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Transition metal sulfides/transition metal phosphides (TMS/TMP) has shown great potential in cancer diagnosis and treatment due to its unique structural, optical, acoustic and magnetic properties. TMS/TMP can be formed from sulfur/phosphorus source and metal into binary compounds, or from the interaction of hydrogen sulfide (or hydrogen sulfuric acid) with metal oxides or hydroxides. It has a series of unique properties, such as high conductivity, metalloid properties, a variety of valence states and adjustable structure and so on. These advantages make it have great potential in biomedical applications, such as diagnostic imaging, disease therapy and drug/gene delivery. This review presents the latest research progress of TMS/TMP on tumor imaging, diagnosis and treatment. Here, we first illustrate the synthesis approaches and surface modification of TMS/TMP. Then, its emerging applications in tumor diagnosis and therapy are highlighted. Moreover, the challenges of TMS/TMP-mediated diagnosis/treat are provided and the prospective for this field are discussed.

Background: Chronic neurologic diseases are common sequelae of COVID. They severely impact the quality of life and increase the burden on healthcare systems. The long COVID neurological symptoms are due to the robust replication of SARS-CoV-2 in the nasal neuroepithelial cells, leading to neuroinvasion and inflammation of the central nerve system (CNS). Currently used medications and vaccines do not inhibit the robust SARS-CoV-2 replication in the nasal epithelial cells. EGCG-palmitate (EC16), a multifunctional compound, has the potential to become a novel intranasal-delivered drug for minimizing post-COVID neurologic symptoms. Method: EC16-containing formulations were developed and tested in vitro against human β coronavirus OC43 (CoV-OC43) using a TCID50 assay following three test protocols differing in exposure sequence. Results: EC16 formulations in normal saline, phosphate buffered saline, and cell culture medium were found to effectively inhibit human β-coronavirus infection (>99.99%) after a 30-min contact. A single 10-min application to cells after infection (i.e., without direct contact with the virus) resulted in >99% inhibition of viral replication. Conclusion: With its antiviral, antioxidant, anti-inflammatory, and neuroprotective properties, EC16 in nasal formulations could be further developed for clinical applications to COVID-19 patients for minimizing long COVID neurological symptoms.

Unilateral posterior crossbite, one of the most frequent malocclusions, is often associated with functional lateral shift of the mandible. Although the effects of functional lateral shift on the mandible and temporomandibular joint have been examined in various animal experiments, cranial and maxillary changes have received less attention.The aim of this study was to investigate the effects of functional lateral shift on the craniofacial complex in growing rats.Eighty 5-week-old male Sprague-Dawley rats were randomly divided into an experimental group (n = 40), which received an oblique guide appliance that shifted the mandible to the left during closure, and a control group (n = 40). The rats were scanned by cone-beam computed tomography at 3 days and 1, 2, 4 and 8 weeks. The dimensions of the mandibular bone, condyle, maxilla and cranium were measured.The mandibles of rats in the experimental group were smaller than those of the rats in the control group and were asymmetrical. The condyles of the rats in the experimental group were thinner than those of the control rats. The condylar length on the ipsilateral side was shorter and wider than that on the contralateral side from 4 to 8 weeks. No significant differences in cranial length or height were observed between the experimental and control groups. The height of the upper first molar and alveolar bone on the contralateral side was significantly smaller than that on the ipsilateral side and in the controls from 4 to 8 weeks.Functional shift in the mandible produces morphological asymmetries in the mandible and maxillary region and may cause bilateral condylar degenerative changes.

Background: The number of people with diabetes is increasing, and many patients have significantly impaired cognitive function. For patients with diabetic encephalopathy (DE), simply lowering blood sugar does not improve learning and memory. Studies have shown that β-asarone can significantly improve cognitive impairment in patients with DE, but the specific mechanism of action is unclear. Objective: This experiment hopes to use a variety of experimental methods to clarify the protective effect and mechanism of β-asarone on brain neurons during the development of DE disease. Methods: A high-sugar and high-fat diet and streptozotocin injection-induced DE rat model was used. β-asarone was administered for four weeks. The experiment used the Morris water maze test, biochemical index detection, and many methods to evaluate the neuroprotective effect of β-asarone on DE rats from various aspects and understand its mechanism. Results: β-asarone reduced neuronal cell damage and significantly improved the learning and memory ability of DE rats. In addition, β-asarone can reduce the oxidative stress response and amyloid-β accumulation in the brain of DE model rats and increase the content of brain-derived neurotrophic factor (BDNF) in the brain tissue, thereby reducing neuronal cell apoptosis and playing a protective role. Conclusion: β-asarone can reduce the accumulation of oxidative stress and amyloid-β in the brain, increase the content of BDNF, reduce the apoptosis of neuronal cells, and exert neuronal protection, thereby improving the learning and memory ability of DE model rats.

