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Recently, multifunctional nanoparticles have shown great prospects in cancer treatment, which have the ability to simultaneously deliver the drug, image and target tumor cells. In this paper, we designed a luminescent nanoparticles platform based on hydrothermal hyaluronic acid/amorphous calcium phosphate (HA-FCNs/ACP) with multifunctional properties for drug delivery, bio-imaging, and targeting treatment. HA-FCNs/ACP shows an ability to load curcumin (Cur) with pH-sensitive responsive drug release behavior and excellent biocompatibility. HA-FCNs/ACP dispersed in the cytoplasm through the overexpressed CD44 receptor that is actively targeted into human lung cancer cells (A549 cells). Meanwhile, the viability of A549 cells was significantly inhibited in vitro. The prepared HA-FCNs and HA-FCNs/ACP both exhibit excellent targeted bioimaging performance on cancer cells Hence, the as-prepared nanoparticles have promising applications in treating tumor disease.

Background Oxidized LDL(Ox-LDL) mediated endothelial dysfunction is involved in the pathogenesis of various cardiovascular diseases, including atherosclerosis. Azilsartan is a potent agent for the treatment of hypertension as the antagonist of the angiotensin II receptor. This study will investigate whether Azilsartan possesses a beneficial effect against endothelial cell dysfunction induced by ox-LDL and explore the underlying preliminary mechanism. Methods Ox-LDL was applied to construct an in vitro endothelial dysfunction model in human umbilical vascular endothelial cells (HUVECs). The expression of lectin-type oxidized LDL receptor 1 (LOX-1), endothelial nitric oxide synthase (eNOS), tight junction protein occludin, and transcriptional factor Kruppel-like factor 2 (KLF2) was detected using qRT-PCR and Western blot. ELISA and qRT-PCR were utilized to evaluate the production of chemokine monocyte chemotactic protein 1 (MCP-1) and chemokine (C-X-C motif) Ligand 1 Protein (CXCL1) in treated HUVECs. The generation of nitro oxide (NO) was determined using DAF-FM DA staining assay. KLF2 was silenced by transfecting the cells with specific Small interfering RNA (siRNA). FITC-dextran permeation assay was used to check the endothelial monolayer permeability of treated HUVECs. Results Firstly, the elevated expressions of LOX-1, MCP-1, and CXCL-1 induced by stimulation with ox-LDL were significantly suppressed by Azilsartan. The downregulated eNOS and reduced production of NO induced by ox-LDL were reversed by the introduction of Azilsartan. Secondly, enlarged endothelial monolayer permeability and decreased expression of occludin stimulated with ox-LDL were greatly reversed by treatment with Azilsartan but were abolished by silencing the expression of KLF2. Lastly, the inhibited expression of KLF2 induced by ox-LDL was significantly elevated by the introduction of Azilsartan. Conclusion Azilsartan might ameliorate ox-LDL-induced endothelial damage via elevating the expression of KLF2.

The aim of the study is to interpret the effects of air-impingement jet drying (AIJD) on drying kinetics, color, polyphenols, and antioxidation ability of Boletus aereus slices. Page model was most suitable for expressing and predicting AIJD curves of B. aereus slices. The moisture-effective diffusion coefficient of AIJD ranged from 7.8876 × 10-10 to 2.1426 × 10-9 m2 /s, and AIJD also showed high efficiency due to its low activation energy (45.37 kJ/mol). AIJD is better for B. aereus slices than hot air drying (HAD) in accelerating the drying rate (DR) and shortening drying time, and maintaining color. p-hydroxybenzoic acid, protocatechuic acid, and rutin were identified in B. aereus slices by ultra high-performance liquid chromatography coupled with triple quadrupole mass spectrometry (UHPLC-QqQ-MS). Total polyphenols, flavanone, phenolic acids, and antioxidant activities were significantly lower in dried B. aereus slices than those in fresh B. aereus slices. In AIJD, drying temperature had the greatest effect on the quality of B. aereus slices, and AIJD at 50 °C is the optimum drying condition for B. aereus slices. PRACTICAL APPLICATION: Boletus aereus occurs in many countries all over the world. In this paper, the effect of AIJD on color, polyphenols, and antioxidation ability in B. aereus slices and its drying kinetics were studied. AIJD is an efficient drying method for B. aereus by decreasing its drying time, increasing DR, and protecting the color of B. aereus. These findings have provided important reference basis for people to have a better understanding of AIJD method, which was used to dry B. aereus. This study also provides a new technique for drying B. aereus, which could improve dry efficiency and reduce drying cost.

The purpose was to explore the drying kinetics, the moisture effective diffusivities, color, total polyphenols, lycopene and antioxidant activities of dried tomato slices by air-impingement jet drying (AIJD). The results showed that high temperature increased the drying rate, and Modified Page model accurately predicted the AIJD characteristics of tomato slices. AIJD is better than hot air drying in shortening drying time, enhancing drying rate and decreasing the loss of total polyphenols, lycopene and antioxidant capacity of tomato slices. Tomato slices dried by AIJD also showed higher lightness and redness. Lycopene content and antioxidant activity of tomato slices dried by AIJD were increased by higher drying temperature. Based on experimental data, AIJD at 80 °C can be used in tomato drying process due to the advantages in drying efficiency and content of bioactive compounds. This study will provide helpful information for the production of high quality of dried tomato products.

Although amorphous calcium phosphate has been a mature research object in the medicine field, its application in drug carriers remains restricted due to its easy agglomeration and poor dispersion s...

