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.

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

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.

Objective To investigate the effect of eosinophils(EOS)in asthmatic guinea pigs airway and explore the mechanism of antiasthma by inhaled arsenic trioxide. Methods The guinea pigs asthma models were established with ovalbumin(OVA)challenge method,2 groups received inhaled different doses of arsenic trioxide,and three control groups respectively received normal saline inhaled and high dose of arsenic trioxide by intraperitoneal injection and dexamethasone(DEX)intraperitoneally. EOS invaded and apoptosis with all of the groups were assessed after 7 days intraperitoneal injection or inhalation. Results Compared with group of inhaled normal saline, both EOS invaded and apoptosis in bronchus submuconsa were significantly different(P0.05)among groups of inhaled low dose arsenic trioxide [2.0 mg(kg?d)]and intraperitoneal injection with higher dose arsenic trioxide[5.0 kg/(kg?d)]and DEX[10 mg/(kg?d)]for these two parameters. Conclusions The mechanism of arsenic trioxide antiasthma with arsenic trioxide can decrease EOS amount in bronchial submucosa and accelerate EOS apoptosis, and relieve bronchial inflammation in asthma. For inhaled lower dose arsenic trioxide or intraperitoneal injection with high dose arsenic trioxide ,the effect of EOS is equivalent. The antiasthma effect of inhaled lower dose arsenic trioxide[2.0 mg/(kg?d)]is equivalent to intraperitoneal injection higher dose[5.0 mg/(kg?d)],and may be safe comparatively. The mechanism of antiasthma by inhaled arsenic trioxide is the same with DEX.

Objective To explore the effect of Renzhu Jianwei Granules(Granule of Semen Coicis and Rhizoma Atractylodis Macro- cephalae for strengthening the stomach)on the patients with precancerous lesions of gastric cancer(PLGC)and its mechanism. Methods The 58 patients of chronic atrophic gastritis(CAG)complicated with intestinal metaplasia(IM)and/or dysplasia(Dys) were randomized into treatment group and control group with 29 in each.The treatment group was administered Renzhu Jianwei Gran- ules while the control group was given Weifuchun Tablets.The numbers of positive cells of carcinoembryonic antigen(CEA)and cy- clooxygenase 2(COX-2)of gastric mueosa before and after treatment were observed.Results The scores of positive cells of CEA and COX-2 of gastric mucosa of both groups were greatly reduced after treatment(P0.05).Conclusion Renzhu Jianwei Granules can reduce CEA and COX-2 of PLGC patients,possibly being one of the mechanisms in treating PLGC.

Objective To investigate the clinical features of spinal muscular atrophy type Ⅱ.Methods The data of clinical manifestation,laboratory data, onset, diagnosis, and rehabilitation of 53 outpatients suffering from spinal muscular atrophy type Ⅱ (SMA-Ⅱ) were analyzed. Results Among 53 patients with SMA-Ⅱ confirmed by molecular genetic tests, 27 patients were male while 26 were female. The mean age was 3.35 years (ranged 0.75~7.8 years), and the age receiving gene diagnosis was 17.27 months. Only 15% had a family history,and abnormalities were found in 23% patients' mothers during pregnancy. 83.4% of them had water choke cough, while 87% expectoration weakness. None of them had cough assist machines and had regular spirometry monitoring. 53% of the patients took semi-liquid diet, however,none of them used stomach tube. Symmetrical flaccid paralysis was so remarkable, even muscle strength of lower limbs in 87.7% of these patients were only grades 1-2. 92% had scoliosis, while 83% had tendon contracture. EMG showed extensive neurogenic changes. All children did not accept normal pre-school education, and 85% patients did not accept formal rehabilitation. Conclusion Most of phenotype of SMA-Ⅱ for the children was similar and more severe in China. Most of the patient didn't get formal education, rehabilitation and care.

Objective To evaluate the therapy and turnover of Kawasaki disease(KD)with glucose-6-phosphate dehydrogenase(G-6-PD)deficiency.Methods Six hundred and twenty-four patients with KD were selected including 32 patients who had G-6-PD defected.The same dose intravenous immunoglobulin(IVIG)was used in all 624 patients(2 g/kg),but used Dipyridamole in 32 patients had G-6-PD deficiency which replaced the role of acetylsalicylic acid(ASA)after acute period.The coronary artery of these patients were checked and followed-up through echocardiograph.The turnover of 32 patients with G-6-PD deficiency and 356 case selected randomly from all the KD patients were compared.SPSS 10.0 software was used to analyze the data.Results In 32 cases of KD with G-6-PD deficiency,4 children had coronary aneurysm(12.5%).After 6-12 months follow-up,the coronary lesions were recovered in 62.5% children,improved in 21.9% children and not improved in 15.6% childern,which were not significantly different from all the KD patients(Z=-1.604 P=1.09).Conclusions IVIG and Dipyridamole are feasible in treating KD with G-6-PD deficiency.J Appl Clin Pediatr,2009,24(1):26-27

Behcet Syndrome ; complications ; Humans ; Male ; Middle Aged ; Myocardial Infarction ; complications