Synthesis of triazole AD-1 derivatives and its mechanism of mediating DNA damage of ROS in lung cancer cells.

Based on the significant biological activities and the remarkable physical and chemical properties of 1H-1,2,3-triazole pharmacophore, we herein adopted the strategy of click chemistry to combine the triazole fragment and the unique scaffold of 25-OCH3-PPD (AD-1) to design a series of potent compounds inducing apoptosis and DNA damage. The anti-proliferative effect was verified by MTT assay and colony formation assay. DNA double-stand breaks (DSBs) were obtained by observing the nuclear focus formation and the protein expression of γ-H2AX. Cell cycle arrest was evaluated by the cycle-related proteins such as CDK2, CDK4, CDK6, Cyclin D1 and P21. Apoptosis was assessed by flow cytometry, mitochondrial membrane potential (MMP) detection and the expression of apoptosis-related proteins. Reactive oxygen species (ROS) generation was measured with 2', 7'-dichlorofluorescein diacetate (DCFH-DA) staining. According to SAR analysis, the most potent compound 6a exhibited great inhibitory effect against A549 cells, which IC50 value of 2.84 ± 0.68 µM. Furthermore, 6a remarkably induced DNA damage, cell cycle arrest and apoptosis in A549 cells. 6a treatment increased the levels of ROS. Network pharmacology and molecular docking predicted the potential signaling pathways and ligand-receptor interactions, and the results of western blotting showed that 6a inhibited the PI3K/Akt/Bcl-2 signaling pathway by decreasing PI3K and Bcl-2 and total level of Akt expression, while Bax and Cyt c were increasing in 6a-treated A549 cells. As mentioned above, 6a has a potent inhibitory effect in A549 cells through induction of DNA damage, apoptosis via ROS generation and modulation of PI3K/Akt/Bcl-2 signaling pathway.

Gypensapogenin H suppresses tumor growth and cell migration in triple-negative breast cancer by regulating PI3K/AKT/NF-κB/MMP-9 signaling pathway.

Due to its aggressiveness and high metastasis rates, triple-negative breast cancer (TNBC) is a ubiquitous and deadly disease for the majority of women globally. Gypensapogenin H (GH) is a novel dammarane-type triterpene isolated from hydrolyzate of total saponins from Gynostemma pentaphyllum. Our previous work demonstrated that GH promoted apoptosis in TNBC. In the present study, xenograft TNBC models (xenotransplantation of MDA-MB-231 cells in nude mice) were used to evaluate the efficacy of GH in vivo. We preliminarily predicted the mechanism of GH inhibiting breast cancer tumors at the gene level through transcriptome screening. Through western blot analysis of tumor tissue, we found that GH could inhibit tumor proliferation and migration by regulating the PI3K/AKT/NF-κB/MMP-9 signaling pathway in vivo. We also analyzedthe results at the cell level in vitro, which were consistent with those in vivo. In summary, GH inhibited TNBC growth in vivo and suppressed TNBC cell migration in vitro. Our findings could help understand the mechanism of action of GH and suggest that GH would be a promising agent for TNBC therapy.

Gypensapogenin I Ameliorates Isoproterenol (ISO)-Induced Myocardial Damage through Regulating the TLR4/NF-κB/NLRP3 Pathway.

Myocardial fibrosis (MF) is a common pathological feature of many heart diseases and seriously threatens the normal activity of the heart. Jiaogulan (Gynostemma pentaphyllum) tea is a functional food that is commercially available worldwide. Gypensapogenin I (Gyp I), which is a novel dammarane-type saponin, was obtained from the hydrolysates of total gypenosides. It has been reported to exert a beneficial anti-inflammatory effect. In our study, we attempted to investigate the efficiency and possible molecular mechanism of Gyp I in cardiac injury treatment induced by ISO. In vitro, Gyp I was found to increase the survival rate of H9c2 cells and inhibit apoptosis. Combined with molecular docking and Western blot analysis, Gyp I was confirmed to regulate the TLR4/NF-κB/NLRP3 signaling pathway. In vivo, C57BL6 mice were subcutaneously injected with 10 mg/kg ISO to induce heart failure. Mice were given a gavage of Gyp I (10, 20, or 40 mg/kg/d for three weeks). Pathological alterations, fibrosis-, inflammation-, and apoptosis-related molecules were examined. By means of cardiac function detection, biochemical index analysis, QRT-PCR monitoring, histopathological staining, immunohistochemistry, and Western blot analysis, it was elucidated that Gyp I could improve cardiac dysfunction, alleviate collagen deposition, and reduce myocardial fibrosis (MF). In summary, we reported for the first time that Gyp I showed good myocardial protective activity in vitro and in vivo, and its mechanism was related to the TLR4/NF-κB/NLRP3 signaling pathway.

