Objective To evaluate the changes in the expression of DJ-1 protein during myocardial ischemia-reperfusion (I/R) in diabetic rats.Methods Fifty male Sprague-Dawley rats,weighing 220-280 g,were used in this study.Type 1 diabetes mellitus was induced by intraperitoneal streptozotocin 65 mg/kg and confirmed by fasting blood glucose ＞ 16.7 mmol/L.Forty animals with type 1 diabetes mellitus were randomly divided into 3 groups:diabetes group (group D,n =10),diabetic sham operation group (group DS,n =15) and diabetic I/R group (group DIR,n =15).Another 10 non-diabetic rats in which citrate buffer 6 ml/kg was injected intraperitoneally were served as control group (group C).Myocardial I/R was produced by occlusion of the anterior descending branch of left coronary artery for 30 min followed by 120 min reperfusion in group I/R.At 120 min of reperfusion,5 rats were sacrificed and myocardial specimens were c(on)tained for determination of infarct size in groups DS and DIR,and 10 rats were sacrificed and myocardial specimens were obtained for microscopic examination and for determination of cell apoptosis,malondialdehyde (MDA) content,superoxide dismutase (SOD) activity and expression of DJ-1 and phosphatase and tensin homologue (PTEN) protein.Apoptotic index (AI) was calculated.Linear correlation between the expression of DJ-1 protein and MDA content,SOD activity,AI and expression of PTEN protein was analyzed.Results Compared with group DS,the myocardial infract size was significantly increased in group DIR (P ＜ 0.05).Compared with group C,MDA content and AI were significantly increased,SOD activity was decreased,the expression of DJ-1 was down-regulated,and the expression of PTEN protein was up-regulated in groups D,DS and DIR (P ＜ 0.05).Compared with groups D and DS,MDA content and AI were significantly increased,SOD activity was decreased,the expression of DJ-1 was down-regulated,and the expression of PTEN protein was up-regulated in group DIR (P ＜ 0.05).There was no significant difference in the parameters mentioned above between groups D and DS (P ＞ 0.05).There was linear correlation between the expression of DJ-1 protein and MDA content,SOD activity,AI and expression of PTEN protein and the correlation coefficients (r) were-0.734,0.593,-0.818,and-0.812 in turn.Conclusion Down-regulation of DJ-1 protein expression is involved in myocardial I/R injury in diabetic rats via decreasing anti-oxidative stress responses and upregulating PTEN protein expression.

Objective To explore the effect of rhynchophylline and isorhynchophylline on headshakes behavior and levels of monoamine neurotransmitter in model rats with Tourette syndrome.Methods 40 Wistar rats were randomly divided into DOI-induced head-shakes rats (HSR group),haloperidol group,rhynchophylline group and isorhynchophylline group with 10 in each group.The inhibitory effects of rhynchophylline and isorhynchophylline were estimated by observing the HSR behavior.Dopamine (DA) and 5-hydroxytryptamine(5-HT) in the rat striatum were detected by the enzyme-linked immunosorbent assay (ELISA) method.The 5-HT2A receptor mRNA expression in prefrontal lobe cortex of the rats was measured by real-time PCR.Results Compared with HSR group,the head shakes of the rats in haloperidol group and isorhynchophylline group were significantly decreased(P＜0.01),and no change of head-shakes number was observed in rhynchophylline group (P＞0.05).There was no significant difference of head-shakes number between the haloperidol group and isorhynchophylline group(P＞0.05).Compared with HSR group,DA levels in the rat striatum were significantly decreased in isorhynchophylline group and haloperidol group((152.35± 5.80) μ~L vs (111.19±4.30) μg/L,(152.35±5.80) μg/L vs (126.42±3.17) μg/L,P＜0.01),while DA levels in the rat striatum in rhynchophylline group were not changed ((152.35±5.80) μg/L vs (142.71±5.51) μg/L,P＞0.05).There was no significant change of 5-HT2A receptor mRNA expression in rat prefrontal lobe cortex in every group(P＞0.05).Conclusion Isorhynchophylline may have an inhibitory effect on rats with DOI-induced HSR.Isorhynchophylline may decrease the DA levels in the rat stratum with DOI-induced HSR.Rhynchophylline has no significant inhibitory effect on head-shakes behavior and DA levels in the rat stratum with DOI-induced HSR.

