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Background Recurrent infections of animal hosts with avian influenza viruses (AIVs) have posted a persistent threat. It is very important to understand the avian influenza virus distribution and characteristics in environment associated with poultry and wild bird. The aim of this study was to analyze the geographic and seasonal distributions of AIVs in the 31 provinces, municipalities and autonomous region (PMA) of China, compare the AIVs prevalence in different collecting sites and sampling types, analyze the diversity of AIVs subtypes in environment. Methods A total of 742 005 environmental samples were collected from environmental samples related to poultry and wild birds in different locations in the mainland of China during 2014–2018. Viral RNA was extracted from the environmental samples. Real-time RT-PCR assays for influenza A, H5, H7 and H9 subtypes were performed on all the samples to identify subtypes of influenza virus. The nucleic acid of influenza A-positive samples were inoculated into embryonated chicken eggs for virus isolation. Whole-genome sequencing was then performed on Illumina platform. SPSS software was used to paired t test for the statistical analysis. ArcGIS was used for drawing map. Graphpad Prism was used to make graph. Results The nucleic acid positivity rate of influenza A, H5, H7 and H9 subtypes displayed the different characteristics of geographic distribution. The nucleic acid positivity rates of influenza A were particularly high (25.96%–45.51%) in eleven provinces covered the Central, Eastern, Southern, Southwest and Northwest of China. The nucleic acid positivity rates of H5 were significantly high (11.42%–13.79%) in two provinces and one municipality covered the Southwest and Central of China. The nucleic acid positivity rates of H7 were up to 4% in five provinces covered the Eastern and Central of China. The nucleic acid positivity rates of H9 were higher (13.07%–2.07%) in eleven PMA covered the Southern, Eastern, Central, Southwest and Northwest of China. The nucleic acid positivity rate of influenza A, H5, H7 and H9 showed the same seasonality. The highest nucleic acid positivity rates of influenza A, H5, H7, H9 subtypes were detected in December and January and lowest from May to September. Significant higher nucleic acid positivity rate of influenza A, H5, H7 and H9 were detected in samples collected from live poultry markets (LPM) (30.42%, 5.59%, 4.26%, 17.78%) and poultry slaughterhouses (22.96%, 4.2%, 2.08%, 12.63%). Environmental samples that were collected from sewage and chopping boards had significantly higher nucleic acid positivity rates for influenza A (36.58% and 33.1%), H5 (10.22% and 7.29%), H7(4.24% and 5.69%)and H9(21.62% and 18.75%). Multiple subtypes of AIVs including nine hemagglutinin (HA) and seven neuraminidase (NA) subtypes were isolated form the environmental samples. The H5, H7, and H9 subtypes accounted for the majority of AIVs in environment. Conclusions In this study, we found the avian influenza viruses characteristics of geographic distribution, seasonality, location, samples types, proved that multiple subtypes of AIVs continuously coexisted in the environment associated with poultry and wild bird, highlighted the need for environmental surveillance in China. Graphic Abstract

The aim of this paper is to understand the current situation of the care burden of patients with mental illness in remission and the factors affecting it in order to provide a scientific basis for targeted interventions. This paper reviews the concept of telemedicine, the application of telemedicine in home hospice care, and the remaining problems and improvement strategies of telemedicine in home hospice care, with the aim of providing a reference for the application of telemedicine in home hospice care in China. The Zarit Burden Scale, Family Care Scale, and Social Functioning Scale were used to conduct one-to-one interviews with 201 schizophrenic patients in remission and their primary caregivers in Hubei Province. Among them, 66, 72, and 25 cases (32.8%, 35.9%, and 12.4%) had mild, moderate, and severe burdens, respectively. Caregivers’ family care and patients’ social functioning were generally poor. The results of multiple linear regression analysis showed that caregiver age, caregiver education, caregiver family care, patient medical costs, and patient social functioning were factors influencing the burden of care for patients with schizophrenia in remission ( P < 0.05 ). The government, mental health centers, and families should understand the level of caregiving burden of patients with schizophrenia in remission and the factors influencing it and provide targeted measures to reduce the caregiving burden.

In mainland China, the disease burden of influenza is not yet fully understood. Based on population-based data, we aimed to estimate incidence rates of medically attended influenza and influenza virus infections in Ningbo City.We used data for outpatient acute respiratory illness (OARI) from a platform covering all health and medical institutes in Yingzhou District, Ningbo City. We applied generalized additive regression models to estimate influenza-associated excess incidence rate of OARI by age. We recruited local residents aged ≥60 years in the autumn of 2019 and conducted follow-up nearly 9 months later. Every survey, the sera were collected for testing hemagglutination inhibition antibody.From 2017-2018 to 2019-2020, the annual average of influenza-associated incidence rate of OARI in all ages was 10.9%. The influenza-associated incidence rate of OARI was the highest in 2017-2018 (16.9%) and the lowest in 2019-2020 (4.8%). Regularly, influenza-associated incidence rates of OARI were the highest in children aged 5-14 years (range: 44.1-77.6%) and 0-4 years (range: 8.3-46.6%). The annual average of excess OARI incidence rate in all ages was the highest for influenza B/Yamagata (3.9%). The overall incidence rate of influenza infections indicated by serology in elderly people was 21% during the winter season of 2019-2020.We identified substantial outpatient influenza burden in all ages in Ningbo. Our cohort study limited in elderly people found that this age group had a high risk of seasonal influenza infections. Our study informs the importance of increasing influenza vaccine coverage in high-risk population including elderly people.

