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Alpine meadows are the largest grasslands in China. Greenhouse gas exchanges between alpine meadows and the atmosphere are highly uncertain due to the lack of year-round flux measurements. In this study, the methane (CH 4 ) and nitrous oxide (N 2 O) fluxes in an alpine Potentilla fruticosa shrub meadow in the Qinghai-Tibetan Plateau were investigated using the static, opaque chamber-gas chromatography method between April 2012 and April 2015. The annual CH 4 uptake (1.33–1.35 kg C ha −1 yr −1 ) and Q 10 value (1.79) were at the low end of the range for natural grasslands in China, which indicated that global warming at the same extent would result in less of an increase in the CH 4 sink in the Qinghai-Tibetan Plateau, compared to other grassland areas. The N 2 O emissions during the spring thaw period showed a tremendous inter-annual variation (0.03 to 0.14 kg N ha −1 ), which was closely linked to the variation in annual precipitation, especially the precipitation of the previous growing season. The high substrate concentrations and soil moisture during the spring thaw periods together provided the conditions for pulse N 2 O emissions. When the pulse N 2 O emissions occurred, emissions from the non-growing seasons dominated (67–74%) the annual total emissions. Thus, a proper sampling frequency (daily to weekly measurements) in non-growing seasons was needed for the quantification of annual fluxes. Four gas chromatographic set-ups (the Ar-CH 4 , N 2 -ascarite, N 2 -CO 2 , and pure N 2 methods) were adopted for N 2 O flux measurements over natural grasslands in China. Using the N 2 -CO 2 method, the annual N 2 O emissions from the alpine shrub meadow were quantified to be only 0.18–0.27 kg N ha −1 yr −1 in the present study. Based on measurements using the first three of the four gas chromatographic set-ups, the conclusion that can be drawn is that unfertilized natural grasslands in China function as marginally weak N 2 O sources, whereas the pure N 2 method may remarkably overestimate the emissions. [ABSTRACT FROM AUTHOR]

Rapid and high-throughput analytical techniques for IgE that requires a small serum amount are very important, especially for pediatric patients. In these patients, blood is collected from veins, which is painful compared to fingertip blood collection. Herein, a novel microfluidic system capable of high-throughput parallel analyses of allergen-specific IgE from small amounts of patient serum was successfully developed. A six-plex immunoassay was constructed within a microfluidic chip, and the entire system was validated using samples from clinical patients. Major antigens from house dust mite ( Dermatophagoides farinae and Blomia tropicalis ), cat ( Felis domesticus ), fungus ( Cladosporium herbarum ), ragweed ( Humulus japonicas ), and tree pollen ( Platanus acerifolia ) were used as analysis targets. Sample consumption decreased to <0.05 µL compared with the 480 µL serum consumption by fluoroenzyme immunoassay (UniCAP system Pharmacia Diagnostics AB, Uppsala, Sweden), the 50 µL serum consumption by enzyme-linked immune sorbent assay (ELISA), or the 1.5 µL serum consumption by conventional protein chip analysis. Analysis duration, reagent cost, and total cost for each measurement were also considerably decreased. The assay showed good accuracy and sensitivity toward the clinical samples. A significant correlation of allergen-specific IgE levels was found among the microfluidic assay, UniCAP system, and ELISA. [ABSTRACT FROM AUTHOR]

