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Primary porcine tracheal epithelial cells (PTECs) are an appropriate model for studying the molecular mechanism of various porcine respiratory diseases, including swine-origin mycoplasmas, which are isolated from respiratory tract of pigs and mainly found on the mucosal surface surrounding swine trachea. However, the short proliferation ability of primary PTECs greatly limits their lifespan. In this study, primary PTECs were carefully isolated and cultured, and immortal PTECs were constructed by transfecting primary PTECs with the recombinant constructed plasmid pEGFP-hTERT containing human telomerase reverse transcriptase (hTERT). Immortal PTECs (hTERT-PTECs) maintained both the morphological and functional characteristics of primary PTECs, as indicated by the expression of cytokeratin 18, cell-cycle analysis, proliferation assay, Western blotting, telomerase activity assay, karyotype analysis and quantitative RT-PCR. Compared to primary PTECs, hTERT-PTECs had an extended replicative lifespan, higher telomerase activity, and enhanced proliferative activity. In addition, this cell line resulted in a lack of transformed and grown tumors in nude mice, suggesting that it could be safely applied in further studies. Moreover, hTERT-PTECs were vulnerable to all swine-origin mycoplasmas through quantitative analysis as indicated by 50% color changing unit (CCU50) calculation, and no significant differences of adhesion ability between primary and immortal PTECs were observed. For the representative swine mycoplasma Mycoplasma hyopneumoniae (Mhp), except for DNA copies quantitative real-time PCR assay, indirect immunofluorescence assay and Western blotting analysis also depicted that hTERT-PTECs was able to adhere to different Mhp strains of different virulence. In summary, like primary PTECs, hTERT-PTECs could be widely used as an adhesion cell model for swine-origin mycoplasmas and in infection studies of various porcine respiratory pathogens.

Direct conversion of biomass to functional materials is an ideal solution to relieve challenges in environmental and energy sustainability. We herein demonstrate a molten salt thermoelectrolysis of rice husks (RHs) mainly consisting of organic mass and biosilica to achieve high-efficiency and upgraded utilization of both Si and C in RHs. By coupling pyrolysis of organic mass with electrochemical reduction of silica in molten salts, the thermoelectrolysis of RHs in molten CaCl2–NaCl at 800 °C refines the RHs and acid-leached RHs to SiC nanowire/C (SiC-NW/C) and Si nanoparticle/C (Si-NP/C), respectively. The present study highlights the molten salt thermoelectrolysis for reclamation of biomass wastes in an affordable and controllable manner.

Monocyte migration into diseased tissues contributes to the pathogenesis of diseases. Intermediate-conductance Ca2+-activated K+ (KCa3.1) channels play an important role in cell migration. However, the role of KCa3.1 channels in mediating monocyte migration induced by palmitic acid (PA) is still unclear. Using cultured THP-1 cells and peripheral blood mononuclear cells from healthy subjects, we investigated the role and signaling mechanisms of KCa3.1 channels in mediating the migration induced by PA. Using methods of Western blotting analysis, RNA interference, cell migration assay and ELISA, we found that PA-treated monocytes exhibited increment of the protein levels of KCa3.1 channel and monocyte chemoattractant protein-1 (MCP-1), and the effects were reversed by co-incubation of PA with anti-TLR2/4 antibodies or by specific inhibitors of p38-MAPK, or NF-κB. In addition, PA increased monocyte migration, which was abolished by a specific KCa3.1 channel blocker, TRAM-34, or KCa3.1 small interfering RNA (siRNA). The expression and secretion of MCP-1 induced by PA was also similarly prevented by TRAM-34 and KCa3.1 siRNA. These results demonstrate for the first time that PA upregulates KCa3.1 channels through TLR2/4, p38-MAPK and NF-κB pathway to promote the expression of MCP-1, and then induce the trans-endothelial migration of monocytes.

Summary Reinvigoration of antitumor immunity remains an unmet challenge. Our retrospective analyses revealed that cancer patients who took antihistamines during immunotherapy treatment had significantly improved survival. We uncovered that histamine and histamine receptor H1 (HRH1) are frequently increased in the tumor microenvironment and induce T cell dysfunction. Mechanistically, HRH1-activated macrophages polarize toward an M2-like immunosuppressive phenotype with increased expression of the immune checkpoint VISTA, rendering T cells dysfunctional. HRH1 knockout or antihistamine treatment reverted macrophage immunosuppression, revitalized T cell cytotoxic function, and restored immunotherapy response. Allergy, via the histamine-HRH1 axis, facilitated tumor growth and induced immunotherapy resistance in mice and humans. Importantly, cancer patients with low plasma histamine levels had a more than tripled objective response rate to anti-PD-1 treatment compared with patients with high plasma histamine. Altogether, pre-existing allergy or high histamine levels in cancer patients can dampen immunotherapy responses and warrant prospectively exploring antihistamines as adjuvant agents for combinatorial immunotherapy.

