Asymmetric-backed multi-frequency ultrasonic transducer for conformal tumor ablation.

OBJECTIVE: Minimally invasive ultrasound ablation transducers have been widely studied. However, conventional designs are limited by the single working frequency, restricting their conformal ablation ability (i.e. ablation size and shape controllability). METHODS: New multi-frequency ultrasonic transducer design method is proposed based on the asymmetric backing layer, which divides the transducer into non-backing-layer region (i.e. front-piezoelectric region) and backing-layer region (i.e. front-piezoelectric-backing region) with multiple local thickness mode resonant frequencies. Ablation zone can be controlled by exciting the local resonance within or between the regions, and its control flexibility is further enhanced by driven under a multi-frequency modulation signal. Experiments and calculations are combined for verifying the proposal. RESULTS: The fabricated transducer with a Y-direction asymmetric backing layer shows five resonances, with two in each region and one resonance excited in both regions. Spatial ultrasound emission is demonstrated by acoustic measurements. Tissue ablation experiments verified spatial ablation zone control, and frequency modulation driving method enables the spatial transition of ablation zone from one region to the other, generating different ablation sizes and shapes. Finally, patient-specific simulations verified the effectiveness of conformal ablation. CONCLUSION: The proposed transducer enables flexible control of ablation zone. SIGNIFICANCE: This study demonstrates a new method for conformal tumor ablation.

Preparation, characterization and cytotoxicity assessment of a novel selenized polysaccharide from Morchella sextelata.

Selenylation modification has been widely developed to improve the biological effects of natural polysaccharides. In this study, a purified new polysaccharide (MSP-4) was isolated from Morchella Sextelata, and selenized into SeMSP-4 using the HNO3-Na2SeO3 method. The selenium (Se) content of SeMSP-4 was 101.81 ± 9.90 mg/kg, and the molecular weight of SeMSP-4 was 1.23 × 105 Da. The FT-IR, XRD and AFM results showed that MSP-4 was successfully combined with the Se element. The structure characters of SeMSP-4 were analyzed by methylation analysis combined with 1D and 2D NMR spectroscopy. And, the radical scavenging test revealed that SeMSP-4 exhibited higher antioxidant capacities in vitro than MSP-4. The cytotoxicity analysis indicated that SeMSP-4 could dose-dependently inhibit the proliferation of HepG2 and HeLa cells, but did not show a cytotoxic effect on normal cells (HEK293). Furthermore, SeMSP-4 stimulation significantly increased the macrophage viability and enhanced NO production in macrophage cells. This study suggested that SeMSP-4 could be utilized as a potential selenium source with antioxidant, antitumor, and immunostimulatory activities.

A novel CPT1A covalent inhibitor modulates fatty acid oxidation and CPT1A-VDAC1 axis with therapeutic potential for colorectal cancer.

Colorectal cancer (CRC) is a common and deadly disease of the digestive system, but its targeted therapy is hampered by the lack of reliable and specific biomarkers. Hence, discovering new therapeutic targets and agents for CRC is an urgent and challenging task. Here we report that carnitine palmitoyltransferase 1A (CPT1A), a mitochondrial enzyme that catalyzes fatty acid oxidation (FAO), is a potential target for CRC treatment. We show that CPT1A is overexpressed in CRC cells and that its inhibition by a secolignan-type compound, 2,6-dihydroxypeperomin B (DHP-B), isolated from the plant Peperomia dindygulensis, suppresses tumor cell growth and induces apoptosis. We demonstrate that DHP-B covalently binds to Cys96 of CPT1A, blocks FAO, and disrupts the mitochondrial CPT1A-VDAC1 interaction, leading to increased mitochondrial permeability and reduced oxygen consumption and energy metabolism in CRC cells. We also reveal that CPT1A expression correlates with the survival of tumor-bearing animals and that DHP-B exhibits anti-CRC activity in vitro and in vivo. Our study uncovers the molecular mechanism of DHP-B as a novel CPT1A inhibitor and provides a rationale for its preclinical development as well as a new strategy for CRC targeted therapy.