Improving rice eating and cooking quality by coordinated expression of the major starch synthesis-related genes, SSII and Wx, in endosperm.

KEY MESSAGE: Coordinated regulation of amylose and amylopectin synthesis via manipulation of SSII-2, SSII-3 and Wx expression in endosperm can improve rice eating and cooking quality. With increasing rice consumption worldwide, many researchers are working to increase the yield and improve grain quality, especially eating and cooking quality (ECQ). The rice ECQ is mainly controlled by the expression of starch synthesis-related genes (SSRGs) in endosperm. Although the Wx and SSII-3/SSIIa/ALK genes, two major SSRGs, have been manipulated to improve rice ECQ via various breeding approaches, new methods to further improve ECQ are desired. In our previous study, we enhanced rice ECQ by knocking down SSII-2 expression in the japonica Nipponbare cultivar (carrying the Wxb allele) via RNA interference. Herein, the SSII-2 RNAi was introduced into two Nipponbare-derived near-isogenic lines (NILs), Nip(Wxa) and Nip(wx), carrying Wxa and wx alleles respond for high and no amylose levels, respectively. Analysis of physicochemical properties revealed that the improved grain quality of SSII-2 RNAi transgenic lines was achieved by coordinated downregulating the expression of SSII-2, SSII-3 and Wx. To further confirm this conclusion, we generated ssii-2, ssii-3 and ssii-2ssii-3 mutants via CRISPR/Cas9 technique. The amylopectin structure of the resulting ssii-2sii-3 mutants was similar to that in SSII-2 RNAi transgenic lines, and the absence of SSII-2 decreased the amylose content, gelatinisation temperature and rapid visco-analyser profile, indicating essential roles for SSII-2 in the regulation of amylopectin biosynthesis and amylose content in rice endosperm. The effect of SSII-2 was seen only when the activity of SSII-3 was very low or lacking. Our study provides novel approaches and valuable germplasm resources for improving ECQ via plant breeding.

[Incidence of neonatal asphyxia and contributing factors for the develpment of severe asphyxia in Hubei Enshi Tujia and Miao Autonomous Prefecture: a multicenter study].

OBJECTIVE: To investigate the incidence of neonatal asphyxia and possible contributing factors for the development of severe asphyxia in Hubei Enshi Tujia and Miao Autonomous Prefecture, China. METHODS: A total of 16 hospitals in Hubei Enshi Tujia and Miao Autonomous Prefecture were selected as research centers. A retrospective analysis was performed for the clinical data of 22 294 live births in these 16 hospitals from January to December, 2016 to investigate the incidence rate of neonatal asphyxia and possible contributing factors for the development of severe asphyxia. RESULTS: Of the 22 294 neonates born alive, 733 (3.29%) were diagnosed with neonatal asphyxia, among whom 627 had mild asphyxia and 106 had severe asphyxia. The neonates with low maternal education level, maternal anemia during pregnancy, chorioamnionitis, abnormal amniotic fluid, abnormal umbilical cord, placenta previa, placental abruption, Tujia Minority, preterm birth, and low birth weight had a higher incidence of severe asphyxia (P<0.05). CONCLUSIONS: The incidence rate of neonatal asphyxia in Hubei Enshi Tujia and Miao Autonomous Prefecture is higher. Low maternal education level, maternal anemia during pregnancy, chorioamnionitis, abnormal amniotic fluid, abnormal umbilical cord, placenta previa, placental abruption, Tujia Minority, preterm birth, and low birth weight may be related to the development of severe neonatal asphyxia.

[Influence of environmental factors on the migratory birds population with chronic obstructive pulmonary disease].

