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A study was conducted to investigate the inclusion effects of sugar beet pulp and rice straw mixture silage with inoculation (BRMS), in place of whole-plant corn silage (CS), on the dry matter intake, total-tract nutrient digestibility, plasma metabolites, rumen fermentation, and lactation performance in high-production dairy cows. Sixteen multiparous Holstein cows (body weight, 622 ± 35 kg; days in milk, 90 ± 11 d; mean ± standard deviation) were used in our experiments; the experiments were based on a repeated 4 × 4 Latin square design for 21 d, and each experimental period consisted of 14 d of adaptation, followed by 7 d of data collection. The 4 dietary treatments used were (dry matter basis): (1) 0% BRMS and 28.6% CS (0BRMS); (2) 4.3% BRMS and 24.3% CS (15BRMS); (3) 8.60% BRMS and 20.0% CS (30BRMS); and (4) 12.9% BRMS and 15.7% CS (45BRMS). The increasing inclusion of dietary BRMS was observed to linearly increase the total volatile fatty acids and the propionate concentration. The dry matter intake and digestibility values of neutral detergent fiber and acid detergent fiber increased linearly as the percentage of BRMS increased up to 45%. Milk yield linearly increased with the increase in the content of BRMS (39.0, 39.8, 40.9, and 40.3 kg/d for 0BRMS, 15BRMS, 30BRMS, and 45BRMS, respectively). The increasing inclusion of dietary BRMS induced a decrease in the ammonia nitrogen and milk urea nitrogen concentration, leading to a linear increase in milk protein production (1.15, 1.26, 1.35, and 1.27 kg/d for 0BRMS, 15BRMS, 30BRMS, and 45BRMS, respectively). In conclusion, the diets with the replacement of CS with BRMS up to 45% were beneficial to the production performance of high-production dairy cows, indicating that this method may be an appropriate use of sugar beet pulp and rice straw.

In this study, the impacts of transglutaminase (TGase)-induced glycosylation and limited enzymatic hydrolysis on the foaming property of soy protein isolates (SPI) and the possible underlying mechanisms were investigated. SPI were first glycosylated with oligochitosan by TGase to produce glycosylated samples (GSPI). GSPI were further hydrolyzed using Protamex with controlled degree of hydrolysis (DH of 1%, 2% and 4%). As compared to native SPI, the foaming stability of GSPI sample was increased from 24.3% ± 3.1% to 40.4% ± 7.6%, and the hydrolyzed GSPI showed enhanced foaming property. GSPI-1% DH showed the best foaming capacity and foam stability, increasing about 2.12 folds and 2.33 folds, respectively. Hydrodynamic-size of dominant distribution peak in GSPI and its hydrolysates (1% DH) were about 3090 and 531 nm, respectively, which were bigger than that of SPI (about 220 nm). The presence of oligochitosan in GSPI decreased surface hydrophobicity, and the subsequent hydrolysis generated an opposite effect. GSPI and its hydrolysates had more flexible tertiary conformations as revealed by intrinsic fluorescence study. Overall, high values of negative zeta potential and apparent viscosity of samples might be responsible for the improved foaming property. This study suggested that TGase-induced glycosylation and limited enzymatic hydrolysis are promising techniques for enhancing the foaming property of SPI, which would broaden the applications of SPI in food industry.

Baeyer–Villiger (BV) oxidation is a fundamental organic reaction, whereas the hetero-BV oxidation is uncharted. Herein, a tellura-BV oxidation is discovered. By oxidizing a tellurophene-embedded and electron-rich polycycle (1) with mCPBA or Oxone, an oxygen atom is inserted into the Te–C bond of the tellurophene to form tellurinate lactone mono-2. This reaction proceeds as follows: (i) 1 is oxidized to the tellurophene Te-oxide form (IM-1); (ii) IM-1 undergoes tellura-BV oxidation to give mono-2. Moreover, the hybrid trichalcogenasumanenes 7 and 8 are, respectively, converted to tellurinate lactones mono-9 and mono-10 under the same conditions, indicating that tellura-BV oxidation shows high chemoselectivity. Due to the strong secondary bonding interactions between the Te Created by potrace 1.16, written by Peter Selinger 2001-2019 O groups on tellurinate lactones, mono-2, mono-9, and mono-10 are dimerized to form U-shaped polycycles 2, 9, and 10, respectively. Notably, mono-2, mono-9, mono-10, and their dimers show chirality. This work enables one-step transformation of tellurophene into tellurinate lactone and construction of intricate polycycles., A tellura-Baeyer–Villiger oxidation is discovered, which enables one-step transformation of tellurophene into chiral tellurinate lactones. The resulting tellurinate lactones are dimerized in the solid state to form U-shaped polycycles.

A novel fibrinolytic enzyme, ACase was isolated from fruiting bodies of a mushroom, Agrocybe aegerita. ACase was purified by using ammonium sulfate precipitation, gel filtration, ion exchange and hydrophobic chromatographies to 237.12 fold with a specific activity of 1716.77 U/mg. ACase was found to be a heterodimer with molecular mass of 31.4 and 21.2 kDa by SDS-PAGE and appeared as a single band on Native-PAGE and fibrin-zymogram. The N-terminal sequence of the two subunits of ACase was AIVTQTNAPWGL (subunit 1) and SNADGNGHGTHV (subunit 2). ACase had maximal activity at 47 °C and pH 7.6. It's activity was improved by Cu2+, Na+, Fe3+, Zn2+, Ba2+, K+ and Mn2+, but inhibited by Fe2+, Mg2+ and Ca2+. PMSF, SBTI, aprotinine and Lys inhibited the enzyme activity, which suggested that ACase was a serine protease. ACase could degrade all three chains (α, β and γ) of fibrinogen. Moreover, the enzyme acted as both, a plasmin-like fibrinolytic enzyme and a plasminogen activator. It could hydrolyze human thrombin slightly, which indicated that the ACase could inhibit the activity of thrombin and acted as an anticoagulant to prevent thrombosis. Based on these results, ACase might act as a therapeutic agent for treating thrombosis, or as a functional food. Further investigation of the enzyme is underway.

