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Objective This study investigates the effectiveness of tibia periosteum distraction (TPD) applied to the tibial periosteum, an innovative approach grounded in Ilizarov’s tension-stress theory, for the treatment of ischemic diabetic foot and vasculitic foot conditions. Methods A retrospective analysis was conducted on 33 patients (36 limbs) who underwent TPD between June 2019 and May 2022. The study comprised 21 males (23 limbs) and 12 females (13 limbs), aged 41 to 80 years (mean age: 63.4 years). Diabetic foot accounted for 27 cases, thromboangiitis obliterans for 2 cases, and arterial occlusive disease for 4 cases. The distribution of affected limbs included 15 left feet and 21 right feet. Periosteum traction commenced on the third postoperative day at a rate of approximately 0.75 mm/day, adjusted biweekly. The traction device was removed after two weeks. Evaluation included capillary refill and wound healing assessment, along with pre- and postoperative analysis of foot skin temperature, ankle-brachial index (ABI), visual analogue scale (VAS) pain scores, and peripheral blood oxygen saturation. CT angiography (CTA) was utilized to assess vascular conditions in both lower limbs. Results All 33 patients were successfully followed up for a duration ranging from 4 to 24 weeks (mean: 11.03 weeks). VAS pain scores significantly decreased from preoperative (5.09 ± 1.70, range: 2-8) to postoperative two weeks (2.24 ± 0.90, range: 1-4) (t = 9.44, p < .001). Oxygen saturation levels increased from 83.88% ± 11.82% (range: 58%-97%) preoperatively to 91.36% ± 5.69% (range: 76%-98%) at two weeks postoperatively (t = −4.21, p < .001). Foot skin temperature also showed a significant increase (t = −3.98, p < .001). Capillary refill test at two weeks postoperatively demonstrated notable improvement. CTA revealed evident neovascularization in the operated limbs compared to preoperative conditions. Wound improvement was significant in all 33 patients within two months postoperatively. Conclusion TPD emerges as a promising technique for chronic ischemic lower limb diseases, demonstrating favorable preliminary outcomes in wound healing promotion and amputation rate reduction. Nevertheless, large-scale randomized controlled trials are essential to further validate its efficacy.

Construction and Demolition Wastes (CDW) have a significant impact on global waste streams. Brick waste stands out as a prominent type of CDW, and numerous studies have explored its recycling for the creation of environmentally-friendly concrete. Reverse design of recycled brick aggregate concrete (RBAC) mixture proportion is presented in this paper with a focus on four key objectives, that is: compressive strength, cost, and environmental elements (i.e., energy consumption and carbon emission). Based on compiled experimental datasets of 374 samples, the back propagation neural network (BP), random forest (RF), and four meta-heuristic algorithm optimization models were constructed to achieve the desired compressive strength objective. In all machine learning (ML) methods, the compressive strength of RBAC can be predicted with high accuracy, with the SSA-BP (optimized back propagation neural network model using the sparrow search algorithm) model achieving superior results (i.e., NSE=0.91, RPD=3.2). The SSA-BP is therefore used as the objective function for compressive strength. The economic objective is primarily influenced by material costs, and the objective functions of energy consumption and carbon emission are determined by various aspects of production, transportation, and their mixing processes. In order to obtain the optimal RBAC design, the Non-Dominated Sorting Genetic Algorithm (NSGA-III) was implemented considering imperative constraints. Results indicate that cement amount and recycled brick aggregate (RBA)-to-natural aggregate proportion have a positive impact on the compressive strength. The suggested design framework allows for the creation of RBAC composite designs with varying levels of RBA substitution rates and strength targets, providing valuable guidance for tackling the CDW challenge and optimizing RBA usage.

Abstract Background Toxoplasma gondii is an obligate intracellular protozoan parasite that causes severe threats to humans and livestock. Macrophages are the cell type preferentially infected by T. gondii in vivo. Protein phosphorylation is an important posttranslational modification involved in diverse cellular functions. A rapidly accelerated fibrosarcoma kinase (A-Raf) is a member of the Raf family of serine/threonine protein kinases that is necessary for MAPK activation. Our previous research found that knockout of A-Raf could reduce T. gondii-induced apoptosis in porcine alveolar macrophages (3D4/21 cells). However, limited information is available on protein phosphorylation variations and the role of A-Raf in macrophages infected with T. gondii. Methods We used immobilized metal affinity chromatography (IMAC) in combination with liquid chromatography tandem mass spectrometry (LC–MS/MS) to profile changes in phosphorylation in T. gondii-infected 3D4/21 and 3D4/21-ΔAraf cells. Results A total of 1647 differentially expressed phosphorylated proteins (DEPPs) with 3876 differentially phosphorylated sites (DPSs) were identified in T. gondii-infected 3D4/21 cells (p3T group) when compared with uninfected 3D4/21 cells (pho3 group), and 959 DEPPs with 1540 DPSs were identified in the p3T group compared with infected 3D4/21-ΔAraf cells (p3KT group). Venn analysis revealed 552 DPSs corresponding to 406 DEPPs with the same phosphorylated sites when comparing p3T/pho3 versus p3T/p3KT, which were identified as DPSs and DEPPs that were directly or indirectly related to A-Raf. Conclusions Our results revealed distinct responses of macrophages to T. gondii infection and the potential roles of A-Raf in fighting infection via phosphorylation of crucial proteins. Graphical Abstract