Introduction Oxidized low-density lipoprotein (ox-LDL)-induced endothelial senescence is involved in the pathogenesis of atherosclerosis and many cardiovascular diseases. G-protein-coupled receptor 120 (GPR120), a type of orphan G-protein-coupled receptors (GPRs), plays a vital role in mediating anti-inflammatory and insulin-sensitizing effects. The biological function of GPR120 in vascular endothelial cells is largely unknown. Methods The human aortic endothelial cells (HAECs) were treated with ox-LDL (100 μg/mL) in the presence or absence of GW9508 (50 μM) or AH9614 (1 μM) for 24 h. The LDH assay was used to determine cell death. The dihydroethidium (DHE) staining assay was used to measure intracellular levels of reactive oxidative species (ROS), and a senescence β-galactosidase assay kit was used to determine endothelial senescence. Gene and protein expressions were measured using real-time polymerase chain reaction (PCR) and western blot analysis, respectively. Results Ox-LDL treatment decreased the expression of GPR120 by more than half in HAECs. Typically, 100 μg/mL of ox-LDL- induced 35.2% LDH release, which was reduced to 16.9% by 50 μM GW9508, the agonist of GPR120. Importantly, GW9508 relieved cytotoxicity and suppressed the ox-LDL-induced increase in the activity of senescence-associated β-galactosidase (SA-β-Gal) (from 3.3-fold to 1.6-fold of the control group) and the generation of cellular reactive oxidative species (ROS) (from 3.8-fold to 1.6-fold of the control group). Furthermore, we found that GW9508 ameliorated ox-LDL-induced endothelial cell cycle arrest at the G0/G1 phase and the expression of key senescence proteins, including p53 and plasminogen activator inhibitor-1(PAI-1). Mechanistically, we showed that GW9508 promoted ox-LDL-induced transcriptional factor NF-E2-related factor 2 (NRF2) (increase by 47.3%) translocation into the nucleus. The effect of GW9508 is dependent on its receptor GPR120, the blockage of which by its specific antagonist, AH7614, abolished the antisenescence effect of GW9508. Conclusion Collectively, this study revealed the protective effect of GPR120 activation in vascular endothelial cells, implying that GPR120 is a promising therapeutic target for the treatment of cardiovascular diseases.

In this work, a promising propellant binder using the energetic branched glycidyl azide polymer (B-GAP) as a matrix and the low-toxic dimer acid diisocyanate (DDI) as a curing agent was prepared, under the catalysis of dibutyl tin dilaurate. The curing kinetics considering the thermal diffusion effect and the reaction endpoint of B-GAP/DDI were investigated by the thermal analysis method and a newly proposed variance method, respectively. Moreover, the buildup of microstructures during curing and the tensile and dynamic mechanical performance of this binder were respectively explored. Results show that there exists an obvious induction period in the beginning of the curing reaction, and the autocatalytic model shows that thermal diffusion can effectively describe the curing process. Shore A hardness of sample stabilizes around 40.78 in the end of curing, and the reaction endpoint of B-GAP/DDI is in the time range of 156–168 h. There exist cross-linking, suspension, and free chains during the whole curing process, and the cross-linking density of the binder reaches approximately 4.0 × 10–4 mol·cm–3 when the curing completes. Hydrogen bonding (H-bond) is found to be a strong binder: 53.3% of the carbonyls participates in forming the H-bond. Furthermore, this binder has better mechanical performance and lower glass-transition temperature than the GAP/N100 binder.

Water falling film dielectric barrier discharge (WFFDBD) has been widely studied for water treatment, due to its good performance on production of active species, mass transfer of active species from gas phase into liquid phase, and therefore the degradation of pollutants. However, few studies have focused on the production characteristics of active species of WFFDBD. In this paper, the formation characteristics of hydrogen peroxide, ozone and nitrate ions in a WFFDBD reactor powered by bipolar pulsed generator were studied under different electric discharge time, peak pulse voltage, power–frequency, air flow rate, water circulation flow rate, pH value and conductivity. The results show that the concentrations of hydrogen peroxide, ozone and nitrate ions increased with the peak pulse voltage, power supply frequency and water circulation flow rate, but the inputted air flow rate presented different effect on the production of active species. Moreover, the increase in pH value and conductivity led to a decrease in the concentration of hydrogen peroxide, while nitrate ion increased with the water pH value and conductivity. The p-nitrophenol degradation result indicates that both ozone and hydrogen peroxide contribute to its degradation possibly via their reaction into hydroxyl radical.

Insect tea is consumed as a health beverage in China. The insect tea primary leaf (ITPL) is rich in bioactive substances, which are also used as traditional Chinese medicine. This study investigated the role of ITPL in reducing the oxidative response induced by D‐galactose in mice. Mice were intraperitoneally injected with D‐galactose to induce oxidative damage. The effect of ITPL was tested by pathological observation, serum detection with kits, quantitative polymerase chain reaction, and Western blot. The experimental results show that ITPL increased the thymus, brain, heart, liver, spleen, and kidney indices of oxidized mice. ITPL increased superoxide dismutase, glutathione peroxidase, and glutathione levels and reduced nitric oxide and malondialdehyde levels in the serum, liver, and spleen in oxidative damaged mice. The pathological observations show that ITPL reduced the oxidative damage of the liver and spleen in mice induced with D‐galactose. Simultaneously, ITPL upregulated mRNA expression of neuronal nitric oxide synthase, endothelial nitric oxide synthase, cuprozinc‐superoxide dismutase, manganese superoxide dismutase, catalase, heme oxygenase‐1, nuclear factor‐erythroid 2 related factor 2, γ‐glutamylcysteine synthetase, and NAD(P)H dehydrogenase [quinone] 1, and downregulated the expression of inducible nitric oxide synthase in the liver and spleen of oxidized mice. ITPL had beneficial preventive effects on the oxidative damage caused by D‐galactose in mice and was more effective as an antioxidant than vitamin C. The component analysis test by high‐performance liquid chromatography indicated that ITPL contained the following seven compounds: neochlorogenic acid, cryptochlorogenic acid, rutin, kaempferin, isochlorogenic acid B, isochlorogenic acid A, and hesperidin. ITPL is a plant with excellent antioxidant activities derived from its bioactive substances., Insect tea primary leaf (Malus sieboldii (Regal) Rehd.) extract could inhibit D‐galactose‐induced oxidative damages in mice.

Cardiovascular disease (CVD) has been considered as a major risk factor of death in recent decades. In CVDs, the NLRP3 inflammasome is important for inflammatory response and vascular damage. Therefore, safe and effective treatments to decrease NLRP3 inflammasome activation are required. Increased levels of free fatty acid (FFA) have been associated with the progression of CVD. Humanin, a kind of mitochondrial-derived peptide, has shown its beneficial effects in different types of cells. However, the roles of humanin in the NLRP3 inflammasome induced by FFA are still unknown. Here, we investigated the molecular mechanisms whereby humanin was found to exert protective effects in human aortic endothelial cells (HAECs) against FFA-caused endothelial injury. Here, treatment with humanin inhibited FFA-induced lactate dehydrogenase release, thereby demonstrating a protective capacity against cell death. Humanin also suppressed oxidative stress by downregulating the expression of reactive oxygen species and NOX2. Notably, humanin reduced NLRP3 and p10 and rescued FFA-induced dysfunction of adenosine monophosphate-activated protein kinase. Consequently, humanin inhibited the expression of IL-1β and IL-18. These results conclude that humanin might be a promising therapeutic agent for CVD.

