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Dasatinib is a multi-target protein tyrosine kinase inhibitor. Due to its potent inhibition of Src, Abl, the platelet-derived growth factor receptor (PDGFR) family kinases, and other oncogenic kinases, it has been investigated as a targeted therapy for a broad spectrum of cancer types. However, its efficacy has not been significantly extended beyond leukemia. The mechanism of resistance to dasatinib in a wide array of cancers is not clear. In the present study, we investigated the effect of dasatinib on hepatocellular carcinoma cell growth and explored the underlying mechanisms. Our results showed that dasatinib potently inhibited the proliferation of SNU-449 cells, but not that of other cell lines, such as SK-Hep-1, even though it inhibited the phosphorylation of Src on both negative and positive regulation sites in all these cells. Dasatinib activated the phosphoinositide-dependent protein kinase1 (PDK1)/protein kinase B (Akt)/mammalian target of rapamycin (mTOR) signaling pathway in SK-Hep-1 cells, but not in SNU-449 cells. Blocking the Akt/mTOR signaling pathway strongly promoted the efficacy of dasatinib in SK-Hep-1 cells. In SNU-449 cells, dasatinib promoted apoptosis and the cleavage of caspase-3 and caspase-7, induced cell cycle arrest in the G1 phase, and inhibited the expression of Cyclin-dependent kinase (CDK4)/6/CyclinD1 complex. These findings demonstrate that dasatinib exerts its anti-proliferative effect on hepatocellular cell proliferation by blocking the Src family kinases; however, it causes Akt activation, which compromises dasatinib as an anti-cancer drug.

BACKGROUND Monacolin K, an important secondary metabolite of Monascus, possesses a cholesterol-lowering effect and is widely used in the manufacture of antihypertensive drugs. In the present study, we constructed an extractive fermentation system by adding non-ionic surfactant and acquired a high monacolin K yield. The mechanism was determined by examining both cell morphology and the transcription levels of the related mokA-I genes in the monacolin K biosynthetic gene cluster. RESULTS The monacolin K yield was effectively increased to 539.59 mg L-1 during extraction, which was an increase of 386.16% compared to that in the control group fermentation. The non-ionic surfactant showed good biocompatibility with Monascus. Electron scanning microscopy revealed alterations in the morphology of Monascus. The loosened mycelial structure and increased number of cell surface wrinkles were found to be related to the increased cell-membrane permeability and extracellular accumulation of monacolin K. Gene expression levels were measured via a quantitative reverse transciptase-polymerase chain reaction. By contrast, in the control group, mokA, mokB, mokC, mokD and mokF showed higher-level and longer-term expression in the extractive fermentation group, whereas mokE and mokG did not present a similar trend. The expression levels of mokH and mokI, encoding a transcription factor and efflux pump, respectively, were also higher than the control levels. CONCLUSION The addition of a non-ionic surfactant to Monascus fermentation effectively increases the yield of monacolin K by transforming the fungus morphology and promoting the expression of monacolin K biosynthesis genes. © 2021 Society of Chemical Industry.

β1,4-GalT1 is a type II membrane glycosyltransferase. It catalyzes the production of lactose in the lactating mammary gland and is supposedly also involved in the galactosylation of terminal GlcNAc of complex-type N-glycans. In-vitro studies of the bovine β4Gal-T1 homolog showed that replacing a single residue of tyrosine with leucine at position 289 alters the donor substrate specificity from UDP-Gal to UDP-N-acetyl-galactosamine (UDP-GalNAc). The effect of this peculiar change in β1,4GalT1 specificity was investigated in-vivo, by generating biallelic Tyr286Leu β1,4GalT1 mice using CRISPR/Cas9 and crossbreeding. Mice bearing this mutation showed no appreciable defects when compared to wild-type mice, with the exception of biallelic female B4GALT1 mutant mice, which were unable to produce milk. The detailed comparison of wild-type and mutant mice derived from liver, kidney, spleen, and intestinal tissues showed only small differences in their N-glycan pattern. Comparable N-glycosylation was also observed in HEK 293 wild-type and knock-out B4GALT1 cells. Remarkably and in contrast to the other analyzed tissue samples, sialylation and galactosylation of serum N-glycans of biallelic Tyr286Leu GalT1 mice almost disappeared completely. These results suggest that β1,4GalT1 plays a special role in the synthesis of serum N-glycans. The herein described Tyr286Leu β1,4GalT1 mutant mouse model may, therefore, prove useful in the investigation of the mechanism which regulates tissue-dependent galactosylation.

Monascus is a filamentous fungus that produces several secondary metabolites. Here, we investigated the effects of the global regulator LaeA on the synthesis of pigments and monacolin K in Monascus purpureus with spectrophotometer and HPLC methods. The LaeA gene was isolated from M. purpureus M1 to create an overexpression construct. An LaeA-overexpressing strain L3 was with 48.6% higher monacolin K production than the M1 strain. The L3 strain also produced higher Monascus pigments than the M1 strain. SEM showed that LaeA overexpression resulted in altered mycelial morphology. Compared with the M1 strain, the L3 strain expressed higher levels of monacolin K synthesis-related genes mokA, mokB, mokE, and mokH. Overall, these results suggest that LaeA plays a role in regulating the production of secondary metabolites and mycelial growth in Monascus. This study provides important insights into the mechanisms underlying the effects of the LaeA gene on the secondary metabolites of M. purpureus.

Sarcodon aspratus is a popular edible fungus that has commonly been used as a functional food in China and other Asian countries. This study is conducted to examine the potential health benefits of Sarcodon aspratus polysaccharides (SATPs), on obesity and related metabolism disorders. C57BL/6J mice were fed with a high-fat diet (HFD) and supplemented with SATPs (100-400 mg kg-1) for 14 weeks. The results indicated that SATP treatment markedly reduced HFD-induced body weight gain and fat accumulation in a dose-dependent manner. SATPs could improve lipid homeostasis and glucose tolerance in HFD-fed mice. Furthermore, SATP intervention significantly attenuated hepatic steatosis, liver oxidative stress and inflammation. Additionally, we detected the macrophage and mRNA levels of lipogenesis markers in epididymal adipose tissues, and the results revealed that SATPs exerted inhibitory effects on the activation of immune cells and adipocyte differentiation in adipose tissues. High-throughput pyrosequencing of 16S rRNA suggested that SATP intervention was able to down-regulate the Firmicutes-to-Bacteroidetes ratio, and also increase the relative abundance of Lactobacillus, Bacteroides and Akkermansia in mice with HFD challenge. Taken together, SATPs showed ameliorative effects on hepatic steatosis, inflammation and adipocyte differentiation in HFD-fed mice. Notably, SATPs could modulate HFD-induced dysbiosis of gut microbiota. Thus, they might be a potential health supplement or prebiotic in the prevention of obesity and related metabolic disorders.

A bio-barcode immunoassay based on droplet digital polymerase chain reaction (ddPCR) was developed to simultaneously quantify triazophos, parathion, and chlorpyrifos in apple, cucumber, cabbage, and pear. Three gold nanoparticle (AuNP) probes and magnetic nanoparticle (MNP) probes were prepared, binding through their antibodies with the three pesticides in the same tube. Three groups of primers, probes, templates, and three antibodies were designed to ensure the specificity of the method. Under the optimal conditions, the detection limits (expressed as IC10) of triazophos, parathion, and chlorpyrifos were 0.22, 0.45, and 4.49 ng mL–1, respectively. The linear ranges were 0.01–20, 0.1–100, and 0.1–500 ng mL–1, and the correlation coefficients (R2) were 0.9661, 0.9834, and 0.9612, respectively. The recoveries and relative standard deviations (RSDs) were in the ranges of 75.5–98.9 and 8.3–16.7%. This study provides the first insights into the ddPCR for the determination of organophosphate pesticides. It also laid the foundation for high-throughput detection of other small molecules., This study was financially supported by the National Key Research Program of China (No. 2019YFC1604503), the NIEHS Superfund Research Program (No. P42 ES04699), the Central Public-interest Scientific Institution Basal Research Fund for the Chinese Academy of Agricultural Sciences (No. Y2021PT05), and the Ningbo Innovation Project for Agro-Products Quality and Safety (No. 2019CXGC007).

