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Two prototypical genotoxicants, benzo[a]pyrene (B[a]P) and colchicine (COL), were selected as model compounds to deduce their quantitative genotoxic dose–response relationship at low doses in a multi-endpoint genotoxicity assessment platform. Male Sprague-Dawley rats were treated with B[a]P (2.5–80 mg/kg bw/day) and COL (0.125–2 mg/kg bw/day) daily for 28 days. The parameters included were as follows: comet assay in the peripheral blood and liver, Pig-a gene mutation assay in the peripheral blood, and micronucleus test in the peripheral blood and bone marrow. A significant increase was observed in Pig-a mutant frequency in peripheral blood for B[a]P (started at 40 mg/kg bw/day on Day 14, started at 20 mg/kg bw/day on Day 28), whereas no statistical difference for COL was observed. Micronucleus frequency in reticulocytes of the peripheral blood and bone marrow increased significantly for B[a]P (80 mg/kg bw/day on Day 4, started at 20 mg/kg bw/day on Days 14 and 28 in the blood; started at 20 mg/kg bw/day on Day 28 in the bone marrow) and COL (started at 2 mg/kg bw/day on Day 14, 1 mg/kg bw/day on Day 28 in the blood; started at 1 mg/kg bw/day on Day 28 in the bone marrow). No statistical variation was found in indexes of comet assay at all time points for B[a]P and COL in the peripheral blood and liver. The dose–response relationships of Pig-a and micronucleus test data were analyzed for possible point of departures using three quantitative approaches, i.e., the benchmark dose, breakpoint dose, and no observed genotoxic effect level. The practical thresholds of the genotoxicity of B[a]P and COL estimated in this study were 0.122 and 0.0431 mg/kg bw/day, respectively, and our results also provided distinct genotoxic mode of action of the two chemicals.

A sequential batch reactor (SBR) was used to enrich and produce a self-protected CaCO3-precipitating biomass powder (CPB) that hydrolzes urea and reduces nitrate. The self-protected CPB, consisting of 50% biomass and 50% inorganic matter, was incorporated into a mortar and its self-healing performance was assessed. Crack closure observations showed that specimens containing CPB showed higher crack healing ratio in complete water immersion, while Bacillus sphaericus spores containing samples only exhibited crack healing under cyclic wet-dry incubation. During corrosion monitoring, CPB addition revealed a corrosion inhibition effect equivalent to that achieved by the addition of the chemical corrosion inhibitor nitrite (0.7 w/w% cement weight). Overall, CPB is a suitable self-healing bio-additive for effective crack closure in cementitious composites used in immersed structural elements. It is compatible with the cementitious matrix and resulted in promising results concerning crack healing and corrosion inhibition.

Presupposition, as a pragmatic concept, plays an important role in discourse construction. Pragmatic presupposition works inside and outside ironic discourse to help maintain cohesion and coherence as well as direction and integrity of it. This paper presents pragmatic analyses on irony selected from Pride and Prejudice with different functions of presuppositions from social standards, collusive agreement, physical situation, and previous circumstances, and hopes to provide a different way of theorizing irony and ironic discourse.

Background. The fruits ofMalania oleifera Chun & S. K. Leehave been highly sought after medically because its seeds have high oil content (>60%), especially the highest known proportion of nervonic acid (>55%).Objective of the Study.The objective was to explore the effects of different doses ofMalania oleifera Chunoil(MOC oil) on the learning and memory of mice and to evaluate whether additional DHA algae oil and vitamin E could help MOC oil improve learning and memory and its possible mechanisms.Methods. After 30 days of oral administration of the relevant agents to mice, behavioral tests were conducted as well as detection of oxidative stress parameters (superoxide dismutase, malondialdehyde, and glutathione peroxidase) and biochemical indicators (acetylcholine, acetyl cholinesterase, and choline acetyltransferase) in the hippocampus.Results. Experimental results demonstrated that MOC oil treatment could markedly improve learning and memory of mouse models in behavioral experiments and increase the activity of GSH-PX in hippocampus and reduce the content of MDA, especially the dose of 46.27 mg/kg. The addition of DHA and VE could better assist MOC oil to improve the learning and memory, and its mechanism may be related to the inhibition of oxidative stress and restrain the activity of AChE and also increase the content of ACh.Conclusion. Our results demonstrated that MOC oil treatment could improve learning and memory impairments. Therefore, we suggest that MOC oil is a potentially important resource for the development of nervonic acid products.