Heliangin acts as a covalent ligand of RPS2 that disrupts pre-rRNA metabolic processes in NPM1-mutated acute myeloid leukemia.

Although NPM1 mutations are frequently found in acute myeloid leukemia patients, therapeutic strategies are scarce and unsuitable for those who cannot tolerate intensive chemotherapy. Here we demonstrated that heliangin, a natural sesquiterpene lactone, exerts favorable therapeutic responses in NPM1 mutant acute myeloid leukemia cells, with no apparent toxicity to normal hematogenous cells, by inhibiting their proliferation, inducing apoptosis, causing cell cycle arrest, and promoting differentiation. In-depth studies on its mode of action using quantitative thiol reactivity platform screening and subsequent molecular biology validation showed that the ribosomal protein S2 (RPS2) is the main target of heliangin in treating NPM1 mutant AML. Upon covalent binding to the C222 site of RPS2, the electrophilic moieties of heliangin disrupt pre-rRNA metabolic processes, leading to nucleolar stress, which in turn regulates the ribosomal proteins-MDM2-p53 pathway and stabilizes p53. Clinical data shows that the pre-rRNA metabolic pathway is dysregulated in acute myeloid leukemia patients with the NPM1 mutation, leading to a poor prognosis. We found that RPS2 plays a critical role in regulating this pathway and may be a novel treatment target. Our findings suggest a novel treatment strategy and lead compound for acute myeloid leukemia patients, especially those with NPM1 mutations.

Covalent targeting the LAS1-NOL9 axis for selective treatment in NPM1 mutant acute myeloid leukemia.

Patients with NPM1 gene mutation-associated acute myeloid leukemia (AML), particularly those over the age of 60, have no viable targeted therapeutic choices. In this study, we identified HEN-463, a sesquiterpene lactone derivative specific targets AML with this gene mutation. This compound inhibits the interaction of LAS1-NOL9 by covalently binding to the C264 site of the ribosomal biogenesis-related protein LAS1, which translocates the LAS1 to the cytoplasm, thereby inhibiting the maturation of 28 S rRNA. This has a profound effect on the NPM1-MDM2-p53 pathway and ultimately results in the stabilization of p53. Combining this treatment with the XPO1 inhibitor Selinexor (Sel) can ideally preserve the stabilized p53 in the nucleus, considerably enhancing the efficacy of HEN-463 and addressing Sel's drug resistance. Patients with AML over the age of 60 who possess the NPM1 mutation have an unusually elevated level of LAS1, which has a significant impact on their prognosis. In NPM1-mutant AML cells, decreased LAS1 expression promotes proliferation inhibition, apoptosis, cell differentiation, and cell cycle arrest. This suggests that it may be a therapeutic target for this kind of blood cancer, especially in patients over the age of 60.

Does Methionine Status Influence the Outcome of Selenomethinione Supplementation? A Comparative Study of Metabolic and Selenium Levels in HepG2 Cells.

Methionine restriction and selenium supplementation are recommended because of their health benefits. As a major nutrient form in selenium supplementation, selenomethionine shares a similar biological process to its analog methionine. However, the outcome of selenomethionine supplementation under different methionine statuses and the interplay between these two nutrients remain unclear. Therefore, this study explored the metabolic effects and selenium utilization in HepG2 cells supplemented with selenomethionine under deprived, adequate, and abundant methionine supply conditions by using nuclear magnetic resonance-based metabolomic and molecular biological approaches. Results revealed that selenomethionine promoted the proliferation of HepG2 cells, the transcription of selenoproteins, and the production of most amino acids while decreasing the levels of creatine, aspartate, and nucleoside diphosphate sugar regardless of methionine supply. Selenomethionine substantially disturbed the tricarboxylic acid cycle and choline metabolism in cells under a methionine shortage. With increasing methionine supply, the metabolic disturbance was alleviated, except for changes in lactate, glycine, citrate, and hypoxanthine. The markable selenium accumulation and choline decrease in the cells under methionine shortage imply the potential risk of selenomethionine supplementation. This work revealed the biological effects of selenomethionine under different methionine supply conditions. This study may serve as a guide for controlling methionine and selenomethionine levels in dietary intake.