Extensive application of pesticides has caused a lot of environmental pollution and health problems, prompting the development of highly efficient and lowly toxic pesticide formulations. Here, buprofezin (BPF)-loaded poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (P(HB-HH)) microspheres were prepared by O/W emulsion/solvent evaporation method. Under optimal conditions (P(HB-HH) 7.07% (w/v) and PVA 1.84% (w/v)), the spherical and monodispersed microspheres were obtained. The average particle size, pesticide loading (PL), and encapsulation efficiency (EE) of the optimized microspheres were 1.2 μm, 15.68%, and 78%, respectively. Release of 80% BPF from the microspheres in pH 5 (192 h) was faster than that in pH 7 (228 h) and 8 (204 h). Moreover, kinetic analysis indicated that BPF release behaved in a non-Fickian diffusion manner (0.43

A rapid, ultra-sensitive and simple fluorescence immunoassay (FLISA) on CdSe/ZnS Quantum dots (QDs) for recognition pesticide triazophos have been established. In this assay, the immunoassay uses (CdSe/ZnS) Quantum dots (QDs) as a probe that carries monoclonal antibody (mAb). The free triazophos and OVA-haptens were competed for binding the mAb on the surface of the QDs. Therefore, the concentration of TRIAZ can get by calculation of the detection of fluorescence value. The assay has a linear response in the 10 ng L−1–25 μg L−1 TRIAZ concentration range, and limit of detection (IC10) was 0.508 ng L−1. The recovery rate in the case of spiked samples ranges from 82.6% to 96.6%, and the RSD is

It is well-known that the high-molecular-weight polycyclic aromatic hydrocarbons (HMW-PAHs) are difficult to degrade, and they can cause tremendous damaging effects in environment. In this article,...

Crop domestication, which gives rise to a number of desirable agronomic traits, represents a typical model system of plant evolution. Numerous genomic evidence has proven that noncoding RNAs such as microRNAs and phasiRNAs, as well as protein-coding genes, are selected during crop domestication. However, limited data shows plant long noncoding RNAs (lncRNAs) are also involved in this biological process. In this study, we performed strand-specific RNA sequencing of cultivated rice Oryza sativa ssp. japonica and O. sativa ssp. indica, and their wild progenitor O. rufipogon. We identified a total of 8528 lncRNAs, including 4072 lncRNAs in O. rufipogon, 2091 lncRNAs in japonica rice, and 2365 lncRNAs in indica rice. The lncRNAs expressed in wild rice were revealed to be shorter in length and had fewer exon numbers when compared with lncRNAs from cultivated rice. We also identified a number of conserved lncRNAs in the wild and cultivated rice. The functional study demonstrated that several of these conserved lncRNAs are associated with domestication-related traits in rice. Our findings revealed the feature and conservation of lncRNAs during rice domestication and will further promote functional studies of lncRNAs in rice.

Endoplasmic reticulum (ER) stress is associated with acute kidney injury (AKI) caused by various mechanisms, including antibiotics, non-steroidal anti-inflammatory drugs, cisplatin, and radiocontrast. Tunicamycin (TM) is a nucleoside antibiotic that induces ER stress and is a commonly used model of AKI. 4-phenylbutyrate (4-PBA) is a chemical chaperone and histone deacetylase (HDAC) inhibitor and has been shown to protect the kidney from ER stress, apoptosis, and structural damage in a tunicamycin model of AKI. The renal protection provided by 4-PBA is attributed to its ability to prevent misfolded protein aggregation and inhibit ER stress; however, the HDAC inhibitor effects of 4-PBA have not been examined in the TM-induced model of AKI. As such, the main objective of this study was to determine if histone hyperacetylation provides any protective effects against TM-mediated AKI. The FDA-approved HDAC inhibitor vorinostat was used, as it has no ER stress inhibitory effects and therefore the histone hyperacetylation properties alone could be investigated. In vitro work demonstrated that vorinostat inhibited histone deacetylation in cultured proximal tubular cells but did not prevent ER stress or protein aggregation induced by TM. Vorinostat induced a significant increase in cell death, and exacerbated TM-mediated total cell death and apoptotic cell death. Wild type male mice were treated with TM (0.5 mg/kg, intraperitoneal injection), with or without vorinostat (50 mg/kg/day) or 4-PBA (1 g/kg/day). Mice treated with 4-PBA or vorinostat exhibited similar levels of histone hyperacetylation. Expression of the pro-apoptotic protein CHOP was induced with TM, and not inhibited by vorinostat. Further, vorinostat did not prevent any renal damage or decline in renal function caused by tunicamycin. These data suggest that the protective mechanisms found by 4-PBA are primarily due to its molecular chaperone properties, and the HDAC inhibitors used did not provide any protection against renal injury caused by ER stress.

Monacolin K is a secondary metabolite produced by Monascus with beneficial effects on health, including the ability to lower cholesterol. We previously showed that the yield of monacolin K was significantly improved when glutamic acid was added to the fermentation broth of Monascus purpureus M1. In this study, we analyzed M. purpureus in media with and without glutamic acid supplementation using a metabolomic profiling approach to identify key metabolites and metabolic pathway differences. A total of 817 differentially expressed metabolites were identified between the two fermentation broths on day 8 of fermentation. Pathway analysis of these metabolites using the KEGG database indicated overrepresentation of the citric acid cycle; biotin metabolism; and alanine, aspartate, and glutamate metabolic pathways. Six differentially expressed metabolites were found to be related to the citric acid cycle. The effect of citric acid as an exogenous additive on the synthesis of monacolin K was examined. These results provide technical support and a theoretical basis for further studies of the metabolic regulatory mechanisms underlying the beneficial effects of monacolin K and medium optimization, as well as genetic engineering of Monascus M1 for efficient monacolin K production.

Through a screening process, we previously revealed that the addition of arginine to Monascus fermentation broth significantly increased Monacolin K production, with 0.6% arginine showing the best effects. Herein, we added 0.6% arginine to further detect the effect of arginine on secondary metabolites and mycelium morphology of Monascus purpureus. When M. purpureus was grown in a medium supplemented with arginine, the Monacolin K yield increased from 94.6 to 228.4 mg/L, indicating an increase of approximately 2.4-fold. Further, there were significant increases in Monascus pigments. Scanning electron microscopy of Monascus mycelium morphology showed that arginine significantly altered the mycelial morphology and enhanced the extracellular secretion of Monacolin K. RT-qPCR showed that after the addition of arginine, expression of all the mokA-mokI genes were upregulated. These results indicate that arginine could increase the yield of Monacolin K, providing a basis for further analysis of Monacolin K biosynthesis.