Existing semi-empirical formulas for predicting punching shear capacity in FRP bar reinforced concrete flat slabs without shear reinforcement often prove inaccurate and unstable. This is primarily due to limited modeling data, inadequate consideration of key variables and neglect of complex nonlinear relationships. To address these challenges, this study delves into the utilization of advanced machine learning (ML) algorithms to offer precise and dependable estimates of punching shear capacity in such structural components. The study initially compiled a comprehensive database comprising 165 sets of test data, integrating eight crucial variables for model development. Subsequently, four data-driven models including back propagation artificial neural network (BPANN), support vector regression (SVR), random forest (RF) and gradient boosting regression tree (GBRT) were formulated to estimate the shear capacity. The efficacy of these models was assessed in comparison to existing prediction formulas. To interpret the models, this study also introduced shapley additive explanation (SHAP) and partial dependence plot (PDP) to quantitatively evaluate the influence of variables on predicted results. Research findings suggest that: (a) Among 25 existing formulas, Ju et al.’s approach performs notably well, with R2, Pre/Exp, MAPE and RMSE values at 0.76, 1.02, 22.2 % and 142.8 kN, respectively. (b) ML models surpass traditional formulas in predictive accuracy, with R2, Pre/Exp, MAPE and RMSE values ranging from 0.89 to 0.93, 1.03–1.09, 4.8–9.5 % and 55.4–69.0 kN, respectively. The GBRT model demonstrates the highest precision. (c) SHAP analysis of the GBRT model reveals that effective slab height and column section aspect ratio are pivotal variables influencing punching shear capacity. (d) PDP analysis quantitatively illustrates how punching shear capacity varies with each key variable.

Abstract Background The durable oocyst wall formed from the contents of wall-forming bodies (WFBs) protects Eimeria parasites from harsh conditions and enhances parasite transmission. Comprehending the contents of WFBs and proteins involved in oocyst wall formation is pivotal to understanding the mechanism of the oocyst wall formation and the search for novel targets to disrupt parasite transmission. Methods Total proteins extracted from WFBs and the oocyst wall of Eimeria necatrix were subjected to comparative proteomic analysis using tandem mass tag in conjunction with liquid chromatography tandem-mass spectrometry techniques. After functional clustering analysis of the identified proteins, three proteins, including E. necatrix disulfide isomerase (EnPDI), thioredoxin (EnTrx) and phosphoglycerate kinase (EnPGK), were selected for further study to confirm their potential roles in oocyst wall formation. Results A total of 3009 and 2973 proteins were identified from WFBs and the oocyst wall of E. necatrix, respectively. Among these proteins, 1102 were identified as differentially expressed proteins, of which 506 were upregulated and 596 downregulated in the oocyst wall compared to the WFBs. A total of 108 proteins, including compositional proteins of the oocyst wall, proteases, oxidoreductases, proteins involved in glycosylation, proteins involved in synthesis of the acid-fast lipid layer and proteins related to transport, were proposed to be involved in oocyst wall formation. The approximate molecular sizes of native EnPDI, EnTrx and EnPGK proteins were 55, 50 and 45 kDa, respectively. EnPDI was present in both type 1 and type 2 WFBs, EnTrx was present only in type 2 WFB2 and EnPGK was present only in type 1 WFBs, whereas all of them were localized to the outer layer of the oocyst wall, indicating that all of them participate in the formation of the oocyst wall. Conclusions To the best of our knowledge, this is the first report on the proteomes of WFBs and the oocyst wall of E. necatrix. The data obtained from this study form a basis for deciphering the molecular mechanisms underlying oocyst wall formation of Eimeria parasites. They also provide valuable resources for future studies on the development of novel therapeutic agents and vaccines aimed at combating coccidian transmission. Graphical Abstract

Abstract Background Toxoplasmosis is a zoonosis with a worldwide presence that is caused by the intracellular parasite Toxoplasma gondii. Active regulation of apoptosis is an important immune mechanism by which host cells resist the growth of T. gondii or avoid excessive pathological damage induced by this parasite. Previous studies found that upregulated expression of microRNA-185 (miR-185) during T. gondii infection has a potential role in regulating the expression of the ARAF gene, which is reported to be associated with cell proliferation and apoptosis. Methods The expression levels of miR-185 and the ARAF gene were evaluated by qPCR and Western blot, respectively, in mice tissues, porcine kidney epithelial cells (PK-15) and porcine alveolar macrophages (3D4/21) following infection with the T. gondii ToxoDB#9 and RH strains. The dual luciferase reporter assay was then used to verify the relationship between miR-185 and ARAF targets in PK-15 cells. PK-15 and 3D4/21 cell lines with stable knockout of the ARAF gene were established by CRISPR, and then the apoptosis rates of the cells following T. gondii infection were detected using cell flow cytometry assays. Simultaneously, the activities of cleaved caspase-3, as a key apoptosis executive protein, were detected by Western blot to evaluate the apoptosis levels of cells. Results Infection with both the T. gondii ToxoDB#9 and RH strains induced an increased expression of miR-185 and a decreased expression of ARAF in mice tissues, PK-15 and 3D4/21 cells. MiR-185 mimic transfections showed a significantly negative correlation in expression levels between miR-185 and the ARAF gene. The dual luciferase reporter assay confirmed that ARAF was a target of miR-185. Functional investigation revealed that T. gondii infection induced the apoptosis of PK-15 and 3D4/21 cells, which could be inhibited by ARAF knockout or overexpression of miR-185. The expression levels of cleaved caspase-3 protein were significantly lower in cells with ARAF knockout than in normal cells, which were consistent with the results of the cell flow cytometry assays. Conclusions Toxoplasma gondii infection could lead to the upregulation of miR-185 and the downregulation of ARAF, which was not related to the strain of T. gondii and the host cells. Toxoplasma gondii infection could regulate the apoptosis of host cells via the miR-185/ARAF axis, which represents an additional strategy used by T. gondii to counteract host-cell apoptosis in order to maintain survival and reproduce in the host cells. Graphical Abstract