The Malus pumila leaves are used as a kind of tea drink in China, and there are abundant flavonoids in the leaves of Malus pumila. In this study, BALB/c mice received senna extract solution via gavage, which induced acute diarrhea, and the interventive effect of Malus pumila leaf flavonoids (MPLF) was observed. The results showed that MPLF decreased the diarrhea index, and MPLF also decreased the capillary permeability in the abdominal cavity of mice. The result of serum cytokine detection indicated that MPLF decreased the levels of inflammatory cytokines, including interleukin 6 (IL‐6), interleukin 12 (IL‐12), and tumor necrosis factor alpha (TNF‐α). The results of quantitative polymerase chain reaction (qPCR) indicated that diarrhea decreased the mRNA and protein expression of aquaporin‐3 (AQP3) in the jejunum and aquaporin‐4 (AQP4) in the ileum, which were inhibited by MPLF. By high performance liquid chromatography (HPLC), MPLF mainly contains 10 flavonoids, which are rutin, hyperoside, isoquercitrin, taxifolin, quercitrin, hesperidin, myricetin, baicalin, neohesperidin dihydrochalcone and quercetin, the synergistic effect of these components plays an antidiarrhea role in MPLF. Therefore, MPLF achieved good antidiarrheal effect, which was better than that of the commonly used montmorillonite powder at the same concentration. MPLF is a natural raw material for functional food with an antidiarrheal effect., Malus pumila leaf flavonoids inhibited senna‐induced acute diarrhea in BALB/c mice through up‐regulation AOP3 mRNA and protein expression in jejunum tissue and AQP4 expression in ileum tissue.

Despite the long-term use and recent development of keratin materials, the mechanism of their solubilization is still not well understood beyond the breakage of disulfide bonds due to the complexit...

Nanozymes are inorganic nanoparticles that mimic the enzyme-like properties in redox reactions, processing both unique properties of nanomaterials and a catalytic function. Because of high catalytic activity, stability and multifunctionality, nanozyme are of increasingly wide interest in the fields of environmental science and technology. In this article, we review the most recent advances of nanozyme research for environmental pollutant detection and treatment. Nanozymes can be used to detect ions, molecules and organic compounds both qualitatively and quantitatively. They have also been applied for destruction multi-drug resistant bacteria and the degradation of various organic pollutants. Despite the apparent potential of nanozymes in environmental science and technology, current research and application is still limited, and so future challenges and research prospects have been highlighted.

This study was conducted to investigate the preventative effect of Lactobacillus fermentum HFY06 on carbon tetrachloride (CCl4)-induced liver injury in Kunming mice. Mice were treated with HFY06, then liver damage was induced using CCl4. Evaluation indicators included the activities of aspartate aminotransferase (AST), triglycerides (TG), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and malondialdehyde (MDA) in serum; cytokines levels of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ) in serum; and related gene expressions of nuclear factor-κB (NF-κB), TNF-α, cyclooxygenase-2 (COX-2), copper/zinc superoxide dismutase (Cu/Zn-SOD), manganese superoxide dismutase (Mn-SOD), and catalase (CAT). Liver tissue was stained with hematoxylin and eosin for pathological analysis. Compared with the model group, HFY06 reduced the liver index, increased the serum SOD and GSH-Px activities, and reduced the AST, TG, and MDA activities in the mice. Inflammation-related IL-6, TNF-α and IFN-γ levels were also reduced after treatment with a high dose of HFY06. Pathological observation showed that CCl4 damaged the mouse livers, which were significantly improved after treatment with silymarin and HFY06. qPCR also confirmed that the high dose of HFY06 (109 colony-forming units [CFU] per kg per day) upregulated the mRNA expression of the antioxidant genes, Cu/Zn-SOD, Mn-SOD, and CAT, in the liver tissue and downregulated the mRNA expression of the inflammatory factors, NF-κB, TNF-α and COX-2, but HFY06 was less effective than silymarin. These findings indicate that HFY06 prevented CCl4-induced liver damage in vivo but was less effective than silymarin. Thus, HFY06 may have a potential role in treating liver diseases.

The aim of this study was to compare the phenolic compound profiles and antioxidant capacities of eight varieties of longan (Dimocarpus longan Lour.) planted in the middle and upper Yangtze River area. The total polyphenols content (TPC) and total flavonoids content (TFC) of dried longan pulp ranged from 1.07 ± 0.05 to 1.22 ± 0.05 mg gallic acid equivalents/g and 0.25 ± 0.07 to 0.87 ± 0.14 mg rutin equivalents/g. UHPLC-QqQ-MS/MS analysis revealed 12 individual polyphenol compounds in longan. The Fuwan8, Dongliang and FD97 varieties showed the strongest DPPH scavenging activity (IC50 of 1.03 g/mL). The highest ABTS+ scavenging activity was found in the Dongliang. In the enzyme assays, the Fuwan8 and Dongliang varieties demonstrated maximum α-amylase and α-glucosidase inhibition activities, with values of 97.56 and 88.58%, respectively. The principal component analysis indicated that the Rongyu8 and Songfengben cultivars have nearly analogous polyphenol compounds.

The optimal synchronous conditions to extract tea polysaccharides (TPS) and tea polyphenols (TPP) from Ya'an Tibetan tea were investigated, and the antioxidative capacity of TPS and TPP was measured, and the tea was analyzed to identify the polyphenol compounds it contained. On the basis of single‐factor experiments, a Box–Behnken design and response surface methodology were applied to optimize the hot water extraction conditions. The optimal extraction technology was determined as extraction temperature of 83°C, time of 104 min, and liquid‐to‐material ratio of 41 ml/g, yielding TPP and TPS at 42.70 ± 2.38 mg/g and 53.86 ± 3.79 mg/g, respectively. The TPS and TPP in Ya'an Tibetan tea have high eliminating activities on DPPH and strong reducing power, with TPP showing a higher antioxidant activity than TPS. UHPLC‐QqQ‐MS/MS analysis identified EGCG, GCG, and ECG as major polyphenol components in Ya'an Tibetan tea. These findings might promote the application of Ya'an Tibetan tea in the food industry.