In this study, magnetic Fe3O4@SiO2@β-cyclodextrin copolymerized microparticles were synthesized and applied for the extraction of methyl parathion and fenthion in lettuce samples followed by HPLC-UV detection. The magnetic β-cyclodextrin copolymerized microparticles were prepared by dispersion polymerization with acryloyl β-cyclodextrin as the functional monomer and ethylene glycol dimethyacrylate as the crosslinker. The composite magnetic microparticles were characterized by Fourier transform infrared spectroscopy, X-ray diffraction, magnetic measurement, and thermogravimetric analysis, and used as the adsorbent of magnetic solid-phase extraction (MSPE) for methyl parathion and fenthion. The extraction conditions including sample pH and ionic strength, desorption solvent type and volume, and adsorption and desorption times were optimized. Under the optimal extraction conditions, an MSPE-HPLC-UV method was developed for the detection of methyl parathion and fenthion in lettuce. Wide linear ranges of 1.0-200 μg kg-1 (R2 = 0.9998) for methyl parathion and 1.5-200 μg kg-1 (R2 = 0.9978) for fenthion were obtained and the limits of detection were 0.3 μg kg-1 for methyl parathion and 0.5 μg kg-1 for fenthion in lettuce, respectively. The proposed method was applied for the determination of methyl parathion and fenthion in lettuce with satisfactory recoveries between 89.2-101.2%, and relative standard deviations were less than 9.1%. Thus, the MSPE-HPLC-UV method has high accuracy and sensitivity for the analysis of methyl parathion and fenthion in lettuce samples.

Objective Arteriosclerosis is an age-related disease and a leading cause of cardiovascular disease. In animal experiments, mesenchymal stem cells and its culture-conditioned medium have been shown to be promising tools for prevention or treatment of arteriosclerosis. On the basis of these evidences, we aimed to assess whether administration of autologous adipose-derived mesenchymal stem cells (Ad-MSC) is safe and effective for treatment of arteriosclerosis. Methods We retrospectively reviewed clinical records of patients with arteriosclerosis who had received autologous Ad-MSC administration at our clinic. Patients’ characteristics were recorded and data on lipid profile, intimal-media thickness (IMT), cardio-ankle vascular index (CAVI), and ankle-brachial index (ABI) before and after Ad-MSC administration were collected and compared. Results Treatment with Ad-MSC significantly improved HDL, LDL, and remnant-like particle (RLP) cholesterol levels. No adverse effect or toxicity was observed in relation to the treatment. Of the patients with abnormal HDL values before treatment, the vast majority showed improvement in the values. Overall, the measurements after treatment were significantly increased compared with those before treatment (p p Conclusions These findings suggest that Ad-MSC administration is safe and effective in patients developing arteriosclerosis, thereby providing an attractive tool for anti-aging application.

Approximately 50-80% of the world population are infected with H. pylori, which is categorized as a class I carcinogen. Antiadhesive therapy is emerging as a promising alternative to antibiotics against bacterial infection. This study demonstrated that defatted wheat germ protein hydrolysates (DWGPH) effectively inhibited H. pylori adhesion to gastric epithelial cells. DWGPH prepared by pronase possessed the best activity where its inhibitory percentage at 10 mg/mL was 51.7 ± 6.8% and the minimum antiadhesive concentration was 0.31 mg/mL. The antiadhesive activity is attributable to peptides acting as receptor analogs in binding to H. pylori. Peptides with potential H. pylori-binding ability (n = 267) were identified, and their structural characteristics were comprehensively analyzed, including net charge, Boman index, instability index, aliphatic index, molecular weight, isoelectric point, hydrophobicity, and Hmoment (α-helix and β-sheet). This work provided an array of peptide sequences for further exploration as putative ligands of H. pylori adhesins and for elucidating molecular mechanisms.

NOX5 (NADPH oxidase 5) is a homolog of the gp91 phox subunit of the phagocyte NOX, which generates reactive oxygen species. NOX5 is involved in sperm motility and vascular contraction and has been implicated in diabetic nephropathy, atherosclerosis, and stroke. The function of NOX5 in the cardiac hypertrophy is unknown. Because NOX5 is a Ca 2+ -sensitive, procontractile NOX isoform, we questioned whether it plays a role in cardiac hypertrophy. Studies were performed in (1) cardiac tissue from patients undergoing heart transplant for cardiomyopathy and heart failure, (2) NOX5-expressing rat cardiomyocytes, and (3) mice expressing human NOX5 in a cardiomyocyte-specific manner. Cardiac hypertrophy was induced in mice by transverse aorta coarctation and Ang II (angiotensin II) infusion. NOX5 expression was increased in human failing hearts. Rat cardiomyocytes infected with adenoviral vector encoding human NOX5 cDNA exhibited elevated reactive oxygen species levels with significant enlargement and associated increased expression of ANP (atrial natriuretic peptides) and β-MHC (β-myosin heavy chain) and prohypertrophic genes ( Nppa , Nppb , and Myh7 ) under Ang II stimulation. These effects were reduced by N-acetylcysteine and diltiazem. Pressure overload and Ang II infusion induced left ventricular hypertrophy, interstitial fibrosis, and contractile dysfunction, responses that were exaggerated in cardiac-specific NOX5 trangenic mice. These phenomena were associated with increased reactive oxygen species levels and activation of redox-sensitive MAPK (mitogen-activated protein kinase). N-acetylcysteine treatment reduced cardiac oxidative stress and attenuated cardiac hypertrophy in NOX5 trangenic. Our study defines Ca 2+ -regulated NOX5 as an important NOX isoform involved in oxidative stress- and MAPK-mediated cardiac hypertrophy and contractile dysfunction.

Increased use of nitrogen fertilizers has deleterious impact on the environment. Increase in yield potential at low nitrogen supply is regarded as a cereal breeding goal for future agricultural sustainability. Although natural variations of nitrogen transporters have been investigated, key genes associated with assimilation remain largely unexplored for nitrogen use efficiency (NUE) enhancement. Here, we identified a NIN‐like protein NLP4 associated with NUE through a GWAS in rice. We found that OsNLP4 transactivated OsNiR encoding nitrite reductase that was critical in nitrogen assimilation in rice. We further constructed quadrupling NREs (Nitrate‐responsive cis‐elements) in the promoter of OsNiR (p4xNRE:OsNiR) and enhanced nitrogen assimilation significantly. We demonstrated that OsNLP4‐OsNiR increased tiller number and yield through enhancing nitrogen assimilation and NUE. Our discovery highlights the genetic modulation of OsNLP4‐OsNiR signalling cascade as a strategy for high NUE and yield breeding in rice.

In mass casualty events involving radiation exposure, there is a substantial unmet need for identifying and developing an orally bioavailable agent that can be used to protect the hematopoietic stem cell pool and regenerate hematopoiesis after radiation injury. Dimethyl sulfoxide (DMSO), a free-radical scavenger, has shown therapeutic benefits in many preclinical and clinical studies. This study investigates the radioprotective effects of DMSO on oral administration. Single dose of oral DMSO administrated before irradiation conferred 100% survival of C57BL6/J mice receiving otherwise lethal as well as super-lethal radiation dose, with wide radioprotective time frame (from 15min to 4h). Oral DMSO not only protected radiation-induced acute hematopoietic stem and progenitor cell (HSPC) injury, but also ameliorated long-term BM suppression following irradiation in mice. Mechanistically, DMSO directly protected HSPC survival after irradiation in vitro and in vivo, whereas no radioprotective effect was seen in MLL-AF9-induced leukemia cells. Unexpectedly, DMSO treatment did not inhibit radiation-induced HSPC apoptosis, and the HSPC survival from Trp53-and PUMA-deficient mice after irradiation was also protected by DMSO. In conclusion, our findings demonstrate the radioprotective efficacy of oral DMSO. Given its oral efficacy and little toxicity, DMSO is an attractive candidate for human use in a wide variety of settings, including nuclear accidents and medical radiation.

