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Zinc oxide nanoparticles (ZnONPs) are widely utilized across various industries, raising concerns about their potential toxicity, especially in the respiratory system. This study explores the role of autophagy, regulated by microtubule-associated protein 1A/1B-light chain 3B (LC3B), in ZnONPs-induced toxicity using both in vivo (LC3B knockout mice) and in vitro (BEAS-2B cells) models. Our findings demonstrate that LC3B-regulated autophagy mitigates ZnONPs-induced epithelial cell dysfunction and acute lung injury. In the absence of LC3B, oxidative stress, inflammation, and intracellular zinc accumulation are exacerbated, resulting in mitochondrial dysfunction and epithelial cell death. In vitro, LC3B knockdown disrupted zinc ion transporter expression and impaired mitophagic flux in BEAS-2B cells. Treatment with zinc ion chelators alleviated these toxic effects, confirming that free zinc ions play a critical role in driving ZnONPs toxicity. These findings highlight that targeting autophagy and maintaining zinc homeostasis could offer therapeutic strategies to reduce ZnONPs-induced lung damage., (© The Author(s) 2024. Published by Oxford University Press on behalf of the Society of Toxicology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Aim: Regenerative endodontic procedures (REPs) are tissue engineering-based treatments aimed at regenerating the pulp-dentin complex. However, these procedures are not widely practiced or well-known among dentists due to a lack of training. This study aims to develop a practical hands-on training model for REPs to enhance students' theoretical knowledge and practical skills., Methodology: A cost-effective, easy-to-operate, and reusable training model for preclinical training of REPs was developed. A total of 125 undergraduates were divided into an experimental group (REPs model) and a control group (traditional teaching). The students' theoretical knowledge and operational skills were assessed through examinations and practical assessments. Student perceptions were assessed by questionnaires., Results: The experimental group achieved higher scores in theoretical knowledge and operational skills compared to the control group (p < .001). The process evaluation of the experimental group demonstrated a high level of proficiency in executing root canal disinfection steps, with pass rates ranging from 93.5% to 95.2%. Students in both groups perceived the model as valuable for improving critical thinking, stimulating interest, and enhancing their confidence and proficiency., Conclusion: The developed REPs training model is a practical and effective tool for teaching and training regenerative endodontics. It bridges the gap between preclinical and clinical education, enhancing students' confidence and clinical competence., (© 2024 British Endodontic Society. Published by John Wiley & Sons Ltd.)

Objective: This study aims to explore the time threshold for defining prolonged mechanical ventilation (PMV) in children, along with its risk factors and outcomes., Methods: A prospective cohort study was conducted on children aged 29 days-18 years, who were consecutively admitted to two Pediatric Intensive Care Units (PICUs) at Children's Hospital of Chongqing Medical University, from October 2020 to June 2021. The study included patients receiving mechanical ventilation (MV) for more than 2 days (each day >6 h). Participants were divided into five groups based on the duration of MV (2-7 days, 8-14 days, 15-21 days, 21-30 days, >30 days) to compare rates of extubation failure, all-cause mortality one month post-discharge, incidence of ventilator-associated pneumonia, tracheotomy rates, total hospital stay, PICU stay, and overall hospital costs. The most clinically and statistically significant outcome variables were selected. The Youden index was used to determine the MV duration with the most significant impact on overall outcomes, defining this as PMV. Baseline characteristics, treatment information, and outcomes were compared between PMV and non-PMV groups. Univariate and multivariate logistic regression analyses were used to identify risk factors for PMV occurrence., Results: A total of 382 subjects were included in the study. The distribution of children across the five MV duration groups was 44.2%, 27.7%, 10.7%, 8.9%, and 8.4% respectively. The rates of at least one extubation failure in each group were 5.9%, 10.4%, 41.5%, 41.2%, and 46.9% (p < .05). Statistically significant differences were observed among groups in terms of tracheotomy rates, all-cause mortality at 1 month postdischarge, median total hospital stay, median PICU stay, and hospital costs (p < .05). Defining PMV, the most appropriate time point calculated was 12.5 days, based on at least one extubation failure and/or death within 1 month postdischarge. Higher PIM-3 scores, weight for age <-2SD, admission for respiratory distress/insufficient ventilation and/or hemodynamic instability/shock/arrhythmia, noninvasive ventilation on the first day, and undergoing blood transfusion treatment were identified as risk factors for PMV (p < .05)., Conclusion: In children, MV for ≥13 days significantly increases mortality rates, extubation failure and tracheotomy rates, duration of PICU and total hospital stay and costs. We suggest defining PMV as MV ≥13 days, particularly for children undergoing MV for respiratory illnesses. This definition can assist clinicians in developing appropriate treatment strategies by focusing on risk factors and providing reliable prognostic consultation to patients' families., (© 2024 Wiley Periodicals LLC.)

The concentration of heavy metals (HMs) in aquaculture pond sediments significantly affects aquatic food safety and environmental quality. The contamination characteristics, drivers and potential sources of HMs in typical bulk freshwater aquaculture pond sediments in major provinces of China were systematically investigated using a variety of methods and models. Specifically, 130 surface sediment samples were collected from the study area, and the geoaccumulation index (I geo ) and potential ecological risk index (RI) were used to jointly evaluate the characteristics of the HMs. Spearman's correlation and redundancy analysis revealed the main drivers of the HMs. Additionally, the positive matrix factorization (PMF) model and absolute principal component score-multiple linear regression (APCS-MLR) model were used to identify the sources of HMs. The results revealed that the pond sediments were safe for fish culture in most of the study areas. Aquafeed protein content is an important driver of HM concentrations in sediments. The total organic carbon (TOC) content, percentage of clay particles, and pH of the aquaculture pond sediments determined the sediment HMs enrichment abilities as 13.6 %, 52 %, and 9.8 %, respectively. Cd, a significantly enriched pollutant, posed a greater ecological risk than the other five HMs (Cr, Cu, Zn, As, and Pb). Three sources of HMs were identified, including agricultural activity (e.g., aquafeeds, pesticides, and fertilizers), industrial production, and natural sources, with contributions of 44.29 %, 36.66 %, and 19.05 %, respectively. This study provides a scientific basis for minimizing the input and accumulation of HMs in freshwater aquaculture pond sediments, and this can provide insights into the prevention and control of the ecological risks posed by HMs., Competing Interests: Declaration of competing interest The authors declare that they have no competing financial interests or personal relationships that may have influenced the work reported in this study., (Copyright © 2024. Published by Elsevier B.V.)

