ObjectiveMatching pursuit algorithm(MAP),for its good parametric characterization,was applied in epileptic electroencephalography(EEG) to study time-frequency distribution.MethodsSimulation experiment of time-frequency analysis was carried out to verify the matching pursuit algorithm s superiority on frequency resolution and parametric characterization.Fourier transform,Wigner-Ville distribution and matching pursuit algorithm were applied to the time-frequency analysis on normal EEG and epileptic EEG to study epileptic discharge in the time-frequency plane and the results were compared.ResultsSimulation results showed that the matching pursuit algorithm obtained a better time-frequency distribution.Distributions of epileptic EEG and normal EEG had significant difference in time-frequency plane.ConclusionTime-frequency analysis based on matching pursuit can better reveal the EEG characteristics.

The resting 12 lead electrocardiograms (ECG) of 45 patients with Wolff-Parkinson-White syndrome and 30 normal persons were assessed by five criterias for locating the accessory pathway. Golden criteria was established by site of accessory pathway confirmed during epicardial mapping and absent accessory pathway from normal person. Compared five criterias of EGG with golden criteria, the idiosyncrasy of every criteria was 100% and the sensitivity of Rosenbaum's cri-terias, Gallagher's, WHO's, Lindsay's, Reddy's were respectively 93. 3% , 71. 1% , 64. 4% , 60% , 53. 3%. The preliminary feature for locating the accessory pathway of ECG was put forward from advantages of five criterias and observation of this report data.

The morphologic changes of human hepatocaranoma transplanted in nude mice afler photodynamic therapy (PDT) with sulfonated aluminum phthalocyanine (ALSPQ were studied The cancer cells after PDT with ALSPC showad vacuolar degeneration and separation from each other. The endothelium of blood vessel was damaged and hemorrhage occured Under electron microscope the endoplasmic reticulum enlarged, mitochondria swelled, perinudear space dilated, and chromosome aggregated and condensed. The membrane of nuclei and cells broke off and tumor cells died at last The endothdial cells had the same changes mentioned above. This result suggests that ALSPC has photodynamic effect on the human hepatocellular carcinoma tranaplanted in nude mice. And the membrane system of cells is probably the main target.

Objective To elucidate the molecular basis on the differences of infectivity in infected cells between Sindbis virus(SINV:YN87448 virus)and Sindbis like virus(SINLV:XJ-160 virus).Methods Compare the E(glycoprotein)gene sequence and secondary structure of YN87448 virus and XJ-160 virus by bioinformatics analysis.Analyze the contribution of E gene to the biological differences between SINV and SINLV by constructing recombinant virus.Results By bioinformatics analysis,YN87448 virus and XJ-160 virus have the same genomic structure,which has 11 717 nt and 11 626 nt respectively.There are 82 amino acid differences between E gene of these two viruses,and showed scattered distribution.The main peak is basically the same for the hydrophobic of the E gene protein,but in some region existing small differences.The recombinant virus which exchanged the E gene of XJ-160 virus with YN87448 virus totally showed the biological character of YN87448 virus,either in the showing time of CPE,plaque forming time and plaque diameter,or in expression of functional proteins.Conclusion E gene plays a major role in the differences of infectivity in infected cells between SINV and SINLV,this result provide the molecular biological evidences for elucidating the biological differences between SINV and SINLV.

Objective:To obtain the high-efficiency expression of the biological active rabies virus nucleoprotein in the prokaryotic expression system.Methods:This experiment uses codon optimization technology to re-encode the nucleoprotein gene of rabies virus CTN-1 strain, artificially synthesize the full-length gene and clone it into pET-43.1a prokaryotic expression vector, induced expression in BL21 (DE3) strain of Escherichia coli( E. coli), and used Western blot to detect its reactogenicity. Results:The results showed that after induction, SDS-PAGE electrophoresis analysis showed that an obvious expression band appeared at a molecular weight of 50×10 3, which was consistent with the expected protein band size. Among them, the E. coli concentration A600 is about 0.5, and the expression yield is the highest (about 32.3%) when induced at 37℃ for 5 h. Nucleoprotein expression product is mainly inclusion body when it is expressed in large quantities. After purification by Ni 2+ chelating chromatography, the purity of the target protein can reach over 95%. The purified product was identified by Western blot and positively reacted with the sera of mice immunized with rabies vaccine, indicating that the prokaryotic expression of the CTN-1 strain nucleoprotein has biological activity. Conclusions:This experiment successfully established a high-efficiency expression method for the nucleoprotein of the CTN-1 strain in the prokaryotic expression system, and obtained high-purity target protein, which provides a basis for further clinical diagnosis and preparation of new vaccines.