Objective. The purpose of this study was to explore the molecular mechanism of Danggui Buxue Decoction (DBD) intervening premature ovarian failure (POF). Methods. The active compounds-targets network, active compounds-POF-targets network, and protein-protein interaction (PPI) network were constructed by a network pharmacology approach: Gene Ontology (GO) function and Kyoto Encyclopedia of Gene and Genome (KEGG) pathway analysis by DAVID 6.8 database. The molecular docking method was used to verify the interaction between core components of DBD and targets. Then, High-Performance Liquid Chromatography (HPLC) analysis was used to determine whether the DBD contained two key components including quercetin and kaempferol. Finally, the estrous cycle, organ index, ELISA, and western blot were used to verify that mechanism of DBD improved POF induced by cyclophosphamide (CTX) in rats. Results. Based on the network database including TCMSP, Swiss Target Prediction, DisGeNET, DrugBank, OMIM, and Malacard, we built the active compounds-targets network and active compounds-POF-targets network. We found that 2 core compounds (quercetin and kaempferol) and 5 critical targets (TP53, IL6, ESR1, AKT1, and AR) play an important role in the treatment of POF with DBD. The GO and KEGG enrichment analysis showed that the common targets involved a variety of signaling pathways, including the reactive oxygen species metabolic process, release of Cytochrome C from mitochondria and apoptotic signaling pathway, p53 signaling pathway, the PI3K-Akt signaling pathway, and the estrogen signaling pathway. The molecular docking showed that quercetin, kaempferol, and 5 critical targets had good results regarding the binding energy. Chromatography showed that DBD contained quercetin and kaempferol compounds, which was consistent with the database prediction results. Based on the above results, we found that the process of DBD interfering POF is closely related to the balance of ESR and AR in TP53-AKT signaling pathway and verified animal experiments. In animal experiments, we have shown that DBD and its active compounds can effectively improve estrus cycle of POF rats, inhibit serum levels of FSH and LH, protein expression levels of Cytochrome C, BAX, p53, and IL6, and promote ovary index, uterine index, serum levels of E2 and AMH, and protein expression levels of AKT1, ESR1, AR, and BCL2. Conclusions. DBD and its active components could treat POF by regulating the balance of ESR and AR in TP53-AKT signaling pathway.

The problem of approximate k -nearest neighbor search in high-dimensional space is a fundamental problem in many applications. We have observed that most existing approaches are unable to leverage the query results to improve the search performance. To address this limitation, we present a new index, called HCTree+, that aims to improve the query performance based on incoming queries and their results. First, we adopt a simple yet effective index to support efficient search. Second, incoming queries and their results are used to optimize the index dynamically for future queries. The experimental study shows that HCTree+ outperforms the state-of-the-art algorithms in terms of accuracy while achieving the desired efficiency.

Background Today, ultrasound is increasingly utilized in enzyme modification. Strongly dependent on the specific operational conditions, the modification effect brought by ultrasound can be activation and inactivation of enzymes. This work aims to study the ultrasound mechanisms under different conditions, to investigate the respective roles of free radical effect and mechanical effect in pectinase activation and inactivation, and to reveal the influence of pectinase concentration on the ultrasound-modification effect. Results When ultrasound was introduced to a liquid system, generation of free radicals was positively correlated with ultrasound intensity and treatment duration, but negatively correlated with temperature. Thiourea with a concentration of 4 mmol L-1 was selected as a free radical scavenger to effectively shield ultrasound free radicals. The highest enzyme activity of pectinase solutions at 0.1, 1.0, and 10.0 mg mL-1 was obtained at the same ultrasound intensity of 4.50 W mL-1 and time of 15 min, where the enzyme activity was increased by 68.24%, 20.98% and 18.83%, respectively. Furthermore, the addition of thiourea enhanced the enzyme activity at each tested ultrasound intensity and time, especially those exceeding the best conditions; it also eliminated the redshift phenomenon that was previously presented in the fluorescence spectra of pectinase samples. Conclusion Pectinase concentrations did not change the optimum ultrasound conditions for enzyme modification, but pectinase with a low concentration was more vulnerable to ultrasound treatment. During modification, ultrasound mechanical effects dominated in the pectinase activation, while free radical effects dominated in the inactivation process. © 2020 Society of Chemical Industry.