Background: Ischemia-derived exosomes can restrict excessive autophagy by transferring microRNA-30a (miR30a) to cells. Reports have confirmed that epigallocatechin gallate (EGCG) alleviates acute myocardial infarction (AMI) by regulating autophagy; however, research evaluating the communication with cardiomyocytes and exosomes is lacking. This study aimed to explore whether exosomes derived from EGCG-treated cardiomyocytes mitigated AMI by adjusting miR30a to inactivate apoptosis and autophagy. Methods: Exosomes were extracted from cardiomyocytes, cultured either in control or AMI condition, with or without EGCG pretreatment. The exosome characteristics were analyzed by nanoparticle tracking analyses (NTA) and transmission electron microscopy (TEM). The change in miR30a in cells and exosomes was demonstrated by qRT-PCR. H9c2 or stable miR30a knockdown (miR30aKD) cell lines were incubated with exosomes derived from EGCG-treated cardiomyocytes in vitro or in vivo. The effect of EGCG and exosomes on I/R-induced cardiomyocyte apoptosis and autophagy was assessed. Results: EGCG improved the activity of cardiomyocytes, and increased average diameter, concentration, miR30a mRNA level, and specific protein expression in AMI-derived exosomes produced by cardiomyocytes. Moreover, the coincubation of AMI cells with EGCG or exosomes derived from EGCG-treated cardiomyocytes attenuated cardiomyocyte apoptosis and autophagy. Conclusions: The findings showed that EGCG upregulates miR30a, which was efficiently transferred via exosomes between cardiomyocytes, thereby contributing to the suppression of apoptosis and autophagy. By focusing on the cardiomyocyte microenvironment, we identified a new target of EGCG alleviating AMI by regulating apoptosis and autophagy.

Monacolin K is a secondary metabolite of Monascus and is known to decrease cholesterol levels in humans. There are 9 genes (mokA-mokI) controlling its biosynthesis, of which mokH is thought to act as a pathway-specific regulator. In this study, the Monascus purpureus M1 strain was compared with mokH gene deletion strains (△H1) and overexpression strains (H7). The monacolin K yields in the △H1 strain were reduced by 52.05 %, and increased in the H7 strain by 82 %. The mycelium samples of the M1, △H1, and H7 strains were found to vary with scanning electron microscopy. Compared to the M1 strain, some mycelium of the △H1 strain showed obvious folding and expansion, while the mycelium of the H7 strain was fuller. Besides, these results indicate that the mokH gene can increase the yield of monacolin K by regulating the expression level of mokA-mokI genes, and influence the production of Monascus pigment. The study is the first to combine deletion and overexpression techniques to further verify the mokH gene and get the desired results in M. purpureus.

The role of antibiotics commonly used in fish culture activities in methylmercury (MeHg) formation in mariculture sediments (MS) and in reference sediments (RS) was studied using simulation microcosms. MS and RS were split into three equal batches. Two batches were spiked with Hg(NO3)2 aqueous solution at levels of 2 and 8 mg kg−1 (dry weight basis) and the remainder served as a control batch. Tetracycline (TC) and oxytetracycline (OTC) (2.5 g and 10 g of each) were added to each treatment. Sediment THg concentration decreased during the culture period possibly due to complexation of Hg with the antibiotics resulting in the dissolution of Hg compounds from the sediment. More importantly, the MeHg concentration increased after 32 days together with a decrease in the concentrations of the antibiotics in the sediment. The complexation of TC or OTC with Hg resulted in the transport of electrons from TC or OTC to Hg2+ due to the high electronegativity of Hg2+. Subsequently, Hg2+ was reduced to Hg° which reacted with CH3+ derived from TC or OTC. The use of antibiotics may therefore promote the formation of MeHg in sediments.

As a type of functional food additive, phycocyanin is shown to have a potential antineoplastic property. However, its underlying anticancer mechanism in melanoma cells remains unknown. We previously reported a 35S in vivo/vitro labeling analysis for dynamic proteomic (SiLAD) technology. It could exclusively detect protein synthesis rates via pulse labeling of newly expressed proteins by 35S, providing a high time-resolution method for analysis of protein variations. In the present study, we performed a time course analysis in A375 melanoma cells after phycocyanin treatment using SiLAD. Protein expression velocities were specifically visualized and their regulation modes were dynamically traced. Strikingly, novel protein synthesis patterns were discovered in the early phase of phycocyanin treatment, suggesting a possible mechanism of phycocyanin regulation. Furthermore, network analysis and phenotype experiments demonstrated that GRB2-ERK1/2 pathway was involved in phycocyanin-mediated regulation process and responsible for the proliferation suppression of melanoma cell, which could be a therapeutic target for malignant melanoma.

5-Fluorouracil (5-Fu) is an effective anticarcinogenic agent, however, continuous use of 5-Fu may cause severe side effects. The goal of this study was to investigate the effectiveness of Sarcodon aspratus polysaccharides (SATP) in alleviating 5-Fu-induced toxicity in Lewis tumor-bearing mice. Lewis tumor-bearing mice were treated with saline, SATP, 5-Fu or 5-Fu + SATP. The results indicated that compared to the 5-Fu group, the 5-Fu + SATP group showed effective amelioration of the liver, kidney and small intestine injury caused by 5-Fu and decreases in the levels of related biochemical indicators, such as aspartate aminotransferase (AST), alanine aminotransferase (ALT) and urea nitrogen (BUN). Additionally, the combination therapy enhanced the quality of life and immune organ indexes of mice. Further mechanistic studies indicated that the 5-Fu + SATP group showed a decrease in hepatotoxicity caused by 5-Fu via a reduction in the levels of interleukin-1β (IL-1β), an increase in the expression of Bcl-2 and decreases in the expression of p-p38, p-JNK and Bax. Collectively, the results indicated that SATP could significantly alleviate the toxicity of 5-Fu in Lewis tumor-bearing mice and showed the hepatoprotective capability of SATP via its effect on the expression levels of inflammatory factors and components of the MAPK/P38/JNK pathway, which shows that it may be a potential adjuvant for the chemotherapeutic drug 5-Fu in cancer treatment.

The role of bamboo forest in soil Hg accumulation and emissions was evaluated by analyzing Hg concentration in soil and plant samples as well as Hg flux between soil and air. THg concentrations in soil samples ranged widely from 28.5 to 860 ng g−1 with a mean value of 153 ± 17.3 ng g−1. Methylmercury concentrations in soil samples from forest soil (FS, 0.94 ± 0.20 ng g−1) were significantly higher (p

The small molecule compounds like pesticide, veterinary drug, bio-toxin, and heavy metal are widely found in animals, plants, soil, etc. Excessive compounds residues will have a bad influence on human health and the environment. Thus, it is extremely urgent that can detect the small molecules simultaneously. At present, many researches of simultaneous detection for small molecules using the method of an immunoassays have been reported thanks to its advantages of fast speed, simple operation, and high specificity. The small molecules have only one antigenic determinant, so the competitive immunoassay is the main method for small molecule compounds detection. In this paper, our main job is to describe the development of immunoassay for multi-residue detection of small molecule compounds and introduce three ways to complete the analysis of multi-residue immunoassay of small molecule compounds. We also summarize deficiencies and make an expectation of the immunoassays.

A copper complex with V-shaped ligands, [(L1)Cu2Cl2] n (1), (L1 = 4-(1H-imidazol-1-yl)phenyl) methanone) has been synthesized and characterized by IR spectroscopy, elemental analyses and singlecrystal X-ray diffraction. The crystal of 1 is monoclinic, sp. gr. C2/c with the unit cell parameters a = 17.9496(13), b = 15.3440(13), c = 7.2983(7) A, and β = 112.875(6)°, Z = 4, R1 = 0.0681, and wR2 = 0.1736 (I > 2 σ). The solid state structure of 1 consists of 2D metal aromatic chloride layers, which are propagating along the bc plane to form a 3D network through hydrogen bonds.