Abstract: Background: Rapid detection and identification of viruses are important for early diagnosis and effective surveillance of hand, foot, and mouth disease (HFMD). We described a novel assay using multilocus PCR and reverse transcription-PCR coupled with electrospray ionization mass spectrometry (RT-PCR/ESI-MS) to simultaneously detect and identify human enterovirus A–D, adenovirus A–F, human herpesvirus 1–8, parvovirus B19 and polyomavirus. Objectives: To evaluate the accuracy and efficacy of the RT-PCR/ESI-MS method, to detect and type enterovirus from specimens of clinical diagnosed HFMD patients. Study design: In this study, 152 specimens of clinically diagnosed HFMD patients were studied by the assay using RT-PCR/ESI-MS method. The detection and typing of enterovirus by RT-PCR/ESI-MS were compared with results from reference molecular methods. Results: The assay detected enteroviruses in 97 (63.8%) specimens, resulting in a sensitivity of 93.8% (95% CI: 91.8–95.7%) and a specificity of 87.5% (95% CI: 84.8–90.2%) compared with a reference clinical diagnostic test. Most enterovirus genotypes (65/84; 77%) determined by the platform were consistent with 5′ UTR sequence analysis, and most misidentifications resulted from the virus library, which could be further improved by updating the enterovirus database. In addition to enteroviruses, herpesviruses, polyomaviruses, adenoviruses and human rhinoviruses were detected and identified in 55 (36%) HFMD specimens by RT-PCR/ESI-MS. Conclusion: With the capability of high throughput and detection and typing of multiple clinically relevant viruses simultaneously, RT-PCR/ESI-MS can be a rapid and robust laboratory tool for identifying viral pathogens. [Copyright &y& Elsevier]

Apyrases (ATP diphosphohydrolase) hydrolyze the phosphodiester bonds of nucleoside tri- and diphosphates to orthophosphate and mononucleodides. They can inhibit platelet activation by depletion of adenosine diphosphate. In the current study, the Escherichia coli expression vector pET-19b equipped with an N-terminal histidine tag was applied to express the apyrase of Aedes albopictus. The gene-coding mature apyrase protein was amplified by RT-PCR and cloned into pET-19b. Soluble apyrase protein with high purity was successfully obtained by utilization of the suitable renaturation approach and Ni-NTA purification column. Four monoclonal antibodies to apyrase from A. albopictus were produced in male BALB/c mice immunized with the renatured apyrase. Using immunofluorescence assay and immunoblotting analysis, recombinant apyrase showed fine consistency with native apyrase. From kinetic analysis, it had a K of 11.6 μM and V of 1.02 nM/S/μg protein for adenosine triphosphate. Adenosine diphosphate-induced platelet aggregation was inhibited by approximately 6% when 0.4 μM recombinant apyrase was added and by about 9.5% when the concentration of recombinant apyrase was 0.8 μM. The effect on platelet aggregation was dose dependent. In conclusion, the apyrase of A. albopictus was cloned and expressed highly in the E. coli expression system. Recombinant apyrase protein showed biological activity, and anti-apyrase monoclonal antibody was also prepared. [ABSTRACT FROM AUTHOR]

• Diagnosis of three kinds of swine diseases for four different samples can be simultaneously achieved in a single chip. • The design of flow balance improves high consistency for multi-sample multi-biomarker testing. • The platform can realize reagent injection, well washing, waste collection and colorimetric detection automatically. • On-site testing for swine diseases achieves high accuracy, high sensitivity and high specificity. Early diagnosis and intervention of livestock epidemics is extremely important because disastrous outbreaks of an infectious disease can not only hit livestock industry but also cause a series of negative economic and social consequences. In order to fulfill the requirement of point-of-care testing, a reliable, field-portable, time-efficient and cost-effective microfluidic platform including a microfluidic chip and a semi-automated immunoassay system (SAIS) is developed. Multiple samples can be tested simultaneously in a single chip for multiple-biomarker testing. The entire assay, including reagent injection, well washing, waste collection and colorimetric detection, can be realized in the user-friendly system. The assay of three contagious diseases, PRRSV, CSFV and PCV2, is conducted in the microfluidic platform. The consistency of the reaction for different samples can be guaranteed owing to the chip design for flow balance. The assay only takes 12 min and 4 μL of sample is required. The assay achieves high accuracy (>93.8 %), high sensitivity (>88.8 %) and high specificity (>96.6 %). The platform has the potential to be applied for on-site rapid screening livestock diseases. [ABSTRACT FROM AUTHOR]