In this study, porous sandwich structure Fe3 O4 nanoparticles coated by polyhedral oligomeric silsesquioxanes and β-cyclodextrin were prepared by surface polymerization and were used as the magnetic solid phase extraction adsorbent for the extraction and determination of carbaryl and carbofuran. The Fe3 O4 nanoparticles coated with polyhedral oligomeric silsesquioxanes and β-cyclodextrin were characterized by Fourier transform infrared spectroscopy, X-ray diffraction, thermogravimetric analysis, vibrating sample magnetometry, and scanning electron microscopy. After optimizing the extraction conditions, a method that combined magnetic solid phase extraction with high-performance liquid chromatography was developed for the determination of carbaryl and carbofuran in apple. The method exhibited a good linearity in the range of 2-400 μg/kg for carbaryl and carbofuran (R2 = 0.9995), respectively. The limits of detection were 0.5 μg/kg of carbaryl and 0.7 μg/kg for carbofuran in apple, respectively. Extraction recoveries ranged from 94.2 to 103.1% with the preconcentration factor of 300 and the relative standard deviations were less than 5.9%. These results indicated that the method combined magnetic solid phase extraction with high-performance liquid chromatography and was promising for the determination of carbaryl and carbofuran at trace amounts.

Alcoholic liver disease is one of the most prevalent chronic diseases that causes high morbidity and mortality worldwide. This study demonstrated the protective potential of Alcalase-black soybean-derived peptides (BSPs) against alcohol-induced liver injury in mice. BSPs were rich in oligopeptides (80%, w/w, 0.05). All treatments significantly (P

Fibrinolytic enzymes are important thrombolytic agents for blood-clotting disorders like cardiovascular diseases. Availability of novel recombinant fibrinolytic enzymes can overcome the shortcomings of current thrombolytic drugs. With the objective of facilitating their cost-effective production for therapeutic applications and for gaining deeper insight into their structure-function, we have cloned and expressed the first fibrinolytic protease gene from Cordyceps militaris. Cordyceps militaris fibrinolytic enzyme (CmFE) has one open reading frame of 759 bp encoding “pre-pro-protein” of 252 amino acids. Recombinant CmFE was expressed as 28 kDa extracellular enzyme in Pichia pastoris which was capable of degrading fibrin clot. A structure homology model of CmFE was developed using urokinase-type plasminogen activator. The active site contains catalytic triad His41, Asp83, Ser177 and consensus sequence of GDSGG. The substrate binding residues are Asp (171), Gly (194) and Ser (192). Its trypsin-like specificity is determined by the critical Asp171 in S1 subsite. The “oxyanion hole” is formed by backbone amide hydrogen atoms of Gly-175 and Ser-177. CmFE contains six conserved cysteines forming three disulfide linkages. This is the first study describing cloning, expression and prediction of structure-function relationship of a mushroom fibrinolytic protease. Hence it has great relevance in application of fibrinolytic enzymes as thrombolytic agents.

Novel mesoporous yolk–shell structure magnetic molecularly imprinted polymers (MYS-MMIPs) were synthesized via surface polymerization using E2 as a template followed by chemical selective etching to remove the inorganic silica interlayer for the rapid, selective extraction and sensitive detection of 17β-estradiol (E2) in milk powder. For MYS-MMIPs, transmission electron microscopy coupled with N2 adsorption experiments confirmed the successful formation of a mesoporous yolk–shell structure. Furthermore, the thermogravimetric and recycling experiments showed the good thermal and adsorption stability of MYS-MMIPs, the magnetic properties experiment confirmed their good magnetic response, and the binding experiment indicated their selective recognition and high binding capacity (39.24 mg g−1) for E2. The obtained polymers were applied as adsorbents for the magnetic solid phase extraction (MSPE) of E2. After optimizing the MSPE conditions, an MSPE-HPLC method was developed for the determination of E2 in milk powder. The linear range was 1–300 μg kg−1 with a limit of detection of 0.33 μg kg−1. The satisfactory recoveries ranged from 88.3–102.4% and the relative standard deviations (RSDs) were lower than 5.5%. Thus, the MSPE-HPLC method has high accuracy and sensitivity for the analysis of E2 in milk samples.

Over-application of nitrogen fertilizer in fields has had a negative impact on both environment and human health. Domesticated rice varieties with high nitrogen use efficiency (NUE) reduce fertilizer for sustainable agriculture. Here, we perform genome-wide association analysis of a diverse rice population displaying extreme nitrogen-related phenotypes over three successive years in the field, and identify an elite haplotype of nitrate transporter OsNPF6.1HapB that enhances nitrate uptake and confers high NUE by increasing yield under low nitrogen supply. OsNPF6.1HapB differs in both the protein and promoter element with natural variations, which are differentially trans-activated by OsNAC42, a NUE-related transcription factor. The rare natural allele OsNPF6.1HapB, derived from variation in wild rice and selected for enhancing both NUE and yield, has been lost in 90.3% of rice varieties due to the increased application of fertilizer. Our discovery highlights this NAC42-NPF6.1 signaling cascade as a strategy for high NUE and yield breeding in rice., Improving crop nitrogen use efficiency can facilitate sustainable production, however, the genetic mechanisms have not been fully revealed. Here, the authors discover the NAC42-NPF6.1 signaling cascade mainly derives from indica and wild rice and demonstrate the potential of using the allele for cultivar improvement.

Aluminum antimonide (AlSb) is a promising semiconductor candidate for photovoltaic and opto-electronic applications. In this study, ∼200 μm thick AlSb films were prepared by screen printing and sintering method. To clarify the phase formation process during sintering, TG/DSC analysis, XRD analysis, SEM observations (combined with EDS line scans) and DC conductivity measurements were carried out. DSC curves and XRD patterns confirmed that the solid–solid reaction, Sb melting, solid–liquid reaction, Al melting, liquid–liquid reaction occurred in sequence with increasing temperature within the Al-Sb system, and AlSb phase was mainly generated at the liquid–liquid reaction stage. Moreover, SEM characterizations illustrated that the films sintered at liquid-liquid stage showed extremely different morphologies with the films sintered at solid–solid and solid–liquid reaction stages, the films were highly developed and large AlSb particles distributed in them. Finally, the semiconducting nature of the films was affirmed by DC conductivity measurements, both resistivity and activation energy increased with increasing sintering temperature owing to the formation of more AlSb phase under higher sintering temperatures.

