Iron 3D-Orbital Configuration Dependent Electron Transfer for Efficient Fenton-Like Catalysis.

Transition metals are excellent active sites to activate peroxymonosulfate (PMS) for water treatment, but the favorable electronic structures governing  reaction mechanism still remain elusive. Herein, the authors construct typical d-orbital configurations on iron octahedral (FeOh ) and tetrahedral (FeTd ) sites in spinel ZnFe2 O4 and FeAl2 O4 , respectively. ZnFe2 O4 (136.58 min-1 F-1 cm2 ) presented higher specific activity than FeAl2 O4 (97.47 min-1 F-1 cm2 ) for tetracycline removal by PMS activation. Considering orbital features of charge amount, spin state, and orbital arrangement by magnetic spectroscopic analysis, ZnFe2 O4 has a larger bond order to decompose PMS. Using this descriptor, high-spin FeOh is assumed to activate PMS mainly to produce nonradical reactive oxygen species (ROS) while high-spin FeTd prefers to induce radical species. This hypothesis is confirmed by the selective predominant ROS of 1 O2 on ZnFe2 O4 and O2 •- on FeAl2 O4 via quenching experiments. Electrochemical determinations reveal that FeOh has superior capability than FeTd for feasible valence transformation of iron cations and fast interfacial electron transfer. DFT calculations further suggest octahedral d-orbital configuration of ZnFe2 O4 is beneficial to enhancing Fe-O covalence for electron exchange. This work attempts to understand the d-orbital configuration-dependent PMS activation to design efficient catalysts.

Porous Carbon Nitride Frameworks Derived from Covalent Triazine Framework Anchored Ag Nanoparticles for Catalytic CO2 Conversion.

Porous carbon nitride frameworks (PCNFs) with uniform and rich nitrogen dopants and abundant porosity were successfully fabricated through the direct carbonization of the covalent triazine frameworks (CTFs) at different pyrolysis temperatures and used as supports to anchor and stabilize Ag nanoparticles (NPs) for catalytic CO2 conversion. Importantly, the pyrolysis temperature plays a crucial role in the properties of porous carbon nitride frameworks. The material carbonized at 700 °C showed the highest surface area and micro- and mesoporous structure with a certain interlayer distance. Taking advantage of their unique surface characteristics, PCNF-supported Ag NP catalysts (Ag/PCNF-T, T=pyrolysis temperature) were prepared by a simple chemical method. A series of characterizations revealed that Ag NPs are embedded in the porous carbon nitride frameworks and confined to a relatively small size with high dispersion owing to the assistance of the abundant surface groups and porous structures. The as-obtained Ag/PCNF-T catalysts, especially Ag/PCNF-700, showed excellent catalytic activity, selectivity, and stability for the carboxylation of CO2 with terminal alkynes under mild conditions. This can be due to the existence of abundant nitrogen atoms and diverse porosity, which resulted in highly efficient catalytic activity and stability.

Competitive light-initiated chemiluminescent assay: using 5-α-dihydrotestosterone-BSA as competitive antigen for quantitation of total testosterone in human sera.

This paper described a homogeneous method, light-initiated chemiluminescent assay (LICA), for quantitation of total testosterone in human sera. The assay was bead based and built on a competitive-binding reaction format, in which 5-α-dihydrotestosterone (5-α-DHT) competed with the testosterone in serum samples in binding with biotinylated anti-testosterone antibody. The more testosterone in the serum sample, the less 5-α-DHT that bonded with biotinylated anti-testosterone antibodies. 5-α-DHT was coupled with emission beads (doped with thioxene derivatives and Eu(III) as a chemiluminescence emitter) via bovine serum albumin as a linker. Once streptavidin-coated sensitizer beads (modified with phthalocyanine as a photosensitizer) were added, the streptavidin/biotin reaction between 5-α-DHT-bound anti-testosterone antibody and sensitizer beads could bring emission and sensitizer beads together, which allowed energy transfer from sensitizer bead to emission bead. As such, an exciting light (680 nm) impinging on the sensitizer beads led to light emission at 520-620 nm by emission beads. The strength of the emitted light was inversely proportional to the testosterone in serum sample. The detection range of this assay was from 13.3 to 1200 ng/dL. The coefficient variation for intra- and inter-assay was lower than 15%. The recovery of this method ranged from 95.5 to 105.9% for different samples. Moreover, the LICA assay was highly specific with low cross-reactivity and interference. The concentration of testosterone from 58 serum samples analyzed by the LICA method significantly correlated (y = 0.97x + 1.87, R2 = 0.970, p < 0.001) with those obtained with the SIEMENS Centaur Xp System. Graphical abstract ᅟ.

