Water and ions transport in calcium silicate hydrate: a molecular dynamics study

Transport properties of water and ions in calcium silicate hydrate (C-S-H) greatly affect the durability of cementitious materials. In this study, molecular dynamics (MD) technique is used to investigate the transport behaviors of NaCl solution in C-S-H nanopores with different sizes (from 0.5 nm to 5 nm), and the hindering effect of C-S-H on the diffusion of water molecules and Cl ions is further explored in the case of a 5 nm pore. Results show that the diffusion coefficients of water molecules and Cl ions in C-S-H nanopores increase with the expansion of nanopore. At the atomic scale, the Ca-rich C-S-H forms Ca-O and Ca-Cl clusters with water molecules and Cl ions, respectively, and the Si-O tetrahedra on silicate chains can also build hydrogen bonding interactions with water molecules, which constrain the transport behaviors of water and ions. From the molecular perspective, this study innovatively investigates the effect of C-S-H pore size on the diffusion capacity of water and ions, and reveals the chemical bonding mechanism between water molecules, Cl ions and C-S-H, which provides a theoretical basis for studying the resistance of concrete to ionic attack

A compendium of genetic regulatory effects across pig tissues

The Farm Animal Genotype-Tissue Expression (FarmGTEx) project has been established to develop a public resource of genetic regulatory variants in livestock, which is essential for linking genetic polymorphisms to variation in phenotypes, helping fundamental biological discovery and exploitation in animal breeding and human biomedicine. Here we show results from the pilot phase of PigGTEx by processing 5,457 RNA-sequencing and 1,602 whole-genome sequencing samples passing quality control from pigs. We build a pig genotype imputation panel and associate millions of genetic variants with five types of transcriptomic phenotypes in 34 tissues. We evaluate tissue specificity of regulatory effects and elucidate molecular mechanisms of their action using multi-omics data. Leveraging this resource, we decipher regulatory mechanisms underlying 207 pig complex phenotypes and demonstrate the similarity of pigs to humans in gene expression and the genetic regulation behind complex phenotypes, supporting the importance of pigs as a human biomedical model.</p

Screening of <i>Codonopsis radix</i> Polysaccharides with Different Molecular Weights and Evaluation of Their Immunomodulatory Activity In Vitro and In Vivo

Polysaccharide is one of the main components of Codonopsis radix (CR) and has good immune activity. However, the immune activity of CR polysaccharides with different molecular weights has not been systematically screened. In this study, the polysaccharides of CR from Pingshun of Shanxi Province (PSDSs) were first divided into two groups using ultrafiltration: 3.3 kDa (PSDSs-1) and more than 2000 kDa (PSDSs-2). The immunomodulatory effects of PSDSs with different molecular weights were evaluated in vitro and in vivo. In vitro experimental results showed that compared with Lipopolysaccharide-induced macrophages, PSDSs-1 increased TNF-α and IL-6 levels and decreased IL-10. Meanwhile, PSDSs-2 showed the opposite effect, indicating the difference in pro- and anti-inflammatory activities of PSDSs with different molecular weights. The immunosuppressive model of cyclophosphamide proved that PSDSs have immune-promoting function, with PSDSs-1 exhibiting a better effect than PSDSs-2. In vitro and in vivo experiments illustrated the complexity of PSDS immunomodulation. Further research on the functions of PSDs with different molecular weights is needed to lay a foundation for their classification and application

TNF Induction of NF-κB RelB Enhances RANKL-Induced Osteoclastogenesis by Promoting Inflammatory Macrophage Differentiation but also Limits It through Suppression of NFATc1 Expression