Selenium Nanoparticles Synergistically Stabilized by Starch Microgel and EGCG: Synthesis, Characterization, and Bioactivity.

Selenium (Se) is a chemical element essential to human health because of its bioactive properties, including antioxidative, anticancer, and immunomodulating activities. Despite the high therapeutic potential of Se, its intrinsic properties of poor stability, a narrow therapeutic window, and low bioavailability and bioactivity have limited its clinical applications. Selenium nanoparticles (SeNPs) exhibit lower toxicity and higher bioactivity than other Se forms. Herein, we report a green method for the preparation of monodisperse SeNPs with starch microgel (SM) and epigallocatechin gallate (EGCG) through Se-O bonds and polysaccharide-polyphenol interactions (namely, SM-EGCG-SeNPs). SM-EGCG-SeNPs showed higher stability, bioactivities, and cytotoxicity than SeNPs and SM-SeNPs at the equivalent dose. SM-EGCG-SeNPs induced the apoptosis of cancer cells via the activation of several caspases and reactive oxygen species overproduction. This work proposes a facile method for the design and potentiation of structure-bioactive SeNPs via polysaccharide-polyphenol interactions.

MiR-218-5p Suppresses the Killing Effect of Natural Killer Cell to Lung Adenocarcinoma by Targeting SHMT1.

PURPOSE: Lung adenocarcinoma (LA) is one of the major types of lung cancer. MicroRNAs (miRNAs) play an essential role in regulating responses of natural killer (NK) cells to cancer malignancy. However, the mechanism of miR-218-5p involved in the killing effect of NK cells to LA cells remains poorly understood. MATERIALS AND METHODS: The expression of miR-218-5p was examined by quantitative real-time polymerase chain reaction (qRT-PCR). Serine hydroxymethyl transferase 1 (SHMT1) level was detected by qRT-PCR or western blots. Cytokines production of interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α) were detected by ELISA. The killing effect of NK cells to LA cells was investigated using lactate dehydrogenase cytotoxicity assay kit. The interaction of miR-218-5p and SHMT1 was probed by luciferase activity assay. Xenograft model was established to investigate the killing effect of NK cells in vivo. RESULTS: miR-218-5p was enhanced and SHMT1 was inhibited in NK cells of LA patients, whereas stimulation of interleukin-2 (IL-2) reversed their abundances. Addition of miR-218-5p reduced IL-2-induced cytokines expression and cytotoxicity in NK-92 against LA cells. Moreover, SHMT1 was negatively regulated by miR-218-5p and attenuated miR-218-5p-mediated effect on cytotoxicity, IFN-γ and TNF-α secretion in IL-2-activated NK cells. In addition, miR-218-5p exhaustion inhibited tumor growth by promoting killing effect of NK cells. CONCLUSION: miR-218-5p suppresses the killing effect of NK cells to LA cells by targeting SHMT1, providing a potential target for LA treatment by ameliorating NK cells function.

Tailor-made PL-UC-C3 nanoparticles for fluorescence/computed tomography imaging-guided cascade amplified photothermal therapy.