Objective To analyze the genotype-phenotype correlations of Angelman syndrome ( AS ) , and to discuss the advantage of applying array-based single nucleotide polymorphisms comparative genomic hybridization ( SNP aCGH) in diagnosis of AS. Methods Examination of electroencephalogram( EEG) and intelligence quotient( IQ) evaluation were done for 11 cases diagnosed as AS clinically. Gesell scares were chosen as the evaluation criterion of IQ. The screening techniques was methylation polymerase chain reaction( MS-PCR) ,then SNP aCGH was used to make genetic diagnosis. Results (1)Eleven cases of AS were confirmed:1 case had UPD(uniparental disomy),10 cases were type of deletion, from which 6 cases were deletion (Ⅱ) , 4 cases were deletion (Ⅰ) . ( 2 ) The copy number variations were detected in the region of 15q11-q13,which contained genes like MKRN3,MAGEL2,NDN,SNRPN, SNURF,GABRB3,GABRA5,GABRG3,UBE3A,OCA2,ATP10A. To search online Mendelian inheritance in man,genes above were correlated with AS manifestation. (3)All cases of deletion were 3-5 standard deviation(SD) in weight and height to normal children at the same age and with the same sex,while UPD was below 1. 5 SD. Gesell scares showed that the deletion(Ⅰ) was the most serious in mental retardation,deletion(Ⅱ) was moderate,and the UPD was mild. Eight cases were hypopigmentation,and one was the UPD. EEG revealed that 1 case of deletion(Ⅰ) and the UPD were spike occasionally,another one deletion(Ⅰ) was limit EEG. The rest cases displayed slow and spike waves paroxysmal-ly,with amplitude of medium or high,2. 5-3. 0 Hz. Conclusions Not only can SNP aCGH make a diagnosis of AS but discriminate the types of genetic pathology. Since different type contributes to a diverse of clinical features and the rate of recurrence is also different,it is significant for family genetic consultation. Moreover,the technology is advantageous for the study on the pathogenesis and gene function.

Objective:To prepare Dermatophagoides farinae (Der f) crude protein to establish BALB/c bronchial asthma model , and to observe the morphology and degranulation of mast cells and detect related cytokines .Methods: Dermatophagoides farinae ( Der f) crude protein were prepared by trituration .30 BALB/c mice were randomly divided into 3 groups:PBS control group (A), asthma model group (B) and Der f crude protein treatment group (C).Group A were treated with PBS(100 μl) all the time, group B and group C were treated with 50 μg Der f crude protein mixed with 50μl alum adjuvant on day 0,day 7 and day 14.On day 28 group A and B were subcutaneous injected with PBS (100 μl) and group C were subcutaneous injected with Der f crude protein (350μg) in PBS(100 μl) at 1-day intervals.One week after the last treatment ,group A,B and C were intranasally challenged with 50 μg Der f crude protein daily for seven days .Twenty-four hours after the last challenge , airway hyper-responsiveness ( AHR) was assessed by using whole-body plethysmography .Two days post challenged , mice were sacrificed and bronchoalveolar lavage fluid ( BALF) was collected.Number of the total cells and eosinophil was determined .Level of IL-4,IL-10 and IFN-γcytokines in the BALF and the su-pernatant of splenocyte culture was assayed by ELISA .Level of Der f specific IgE and histamine in the sera was determined by ELISA . Airway inflammation was analyzed by HE staining .Observation of the morphology and degranulation of mast cells was analyzed by tolui -dine blue staining.Results:Compared with group B,AHR and the lung inflammation in group C were greatly reduced (P<0.01). Numbers of total cells and eosinophils in BALF of group C were significantly lower than that of group B ( P<0.01 ) .Compared with group B, the observation of degranulation of mast cells was insignificant in group C .Compared with group B(IgE:1.905), the level of specific IgE was significantly lower in groups C (IgE:1.278)(P<0.01).The level of IL-4 in BALF of group C was significantly lower than that of group B(P<0.01).Compared with group A and B, the level of IL-10 in BALF was significantly higher in group C (P<0.01) and the level of IFN-γin BALF of group C was significantly higher than that of group A and B (P<0.01).Compared with group B, the level of IL-4 in cultured splenocytes was significantly lower in group C (P<0.01), and the level of IL-10 and IFN-γin cultured splenocytes of group C was significantly higher than that of group B (P<0.01).Compared with group B, the level of histamine in BALF was slightly lower in groups C (P<0.05), and the level of histamine in sera was significantly lower in groups C (P<0.05). Conclusion:The degranulation of mast cells of murine bronchial asthma model was suppressed after the desensitization therapy .