Insect tea is a traditional Chinese drink that contains abundant bioactive substances. In this study, the preventive effect of Insect tea on D‐galactose‐induced oxidation in mice was studied. The serum, liver, and spleen of mice were measured by biochemical and molecular biological methods, which showed that Insect tea could increase the biochemical indexes of the thymus, brain, heart, liver, spleen, and kidney in mice with induced oxidative damage. Insect tea can increase the levels of SOD (superoxide dismutase), GSH‐Px (glutathione peroxidase), and GSH (glutathione) and decrease the levels of MDA (malondialdehyde) in the serum, liver, and spleen of mice with oxidative damage. Pathological observation also confirmed that Insect tea could inhibit oxidative damage of the liver and spleen tissue caused by D‐galactose in mice. Further molecular biological experiments also showed that Insect tea could upregulate the mRNA and protein expression of Cu/Zn‐SOD (cuprozinc‐superoxide dismutase), Mn‐SOD (manganese superoxide dismutase), CAT (catalase), HO‐1 (heme oxygenase‐1), Nrf2 (nuclear factor‐erythroid 2 related factor 2), γ‐GCS (γ‐glutamylcysteine synthetase), and NQO1 (NAD(P)H dehydrogenase [quinone] 1) in the liver and spleen of oxidized mice. Insect tea has a good preventive effect on D‐galactose‐induced oxidation in mice, and the effect is better than vitamin C, an antioxidant. Insect tea is rich in isochlorogenic acid A, quercetin, rutin, hesperidin, neochlorogenic acid, and cryptochlorogenic acid. The combination of these bioactive substances has good antioxidant effects. Thus, Insect tea is a functional food with a good antioxidant effect that has value for future development and utilization., Insect tea is a traditional Chinese drink that contains abundant bioactive substances. In this study, we found that Insect tea was a functional food with a good antioxidant effect that has value for future development and utilization.

Litchi grown in the upper Yangtze River region have the advantage of being late-maturing owing to the geographical location. This study aimed to evaluate the physical characteristics, nutritional values, phenolic composition and antioxidant activities of 16 litchi cultivars grown in the upper Yangtze River region. Litchi grown in this region had total soluble solid and ascorbic acid contents comparable with those of cultivars grown in other locations. The total polyphenol contents were determined using the Folin-Ciocalteu assay, and the phenolic profiles were determinded using UPLC-QqQ-MS/MS. Nine phenolic compounds were identified and quantified in this study. Naringin, rutin and p -coumaric acid were the major phenolic compounds in all the litchi cultivars. Statistical analysis of all the physiochemical results was performed using principal component analysis. Our results indicated that litchi grown in the upper Yangtze River region not only showed the late-maturity characteristic but were also good dietary sources of phenolic compounds and antioxidants. In particular, 'Fei Zi Xiao' and 'Jing Gang Hong Nuo', characterized by high polyphenol contents and high antioxidant capacities, were of superior comprehensive quality. This study provides important information for the development of late-maturing litchi industry.

Uranium and nitrate are common groundwater pollutants near in situ leach uranium mines. However, we still lack techniques that can simultaneously immobilize uranium and reduce nitrate using a single bacterial species. In this study, the potential of simultaneous uranium immobilization and nitrate reduction by a single AFODN (anaerobic Fe(II) oxidizing denitrifier), Clostridium sp. PXL2, was investigated. Clostridium sp. PXL2 showed tolerance to U(VI) concentrations varying from 4.2 µM to 42 µM. The U(VI) immobilization and nitrate reduction rates in groundwater samples inoculated with this bacterium reached up to 75.1% and 55.7%, respectively, under neutral conditions. Exposure to oxidation conditions led to further U(VI) removal but did not show any noticeable effect on nitrate reduction. The U(VI) immobilization rate reached up to 85% with an increased Fe(II) initial concentration, but this inhibited nitrate reduction. SEM (scanning electron microscopy) coupled with EDS (energy dispersive spectroscopy) showed that the U(VI) immobilization was mainly due to sorption to amorphous ferric oxides. U(VI) and nitrate bioremediation by AFODNs, including Clostridium sp. PXL2, may provide a promising method for the treatment of uranium- and nitrate-contaminated groundwater after the in situ leach mining of uranium.

Due to the rapid development of solar cells based on organic metal halide perovskite as light-absorber, the power conversion efficiency (PCE) of perovskite solar cells (PSCs) has increased from 3.8% to 25.5%, which exceeds that of silicon ones. In the meantime, the stability of the device has also made breakthrough progress. However, the degradation of perovskite is the primary reason which largely limits large-scale application. PSCs quickly chemically react with the water/oxygen in the air and decompose when exposed to ambient condition. It is urgently necessary to develop appropriate encapsulation techniques to isolate perovskite from external stimuli including moisture, oxygen and UV light to effectively extend lifetime. In this paper, the influences and physical mechanisms of the stability of perovskite are summarized. Then, the main encapsulation methods and technical approaches to improve and enhance the stability of perovskite are also reviewed, including traditional rigid-substrate-based encapsulation, thin film encapsulation, and advanced encapsulation strategies. Finally, it overviews the challenges and further development strategies faced by PSCs encapsulation techniques in practical applications.

Homocysteine (Hcy) is a non-essential amino acid produced from methionine. It has been reported that high concentrations of Hcy are related to the pathogenesis of neurodegenerative diseases and induce the disruption of the blood-brain barrier (BBB) by triggering oxidative stress and inflammation. LCZ696 is a novel antihypertensive agent that has been recently reported to possess promising anti-inflammatory properties. However, whether it has a protective effect on the BBB disruption is still unknown. For the first time, in this study, we aim to investigate whether LCZ696 exerts anti-inflammatory effects on Hcy-induced injury in brain endothelial cells and explore its neuroprotective properties. In in vivo experiments, we found that treatment with LCZ696 ameliorated oxidative stress by reducing malondialdehyde (MDA) and increasing glutathione (GSH). Furthermore, LCZ696 downregulated the excessive release of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) at mRNA and protein levels. Importantly, it reversed the disruption of the BBB induced by Hcy stimulation. In the in vitro human brain microvascular endothelial cell (HBMVEC) experiments, compared to the control, the permeability of the endothelial monolayer was significantly enlarged, the expression level of occludin declined, and Egr-1 upregulated by the introduction of Hcy, and these were all reversed by the treatment with LCZ696. Lastly, we found that the protective effects of LCZ696 against Hcy-induced reduction of occludin and hyper-permeability of the endothelial monolayer were greatly abolished by the overexpression of Egr-1. Taken together, we found that LCZ696 protected against Hcy-induced impairment of BBB integrity by increasing the expression of occludin, all mediated by the inhibition of Egr-1.

