Objective To screen serum proteomic marker of hepatic echinococcosis, establish a diagnotic model of serum protein fingerprint patterns, and evaluate its clinical application for hepatic echinococcosis. Methods Serum samples from 68 patients with hepatic echinococcosis matched with 73 controls composed of 33 patients with liver diseases other than hepatic echinococcosis and 40 healthy people were collected. All subjects were divided into training group (37) and testing group (67). Serum protein profiling of patients with hepatic echinococcosis and controls were detected using surface enhanced laser desorption/ionization time of flight mass spectrometry(SELDI-TOF-MS) and weak cation exchange protein chip(WCX2). Peak intensities were compared, in the training group, between 37 patients with hepatic echinococcosis and 37 controls, 5 patients with HCE and 5 patients with HAE, and 8 patients with hepatic echinococcosis before and after operation, respectively. ZJU-Protein Chip Data Analyze System(ZJU-PDAS) was used for data analysis and the model of serum protein fingerprint patterns was build by support vector machine (SVM). The sensitivity and specificity of the model for diagnosis of hepatic echinococcosis were verified by blind method on samples of testing group. Results There were nine different protein peak spectra between hepatic echinococcosis group and control group, of which eight protein peak spectra decreased in patient group, their relative molecular mass were 1044, 1047, 1073, 1075, 1338, 6453, 6649, 8714 m/z, respectively, while one protein peak spectrum(5651 m/z) increased(P < 0.05). The sensitivity,specificity, positive predictive and negative predictive value of the model validated by blind method were 77.4% (24/31), 66.7% (24/36), respectively. There were two different protein peak spectra between HCE group and HAE group, Their relative molecule mass were 8716 and 2751 m/z, respectively (P < 0.05). Six different proteins were detected from pre-operation group and post-operation group. Their relative molecular mass were 1297, 1505, 1525, 1534, 5921, 5941 m/z, respectively(P < 0.05). Conclusions It is a successful way to screen serum proteomic marker in patients with hepatic echinococcosis by SELDI-TOF-MS and Bio-informatics, and the marker has a potential clinical value in diagnosis and judging prognosis of hepatic echinococcosis.

Objective To evaluate the effect of myocardial viability on left ventricular function after elective revascularization in patients with myocardial infarction by 99Tcm-MIBI and 18F-FDG dual-isotope simultaneous acquisition (DISA) myocardial perfusion-metabolic imaging. Methods Ninety-one patients clinically confirmed of myocardial infarction underwent DISA imaging. Based on the results of echocardiography, the patients were divided into heart failure group (group A) and normal cardiac function group (group B). After PCI, left ventricular function was measured by echocardiography in 1, 3 and 6 months. The t-test and χ2-test were used to compare the difference between the two groups using SPSS 13.0. Results The average number of diseased segments by myocardial perfusion imaging was 9.8±3.5 and 5.4±2.6 in groups A and B, respectively (t=6.87, P<0.01). The average number of diseased segments by myocardial metabolic imaging was 7.5±3.4 and 4.6±2.8 in groups A and B, respectively (t=4.46, P<0.01). There were 173 segments with viable myocardium (173/458: 37.8%) in group A and 188 segments with viable myocardium (188/307: 61.2%) in group B (χ2=40.61, P<0.001). The summed perfusion score (SPS), summed metabolism score (SMS) and summed difference score (SDS=SMS-SPS) were 28.43±11.86 vs 21.36±9.54, 20.17±8.52 vs 15.19±5.74 and 0.39±3.17 vs -12.72±4.55, respectively in groups A and B (t=3.15, P<0.01; t=3.32, P<0.01; t=15.59, P<0.01). The mean change of LVEF (ΔLVEF) and the mean change of left ventricular end-diastole dimension (ΔLVEDd) of the patients with more than 4 viable myocardial segments in group A were significantly more than those in group B( (12.81±2.62)% vs (5.90±1.91)%, t=16.33, P<0.001; (-13.13±4.20) mm vs (-7.75±2.31) mm, t=6.86, P<0.001). However, the ΔLVEF and ΔLVEDd of the patients with less than 4 viable myocardial segments in group A were significantly less than those in group B (t=3.25, P<0.01; t=4.92, P<0.001). Conclusion The amount of viable myocardium in infarct myocardium is an important factor for left ventricular function recovery after elective revascularization.

