Strategies that regulate LSD1 for novel therapeutics.

Histone methylation plays crucial roles in regulating chromatin structure and gene transcription in epigenetic modifications. Lysine-specific demethylase 1 (LSD1), the first identified histone demethylase, is universally overexpressed in various diseases. LSD1 dysregulation is closely associated with cancer, viral infections, and neurodegenerative diseases, etc., making it a promising therapeutic target. Several LSD1 inhibitors and two small-molecule degraders (UM171 and BEA-17) have entered the clinical stage. LSD1 can remove methyl groups from histone 3 at lysine 4 or lysine 9 (H3K4 or H3K9), resulting in either transcription repression or activation. While the roles of LSD1 in transcriptional regulation are well-established, studies have revealed that LSD1 can also be dynamically regulated by other factors. For example, the expression or activity of LSD1 can be regulated by many proteins that form transcriptional corepressor complexes with LSD1. Moreover, some post-transcriptional modifications and cellular metabolites can also regulate LSD1 expression or its demethylase activity. Therefore, in this review, we will systematically summarize how proteins involved in the transcriptional corepressor complex, various post-translational modifications, and metabolites act as regulatory factors for LSD1 activity.

Proapoptotic effect of WS-299 induced by NOXA accumulation and NRF2-counterbalanced oxidative stress damage through targeting RBX1-UBE2M interaction in gastric cancers.

The abnormal activation of Cullin RING E3 Ligases (CRLs) is closely associated with the occurrence and development of various cancers. Targeting the neddylation pathway represents an effective approach for cancer treatment. In this work, we reported that WS-299, structurally featuring a coumarin moiety attached to the triazolopyrimidine, exhibited excellent anti-proliferative activity in MGC-803 and HGC-27 cells. WS-299 exerted potent anticancer effects by inhibiting clone formation, EdU incorporation and inducing cell cycle arrest. WS-299 inhibited CUL3/5 neddylation and caused an obvious accumulation of Nrf2 and NOXA, substrates of CRL3 and CRL5, respectively. Biochemical studies showed that WS-299 inhibited CUL3 neddylation by inhibiting RBX1-UBE2M interaction. The anti-proliferative effect of WS-299 was mainly induced by NOXA-mediated apoptosis. Of note, Nrf2 attenuated WS-299-induced reactive oxygen species (ROS) levels. Furthermore, Nrf2 accumulation also had an antagonistic effect on NOXA-induced apoptosis. Therefore, WS-299 and siNrf2 synergistically increased ROS levels, apoptotic cells and suppressed tumor growth in vivo. Taken together, our research clarified the anti-cancer mechanisms of WS-299 through targeting the RBX1-UBE2M protein-protein interaction and inhibiting the neddylation modification of CUL3 and CUL5. More importantly, our studies also demonstrated that combination of WS-299 with shNrf2 could be an effective strategy for treating gastric cancers.

VEGF165b Mutant Promotes the Apoptosis of Murine Breast Cancer Cells Induced by Paclitaxel by Inducing Tumor Vessel Maturation.

INTRODUCTION: The anti-angiogenic agent vascular endothelial growth factor 165b (VEGF165b) mutant (mVEGF165b), which was developed by our laboratory, has superior antitumor activity to that of native VEGF165b; however, its mechanism of action and druggability need further exploration. METHODS: Using the commercial anti-angiogenic drug bevacizumab as a positive control, the mechanism and developability of mVEGF165b were evaluated and explored. The Cell Counting Kit-8 assay was performed to evaluate the effects of mVEGF165b and bevacizumab alone on the proliferation of human umbilical vein endothelial cells (HUVECs). Meanwhile, the inhibitory effects of mVEGF165b and bevacizumab combined with paclitaxel in a mouse model of breast cancer were assessed. Immunohistochemistry was used to detect their effects on tumor vascular maturation, and the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay was used to detect the apoptosis of tumor cells. RESULTS: In vitro cell experiments confirmed that mVEGF165b inhibited the proliferation of HUVECs with an efficacy equivalent to that of bevacizumab. mVEGF165b and bevacizumab combined with paclitaxel significantly delayed the growth of breast cancer in mice. Immunohistochemistry and the TUNEL assay showed that mVEGF165b and bevacizumab combined with paclitaxel-induced higher vascular maturity and more apoptosis than paclitaxel alone. CONCLUSION: mVEGF165b showed similar efficacy and mechanism of action as bevacizumab, indicating its potential to be developed into a safe and effective anti-angiogenic drug.

