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Affected by the uncertainties caused by cable cutting and assembly errors, material performance dispersion, and truss deformation, the dual-surface accuracy of a mesh antenna cannot satisfy the design requirements. However, the existing methods can only adjust the accuracy of a single surface. This paper studies a method for adjusting coupled dual-surface mesh antennas considering uncertainties. Dual-reflector mesh antennas use both the front and rear reflectors for sensing, and both surfaces have shape accuracy requirements. First, an interval model updating method based on sensitivity analysis is proposed to obtain an updated interval parameters model group, which can accurately represent the relationship between adjustment cable length and surface accuracy. Then, an interval optimization function for the coupled dual-surface accuracy is established to get optimal adjustment amounts, which is solved by using the fast elitist nondominated sorting genetic algorithm (NSGA-II). Meanwhile, a kriging model is introduced to improve the optimization efficiency. Finally, the AstroMesh antenna is taken as an example to verify the validity of the proposed method. The results show that both the front and back net accuracies can be improved after adjustment, proving the proposed method’s feasibility.

STUDY DESIGN A generic qualitative design. OBJECTIVES To obtain a deeper understanding of the outcomes of spinal cord injury (SCI) peer mentorship programs delivered by community-based organizations. SETTING Peer mentorship programs of community-based SCI organizations METHODS: We interviewed 36 individuals who shared their experiences of SCI peer mentorship from the perspective of a peer mentee, peer mentor, or family member of a peer mentee/mentor, or staff of SCI community-based organizations. Interview data were analyzed using an inductive thematic analysis approach. RESULTS Four overarching themes with sub-themes were identified. (1) Positive outcomes for mentees such as understanding, emotional outlet/psychological support, inspiration/hope, and belonging. (2) Positive outcomes for mentors such as gaining gratitude, confidence, pride, and personal growth. (3) Reciprocity in positive/negative outcomes for mentors and mentees, such as shared learning and a lack of connection. (4) Negative outcomes for mentors such as impact of negativity, emotional toll, and time/energy demands. CONCLUSIONS Peer mentorship programs delivered by community-based SCI organizations are important, impactful resources for individuals with SCI who engage in these programs. These results provide insights into the variety of positive and negative outcomes linked with these programs.

As the main data transport protocol of delay/disruption-tolerant networking, Licklider transmission protocol (LTP) is developed to provide reliable and highly efficient data delivery over unreliable deep-space and interplanetary communication channels. For reliable data delivery, a checkpoint (CP) segment of LTP is sent, with a timer set (or, simply, CP timer) to check the arrival status of the entire data block at the receiver, and it is retransmitted upon the expiration of the CP timer prior to reception of an acknowledgment. Little work has been seen in analysis of the CP timer length for optimal performance of LTP in energy-efficient, reliable data delivery. In this article, a packet-level analysis of the CP timer length setting is presented for the reliable data delivery of LTP over unreliable deep-space communication channels based on realistic file transfer experiments conducted using a PC-based testbed. It is our intent to find an optimal CP timer setting so that the reliable file delivery in deep space is ensured with a minimum number of retransmission attempts. By this, the energy-efficient, reliable data delivery of LTP can be achieved in deep-space communications.

Peer mentorship is a flagship program utilized by Canadian community-based spinal cord injury (SCI) organizations. Through connecting trained SCI peer mentors with fellow adults with SCI, these programs help adults adapt and thrive following their injury. The objective of this meta-synthesis was to work with SCI community organizations and to identify outcomes of participating in community- or rehabilitation-based peer mentorship programs using an integrated knowledge translation approach.A meta-synthesis of 21 qualitative peer-reviewed studies and 66 community documents was conducted.A total of 87 outcomes of peer mentorship were identified.The outcomes of peer mentorship were grouped according to six higher-order themes: 1) Independence: enhanced self-sufficiency; 2) Personal growth: positive psychological changes; 3) Activities and participation: greater participation in activities and events; 4) Adaptation: adapting to life with disability; 5) Knowledge: obtaining new information, resources, and opportunities; and 6) Connection: developing and maintaining social relationship.The positive nature of the identified outcomes suggests that participating in peer mentorship can promote improved health and quality of life for adults with SCI. Furthermore, the integrated knowledge translation approach helped identify outcomes that were previously not examined within SCI peer mentorship research, thus providing important insight for future research.

Objective First, to examine whether participants reported changes in (1) leisure-time physical activity (LTPA) participation and social inclusion variables and (2) well-being outcomes before and after joining a community-based LTPA program for adults with physical disabilities. Second, to explore the longitudinal relationship between LTPA and the other aforementioned outcomes. Design A double baseline longitudinal design with measurements at 4-6 weeks (baseline 1) and immediately (baseline 2) before and 2 and 4 months after joining the community-based LTPA program. Setting Community. Participants Adults (N=43) with a physical disability who reported no cognitive impairment, were new members of the community-based LTPA program, and spoke English or French. Interventions A community-based physical activity program for adults with physical disabilities. Participants were provided an individualized exercise program and accessed the program at designated times during the week. Main Outcome Measures Primary: LTPA (LTPA Questionnaire for People with Spinal Cord Injury), participation (Patient-Perceived Participation in Daily Activities Questionnaire), and social inclusion. Secondary: depression severity, self-esteem, resilience, and life satisfaction. Results After joining the program, participants reported an increase in total LTPA (meanbaseline2, 177.80±211.32; mean2months, 299.31±298.70; mean4months, 288.14±292.14), moderate-to-vigorous LTPA (meanbaseline2, 83.95±123.95; mean2months, 142.00±198.38; mean4months, 163.23±182.08), and participation in health (meanbaseline2, 6.24±1.16; mean2months, 6.58±1.25; mean4months, 6.97±0.82) and family-related activities (meanbaseline2, 12.18±2.43; mean2months, 12.60±2.30; mean4months, 13.47±2.01). A significant increase (β=3.46, P .05). Conclusions The results support the role of a community-based LTPA program in increasing LTPA levels and enhancing participation in some activities among adults with physical disabilities.