Increasing use of TiO2 nanoparticles (NPs) leads to an accumulation in environment and a risk to human. In this work, an on-line monolithic capillary microextraction (CME)-inductively coupled plasma mass spectrometry (ICP-MS) method was developed for sensitive determination of TiO2 NPs in environmental water samples. With the presence of citric acid, TiO2 NPs can be selectively adsorbed on the poly(acrylamide)-vinylpyridine-methylene bis-(acrylamide) (poly(AA-VP-Bis)) monolithic column at pH 1, whereas Ti ions cannot be adsorbed. Factors affecting the extraction efficiency such as sample pH, concentration of citric acid and elution conditions were carefully investigated. Under the optimum conditions, a detection limit of 0.63 μg L−1 for TiO2 NPs and a sample throughput of 6 h−1 were achieved. The method can be applied to the determination of TiO2 NPs in the size range of 5–100 nm. The developed method was then applied to the analysis of TiO2 NPs in the East Lake water and Yangtze River water, and recoveries of 84–109% were obtained for spiked samples. Compared with other reported methods for the determination of metal oxide NPs, the developed on-line monolithic CME-ICP-MS has the merits of simple operation, high throughput and no need for digestion before ICP-MS detection.

Temperature compensation is a fundamental property of all circadian clocks; temperature compensation results in a relatively constant period length at different physiological temperatures, but its mechanism is unclear. Formation of a stable complex between clock proteins and casein kinase 1 (CK1) is a conserved feature in eukaryotic circadian mechanisms. Here, we show that the FRQ-CK1 interaction and CK1-mediated FRQ phosphorylation, not FRQ stability, are main mechanisms responsible for the circadian temperature compensation phenotypes in Neurospora. Inhibition of CK1 kinase activity impaired the temperature compensation profile. Importantly, both the loss of temperature compensation and temperature overcompensation phenotypes of the wild-type and different clock mutant strains can be explained by temperature-dependent alterations of the FRQ-CK1 interaction. Furthermore, mutations that were designed to specifically affect the FRQ-CK1 interaction resulted in impaired temperature compensation of the clock. Together, these results reveal the temperature-compensated FRQ-CK1 interaction, which results in temperature-compensated CK1-mediated FRQ and WC phosphorylation, as a main biochemical process that underlies the mechanism of circadian temperature compensation in Neurospora. IMPORTANCE Temperature compensation allows clocks to adapt to all seasons by having a relatively constant period length at different physiological temperatures, but the mechanism of temperature compensation is unclear. Stability of clock proteins was previously proposed to be a major factor that regulated temperature compensation. In this study, we showed that the interaction between CK1 and FRQ, but not FRQ stability, explains the circadian temperature compensation phenotypes in Neurospora. This study uncovered the key biochemical mechanism responsible for temperature compensation of the circadian clock and further established the mechanism for period length determination in Neurospora. Because the regulation of circadian clock proteins by CK1 and the formation of a stable clock complex with CK1 are highly conserved in eukaryotic clocks, a similar mechanism may also exist in animal clocks.

Cancer expression of PD-L1 suppresses anti-tumor immunity. PD-L1 has emerged as a remarkable therapeutic target. However, the regulation of PD-L1 degradation is not understood. Here, we identify several compounds as inducers of PD-L1 degradation using a high-throughput drug screen. We find EGFR inhibitors promote PD-L1 ubiquitination and proteasomal degradation following GSK3α-mediated phosphorylation of Ser279/Ser283. We identify ARIH1 as the E3 ubiquitin ligase responsible for targeting PD-L1 to degradation. Overexpression of ARIH1 suppresses tumor growth and promotes cytotoxic T cell activation in wild-type, but not in immunocompromised mice, highlighting the role of ARIH1 in anti-tumor immunity. Moreover, combining EGFR inhibitor ES-072 with anti-CTLA4 immunotherapy results in an additive effect on both tumor growth and cytotoxic T cell activation. Our results delineate a mechanism of PD-L1 degradation and cancer escape from immunity via EGFR-GSK3α-ARIH1 signaling and suggest GSK3α and ARIH1 might be potential drug targets to boost anti-tumor immunity and enhance immunotherapies., The regulation of PD-L1 via proteasomal degradation is unclear. Here, the authors show that EGFR inhibition activates GSK3 α to promote PD-L1 phosphorylation, which leads to PD-L1 ubiquitination and proteasome mediated degradation by ARIH1 E3 ligase.

Background: The immunotherapy for different types of cancers that targeting programmed death protein-1 (PD-1) and programmed death ligand-1 (PD-L1) has highlighted the importance of suppressing specific T cell responses. Recently, several studies have shown that the expression level of PD-L1 in tumor cells is positively correlated with tumor metastasis as well as recurrence rate. The potent effects of post-translational modifications (PTMs) for PD-L1, such as ubiquitination, glycosylation, phosphorylation and palmitoylation, have been reported to be related to immunosuppression. However, the regulation of PD-L1 degradation in cancers is still not well understood. In this paper, we mainly investigate the deubiquitination regulation of PD-L1. Methods: The protein levels of PD-L1 and USP21 were detected by Immunoblotting and immunohistochemistry. The interaction between PD-L1 and USP21 was determined by co-immunoprecipitation. The deubiquitination of PD-L1 was determined by in vitro deubiquitination assay. The deubiquitination sites of PD-L1 were identified by mass spectrometry analysis. The expression of mRNA in target tissues was presented by bioinformatics analysis.Results: Overexpression of USP21 significantly increased PD-L1 abundance and knockdown of USP21 induced degradation of PD-L1. In vitro deubiquitination assay showed that USP21-WT reduced polyubiquitin chains from PD-L1 while USP21-C221A did not. Furthermore, five lysines in intracellular segment of PD-L1 are potential deubiquitin sites and cancer-derived mutations of PD-L1 in Asp276 have the ability to enhance the deubiquitination of PD-L1 mediated by USP21. Finally, we found that USP21 is the frequently amplified deubiquitinase in lung cancer, especially in lung squamous cell carcinoma, and its amplification co-occurs with the upregulation of PD-L1 levels. Moreover, IHC analysis showed stronger staining of PD-L1 and USP21 in lung cancer samples than adjacent tissues. Conclusion: We identified USP21 as a novel deubiquitinase of PD-L1. Hopefully, targeting PD-L1 by inhibiting USP21 might be a potentially novel strategy for the treatment of lung cancer.

Over the last decade, the controllable reversible phase transition of functional materials has received growing interest as it shows unique suitability for various technological applications. Although many metal-organic frameworks (MOFs) possess a lamellar structure, the reversible structural transformation of MOFs between their three-dimensional (3D) phase and two-dimensional (2D) phase remains a largely unexplored area. Herein, we report for the first time a europium MOF with unprecedented reversible morphology in different solvents at room temperature. This europium MOF displayed a 3D nanorod morphology in organic solvent and a 2D nanobelt architecture in water. As a proof of concept for potential applications of this reversible-phase-transition MOF, we were able to use a delamination recovery method to load dye molecules that previously could not be loaded into europium MOFs.

