Neutrophil-Driven M2-Like Macrophages Are Critical for Skin Fibrosis in a Systemic Sclerosis Model.

Systemic sclerosis (SSc) is a challenging autoimmune disease characterized by progressive fibrosis affecting the skin and internal organs. Despite the known infiltration of macrophages and neutrophils, their precise contributions to SSc pathogenesis remain elusive. In this study, we elucidated that CD206hiMHCIIlo M2-like macrophages constitute the predominant pathogenic immune cell population in the fibrotic skin of a bleomycin-induced SSc mouse model. These cells emerged as pivotal contributors to the profibrotic response by orchestrating the production of TGF-ß1 through a MerTK signaling-dependent manner. Notably, we observed that neutrophil infiltration was a prerequisite for accumulation of M2-like macrophages. Strategies such as neutrophil depletion or inhibition of CXCR1/2 were proven effective in reducing M2-like macrophages, subsequently mitigating SSc progression. Detailed investigations revealed that in fibrotic skin, neutrophil-released neutrophil extracellular traps were responsible for the differentiation of M2-like macrophages. Our findings illuminate the significant involvement of the neutrophil-macrophage-fibrosis axis in SSc pathogenesis, offering critical information for the development of potential therapeutic strategies.

JUNO high purity nitrogen plant.

The Jiangmen Underground Neutrino Observatory (JUNO) is a 20 kt low level radioactivity liquid scintillator detector in a laboratory 650 m underground. An excellent energy resolution and a large volume offer exciting opportunities for addressing many important topics in neutrino physics. High purity nitrogen is an important factor to ensure the low background of the JUNO detector. High Purity Nitrogen (HPN) is used for detector purging, pipe cleaning, and scintillator purification, among other things in JUNO. According to JUNO's requirements, the radon concentration in HPN should be less than 10 µBq/m3. To meet this requirement, A high-purity nitrogen plant with 100 Nm3/h maximum rate was designed and constructed. Low-temperature adsorption technology is used to remove radioactive impurities in nitrogen. High purification efficiency was ensured by using an activated carbon column with high column height-to-diameter ratio. Electrostatic collection and low-temperature enrichment methods are combined to measure radon in nitrogen. After ten days of continuous operation at 50 Nm3/h flux rate, the plant can to reduce the radon concentration in nitrogen from 37.4±1.8µBq/m3 to less than 1.33 µBq/m3. After HPN with flow rate of 50 Nm3/h passing through low-background pipeline (About 1.3 km), the radon concentration of HPN is 5.6±0.6µBq/m3.

SOD2 promotes the immunosuppressive function of mesenchymal stem cells at the expense of adipocyte differentiation.

The potent immunomodulatory function of mesenchymal stem/stromal cells (MSCs) elicited by proinflammatory cytokines IFN-γ and TNF-α (IT) is critical to resolve inflammation and promote tissue repair. However, little is known about how the immunomodulatory capability of MSCs is related to their differentiation competency in the inflammatory microenvironment. In this study, we demonstrate that the adipocyte differentiation and immunomodulatory function of human adipose tissue-derived MSCs (MSC(AD)s) are mutually exclusive. Mitochondrial reactive oxygen species (mtROS), which promote adipocyte differentiation, were decreased in MSC(AD)s due to IT-induced upregulation of superoxide dismutase 2 (SOD2). Furthermore, knockdown of SOD2 led to enhanced adipogenic differentiation but reduced immunosuppression capability of MSC(AD)s. Interestingly, the adipogenic differentiation was associated with increased mitochondrial biogenesis and upregulation of peroxisome proliferator-activated receptor gamma coactivator-1 alpha (PPARGC1A/PGC-1α) expression. IT inhibited PGC-1α expression and decreased mitochondrial mass but promoted glycolysis in an SOD2-dependent manner. MSC(AD)s lacking SOD2 were compromised in their therapeutic efficacy in DSS-induced colitis in mice. Taken together, these findings indicate that the adipogenic differentiation and immunomodulation of MSC(AD)s may compete for resources in fulfilling the respective biosynthetic needs. Blocking of adipogenic differentiation by mitochondrial antioxidant may represent a novel strategy to enhance the immunosuppressive activity of MSCs in the inflammatory microenvironment.

Mesenchymal stromal cells confer breast cancer doxorubicin resistance by producing hyaluronan.

