Objective To investigate the expression of neuronal extracellular signal-regulated kinase 1/2 (ERK1/2) and its significance after cerebral isehemia reperfusion in diabetic rats. Methods Seventy-two healthy adult SD rats were randomly divided into sham-operation, normal glucose with cerebral ischemia and diabetes with cerebral ischemia groups. Each group was redivided into ischemia 15 minutes and reperfusion 1, 3 and 6 h subgroups according to the different time points of ischemia reperfusion (n = 6 in each subgroup). Streptozocin was used to induce diabetes, and a global cerebral ischemia model of diabetic rat was established by the bilateral vascular occlusion combining with bloodletting, TUNEL and immunohistochemistry were used to observe neuronal apoptosis and the expression of the phosphorylation of ERK1/2 in hippocampal CA4 region. Results The incidences of neuronal apoptosis in hippocampal CA4 region for ischemia 15 minutes and reperfusion 1, 3 and 6 h in the diabetes with cerebral ischemia group were significantly higher than those in the normal glucose with cerebral ischemia group (P < 0. 05); the expressions of the phosphorylation of ERK1/2 at all time points in the diabetes with cerebral ischemia group were higher, and reperfusion 1 and 3 h were significantly higher than those in the normal glucose with cerebral ischemia group (P < 0.01). Conclusions ERK1/2 might involved in the mechanism of neuronal injury after diabetes aggravating cerebral ischemia-reperfusion.

Objective To explore the expression of extracellular signal-regulated kinase 1/2 (ERK1/2) in the hippocampus CA4 region of diabetic rats after global cerebral ischemia-reperfusion injury. Methods Diabetic rat was established by intraperitoneal injection of streptozocin (STZ) and then global cerebral ischemia model was induced by bilateral clamping of the carotid arterial plus hypotension by withdrawing blood. Apoptosis of neuron and expression of the phosphorylation of ERK1/2 (P-ERK1/2)were observed in the hippocampus CA4 region of diabetes operation groups (DCI) and normoglycemia operation groups (NCI) by TUNEL,immunohistochemistry at 15 min after ischemia and at 1 h after reperfusion. Results Compare with the NCI,neuronal apoptosis of DCI was significantly higher at each time point of cerebral ischemia and reperfusion in the hippocampus CA4 (P

Objective To explore the influence of Akt inhibitor MK2206 in the proliferation and apoptosis of tongue squamous cell carcinoma TCA-8113 cells,and to clarify the possible mechanism.Methods The tongue squamous carcinoma TCA-8113 cells at the logarithmic phase were randomly divided into control group and 1,5,25,125, 250 nmol·L-1 MK2206 groups.The inhibitory rate of proliferation of TCA-8113 cells was detected with MTT method,and the apoptotic rate of TCA-8113 cells was determined with flow cytometry(FCM),and the expressions of caspase-9,Bad,GSK-3β,p-Akt and T-Akt proteins in the TCA-8113 cells were detected with Western blotting method.Results The IC50 of tongue squamous cell carcinoma TCA-8113 cells after treated with MK2206 for 12, 24,and 36 h were (112.54±1.67),(79.67±2.01),and (33.33±1.98)nmol·L-1 .The FCM results showed that the apoptotic rates of TCA-8113 cells after treated with 1,5,25,125,and 250 nmol·L-1 MK2206 for 12 h were (14.2±0.74)%,(19.3±0.45)%,(35.1±0.45)%,(39.6±0.48)% and (52.1±0.19)%;there were significant differences compared with control group(P<0.01).The Western blotting method results showed that the expressions of p-Akt, Bad and GSK-3βwere decreased with the increasing of dose and time of MK2206;compared with theβ-actin in control group,the bands got darken;the expression level of caspase-9 was increased, compared with theβ-actin in control group, the bands got darken;the T-Akt protein expression did not change significantly;compared with the β-actin in control group, the color of bands had no significant difference.Conclusion Akt inhibitor MK2206 can inhibit the proliferation of tongue squamous cell carcinoma TCA-8113 cells and induce apoptosis.