Background: Severe fever and thrombocytopenia bunyavirus (SFTSV) infection causes severe fever and thrombocytopenia syndrome with high mortality. It is extremely rare that a transmitting tick can be directly captured in bite wounds, and that SFTSV can be isolated from both the captured tick and patient's serum to establish a solid pathogen diagnosis. Case presentation: We report a case infected with severe fever and thrombocytopenia bunyavirus. The 69-year-old male patient presented with fever and tenderness on two lymph nodes in the right groin. A visible tick bite mark appeared on right upper quadrant of the patient's abdomen, and a live tick was captured in the bite wound upon physical examination. The virus was detected in both the blood of the patient and in the tick that stayed in the bite wound for 7 days. The phylogenetic analysis indicated that the SFTSV isolated from the tick and the patient's serum sample belonged to type B, in which the L/S segment of these two isolates shared 100% homology, while the M segment had 99.9% homology. The bitten patient was given various supportive care, but eventually died of multiple organ failure. Conclusion: The present case provides strong evidence of SFTSV transmission from H. longicornis to humans, and suggests that direct cross-species transmission can occur without additional intermediate hosts. [ABSTRACT FROM AUTHOR]

The intelligent security monitoring of buildings and their surroundings has become increasingly crucial as the number of high-rise buildings increases. Building structural health monitoring and early warning technology are key components of building safety, the implementation of which remains challenging, and the Internet of things approach provides a new technical measure for addressing this challenge. This article presents a novel integrated information system that combines Internet of things, building information management, early warning system, and cloud services. Specifically, the system involves an intelligent data box with enhanced connectivity and exchangeability for accessing and integrating the data obtained from distributed heterogeneous sensing devices. An extensible markup language (XML)–based uniform data parsing model is proposed to abstract the various message formats of heterogeneous devices to ensure data integration. The proposed Internet of things–based integrated information system structure was applied for monitoring an actual pit excavation engineering site. Three early warning levels were implemented according to rules based on the threshold value, which determined the specific safety personnel to be notified. The proposed Internet of things–based integrated information system is demonstrated to improve the effectiveness of monitoring processes and decision making in construction informatics applications. Our work highlights the crucial importance of a systematic approach toward integrated information systems for effective information collection and structural health monitoring. [ABSTRACT FROM AUTHOR]

The Internet of things (IoT) information system plays important roles in disposing of huge volumes of real-time service requests from heterogeneous devices, targeting for different complex application requirements. Load-dispatching control (LDC) is a key problem to be solved for devices accessing concurrently in cluster systems. Self-adaptive LDC optimizes the resource allocation to ensure no overloading node, thus, improving the performance of IoT systems. This paper focuses on adaptive dispatching control problem in IoT information system. First, a device data access platform is proposed for reducing the load imbalance and improving the efficiency of data processing. Then, we propose a processing capability prediction model to evaluate the system performance. On the basis of the model, we present a practical self-adaptive LDC framework with a self-adaptive control strategy and a load dispatching method. Finally, a case study is given to verify the framework and the control strategy. Experimental results show that the proposed strategy can meet the requirements of dynamic load balancing with the ability to avoid the load imbalance problem, and the LDC-based device access platform can process data accessing effectively and ubiquitously. [ABSTRACT FROM AUTHOR]

We report a novel method to analyze multiple cytokines/chemokines in 5 μL of BALF in mice. The methodology relies on micro-volume wall-less immunoassays combined with Luminex magnetic bead multiplex technology. This new method has been demonstrated with good validity, veracity, and sensitivity by comparing it with the existing conventional Luminex method, as well comparing the values of BALF cytokines/chemokines from functional inflammatory stimulated mice with that of healthy mice. Compared with conventional Luminex method, it also showed obvious advantages in some other aspects such as the sample volume was minimized to 5 μL, the consumption of reagents and beads was reduced by a factor of 5 and the total cost of experiments was significantly reduced by 80%, which would have wide applications in the field of multiple cytokines/chemokines analysis. [ABSTRACT FROM AUTHOR]