The main goal of the study was to investigate the genotoxic response of N-ethyl-N-nitrosourea (ENU) and ethyl methanesulfonate (EMS) at low doses in a multi-endpoint genotoxicity assessment platform in rats and to derive potential thresholds and related metrics. Male Sprague–Dawley rats were treated by daily oral gavage for 28 consecutive days with ENU (0.25 ~ 8 mg/kg bw) and EMS (5 ~ 160 mg/kg bw), both with six closely spaced dose levels. Pig-a gene mutation assay, micronucleus test, and comet assay were performed in several timepoints. Then, the dose–response relationships were analyzed for possible points of departure (PoD) using the no observed genotoxic effect level and benchmark dose (BMD) protocols with different critical effect sizes (CES, 0.05, 0.1, 0.5, and 1SD). Overall, dose-dependent increases in all investigated endpoints were found for ENU and EMS. PoDs varied across genetic endpoints, timepoints, and statistical methods, and selecting an appropriate lower 95% confidence limit of BMD needs a comprehensive consideration of the mode of action of chemicals, the characteristics of tests, and the model fitting methods. Under the experimental conditions, the PoDs of ENU and EMS were 0.0036 mg/kg bw and 1.7 mg/kg bw, respectively.

Swine coronavirus–porcine epidemic diarrhea virus (PEDV) with specific susceptibility to pigs has existed for decades, and recurrent epidemics caused by mutant strains have swept the world again since 2010. In this study, single-cell RNA sequencing was used to perform for the first time, to our knowledge, a systematic analysis of pig jejunum infected with PEDV. Pig intestinal cell types were identified by representative markers and identified a new tuft cell marker, DNAH11. Excepting enterocyte cells, the goblet and tuft cells confirmed susceptibility to PEDV. Enrichment analyses showed that PEDV infection resulted in upregulation of cell apoptosis, junctions, and the MAPK signaling pathway and downregulation of oxidative phosphorylation in intestinal epithelial cell types. The T cell differentiation and IgA production were decreased in T and B cells, respectively. Cytokine gene analyses revealed that PEDV infection downregulated CXCL8, CXCL16, and IL34 in tuft cells and upregulated IL22 in Th17 cells. Further studies found that infection of goblet cells with PEDV decreased the expression of MUC2, as well as other mucin components. Moreover, the antimicrobial peptide REG3G was obviously upregulated through the IL33-STAT3 signaling pathway in enterocyte cells in the PEDV-infected group, and REG3G inhibited the PEDV replication. Finally, enterocyte cells expressed almost all coronavirus entry factors, and PEDV infection caused significant upregulation of the coronavirus receptor ACE2 in enterocyte cells. In summary, this study systematically investigated the responses of different cell types in the jejunum of piglets after PEDV infection, which deepened the understanding of viral pathogenesis.

There are intense interests in discovering proregenerative medicine leads that can promote cardiac differentiation and regeneration, as well as repair damaged heart tissues. We have combined zebrafish embryo-based screens with cardiomyogenesis assays to discover selective small molecules that modulate heart development and regeneration with minimal adverse effects. Two related compounds with novel structures, named as Cardiomogen 1 and 2 (CDMG1 and CDMG2), were identified for their capacity to promote myocardial hyperplasia through expansion of the cardiac progenitor cell population. We find that Cardiomogen acts as a Wnt inhibitor by targeting β-catenin and reducing Tcf/Lef-mediated transcription in cultured cells. CDMG treatment of amputated zebrafish hearts reduces nuclear β-catenin in injured heart tissue, increases cardiomyocyte (CM) proliferation, and expedites wound healing, thus accelerating cardiac muscle regeneration. Importantly, Cardiomogen can alleviate the functional deterioration of mammalian hearts after myocardial infarction. Injured hearts exposed to CDMG1 display increased newly formed CMs and reduced fibrotic scar tissue, which are in part attributable to the β-catenin reduction. Our findings indicate Cardiomogen as a Wnt inhibitor in enhancing injury-induced CM proliferation and heart regeneration, highlighting the values of embryo-based small molecule screens in discovery of effective and safe medicine leads.

