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The organic soil is widely distributed around Dianchi Lake in Kunming, which is rich in the humic group (HG). In order to explore the effect of HG on the strength development of organic soil, this paper adopts the method of adding humic acid (HA) reagent into the undisturbed cohesive soil (blending method) and soaking it in fulvic acid (FA) solution (steeping method) to simulate the organic soil. Then, the effect of HG on soil is analyzed by the unconfined compressive strength (UCS) and the scanning electron microscope (SEM) tests. The UCS test shows that HA can significantly reduce the UCS of the samples, and it continues to decrease with the increase of the HA reagent content. FA can substantially enhance the UCS of the samples, which continues to increase with the concentration of FA solution (decreasing the pH value). Under each HG condition, the steeping of FA can significantly increase the UCS of the sample when the content of the HA reagent is less than 10%, and the growth rate of the UCS decreases when it is greater than 10%. With the increase of steeping age, the UCS of the 5% HA samples continued to increase and gradually became stable, and when the content of the HA reagent is more than 5%, the UCS curve first increases and then decreases. The SEM test shows that adding HA reagent can significantly increase the pore size, improve the connectivity between the pores, and weaken the connection of the soil structure. The steeping of FA can reduce the pore size, weaken the connectivity between the pores, and strengthen the connection of the sample structure. [ABSTRACT FROM AUTHOR]

The disposal of peat soil poses an increasingly difficult problem for actual engineering projects of Dianchi Lake area. This study obtains peat soil from seven areas around Dianchi Lake, and the content of humic acid (HA) in peat soil is between 2.36% and 28.13%. Then, this study simulates the peat soil by adding HA into the cohesive soil and uses cement to solidify it. The effect of cement and HA on the strength development of samples is examined by the unconfined compressive strength (UCS) test. Additionally, the microstructures of typical mixes are analyzed by scanning electron microscopy (SEM), X-ray diffraction (XRD), and mercury intrusion porosimetry (MIP). The results showed that HA will significantly reduce the UCS of cement soil. The SEM, XRD, and MIP confirmed that cement hydration reaction increases with cement ratio. In addition to the cementitious soil particles, hydration products gradually fill the pores of the soil and effectively reduce the number of large-size (6000∼40,000 nm) pores in cement soil, which makes the soil particle framework stronger. When the cement ratio increases from 15% to 25%, the diffraction peak of CAH and CSH increases faster. Combined with the results of the UCS test, it could be proved that cement ratio greater than 20% weakens the influence of HA on the strength development of cement soil sample. [ABSTRACT FROM AUTHOR]

To investigate the ice resistance of an icebreaker under the influence of snow cover, two series of tests, i.e. normal level ice and snow covered level ice, were performed with different navigating speeds in the Ice Engineering Laboratory of Tianjin University. The model ship was mounted on a rigid carriage through a one-directional load cell to measure the total ice resistance. A tactile sensor covering the area from stem to bow shoulder on one side of the model vessel was used in measuring the spatial distribution and temporal variation of the ice pressure on the bow area. Different ice cusp patterns and ice channel shapes were observed in different test series. The measurements indicate that the presence of snow cover increases the ice resistance both in the breaking and submersion components. The measured results are also compared with the calculated results through Lindqvist method, and some improvements and important discussions are made in the paper. [ABSTRACT FROM AUTHOR]

A standardized preparation process is proposed in this study for achieving optimal strength and vegetative properties in vegetated concrete, using Yunnan red soil as a growth substrate for plants. The porosity of vegetated concrete is a crucial factor influencing plant growth, while compressive strength is a significant mechanical property. To assess the strength and porosity of vegetated concrete, different design porosities (22%, 24%, 26%, 28%) and cement-to-aggregate ratios (4, 5, 6, 7) were utilized in the preparation of vegetated concrete samples. The shell-making and static-pressure-molding methods were optimized for specimen preparation. Analyzing the stress–strain full curve characteristics of vegetation-type concrete under different influencing factors, an in-depth investigation into its failure mechanism was conducted. It was determined that the design porosity and cement content significantly impact the concrete's performance, particularly in terms of 30-day compressive strength and effective porosity. Furthermore, an increase in the fly ash ratio led to an increase in porosity and a decrease in compressive strength, providing a certain guidance for optimizing concrete performance. Comparative analysis through vegetation experiments revealed that black rye grass exhibited favorable growth adaptability compared to other grass species. [ABSTRACT FROM AUTHOR]

