Repurposing diacerein to suppress colorectal cancer growth by inhibiting the DCLK1/STAT3 signaling pathway.

Double cortin-like kinase 1 (DCLK1) exhibits high expression levels across various cancers, notably in human colorectal cancer (CRC). Diacerein, a clinically approved interleukin (IL)-1ß inhibitor for osteoarthritis treatment, was evaluated for its impact on CRC proliferation and migration, alongside its underlying mechanisms, through both in vitro and in vivo analyses. The study employed MTT assay, colony formation, wound healing, transwell assays, flow cytometry, and Hoechst 33342 staining to assess cell proliferation, migration, and apoptosis. Additionally, proteome microarray assay and western blotting analyses were conducted to elucidate diacerein's specific mechanism of action. Our findings indicate that diacerein significantly inhibits DCLK1-dependent CRC growth in vitro and in vivo. Through high-throughput proteomics microarray and molecular docking studies, we identified that diacerein directly interacts with DCLK1. Mechanistically, the suppression of p-STAT3 expression following DCLK1 inhibition by diacerein or specific DCLK1 siRNA was observed. Furthermore, diacerein effectively disrupted the DCLK1/STAT3 signaling pathway and its downstream targets, including MCL-1, VEGF, and survivin, thereby inhibiting CRC progression in a mouse model, thereby inhibiting CRC progression in a mouse model.

Feeding ecology of the short-headed lanternfish Diaphus brachycephalus and Warming's lanternfish Ceratoscopelus warmingii (Myctophidae) in the South China Sea.

We report on the feeding ecology of two species, the short-headed lanternfish Diaphus brachycephalus and Warming's lanternfish Ceratoscopelus warmingii, using data collected over five surveys from 2015 to 2017 in the open South China Sea. D. brachycephalus feed mainly on copepods, with few differences in food composition between different-sized individuals; the diet of C. warmingii is more diverse, including crustacean zooplankton, gelatinous animals, and Mollusca, and differs significantly between fishes >55 mm in body length and smaller fishes. Interspecific competition for food between these two species is not strong, while intraspecific competition may be more intense in D. brachycephalus than in C. warmingii. Trophic levels of D. brachycephalus (3.46) and C. warmingii (3.38) identify both species as third-trophic-level lower carnivores. The diel feeding patterns of D. brachycephalus and C. warmingii differ: the former feeds actively both day and night when food is plentiful, and feeds primarily in the upper layer at night and in the mesopelagic layer during the daytime, and the latter ascends into the upper 100 m at night to feed, but stomach fullness is lower than D. brachycephalus. Dry-body-weight daily ration estimates for D. brachycephalus range from 5.19% to 16.46%, and those for C. warmingii range from 1.38% to 4.39%.

Stable isotopic and stomach content analyses reveal changes in the trophic level and feeding habit of large-head hairtail (Trichiurus lepturus) in the northern South China Sea.

The feeding habit of large-head hairtail (Trichiurus lepturus) in the northern South China Sea was investigated through isotopic and stomach content analyses. The isotopic features of the hairtail at the same body size differed among regions, with the fish in coastal waters presenting higher δ15N and δ13C values compared to those in the open sea, indicating different trophic levels (TL), food habits, and isotopic baselines. According to the partial correlation of water depth with δ15N values, the sampling stations were divided into three regions based on the depth of water: coastal (20-40 m), near coastal (60-80 m), and open sea (100-200 m) regions. In the coastal region, the hairtail from stations affected by the Pearl River plume exhibited lower δ15N and δ13C values. The stomach content analysis indicated different feeding habits of the hairtail from different regions. The hairtail in the coastal and near coastal waters fed more on fish and less on crustaceans compared to the hairtail in the open sea. The relationship between δ15N and fish size exhibited two contrary patterns. First, the δ15N values increased with increasing preanal length in the hairtail sampled from the water depth of 30-40 m in section F (in fish with preanal length < 200 mm) and those samples from the water depth of 100-200 m. This finding reflected an ontogenetic shift in diet and TL. However, the δ15N values tended to decrease with the increasing preanal length of the hairtail samples collected from the water depth of 30-40 m in section F (fish with a preanal length of ~200-300 mm). These findings suggested that under the conditions of insufficient availability of high-quality prey, the larger hairtail fed more on low-TL prey to compensate for the increase energy demand, arising due to growth, which led to the observed decrease in δ15N values.

