Construction of and evaluation of the immune response to two recombinant pseudorabies viruses expressing the B119L and EP364R proteins of African swine fever virus.

African swine fever (ASF) is an infectious disease caused by ASF virus (ASFV), which is characterized by high infectivity, rapid onset of disease, and a high mortality rate. Outbreaks of ASFV have caused great economic losses to the global pig industry, and there is a need to develop safe and effective vaccines. In this study, two recombinant pseudorabies virus (PRV) strains, rGXGG-2016-ΔgI/ΔgE-EP364R and rGXGG-2016-ΔgI/ΔgE-B119L, expressing the EP364R and B119L protein, respectively, of ASFV, were constructed by homologous recombination technology. Western blotting and immunofluorescence analysis showed that these foreign proteins were expressed in cells infected with the recombinant strains. The strains showed good genetic stability and proliferative characteristics for 20 passages in BHK-21 cells. Both of these strains were immunogenic in mice, inducing the production of specific antibodies against the expressed ASFV proteins while providing protection against lethal challenge with PRV. Thus, the recombinant strains rGXGG-2016-ΔgI/ΔgE-EP364R and rGXGG-2016-ΔgI/ΔgE-B119L could be used as candidate vaccines for both ASFV and PRV. In addition, our study identifies two potential target genes for the development of safe and efficient ASFV vaccines, provides a reference for the construction of bivalent ASFV and PRV vaccines, and demonstrates the feasibility of developing a live ASFV vector vaccine.

A S-Layer Protein of Bacillus anthracis as a Building Block for Functional Protein Arrays by In Vitro Self-Assembly.

S-layer proteins create a cell-surface layer architecture in both bacteria and archaea. Because S-layer proteins self-assemble into a native-like S-layer crystalline structure in vitro, they are attractive building blocks in nanotechnology. Here, the potential use of the S-layer protein EA1 from Bacillus anthracis in constructing a functional nanostructure is investigated, and apply this nanostructure in a proof-of-principle study for serological diagnosis of anthrax. EA1 is genetically fused with methyl parathion hydrolase (MPH), to degrade methyl parathion and provide a label for signal amplification. EA1 not only serves as a nanocarrier, but also as a specific antigen to capture anthrax-specific antibodies. As results, purified EA1-MPH forms a single layer of crystalline nanostructure through self-assembly. Our chimeric nanocatalyst greatly improves enzymatic stability of MPH. When applied to the detection of anthrax-specific antibodies in serum samples, the detection of our EA1-MPH nanostructure is nearly 300 times more sensitive than that of the unassembled complex. Together, it is shown that it is possible to build a functional and highly sensitive nanosensor based on S-layer protein. In conclusion, our present study should serve as a model for the development of other multifunctional nanomaterials using S-layer proteins.

[Assessment on analgesic and anti-inflammatory effects of Tongjingbao optimal formula and analysis on its active components].

OBJECTIVE: To assess the analgesic and anti-inflammatory effects of Tongjingbao optimal formula and analyze its active components. METHOD: Animals were divided into the model group, the Tongjingbao granule group and the Tongjingbao optimal formula group. The mice dysmenorrhea model was induced by oxytocin, and their content of blood calcium and MDA, NO, PGE2 in uterus were determined to assess the analgesic and anti-inflammatory effects of different components in Tongjingbao optimal formula and their impacts. RESULT: All components of Tongjingbao optimal formula could extend the dysmenorrhea incubation period of mice with dysmenorrhea, reduce their average writhing time, increase the writhing inhibition rate, lessen the content of blood calcium and MDA, PGE2 in uterus, and enhance the content of NO in uterus. CONCLUSION: All components of Tongjingbao optimal formula have the analgesic and anti-inflammatory effects, and different components show a synergistic effect in treating dysmenorrheal in many links.

[Effect of Vitamin D3 to Platelet Activation Mediated by Tumor Cell Culture Medium].

OBJECTIVE: To investigate the effect of vitamin D3 to platelet activation by tumor cell culture medium. METHODS: The peripheral blood platelets of BALB/c mice were isolated. The platelets were activated in 4T1 culture fluid for 24 h. The platelets were divided into 7 groups: control group, activation group, 1 nmol/L vitamin D3 group, 10 nmol/L vitamin D3 group, 50 nmol/L vitamin D3 group, 100 nmol/L vitamin D3 group, and positive drug (0.1 µmol/L eptifibatide) group. CCK-8 assay was used to detect the platelet proliferation at 24, 48 and 72 h. Flow cytometry was used to detect the expression of CD61 and CD62p and receptor for advanced glycation end products (RAGE) at 24, 48 and 72 h. ELISA was used to detect the level of platelet-endothelial cell adhesion molecule-1 (PECAM-1) at 24, 48 and 72 h. RESULTS: The CD41+/CD61+ and CD41+/CD62p+ ratio, PECAM-1 content and RAGE expression of platelets in activated group were all significantly increased as compared with those in control group (P<0.05). Compared with the activated group, the platelet proliferation, proportion of CD41+/CD61+ and CD41+/CD62p+, content of PECAM-1 and RAGE expression in vitamin D3 groups were all decreased (P<0.05). CONCLUSION: Vitamin D3 shows antiplatelet effect and can inhibit platelet proliferation and activation.