Background: The first human-infected H9N2 influenza case can be traced back to 1998. Although the H9N2 influenza virus has low pathogenicity in animals, it donated partial or whole cassettes of internal genes to reassort novel viruses, such as H7N9, H10N8 and H5N6 viruses, that caused human infections with high fatality. Since 2013, sporadic but increasingly frequent human cases caused by H9N2 influenza virus have been confirmed globally, and most of them were from China. Methods: Information on human infections with H9N2 influenza virus was collected. Viral molecular determinants were determined by deep sequencing, and phylogenetic analysis was performed using MEGA 6.06. Antigenic analysis was performed by a hemagglutination inhibition (HI) assay. Receptor binding preference analysis was conducted based on a solid-phase binding assay with synthetic sialylglycopolymers. Antiviral susceptibility was determined by a fluorescence-based neuraminidase (NA) inhibition assay. Serological study of occupationally exposed populations was performed by HI assay screening and confirmed by microneutralization assay.Results: From 2013 to 2018, 33 human H9N2cases were reported in China, among them 75.7% were children under 10 years old .The 22 viruses were isolated and concentrated in the Y280/G9 lineage of the HA and NA genes. All human H9N2 viruses belonged to the Y280/G9 antigenic lineage, presented a human-like receptor binding preference and remained susceptible to NA inhibitors, but most demonstrated resistance to M2 inhibitors. The seroprevalence of occupationally exposed populations was 2.15%, 3.17%, 2.93% and 1.54% from 2015 to 2018, respectively. A significant difference in seroprevalence was shown between provinces with human cases (3.66%) and provinces without human cases (2.18%). Conclusions: The continuous antigenic drift and human-like receptor binding preference of the H9N2 virus enable it to have a high risk of causing human infections. The status of the seropositivity in occupationally exposed populations implies a substantial threat to public health. Research on human infection with H9N2 influenza virus should be strengthened to monitor the emergence of sustainable human-to-human transmission and the possibility of an endemic or a pandemic related to it.

Background Avian influenza viruses persist in animal hosts and continue to cause human infections in China. It is important to analyse the geographic and seasonal distributions of avian influenza viruses and compare the subtypes and their prevalence among sample sites in environment.MethodsA total of 329,276 environmental samples were collected from locations associated with poultry and wild birds from 2014 to 2017. Viral RNA was extracted from the environmental samples. Real-time PCR assays for influenza A, and the H5, H7, and H9 subtypes were performed on all the samples. Virus isolation was performed on the influenza A-positive samples detected by real-time PCR. Whole-genome sequencing was then performed on an Illumina sequencer.ResultsThe proportions of samples that tested positive for total influenza A and the H5, H9 and H7 subtypes varied among different geographical regions and seasons. Significantly higher proportions of influenza A- and H5-, H9-, and H7-positive samples were collected from live poultry markets and poultry slaughtering locations. Influenza A positivity rates in sewage and chopping board swab samples were higher than those in other sample types. Multiple subtypes related to avian influenza viruses, including 9 HA and 7 NA subtypes, were detected in environmental samples.ConclusionsThese findings indicate that multiple subtypes of avian influenza A viruses continuously coexist in environments associated with poultry and increase the risk of reassortment and transmission, highlighting the need for environmental surveillance in China.

The utilization of biomass and CO2 has become very important for solving environmental problems all over the world. In this study, a novel Fe-based CO2-gasification (FCG) process of biomass which combining chemical looping CO2 gasification and steam reforming process is proposed. The effects of experimental parameters including biomass/oxygen carrier (B/OC) ratio and initial CO2 concentration on the performance of the gas, liquid and solid products were analyzed in detail. The results show that CO2 has an opposite effect on the oxygen transfer capacity (OTC) of OC and carbon conversion rate, which results in an inflection point of initial CO2 concentration at around 20%: larger than this value, only a little hydrogen is generated, and once the concentration lower than this value, the yield of hydrogen dramatically increases. Moreover, the carbon conversion rate encountered a slight decline at the inflection point. As for the generation of tar, CO2 and OC have a synergetic effect on tar cracking: CO2 promotes the cracking of heavy tar and OC reduces the production of light tar effectively, and thus a better effect of tar cracking can be obtained when both OC and CO2 exist. This process can generate inherently separated syngas (CO-rich gas and high purity of H2), reduce the amount of undesired tar, utilize CO2 and Fe-containing solid waste, and thus this process is expected to have an extensive prospect for industrial application.