Chemotherapy resistance represents a major cause of therapeutic failure and mortality in cancer patients. Mesenchymal stromal cells (MSCs), an integral component of tumor microenvironment, are known to promote drug resistance. However, the detailed mechanisms remain to be elucidated. Here, we found that MSCs confer breast cancer resistance to doxorubicin by diminishing its intratumoral accumulation. Hyaluronan (HA), a major extracellular matrix (ECM) product of MSCs, was found to mediate the chemoresistant effect. The chemoresistant effect of MSCs was abrogated when hyaluronic acid synthase 2 (HAS2) was depleted or inhibited. Exogenous HA also protected tumor grafts from doxorubicin. Molecular dynamics simulation analysis indicates that HA can bind with doxorubicin, mainly via hydrophobic and hydrogen bonds, and thus reduce its entry into breast cancer cells. This mechanism is distinct from the reported chemoresistant effect of HA via its receptor on cell surface. High HA serum levels were also found to be positively associated with chemoresistance in breast cancer patients. Our findings indicate that the HA-doxorubicin binding dynamics can confer cancer cells chemoresistance. Reducing HA may enhance chemotherapy efficacy.

Nitric oxide-dependent immunosuppressive function of thymus-derived mesenchymal stromal/stem cells.

BACKGROUND: The thymus is required for T cell development and the formation of the adaptive immunity. Stromal cells, which include thymic epithelial cells (TECs) and mesenchymal stromal cells (MSCs), are essential for thymic function. However, the immunomodulatory function of thymus-derived MSCs (T-MSCs) has not been fully explored. METHODS: MSCs were isolated from mouse thymus and their general characteristics including surface markers and multi-differentiation potential were characterized. The immunomodulatory function of T-MSCs stimulated by IFN-γ and TNF-α was evaluated in vitro and in vivo. Furthermore, the spatial distribution of MSCs in the thymus was interrogated by using tdTomato-flox mice corssed to various MSC lineage Cre recombinase lines. RESULTS: A subset of T-MSCs express Nestin, and are mainly distributed in the thymic medulla region and cortical-medulla junction, but not in the capsule. The Nestin-positive T-MSCs exhibit typical immunophenotypic characteristics and differentiation potential. Additionally, when stimulated with IFN-γ and TNF-α, they can inhibit activated T lymphocytes as efficiently as BM-MSCs, and this function is dependent on the production of nitric oxide (NO). Additionally, the T-MSCs exhibit a remarkable therapeutic efficacy in acute liver injury and inflammatory bowel disease (IBD). CONCLUSIONS: Nestin-positive MSCs are mainly distributed in medulla and cortical-medulla junction in thymus and possess immunosuppressive ability upon stimulation by inflammatory cytokines. The findings have implications in understanding the physiological function of MSCs in thymus.

Particulate Standard Establishment for Absolute Quantification of Nanoparticles by LA-ICP-MS.

The development of nanotechnology has transformed many cutting-edge studies related to single-molecule analysis into nanoparticle (NP) detection with a single-NP sensitivity and ultrahigh resolution. While laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) has been successful in quantifying and tracking NPs, its quantitative calibration remains a major challenge due to the lack of suitable standards and the uncertain matrix effects. Herein, we frame a new approach to prepare quantitative standards via precise synthesis of NPs, nanoscale characterization, on-demand NP distribution, and deep learning-assisted NP counting. Gold NP standards were prepared to cover the mass range from sub-femtogram to picogram levels with sufficient accuracy and precision, thus establishing an unambiguous relationship between the sampled NP number in each ablation and the corresponding mass spectral signal. Our strategy facilitated for the first time the study of the factors affecting particulate sample capture and signal transductions in LA-ICP-MS analysis and culminated in the development of an LA-ICP-MS-based method for absolute NP quantification with single-NP sensitivity and single-cell quantification capability. The achievements would herald the emergence of new frontiers cut across a spectrum of toxicological and diagnostic issues related to NP quantification.

A Monte Carlo method for Rayleigh scattering in liquid detectors.

A new Monte Carlo method has been implemented to describe the angular and polarization distributions of anisotropic liquids, such as water and linear alkylbenzene (LAB), by considering orientational fluctuations of polarizability tensors. The scattered light of anisotropic liquids is depolarized with an angular distribution of 1 + (1 - ρv)/(1 + 3ρv) cos2 θ, which is modified by the depolarization ratio ρv. A standalone experiment has validated the simulation results of LAB. The new method can provide more accurate knowledge on light propagation in large liquid detectors, which is beneficial to the development of reconstruction for detectors.

Mesenchymal stem/stromal cells primed by inflammatory cytokines alleviate psoriasis-like inflammation via the TSG-6-neutrophil axis.