Background: Diseases caused by Toxoplasma gondii (T. gondii) have introduced serious threats to public health. There is an urgent need to develop a rapid detection method for T. gondii infection in cats, which are definitive hosts. Recombinant apical membrane antigen 1 (rAMA1) was produced in a prokaryotic expression system and used as the detection antigen. The aim of this study was to evaluate and optimize a reliable indirect enzyme-linked immunosorbent assay (iELISA) method based on rAMA1 for the detection of antibodies against T. gondii in cats. Results: The rAMA1-iELISA method was developed and optimized by the chessboard titration method. There were no cross-reactions between T. gondii-positive cat serum and positive serum for other pathogens, indicating that rAMA1-iELISA could only detect T. gondii in most cases. The lowest detection limit of rAMA1-iELISA was 1:3200 (dilution of positive serum), and the CV of repeated tests within batches and between batches were confirmed to be less than 10%. The results of 247 cat serum samples detected by rAMA1-iELISA (kappa value = 0.622, p < 0.001) were in substantial agreement with commercial ELISA. The ROC curve analysis revealed the higher overall check accuracy of rAMA1-iELISA (sensitivity = 91.7%, specificity = 93.6%, AUC = 0.956, 95% CI 0.905 to 1.000) than GRA7-based iELISA (sensitivity = 91.7%, specificity = 85.5%, AUC = 0.936, 95% CI 0.892 to 0.980). Moreover, the positive rate of rAMA1-iELISA (6.5%, 16/247) was higher than that of GRA7-based iELISA (3.6%, 9/247) and that of commercial ELISA kit (4.9%, 12/247). Conclusion: The iELISA method with good specificity, sensitivity, and reproducibility was established and can be used for large-scale detection of T. gondii infection in clinical cat samples. [ABSTRACT FROM AUTHOR]

To study the corrosion behavior of cement soil in peat soil, the experiment involves the preparation of peat soil by incorporating humic acid into cohesive soil with a lower organic matter content. Cement soil samples are then prepared by adding cement to the mixture. These samples are subjected to immersion in fulvic acid solution and deionized water to simulate different working environments of cement soil. The experiment considers immersion time as the variable factor. It conducts observations of apparent phenomena, ion leaching tests, and unconfined compression strength tests on the cement soil. The experiment results are as follows: (1) With increasing immersion time, the surface of the cement soil in the peat soil environment experiences the disappearance of Ca(OH)2 and calcium aluminate hydrate. Additionally, large amounts of bird dropping crystals precipitate on the surface and within the specimen. The cement soil undergoes localized disintegration due to extensive erosion caused by swelling forces. (2) In the peat soil environment, fulvic acid reacts with the hydration products of cement, resulting in partial leaching of ions such as Ca2+, Mg2+, Al3+, and Fe3+ into the immersion solution. The lower the pH of the fulvic acid immersion (indicating higher concentration), the more significant the ion leaching. Increasing the ratio of humic acid to cement can slow down the leaching of ions. The cement soil undergoes dissolutive erosion in the peat soil environment. (3) The peat soil environment exerts both strengthening and corrosive effects on the cement soil. Cement soil without humic acid exhibits noticeable corrosion in the peat soil environment, gradually decreasing strength as immersion time increases. The strength decreases by 83% from 28 to 365 days. In contrast, cement soil with humic acid experiences an initial period of strengthening, leading to a significant increase in strength in the short term (34% increase from 28 to 90 days). However, the corrosive effects gradually dominate, resulting in a decrease in strength over time. The strength decreases by 80% from 90 to 365 days. This study also explores the strengthening effects of peat soil on cement soil. It identifies phenomena such as extensive erosion and new substance precipitation in cement soil. [ABSTRACT FROM AUTHOR]