Toxoplasma gondii is a common opportunistic protozoan pathogen that can parasitize the karyocytes of humans and virtually all other warm-blooded animals. In the host's innate immune response to T. gondii infection, inflammasomes can mediate the maturation of pro-IL-1β and pro-IL-18, which further enhances the immune response. However, how intercellular parasites specifically provoke inflammasome activation remains unclear. In this study, we found that the T. gondii secretory protein, rhoptry protein 7 (ROP7), could interact with the NACHT domain of NLRP3 through liquid chromatography-mass spectrometry analysis and co-immunoprecipitation assays. When expressing ROP7 in differentiated THP-1 cells, there was significant up-regulation in NF-κB and continuous release of IL-1β. This process is pyroptosis-independent and leads to inflammasome hyperactivation through the IL-1β/NF-κB/NLRP3 feedback loop. The loss of ROP7 in tachyzoites did not affect parasite proliferation in host cells but did attenuate parasite-induced inflammatory activity. In conclusion, these findings unveil that a T. gondii-derived protein is able to promote inflammasome activation, and further study of ROP7 will deepen our understanding of host innate immunity to parasites. [ABSTRACT FROM AUTHOR]

In the chemical and biological fields, the creation of concentration gradient microenvironments is an important approach for many applications, such as crystal growth and drug screening. Although many concentration gradient generators have been demonstrated, current generators can hardly produce ultra-long linear concentration gradients. In this paper, we propose a concentration-gradient flow/droplet generator which consists of a microfluidic flow switch, a cavity array for stage-by-stage concentration dilution, and an optional T-junction for droplet formation. The generator can realize an ultra-long continuously-varying concentration gradient along the flow direction. Generation of a 38 mm concentration gradient was demonstrated. The length can be further extended by enlarging the capacity of the cavities and increasing the number of the stages. The concentration gradient showed high linearity in the range of 10% to 90%. Moreover, cyclic generation of a concentration gradient flow and droplets with different concentrations was realized by the generator. In a demonstration of drug screening, the generator was employed to produce paclitaxel in different concentrations. A negative correlation between the 4T1 cell viability and the paclitaxel concentration was observed after the treatment. We envision that the concentration gradient generator will be a promising candidate for various drug screening applications. [ABSTRACT FROM AUTHOR]

Based on empirical mode decomposition (EMD) and least squares support vector machine (LS-SVM), a new method is proposed to identify the flow pattern of gas–liquid two-phase flow in a minichannel. Four flow patterns are observed in three pipes with inner diameters of 4.0, 3.1, and 1.8 mm. For each flow pattern, the capacitance signals are obtained by a two-electrode capacitance sensor. The EMD method is applied to the capacitance signal to obtain intrinsic mode functions (IMFs) with different characteristic time scales. For each IMF, the autoregression (AR) model is built to extract multiscale features. Combining the extracted features with the energy feature of each IMF, the flow patterns are identified by the multiclassification LS-SVM classifier. The experimental results indicate that the presented method is effective for flow pattern identification and has identification rates higher than 91%. [ABSTRACT FROM PUBLISHER]

Microfluidics drives technological advancement in point-of-care (POC) bioanalytical diagnostics towards portability, fast response and low cost. In most microfluidic bioanalytical applications, flowing antigen/antibody reacts with immobilized antibody/antigen at a constant flux; it is difficult to reach a compromise to simultaneously realize sufficient time for the antigen–antibody interaction and short time for the entire assay. Here, we present a pump-free microfluidic chip, in which flow is self-initialized by capillary pumping and continued by imbibition of a filter paper. Microfluidic units in teardrop shape ensure that flow passes through the reaction areas at a reduced flux to facilitate the association between antigen and antibody and speeds up after the reaction areas. By spotting different antibodies into the reaction area, four types of biomarkers can be measured simultaneously in one microfluidic chip. Moreover, a small-sized instrument was developed for chemiluminescence detection and signal analysis. The system was validated by testing four biomarkers of colorectal cancer using plasma samples from patients. The assay took about 20 minutes. The limit of detection is 0.89 ng mL−1, 1.72 ng mL−1, 3.62 U mL−1 and 1.05 U mL−1 for the assays of carcinoembryonic antigen, alpha-fetoprotein, carbohydrate antigen 125 and carbohydrate antigen 19-9, respectively. This flux-adaptable and self-contained microfluidic platform is expected to be useful in various POC disease-monitoring applications. [ABSTRACT FROM AUTHOR]