Polycyclic aromatic hydrocarbons (PAHs) are a group of extremely carcinogenic organic pollutants. Our previous findings have demonstrated that plant roots actively take up PAHs through co-transport with H + ions. Auxin serves as a pivotal regulator of plant growth and development. However, it remains unclear whether the hormone can enhance the uptake of PAHs by plant roots. Hence, the wheat root exposed to PAHs with/without auxins was set to investigate how the auxin promotes the PAHs uptake by roots. In our study, auxin could significantly enhance the uptake of PAHs after 4 h of exposure. After the addition of auxin, the root tissue cytoplasmic pH value was decreased and the H + influx was observed, indicating that the extracellular space was alkalinized in a short time. The increased H + influx rate enhanced the uptake of PAHs. In addition, the H + -ATPase activity was also increased, suggesting that auxin activated two distinct and antagonistic H + flux pathways, and the H + influx pathway was dominant. Our findings offer important information for exploring the mechanism underlying auxin regulation of PAHs uptake and the phytoremediation of PAH-contaminated soil and water., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Polycyclic aromatic hydrocarbons (PAHs) are a category of organic pollutants known for their high carcinogenicity. Our previous research has illustrated that plant roots actively absorb PAHs through a co-transport mechanism with H + ions. Because auxin can increase the H + -ATPase activity, the wheat roots were exposed to PAHs with/without auxins to study whether auxins facilitate the uptake of PAHs by plant roots and to gain insights into the underlying mechanisms of this process. In our study, indole acetic acid (100 μM) and α-naphthaleneacetic acid (10 μM) significantly increased the PAHs concentrations in apoplast and symplast, and the treating time and concentrations were positively correlated with PAHs accumulations. The time-dependent kinetics for 36 h followed the Elovich equation, and the concentration-dependent kinetics of apoplastic and symplastic uptake for 4 h could be described with the Freundlich and Michaelis-Menten equations, respectively. The proportion of PAHs accumulated in apoplast could be enhanced by auxins in most treatments. Our findings offer novel insights into the mechanisms of PAH uptake by plant roots under auxin exposure. Additionally, this research aids in refining strategies for ensuring crop safety and improving phytoremediation of PAH-contaminated soil and water., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

This work firstly reported a new polycaprolactone based material functionalized with guanidinium ionic liquid (PCL-GIL) as the stationary phase with high resolution performance for capillary gas chromatography (GC). It is composed of polycaprolactone (PCL) and guanidinium ionic liquid (GIL) with amphiphilic conformation. The PCL-GIL capillary column coated by static method exhibited high column efficiency of 3942 plates/m and moderate polarity. As a result, the PCL-GIL column exhibited high-resolution capability. For a mixture of 27 analytes with a wide ranging polarity and outperformed the PCL-2OH and HP-35 columns, showing its advantageous separation capability for analytes of diverse types. Moreover, the PCL-GIL column showed high resolving capability for various positional isomers and cis-/trans-isomers, including alkylbenzenes, chlorobenzenes, naphthalenes, bromonitrobenzenes, chloronitrobenzenes, benzaldehydes, phenols, alcohols, respectively. In a word, PCL derivatized by GIL units as a new type of stationary phase has a promising future in GC separations., (© 2023 Wiley-VHCA AG, Zurich, Switzerland.)

In this paper, an amphiphilic calix[8]arene with polyethylene glycol unit branches (C8A-PEG) was synthesized and applied for capillary gas chromatography (GC). The C8A-PEG was coated on the inner wall of a capillary column by a static method with the column efficiency of 3165 plates/m and polar nature. As demonstrated, the C8A-PEG column has excellent physicochemical properties and separation performance since it has π-electron-rich 3D cavity which combines with polar PEG units. Compared with two columns corresponding to the construction units C8A and PEG, the C8A-PEG column shows distinctly advantageous performance for the mixture of 22 components with diverse types. Impressively, it shows satisfactory resolution for positional isomers and cis-/trans- isomers, especially the challenging isomers of toluidine and dimethylaniline. The outstanding distinguishing capability of the C8A-PEG stationary phase is mainly attributed to the abundant molecular recognition interactions, including van der Waals, dipole-dipole, H-bonding and π-π stacking interactions. This work has proved that the new GC stationary phases constructed by different units can complement each other's advantages, improve their physicochemical properties and separation performance, and have broad application prospects in chromatographic analysis., (© 2023. The Author(s), under exclusive licence to The Japan Society for Analytical Chemistry.)

The separation of aromatic isomers, in particular xylene isomers, represents a big issue in chemical and petroleum industries, owing to their similar molecular sizes and boiling points. In this work, the investigation ofpillar[6]arene derivative modified by long alkyl chains (P6A-C10) as a stationary phase for high-resolution gas chromatographic (GC) separations of xylene isomers is presented. Pillar[n]arenes are a new class of macrocyclic hosts that can accommodate specific guests due to their highly symmetrical and rigid pillar architectures with π-electron rich cavities. The P6A-C10 column showed high-resolution performance towards xylene isomers, with peculiar advantages if compared with the commercial HP-5, HP-35, DB-17, and PEG-20Mcolumns.A quantum chemistry calculation has been performed, showing a difference in non-covalent interactions with the P6A-C10 pillar framework, which leads to specific selectivity for xylene isomers.Furthermore, the P6A-C10 column exhibited good repeatability., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Objective: The study was conducted to screen differentially expressed long noncoding RNA (lncRNA) in chickens by high-throughput sequencing and explore its mechanism of action on intramuscular fat deposition., Methods: Herein, Rose crown and Cbb broiler chicken embryo breast and leg muscle lncRNA and mRNA expression profiles were constructed by RNA sequencing. A total of 96 and 42 differentially expressed lncRNAs were obtained in Rose crown vs Cobb broiler chicken breast and leg muscle, respectively. lncRNA-ENSGALT00000046546, with high interspecific variability and a potential regulatory role in lipid metabolism, and its predicted downstream target gene 1-acylglycerol-3-phosphate-O-acyltransferase 2 (AGPAT2), were selected for further study on the preadipocytes., Results: lncRNA-46546 overexpression in chicken preadipocyte 2 cells significantly increased (p<0.01) the expression levels of AGPAT2 and its downstream genes diacylglycerol acyltransferase 1 and diacylglycerol acyltransferase 2 and those of the fat metabolism-related genes peroxisome proliferator-activated receptor γ, CCAAT/enhancer binding protein α, fatty acid synthase, sterol regulatory element-binding transcription factor 1, and fatty acid binding protein 4. The lipid droplet concentration was higher in the overexpression group than in the control cells, and the triglyceride content in cells and medium was also significantly increased (p<0.01)., Conclusion: This study preliminarily concludes that lncRNA-46546 may promote intramuscular fat deposition in chickens, laying a foundation for the study of lncRNAs in chicken early embryonic development and fat deposition.

