0.05).Thirty-four cases of newly diagnosed children with CR rate was 88.2% (30 cases).Which the expression of PRAME gene,WT1 gene were both positive and negative groups of children with CR rates were 62.5% and 100.0%,there was no significant difference between the 2 groups(P

A simple, rapid and sensitive colorimetric reverse-transcription loop-mediated isothermal amplification (RT-LAMP) was developed for rapid detection of Middle East respiratory syndrome coronavirus (MERS-CoV). The method employed six primers that recognized sequences of a nucleocapsid gene for amplification of nucleic acids under isothermal conditions at 63 degrees C for 60 min. Products were detected through a LA-320c Loopamp Turbidimeter (real-time RT-LAMP) or visual inspection of color change by pre-addition of Hydroxynaphthol Blue dye (visual RT-LAMP). Specificity of RT-LAMP was validated by detection of several human coronaviruses and common respiratory viruses. MERS-CoV real-time RT-LAMP had a linear correlation (R2) of 0.995 at 10(3)-10(6) copies. The limit of detection of real-time RT-LAMP, visual RT-LAMP and quantitative real-time PCR was 500, 1000 and 100 copies/reaction, respectively. The established RT-LAMP assay was demonstrated to be a rapid screening tool for MERS-CoV infection, and could be suitable in resource-limited clinical sites and for field studies.
Coronavirus Infections ; virology ; DNA Primers ; genetics ; Humans ; Middle East Respiratory Syndrome Coronavirus ; classification ; genetics ; isolation & purification ; Nucleic Acid Amplification Techniques ; methods ; Reverse Transcription

Objective To construct and express ribonuclease-resistant virus-like particles containing the RNA fragmengts of MERS-CoV N gene.Methods The coat protein and maturase gene of E.coli bacteriophage MS2 was amplified by PCR,then the gene was cloned into pET32a to construct the intermediate vector pET32MS.The gene fragments harboring MERS-CoV N gene and beta-actin was cloned into the downstream of pET32MS to construct the prokaryotic expression vector p32MS-EMC-Beta.The recombinant plasmid p32MS-EMC-Beta was transformed into E.coli BL21 (DE) competent cells and induced with IPTG.The virus-like particles were obtained after purification.RNase digestion test and stability test were carried out to observe the stability of the particles.Results The RNase-resistant virus-like particles which was able to express the gene fragments containing MERS-CoV N gene and beta-actin were successfully produced and were shown to be stored stable for 30 days at 37℃.Conclusion The virus-like particles with high safety and stability can be used as positive standards and quality controls in the application of MERS-CoV detection.

Objective To study the safety and efficacy of continuous intravenous infusion of fentanyl in laser photocoagulation of retinopathy of prematurity (ROP).Methods From March,2014 to January,2015,ROP infants hospitalized for laser photocoagulation in Guangdong Women and Children hospital were randomly (using envelope method)assigned into the fentanyl groupand the control group.In the fentanyl group,the patients were given fentanyl combined with topical anesthesia,while onlytropical anesthesiain the control group.Premature infant pain profile (PIPP)scores,heart rate,mean artery pressure and complications within 3 days after operation were recorded. The concentration of epinephrine, norepinephrine and cortisol in the blood were measured before and after the operation.Student′s t test,non-parameter rank and chi-square test were used to compare the differences between the two groups.Results A total of 82 infants were enrolled in the study,42 in the fentanyl group and 40 in the control group.In the fentanyl group,11 .9% infants had maximum PIPP score ≥6 and 70.0% in the control group,the difference was statistically significant (P <0.05).In the fentanyl group,the PIPP score at the beginning of operation, the maximum PIPP score during operation and the PIPP score at the end of operation were 2.0,3.0and 1 .5, respectively.In the control group,these scoreswere 8.0,8.0and 8.0 respectively.The differences were statistically significant (P <0.05 ).No significant differences existed between the concentration of epinephrine,norepinephrine and cortisol before and after operationin the fentanyl group.However,these concentrations were elevated after the operation than before the operation in the control group (P <0.05).The incidence of complications within three days after operation was 19.0% in the fentanyl group and 40.0% in the control group,and the difference was statistically significant (P <0.05 ).Conclusion Comparing with topical anesthesia,fentanyl combined with topical anesthesia has lower pain scores,less stress responses and fewer complications during ROP laser photocoagulation.Fentanyl combined with topical anesthesia is a safe and effective analgesic method during ROP laser photocoagulation.

Small cell lung cancer is the most common primary neuroendocrine malignancy of the lung and is characterized by high malignant degree,rapid doubling time,easy metastasis in early stage and poor prognosis.Accurate staging of small cell lung cancer can formulate personalized therapeutic plans and improve the prognosis of patients.PET/CT can obtain metabolism and anatomical images of the whole body in one scan and improve the diagnostic accuracy and integrity.PET/CT has been widely applied to clinical practice now.PET/CT will play a more and more important role in diagnosis,staging,treatment and prognosis assessment of patients with small cell lung cancer.The value of PET/CT in staging and treatment of small cell lung cancer was reviewed in this article.