Background Since the highly pathogenic H5N1 influenza caused thousands of deaths of wild bird in this area in 2005, Qinghai Lake in China has become a hot spot for study of the influence of avian influenza to migratory wild birds. However, the ecology and evolution of low pathogenic avian influenza virus in this region are limited. This project-based avian influenza surveillance in Qinghai lake region was initiated in year 2012. Method Samples of wild bird feces and lake surface water were collected in Qinghai Lake in year 2012.Virus isolation was conducted on embryonated chicken eggs. The influenza A virus was determined by rRT-PCR. Virus sequences were acquired by deep sequencing. The phylogenetic correlation and molecular characteristics of the viruses were analyzed. The virus growth and infection features, receptor binding preference were studied, and pathogenicity in vitro as well as. Results Two H13N8 subtype influenza viruses were isolated. The viruses are phylogenetically belong to Eurasian lineage. Most of the genes are associated with gull origin influenza virus except PB1 gene, which is most probably derived from Anseriformes virus. The evidence of interspecies reassortment was presented. The two viruses have limited growth capacity on MDCK and A549 cells while grow well in embryonated eggs. The dual receptor binding features of the two viruses was shown up. The low pathogenic features were determined by trypsin dependence plaque formation assay. Conclusions The two H13N8 subtype influenza viruses are highly associated with gull origin. The interspecies reassortment of H13 subtype virus among Anseriforme sand Charadriiformes wild birds emphasizes the importance of strengthening avian influenza surveillance in this region. This study is helpful to understand the ecology, evolution and transmission pattern of H13 subtype influenza virus globally. Electronic supplementary material The online version of this article (10.1186/s12985-017-0842-1) contains supplementary material, which is available to authorized users.

As an eco-friendly transportation mode, shared bicycles provide a new option for public transit users in urban areas. China&rsquo, s bicycle-sharing market began flourishing in July 2016 and reached a plateau in 2017. How shared bicycles influenced public transit systems during this period is an interesting topic. A case study of Beijing is conducted. This study aims to identify the competitive and cooperative influences of shared bicycles on public transit by exploring the changes in public transit trip distances before and after the upsurge in bicycle-sharing. A histogram shifting method is introduced to examine the influences of shared bicycles on public transit services from a travel distance perspective. A spatial correlation of bicycling usage and public transit changes is calculated using units of gridded cell spaces. The results show: (1) overall transit usage continued growing after the shared bicycles market reached a plateau, (2) short public transits within 2 km decreased while transfers within 2 km increased, and (3) the decrease of short transits and increase of transfers within 3 km were spatially highly correlated to the usage of shared bicycles. Hence, the role of bicycle-sharing systems is competitive for existing public transit systems during short trips and cooperative for connecting transits.

H9N2 avian influenza viruses (AIVs) have become panzootic and caused sporadic human cases since 1998. Based on the poultry-related environmental surveillance data in mainland China from 2013 to 2016, a total of 68 representative environment isolates were selected and further investigated systematically. Phylogenetic analysis indicated that Y280-like H9N2 viruses have been predominant during 2013-2016 and acquired multiple specific amino acid substitutions that might favor viral transmission from avian to mammalians. Additionally, the viruses have undergone dramatic evolution and reassortment, resulting in an increased genetic diversity or acting as the gene contributors to new avian viruses. Receptor-binding tests indicated that most of the H9N2 isolates bound to human-type receptor, making them easily cross the species barrier and infect human efficiently. Our results suggested that the H9N2 AIVs prevalent in poultry may pose severe public health threat.

Experiments were conducted to investigate fluidization fundamentals of coarse particle using nitrogen as the fluidizing gas at high pressure. The particle under study was falling in Geldart’s group B/D. A 60-mm-dia. cylindrical Perspex model, a two-dimensional jetting fluidized bed, a 200-mm-semicircular jetting fluidized bed and a 200-mm-dia. cylindrical bed were used in the experiments. The fundamentals of high pressure fluidization examined in this study include minimum fluidization velocity and bed expansion. Empirical correlations were developed for determining minimum fluidization velocity and bed expansion. With the same excess fluidizing-gas velocity, bed expansion generally increases with the operating pressure, but the influence of pressure on bed expansion becomes weaker at higher pressure.

H6 avian influenza viruses (AIVs), which are prevalent in domestic and wild birds in Eurasian countries, have been isolated from pigs, a dog and a human. Routine virological surveillance at live poultry markets or poultry farms was conducted in southern China from 2009 to 2011. This study investigated the genetic and antigenic characteristics, analyzed the receptor-binding properties and evaluated the kinetics of infectivity of the AIVs in A549, MDCK and PK15 cells. A total of 14 H6N6 and 2 H6N2 isolates were obtained from four provinces in southern China. Genetic analysis indicated two distinct hemagglutinin lineages of the H6 strains cocirculating in southern China, and these strains facilitated active evolution and reassortment among multiple influenza virus subtypes from different avian species in nature. None of these isolates grouped with the novel Taiwan H6N1 virus responsible for human infection. Receptor-binding specificity assays showed that five H6 AIVs may have acquired the ability to recognize human receptors. Growth kinetics experiments showed that EV/HB-JZ/02/10(H6N2) and EV/JX/15/10(H6N6) initially reproduced faster and achieved higher titers than other viruses, suggesting that enhanced binding to α-2,6-linked sialic acids correlated with increased viral replication in mammalian cells. Overall, the results emphasize the need for continued surveillance of H6 outbreaks and extensive characterization of H6 isolates to better understand genetic changes and their implications.