Two kinds of four-bladed perylene diimide (PDI) propellers with d8 metal and non-metal cores are efficiently synthesized. The Ni-PDI, Pd-PDI, and Pt-PDI propellers, equipped with d8 metal cores, have two absorption bands at 350–650 nm and 780–1200 nm with deep LUMO levels of −4.40 eV to −4.51 eV. The TTF-PDI, QU-PDI, and PH-PDI propellers with non-metal cores have only one absorption band at 350–650 nm with upshifted LUMO levels. Interestingly, the organic photovoltaic (OPV) results show that reducing the intramolecular charge traps between the blade and core subunits of the PDI propellers can effectively improve the power conversion efficiency (PCE). The device based on the QU-PDI acceptor exhibits a PCE that is up to more than 300 times higher (9.33%) than that of the d8 metal core PDI propellers (Pd-PDI, PCE = 0.03%), which is one of the best photovoltaic performances with an excellent fill factor (FF = 71.8%) exhibited by PDI-derivative acceptors. Conversely, the electro-catalytic H2 evolution activity of Pt-PDI (current destiny = 10.00 mA cm−2 at −0.377 V), which exhibited a record performance for PDI-based catalysts to date, is up to 1000 times greater than that of the non-metal core PDI propellers (QU-PDI, 0.01 mA cm−2 at −0.377 V). Our results indicate that both highly efficient OPV and electrochemical H2 evolution catalysts can be achieved via the rational functionalization of PDI propellers with non-metal and d8 metal cores.

Objective. The purpose of the present study was to explore the mechanism of Astragalus membranaceus in the treatment of sepsis. Methods. We searched the active components and targets of Astragalus membranaceus using the TCMSP and BATMAN databases. Then, the GeneCards, MalaCards, and OMIM databases were used to screen out relevant targets of sepsis. The common targets of the former two gene sets were uploaded to the STRING database to create an interaction network. DAVID was used to perform KEGG enrichment analysis of the core targets. Based on the results of KEGG and previous studies, key pathways for the development of sepsis were identified and experimentally validated. Result. We obtained 3,370 sepsis-related targets in databases and 59 active components in Astragalus membranaceus through data mining, corresponding to 1,130 targets. The intersection of the two types of targets led to a total of 318 common targets and 84 core targets were obtained after screening again. The KEGG and previous studies showed that these 84 core targets were involved in sepsis by regulating TNF, MAPK, and PI3K pathways. TNF, MAPK8, NF-κB, and IκBα are crucial in sepsis. Experimental validation demonstrated that some markers in sepsis model rats were improved after the intervention with Astragalus granules and their chemical components. Among them, IL-1β, IL-6, and TNF-α in rat serum were reduced. The mRNA and protein expression of TNF-α, IL-6, MMP9, MAPK8, and NF-κB were reduced in rat blood. However, the mRNA and protein expression of IκBα and PI3K were increased in rat blood. Conclusion. The AST could affect the TNF, PI3K, and MAPK pathway cascade responses centred on IκBα and NF-κB, attenuate the expression of IL-6 and MMP9, and interfere with the inflammatory response during sepsis.

Elevated intraocular pressure (IOP) is the only modifiable risk factor for primary open-angle glaucoma (POAG). Herein we sought to prioritize a set of previously identified IOP-associated genes using novel and previously published datasets. We identified several genes for future study, including several involved in cytoskeletal/extracellular matrix reorganization, cell adhesion, angiogenesis, and TGF-β signaling. Our differential correlation analysis of IOP-associated genes identified 295 pairs of 201 genes with differential correlation. Pathway analysis identified β-estradiol as the top upstream regulator of these genes with ESR1 mediating 25 interactions. Several genes (i.e., EFEMP1, FOXC1, and SPTBN1) regulated by β-estradiol/ESR1 were highly expressed in non-glaucomatous human trabecular meshwork (TM) or Schlemm’s canal (SC) cells and specifically expressed in TM/SC cell clusters defined by single-cell RNA-sequencing. We confirmed ESR1 gene and protein expression in human TM cells and TM/SC tissue with quantitative real-time PCR and immunofluorescence, respectively. 17β-estradiol was identified in bovine, porcine, and human aqueous humor (AH) using ELISA. In conclusion, we have identified estrogen receptor signaling as a key modulator of several IOP-associated genes. The expression of ESR1 and these IOP-associated genes in TM/SC tissue and the presence of 17β-estradiol in AH supports a role for estrogen signaling in IOP regulation.

White sponge nevus (WSN) is a rare autosomal dominant disease with a family history, often caused by mutations of the keratin 4 (K4) and keratin 13 (K13) genes in patients. It is characterized by frequently occurred white corrugated folds in the bilateral buccal mucosa with soft texture. On histopathological examination, hyperkeratosis of epithelial cells, edema, and vacuolar changes in the spinous cells are observed in the lesions, despite a normal layer of basal cells. WSN should be differentiated from other oral white spot diseases, mainly oral lichen planus, oral candidiasis, oral white edema, and Heck’s disease, to reduce misdiagnosis and unnecessary treatment. At present, there is no specific treatment method. The purpose of this study was to report the clinical data of four WSN patients of the same family with the K4 gene mutation. The occurrence of WSN in a pair of monozygotic twins with very similar clinical presentations was identified for the first time. The gene sequencing results showed that there was a heterozygous deletion (C. 438_440delCAA) in exon 1 of the K4 gene, resulting in an aspartic acid loss in both the proband and his father. Finally, the etiology, pathogenesis, pathological manifestations, clinical manifestations, diagnosis, differential diagnosis, and related treatment methods are discussed to provide a reference for clinical treatment of the disease.