Objective To explore whether arsenic trioxide(As2O3)-induced apopotosis in drug-resistant leukemia K562/ADM cells may induce in through endoplasmic reticulum stress leukemia cell apopotosis.Methods The apoptosis of K562/ADM cells was identified by double staining of FITC-Annexin V and propidium iodide(PI),the ultrastructure of the cells,endoplasmic reticulum and mitochondria were observed by transmission electron microscopy.Flow cytometry(FCM) was employed to assess mitochondrial inner membrane potential(??m),intracellular calcium concentration,cytochrome c(Cyt c) release and caspase-3 activity.The expression of GRP78 protein was analyzed by Western blot.Results During the apoptotic process of K562/ADM cells induced with 2 ?mol/L and 5 ?mol/L As2O3,the endoplasmic reticulum exhibited obvious expansion and degranulation,and the mitochondria illustrated inner and outer membranes fusion,reduced and confused cristae,swelling and vacuolization.The mitochondrial ??m decreased,the intracellular calcium concentration and releasing of cytochrome c from mitochondria increased,and caspase-3 was activated.Western blot result indicated upregulation of GRP78 protein at endoplas-mic reticulum in apopototic K562/ADM cells.Conclusion As2O3 can initiate the endoplasmic reticulum stress in K562/ADM cells,and induces to apoptosis of the drug-resistant cell via endoplasmic reticulum-mitochondrial pathway.

Adult ; Cardiomyopathy, Hypertrophic ; pathology ; Female ; Heart Ventricles ; pathology ; Humans

This study was performed to prepare immobilized β-glucosidase and snailase, then optimize and compare the process conditions for conversion of icariin. Immobilized β-glucosidase and snailase were prepared using crosslink-embedding method. The best conditions of the preparation process were optimized by single factor analysis and the properties of immobilized β-glucosidase and snailase were investigated. The reaction conditions including temperature, pH, substrate ratio, substrate concentration, reaction time and reusing times of the conversion of icariin using immobilized β-glucosidase or snailase were optimized. Immobilized β-glucosidase and snailase exhibited better heat stabilities and could remain about 60% activity after storage at 4 degrees C for 4 weeks. The optimized conditions for the conversion of icariin were as follows, the temperature of 50 degrees C, pH of 5.0, enzyme and substrate ratio of 1 : 1, substrate concentration of 0.1 mg x mL(-1), reaction time of 6 h for β-glucosidase and 2 h for snailase, respectively. In 5 experiments, the average conversion ratio of immobilized β-glucosidase and snailase was 70.76% and 74.97%. The results suggest an effect of promoted stabilities, prolonged lifetimes in both β-glucosidase and snailase after immobilization. The immobilized β-glucosidase and snailase exhibited a higher conversion rate and reusability compared to the free β-glucosidase and snailase. Moreover, the conversion rate of immobilized snailase was higher than that of immobilized β-glucosidase. The process of icariin conversion using immobilized β-glucosidase and snailase was moderate and feasible, which suggests that immobilized enzymes may hold a promise for industrial usage.
Enzymes, Immobilized ; chemistry ; Flavonoids ; chemistry ; Hydrolysis ; Temperature ; beta-Glucosidase ; chemistry

Objective: To observe the clinical efficacy of combining auricular point sticking and a healthy diet to treat simple obesity in children aged 6-9 years old.Methods: A total of 190 eligible obese kids were divided into an observation group and a control group using the random number table method, with 95 cases in each group. The observation group was intervened by auricular point sticking plus guide on a healthy diet, while the control group was only provided with the guide on a healthy diet. The therapeutic efficacy was observed after intervention for three consecutive months, as well as the changes in body mass (BM), body mass index (BMI), waist circumference (WC), hip circumference (HC), and subcutaneous fat thickness. Results: After the 3-month intervention, the total effective rate was 91.6％ in the observation group, versus 74.7％ in the control group, and the between-group difference was statistically significant (P<0.01); in both groups, the BM, BMI, WC, HC, and subcutaneous fat thickness all decreased significantly (P<0.05), and were lower in the observation group than in the control group, showing statistical significance (P<0.05). Conclusion: Auricular point sticking plus a healthy diet is safe and effective in treating simple obesity in children, producing more significant efficacy than healthy diet intervention alone.