Generic qualitative design. To explore how Chinese adults living with spinal cord injury (SCI) viewed the prospect of inpatient peer support programs within a rehabilitation setting. Hospital in China. A purposive sample of adult inpatients with SCI (N = 6) currently undergoing rehabilitation was recruited. Each participant was interviewed twice. Twelve interview transcripts were analyzed using a thematic method. Five higher-order themes were developed. First, participants had unique backgrounds and personal lives before and after their SCI and reported frustrations about their lives resulting from their SCI. Second, participants reported varying degrees of satisfaction with their rehabilitation and identified the facilitators and barriers to their rehabilitation. Third, their perspectives on peer support were shaped by their rehabilitation goals. For example, participants who solely focused on the recovery of physical functioning noted that peers could help to supplement existing rehabilitation programming by guiding their rehabilitation exercises. Participants who concentrated on their future lives believed peers could teach them new skills to facilitate their integration in the community. However, some participants felt they could not trust peers’ advice because peers are not healthcare providers. Fourth, peer support delivery options varied from online chat groups (i.e., WeChat), in-person conversations, and mentoring lectures. Finally, anticipated outcomes were related to obtaining practical and emotional support from peers, being motivated, and feeling understood. Participants harbored mixed views on potential use-value and necessity of hospital-based peer support programs, which could inform future utilization of SCI peer support within Chinese hospitals.

The umbrella cable-net structure is one of the most popular forms for deployable space reflectors and has been successfully applied to many in-orbit deployable antennas. The umbrella ribs o...

Metagenomic analysis through high-throughput sequencing is a tool for detecting both known and novel viruses. Using this technique, a novel circular single-stranded DNA (ssDNA) virus genome was discovered in respiratory secretions from a febrile traveler. The virus, named human respiratory-associated PSCV-5-like virus (HRAPLV), has a genome comprising 3,018 bases, with two major putative ORFs inversely encoding capsid (Cap) and replicase (Rep) protein and separated by two intergenic regions. One stem-loop structure was predicted in the larger intergenic region (LIR). The predicted amino acid sequences of the Cap and Rep proteins of HRAPLV showed highest identity to those of porcine stool-associated circular virus 5 isolate CP3 (PoSCV 5) (53.0% and 48.9%, respectively). The host tropism of the virus is unknown, and further study is warranted to determine whether this novel virus is associated with human disease.

Human infection with H7N9 virus has provoked global public health concern due to the substantial morbidity and mortality. Neuraminidase inhibitors (NAIs) are used as first-line drugs to treat the infection. However, virus quasispecies can evolve rapidly under drug pressure, which may alter various biological characteristics of virus. Using an in vitro evolution platform and next-generation sequencing, we found the presence of peramivir led to changes to the dominant populations of the virus. Two important amino acid substitutions were identified in NA, I222T and H274Y, which caused reduced susceptibilities to oseltamivir or both oseltamivir and peramivir as confirmed by enzyme- and cell-based assays. The NA-H274Y variant showed decreased replicative fitness at the early stage of infection accompanied with impaired NA function. The quasispecies evolution of H7N9 virus and the potential emergence of these two variants should be closely monitored, which may guide the adjustment of antiviral strategies.

We develop color code-based pyrosequencing with di-base addition for analysis of single nucleotide polymorphisms (SNPs). When a di-base is added into the polymerization, one or several two-color code(s) containing the type and the number of incorporated nucleotides will be produced. The code information obtained in a single run is useful to genotype SNPs as each allelic variant will give a specific pattern compared to the two other variants. Special care has to be taken while designing the di-base dispensation order. Here, we present a detailed protocol for establishing sequence-specific di-base addition to avoid nonsynchronous extension at the SNP sites. By using this technology, as few as 50 copies of DNA templates were accurately sequenced. Higher signals were produced and thus a relatively lower sample amount was required. Furthermore, the read length of per flow was increased, making simultaneous identification of multiple SNPs in a single sequencing run possible. Validation of the method was performed by using templates with two SNPs covering 37 bp and with three SNPs covering 58 bp as well as 82 bp. These SNPs were successfully genotyped by using only a sequencing primer in a single PCR/sequencing run. Our results demonstrated that this technology could be potentially developed into a powerful methodology to accurately determine SNPs so as to diagnose clinical settings.

Objective To express and purify two kinds of antigens of Coxiella burnetii (C. burnetii), the main outer membrane protein Com1 and the acute disease antigen A (adaA), in prokaryotic expression system and to validate the two recombinant antigens by mass spectrometry and identify their antigenicity. Methods The gene sequences encoding Com1 and adaA were separately synthesized and constructed into the prokaryotic expression vector pET-20b(+). The constructed vectors were transformed into E.coli BL21(DE3), and the recombinant proteins were induced by IPTG. The recombinant Com1 and adaA were purified by His affinity chromatography and identified by mass spectrometry. The immunoreactivity of the two antigens was identified by Western blot analysis using Q fever positive bovine serum. Results The expression vectors pET-20b(+)-Com1 and pET-20b(+)-adaA were constructed and the recombinant Com1 and adaA were expressed and purified in a soluble form. High-purity recombinant Com1 and adaA were obtained after purification, and the SDS-PAGE showed that their relative molecular masses were M