A bubble-based EMMS model for a Pressurized Fluidized Bed (PFB) was revised on the basis of an energy minimization multi-scale (EMMS) method. In the new revised model, in order to calculate the heterogeneous index accounting for the hydrodynamic disparity between heterogeneous and homogeneous fluidization, a bubble size correlation derived from experiments on a pressurized jetting fluidized bed with a conical distributor was introduced. With given operating pressure and gas superficial velocity, a heterogeneous index was calculated by integrating the bubble size correlation under elevated pressure with the EMMS model. The heterogeneous index, which decreased with the height above the distributor and also had its lowest lever when the mean voidage is in the range of (emf, 1.0), was fitted into a surface and incorporated into Fluent as a user-defined function (UDF). The new bubble-based EMMS/PFB drag model was validated by predicting successfully the solid concentration distribution, the velocity of particles near the wall above the distributor, the radial profiles of particles' vertical motion and the average bubble size.

The highly pathogenic avian influenza (HPAI) H5N1 virus poses a significant potential threat to human society due to its wide spread and rapid evolution. In this study, we present a comprehensive antigenic map for HPAI H5N1 viruses including 218 newly sequenced isolates from diverse regions of mainland China, by computationally separating almost all HPAI H5N1 viruses into 15 major antigenic clusters (ACs) based on their hemagglutinin sequences. Phylogenetic analysis showed that 12 of these 15 ACs originated in China in a divergent pattern. Further analysis of the dissemination of HPAI H5N1 virus in China identified that the virus's geographic expansion was co-incident with a significant divergence in antigenicity. Moreover, this antigenic diversification leads to global antigenic complexity, as typified by the recent HPAI H5N1 spread, showing extensive co-circulation and local persistence. This analysis has highlighted the challenge in H5N1 prevention and control that requires different planning strategies even inside China.

Limited data are available regarding the swine influenza viruses (SIVs) that circulated in Mainland China prior to the 1990s. Eleven H3N2 virus strains were isolated from swine populations from 1979 to 1992. To determine the origin and tendency of these SIVs, the phylogenetic and antigenic properties of these viruses were analyzed based on the whole genome sequenced and the HI titrations with post-infection ferret antisera against influenza A (H3N2) virus isolates of swine and human origin. The results revealed that these 11 SIVs originated from humans and were not maintained in swine populations, indicating the interspecies transmission from humans to pigs occurred frequently and independently throughout these periods. However, human H3N2 viruses might not have the ability to circulate in pig herds.

In this paper, a facile near infrared (NIR, 650–900 nm) probe 4-dimethylamino-2′-hydroxychalcone (NIR-LP) based on the excited state intramolecular proton transfer (ESIPT) – aggregation induced emission (AIE) processes for the detection of the latent fingerprints (LFPs) was developed for the first time. The probe can distinguish the fresh LFPs, but also recognize the aged (10 days) LFPs.

Clade 2.3.4.4 highly pathogenic avian influenza viruses (H5Nx) have spread from Asia to other parts of the world. Since 2014, human infections with clade 2.3.4.4 highly pathogenic avian influenza H5N6 viruses have been continuously reported in China. To investigate the genesis of the virus, we analyzed 123 H5 or N6 environmental viruses sampled from live-poultry markets or farms from 2012 to 2015 in Mainland China. Our results indicated that clade 2.3.4.4 H5N2/N6/N8 viruses shared the same hemagglutinin gene as originated in early 2009. From 2012 to 2015, the genesis of highly pathogenic avian influenza H5N6 viruses occurred via two independent pathways. Three major reassortant H5N6 viruses (reassortants A, B, and C) were generated. Internal genes of reassortant A and B viruses and reassortant C viruses derived from clade 2.3.2.1c H5N1 and H9N2 viruses, respectively. Many mammalian adaption mutations and antigenic variations were detected among the three reassortant viruses. Considering their wide circulation and dynamic reassortment in poultry, we highly recommend close monitoring of the viruses in poultry and humans. IMPORTANCE Since 2014, clade 2.3.4.4 highly pathogenic avian influenza (H5Nx) viruses have caused many outbreaks in both wild and domestic birds globally. Severe human cases with novel H5N6 viruses in this group were also reported in China in 2014 and 2015. To investigate the genesis of the genetic diversity of these H5N6 viruses, we sequenced 123 H5 or N6 environmental viruses sampled from 2012 to 2015 in China. Sequence analysis indicated that three major reassortants of these H5N6 viruses had been generated by two independent evolutionary pathways. The H5N6 reassortant viruses had been detected in most provinces of southern China and neighboring countries. Considering the mammalian adaption mutations and antigenic variation detected, the spread of these viruses should be monitored carefully due to their pandemic potential.