To establish a cell-based rapid luciferase suppression assay for high-throughput screening (HTS) anti-alphaviruses compounds screening, which could cause viral encephalitis, raise the social issues associated directly with public health and huge economic burden to the society. The Gaussia luciferase assay system was used for HTS model for identifying inhibitors of labeled virus XJ160-GLUC. The decreased 50% GLUC activity inhibition ratio was deemed to be the screening positive index. The reaction system in this model was optimized, and the reliability of the model was evaluated. For HTS model's optimization, cells were infected with XJ160-GLUC at an MOI of 0.025 PFU/cell. The supernatant treated with compounds 48h were collected for GLUC expression detection. In the model, Z' factor was up to 0.71, demonstrating that HTS assay for identifying inhibitors that target all aspects of the viral life cycle of XJ160-GLUC was stable and reliable. After screening 8080 compounds (five-in-one), 341 positive samples were selected, and the positive rate was 4.2% with a cutoff at 50% inhibition. Then 1705 compounds were screened subsequently and the positive rate was 1.1% with obtaining 19 positive compounds. These results will lay the foundation for finding the anti-alphaviruses' drug targets.
Alphavirus ; drug effects ; genetics ; metabolism ; Animals ; Antiviral Agents ; pharmacology ; Drug Evaluation, Preclinical ; methods ; Genes, Reporter ; High-Throughput Screening Assays ; methods ; Luciferases ; genetics ; metabolism

Objective To investigate the surgery-related complications and risk factors of ileocolic Crohn's disease (CD).Methods The retrospective case-control study was conducted.The clinicopathological data of 52 patients with ileocolic CD who underwent surgery at the Peking Union Medical College Hospital from January 2010 to April 2016 were collected.Observation indicators:(1) surgery-related complications,(2) risk factors analysis of surgery-related complications:gender,age of onset,preoperative body mass index (BMI),course of disease,smoking history,history of appendectomy,perianal lesions,oral ulcer,C-reactive protein (CRP),erythrocyte sedimentation rate,disease behavior,short crohn's disease activity index (sCDAI),preoperative amino salicylic acid therapy,preoperative hormone therapy,preoperative antituberculosis therapy,preoperative immunosuppressive agents therapy,preoperative biologic agents therapy,emergency operation,surgical method and ileocolic anastomosis method,(3) follow-up.The follow-up using outpatient examination and telephone interview was performed to detect recurrence of disease up to August 2016.Measurement data with normal distribution were represented as-x ± s.The univariate analysis was done using the chi-square test,and multivariate analysis was done using the Logistic regression model.Results (1) Surgery-related complications:of 52 patients,12 had postoperative complications.Four patients complicated with wound infection had good healing of the wound after debridement and dressing change.Of 4 patients with abdominal infection,3 were improved by anti-infection symptomatic treatment and 1 die of septic shock at postoperative day 1.One patient with intestinal obstruction had a smooth recovery after open adhesiolysis.One patient with intestinal fistula discharged from hospital due to a critical condition under families' requestion.One patient with acute cholecystitis and 1 with acute pancreatitis were respectively improved by conservative treatment.(2) Risk factors analysis of surgery-related complications:theresult of univariate analysis showed that sCDAI and emergency operation were the factors infecting surgery-relatedcomplications of ileocolic CD (x2 =6.299,8.494,P ＜ 0.05).The result of multivariate analysis showed that sCDAI was an independent factor infecting surgery-related complications of ileocolic CD [OR =2.716,95％ confidence interval (CI):1.216-6.066,P ＜ 0.05].(3) Follow-up:all the 52 patients were followed up for 5-76 months with a median time of 39 months.During the follow-up,15 had recurrence of diseases and then underwent medical treatment.Conclusions Patients with ileocolic CD are easily complicated with wound infection and abdominal infection in the active period,and sCDAI is an independent factor infecting surgery-related complications of ileocolic CD.