Yiyue Ge,1 Qiang Zhou,2 Kangchen Zhao,1 Ying Chi,1 Bin Liu,3 Xiaoyan Min,4 Zhiyang Shi,1 Bingjie Zou,2 Lunbiao Cui1 1Institute of Pathogenic Microbiology, Key Laboratories of Enteric Pathogenic Microbiology (Ministry of Health), Jiangsu Provincial Center for Disease Control and Prevention, 2Department of Pharmacology, Jinling Hospital, Medical School of Nanjing University, 3Department of Biomedical Engineering, Nanjing Medical University, 4Department of Geriatrics, The First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu, People’s Republic of China Abstract: Influenza virus infections represent a worldwide public health and economic problem due to the significant morbidity and mortality caused by seasonal epidemics and pandemics. Sensitive and convenient methodologies for detection of influenza viruses are essential for further disease control. Loop-mediated isothermal amplification (LAMP) is the most commonly used method of nucleic acid isothermal amplification. However, with regard to multiplex LAMP, differentiating the ladder-like LAMP products derived from multiple targets is still challenging today. The requirement of specialized instruments has further hindered the on-site application of multiplex LAMP. We have developed an integrated assay coupling multiplex reverse transcription LAMP with cascade invasive reaction using nanoparticles (mRT-LAMP-CIRN) as a sensor for the detection of three subtypes of influenza viruses: A/H1N1pdm09, A/H3 and influenzaB. The analytic sensitivities of the mRT-LAMP-CIRN assay were 101 copies of RNA for both A/H1N1pdm09 and A/H3, and 102 copies of RNA for influenza B. This assay demonstrated highly specific detection of target viruses and could differentiatethem from other genetically or clinically related viruses. Clinical specimen analysis showed the mRT-LAMP-CIRN assay had an overall sensitivity and specificity of 98.3% and 100%, respectively. In summary, the mRT-LAMP-CIRN assay is highly sensitive and specific, and can be used as a cost-saving and instrument-free method for the detection of influenza viruses, especially for on-site use. Keywords: multiplex, LAMP, gold nanoparticles, on-site detection, influenza virus

Considering the fatal human victims and economic loss caused by influenza virus infection every year, methodologies for rapid and on-site detection of influenza viruses are urgently needed. LAMP is the most commonly used nucleic acid isothermal amplification technology suitable for on-site use. However, for multiplex LAMP, differentiation of the amplicons derived from multiple targets is still challengeable currently. Here we developed a multiplex RT-LAMP assay for simultaneous amplification of three prominent subtypes of influenza viruses (A/H5, A/H7 and 2009A/H1). The amplicons were further identified by cascade invasive reaction and nanoparticle hybridization in separate target-specific detection tubes (referred to as mRT-LAMP-IRNH). The analytic sensitivities of the assay are 10 copies of RNA for all the three HA subtypes, and the specificity reached 100%. Clinical specimen analysis showed this assay had a combined sensitivity and specificity of 98.1% and 100%, respectively. Overall, the mRT-LAMP-IRNH assay can be used as a cost-saving method that utilizes a simple instrument to detect A/H5, A/H7, and 2009A/H1 influenza viruses, especially in resource-limited settings.

A novel avian-origin influenza A (H7N9) virus recently occurred in China and caused 137 human infection cases with a 32.8% mortality rate. Although various detection procedures have been developed, the pathogenesis of this emerging virus in humans remains largely unknown. In this study, we characterized serum microRNA (miRNA) profile in response to H7N9 virus infection using TaqMan Low Density Arrays. Upon infection, a total of 395 miRNAs were expressed in the serum pool of patients, far beyond the 221 in healthy controls. Among the 187 commonly expressed miRNAs, 146 were up-regulated and only 7 down-regulated in patients. Further analysis by quantitative RT-PCR revealed that the serum levels of miR-17, miR-20a, miR-106a and miR-376c were significantly elevated in patients compared with healthy individuals (p < 0.05). Receiver operating characteristic (ROC) curves were constructed to show that each miRNA could discriminate H7N9 patients from controls with area under the curve (AUC) values ranging from 0.622 to 0.898, whereas a combination of miR-17, miR-20a, miR-106a and miR-376c obtained a higher discriminating ability with an AUC value of 0.96. Our findings unravel the significant alterations in serum miRNA expression following virus infection and manifest great potential of circulating miRNAs for the diagnosis of viral diseases.

SUMMARYIn a large serosurvey performed in Gaobeidian city, China, 1032 children and adolescents aged 3–18 years were divided into three age groups: 3–6, 7–11, and 12–18 years. The geometric mean of IgG antibodies against pertussis toxin (anti-PT IgG) varied from 3·47 IU/ml (3–6 years) to 8·71 IU/ml (12–18 years) and the seroprevalence (⩾40 IU/ml) varied from 4·48% (7–11 years) to 11·76% (12–18 years). Infections were clustered in three secondary schools (schools 1–3). However, excluding secondary schools 1–3, the IgG PT levels, seroprevalence and rates of recent infections (>100 IU/ml) in secondary schools 4–7 (where infections were not clustered) were very close to those in the 3–6 and 7–11 years age groups. This suggests that the high seroprevalence and anti-PT IgG levels in adolescents in Gaobeidian city, China were caused by the clustered cases in certain secondary schools. Our results also suggested that pertussis was still circulating in China, and the reported incidence might be underestimated.