Development of a light-initiated chemiluminescent assay for the quantitation of sIgE against egg white allergens based on component-resolved diagnosis.

The determination of specific IgE (sIgE) level is of great importance in IgE-mediated food allergies. Our aim was to develop a homogeneous immunoassay-light-initiated chemiluminescent assay (LICA)-for measuring allergen sIgE of a single component in egg white, thus evaluating the LICA-sIgE assay as a useful tool in the diagnosis of food allergy. The LICA-sIgE assay was performed by incubating serum sample with anti-human IgE antibody coated with chemiluminescer beads, streptavidin-coated sensitizer beads, and biotinylated antigens, which consist of four components in egg white. Serum samples from egg allergic patients (n = 70) and healthy volunteers (n = 30) were collected. For calibration, purified human IgE was used as the calibrator. Working conditions of this homogeneous immunoassay were optimized, analytical performance was determined, and correlation of the results between LICA and ImmunoCAP was evaluated. The assays were performed in 8-well plates with a sample volume diluted to 1:10 of 25 µl. Intra-assay precision (% coefficient of variation) ranged from 1.83 to 4.13%, and inter-assay precision ranged from 2.70 to 8.70%. It exhibited excellent sensitivity, which could distinguish between positive samples and negative samples even at a large dilution level. The sIgE-LICA and ImmunoCAP correlated well in patients allergic to single component (r 2 = 0.929). Also, the components ovomucoid and ovalbumin were best at predicting ImmunoCAP results, with the same area under the ROC curve (AUC) of 0.81, and a specificity of 90.0 and 93.3%, respectively. Our data show effective performance characteristics of LICA to detect sIgE in human serum based on component-resolved diagnostic tests (CRD). The homogeneous sIgE-LICA assay has the following key advantages: requires no washing, simplicity and rapidity, reproducibility, high-throughput, good performance in a liquid phase assay, and good suitability for sIgE diagnosis in food allergy based on CRD. Graphical abstract A light-initiated chemiluminescent assay was developed for the quantitation of sIgE against egg white allergens based on component-resolved diagnosis. Components Gal d 1 and Gal d 2 with the highest AUC values of 0.81 were considered the best at predicting egg allergy.

Sarsaparilla (Smilax Glabra Rhizome) Extract Activates Redox-Dependent ATM/ATR Pathway to Inhibit Cancer Cell Growth by S Phase Arrest, Apoptosis, and Autophagy.

Sarsaparilla (Smilax Glabra Rhizome) exerts growth inhibitory effect on multiple cancer cells in vitro and in vivo, and redox-dependent persistent activation of ERK1/2 has been reported to underlie this effect. Here, we report an activation of ATM/ATR-dependent signaling pathway also as a mechanism for the cancer cell growth inhibition induced by the supernatant fraction of the water-soluble extract from sarsaparilla (SW). SW treatment (3.5 µg/µL) promoted the phosphorylations of ATM, ATR, and CHK1 in AGS and HT-29 cells. The ATM kinase inhibitor, KU55933, could reverse SW-induced ERK phosphorylation but not the reduced glutathione/oxidized glutathione (GSH/GSSG) imbalance in AGS cells. However, both the redox inhibitor glutathione (GSH) and ERK inhibitor U0126 antagonized SW-induced phosphorylations of ATM, ATR, and CHK1 in AGS cells. We further found KU55933 significantly antagonized SW-induced S phase arrest, apoptosis, autophagy and the resultant cell growth inhibition. Our results provide another molecular basis for the anticancer action of sarsaparilla.