BACKGROUND: Development of the burgeoning number of photothermal therapy (PTT) agents has drawn a huge amount of interest, since PTT treatment is a powerful and effective alternative to traditional treatments. Optimal PTT agents should integrate some essential preconditions including negligible systemic toxicity, deep penetration into tumor tissues, and maximum laser energy absorbance. Unfortunately, only few of the PTT agents reported could meet all of the above mentioned conditions. METHODS: Here, we report a brand new PTT agent through the encapsulation of NaGdF4:Yb,Tm@ NaGdF4:Yb (UCNPs) and an organic compound (C3) into poly-e-caprolactone-polyethylene-polyglycol (PCL-PEG) (PL-UC-C3 NPs). RESULTS: UCNPs as an up-conversion material and C3 as a PTT agent both feature low cytotoxicity, and most importantly, UCNPs with superior conversion efficiency could efficiently absorb the energy of a 980 nm laser, transform the near-infrared laser light into visible light, and translate the palingenetic visible light to C3. The usage of a 980 nm laser ensures a deeper penetration and lower energy, while the highly efficient absorption and transformation process confers a cascade amplified hyperthermia for tumor treatment. CONCLUSION: In this regard, our research provides a powerful and robust breakthrough for florescence/computed tomography imaging-guided PTT treatment, lighting up the clinical application in cancer treatment.

Doxorubicin and paclitaxel carried by methoxy poly(ethylene glycol)-poly(lactide-co-glycolide) is superior than traditional drug-delivery methods.

AIM: To evaluate the advantages of nanomaterial methoxy poly(ethylene glycol)-poly(lactide-co-glycolide) (mPEG-PLGA) encapsulated doxorubicin (D/DOX) and paclitaxel (T/TAX; mPEG-PLGA-DT) over free form of DOX and TAX (DOX/TAX). MATERIALS & METHODS: Metabonomics was conducted to characterize the systemic metabolic response of allograft breast cancer model mice to mPEG-PLGA-DT and DOX/TAX treatments. RESULTS: Breast tumor growth induced metabolic reprogram in serum and multiple organs. DOX/TAX treatment could ameliorate the elevated energy and nucleotides demands in some organs while mPEG-PLGA-DT treatment showed outstanding therapeutic outcomes in restoring the metabolic phenotypes of serum and kidney from tumor-bearing mice to the healthy state. CONCLUSION: This investigation proved the biological advantages of mPEG-PLGA-DT over DOX/TAX in molecular level through the comparison between their metabolic responses in vivo.

A targeted biocompatible organic nanoprobe for photoacoustic and near-infrared-II fluorescence imaging in living mice.

Multimodal molecular imaging probes have attracted much attention, and they possess great potential to accurately diagnose diseases due to the synergistic superiorities of multiple complementary imaging. Herein, a targeted biocompatible organic nanoplatform (IR-PEG-FA) with a strong optical absorption in the near-infrared window (NIR-I) for photoacoustic imaging (PAI) and excellent second near-infrared (NIR-II) fluorescence imaging property for NIR-II imaging is fabricated. The dual-modal nanoprobe is composed of the small organic dye molecule IR-1061, water-soluble poly(ethylene glycol) (PEG) and folic acid (FA) as the targeted ligands. Depending on the strength of high temporal resolution and preeminent spatial resolution, the targeted biocompatible dual-mode nanoprobe for PAI and NIR-II imaging can provide more detailed date of cancers and diseases, and enables us to specifically diagnose them through quite a precise way.

Decreased glutathione biosynthesis contributes to EGFR T790M-driven erlotinib resistance in non-small cell lung cancer.

Epidermal growth factor receptor (EGFR) inhibitors such as erlotinib are novel effective agents in the treatment of EGFR-driven lung cancer, but their clinical impact is often impaired by acquired drug resistance through the secondary T790M EGFR mutation. To overcome this problem, we analysed the metabonomic differences between two independent pairs of erlotinib-sensitive/resistant cells and discovered that glutathione (GSH) levels were significantly reduced in T790M EGFR cells. We also found that increasing GSH levels in erlotinib-resistant cells re-sensitised them, whereas reducing GSH levels in erlotinib-sensitive cells made them resistant. Decreased transcription of the GSH-synthesising enzymes (GCLC and GSS) due to the inhibition of NRF2 was responsible for low GSH levels in resistant cells that was directly linked to the T790M mutation. T790M EGFR clinical samples also showed decreased expression of these key enzymes; increasing intra-tumoural GSH levels with a small-molecule GST inhibitor re-sensitised resistant tumours to erlotinib in mice. Thus, we identified a new resistance pathway controlled by EGFR T790M and a therapeutic strategy to tackle this problem in the clinic.