Objective To determine whether genetic polymorphisms in the uridine diphosphate-glucuronosyltransferase 1A ( UGT1A) and the C-C motif chemokine receptor 5 ( CCR5) genes are associated with hepatitis B virus (HBV) infection in Yi, Yao and Han ethnic groups in the Guizhou Province of China. Methods The study enrolled subjects with and without HBV infection. Whole blood was used for DNA genotyping using standard techniques. The study determined the frequencies of several polymorphic alleles ( UGT1A6 [rs2070959], UGT1A1 [rs8175347], CCR5-59029 [rs1799987] and CCR5Δ32 [rs333]) and then characterized their relationship with HBV infection. Results A total of 404 subjects were enrolled in the study: 138 from the Yao group, 101 from the Yi group and 165 from the Han group. There was a significant difference in the frequency of UGT1A1 rs8175347 polymorphisms among the three groups. The rates of 7TA carriers of UGT1A1 rs8175347 in all three groups were significantly higher than the other genotypes. Individuals with genotype AA of UGT1A6 rs2070959 in the Yi group had a higher risk for HBV infection than in the Yao and Han groups. The frequency of genotype GG in CCR5-59029 in the Yao group was significantly higher than in the Yi group. The genotypes of CCR5Δ32 were not associated with HBV infection. Conclusion These findings provide genetic and epidemiological evidence for an association of UGT1A and CCR5-59029 polymorphisms with HBV infection in Chinese Yi and Yao populations.

An Eulerian-Lagrangian simulation of the thermobaric explosion and the propagating detonation waves with aluminum particle combustion in a closed chamber is carried out and validated very favorably with the pressure histories measured experimentally. A novel numerical framework based on the space-time conservation-element and solution-element method (CE/SE) was employed to simulate the complex detonating flow with detailed chemistry. A discrete particle technique was adopted with either a mono-dispersed or poly-dispersed particle distribution. The results between the experiment and simulation on pressure histories and corresponding impulse forces are compared and showed good agreement. In addition, the effects of metal combustion on afterburning were discussed in detail in terms of the reaction mechanisms and particle dynamics.

The use of the existing endogenous neural progenitor cells (NPCs) in the brains of adult mammalian animals is challenging for cell therapy in treating Parkinson's disease (PD). Previous studies have indicated that there is a low level of neurogenesis in the substantia nigra (SN) of adult mice. To assess the regenerative/neurogenic capacity of NPCs following an intranigral injection of 6-hydroxydopamine (6-OHDA), the proliferation and differentiation of subventricular zone (SVZ)- and midbrain-derived NPCs were investigated, and the origin of SN newborn dopaminergic neurons was traced by using Nestin-CreERTM::ROSA26-LacZ mice and constructing a plasmid CD133-Promoter2-Cre. Our results showed that an intranigral injection of 6-OHDA-induced loss of dopaminergic neurons produced a significant increase in the SVZ-derived NPCs of the third ventricle (3V), cerebral aqueduct (Aq), and their surrounding regions. The SN newly generated dopaminergic neurons might contribute a little to an incomplete recovery of the nigrostriatal system. In addition, we found that SN newborn dopaminergic neurons were mainly derived from the migration and differentiation of the NPCs in the 3V- and Aq-SVZ and their adjacent regions. Thus, it will become an ideal strategy to treat PD by promoting the proliferation and differentiation of endogenous NPCs.

There is increasing interest for chemiluminescence (CL) detection with the characteristics of simplicity, low cost and high sensitivity, especially wide application of enhancersin CL detection to increased signals, prolonged luminescence time and enhanced intensity. In this review, the applications of primary enhancers and secondary enhancers were mainly described in the horseradish peroxidase–luminol system of CL enzyme immunoassay for light delay in the course of the reaction with improved sensitivity. The present review on enhancers covers the papers since 1983. Future research needs to develop novel enhancers with less interference and better performance. With wide utilization of enhancers in the CL system, the CL immunodetection technology showed a good potential and wide development prospects in food and medicine fields.

Surface-enhanced Raman scattering (SERS) has been widely used in the detection of targets and strongly depends on the interaction and the distance between the targets and nanoparticles. Herein, metal-organic frameworks (MOFs) were first easily synthesized on a large scale via a water bath method, especially Uio-66 and Uio-67. MOFs embedded with gold nanoparticles (AuNPs) for SERS enhancement were successfully fabricated via an impregnation strategy. The synthesized AuNPs/MOF-199, AuNPs/Uio-66, and AuNPs/Uio-67 composites, with LSPR properties and high adsorption capability of MOFs to preconcentrate the analytes close to the surface of the AuNPs, exhibited excellent SERS activity. The effects of the reducing concentrations of sodium citrate on the SERS activity, and the stability and reproducibility of the AuNP/MOFs have been discussed via the detection of acetamiprid. The SERS intensity enhanced by the composites was retained for more than 40 days under ambient conditions with the reducing concentrations of sodium citrate at 0.16%, 0.20%, and 0.16%. The limits of detection with the signal/noise ratio higher than 3 at the characteristic peak 632 cm-1 were 0.02 μM, 0.009 μM, and 0.02 μM for acetamiprid. Most interestingly, the AuNP/MOF-199 composites, whose morphology was long tube sheet, exhibited excellent SERS activity. These novel composites with high sensitivity, stability, and reproducibility provide a new route for the detection of pesticides via the SERS technology.

Parkinson's disease (PD) is a neurodegenerative disease caused by a gradual loss of midbrain dopaminergic (mDA) neurons in the substantia nigra pars compacta (SNpc) during aging. 1-Methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP) is one of the neurotoxins used widely to induce PD-like symptoms in PD animal models, including rodents and non-human primates. It has been reported that deletion of autophagy-related gene 7 (Atg7) in the brain results in a reduction of mDA neurons in adulthood. In this study, we used tyrosine hydroxylase (TH)-Cre mice to generate conditional knockout (CKO) mice with the specific deletion of Atg7 in mDA neurons. Consistent with previous reports, adult Atg7 CKO mice contained fewer TH-positive mDA neurons compared with wild-type (WT) controls. TH-expressing neurons containing puncta-like structures with p62 and ubiquitin immunoreactivity were observed in the midbrain of Atg7 CKO mice but were not detected in control mice. However, MPTP-induced loss of mDA neurons was not observed in Atg7 CKO mice. Our results indicate that Atg7-involved autophagy is required not only for the survival of mDA neurons in the mouse brain, but also for MPTP-induced mDA neuron degeneration.

An ultrasensitive bio-barcode competitive immunoassay method based on droplet digital polymerase chain reaction (ddPCR) was developed for the determination of triazophos. Gold nanoparticles (AuNPs) were coated with monoclonal antibodies (mAbs) and complementary double-stranded DNA (dsDNA), which included bio-barcode DNA and thiol-capped DNA. Magnetic nanoparticle (MNP) probes were constructed by modifying the MNPs with ovalbumin-hapten conjugates (OVA-hapten). The target pesticide and OVA-hapten on the surface of the MNP probes competed with the AuNP probes simultaneously, and then the bio-barcode DNA was released for quantification by ddPCR. The concentration of released DNA was inversely proportional to the concentration of pesticide to be tested. Under the optimum conditions, the competitive immunoassay exhibited a wide linear range of 0.01-20 ng/mL and a low detection limit of 0.002 ng/mL. Spike recovery tests were carried out using apple, rice, cabbage, and cucumber samples to verify the feasibility of the method. The recovery and relative standard deviations (RSDs) of the technique ranged from 76.9 to 94.4% and from 10.8 to 19.9%, respectively. To further validate the results, a linear correlation analysis was performed between the proposed method and liquid chromatography-tandem mass spectrometry (LC-MS/MS). Consequently, the bio-barcode immunoassay based on nanoparticles and ddPCR, an ultrasensitive method, showed great potential for the determination of target pesticides in real samples.

