Humans ; Integrative Medicine ; Medicine, Chinese Traditional

Objective: To establish a loop-mediated isothermal amplification (LAMP) assay for rapid detection of the West Nile virus (WNV). Methods: WNV genome (position nt 1 021 to nt 1 240) was synthesized by a PCR-based gene synthesis method. The synthetic fragments included 6 pairs of LAMP primer recognizing 8 primer sites of WNV genome. The LAMP gene amplification was carried out using a real-time PCR system at 63°C for 60 min, then the amplification was terminated at 80°C after 2 min. The amplification products were observed by agarose gel electrophoresis. The sensitivity and specificity of LAMP assay were compared with those of conventional PCR. Results: The LAMP assay took less than 20 min, and the amplification product took on a ladder-like electrophoresis pattern. The sensitivity of LAMP assay was 10-fold higher than that of conventional PCR, and the detection limit of LAMP was 9.23 copies/μl. The specificity of WNV-specific LAMP assay was demonstrated by the negative amplification results from dengue virus and Japanese encephalitis virus, both were closely related members of the Flavivirus family. Conclusion: LAMP assay is rapid, cost-effective, highly sensitive and specific in detecting genes of interest, and is of great significance for WNV surveillance, especially for grass root units and on-sport surveillance.

Objective: To establish a single-tube nested multiplex-PCR assay for rapid detection and typing of Dengue viruses for multiple infections with different serotypes. Methods: A pair of outer universal primers designed for all the four Dengue virus serotypes were used to amplify the mixed RNA of 1-4 dengue viral serotypes by one-step RT-PCR, and the products were used as template for nested multiplex PCR using four pairs of serotype-specific primers in the same reaction tube. The sensitivity and specificity of single-tube nested multiplex PCR assay amplifying from the mixed 1-4 serotype dengue viral RNA were subsequently compared with those amplifying from the single serotypes dengue viral RNA. Results: By optimizing the reaction condition, four specific fragments (482,119,290,and 389 bp) were successfully amplified from the mixed RNA of 1-4 serotypes dengue viruses in single tube by single-tube nested multiple-PCR. Its sensitivity and specificity amplifying from the mixed RNA of 1-4 serotypes dengue viruses were similar to those amplifying from the single serotype dengue viral RNA. The detection limit of nested multiple-PCR was 66. 068 copies/μl. Conclusion: Single-tube nested multiple-PCR method is simple, rapid, sensitive, and specific for detecting and typing dengue viruses, and it is valuable for detecting and typing of the clinical multiple infections.

Objective: To elucidate the genetic characteristics and deduced protein variation of nonstructural protein (NS) gene of the novel influenza virus A/H1N1 in 2009 pandemic. Methods: The sequence of NS gene of A/H1N1 viruses isolated in North America, Europe, and Asia during 1930-2009 were downloaded from NCBI database. MEGA4.0 software and NJ method were used for sequence alignment, protein sequence alignment, and the phylogenetic tree construction. Results: The NS genes of novel influenza virus A/H1N1 in 2009 pandemic were originated from A/swine/H1N1 virus of 2005-2007; they shared a high homology of 97.5%-97.6%. There was an obvious evolutionary relationship between the NS genes of novel A/H1N1 virus and those of the influenza A/swine/H1N1 viruses isolated in North America from 1930 to 2007. No obvious changes were found in the amino acid sites for the antagonistic function of NS1 against the host antiviral capacity among these viruses. Conclusion: The NS gene of novel influenza virus A/H1N1 in 2009 pandemic might evolve from swine A/H1N1 influenza viruses isolated in the United States. The antagonistic function of NS1 against the host antiviral capacity is not changed.

Humans ; Myocardial Reperfusion Injury ; prevention & control ; Translational Medical Research

Cervical Intraepithelial Neoplasia ; diagnosis ; pathology ; surgery ; Colposcopy ; Conization ; Electrosurgery ; Female ; Humans ; Hysterectomy ; methods ; Neoplasm Regression, Spontaneous ; Uterine Cervical Neoplasms ; diagnosis ; pathology ; surgery ; Vaginal Smears

Animals ; Cytoprotection ; Humans ; Ischemic Preconditioning ; Reperfusion Injury ; prevention & control ; therapy

ObjectiveTo investigate the role of vascular endotheliar growth factor(VEGF),chemokine receptor 4(CXCR4) and stromal cell-derived factor-1(SDF-1) in Wilms tumor as well as their relationship with clinical features by examining the expressions of VEGF,CXCR4 and SDF-1.MethodsThirty cases of Wilms tumor samples and 12 cases of adjacent kidney tissue were collected from the First and the Third Affiliated Hospital of Zhengzhou University from May 2003 to May 2008.Thirteen boys and 17 girls aging from 4 months to 7 years old of whom were 22 favorable histologic types and 8 unfavorable histologic types.By means of cytoplasm to dye buffy for masculine cell,staining intensity and percentage of positive tumor cells serve as the judgment criteria for being positive or negative.All tissues would be tested by hematoxylin and eosin stain at the same time.ResultsThe positive expression rate of VEGF,CXCR4 and SDF-1 in Wilms tumor were 63.33%,70.0% and 53.33%.Those in adjacent normal kidney tissue were 25.0%,8.3% and 16.7%.The 2 groups were statistically significant(Pa

Objective To explore influential factors and features of relapse of presenting with an initial episode of acute central nervous system inflammatory demyelinating disease(IDDCNS) in children.Methods By group-control study,20 relapsing patients with IDDCNS compared with 50 single episoding patients with IDDCNS whose case history longer than 0.5 year,whose age at onset,gender,preceding infection or immunization,neurologic syndromes at presentation,therapeutic response,and final neurologic outcome after initial episode were analyzed by mono-factor analysis and multi-factor nonconditional Logistic regression model.Results 1.The risk of relapse increased in the near future,75.0% relapsed before 6 months.There were 18 cases diagnosed multiple sclerosis(MS),1 case neurooptic myelitis and 1 case immaturity myelitis in relapsing patients.There were 39 cases diagnosed acute disseminated encephalomyelitis in single episoding patients.2.Mono-factor analysis:8 factors were not statistically significant,including age at onset,gender,seizure,palsy,conscious disturbance,ataxia,natural relief,sequelae after the first attack;6 factors were statistically significant,including infectious or vaccination agents before episode,fever,visual disorder,multiple neurological signs,positive pathologic reflex,with marked improvement to cortical hormone.3.In the multi-factor stepwise regression analysis,visual disorder(OR=42.50)and positive pathologic reflex(OR=8.666)were the independent influencing factors,multiple neurological signs(OR=0.068)was independent protecting factor.Conclusions The rate of relapse in IDDCNS is lower than abroad and most relapse before 0.5 year,the main diagnosis of relapsing case is MS.When visual disorder and positive pathologic reflex presenting in the fist episode of IDDCNS,the opportunity of relapse will increase,and will decrease when multiple neurological signs presenting.

0.05 ).But the difference of positive ratio between 1-year-old group was significant in 6 cases of B19 DNA positive (P0.05).Conclusion There are no markedly association between kawasaki disease and human parvovirus B19 infection.