Psoriasis is currently an incurable skin disorder mainly driven by a chronic inflammatory response. We found that subcutaneous application of umbilical cord- derived mesenchymal stem/stromal cells (MSCs) primed by IFN-γ and TNF-α, referred to as MSCs-IT, exhibited remarkable therapeutic efficacy on imiquimod (IMQ)-induced psoriasis-like inflammation in mice. Neutrophil infiltration, a hallmark of psoriasis, was significantly reduced after treatment with MSCs-IT. We further demonstrated that the effects of MSCs-IT were mediated by tumor necrosis factor (TNF) stimulating gene-6 (TSG-6), which was greatly upregulated in MSCs upon IFN-γ and TNF-α stimulation. MSCs transduced with TSG-6 siRNA lost their therapeutic efficacy while recombinant TSG-6 applied alone could also reduce neutrophil infiltration and alleviate the psoriatic lesions. Furthermore, we demonstrated that TSG-6 could inhibit neutrophil recruitment by decreasing the expression of CXCL1, which may be related to the reduced level of STAT1 phosphorylation in the keratinocytes. Thus, blocking neutrophil recruitment by MSCs-IT or TSG-6 has potential for therapeutic application in human psoriasis.

Mesenchymal stem cells alleviate systemic sclerosis by inhibiting the recruitment of pathogenic macrophages.

Systemic sclerosis (SSc) is a recalcitrant autoimmune disease for which there is no cure. Mesenchymal stem cell (MSC)-based treatment has emerged as a promising therapeutic option for several autoimmune diseases. Previously, we found that the immunoregulatory potential of MSCs can be greatly enhanced by IFN-γ and TNF-α. Here, we found that IFN-γ- and TNF-α-pretreated MSCs significantly alleviated skin fibrosis in a bleomycin (BLM)-induced SSc model. Macrophages were found to be the predominant profibrotic immune cell population in the pathogenesis of SSc. The accumulation of macrophages was significantly decreased by MSC treatment. Importantly, MSCs primarily reduced the population of maturing macrophages with high CCR2 expression by inhibiting the generation of CCL2 from fibroblasts and macrophages. This finding may help to improve MSC-based clinical treatments for SSc patients.

TNFα and IFNγ rapidly activate PI3K-AKT signaling to drive glycolysis that confers mesenchymal stem cells enhanced anti-inflammatory property.

BACKGROUND: Mesenchymal stem/stromal cells (MSCs) acquire immunosuppressive capacity only in an inflammatory microenvironment. This can be recapitulated in vitro by treating MSCs with inflammatory cytokines TNFα and IFNγ, which induce indoleamine 2,3-dioxygenase (IDO) and TNF-stimulated gene-6 (TSG-6). However, the signaling pathways downstream of the cytokines remain to be elucidated. METHODS: Inflammatory bowel disease (IBD) mouse model was established by subjecting mice to dextran sulfate sodium (DSS) in drinking water for 7 days. Human UC-MSCs were pretreated with TNF-α and IFN-γ for 24 h and were then infused intravenously at day 2 of DSS administration. Colon tissues were collected for length measurement and histopathological examination. The serum level of IL-6 in mice was measured by enzyme-linked immunosorbent assay. Real-time PCR and Western blot were used to examine the mRNA level and protein expression. MSCs overexpressing constitutive active AKT or dominant negative AKT were generated and were analyzed. The glycolysis level of the MSCs was measured using Extracellular Flux Analyzer. 2-NBDG was used to monitor the uptake of glucose by MSCs. RESULTS: TNFα and IFNγ treatment led to rapid consumption of glucose and metabolic skewing toward glycolysis in MSCs, which was required for the therapeutic efficacy of MSCs on IBD. Blockade of glycolysis in MSCs inhibited the expression of immunomodulatory molecules, IDO and TSG-6, as well as the therapeutic effect on IBD. Moreover, PI3K-AKT signaling axis was rapidly activated and was required for the skewing toward glycolysis induced by TNFα and IFNγ. MSCs expressing dominant negative AKT were compromised in their therapeutic efficacy on IBD. CONCLUSION: The glycolysis-dependent anti-inflammatory property of MSCs conferred by inflammatory cytokines is mediated by PI3K-AKT signaling pathway.

Measurements of Rayleigh ratios in linear alkylbenzene.

This paper describes a new experimental setup designed for the direct measurement of the Rayleigh ratio and Rayleigh scattering length for linear alkylbenzene, a solvent commonly used in liquid scintillator detectors for neutrino experiments. Using the new approach, the perpendicularly polarized Rayleigh ratio was determined to be (4.52 ± 0.28) × 10-6 m-1 sr-1 at 405 nm and (3.82 ± 0.24) × 10-6 m-1 sr-1 at 432 nm, and the corresponding Rayleigh scattering length was LRay = 22.9 ± 0.3(stat.) ± 1.7(sys.) m at 405 nm and LRay = 27.0 ± 0.9(stat.) ± 1.8(sys.) m at 432 nm. These results are consistent with both previous results determined using other experimental strategies and theoretical predictions.

