5 or 50 μg/L. The percentage of CD40+ cells ［(48.25± 6.63)%，(52.28±6.94)%］ and delta mean fluorescence intensity (dMF; 42.94±10.24，49.51±12.34) were significantly lower in thyrocytes in GD cocultures treated with 200 or 1 000 μg/L oPRL than those untreated ［(58.38±6.62)% and 67.30±20.20］ or treated with two other concentrations of oPRL. The percentage of HLA-DR+ cells ［(46.79±7.51)%, P＜0.01］ and dMF (21.02±5.43, P＜0.01) were significantly higher in the thyrocytes treated with 50 μg/L oPRL than those untreated ［(33.51±8.58)% and 13.91±3.88］ or treated with three other concentrations. There were no significant differences between the dose groups of oPRL but those mentioned above. Conclusions　Prolactin could affect interactions between GD PBMC and autologous thyrocytes and might eventually influence the shift of helper T cells. High prolactin levels might aggravate GD.

AIM To study protective effect of [D-Ala~2, D-Leu~5]-enkephalin (DADLE) against hydrogen peroxide (H_2O_2) induced myocardial cell injury and its possible mechanisms. METHODS Myocardial cells were isolated from neonatal rats and cultured for 48 h. Then the cells were randomly assigned into normal control, H_2O_2（200 μmol·L~(-1)）, H_2O_2+DADLE(1 μmol·L~(-1)), H_2O_2+DADLE +naltrindole(10 μmol·L~(-1)) and H_2O_2+DADLE +U0126(10 nmol·L~(-1)) groups and cultured for another 48 h.[~3H]TdR incorporation assay and flow cytometry were used to measure the cell proliferation and apoptosis rate. The lactate dehydrogenase (LDH) activities in culture supernatant measured by using LDH activity kit. The superoxide dismutase (SOD) activity and malondialdehyde (MDA) content in cells were measured with xanthine oxidase method and color reaction of thiobarbituric acid, respectively. The expressions of extracellular signal-regulated kinase (ERK) and phosphorylated-ERK (p-ERK) were observed with Western blot. RESULTS ① Compared with normal control group, the incorporation of [~3H]TdR in myocardial cells of H_2O_2 group was significantly lower, apoptosis rate was higher, LDH activity and MDA content in cells were higher, while SOD activity in cells was lower. In addition, the ratio of IA_( p-ERK) /IA_( ERK) was decreased. ② Compared with H_2O_2 group, the incorporation of ［3H］TdR in H_2O_2+DADLE group was significantly higher, apoptosis rate was lower, LDH activity and MDA content in cells decreased, while SOD activity increased significantly. The ratio of IA_( p-ERK) /IA_( ERK) was increased. ③ δ-Opioid receptor antagonist naltrindole and ERK antagonist U0126 inhibited this effect of DADLE on the above index changes induced by H_2O_2. CONCLUSION The δ-opioid receptor has protective effect against H_2O_2-induced myocardial cell injury, and its possible mechanism may be related to its promotion of antioxide capacity and ERK phosphorylation.

Objective To investigate the potential effects of Lactobacillus acidophilus strain L6 on prevention and treatment of Helicobacter pylori (H. pylori) infection in animal models. Methods A total of 200 hundred C57BL/6 mice were used in the study. ① Sixty mice were divided into control group (infected with H. pylori) and prevention group (previously treated with L6 and followed by infection with H. pylori) with 30 each. The incidence of H. pylori infection was compared between two groups.② Sixty mice were divided into control group (infected with H. pylori) and treatment group (infected with H. pylori for 4 weeks and followed by treatment with L6) with 30 each. Thechanges of H. pylori infection was compared between tow groups. ③ Eighty mice previously infected with H. pylori were orally administrated with L6 in the water supply over a period of 9 months. Of which, 40 H. pylori negative mice were either continuously or discontinuously treated with L6. The re-infection of H. pylori was compared between two groups. The H. pylori infection, gastric mucosal inflammatory and serum anti-H. pylori-lgG titer was measured by using urea breath test, histopathology and ELISA, respectively. Results The positive rate of H. pylori infection was 100% in control group and 20% in prevention group (pre-treated with L6 for 1 week). Whereas there was no H. pylori infection in the rest mice of prevention group pre-treated with L6 for 2 or 4 weeks. There was significant difference in H. pylori infection between two groups (P<0. 05).The serum anti-H. pylori IgG titer was lower in treatment group than in control group (P<0. 05).The reinfection of H. pylori in mice continuously treated with L6 was significantly lower than those discontiously treated with L6 (P<0. 05). Conclusions Preventively feeding L6 can significantly reduce the H. pylori infection in C57BL/6 mice, whereas the inhibition of H. pylori re-infection can also be achieved with long-term administration of L6.