Germanicol induces selective growth inhibitory effects in human colon HCT-116 and HT29 cancer cells through induction of apoptosis, cell cycle arrest and inhibition of cell migration.

PURPOSE: The main aim of this research was to evaluate the anticancer and apoptotic effects of germanicol - a natural triterpene - in HCT-116 and HT29 human colon cancer cells and deciphering its mode of action by studying its effect on the cell cycle and cell migration. METHODS: Cell cytotoxicity was evaluated by MTT assay, while cell death was assessed by LDH assay. Fluorescence microscopy, using DAPI and acridine orange/ethidium bromide (AO-ETBR), was carried out to evaluate the effect of germanicol on cellular morphology and apoptosis induction. Apoptosis quantification was performed by Annexin V-FITC assay, while cell cycle analysis was performed by flow cytometry using propidium iodide (PI). RESULTS: The results revealed that germanicol showed selective, potent and dose-dependent cytotoxicity in HCT-116 and HT29 human colon cancer cells, while it showed lower cytotoxicity in normal colon cells (human colon fibroblast, CCD-18Co). LDH assay also showed that germanicol induced dose-dependent cell death in HCT-116 and HT29 cells. Fluorescence microscopy revealed that germanicol induced apoptosis via chromatin condensation and DNA damage in HCT-116 colon cancer cells. It also revealed that the percentage of cells with orange and red fluorescence increased when adding a germanicol dose, indicating apoptosis. Germanicol also inhibited cancer cell migration. CONCLUSION: The current findings reveal that germanicol exhibits selective antiproliferative activity against two human colon cancer cells. The normal cell line was less affected by the drug, as compared to the two cancer cell lines, indicating that germanicol will not target normal living cells. The antiproliferative effect was shown to be mediated through the induction of apoptosis and suppression of cell migration.

Tumor growth affects the metabonomic phenotypes of multiple mouse non-involved organs in an A549 lung cancer xenograft model.

The effects of tumorigenesis and tumor growth on the non-involved organs remain poorly understood although many research efforts have already been made for understanding the metabolic phenotypes of various tumors. To better the situation, we systematically analyzed the metabolic phenotypes of multiple non-involved mouse organ tissues (heart, liver, spleen, lung and kidney) in an A549 lung cancer xenograft model at two different tumor-growth stages using the NMR-based metabonomics approaches. We found that tumor growth caused significant metabonomic changes in multiple non-involved organ tissues involving numerous metabolic pathways, including glycolysis, TCA cycle and metabolisms of amino acids, fatty acids, choline and nucleic acids. Amongst these, the common effects are enhanced glycolysis and nucleoside/nucleotide metabolisms. These findings provided essential biochemistry information about the effects of tumor growth on the non-involved organs.

miR-515-5p controls cancer cell migration through MARK4 regulation.

Here, we show that miR-515-5p inhibits cancer cell migration and metastasis. RNA-seq analyses of both oestrogen receptor receptor-positive and receptor-negative breast cancer cells overexpressing miR-515-5p reveal down-regulation of NRAS, FZD4, CDC42BPA, PIK3C2B and MARK4 mRNAs. We demonstrate that miR-515-5p inhibits MARK4 directly 3' UTR interaction and that MARK4 knock-down mimics the effect of miR-515-5p on breast and lung cancer cell migration. MARK4 overexpression rescues the inhibitory effects of miR-515-5p, suggesting miR-515-5p mediates this process through MARK4 down-regulation. Furthermore, miR-515-5p expression is reduced in metastases compared to primary tumours derived from both in vivo xenografts and samples from patients with breast cancer. Conversely, miR-515-5p overexpression prevents tumour cell dissemination in a mouse metastatic model. Moreover, high miR-515-5p and low MARK4 expression correlate with increased breast and lung cancer patients' survival, respectively. Taken together, these data demonstrate the importance of miR-515-5p/MARK4 regulation in cell migration and metastasis across two common cancers.