To improve the electrochemical performance of CFx for lithium primary batteries, a ball milling treatment of CFx and urea with various mass ratios is carried out in this paper. It is shows that the weight ratio of urea/CFx significantly affects the electrochemical performance of the ball-milled CFx. Electrochemical tests show that the modified CFx material exhibits excellent electrochemical performance with much enhanced rate capacity, improved discharge platform, and lowered initial potential delay compared with the untreated CFx. In detail, when the weight ratio of urea/CFx is 5, the material delivers a discharge capacity of 550.6 mAh g−1 with a high power density of 10309 W kg−1 (at 5000 mA g−1 and 25 °C) and exhibits an excellent electrochemical stability at low temperature of 5 °C (power density up to 5922.5 W kg−1 at 3000 mA g−1). The outstanding electrochemical performance is mainly due to the synergistic effect of enlarged interlayer distance, decreased particle size, and increased surface area, which resulting in improved the electrochemical reaction activity, decreased reaction resistance, and facilitated lithium ions diffusion.

New glycopeptides were generated by proteolysis from corn gluten meal (CGM) followed by transglutaminase (TGase)-induced glycosylation with glucosamine (GlcN). The glycopeptides exhibited desirable antioxidant and intracellular ROS-scavenging properties. The amount of conjugated GlcN quantified by high-performance liquid chromatography (HPLC) was 23.0 g/kg protein. The formed glycopeptides contained both glycosylated and glycation types, as demonstrated by the electrospray ionization time-of-flight mass spectrometry (ESI-TOF MS/MS). The glycopeptides exhibited scavenging capabilities against free radical diphenylpicrylhydrazyl (DPPH) and hydroxyl radicals by reducing their power. The potential protection of glycopeptides against ethanol-induced injury in LO2 cells was assessed In Vitro based on methyl thiazole tetrazolium (MTT) testing and intracellular reactive oxygen species (ROS) scavenging capacity, respectively. Glycopeptide cytoprotection was expressed in a dose-dependent manner, with the glycopeptides exhibiting good solubility ranging from 74.8% to 83.2% throughout a pH range of 2&ndash, 10. Correspondingly, the glycopeptides showed good emulsifying activity (36.0 m2/g protein), emulsion stability (74.9%), and low surface hydrophobicity (16.3). These results indicate that glycosylation of CGM significantly improved its biological and functional properties. Glycopeptides from CGM could be used as potential antioxidants as well as comprising a functional food ingredient.

Non-toxic and long-term fluorescent probes for tumor imaging are in urgent need for non-invasively obtaining information about tumor genesis and metastasis in vivo. Here, we present a biocompatible near-infrared fluorescent probe for in vivo long-term imaging of tumor by modifying glucose (Glc), which experiences high uptake in cancer cells, on the surface of near-infrared Ag2Se quantum dots (NIR Ag2Se QDs). The fluorescence of glucose-functionalized Ag2Se QDs (Glc-Ag2Se QDs) from the targeted tumor can be observed in vivo for at least 7 days. In addition, this probe could be excreted through kidneys and the renal excretion ability is favorable for in vivo imaging applications. Moreover, Glc-Ag2Se QDs could be used for tumor targeted imaging of not only human breast cancer cells (MCF-7), but also SW1990 pancreatic cancer cells since glucose is highly taken up in almost all kinds of tumors. Glc-Ag2Se QDs could be a promising general tool for in vivo long-term observation of tumor evolution.

The intermolecular interactions and molecular packing form of fluorescent molecules have a huge impact on their optical properties, especially for AIE molecules. As a class of typical AIE molecules, tetraphenylethene (TPE) and its derivatives have prominent optical properties; nevertheless, separation of mixtures of E/Z isomers is a great challenge. Herein, a series of aromatic-substituted TPE derivatives were synthesized and used to separate mixtures of E/Z isomers by common column chromatography with high yields, as confirmed by single crystal analysis, mass spectrometry and NMR spectroscopy. The structure–property relationships of these molecules were systematically investigated by a combination of spectroscopic methods, theoretical calculations and single crystal data analysis. E/Z isomers exhibit many different fluorescent properties, such as AIE and mechanochromic behavior. Moreover, the position of N on the substituted pyridine ring also has an effect on the molecule stack pattern and the fluorescent properties. Collectively, our findings could not only improve the fundamental understanding of the cis/trans isomerization and photophysical properties of TPE derivatives but also provide a good strategy for designing different substituted groups that can produce various functions and have more potential applications.

Covalent organic frameworks (COFs) are a class of crystalline porous materials utilized in various potential applications. However, the development of hollow-structured COFs with defined morphology is important for their further applications, which is rare. Herein, COF with unique hollow shuttle morphology was prepared by a Schiff condensation reaction between 4-(4-aldehyde phenyl)ethylene (TPE) and tetra-(4-aminophenyl) porphyrin (TAP). A detailed mechanistic investigation reveals that an initial self-assembly followed by a similar inside-out Ostwald ripening process is responsible for the hollow capsule formation. The hollow microshuttle-shaped capsule COF is used for studying hemoglobin adsorption, which shows an uptake of 550.82 mg g-1 of hemoglobin. These studies could foreshadow new avenues for the development of porous materials with defined morphologies for the adsorption of biomolecules.

Here, a new 1D helical chain coordination polymer with enhanced chemical stability and fluorescence emission was prepared by introducing a benzimidazole molecule into Ag12 clusters. This coordination polymer demonstrated an increased fluorescence intensity with increasing polarity of alcohol solution, revealing its potential application in alcohol sensing.

Enzyme-mimicking nanomaterials (nanozymes) have been used to catalyze enzyme-like reactions for various applications. Despite these favorable properties, peroxidase-mimicking nanozymes suffer from the optimum reaction occurring in acidic solutions, which greatly limits their applications in biological systems. Herein, we demonstrate that copper Metal–Organic Polyhedra (Cu-MOP) nanorods exhibit high peroxidase-mimicking activity at a physiological pH value. Cu-MOP nanorods catalyze the generation of strong chemiluminescence signals in the presence of luminol/H2O2 and the oxidation of 2,2′-azinobis-(3-ethylbenzthiazoline-6-sulphonate acid) (ABTS) by H2O2 produces a green color reaction in a neutral pH solution. The Cu-MOP nanorod driven generation of color reactions is applied to develop a H2O2 sensor, and furthermore is implemented to develop a glucose detection platform.