The highly pathogenic avian influenza (HPAI) H5N1 virus poses a significant potential threat to human society due to its wide spread and rapid evolution. In this study, we present a comprehensive antigenic map for HPAI H5N1 viruses including 218 newly sequenced isolates from diverse regions of mainland China, by computationally separating almost all HPAI H5N1 viruses into 15 major antigenic clusters (ACs) based on their hemagglutinin sequences. Phylogenetic analysis showed that 12 of these 15 ACs originated in China in a divergent pattern. Further analysis of the dissemination of HPAI H5N1 virus in China identified that the virus’s geographic expansion was co-incident with a significant divergence in antigenicity. Moreover, this antigenic diversification leads to global antigenic complexity, as typified by the recent HPAI H5N1 spread, showing extensive co-circulation and local persistence. This analysis has highlighted the challenge in H5N1 prevention and control that requires different planning strategies even inside China.

Phylogenetic trees of a (PB2), b (PA), c (NP), d (M), and e (NS) of two H13N8 viruses. Full sequences were used to conduct phylogenetic tree using MEGA 7 with 1000 neighbor-joining replicates. Two H13N8 viruses isolated in Qinghai lake region are indicated by filled circles. (PDF 1 MB)

Summary Background Human infections with different avian influenza viruses—eg, H5N1, H9N2, and H7N9—have raised concerns about pandemic potential worldwide. We report the first human infection with a novel reassortant avian influenza A H10N8 virus. Methods We obtained and analysed clinical, epidemiological, and virological data from a patient from Nanchang City, China. Tracheal aspirate specimens were tested for influenza virus and other possible pathogens by RT-PCR, viral culture, and sequence analyses. A maximum likelihood phylogenetic tree was constructed. Findings A woman aged 73 years presented with fever and was admitted to hospital on Nov 30, 2013. She developed multiple organ failure and died 9 days after illness onset. A novel reassortant avian influenza A H10N8 virus was isolated from the tracheal aspirate specimen obtained from the patient 7 days after onset of illness. Sequence analyses revealed that all the genes of the virus were of avian origin, with six internal genes from avian influenza A H9N2 viruses. The aminoacid motif GlnSerGly at residues 226–228 of the haemagglutinin protein indicated avian-like receptor binding preference. A mixture of glutamic acid and lysine at residue 627 in PB2 protein—which is associated with mammalian adaptation—was detected in the original tracheal aspirate samples. The virus was sensitive to neuraminidase inhibitors. Sputum and blood cultures and deep sequencing analysis indicated no co-infection with bacteria or fungi. Epidemiological investigation established that the patient had visited a live poultry market 4 days before illness onset. Interpretation The novel reassortant H10N8 virus obtained is distinct from previously reported H10N8 viruses. The virus caused human infection and could have been associated with the death of a patient. Funding Emergency Research Project on human infection with avian influenza H7N9 virus, the National Basic Research Program of China, and the National Mega-projects for Infectious Diseases.

Objectives The first classical swine influenza A H1N1 viruses were isolated in Mainland China in 1991. To aid surveillance of swine influenza viruses as part of pandemic preparedness, we sought to identify their origin. Methods We sequenced and phylogenically analyzed 19 swine influenza viruses isolated in 1991 and 1992 in China and compared them with viruses isolated from other regions during the same period. Results All 19 swine influenza viruses analyzed in our study shared the highest similarity with the classical swine influenza virus A/Swine/Maryland/23239/1991 (H1N1). Phylogenetic trees of eight segmented genes exhibited similar topology, with all segments in the cluster of classical swine influenza viruses. In addition, antigenic analysis also indicated that the tested isolated were related to classical swine influenza isolates. Conclusions Classical swine H1N1 influenza viruses were predominant in Beijing pig herds during this period. Since both antibody and virus detections did not indicate the presence of CS H1N1 before 1991 in Mainland China, we combined with the data on pigs imported to and exported from China and concluded that these viruses might spread to China via pigs imported from North America and that they could affect the genetic evolution and transmission dynamics of swine influenza viruses in Hong Kong.

Human infection associated with a novel reassortant avian influenza H7N9 virus has recently been identified in China. A total of 132 confirmed cases and 39 deaths have been reported. Most patients presented with severe pneumonia and acute respiratory distress syndrome. Although the first epidemic has subsided, the presence of a natural reservoir and the disease severity highlight the need to evaluate its risk on human public health and to understand the possible pathogenesis mechanism. Here we show that the emerging H7N9 avian influenza virus poses a potentially high risk to humans. We discover that the H7N9 virus can bind to both avian-type (α2,3-linked sialic acid) and human-type (α2,6-linked sialic acid) receptors. It can invade epithelial cells in the human lower respiratory tract and type II pneumonocytes in alveoli, and replicated efficiently in ex vivo lung and trachea explant culture and several mammalian cell lines. In acute serum samples of H7N9-infected patients, increased levels of the chemokines and cytokines IP-10, MIG, MIP-1β, MCP-1, IL-6, IL-8 and IFN-α were detected. We note that the human population is naive to the H7N9 virus, and current seasonal vaccination could not provide protection.