Chan Zhang,1 Ronggan Liang,2 Xiaowen Gan,2 Xiufang Yang,2 LingLin Chen,2 Jie Jian2 1Department of Pharmacology, Xiangya Hospital, Central South University, Changsha, Hunan, People’s Republic of China; 2Department of Pharmacology, Guilin Medical University, Guilin, Guangxi, People’s Republic of ChinaCorrespondence: Jie JianDepartment of Pharmacology, Guilin Medical University, 109, North 2nd Huancheng Road, Guilin, Guangxi 541004, People’s Republic of ChinaTel +86 183 7833 0768Email 251181281@qq.comBackground/Aims: Epigallocatechin gallate (EGCG) has established protective actions against myocardial ischemia/reperfusion (I/R) injury by regulating autophagy. However, little is known about the mechanisms of EGCG in posttranscriptional regulation in the process of cardioprotection. Here we studied whether microRNAs play a role in EGCG-induced cardioprotection.Methods: The myocardial I/R injury in vitro and in vivo model were made, with or without EGCG pretreatment. The upregulation and silencing of microRNA-384-5p (miR-384) and Beclin-1 in H9c2 cell lines were established. Rats were transfected with miR-384 specific shRNA. Dual-luciferase reporter gene assay was conducted to verify the relationship between miR-384 and Beclin-1. TTC staining was performed to analyze the area of myocardial infarct size. Cell viability was monitored by cell counting kit-8 (CCK-8). The release of cardiac troponin-I (cTnI) was examined by ELISA. The levels of autophagy-related genes or proteins expression were evaluated by qRT-PCR or Western blotting. Autophagosomes of myocardial cells were detected by transmission electron microscopy and laser scanning confocal microscope.Results: I/R increased both autophagosomes and autolysosomes, thereby increasing autophagic flux both in vitro and in vivo. Pretreatment with EGCG attenuated I/R-induced autophagic flux expression, accompanied by an increase in cell viability and a decrease in the size of myocardial infarction. MiR-384 expression was down-regulated in H9c2 cell lines when subjected to I/R, while this suppression could be reversed by EGCG pretreatment. The dual-luciferase assay verified that Beclin-1 was a target of miR-384. Both overexpression of miR-384 and knocking down of Beclin-1 significantly inhibited I/R-induced autophagy, accompanied by the activation of PI3K/Akt pathway, thus enhanced the protective effect of EGCG. However, these functions were abrogated by the PI3K inhibitor, LY294002.Conclusion: We confirmed that EGCG has a protective role in microRNA-384-mediated autophagy by targeting Beclin-1 via activating the PI3K/Akt signaling pathway. Our results unveiled a novel role of EGCG in myocardial protection, involving posttranscriptional regulation with miRNA-384.Keywords: epigallocatechin gallate, ischemia/reperfusion, autophagy, microRNA-384-5p, Beclin-1, PI3K/Akt pathway

Background Several novel immune checkpoint inhibitor (ICI)-based treatments exhibited promising survival benefits for metastatic renal cell carcinoma (mRCC), yet there is no current guidance regarding the optimum first-line regimen. We performed this network analysis to compare the efficacy and safety of all available treatments for mRCC. Methods A systematic search of literature was conducted up to April 30, 2019, and the analysis was done on a Bayesian fixed-effect model. Findings Twenty-five randomized clinical trials (RCTs) involving 13,010 patients were included in this study. The results showed that for overall survival, pembrolizumab plus axitinib (hazard ratio [HR]: 0.53; 95% credible interval [CrI]: 0.38–0.73) and nivolumab plus ipilimumab (HR: 0.63; 95% CrI: 0.50–0.79) were significantly more effective than sunitinib, and pembrolizumab plus axitinib was probably (68%) to be the best choice. For progression-free survival, cabozantinib (HR: 0.66; 95% CrI: 0.46–0.94), pembrolizumab plus axitinib (HR: 0.69; 95% CrI: 0.57–0.84), avelumab plus axitinib (HR: 0.69; 95% CrI: 0.56–0.85), nivolumab plus ipilimumab (HR: 0.82; 95% CrI: 0.68–0.99), and atezolizumab plus bevacizumab (HR: 0.86; 95% CrI: 0.74–0.99) were statistically superior to sunitinib, and cabozantinib was likely (43%) to be the preferred options. Nivolumab plus ipilimumab (OR: 0.50; 95% CrI: 0.28–0.84), and atezolizumab plus bevacizumab (OR: 0.56; 95% CrI: 0.36–0.83) were associated with significantly lower rate of high-grade adverse events than sunitinib. Interpretation Our findings demonstrate that pembrolizumab plus axitinib might be the best treatment for mRCC, while nivolumab plus ipilimumab has the most favorable balance between efficacy and acceptability, and may provide new guidance to make treatment decisions. Fund This research was supported by the Henan Provincial Scientific and Technological Research Project (Grant No. 192102310036).

Monascus purpureus is an important food and drug microbial resource through the production of a variety of secondary metabolites, including monacolin K, a well-recognized cholesterol-lowering agent. However, the high production costs and naturally low contents of monacolin K have restricted its large-scale production. Thus, in this study we sought to improve the production of monacolin K in M. purpureus through overexpression of four genes ( mokC, mokD, mokE, and mokI). Four overexpression strains were successfully constructed by protoplast electric shock conversion, which resulted in a 234.3%, 220.8%, 89.5%, and 10% increase in the yield of monacolin K, respectively. The overexpression strains showed clear changes to the mycelium surface with obvious folds and the spores with depressions, whereas the pBC5 mycelium had a fuller structure with a flatter surface. Further investigation of these strains can provide the theoretical basis and technical support for the development of functional Monascus varieties.

Excessive emission of plant nutrients (such as nitrogen and phosphorus) into the water body can induce eutrophication. Therefore, how to control eutrophic water efficiently and economically is very important. In the paper, highly efficient aerobic denitrifying phosphorus removing J16 bacteria was isolated from the activated sludge of an aerobic bioreactor in Taiyuan municipal wastewater treatment plant by using the blue–white spot screening method, an aerobic phosphorus absorption test, nitrate reduction test, nitrogen removal experiments, and plate coating and streaking methods. Through 16S rDNA gene homology comparison and physiological and biochemical identification, the J16 strain was preliminarily identified as Escherichia coli, with a sequence similarity of 99%. The 16S rDNA sequence of strain J16 was submitted to GenBank (accession number: MF667015). The effect of temperature, pH, percentage of inoculum and phosphate-P (PO43−-P) concentration on denitrification and phosphorus removal efficiency was investigated through a single-factor experiment. The optimum conditions of the J16 strain for denitrification and phosphorus removal were as follows: 30°C, neutral or weak alkaline (pH: 7.2–8), and 3% of inoculum, respectively. The denitrification and phosphorus removal efficiency of strain J16 was the highest when PO43−-P and nitrate-N(NO3−-N) concentrations were 8.9 and 69.31 mg/L, and the removal were 96.03% and 94.55%, respectively. In addition, strain J16 could reduce phosphoric acid to phosphine (PH3) and remove some phosphorus under hypoxia conditions. This is the first study to report the involvement of Escherichia coli in nitrogen and phosphorus removal under aerobic and hypoxia conditions. Based on the above results, the strain J16 can effectively remove nitrogen and phosphorus, and will be utilized in enhancing treatment of nitrogen and phosphorus-containing industrial wastewater and phosphorus reclamation.