miR-181a is downregulated in leukemia and affects its progression, drug resistance, and prognosis. However, the exact mechanism of its targets in leukemia, particularly in chronic myelogenous leukemia (CML), has not previously been established. Here, we use a multi-omics approach to demonstrate that protein tyrosine phosphatase, receptor type, f polypeptide, leukocyte common antigen (LAR) interacting protein (liprin), alpha 1 (PPFIA1) is a direct target for miR-181a in CML. Phospho-array assay shows that multiple phosphorylated proteins, particularly KIT signaling molecules, were downregulated in PPFIA1 inhibition. Additionally, PPFIA1 bound PARP1, a common molecule downstream of both PPFIA1 and BCR/ABL, to upregulate KIT protein through activation of nuclear factor kappa B (NF-κB)-P65 expression. Targeted inhibition of PPFIA1 and PARP1 downregulated c-KIT level, inhibited CML cell growth, and prolonged mouse survival. Overall, we report a critical regulatory miR-181a/PPFIA1/PARP1/NF-κB-P65/KIT axis in CML, and our preclinical study supports that targeted PPFIA1 and PARP1 may serve as a potential CML therapy. Keywords: CML, multi-omics, miR-181a, PPFIA1, PARP1, siRNA treatment

Deoxynivalenol (DON), zearalenone (ZEN), and fumonisin B1 (FB1), as the main mycotoxins contaminating rice, often coexist in food. Thus, we have measured the genotoxicity of the three rice fungal contaminants, singly and in different combinations, with a 28-day multi-endpoint (Pig-a assay + in vivo micronucleus [MN] test + comet assay) genotoxicity platform. Male Sprague-Dawley rats received the agents orally via gavage for 28 consecutive days, before performing the abovementioned tests. Results indicated that low dose of a single mycotoxin did not show significant genotoxicity. However, some of these mycotoxins in combination induced significant genotoxicity in the peripheral blood and tissues, at sacrifice. In the peripheral blood, the binary combination of DON and FB1 significantly induced MN. In the liver, ZEN might aggravate the DNA-damaging effects of DON and FB1. Therefore, the genotoxicity of sub-chronic exposure to mycotoxins in combination cannot be ignored.

Background: African Swine Fever Virus (ASFV) has been identified as the agent of ASF, which has resulting in a mortality rate of nearly 100% in domestic pigs worldwide. Protein p22 encoded by pKP177L was reported to be localized at the inner envelope of the virus, while the function of p22 remains unclear.Methods: Protein p22 interacted proteins of the host were immune-precipitated and identified by Liquid Chromatography Mass Spectrometry, analyzed by Go terms and KEGG pathways. Results: Numerous cellular proteins in 293-T that interacted with p22 protein were identified. These interacted proteins were related to the biological processes of binding, cell structure, signal transduction, cell adhesion, etc., and their interactions. At the same time, the interacted proteins participated in several KEGG pathways like ribosome, splicesome, etc. The key proteins in PPI network were closely related to actin filament organization and movement, resulting in affecting the process of phagocytosis and endocytosis. Conclusions: The identified high number of proteins interacted with p22 and as a large database should be very useful to elucidate the function of p22 in the near future, and lay the foundation for elucidating the mechanism of ASFV.

Teleost zebrafish and neonatal mammalian hearts exhibit the remarkable capacity to regenerate through dedifferentiation and proliferation of pre-existing cardiomyocytes (CMs). Although many mitogenic signals that stimulate zebrafish heart regeneration have been identified, transcriptional programs that restrain injury-induced CM renewal are incompletely understood. Here, we report that mutations in gridlock (grl; also known as hey2), encoding a Hairy-related basic helix-loop-helix transcriptional repressor, enhance CM proliferation and reduce fibrosis following damage. In contrast, myocardial grl induction blunts CM dedifferentiation and regenerative responses to heart injury. RNA sequencing analyses uncover Smyd2 lysine methyltransferase (KMT) as a key transcriptional target repressed by Grl. Reduction in Grl protein levels triggered by injury induces smyd2 expression at the wound myocardium, enhancing CM proliferation. We show that Smyd2 functions as a methyltransferase and modulates the Stat3 methylation and phosphorylation activity. Inhibition of the KMT activity of Smyd2 reduces phosphorylated Stat3 at cardiac wounds, suppressing the elevated CM proliferation in injured grl mutant hearts. Our findings establish an injury-specific transcriptional repression program in governing CM renewal during heart regeneration, providing a potential strategy whereby silencing Grl repression at local regions might empower regeneration capacity to the injured mammalian heart., Highlighted Article: Novel mechanisms of the Grl-Smyd2 network govern vertebrate CM renewal and heart regeneration, which might be relevant in developing strategies for regeneration interventions in humans.