<div><p>TNF induces bone loss in common bone diseases by promoting osteoclast formation directly and indirectly, but it also limits osteoclast formation by inducing expression of NF-κB p100. Osteoclast precursors (OCPs) are derived from M1 (inflammatory) and M2 (resident) macrophages. However, it is not known if TNF stimulates or limits osteoclast formation through regulation of M1 or M2 differentiation or if RelB, a partner of p100, is involved. To investigate these questions, we treated bone marrow cells (BMCs) with M-CSF alone or in combination with TNF to enrich for OCPs, which we called M-OCPs and T-OCPs, respectively. We found that TNF switched CD11b⁺F4/80⁺ M-OCPs from Ly6C^-Gr1^- M2 to Ly6C⁺Gr1^-CD11c⁺ and Ly6C^-Gr1^-CD11c⁺ M1 cells. RANKL induced osteoclast formation from both Ly6C⁺Gr1^- and Ly6C^-Gr1^- T-OCPs, but only from Ly6C⁺Gr1^- M-OCPs, which formed significantly fewer osteoclasts than T-OCPs. Importantly, Ly6C⁺Gr1^- cells from both M- and T-OCPs have increased expression of the M1 marker genes, iNOS, TNF, IL-1β and TGFβ1, compared to Ly6C^-Gr1^- cells, and Ly6C^-Gr1^- cells from T-OCPs also have increased expression of iNOS and TGFβ1 compared to cells from M-OCPs. Both RANKL and TNF increased RelB mRNA expression. TNF significantly increased RelB protein levels, but RANKL did not because it also induced RelB proteasomal degradation. TNF inhibited RANKL-induced NFATc1 mRNA expression and osteoclast formation from M-OCPs, but not from T-OCPs, and it did not induce Ly6C⁺Gr1^-CD11c⁺ or Ly6C^-Gr1^-CD11c⁺ M1 macrophages from RelB-/- BMCs. Furthermore, overexpression of RelB in M-OCPs reduced RANKL-induced osteoclast formation and NFATc1 mRNA expression, but it increased TNF-induced OC formation without affecting NFATc1 levels. Thus, TNF induction of RelB directly mediates terminal osteoclast differentiation independent of NFATc1 and limits RANKL-induced osteoclastogenesis by inhibiting NFATc1 activation. However, the dominant role of TNF is to expand the OCP pool by switching the differentiation of M-CSF-induced M2 to M1 macrophages with enhanced osteoclast forming potential. Strategies to degrade RelB could prevent TNF-induced M2/M1 switching and reduce osteoclast formation.</p></div

TNF-induced macrophages have higher OC forming potential than M-CSF-induced macrophages.

<p>(A) M-, T-, and R-OCPs cultured from BMCs from a 4-month-old C57Bl6 mouse were stained with the fluorescent-labeled antibodies as in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0135728#pone.0135728.g001" target="_blank">Fig 1</a>. Ly6C⁺Gr1^- and Ly6C^-Gr1^- populations from CD11b^<b>+</b>F4/80^<b>+</b> cells were sorted by flow cytometry. (B) The sorted cell populations were seeded in 96-well plates (4x10⁴ cells/well) and treated with RANKL or TNF in the presence of M-CSF for 2 additional days to generate mature OCs, which were stained for TRAP activity. (C) Quantitation of numbers of OCs formed from each sorted population in (B), 4 wells per group, *p < 0.05, **p < 0.01. The experiment was repeated twice with similar results. M = M-CSF, P = PBS, R = RANKL, T = TNF, R+T = RANKL+TNF.</p

Transcription Factors Responding to Pb Stress in Maize

Pb can damage the physiological function of human organs by entering the human body via food-chain enrichment. Revealing the mechanisms of maize tolerance to Pb is critical for preventing this. In this study, a Pb-tolerant maize inbred line, 178, was used to analyse transcription factors (TFs) expressed under Pb stress based on RNA sequencing data. A total of 464 genes expressed in control check (CK) or Pb treatment samples were annotated as TFs. Among them, 262 differentially expressed transcription factors (DETs) were identified that responded to Pb treatment. Furthermore, the DETs were classified into 4 classes according to their expression patterns, and 17, 12 and 2 DETs were significantly annotated to plant hormone signal transduction, basal transcription factors and base excision repair, respectively. Seventeen DETs were found to participate in the plant hormone signal transduction pathway, where basic leucine zippers (bZIPs) were the most significantly enriched TFs, with 12 members involved. We further obtained 5 Arabidopsis transfer DNA (T-DNA) mutants for 6 of the maize bZIPs, among which the mutants atbzip20 and atbzip47, representing ZmbZIP54 and ZmbZIP107, showed obviously inhibited growth of roots and above-ground parts, compared with wild type. Five highly Pb-tolerant and 5 highly Pb-sensitive in maize lines were subjected to DNA polymorphism and expression level analysis of ZmbZIP54 and ZmbZIP107. The results suggested that differences in bZIPs expression partially accounted for the differences in Pb-tolerance among the maize lines. Our results contribute to the understanding of the molecular regulation mechanisms of TFs in maize under Pb stress

Over-expression of RelB inhibits RANKL-, but enhances TNF-induced OC differentiation.

