ObjectiveTo explore the application of the Structure Equation Model on validity assessment of the evaluation indicator system,we studied Medical Capital Development Fund projects in 2007 to evaluate their construct validity.MethodsPart of the evaluation results (1200 items) were divided into two parts randomly.PART1 was used for Exploratory Factor Analysis (EFA) to explore the Indicator System' structure,and formed model F1.Confirmatory Factor Analysis (CFA) was used to assess the construct validity of model F1.With considerations of Modification Index(MI) and reality,F1 was modified into F3.At last,we take PART2 as the sample to confirm F3.ResultThe results of model F3 fit indices showed a better fit.All the path coefficients,estimated load factors and other parameters were statistically significant.ConclusionStructural Equation Model is professional,reliable and satisfactory for the evaluation of indicator system's contract validity.

Objective:To investigate the training needs of nursing students during their practice in the emergency department, and to provide evidence for further improvement and development of the training.Methods:A total of 279 nursing students from 24 hospitals in Guangdong Province and 4 undergraduate universities running nursing specialty were investigated by convenience sampling. The contents of the questionnaire included general information and emergency training needs, including teaching contents and teaching methods.Results:Among the surveyed nursing students, 97.13% (271 students) believed it was very necessary to learn first aid knowledge and skills, and 85.66% (239 students) were very interested in learning first aid knowledge and skills. The score of "acute and critical skills" in the emergency training needs was the highest (88.91 points), and the score of "department rules and regulations" was the lowest (76.08 points). Univariate analysis showed that there were significant differences in gender, willingness to engage in nursing work in the future, interest in learning first aid knowledge and willingness to learn first aid knowledge independently by network ( P < 0.05). The three teaching methods ranked the highest in the survey of emergency teaching methods were situational teaching, problem-based learning and action teaching method. Conclusion:Nursing students have a strong demand for training during the emergency internship. Clinical nursing administrators and educators should combine the internship demand of nursing students and the teaching outline of medical colleges to carry out the training arrangement during the emergency practice, so as to meet the learning needs of nursing students during the emergency practice.

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 determine the effect of calcitonin gene-related peptide (CGRP) on triggering receptor expressed on myeloid cells-1 (TREM-1) in the lipopolysaccharide (LPS)-induced macrophages and its signal transduction pathway. Methods The levels of TREM-1 mRNA in the macrophages were observed by reverse transcription-polymerase chain reaction (RT-PCR), and flow cytometry was performed to detect TREM-1 protein expression levels in the macrophages. Results CGRP had no regulating effect on the expression of TREM-1 in the macrophages; LPS could up-regulate macrophages to express TREM-1; CGRP increased TREM-1 mRNA expression in LPS-induced macrophages in dose and time-dependent manner; CGRP increased TREM-1 protein expression in LPS-induced macrophages, which could be partially reversed by H-7 or H-89 (P<0.05). Conclusion CGRP can regulate the LPS-induced macrophages synthesis and secretion of TREM-1, and the intracellular signal transduction pathway is related to PKA and PKC.

Objective To investigate the immunogenicity and cross protective effects of two novel HCV DNA vaccines in a mice model.Methods Two self-replicating alphavirus vector-based HCV DNA vaccines, pSCK CE1E2Y and pSCK H155, were constructed based on the genes encoding the structural pro-teins (Core, E1 and E2) and structural and NS3 fusion proteins (Core, E1 , E2 and NS3) of a HCV strain isolated from a Chinese patient (genotype 1b, Hebei strain), respectively.Western blot analysis was per-formed to detect the expression of fusion antigens.The BALB/c mice were intradermally immunized with the recombinant DNA vaccines by using electroporation.The immune responses induced in mice and the cross protective effects of the recombinant DNA vaccines were evaluated.Results The DNA vaccines effectively expressed the target antigens in vitro.The antigen-specific antibody responses and specific T cell immune re-sponses were induced in mice by the immunization of replicative DNA vaccines.However, no effective cross protection was provided by either of the DNA vaccines in the surrogate challenge model based on a recombi-nant heterologous HCV (JFH1, 2a) vaccinia virus strain.Conclusion Although no effective cross protec-tion was observed, both of the two replicative DNA vaccines could induce strong humoral and cellular im-mune responses against multi-target antigens of HCV strains.This study has paved the way for further inves-tigation on the development of novel HCV vaccines.

Objective To investigate the development strategy of novel T cell based vaccine against HCV infection.Methods BALB/c mice were primed with pSCK-based DNA vaccine and boosted with type 5 adenoviral vector-based vaccine, which expressed the structural proteins ( Core, E1 and E2) de-rived from a Chinese HCV patient (genotype 1b, Hebei strain).Enzyme linked immunospot assay (ELIS-POT) and intracellular cytokine staining ( ICS) were used to analyze the elicited antigen-specific immune re-sponses and the efficacy of cross-protection.Results Immunization of mice with the prime-boost vaccination strategy elicited stronger T cell immune responses against multiple HCV antigens than using the DNA vac-cines alone, especially the IFN-γ-secreting T cell responses against E1 protein as indicated by ELISPOT as-say.ICS data indicated that the prime-boost regimen elicited more TNF-α-producing CD4+and IFN-γ-produ-cing CD8+T cells against E1 protein and high levels of IFN-γ-producing CD4+and CD8+T cells against E2 protein in comparison with immunization with DNA vaccines.Moreover, the prime-boost vaccination was ca-pable of eliciting effective cross-protection in a surrogate challenge model based on a recombinant heterolo-gous HCV (JFH1, 2a) vaccinia virus.Conclusion The prime-boost vaccination using DNA and rAd5-based vaccine expressing HCV structural antigens induced significant cellular immune response and cross-protection in mice, suggesting the possibility of using it as a promising T cell based vaccine against HCV in-fection.