Assessment of the Biological Effects of a Multifunctional Nano-Drug-Carrier and Its Encapsulated Drugs.

Polymer-nanoparticle-encapsulated doxorubicin (DOX) and paclitaxel (TAX) have the potential for novel therapeutic use against cancer in the clinic. However, the systemic biological effect of the nanoparticle material, namely, methoxypoly(ethylene glycol)-poly(lactide-co-glycolide) (mPEG-PLGA), and its encapsulated drugs have not been fully studied. We have applied NMR-based metabonomics methodology to characterize and analyze the systemic metabolic changes in mice after being exposed to mPEG-PLGA, mPEG-PLGA-encapsulated DOX and TAX (NP-D/T), and their free forms. The study revealed that mPEG-PLGA exposure only induces temporary and slight metabolic alternations and that there are detoxification effects of nanoparticle packed with D/T drugs on the heart when comparing with free-form D/T drugs. Both NP-D/T and their free forms induce a shift in energy metabolism, stimulate antioxidation pathways, and disturb the gut microbial activity of the host. However, mPEG-PLGA packaging can relieve the energy metabolism inhibition and decrease the activation of antioxidation pathways caused by D/T exposure. These findings provide a holistic insight into the biological effect of polymer nanoparticle and nanoparticle-encapsulated drugs. This study also furthers our understanding of the molecular mechanisms involved in the amelioration effects of mPEG-PLGA packaging on the toxicity of the incorporated drugs.

Selective metabolic effects of gold nanorods on normal and cancer cells and their application in anticancer drug screening.

Cetyltrimethylammonim bromide coated gold nanorods (Au NRs) has a potential to become anti-cancer nano-drugs. Previously, the comparative responses of human alveolar adenocarcinoma epithelial cells (A549) and normal bronchial epithelial cells (16HBE) exposed to Au NRs have been characterized. It has been shown that Au NRs are translocated from the lysosome to the mitochondria in A549 cells but not in normal 16HBE cell lines. However, the molecular information during this cellular translocation remains largely undetermined. Here, we have used a metabonomic technique to comparatively analyze the time-dependent metabolic changes in Au NRs-induced A549 and 16HBE. We found that Au NRs exposure caused a disruption in the intracellular environment of both A549 and 16HBE cells, which metabolically manifested in the reduction of lactate levels in both cell lines. In addition, Au NRs induced oxidative stress in both cells lines. However, the 16HBE cells are more able to offset the oxidative stress than the A549 cells; this is because de novo GSH synthesis is triggered in Au NRs treated 16HBE cells but not in A549 cells, and the conversion of GSH to GSSG is more profound in 16HBE cells compared to A549 cells. The severe oxidative stress induces damage to mitochondria in A549 cells, leading to cell death, which is evident in the marked reduction in the levels of nucleosides and nucleotides. Furthermore, significantly elevated levels of amino acids are likely due to stress hormones being produced in Au NRs treated cells. These findings provide comprehensive molecular information on the distinctive intracellular localization, cellular uptake and translocation of Au NRs in normal and tumor cells, highlighting the value of metabonomics in assessing biological effects of nano-drugs.

Simultaneous operative treatment of patients with primary liver cancer associated with portal hypertension.

OBJECTIVE: To explore the operative procedure for patients with primary liver cancer associated with portal hyper tension (PLCPH). METHODS: We analyzed retrospectively the effect of operative procedure for 9 patients with PLCPH complicated by severe esophageal varicosity and hypersplenism. RESULTS: All patients underwent liver resection and pericardiac devascularization with splenectomy. Of the 9 patients, 2 died from liver cancer recurrence separately 13 and 16 months after operation, and 1 died from massive duodenal ulcer bleeding and multiple organs failure. Six patients survived 3, 4, 8, 10, 12 and 25 months after operation. CONCLUSIONS: The patients with PLCPH undergoing simultaneous operation could acquire curative effect as compared with those who underwent liver resection. This operation is beneficial to the patients with poor liver function.