Co-encapsulation of granzyme B and perforin in nanocapsules for tumour therapy: biomimicking immune cells.

Granzyme B (GrB)-based immunotherapy is of interest for cancer treatment. However, insufficient cellular uptake and a lack of targeting remain challenges to make use of GrB for solid tumour therapy. As GrB induced cell death requires the help of perforin (PFN), we designed a system (nGPM) for the co-delivery of GrB and PFN. Therefore, GrB and PFN were loaded in a porous polymeric nanocapsule rich in acetylcholine analogues and matrix metalloproteinase-2 (MMP-2) responsive peptides. The neutrally charged nGPM nanocapsules showed as long circulating time and accumulated at the tumour sites. Once in the tumour the outside shell of nanocapsules became degraded by overexpressed MMP-2 proteases, resulting in the release of GrB and PFN. We found that the PFN complex formed small pores on the surface of tumour cells which allow GrB to enter the cytoplasm of tumour cells inducing cell apoptosis and tumour suppression significantly.

Association between high galectin expression and poor prognosis in hematologic cancers: a systematic review and meta-analysis.

BACKGROUND: Galectin (Gal) is considered a promising immune checkpoint molecule. More and more studies have shown that high expression levels of galectins in hematologic cancer are positively correlated with poor clinical prognosis. However, the exact prognostic significance of galectins remains unclear. METHODS: PubMed, Embase, Web of Science, and Cochrane Library were searched for studies addressing the correlation of galectin expression levels with prognosis of hematologic cancers. Stata software was used to estimate hazard ratios (HR) and 95% confidence intervals (CI). RESULT: Hematologic cancer patients with high galectin expression levels showed poor overall survival (OS, HR = 2.43, 95% CI: 1.95, 3.04), disease-free survival (DFS, HR = 3.29, 95% CI: 1.61, 6.71), and event-free survival (EFS, HR = 2.20, 95% CI: 1.47, 3.29) outcomes. Subgroup analysis revealed that high expression levels of galectins pointed to relatively poor OS in MDS (HR = 5.44, 95% CI: 2.09, 14.18), as compared to AML, CHL and CLL. No correlation was found between galectins and OS in NHL and MM. Among the three galectins, Gal-9 (HR = 3.60, 95% CI: 2.03, 6.38) showed higher correlation with poor prognosis than Gal-1 and Gal-3. In addition, use of peripheral blood (HR = 2.96, 95% CI: 2.07, 4.22) samples and qRT-PCR (HR = 2.80, 95% CI: 1.96, 4.01) method for galectin detection were shown to improve its prognostic correlation in hematologic cancers. CONCLUSION: Meta-analysis revealed high expression of galectins was associated with poor prognosis in hematologic cancer patients and galectins can be considered a promising prognostic predictive marker.

Prognostic role of serum soluble ST2 of advanced breast cancer patients: a retrospective cohort study.

Background: The soluble suppression of tumorigenicity 2 receptor (sST2) binds to interleukin-33 (IL-33) and blocks IL-33 and ST2 binding, suggesting that sST2 acts as a "decoy" receptor for IL-33 and is involved in the malignant progression of breast cancer. This paper aimed to investigate the differences in sST2 expression in patients with different molecular subtypes of breast cancer and assess its clinical value in the prognostic evaluation of advanced breast cancer. Methods: In this paper, we collected sera from 91 patients firstly diagnosed with advanced breast cancer at the Tianjin Medical University Cancer Institute and Hospital from 2008 to 2021 during their first hospitalization. We detected the expression level of sST2 in the serum of patients with different molecular fractions of breast cancer and analyzed the relationship between serum sST2 levels and breast cancer-related bone metastasis, cardiotoxicity, and overall survival. Bone metastases were detected using the Emission Computed Tomography technique, and chemotherapy drug-induced cardiotoxicity was detected by echocardiography. The Overall Survival was evaluated using the Kaplan-Meier estimator, and multivariate Cox regression analysis was performed to demonstrate the association between variables and sST2 levels. Results: Serum sST2 levels did not vary among the different pathological types, molecular types, and estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor 2 (HER2), and tumor proliferation marker (Ki-67) subgroups, and only differed significantly in the cardiotoxicity group. There were no statistical differences in tissue polypeptide specific antigen (TPSA), carbohydrate antigen 125 (CA125), carcinoembryonic antigen (CEA), carbohydrate antigen 153 (CA153), D-dimer, and inflammatory indexes neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR) among different molecular subgroups. However, except for triple-negative breast cancer (TNBC), there was no difference in peripheral blood soluble ST2 concentrations among other molecular subtypes. Meanwhile, the survival of the high sST2 level group among advanced breast cancer patients was significantly lower than that of the normal group, and sST2 expression was closely related to cardiotoxicity and clinical stage. Conclusions: Serum sST2 is not only traditionally applied to the assessment of cardiac injury, but can also be utilized for assessing the prognosis of advanced breast cancer, excluding the influence of clinical stage and cardiotoxicity.