IntroductionApicomplexan AP2 family of proteins (ApiAP2) are transcription factors (TFs) that regulate parasite growth and development, but little is known about the ApiAP2 TFs in Eimeria spp. ENH_00027130 sequence is predicted to encode a Eimeria necatrix ApiAP2 protein (EnApiAP2).MethodsThe cDNAs encoding full-length and truncated EnApiAP2 protein were cloned and sequenced, respectively. Then, the two cDNAs were cloned into the pET28a(+) expression vector and expressed expressed in Escherichia coli BL21. The mouse polyclonal antibody (pAb) and monoclonal antibody (mAb) against recombinant EnApiAP2 (rEnApiAP2) and EnApiAP2tr (rEnApiAP2tr) were prepared and used to localize the native EnApiAP2 protein in E. necatrix, respectively. Finally, the recombinant pEGFP-C1-ΔNLS-EnApiAP2s (knockout of a nuclear localization sequence, NLS) and pEGFP-C1-EnApiAP2 plasmid were constructed and transfected into DF-1 cells, respectively, to further observe subcellular localization of EnApiAP2 protein.ResultsThe EnApiAP2 gene had a size of 5019 bp and encoded 1672 amino acids, containing a conserved AP2 domain with a secondary structure consisting of an α-helix and three antiparallel β-strands. The rEnApiAP2 and rEnApiAP2tr were predominantly expressed in the form of inclusion bodies, and could be recognized by the 6×His tag mAb and the serum of convalescent chickens after infection with E. necatrix, respectively. The native EnApiAP2 protein was detected in sporozoites (SZ) and second generation merozoites (MZ-2) extracts, with a size of approximately 210 kDa. A quantitative real-time PCR (qPCR) analysis showed that the transcription level of EnApiAP2 was significantly higher in SZ than in MZ-2, third generation merozoites (MZ-3) and gametocytes (P

In order to explore the possible application of engineering waste soil (EWS) and recycled fine aggregate (RFA) in cement-based unfired clay bricks (CUCB), this paper utilizes the orthogonal experiment to investigate the combined utilization of EWS and RFA to fabricate eco-friendly CUCB. The study employs an L9 (34) orthogonal table with four factors and three levels for designing mix proportions. The factors include the ratios of water-to-cement (w/c), cement-to-EWS (c/e), RFA-to-EWS (r/e) and additive content. CUCB were tested for density, compressive strength, and flexural strength; and the impact of each factor on these properties was analyzed. The test results reveal that the eco-friendly CUCB exhibit lightweight characteristics and desirable mechanical properties. The ratio of flexural strength-to-compressive strength for eco-friendly CUCB ranges from 0.23 to 0.28, indicating that eco-friendly CUCB have good toughness. Data-driven models were developed to construct the relationships between target properties (i.e., density, compressive strength, and flexural strength) and four factors. The optimal mix proportion for physical and mechanical properties was determined to be w/c= 0.55, c/e = 0.3, r/e = 0.4 and additive content= 10%, with predicted compressive and flexural strengths of 19.68 MPa and 5.19 MPa, respectively. In addition, Scanning Electron Microscope (SEM) was performed to figure out the strength enhancement mechanism of eco-friendly CUCB with varying mix proportions. Environmental friendliness evaluation shows that the optimal mix pro-portion is more environmentally friendly to fabricate CUCB from the perspective of strength-normalized carbon footprint and energy consumption. Using large quantity of cement cannot increase the compressive strength and only acts as filler in the microstructure of eco-friendly CUCB.

The bamboo structure has received more and more attention as a green, renewable construction with an increasing focus on environmental issues. Cracking at joints or openings and brittle fracture of bamboo are urgent problems in bamboo structures, leading to reduced rigidity, decreased load capacity, and even endangering structural safety. This research aims to explore the mechanism of self-tapping screw reinforcement in improving the brittle fracture characteristic of bamboo laminate. To achieve an optimal effect of reinforcement, fracture tests of bamboo laminated with different numbers of self-tapping screws and screw positions were conducted to study their impact on fracture behaviors. Through the analysis of the ultimate load, the deformation capacity, the initial energy release rate, and the energy release rate caused by the self-tapping screws reinforcement, a model for calculating the energy release rate of bamboo laminate with self-tapping screws is established. Based on the model, the load-bearing capacity of steel-bamboo connections with self-tapping screws can be reasonably estimated.

The effects of process parameters on the microstructures and properties of the laser-assisted epitaxial growth layer are of great significance for the repair of nickel-based single-crystal blades. This work investigated the microstructures and properties of the epitaxial-growth laser-assisted nickel-based single-crystal superalloy. The effects of scanning path, laser power, scanning velocity and heat treatment on the microstructures and hardness of the laser-assisted epitaxial growth layer were explored. The microstructures of the epitaxial-growth samples are the same, starting from the bottom with a continuous columnar dendrite. The dendrites transform into isoaxial crystals when growing to a certain height. The scanning path, laser power, and scanning velocity mutually influence the sample's microstructures. The columnar crystal structure is very stable. Even the epitaxial-growth layer undergoes post-heat treatment, while the isoaxial crystal structure would be coarsened and the hardness decreases, but the subsequent aging treatment leads to an increase in hardness. These findings will greatly help improve the repair effect of single-crystal superalloys.