We have measured the toxicity and genotoxicity of 2-methylfuran, which is formed in foods during thermal processing. The agent was administered by oral gavage to male Sprague-Dawley rats, daily for 28 days, before performing general toxicology analysis and the following genotoxicity tests: comet assay (peripheral blood, liver); Pig-a gene mutation assay (peripheral blood); micronucleus test (peripheral blood, bone marrow). Liver was the primary target organ; histological changes (oval cell hyperplasia) were observed but without significant changes in serum enzyme markers. For hepatotoxicity, the no-observed-adverse-effect level was 5 mg/kg bw/d. Histopathological changes were also seen in the bone marrow. Genotoxicity assays were uniformly negative.

The fruits ofAfter 30 days of oral administration of the relevant agents to mice, behavioral tests were conducted as well as detection of oxidative stress parameters (superoxide dismutase, malondialdehyde, and glutathione peroxidase) and biochemical indicators (acetylcholine, acetyl cholinesterase, and choline acetyltransferase) in the hippocampus.Experimental results demonstrated that MOC oil treatment could markedly improve learning and memory of mouse models in behavioral experiments and increase the activity of GSH-PX in hippocampus and reduce the content of MDA, especially the dose of 46.27 mg/kg. The addition of DHA and VE could better assist MOC oil to improve the learning and memory, and its mechanism may be related to the inhibition of oxidative stress and restrain the activity of AChE and also increase the content of ACh.Our results demonstrated that MOC oil treatment could improve learning and memory impairments. Therefore, we suggest that MOC oil is a potentially important resource for the development of nervonic acid products.

Porcine circovirus type 2 (PCV2) is an important pathogen of the current pig industry. The Wnt signalling pathway plays an important role in the growth of young animals. In this study, we mainly elucidated the relationship between PCV2 and the Wnt signalling pathway. In an in vivo experiment in mice, we demonstrated the downregulatory effects of PCV2 infection on expression levels of downstream components of the Wnt signalling pathway. Weight loss in mice was reversed by activating the Wnt signalling pathway, and the body weight was still significantly higher than that in mice infected with PCV2. We detected levels of growth hormone (GH) in the liver and sera, which showed that GH was also downregulated in mice challenged with PCV2. Lithium chloride, the activator of Wnt signalling, upregulated GH, albeit to a significantly lesser degree than that in corresponding non-stimulated mock mice. In vitro studies showed that PCV2 infection downregulated protein expression of β-catenin and mRNA expression of matrix metallopeptidase-2 (Mmp2), downregulated protein expression of β-catenin in the cytoplasm and nucleus, and reduced the activity of the TCF/LEF promoter, demonstrating that PCV2 inhibited activation of the Wnt signalling pathway in vitro. Finally, we found that Rep protein of PCV2 might be responsible for the inhibitory effect.