Highlights • Drug-resistant influenza viruses present a major public health problem. • We present an assay for the identification of resistance-conferring mutations. • We tested 26 clinical samples of an infectious H7N9 influenza virus for mutations. • The assay successfully identified a mixed population of E120/E120V associated with drug-resistant mutant. • Parallel testing of clinical specimens and cultured samples may avoid misinterpreting culture-induced mutations., A novel reassortant avian influenza A virus (H7N9) emerged in humans in Eastern China in late February 2013. All virus strains were resistant to adamantanes (amantadine and rimantadine), but susceptible to neuraminidase inhibitors (NAIs) (oseltamivir and zanamivir). One strain (A/shanghai/1/2013) contained the R294K substitution in the neuraminidase (NA) gene, indicating resistance to oseltamivir. Pyrosequencing has proven to be a useful tool in the surveillance of drug resistance in influenza A viruses. Here, we describe a reverse transcription (RT)-PCR assay coupled with pyrosequencing to identify the NA residues E120, H276, and R294 (N9 numbering) of H7N9 viruses. A total of 43 specimens (26 clinical samples and 17 isolates) were tested. Only one isolate containing the E120V heterogenic mutation was detected by pyrosequencing and confirmed by Sanger sequencing. However, this mutation was not detected in the original clinical specimen. Since virus isolation might lead to the selection of variants that might not fully represent the virus population in the clinical specimens, we suggest that using pyrosequencing to detect NAI resistance in H7N9 viruses directly from clinical specimens rather than from cultured isolates. No cross-reactions with other types of influenza virus and respiratory tract viruses were found, and this assay has a sensitivity of 100 copies of synthetic RNA for all three codons. The high sensitivity and specificity of the assay should be sufficient for the detection of positive clinical specimens. In this study, we provide a rapid and reliable method for the characterization of NAI resistance in H7N9 viruses.

Influenza A viruses present a significant threat to public health worldwide. High-affinity human scFv antibodies against a conserved epitope can potentially provide immunity to diverse viruses and protect against future pandemic viruses. A library of phage-displayed human scFv containing 6.0×10(8) members was generated from lymphocytes of H5N1 virus vaccinated individuals. Using the recombinant H5N1 virus hemagglutinin ectodomain (HA1), 4F5 scFv was identified with neutralizing activity against both clade 2 and 9 H5N1 viruses. In embryonated chicken eggs, the antiviral activity of 4F5 scFv conferred a 100% survival rate and at least a 62.5% survival rate against different clades of H5N1 viruses by pre-treatment and post-treatment, respectively. 4F5 scFv belongs to the VH-3-43 family according to the IMGT database, and a peptide (76)WLLGNP(81) containing half of an α-helix in HA1 was identified as the binding pocket. The conserved binding epitope of this novel broadly neutralizing scFv may become key in the design and implementation of vaccines or RNA interference against H5N1 viruses.

Acute diarrhea is a leading cause of childhood morbidity and mortality worldwide, but there have been few reports on the causative viruses associated with acute diarrhea among outpatient children in developing countries. This study was conducted to identify the viral agents in outpatient children with acute diarrhea in southeastern China.Eight hundred eleven outpatient children 5 years or younger with acute diarrhea were enrolled. Five enteric viruses were determined by enzyme-linked immunosorbent assay and multiplex reverse transcription-polymerase chain reaction for each stool specimen.At least 1 virus was detected in 353 (43.5%) of the subjects. The proportions of rotavirus, norovirus, sapovirus, adenovirus and astrovirus were 25.5%, 18.1%, 4.4%, 2.7% and 1.2%, respectively. G3P[8] was the most prevalent rotavirus strain. Mixed infections were observed in 65 cases, among which the most prevalent coinfection was rotavirus with other viruses (58 cases, 89.2%). Rotavirus and norovirus infections showed marked and opposing seasonal patterns. Mixed infection was significantly more common in children older than 1 year (12.2%) than in those younger than 1 year (7.1%) (P = 0.026). Clinically, rotavirus infection presented with a longer duration (4.3 ± 6.7 days) and higher frequency (5.9 ± 2.0 times/d) of diarrhea than any other viral infection. Vomiting was more common for mixed infections than for single infections (P = 0.010).All the 5 common etiologies were detected in this study, with rotavirus and norovirus being the 2 leading agents. Mixed viral infections were common in outpatient children with acute diarrhea, and rotavirus seemed to play a major role in mixed infections.

Pertussis is a highly contagious, respiratory disease associated with substantial morbidity and mortality. A rapid and reliable diagnostic method is essential for appropriate treatment and prevention. Expression profiles of circulating microRNAs (miRNAs) have been proven as new non-invasive biomarkers for infectious diseases. We aimed to investigated the serum miRNA profile in pertussis patients and explored its potential as a novel diagnostic biomarker for pertussis. Among 664 different miRNAs analyzed using a miRNA array, 50 were overexpressed and 81 were underexpressed in the serum of pertussis patients. Expression levels of seven candidate miRNAs were further evaluated by real-time qRT-PCR. A panel of five miRNAs (miR-202, miR-342-5p, miR-206, miR-487b, miR-576-5p) was confirmed overexpressed in pertussis patients (p

Hand, foot, and mouth disease (HFMD), mainly caused by coxsackievirus A16 (CVA16) and enterovirus 71 (EV71) infections, remains a serious public health issue with thousands of newly diagnostic cases each year since 2008 in China. The mechanisms underlying viral infection, however, are elusive to date. In the present study, we systematically investigated the host cellular microRNA (miRNA) expression patterns in response to CVA16 and EV71 infections. Through microarray examination, 27 miRNAs (15 upregulated and 12 downregulated) were found to be coassociated with the replication process of two viruses, while the expression levels of 15 and 5 miRNAs were significantly changed in CVA16- and EV71-infected cells, respectively. A great number of target genes of 27 common differentially expressed miRNAs were predicted by combined use of two computational target prediction algorithms, TargetScan and MiRanda. Comprehensive bioinformatic analysis of target genes in GO categories and KEGG pathways indicated the involvement of diverse biological functions and signaling pathways during viral infection. These results provide an overview of the roles of miRNAs in virus-host interaction, which will contribute to further understanding of HFMD pathological mechanisms.