Objective To evaluate the protective effect of Yersinia pestis capsular antigen F1 and recombinant rV270 on mice after immunization with them.Methods According to body weight,40 female Balb/c mice aged 6 to 8 weeks were randomly divided into four experimental groups(Fl-10 μg + aluminum adjuvant,F1-20 μg + aluminum adjuvant,rV-10 μg + aluminum adjuvant,and rV-20 μg + aluminum adjuvant) and a control group,8 in each group.Mice in experimental groups were immunized with the natural antigen F1 and recombinant antigen rV270 adsorbed to 25％ aluminum adjuvant and the control group was immunized with the same amount of aluminum adjuvant.Each mouse was immunized at the hind leg muscle with 100 ml immunizing agent,then a booster immunization was done once on the 21st day after the first immunization.The blood of all mice was collected on the 8th week after the first immunization,serum antibody titers were detected by ELISA and the data of antibody titers were analyzed by t test for comparison between groups.At the same time the mice were injected subcutaneously with 2000-fold LD50 of Yersinia pestis virulent strain 141,after 14 days,the protective effect of immunization was analyzed.Results The control group did not produce antibody.Antibody geometric mean titers (GMT) of the F1-10 mg + aluminum adjuvant and F1-20 mg + aluminum adjuvant groups were 1 ∶ 30443.9,and 1 ∶21527.8,respectively,and compared between the two groups,the difference was not statistically significant (t =1.1282,P ＞ 0.05).The GMTs of the rV-10 μg + aluminum adjuvant and rV-20 μg + aluminum adjuvant groups were 1 ∶ 13957.3 and 1 ∶18100.9,respectively,and compared between the two groups,the difference was not statistically significant(t =0.9408,P ＞ 0.05 ).After subcutaneous injection with Yersinia pestis virulent strain 141,all mice died in the control group but all survived in the experimental group.Conclusion The immune activity of natural antigen F1 and recombinant rV270 is high,which can be used as the main component of subunit vaccine in the plague subunit vaccine study.

OBJECTIVETo explore the effect of Wnt signaling suppression on proliferation of non small cell lung cancer to gefitinib, and its related mechanisms.

METHODSPC9 and PC9/AB2 cells of both gefitinib sensitive and resistant were treated with different concentrations of gefitinib, and the proliferation index was measured using CCK8 kit. The members of Wnt signaling pathway were detected by Western blot. Dual luciferase reportor gene assay (TOP Flash) was used to document the transcriptional level of β-catenin. β-catenin siRNA was transfected into PC9/AB2 cells to suppress the Wnt signaling transcription, followed by treatment with different concentrations of gefitinib. Western blot was then used to detect the expression of EGFR and its downstream signaling after inhibit the expression of β-catenin.

RESULTSTreating with different concentrations of gefitinib, the resistance of PC9/AB2 cells to gefitinib was significantly increased (P < 0.05). The members of Wnt signaling expressed at higher level in PC9/AB2 cells than in PC9 cells (t = 24.590, P = 0.000). TOP Flash examination showed that the endogenous transcriptional activity of Wnt signaling was higher in PC9/AB2 cell than that in PC9 cell (t = 4.983, P = 0.008). Compared with the negative control group, apoptotic rate and sensitivity to gefitinib significantly increased in interfered group (P < 0.05). The expression of p-ERK1/2 significantly decreased after Wnt signaling suppression, although other proteins showed no significant alterations.

CONCLUSIONSuppressing the activity of Wnt signaling can partly reverse the celluar resistance to gefitinib in non small cell lung cancer.

Antineoplastic Agents ; administration & dosage ; pharmacology ; Apoptosis ; Carcinoma, Non-Small-Cell Lung ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Dose-Response Relationship, Drug ; Drug Resistance, Neoplasm ; Humans ; Lung Neoplasms ; metabolism ; pathology ; Mitogen-Activated Protein Kinase 1 ; metabolism ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Phosphorylation ; Quinazolines ; administration & dosage ; pharmacology ; Wnt Signaling Pathway ; drug effects ; beta Catenin ; metabolism

OBJECTIVETo study the distribution of genomovars and microevolution of Yersinia pestis in the Qinghai-Tibet Plateau.

METHODSPrimer pairs targeting the twenty-two different regions(DFRs) were designed for detecting the presence or deletion of each DFR in 297 strains isolated from the Qinghai-Tibet Plateau.

RESULTS9 genomovars, i. e. Genomovar 1, 5, 6, 7, 8, 10, 11, new type and Ype-ancestor were identified in the Marmota himalayana plague focus of the Qinghai-Tibet Plateau. Among these genomovars, genomovar 5,8 and 10 were dominant types. The total rate of the three genomovars was 80.6% (204/253) and the genomovars in different regions were different. All of 44 strains of Y. pestis in the Microtus fuscus plague focus of the Qinghai-Tibet Plateau belonged to genomovar 14.

CONCLUSIONThe distribution of genomovars of Y. pestis in the Qinghai-Tibet plateau had remarkable characteristics geographically. Based on the distribution of genomovars of Y. pestis, the routes of transmission and microevolution of Y. pestis were proposed.