The micronucleus test (MNT) is the most widely applied short-term assay to detect clastogens or spindle disruptors. The use of flow cytometry (FCM) has been reported for micronucleated erythrocytes scoring in peripheral blood. The aim of this study was to develop a novel and practical protocol for MNT in rat peripheral blood by FCM, with the method validation. CD71-fluorescein isothiocyanate and DRAQ5 were adopted for the fluorescent staining of proteins and DNA, respectively, to detect micronuclei. To validate the method, groups of male Sprague-Dawley rats (five per group) received two oral gavage doses at 0 and 24 h of six chemicals (four positive mutagens: ethyl methanesulphonate [EMS], cyclophosphamide [CP], colchicine [COL], and ethyl nitrosourea [ENU]; two nongenotoxic chemicals: sodium saccharin and eugenol). Blood samples were collected from the tail vein before and on the five continuous days after treatments; all of which were analyzed for micronuclei presence by both the manual (Giemsa staining) and FCM methods. The FCM-based method consistently demonstrated highly sensitive responses for micronucleus detection at all concentrations and all time points for EMS, CP, COL, and ENU. Sodium saccharin and eugenol could be identified as negative in this protocol. Results obtained with the FCM-based method correlated well with the micronucleus frequencies (r = 0.659-0.952), and the proportion of immature erythrocytes (r = 0.915-0.981) tested by Giemsa staining. The method reported here, with easy operation, low background, and requirement for a regular FCM, could be an efficient system for micronucleus scoring.

The process of microbially induced calcite precipitation (MICP) is widely used recently in construction engineering in improving compressive strength, durability and self-healing of building materials and culture heritages. However, most of researches to date have concentrated on prokaryotic systems despite of associated limitation of urease-positive bacteria in biocementation. In the present study, we exploited the role of one urease-positive fungal strain Penicillium chrysogenum CS1 for the first time in biocementation of sand in column to produce sandstone of significant compressive strength. Further, the research provided understanding of the involved mechanisms and advantages of fungal-mediated production of calcite in cementing sand granules over same process using bacteria.

This study was designed to determine the genotoxicity of 2-methylfuran based on a multi-endpoint genotoxicity test system.The SPF-grade male SD rats(n = 30) were randomized to six treatment groups, i. e. 4 treatment groups(25, 50, 100 and 150 mg/kg), a control group(vegetable oil) and a positive groups(N-ethyl-N-nitrosourea, 80 mg/kg). All treatments were administrated by gavage for continuous 3 days. Tail vein blood for comet assay was collected at 3 h after the final administration. Pig-a gene mutation assays were performed on days 0(one day before gavage), 14 and 28. Micronucleus tests in peripheral blood using flow cytometry were performed on days 0 and 4.A statistically significant increase in tail intensity was observed at 150 mg/kg for peripheral blood in comet assay. There was no significant difference among the groups in mutant cell frequency of erythrocytes and reticulocytes at 2 timepoints in Pig-a gene mutation assay, and no significant difference among the groups in the frequency of micronucleus in micronucleus test.The result of genotoxicity tests suggested that 2-methylfuran was probably not mutagenic in vivo after acute exposure.

Background The hypertension prevalence rate is increasing but the control rate is unsatisfactory. Nurse-led healthcare may be an effective way to improve outcomes for hypertensive patients but more evidence is required especially at the community level. Objective This study aims to establish a nurse-led hypertension management model and to test its effectiveness at the community level. Design A single-blind, randomized controlled trial was performed in an urban community healthcare center in China. Hypertensive patients with uncontrolled blood pressure (systolic blood pressure ≥140mmHg and/or diastolic blood pressure ≥90mmHg) were randomly allocated into two groups: the study group (n=67) and the control group (n=67). The nurse-led hypertension management model included four components (delivery system design, decision support, clinical information system and self-management support). Patients in the control group received usual care. Patients in the study group received a 12-week period of hypertension management. The patient outcomes, which involved blood pressure, self-care behaviors, self-efficacy, quality of life and satisfaction, were assessed at three time points: the baseline, immediately after the intervention and 4 weeks after the intervention. Results After the intervention, the blood pressure of patients in the study group decreased significantly compared to those in the control group, and the mean reduction of systolic/diastolic blood pressure in the study and control groups was 14.37/7.43mmHg and 5.10/2.69mmHg, respectively ( p 0.01 ). In addition, patients in the study group had significantly greater improvement in self-care behaviors than those in the control group (p 0.01) . The study group had a higher level of satisfaction with hypertensive care than the control group (p . No statistically significant difference in self-efficacy and quality of life was detected between the two groups after the intervention. Conclusions The nurse-led hypertension management model is feasible and effective in improving the outcomes of patients with uncontrolled blood pressure at the community level.