Background Seven persons in one family living in eastern China developed fever and thrombocytopenia during May 2007, but the initial investigation failed to identify an infectious etiology. In December 2009, a novel bunyavirus (designated severe fever with thrombocytopenia syndrome bunyavirus [SFTSV]) was identified as the cause of illness in patients with similar clinical manifestations in China. We reexamined this family cluster for SFTSV infection. Methods We analyzed epidemiological and clinical data for the index patient and 6 secondary patients. We tested stored blood specimens from the 6 secondary patients using real time reverse transcription polymerase chain reaction (RT-PCR), viral culture, genetic sequencing, micro-neutralization assay (MNA), and indirect immunofluorescence assay (IFA). Results An 80-year-old woman with fever, leucopenia, and thrombocytopenia died on 27 April 2007. Between 3 and 7 May 2007, another 6 patients from her family were admitted to a local county hospital with fever and other similar symptoms. Serum specimens collected in 2007 from these 6 patients were positive for SFTS viral RNA through RT-PCR and for antibody to SFTSV through MNA and IFA. SFTSV was isolated from 1 preserved serum specimen. The only shared characteristic between secondary patients was personal contact with the index patient; none reported exposure to suspected animals or vectors. Conclusions Clinical and laboratory evidence confirmed that the patients of fever and thrombocytopenia occurring in a family cluster in eastern China in 2007 were caused by a newly recognized bunyavirus, SFTSV. Epidemiological investigation strongly suggests that infection of secondary patients was transmitted to family members by personal contact.

Usually, magnetic nanoparticles (MNPs) are prepared based on the famous Stober process in which divinylbenzene (DVA) is often used as a crosslink agent to synthesize SiO2/(PMMA/Fe3O4) nanoparticles. Compared with DVA, linolenic acid (LNA) is innoxious and can polymerize more easily for it has three unsaturated double bonds. In this paper, LNA was used as a new crosslink agent instead of DVA to synthesize the SiO2/(PMMA/Fe3O4) nanoparticles. The results showed that the core-shell structure could be observed obviously. The sizes of nanoparticles with core-shell structure range from 200 to 500 nm. The DNA probes which was immobilized on the surface of MNPs were used to capture the biotin modified complementary sequence of the probe, and the formed complexes were bonded with streptavidin-modified alkaline phosphatase (SA-AP). Finally the chemiluminescent signals were detected by adding 3-(2'-spiroadamantane)-4-methoxy-4-(3"-phosphoryloxy) phenyl-1, 2-dioxetane (AMPPD) which was the substrate reagent of AP. The specificity and sensitivity of this approach were investigated in this paper.

A dynamic computer simulation is carried out in the climates of 35 cities distributed around the world. The variation of the annual air-conditioning energy loads due to changes in the longwave emissivity and the solar reflectance of the building envelopes is studied to find the most appropriate exterior building finishes in various climates (including a tropical climate, a subtropical climate, a mountain plateau climate, a frigid-temperate climate and a temperate climate). Both the longwave emissivity and the solar reflectance are set from 0.1 to 0.9 with an interval of 0.1 in the simulation. The annual air-conditioning energy loads trends of each city are listed in a chart. The results show that both the longwave emissivity and the solar reflectance of building envelopes play significant roles in energy-saving for buildings. In tropical climates, the optical parameters of the building exterior surface affect the building energy-saving most significantly. In the mountain plateau climates and the subarctic climates, the impacts on energy-saving in buildings due to changes in the longwave emissivity and the solar reflectance are still considerable, but in the temperate continental climates and the temperate maritime climates, only limited effects are seen.

The system of chemiluminescent magnetic enzyme-linked immunoassay was developed. E. coli O157:H7 was sandwiched between rabbits anti-E. coli O157:H7 polyclonal antibody-coated magnetic nanoparticles (immunomagnetic nanoparticles or IMNPs) and mouse anti-E. coli O157:H7 monoclonal antibody. Commercial alkaline phosphatase conjugated horse anti-mouse immunoglobulin (ALP-Ab) was used to bind with the monoclonal antibody, finally the chemiluminescent signals were detected by adding 3-(2'-spiroadamantane)-4-methoxy-4-(3"-phosphoryloxy)phenyl-1,2-dioxetane (AMPPD) which was the substrate reagent of ALP. Different solvents of AMPPD were compared to get an optimal chemiluminescent signal. The effects of sodium borohydride and glycine on blocking the aldehyde groups of IMNPs were compared either, and the specificity and sensitivity of this system for detecting E. coli O157:H7 were researched. The results indicated that Tris buffer was the best solvent of AMPPD, sodium borohydride was better than glycine in blocking IMNPs, and this method was of good specificity when using E. coli Top 10F' and Vibrio cholera as negative controls. The detection limit was 10(3) cells mL(-1) when the antigen solution was 1 mL, and the procedure duration was about 3 h.