Siting MPAs for multiple protecting purposes by co-consideration of ecological importance and anthropogenic impacts.

The global marine ecosystem has been significantly altered by the combined effects of multiple anthropogenic impacts. Systematic planning of marine protected areas (MPAs) is of paramount importance in alleviating conflicts between humans and the sea. Existing approaches, however, merely integrate both ecological and anthropogenic factors for multiple conservation purposes. By combining the three main anthropogenic impact factors with two main ecological importance factors, this study used a GIS-based AHP-OWA method to identify different levels of priority protection for MPAs in Zhejiang, China. Our results proved that: 1) the multi-objective MPA siting issues can be addressed by the GIS-based AHP-OWA method through scenario simulation; 2) the best locations for MPAs are in the northeast, central, and southern marine areas of Zhejiang; 3) considering the trade-off degree, spatial conservation efficiency, and spatial heterogeneity, an optimized MPA siting scheme can be developed for decision-makers. The proposed MPA siting method and case study may provide an effective technical reference for solving regional marine spatial planning (MSP) issues in the future.

Pharmacological targeting of MTHFD2 suppresses NSCLC via the regulation of ILK signaling pathway.

Lung cancer is the most common cause of cancer related deaths worldwide with the highest mortality rate. Non-small cell lung cancer (NSCLC) accounts for about 85 % of lung cancers. Mitochondrial methylenetetrahydrofolate dehydrogenase 2 (MTHFD2) is a bifunctional enzyme and is the most differentially expressed metabolic enzyme in various tumors including lung cancer. However, little is known about how MTHFD2 functions in NSCLC. Integrin-linked kinase (ILK) signaling plays key a role in tumor progression including metastasis, proliferation and migration. Here, we show that MTHFD2 inhibition results in suppression of cell growth, migration, invasion and epithelial-mesenchymal transition (EMT) in NSCLC. Microarray analysis suggests that MTHFD2 is positively associated with ILK signaling based on western blotting results. In addition, the phosphorylation of AMPKα plays an essential role in MTHFD2 regulation of ILK signaling. Further, the small-molecule compound C18 inhibits MTHFD2 with great efficiency. C18 blocks MTHFD2/ILK signaling pathway and restrains cell growth, migration, invasion, and EMT of NSCLC and induces apoptosis. In brief, our study found that the positive impact of MTHFD2 is mediated via ILK signaling pathway in NSCLC. Thus, blocking MTHFD2 represents a promising therapeutic strategy against NSCLC clinically.

Celastrol elicits antitumor effects by inhibiting the STAT3 pathway through ROS accumulation in non-small cell lung cancer.