Phosphate starvation (-Pi)-induced root hair is crucial for enhancing plants' Pi absorption. Proline-rich extensin-like receptor kinase 13 (PERK13) is transcriptionally induced by -Pi and co-expressed with genes associated with root hair growth. However, how PERK13 participates in -Pi-induced root hair growth remains unclear. Here, we found that PERK13 was transcriptionally responsive to Pi, nitrogen, and iron deficiencies. Loss of PERK13 function (perk13) enhanced root hair growth under Pi/nitrogen limitation. Similar phenotype was also observed in transgenic lines overexpressing PERK13 (PERK13ox). Under -Pi, both perk13 and PERK13ox showed prolonged root hair elongation and increased reactive oxygen species (ROS). Deletion analysis showed, in PERK13ox, the extracellular domain was indispensable for PERK13 in -Pi-induced root hair growth. Different transcription profiles were observed under -Pi between perk13 and PERK13ox with the jasmonate zim-domain genes being repressed in perk13 and genes involved in cell wall remodeling being increased in PERK13ox. Taken together, we demonstrated that PERK13 participates in -Pi-induced root hair growth probably via regulating root hair elongation and the generation of ROS. Our study also suggested PERK13 probably being a vital hub coupling the environmental cues and root hair growth, and might play dual roles in -Pi-induced root hair growth via different processes.

The aim of this study was to investigate the effects of three different drying methods, freeze drying (FD), vacuum drying (VD) and oven drying (OD) on phenolic contents and antioxidant activities of litchi fruits. 20 polyphenols were exactly identified in the litchi fruits by UPLC-QqQ/MS. Significant losses were observed in the contents of total polyphenols and antioxidant activities in the dried litchi when compared with the fresh litchi. Principle component analysis indicated that there was significant difference of phenolic component between the use of thermal drying (VD and OD) and FD. Our results suggest that FD is the optimum drying method for litchi fruits considering the content of total polyphenols and antioxidant activities.

BackgroundSmall-cell lung cancer (SCLC) remains an aggressive cancer with short-term survival due to limited therapeutic options. Apatinib is a small-molecule tyrosine kinase inhibitor that selectively inhibits vascular endothelial growth factor receptor-2. This study aimed to investigate the efficacy and safety of apatinib in patients with extensive-stage (EC) SCLC who had progressed after two or three previous therapies.MethodsEligible patients were histologically confirmed ES-SCLC after two or three previous treatments, including a platinum-based regimen. Patients received apatinib at an initial dose of 500 mg once daily. The primary endpoint was the objective response rate.ResultsForty patients were enrolled. At the data cut-off time (November 15, 2018), the median follow-up was 7.4 months; no patients remained on treatment, and five were still in follow-up. An objective response was achieved in 7 of 40 patients (17.5%) in the intention-to-treat population, and 7 of 38 patients (18.4%) in the per-protocol population. The median progression-free survival and overall survival were 3.0 months and 5·8 months, respectively. The most commonly observed grade 3 or greater treatment-related adverse events were hypertension, hand–foot syndrome, increased L-gamma-glutamyltransferase.ConclusionsApatinib exhibited efficacy and an acceptable safety profile in previously heavily-treated ES-SCLC patients. Further exploration of apatinib in phase III trials is warranted.Trial registrationNCT02945852.

Research hotspots about Ce3+-Tb3+ and Ce3+-Mn2+ codoped multicolor-tunable materials, has grown rapidly and is still growing. However, few feasible strategies can be employed to achieve the multicolor-tunable luminescence under the premise of maintaining high QE. In this paper, KSrLu(PO4)2:0.25Ce presents a high luminescence efficiency with approximately 78% quantum efficiency. Furthermore, Ce3+-Tb3+ and Ce3+-Mn2+ codoped optimal samples give a green and red luminescence, which corresponds to a high 61% and 55% quantum efficiency due to a high energy transfer efficiency of Ce3+→Tb3+ and Ce3+→Mn2+. Besides, we achieved the precise adjustment of blue, green, orange and pink emissions of KSrLu(PO4)2:Ce3+, Tb3+, Mn2+ samples. Meanwhile, KSrLu(PO4)2:Ce3+, Tb3+ and KSrLu(PO4)2:Ce3+, Mn2+ phosphors present a weak thermal quenching: their integrated emission intensity at 500K still keep more than 80% of their initial intensity at room temperature. Therefore, the results might provide a novel multicolor-tunable phosphor for backlight display application.

Design and preparation of novel advanced carbon materials with unique architecture and functional groups is of great significance. Herein, a spongy acetylenic carbon material (SACM) was prepared through mechanochemical reaction of CaC2 and chlorinated rubber in a planetary ball mill at ambient temperature. Its composition and structure were characterized, and its electrochemical properties and adsorption performance for Hg2+ were studied. The SACM is composed of submicron spongy aggregates with high carbon content (81.8%) and specific area (503.9 m2·g−1), rich porosity and acetylenic groups. The SACM exhibits excellent adsorption for Hg2+ with saturated adsorption amount being 157.1 mg·g−1, which is superior to conventional carbon materials. Further, it exhibits good electrochemical performance with low equivalent series resistance (0.50 Ω), excellent cycling stability and ideal double layer capacitive behavior. This paper provides a novel and universal synthesis method of spongy carbon materials, and better results can be expected through tuning the pore structure, graphitization degree, and heteroatoms of the target carbon materials.