Abstract A portion of alpine meadows has been and will continue to be cultivated due to the concurrent increasing demands for animal- and crop-oriented foods and global warming. However, it remains unclear how these long-term changes in land use will affect nitric oxide (NO) emission. At a field site with a calcareous soil on the Qinghai-Tibetan Plateau, the authors measured the year-round NO fluxes and related variables in a typically winter-grazed natural alpine meadow (NAM) and its adjacent forage oat field (FOF). The results showed that long-term plow tillage, fertilization and growing forage oats significantly yielded ca. 2.7 times more (p < 0.01) NO emissions from the FOF than the NAM (conservatively 208 vs. 56 g N/(ha·year) on average). The spring freeze–thaw period and non-growing season accounted for 17%‐35% of the annual emissions, respectively. The Q 10 of surface soil temperature (T s) was 8.9 in the NAM (vs. 3.8 in the FOF), indicating increases of 24%–93% in NO emissions per 1–3 °C increase. However, the warming-induced increases could be smaller than those due to land use change and management practices. The T s and concentrations of ammonium, nitrate and water-extractable organic carbon jointly explained 69% of the variance in daily NO fluxes from both fields during the annual period (p < 0.001). This result indicates that temporally and/or spatially distributed NO fluxes from landscapes with calcareous soils across native alpine meadows and/or fields cultivated with forage oats can be predicted by simultaneous observations of these four soil variables. Graphical abstract Unlabelled Image [ABSTRACT FROM AUTHOR]

Background: Entamoeba nuttalli is an intestinal protozoan with pathogenic potential that can cause amoebic liver abscess. It is highly prevalent in wild and captive macaques. Recently, cysts were detected in a caretaker of nonhuman primates in a zoo, indicating that E. nuttalli may be a zoonotic pathogen. Therefore, it is important to evaluate the pathogenicity of E. nuttalli in detail and in comparison with that of E. histolytica. Methodology/Principal findings: Trophozoites of E. nuttalli GY4 and E. histolytica SAW755 strains were inoculated into liver of hamsters. Expression levels of proinflammatory factors of hamsters and virulence factors from E. histolytica and E. nuttalli were compared between the two parasites. Inoculations with trophozoites of E. nuttalli resulted in an average necrotic area of 24% in liver tissue in 7 days, whereas this area produced by E. histolytica was nearly 50%. Along with the mild liver tissue damage induced by E. nuttalli, expression levels of proinflammatory factors (TNF-α, IL-6 and IL-1β) and amebic virulence protein genes (lectins, cysteine proteases and amoeba pores) in local tissues were lower with E. nuttalli in comparison with E. histolytica. In addition, M2 type macrophages were increased in E. nuttalli-induced amebic liver abscesses in the late stage of disease progression and lysate of E. nuttalli trophozoites induced higher arginase expression than E. histolytica in vitro. Conclusions/Significance: The results show that differential secretion of amebic virulence proteins during E. nuttalli infection triggered lower levels of secretion of various cytokines and had an impact on polarization of macrophages towards a M1/M2 balance. However, regardless of the degree of macrophage polarization, there is unambiguous evidence of an intense acute inflammatory reaction in liver of hamsters after infection by both Entamoeba species. [ABSTRACT FROM AUTHOR]