BACKGROUND: Non-small cell lung cancer (NSCLC) is the most common lung cancer with high mortality across the world, but it is challenging to develop an effective therapy for NSCLC. Celastrol is a natural bioactive compound, which has been found to possess potential antitumor activity. However, the underlying molecular mechanisms of celastrol activity in NSCLC remain elusive. METHODS: Cellular function assays were performed to study the suppressive role of celastrol in human NSCLC cells (H460, PC-9, and H520) and human bronchial epithelial cells BEAS-2B. Cell apoptosis levels were analyzed by flow cytometry, Hoechst 33342, caspase-3 activity analysis, and western blot analysis. Intracellular reactive oxygen species (ROS) were analyzed by flow cytometry and fluorescence microscope. Expression levels of endoplasmic reticulum (ER) stress-related proteins and phosphorylated signal transducer and activator of transcription 3 (P-STAT3) were identified via western blot analysis. A heterograft model in nude mice was employed to evaluate the effect of celastrol in vivo. RESULTS: Celastrol suppressed the growth, proliferation, and metastasis of NSCLC cells. Celastrol significantly increased the level of intracellular ROS; thus, triggering the activation of the ER stress pathway and inhibition of the P-STAT3 pathway, and eventually leading to cell apoptosis, and the effects were reversed by the pre-treatment with N-Acetyl-L-cysteine (NAC). Celastrol also suppressed tumor growth in vivo. CONCLUSION: The outcomes revealed that celastrol plays a potent suppressive role in NSCLC in vitro and in vivo. Celastrol induces apoptosis via causing mitochondrial ROS accumulation to suppress the STAT3 pathway. Celastrol may have potential application prospects in the therapy of NSCLC.

Low-depth whole genome sequencing reveals copy number variations associated with higher pathologic grading and more aggressive subtypes of lung non-mucinous adenocarcinoma.

OBJECTIVE: Histology grade, subtypes and TNM stage of lung adenocarcinomas are useful predictors of prognosis and survival. The aim of the study was to investigate the relationship between chromosomal instability, morphological subtypes and the grading system used in lung non-mucinous adenocarcinoma (LNMA). METHODS: We developed a whole genome copy number variation (WGCNV) scoring system and applied next generation sequencing to evaluate CNVs present in 91 LNMA tumor samples. RESULTS: Higher histological grades, aggressive subtypes and more advanced TNM staging were associated with an increased WGCNV score, particularly in CNV regions enriched for tumor suppressor genes and oncogenes. In addition, we demonstrate that 24-chromosome CNV profiling can be performed reliably from specific cell types (<100 cells) isolated by sample laser capture microdissection. CONCLUSIONS: Our findings suggest that the WGCNV scoring system we developed may have potential value as an adjunct test for predicting the prognosis of patients diagnosed with LNMA.

Clinical features and prognostic factors for surgical treatment of esophageal squamous cell carcinoma in elderly patients.

PURPOSE: This research was designed to analyze the clinical features and prognostic factors for surgical treatment of esophageal squamous cell carcinoma (ESCC) in elderly patients aged 70 years and over. METHODS: The clinical data and follow-up data of 68 ESCC patients aged 70 years and over were collected. The characteristics of surgical treatment, perioperative complications, overall survival (OS), and the factors affecting survival were analyzed. RESULTS: The incidence rate of postoperative complications was 36% and the mortality rate was 0% during and 90 days after surgery. The 5-year OS was 45.0% and the 5-year disease-free survival (DFS) was 38.0%. Univariate analysis showed that gender, Charlson Comorbidity Index (CCI), pathological type, tumor differentiation, depth of invasion, postoperative complications, and lymph node metastasis were the factors associated with OS. Multivariate analysis showed that pathologic type, depth of invasion, and lymph node metastasis were the independent predictors of OS. The ideal long-term survival in elderly patients with ESCC was achieved with radical resection. CONCLUSION: The pathological type and pathological stage were the important independent risk factors of prognosis.

A comprehensive multiplex PCR based exome-sequencing assay for rapid bloodspot confirmation of inborn errors of metabolism.