To develop a convenient method for sensitive and selective determination of Ce3+ in aqueous phase with complicated matrices, a carbon paste electrode (CPE) modified with ion imprinted polymers (IIPs) were fabricated. The polymers were prepared by precipitation polymerization using Ce3+ as template, allyl phenoxyacetate (APA) as monomer, ethylene glycol dimethacrylate (EGDMA) as crosslinker and azobisisobutyronitrile (AIBN) as initiator under the molar ratio of Ce3+, APA and EGDMA as 1:4:40, respectively. Ce3+ was detected directly by differential pulse adsorptive stripping voltammetry (DPASV) and its oxidation peak appears at about 0.93 V. All parameters affecting the sensor's response are optimized and a calibration curve is plotted at a linear range of 1.0 × 10−6–1.0 × 10−5 mol/L and 1.0 × 10−5–2.0 × 10−4 mol/L with the detection limit of 1.5 × 10−7 mol/L. All other rare earth ions have no interference with the determination of Ce3+ even at a concentration 500 times higher than that of Ce3+. This sensor was successfully applied to determination of Ce3+ in two catalyst sample solutions with RSD≤3.3% (n = 5) and recoveries in the range of 99.2%–106.5% at our optimal conditions.

In this study, we have isolated and characterized a fibrinolytic enzyme from the GRAS (Generally Recognized as Safe) fungus, Neurospora sitophila. The enzyme was purified by fractional ammonium sulfate precipitation, hydrophobic interaction, ion exchange and gel filtration chromatography to 45.2 fold with a specific activity of 415.6U/mg protein. The native molecular mass of the enzyme was 49kDa, while the denatured molecular mass was 30kDa and 17.5kDa, indicating that the enzyme was a hetero-dimer. It was optimally active at 50°C and pH 7.4 and stable at human physiological temperature and pH. It was found to be a chymotrypsin-like serine protease which cleaved the synthetic chromogenic substrate, N-Succinyl-Ala-Ala-Pro-Phe-pNA for which the apparent Km and Vmax values were 0.24mM and 4.17×10-5mM/s, respectively. The enzyme hydrolyzed all the chains of fibrinogen by cleaving α chain first, followed by β chain and then γ chain. Moreover, the enzyme possessed dual function of direct fibrinolysis as well as plasminogen activation. Due to its attractive biochemical and fibrinolytic properties and being from a GRAS fungus, the fibrinolytic enzyme has application as a safe and efficient thrombolytic drug.

Zein and its derived peptides have been used as nanocarriers for bioactive components. Lutein, as well as other xanthophylls, are characterized by blue light filtering and anti-oxidant properties. However, lutein is unstable and has low water solubility, poor absorption, and low bioavailability. In order to protect lutein from oxidative degradation, and to enhance its solubility and dispersibility, stability and bioactivity, lutein-loaded zein nanoparticles (LLZ-NP) and zein-derived peptide nanoparticles (LLZ-PEP-NP) were prepared by the solvent diffusion method. Compared to LLZ-NP, LLZ-PEP-NP possessed good physicochemical properties, including particle size, polydispersity index, zeta potential, entrapment efficiency and in vitro stability. Specifically, transmission electron microscopy (TEM) images showed that LLZ-PEP-NP had a spherical form with a nanometric size and lutein was efficiently loaded into zein-derived peptides through self-assembly. Dynamic light scattering (DLS) results demonstrated that LLZ-PEP-NP had a narrow size distribution in the range of 200-300 nm and a decreased zeta potential compared to that of LLZ-NP. The lutein entrapment efficiency (EE%) of LLZ-NP and LLZ-PEP-NP was more than 85%, while LUT-PEP-NP showed higher lutein entrapment efficiency because of the better capacity of peptides bound with lutein. Nanoencapsulation of lutein into LLZ-PEP-NP resulted in a significantly higher solubility compared to nanoencapsulation of lutein into LLZ-NP and free lutein. The stabilities of lutein in zein-derived peptide nanoparticles in simulated gastric fluid (SGF) and simulated intestinal fluid (SIF) were improved. These results suggest that zein-derived peptides have the potential to be used as nanocarriers to enhance the solubility and stability of lutein, which can further improve its bioavailability.

The focus of the present work was to investigate the glycosylation of zein, partial properties of the glycosylated zein (GZ) and its retarding effect on lipid oxidation of ground pork. Zein was glycosylated with chitosan (MW 1500Da) by microbial transglutaminase, the reaction was verified by FT-IR. Under the optimized conditions, 97.48mg of glucosamine was covalently conjugated to 1g of zein, determined by HPLC. The solubility and the surface hydrophobicity of GZ were significantly improved. In vitro studies of GZ showed a dose-dependent scavenging activity against free radicals of DPPH, superoxide and hydroxyl radical, and the EC50 value for DPPH radical was 1.99μg TE/mg protein. In addition, reducing power and Fe2+-chelating capacity of it were 16.60 and 12.96μg TE/mg protein, respectively. GZ resulted in low levels of thiobarbituric acid-reactive substances and peroxide value of ground pork. These results suggest that GZ is a potential natural antioxidant.

Neoalbaconol, derived from Albatrellus confluens, shows anti-cancer activities in the previously study, but its role in angiogenesis is unknown. Here, we determined whether neoalbaconol could attenuate angiogenesis and how does it occur. Data demonstrated that neoalbaconol could inhibit the proliferation of breast cancer cells and induce apoptosis. Also, neoalbaconol suppressed vascular endothelial growth factor (VEGF)-induced human umbilical vascular endothelial cells (HUVECs) proliferation, migration, invasion, and capillary-like tube formation in vitro and reduced tumor angiogenesis in vivo. VEGF receptor activation and the downstream signal transduction cascades activation were inhibited by neoalbaconol. Additionally, neoalbaconol blocked EGFR-mediated VEGF production. EGFR overexpression reversed the neoalbaconol-induced VEGF reduction, confirming the importance of the EGFR inhibition in anti-angiogenesis of neoalbaconol. Furthermore, neoalbaconol inhibited tumor growth and tumor angiogenesis in a breast cancer xenograft model in vivo. Taken together, these results indicate that neoalbaconol could inhibit tumor angiogenesis and growth through direct suppression effects on vascular endothelial cells and reduction of proangiogenic factors in cancer cells.

New glycosylated zein peptides (GZP) were produced by transglutaminase-induced d -glucosamine conjugation onto zein peptides. GZP’s antagonistic effects on alcohol-induced liver injury in rats were evaluated. Compared with the alcohol model group, GZP (250 mg/kg·bw) remarkably increased alcohol dehydrogenase, acetaldehyde dehydrogenase, endogenous antioxidant enzymes activities, and GSH levels in liver, decreased serum triacylglycerol, tumor necrosis factor-α, liver malonaldehyde, reactive oxygen species, and lipopolysaccharide (LPS) levels, and significantly reversed pathological changes in liver tissues. These results indicate that low-dose GZP administration can alleviate alcohol-induced liver injury by accelerating alcohol metabolism, reversing hepatic redox status, and attenuating LPS-mediated inflammation responses. GZP is a potential alcohol metabolism promoter and liver-protective agent.