Techniques for bacteria based self-healing concrete are currently limited to indirect bacterial activity measurements and visual crack closing effects after self-healing has occurred. In the present study, direct and reliable bacteria survival measurements after encapsulation in protective carriers were carried out by utilizing microcalorimetry analysis. After further incorporation in mortar oxygen consumption was determined by use of an optical oxygen sensing system. The microscope analysis showed that a 90% sporulation of Bacillus sphaericus was achieved after 24 h incubation. Microcalorimeter measurements detected the heat generation from microbial activity on the third day of incubation which provides direct evidence that bacteria can be activated after encapsulation inside the hydrogel. The application of the O2 sensing film on cracked mortar showed a high oxygen concentration reduction (66%) near the crack opening. This confirmed the activation of B. sphaericus and use of oxygen as an electron acceptor for their metabolism after incorporation inside mortar specimens.

The role of industrial byproduct as supplementary cementitious material to partially replace cement has greatly contributed to sustainable environment. Metakaolin (MK), one of such byproduct, is widely used to partial replacement of cement; however, during cement replacement at high percentage, it may not be a good choice to improve the strength of concrete. Thus, in the present study, biocement, a product of microbially induced carbonate precipitation is utilized in MK-modified cement mortars to improve its compressive strength. Despite of cement replacement with MK as high as 50%, the presented technology improved compressive strength of mortars by 27%, which was still comparable to those mortars with 100% cement. The results proved that biomineralization could be effectively used in reducing cement content without compromising compressive strength of mortars. Biocementation also reduced the porosity of mortars at all ages. The process was characterized by SEM-EDS to observe bacterially-induced carbonate crystals and FTIR spectroscopy to predict responsible bonding in the formation of calcium carbonate. Further, XRD analysis identified bio/minerals formed in the MK-modified mortars. The study also encourages combining biological role in construction engineering to solve hazardous nature of cement and at same time solve the disposal problem of industrial waste for sustainable environment.

Objective Routine fasting (12 h) is always applied before laparoscopic cholecystectomy, but prolonged preoperative fasting causes thirst, hunger, and irritability as well as dehydration, low blood glucose, insulin resistance and other adverse reactions. We assessed the safety and efficacy of a shortened preoperative fasting period in patients undergoing laparoscopic cholecystectomy. Methods We searched PubMed, Embase and Cochrane Central Register of Controlled Trials up to 20 November 2015 and selected controlled trials with a shortened fasting time before laparoscopic cholecystectomy. We assessed the results by performing a meta-analysis using a variety of outcome measures and investigated the heterogeneity by subgroup analysis. Results Eleven trials were included. Forest plots showed that a shortened fasting time reduced the operative risk and patient discomfort. A shortened fasting time also reduced postoperative nausea and vomiting as well as operative vomiting. With respect to glucose metabolism, a shortened fasting time significantly reduced abnormalities in the ratio of insulin sensitivity. The C-reactive protein concentration was also reduced by a shortened fasting time. Conclusions A shortened preoperative fasting time increases patients’ postoperative comfort, improves insulin resistance, and reduces stress responses. This evidence supports the clinical application of a shortened fasting time before laparoscopic cholecystectomy.

Porcine circovirus type2 (PCV2) is a member of the circoviridae family. PCV2 was identified as the main pathogen of postweaning multisystemic wasting syndrome (PMWS) in weaned piglets and causes massive economic loss. Basigin, is a transmembrane glycoprotein belonging to the immunoglobulin superfamily; which is also a receptor for cyclophilins. CyP belongs to the immunophilin family that has peptidyl-prolyl cis-trans isomerase activity. Basigin-CyP interaction affects the replication stages of several viruses. In this study, we found that Basigin could elevate the replication of PCV2, and the Basigin only affected the replication stage rather than adsorption or endocytosis stages. In addition, the ligands of Basigin, CyPA and CyPB also elevated the replication of PCV2. Basigin-CyP interation was necessary for elevating PCV2 replication; At last, CyPs were proved to promote the replication of PCV2 by activating ERK signaling.