OBJECTIVE: To explore the influence of environment factors on the migratory birds population with chronic obstructive pulmonary disease (MDCOPD). METHODS: A total of 82 stable MDCOPD patients from Beijing area, who went to Hainan Branch of PLA General Hospital and the People's Hospital of Sanya for treatment between September 2012 and May 2014, were recruited in this study.At Sanya, from October to April of the following year (wintertime), all the patients were evaluated by COPD assessment test (CAT) and modified medical research council (mMRC) questionnaire, and follow-up was performed every 2 months.From May to September (summertime), when the MDCOPD patients came back to Beijing area, the patients were evaluated by CAT and mMRC questionnaire again, and still follow-up was performed every 2 months.The scores of before and after living environment changed were compared.Acute exacerbations or hospitalizations were recorded if any occurred in the patients during the follow up period, and the patients were reevaluated by the two questionnaires 1 month later after they became stable.At the same time, the daily air quality index (AQI) from the national urban air quality publishing platform and the temperature and relative humidity from the local weather network were recorded. RESULTS: The score of CAT in Sanya was significantly lower than that in Beijing ((20.0 ± 8.0) vs (26.4 ± 7.1) scores, P < 0.001); the score of mMRC in Sanya was also significantly lower than Beijing ((1.9 ± 0.8) vs (2.9 ± 0.9) scores, P = 0.001). Based on the latest guidelines, the classification of COPD patients changed from D (in Beijing) to C (in Sanya) according to mMRC scores, while the CAT scores from the two cities did not cause the change of classification.The difference of temperature between wintertime of Sanya and summertime of Beijing was not significant ((23.1 ± 4.2) vs (24.3 ± 2.3) °C, P = 0.674); the wintertime's AQI in Sanya was significantly lower than the summertime's AQI in Beijing ((49.4 ± 8.9) vs (113.1 ± 14.2) µg/m³, P < 0.001); the relative humidity of wintertime in Sanya was significantly higher than that of summertime in Beijing ((90.6 ± 4.9)% vs ((58.0 ± 10.0)%, P = 0.001). CONCLUSION: When the MDCOPDs live in the area where has a better air quality and higher relative humidity, their quality of life and activity tolerance can be improved obviously.

Postbiotics from Pichia kudriavzevii promote intestinal health performance through regulation of Limosilactobacillus reuteri in weaned piglets.

Postbiotics are attractive as alternatives to antibiotics for use against post-weaning diarrhea. However, their beneficial mechanisms are largely unknown. In the current study, we first demonstrated that supplementation with 0.5% Pichia kudriavzevii FZ12 postbiotics in the diet significantly reduced diarrhea incidence, promoted growth performance, improved gut health performance, and significantly enriched beneficial bacteria, particularly Lactobacillus spp., in the intestines of weaned piglets. Importantly, we identified a heat- and proteinase K-sensitive component, cytochrome c, of the postbiotics that significantly promoted the growth and biofilm formation of Limosilactobacillus reuteri FP13. We demonstrated the importance of P. kudriavzevii FZ12 postbiotics in improving the intestinal health of a model animal and revealed that cytochrome c is one of the important components of yeast postbiotics. These findings may provide new insights into microbe-postbiotics interplay that can be applied to guidelines for dietary modulation to alleviate weaning-induced diarrhea.

A peroxyoxalate chemiluminescence recovery system based on the interaction of N-doped graphene oxide nanosheets and an oligopeptide for ultra-sensitive and selective copper(II) ion detection.

In this paper, a peroxyoxalate chemiluminescence (CL) recovery system based on the interaction of N-doped graphene oxide nanosheets (N-GONs) and an oligopeptide for copper(II) ion detection has been reported. N-GONs as an excellent CL enhancer are prepared by the hydrothermal method using citric acid and ammonia, and the morphology and structure are characterized in detail by TEM, XPS, FT-IR and UV/vis, etc. In the bis(2,4,6-trichlorophenyl)oxalate (TCPO) and hydrogen peroxide (TCPO + H2O2) CL reaction system, the addition of N-GONs gives a remarkable CL emission, which can be quenched by an oligopeptide composed of ten amino acid residues due to the interaction between the N-GON plane and the oligopeptide strand. While in the presence of copper(II) ion, the quenched CL is recovered gradually along with the addition of copper(II) ion in the system. Based on the above CL reactions, a TCPO + H2O2 + N-GONs + oligopeptide CL system is constructed, achieving an ultra-sensitive and selective detection of copper(II) ion in environmental water samples. The detection limit of this method is as low as 0.2 pmol L-1, which is at least three orders of magnitude lower than other CL methods. The N-GONs and oligopeptide involved in the CL system are environmentally friendly, making it possess potential in the detection of copper(II) ion in environmental water samples.

Gypensapogenin I Suppresses Cell Proliferation in Triple-Negative Breast Cancer Via Triggering the Closure of AKT/GSK3ß/ß-Catenin and Notch-1 Signaling Pathways.