Mandibular deviation affects the biomechanical environment of the temporomandibular joint (TMJ) and causes thinning of cartilage on the deviated side. We aimed to evaluate, using a rat model, the effect of mandibular functional deviation on the TMJ in relation to the functional roles of integrin β family members. The effects of experimental functional deviation on the TMJ of 6-week-old Sprague-Dawley female rats, randomly assigned to control (n = 42) and experimental groups (n = 42), were evaluated at 3 days and 1, 2, 4, and 8 weeks by histological staining, immunofluorescence, real-time quantitative polymerase chain reaction, and micro-computed tomography. The results showed that the experimental functional shift changed the shape of condyles, thinned the cartilage, and increased the proportion of the hypertrophic layer on the deviated sides of condyles. In addition, the extracellular matrix of the condyle cartilage exhibited degradation at 1 week and subchondral trabecular bone was lost at 4 and 8 weeks. Osteoarthritis (OA)-like changes occurred in the left and right condyles of rats in the experimental group and were aggravated over time. Integrin β family expression, especially integrin β

In order to reduce mid–far-infrared emissivity while enhancing the near-infrared absorption properties of polymer coating systems, bimetal coordination conjugated polymer coatings are fabricated via baking organic linkers (oligomer-containing nickel bis(dithiolene) moiety, 3,3′-diaminobenzidine, metal salts) in the presence of co-reagents (dimethylacetamide and acetic acid). By varying the reaction temperature, feed ratios and quantity of metals, the degree of cross-linking of the metallopolymer (MOP) coating could be precisely controlled with an excellent film-forming property. The structures were confirmed by FT-IR, XPS and ICP. The results indicate a metal (Cu, La, Sm and Lu) coordination effect with conjugated polymer main chains being formed together with nickel-bis(dithiolene) structures. Due to the high electron deficiency ability of the nickel bis(dithiolene) moiety, all MOP coatings show electron transport behaviors without hole transport properties, except for NiCu-PTBN and NiSm-PTBN. Doping of CuBr2 and Sm(OAc)3·6H2O with a metal coordination effect under carefully controlled conditions gives rise to NiCu-PTBN and NiSm-PTBN coatings with hole mobilities up to 2.9 × 10−4 cm2 V−1 s−1 and 2.5 × 10−4 cm2 V−1 s−1, respectively. Stealth performance experiments show that the absorption wavelength of the MOP coating is 750–1200 nm in the near-infrared range, which indicates its excellent near-infrared shield properties. The mid–far-infrared emissivity (2–22 μm) of the nickel/samarium-containing coating could decrease to 0.47 ± 0.03. Overall, these results raise the possibility that an MOP coating is a prospective candidate for infrared stealth applications.

Graft-versus-host disease (GVHD) is one of the most common and serious complications after allogeneic hematopoietic stem cell transplantation (allo-HSCT). About 45%∼83% of patients develop GVHD in the oral cavity. There has no medical records of oral submucous fibrosis (OSF) induced by GVHD after allo-HSCT, which should be brought to the attention of dentists.

The purpose of the present study was to explore the mechanism ofWe searched the active components and targets ofWe obtained 3,370 sepsis-related targets in databases and 59 active components inThe AST could affect the TNF, PI3K, and MAPK pathway cascade responses centred on I

With the shortage of fossil energy, wind energy has attracted people's attention increasingly. However, the intermittence nature of wind energy will lead to unstable power output, which may bring much challenge when wind power in integrated to grid. Thus, wind-hydro combination system which consists of wind farms and pumped storage power stations is established as one of the solutions to solve this problem. Due to high investment cost of energy storage system like pumped storage power stations, some improvements and optimizations of system capacity should be considered. In this paper, high-frequency and difficult-to-handle wind power are processed by the pumped storage power station and the remaining wind power are processed by conventional hydropower stations. Hilbert Huang transform(HHt) in this paper is used to decompose the wind power frequency. Then, 50 different capacities for two types of hydropower stations are calculated by genetic algorithm to improve the stability of integrated system output. The weight coefficient named OD(optimization design) is proposed to quantify the economy and stability of the combination system, which can understand the comparison of different capacity ratios in a clear way.