Establishment of a homogeneous immunoassay-light-initiated chemiluminescence assay for detecting anti-Müllerian hormone in human serum.

Anti-Müllerian hormone (AMH) is known as a reliable marker of ovarian reserve (OR). The determination of AMH is of great importance and most existed AMH detection methods are heterogeneous immunoassay. In this study, a novel homogeneous sandwich immunoassay-light-initiated chemiluminescence assay (LICA) for detecting AMH serum level was developed. This AMH-LICA was performed by incubating serum samples with AMH mouse monoclonal antibody coated with chemibeads, streptavidin-coated sensibeads, and biotinylated AMH mouse monoclonal antibody. Sensitivity, precision, accuracy and cross-reactivity of this assay were evaluated. Besides, a regression analysis showed a high correlation between AMH-LICA and Roche Elecsys® AMH assay (y = 0.9851x + 0.07147, R2 = 0.9569). As a homogeneous immunoassay, this AMH-LICA could accurately and rapidly determine the serum level of AMH with high-throughput. Thus, this new developed assay may be a new useful analytical tool for the determination of AMH.

Study on the Photocatalysis Mechanism of the Z-Scheme Cobalt Oxide Nanocubes/Carbon Nitride Nanosheets Heterojunction Photocatalyst with High Photocatalytic Performances.

An efficient Z-scheme Co3O4/g-C3N4 heterojunction photocatalyst was developed via in-situ forming Co3O4 nanocubes on the g-C3N4 nanosheet in the hydrothermal process. The obtained photocatalyst exhibited high photocatalytic activity for the visible-light-driven catalytic reduction of Cr(VI) and catalytic oxidation of tetracycline (TC). Among the as-synthesized catalysts, Co3O4/g-C3N4-0.04 (the mass ratio of g-C3N4 to Co3O4 is 0.04) sample exhibits the most efficient catalytic activities. The photocatalytic reduction and photocatalytic oxidation efficiencies of Co3O4/g-C3N4-0.04 can obtain 81.3 and 92.6 %, respectively. Moreover, the TC is mineralized in the course of photocatalytic degradation, 72.2% of TOC is removed from the reaction system. In addition, the apparent quantum efficiency for the removal of Cr(VI) was also obtained and the the Co3O4/g-C3N4-0.04 could achieve the highest apparent quantum efficiency among the samples. The enhancing photocatalytic activities originated from the efficient interfacial charge migration and separation obtained in Co3O4/g-C3N4-0.04, which is preliminarily confirmed by the photoluminescence spectra, time-resolved photoluminescence spectra and the photoelectrochemical characterizations. Finally, we speculate that the Co3O4/g-C3N4 heterostructures follow a more reasonable Z-scheme charge transfer in this study, which is confirmed by analyzing the results of electron paramagnetic resonance, radical scavenging experiments, and theoretical calculations.

A light-initiated chemiluminescent assay for rapid quantitation of allergen-specific IgG4 in clinical samples.