Wounds infected by bacteria are dangerous for human beings. However, along with the emergence of new strains and strong bacterial resistance, traditional antibiotics are unable to meet the medical needs for treating bacterial infections. Thus, new antibacterial substances with superior antimicrobial properties are urgently needed. Herein, a hydrogel containing poly acrylic acid (PAA), glycerol and Cu-MOP (named PAA-Cu-MOP hydrogel) is obtained by a facile mixing and ultrasonic procedure for wound healing treatment. This PAA-Cu-MOP hydrogel with high biocompatibility exhibits excellent wound healing behavior and is even better than the one of recombinant human epidermal growth factor. Tissue experiment results reveal that the PAA-Cu-MOP hydrogel accelerates the wound healing process by promoting angiogenesis, stimulating cell proliferation, and up-regulating cell factors.

This study compared two immobilization matrices like calcium-alginate and chitosan for immobilization of α-galactosidase and evaluated their potential for the removal of non-digestible raffinose family oligosaccharides from soy milk which cause abdominal discomfort. The pH optima of the free and immobilized enzymes were found to be similar at pH 4.0. The chitosan-immobilized α-galactosidase displayed higher optimal temperature (60°C) compared to alginate-immobilized enzyme (45°C) and free enzyme (50°C). The chitosan-immobilized and alginate-immobilized α-galactosidases displayed 93.7% and 97.6% hydrolysis of raffinose family oligosaccharides, respectively, while the free enzyme hydrolyzed only 30.3% oligosaccharides present in soy milk in 4 hr. Remarkably, both the immobilized enzymes showed complete removal of raffinose family oligosaccharides in 8 hr. Moreover, reusability studies indicate that even after five cycles of reuse, the chitosan and alginate-immobilized enzymes displayed 99% and 60% hydrolysis, respectively. PRACTICAL APPLICATIONS: In this study, we have used two inexpensive and non-toxic matrices for immobilizing α-galactosidase. We report that entrapment of α-galactosidase with chitosan significantly improved the optimal temperature of α-galactosidase, which is advantageous in food industry. The hydrolysis of raffinose family oligosaccharides in soy milk was also greatly enhanced after immobilization with chitosan and alginate. Thus, the results described in this study have relevance for development of safe, cost-effective and efficient method for removal of non-digestible soy oligosaccharides in food industry.

Lead halide perovskites have attracted much attention following their successful use in photovoltaic devices with impressive power-conversion efficiencies. Owing to perovskite quantum dots’ ionic nature, the surface ligands are easy to be overdamaged in polar solvents. Thus, there have been some difficulties in protecting the surface ligands of the quantum dots during purification processes, especially in solvent reprecipitation techniques. Here, we used mixtures of ethyl acetate and methyl acetate during the purification process to precipitate perovskite quantum dots that showed scarcely any damage to their surface ligands; furthermore, these purified CsPbBr3 quantum dots displayed high performances in devices. Quantum dot films were prepared via spin-coating from solution for use in solar cells with a simple structure (indium tin oxide/ZnO/quantum dots/Au), which displayed power-conversion efficiencies of up to 2.43%. Multiple purification cycles of this purification process allowed us to obtain near-perfectly modified perovskite colloidal quantum dots, ideal for application in optoelectronic devices.

Objective This study was to establish a rapid quantitative method for determination of six characteristic flavonoids, including licochalcone A (1), liquiritigenin (2), glabridin (3), isoliquiritigenin (4), liquiritoside (5), and isoliquiritoside (6) in licorice by a ultra-performance convergence chromatography (UPC2) apparatus equipped with a photodiode-array detector (PDA). Methods: The flavonoids studied were separated on an ACQUITY UPC2 TM Torus 2-PIC column (2.1 × 100 mm, 1.7 μm) using gradient elution (85:15–70:30) with a mobile phase consisting of CO2 and methanol: formic acid (99.8:0.2, v/v), back pressure of 2,000 psi, flow rate of 0.8 mL/min, and detection at 254 nm. This method was further applied for the analysis of the flavonoids present in 10 batches of licorice roots samples. Results: Good linearity (R2 = 0.9990–0.9998) was achieved and limits of detection (LOD) and quantification (LOQ) ranging from 0.12 to 0.49 µg/mL and 0.46 to 1.61 µg/mL, respectively. The relative standard deviations (RSD) for repeatability and reproducibility were 2.30–2.80% and 2.30–2.98%, respectively. The flavonoids distributed differently in Glycyrrhiza uralensis , Glycyrrhiza glabra, and Glycyrrhiza inflate . Conclusion: The established rapid quantitative analysis method was environment friendly and suitable to the quality control of species licorice.

Monoliths composed of poly(1-vinylimidazole-ethylene glycol dimethacrylate), referred to as poly(VI-EGDMA), were modified with imidazole groups and used in capillary microextraction (CME) of trace levels of Co(II), Ni(II) and Cd(II) ions from human urine samples. The effects of sample pH, sample flow rate, sample volume, elution conditions and potentially interfering ions were investigated. Under the optimized conditions, the limits of detection (3σ) are 0.6 ng L−1 for Co(II), 6.4 ng L−1 for Ni(II) and 0.7 ng L−1 for Cd(II), with the relative standard deviations of 1.8 (Co), 4.2 (Ni) and 2.2% (Cd). A Certified Reference Material (ZK018–2, lyophilized human urine) was analyzed to validate the method, and the values found were in good agreement with the certified ones. The method was successfully applied to the analysis of trace Co, Ni and Cd in human urine samples, and the recoveries for spiked samples were in the range from 89% to 105%. The one-step preparation of monolithic material in our perception is an attractive feature for this material which also excels by good extraction performance in that it possesses abundant adsorption sites, good stability and a long lifetime.

A fibrinolytic enzyme was produced by the medicinal mushroom, Cordyceps militaris using submerged fermentation. The enzyme was purified from culture supernatant by hydrophobic interaction, ion exchange and gel filtration chromatographies. It was purified by 36 fold, with a specific activity of 1,467.4 U/mg protein and the final yield was 5.8%. The molecular weight of the enzyme as determined by SDS-PAGE and gel filtration was 28 kDa and 24.5 kDa, respectively, and its isoelectric point (pI) was 9.0 ± 0.2. It was found to be a glycoprotein with carbohydrate content of 1.67% (w/v). The enzyme was optimally active at 37 °C and pH 7.2. The enzyme activity was strongly inhibited by soybean trypsin inhibitor (SBTI) and aprotinin which indicated it to be a serine protease, while other inhibitors like N -α-tosyl- l -phenylalanine chloromethyl ketone (TPCK), phenyl methane sulfonyl fluoride (PMSF), pepstatin and metal chelator EDTA did not inhibit its activity. Amino acid sequences of the purified enzyme were determined partially by Q-TOF2 and they were IEDFPYQVDLR; ANCGGTVISEK; YVLTAGHCAEGYTGLNIR; TNYASVTPITADMICAGFPEGK; KDSCSGDSGGPLVTGGK; VVGIVSFGTGCAR; ANKPGVYSSVASAEIR. Sequences of the seven peptides completely matched with those of a trypsin-like serine protease from Cordyceps militaris CM01 (accession no. EGX95217.1 ). The purified enzyme degraded α chains of fibrinogen first and then β and γ chains and also activated plasminogen into plasmin. It can act as an anticoagulant and prevent clot formation by degrading fibrinogen. Based on these studies, the purified enzyme has great potential to be developed as a natural agent for prevention and treatment of thrombolytic diseases.

