Objective To construct an eukaryotic expression vector for TAP genes fused with enhanced green fluorescent protein(EGFP)gene,and to analyze the expression and subcellular localization of the fusion protein in A375 human malignant melanoma cells transfected with the eukaryotic expression vector.Methods A375 cells were cotransfected with the combination of plasmid(P)TAP1-EGFP or pTAP2-EGFP and pDsRed2-endoplasmic reticulum(ER),or with pEGFP-TAP1 and-TAP2,or monotransfected with pTAP1-EGFP or pTAP2-EGFP alone.The monoclonal A375 cells stably expressing TAP were obtained by G418 selection.Then.the distribution and expression of fusion proteins were assessed in A375 cells by using fluorescence microscopy and Western blot,respectively.Flow cytometry was used to measure the expression of HLA class Ⅰ on A375 cells.Results Transfection of A375 cells with pTAP1-EGFP or pTAP1-EGFP and pTAP2-EGFP significantly increased the expression of TAP 1 and TAP 2 in as well as HLA class Ⅰ antigen on A375 cells.The green fluorescence of TAP1-EGFP and TAP2-EGFP overlapped with the red fluorescence of ER marker in cotransfected cells.indicating that TAP was localized subcellularly on the ER.Conclusions The expression vector for TAP-EGFP fusion gene has been constructed cuccessfully and expressed in A375 cells,and the expressed fusion protein is subcelluiarly localized to ER.This study will provide a basis for the research into subsequent immune response following induction of TAP expression.

Objective To investigate the effects of social support intervention on anxiety and depression of pulmonary hypertension(PH)patients,then provide a scientific basis for nursing of patients with pulmonary hypertension.Methods The general condition of PH questionnaire,self-rating anxiety scale (SAS),self-rating depression scale(SDS),social support rating scale(SSRS)were distributed to 131 patients with PH.Then make statistical analysis of patients'anxiety,depression and social support conditions.Results The score of anxiety and depression psychological conditions of patients with pulmonary hypertension was significantly higher than normal population,the difference wag statistically significant.Among 131 patients,16 patients with anxiety,accounting for 12.21% ;21 cases of patients with depression,accounting for 16.03% .28 patients at a high level of social support,92 patients at a medium level of social support,11 patients at a low level of social support,a total of 91.60% of the patients in the middle and higher levels of social support.Anxiety and depression scores had significant negative correlation with social support,objective support points and subjective support points.The anxiety,depression difference among different types of pulmonary hypertension was statistically significant.The difference of anxiety and depression scores between patients with idiopathic pulmonary arterial hypertension and patients with pulmonary hyper tension caused by congenital heart disease were significant.The depression scores between pulmonary hy pertension caused by pulmonary veno-occlusive disease and congenital heart disease were significantly different.Conclusions When nurses care pulmonary hypertension patients.those with different types of PH should be given targeted social support.Attention should be paid to transfer the source of social support to help them adopt a positive attitude to face the diseabe,then improve the treatment and care compliance of patients.

Objective To study the changes of serum interleukin-8 (IL-8) and interleukin-10 (IL-10) in preterm infants delivered from mothers with chorioamnionitis (CA) and the possible effects on bronchopuhmonary dysplasia (BPD) and brain injury of preterm infants.Method From October 2014 to October 2015,clinical data from mothers without clinical manifestations of CA giving birth to a preterm baby (gestational age:26 to 33 weeks) were collected in Changzhou Maternal and Child Health Hospital and Suzhou Municipal Hospital.The infants were assigned to CA group and non-CA group according to their mother's placenta pathology.The levels of serum IL-8 and IL-10 were measured using enzyme-linked immunosorbent assay (ELISA) on day 1 and day 7,while the levels of WBC,CRP and PCT were measured at birth.The incidences of BPD and brain injury were also reviewed.Result A total of 67 preterm infants were included,with 51 in the CA group and 16 non-CA group.The levels of IL-8 were significantly higher in the CA group than the non-CA group on both day 1 and day 7 [(21.6 ±9.7) ng/L vs.(7.3 ±2.3) ng/L,(26.6 ± 12.9) ng/L vs.(7.3 ± 2.1) ng/L].The IL-10 levels were significantly lower on day 1 [(1.92±0.17) ng/Lvs.(2.04±0.18) ng/L] and higher on day 7 [(2.42±0.60) ng/L vs.(2.03 ±0.18) ng/L] in the CA group.Significant differences existed on the incidences of BPD (54.9％ vs.25.0％) and brain injury (74.5％ vs.43.8％) between the two groups (P ＜ 0.05).The levels of CRP and PCT were higher in CA group (P ＜ 0.05) and the WBC was similar between the two groups (P ＞ 0.05).In CA group,on both day 1 and day 7,infants with BPD had similar IL-8 and IL-10 levels comparing with infants without BPD(P ＞ 0.05),also were infants with brain injury comparing with infants without brain injury.Conclusion Chorioamnionitis in pregnant women may affect serum cytokines levels in premature infants and lead to high incidences of BPD and brain injury.