Background CXCR3, a G-protein coupled chemokine receptor, has been shown to play a critical role in recruiting inflammatory cells into lungs in several studies. However, its roles in polymicrobial septic acute lung injury (ALI) is yet unknown. Therefore, the purpose of this study was to elucidate the protective effects of CXCR3 blockade on pulmonary microvascular endothelial cells (PMVECs) in septic ALI and explore potential mechanisms. Materials and methods ALI was induced by polymicrobial sepsis through cecal ligation and puncture surgery. The expression of CXCR3 on pulmonary microvascular endothelial cells was measured 24 h after cecal ligation and puncture surgery. In addition, the protective effects of neutralizing antibody were detected, including protein concentration, inflammation cell counts, lung ​wet-to-dry ratio, and lung damages. In human umbilical vein endothelial cells (HUVECs) culture condition, CXCR3 expression was measured after exposure to tumor necrosis factor-α. The permeability and apoptosis ratio were detected through CXCR3 gene silencing on HUVECs. The p38 mitogen-activated protein kinase (MAPK) was analyzed with Western blot. Results CXCR3 expression was upregulated both in vivo and in vitro. After CXCR3 neutralizing antibody administrated intraperitoneally, the protein concentration, inflammatory cell counts in BALF and lung ​wet-to-dry ratio were decreased significantly, as well as the lung tissue damages. In vitro, CXCR3 gene silencing inhibited tumor necrosis factor-α and CXCL10-induced hyperpermeability and apoptosis in HUVECs. In addition, p38 mitogen-activated protein kinase activation was essential for CXCR3-mediated apoptosis. Conclusions CXCR3 blockade exerts protective effects on ALI at least partly by inhibiting endothelial cells apoptosis and decreasing the leakage of protein-rich fluid and inflammatory cells. Blockade of CXCR3 may be a promising therapeutic strategy for severe sepsis-induced ALI.

The main objective of this study is to demonstrate the potential of microbial induced calcite precipitation (MICP) as rapid and effective process in the synthesis of fluorescent cadmium sulfide nanoparticles. A urease-producing bacterial isolate AX5, identified as Citrobacter braakii, from calcite rich shells of abalone that could tolerate cadmium at the concentration of 250 mg/l was used. In order to investigate the role of MICP in enhancing the synthesis of CdS nanoparticles, the biomimetic synthesis was carried out by C. braakii in modified nutrient broth without amending any additional organic or toxic surfactants back into the environment. The large amount of monodispersed CdS nanoparticles was harvested and found to bind with biocalcite. XRD analysis confirmed the presence of CdS nanoparticles bound with biocalcite minerals in the precipitation. HRTEM and SEM micrographs showed spherical CdS nanoparticles of particle size in the range of 50–100 nm. The other characteristics of harvested CdS nanoparticles were studied with UV–vis, PL spectra, FTIR, and fluorescence microscopy. The findings of present study have important insights into the application of MICP as a promising mechanism for the biosynthesis of CdS nanoparticles. Keywords: Biocalcite, Nanoparticles, Citrobacter braakii, Urease, CdS

Porcine circovirus type 2 (PCV2) causes significant economic losses to the swine industry worldwide. Heat shock proteins (Hsps) can be used as modulators to enhance both innate and adaptive immune responses. In the present study, recombinant baculoviruses expressing the PCV2Cap protein and the N-terminal 22-370 amino acids of porcine Gp96 (Gp96N), Hsp90, and Hsp70 (rBac-cap/Gp96N, rBac-cap/Hsp90 and rBac-cap/Hsp70, respectively) were constructed and the immune responses were examined in mice and piglets. The mouse experiments showed that rBac-cap/Gp96N increased the titers of specific anti-PCV2 neutralizing antibodies, proliferative responses of peripheral blood mononuclear cells (PBMCs) and IFN-γ levels compared to rBac-cap/Hsp90, rBac-cap/Hsp70, or rBac-cap. The pig experiments showed that the levels of anti-PCV2 antibody, proliferative responses of PBMCs, and IFN-γ in the rBac-cap/Gp96N groups were increased compared to those in rBac-cap group. There were no clear clinical signs of infection following PCV2 challenge in pigs inoculated with recombinant rBac-cap/Gp96N and rBac-cap, and the relative daily weight gains were higher than those in the challenge control (CC) group. The pathological lesions, extent of viremia, and viral loads of the vaccinated groups were milder than those in the CC group. Meanwhile, the extent of viremia and viral load present in the rBac-cap/Gp96N group were significantly lower than those in the rBac-cap group. These results indicated that porcine Gp96N effectively increased the humoral and cell-mediated immune responses of PCV2Cap. Gp96N presents an attractive adjuvant or immunotargeting strategy to enhance the protective efficacy of PCV2 subunit vaccines in swine.