Impact of Albumin Pre-Coating on Gold Nanoparticles Uptake at Single-Cell Level.

Nanoparticles (NPs) suspension is thermodynamically unstable, agglomeration and sedimentation may occur after introducing NPs into a physiological solution, which in turn affects their recognition and uptake by cells. In this work, rod-like gold NPs (AuNRs) with uniform morphology and size were synthesized to study the impact of bovine serum albumin (BSA) pre-coating on the cellular uptake of AuNRs. A comparison study using horizontal and vertical cell culture configurations was performed to reveal the effect of NPs sedimentation on AuNRs uptake at the single-cell level. Our results demonstrate that the well-dispersed AuNRs-BSA complexes were more stable in culture medium than the pristine AuNRs, and therefore were less taken up by cells. The settled AuNRs agglomerates, although only a small fraction of the total AuNRs, weighed heavily in determining the average AuNRs uptake at the population level. These findings highlight the necessity of applying single-cell quantification analysis in the study of the mechanisms underlying the cellular uptake of NPs.

Two new metal-chalcogenide-cluster-based frameworks with single metal ions of Zn2+(/Sb3+) serving as inter-cluster linkers.

Two new metal-chalcogenide-cluster-based frameworks, in which P1-ZnSnS clusters are linked to each other by both corner-shared S2- ions and single metal ions of Zn2+ (or Sb3+) to form one new 3D (3,4)-connected network (MCCF-22) and one 2D-layered framework (MCCF-23), respectively, are reported. Notably, MCCF-22 exhibits good performance toward photodegradation of methylene blue compared with its analogue framework with only S2- ions as the linker.

Stable 3D neutral gallium thioantimonate frameworks decorated with transition metal complexes for a tunable photocatalytic hydrogen evolution.

Incorporating transition metal (TM) complexes into cluster-based chalcogenide frameworks is an effective synthetic strategy to induce structural diversity and control the optoelectronic properties, which may further improve their photocatalytic performance. However, limited studies have been conducted on frameworks constructed by TM complexes covalently bonded with supertetrahedral Tn clusters, let alone on their properties, especially photocatalytic H2 activity. Herein, three new isostructural three-dimensional (3D) neutral inorganic-organic open frameworks of gallium thioantimonate comprised of thiogallate-based supertetrahedral T3 clusters that are covalently bonded with TM complexes ([TM(TEPA)]2+, TM = Mn/Ni/Fe, TEPA = tetraethylenepentamine) at the edges and are linked by single Sb3+ ions at the corner, namely, NCF-3-Mn/Ni/Fe have been solvothermally synthesized and structurally characterized, and display good thermal and chemical stability. Benefiting from an adjustable TM centre, the title compounds possess tunable photocatalytic H2 evolution activity, among which NCF-3-Mn exhibits the highest photocatalytic activity probably due to its favourable band structure and enhanced carrier separation efficiency.

A novel copper-rich open-framework chalcogenide with chiral topology constructed from distinctive bimetallic [Cu5SnSe10] clusters.

Reported here is the first chiral copper-rich open-framework chalcogenide with a quartz (qtz) topology built on distinctive [Cu5SnSe10] clusters connected by [SnSe4] bridging units. Through in situ sulfur doping, sulfurized compounds could be obtained that exhibit improved photocatalytic performance. This work expands the family of COCs with new building blocks and topologies and demonstrates the significance of chalcogen doping in COCs.

Nanoparticles Determination by Laser Ablation Inductively Coupled Plasma Mass Spectrometry.

Quantitatively studying the biodistribution and transformation of nanomaterials is of great importance for nanotoxicological evaluation. Recently, laser ablation inductively coupled plasma mass spectrometry has been employed to distinguish nanoparticles (NPs) with their dissolved ions in biological samples. The principle of the proposal is based on a hypothesis that the intact NPs sampled by laser ablation will generate discrete sharp pulses of signals in ICP-MS measurement, being totally different from the continuous, relatively lower signals generated by ions. However, it is still a controversy whether NPs could maintain their intactness during the laser ablation. This work found a way to exactly determine the number of NPs sampled for each LA-ICP-MS measurement. It made possible to reveal the signal profile of a single NP in LA-ICP-MS analysis. The results suggest that AuNR, AgNP and TIO2 NP were broken into much smaller secondary NPs during the laser ablation, therefore generating continuous signals in the analyzer. There was a certain probability that the fragmentation of large-sized NP or multiple NPs by laser ablation was not sufficient, leaving some NPs unbroken or some secondary NPs with relatively large sizes to generate discrete pulses of signals in the analyzer. When the intactness of NPs during laser ablation cannot be assured, it is impossible to determine the attribution of mass spectrum signals. These findings compromise the reliability of distinguishing NPs from their dissolved ions by LA-ICP-MS.