Background: Entamoeba histolytica is an intestinal protozoan parasite that causes amoebiasis, including amebic dysentery and liver abscesses. E. histolytica invades host tissues by adhering onto cells and phagocytosing them depending on the adaptation and expression of pathogenic factors, including Gal/GalNAc lectin. We have previously reported that E. histolytica possesses multiple CXXC sequence motifs, with the intermediate subunit of Gal/GalNAc lectin (i.e., Igl) as a key factor affecting the amoeba's pathogenicity. The present work showed the effect of immunization with recombinant Igl on amebic liver abscess formation and the corresponding immunological properties. Methodology/Principal Findings: A prokaryotic expression system was used to prepare the full-length Igl and the N-terminal, middle, and C-terminal fragments (C-Igl) of Igl. Vaccine efficacy was assessed by challenging hamsters with an intrahepatic injection of E. histolytica trophozoites. Hamsters intramuscularly immunized with full-length Igl and C-Igl were found to be 92% and 96% immune to liver abscess formation, respectively. Immune-response evaluation revealed that C-Igl can generate significant humoral immune responses, with high levels of antibodies in sera from immunized hamsters inhibiting 80% of trophozoites adherence to mammalian cells and inducing 80% more complement-mediated lysis of trophozoites compared with the control. C-Igl was further assessed for its cellular response by cytokine-gene qPCR analysis. The productions of IL-4 (8.4-fold) and IL-10 (2-fold) in the spleen cells of immunized hamsters were enhanced after in vitro stimulation. IL-4 expression was also supported by increased programmed cell death 1 ligand 1 gene. Conclusions/Significance: Immunobiochemical characterization strongly suggests the potential of recombinant Igl, especially the C-terminal fragment, as a vaccine candidate against amoebiasis. Moreover, protection through Th2-cell participation enabled effective humoral immunity against amebic liver abscesses. [ABSTRACT FROM AUTHOR]

Many colonies of macaques ( Macaca fascicularis and Macaca mulatta) are maintained in China, especially in Guangxi and Guizhou. A total of 803 fresh stool samples infected with Entamoeba were obtained from three big colonies of macaques located in southwest China. The samples were examined for the presence of five Entamoeba species using PCR. Entamoeba nuttalli, Entamoeba dispar, Entamoeba coli, and Entamoeba chattoni infections were detected, but Entamoeba histolytica infection was not. This study is the first to report on the prevalence of E. nuttalli in wild macaques from China. Eighteen E. nuttalli isolates and five E. dispar isolates were obtained by culturing the samples in Tanabe-Chiba medium. The serine-rich protein (SRP), ribosomal RNA (rRNA), hexokinase (HXK), glucose-6-phosphate isomerase (GPI), and phosphoglucomutase (PGM) genes of E. nuttalli isolates were compared with other reported isolates. The results showed clear differences among the Chinese E. nuttalli isolates and other isolates based on the SRP gene sequences. However, HXK, GPI, and PGM genes of these strains were similar to those of other isolates. The rRNA genes of E. coli and E. chattoni were also amplified and analyzed from these samples. The results suggested that host species might be a more important factor than geographic location in amebic genetic diversity. [ABSTRACT FROM AUTHOR]

A few PCR-based DNA typing methods using repetitive elements contained within both protein-coding genes and noncoding DNAs have been reported for Entamoeba histolytica over the years. The serine-rich E. histolytica protein and tRNA-linked short tandem repeats (STRs) are most commonly used to investigate the relationship between parasite genotype and E. histolytica infection outcome. Many E. histolytica infections in China have been reported; however, little genome information has been provided. In the current paper, five Chinese E. histolytica samples were reported: three amoebic liver abscess cases, one combined case and one asymptomatic case. Our study is the first to report on the DNA typing information of E. histolytica in China. We included two city, one imported, and two country cases. Sequence analysis of serine-rich protein genes confirmed the presence of seven sequence types in five isolates. The STRs amplified from the samples revealed five STR variations in the A-L, four in the N-K2, and R-R loci, three in D-A, S-D and S-Q loci. Two country patients were found to have a different outcome of infection with the same genotypes of E. histolytica, whereas in a city case, one E. histolytica strain had led to different outcome of the infection in one patient. Analyses of the results suggest that more genome information of E. histolytica strains from China through accurate methods is needed to interpret how the parasite genome plays a role in determining the outcome of E. histolytica infections. [ABSTRACT FROM AUTHOR]