Infection of poultry with influenza A subtype H7 viruses occurs worldwide, but the introduction of this subtype to humans in Asia has not been observed previously. In March 2013, three urban residents of Shanghai or Anhui, China, presented with rapidly progressing lower respiratory tract infections and were found to be infected with a novel reassortant avian-origin influenza A (H7N9) virus.We obtained and analyzed clinical, epidemiologic, and virologic data from these patients. Respiratory specimens were tested for influenza and other respiratory viruses by means of real-time reverse-transcriptase-polymerase-chain-reaction assays, viral culturing, and sequence analyses.A novel reassortant avian-origin influenza A (H7N9) virus was isolated from respiratory specimens obtained from all three patients and was identified as H7N9. Sequencing analyses revealed that all the genes from these three viruses were of avian origin, with six internal genes from avian influenza A (H9N2) viruses. Substitution Q226L (H3 numbering) at the 210-loop in the hemagglutinin (HA) gene was found in the A/Anhui/1/2013 and A/Shanghai/2/2013 virus but not in the A/Shanghai/1/2013 virus. A T160A mutation was identified at the 150-loop in the HA gene of all three viruses. A deletion of five amino acids in the neuraminidase (NA) stalk region was found in all three viruses. All three patients presented with fever, cough, and dyspnea. Two of the patients had a history of recent exposure to poultry. Chest radiography revealed diffuse opacities and consolidation. Complications included acute respiratory distress syndrome and multiorgan failure. All three patients died.Novel reassortant H7N9 viruses were associated with severe and fatal respiratory disease in three patients. (Funded by the National Basic Research Program of China and others.).

The human respiratory tract is a major site of avian influenza A(H5N1) infection. However, many humans infected with H5N1 present with gastrointestinal tract symptoms, suggesting that this may also be a target for the virus. In this study, we demonstrated that the human gut expresses abundant avian H5N1 receptors, is readily infected ex vivo by the H5N1 virus, and produces infectious viral particles in organ culture. An autopsy colonic sample from an H5N1-infected patient showed evidence of viral antigen expression in the gut epithelium. Our results provide the first evidence, to our knowledge, that H5N1 can directly target human gut tissues.

Background Avian influenza viruses have caused human infection and posed the pandemic potential. Live poultry markets are considered as a source of human infection with avian influenza viruses. Avian influenza routine surveillance of live poultry markets is taken annually in China. We isolated the 2 H11N9 influenza virus from the surveillance program. To better understand the risk caused by these new viruses, we characterize the genetic and pathogenicity of the two viruses. Methods Viral isolation was conducted with specific pathogen-free (SPF) embryonated chicken eggs. Whole genome was sequenced, and phylogenetic analysis was conducted. Results Two H11N9 viruses were identified, with all 8 segments belonging to the Eurasian lineage. The HA, NA, M, NS and PA genes were similar to virus isolates from ducks, and the NP, PB2 and PB1 gene segments were most similar to those viruses from wild birds, indicating that the H11N9 viruses might represent reassortant viruses from poultry and wild birds. The HA receptor binding preference was avian-like, and the cleavage site sequence of HA showed low pathogenic. The NA gene showed 94.6 % identity with the novel H7N9 virus that emerged in 2013. There was no drug resistance mutation in the M2 protein. The Asn30Asp and Thr215Ala substitutions in the M1 protein implied a potentially increased pathogenicity in mice. Both viruses were low-pathogenic strains, as assessed by the standards of intravenous pathogenicity index (IVPI) tests. Conclusion Two reassortant H11N9 avian influenza viruses were detected. These viruses showed low pathogenicity to chickens in the IVPI test. Public health concern caused by the reassortant H11N9 viruses should be emphasized during the future surveillance. Electronic supplementary material The online version of this article (doi:10.1186/s40249-016-0149-2) contains supplementary material, which is available to authorized users.

Indiana University-Purdue University Indianapolis (IUPUI)%%%%Pedestrian Automatic Emergency Braking (PAEB) system for avoiding/mitigating pedestrian crashes have been equipped on some passenger vehicles. At present, there are many e orts for the development of common standard for the performance evaluation of PAEB. The Transportation Active Safety Institute (TASI) at Indiana University-Purdue University-Indianapolis has been studying the problems and ad- dressing the concerns related to the establishment of such a standard with support from Toyota Collaborative Safety Research Center (CSRC). One of the important components in the PAEB evaluation is the development of standard testing facili- ties at night, in which 70% pedestrian crash social costs occurs [1]. The test facility should include representative low-illuminance environment to enable the examination of sensing and control functions of di erent PAEB systems. This thesis work focuses on modeling low-illuminance driving environment and describes an approach to recon- struct the lighting conditions. The goal of this research is to characterize and model light sources at a potential collision case at low-illuminance environment and deter- mine possible recreation of such environment for PAEB evaluation. This research is conducted in ve steps. The rst step is to identify lighting components that ap- pear frequently on a low-illuminance environment that a ect the performance of the PAEB. The identi ed lighting components include ambient light, same side/opposite side light poles, opposite side car headlight. Next step is to collect all potential pedes- trian collision cases at night with GPS coordinate information from TASI 110 CAR naturalistic driving study video database. Thirdly, since ambient lighting is relatively random and lack of a certain pattern, ambient light intensity for each potential col- lision case is de ned and processed as the average value of a region of interest on all video frames in this case. Fourth step is to classify interested light sources from the selected videos. The temporal pro le method, which compressing region of interest in video data (x,y,t) to image data (x,y), is introduced to scan certain prede ned region on the video. Due to the fact that light sources (except ambient light) impose distinct light patterns on the road, image patterns corresponding to speci c light sources can be recognized and classi ed. All light sources obtained are stamped with GPS coordinates and time information which are provided in corresponding data les along with the video. Lastly, by grouping all light source information of each repre- sentative street category, representative light description of each street category can be generated. Such light description can be used for lighting construction of PAEB test facility.