Radiolabeling of Nanomaterials: Advantages and Challenges.

Quantifying the distribution of nanomaterials in complex samples is of great significance to the toxicological research of nanomaterials as well as their clinical applications. Radiotracer technology is a powerful tool for biological and environmental tracing of nanomaterials because it has the advantages of high sensitivity and high reliability, and can be matched with some spatially resolved technologies for non-invasive, real-time detection. However, the radiolabeling operation of nanomaterials is relatively complicated, and fundamental studies on how to optimize the experimental procedures for the best radiolabeling of nanomaterials are still needed. This minireview looks back into the methods of radiolabeling of nanomaterials in previous work, and highlights the superiority of the "last-step" labeling strategy. At the same time, the problems existing in the stability test of radiolabeling and the suggestions for further improvement are also addressed.

Antimony-Assisted Assembly of Basic Supertetrahedral Clusters into Heterometallic Chalcogenide Supraclusters.

Reported here are two new heterometallic chalcogenide supraclusters ([Ga56Sb16S136] and [In36Sb6S75]), which present new self-assembly modes of basic supertetraheral clusters with the assistance of antimony ions. This work demonstrates the crucial role of the added metal ions with nontetrahedral coordination geometry in developing supertetrahedral-cluster-based supraclusters with a new level of complexity.

Three new metal chalcogenide open frameworks built through co-assembly and/or hybrid assembly from supertetrahedral T5-InOS and T3-InS nanoclusters.

Reported here are three new metal chalcogenide open frameworks built from supertetrahedral [In35S52O8] (denoted as T5-InOS or o-T5) and [In10S20] (denoted as T3-InS) nanoclusters of different sizes and compositions via co-assembly and/or hybrid assembly modes. Such a set of cluster-based superlattices with dia topological structures clearly exhibit quantum size effects and electronic coupling interaction of adjacent nanoclusters, which can effectively explain that the band gap of the T3-(o-T5) hybrid-assembled material lies in the middle of T3-T3 and (o-T5)-(o-T5) co-assembled materials.

Influence of Surface Charge on the Phytotoxicity, Transformation, and Translocation of CeO2 Nanoparticles in Cucumber Plants.

The physiochemical properties of nanoparticles (NPs), including surface charge, will affect their uptake, transformation, translocation, and final fate in the environment. In this study, we compared the phytoxoxicity and transport behaviors of nano CeO2 (nCeO2) functionalized with positively charged (Cs-nCeO2) and negatively charged (PAA-nCeO2) coatings. Cucumber seedlings were hydroponically exposed to 0-1000 mg/L of Cs-nCeO2 and PAA-nCeO2 for 14 days and the contents, distribution, translocation, and transformation of Ce in plants were analyzed using inductively coupled plasma mass spectrometry, micro X-ray fluorescence (µ-XRF), and X-ray absorption near-edge spectroscopy (XANES), respectively. Results showed that the seedling growth and Ce contents in plant tissues were functions of exposure concentrations and surface charge. Cs-nCeO2 was adsorbed strongly on a negatively charged root surface, which led to significantly higher Ce contents in the roots and lower translocation factors of Ce from the roots to shoots in Cs-nCeO2 group than in PAA-nCeO2 group. The results of µ-XRF showed that Ce elements were mainly accumulated at the root tips and lateral roots, as well as in the veins and at the edge of leaves. XANES results revealed that the proportion of Ce(III) was comparable in the plant tissues of the two groups. We speculated that Cs-nCeO2 and PAA-nCeO2 were partially dissolved under the effect of root exudates, releasing Ce3+ ions as a result. Then, the Ce3+ ions were transported upward in the form of Ce(III) complexes along the vascular bundles and eventually accumulated in the veins. The other portion of Cs-nCeO2 and PAA-nCeO2 entered the roots through the gap of a Casparian strip at root tips/lateral roots and was transported upward as intact NPs and finally accumulated at the edge of the blade. This study will greatly advance our information on how the properties of NPs influence their phytotoxicity, uptake, and subsequent trophic transfer in terrestrial food webs.