In present work, typical loading orientations were applied on an Al–Cu–Li single crystal during creep aging, and the relationship between precipitation behavior and loading orientations of the creep-aged Al–Cu–Li single crystal was studied by high angle annular dark field - scanning transmission electron microscope (HAADF-STEM) observation along multiple zone axes. The results show that the precipitation behavior of the precipitates (i.e., T1, θ′) in creep-aged Al–Cu–Li alloy was directly attributed to the loading directions. The number density of T1-phase in the creep-aged single crystal loaded close to α direction is highest, and that of T1-phase is the lowest when the loading direction is α. Moreover, the oriented-precipitation of T1-phase was found in the creep-aged sample loaded close to α direction, while that of θ′-phase was observed in the creep-aged sample loaded close to α direction. The hardness evolution of the creep-aged samples along different loading orientations is also in agreement with the corresponding precipitation behavior. The present work proposes a novel method to regulate the precipitation behavior of the creep-aged Al–Cu–Li alloy by controlling the loading direction, and the ideal loading direction is α for the Al–Cu–Li alloy to obtain better mechanical property.

Abstract Background Eimeria parasite infection occurs via ingestion of oocysts. The robust, bilayer oocyst wall is formed from the contents of wall-forming bodies (WFBs), WFB1 and WFB2, located exclusively in macrogametocytes. Eimeria necatrix gametocyte proteins 22 and 59 (EnGAM22 and EnGAM59) have been found to localize to WFBs and the oocyst wall. However, the exact localization of these two proteins is not clear. Methods WFBs of E. necatrix were extracted from purified gametocytes using a cutoff filter and the extracts of purified WFBs and gametocytes were analyzed using sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting. Then, the localization of EnGAM22 and EnGAM59 proteins was determined using an indirect immunofluorescence assay. Finally, the development of macrogametocytes and the oocyst wall of E. necatrix was analyzed using laser confocal microscopy and scanning electron microscopy. Results Purified WFBs had the same shape and size as those observed in macrogametocytes. EnGAM22 protein localized to WFB1, whereas EnGAM59 protein localized to WFB2. Both EnGAM22 and EnGAM59 native proteins were detected in the extracts of WFBs and gametocytes. The outer layer of the oocyst wall was formed by the release of the contents of WFB1 at the surface of the macrogametocyte to form an anti-EnGAM22 positive layer. WFB2 then appeared to give rise to the inner layer, which was anti-EnGAM59 positive. Conclusions EnGAM22 and EnGAM59 proteins localized to WFB1 and WFB2 and were involved in the formation of the outer and inner layers of the oocyst wall of E. necatrix, respectively. The processes of macrogametogenesis and oocyst wall formation of E. necatrix are similar to other Eimeria parasites. The anti-EnGAM22 antibody could be used as a tool to track the transport and secretion of proteins in WFB1 during oocyst development. Graphical Abstract

In this study, slow strain rate tensile tests under in situ hydrogen electrochemical charge are performed to investigate the hydrogen embrittlement (HE) behavior of selective laser-melted (SLM) and post-heat-treated Inconel 718 alloys in tandem with microstructure evolution and cracking feature characterization. The results indicate that the high density of dislocations and local strain along the cellular structure walls in the as-SLM sample increase local hydrogen concentration, leading to a crack along the cell walls of the cellular structure. The tensile strength and the HE susceptibility are significantly enhanced by the precipitation of the γ′, γ″, and δ phases generated by solution aging. Solution-aged samples suffer from the severe HE susceptibility caused by the δ phase formation along the grain boundaries and the cell walls, which lead to the δ/γ interface cracking.

Gastrointestinal (GI) parasites in small ruminants, especially goats and sheep, have caused significant socio-economic and public health challenges worldwide. The aim of the present study was to investigate the diversity and prevalence of GI parasites in goats and sheep in Jiangsu, Shaanxi and Hunan provinces of China, and to assess whether the age of animals, sampling season and feeding mode influence the distribution and infection of GI parasites. A total of 1,081 fecal samples collected from goats (n = 835) and sheep (n = 246) were detected by saturated saline flotation technique and nylon sifter elutriation and sieving method for eggs/oocysts, respectively. Based on the morphological observation of eggs and oocysts, one tapeworm, five nematodes, three trematodes and nineteen coccidia were identified, of which seven helminths belong to zoonotic parasites. The infection rate of parasites was 83.4% (902/1081) in total samples, 91.6% (765/835) in goats, and 55.7% (137/246) in sheep. The infection rate of coccidia was 71.0% (767/1081), and that of helminths was 56.2% (607/1081). The dominant species was E. alijeri (67.3%, 562/835) in goats, E. parva (30.1%, 74/246) in sheep. The highest prevalent helminths were Trichostrongylidae spp. in goats (58.3%, 487/835), and Moniezia spp. in sheep (22.76%, 56/246). Of 902 positive samples, 825 (91.5%, 825/902) contained multiple (2–10) parasites. The feeding mode, sampling season and regions were relevant risk factors which have significant influence on the occurrence of GI parasites in goats and sheep. The risk coefficient of parasite infection in autumn was 2.49 times higher than spring (Odds ratio = 2.49, 95% CI = 1.51–4.09, p

In recent years crushing waste brick to produce recycled brick aggregates (RBAs) has become a viable solution for reducing environmental pollution and addressing the natural resource shortage in civil engineering. To promote the widespread use of the recycled brick aggregate concrete (RBAC) in construction, this study analyzes existing test results on the attributes of RBAs and the compressive mechanical behaviors of RBAC. The review results indicate significant differences and variabilities in the characteristics of RBAs compared to natural coarse aggregates and recycled concrete coarse aggregates. RBAs have the highest absorption capacity and crushing index among the three aggregates, leading to changes in the compressive failure mechanism and a decline in the mechanical properties of RBAC. Additionally, it is also observed that existing formulas do not adequately account for the deterioration of the compressive mechanical properties of RBAC. To tackle this problem, artificial intelligence (AI) approaches including artificial neural network and multigene genetic programming are utilized to develop precise models for predicting the compressive strength and elastic modulus of RBAC. It is found that RBAC’s these two mechanical indexes are mainly influenced by the standard strength of cement paste, water-to-cement ratio, sand-to-aggregate mass ratio, RBA replacement ratio and mass-weighted water absorption ratio of coarse aggregates. The AI models developed in this study accurately capture the trends of these factors and offer desirable predictive results.