Long-tailed macaques ( Macaca fascicularis) are bred in China for export and for use in experiments. Entamoeba infections in captive long-tailed macaques were surveyed in one of the biggest colonies located in Guangxi Province, China . One stool sample was obtained from each of the 152 different cages representing >3,000 macaques in the colony. The samples were examined by PCR for five Entamoeba species. The number of detected Entamoeba coli infections comprised 94% of the samples, 93% for Entamoeba chattoni, and 83% for Entamoeba dispar. In contrast, Entamoeba histolytica and Entamoeba nuttalli were not detected. Six isolates of E. dispar were obtained by culture in Tanabe-Chiba medium. Analysis of serine-rich protein genes in these isolates showed two genotypes, one of which is identical to that of the E. dispar SAW760 strain in humans. This suggests transmission of E. dispar between humans and nonhuman primates. These results demonstrate that Entamoeba infections are common, but virulent Entamoeba species are absent in this colony. This work also confirms the need for monitoring with PCR-based identification of Entamoeba species for captive macaques in breeding colonies to ensure animal health and protection of humans from zoonotic hazards. [ABSTRACT FROM AUTHOR]

The Yellow River basin is a key food production centre of global importance facing rapidly growing water scarcity. Water availability for agriculture in the basin is threatened by rapid growth in the demand for industrial and urban water, the need to flush sediment from the river's lower reaches, environmental demands and growing water pollution. Climate change is already evident in the basin with long-term declines in river runoff, higher temperatures, and increasing frequency and intensity of drought. The Chinese government has exhausted most options for improving water supply. The challenge will be to switch to improved water demand management, which is hampered by existing governance structures, and lack of integrated agriculture and water resource policies. [ABSTRACT FROM AUTHOR]

Air pollution has side effects on human health. Epidemiology studies indicate a positive association between ambient fine particle (PM 2.5 , or particles less than 2.5 μm in diameter) concentration and lung cancer. However, how fine particles affect lung cancer at the molecular level and related therapeutic methods to address these diseases are unclear. Here, the multi-omics analysis (DNA methylation and transcriptomic) was used to detect human bronchial epithelial cells (HBE), that were exposed to PM 2.5 using a quantified, small, portable, and organ-level air-liquid interface microfluidic system that mimics lung functions. The results indicate that 36,838 differentially methylated genes were detected. Of these 33,796 genes were hypomethylated (beta < 0), and 2862 genes were hypermethylated (beta > 0). RNA-Seq analysis demonstrated that 19,489 genes were upregulated (log 2 FC > 0), and 16,659 were downregulated. Furthermore, the calcium and apoptosis pathways were activated according to multi-omics analysis. The change in EGFR gene expression after PM 2.5 exposure was the result of alterations of the cellular DNA methylome in the promoter. Inhibition or down-regulation of EGFR could result in the regulation of the downstream intracellular Ca2+ concentration and apoptosis via the EGFR/PLCγ and EGFR/STAT/Bcl-XL pathways after PM 2.5 exposure. EGFR inhibitors decrease the Ca2+ concentration of cells, thereby strengthening the effects of fine particles on apoptosis. In short, the Ca2+ concentration and the apoptosis of cells can be regulated via EGFR related pathway after PM 2.5 exposure. The EGFR may be a potentially promising therapeutic target for the treatment of air pollution-induced lung cancer through regulation of the intracellular Ca2+ concentration and apoptosis. Unlabelled Image • A small air–liquid interface microfluidic system that mimics lung functions for cells exposure • EGFR gene expression change after PM 2.5 exposure was the result of the alteration of DNA methylome. • EGFR gene regulates calcium flux via the EGFR/PLCγ pathways after PM 2.5 exposure. • EGFR gene regulates apoptosis via the EGFR/STAT/Bcl-XL pathways after PM 2.5 exposure. [ABSTRACT FROM AUTHOR]