Biological Evolution ; China ; Geography ; Humans ; Plague ; transmission ; Yersinia pestis ; genetics

Objective Throush identify biochemical characteristics and virulence factors of 2 strains suspected Yersinia pestis(Y.pestis)isolated from the dead Marmota himalayana(M.himalayana)to confirm the nature epidemic focus in Dege County,Sichuan Province.Methods Y.pestis was analyzed by specific staining and shape,culturing characteristics,splitting-test by bacteriophage,test of biochemical characteristics and glycolysis ability,virulence factors,virulence,nutritional requirement,plasmid,genetic test and genetic type. Results The tested strains were Gram staining bacilus.The main biochemical characteristics were Arabinose(+)、 Rhamnose(-),Maltose(+),Melibiose(-),Glycerol(+),Denitrification(+).The virulence factors with FI+.VW+, Pgm+,Pst I+;and with the common 6.0×106,45.0×106,65.0×106 plasmids,also with the virulence-relative plasmid gene.Both their absolutely lethal dose(LD100)in mice were 50 bacteria.The nutritional requirement appeared which were depended on Phenylalanine and Methionine.With the Genomovar 5 genotype characteristics of M.himalayana plague foci of Qinghai-Tibet plateau.The difference between tested strains and Yersinia pseudotubercuosis on the 3 different culture medium was obvious.The tested strains had a Y.pestis' specific 3a fragment,Pst I and FI-Ag,at 22 ℃,the strains could be split by bacteriophage completely.Conclusions According to the diagnostic criteria of plague in China,the 2 suspected strains isolated from Dege County,Sichuan Province ale confirmed as Y.pestis.both with powerful virulenceand with the characteristics of the Y.pestis of M.himahtyana in Qinghai-Tibet plateau plague natural focus.

Objective:Through phosphatidylinositol 3-kinase（PI3K）/protein kinase B （Akt）/mammalian rapamycin target protein （mTOR） signaling pathway， explore the effect of Zhigancao Tang on myocardial ischemia-reperfusion injury（MIRI）The role and mechanism of arrhythmia（ventricular tachycardia and ventricular fibrillation）. Method:The 72 SD rats were randomly divided into sham operation group，model group， Zhigancao Tang low，medium and high dose group（11.43，22.86，45.72 g·kg^-1），Wenxin granule group（2.43 g·kg^-1），continuous drug intervention for 10 days. Two hours after the last administration，the MIRI model of rat was prepared by ligating the left anterior descending coronary artery，and the changes of electrocardiogram were recorded. After successful modeling，blood and heart tissue were collected to detect the content of creatine creatine（CK），lactate dehydrogenase（LDH）and aspartate aminotransferase（AST）in the serum, the enzyme-linked immunoassay（ELISA） method was used to detect cardiac troponin（CtnI）content, immunohistochemical detection of myocardial PI3K，Akt，mTOR expression. Western blot was used to detect the myocardial autophagy-related protein microtubule-associated protein 1 light chain 3（LC-3），autophagy markers Beclin1 and PI3K/Akt/mTOR signaling pathway related protein expression and phosphorylated p-PI3K，p-Akt，p-mTOR levels. Result:In model group， 100% of ventricular tachycardia and 91.67% of ventricular fibrillation occurred. Compared with sham operation group， the serum levels of CK，LDH，AST，and CtnI in the model group were significantly increased（P<0.01），PI3K，Akt，mTOR AOD values in myocardial tissue were significantly increased （P<0.01），the relative expression of the ratio of LC3-Ⅱ/LC3-Ⅰ and Beclin1 was significantly increased（P<0.01），p-PI3K/PI3K，p-Akt/Akt，and p-mTOR/mTOR were significantly reduced （P<0.01）. Compared with model group， the incidence of ventricular tachycardia and ventricular fibrillation in the high-dose Zhigancao Tang group was significantly reduced （P<0.01），and the duration was the shortest compared with other administration groups（P<0.01）， CK，LDH，AST level and CtnI content were significantly reduced（P<0.01）， the expression of PI3K，Akt and mTOR of Zhigancao Tang group was significantly decreased with increasing dose（P<0.01）， the expression of LC-3 and Beclin1 was accompanied by Zhigancao Tang increase of each dose group of soup had different degrees of decrease （P<0.01），while the expression ratio of PI3K/Akt/mTOR-related protein was significantly increased （P<0.05，P<0.01）. Conclusion:Pretreatment of Zhigancao Tang can reduce the abusually elevated cardiac enzymes CK, LDH, AST and CtnI, inhibit excessive autophagy of cells, and up-regulate the expression of PI3K, Akt and mTOR, indicating that the anti-MIRI arrhythmia effect of Zhigancao Tang may be related to the PI3K/Akt/mTOR signaling pathway.