Excavated soil waste is one of the construction wastes generated from building foundation pit excavation, tunnel excavation, channel excavation and other engineering excavations. In order to reuse the excavated soil waste in civil engineering, this paper presents a literature review on the feasibility of using excavated soil waste as fine replacements in cementitious products. With the help of existing experiments reported in the literature, a data-driven analysis was conducted on the mechanical properties of excavated soil-based cement mortar, geopolymer, unfired clay bricks and concrete at normal condition (e.g., room temperature and atmospheric moisture) or after water soaked. The influence of excavated soil replacing river sand (i.e., fine aggregate) or binding materials, curing age, thermal treatment, dry sieving process and stabilization process, etc. on the compressive strength, flexural strength and modulus of elasticity of cementitious materials was discussed. The comparative results show that the excavated soil-based products can achieve acceptable mechanical properties when employing physical approaches (e.g., thermal treatment, sieving and grinding) acting on or mixing stabilizer into excavated soil.

The excavated soil waste and the demolished concrete can be used as the source of natural aggregates’ replacement in concrete. With this purpose, this paper presents two orthogonal experiments (i.e., compression and flexure) to examine the feasibility of jointly using excavated soil and fine recycled concrete aggregate (FRCA) to manufacture unfired clay bricks (UCBs). The water-to-cement ratio (w/c), the cement-to-excavated soil ratio (c/s), the FRCA-to-excavated soil ratio (a/s), and the compound additive content (AC) were selected as the variables in the mix proportion. The test results show that excavated soil & FRCA-based UCBs exhibit the favourable water absorption, bulk density, compressive strength and flexural strength. From the perspective of physical and mechanical properties, the optimal mix proportion is the case of w/c= 0.65, a/s = 0.5, c/s = 0.3, AC= 5%. A grey relational analysis was performed to evaluate the parametric sensitivity of physical properties to four factors. The result revealed that the water absorption is the most sensitive to a/s, and the bulk density is the most sensitive to c/s; in contrast, both of them have the least sensitivity to AC. The prediction models of compressive strength and flexural strength were developed to gain the confidence of recycling excavated soil and FRCA in UCBs. A Scanning Electron Microscope (SEM) was carried out to investigate the mechanism of strength increase of excavated soil & FRCA-based UCBs with different mix proportions, and the result demonstrated that the primary reason of the strength increase is owning to the formation of more hydration products and the tighter internal structure.

The burden of gastrointestinal parasites in zoo animals has serious implications for their welfare and the health of veterinarians and visitors. Zhuyuwan Zoo is located in the eastern suburb of Yangzhou city in eastern China, in which over 40 species of zoo animals are kept. In order to understand the infection status of GI parasites in Zhuyuwan Zoo, a total of 104 fresh fecal samples collected randomly from birds (n = 19), primates (n = 19), and non-primate mammals (n = 66) were analyzed using the saturated saline flotation technique and nylon sifter elutriation and sieving method for eggs/oocysts, respectively. Two Ascaris species were molecularly characterized. The results showed that the overall prevalence of parasitic infection was 42.3% (44/104). The parasitic infection rate in birds, primates, and non-primate mammals were 26.3% (5/19), 31.6% (6/19), and 50.0% (33/66), respectively. A total of 11 species of parasites were identified, namely, Trichostrongylidae, Capillaria sp., Trichuris spp., Strongyloides spp., Amidostomum sp., Toxascaris leonina, Baylisascaris transfuga, Parascaris equorum, Paramphistomum spp., Fasciola spp., and Eimeria spp. Paramphistomum spp. eggs were first detected from the captive Père David’s deer, and Fasciola spp. eggs were first reported from sika deer in zoo in China. A sequence analysis of ITS-2 and cox1 showed that the eggs isolated from the African lion (Panthera leo Linnaeus, 1758) were T. leonina, and the eggs from the brown bear (Ursus arctos Linnaeus, 1758) were B. transfuga. The public health threat posed by these potential zoonotic parasitic agents requires attention. These results lay a theoretical foundation for prevention and control of wild animal parasitic diseases at zoos in China.

Abstract 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 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. Graphical Abstract

Paleoenvironmental information is better preserved in carbonate concretions. In this study, carbonate concretions in the Cretaceous Nenjiang shale, Songliao Basin, were examined to determine whether molecular fossils reflective of the paleoenvironment were better preserved at these sites. Organic and inorganic geochemical characteristics of the concretions and surrounding rocks were analyzed using a series of techniques, including SEM, LA-ICP-MS, GC-MS-MS, and GC-IRMS. The concretions are composed of high content microcrystalline dolomite. The δ13Ccarb and δ18Ocarb values of the concretionary dolomite were significantly higher than those of the surrounding rocks. The dolomite show enrichment in the LREEs and have a negative Eu anomaly. The concretion biomarkers showed distribution characteristics similar to those of surrounding rocks. This suggested that the molecular fossils preserved in concretions were mainly inherited from surrounding rocks. However, the concretions contained more C27 sterane and hopanes, with the hopane/sterane ratio being significantly higher than that of surrounding rocks (1.49 v. 0.86). Moreover, the relative content of 2-methylhopane was 2.4–6.6 times that of the surrounding rocks. This indicated changes in the biological equilibrium of source organisms within and outside the concretions. It was possible that the unstable organic matter at the core increased the bacterial concentration and activity inside the concretions. Both the hydrogen index and biomarker-derived indicators implied that the transformation of organic matter in concretions was minimized when compared with their host rock. The isotope δ13C16-30 was 1‰–3‰ more prevalent in individual N-alkane hydrocarbons in the concretions than in surrounding rocks, likely owing to differences in lithology, bacterial action, and degree of weathering. The study concluded that carbonate concretions could preserve molecular fossils better than the surrounding rocks, and the in-depth organic geochemical analysis of concretions could provide a valuable reference for research into early life forms.