BACKGROUND: An increase in allergen-specific IgG4 (sIgG4), which serves as a blocking antibody, is associated with acquisition of immune tolerance after immunotherapy. In this study, we developed a rapid, sensitive, and homogeneous immunoassay based on the light-initiated chemiluminescent assay (LICA) technology for quantifying allergen sIgG4 in serum samples. METHODS: Allergen sIgG4 was measured in vitro by incubating the sample with biotinylated allergens and chemiluminescent beads coated with anti-human IgG4 antibody, followed by the addition of streptavidin-coated sensitizer beads. Multiple tests were performed to optimize the working conditions of the LICA and evaluate its performance. RESULTS: We established the optimal concentration of biotinylated allergens (250 ng/mL), the optimal dilution range (1:8 for Gal d 1, Gal d 2 sIgG4 and 1:4 for Gal d 3, Gal d 4 sIgG4), and the optimal incubation time (20 min for Gal d 1, Gal d 2 sIgG4 and 40 min for Gal d 3, Gal d 4 sIgG4). The lower limit of quantification (LLOQ) was 0.261 ng/mL. The coefficient variation (CV) of the LICA was <10%. The assay was unaffected by general interfering substances at physiological concentrations. It exhibited excellent accuracy to detect allergen-sIgG4 in human serum. Additionally, we demonstrated that the levels of Gal d 1, Gal d 2, and Gal d 3-sIgG4 were significantly higher in the egg allergy group (p < .05), but no differences were found between the groups for Gal d 4-sIgG4. CONCLUSIONS: The LICA demonstrated satisfactory performance and can be used for quantifying allergen sIgG4 in clinical practice.

Ultrafine Ag Nanoparticles Encapsulated by Covalent Triazine Framework Nanosheets for CO2 Conversion.

This paper describes the fabrication of covalent triazine framework nanosheet-encapsulated Ag nanoparticles (Ag0@CTFN) via a simple combination of the ultrasonic exfoliation and solution infiltration method. The as-prepared Ag0@CTFN displays an order layered-sheet structure with abundant micropores and mesopores, whereas ultrafine Ag nanoparticles are confined and stabilized in their interlayers through the interaction between N sites of triazine units and Ag nanoparticles. Considering that the Ag0@CTFN possesses the merits of high nitrogen, low density, and abundant basic sites, it was thus believed to have enough abilities to adsorb and activate CO2 in the CO2 conversion and catalysis. Importantly, the Ag0@CTFN, as a heterogeneous catalyst, showed highly catalytic activity in the carboxylation of various alkynes with CO2 at ambient pressure and low temperature. This catalyst also exhibited good functional group tolerance and excellent stability without any significant loss of its activity after six recycles. This work not only achieves valuable and novel composite material but also provides the first application of covalent triazine framework nanosheets in chemical conversion of CO2, opening a new field in preparing recyclable heterogeneous catalysts to accelerate the utilization of CO2.

Component resolved diagnostic study of cow's milk allergy in infants and young children in northern China.

BACKGROUND: Increasing dairy consumption in China has been accompanied by rising incidence of milk allergy. Here we analyzed profiles of specific immunoglobulin E (sIgE) against cow's milk proteins, and assessed their value for milk allergy diagnosis among infants and young children from northern China. METHODS: Sera collected from 48 patients with milk allergy and 27 negative control subjects was analyzed by enzyme-linked immunosorbent assay to measure sIgE to α-lactalbumin (Bos d 4), ß-lactoglobulin (Bos d 5), α-casein (Bos d 9), ß-casein (Bos d 11), and κ-casein (Bos d 12). RESULTS: Among milk-allergic individuals, most were sensitized to at least one milk protein; about half were sensitized to Bos d 5, Bos d 9, Bos d 11 and Bos d 12, respectively, while few had positive serum sIgE against Bos d 4. Bos d 12 sIgE had the largest area under curve (AUC) (0.878; 95% CI, 0.800-0.957) and thus showed the best diagnostic performance in discriminating between milk-allergic and non-milk allergic patients, with a sensitivity of 92.6% and specificity of 72.9% using a statistically optimal cut-off value (OD450nm, 0.191). The combinations of Bos d 5 + Bos d 12 showed an AUC of 0.926, which was larger than for any individual components. CONCLUSIONS: Our results revealed inter-individual variation in the sensitization to different milk allergen component. Bos d 12 sIgE showed best performance in diagnosing milk allergy. Milk allergy diagnostic accuracy was further improved using combinations of milk allergen components by application of ROC curves based on logistic regression.