Jiaogulan (Gynostemma pentaphyllum) tea is a functional food that is commercially available worldwide. Gypensapogenin I (Gyp I), which is a natural damarane-type saponin, was obtained from the hydrolysates of total gypenosides. The present research was performed to investigate the potential antiproliferation effect of Gyp I in MDA-MB-231 cells and the underlying mechanisms. Here, we found that Gyp I attenuated survival, inhibited proliferation, and induced apoptosis in MDA-MB-231 cells. Target prediction by binding molecule docking and western blot assays confirmed the mechanism by which Gyp I inhibited the proliferation of breast cancer cells via the AKT/GSK3ß/ß-catenin signaling pathway. We also showed that Gyp I exhibited superior in vivo efficacy that was dose dependent. Tumor tissue transcriptome analysis indicated that Gyp I could decrease the expression levels of NOTCH1 and HES1, which was in contrast to the effect on MAML and NUMBL, indicating that our compound hindered the activation of the Notch-1 signaling pathway. In summary, we report for the first time that Gyp I shows excellent anti-breast cancer activity in vivo and in vitro and that its pathway of action is related to the AKT/GSK3ß/ß-catenin and Notch-1 signaling pathways. Therefore, Jiaogulan tea can not only be used as a health food but also possesses the possibility to treat triple-negative breast cancer (TNBC).

Near-infrared light triggered multi-hit therapeutic nanosystem for tumor specific photothermal effect amplified signal pathway regulation and ferroptosis.

The high therapeutic resistance of tumor is the primary cause behind tumor recurrence and incurability. In recent years, scientists have devoted themselves to find a variety of treatments to solve this problem. Herein, we propose a multi-hit strategy that is based on the biodegradable hollow mesoporous Prussian blue (HMPB)-based nanosystem for tumor-specific therapy that encapsulated the critical heat shock protein 90 (HSP90) inhibitor 17-dimethylamino-ethylamino-17-demethoxydeldanamycin (17-DMAG). The nanosystem was further modified using thermotropic phase transition material star-PEG-PCL (sPP) and hyaluronic acid (HA), which offers near infrared light (NIR) responsive release characteristic, as well as enhanced tumor cell endocytosis. Upon cell internalization of 17-DMAG-HMPB@sPP@HA and under 808 nm laser irradiation, photothermal-conversion effect of HMPB directly kills cells using hyperthermia, which further causes phase transition of sPP to trigger release of 17-DMAG, inhibits HSP90 activity and blocks multiple signaling pathways, including cell cycle, Akt and HIF pathways. Additionally, the down-regulation of GPX4 protein expression by 17-DMAG and the release of ferric and ferrous ions from gradual degradation of HMPB in the endogenous mild acidic microenvironment in tumors promoted the occurrence of ferroptosis. Importantly, the antitumor effect of 17-DMAG and ferroptosis damage were amplified using photothermal effect of HMPB by accelerating release of ferric and ferrous ions, and reducing HSP90 expression in cells, which induced powerful antitumor effect in vitro and in vivo. This multi-hit therapeutic nanosystem helps provide a novel perspective for solving the predicament of cancer treatment, as well as a promising strategy for design of a novel cancer treatment nanoplatform.

A mesoporous silica-based probe with a molecularly imprinted polymer recognition and Mn:ZnS QDs@rhodamine B ratiometric fluorescence sensing strategy for the analysis of 4-nitrophenol.

In this study, a mesoporous silica fluorescence probe co-doped with manganese-doped zinc sulfide quantum dots (Mn:ZnS QDs) and rhodamine B (RB) and coated with molecularly imprinted polymer (MIP) has been prepared by sol-gel methods. The morphology and structure were characterized in detail by transmission electron microscopy (TEM), Fourier transform-infrared absorption spectroscopy (FT-IR) and ultraviolet-visible absorption spectroscopy (UV-vis). The probe exhibited two characteristic emission peaks at 411 nm and 582 nm, and the synchronous ratiometric fluorescence responses F411/F582 to different concentrations of 4-nitrophenol (4-NP) showed a good linear correlation in the range of 0.01-10 µmol L-1 besides achieving the sensitive detection of 4-NP with a detection limit as low as 3.0 nmol L-1 (3σ). The probe possesses the advantages of selectivity toward the target molecular structure, self-stability in the detection time domain and anti-interference ability, exhibiting excellent potential for application in 4-NP detection in different water environments.

Exploration of the Potential Transcriptional Regulatory Mechanisms of DNA Methyltransferases and MBD Genes in Petunia Anther Development and Multi-Stress Responses.