Many peat soils are distributed around plateau lakes, and the reinforcement of peat soils with high organic matter content by ordinary cement cannot meet the actual engineering requirements. In order to obtain better mechanical properties and durability of the reinforcement, this experiment prepared peat soil by mixing humic acid reagent into the alluvial clay soil with low organic matter content. The cement soil samples were prepared by adding cement and ultrafine cement (UFC) by stirring method; the samples were then soaked in fulvic acid solution to simulate the cement soil in the peat soil environment. Using the unconfined compressive strength (UCS) test, scanning electron microscope (SEM) test, and pores and cracks analysis system (PCAS) test, the effect of UFC content change on cement soil's humic acid erosion resistance was explored, and the optimal UFC content range was sought. The results of the UCS test show that with an increase in immersion time, the strength curves of cement soil samples gradually increase to the peak strength and then decrease. Significant differences in the time correspond to the peak strength, and the overall presentation is two processes: the strength enhancement stage and the corrosion stage of the sample. The incorporation of UFC makes the cement soil in the peat soil environment exhibit excellent corrosion resistance, and the optimal UFC content is 10%. The results of the SEM and PCAS tests show that the microstructure of cement soil after immersion time exceeds 90 days, increases with an increase in immersion time, and its structural connectivity gradually weakens. The excellent characteristics of UFC particles, such as small particle size, narrow particle size distribution, fast hydration reaction rate, high hydration degree, and many hydration products, weakened the adverse effects of humic acid on the cement soil structure to a certain extent. Therefore, although the number of macropores increases, they are not connected. It still presents a relatively compact honeycomb overall structure, which correlates well with the UCS results. [ABSTRACT FROM AUTHOR]

Background: Although headache disorders are common, the current diagnostic approach is unsatisfactory. Previously, we designed a guideline-based clinical decision support system (CDSS 1.0) for diagnosing headache disorders. However, the system requires doctors to enter electronic information, which may limit widespread use. Methods: In this study, we developed the updated CDSS 2.0, which handles clinical information acquisition via human–computer conversations conducted on personal mobile devices in an outpatient setting. We tested CDSS 2.0 at headache clinics in 16 hospitals in 14 provinces of China. Results: Of the 653 patients recruited, 18.68% (122/652) were suspected by specialists to have secondary headaches. According to "red-flag" responses, all these participants were warned of potential secondary risks by CDSS 2.0. For the remaining 531 patients, we compared the diagnostic accuracy of assessments made using only electronic data firstly. In Comparison A, the system correctly recognized 115/129 (89.15%) cases of migraine without aura (MO), 32/32 (100%) cases of migraine with aura (MA), 10/10 (100%) cases of chronic migraine (CM), 77/95 (81.05%) cases of probable migraine (PM), 11/11 (100%) cases of infrequent episodic tension-type headache (iETTH), 36/45 (80.00%) cases of frequent episodic tension-type headache (fETTH), 23/25 (92.00%) cases of chronic tension-type headache (CTTH), 53/60 (88.33%) cases of probable tension-type headache (PTTH), 8/9 (88.89%) cases of cluster headache (CH), 5/5 (100%) cases of new daily persistent headache (NDPH), and 28/29 (96.55%) cases of medication overuse headache (MOH). In Comparison B, after combining outpatient medical records, the correct recognition rates of MO (76.03%), MA (96.15%), CM (90%), PM (75.29%), iETTH (88.89%), fETTH (72.73%), CTTH (95.65%), PTTH (79.66%), CH (77.78%), NDPH (80%), and MOH (84.85%) were still satisfactory. A patient satisfaction survey indicated that the conversational questionnaire was very well accepted, with high levels of satisfaction reported by 852 patients. Conclusions: The CDSS 2.0 achieved high diagnostic accuracy for most primary and some secondary headaches. Human–computer conversation data were well integrated into the diagnostic process, and the system was well accepted by patients. The follow-up process and doctor–client interactions will be future areas of research for the development of CDSS for headaches. [ABSTRACT FROM AUTHOR]