To assess the level of health skills relating to infectious diseases among residents in Beijing and develop more effective strategies for controlling infectious diseases.We conducted a multistage sampling,retrospective cross-sectional survey of residents aged 18 and above from 6 districts by selfadministered and anonymous questionnaires in January 2011.The questionnaires consisted of health skills relating to infectious diseases.A total of 13 042 respondents completed the survey.The proportions of people with the skills for acquiring knowledge about infectious diseases,being able to read medicinal package insert,being able to read brochures with regard to basic knowledge about infectious diseases, being able to read laboratory test report andproper use of thermometer were 57.7％,71.0％,70.2％,47.9％ and 76.8％ respectively.The levels of health skills were different among different populations (P ＜0.01).A low level of health skills was observed among the poorly-educated people and elders.Therefore measures such as health education and skills training about infectious diseases should be taken to improve the health skills.

Objective To explore the clinical value of renal dynamic imaging and urinary N-acetyl-?-D-glucosaminidase(NAG),apoptosis DNA fragment(ADF) in evaluating the damage degree of hydronephrotic kidneys(HnK) in children with hydronephrosis.Methods Level of glomerular filtration rate(GFR) was detected in 41 children with congenital hydronephrosis by renal dynamic imaging,and urine NAG,ADF in pelvis in HnK and healthy kidneys (as controls) were detected by enzyme-linked immuno-sorbent assay(ELISA).Patholo-gic changes of HnK in 41 children were graded intoⅠ~Ⅴ according to Elder standard. And GFR,urinary NAG and ADF of HnK were divi-ded into subgroup according to pathologic changes ,at the same time statistical analysis was performed within each groups. And the correlations of pathologic grades with GFR,urinary NAG and ADF of HnK were analyzed.Results 1.Kindneys GFR in healthy kidneys and Hnk were (174.33?20.43)?10-3 L/min,(143.86?17.51)?10-3 L/min respectinely,and there was significant difference between healthy kidneys and Hnk (P0.05).3.There was significant negative correlation between GFR levels of HnK and pathologic grades(r=-0.814 P0.05).Conclusions For hydronephrotic kidneys,urinary NAG can eva-luate impaired nephric tubule whereas renal dynamic imaging may evaluate the damage level of glomeruli;urine ADF may not indicate the damage level of diseased kidneys in children with congenital hydronephrosis.

Objective To explore the expression and role of LAT 1 in mouse uterus on early placenta formation on day 8 of pregnancy (D8).Methods One hundred and twenty 6-8-week old SPF female Kunming mice were used in this study.Immunohistochemistry was applied to determine the localization of LAT 1 protein in the mouse uterus on D8 of preg-nancy.The ectoplacental cones (EPCs) were dissected out from D8.5 uterus, and then cultured in vitro with different con-centrations of BCH (2-aminobicyclo-(2,2,1)-heptane-2-carboxylic acid, specific antagonist of LAT1) and L-leucine ( substrate of LAT1) to determine the role of LAT1 during the EPC attachment and outgrowth .Results LAT1 protein was highly expressed in secondary decidual zone and also positively expressed in the mouse uterus on D 8.As a specific antago-nist of LAT1, BCH significantly suppressed the ectoplacental cone outgrowth , whereas L-leucine showed no significant effect on it.Conclusions LAT1 is expressed in the mouse uterus during early placenta formation and promotes ectoplacen -tal cone outgrowth , suggesting that LAT1 may promote the trophoblast invasion into maternal decidual tissue , and partici-pates in the early formation of placenta .