Alpha-linolenic acid (ALA), an important component of polyunsaturated fatty acids (PUFAs), possesses potent anti-inflammatory properties. To date, the effects of ALA on acute lung injury (ALI) remains unknown. This study was designed to investigate the potential protective effects of ALA on LPS-induced ALI and the underpinning mechanisms. An animal model of ALI was established via intratracheally injection of lipopolysaccharide (LPS, 1 mg/kg). We found that lung wet/dry weight ratio and protein concentration in Bronchoalveolar lavage fluid (BALF) were dramatically decreased by ALA pretreatment. Treatment with ALA significantly alleviated the infiltration of total cells and neutrophils, while increased the number of the macrophages. ALA significantly inhibited the secretion of proinflammatory cytokines including tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and interleukin-1β (IL-1β) and increased anti-inflammatory cytokine. Moreover, we found that the levels of myeloperoxidase (MPO) and malondialdehyde (MDA) were highly increased in LPS-induced ALI, while the activities of glutathione (GSH) and superoxide dismutase (SOD) were decreased, which were reversed by ALA. ALA attenuated LPS-induced histopathological changes and apoptosis. Furthermore, ALA significantly inhibited the phosphorylation of IκBα and NF-κB (p65) activation in ALI. ALA showed anti-inflammatory effects in mice with LPS-induced ALI. NF-κB pathway may be involved in ALA mediated protective effects.

To make recommendations on the design and analysis of Piga gene mutation assay on the basis of the historical control data.All negative historical control data( total: 128) were collected to create the historical control data base, and descriptive statistics was used to determine the distribution of the data. The power of the number of animals per group, number of measured cells per animal and number of groups were discussed through simulation test based on historical data base.The group mean of mutant red blood cells( RBCs~(CD59-)) and mutant reticulocytes ( RETs~(CD59-)) were 26. 63 × 10~(-6) and 35. 85 × 10~(-6), respectively, and the standard deviation were 27. 71 × 10~(-6) and 31. 06 × 10~(-6), respectively. By mapping the frequencies of data, the variables had skewed distributions and translated to normal distributions after logarithmic transformation. The confidence level for population means were 100% and 92% for RBCs( 1 × 10~6 cells per animal) and RETs( 0. 3 × 10~6 cells per animal), and it increased to 100% when 1 × 10~6 cells was scored for RETs. Group sizes of 5 had low statistical power while the minimal detectable difference was 2 times. The power had increased to80% when 4- or 5- fold of minimal detectable difference was employed.In brief, the recommendations include:① A log( 10) transformation of mutant frequencies often has been found satisfactory for data when parametric method such as an analysis of variance are used. ② Young adult animals( approximately 6 weeks of age rats) are recommended in this assay. ③ The recommended number of RETs and RBCs are1 × 10~6 and5 × 10~7, respectively. ④ Based on power calculations, 3 treatment groups and 1 negative control group are appropriate when fourfold increase was employed. ⑤ Group sizes of 6 or 7 are recommended but 5 analyzable animals per group may be acceptable when 4 test groups were used.

Porcine circovirus type 2 (PCV2) has been identified as the etiologic agent which causing postweaning multisystemic wasting syndrome in swine farms in the world. Some quantitative proteomic studies showed that many proteins significantly changed in PCV2-infected cells. To explore the role of cellular chaperones during PCV2 infection, cytoprotective chaperone Hsp27 was analyzed in PCV2-infected PK-15 cells in this study. The results showed that Hsp27 could up-regulate and accumulate in phosphorylated forms in the nuclear zone during PCV2 replication. Suppression of Hsp27 phosphorylation with specific chemical inhibitors or downregulation of all forms of Hsp27 via RNA interference significantly reduced the virus replication. Meanwhile, over-expression of Hsp27 enhanced PCV2 genome replication and virion production. It indicated that Hsp27 was required for PCV2 production in PK-15 cells culture. It should be helpful for understanding the mechanism of replication and pathogenesis of PCV2 and development of novel antiviral therapies in the future.