Background: Studies in France, Korea, and Singapore found that G1-G6 transcriptomes are involved in hepatocellular carcinoma (HCC) carcinogenesis. However, the suitability of this method in Chinese HCC patients has remained unknown., Methods: The correlation between the G1-G6 molecular classification and clinicopathological features were analyzed in 107 Chinese HCC patients through the retrospective cohort study. RNA sequencing and bioinformatics analysis were performed to screen related targets and molecular signaling pathways., Results: We found that the G1-G3 subgroups were associated with high serum alpha-fetoprotein (AFP) level, high copy number of hepatitis B virus (HBV) DNA, complex histopathological structure, macrovascular invasion. The G1 subgroup was mainly related to liver cancer stemness, and G3 subgroup showed the worst prognosis. The G5 and G6 subgroups were associated with activation of the Wnt/β-catenin pathway. Compared with the G4-G6 group, the G1-G3 group showed significantly higher expression levels of regenerating family member 1 beta ( REG1B ), regenerating family member 3 gamma ( REG3G ), and inositol 1,4,5-trisphosphate receptor type 1 ( ITPR1 ), and enriched calcium signaling pathway., Conclusions: This study enhances our understanding of the heterogenicity of China HCC and indicates that the G1-G6 signatures can be used to identify predictive biomarkers against HCC patients in China.

In this work, the fabrication, synthesis, and characterization of a new stationary phase based on an amphiphilic pillar[6]arene (P6A-C10-2NH 2 ) for gas chromatographic analyses are reported. The gas chromatography (GC) column prepared with P6A-C10-2NH 2 stationary phase exhibited a medium polarity, an efficiency of 3219 plates/m, and unmatched resolving capabilities toward chloroaniline, bromoaniline, iodoaniline, and toluidine isomers. Furthermore, the P6A-C10-2NH 2 column showed excellent repeatability with maximum relative standard deviations equal to 0.02, 0.07, and 2.56% for run-to-run, day-to-day, and column-to-column, respectively, demonstrating a great potential as a new stationary phase in separation science.

Solar-induced fluorescence (SIF) is a promising proxy for photosynthesis, but it is unclear whether it performs well in tracking the gross primary productivity (GPP) under different environmental conditions. In this study, we investigated the dynamics of the two parameters from October 2020 to June 2021 in field-grown winter wheat (Triticum aestivum) and found that the ability of SIF to track GPP was weakened at low temperatures. Accounting for the coupling of light and temperature at a seasonal scale, we found that SIF yield showed a lower temperature sensitivity and had a lower but broader optimal temperature range compared with light-use efficiency (LUE), although both SIF yield and LUE decreased in low-temperature conditions. The discrepancy between the temperature responses of SIF yield and GPP caused an increase in the ratio of SIF/GPP in winter, which indicated the variation in the relationship between them during this period. The results of our study highlight the impact of low temperature on the relationship between SIF and GPP and show the necessity of reconsidering the dynamics of energy distribution inside plants under changing environments., (© The Author(s) 2022. Published by Oxford University Press on behalf of the Society for Experimental Biology. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

In this work, a novel capillary column (C4A-mPEG) with a calixarene-based polymer stationary phase (poly(ethylene glycol) methyl ether-functionalized 4-tert-butylcalix[4]arene) was designed and used for gas chromatographic (GC) separations. The C4A-mPEG capillary column, prepared by the static coating method, showed moderate polarity and a column efficiency of 2332 plates/m, determined by 1-octanol at 120 °C. The separation features of C4A-mPEG stationary phase, resulting from its unique structure and multiple molecular recognition processes with analytes, including π-π, H-bonding, dipole-dipole, and van der Waals interactions, allowed to obtain high-resolution performances for a wide range of compounds and their isomers, especially benzaldehydes, phenols, and anilines. Moreover, compared with 4-tertbutyl calix[4]arene (C4A) and polyethylene glycol (PEG) stationary phases, a higher resolving capability was also observed for the separation of toluidine and xylidine isomers., (© 2022 Wiley-VHCA AG, Zurich, Switzerland.)

Polycyclic aromatic hydrocarbons (PAHs) have the potential to cause cancer, teratogenicity, and mutagenesis in humans. Long-term plant safe production relies on how PAHs are transported and coordinated across organs. However, the acropetal transfer mechanism of PAHs in staple crop stems, particularly in xylem, a critical path, is unknown. Herein, we first confirmed the presence of specific interaction between the proteins and phenanthrene by employing the magnetic phenanthrene-bound bead immunoassay and label free liquid chromatograph mass spectrometer (LC-MS/MS), suggesting that peroxidase (uniprot accession: A0A3B5XXD0) and unidentified proteins (uniprot accession: A0A3B6LUC6) may function as the carriers to load and acropetally translocate phenanthrene (a model PAH) in wheat xylem. This specified binding of protein-phenanthrene may form through hydrophobic interactions in the conservative binding region, as revealed by protein structural investigations and molecular docking. To further investigate the role of these proteins in phenanthrene solubilization, phenanthrene exposure was conducted: a substantial quantity of peroxidase was produced; an unusually high expression of uncharacterized proteins was observed, indicating their positive effects in the acropetal transfer of phenanthrene in wheat xylem. These data confirmed that the two proteins are crucial in the solubilization of phenanthrene in wheat xylem sap. Our findings provide fresh light on the molecular mechanism of PAH loading in plant xylem and techniques for ensuring the security of staple crops and improving the efficacy of phytoremediation in a PAH-contaminated environment., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Response to cancer immunotherapy in primary versus metastatic disease has not been well-studied. We found primary pancreatic ductal adenocarcinoma (PDA) is responsive to diverse immunotherapies whereas liver metastases are resistant. We discovered divergent immune landscapes in each compartment. Compared to primary tumor, liver metastases in both mice and humans are infiltrated by highly anergic T cells and MHCII lo IL10 + macrophages that are unable to present tumor-antigen. Moreover, a distinctive population of CD24 + CD44 - CD40 - B cells dominate liver metastases. These B cells are recruited to the metastatic milieu by Muc1 hi IL18 hi tumor cells, which are enriched >10-fold in liver metastases. Recruited B cells drive macrophage-mediated adaptive immune-tolerance via CD200 and BTLA. Depleting B cells or targeting CD200/BTLA enhanced macrophage and T-cell immunogenicity and enabled immunotherapeutic efficacy of liver metastases. Our data detail the mechanistic underpinnings for compartment-specific immunotherapy-responsiveness and suggest that primary PDA models are poor surrogates for evaluating immunity in advanced disease., (© 2022. The Author(s), under exclusive licence to Springer Nature Limited.)