Percutaneous coronary intervention assisted by invasive mechanical ventilation and intra-aortic balloon pump for acute myocardial infarction with cardiogenic shock: Retrospective cohort study and meta-analyses.

There is little evidence to recommend the optimal invasive mechanical ventilation (IMV) modes and ideal positive end-expiratory pressure stress levels for acute myocardial infarction-cardiogenic shock (AMI-CS) patients. The aim of this study was to compare the mortality outcome in patients with AMI-CS who were treated with percutaneous coronary intervention (PCI) assisted by intra-aortic balloon pump (IABP) + IMV with historical controls. From January 1, 2016 to June 1, 2017, 60 patients were retrospectively enrolled at Tianjin Chest Hospital. Out of these, 88.3% of patients achieved thrombolysis in myocardial infarction (TIMI) flow 3 after PCI. The all-cause mortality rate in-hospital and at 1 year was 25% (95% CI: 0.14-0.36) and 33.9% (0.22-0.46), respectively. A systematic review followed by meta-analysis was performed with four historical studies of patients treated by PCI + IMV with partial IABP, which found an in-hospital mortality rate of 66.0% (95% CI: 0.62-0.71). Recently, a meta-analysis of patients receiving PCI + IABP with partial IMV showed that the 1 year mortality rate was 52.2% (95% CI: 0.47-0.58). In Cox regression analysis of patient data from the current study, lactic acid level ≥4.5 mmol/L, hyperuricemia, and TIMI flow <3 were independent predictors of death at 1 year. All-cause mortality, in-hospital and at 1 year, in patients with AMI-CS treated with PCI + IABP and IMV was lower than in those treated with PCI + partial IABP or IMV. Larger, longer-term direct comparisons are warranted.

[Scenario Analysis of PM2.5 Concentration Targets and Milestones in China].

Reports of decreasing PM2.5 concentrations in some developed countries and regions, as well as the trends of annual average concentrations of PM2.5 in the 74 key cities of China from 2013 to 2016 were analyzed. The cities were categorized based on PM2.5 concentration ranges and the regions where they are located. The average annual declining rates of PM2.5 concentration were calculated for these categories. Based on previous PM2.5 rates, we proposed different scenarios of decreasing PM2.5 concentration in Chinese cities for the future decades. Future PM2.5 concentration was calculated for each of the Chinese cities, and the milestones for 31 provinces and key areas were analyzed. The results showed that the annual average concentration of PM2.5 in China could meet the national air quality standard by 2025 and drop below 30 µg·m-3 in 2030 under both scenarios. The PM2.5 concentration in Beijing-Tianjin-Hebei and surrounding areas could meet the standards in 2030, and the Yangtze River Delta area in 2025. It will be difficult for Beijing, Tianjin, Hebei, and Henan to meet the standard in 2030. Even in the scenario where measures were intensified in the key areas, the cities failed to meet the PM2.5 concentration standards. In Beijing-Tianjin-Hebei and surrounding areas, the values were close to 40% of the target by 2030. To accelerate the reduction of PM2.5 concentration, extreme efforts will be needed in the highly polluted areas.

[PM2.5 Emission Characteristics and Estimation of Emission Reduction Potential from Typical Industrial Sources in Anyang].

To investigate the characteristics of PM2.5 emissions in Anyang, the PM2.5 emission inventory from industrial sources of Anyang in 2016 was established based on reasonable estimation method and emission factor. The spatial distribution analysis was carried out using Geographic Information System (GIS) technology. Based on the industry's typical ultra-low emission transformation and coal pressure reduction requirements, three scenarios were set to estimate the PM2.5 emission reduction potential of industrial sources from Anyang in 2020. The results show that the total PM2.5 emissions from industrial sources in Anyang was 81071.13 t in 2016. Non-ferrous metallurgy, iron and steel, and building materials industries were the main contributing sources, accounting for 45.43%, 25.74%, and 18.00% of the total emissions, respectively. The emission differences between towns and within Anyang were outstanding. The PM2.5 emissions were mainly concentrated in urban areas, Linzhou City, and Anyang County. The emissions in Anyang City were most prominent, while the emission intensity differences between the four jurisdictions of Anyang City were even greater. In the three different control scenarios, the potential for PM2.5 emission reduction in Anyang in 2020 was estimated to be 398.72, 11623.87, and 14072.27 t, accounting for 0.49%, 14.34%, and 17.22% of the total industrial source emissions in 2016, respectively. It can be concluded that Anyang has a great potential for PM2.5 emission reduction. In addition, ultra-low emission reform and coal pressure reduction can be of great significance for PM2.5 emission reduction in Anyang.