Monascus purpureus is a traditional Chinese microbe that can be used as a medicinal herb and is edible. To improve the yield of monacolin K, we optimized the medium of M. purpureus with high-yield monacolin K strains. When high-yield strains C8, D8, E3, and I1 were grown in glutamic medium instead of the original medium, monacolin K production was increased. Among these strains, C8 exhibited the highest monacolin K production in glutamic acid medium, with levels increased 4.80-fold. RT-qPCR demonstrated that glutamic acid enhanced the expression of mokC and mokG. Observation of Monascus mycelium morphology using SEM showed that mycelia exhibited more folds, swelling, curves, and fractures. Thus, glutamic acid may promote the growth of the mycelium and appeared to increase the permeability of the cell membrane. This lays a foundation for research on the regulatory effect of glutamic acid and provides a theoretical basis for the industrialization and commercialization of Monascus.

Background: Several novel immune checkpoint inhibitor (ICI)-based treatments exhibited promising survival benefits for metastatic renal cell carcinoma (mRCC), yet there is no current guidance regarding the optimum first-line regimen. We performed this network-meta analysis to compare the efficacy and safety of all available treatments for mRCC. Methods: A systematic search of literature was conducted in April 2019, and the analysis was done on a Bayesian fixed-effect model. Results: 25 randomized clinical trials (RCTs) involving 13,010 patients were included in this study. The results showed that for overall survival, pembrolizumab plus axitinib (hazard ratio [HR]: 0.53; 95% credible interval [CrI]: 0.38-0.73) and nivolumab plus ipilimumab (HR: 0.63; 95% CrI: 0.50-0.79) were significantly more effective than sunitinib, and pembrolizumab plus axitinib was probably (68%) to be the best choice. For progression-free survival, cabozantinib (HR: 0.66; 95% CrI: 0.46-0.94), pembrolizumab plus axitinib (HR: 0.69; 95% CrI: 0.57-0.84), avelumab plus axitinib (HR: 0.69; 95% CrI: 0.56-0.85), nivolumab plus ipilimumab (HR: 0.82; 95% CrI: 0.68-0.99), and atezolizumab plus bevacizumab (HR: 0.86; 95% CrI: 0.74-0.99) were statistically superior to sunitinib, and cabozantinib was likely (43%) to be the preferred options. Nivolumab plus ipilimumab (OR: 0.50; 95% CrI: 0.28-0.84), and atezolizumab plus bevacizumab (OR: 0.56; 95% CrI: 0.36-0.83) were associated with significantly lower rate of high-grade adverse events than sunitinib. Conclusions: Our findings demonstrate that pembrolizumab plus axitinib might be the best treatment for mRCC, while nivolumab plus ipilimumab has the most favorable balance between efficacy and acceptability, and may provide new guidance to make treatment decisions. Funding Statement: This research was supported by the Henan Provincial Scientific and Technological Research Project (Grant No. 192102310036). Declaration of Interests: The authors declare no conflicts of interest. Ethics Approval Statement: This study was performed based on the preferred reporting items for systematic reviews and meta-analyses (PRISMA) extension statement for network meta-analysis.

International audience; Trimethylamine N-oxide (TMAO) is widely dispersed in marine environments and plays an important role in the biogeochemical cycle of nitrogen. Diverse marine bacteria utilize TMAO as carbon and nitrogen sources or as electron acceptor in anaerobic respiration. Alteration of respiratory component according to the pressure is a common trait of deep-sea bacteria. Deep-sea bacteria from different genera harbor high hydrostatic pressure (HHP) inducible TMAO reductases that are assumed to be constitutively expressed in the deep-sea piezosphere and facilitating quick reaction to TMAO released from fish which is a potential nutrient for bacterial growth. However, whether deep-sea bacteria universally employ this strategy remains unknown. In this study, 237 bacterial strains affiliated to 23 genera of Proteobacteria, Bacteroidetes, Firmicutes and Actinobacteria were isolated from seawater, sediment or amphipods collected at different depths. The pressure tolerance and the utilization of TMAO were examined in 74 strains. The results demonstrated no apparent correlation between the depth where the bacteria inhabit and their pressure tolerance, regarding to our samples. Several deep-sea strains from the genera of Alteromonas, Halomonas, Marinobacter, Photobacterium, and Vibrio showed capacity of TMAO utilization, but none of the isolated Acinebacter, Bacillus, Brevundimonas, Muricauda, Novosphingobium, Rheinheimera, Sphingobium and Stenotrophomonas did, indicating the utilization of TMAO is a species-specific feature. Furthermore, we noticed that the ability of TMAO utilization varied among strains of the same species. TMAO has greater impact on the growth of deep-sea isolates of Vibrio neocaledonicus than shallow-water isolates. Taken together, the results describe for the first time the TMAO utilization in deep-sea bacterial strains, and expand our understanding of the physiological characteristic of marine bacteria.

Phosphorus (P), a significant biogenic element, plays a key role in the primary productivity of estuarine and coastal ecosystems. The study determined that the seasonal dynamics of total dissolved P (TDP)in the coastal area of the Zhanjiang Bay (ZJB) were largely affected by land-based sources. The TDP concentration had significant seasonal variations in the ZJB (P

In order to investigate the effect of hyperbranched polymer modified on impact strength of epoxy resin at 77K. One modifier called H204 was employed to methyl nadic anhydride (MNA) cured diglycidyl ether of bisphenol A (DGEBA) epoxy resin system. The impact strength at 77 K was studied and the results indicated that introduction of modifier used in this study can enhance the impact strength at cryogenic temperature. The 58.57% increase was obtained by addition of 10 wt% H204 into the epoxy matrix. Furthermore, scanning electron microscopy (SEM) was used to investigate the fracture mechanism and strengthening effect.

This paper presents an overview of research related to the role of fish farming on mercury (Hg) speciation and distribution in aquatic system. It critically reviews the major farm activities, e.g., organic waste loading, eutrophication and antibiotics using on the biogeochemical cycling of Hg. Hg is one of the most hazardous metals in the environment. The main exposure pathway of Hg to humans is through the consumption of fishery products. With the increasing consumption amount of fishery product, intensive fish farming activities increased dramatically and the production of aquaculture fish exceed the capture fish since 2013. However, intensive fish farming activities may change physical, chemical and biological characteristics of the benthic environment, which may have substantial influence on Hg speciation change and afterward bioaccumulation in fish through food chain. Thus, it is important to know how fish farming activities trigger Hg speciation change and distribution in the aquatic system, associated with the variation of Hg uptake by the aquaculture fish.

The preparation of sucrose fatty acid ester (SFAE) by lipase-catalyzing reaction usingCandida antarcticalipase B (CalB) and its immobilized form Novozym 435 was reported in this work. The preparation was characterized in non-aqueous media with and without ultrasound irradiation treatment. A conversion rate of SFAE up to 49.60% was achieved using Novozym 435 under the optimal conditions (45.4°C; mole ratio of methyl oleate to sucrose = 6.0:1; 4.0 mL acetone; 4.0 mg/mL Novozym 435; and 24.6 h of reaction). Under optimal ultrasound conditions (50 kHz, 0.15 W/cm2, 166.55 min), reaction time decreased by 75% approximately, compared with the control without ultrasonic irradiation, but the ultrasound irradiation treatment did not affect the SFAE yield catalyzed by Novozym 435. In the CalB-catalyzed preparation of SFAE under the same optimal reaction conditions, ultrasonic irradiation enhanced the activity of CalB during early time points and inhibited its activity after a long period of treatment. Moreover, CalB was further examined using Far-UV circular dichroism (CD) spectroscopy and scanning electron microscopy (SEM) to study the conformation and micro-morphology of CalB structural variations in various ultrasound irradiation treatments. CD results indicated that α-helical regions were increased and random coil regions remained at a similar level of proportion. SEM images showed small holes appeared on the surface of irradiated CalB. Therefore, we conclude that proper ultrasound irradiation could change the secondary structure and the surface morphology of the CalB in molecular level, and could accelerate the esterification reaction process.