Identification of the major allergenic epitopes of Eriocheir sinensis roe hemocyanin: A novel tool for food allergy diagnoses.

Crab meat and roe are highly nutritious delicacies in China. While extensive research has been conducted for allergens derived from crab-meat, data relevant to the allergenic potential of crab roe derived proteins, of which hemocyanin is a principal contender, are almost entirely absent. Using bioinformatics prediction and IgE-binding assays, the three principal immunodominant epitopes of hemocyanin were identified and then combined as a single recombinant fusion protein (rHc). This together with the full-length recombinant protein (Hc) were expressed in Escherichia coli and subsequently identified by SDS-PAGE and immunoblotting. Ninety-five percent of our patients were found to carry rHc-specific IgE antibodies by ELISA. Dot-blot inhibition, together with ELISA inhibition studies, showed that pre-incubation of patient sera with the recombinant epitope protein could inhibit26% to 63% (mean: 50%) of IgE binding to immobilized, full-length Hc and the dose-response curve represents as a sigmoid shape. The recombinant protein (rHc) represents a versatile biologic tool with which to diagnose and investigate therapies for E. sinensis allergy.

Identification and characterization of ovary development-related protein EJO1 (Eri s 2) from the ovary of Eriocheir sinensis as a new food allergen.

SCOPE: Crab is a major source of shellfish allergens. Most studies have focused on allergens in crab muscle (CM) rather than on allergens in crab ovary (CO). This study aimed to identify potential allergens in CO from Eriocheir sinensis. METHODS AND RESULTS: Dot blot and immunoblotting results revealed the differential reactivity of CM and CO extracts to positive sera from patients allergic to crabs. Four CO proteins showed good specific IgE-binding activities in 2-DE/immunoblotting analysis; mass spectrometry identified the proteins as hemocyanin, vitellogenin, ovary development-related protein EJO1and EJO2. The recently identified allergen EJO1 is named 'Eri s 2' in the World Health Organization and International Union of Immunological Societies (WHO/IUIS) allergen nomenclature database. Recombinant Eri s 2 exhibited good reactivity to positive sera, and pre-incubation with recombinant Eri s 2 abrogated the reactivity of positive sera from two patients to CO in a dose-dependent manner. Moreover, co-incubation of recombinant Eri s 2 with patient basophils dose-dependently promoted basophil activation, confirming the biological activity of Eri s 2. CONCLUSION: CO tissue is an important allergen source, and Eri s 2 is a new crab allergen. This study provides insights that will be useful for component-resolved diagnostics for crab allergy.

IRE1α-XBP1 signaling pathway, a potential therapeutic target in multiple myeloma.

Multiple myeloma (MM), which arises from the uncontrolled proliferation of malignant plasma cells, is the second most commonly diagnosed hematologic malignancy in the United States. Despite the development and application of novel drugs and autologous stem cell transplantation (ASCT), MM remains an incurable disease and patients become more prone to MM relapse and drug resistance. It is extremely urgent to find novel targeted therapy for MM. To date, the classic signaling pathways underlying MM have included the RAS/RAF/MEK/ERK pathway, the JAK-STAT3 pathway, the PI3K/Akt pathway and the NF-KB pathway. The IRE1α-XBP1 signaling pathway is currently emerging as an important pathway involved in the development of MM. Moreover, it is closely associated with the effect of MM treatment and its prognosis. All these findings indicate that the IRE1α-XBP1 pathway can be a potential treatment target. Herein, we investigate the relationship between the IRE1α-XBP1 pathway and MM and discuss the functions of IRE1α-XBP1-targeted drugs in the treatment of MM.

Sarsaparilla (Smilax Glabra Rhizome) extract inhibits migration and invasion of cancer cells by suppressing TGF-ß1 pathway.