Given the Ebola outbreak in West Africa and the risks of spread to other regions, a rapid, sensitive and simple method for the detection of the Ebola virus (EBOV) is of great significance for the prevention and control of Ebola. We developed a simple colorimetric isothermal multiple self-matching initiated amplification (IMSA) for rapid detection of the Zaire subtype of the Ebola virus (EBOV-Z). This method employed six primers that recognized seven sites of the EBOV-Z nucleoprotein gene for amplification of nucleic acids under isothermal conditions at 63 degrees C for 1 h. Amplification products were detected through visual inspection of color change by pre-addition of hydroxyl naphthol blue dye. Relative sensitivity was validated by detection of serial tenfold dilutions of virus-like particles containing the partial EBOV-Z nucleoprotein gene and mock clinical sample. Specificity of IMSA was validated by detection of the plasma of 30 healthy volunteers, the dengue virus, and Japanese encephalitis virus. IMSA had comparable sensitivity to Reverse transcription-quantitative polymerase chain reaction (RT-qPCR), and cross-reaction with human plasma or other viruses was not observed. Reverse transcription-isothermal multiple self-matching initiated amplification (RT-IMSA) was also evaluated and compared in parallel with the commercial RT-qPCR kit for detection of EBOV-suspected samples of human blood in Sierra Leone. Sensitivity and specificity of the RT-IMSA was 91.4% and 100%, respectively. These data suggest that RT-IMSA is a valuable tool for the detection of the EBOV with the distinct advantages of simplicity and low cost compared with RT-qPCR.
Colorimetry ; methods ; DNA Primers ; genetics ; Ebolavirus ; genetics ; isolation & purification ; Hemorrhagic Fever, Ebola ; diagnosis ; virology ; Humans ; Nucleic Acid Amplification Techniques ; methods

The Flanders virus (FLAV) is a number of family Rhabdoviridae ,contains a single‐stranded ,negative‐sense vi‐ral RNA .Here we describe a molecular detection method developed for fast measurement of FLAV based on Taqman RT‐PCR method .In this study ,FLAV specific primers and probe were designed based on the FLAV L gene sequences published in GeneBank .Quantitative standard curve of FLAV TaqMan PCR was also successfully established .The specificity and stability test showed that the system is specific and the coefficient variables were all less than 1 .7% .Quantitative standard curve based on the genomic copy was drawn ,and the lowest detectable limit (LOD) of system was 100 copies/PCR ,with higher sensitivity and stability than that of the conventional RT‐PCR assay targeting the same gene .

BACKGROUNDAccumulating evidence indicates that both innate and adaptive mechanisms are responsible for the postnatal development of the mammalian visual cortex. Most of the studies, including gene expression analysis, were performed on the visual cortex during the critical period; few efforts were made to elucidate the molecular changes in the visual cortex during much earlier postnatal stages. The current study aimed to gain a general insight into the molecular mechanisms in the developmental process of the rat visual cortex using microarray to display the gene expression profiles of the visual cortex on postnatal days.

METHODSAll age-matched Sprague-Dawley rats in various groups including postnatal day 0 (P0, n = 20), day 10 (P10, n = 15), day 20 (P20, n = 15) and day 45 (P45, n = 10) were sacrificed respectively. Fresh visual cortex from the binocular area (Area 17) was dissected for extraction of total RNA for microarray analyses. Taking advantage of annotation information from the gene ontology and pathway database, the gene expression profiles were systematically and globally analyzed.

RESULTSOf the 31 042 gene sequences represented on the rat expression microarray, more than 4000 of the transcripts significantly altered at days 45, 20 or 10 compared to day 0. The most obvious alteration of gene expression occurred in the first ten days of the postnatal period and the genomic activities of the visual cortex maintained a high level from birth to day 45. Compared to the gene expression at birth, there were 2630 changed transcripts that shared in three postnatal periods. The up-regulated genes in most signaling pathways were more than those of the down-regulated genes.

CONCLUSIONSAnalyzing gene expression patterns, we provide a detailed insight into the molecular organization of the developing visual cortex in the earlier postnatal rat. The most obvious alteration of gene expression in visual cortex occurred in the first ten days. Our data were a basis to identify new relevant candidate genes that control visual cortex development.

Lipase from Candida sp. 99-125 was immobilized by physical adsorption onto macroporous resins. The results showed that the nonpolar resin NKA was the best carrier used in low aqueous media. 98.98% of degree of immobilization can be achieved when the adsorption procedure was performed in the presence of heptane. The hydrolytic activity and the apparent activity recovery of lipase adsorbed on resin in heptane was 4.07 and 3.43 times higher than that of lipase adsorbed in sodium phosphate buffer, respectively. The catalytic properties of immobilized lipase for production of biodiesel in low aqueous media were studied. Immobilized lipase displayed the highest activity when the crude enzyme/resin weight ratio was 1.92:1 and the water content(water/oil weight ratio) was 15% at 40 degrees C under pH 7.4. As lipase was adsorbed on NKA in heptane to produce biodiesel, the batch conversion rate can reach 97.3% when a three-step methanolysis protocol was used. After 19 consecutive batches, the conversion rate remained 70.2%.
Candida ; enzymology ; Enzymes, Immobilized ; metabolism ; Gasoline ; Lipase ; isolation & purification ; metabolism ; Porosity ; Resins, Synthetic ; chemistry ; Soybean Oil ; chemistry

Objective To develop the technique to detect total core antigen of HCV(Total HCV-cAg) by Enzyme-Linked Immu-nosorbent Assay (ELISA) and apply it for clinical diagnosis. Methods 201 serum samples with anti-HCV antibody were detected total HCV-cAg after pre - treating the samples, then the sensitivity of results were compared with HCV RNA tests. Among them, 176 cases was determined by FQ-PCR, and 25 cases by RT-PCR for HCV-RNA. Results HCV RNA was found in sera from 88 of 201 samples (43.8%). Total HCV-cAg was positive in 71 (35.3%) of 201 samples . There was no significant difference between the detection rate of HCV RNA by PCR and total HCV-cAg by ELISA. Conclusion Detection of total core antigen of HCV is suitable to be used as to diagnose HCV in clinic.