Novel nitrogen removal process in marine aquaculture wastewater treatment using Enteromorpha ferment liquid as carbon.

The process performance of partial denitrification of a novel anaerobic fermentation integrated fixed-film activated sludge (IFAS-AFPD) of Enteromorpha was studied. The response surface method was used to determine the optimal reaction conditions, and the operation experiment was carried out under the optimal conditions. The results showed that the nitrogen removal effect was the best when the salinity was 12.2 g•L-1, the Carbon-Nitrogen ratio (C/N) was 4, the pH was 8.5, and the Nitrite Accumulation Rate, Nitrate Removal Rate, Chemical Oxygen Demand Utilization Rate could reach 77%, 89% and 51%. Experimental results have shown that the NAR of the Enteromorpha ferment liquid system could be maintained at about 74%, which was noteworthy higher than that of the sodium acetate (CH3COONa) system at 42%; Microbial community analysis showed that Enteromorpha ferment liquid was more beneficial to the growth of Bacteroidetes than CH3COONa.

Leiomyoma with Bizarre Nuclei: A Current Update.

Leiomyoma with bizarre nuclei (LBN), also known as symplastic leiomyoma, is a histological subtype of benign leiomyoma with bizarre cells and nuclear atypia. Differentiating LBN from other benign leiomyoma subtypes, uterine smooth muscle tumors of uncertain malignant potential (STUMP), or leiomyosarcoma (LMS) can be diagnostically challenging owing to overlapping features in clinical presentation and pathologic morphological analysis. The difficulty of distinguishing LBN from other lesions, especially from LMS, and the potential of LBN for subsequent malignant transformation make LBN an important topic of research. Herein, we review the definition, diagnosis, treatment, and prognosis of LBN. Histopathological examination is essential for distinguishing LBN from other diseases. Pathology sampling and morphological examination remain the key to diagnosis. The newly established ancillary immunohistochemical (IHC) and molecular genetic analysis can be useful tools for differential diagnosis. Furthermore, serum biomarkers and imaging examination may also be useful diagnostic tools. Attention should be paid to the differentiation between LBN and LMS because morphological diagnosis may still be challenging in some cases. Some IHC markers of LBN have been identified, which may be helpful for differential diagnosis. Furthermore, the use of IHC panels as diagnostic markers may be advocated. Molecular genetic studies suggest that some genes can aid with the differential diagnosis between LBN and LMS. However, increasing evidence support the idea that LBN and LMS are molecularly related, indicating that LBN may represent a potentially malignant stage of precancerous progression. At present, conservative treatment is recommended for primary LBN, especially for patients desiring to retain fertility, but close follow-up with imaging examinations is required.

Quantitative and ultrasensitive detection of brombuterol by a surface-enhanced Raman scattering (SERS)-based lateral flow immunochromatographic assay (FLIA) using AgMBA@Au-Ab as an immunoprobe.

Brombuterol is a new emerging ß-adrenergic agonist that has been used as an additive in animal feed to enhance the lean meat-to-fat ratio. Due to its potential harm to consumers, it is urgent to develop sensitive, simple and rapid analytical methods to monitor brombuterol residue. In this study, a competitive lateral flow immunochromatographic assay (FLIA) based on surface-enhanced Raman scattering (SERS) was developed for ultrasensitive quantitative determination of brombuterol in swine liver, pork and feed samples. Ag@Au core-shell bimetallic nanoparticles with the highest SERS enhancement were synthesized, characterized and used as the substrate for preparation of the immunoprobe AgMBA@Au-Ab, in which the Raman reporter mercaptobenzoic acid (MBA) was embedded between the core-shell layers and monoclonal antibodies against brombuterol were immobilized on the surfaces of nanoparticles. The presence of brombuterol was identified through a color change on testing lines. In addition, quantitative detection of brombuterol was achieved by measuring the characteristic Raman peak intensity of MBA in the immunoprobes captured by the coating antigen. The IC50 and limit of detection (LOD) of the SERS-based FLIA for brombuterol were 45 pg mL-1 and 0.11 pg mL-1, respectively. The recoveries of brombuterol from spiked samples were in the range of 87.27-100.16% with relative standard deviations of 1.29%-6.99% (n = 3). The proposed SERS-based LFIA was proven to be a feasible method for ultrasensitive and rapid detection of brombuterol and might be a platform for sensitive and rapid detection of a broad range of analytes in clinical, environmental and food analyses.

Ischaemic stroke etiological classification system: the agreement analysis of CISS, SPARKLE and TOAST.