Dielectric barrier discharge (DBD) plasma in situ generates reactive species (e*, OH, O, H, H2O2, O3, etc.), UV irradiation and local high temperature. These physiochemical effects can non-selectively destroy aqueous organic contaminants, but the energy efficiency of DBD plasma for water decontamination can be further improved. Persulfate ( S 2 O 8 2 - , PS) and ferrous ions (Fe2+) were employed for enhancing the degradation of p-nitrophenol (PNP). An addition of both S 2 O 8 2 - ( OH and SO 4 · - through Fenton-like reactions and PS activation process. Moreover, the tertiary system of plasma/PS/Fe2+ (81.1%) presented higher PNP degradation efficiency after 50 min treatment than discharge plasma alone (34.8%) and the binary systems of plasma/PS (63.6%), plasma/Fe2+ (69.6%) and PS/Fe2+ (13.7%). However, an excessive addition of S 2 O 8 2 - (2.5–3.6 mM) and Fe2+ (36–360 µM) had unobvious or even harmful influence on PNP degradation, possibly due to the enhanced recombination or consumption of OH and SO 4 · - by radical side reactions. The effect of radical scavengers on PNP degradation indicates that OH was the most important radical species in plasma system for PNP degradation, but SO 4 · - was also very important for PNP degradation when PS was present in solution. Besides Fe2+ and the heat effect from discharge plasma, the electrons produced by electric discharge may also be one of the important contributors to the activation of PS.

Fructose as a daily sweetener is widely recognized as a risk catalyst for nonalcoholic fatty liver disease (NAFLD). The aim of current study is to evaluate the effects and molecular mechanism by which polyphenol-rich loquat fruit extract (LFP) prevents NAFLD in mice fed 30% fructose water (HF) for 8 weeks. Administration of LFP to HF-fed mice mitigated abnormal body weight, disordered lipid metabolism, oxidative stress, and inflammation through a mechanism regulated by the AKT, ChREBP/SREBP-1c, Nrf2, and TLR4/MyD88/TRIF pathways. LFP caused a significant decrease in the endotoxin content (16.67-12.7 EU/mL) in the liver of HF-fed mice. LFP not only improved HF-induced breakage of the intestinal barrier via interacting with tight junction proteins (ZO-1, occludin), mucin, and immunoreaction in the colon but also maintained normal colonic Firmicutes/Bacteroidetes ratios and the relative abundance of Veillonella in HF-fed mice. Our results suggest that LFP may serve as a nutritional agent for protecting liver in HF-fed mice.

A novel method is presented for the synthesis of Ti2SC powder by heating FeS2/Ti/TiC mixtures. The phase evolution of Ti2SC was explored in detail with X-ray diffraction, thermo-gravimetry/differential scanning calorimetry, and scanning electron microscopy analysis. It was found that FeS2 began to decompose into activated FeS and released molecular state sulphur at around 500 °C. Subsequently, FeS and Ti underwent a reaction that formed Fe and TiS at 700 °C. Finally, the reaction of both TiS and TiC to form a Ti2SC phase could be triggered at 900 °C. It was confirmed that the Ti2SC phase was obtained with only trace impurity of TiC remaining after acid treatment by microwave hybrid heating of 1.1FeS2/2Ti/2TiC mixtures at 1 300 °C for 3 min. The fine powder with a mean diameter of approximately 1 μm was mainly composed of quasi-spherical particles. Compared to the conventional method, Ti2SC powder was synthesised at a temperature some 300 °C lower and required only 10 % of the heating time.

To deal with serious environmental pollution resulting from plastic packaging materials, biodegradable films using chitosan (CS) are gaining considerable increase gradually. However, chitosan films lack important properties to meet the preserved demands. This study aimed to develop new bio-based films incorporated with carboxymethyl chitosan-ZnO (CMCS-ZnO) nanoparticles and sodium alginate (SA) to overcome the weakness of CS films. CMCS-ZnO nanoparticles were successfully synthesized in the matrix of CMCS through direct precipitation method, which showed an average diameter of 100 nm. Multilayer films with CS film as the outer layer and SA film as the inner layer were prepared by solution casting method. The addition of CMCS-ZnO nanoparticles led to enhanced tensile strength, and to better water vapor resistance. The as-prepared films exhibited distinctive antibacterial activity against S. aureus and E. coli. The results suggested that the as-prepared film is expected to be a promising material for food packaging.

The development of high drug loading and sustained release nanoparticles by simple and economical methods is significant for the treatment of diseases. Amorphous calcium carbonate (ACC) is an ideal drug delivery system because of its excellent pH responsiveness, biocompatibility and biodegradability. In this work, ACC nanoparticles were successfully precipitated in ethanol–calcium solutions using ammonium carbonate as the internal source of CO2, which exhibited a relatively narrow size distribution in range of 10–200 nm. The microscopic morphology, amorphous characters, porous structure and thermal behavior of resultants were investigated by electron microscopy, Fourier-transform infrared spectroscopy, Brunauer–Emmett–Teller and thermogravimetric analysis, respectively. In vitro drug release assay showed that ACC nanoparticles had high loading capacity of curcumin (Cur) and favorable drug release properties. Furthermore, ACC-Cur showed excellent abilities to scavenge free radicals, protect Cur stability and damage A549 cells, providing a broadened space for the applications of this material in the fields of biomedical or food.

Porphyromonas gingivalis , a clinically important oral pathogen causing periodontal disease, is difficult to culture in routine conditions. Hence, it is necessary to establish a reliable technique to detect this pathogen. Previously, our laboratory developed a new isothermal detection method, called MB-LAMP (molecular beacon-Loop-mediated isothermal amplification), which combines the advantages of LAMP and qPCR through the accurate and quantitative detection of LAMP products. This approach offers significant potential for the point-of-care detection of P. gingivalis. Here, MB-LAMP was used to detect P. gingivalis targeting a specific fragment, and the sensitivity was as high as 1.4 × 10−1 pg μL–1. The method showed no cross-reaction with 14 other bacterial pathogens. For clinical samples, this assay showed a high diagnostic sensitivity (100%) and specificity (100%), equivalent to that of real-time quantitative polymerase chain reaction (real-time qPCR). Moreover, detection with MB-LAMP was significantly faster than that with real-time qPCR, reducing the time required for clinical diagnosis. Finally, we established an absolute quantification method with MB-LAMP for P. gingivalis using pilot samples. Thus, the highly specific, sensitive, and rapid assay developed in this study makes it feasible to diagnose P. gingivalis.

