Development of a method to screen and isolate xanthine oxidase inhibitors from black bean in a single step: Hyphenation of semipreparative liquid chromatography and stepwise flow rate countercurrent chromatography.

Black bean, in which isoflavones are the main active constituent, also contains saponins and monoterpenes. Soybean isoflavone is a secondary metabolite that is formed during the growth of soybean; it exhibits antioxidant and cardiovascular activities and traces estrogen-like effects. In this study, black bean isoflavones were extracted with n-butanol, and ultrafiltration-liquid chromatography-mass spectrometry was used to screen their activity. Subsequently, the inhibitors were isolated and purified using semipreparative liquid chromatography and stepwise flow rate countercurrent chromatography. Thereafter, five active compounds were identified using mass spectrometry and nuclear magnetic resonance experiments. Finally, the inhibition types of the xanthine oxidase inhibitors were determined using enzymatic kinetic studies. The IC50 values of daidzin, glycitein-7-O-glucoside, genistin, daidzein, and genistein were determined to be 35.08, 56.22, 30.76, 68.79, and 95.37 µg/mL, respectively. Daidzin, genistin, and daidzein exhibited reversible inhibition, whereas glycitein-7-O-glucoside and genistein presented irreversible inhibition. This novel approach, which was based on ultrafiltration-liquid chromatography-mass spectrometry and stepwise flow rate countercurrent chromatography, is a powerful method for screening and isolating xanthine oxidase inhibitors from complex matrices. The study of enzyme inhibition types is helpful for understanding the underlying inhibition mechanism. Therefore, a beneficial platform was developed for the large-scale production of bioactive and nutraceutical ingredients.

Prognostic impact of tertiary lymphoid structures in breast cancer prognosis: a systematic review and meta-analysis.

BACKGROUND: Tertiary lymphoid structures (TLSs), organizationally resemble lymph nodes, are frequently present in breast cancer (BCa). It is usually, but not always, associated with a positive prognosis or immunotherapy response in cancer patients. This meta-analysis was performed to assess the prognostic and clinical impact of TLSs in BCa. METHODS: We conducted a systematic search in PubMed, Embase, Cochrane Library, Web of Science, China National Knowledge Infrastructure, and WanFang Database to obtain eligible research data up to May 30, 2021. This meta-analysis is focusing on the studies evaluated the prognostic value of TLSs and the associated clinicopathologic indicators, related gene expression and survival. STATA software 16.0 software was used to assess the prognostic significance and clinical impact of TLSs. RESULTS: Nine studies involved with 2281 cases were incorporated in this meta-analysis, in which four of them evaluated the prognostic value of TLSs. There are 6 studies assessed the relationship of TLSs and 4 studies investigated the clinicopathologic parameters as well as the key gene expression, respectively. The results showed the presence of TLSs were predicting a better OS (HR = 0.61, 95% CI: 0.51-0.73, p < 0.001) and DFS (HR = 0.40, 95% CI: 0.17-0.93, p < 0.001) of BCa patients. It also revealed that the presence of TLSs was significantly correlated with tumor differentiation (p < 0.001), pTNM stage (p < 0.001), lymph node metastasis (p < 0.001), and TILs density (p < 0.001) of BCa, and the expression of Her2 (p < 0.001), ER (p < 0.001), PR (p < 0.001) and Ki67 (p = 0.009) of the tumor cell. CONCLUSION: Our results indicated that high levels of TLSs could predict a favorable prognosis for BCa. Moreover, the TLSs were significantly correlated with the clinicopathological indicators and the critical gene expression of BCa, indicating its potential clinical impact on BCa patients.

Aqueous Protein-Polymer Bioconjugation via Photoinduced RAFT Polymerization Using High Loading Heterogeneous Catalyst.

Light-driven polymerization, such as photoinduced electron/energy transfer-reversible addition-fragmentation chain transfer (PET-RAFT) polymerization, enables biological benign conditions and versatile functional polymer structure design, which is readily used in protein-polymer bioconjugates. However, conventional metalloporphyrinic homogeneous catalysts for PET-RAFT polymerization suffer from limited aqueous solubility and tedious purification. Here we demonstrate the design of PET-RAFT photocatalyst from the reticular assembled Zr-porphyrinic metal-organic frameworks (MOFs), along with a biomacromolecule-based chain transfer agent, as efficient bioconjugation tools in water. Our methodology offers manufacturing advantages on bioconjugates under mild conditions such that MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) and cytotoxicity assays have shown the preservation of the protein integrity, bioactivity, and high cell viability after PET-RAFT polymerization. We find that the fast kinetics are benefiting from the ultrahigh loading of metalloporphyrins in MOF-525-Zn. This heterogeneous catalyst also allows us to maintain living characteristics to incorporate myriads of monomers into block copolymers. Other advantages like easy postreaction purification, reusability, and high oxygen tolerance even in an open system are demonstrated. This study provides a tool of highly efficient heterogeneous photocatalysts for polymer-protein bioconjugation in aqueous media and paves the road for biological applications.

Magnetic Janus nanocomposites with iridium(iii) complexes for heterogeneous catalysis of logic controlled RAFT polymerization using multiplexed external switching.