How to revive anti-tumor immunity in cancer patients is an unmet challenge. In retrospective screening of common medications that may affect immunotherapy response in patients, we found that patients who took antihistamines during immunotherapy treatment had significantly improved survival. Further study uncovered that histamine and histamine receptor H1 (HRH1) are frequently increased in tumor microenvironment to induce T-cell dysfunction. Mechanistically, HRH1 activation in macrophages inhibits proinflammatory signaling, dysregulates fatty acid metabolism, and increases membrane expression of immune checkpoint V-domain Ig suppressor of T-cell activation (VISTA) in macrophage, which renders CD8+ T-cells dysfunctional. Knockout HRH1 or antihistamine treatment negated the immunosuppressive activity of the macrophages, revitalized T-cell cytotoxic function, and restored immunotherapy response. Fascinatingly, both animal and human data showed that allergy facilitated tumor growth and induced immunotherapy resistance via histamine/HRH1 axis, underscoring the tumor-prone activity of allergy. These findings demonstrate that pre-existing allergy in cancer patients can dampen response to immunotherapies and warrant perspective investigation of antihistamines as an adjuvant agent for combinatorial immunotherapy.

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) or agonistic antibodies targeting TRAIL-receptors (TRAIL-Rs) can selectively induce apoptosis in cancer cells. However, they have limited antitumor efficacy in clinical trials. We previously generated ten fully human monoclonal Abs to TRAIL-receptor type 1 (TR1-mAbs) using immunospot array assay on a chip (ISAAC technology). We found that the TR1-mAbs exhibited different effects on TRAIL-induced apoptosis (enhanced or blocked apoptosis). Here, we further demonstrated that some mAbs competed with TRAIL for binding to TRAIL-R1 expressed on tumor cells that blocked TRAIL-induced apoptosis (B-TR1-Ab), whereas others did not compete with TRAIL that enhanced TRAIL-induced apoptosis (E-TR1-Ab). Combination of E-TR1-Ab (TR1-419) with TRAIL leads to enhanced antitumor activity in various tumor cells in vitro. E-TR1-419 and TRAIL could cooperate to upregulate the mRNA expression and protein levels of TRAIL-R1 and to promote caspase-8 cleavage and increased JNK phosphorylation. Our results suggest that combining E-TR1 Ab with TRAIL could provide a new therapeutic strategy for tumor immunotherapies.

The jack-up rig is the most widely applied mobile platform in the shallow-water offshore drilling. Since several predetermined target wells are in the soil stratum of sand overlying clay, the punch-through accidents occur frequently during the jack-up rig spudcan penetration leading to disastrous consequences and sometimes even casualties. Although a few centrifugal model experiments and indoor large-scale experiments have been conducted to simulate the spudcan penetration, these experiments expose significant knowledge gaps that deviate from the actual field operations due to the limitations of a constant displacement speed plan instead of multi-stage preloading plan. In this study, seventeen (17) field experiments simulating spudcan installation in sand overlying clay in in-situ soil strata in the intertidal sea of the Bohai Bay are reported to address the knowledge gaps. A systematic study of the spudcan penetration is presented through key influencing factors such as soil strength ratio of upper sand to lower clay, geometric ratio of spudcan diameter to upper sand thickness, ballasting speed and holding period time. Results indicate that the ultimate (peak) soil bearing capacity increases almost quadratically with the increase in strength ratio of upper sand to lower clay; the final spudcan penetration resistance decreases as a power law relationship with the increase in geometric ratio of spudcan diameter to upper sand thickness; the instantaneous spudcan penetration velocity increases as a logarithmic function with the increase in ballasting speed and the pore water pressure decreases as an exponential function with the increase in holding period time. From these experiments, three novel schemes are proposed to prevent the punch-through. Lastly, this study provides a design basis and theoretical guidance for precise calculations of the ultimate bearing capacity, accurate analysis of punch-through risk and effective prevention of the punch-through accidents in the real jack-up rig.

Arsenic contamination has attracted worldwide concerns, owing to its toxicity and severe threat to human and environment. It is urgent to develop efficient adsorbents to remove arsenic pollutants. Within this paper, both pristine MIL-88A(Fe) and MIL-88A(Fe) decorated on cotton fibers were successfully fabricated using an eco-friendly method. The pristine MIL-88A(Fe) displayed outstanding adsorption performances towards four selected arsenic pollutants, in which the adsorption capacities toward As(III), As(V), ROX and ASA were 126.5, 164.0, 261.4 and 427.5 mg g−1, respectively. Additionally, MIL-88A(Fe) exhibited excellent removal efficiencies in a wide pH range and with the presence of different co-existing ions. It was proposed that the coordinative interactions of As–O–Fe between arsenic pollutants and MIL-88A(Fe) contributed to the superior adsorption performances. Furthermore, two MIL-88A(Fe)/cotton fibers composites were synthesized by both post synthesis (MC-1) and in-situ synthesis (MC-2), which demonstrated identically outstanding adsorption activities toward four selected arsenic pollutants. MC-1 and MC-2 enhanced the stability and reusability of MIL-88A(Fe), which was challenging issues of pristine MIL-88A(Fe) powder. Additionally, the fixed-bed column packed by MC-1 or MC-2 can continuously eliminate arsenic pollutants from the water flow. This work provided a new possibility of metal-organic frameworks to accomplish potentially large-scale application to purify the arsenic-contaminated water.