Sarsaparilla, also known as Smilax Glabra Rhizome (SGR), was shown to modulate immunity, protect against liver injury, lower blood glucose and suppress cancer. However, its effects on cancer cell adhesion, migration and invasion were unclear. In the present study, we found that the supernatant of water-soluble extract from SGR (SW) could promote adhesion, inhibit migration and invasion of HepG2, MDA-MB-231 and T24 cells in vitro, as well as suppress metastasis of MDA-MB-231 cells in vivo. Results of F-actin and vinculin dual staining showed the enhanced focal adhesion in SW-treated cells. Microarray analysis indicated a repression of TGF-ß1 signaling by SW treatment, which was verified by real-time RT-PCR of TGF-ß1-related genes and immunoblotting of TGFBR1 protein. SW was also shown to antagonize TGF-ß1-promoted cell migration. Collectively, our study revealed a new antitumor function of Sarsaparilla in counteracting invasiveness of a subset of cancer cells by inhibiting TGF-ß1 signaling.

Daucosterol inhibits cancer cell proliferation by inducing autophagy through reactive oxygen species-dependent manner.

AIMS: This study aims to evaluate the anti-cancer effect of daucosterol and explore its possible mechanism. MAIN METHODS: MTT and colony formation assay were performed to determine the effect of daucosterol on cancer cell proliferation in vitro. H22 allograft model was used for the assessment of its anti-cancer activity in vivo. Intracellular generation of reactive oxygen species (ROS) was measured using DCFH-DA probe with flow cytometry system and a laser scanning confocal microscope. LC3 (microtubule-associated protein 1 light chain 3)-II conversion was monitored with immunofluorescence and immunoblotting to demonstrate daucosterol-induced autophagy. KEY FINDINGS: We found that daucosterol inhibits the proliferation of human breast cancer cell line MCF-7 and gastric cancer cell lines MGC803, BGC823 and AGS in a dose-dependent manner. Furthermore, daucosterol inhibits murine hepatoma H22 cell growth in ICR mice. Daucosterol treatment induces intracellular ROS generation and autophagy, but not apoptotic cell death. Treatment with ROS scavenger GSH (reduced glutathione), NAC (N-acetyl-l-cysteine) or autophagy inhibitor 3-Methyladenine (3-MA) counteracted daucosterol-induced autophagy and growth inhibition in BGC823 and MCF-7 cancer cells. SIGNIFICANCE: Daucosterol inhibits cancer cell proliferation by inducing autophagy through ROS-dependent manner and could be potentially developed as an anti-cancer agent.

Sarsaparilla (Smilax Glabra Rhizome) Extract Inhibits Cancer Cell Growth by S Phase Arrest, Apoptosis, and Autophagy via Redox-Dependent ERK1/2 Pathway.

Cancer is still the major cause of death across the world. Regular approaches cannot effectively solve the emerging problems, including drug/radiation resistance, side effects, and therapeutic ineffectiveness. Natural dietary supplements have shown effectiveness in the prevention and treatment of cancer. Sarsaparilla (Smilax Glabra Rhizome) has growth-inhibitory effects on several cancer cell lines in vitro and in vivo, with little toxicity on normal cells. However, the mechanism underlying its function remains elusive. In the present study, we examined the anticancer activity of the supernatant of the water-soluble extract (SW) from sarsaparilla. Liquid chromatography/mass spectrometry-ion trap-time-of-flight (LC/MS-IT-TOF) analysis identified flavonoids, alkaloids, and phenylpropanoids as the major bioactive components of SW. SW was shown to markedly inhibit the growth of a broad spectrum of cancer cell lines in the in vitro and in vivo assays. S phase arrest, autophagy, or/and apoptosis were partly responsible for SW-induced growth inhibition. Results of microarray analysis and validation by quantitative RT-PCR indicated the involvement of oxidative stress and the MAPK1 pathway in SW-treated cells. We further found that SW destroyed intracellular-reduced glutathione/oxidized glutathione (GSH/GSSG) balance, and supplement with N-acetylcysteine (NAC) or glutathione (GSH) significantly antagonized SW-induced S phase arrest, apoptosis, and autophagy. In addition, SW-induced GSH/GSSG imbalance activated the ERK1/2 pathway, which contributed to SW-induced S phase arrest, apoptosis, autophagy, and resultant growth-inhibitory effect. Together, our results provide a molecular basis for sarsaparilla as an anticancer agent.