Pillar[n]arenes possess highly symmetrical and rigid pillar-shaped architecture with π-electron rich cavity that afford their reliable host-guest recognition interactions towards matched guests. In this work, a novel amphiphilic pillar[5]arene (P5A-C10-2NH 2 ) was designed, synthesized and employed as the stationary phase for capillary gas chromatography. To date, they have not been reported in the field of chromatography. The P5A-C10-2NH 2 capillary column (10 m × 0.25 mm i.d.) was prepared by static coating method. Its capillary column exhibited moderate polarity and column efficiency of 2265 plates/m determined by naphthalene at 120 °C. As evidenced, the P5A-C10-2NH 2 column achieved advantageous separation performance for a mixture of 24 analytes of diverse types and exhibited different chromatographic selectivity from two pillar[5]arene derivatives columns and commercial HP-35 column with 35%-phenyl-methylpolysiloxane. Moreover, the P5A-C10-2NH 2 column baseline resolved more than a dozen positional and cis-trans isomers. Furthermore, the separation mechanism of P5A-C10-2NH 2 column was discussed by quantum chemical calculations. In addition, the P5A-C10-2NH 2 column had high thermal stability and excellent separation repeatability 0.01-0.04% for run-to-run, 0.03-0.17% for day-to-day and 3.2-3.9% for column-to-column. The special amphiphilic structure and high resolution for various analytes reveal the good potential of pillararenes as a new class of stationary phases for chromatographic analyses. Moreover, the TPG column achieved improved thermal stability over the GIL column and excellent repeatability., Competing Interests: Declaration of Competing Interest Authors declare no conflict of interest., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Nanoscale plastic particles are widely found in the terrestrial environment and being increasingly studied in recent years. However, the knowledge of their translocation and accumulation mechanism controlled by nanoplastic characterizations in plant tissues is limited, especially in plant cells. Here, 20 mg L -1 polystyrene nanoparticles (PS NPs) with different sizes and amino/carboxy groups were employed to investigate the internalization process in wheat roots and cells. From the results, we found that the uptake of small-size PS NPs in the root tissues was increased compared to that of large-size ones, but no PS NPs were observed in the vascular cylinder. Similar results were observed in their cellular uptake process. Besides, the cell wall could block the entry of large-size PS NPs while the cell membrane could not. The -NH 2 group on the PS NPs surface could benefit their tissular/cellular translocation compared to the -COOH group. The internalization of PS NPs was controlled by both particle size and surface functional group, and the size should be the primary factor. Our findings offer important information for understanding the PS NPs behaviors in plant tissues, especially at the cellular level, and assessing their potential risk to food safety, quality, and agricultural sustainability.

The problem of algal bloom caused by eutrophication has attracted global attention. Many scholars have studied the problem associated with algae bloom, but few have carried out dynamic monitoring, instead focusing on the formation mechanism of cyanobacteria. For our study of the Taihu Lake in China, we used Moderate-Resolution Imaging Spectroradiometer (MODIS) and Landsat remote sensing image data from 2017 to establish a prediction model. First, we used MODIS data to retrieve the concentration of N, P, and chlorophyll a in water. Then, we applied the analytic hierarchy process (AHP) model to the inversion results to construct the diffusion potential index. Finally, we used C# to compile the cellular automata (CA) model. We found that the distribution of cyanobacteria predicted by our method was consistent with the algal bloom situation of Taihu Lake in 2017. The results showed that the method effectively predicts the dynamic transfer of cyanobacteria from outbreak to diffusion in a short period of time, which can help decision-makers monitor lake health., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

The gut microbiome shapes local and systemic immunity. The liver is presumed to be a protected sterile site. As such, a hepatic microbiome has not been examined. Here, we showed a liver microbiome in mice and humans that is distinct from that of the gut and is enriched in Proteobacteria. It undergoes dynamic alterations with age and is influenced by the environment and host physiology. Fecal microbial transfer experiments revealed that the liver microbiome is populated from the gut in a highly selective manner. Hepatic immunity is dependent on the microbiome, specifically the bacteroidetes species. Targeting bacteroidetes with oral antibiotics reduced hepatic immune cells by approximately 90%, prevented antigen-presenting cell (APC) maturation, and mitigated adaptive immunity. Mechanistically, our findings are consistent with presentation of bacteroidetes-derived glycosphingolipids to NKT cells promoting CCL5 signaling, which drives hepatic leukocyte expansion and activation, among other possible host-microbe interactions. Collectively, we reveal a microbial/glycosphingolipid/NKT/CCL5 axis that underlies hepatic immunity.

Health risks of typical benzene series and halocarbons (BSHs) in a densely populated area near a large-scale chemical industrial park were investigated. Ambient and indoor air and tap water samples were collected in summer and winter; and the concentration characteristics, sources, and exposure risks of typical BSH species, including five benzene series (benzene, toluene, ethylbenzene, o-xylene, m,p-xylene) and five halocarbons (dichloromethane, trichloromethane, trichloroethylene, tetrachloromethane, and tetrachloroethylene), were analysed. The total mean concentrations of BSHs were 53.32 μg m -3 , 36.29 μg m -3 , and 26.88 μg L -1 in indoor air, ambient air, and tap water, respectively. Halocarbons dominated the total BSHs with concentrations relatively higher than those in many other industrial areas. Industrial solvent use, industrial processes, and vehicle exhaust emissions were the principal sources of BSHs in ambient air. The use of household products (e.g., detergents and pesticides) was the principal source of indoor BSHs. Inhalation is the primary human exposure route. Ingestion of drinking water was also an important exposure route but had less impact than inhalation. Lifetime non-cancer risks of individual and cumulative BSHs were below the threshold (HQ = 1), indicating no significant lifetime non-cancer risks in the study area. However, tetrachloromethane, benzene, trichloromethane, ethylbenzene, and trichloroethylene showed potential lifetime cancer risk. The cumulative lifetime cancer risks exceeded the tolerable benchmark (1 × 10 -4 ), indicating a lifetime cancer risk of BSHs to residents near the chemical industry park. This study provides valuable information for the management of public health in chemical industrial parks., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