Multi-set query is a fundamental problem in computer systems and applications. Most traditional solutions for multi-set query are based on hash tables or bloom filters. However, when the sizes of multi-sets are large, these solutions cannot achieve small memory usage, fast query speed and high accuracy at the same time. In this work, we study the problem of using a learned oracle to improve the performance of traditional multi-set query processing empirically. The key idea is to train an oracle to predict which set contains a query item e as a classification problem. To ensure an exact query result, we combine the learned oracle with a standard bloom filter and an exact-match index to catch items that are not correctly identified by the oracle. When the oracle is both small and efficient, the whole query performance can be improved. In our framework, we treat the learned oracle as a complete black box, and is not dependent on its inner workings. Theoretical proofs and experimental results show that compared to the state-of-the-art, the error rate of our approach is 0% even with much less memory usage and a comparable speed.

Purpose Intraocular pressure (IOP), the primary risk factor for primary open-angle glaucoma, is determined by resistance to aqueous outflow through the trabecular meshwork (TM). IOP homeostasis relies on TM responses to mechanical stretch. To model the effects of elevated IOP on the TM, this study sought to identify coding and non-coding RNAs differentially expressed in response to mechanical stretch. Methods Monolayers of TM cells from non-glaucomatous donors (n = 5) were cultured in the presence or absence of 15% mechanical stretch, 1 cycle/second, for 24 hours using a computer-controlled Flexcell unit. We profiled mRNAs and lncRNAs with stranded total RNA sequencing and microRNA (miRNA) expression with NanoString-based miRNA assays. We used two-tailed paired t-tests for mRNAs and long non-coding RNAs (lncRNAs) and the Bioconductor limma package for miRNAs. Gene ontology and pathway analyses were performed with WebGestalt. miRNA-mRNA interactions were identified using Ingenuity Pathway Analysis Integrative miRNA Target Finder software. Validation of differential expression was conducted using droplet digital PCR. Results We identified 219 mRNAs, 42 miRNAs, and 387 lncRNAs with differential expression in TM cells upon cyclic mechanical stretch. Pathway analysis indicated significant enrichment of genes involved in steroid biosynthesis, glycerolipid metabolism, and extracellular matrix-receptor interaction. We also identified several miRNA master regulators (miR-125a-5p, miR-30a-5p, and miR-1275) that regulate several mechanoresponsive genes. Conclusions To our knowledge, this is the first demonstration of the differential expression of coding and non-coding RNAs in a single set of cells subjected to cyclic mechanical stretch. Our results validate previously identified, as well as novel, genes and pathways.

Cardiac mesenchymal stem cells (C-MSCs) are endogenous cardiac stromal cells that play a crucial role in maintaining normal cardiac function. Rab27b is a member of the small GTPase Rab family that controls membrane trafficking and the secretion of exosomes. However, its role in regulating energy metabolism in C-MSC is unclear. In this study, we analyzed mitochondrial oxidative phosphorylation by quantifying cellular oxygen consumption rate (OCR) and quantified the extracellular acidification rate (ECAR) in C-MSC with/without Rab27b knockdown. Knockdown of Rab27b increased glycolysis, but significantly reduced mitochondrial oxidative phosphorylation (OXPHOS) with loss of mitochondrial membrane potential in C-MSC. Furthermore, knockdown of Rab27b reduced H3k4me3 expression in C-MSC and selectively decreased the expression of the essential genes involved in β-oxidation, tricarboxylic acid cycle (TCA), and electron transport chain (ETC). Taken together, our findings highlight a novel role for Rab27b in maintaining fatty acid oxidation in C-MSCs.

Data dependencies have been widely used in error detection. However, errors might not be detected when the target data set is sparse and no conflicts occur. With a rapid increase in the number of data sources available for consumption, we consider how to apply both external data sources and matching dependencies (a general form of FD) to detect more potential errors in target data. However, accessing all the sources for error detection is impractical when the number of sources is large. In this demonstration, we present an efficient source selection algorithm that can select a proper subset of sources for error detection. A key challenge of this approach is how to estimate the gain of each source without accessing their datasets. To address the above problem, we develop a two-level signature mechanism for estimating the gain of each source. Empirical results on both real and synthetic data show high performance on both the effectiveness and efficiency of our algorithm.

Through the analysis of the engineering management training system, we know that the courses in the system can be divided into theoretical, practical and experimental courses. We aims to develop an examination management system by restructuring the evaluation process of the three types of courses. And the organizational structure, hardware architecture, and software architecture of the test management system are designed to be applied in the entire process of system development, operation and maintenance. Besides, under the influence of “the pneumonia caused by novel coronavirus, COVID-19” on education, we wish that the examination management system is more suitable for the needs of the macro environment, and the results can be used to the training and evaluation of other professionals.