In this work, a novel method of Au nanoparticles (NPs) modified polymer monolithic capillary microextraction (CME) on-line coupling with inductively coupled plasma-mass spectrometry (ICP-MS) was developed for the determination of trace rare earth elements (REEs) in environmental and biological samples. The poly(glycidyl methacrylate-ethylene dimethacrylate) monolithic capillary was prepared, functionalized with thiol ligands for the attaching of Au NPs, and then modified with mercaptosuccinic acid to provide massive carboxyl groups which have high affinity to REEs. With the modification of Au NPs, the adsorption capacity of the monolith towards target REEs has been improved by 3 to 6.5 times. Under the optimized conditions, the limits of detection of the developed method for REEs were in the range of 0.16 (Tb)–0.85 (Gd) ng L− 1. The enrichment factor was 25-fold with the sample throughput of 10 h− 1. And the relative standard deviations were between 2.7 (Lu) and 9.8% (Dy) (c = 10 ng L− 1, n = 9). The accuracy of the method was validated by the analysis of a standard stock solution of GSB04-1789-2004 and a Certified Reference Material of GBW07301a stream sediment. The proposed method was applied for the analysis of trace REEs in seawater samples as well as human whole blood with good recoveries. The prepared monolith is featured with strong anti-interference ability, superior adsorption capacity as well as long lifespan, and the developed monolithic CME-ICP-MS is sensitive, simple and rapid for the analysis of trace/ultra-trace REEs in environmental and biological samples with complex matrix.

The biochemical reactions involving trace elements depend on the nature and form of their chemical species in a biological system. Consequently, the study of the chemical speciation of a given element in biological samples has become essential for understanding their bioavailability and toxicity. Inductively coupled plasma-mass spectrometry (ICP-MS) hyphenated techniques are the most powerful and widely used for elemental speciation combining both high sensitivity and good selectivity for the determination of ultra-trace elements in biological samples. However, direct analysis of biological samples can suffer from matrix interferences, and both the low concentrations of chemical species, and in some cases, the limited amount of sample available can present analytical challenges. Therefore, there is a need to develop sample pre-treatment methods that can be used to remove the matrix and pre-concentrate the species of interest prior to analysis. This review will focus specifically on the state-of-the-art sample pre-treatment methods published in the last ten years including liquid and solid phase micro-extraction, and chip-based manifolds that can be coupled to ICP-MS hyphenated techniques. The different methods are evaluated for their ability to be deployed for the pre-treatment of actual biological samples. The development trends in this area of research are discussed, and integrated automated miniaturized systems are also reviewed.

The development of long-term imaging agents and subcellular imaging materials is of great importance in the research of cancer cell behaviors. In this work, a cationic poly( p -phenylenevinylene) derivative (PPV) is designed and synthesized to link quaternized N -methyl-imidazole groups as pendants which endow the polymer to bear positive charges. Absorption and fluorescence emission spectra of PPV display a large Stokes shift of 102 nm which is much larger than the commercial cell dyes. Positively charged polymer could adsorb onto the surface of cells via electrostatic interactions followed by cell endocytosis process to enter cells. Importantly, PPV barely has influence on the cell viability through cytotoxicity analysis. The colocalization data demonstrates that PPV and commercial lysosome-specific dye are highly colocalized in the same region, indicating that the green fluorescent PPV mainly distributes in the lysosomes. Moreover, the continuous imaging investigation shows that PPV could stay in cells for more than seven days while the commercial Lyso-Tracker would be extruded by cells after three days. PPV exhibits superior capabilities including strong fluorescence, large Stokes shift, good biocompatibility and high photostablity, which has great potential in the applications of cellular process monitoring.

A hydrogel was directly assembled from a Cu-MOP by a facile procedure without adding any polymers for the first time, and it exhibited excellent antibacterial activity towards both Gram-negative and Gram-positive bacteria.

Stereoisomers play an irreplaceable prominent role in life sciences. In molecular engineering, the isomers have been continuously designed and synthesized, and their interrelationships among structure-property-function have also been explored. However, the difficulties in purification and structural confirmation of tetraphenylethene (TPE) derivatives' stereoisomers hinder their structural diversity and scope of functions. Meanwhile, there were few researches on the macroscopic optical differences about TPE derivatives isomers. Herein, isoniazid-substituted TPE derivatives stereoisomers have been synthesized, as confirmed by single crystal data, mass spectrometry and NMR spectroscopy. The different photophysical properties were caused by the introduction of functional group isoniazid on the periphery of TPE. These isomers possessed dramatic differences in fluorescent properties, and exhibited diverse changes in response to stimuli. They also have distinct aggregated self-assembly morphologies because of different intermolecular hydrogen bonding, and they can be used as efficient fluorescent probes for detecting copper. Based on the coordination-driven sensing system, the morphologies of isomers aggregates changed significantly after interaction with copper, and the stoichiometry between them is 2:1. The theoretical calculation demonstrated that they have obvious intramolecular charge transfer behavior. Our research provided a good strategy for designing different substituted groups that can produce various functions and have more potential applications.

In this study, a fungal and two yeast β-galactosidases were immobilized using alginate and chitosan. The biochemical parameters and lactose hydrolysis abilities of immobilized enzymes were analyzed. The pH optima of immobilized fungal β-galactosidases shifted to more acidic pH compared to free enzyme. Remarkably, the optimal temperature of chitosan-entrapped yeast enzyme, Maxilact, increased to 60 °C, which is significantly higher than that of the free Maxilact (40 °C) and other immobilized forms. Chitosan-immobilized A. oryzae β-galactosidase showed improved lactose hydrolysis (95.7%) from milk, compared to the free enzyme (82.7%) in 12 h. Chitosan-immobilized Maxilact was the most efficient in lactose removal from milk (100% lactose hydrolysis in 2 h). The immobilized lactases displayed excellent reusability, and chitosan-immobilized Maxilact hydrolyzed > 95% lactose in milk after five reuses. Compared to free enzymes, the immobilized enzymes are more suitable for cost-effective industrial production of low-lactose milk due to improved thermal activity, lactose hydrolysis efficiencies, and reusability.