Buildings and Constructions in civil engineering have led to quite a large production of excavated soil wastes. In order to use excavated soil waste, this paper investigated the physical and mechanical properties of excavated soil-based unfired clay bricks (ESUCB) with different mix proportions before and after dry-wet cycles. A total of 108 specimens are separated into nine groups according to the criterial of orthogonal experimental design of four factors (i.e., water-to-cement ratio, cement-to-excavated soil ratio, fine recycled concrete aggregate-to-excavated soil ratio and compound additives content) and three levels. Dry-wet cyclic experiments characterized with 5, 10 and 20 cycles respectively on ESUCB were carried out before conducting the compressive loading test. The test results indicate that increasing the dry-wet cycle number leads to larger mass loss, higher strength reduction rate and reduce the compressive strength of ESUCB. Four strength categories including Grade Ⅰ (higher than 20 MPa), Grade Ⅱ (15–20 MPa), Grade Ⅲ (10–15 MPa) and Grade Ⅳ (5–10 MPa) were divided to provide the specific mix proportion for a certain compressive strength requirement of ESUCB. Variance and range analysis shows that the water-to-cement ratio has the most significant effect on the compressive strength and the smallest one is the additive content. Using the experimental data-driven approach, nonlinear regression equations for compressive strength of ESUCB without and with dry-wet cycles were proposed, respectively.

Abstract Background Eimeria necatrix is one of the most pathogenic parasites, causing high mortality in chickens. Although its genome sequence has been published, the sequences and complete structures of its mRNA transcripts remain unclear, limiting exploration of novel biomarkers, drug targets and genetic functions in E. necatrix. Methods Second-generation merozoites (MZ-2) of E. necatrix were collected using Percoll density gradients, and high-quality RNA was extracted from them. Single-molecule real-time (SMRT) sequencing and Illumina sequencing were combined to generate the transcripts of MZ-2. Combined with the SMRT sequencing data of sporozoites (SZ) collected in our previous study, the transcriptome and transcript structures of E. necatrix were studied. Results SMRT sequencing yielded 21,923 consensus isoforms in MZ-2. A total of 17,151 novel isoforms of known genes and 3918 isoforms of novel genes were successfully identified. We also identified 2752 (SZ) and 3255 (MZ-2) alternative splicing (AS) events, 1705 (SZ) and 1874 (MZ-2) genes with alternative polyadenylation (APA) sites, 4019 (SZ) and 2588 (MZ-2) fusion transcripts, 159 (SZ) and 84 (MZ-2) putative transcription factors (TFs) and 3581 (SZ) and 2039 (MZ-2) long non-coding RNAs (lncRNAs). To validate fusion transcripts, reverse transcription-PCR was performed on 16 candidates, with an accuracy reaching up to 87.5%. Sanger sequencing of the PCR products further confirmed the authenticity of chimeric transcripts. Comparative analysis of transcript structures revealed a total of 3710 consensus isoforms, 815 AS events, 1139 genes with APA sites, 20 putative TFs and 352 lncRNAs in both SZ and MZ-2. Conclusions We obtained many long-read isoforms in E. necatrix SZ and MZ-2, from which a series of lncRNAs, AS events, APA events and fusion transcripts were identified. Information on TFs will improve understanding of transcriptional regulation, and fusion event data will greatly improve draft versions of gene models in E. necatrix. This information offers insights into the mechanisms governing the development of E. necatrix and will aid in the development of novel strategies for coccidiosis control. Graphical Abstract

Histomonas meleagridis is a protozoan parasite that causes histomonosis in gallinaceous birds such as turkeys and chickens. Since the banning and restricted usage of effective drugs such as nitarsone, 80–100% morbidity and mortality occur in turkeys and 20–30% mortality in chickens. New ideas are needed to resolve the re-emergence of histomonosis in poultry. In this study, the α-actinin encoding gene from H. meleagridis was cloned. The 1839-bp gene encoding 612 amnio acids showed close phylogenetic relationships with Trichomonas vaginalis and Tritrichomonas foetus. It was then inserted into the prokaryotic expression vector pET28a(+) and induced with isopropyl-β-D-thiogalactopyranoside. A 73 kDa recombinant protein rHmα-actinin 1 was obtained and purified with a Ni-NTA chromatography column. rHmα-actinin 1 was recognized by mouse anti-rHmα-actinin 1 polyclonal antibody, mouse anti-rHmα-actinin 1 monoclonal antibody, and rehabilitation sera from H. meleagridis infected chickens. Native α-actinin 1 in the total proteins of H. meleagridis can also be detected with mouse anti-rHmα-actinin monoclonal antibody. Immunolocalization assays showed that Hmα-actinin 1 was mainly distributed in the cytoplasm of virulent histomonads JSYZ-D9 and in the peripheral regions (near the plasma membrane) of attenuated histomonads JSYZ-D195. Based on in vivo experiment, when chickens were subcutaneously immunized with rHmα-actinin 1 at 5 and 12 days old and then challenged with H. meleagridis at 19 days old, rHmα-actinin 1 reduced the lesion scores 12 days after infection (31 days old) and increased the body weight gain during the challenged period (19–31 days old). Furthermore, it also strengthened the cellular and humoral immune responses 7 days after the second immunization (19 days old). In conclusion, Hmα-actinin 1 could be used as a candidate antigen to develop vaccines against chicken histomonosis.