Flexible and stable near-infrared (NIR) absorbing polymer films were prepared from the corresponding poly(aryl ether) precursors through subsequent ligand-exchange post-polymerization modification. The additional ligand triggers the ligand-exchange reaction during the post-polymerization modification process, which can significantly enhance the polymer yield, as well as the coordination ratio of the nickel-bis(dithiolene) segment in the polymer backbone by suppressing cross-linking side reactions. The presented ligand-exchange post-polymerization modification overcomes the limitations of the post-polymerization modification reaction, i.e. a high degree of cross-linking between the same reactive groups, and enhances the modification methods of analogous polymers. Upon incorporating the nickel-bis(dithiolene) moiety into the polymer backbone, the resulting polymers exhibit both strong near-infrared absorption at approximately 1200 nm with e > 104 mol−1 L cm−1 in an NMP solution and excellent free-standing film formation properties. The long-term aging-resistance test indicates that the incorporation of the nickel-bis(dithiolene) moiety into the polymer chain via chemical bonding is superior to physical blending for stabilizing the nickel-bis(dithiolene) moiety. Although the absorbance of the micromolecular NIR dyes Ni–O and Ni–N at the maximum wavelength gradually decreased up to 34% during the aging test (70 °C for 144 h), it should be noted that the NiO-P6 (10 mol% nickel-bis(dithiolene) segments) and NiN-P8 (34 mol% nickel-bis(dithiolene) segments) polymer films exhibited no decrease in the NIR absorption range. Finally, this work shows that the post-polymerization modification between the same reactive groups could also be an effective approach to install functional groups in a polymer backbone while avoiding a high degree of cross-linking side reactions, as well as offers an approach to design near-infrared absorbing polymeric materials with balanced near-infrared absorption and solution-processing properties.

Purpose Schlemm’s canal (SC) endothelial cells derived from donors with or without glaucoma showed different mechanical properties and gene expression. As an important contributor to the regulation of intraocular pressure (IOP) and pathogenesis of primary open-angle glaucoma (POAG), the heritable key epigenetic changes, methylation may play an important role in the physiologic function of SC cells. This study aims to identify differentially methylated CpG sites (DMSs) in primary cultures of human SC cells with or without glaucoma. Methods We examined the methylation pattern of seven strains of primary human cells (two glaucoma and five normal SC cell samples), which were isolated and characterized using established protocols. DNA methylation was profiled using Illumina Human Methylation 450 BeadChip. Raw data were extracted and exported using Illumina GenomeStudio software. After quantile normalization, DNA methylation data were analyzed using R package RnBeads in Bioconductor. DMSs were filtered with p ≤ 1E-5, methylation change ≥ 0.1, and false discovery rate ≤ 0.05. The closest genes and the location of each CpG site were annotated using R package FDb.InfiniumMethylation.hg19. Gene Ontology and pathway analysis was performed using WebGestalt. Selected DMSs were validated using the Zymo qMethyl kit. Results We used five non-glaucoma and two glaucomatous SC cell samples to profile genome-wide DNA methylation using Illumina Infinium Methylation BeadChips. Principle component analysis showed the separation between the glaucoma and control samples. After quality control and differential analysis, we identified 298 highly significant DMSs (p ≤ 1E-5). Among them, 221 DMSs were within 1 kb of a nearby gene. Gene Ontology analysis demonstrated significant enrichment in positive regulation of cell migration, negative regulation of endothelial cell proliferation, and stress fiber and actin filament bundles. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis showed enrichment in cell adhesion and gap junctions. Several glaucoma-related genes were identified, including TGFBR3, THBS1, PITX2, DAXX, TBX3, TNXB, ANGPT1, and PLEKHA7. We also examined differentially methylated regions (DMRs) near these CpG sites and identified significant DMRs in TBX3, TNXB1, DAXX, and PITX2. Conclusions This study represents the first genome-wide DNA methylation profiling in cultured human primary SC cells. The DMSs were enriched in the pathways related to outflow resistance. Several DMRs were validated in glaucoma-associated genes, further suggesting the role of DNA methylation in glaucoma development. This study could provide comprehensive understanding of DNA methylation in glaucoma and its effect on aqueous humor outflow.