Background: Toxoplasma gondii is an obligate intracellular protozoan parasite that can cause a geographically widespread zoonosis. Our previous splenocyte microRNA profile analyses of pig infected with T. gondii revealed that the coordination of a large number of miRNAs regulates the host immune response during infection. However, the functions of other miRNAs involved in the immune regulation during T. gondii infection are not yet known. Methods: Clustering analysis was performed by K-means, self-organizing map (SOM), and hierarchical clustering to obtain miRNA groups with the similar expression patterns. Then, the target genes of the miRNA group in each subcluster were further analyzed for functional enrichment by Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and Reactome pathway to recognize the key signaling molecules and the regulatory signatures of the innate and adaptive immune responses of the host during T. gondii infection. Results: A total of 252 miRNAs were successfully divided into 22 subclusters by K-means clustering (designated as K1–K22), 29 subclusters by SOM clustering (designated as SOM1–SOM29), and six subclusters by hierarchical clustering (designated as H1–H6) based on their dynamic expression levels in the different infection stages. A total of 634, 660, and 477 GO terms, 15, 26, and 14 KEGG pathways, and 16, 15, and 7 Reactome pathways were significantly enriched by K-means, SOM, and hierarchical clustering, respectively. Of note, up to 22 miRNAs mainly showing downregulated expression at 50 days post-infection (dpi) were grouped into one subcluster (namely subcluster H3-K17-SOM1) through the three algorithms. Functional analysis revealed that a large group of immunomodulatory signaling molecules were controlled by the different miRNA groups to regulate multiple immune processes, for instance, IL-1-mediated cellular response and Th1/Th2 cell differentiation partly depending on Notch signaling transduction for subclusters K1 and K2, innate immune response involved in neutrophil degranulation and TLR4 cascade signaling for subcluster K15, B cell activation for subclusters SOM17, SOM1, and SOM25, leukocyte migration, and chemokine activity for subcluster SOM9, cytokine–cytokine receptor interaction for subcluster H2, and interleukin production, chemotaxis of immune cells, chemokine signaling pathway, and C-type lectin receptor signaling pathway for subcluster H3-K17-SOM1. Conclusions: Cluster analysis of splenocyte microRNAs in the pig revealed key regulatory properties of subcluster miRNA molecules and important features in the immune regulation induced by acute and chronic T. gondii infection. These results contribute new insight into the identification of physiological immune responses and maintenance of tolerance in pig spleen tissues during T. gondii infection. [ABSTRACT FROM AUTHOR]

Simple Summary: Toxoplasmosis is one of the most common parasitic zoonoses in the world. It not only threatens human health and safety but also causes considerable losses to the global animal husbandry industry. Swine, as an intermediate host of Toxoplasma gondii, has important economic and public health significance, and the global prevalence of T. gondii in swine varies between 10% and 60%. Therefore, it is important to establish simple, effective, sensitive and specific diagnostic methods for the detection and prevention of T. gondii infection in swine. In the present study, we developed a colloidal gold immunochromatographic strip based on rTgROP14, recombinant protein A and monoclonal antibody TgROP14-5D5 for serological detection of T. gondii in swine populations. This new ICT achieved good specificity and sensitivity and has potential for routine serological detection of T. gondii in swine. Toxoplasma gondii, a worldwide distributed apicomplexan protozoan, can infect almost all warm-blooded animals and may cause toxoplasmosis. In order to provide a point-of-care detection method for T. gondii infection, an immunochromatographic test (ICT) was established. The proposed test uses recombinant T. gondii rhoptry protein 14 (ROP14) conjugated with 20 nm gold particles, recombinant protein A as the detection line and monoclonal antibody TgROP14-5D5 as the control line. The specificity, sensitivity, positive predictive value, negative predictive value and stability of this new ICT were evaluated. rTgROP14 was specifically recognized by positive serum of T. gondii but not negative serum. mAb TgROP14-5D5 showed higher specific recognition of T. gondii antigens and was therefore selected for subsequent colloidal gold strip construction. The new ICT based on TgROP14 exhibited good diagnostic performance with high specificity (86.9%) and sensitivity (90.9%) using IHA as a "reference standard". Among 436 field porcine sera, ICT and IHA detected 134 (30.7%) and 99 (22.7%) positive samples, respectively. The relative agreement was 87.8%. These data indicate that this new ICT based on TgROP14 is a suitable candidate for routine testing of T. gondii in the field. [ABSTRACT FROM AUTHOR]

Background: Eimeria coccidiosis is a significant intestinal parasitic disease, which can lead to weight loss, disease and even death of many animals. At present, there is no information about the prevalence of Eimeria among the world's endangered species of Père David's deer (Elaphurus davidianus). Therefore, the purpose of this study is to identify an unknown Eimeria genus in the Père David's deer in Dafeng Milu National Nature Reserve, China. Results: A new Eimeria species is described from Père David's deer. Sporulated oocysts (n = 54) are pyriform, with a rough, yellowish brown, 2-layered oocyst wall (2.5 μm thick). A numerous small granules are dispersed randomly on the wall. Oocysts measured 41.2 (39.2–42.8) μm × 29.5 (27.9–30.5) μm, oocyst length/width (L/W) ratio, 1.4. Oocyst residuum, a polar granule and a polar cap are absent. The micropyle (3.5 μm wide) is present. Sporocysts are spindle shaped, 18.2 (16.5–20.0) μm × 10.5 (9.8–11.9) μm, sporocyst L/W ratio, 1.7 (1.5–1.9). A thin convex Stieda body is present and the sporocyst residuum is composed of numerous small granules less than 2.0 μm in diameter dispersed randomly. Each sporocyst contained 2 comma-shaped sporozoites in head-to-tail arrangement. A nucleus is located immediately anterior to the posterior, strong refractive and subspherical refractile body (~ 8 μm). Molecular analysis was conducted at the 18S, ITS-1 and COI loci. Conclusion: Based on the morphological and molecular data, this isolate is a new species of coccidian parasite, which is named Eimeria davidianusi after its host, the Père David's deer (Elaphurus davidianus). [ABSTRACT FROM AUTHOR]