BACKGROUND: Tandem mass spectrometry (MS MS) and simple fluorometric assays are currently used in newborn screening programs to detect inborn errors of metabolism (IEM). The aim of the study was to evaluate the clinical utility of exome sequencing as a second tier screening method to assist clinical diagnosis of the newborn. METHODS: A novel PCR-exome amplification and re-sequencing (PEARS) assay was designed and used to detect mutations in 122 genes associated with 101 IEM. Newborn bloodspots positive by biochemical testing were analysed by PEARS assay to detect pathogenic mutations relevant to the IEM. RESULTS: In initial validation studies of genomic DNA samples, PEARS assay correctly detected 25 known mutations associated with 17 different IEM. Retrospective gene analysis of newborns with clinical phenylketonuria (PKU), identified compound heterozygote phenylalanine hydroxylase (PAH) gene mutations in eight of nine samples (89%). Prospective analysis of 211 bloodspots correctly identified the two true PKU samples, yielding positive and negative predictive values of 100%. Testing of 8 true positive MS MS samples correctly identified potentially pathogenic compound heterozygote genotypes in 2 cases of citrullinemia type 1 and one case each of methylmalonic acidemia, isobutyryl-CoA dehydrogenase deficiency, short chain acyl-CoA dehydrogenase deficiency and glutaric acid type II and heterozygous genotypes in 2 cases of autosomal dominant methioninemia. Analysis of 11 of 12 false positive MS MS samples for other IEM identified heterozygous carriers in 8 cases for the relevant genes associated with the suspected IEM. In the remaining 3 cases, the test revealed compound heterozygote mutations in other metabolic genes not associated with the suspected IEM, indicating a misinterpretation of the original MS MS data. CONCLUSIONS: The PEARS assay has clinical utility as a rapid and cost effective second-tier test to assist the clinician to accurately diagnose newborns with a suspected IEM.

Clinical application of single-molecule optical mapping to a multigeneration FSHD1 pedigree.

INTRODUCTION: Facioscapulohumeral muscular dystrophy 1 (FSHD1) is a relatively common autosomal dominant adult muscular dystrophy with variable disease penetrance. The disease is caused by shortening of a D4Z4 repeat array located near the telomere of chromosome 4 at 4q35. This causes activation of a dormant gene DUX4, permitting aberrant DUX4 expression which is toxic to muscles. Molecular diagnosis of FSHD1 by Southern blot hybridization or FISH combing is difficult and time consuming, requiring specialist laboratories. As an alternative, we apply a novel approach for the diagnosis of FSHD1 utilizing single-molecule optical mapping (SMOM). METHODS: Long DNA molecules with BssS1 enzyme marking were subjected to SMOM on the Bionano Genomics platform to determine the number of D4Z4 repeats. Southern blot and molecular combing were used to confirm the FSHD1 haplotypes. RESULTS: In a study of a five-generation FSHD1 pedigree, SMOM correctly diagnosed the disease and normal haplotypes, identifying the founder 4qA disease allele as having 4 D4Z4 repeat units. Southern blot and molecular combing analysis confirmed the SMOM results for the 4qA disease and 4qB nondisease alleles. CONCLUSION: Based on our findings, we propose that SMOM is a reliable and accurate technique suitable for the molecular diagnosis of FSHD1.

Twisted palladium-copper nanochains toward efficient electrocatalytic oxidation of formic acid.

Twisted PdCu nanochains are synthesized successfully via a staged thermal treatment route, offering rich twin boundaries as catalytic "active sites" and modified electronic effects. Toward formic acid oxidation, the twisted PdCu nanochains hold the highest catalytic peak current density (1108.2 mA mg-1Pd) over previous reported PdCu alloy catalysts, and also much higher catalytic activity and durability comparing with Pd nanochains and commercial Pd/C. The catalytic enhancement mechanism to PdCu nanochains is proposed and discussed. Additionally, we found that the formation of PdCu nanochains follows a typical anisotropic growth approach, and the multiple steps of staged thermal treatment route displays a vital role in fabricating the unique PdCu nanochains while the introduced Cu precursors might affect the reduction rate of Pd species and act as deposition or nucleation sites for twisted structure in terms of rich twin boundaries. This work describes an efficient, low-Pd loading catalyst for electrooxidation of formic acid, and also demonstrates a universal method to fabricate other defect-rich catalysts for broad applications in energy conversion and storage systems and sensing devices.

Synthesis of defect-rich palladium-tin alloy nanochain networks for formic acid oxidation.