Background and purpose:Visceral pleural invasion (VPI) and vessel invasion (VI) are poor prognostic factors in patients with non-small cell lung cancer (NSCLC). The primary initial recurrence site may be local recurrence in VPI and distant metastasis in VI. The purpose of this study was to validate the prognostic impact and effect of the initial recurrence site of VPI and VI on survival outcomes for NSCLC.Methods:Two hundred and ninety patients who were diagnosed as having NSCLC and underwent lobectomy between Jan. 2007 and Dec. 2013 were retrospectively analyzed. VPI was identiifed in 51 patients as VPI group, the other 239 patients without VPI as non-VPI group. VI was identiifed in 29 patients as VI group, the other 261 patients without VI as non-VI group. Clinical characteristics, overall survival (OS), disease-free survival (DFS) were compared.Results:There were statistically signiifcant differences between VPI group and non-VPI group in tumor size, lymph node metastasis, TNM stage and initial recurrence site (P<0.05). Furthermore, there were statistically signiifcant differences between VI group and non-VI group in lymph node metastasis and TNM stage (P<0.05). The 1-, 3- and 5-year OS rates in VPI group (88.2%, 56.7% and 52.7%) were lower than those in non-VPI group (95.8%, 83.7% and 74.0%,P<0.001). The 1-, 3- and 5-year OS rates in VI group (79.3%, 56.8% and 48.7%) were lower than those in non-VI group (96.1%, 81.3% and
72.3%,P=0.001). Cox regression showed TNM stage was a significant prognostic factor for DFS, whereas lymph node metastasis and VPI were signiifcant prognostic factors in patients with NSCLC.Conclusion:The primary initial recurrence site in VPI patients is local recurrence. Patients with VPI or VI may need more postoperative therapy because of their poor prognosis.

Objective By using array-based single nucleotide polymorphisms comparative genomic hybridization (SNP-aCGH) to detect and fine mapping copy number variations (CNVs) in children with unexplained mental retardation/brain development delay(MR/BD),then the CNVs were analyzed to determine diagnosis and offer genetic counseling,finally to discuss the application of SNP-aCGH in genetic diagnosis of MR/BD with unknown causes.Methods Ninety-two children with unexplained MR/BD were recruited.SNP-aCGH was used to get CNVs from the whole genome-wide,and the correlation of CNVs and phenotype was analyzed to definit disease genes or pathogenic fragment.Statistics was performed to analyze the common phenotype between positive cases (case with CNVs) and negative cases.Results (1) The CNVs were detected in 10 cases with a detection rate of 10.86％,from which 8 cases showed subtelomeric aberration,5 cases without subtelomeric aberration,and the rate was 8.70％,5.40％,respectively.The CNVs related to MR/BD involved 10 different subtelomeric regions (9p,21q,3p,2p,15q,4p,12p,22q,16p,17p),and 7 different regions without subtelomeric (1p,4q,2p,14q,15q,12q,22q).The deletions involved 11 zones (size:1.05-8.80 Mb),and duplications referred to 8 zones (size:1.33-31.25 Mb).(2) One case was diagnosed as 9p duplication syndrome,for candidate genes:DOCK8,VLDLR.A case was detected with a gene fracture (CRBN).One case was diagnosed as Coffin-Sirrs syndrome combined with a deletion of 15q26.3-qter,for candidate genes:SOX11 and LINS1,respectively.One case referred to 12p13.3 deletion syndrome,for candidate genes:ELKS,ERC1.One case referred to 22q13.2-qter deletion,for candidate genes:SHANK3.Two cases were diagnosed as ATR-16 syndrome with 17p13.3 deletion syndrome,their candidate genes:HBA1,HBA2,SOX8 for the former,YWHAE,LIS1 for the latter.(3)There were statistically significant differences in comparison of positive cases to the negative ones for growth delay,internal organs deformity,low birth weight infant(LBW) and premature infant (all P ＜0.05).Conclusions (1) Besides MR/BD in different degrees in all the positive cases,they also showed growth delay,a portion of them with internal organs deformity,low birth weight infant and premature infant.(2) Subtelomeric aberrations are related to MR/BD,while the submicroscopic rearrangement in regions without subtelomeric is suspiciously pathogenic,and need to be further studied.(3) SNP-aCGH can fine mapping the region of CNVs by high resolution from the whole genome-wide,which does contribute to limit the zones for finding pathogenic region and candidate genes,as well as to offer a technology platform for investigating about the correlation of phenotype and genes or CNVs.