1. Si-Ni-San (SNS) possesses extensive therapeutic effects, however, the extent to which main components are absorbed and the mechanisms involved are controversial. 2. In this study, MDCK cell model was used to determine the permeability characteristics and interaction between the major components of Si-Ni-San, including saikosaponin a, paeoniflorin, naringin and glycyrrhizic acid. 3. The transport of the major components was concentration-dependent in both directions. Moreover, the transport of paeoniflorin, naringin and glycyrrhizic acid was significantly reduced at 4 °C or in the presence of NaN3. Additionally, the efflux of paeoniflorin and naringin were apparently reduced in the presence of P-gp inhibitor verapamil. The transport of glycyrrhizic acid was clearly inhibited by the inhibitors of MRP2, indicating that MRP2 may be involved in the transport of glycyrrhizic acid. However, the results indicated that saikosaponin a was absorbed mainly by passive diffusion. Furthermore, the combined incubation of four major components had a powerful sorbefacient effect than a single drug used alone which may be regulated by tight junctions. 4. Taken together, our study provides useful information for pharmacological applications of Si-Ni-San and offers new insights into this ancient decoction for further researches, especially in drug synergism.

Piezo1 is a mechanosensitive ion channel that has gained recognition for its role in regulating diverse physiological processes. However, the influence of Piezo1 in inflammatory disease, including infection and tumor immunity, is not well studied. We postulated that Piezo1 links physical forces to immune regulation in myeloid cells. We found signal transduction via Piezo1 in myeloid cells and established this channel as the primary sensor of mechanical stress in these cells. Global inhibition of Piezo1 with a peptide inhibitor was protective against both cancer and septic shock and resulted in a diminution in suppressive myeloid cells. Moreover, deletion of Piezo1 in myeloid cells protected against cancer and increased survival in polymicrobial sepsis. Mechanistically, we show that mechanical stimulation promotes Piezo1-dependent myeloid cell expansion by suppressing the retinoblastoma gene Rb1 We further show that Piezo1-mediated silencing of Rb1 is regulated via up-regulation of histone deacetylase 2. Collectively, our work uncovers Piezo1 as a targetable immune checkpoint that drives immunosuppressive myelopoiesis in cancer and infectious disease., (Copyright © 2020 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works.)

Programmed cell death protein 1 (PD-1) ligation delimits immunogenic responses in T cells. However, the consequences of programmed cell death 1 ligand 1 (PD-L1) ligation in T cells are uncertain. We found that T cell expression of PD-L1 in cancer was regulated by tumor antigen and sterile inflammatory cues. PD-L1 + T cells exerted tumor-promoting tolerance via three distinct mechanisms: (1) binding of PD-L1 induced STAT3-dependent 'back-signaling' in CD4 + T cells, which prevented activation, reduced T H 1-polarization and directed T H 17-differentiation. PD-L1 signaling also induced an anergic T-bet - IFN-γ - phenotype in CD8 + T cells and was equally suppressive compared to PD-1 signaling; (2) PD-L1 + T cells restrained effector T cells via the canonical PD-L1-PD-1 axis and were sufficient to accelerate tumorigenesis, even in the absence of endogenous PD-L1; (3) PD-L1 + T cells engaged PD-1 + macrophages, inducing an alternative M2-like program, which had crippling effects on adaptive antitumor immunity. Collectively, we demonstrate that PD-L1 + T cells have diverse tolerogenic effects on tumor immunity.

A facile one-pot solvent thermal method was proposed to synthesize magnetic mesoporous carbon (MMC) using Fe(NO 3 ) 3 ·9H 2 O as a precursor, Pluronic copolymer P123 as template, and chitosan as carbon source, and it was applied for the adsorptive remediation of methyl orange (MO). The characterization results of TEM, XRD, and IR showed that MMC consisted of graphitized carbon matrix and some black spherical particle mixture of Fe 3 O 4 and Fe, and it was rich in hydroxyl and carbonyl groups. Besides, the effect of the content of Fe and the content of chitosan in MMC on the magnetism and adsorption performance of prepared material were investigated. In addition, the effects of pH value, initial concentration of methyl orange, and contact time on the adsorption performance of MO were studied, respectively. At 318 K, the maximum adsorption capacity of MO calculated from Langmuir isotherm was from 139 to 400 mg g -1 on MMC. Kinetic studies demonstrated that the adsorption process obeyed a pseudo-second-order kinetic model. The regeneration experiments revealed that MMC could be reused at least five times without notable decrease of adsorption performance. These results illustrate that MMC is an efficient and economical adsorbent for the adsorption of MO.

Although CD4 + FOXP3 + T regulatory (Treg) cells are well-known mediators of immunologic tolerance, their influences in the tumor microenviroment are incompletely understood. Writing in this issue of Cancer Discovery , Zhang and colleagues demonstrate that in pancreatic cancer, Treg cells promote the differentiation of tumor-restraining myofibroblastic cancer-associated fibroblasts, challenging the existing notion that Treg cells enable tumor progression. See related article by Zhang et al., p. 422 ., (©2020 American Association for Cancer Research.)

Background and Aims: The recruitment and activation of inflammatory cells in the liver delineates the transition from hepatic steatosis to steatohepatitis (SH)., Approach and Results: We found that in SH, γδT cells are recruited to the liver by C-C chemokine receptor (CCR) 2, CCR5, and nucleotide-binding oligomerization domain-containing protein 2 signaling and are skewed toward an interleukin (IL)-17A + phenotype in an inducible costimulator (ICOS)/ICOS ligand-dependent manner. γδT cells exhibit a distinct Vγ4 + , PD1 + , Ly6C + CD44 + phenotype in SH. Moreover, γδT cells up-regulate both CD1d, which is necessary for lipid-based antigens presentation, and the free fatty acid receptor, CD36. γδT cells are stimulated to express IL-17A by palmitic acid and CD1d ligation. Deletion, depletion, and targeted interruption of γδT cell recruitment protects against diet-induced SH and accelerates disease resolution., Conclusions: We demonstrate that hepatic γδT cells exacerbate SH, independent of IL-17 expression, by mitigating conventional CD4 + T-cell expansion and modulating their inflammatory program by CD1d-dependent vascular endothelial growth factor expression., (© 2019 by the American Association for the Study of Liver Diseases.)