Background: Toxoplasma gondii is an obligate intracellular protozoan parasite that can cause a geographically widespread zoonosis. Our previous splenocyte microRNA profile analyses of pig infected with T. gondii revealed that the coordination of a large number of miRNAs regulates the host immune response during infection. However, the functions of other miRNAs involved in the immune regulation during T. gondii infection are not yet known. Methods: Clustering analysis was performed by K-means, self-organizing map (SOM), and hierarchical clustering to obtain miRNA groups with the similar expression patterns. Then, the target genes of the miRNA group in each subcluster were further analyzed for functional enrichment by Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and Reactome pathway to recognize the key signaling molecules and the regulatory signatures of the innate and adaptive immune responses of the host during T. gondii infection. Results: A total of 252 miRNAs were successfully divided into 22 subclusters by K-means clustering (designated as K1–K22), 29 subclusters by SOM clustering (designated as SOM1–SOM29), and six subclusters by hierarchical clustering (designated as H1–H6) based on their dynamic expression levels in the different infection stages. A total of 634, 660, and 477 GO terms, 15, 26, and 14 KEGG pathways, and 16, 15, and 7 Reactome pathways were significantly enriched by K-means, SOM, and hierarchical clustering, respectively. Of note, up to 22 miRNAs mainly showing downregulated expression at 50 days post-infection (dpi) were grouped into one subcluster (namely subcluster H3-K17-SOM1) through the three algorithms. Functional analysis revealed that a large group of immunomodulatory signaling molecules were controlled by the different miRNA groups to regulate multiple immune processes, for instance, IL-1-mediated cellular response and Th1/Th2 cell differentiation partly depending on Notch signaling transduction for subclusters K1 and K2, innate immune response involved in neutrophil degranulation and TLR4 cascade signaling for subcluster K15, B cell activation for subclusters SOM17, SOM1, and SOM25, leukocyte migration, and chemokine activity for subcluster SOM9, cytokine–cytokine receptor interaction for subcluster H2, and interleukin production, chemotaxis of immune cells, chemokine signaling pathway, and C-type lectin receptor signaling pathway for subcluster H3-K17-SOM1. Conclusions: Cluster analysis of splenocyte microRNAs in the pig revealed key regulatory properties of subcluster miRNA molecules and important features in the immune regulation induced by acute and chronic T. gondii infection. These results contribute new insight into the identification of physiological immune responses and maintenance of tolerance in pig spleen tissues during T. gondii infection. [ABSTRACT FROM AUTHOR]

Wolbachia are maternally transmitted intracellular bacteria that can naturally and artificially infect arthropods and nematodes. Recently, they were applied to control the spread of mosquito-borne pathogens by causing cytoplasmic incompatibility (CI) between germ cells of females and males. The ability of Wolbachia to induce CI is based on the prevalence and polymorphism of Wolbachia in natural populations of mosquitoes. In this study, we screened the natural infection level and diversity of Wolbachia in field-collected mosquitoes from 25 provinces of China based on partial sequence of Wolbachia surface protein (wsp) gene and multilocus sequence typing (MLST). Among the samples, 2489 mosquitoes were captured from 24 provinces between July and September, 2014 and the remaining 1025 mosquitoes were collected month-by-month in Yangzhou, Jiangsu province between September 2013 and August 2014. Our results showed that the presence of Wolbachia was observed in mosquitoes of Aedes albopictus (97.1%, 331/341), Armigeres subalbatus (95.8%, 481/502), Culex pipiens (87.0%, 1525/1752), Cx. tritaeniorhynchus (17.1%, 14/82), but not Anopheles sinensis (n = 88). Phylogenetic analysis indicated that high polymorphism of wsp and MLST loci was observed in Ae. albopictus mosquitoes, while no or low polymorphisms were in Ar. subalbatus and Cx. pipiens mosquitoes. A total of 12 unique mutations of deduced amino acid were identified in the wsp sequences obtained in this study, including four mutations in Wolbachia supergroup A and eight mutations in supergroup B. This study revealed the prevalence and polymorphism of Wolbachia in mosquitoes in large-scale regions of China and will provide some useful information when performing Wolbachia-based mosquito biocontrol strategies in China. Author summary: The mosquitoes Aedes albopictus, Anopheles sinensis, Armigeres subalbatus, Culex pipiens and Cx. tritaeniorhynchus are native to China and the major vectors in the transmission of arboviruses, protozoans and nematodes. Recently, an innovative biocontrol strategy has been developed and evaluated based on the ability of Wolbachia to induce cytoplasmic incompatibility (CI), as well as interfere with the infection and replication of pathogens. Since the ability to induce CI largely depends on the density and diversity of Wolbachia, we investigated and characterized the natural infection of Wolbachia in above-mentioned five species of field-collected mosquitoes in 25 provinces of China. The results showed that the positive rates of Wolbachia infection were high in mosquitoes of Ae. albopictus, Ar. subalbatus and Cx. pipiens in large-scale regions of China and low in Cx. tritaeniorhynchus in Guizhou province. Phylogenetic analysis based on Wolbachia surface protein (wsp) gene and five multilocus sequence typing (MLST) loci indicated the high polymorphism of Wolbachia in Ae. albopictus, and low polymorphisms in Ar. subalbatus and Cx. pipiens. This finding contributes to the understanding of the nationwide distribution of Wolbachia and the potential application of this biocontrol strategy in China. [ABSTRACT FROM AUTHOR]