Osteoarthritis (OA) is a degenerative joint disease, in which low-grade inflammation plays an important role at the initiating step. Low-doses of LPS-induced inflammation in the plasma activate chondrocytes and promote the secretion proinflammatory cytokines, leading to secondary inflammation. Blocking OA-associated TLR activation is a promising strategy for the development of suitable therapies. Here, we want to find some bacteria-derived peptides that can block TLR signaling in chondrocytes more efficiently. Based on previous studies, we screened 12 TIR domain-derived peptides for their effects on NF-кB activation induced by LPS, IL-1β or TNF-α in murine ATDC-5 cells. We evaluated their effects on LPS-induced cytokine expression and secretion. Among them, two bacteria-derived peptides, TcpC-DD and TcpB-DD, showed the most potent inhibitory activities. In comparison with TcpB-DD, TcpC-DD exhibited broader TLR-inhibitory specificity during inflammation in chondrocytes. Furthermore, both TcpC-DD and TcpB-DD displayed strong inhibition of LPS- and IL-1β-induced catabolic reactions in chondrocytes. However, only TcpC-DD exhibited obvious suppression of TNF-α-induced catabolism. In conclusion, we identified two novel inhibitory peptides that modulate catabolism in chondrocytes and innate immune responses, and these peptides could be used to develop novel therapeutic strategies for OA.

The current study evaluated the effects of vacuum drying on the whole polyphenol profile of loquat fruit, including extractive and nonextractive polyphenols. Absorbance analysis determined that total polyphenol content and antioxidant levels were higher in loquat fruit vacuum dried at 140 °C than in loquat fruit vacuum dried at 70 °C. The results of ultra-HPLC-triple quadruple mass spectrum analysis showed that 15 phenolic acids and 17 flavonoids were found in dried loquat fruit. Multivariate integrative (MINT) sparse partial least square-discriminant analysis showed that vacuum drying affects the polyphenol profile of loquat fruit. Co-analysis of principal component analysis, partial least square-discriminant analysis, and orthometric partial least square-discriminant analysis revealed that vacuum drying mainly changed the content of chlorogenic acid, cryptochlorogenic acid, protocatechuic acid, phloretin, and hesperidin in loquat fruit. Chlorogenic acid (12.020 to 39.153 µg/g d.b. [dried base weight]), the main polyphenol in dried loquat fruit, was degraded to caffeic acid (0.028 to 2.365 µg/g d.b.) and protocatechuic acid (0.014 to 18.285 µg/g d.b.) during vacuum drying. Moreover, vacuum drying also induced the isomerization of chlorogenic acid into cryptochlorogenic acid (1.628 to 12.737 µg/g d.b.). These results might be used to develop dried loquat fruit with high levels of polyphenols and antioxidant activity. PRACTICAL APPLICATION: Interests in polyphenols of loquat fruit had increased greatly because of their possible role in health benefits. This work provided a holistic insight in the effects of vacuum drying on polyphenols profile of loquat fruit. Current results have contributed to the development of vacuum-drying method, which produced loquat fruit rich in polyphenols. Furthermore, it also suggested that multivariate analysis was a feasible method to reveal the important changes of polyphenols profile during food processing.; © 2019 Institute of Food Technologists®.

Accelerating and regulating collagen formation during wound healing repair is key issues for skin regeneration. Insulin can promote the healing of damaged skin by stimulating cellular migration and angiogenesis. Here, human hair keratin-conjugated insulin was synthesized to enhance full-thickness skin regeneration based on the excellent wound healing and hemostatic effects of keratin and the collagen deposition regulation ability of insulin. The insulin-conjugated keratin (Ins-K) was synthesized through the EDC/NHS reaction, which can supply a sustained release of insulin. The Ins-K hydrogel displayed similar water absorption, porosity and rheology properties to those of the keratin hydrogel. However, the Ins-K hydrogel shows a stronger hemostatic ability than the keratin hydrogel group, with a stronger wound healing effect found for the Ins-K hydrogel in the early regeneration stage (first 2 weeks) than for the keratin hydrogel treatment, resulting in smoother skin tissues at an excision section realized by regulating transforming growth factor β1 (TGF-β1) and hydroxyproline (HYP) expression. The results demonstrate that keratin promotes hemostasis and wound healing after insulin conjugation, which highlights the potential of keratin-based materials in tissue regeneration applications.

6-Phosphogluconate dehydrogenase (6PGD) is a key enzyme that converts 6-phosphogluconate into ribulose-5-phosphate with NADP+ as cofactor in the pentose phosphate pathway (PPP). 6PGD is commonly upregulated and plays important roles in many human cancers, while the mechanism underlying such roles of 6PGD remains elusive. Here we show that upon EGFR activation, 6PGD is phosphorylated at tyrosine (Y) 481 by Src family kinase Fyn. This phosphorylation enhances 6PGD activity by increasing its binding affinity to NADP+ and therefore activates the PPP for NADPH and ribose-5-phosphate, which consequently detoxifies intracellular reactive oxygen species (ROS) and accelerates DNA synthesis. Abrogating 6PGD Y481 phosphorylation (pY481) dramatically attenuates EGF-promoted glioma cell proliferation, tumor growth and resistance to ionizing radiation. In addition, 6PGD pY481 is associated with Fyn expression, the malignancy and prognosis of human glioblastoma. These findings establish a critical role of Fyn-dependent 6PGD phosphorylation in EGF-promoted tumor growth and radiation resistance., 6-phosphogluconate dehydrogenase is commonly upregulated in cancers. Here, the authors show that activation of EGFR induces phosphorylation of this enzyme at Y481 to activate the pentose phosphate pathway, which consequently reduces ROS and accelerates DNA synthesis to promote tumor growth and radioresistance.

espanolPara este estudio, se elaboro glucomanano de konjac oxidado y degradado por ultrasonidos (U-OKGM), sometiendose a examen la actividad inmune que proporciona a los macrofagos RAW264.7. Los resultados dan cuenta de que el tratamiento por ultrasonidos demostro ser un metodo efectivo para obtener un bajo peso molecular de la OKGM; por otra parte, dicho tratamiento no modifico la estructura principal de los polisacaridos sometidos a las condiciones de prueba. Los pesos moleculares (MW) del OKGM y el U-OKGM obtenidos fueron de alrededor de 1.58 × 106 g/mol y 5.64 × 105 g/mol, respectivamente. Se constato que el U-OKGM puede inhibir la secrecion de NO y aumentar la actividad fagocitica de las celulas RAW264.7, ademas de que la cantidad de U-OKGM lograda, 0.8 mg/mL, fue significativa (p EnglishThe ultrasound-degraded oxidized konjac glucomannan (U-OKGM) was manufactured and its immune activity in RAW264.7 macrophages was examined. Results showed that ultrasound treatment, which was an effective method to obtain low molecular weight OKGM, did not change the main structure of polysaccharides under the test conditions. Molecular weights (MW) of OKGM and U-OKGM are about 1.58 × 106 g/mol and 5.64 × 105 g/mol, respectively. We showed that U-OKGM could inhibit NO secretion and augment phagocytotic activity of RAW264.7 cell, and 0.8 mg/mL U-OKGM reached significant level (p