Exosomes are critical mediators of intercellular crosstalk and regulator of cellular/tumor microenvironment. Exosomes have great prospects for clinical application as theranostic and prognostic probe. Nevertheless, the advancement of the exosomes research has been thwarted by limited knowledge elucidating the most efficient isolation method and theirin vivotrafficking. Here we have showed that combination of two size-based methods using 0.20 µm syringe filter and 100k centrifuge membrane filter followed by ultracentrifugation method yields a greater number of uniform exosomes. We also demonstrated the visual representation and quantification of differentialin vivodistribution of radioisotope131I-labelled exosomes from diverse cellular origins, e.g., tumor cells with or without treatments (HET0016 and GW2580), myeloid-derived suppressor cells and endothelial progenitor cells. We also determined that the distribution was dependent on the protein/cytokine contents of the exosomes. The appliedin vivoimaging modalities can be utilized to monitor disease progression, metastasis, and exosome-based targeted therapy.AbbreviationsbFGFbasic fibroblast growth factorCSF1Rcolony stimulating factor 1 receptorCTcomputed tomographyCTLA4cytotoxic T-lymphocyte-associated protein 4EGFepidermal growth factorEMTepithelial to mesenchymal transitionEVsextracellular vesiclesEPCsendothelial progenitor cellsFasLFas ligandG-CSFgranulocyte-colony stimulating factorGM-CSFgranulocyte-macrophage colony-stimulating factorHGFhepatocyte growth factorHSPheat shock proteinICAM-1intercellular adhesion molecule 1IFN-gammainterferon gammaIL – 1betainterleukin-1 betaIL – 1rainterleukin-1 receptor antagonistIL – 2interleukin-2IL – 4interleukin-4IL – 6interleukin-6IL – 7interleukin-7IL – 10interleukin-10IL – 12interleukin-12IL – 13interleukin-13IL – 17interleukin-17KCkeratinocyte-derived chemokineLIXlipopolysaccharide-induced CXC chemokineM-CSFmacrophage colony-stimulating factorMCP-1monocyte chemoattractant protein 1MDCmacrophage-derived chemokineMDSCsmyeloid derived suppressor cellsMFPmammary fat padMIP-1αmacrophage-inflammatory protein-1alphaMMP-2matrix metalloproteinase-2MRImagnetic resonance imagingNISsodium iodide symporterNTAnanoparticle tracking analysisPETpositron emission tomographyPF-4platelet factor 4RANTESregulated on activation, normal T cell expressed and secretedROIsregion of interestSDF-1αstromal cell-derived factor-1SEMstandard error of the meanSPECTsingle-photon emission computed tomographySCFstem cell factorTAMstumor-associated macrophagesTEMtransmission electron microscopyTIMP 2tissue inhibitors of metalloproteinases 2TLPCthin layer paper chromatographyTMEtumor microenvironmentTNF-αtumor necrosis factor-αTSLPthymic stromal lymphopoietinUCultracentrifugationVEGF-Avascular endothelial growth factor AVEGFR2vascular endothelial growth factor receptor 2.

Exosomes are critical mediators of intercellular crosstalk and are regulator of the cellular/tumor microenvironment. Exosomes have great prospects for clinical application as a theranostic and prognostic probe. Nevertheless, the advancement of exosomes research has been thwarted by our limited knowledge of the most efficient isolation method and their in vivo trafficking. Here we have shown that a combination of two size-based methods using a 0.20 μm syringe filter and 100k centrifuge membrane filter followed by ultracentrifugation yields a greater number of uniform exosomes. We also demonstrated the visual representation and quantification of the differential in vivo distribution of radioisotope (131)I-labeled exosomes from diverse cellular origins, e.g., tumor cells with or without treatments, myeloid-derived suppressor cells and endothelial progenitor cells. We also determined that the distribution was dependent on the exosomal protein/cytokine contents. The applied in vivo imaging modalities can be utilized to monitor disease progression, metastasis, and exosome-based targeted therapy.

This study reported a quality improver of gluten-free food formulated using hydrothermally treated polysaccharide mixtures (HTT-PSM) of glutinous rice flour and xanthan gum at different concentrations. The pasting and viscoelastic properties were determined by rapid visco analyser and rheometer respectively. The results showed that HTT-PSM had a lower gelatinisation temperature, higher peak viscosity and weaker gel strength than native glutinous rice flour. The incorporation of HTT-PSM into rice dough decreased the dough development time and stability significantly; and increased its extensibility from 2.8 mm to 11.9 mm. Its high extensibility attribute enabled the dough to make gluten-free noodles with higher tensile strength and similar texture profile as compared to wheat noodles. Sensory evaluation showed that the overall texture of the gluten-free noodles was acceptable, but not as comparable to wheat noodles. Positive correlations were observed between xanthan gum content and peak viscosity, gel strength, noodle tensile strength, hardness and chewiness.