<p>(A) BMCs from 3-month-old C57Bl6 mice were cultured with M-CSF for 2 days followed by treatment of ¼ volume of pMX-GFP or pMX-GFP-RelB retroviral supernatant in the presence of 2 ng/ml of polybrene for 3 days. GFP⁺F4/80⁺ cells were analyzed by flow cytometry (left panel) and RelB protein levels in GFP or RelB expressing cells that had been treated with P, R or T for 8 hr were tested by Western blot (right panel). (B) M-OCPs were infected with pMX-GFP or pMX-GFP-RelB retrovirus as above. After 24 hr of infection, the cells were treated with RANKL or TNF for 3 days or 4 days in the presence of M-CSF when mature OCs were observed under inverted microscopy. TRAP staining was performed to evaluate OC numbers and area, 4 wells per group, *p< 0.05, **p< 0.01. (C) M-OCPs were infected with GFP or GFP-RelB retrovirus and cultured with P, R or T for 3 or 4 days as above (B). Total RNA was extracted from these cells using Trizol reagent, and mRNA expression of NFATc1 normalized to β-actin was tested by real-time PCR. *p< 0.05, **p< 0.01 vs. GFP. The in vitro experiment was repeated twice with similar results.</p

TNF increases expression of RelB mRNA and prevents RelB protein degradation.

<p>(A) M-, R-, and T- OCPs generated as in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0135728#pone.0135728.g001" target="_blank">Fig 1B</a> were treated with PBS (P), R, T or R+T for 8 hr or for 48 hr by which time mature OCs had formed. Cell lysates were subjected to Western blot analysis of RelB and β-actin. (B) M-OCPs were treated with P, R, T or R+T for 4 and 24 hr (left panel), or M-, R- and T-OCPs were treated with P, R, T or R+T for 48 hr by which time mature OCs had formed (right panel). Total RNA was extracted to test mRNA expression of NFATc1 normalized to β-actin. **p < 0.01 vs. the respective PBS-treated cells. (C) M-OCPs were serum-starved for 2 hr followed by treatment of P, R or T in the presence of 10 μM MG-132 for 3 hr. Protein levels of RelB and β-actin were tested by Western blot. The data are the band levels measured densitometrically, normalized to β-actin.</p

(+)-Usnic Acid Inhibits Migration of c-KIT Positive Cells in Human Colorectal Cancer

Inhibition of tumor cell migration is a treatment strategy for patients with colorectal cancer (CRC). SCF-dependent activation of c-KIT is responsible for migration of c-KIT positive [c-KIT(+)] cells of CRC. Drug resistance to Imatinib Mesylate (c-KIT inhibitor) has emerged. Inhibition of mTOR can induce autophagic degradation of c-KIT. (+)-usnic acid [(+)-UA], isolated from lichens, has two major functions including induction of proton shuttle and targeting inhibition of mTOR. To reduce hepatotoxicity, the treatment concentration of (+)-UA should be lower than 10 μM. HCT116 cells and LS174 cells were employed to investigate the inhibiting effect of (+)-UA (<10 μM) on SCF-mediated migration of c-KIT(+) CRC cells. HCT116 cells were employed to investigate the molecular mechanisms. The results indicated that firstly, 8 μM (+)-UA decreased ATP content via uncoupling; secondly, 8 μM (+)-UA induced mTOR inhibition, thereby mediated activation suppression of PKC-A, and induced the autophagy of the completed autophagic flux that resulted in the autophagic degradation and transcriptional inhibition of c-KIT and the increase in LDH release; ultimately, 8 μM (+)-UA inhibited SCF-mediated migration of CRC c-KIT(+) cells. Taken together, 8 μM could be determined as the effective concentration for (+)-UA to inhibit SCF-mediated migration of CRC c-KIT(+) cells