BACKGROUND AND PURPOSE: The ideal stroke classification system needs to have validity, high reliability and applicability among different stroke research settings. The Chinese Ischemic Stroke Subclassification (CISS) and the Subtypes of Ischemic Stroke Classification System (SPARKLE) have emerged recently but have not been tested using agreement analysis. As a result, the objective of this study is to investigate the level of agreement among stroke subtype classifications using CISS, SPARKLE and Trial of Org 10172 in Acute Stroke Treatment (TOAST). We also analyse the inter-rater reliability of CISS. METHODS: The data include 623 inpatients who have had an ischaemic stroke, accrued from Beijing Tiantan Hospital between 1 October 2015 and 19 April 2016. According to the diagnostic standards of the three subtype classification systems, 299 inpatients who satisfied the requirements of our study were independently classified with etiological subtypes, and we compared the three subclassifications. RESULTS: There was substantial overall agreement among the three classification systems: CISS versus SPARKLE (kappa value=0.684, p<0.001), CISS versus TOAST (kappa value=0.615, p<0.001) and SPARKLE versus TOAST (kappa value=0.675, p<0.001). The inter-rater reliability of CISS was excellent (kappa value=0.857, p<0.001). Furthermore, among the three subtype classification systems, the variance analysis results of the etiological subtypes were not uniform. CONCLUSION: There were generally substantial agreements among three ischaemic stroke etiological classification systems. CISS is a valid and reliable classification system, with which different stroke research centres can apply and compare data.

Efficient synthesis of riboflavin-imprinted magnetic nanoparticles by boronate affinity-based surface imprinting for the selective recognition of riboflavin.

Riboflavin (vitamin B2), a cis-diol-containing compound, is an essential vitamin for maintaining human health mainly in energy metabolism and is a critical component of enzyme cofactors and flavoproteins. Thus, the determination of riboflavin in food products is essential in riboflavin-fortified foods. However, analysis of riboflavin at natural levels in foods or biological samples is difficult because of its very low concentration level and the presence of undesirable matrix effects which could interfere with the measuring system. Thus, it is essential to develop efficient and selective enrichment approaches for riboflavin. Molecularly imprinted polymers can be well used for riboflavin extraction and pre-concentration from food samples. In this study, we present riboflavin-imprinted magnetic nanoparticles as an ideal sorbent for the selective enrichment of riboflavin followed by direct fluorometric determination. The riboflavin-imprinted magnetic nanoparticles were prepared according to a newly reported strategy, called boronate affinity-based surface initiated imprinting. Magnetic nanoparticles and vinylphenylboronic acid were used as supporting materials and a functional monomer, respectively. The prepared riboflavin-imprinted magnetic nanoparticles exhibited several significant advantages, such as excellent selectivity, high binding affinity and low binding pH toward riboflavin, which made the molecularly imprinted material become an ideal sorbent for the selective enrichment of riboflavin. The prepared riboflavin-imprinted magnetic nanoparticles were successfully applied to the analysis of riboflavin in milk.

Synthesis and QSAR study of novel α-methylene-γ-butyrolactone derivatives as antifungal agents.

Thirty-six new α-benzylidene-γ-lactone compounds based α-methylene-γ-butyrolactone substructure were prepared and characterized by spectroscopic analysis. All compounds were evaluated for antifungal activities in vitro against six plant pathogenic fungi and the half maximal inhibitory concentration (IC50) against Botrytis cinerea and Colletotrichum lagenarium were investigated. Compounds 5c-3 and 5c-5 with the halogen atom exhibited excellent fungicidal activity against B. cinerea (IC50=22.91, 18.89µM). The structure-activity relationships (SARs) analysis indicated that the derivatives with electron-withdrawing substituents at the meta- or para-positions improves the activity. Via the heuristic method, the generated quantitative structure-activity relationship (QSAR) model (R2=0.961) revealed a strong correlation of antifungal activity against B. cinerea with molecular structures of these compounds. Meanwhile, the cytotoxicity of 20 representative derivatives was tested in the human tumor cells line (HepG2) and the hepatic L02 cells line, the result indicated that the synthesized compounds showed significant inhibitory activity and limited selectivity. Compound 5c-5 has the highest fungicidal activity with IC50=18.89µM (against B. cinerea.) but low cytotoxicity with IC50=35.4µM (against HepG2 cell line) and IC50=68.8µM (against Hepatic L02 cell line). These encouraging results can be providing an alternative, promising use of α-benzylidene-γ-lactone through the design and exploration of eco-friendly fungicides with low toxicity and high efficiency.