Construction and demolition waste (CDW) has been a worldwide issue in the process of urbanization. Chinese government at all levels introduces numerous policies to promote the research and applications of CDW. With respect to financial grants, the National Natural Science Foundation of China (NSFC) has played a huge role in advancing the basic RAC-related researches. This paper presents an information mining-based review on China’s recycling and reuse of CDW from the perspective of NSFC-supported research and research-driven application. A database including 146 approved NSFC projects in the field of RAC was compiled and single factor (i.e., academic title of person in charge, university level/quality, general research topic, university/institute location) influencing the funding ratio of NSFC were discussed. Available technical standards based on the achievements of basic research of RAC were collected, and the real engineering cases from low-rise to high-rise buildings was introduced. It is founded that both the quantity of the approved NSFC projects and the CDW production and utilization in Beijing-Tianjin-Hebei regions, China’s South-East coastal regions and South China regions are commensurate with the level of their regional developments. The guidance for applicants to increase the funding rate of NSFC was suggested after analyzing the future research trend.

Toxoplasma gondii (T. gondii) is an obligate intracellular parasitic protozoan that can cause toxoplasmosis in humans and other endotherms. T. gondii can manipulate the host gene expression profile by interfering with miRNA expression, which is closely associated with the molecular mechanisms of T. gondii-induced brain injury. However, it is unclear how T. gondii manipulates the gene expression of central nervous system (CNS) cells through modulation of miRNA expression in vivo during acute and chronic infection. Therefore, high-throughput sequencing was used to investigate expression profiles of brain miRNAs at 10, 25, and 50 days post-infection (DPI) in pigs infected with the Chinese I genotype T. gondii strain in this study. Compared with the control group 87, 68, and 135 differentially expressed miRNAs (DEMs) were identified in the infected porcine brains at 10, 25, and 50 DPI, respectively. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis showed that a large number significantly enriched GO terms and KEGG pathways were found, and were mostly associated with stimulus or immune response, signal transduction, cell death or apoptosis, metabolic processes, immune system or diseases, and cancers. miRNA–gene network analysis revealed that the crucial connecting nodes, including DEMs and their target genes, might have key roles in the interactions between porcine brain and T. gondii. These results suggest that the regulatory strategies of T. gondii are involved in the modulation of a variety of host cell signaling pathways and cellular processes, containing unfolded protein response (UPR), oxidative stress (OS), autophagy, apoptosis, tumorigenesis, and inflammatory responses, by interfering with the global miRNA expression profile of CNS cells, allowing parasites to persist in the host CNS cells and contribute to pathological damage of porcine brain. To our knowledge, this is the first report on miRNA expression profile in porcine brains during acute and chronic T. gondii infection in vivo. Our results provide new insights into the mechanisms underlying T. gondii-induced brain injury during different infection stages and novel targets for developing therapeutic agents against T. gondii.

In the field of construction, the surface of architectural aluminum formwork for concrete pouring will remain the concrete adhesion layer of heterogeneous composite structures. In view of the difficulty of removing the concrete adhesion layer, we studied the technology and mechanism of removing the concrete adhesion layer by laser cleaning technology in this paper. We analyzed the composition and distribution characteristics of residual concrete on the surface of architectural aluminum formwork, set up a laser cleaning test system, carried out laser cleaning experiments on the concrete layer on the surface of architectural aluminum formwork under different storage times, and analyzed the mechanism of removing the concrete adhesion layer by laser cleaning. The experimental results showed that the residual time of concrete will affect the quality and efficiency of laser cleaning concrete residue on the surface of architectural aluminum formwork for concrete pouring. For concrete residues with short residual time, lasering can achieve efficient and high-quality cleaning. A nanosecond pulsed laser could strengthen the surface hardness of the aluminum alloy template during cleaning, which is helpful in improving the durability of the aluminum alloy template. The main mechanisms of laser cleaning to remove the concrete adhesion layer on the surface of architectural aluminum formwork is that the bubbles and water bubbles in the loose structure of concrete instantly absorb high-energy laser and make the concrete aggregate continuously air-burst. This paper provides technological and theoretical support for the application of laser cleaning technology to remove residual concrete on the surface of architectural aluminum formwork for concrete pouring in the field of construction.

Abstract Background Toxoplasma gondii is an obligate intracellular parasite that infects humans and other warm-blooded animals. Previous quantitative proteomic analyses of infected host cells revealed that the expression of many host proteins is modulated by T. gondii infection. However, at present limited data are available on the differentially expressed miRNAs (DEMs) associated with the pathology and host immune responses induced by acute and chronic infection with T. gondii in pigs in vivo. In this study, high-throughput sequencing was used to investigate expression profiles of spleen miRNAs at 10, 25 and 50 days post-infection (DPI) in pigs infected with Chinese I genotype strain T. gondii isolated from a dead pig. Results When compared to the control group, 34, 6 and 86 DEMs were found in spleens of infected pigs at 10, 25 and 50 DPI, respectively. Gene Ontology (GO) enrichment analysis of the target genes of DEMs showed that no GO terms were enriched at 25 DPI, whereas 28 and 241 GO terms, of which two and 215 were sample-specific, were significantly enriched at 10 and 50 DPI, respectively. The top 20 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways of the target genes of DEMs included signal transduction, immune system, metabolism and diseases. miRNA–gene network analysis revealed that the DEMs played important roles in the host immune response to T. gondii infection by modulating expression levels of cellular immunity-related cytokines and immune-related C-type lectins. Conclusion Our results not only showed that host miRNA expression is altered by T. gondii but also revealed differences in the regulation of key biological processes and pathways involved in host responses to acute versus chronic T. gondii infection. This will aid future research into miRNA-target interactions during T. gondii infection in pigs and the development of novel therapies against T. gondii.