+T-cell immunity a b s t r a c t The development of pre-pandemic influenza A H5N1 vaccines that confer both antigen-sparing and crossclade protection are a high priority given the limited worldwide capacity for influenza vaccine production, and the antigenic and genetic heterogeneity of circulating H5N1 viruses. The inclusion of potent adjuvants in vaccine formulations may achieve both of these aims. Here we show that the addition of JVRS-100, an adjuvant consisting of cationic liposome–DNA complexes (CLDC) to a clade 1-derived H5N1 split vaccine induced significantly higher virus-specific antibody than unadjuvanted formulations, with a >30fold dose-sparing effect and induction of increased antigen-specific CD4+ T-cell responses in mice. All mice that received one dose of adjuvanted vaccine and subsequent H5N1 viral challenges exhibited mild illness, lower lung viral titers, undetectable spleen and brain viral titers, and 100% survival after either homologous clade 1 or heterologous clade 2 H5N1 viral challenges, whereas unadjuvanted vaccine recipients showed significantly increased weight loss, viral titers, and mortality. The protective immunity induced by JVRS-100 adjuvanted H5N1 vaccine was shown to last for over one year without significant waning. Thus, JVRS-100 adjuvanted H5N1 vaccine elicited enhanced humoral and T-cell responses, dosesparing, and cross-clade protection in mice. CLDC holds promise as an adjuvant for human pre-pandemic inactivated H5N1 vaccines.

Highly pathogenic influenza A (H5N1) virus causes a widespread poultry deaths worldwide. The first human H5N1 infected case was reported in Hong Kong Special Administrative Region of China in 1997. Since then, the virus re-emerged in 2003 and continues to infect people worldwide. Currently, over 400 human infections have been reported in more than 15 countries and mortality rate is greater than 60%. H5N1 viruses still pose a potential pandemic threat in the future because of the continuing global spread and evolution. Here, we summarize the epidemiological, clinical and virological characteristics of human H5N1 infection in China monitored and identified by our national surveillance systems.

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.

Human infection with avian influenza A H7N9 virus was first identified in March 2013 and represents an ongoing threat to public health. There is a need to optimize serological methods for this new influenza virus. Here, we compared the sensitivity and specificity of the hemagglutinin inhibition (HI), microneutralization (MN), and Western blot (WB) assays for the detection of human antibodies against avian influenza A (H7N9) virus. HI with horse erythrocytes (hRBCs) and a modified MN assay possessed greater sensitivity than turkey erythrocytes and the standard MN assay, respectively. Using these assays, 80% of tested sera from confirmed H7N9 cases developed detectable antibody to H7N9 after 21 days. To balance sensitivity and specificity, we found serum titers of ≥20 (MN) or 160 (HI) samples were most effective in determining seropositive to H7N9 virus. Single serum with HI titers of 20-80 or MN titer of 10 could be validated by each other or WB assay. Unlike serum collected from adult or elderly populations, the antibody response in children with mild disease was low or undetectable. These combinations of assays will be useful in case diagnosis and serologic investigation of human cases.

Background In the spring of 2013, a novel avian-origin influenza A (H7N9) virus in Eastern China emerged causing human infections. Concerns that a new influenza pandemic could occur were raised. The potential effect of chemical agents and physical conditions on inactivation of the novel avian influenza H7N9 virus had not been assessed. Methods To determine the inactivation effectiveness of the novel avian influenza A (H7N9) virus under various physical conditions and chemical treatments, two H7N9 viruses A/Anhui/1/2013 and A/Shanghai/1/2013 were treated by varied temperatures, ultraviolet light, varied pHs and different disinfectants. The viruses with107.7 EID50 were exposed to physical conditions (temperature, ultraviolet light and pH) or treated with commercial chemical agents (Sodium Hypochlorite, Virkon®-S, and Ethanol) respectively. After these treatments, the viruses were inoculated in SPF embryonated chicken eggs, the allantoic fluid was collected after 72–96 hours culture at 35°C and tested by haemagglutination assay. Results Both of the tested viruses could tolerate conditions under 56°C for 15 minutes or 60°C for 5 minutes, but their infectivity was completely lost under 56°C for 30 minutes, 65°C for 10 minutes, 70°C, 75°C and 100°C for 1 minute. It was also observed that the H7N9 viruses lost their infectivity totally after exposure of ultraviolet light irradiation for 30 minutes or longer time. Additionally, the viruses were completely inactivated at pH less than 2 for 0.5 hour or pH 3 for 24 hours, however, viruses remained infectious under pH treatment of 4–12 for 24 hours. The viruses were totally disinfected when treated with Sodium Hypochlorite, Virkon®-S and Ethanol at recommended concentrations after only 5 minutes. Conclusions The novel avian influenza A (H7N9) virus can be inactivated under some physical conditions or with chemical treatments, but they present high tolerance to moderately acidic or higher alkali conditions. The results provided the essential information for public health intervention of novel H7N9 avian influenza outbreak.