The swollen succulent stem of tumorous stem mustard (Brassica juncea var. tumida Tsen et Lee) is the raw material in the processing of Chinese Fuling zhàcài. We found that total polyphenols and flavonoids contents of tumorous stem mustard leaf extracts (TSMLE) were significant higher than that of tumorous stem mustard stem extracts (TSMSE). The percentage of isorhamnetin 5-O-hexoside, methyl quercetin O-hexoside, and luteolin O-hexosyl-O-hexosyl-O-hexoside in TSMLE were 170, 230 and 694 times higher than that in TSMSE. TSMLE presented stronger antioxidant capacity against 2,2'-Azino-bis[3-ethylbenzothiazoline]-6-sulfonic acid cationic free radical (ABTS+) and 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radicals, and it also showed higher reducing power and ferric reducing antioxidant power. Both TSMLE and TSMSE inhibited lung carcinoma cell growth in a dose-dependent manner, while TSMLE was more effective against A549 cells than TSMSE. This is the first report indicates that tumorous stem mustard, especially its leaf, is a potential diet source in preventing of oxidative stress and cancer.

In the study, amorphous calcium phosphate (ACP) nanoparticles were prepared by the coprecipitation method to optimize release profile of curcumin (Cur) and avoid burst releases, which were used to overcome the weakness of Cur, such as poor chemical stability and bioavailability. To find the best preparation condition, the influence of reaction concentration, temperature, time and pH on crystal phase of the samples was investigated by XRD and FTIR. The results showed that amorphous calcium phosphate (ACP) was obtained, when pH value and the concentration of PO43− were 8 and 0.024 mM, prepared at 30 °C for 10 min. In vitro drug release assay, ACP nanoparticles showed high loading capacity of Cur and favorable pH-responsive drug release properties. Furthermore, the Cur-load ACP nanoparticles showed an excellent ability to scavenge free radical and damage A549 cells, resulting in a high antioxidant properties and low cell viability. Therefore, the as-prepared nanoparticles have promising applications in the food and biomedical fields.

Phosphate-stabilized amorphous calcium carbonate (ACCP) has excellent biocompatibility, bioactivity, and biodegradability, and can be easily synthesized and stored. However, unmodified ACCP, as a controlled drug release carrier, decomposes rapidly in an acidic environment and highly depends on the system's pH value, which can not meet the need for long-term release of active substances, thus limiting its application scope. To realize the specific pH responsiveness of ACCP nanoparticles, we designed and synthesized monodisperse sodium alginate/ACCP (Alginate/ACCP) composite nanoparticles in this paper. After ultrasonic treatment, nanoparticles with an average particle size less than 200 nm could form stable water dispersion that could be dispersed for up to 10 d. Based on the specific pH sensitivity of sodium alginate, the drug-controlled release performance of composite nanoparticles and the therapeutic effect of drug-loaded nanoparticles on A549 cancer cells were studied. The results indicated that under the same pH condition, the curcumin (Cur) release rate of composite nanoparticles gradually decreased with sodium alginate addition. When the dosage of sodium alginate was 1.0 mg ml-1, the cumulative drug release rate of nanoparticles in 40 h was only about 35%. Besides, the drug-loaded nanoparticles showed the excellent killing ability of cancer cells, and the survival rate of cancer cells decreased in a concentration-dependent manner. Therefore, through reasonable optimization design, we can synthesize composite nanoparticles with excellent sustained-release properties to provide a new strategy for cancer cells' long-term treatment.

Recent studies have been found that polyphenols from plums fruits can inhibit the proliferation of multiple cancer cells, while the molecular mechanism was unclear. This study aimed to investigate the molecular mechanism underlying the pro-apoptotic effect of purified plum polyphenols (PPP) on human lung cancer A549 cells. Quercitrin (quercetin-3-O-glucoside, 814.19 ± 40.71 mg/g) was identified as the primary polyphenol in PPP via ultra high-performance liquid chromatography coupled with triple quadrupole mass spectrometry (UHPLC-QqQ-MS/MS). PPP showed a strong capacity for inhibiting the proliferation of the A549 cells by inducing apoptosis, which was reflected by an increase in the Bax/Bcl-2 ratio. Additionally, the inhibitory rate of PPP on the A549 cells were higher than that of vitamin C when the treatment dose exceeded 160 μg/mL. Transcriptome analysis suggested that PPP-induced apoptosis was closely associated with regulating the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT)/forkhead box protein O 1 (FOXO1) pathway in the A549 cells. Subsequently, as an activator of AKT, SC79 was applied to confirm that the inhibition of AKT phosphorylation play an important role in the PPP-induced apoptosis of the A549 cells. These results illustrated the potential of PPP as a dietary compound for the prevention of cancer or for use during chemotherapy.

Recently, the emergence of immunotherapy has revolutionized traditional tumour treatment. However, effective treatments for patients exhibiting αPD-1 resistance are still lacking. In our study, a combination of cytosine–phosphate–guanine oligodeoxynucleotides (CpG-ODNs), anti-OX40 and cyclic guanosine monophosphate–adenosine monophosphate (cGAMP) injection in situ systematically generated a robust antitumour immune response in TC1 and B16 cells, which are αPD-1-resistant malignancies. More precisely, this method activates both adaptive and innate immunity. Additionally, in situ vaccination with CpG/αOX40/cGAMP fully activates the production of cytokines. However, the combination of αPD-1 does not improve the efficacy of triple therapy, prompting further questions. Collectively, the combination of CpG/αOX40/cGAMP causes the regression of various αPD-1-resistant tumours through the full mobilization of innate and adaptive immunity. In addition, we explored the therapeutic effect of triple therapy on the αPD-1-sensitive cell line CT26. The results showed that triple therapy could significantly enhance the therapeutic effect of αPD-1, and some mice even achieved complete tumour regression after the combined application of αPD-1 and triple treatment.