[Detection of HHV-8 Virus in 450 Free Blood Donons in Wuhan and Its Genotyping Based on ORFK1 Gene].

OBJECTIVE: To investigate the infection of HHV-8 virus in free blood donors in Wuhan. METHODS: Whole blood samples were collected from 450 free blood donors in Wuhan, and the genomic DNA extraction and serum collection were performed respectively. And then, the positive rate of HHV-8 was detected by PCR and ELISA, the positive detection rates were compared between them. Finally, HHV-8 ORFK1 gene was cloned by PCR in the positive samples, and the HHV-8 ORFK1 gene th was genetyped through sequencing analysis, homology comparison and phylogenetic tree construction. RESULTS: 25 and 23 cases of positive samples were detected by PCR and ELISA, their positive rate were 5.56% and 5.11% respectively, and without statistically significant difference using χ2 test analysis (P > 0.05). Based on the results of ORFK1 gene cloning and sequence analysis in 23 positive samples, 15 samples C subtype of had HHV-8 ORFK1 gene, and 8 cases had A subtype had HHV-8 ORFK1 gene. CONCLUSION: There is a certain percentage of HHV-8 infection in the free blood donors in Wuhan. It is suggested to increase the HHV-8 virus screening for free blood donors, so as to ensure the quality of blood.

Detection of Bacillus anthracis spores by super-paramagnetic lateral-flow immunoassays based on "Road Closure".

Detection of Bacillus anthracis in the field, whether as a natural infection or as a biothreat remains challenging. Here we have developed a new lateral-flow immunochromatographic assay (LFIA) for B. anthracis spore detection based on the fact that conjugates of B. anthracis spores and super-paramagnetic particles labeled with antibodies will block the pores of chromatographic strips and form retention lines on the strips, instead of the conventionally reported test lines and control lines in classic LFIA. As a result, this new LFIA can simultaneously realize optical, magnetic and naked-eye detection by analyzing signals from the retention lines. As few as 500-700 pure B. anthracis spores can be recognized with CV values less than 8.31% within 5 min of chromatography and a total time of 20 min. For powdery sample tests, this LFIA can endure interference from 25% (w/v) milk, 10% (w/v) baking soda and 10% (w/v) starch without any sample pre-treatment, and has a corresponding detection limit of 6×10(4) spores/g milk powder, 2×10(5) spores/g starch and 5×10(5) spores/g baking soda. Compared with existing methods, this new approach is very competitive in terms of sensitivity, specificity, cost and ease of operation. This proof-of-concept study can also be extended for detection of many other large-sized analytes.

[The impact of ambient particulate matter (PM10) on the population mortality for cerebrovascular diseases-a case-crossover study].

OBJECTIVE: To analyze the association between the concentration of ambient inhalable particulate matter (PM10) and population mortality for cerebrovascular diseases and to explore the impact of PM10 on cerebrovascular diseases. METHODS: Data including meteorological factors, air pollutants (NO2, SO2 and PM10) and cerebrovascular disease mortality in one district of Beijing from 2004 to 2008 were collected and both symmetric bidirectional case-crossover design and conditional logistic regression model were used to analyze the associations among them. RESULTS: After adjusting the influence of meteorological factors as daily average temperature and relative humidity, the single pollutant model showed that there was no significant lag effect. In the multi-pollutant model, the effect of the every 105.43 µg/m(3) increase of ambient PM10 had a larger impact on the daily death of the cerebrovascular diseases with statistically significant difference (P < 0.05). The effect of ambient PM10 pollution on daily death of cerebrovascular diseases was significant for females, 65 year-olds and in winter season. CONCLUSION: Our data showed that elevated levels of ambient PM10 was positively associated with the increase of cerebrovascular disease mortality. The elevated levels of ambient PM10 could lead to the increase of the daily mortality on cerebrovascular diseases for females, elderly who were 65 or older and in winter seasons.

Rapid detection of Bacillus anthracis spores using a super-paramagnetic lateral-flow immunological detection system.

There is an urgent need for convenient, sensitive, and specific methods to detect the spores of Bacillus anthracis, the causative agent of anthrax, because of the bioterrorism threat posed by this bacterium. In this study, we firstly develop a super-paramagnetic lateral-flow immunological detection system for B. anthracis spores. This system involves the use of a portable magnetic assay reader, super-paramagnetic iron oxide particles, lateral-flow strips and two different monoclonal antibodies directed against B. anthracis spores. This detection system specifically recognises as few as 400 pure B. anthracis spores in 30 min. This system has a linear range of 4×10³-106 CFU ml⁻¹ and reproducible detection limits of 200 spores mg⁻¹ milk powder and 130 spores mg⁻¹ soil for simulated samples. In addition, this approach shows no obvious cross-reaction with other related Bacillus spores, even at high concentrations, and has no significant dependence on the duration of the storage of the immunological strips. Therefore, this super-paramagnetic lateral-flow immunological detection system is a promising tool for the rapid and sensitive detection of Bacillus anthracis spores under field conditions.