Pigs are animals that live in groups and have social hierarchies within the group. After mixing, they can re-establish social hierarchies within several days through fighting. Dominance hierarchical indices, such as I&SI, Elo rating, and Glicko rating, have been used to analyze social hierarchies of some social animals but not pigs. I&SI index involves iterative calculations that first minimize the number of inconsistencies (I) in a dominance matrix, and then minimize the strength of those inconsistencies (SI). Elo rating and Glicko rating indices are based on the sequence in which interactions occur, and continuously update ratings by looking at interactions sequentially. To study the temporal dynamics of social hierarchy formation and maintenance in weaned pigs after mixing, a total of 102 pigs (47 females and 55 barrows) were selected by similar body weight and mixed in 10 pens (10 or 11 females or barrows per pen). Their behavior was recorded and observed for 72 h after mixing. Results showed that hierarchical indices I&SI, Elo rating, and Glicko rating were associated with each other (| r | = 0.681 ~ 0.942, p < 0.001). I&SI was associated with logarithms of frequency of active attack (| r | = 0.65, p < 0.05) and tended to associated with logarithms of frequency of standoff (| r | = 0.48, p < 0.1). Elo rating, and Glicko rating were associated with the logarithms of duration of being bullied (| r | = 0.393~0.401, p < 0.05). In addition, Glicko rating tended to be associated with the logarithms of duration of active attack and frequency of active attack (| r | = 0.416~0.439, p < 0.1). Multiple linear regression analyses of logarithms of dyadic behavior indicators for three hierarchical indices indicated the same effects. The time (hours) to achieve social stability of pigs after mixing was lower for females than barrows (23.06 ± 4.15 vs. 40.55 ± 4.71 h; p < 0.001). The most dominant pig (the first ranked) in each pen quickly appeared within a few hours after mixing and remained stable. Overall, our study demonstrated that the ranks calculated by the three dominance hierarchical indices: I&SI, Elo rating, and Glicko rating, were consistent and partially associated with part of the dyadic behavioral indicators in weaned pigs after mixing.

Via promoting synovitis, pannus growth and cartilage/bone destruction, fibroblast-like synovial cells (FLSs) play a significant role in the pathogenesis of rheumatoid arthritis (RA). In our study, rats were induced with complete freund's adjuvant (CFA) to be animal models for studying the RA pathogenesis. Microtubule-associated Serine/Threonine-protein kinase 3 (MAST3) has been documented to play a critical role in regulating the immune response of IBD (Inflammatory bowel disease) and involved in the process of cytoskeleton organization, intracellular signal transduction and peptidyl-serine phosphorylation, but its role in the progression of RA remains unknown and is warranted for investigation. So, we tried our best to investigate the mechanism and signaling pathway of MAST3 in RA progression. In the synovial tissue and FLSs of AA rats, we have found that MAST3 was significantly up-regulated than normal. Furthermore, MAST3 overexpression could promote proliferation and inflammatory response of FLSs. In the aspect of mechanism, we discovered that the expression of MAST3 might involve in NF-κB signaling pathway in RA. On the whole, our results suggested that MAST3 might promote the proliferation and inflammation of FLSs by regulating NF-κB signaling pathway., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Understanding the genetic background underlying the expression of behavioral traits has the potential to fasten the genetic progress for reduced aggressive behavior of pigs. The monoamine oxidase A ( MAOA ) gene is known as the "warrior" gene, as it has been previously linked to aggressive behavior in humans and livestock animals. To identify single nucleotide polymorphisms in porcine MAOA gene associated with aggressive behavior of pigs, a total of 500 weaned pigs were selected and mixed in 51 pens. In each pen, two aggressive and two docile pigs (a total of 204 pigs) were selected based on their composite aggressive score (CAS). Ear tissue was sampled to extract genomic DNA. Constructs containing variable lengths of truncated porcine MAOA promoter were used to determine the promoter activity by a dual luciferase reporter system. The core promoter region was located at -679 bp to -400 bp. A total of nine single nucleotide polymorphisms (SNPs) in MAOA gene were genotyped, of which six SNPs had significant differences ( p < 0.05) in allele frequency between the aggressive and docile pigs. Linkage disequilibrium and association analyses showed that the pigs inherited the wild genotypes showed more aggressive behavior ( p < 0.05) than pigs with the mutant genotypes of the four linked SNPs, rs321936011, rs331624976, rs346245147, and rs346324437. In addition, pigs of GCAA haplotype were more ( p < 0.05) aggressive than the pigs with GCGA or ATGG haplotype. The construct containing the wild genotype GG of rs321936011 had lower ( p = 0.031) promoter activity compared to the mutant genotype AA. These results suggest that the four linked SNPs in MAOA gene could be considered as a molecular marker for behavioral trait selection in pigs.

Bacterial dysbiosis accompanies carcinogenesis in malignancies such as colon and liver cancer, and has recently been implicated in the pathogenesis of pancreatic ductal adenocarcinoma (PDA) 1 . However, the mycobiome has not been clearly implicated in tumorigenesis. Here we show that fungi migrate from the gut lumen to the pancreas, and that this is implicated in the pathogenesis of PDA. PDA tumours in humans and mouse models of this cancer displayed an increase in fungi of about 3,000-fold compared to normal pancreatic tissue. The composition of the mycobiome of PDA tumours was distinct from that of the gut or normal pancreas on the basis of alpha- and beta-diversity indices. Specifically, the fungal community that infiltrated PDA tumours was markedly enriched for Malassezia spp. in both mice and humans. Ablation of the mycobiome was protective against tumour growth in slowly progressive and invasive models of PDA, and repopulation with a Malassezia species-but not species in the genera Candida, Saccharomyces or Aspergillus-accelerated oncogenesis. We also discovered that ligation of mannose-binding lectin (MBL), which binds to glycans of the fungal wall to activate the complement cascade, was required for oncogenic progression, whereas deletion of MBL or C3 in the extratumoral compartment-or knockdown of C3aR in tumour cells-were both protective against tumour growth. In addition, reprogramming of the mycobiome did not alter the progression of PDA in Mbl- (also known as Mbl2) or C3-deficient mice. Collectively, our work shows that pathogenic fungi promote PDA by driving the complement cascade through the activation of MBL.

Kinetic studies of photo- and photoelectro-catalysis fixation of CO 2 are rare. Herein, a typical CO 2 reduction addition to trans-stilbene is studied. Through Tafel analyses, the reaction rate-determining step (RDS) is identified as the first step of an anion free radical generation from the substrate, and the reaction order is 0.5.