Objective To investigate the influence of anti-anxiety and depression treatment on quality life and plasma B-type natriuretic peptide (BNP) for chronic heart failure (CHF) patients.Methods Eighty-two patients with CHF were divided into control group and intervention group by random digits table method with 41 cases each.After admission,the patients were given conventional anti-heart failure treatment.The patients in intervention group received the standard therapy in combination with lorazepam,with depression plus fluoxetine,and psychological treatment.After treatment,the patients were evaluated by symptom checklist (SCL-90),determined the plasma BNP level and heart function indexes,and recorded the length of stay,number of hospitalization,and so on.Results After treatment the intervention group somatization,interpersonal barriers as well as depression,anxiety,hostility,terrorism,paranoia,psychotic and other emotions significantly alleviated,there were statistical differences compared with before treatment and after treatment of control group (P ＜ 0.05).There were no statistical differences in heart function indexes before treatment between the 2 groups (P ＞ 0.05).After treatment in control group,only the left ventricular ejection fraction was significantly higher than that before treatment,there was statistical difference (P ＜ 0.05).After treatment in intervention group compared with before treatment and after treatment in control group,the heart rate and left ventricular early diastolic and late peak velocity ratio were significantly lower [(71.0 ± 10.9) times/min vs.(82.5 ± 12.4) and (77.3 ± 10.1) times/min,1.26 ± 0.28 vs.1.38 ± 0.21 and 1.31 ± 0.33],left ventricular ejection fraction was significantly higher [(50.9 ± 6.9)％ vs.(39.2 ± 7.4)％ and (43.4 ± 7.8)％],there were statistical differences (P ＜ 0.05).The plasma BNP level was downtrend after treatment in the 2 groups,compared with the control group,after 5 d of treatment in intervention group was significant decrease,the changes was slowly after 14 d of treatment.There were statistical differences in plasma BNP level in 5,7,10 d of treatment between the 2 groups (P ＜ 0.05).The length of stay,number of hospitalization and fatality rate in intervention group were lower than those in control group [(6.0 ± 1.4)d/time vs.(10.0 ± 2.1) d/time,(1.6 ± 0.5) times/year vs.(3.2 ± 0.8) times/years,0 vs.9.8％ (4/41)],there were statistical differences (P ＜ 0.05).Conclusion Anti-anxiety and depression treatment can significantly improve cardiac function in CHF patients,shorter hospital stays,and improve quality of life.

Aim:To develop a practical chiral CE method for the quantitative determination of the unwanted enantiomer[( R )-enantiomer]presented in zolmitriptan. Methods:The background electrolyte was 20 mmol/L sodium dihydrogenphosphate solution with 1% S-β-CD,adjusted to pH 3.50 with phosphoric acid. A fused-silica capillary(60 cm×50 μm ID,effective length 51.5 cm)was used at 20 ℃ for the separation. The applied voltage was -30 kV. The samples were loaded by hydrodynamic injection(50 mbar pressure,6 s). UV was measured at 220 nm. Results:Zolmitriptan and its chiral impurity were baseline resolved ( R s=6.66). The linearity was good over the concentration range from 4 to 80 μg/mL( r =0.999 8) of ( R )-enantiomer. The injection precision (expressed as CV%) was 2.83%. The average recovery was 99.97%( n =9). The limit of detection was 1.5 μg/mL. The host-guest complex binding constants were 964 and 905 mol-1 for ( R )-enantiomer and zolmitriptan,respectively. Conclusion:The method is suitable for the determination of ( R )-enantiomer in zolmitriptan and binding constants of zolmitriptan enantiomers to S-β-CD.