Highlights • A novel avian influenza A H7N9 virus has recently emerged in humans. • We present a duplex rRT-PCR assay for the low-cost detection of this virus. • The assay performed on a par with the WHO-recommended detection assay. • Assay sensitivity, specificity and reproducibility make it suitable for scaling-up., A novel reassortant influenza A (H7N9) virus emerged recently in China. In this study, a duplex real-time reverse transcription polymerase chain reaction (rRT-PCR) assay was developed for the simultaneous detection of hemagglutinin (HA) and neuraminidase (NA) genes of H7N9 influenza viruses. The sensitivity of the assay was determined to be 10 RNA copies per reaction for both HA and NA genes. No cross-reactivity was observed with other influenza virus subtypes or respiratory tract viruses. One hundred and forty-six clinical and environmental specimens were tested and compared with reference methods and were found to be consistent. The assay is suitable for large-scale screening due to short turnaround times and high specificity, sensitivity, and reproducibility.

Small submicroscopic DNA copy number variants represent an important source of variation in the human genome, human phenotypic diversity, and disease susceptibility. Consequently, there is a pressing need for the development of methods allowing the efficient, accurate, and cheap measurement of genomic copy number polymorphisms in clinical cohorts. The PCR-based strategies, being cost-effective and sensitive, are considered important in the development of screening techniques. PCR-based techniques such as multiplex PCR; multiplex ligation-dependent probe amplification; and a new single-tube assay technique, the competitive fluorescent multiplex STRP assay, have been applied to 22q11.2 detection, a typical example of deletion syndromes. In this study, we compared the reliability and application of these three techniques in a cohort of 17 patients affected with 22q11.2 deletion and 300 normal controls. All three techniques shared 100% sensitivity; however, the competitive fluorescent multiplex STRP assay had the lowest possibility of concurrent false-positive signals from two adjoining probes in a genomic region. Moreover, it is a relatively fast and low-cost procedure to detect the deletion of 22q11.2 in numerous patients with several minor symptoms of deletion syndromes. Multiplex PCR, a rapidly developing and cheap technique, allows detection of atypical deletions.

This paper described that Fe3O4 nanoparticles were encapsulated with methyl methacrylate (MMA) using linolenic acid (LA) as a crosslinking agent, after which the resulting polymethyl methacrylate (PMMA) embed Fe3O4 nanoparticles (PMMA/Fe3O4) were coated with silica, forming SiO2/ (PMMA/Fe3O4) core-shell structure particles. Then these magnetic nanoparticles (MNPs) were applied in the developed system of chemiluminescent magnetic enzyme-linked immunoassay. E. coli O157:H7 was sandwiched between rabbits anti-E. coli O157:H7 polyclonal antibody-coated magnetite nanoparticles (immunomagnetic nanoparticles or IMNPs) and mouse anti-E. coli O157:H7 monoclonal antibody (E. coli O157-McAb). Commercial alkaline phosphatase conjugated horse anti-mouse immunoglobulin (ALP-Ab) was used to bond with the monoclonal antibody, finally the chemiluminescent signals were detected by adding 3-(2'-spiroadamantane)-4-methoxy-4-(3"-phosphoryloxy)phenyl-1,2-dioxetane (AMPPD) which was the substrate reagent of ALP. The specificity and sensitivity of this system for detecting E. coli O157:H7 were researched. The results indicated that this method was of good specificity when using E. coli Top 10F' and Vibrio cholera as negative controls. The detection limit was 10(3) cells mL(-1) when the antigen solution was 1 mL, and the procedure duration was about 3 h.

SiO2/(PMMA/Fe3O4) composite particles were prepared from linolenic acid (LA) instead of oleic acid (OA) modified Fe3O4 nanoparticles by miniemulsion polymerization. LA has three unsaturated double bonds with which it can polymerizate more easily than OA. And coating Fe3O4 with polymethyl methacrylate (PMMA) polymer beforehand can prevent magnetic nanoparticles from the aggregation that usually comes from the increasing of ionic strength during the hydrolyzation of tetraethoxysilane (TEOS) by the steric hindrance. Finally, the resulting PMMA/Fe3O4 nanoparticles were coated with silica, forming SiO2/(PMMA/Fe3O4) core-shell structure particles. The sizes of nanoparticles with core-shell structure were in the range from 300 to 600 nm. The nanoparticles were spherical particles and had consistent size. The result of magnetic measurement showed that the composite particles had superparamagnetic property.

Matrix metallopeptidase-9 (MMP-9) plays a pivotal role in vascular remodeling and development of atherosclerotic lesion. The potentially functional MMP-9 polymorphisms may contribute to the susceptibility of coronary artery disease (CAD). A case–control study composed of 762 CAD cases and 555 CAD-free controls was conducted in a Chinese population to investigate the association between the MMP-9 −1562 C>T, R279Q, P574R and R668Q polymorphisms and CAD risk. It was found that the variant genotypes of R279Q, P574R and R668Q were associated with a non-significant decreased risk of CAD when compared with their wild-type genotypes, respectively, Furthermore, compared with those without any variant genotypes for these four nonsynonymouse loci, individuals carrying all four variant genotypes (−1562 CT/TT, 279 RQ/QQ, 574 PR/RR and 668 RQ/QQ) had a 51% decreased risk of CAD (adjusted OR = 0.49; 95% CI = 0.26–0.95, P = 0.033). Although no significant main effects were observed for MMP-9 −1562 C>T locus on CAD risk, variant genotypes of −1562 C>T were associated with a 2.53 increased risk of CAD in subjects with diabetes mellitus (DM) (95% CI = 1.18–5.45, P = 0.018). In CAD cases, variant genotypes of −1562 C>T were associated with a significantly increased risk of MI (adjusted OR, 1.48, 95% CI, 1.01–2.20, P = 0.048). These findings suggest that MMP-9 R279Q, P574R and R668Q may have combined effect in the occurrence of CAD and −1562 CT/TT genotypes may contribute to CAD in diabetics and MI in CAD patients.