Long noncoding RNAs (lncRNAs) have been reported to play diverse roles in biologic and pathologic processes, including myogenesis. We found that lncRNA AK017368 is highly expressed in skeletal muscle cells. Functional analyses showed that overexpression of AK017368 promoted proliferation and restrained differentiation of myoblasts; whereas inhibition of AK017368 had completely opposite effects in vitro In mice, knockdown of AK017368 promoted muscle hypertrophy in vivo RNA molecules of AK017368 acted mechanistically as competing endogenous RNAs to target micro-RNA (miR)-30c, which was supported by the results of bioinformatics analyses and dual-luciferase reporter assays. It has been shown that lncRNA AK017368 competes with trinucleotide repeat containing-6A ( Tnrc6a ) for miR-30c. Tnrc6a was previously reported to promote proliferation and inhibit differentiation of myoblast cells, whereas miR-30c targets the 3'-UTR of Tnrc6a mRNA to weaken its function. Taken together, lncRNA AK017368 promotes proliferation and inhibits differentiation of myoblast cells by attenuating function of miR-30c.-Liang, T., Zhou, B., Shi, L., Wang, H., Chu, Q., Xu, F., Li, Y., Chen, R., Shen, C., Schinckel, A. P. lncRNA AK017368 promotes proliferation and suppresses differentiation of myoblasts in skeletal muscle development by attenuating the function of miR-30c., (© FASEB.)

Due to the decrease of intracellular NADH availability, gluconate metabolism is more conducive to pyruvate production than glucose. Transcriptome analysis revealed that the Entner-Doudoroff (ED) pathway was activated by gluconate in Escherichia coli YP211 (MG1655 ΔldhA ΔpflB Δpta-ackA ΔpoxB Δppc ΔfrdBC). To construct a new pyruvate producing strain with glucose metabolism via ED pathway, the genes ppsA, ptsG, pgi and gnd were deleted sequentially to reduce the demand for PEP and block the Embden-Meyerhor-Parnas pathway and Pentose-Phosphate pathway. After nearly 1000 generations of growth-based selection, the evolved strain YP404 was isolated and the ED pathway was proved to be activated as the primary glycolytic pathway. Comparing with YP211, the pyruvate concentration and yield increased by 59% and 10.1%, respectively. In fed-batch fermentation, the pyruvate concentration reached 83.5 g l -1 with a volumetric productivity of 2.3 g l -1  h -1 . This was the first time to produce pyruvate via ED pathway, and prove that this was a more effective way., (Copyright © 2017 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.)

1. Hesperetin (HDND) possesses extensive bioactivities, however, its poor solubility and low bioavailability limit its application. HDND-7, a derivative of HDND, has better solubility and high bioavailability. In this study, we investigated the intestinal absorption mechanisms of HDND-7. 2. MDCK cells were used to examine the transport mechanisms of HDND-7 in vitro, and a rat in situ intestinal perfusion model was used to characterize the absorption of HDND-7. The concentration of HDND-7 was determined by HPLC. 3. In MDCK cells, HDND-7 was effectively absorbed in a concentration-dependent manner in both directions. Moreover, HDND-7 showed pH-dependent and TEER-independent transport in both directions. The transport of HDND-7 was significantly reduced at 4 °C or in the presence of NaN3. Furthermore, the efflux of HDND-7 was apparently reduced in the presence of MRP2 inhibitors MK-571 or probenecid. However, P-gp inhibitor verapamil had no effect on the transport of HDND-7. The in situ intestinal perfusion study indicated HDND-7 was well-absorbed in four intestinal segments. Furthermore, MRP2 inhibitors may slightly increase the absorption of HDND-7 in jejunum. 4. In summary, all results indicated that HDND-7 might be absorbed mainly by passive diffusion via transcellular pathway, MRP2 but P-gp may participate in the efflux of HDND-7.

Estrus expression by gilts and sows is hereditable and important for heat detection. To better understand the molecular biological mechanisms of estrus expression in gilts, the mRNA expression profiles of follicular tissue from Large White gilts in diestrus (LD, n = 3) and estrus (LE, n = 3), and Chinese indigenous Mi gilts in diestrus (MD, n = 2) and estrus (ME, n = 3) were investigated using RNA sequencing. We detected 122,804-335,295 SNPs, 6,140-14,947 InDel and 12 types of AS events (39.57% TSS, 34.90% TTS) in 11 samples. A total of 2,838 differentially expressed genes (DEGs) were found in LD vs MD, LE vs ME, LE vs LD, or ME vs MD comparisons. Two DEGs (ACP5 and PIGS) were observed in all comparisons. Two new genes (ENSSSCG00000028235 and ENSSSCG00000021903) were exclusively expressed in Mi and Large White gilts, respectively. Bioinformatics analyses indicate that these DEGs are involved in single-organism process, catalytic activity, cell adhesion and enriched in ECM-receptor interaction, olfactory transduction, ovarian steroidogenesis, steroid biosynthesis and CAMs signaling pathways. These results of RNA-Seq have provided important information for screening the key functional genes or molecular markers of estrus expression in gilts.

Background: MTBH, a novel hesperetin derivative, possesses in vivo hepatoprotective effects against carbon tetrachloride (CCl 4 )-induced acute liver injury in Institute of Cancer Research (ICR) mice., Objectives: This study investigated the pharmacokinetics and tissue distribution of MTBH and its conjugated metabolites in rats after a single dose of MTBH., Methods: Male Sprague-Dawley (SD) rats were orally administered (25, 50, 100 mg/kg) or intravenously administered (25 mg/kg) MTBH and blood samples were withdrawn at specific times. Moreover, after a single oral dose of MTBH (200 mg/kg), tissues (heart, liver, spleen, lung, kidney, stomach, intestine, brain and muscle) were collected at scheduled time points., Results: The concentration of MTBH in plasma and tissues was assayed by HPLC before and after hydrolysis with β-glucuronidase or sulfatase. The glucuronides/sulfates were extensively present in the plasma, moreover, the free form was detectable in the plasma, but in a small amount equivalent to nearly 0.85-1.46 % of the amount of glucuronides/sulfates, the absolute bioavailability of MTBH was approximately 31.27 %. In tissues, the free form appeared in all tissues examined, with trace amount in brain and muscle, and considerable concentration in stomach and lung. Glucuronides/sulfates were the major forms in intestine, kidney and liver, whereas not detectable in heart, brain and muscle. The liver and intestine were found likely to accumulate MTBH at a high concentration among all tissues., Conclusions: The free form of MTBH was present in the circulation and all assayed organs, whereas its glucuronides/sulfates were the major forms in plasma and intestine, kidney and liver after a single dose.