Background: Breast cancer patients at the same stage may show different clinical prognosis or different therapeutic effects of systemic therapy. Methods: The differentially expressed genes were identified of breast cancer from GSE42568. Through survival, ROC, random forest, GSVA and Cox regression model to identify genes that can be associated with survival time in breast cancer. The molecular mechanism was identified by enrichment and GSEA, methylation and SNV analysis. Then, the expression of a key gene was verified by TCGA dataset and RT-qPCR, Western blot, immunohistochemistry. Findings: We identified 784 genes related to the 5-year survival time of breast cancer. Through ROC curve and random forest, we screened 10 prognostic genes. Which were integrated into complex by GSVA, high expression of the complex significantly promoted the recurrence free survival of breast cancer patients. Enrichment analysis showed that key genes were related to immune and metabolic related functions. Importantly, we found methylation of MEX3A and TBC1D9, and mutations events. Finally, the expression of UGCG was verified by TCGA dataset and RT-qPCR, Western blot, immunohistochemistry in our own samples. Interpretation: The results indicate that the 10 genes may be potential biomarkers and therapeutic targets for long-term survival in breast cancer, especially UGCG. Funding Statement: This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors. Declaration of Interests: All authors declare no competing interests. Ethics Approval Statement: This study was approved by the human ethics review committee of Harbin Medical University. Ten cases of breast cancer were collected from the Cancer Institute of the Harbin Medical University Cancer Hospital with the informed consent of the patients.

ARID1A gene encodes BAF250a which is a member of the ARID family of DNA-binding proteins and a subunit of human SWI/SNF-related complexes. Low expression of ARID1A has been correlated with specific tumor cell lines or specific pathological types of cancer tissue. The purpose of this study was to investigate the expression of ARID1A in invasive ductal breast carcinomas and to evaluate its clinicopathological characteristics and prognostic value.ARID1A mRNA expression was evaluated by real-time reverse transcriptase-polymerase chain reaction (RT-PCR) in 40 pairs of fresh frozen breast cancer and normal breast samples. BAF250a expression was evaluated by immunohistochemistry in 112 paraffin-embedded surgical specimens of invasive breast cancers and 20 cases of matched normal breast tissues. We further analyzed the clinicopathological characteristics of ARID1A expression. Overall survival time was assessed by the Kaplan-Meier method and Cox regression model.ARID1A mRNA expression was lower in breast cancer tissue than in corresponding normal tissue (P0.001), and this decreased expression level was markedly associated with factors such as larger tumor size (P=0.038), higher stage (P=0.016), ER(-) (P=0.038), higher Ki-67 (P=0.025), P53 mutation (P=0.018) and ER(-)/PR(-)/Her-2(-) molecular subtype (P=0.044). With immunohistochemical staining, we showed that low BAF250a expression existed in 56% (63/112) of the breast cancers tissues. Low BAF250a expression was significantly associated with tumor stage (P=0.021), P53 (P=0.018), Ki-67 (P=0.031) and ER(-)/PR(-)/Her-2(-) molecular subtype (P=0.044). Low ARID1A expression was a predictor, not an independent, of overall survival.These data suggest that low ARID1A expression is frequent in breast cancers, and we need to investigate further the role of ARID1A and SWI/SNF complexes in breast tumorigenesis, especially in triple-negative breast cancer.

The aim was to describe the clinicopathological features and prognosis of young patients with breast cancer.We reviewed the records of 1478 consecutive patients aged ≤50 years with first diagnosis of invasive breast cancer referred to surgery from January 1999 to March 2005. A total of 174 patients were aged35 years (group I) and 1304 were aged 35-50 years (group II).Compared with patients of group II, patients of group I had a higher percentage of tumors classified as estrogen receptors (ER) negative, progesterone receptors (PR) negative, with a Ki-67 labeling index ≥20% of the cells. The 5-year survival of group I was 78.3% as compared with 84.2% for group II (P = 0.006).Compared with patients aged between 35 and 50 years, patients aged35 years have a greater chance of having an endocrine-unresponsive tumor and a significantly poor prognosis.