Objective To analyze the synergistic effects between H.pyloriinfection and non-steroidal anti-inflammatory drugs use in peptic ulcer patients and upper gastrointestinal bleeding patients induced by peptic ulcer.Methods The peptic ulcer group consisted of 803 peptic ulcer patients,208 patients with gastrointestinal hemorrhage,and they were compared with 2 061 patients with non-peptic ulcer.Results H.pyloriinfection and NSAIDs use could increase the risk of peptic ulcer,and NSAIDs use in coordination with H.pyloriinfection in gastric ulcer morbility,but it's not significant coordination in the duodenal ulcer mortility.Pure NSAIDs use could increase the risk of bleeding gastric and duodenal ulcer,but pure H.pyloriinfection didn't increase the risk of bleeding peptic ulcer obviously,but the risk of bleeding peptic ulcer was not different in the patients of occasional,frequent and long-term NSAIDs use.Conclusion Detection of H.pyloriin the patients with long-term NSAIDs use is necessary,and eradication is needed in the patients infected with H.pylori.The NSAIDs use in peptic ulcer patients complicated with upper gastrointestinal bleeding should be payed attention and given timely treatment.

Objective To construct a kind of novel electrochemical glucose biosensor for clinical detection.Methods Multiwall carbon tube(MWCNTs),NanoAu and glucose oxidase(GOD) were immobilized on the Au electrode by monolayer self-assemble.Cyclic voltammetry and linear scan voltammetry were used to investigate the assembling process and the chemical characteristics of the glucose biosensor.Results The linearity of glucose in the range of 10～1 400 mg/L with the correlation coefficient of R2=0.990 9 was obtained by linear scan voltammetry and the detection limited reached 0.000 1 mg/L.The four criterion samples were measured with 3 different biosensors in same conditions and the RSD

To investigate the effect of arginine vasopression (AVP) on the secretion of vascular endothelial growth factor (VEGF) by vascular endothelial cell (VEC). VEC of human greater omentum artery were isolated and cultured. The concentrations of VEGF and activity of protein kinase C (PKC) in the culture medium were measured. Results indicated that after the cells were incubated with 1?10 -9～1?10 -7mol/L AVP for 6 hours, the concentration of VEGF and activity of PKC were markedly increased compared to control (P

OBJECTIVE To establish secretory otitis media(SOM) model induced by endotoxin and evaluate its reliability. METHODS Healthy 50 Sprague-Dawley rats of either sex( 250-300g)were randomly divided into experimental group(ET group) and control group(NS group),with 30 and 20 rats respectively. 35ul volume of 1mg/ml endotoxin from Pseudomonas aeruginosa and same amount of saline(NS) were transbullaly injected into the right tympanic bullas of the two groups. Six rats of ET group and four rats of NS group were killed postoperative on 6h,1d,3d,7d,14d respectively. The temporal bones were harvested for histological study. The left tympanic bullas served as control. Another four rats(each group 2 rats) were used for SEM study with the same method. One rat was killed and harvested for SEM observation of mucocilliary transporting system on the tympanic ostium of Eustachian tube. RESULTS ①LM(light microscope):The epithelial layer and subepithelial space(SES) were thicked mildly in ET group at 6 hours,and more heavily at 1 day with inflammatory cells infiltration (mostly polymorphonuclear leukocytes,PMNs). At 3 days,the same changes were observed most severely. By 14 days,the mucosal lining returned to normal. No obvious changes were observed in the NS group. ②SEM(scan electrical microscope):In ET group,the impairment of mucociliary transport system such as lodging,shedding of cilias were observed at 3 days and returned to normal at 14 days. No abnormal changes were observed in NS group. CONCLUSION Endotoxin can induce SOM with inflammatory changes peak at day 3 and return to normal by day 14.

Objective To observe the effect of ephedrine on the expression of eotaxin in human bronchial epithelial cells (16HBE) stimulated by tumor necrosis factor‐α(TNF‐α) and to explore the mechanism of Chinese medicine ephedra in treating asthma .Methods The in vitro cultured 16HBE were randomly divided into the control group ,TNF‐αstimulation group(TNF‐α20 ng/mL) and TNF‐αplus ephedrine group (TNF‐α20 ng/mL plus ephedrine 300 μg/mL) .Three complex holes in each group were set to culture for 18 h ,the eotaxin mRNA expression was measured by real time fluorescent quantified PCR and protein level was detected by immunocytochemical stain and Western blot .The eotaxin concentration in cells culture supernatant was quantified by ELISA .Results Compared with the the control group ,the expression level of eotaxin mRNA and protein ,and the concentration of eotaxin in cell culture supernatant in the TNF‐α stimulation group were increased obviously ,there being statisticaly significant difference between them(P<0 .01);however ,all above these parameters in the TNF‐αplus ephedrine group showed decreased obvi‐ously as compared with the TNF‐αgroup ,the difference between them was statistically significant (P<0 .01) .Conclusion Ephed‐rine can inhibit the expression and secretion of eotaxin in TNF‐α induced 16HBE inflammatory model ,which may be one of the mechanisms of Chinese medicine ephedra in treating asthma .