Objective To establish a CVS-11 pseudovirus particles ( pp)-based assay for detec-tion of neutralizing antibody against rabies virus. Methods An improved rapid fluorescence focus inhibition test ( RFFIT) for detection of neutralizing antibody against rabies virus ( RVNA) was established based on the CVS-11 pseudovirus expressing a luciferase reporter gene. Forty-six human serum samples were analyzed with the improved RFFIT and the results were compared with those by using standard RFFIT. Moreover, the improved RFFIT was used to detect the titers of RVNA in 91 serum samples collected from pet dogs and pet-breeders in Beijing. Results The coincidence rate of the improved RFFIT and the standard RFFIT was 100% regarding to the analysis of 46 human serum samples and 5 negative reference serum samples. Moreo-ver, the RVNA titers of all serum samples obtained with CVS-11 pseudovirus-based assay showed a signifi-cant high correlation with those obtained with standard RFFIT (n=46, r=0. 94, P<0. 000 1). All of the 91 serum samples collected from pet dogs and pet-breeders in Beijing were positive for RVNA as indicated by the improved RFFIT with a mean titer of 33. 01 IU/ml. Conclusion We established an improved RFFIT based on the CVS-11 pp expressing luciferase reporter gene, which might be used as a reliable alternative RFFIT for measuring RVNA titer. Analysis of the 91 serum samples collected in Beijing with the improved RFFIT showed that all samples were positive for RVNA.

Objective To evaluate the importance of plasma MicroRNAs in early diagnosis of acute myocardial infarction (AMI).Methods 24 patients with AMI as the test group and 20healthy volunteers as the control group were enrolled in this study.Plasma levels of microRNA-1,microRNA-133a,microRNA-208a and microRNA-499 were detected by quantitative real time polymerase chain reaction (qRT-PCR) at 3 h,6 h,12 h,24 h,48 h,72 h after the onset of AMI.Results Plasma microRNA-1 level was greatly increased and reached the peak at 3 h after AMI,then was decreased gradually to normal level at 72 h after AMI.Plasma microRNA-133a level was significantly elevated at 6 h after AMI,reached peak at 12 h after AMI,then was decreased to normal level at 48 h after AMI.Plasma microRNA-1 and microRNA-133a levels were correlated with cTnI expression.The peak time of microRNA-1 was earlier than that of cTnI,while the peak time of microRNA-133a was the same as that of cTnI.Conclusions Increased circulating microRNA-1 and microRNA-133a may serve as potential and novel biomarkers for early diagnosis of AMI.

Objective To investigate the value of plasma miR-22 in diagnosis of idiopathic pulmonary arterial hypertension ( IPAH ),and its role of regulation mechanisms in the pathogenesis of the disease.Methods Circulating miR-22 levels of IPAH patients and healthy controls were evaluated by RTPCR.The silico analysis of targets for miR-22 was taken, and followed by eGFP reporter assay for verification of predicted target gene Myc binding protein (MYCBP). Results Compared with healthy controls,the expression of plasma miR-22 in IPAH patients was significantly decreased (P ＜ 0.01 ).The area under curve (AUC) of ROC curve was 0.744.MYCBP was a real target of miR-22 confirmed by silico analysis and eGFP reporter assay. Conclusions The expression of plasma miR-22 was significantly decreased in IPAH patients,and it could serve as a potential biomarker for diagnosis.The miR-22 might be involved in the pathogenesis of the disease through promoting its target gene MYCBP to activate the c-Myc pathway.