The c-Jun N-terminal kinases, JNK1/2 are members of the mitogen activated protein kinase (MAPK) family that can mediate inflammatory responses, cell differentiation, cell proliferation, and/or cell death. In breast cancer, JNK signaling has been associated with increased proliferation, angiogenesis, and tumor metastasis. Also, inhibition of JNK has been shown to promote tumor cytostasis, but this inhibition also attenuates the cytotoxic effects of certain chemotherapeutics, calling into question the therapeutic potential of JNK inhibitors. In this study, we hypothesized that JNK1/2 inhibition induced autophagy, as a mechanism of protection against chemotherapy- and endocrine-induced apoptosis. Autophagy is a normal physiologic process required for the elimination of damaged mitochondria and misfolded proteins that can be induced in cancer cells under certain stresses such as hypoxia and drug insult to provide protection from death. To characterize the role of JNK1/2 in autophagy, we utilized two ERα expressing human breast cancer cell lines, T47-D and MCF-7. Autophagy flux experiments were performed in which the steady state levels of the autophagy protein LC3 II were analyzed in control cells versus cells undergoing JNK1/2 inhibition with the non-reversible JNK-IN8 inhibitor. JNK-IN8 was used as a single agent or in combination with estradiol and/or antiestrogen treatment. These studies reproducibly identified JNK1/2 inhibition as a mechanism of autophagy activation. The induction of autophagy correlated with sustained inhibition of proliferation as determined by cell counts, MTT, and clonogenic assays, and induction of senescence (in T47-D cells) as determined by the expression of β-galactosidase. In contrast, induction of apoptosis was not a major outcome of JNK1/2 inhibition. These pre-clinical studies provide strong evidence that JNK1/2 is an attractive molecular target to improve the treatment of endocrine responsive breast cancer, but they also emphasize that cells are surviving by autophagy and/or senescence. Because cell survival by autophagy and/or senescence could ultimately lead to breast cancer relapse, ongoing studies are addressing approaches to effectively kill these surviving cell populations. Citation Format: Kyler Herrington, Carol Joseph, Samar Abdulmoneim, Allison Lewis, Jingwen Cai, Hannah Youngblood, Yutao Liu, Patricia Schoenlein. Targeting JNK1/2 induces a potent cytostatic response in estrogen receptor expressing breast cancer cells, with induction of autophagy and senescence as measurable outcomes [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 1231.

Purpose Due to the limited availability of primary human Schlemm's canal (SC) endothelial cells, we aimed to develop an in vitro cellular model using the angular aqueous plexus (AAP) cells from bovine eyes. Methods We harvested a mixture of cells from the trabecular meshwork region including AAP loops from multiple donors, followed by puromycin treatment and immunostaining of Von Willebrand factor and vascular endothelial (VE)-cadherin to confirm identity. Previously identified differentially expressed genes in glaucomatous SC cells were examined in non-glaucomatous SC cells (n = 3) under 0% or 15% equibiaxial strain for 24 hours using droplet digital polymerase chain reaction (ddPCR) and analyzed using the Ingenuity Pathway Analysis (IPA) software application to identify upstream regulators. To compare the cellular responses to candidate regulators of these mechanoresponsive genes, AAP and human SC cells (n = 3) were treated with 5 or 10 ng/mL transforming growth factor beta-2 (TGFβ2) for 24 or 48 hours, followed with expression profiling using real-time PCR or ddPCR. Results We found that the isolated AAP cells displayed uniform cobblestone-like morphology and positive expression of two endothelial markers. In stretched SC cells, nine glaucoma-related genes were upregulated, and IPA implicated TGFβ as a potential upstream regulator. The effects of TGFβ2 treatment were similar for both AAP and SC cells in a dose- and time-dependent manner, activating TGFBR1 and SMAD2, inhibiting BMP4, and altering expression of three glaucoma-related genes (DCN, EZR, and CYP1B1). Conclusions Bovine AAP cells may serve as an alternative cellular model of human SC cells. Translational relevance These AAP cells may be used to study the functional pathways related to the outflow facility.

PurposeRegulation of intraocular pressure is dependent upon homeostatic responses of trabecular meshwork (TM) cells to mechanical stretch. Despite the important roles of miRNAs in regulating TM function and aqueous outflow, it remains unclear how miRNA and their target genes interact in response to physiological cyclic mechanical stretch. We aimed to identify differentially expressed miRNAs and their potential targets in human TM cells in response to cyclic mechanical stress.MethodsMonolayers of TM cells from non-glaucomatous donors (n=3-6) were cultured in the presence or absence of 15% mechanical stretch, 1 cycle/s, for 6 or 24-hours using computer-controlled Flexcell Unit. We profiled the expression of 800 miRNAs using NanoString Human miRNA assays and identified differentially expressed miRNAs using the Bioconductor Limma package. We identified differentially expressed genes using Operon Human Oligo Arrays with GeneSpring software. Pathway analysis with WebGestalt identified stretch-related pathways. We used Integrative miRNA Target Finder from Ingenuity Pathway Analysis to identify potential miRNA-mRNA regulations.ResultsWe identified 540 unique genes and 74 miRNAs with differential expression in TM cells upon cyclic mechanical stretch. Pathway analysis indicated the significant enrichment of genes involved in Wnt-signaling, receptor protein serine/threonine kinase signaling, TGF-β pathway, and response to unfolded protein. We also identified several miRNA master regulators, including miR-19b-3p and miR-93-5p, which may act as switches to control several mechano-responsive genes.ConclusionsThis study suggests that cyclic mechanical stress of TM cells triggers alterations in the expression of both mRNAs and miRNAs implicated in glaucoma-associated pathways.