We discussed the influence of liquid nitrogen cryopreservation to survive capability of Babesia microti standard strain.The whole blood of mice infected with Babesia microti was put in liquid nitrogen to cryopreservation for 1 month,3 months,6 months,9 months,the whole blood was get out respectively and recovery at room temperature,and infected 3 mice respectively,100 μL/ mouse (the first generation after redissolution,the experiment group).In the same time,3 mice were also infected with Babesia microti as the animal conservation control group.When the infection rate was at a high level,the whole blood of the experiment group mice were injected into 3 normal BALB/c mice (the second generation after redissolution),to observe the changes of the Babesia microti form and proliferation situation,and also to observe the infection rate of the first and the second generation after redissolution in different conserving time.Compared with Babesia microti of animal subcultivation,the form of Babesia microti of liquid nitrogen cryopreservation changed a little.Small trophozoites,annular trophozoites,schizont and immature and mature merozoite and other form can also be seen.Compared with Babesia microti of animal subcultivation,the first time to see the worms and the time attaining to the high infection level were 1 to 2 days later,but for the second generation after redissolution,it is the same.There was no significant difference in different conserving time of 1,3,6,9 months.The influence of liquid nitrogen cryopreservation to survive capability and worm form of Babesia microti is a little,so liquid nitrogen cryopreservation can be a better way to conserving Babesia microti.

Objective To evaluate the sensitivity of sleep-deprivation electroencephalography(EEG) examination in diagnosis of partial seizures of children.Methods One hundred and six normal children (male 57,female 49, 3-12 years old)and 102 children suspected of epilepsy but with normal in standard EEG examination were selected at random(male 63,female 39,3-12 years old). Sleep-deprivation EEG was performed individually.Periods of sleep-deprivation were 18-20 hours for children below 7 years old and 24 hours for the older ones.Results The EEG showed slowing of background electric activity. In addition, 57 cases showed spindle/sharp-slow wave complex.The rate of EEG abnormality was 55.88%.The rates of EEG abnormality for partial seizures and generalized seizure were 76.32% and 43.8%,respectively(?~2=8.98 P

Objective To compare the sensitivity of sleep-deprivation electroencephalography(EEG)examination on children suffered with nonepileptic seizures in normal and suspected epileptic in children.Methods Thirty -eight children with nonepileptic seizures (male 13,female 25,age 4-12 years, mean 8.1 years) had induced a person to make a confession. One hundred and six normal children (male 57, female 49, age 3-12 years,mean 7.6 years) and 102 children(male 63,female 39,age 3-12 years,mean 7.3 years)suspected of epilepsy clinically but with normal standard EEG examination were selected at random as controls.Sleep-deprivation EEG was performed individually.Results The sleep deprivation EEG showed slow background electric activity in all 3 groups. In addition, the rates of EEG abnorma lity for nonepileptic seizures group were zero,no statistics were computed because F was a constant. Fifty-seven cases showed spindle/sharp-slow wave complex in suspected epileptic children.The rate of EEG abnormality was 55.9%.The rate of EEG abnormality for normal control group was 1.9 %.There were significant difference among 3 groups (?2=97.3 P