Single nucleotide polymorphisms (SNPs) comprise the most abundant source of genetic variation in the human genome wide codominant SNPs identification. Therefore, large-scale codominant SNPs identification, especially for those associated with complex diseases, has induced the need for completely high-throughput and automated SNP genotyping method. Herein, we present an automated detection system of SNPs based on two kinds of functional magnetic nanoparticles (MNPs) and dual-color hybridization. The amido-modified MNPs (NH 2 -MNPs) modified with APTES were used for DNA extraction from whole blood directly by electrostatic reaction, and followed by PCR, was successfully performed. Furthermore, biotinylated PCR products were captured on the streptavidin-coated MNPs (SA–MNPs) and interrogated by hybridization with a pair of dual-color probes to determine SNP, then the genotype of each sample can be simultaneously identified by scanning the microarray printed with the denatured fluorescent probes. This system provided a rapid, sensitive and highly versatile automated procedure that will greatly facilitate the analysis of different known SNPs in human genome.

Summary Background In December, 2007, a family cluster of two individuals infected with highly pathogenic avian influenza A (H5N1) virus was identified in Jiangsu Province, China. Field and laboratory investigations were implemented immediately by public-health authorities. Methods Epidemiological, clinical, and virological data were collected and analysed. Respiratory specimens from the patients were tested by reverse transcriptase (RT) PCR and by viral culture for the presence of H5N1 virus. Contacts of cases were monitored for symptoms of illness for 10 days. Any contacts who became ill had respiratory specimens collected for H5N1 testing by RT PCR. Sera were obtained from contacts for H5N1 serological testing by microneutralisation and horse red-blood-cell haemagglutinin inhibition assays. Findings The 24-year-old index case died, and the second case, his 52-year-old father, survived after receiving early antiviral treatment and post-vaccination plasma from a participant in an H5N1 vaccine trial. The index case's only plausible exposure to H5N1 virus was a poultry market visit 6 days before the onset of illness. The second case had substantial unprotected close exposure to his ill son. 91 contacts with close exposure to one or both cases without adequate protective equipment provided consent for serological investigation. Of these individuals, 78 (86%) received oseltamivir chemoprophylaxis and two had mild illness. Both ill contacts tested negative for H5N1 by RT PCR. All 91 close contacts tested negative for H5N1 antibodies. H5N1 viruses isolated from the two cases were genetically identical except for one non-synonymous nucleotide substitution. Interpretation Limited, non-sustained person-to-person transmission of H5N1 virus probably occurred in this family cluster. Funding Chinese Ministry of Science and Technology; US National Institute of Allergy and Infectious Diseases, National Institutes of Health; China-US Collaborative Program on Emerging and Re-emerging Infectious Diseases.

Objective To explore the epidemiology and etiology for an outbreak of acute respiratory tract infection that occurred in one county of Jiangsu Province, China 2004. Methods Only cases meeting the case definition were included in the study. We reviewed the medical records of the cases who were admitted to the local hospitals, interviewed cases by a standard questionnaire, and then described the epidemiologic features and analyzed risk factors by means of a case-control study. We collected pharyngeal swab specimens and sent them to different laboratories for isolation and culture. The laboratory used different detection methods such as DIF, PCR, electron microscope examination and microneutralization assay, to identify and then type the positive specimens. Results A total of 871 cases were reported during the period from April 18 to July 4, 2004. The distribution of onset times presented two peaks, one in late May and another in middle June. The epidemic occurred mainly in the elementary and junior high schools in ten townships of one county, and the mean age of the cases was 12 years (range 7 months to 18 years). The course of the disease was acute, and was characterized by fever accompanied with sore throat and tonsillitis. The WBC count of cases was normal or elevated. The mean duration of illness was 5 days (range 2 to 12 days). No fatalities from illness were reported. A case-control study indicated that the possible risk factors were close contact with a case and/or poultry before onset and sharing of towels among members of the family. The typical CPE was observed through inoculating pharyngeal swab specimens into the HEP-2 cell cultures in different laboratories. An infection of adenovirus type 3 was verified by detecting positive specimens in different methods. Conclusion This investigation demonstrated that the acute respiratory infection in cases was caused by adenovirus type 3. Cases occurred in over 70 schools in ten townships in 2004, and the route of transmission was possibly close contact with cases or droplet transmission.

In April 2013, human infections with a novel avian influenza (H7N9) virus emerged in China. It has caused serious concerns for public health throughout the world. However, there is presently no effective treatment, and an A (H7N9) H7 subtype influenza vaccine is not available. Vaccination with virus-like particles (VLPs) has showed considerable promise for many other subtype influenza viruses. To produce H7N9 VLPs, full length, unmodified hemagglutinin (HA), neuraminidase (NA), and matrix1 (M1) genes from the A/Wuxi/1/2013(H7N9) were cloned into a pCDNA5.1 FRT vector. By co-transfection, VLPs containing HA, NA, and M1 were secreted by 293T cells. VLPs were purified by ultracentrifugation and injected into mice by the intramuscular route. In animal experiments, humoral and cellular immunoresponse were all triggered by H7N9 VLPs. High levels of specific antibodies and the isotypes of IgG were detected by ELISA. Anamnestic cellular immune responses were examined by detecting specific cytotoxic T cell for IFN-Υ production in ELISPOT assay. The hemagglutination-inhibition (HAI) against the homologous virus was more than 1:64, and cross-reactive HAI titers against the heterologous virus (H1N1 and H3N2) were more than 1:16. Moreover, VLPs immunized mice showed a rapid increase of neutralizing antibodies, with neutralizing antibody titers more than 1:8, which increased four-fold against PBS immunized mice in week four. By week six, the mice had high neutralization ability against the given strain and held a potent homologous virus neutralizing capacity. Thus, VLPs represent a potential strategy for the development of a safe and effective vaccine against novel avian influenza (H7N9) virus.