Photoinduced electron/energy transfer-reversible addition-fragmentation chain transfer (PET-RAFT) polymerization has emerged as a versatile and highly-efficient method for the polymerization of more activated monomers including N,N-dimethylacrylamide and methyl acrylate, and less activated monomers including N-vinylpyrrolidone and vinyl acetate, whilst imposing composition, sequence and spatiotemporal regulation. Although significant progress has been achieved in terms of ability to regulate PET-RAFT polymerization through the implementation of myriad environmental cues, it is still a great challenge to introduce multiple external switches within a single catalyst to accomplish logic toggling of controlled radical polymerization (CRP). Herein, we report the synthesis and characterization of Fe3O4@aSiO2@PNMIr Janus nanocomposites coupled with immobilized heteroleptic iridium(iii) complexes for heterogeneous catalysis of PET-RAFT polymerization. With this catalytic nanoarchitecture, we demonstrate multi-stimuli switching of CRPs using three different external physical manipulations: light "ON"/"OFF", magnet "OUT"/"IN" and temperature "LOW"/"HIGH". In addition, these magnetic Janus nanocomposites endowed radical polymerization with various attractive characteristics such as compatibility of myriad monomer formulations including "more activated" and "less activated" monomers, unique oxygen tolerance and ppm-level catalyst dosage. Logic-controlled polymerization with Fe3O4@aSiO2@PNMIr nanocomposites provides a straightforward, robust and user-friendly strategy for realizing multiplexed external switching of polymer propagation using a single nanocatalyst without the involvement of exogenous reagents.

IL-27 is elevated in sepsis with acute hepatic injury and promotes hepatic damage and inflammation in the CLP model.

BACKGROUND: Immuno-inflammation plays an important role in the pathophysiological process of sepsis-associated acute hepatic injury (AHI). Interleukin 27 (IL-27) is an important inflammatory regulator; however, its role in this condition is not clear. METHODS: The clinical data and IL-27 serum levels in sepsis patients with or without AHI were analysed. Classical caecal ligation puncture (CLP) models were established in wild-type (WT) and IL-27 receptor (WSX-1)-deficient (IL-27R-/-) mice. In addition, exogenous IL-27 was injected into these mice, and the levels of IL-27, IL-6, and tumour necrosis factor alpha (TNF-α) in the serum and liver were then measured by enzyme-linked immunoassay (ELISA), quantitative PCR, and Western blotting. The severity of liver damage was evaluated by haematoxylin and eosin staining of liver tissue, TUNEL assay and evaluation of alanine aminotransferase (ALT) and aspartate transaminase (AST) serum levels. Furthermore, the effects of IL-27 on the levels of phosphorylated c-Jun N-terminal kinase (JNK) in macrophages were assessed by Western blotting, and the effects of IL-27 on the expression of IL-6 and TNF-α in macrophages were assessed by ELISA. RESULTS: IL-27 was elevated in sepsis patients with acute hepatic injury, which correlated with the Acute Physiologic Assessment and Chronic Health Evaluation II (APACHEII) scores, Sequential Organ Failure Assessment (SOFA) scores, and procalcitonin, C-reactive protein, IL-6, and TNF-α expression. In the CLP-WT group, IL-27 was highly expressed in the serum and liver, which correlated with the elevated content of ALT, AST, TNF-α, IL-6, and p-JNK in the serum and liver and the pathological injury of the liver. In CLP-IL-27R-/- group, however, the levels of ALT, AST, TNF-α, IL-6, and p-JNK in the serum and liver and the pathological injury of the liver were decreased. Treatment with exogenous IL-27 led to a further increase in these cytokines in WT mice after CLP. IL-27 treatment and lipopolysaccharide stimulation in vitro increased the expression of p-JNK, IL-6, and TNF-α in macrophages, and these changes were decreased by a JNK signalling pathway inhibitor. CONCLUSION: IL-27 is elevated in sepsis patients, especially those with acute hepatic injury. In addition, IL-27 can promote inflammatory reactions in the CLP-induced hepatic injury mice model.

Career-Specific Parenting Practices and Career Decision-Making Self-Efficacy Among Chinese Adolescents: The Interactive Effects of Parenting Practices and the Mediating Role of Autonomy.

This study examined the unique and interactive effects of various career-specific parenting practices (i.e., parental career support, interference, and lack of engagement) on Chinese high school students' career decision-making self-efficacy (CDSE) as well as the mediating role of autonomy in such associations. Based on data from 641 Chinese high school students (47.6% male; mean age = 15.28 years old, SD = 0.49) in 2016, two moderated mediating effects were identified. Higher level of parental career engagement strengthened the positive association between parental career support and adolescents' autonomy, which in turn, was associated positively with adolescents' CDSE. Parental career interference related negatively with adolescents' CDSE via autonomy when lack of parental career engagement was low, but related positively with adolescents' CDSE via autonomy when lack of parental career engagement was high. These findings advance our understanding of the underlying processes between career-specific parenting practices and adolescents' CDSE. Implications for practices were discussed.

A novel flexible nanogenerator made of ZnO nanoparticles and multiwall carbon nanotube.

In this paper, a novel flexible nanogenerator (FNG) made of zinc-oxide (ZnO) nanoparticles (NPs) and multiwall-carbon nanotubes (MW-CNTs) is presented. In this structure, ZnO NPs and MW-CNTs are mixed with polydimethylsiloxane (PDMS) uniformly to form an entire flexible nanogenerator. Serial tests illustrate that the output voltage and power density are as high as 7.5 V and 18.75 µW per cycle, respectively. Furthermore, by foot stamp on the FNG, a peak voltage as high as 30 V can be generated. Comparing to the control samples, it is also proved that adding MW-CNTs into the matrix could significantly enhance the output voltage from 0.8 to 7.5 V. In summary, our work indicates that the realization of flexible nanogenerators made of ZnO NPs and MW-CNTs is technologically feasible, which may bring out some important and interesting applications in energy harvesting.