Unique and novel Pd4Sn nanochain networks were successfully synthesized with an average diameter of 5 nm, rendering a modified Pd electronic structure with rich defects such as atomic corners, steps or ledges as catalytic active sites for great enhancement of charge transfer and electrode kinetics. The prepared Pd4Sn nanochain networks held an electrochemically active surface area as high as 119.40 m2 g-1, and exhibited higher catalytic activity and stability toward formic acid oxidation compared with Pd3Sn nanochain networks, Pd5Sn nanochain networks, Pd4Sn dendrites and Pd/C. The fundamental insight of the enhancement mechanism is discussed, and this work offers a novel, less expensive but highly active catalyst for direct formic acid fuel cells.

Involvement of Ras GTPase-activating protein SH3 domain-binding protein 1 in the epithelial-to-mesenchymal transition-induced metastasis of breast cancer cells via the Smad signaling pathway.

In situ models of epithelial-to-mesenchymal transition (EMT)-induced carcinoma develop into metastatic carcinoma, which is associated with drug resistance and disease recurrence in human breast cancer. Ras GTPase-activating protein SH3 domain-binding protein 1 (G3BP1), an essential Ras mediator, has been implicated in cancer development, including cell growth, motility, invasion and apoptosis. Here, we demonstrated that the upregulation of G3BP1 activates the EMT in breast cancer cells. Silencing Smads almost completely blocked this G3BP1-induced EMT, suggesting that this process depends on the Smad signaling pathway. We also found that G3BP1 interacted with the Smad complex. Based on these results, we proposed that G3BP1 might act as a novel co-factor of Smads by regulating their phosphorylation status. Moreover, knockdown of G3BP1 suppressed the mesenchymal phenotype of MDA-MB-231 cells in vitro and suppressed tumor growth and lung metastasis of 4T1 cells in vivo. Our findings identified a novel function of G3BP1 in the progression of breast cancer via activation of the EMT, indicating that G3BP1 might represent a potential therapeutic target for metastatic human breast cancer.

Phosphorylation of myofibrillogenesis regulator-1 activates the MAPK signaling pathway and induces proliferation and migration in human breast cancer MCF7 cells.

Myofibrillogenesis regulator-1 (MR-1) has been characterized as a tumor promoter in many cancers. However, its mechanism of action has not been fully elucidated. Here, we report that MR-1 is overexpressed in human breast cancer cells and participates in tumor promotion in human breast cancer MCF7 cells by activating the ERK1/2 signaling pathway. MR-1 interacts with MEK1/2 and ERK1, and its N-terminal sequence plays a major role in promoting the MEK/ERK cascade. Furthermore, six phosphorylation sites of MR-1 were identified, and phosphorylation at S46 was shown to be critical for the activation of MEK/ERK. Therefore, our findings suggest that MR-1 functions as a tumor promoter in MCF7 cells by activating the MEK/ERK signaling.

[Inhibitions of SphK1 inhibitor SKI II on cell cycle progression and cell invasion of hepatoma HepG2 cells].

Sphingosine kinase 1 (SphK1) plays critical roles in cell biological functions. Here we investigated the effects of SphK1 inhibitor SKI II on hepatoma HepG2 cell cycle progression and invasion. Cell survival was determined by SRB assay, cell cycle progression was assayed by flow cytometry, the ability of cell invasion was measured by Matrigel-Transwell assay and protein expression was detected by Western blotting. The results showed that SKI II markedly inhibited HepG2 cell survival in a dose-dependent manner, induced G1 phase arrest in HepG2 cell and inhibited cell invasion. SKI II markedly decreased the expressions of G1-phase-related proteins CDK2, CDK4 and Cdc2 and the levels of cell invasion-associated proteins MMP2 and MMP9. The results showed that SKI II inhibited cell cycle progression and cell invasion, implying SphK1 as a potential target for hepatoma treatment.