Background Sarcopenia is a common condition in older people. The aim of the present study was to examine the prevalence and factors associated with sarcopenia in an elderly Chinese suburb-dwelling population. Methods This study was conducted on 1,069 Chinese suburb-dwelling participants aged ≥60 years to evaluate sarcopenia using the Asian Working Group for Sarcopenia criteria. Sociodemographic and behavioral characteristics, as well as medical conditions, were considered independent variables to determine factors associated with sarcopenia using a logistic regression model. Results The prevalence of sarcopenia was 6.4% in men and 11.5% in women. Age was a significant factor in both sexes. In addition, presence of sarcopenia was inversely associated with BMI for both sexes. The odds ration and 95% confidence interval for factors statistically significantly associated with sarcopenia were 5.04 (1.70-14.89) and 2.36 (1.06-5.25) for diabetes in males and females, respectively; 10.60 (1.75-64.24) for daily consumption of alcohol (daily drinkers), 5.58 (2.13-14.59) for peptic ulcer in female (not statistically significant in males). Conclusions The Asian Working Group for Sarcopenia criterion is useful for defining sarcopenia, and our data suggest that the prevalence of sarcopenia in the general elderly suburb-dwelling Chinese population is high. Moreover, we find that high body mass index is inversely associated with the likelihood of being sarcopenic and that several others factors such as diabetes, peptic ulcer, and drinking habits increase the prevalence of sarcopenia.

Background: Circulating miRNAs are recently used as promising biomarkers for infectious diseases. The aim of this study was to identify plasma miRNAs for infection detection and monitoring the immune status of HIV infection. Methods: A cohort of 128 plasma samples from HIV-1 infected subjects and 37 samples from healthy donors were analyzed by TaqMan Low-Density Array (TLDA) method to find the differentially expressed miRNAs, in the light of HIV-1infected patients with low ( 350 cell/μL) CD4+ T cell count. Results: Seven miRNAs (miR-29a, miR-223, miR-27a, miR-19b, miR-151-3p, miR-28-5p, miR-766 and miR-30a-3p) were significantly associated with CD4+ T cell count (P Conclusions: We have identified highly sensitive and specific signatures of circulating miRNAs enabling non invasive detection and immune status monitoring of HIV-1 infection.

To establish a rapid eukaryotic expression system of hemagglutinin (HA) gene of novel avian influenza H7N9 using lentiviral vector, express the recombinant protein and study its functions in human embryonic kidney HEK293T cells.The full-length HA gene was amplified from H7N9 genomic RNA by reverse transcription PCR (RT-PCR) and linked with pMD18-T vector to generate pMD18-T-HA plasmid. Blunt-end HA gene with Kozak sequence was amplified from pMD18-T-HA vector, and then pLenti-HA-V5 expression vector was constructed by Topo cloning for transient expression in HEK293T cells. Expression of HA-V5 recombinant protein was confirmed by immunofluorescence assay (IFA) and Western blotting. Hemagglutination test was performed to evaluate the biological activity of the recombinant protein.The full-length HA gene (1 683 bp) was obtained and eukaryotic expression plasmid was constructed successfully. A recombinant protein with relative molecular mass (Mr) 70 000 was expressed and the antigenicity and binding specificity to positive serum were demonstrated by IFA and Western blotting. The hemagglutination activity was proved by hemagglutination test. IFA and Western blotting showed that the Mr 70 000 recombinant protein had an immuoreactivity to positive serum. The hemagglutination activity was confirmed by hemagglutination test.The rapid eukaryotic expression system of HA gene was successfully constructed, which laid a solid foundation for further research on subunit vaccine development, neutralizing epitope mapping and packaging pseudovirus.

Myeloid differentiation factor 88 (MyD88)-dependent IL-12 secretion by dendritic cells is critical for natural killer cell-mediated IFN-γ production and innate resistance to Toxoplasma gondii. Although MyD88(-/-) mice challenged with T. gondii have defective IL-12 responses and succumb to infection, administration of IL-12 to MyD88(-/-) mice fails to prevent acute mortality, suggesting that MyD88 may mediate signals within natural killer cells important for IL-12-dependent IFN-γ production and innate resistance to this parasite. In this study, we found that T. gondii antigens and IL-12 could synergistically trigger IFN-γ secretion by natural killer cells, which was dependent on toll-like receptor-MyD88 signaling. Further analysis showed that p38 mitogen-activated protein kinase, extracellular signal-regulated kinase, c-Jun N-terminal kinase and NF-κB multiple pathways downstream of MyD88 contributed to IFN-γ production by natural killer cells. Moreover, the well-established toll-like receptor agonists, T. gondii profilin (Tgprofilin) and T. gondii heat shock protein 70 (TgHSP70) could evoke a similar IFN-γ secretory response in natural killer cells to that evoked by T. gondii antigens. In vivo adoptive transfer experiments showed that, upon challenge with T. gondii, NOD/SCID-β2 microglobulin null (NOD/SCID-β2m(-/-)) mice injected i.v. with MyD88(-/-) natural killer cells had reduced serum IFN-γ levels and increased splenic tachyzoite burdens compared with those injected i.v. with wild-type natural killer cells. Taken together, these findings demonstrate a critical role for natural killer cell intrinsic toll-like receptor-MyD88 signaling in IL-12-dependent early IFN-γ production and innate resistance to T. gondii.