The outbreak of human infections caused by novel avian-origin influenza A(H7N9) in China since March 2013 underscores the need to better understand the pathogenicity and transmissibility of these viruses in mammals. In a ferret model, the pathogenicity of influenza A(H7N9) was found to be less than that of an influenza A(H5N1) strain but comparable to that of 2009 pandemic influenza A(H1N1), based on the clinical signs, mortality, virus dissemination, and results of histopathologic analyses. Influenza A(H7N9) could replicate in the upper and lower respiratory tract, the heart, the liver, and the olfactory bulb. It is worth noting that influenza A(H7N9) exhibited a low level of transmission between ferrets via respiratory droplets. There were 4 mutations in the virus isolated from the contact ferret: D678Y in the gene encoding PB2, R157K in the gene encoding hemagglutinin (H3 numbering), I109T in the gene encoding nucleoprotein, and T10I in the gene encoding neuraminidase. These data emphasized that avian-origin influenza A(H7N9) can be transmitted between mammals, highlighting its potential for human-to-human transmissibility.

Background Pigs are considered to be “mixing vessels” for the emergence of influenza viruses with pandemic potential. 2009 Pandemic Influenza H1N1 further proved this hypothesis, and raised the needs for risk assessment of human cases caused by swine influenza virus. Methods A field investigation was conducted after a case identified with infection of European avian-like swine influenza H1N1 virus. The diagnosis was confirmed by real-time PCR, virus isolation, whole genome sequencing and serological assays. Samples from local pigs and close contacts were tested to identify the source of infection and route of transmission. Results The virus from the index case was similar to viruses circulating in the local pigs. The case's grandfather was asymptomatic with sero-conversion. A total of 42.8% of swine sera were positive for European avian-like swine H1N1. Conclusions This study highlighted the importance of performing surveillance on swine influenza to monitor new virus emergence in humans.

To assess the extent of highly pathogenic avian influenza (HPAI) A (H5N1) virus transmission, we conducted sero-epidemiologic studies among close contacts exposed to H5N1 cases in mainland China during 2005–2008. Blood specimens were collected from 87 household members and 332 social contacts of 23 H5N1 index cases for HPAI H5N1 serological testing by modified horse red-blood-cell hemagglutinin inhibition and microneutralization assays. All participants were interviewed with a standardized questionnaire to collect information about the use of personal protective equipment, illness symptoms, exposure to an H5N1 case during the infectious period, and poultry exposures. Two (2.3%) household contacts tested positive for HPAI H5N1 virus antibody, and all social contacts tested negative. Both seropositive cases had prolonged, unprotected, close contact with a different H5N1 index case, including days of bed-care or sleeping together during the index case’s infectious period, and did not develop any illness. None of the 419 close contacts used appropriate personal protective equipment including 17% who reported providing bedside care or having physical contact with an H5N1 case for at least 12 hours. Our findings suggest that HPAI H5N1 viruses that circulated among poultry in mainland China from 2005–2008 were not easily transmitted to close contacts of H5N1 cases.

One of the primary efforts in influenza vaccine strain recommendation is to monitor through gene sequencing the viral surface protein haemagglutinin (HA) variants that lead to viral antigenic changes. Here we have developed a computational method, denoted as PREDAC, to predict antigenic clusters of influenza A (H3N2) viruses with high accuracy from viral HA sequences. Application of PREDAC to large-scale HA sequence data of H3N2 viruses isolated from diverse regions of Mainland China identified 17 antigenic clusters that have dominated for at least one season between 1968 and 2010. By tracking the dynamics of the dominant antigenic clusters, we not only find that dominant antigenic clusters change more frequently in China than in the United States/Europe, but also characterize the antigenic patterns of seasonal H3N2 viruses within China. Furthermore, we demonstrate that the coupling of large-scale HA sequencing with PREDAC can significantly improve vaccine strain recommendation for China.

Human infections of H5N1 highly pathogenic avian influenza virus have continued to occur in China without corresponding outbreaks in poultry, and there is little conclusive evidence of the source of these infections. Seeking to identify the source of the human infections, we sequenced 31 H5N1 viruses isolated from humans in China (2005 to 2010). We found a number of viral genotypes, not all of which have similar known avian virus counterparts. Guided by patient questionnaire data, we also obtained environmental samples from live poultry markets and dwellings frequented by six individuals prior to disease onset (2008 and 2009). H5N1 viruses were isolated from 4 of the 6 live poultry markets sampled. In each case, the genetic sequences of the environmental and corresponding human isolates were highly similar, demonstrating a link between human infection and live poultry markets. Therefore, infection control measures in live poultry markets are likely to reduce human H5N1 infection in China.