To develop a safe and broad-spectrum effective hepatitis C virus (HCV) T cell vaccine,we constructed the recombinant adenovirus-based vaccine that carried the hepatitis C virus truncated NS3 and core fusion proteins. The expression of the fusion antigen was confirmed by in vitro immunofluorescence and western blotting assays. Our results indicated that this vaccine not only stimulated antigen-specific antibody responses,but also activated strong NS3-specific T cell immune responses. NS3-specific IFN-γ+ and TNF-α+ CD4+ T cell subsets were also detected by a intracellular cytokine secretion assay. In a surrogate challenge assay based on a recombinant heterologous HCV (JFH1,2a) vaccinia virus,the recombinant adenovirus-based vaccine was capable of eliciting effective levels of cross-protection. These findings have im- portant implications for the study of HCV immune protection and the future development of a novel vaccine.
Adenoviridae ; genetics ; metabolism ; Animals ; CD4-Positive T-Lymphocytes ; immunology ; Cross Protection ; Female ; Genetic Vectors ; biosynthesis ; genetics ; Hepacivirus ; genetics ; immunology ; Hepatitis C ; immunology ; prevention & control ; virology ; Humans ; Interferon-gamma ; immunology ; Mice ; Mice, Inbred BALB C ; Recombinant Proteins ; administration & dosage ; genetics ; immunology ; Viral Core Proteins ; administration & dosage ; genetics ; immunology ; Viral Hepatitis Vaccines ; administration & dosage ; genetics ; immunology ; Viral Nonstructural Proteins ; administration & dosage ; genetics ; immunology

Objective To compare the effects of three kinds of Oncomelania hupensis RNA extraction methods,namely a modified SDS method,TRIzol reagent method,and CTAB method,so as to obtain an economical and efficient method for RNA extraction from O. hupensis. Methods The modified SDS method,TRIzol reagent method and CTAB method were applied to ex-tract the RNA from O. hupensis. A nucleic acid protein analyzer was used to measure the concentration and purity of RNA. The yields were calculated by the concentration of the products. The purity was indicated by A260/A280 and A260/A230. The quality of RNA was inspected by 1% agarose gel electrophoresis. The β-acting gene was selected as the target gene for RT-PCR analysis. Re-sults The RNA yields obtained by using the three kinds of extraction methods were significantly different(F = 16895.85,P <0.01)according to the analysis of variance. The LSD test showed that the yields obtained by using the modified SDS method were the highest,and those obtained by the CTAB method were the lowest. The purity of RNA extracted by the CTAB method was su-perior to that by the other two methods,and the A260/A280 and A260/A230 ratios of the CTAB method were in the range from 1.8-2.0 and 2.0-2.2. The A260/A230 ratios of the other two methods were both lower than 2.0. The RNA extracted by the modified SDS meth-od had the better integrity. The electrophoresis results showed that the 28S rRNA band,18S rRNA band and 5S rRNA band were clear,and there was no obvious smear between each band. The RNA obtained by the TRIzol reagent method had no 28S rRNA band,and that obtained by the CTAB method had no 28S rRNA and 5S rRNA bands. The β-acting gene of the RNA ex-tracted by all the three methods could be amplified by RT-PCR. The costs and time-consuming of the modified SDS method were less than those of the other two methods. Conclusion The modified SDS method is an economic and efficient method,and it is suitable for extracting the RNA of O. hupensis,especially for large sample preparation.

Objective To investigate the characteristics of coronary vessel re-endothelialization after placement of drug-eluting stents (DES), and to provide clinical evidence for the double anti-platelet treatment. Methods Optical coherence tomography (OCT) was performed in 43 patients in 1 year after DES implantation. Characteristics of re-endothelialization and percentage of neointimal coverage of stent struts were evaluated by OCT. Results The rate of stent struts intimal coverage was 90.70%, and the remain was lack of endothelial coverage; The ratio of neointimal thickness (NIT) between 0-99, 100-199 and above 200 microns was 19.92%, 37.55% and 42.53%, respectively. The rate of neointimal coverage was higher and the degree of neointimal hy-perplasia was more extensive in patients with DM and in patients with ACS than those of patients without DM and of patients with stable angina pectoris. Conclusion One year after stent placement, most of the stent struts were covered with neointima and few struts obtained poor coverage of endothelial. DM and ACS may be impact factors for the progress of re-endothelialization after DES placement.

Objective To characterize the immunogenicity in gene immunization of the conserved regions of hepatitis C virus(HCV) based on different delivery strategies. Methods We first constructed a novel DNA vaccine encoding a fusion gene(from partial NS3 and Core) of HCV. Then we compared different protocols based on naked DNA injection twice or DNA injection with gene electrotransfer(GET) in BALB/c mice. The immune response was measured by antibody ELISA and by IFN-gamma ELISPOT. Results Our data showed that a protocol based on intradermally injection of DNA with optimal GET induced the strongest humoral and cellular immunity, and DNA with GET induced a substantially higher anti-NS3/Core T cell re-spoase than naked DNA injection. Conclusion Our data suggest that DNA vaccines encoding NS3/Core fu-sion protein of HCV immunized by the present strategy could merit further study in the context of future prophylactic and therapeutic HCV T cell based vaccines.