Background: Oxidation-specific epitopes (OSEs) are rich in atherosclerotic plaques. Innate and adaptive immune responses to OSEs play an important role in atherosclerosis. The purpose of this study was to develop novel human single-chain variable fragment (scFv) antibody specific to OSEs to image and inhibit atherosclerosis. Results: Here, we screened a novel scFv antibody, named as ASA6, from phage-displayed human scFv library. ASA6 can bind to oxidized LDL (Ox-LDL) and atherosclerotic plaques. Meanwhile, ASA6 can also inhibit the uptake of Ox-LDL into macrophage to reduce macrophage apoptosis. The atherosclerotic lesion area of ApoE−/− mice administrated with ASA6 antibody was significantly reduced. Transcriptome analysis reveals the anti-atherosclerosis effect of ASA6 is related to the regulation of fatty acid metabolism and inhibition of M1 macrophage polarization. Moreover, we conjugated ASA6 antibody to NaNdF4@NaGdF4 nanoparticles for noninvasive imaging of atherosclerotic plaques by magnetic resonance (MR) and near-infrared window II (NIR-II) imaging. Conclusions: Together, these data demonstrate the potential of ASA6 antibody in targeted therapy and noninvasive imaging for atherosclerosis. [ABSTRACT FROM AUTHOR]

Background: Toxoplasma gondii is an obligate parasite of all warm-blooded animals around the globe. Once infecting a cell, it manipulates the host’s DNA damage response that is yet to be elucidated. The objectives of the present study were three-fold: (i) to assess DNA damages in T. gondii-infected cells in vitro; (ii) to ascertain causes of DNA damage in T. gondii-infected cells; and (iii) to investigate activation of DNA damage responses during T. gondii infection. Methods: HeLa, Vero and HEK293 cells were infected with T. gondii at a multiplicity of infection (MOI) of 10:1. Infected cells were analyzed for a biomarker of DNA double-strand breaks (DSBs) γH2AX at 10 h, 20 h or 30 h post-infection using both western blot and immunofluorescence assay. Reactive oxygen species (ROS) levels were measured using 2′,7′-dichlorodihydrofluorescein diacetate (H2DCFDA), and ROS-induced DNA damage was inhibited by a ROS inhibitor N-acetylcysteine (NAC). Lastly, DNA damage responses were evaluated by detecting the active form of ataxia telangiectasia mutated/checkpoint kinase 2 (ATM/CHK2) by western blot. Results: γH2AX levels in the infected HeLa cells were significantly increased over time during T. gondii infection compared to uninfected cells. NAC treatment greatly reduced ROS and concomitantly diminished γH2AX in host cells. The phosphorylated ATM/CHK2 were elevated in T. gondii-infected cells. Conclusions: Toxoplasma gondii infection triggered DNA DSBs with ROS as a major player in host cells in vitro. It also activated DNA damage response pathway ATM/CHK2. Toxoplasma gondii manages to keep a balance between survival and apoptosis of its host cells for the benefit of its own survival. [ABSTRACT FROM AUTHOR]