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 evaluate the value of intraduetal ultrasonography (IDUS) in diagnosing biliary and pancreatic disorders. Methods The findings by endoscopic retrograde cholangiopancreatography (ERCP) and IDUS from 19 patients with suspected biliary and pancreatic disorders from July 2006 to August 2007 in our hospital were analyzed retrospectively. Results Of the 19 patients, 17 had obstructive jaundice (including 6 eases of cholangiocarcinoma, 2 pancreatic adenocareinoma, 2 gallbladder carcinoma, 2 chole-docholithiasis with bile duct stricture, 2 autoimmune pancreatitis, 1 papillary adenocarcinoma, 1 papillary adenoma, and 1 sclerosing cholangitis) and 2 intraduetal papillary mueinous tumor (IPMT). The diagnosis was confirmed by surgery and pathological findings in 11 patients. The diagnostic accuracy of ERCP and IDUS was 73. 7% (14/19) and 84. 2% (16/19), respectively, and that of ERCP combined with IDUS was 89. 5% (17/19). The sensitivity and specificity of ERCP to differentiate benign bile duct strictures from ma-lignant ones were 100. 0% (11/11) and 83.3% (5/6), respectively; and those of IDUS were 100. 0%(11/11) and 100. 0% (6/6), respectively. The sensitivity and specificity of ERCP in diagnosing cholan-gioeareinoma were 83.3% (5/6) and 60% (3/5), respectively; and those of IDUS were 100. 0% (6/6) and 40. 0% (2/5), respectively. Conclusion Combination of ERCP with IDUS can improve the diagnostic accuracy of pancreaticobiliary disorders. Additionally, IDUS shows higher sensitivity and specificity in differ-entiation between benign and malignant bile duct strictures, but it is still difficult to identify the etiologic factors of malignant bile duet strictures by IDUS.

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.

Objective To evaluate the effects of mechanical stretch on pentraxin-3(PTX-3)mRNA and protein expression in human alveolar epithelial cells (A549 cells).Methods The human lung epithelial adenocarcinoma cells A549(A549 cells)were purchased from cell biology laboratory,Tongji Medical College,Huazhong University of Science and Technology.The cultured A549 cells were inoculated on collagen Ⅰ BioFlex plates and divided into 5 groups(n=3 wells each):group Ⅰ normal control;groupⅡsham mechanical stretch;group Ⅲ mechanical stretch;groupⅣsiRNA and groupⅤ siRNA+mechanical stretch.In group Ⅲ the cells underwent square cyclic mechanical stretch for 4 h using the Flexercell Systcm.In group Ⅳ the cells were transfected with chemosynthetic PTX-3 specific siRNA by RNAi technique.In group Ⅴ at 24 h after being transfected with PTX-3 siRNA the cells underwent mechanical stretch for 4 h.In groupⅡ mechanical stretch of the cells were prevented by Flexstep.The expression of PTX-3 mRNA in the cells was detected by real-time PCR and the expression of PTX-3 protein in the culture media was determined by Western blotting.Apoptosis of the cells was measured bv flow cytometry(Beeten-Dickinson,USA).Results PTX-3 mRNA and protein expression was signlficantly up-regulated by mechanical stretch in groupⅢand decreased by transfection with siRNA in group Ⅳand Ⅴ as compared with group Ⅰ andⅡ(P＜0.05 or 0.01).The apoptosis ratio was significantly higher in group Ⅲ and Ⅴ than in groupⅡ and was significantly lower in group Ⅴ thanin group Ⅲ(P＜0.01).Conclusion Mechanical stretch can up-regulate PTX-3 mRNA expression in A549 cells.

Objective To evaluate the prognostic value of lateral lymphadenectomy for advanced lower rectal cancer. Methods A total of 96 consecutive patients who underwent total mesorectal excision and lateral lymphadenectomy for advanced lower rectal cancer were retrospectively evaluated by using therapeutic index. A product of the frequency of lymph node metastasis and the cancer-related 5-year survival rate. Results The incidences of lymph node metastasis in mesorectal area, superior rectal artery area, inferior mesenteric artery area and lateral area was 21%,13%.10%and 15%respectively.The 5-year survival rate of patients with metastasis to the mesorectal area, superior rectal artery area, inferior mesenteric artery area and lateral area was 35%,25%,20%and 36%respectively.The therapeutic indexes for total mesorectal excision and lateral lymphadenectomy were 7.4 and 5.4,which were much higher than that of lymphadenectomy of the superior rectal artery area(3.3)and inferior mesenteric artery area(2.0).Local recurrence occurred in 19%(18 out of 96 cases)patients with advanced lower rectal cancer. Local recurrence rate of patients with positive lateral lymph node metastasis was 64%(9 of 14 cases),whereas it was 11%(9 of 82 cases)in those without lateral lymph node metastasis. The difference between these two groups was statistically significant(x2=22.308,P=0.000).Kaplan-Meier survival analysis showed significant improvements in median survival(80.9±2.1)m,95%CI:76.7～85.1 m vs(38±6.7)m,(95%CI:24.8～51.2 m)for patients with negative lateral lymph node metastasis over patients with lateral lymph node metastasis(log-rank,P=0.000). Conclusion The current results indicate that lateral pelvic lymphadenectomy could significantly reduce loeal recurrence and effectively improve the survival of patients with advanced lower rectal cancer. Except for total mesorectal excision, lateral pelvic lymphadenectomy should be followed in the surgery of pailents with advanced low rectal cancer.

Objective To research the mechanism or pathway through which Substance P(SP) inhibits r-aminobutyric acid(GABA) activated currents in bullfrog dorsal root ganglion(DRG) neurons.Method The experiment was conducted on freshly isolated bullfrog dorsal root ganglion neurons using a whole-cell patch-clamp technique.Results SP could caused a slow inward current when SP was applied to DRG neurons;SP could inhibits GABA-activated currents;The inhibition could be reduced largely when protein kinase C(PKC) inhibiter,1-(5-isoquinolinesulphorry)-2-methylpiperazine(H-7), was dialyzed in cell body.Conclusion The SP ton inhibit GABA-activated currents through protein kinase C.

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 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 .