A high performance liquid chromatography method using an efficient extraction method was developed for the determination of astaxanthin in eight kinds of animal feed. The chromatographic separation was achieved using a C18 reversed-phase column, using methanol and acetonitrile as the mobile phases with a flow rate of 1 mL/min. The feeds containing astaxanthin were first treated with maxatase, to cause enzymatic hydrolysis, and then extracted with dichloromethane. The optimized method produced recoveries of between 82.4% and 100% for all eight kinds of feed, and the coefficients of variation were lower than 4.28%. The limit of detection, defined as the concentration that gave a signal-to-noise ratio of 3, for astaxanthin was estimated to be 0.1 µg/g. The limit of quantification, defined as the lowest spiked concentration that gave an appropriate level of precision and accuracy, was 0.3 µg/g. Finally, the method developed was used to determine astaxanthin in real, commercially sourced, feed samples....

Quercetin (QE) and resveratrol (RSV) are powerful antioxidants with the potential to protect the testes against ischemia/reperfusion (I/R) injury. We compared their effects in testicular torsion/detorsion (T/D) in adult rats. Twenty-four male Wistar rats were divided into four groups: sham (group A), T/D (group B), T/D treated with QE (group C), and T/D treated with RSV (group D). QE (20 mg kg−1 ) and RSV (20 mg kg−1 ) were injected intra-peritoneally at 60 min of torsion. After 90 min of surgically induced torsion, the testicular cord was restored to its anatomical position. Twenty-four hour after torsion, blood and tissue samples were obtained for further examination. Testicular tissue malondialdehyde (MDA) and nitric oxide (NO) levels and serum total oxidant status (TOS) were higher in group B than in group A (P < 0.05). Group A had higher serum total antioxidant status (TAS) than group B. (P < 0.05) QE and RSV significantly lowered MDA, NO, and TOS levels and TAS consumption (P < 0.05). QE reduced the MDA and TOS levels more than RSV (P < 0.05), but their effects on NO reduction and TAS consumption were similar (P > 0.05). Group A had normal testicular architecture (grade 1). Groups C (mean grade 2.60) and D (mean grade 3.00) had lower testicular injury grades than group B (mean grade 3.45) (P < 0.05). Group C had lower testicular injury grade than group D (P < 0.05). Treatment with QE and RSV protects against I/R injury after testicular T/D. QE may exhibit better function than RSV at the doses tested in this study.

Herein, we developed a multi-analyte fluorescence immunoassay for detection of three organophosphate pesticides (triazophos, parathion, and chlorpyrifos) in various agro-products (rice, wheat, cucumber, cabbage, and apple) using fluorescently labeled oligonucleotide and gold nanoparticle (AuNP) signal amplification technology. The AuNP probes for the three analytes were constructed by simultaneously modifying the corresponding antibodies and fluorescently labeled oligonucleotides on the probe surface. Three fluorophores (6-FAM, Cy3, and Texas red) with high fluorescence intensity and little overlap of excitation/emission wavelengths were selected. The method showed satisfactory linearity for triazophos, parathion, and chlorpyrifos in the ranges of 0.01–20, 0.05–50, and 0.5–1000 μg/L, respectively. For the 3 analytes, the limits of detection (LODs) were 0.007, 0.009, and 0.087 μg/L, respectively. The average recoveries were 77.7–113.6%, with relative standard deviations (RSDs) of 7.1–17.1% in various food matrices. The proposed method offers great potential in food safety surveillance, and could be used as well as a reference for multi-residue analysis of other small-molecule contaminants.

PurposeThe optimum systemic therapies for advanced/metastatic renal cell carcinoma (RCC) of favourable, intermediate and poor risk have not been established. We aimed to compare and rank the effects associated with systemic therapies in the first-line setting.MethodsWe searched PubMed, Cochrane databases, Web of Science and ClinicalTrials.gov for randomised controlled trials (RCT) published up to February 2020 of all available treatments for advanced/metastatic RCC. Analysis was done on a Bayesian framework.Results15 unique RCTs including 8995 patients were identified. For advanced/metastatic RCC of favourable risk, avelumab plus axitinib was associated with a significantly higher improvement in progression-free survival (PFS) than sunitinib (HR 0.57, 95% CI 0.34 to 0.96). For intermediate-risk patients, cabozantinib, nivolumab plus ipilimumab, pembrolizumab plus axitinib and avelumab plus axitinib were associated with significantly higher improvement in PFS than sunitinib (HR 0.63, 95% CI 0.44 to 0.97; HR 0.66, 95% CI 0.53 to 0.81; HR 0.58, 95% CI 0.44 to 0.80; HR 0.62, 95% CI 0.47 to 0.83, respectively); pembrolizumab plus axitinib and nivolumab plus ipilimumab were associated with significantly higher improvement in overall survival (OS) than sunitinib (HR 0.53, 95% CI 0.34 to 0.81; HR 0.66, 95% CI 0.50 to 0.87, respectively). For poor-risk patients, nivolumab plus ipilimumab and pembrolizumab plus axitinib were associated with significantly higher improvement in PFS than sunitinib (HR 0.57, 95% CI 0.43 to 0.76; HR 0.48, 95% CI 0.30 to 0.82, respectively); nivolumab plus ipilimumab and pembrolizumab plus axitinib were significantly more efficacious for OS than sunitinib (HR 0.57, 95% CI 0.39 to 0.883; HR 0.43, 95% CI 0.23 to 0.80, respectively). For OS, there were 81% and 78% probabilities that pembrolizumab plus axitinib was the best option for intermediate-risk and poor-risk patients, respectively.ConclusionAvelumab plus axitinib might be the optimum treatment for advanced/metastatic RCC of favourable risk. Pembrolizumab plus axitinib might be the optimum treatment for intermediate-risk and poor-risk patients.

Phycocyanin, a type of functional food colorant, is shown to have a potent anti-cancer property. Non-small cell lung cancer (NSCLC) is one of the most aggressive form of cancers with few effective therapeutic options. Previous studies have demonstrated that phycocyanin exerts a growth inhibitory effect on NSCLC A549 cells. However, its biological function and underlying regulatory mechanism on other cells still remain unknown. Here, we investigated the in vitro function of phycocyanin on three typical NSCLC cell lines, NCI-H1299, NCI-H460, and LTEP-A2, for the first time. The results showed that phycocyanin could significantly induce apoptosis, cell cycle arrest, as well as suppress cell migration, proliferation, and the colony formation ability of NSCLC cells through regulating multiple key genes. Strikingly, phycocyanin was discovered to affect the cell phenotype through regulating the NF-κB signaling of NSCLC cells. Our findings demonstrated the anti-neoplastic function of phycocyanin and provided valuable information for the regulation of phycocyanin in NSCLC cells.

Tristyrylphenol ethoxylates (TSPxEOs), nonionic surfactant adjuvants, are widely used to replace alkylphenols in many agricultural formulations. Herein, a rapid and sensitive method based on a modified quick, easy, cheap, effective, rugged, and safe “QuEChERS” and supercritical fluid chromatography–tandem mass spectrometry (SFC–MS/MS) method was developed to quantify TSPxEOs in cucumber, leaves, and soil. Following optimization, the tested oligomers were fully separated within 8 min. The matrix-matched calibration standards showed satisfactory linearity with coefficients of determination (R2) ≥ 0.991. The limits of quantification (LOQs: 0.001–0.048 μg kg−1) were excellent for TSPxEOs (x = 6–29). The average recoveries at three (low, medium, and high) fortification levels were ranged between 62.5% and 112.9% with relative standard deviations (RSDs) ≤20.0% for TSPxEOs (x = 6–29) in various matrices. The method was successfully applied to determine TSPxEOs residual levels in field-incurred samples. Conclusively, the developed method is versatile and could be used for monitoring TSPxEOs in food and environmental samples.