Rapid detection of severe fever with thrombocytopenia syndrome virus (SFTSV) is crucial for its control and surveillance. In this study, a rapid isothermal real-time reverse-transcription recombinase polymerase amplification (RT-RPA) assay was developed for the detection of SFTSV. The detection limit at 95% probability was 241 copies per reaction. A test of 120 serum samples of suspected severe fever with thrombocytopenia syndrome (SFTS) patients revealed that the sensitivity and specificity of the RT-RPA assay was approximately 96.00% (95%CI: 80.46%–99.79%) and 98.95% (95% CI: 94.28%–99.95%), respectively; the kappa value was 0.9495 (P＜0.001). The Bland-Altman analysis showed that 87.50% of the different data points were located within the 95% limits of agreement, indicating a good correlation between the results from RT-RPA assays and those of RT-qPCR assays. In conclusion, the rapid and efficient RT-RPA assay can be a promising candidate for point-of-care detection method of SFTSV. • A RT-RPA assay was developed to detect SFTSV RNA isothermally. • The assay can rapidly produce a result in 15 min at 39 °C. • The detection limit of the assay is 241 RNA sequences. • The results of RT-RPA compare well with RT-qPCR. • The RT-RPA assay may be used for field detection of SFTSV in resource-limited settings. [ABSTRACT FROM AUTHOR]

• A microfluidic chip and a semi-automated immunoassay instrument were demonstrated. • Reagent loading, well washing and signal detection can be procedurally realized. • Eleven samples can be simultaneously tested within a short time. • The platform was validated using clinical serum samples for human IL-6 ELISA. Microfluidic based enzyme-linked immunosorbent assay (ELISA) has been recognized as an advanced diagnosis tool due to the reduced sample consumption and fast reaction; but it often suffers from limited throughput and requires specialized operation skills. In this paper, we developed a microfluidic chip for multi-sample ELISA and an associated instrument for fluidic delivery and colorimetric detection. Eleven different samples can be assayed simultaneously using the developed platform in a straightforward, user-friendly, and low manual workload manner. Human interleukin-6 (IL-6) was used as the model protein biomarker to validate the platform and the estimated limit of detection (LOD) is 1.262 pg/ml. The entire assay took about 35 min, which is about six-fold shorter than the assay conducted in the traditional 96-well plate. Furthermore, we validated this platform using serum samples from 89 patients with ovarian cancer. The IL-6 concentrations in the clinical samples measured by the microfluidic platform were in good agreement with the traditional well-plate measurements. High accuracy, sensitivity and specificity of 95.5%, 96.2% and 90.9% are achieved in the testing of early-stage ovarian cancer. The semi-automated microfluidic immunoassay provides a convenient, rapid and reliable method for biomedical diagnosis. [ABSTRACT FROM AUTHOR]

Abstract: Acanthamoeba spp. can be parasitic in certain situations and are responsible for serious human infections, including Acanthamoeba keratitis, granulomatous amoebic encephalitis, and cutaneous acanthamoebiasis. We analyzed the fatty acid composition of Acanthamoeba castellanii trophozoites and tested the inhibitory activity of the main fatty acids, oleic acid and arachidonic acid, in vitro. Oleic acid markedly inhibited the growth of A. castellanii, with trophozoite viability of 57.4% at a concentration of 200 μM. Caspase‐3 staining and annexin V assays showed that apoptotic death occurred in A. castellanii trophozoites. Quantitative PCR and dot blot analysis showed increased levels of metacaspase and interleukin‐1β converting enzyme, which is also an indication of apoptosis. In contrast, arachidonic acid showed negligible inhibition of growth of A. castellanii trophozoites. Stimulated expression of Atg3, Atg8 and LC3A/B genes and monodansylcadaverine labeling suggested that oleic acid induces apoptosis by triggering autophagy of trophozoites. [ABSTRACT FROM AUTHOR]