Objective To detect the serum levels of angiopoietin-like protein 2 (ANGPTL 2) and vascular endothelial growth factor (VEGF) in patients with rheumatoid arthritis (RA),and to analyze their value in the differential diagnosis of RA in order to clarify whether they are synergetic in the pathogenesis of RA.Methods Seventy-seven patients with RA and 25 subjects for heath check-up in the same period were selected.According to the relevant clinical information,the RA patients were divided into three subgroups:high disease activity group [(DAS28)＞5.1],moderate disease activity group (3.2＜DAS28 ≤5.1),low disease activity group (DAS28 ≤3.2).Enzyme-linked immunosorbent assay (ELISA) was used to masure the levls of ANGPTL2 and VEGF of those groups.The data were analyzed by t test,Mann-Whitney U test,Kruskal-Wallis H test and Speraman correlation analysis.Results ① The serum levels of ANGPTL2 and VEGF in RA patients were significantly higher than those in control group [(5.3±1.1) ng/ml vs (3.3±0.9) ng/ml,Z=-6.644;(106±30) pg/ml vs (93±12) pg/ml,Z=-4.4115,P＜0.05].② Serum level of ANGPTL2 in RA patients with high disease activity was higher than low disease group and healthy control group {[5.7(1.3)] ng/ml vs [4.5(0.3)]ng/ml,H=4.257,P＜0.05;[5.7 (1.3)] ng/ml vs [3.3 (0.9)] ng/ml,H=7.639,P＜0.05}.Serum level of VEGF in RA patients with high disease activity was higher than low disease group and healthy control group {[116(37)]pg/ml vs [96(8)] pg/ml,H=3.579,P＜0.05;[116(37)] pg/ml vs [92.90(12.24)] pg/ml,H=5.698,P＜0.05].③ There was a positive correlation between the serum level of ANGPTL2 and DAS28 score,erythrocyte sedimentation rate,swollen joint count and tender joint count (rDAS28=0.703,rESR=0.311,rSJC=0.503,rTJC=0.670,P＜0.05),so was VEGF (rDAS28=0.553,rESR=0.260,rSJC=0.399,rTJC=0.538,P＜0.05).④ In RA patients,there was a positive correlation between ANGPTL2 and VEGF (r=0.853,P＜0.05).Conclusion ANGPTL2 might coordinate with VEGF and be involved in the immune process of RA,and it might be regarded as one of the reference indexes of RA activity,so that it may contribute to access and monitor the condition of RA.

Objective To observe pre-syncopal limited tolerance and cardiovascular responses to head-up tilt combined with lower body negative pressure (HUT/LBNP) following exposure to head-down tilt (HDT, -1 Gz). Method Exposures to HUT/LBNP (-60 mmHg) in control session (without preceding 30 s -1 Gz treatment) and in simulated push-pull effect (PPE) session (with preceding 30 s -1 Gz treatment) were performed in 8 healthy adults. The changes of hemodynamic parameters were monitored by electrical impedance instrument during the experiments. Result The mean endurance time in presyncopal symptom limited HUT/LBNP in control session and in simulated PPE session were 8.4±2.1 min and 4.5±2.4 min, respectively, the two means were significantly different (P< 0.01). In simulated PPE session, as compared with baseline, heart rate (HR) during HDT was significantly lowered (P<0.01), while stroke volume (SV) and cardiac output (CO) were increased significantly (P<0.01). During HUT/LBNP, the increased percentage (relative to baseline) of HR in PPE session was lower than these in control session (P<0.05); the decreased percentages of SV and CO during HUT/LBNP in PPE session were both higher than those in control session (P<0.05). During HUT/LBNP, arterial pulse pressure (PP) of control session was significantly decreased than the value of baseline value (P<0.05); Total peripheral resistance (TPR) of PPE session was significantly increased than baseline value (P<0.05). Conclusion Tolerance time before the appearance of presyncopal symptoms during HUT/LBNP decreases and cardiovascular responses to HUT/LBNP are impaired, preceding exposure to HDT.