We describe a single-step solvothermal method for the preparation of nanocomposites consisting of graphene oxide and Fe3O4 nanoparticles (GO/Fe3O4). This material is shown to be useful as a magnetic sorbent for the extraction of flavonoids from green tea, red wine, and urine samples. The nanocomposite is taking advantage of the high surface area of GO and the magnetic phase separation feature of the magnetic sorbent. The nanocomposite is recyclable and was applied to the extraction of flavonoids prior to their determination by HPLC. The effects of amount of surfactant, pH value of the sample solution, extraction time, and desorption condition on the extraction efficiency, and the regeneration conditions were optimized. The limits of detection for luteolin, quercetin and kaempferol range from 0.2 to 0.5 ng∙ mL−1 in urine, from 3.0 to 6.0 ng∙mL−1 in green tea, and from 1.0 to 2.5 ng∙mL−1 in red wine. The recoveries are between 82.0 and 101.4 %, with relative standard deviations of

To investigate human Hg exposure by food consumption and occupation exposure in a compact fluorescent lamp (CFL) manufacturing area, human hair and rice samples were collected from Gaohong town, Zhejiang Province, China. The mean values of total mercury (THg) and methylmercury (MeHg) concentrations in local cultivated rice samples were significantly higher than in commercial rice samples which indicated that CFL manufacturing activities resulted in Hg accumulation in local rice samples. For all of the study participants, significantly higher THg concentrations in human hair were observed in CFL workers compared with other residents. In comparison, MeHg concentrations in human hair of residents whose diet consisted of local cultivated rice were significantly higher than those who consumed commercial rice. These results demonstrated that CFL manufacturing activities resulted in THg accumulation in the hair of CFL workers. However, MeHg in hair were mainly affected by the sources of rice of the residents.

Porphyry deposits supply most of the world's Cu and Mo resources. Over 90% of the porphyry deposits are found at convergent margins, especially above active subduction zones, with much fewer occurrences at post-collisional or other tectonic settings. Porphyry Cu–(Mo)–(Au) deposits are essentially magmatic–hydrothermal systems, which are generally initiated by injection of oxidized magmas saturated with metal-rich aqueous fluids, i.e., the parental magmas need to be water rich and oxidized with most of the sulfur appearing as sulfate in the magma. Sulfur is the most important geosolvent that controls the behavior of Cu and other chalcophile elements, due to high partition coefficients of chalcophile elements between sulfide and silicate melts. Small amount of residual sulfides can hold a large amount of Cu. Therefore, it is essential to eliminate residual sulfides to get high Cu contents in magmas for the formation of porphyry deposits. Sulfate (SO 4 2 − ) is over 10 times more soluble than sulfide (S 2 − ), and thus the solubility of sulfur depends strongly on sulfur speciation, which in turn depends on oxygen fugacities. The magic number of oxygen fugacity is log f O 2 > FMQ + 2 (i.e., ΔFMQ + 2), where FMQ is the fayalite–magnetite–quartz oxygen buffer. Most of the sulfur in magmas is present as sulfate at oxygen fugacities higher than ΔFMQ + 2. Correspondingly, the solubility of sulfur increases from ~ 1000 ppm up to > 1 wt.%. Oxidation promotes the destruction of sulfides in the magma source, and thereby increases initial chalcophile element concentrations, forming sulfur-undersaturated magmas that can further assimilate sulfides during ascent. Copper, Mo and Au act as incompatible elements in sulfide undersaturated magmas, leading to high chalcophile element concentrations in evolved magmas. The final porphyry mineralization is controlled by sulfate reduction, which is usually initiated by magnetite crystallization, accompanied by decreasing pH and correspondingly increasing oxidation potential of sulfate. Hematite forms once sulfate reduction lowers the pH sufficiently, driving the oxidation potential of sulfate up to the hematite–magnetite oxygen fugacity (HM) buffer, which is ~ ΔFMQ + 4. Given that ferrous iron is the most important reductant that is responsible for sulfate reduction during porphyry mineralization, the highest oxygen fugacity favorable for porphyry mineralization is the HM buffer. In addition to the oxidation of ferrous iron during the crystallization of magnetite and hematite, reducing wallrock may also contribute to sulfate reduction and mineralization. Nevertheless, porphyry deposits are usually mineralized in the whole upper portion of the pluton, whereas interactions with country rocks are generally restricted at the interface, therefore assimilation of reducing sediments is not likely to be a decisive controlling process. Degassing of oxidized gases has also been proposed as a major process that is responsible for sulfate reduction. Degassing, however, is not likely to be a main process in porphyry mineralization that occurs at 2–4 km depths in the upper crust. Sulfide formed during sulfate reduction is efficiently scavenged by aqueous fluids, which transports metals to shallower depths, i.e., the top of the porphyry and superjacent wallrock. According to traditional views, sulfide saturation and segregation during magma evolution is not favorable for the formation of porphyry Cu ± Au ± Mo deposits. This is the main difference between porphyry deposits and Ni–Cu sulfide deposits. Nevertheless, in places with thick sections of reducing sediments, e.g., the western North America, sulfide saturation and segregation may occur during evolution of the magma, forming Cu-rich cumulates at the base of plutons. These Cu-rich sulfides may evolve into porphyry mineralization or even control the ore-forming process. Their contribution depends heavily on subsequent oxidation, i.e., a major contribution can be expected only when the sulfide cumulates are oxidized to sulfate, liberating the chalcophile elements. Sulfate reduction and ferrous Fe oxidation form H + , which dramatically lowers the pH values of ore-forming fluids and causes pervasive alteration zones in porphyry Cu deposits. The amount of H + released during mineralization and the alkali content in the porphyry together control the intensity of alterations. In principle, H 2 and methane form during the final mineralization process of porphyry deposits, but are mostly oxidized by ferric Fe during subsequent processes. Some of the reduced gases, however, may survive the highly oxidizing environment to escape from the system, or even to get trapped in fluid inclusions. Therefore, small amount of reduced gases in fluid inclusions cannot argue against the oxidized feature of the magmas. Reduced magmas are not favorable for porphyry mineralization. Reduced porphyry deposits so far reported are just mineralization that has either been reduced in host rock away from the causative porphyry or through assimilation of reducing components during emplacement.

An immunomagnetic-bead-based enzyme-linked immunosorbent assay (IMB-ELISA) was developed for detection of pesticides by using carboxyl functionalized magnetic Fe3O4 nanoparticles (CMNPs). The CMNPs were prepared by co-precipitation of Fe2+/Fe3+ with oleic acid as a surfactant and subsequent oxidation of CC into COOH by KMnO4 solution in situ. Then, anti-pesticide (triazophos) monoclonal antibodies were directly bonded onto the magnetic nanoparticles, which significantly increased the sensitivity compared with classic ELISA. The detection limit was 0.10 ng mL−1. Addition-recovery and high-precision experiments were performed on blank samples that were determined to be without triazophos. The average recovery rate for three types of samples (with each spiking concentration measured 5 times in parallel) ranged from 83.1% to 115.9%, with a relative standard deviation (RSD) of less than 10%, which meets the requirement of pesticide residue analysis. The application results were in accordance with the gas chromatography-mass spectrometry (GC-MS) method, suggesting that IMB-ELISA is rapid and reliable for pesticide detection.