Abstract Background Eimeria is a common genus of apicomplexan parasites that infect diverse vertebrates, most notably poultry, causing serious disease and economic losses. Eimeria species have complex life-cycles consisting of three developmental stages. However, the molecular basis of the Eimeria reproductive mode switch remains an enigma. Methods Total RNA extracted from second- (MZ-2) and third-generation merozoites (MZ-3) of Eimeria necatrix was subjected to transcriptome analysis using RNA sequencing (RNA-seq) followed by qRT-PCR validation. Results A total of 6977 and 6901 unigenes were obtained from MZ-2 and MZ-3, respectively. Approximately 2053 genes were differentially expressed genes (DEGs) between MZ-2 and MZ-3. Compared with MZ-2, 837 genes were upregulated and 1216 genes were downregulated in MZ-3. Approximately 95 genes in MZ-2 and 48 genes in MZ-3 were further identified to have stage-specific expression. Gene ontology category and KEGG analysis suggested that 216 upregulated genes in MZ-2 were annotated by 70 GO assignments, 242 upregulated genes were associated with 188 signal pathways, while 321 upregulated genes in MZ-3 were annotated by 56 GO assignments, 322 upregulated genes were associated with 168 signal pathways. The molecular functions of upregulated genes in MZ-2 were mainly enriched for protein degradation and amino acid metabolism. The molecular functions of upregulated genes in MZ-3 were mainly enriched for transcriptional activity, cell proliferation and cell differentiation. Conclusions To the best of our knowledge, this is the first RNA-seq data study of the MZ-2 and MZ-3 stages of E. necatrix; it demonstrates a high number of differentially expressed genes between the MZ-2 and MZ-3 of E. necatrix. This study forms a basis for deciphering the molecular mechanisms underlying the shift from the second to third generation schizogony in Eimeria. It also provides valuable resources for future studies on Eimeria, and provides insight into the understanding of reproductive mode plasticity in different Eimeria species.

In order to study the quality control and evaluation methods of self-compacting concrete (SCC) pumping process in Shenzhen-Zhongshan Bridge and similar projects, sample test is performed on self-compacting concrete mixture collected from the pumping field of the E1-E4 steel shell immersed tube; Then a database base on relationship between the variation parameters and the target performance is established. On this basis, the Grey system theory is adopted to analyze the parameter sensitivity of the SCC pumping performance to the different kinds of variables. The results show that variables are related to target performance and some of the variables have a significant influence. Using the powerful data mining capability of support-vector machine and Bayesian statistical inference in the case of uncertain exact mathematical relationship between independent variables and dependent variables, implicit and explicit prediction models of variation of SCC pumping performance are respectively established by pumping distance, number of elbows, pumping time and environmental temperature as the control parameters. Finally, the comparisons between the measured data and calculation result prove that both models have good prediction accuracy and stability.

Abstract Background Gametocyte proteins of Eimeria (E.) spp. are important components of the oocyst wall and some have been used to develop transmission-blocking vaccines against avian coccidiosis. Methods Total RNA isolated from E. necatrix gametocytes was utilized as templates for RT-PCR amplification and sequencing of cDNA encoding a gametocyte protein using gene-specific primers. The cDNA was cloned into the bacterial expression vector pET28a(+) and expressed in E. coli BL21 cells. The antigenicity of the recombinant gametocyte protein and its localization in different E. necatrix life-cycle stages were determined by western blot and indirect immunofluorescence analyses, respectively. Results A 731-nucleotide sequence of cDNA [GenBank: KF649255] of E. necatrix had 97.7% identity to that of Etgam22 of E. tenella. The cDNA ORF encoded a 186-amino acid protein containing a histidine-proline-rich region. The recombinant gametocyte protein (rEnGAM22) was predominately expressed in the insoluble inclusion body and recognized by antiserum from chickens immunized with oocysts of E. necatrix, E. maxima and E. tenella. A specific antibody to the rEnGAM22 protein recognized the wall-forming bodies in macrogametocytes and the walls of oocysts and sporocysts. Conclusions The gene cloned from E. necatrix gametocytes is an ortholog to Etgam22 of E. tenella and presents a potential target for future recombinant subunit vaccines against coccidiosis.

African trypanosomes are extracellular protozoan parasites causing a chronic debilitating disease associated with a persistent inflammatory response. Maintaining the balance of the inflammatory response via downregulation of activation of M1-type myeloid cells was previously shown to be crucial to allow prolonged survival. Here we demonstrate that infection with African trypanosomes of IL-27 receptor-deficient (IL-27R-/-) mice results in severe liver immunopathology and dramatically reduced survival as compared to wild-type mice. This coincides with the development of an exacerbated Th1-mediated immune response with overactivation of CD4+ T cells and strongly enhanced production of inflammatory cytokines including IFN-γ. What is important is that IL-10 production was not impaired in infected IL-27R-/- mice. Depletion of CD4+ T cells in infected IL-27R-/- mice resulted in a dramatically reduced production of IFN-γ, preventing the early mortality of infected IL-27R-/- mice. This was accompanied by a significantly reduced inflammatory response and a major amelioration of liver pathology. These results could be mimicked by treating IL-27R-/- mice with a neutralizing anti-IFN-γ antibody. Thus, our data identify IL-27 signaling as a novel pathway to prevent early mortality via inhibiting hyperactivation of CD4+ Th1 cells and their excessive secretion of IFN-γ during infection with African trypanosomes. These data are the first to demonstrate the essential role of IL-27 signaling in regulating immune responses to extracellular protozoan infections.