Cytosine-5 DNA methyltransferases (C5-MTases) and methyl-CpG-binding-domain (MBD) genes can be co-expressed. They directly control target gene expression by enhancing their DNA methylation levels in humans; however, the presence of this kind of cooperative relationship in plants has not been determined. A popular garden plant worldwide, petunia (Petunia hybrida) is also a model plant in molecular biology. In this study, 9 PhC5-MTase and 11 PhMBD proteins were identified in petunia, and they were categorized into four and six subgroups, respectively, on the basis of phylogenetic analyses. An expression correlation analysis was performed to explore the co-expression relationships between PhC5-MTases and PhMBDs using RNA-seq data, and 11 PhC5-MTase/PhMBD pairs preferentially expressed in anthers were identified as having the most significant correlations (Pearson's correlation coefficients > 0.9). Remarkably, the stability levels of the PhC5-MTase and PhMBD pairs significantly decreased in different tissues and organs compared with that in anthers, and most of the selected PhC5-MTases and PhMBDs responded to the abiotic and hormonal stresses. However, highly correlated expression relationships between most pairs were not observed under different stress conditions, indicating that anther developmental processes are preferentially influenced by the co-expression of PhC5-MTases and PhMBDs. Interestingly, the nuclear localization genes PhDRM2 and PhMBD2 still had higher correlations under GA treatment conditions, implying that they play important roles in the GA-mediated development of petunia. Collectively, our study suggests a regulatory role for DNA methylation by C5-MTase and MBD genes in petunia anther maturation processes and multi-stress responses, and it provides a framework for the functional characterization of C5-MTases and MBDs in the future.

Starch biosynthesis in cereal endosperms: An updated review over the last decade.

Starch is a vital energy source for living organisms and is a key raw material and additive in the food and non-food industries. Starch has received continuous attention in multiple research fields. The endosperm of cereals (e.g., rice, corn, wheat, and barley) is the most important site for the synthesis of storage starch. Around 2010, several excellent reviews summarized key progress in various fields of starch research, serving as important references for subsequent research. In the past 10 years, many achievements have been made in the study of starch synthesis and regulation in cereals. The present review provides an update on research progress in starch synthesis of cereal endosperms over the past decade, focusing on new enzymes and non-enzymatic proteins involved in starch synthesis, regulatory networks of starch synthesis, and the use of elite alleles of starch synthesis-related genes in cereal breeding programs. We also provide perspectives on future research directions that will further our understanding of cereal starch biosynthesis and regulation to support the rational design of ideal quality grain.

Study on the electrochemical behavior and differential pulse voltammetric determination of rhein using a nanoparticle composite film-modified electrode.

A sensitive electrochemical method was developed for the differential pulse voltammetric determination of rhein at a glassy carbon electrode (GCE) modified with a nanoparticle composite film. In the present paper, multi-wall carbon nanotube (MWNT) was dispersed into dihexadecyl phosphate (DHP) to give a homogeneous suspension. After the solvent evaporation, a uniform film of MWNT-DHP composite film was obtained on the GCE surface. The MWNT-DHP composite film-modified GCE exhibited excellent electrocatalytic behavior toward the redox of rhein. Compared with an irreversible reduction of rhein at the bare GCE, a reversible redox behavior of rhein was observed at the MWNT-DHP composite film-modified GCE and the redox current was also enhanced greatly. Based on this, a cathodic differential pulse voltammetry (DPV) was applied for the determination of rhein. The experimental parameters, which influence the current of rhein, were optimized. Under optimal conditions, the cathodic DPV measurements were performed and a linear response of rhein was obtained in the range from 1.0 x 10(-8) to 5.0 x 10(-6) mol L(-1) and with a limit of detect (LOD) of 5.0 x 10(-9) mol L(-1). The proposed procedure was successfully applied to assay rhein in real samples with satisfactory results.

[Screening of mimic epitopes of Toxoplasma gondii antigen by phage-displayed peptide library].

Specific antibodies purified from sera of rabbit infected with Toxoplasma gondii were used to immunoscreen the 12-mer and 7-mer phage random peptide libraries. By 3 rounds of screening, 70 clones were picked out randomly to test the specificity and 43 were found positive by ELISA. 23 of the 43 positive clones showing strong reaction in ELISA were picked out for sequencing. Clone A7 was selected as mimic antigen in ELISA test after analysis of the 23 short peptide sequences. 23 out of 47 sera from rabbits infected with Toxoplasma gondii were ELISA positive by A7 mimic antigen with a positive rate of 68.1%, and 10 of 155 sera from healthy persons showed false positive with a specificity of 93.5%. The primary result suggests that mimic antigen may have a potential use in diagnose of toxoplasmosis.