Adolescent ; Aortic Valve ; abnormalities ; Aortic Valve Insufficiency ; complications ; pathology ; surgery ; Aortic Valve Stenosis ; complications ; pathology ; surgery ; Child ; Endocarditis ; complications ; pathology ; surgery ; Female ; Heart Defects, Congenital ; complications ; pathology ; surgery ; Heart Valve Prosthesis Implantation ; Humans ; Male ; Rheumatic Heart Disease ; complications ; pathology ; surgery

Kawasaki disease is far more frequent in children than in adults. The pathogenesis of Kawasaki disease is unknown, but it involves changes to the coronary artery and other diverse clinical manifestations. There are currently no specific laboratory diagnostic indexes, and especially since the disease is rare in adults, so it is extremely easy to misdiagnose or to overlook entirely. Our retrospective analysis of an diagnosis of and treatment for Kawasaki disease in an adult provides a guide to clinical doctors in terms of understanding Kawasaki disease, early diagnosis of it, and improved prognosis.
Adult ; Aspirin ; therapeutic use ; Humans ; Male ; Mucocutaneous Lymph Node Syndrome ; diagnosis ; therapy ; gamma-Globulins ; therapeutic use

Objective To explore protective effect of Heme oxygenase-1(HO-1) on irradiation-induced endothelial cell apoptosis.Methods Human endothelial cell line EA.hy926 were administered with or without HO-1 activator Copp and/or HO-1 inhibitor Znpp,respectively.Then,cells were treated with or without 8 Gy radiation.The HO-1 protein expression of cells were assessed with Western blotting and apoptosis of cells treated with irradiation were evaluated with flow cytometry.Moreover,cytochrome C releasing into cytosol were also determined by Western blotting. Results In PBS+R group,HO-1 protein expression of EA.hy926 was low posterior to irradiation.When cells were preconditioned with Copp and/or Znpp,then recieved with 8Gy irradiation,the HO-1 protein expression of EA.hy926 increased significantly in comparision with the PBS+R group(P

To study the pharmacokinetic process of Danshensu in cerebal ischemia injury model rats and the correlation with its anti-cerebral ischemia effect. In this study, the middle cerebral artery occlusion (MCAO) model was established, in which all of the rats were intravenously injected of Danshensu at a single dose of 40 mg x kg(-1). The HPLC-DAD method was applied to determine the plasma concentration of Danshensu at different time points and draw the drug-time curve. Meanwhile, the superoxide dismutase (SOD) and the lactate dehydrogenase (LDH) activity were determined to draw the time-effect curve. The DAS 3.2. 6 software was used to process the data, analyze their correlation, compare the pharmacokinetic difference between model and normal rats after the administration of the same doses of Danshensu and the changes in pharmacodynamic indicators of model rats after the administration, and evaluate the effect of Danshensu in treating the cerebral ischemia disease. According to the results, the pharmacokinetic processes of Danshensu in the cerebral ischemia-reperfusion and normal rats were consistent to the two-compartment model. The main pharmacokinetic parameters were: t1/2alpha were (0.267 +/- 0.026), (0.148 +/- 0.020) h;t1/2beta were (1.226 +/- 0.032), (1.182 +/- 0.082) h; AUC0-infinity were (42.168 +/- 4.007), (26.881 +/- 1.625) mg x L(-1) x h. After the cerebral ischemia-reperfusion, the activity of SOD decreased and the activity of LDH increased. Danshensu could inhibit the decrease in the SOD activity and the increase in the LDH activity within a certain period of time. This indicated that Danshensu could stay longer in cerebral ischemia-reperfusion rats than in normal rats and eliminated more slowly, which reflected the rationality of Danshensu in the clinical treatment of cerebral ischemia diseases. Danshensu's effect against the cerebral ischemic injury may be related with its level in vivo. Its plasma concentration is positively related to the SOD activity and negatively related to the LDH activity.
Animals ; Brain Ischemia ; drug therapy ; Drugs, Chinese Herbal ; pharmacokinetics ; pharmacology ; therapeutic use ; Male ; Rats ; Rats, Sprague-Dawley ; Salvia miltiorrhiza ; chemistry