Efficiency and fidelity are the key obstacles for genome editing toolboxes. In the present study, a PCR-based tandem repeat assisted genome editing (TRAGE) method with high efficiency and fidelity was developed. The design of TRAGE is based on the mechanism of repair of spontaneous double-strand breakage (DSB) via replication fork reactivation. First, cat-sacB cassette flanked by tandem repeat sequence was integrated into target site in chromosome assisted by Red enzymes. Then, for the excision of the cat-sacB cassette, only subculturing is needed. The developed method was successfully applied for seamlessly deleting, substituting and inserting targeted genes using PCR products. The effects of different manipulations including sucrose addition time, subculture times in LB with sucrose and stages of inoculation on the efficiency were investigated. With our recommended procedure, seamless excision of cat-sacB cassette can be realized in 48 h efficiently. We believe that the developed method has great potential for seamless genome editing in E. coli.

A sensitive and reliable HPLC-MS/MS method was developed and validated for the determination of free (unconjugated), glucuronidated, sulfated, and total (free and conjugated) HDND-7 in rat plasma and tissues. Plasma and tissues samples were treated prior to and after the enzyme hydrolysis. Chromatographic separation was achieved on a Phenomenex Luna C18 column (150 × 4.6mm, 3 μm), using isocratic mobile phase consisting of 0.1% formic acid-acetonitrile (50:50, v/v) at a flow rate of 300 μl/min. The detection was performed on a triple quadruple tandem mass spectrometer using positive electrospray ionization (ESI) source with a chromatographic run time of 5.0 min. The detection was operated by multiple reaction monitoring (MRM) of the transitions of m/z 429.3 → 223.9 for HDND-7 and 272.9 → 152.9 for naringenin (IS), respectively. This method was validated in terms of specificity, linearity, precision, accuracy, and stability. The calibration curves for plasma and tissues were linear over a wide concentration range of 0.02-40 μg/ml with a lower limit of quantification (LLOQ) of 0.02 μg/ml. Mean extraction recoveries in plasma and tissues ranged from 87.4 to 97.1% and from 54.2 to 70.5%, respectively. The intra- and inter-day precision values were below 15% and the accuracy was within ± 15%. The samples were stable under all the tested conditions. This method has been successfully applied to the pharmacokinetic study following oral doses of 25, 50 and 100mg/kg and intravenous dose of 25mg/kg, and tissue distribution study following oral dose of 50mg/kg., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Hesperetin, the aglycone of hesperidin, occurs naturally in citrus fruits. It exerts extensive pharmacological activities. However, hesperetin's poor solubility and low bioavailability limit its wide application. In order to overcome these limitations, recently a series of novel hesperitin derivatives containing Mannich base moieties were synthesized and the anti-inflammatory activity was evaluated, among which MTBH (8-methylene-tert-butylamine-3',5,7-trihydroxy-4'-methoxyflavanone) showed a significantly improved water solubility, and promising anti-inflammatory activity in vitro and in vivo compared with hesperitin. Thus, the aim of this study was to investigate the permeability and transport mechanisms of MTBH, using Caco-2 cell monolayer. MTBH was effectively absorbed by Caco-2 cells in a concentration-dependent manner in both directions at 7.5-480 μM. Moreover, MTBH showed pH dependent and TEER values independent transport in both directions. Transport of MTBH was obviously decreased in the presence of sodium azide (an ATP inhibitor) or CCCP (a proton-ionophore). MTBH transport was markedly reduced by MCT inhibitors quercetin or phloretin, and the substrate analogs l-lactate or benzoic acid. We verified MCT1, MCT3, MCT4, MCT5, and MCT6 were expressed in Caco-2 cells by western blot. Silence MCT1 with siRNA resulted in significant inhibition of MTBH uptake. The verapamil, a P-gp inhibitor, and Ko143, a BCRP inhibitor, had no effect on the transport of MTBH. However, MK-571 or probenecid, MRP2 inhibitors, led to an apparently decrease in the efflux of MTBH. In summary, MTBH was absorbed by transcellular passive diffusion and a pH dependent mechanism mediated by MCT1. MRP2 but P-gp or BCRP may be involved in the transport of MTBH., (Copyright © 2015 Elsevier B.V. All rights reserved.)

To develop convenient, effective cold chain-free subunit vaccines, a mannose-PEG-cholesterol conjugate (MPC) was synthesized as a lectin binding molecule and anchored onto liposomes which entrapped lipid A and model antigen to form a vaccine adjuvant-delivery system targeting antigen presenting cells. With MPC, soy phosphatidylcholine, stearylamine and monophosphoryl lipid A as emulsifiers dissolved in oil phase (O), and sucrose and BSA in water phase (W), the O/W emulsions were prepared and subsequently lyophilized. The lyophilized product was stable enough to be stored at room temperature and, upon rehydration, formed MPC-/lipid A-liposomes (MLLs) with a size under 300 nm and antigen association rates of around 36%. The MLLs given to mice via oral mucosal (o.m.) administration showed no side effects but induced potent immune responses as evidenced by the high levels of IgG in the sera and IgA in the salivary, intestinal and vaginal secretions of mice. High levels of IgG2a and IFN-γ in treated mice revealed that MLLs via o.m. vaccination induced a mixed Th1/Th2 response against antigens, establishing both humoral and cellular immunity. Thus, the MLLs may be a potent cold chain-free oral mucosal vaccine adjuvant-delivery system., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Using a procedure of emulsification-lyophilization (PEL), adjuvant lipid A-cochleates (LACs) were prepared as a carrier for model antigen bovine serum albumin (BSA). With phosphatidylserine and lipid A as emulsifiers dissolved in oil phase (O), sucrose and CaCl2 in the inner water phase (W1), and BSA, sucrose and PEG2000 in the outer water phase (W2), the W1/O/W2 emulsions were prepared and subsequently lyophilized to form a dry product which was stable enough to be stored at room temperature. Upon rehydration of the dry products, cochleates formed with a size of 800 nm and antigen association rates of 38%. After vaccination of mice through oral mucosal (o.m.) administration, LACs showed no side effects but induced potent immune responses as evidenced by high levels of IgG in the sera and IgA in the salivary, intestinal and vaginal secretions of mice. In addition, high levels of IgG2a and IFN-γ in the sera or culture supernatants of splenocytes of the immunized mice were also detected. These results revealed that LACs induced a mixed Th1/Th2 response against the loaded antigens. Thus, the LACs prepared by PEL were able to induce both systemic and mucosal immune responses and may act as a potent cold-chain-free oral mucosal vaccine adjuvant delivery system (VADS)., (Copyright © 2014 Elsevier B.V. All rights reserved.)