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Angiogenesis inhibition has become a promising therapeutical strategy for cancer treatment. Current clinical anti-angiogenesis treatment includes antibodies against vascular endothelial growth factor (VEGF) or VEGF receptor, fusion proteins with high affinity to VEGF receptor, and tyrosine kinase inhibitors of VEGF receptor. However, current treatments are prone to systemic toxicity or acquiring drug resistance. A natural bioactive lipid 1,2-dipalmitoyl-sn‑glycero-3-phosphate (dipalmitoyl phosphatidic acid, DPPA) was reported to exhibit anti-angiogenic and anti-tumoral activity. However, the hydrophobic property of DPPA largely restricted its clinical use, while systemic infusion of free DPPA could result in undesirable side effects. Herein, we successfully developed DPPA-based lipid-nanoparticles (DPPA-LNPs) which turns the "therapeutic payload into nanocarrier". This strategy could improve on DPPA's hydrophiliciy, thereby facilitating its systemic administration. . DPPA-LNPs not only retained the therapeutic anti-angiogenic and anti-tumoral bioactivity of parental DPPA, but also greatly improved its tumor targeting ability via enhanced permeability and retention (EPR) effect. This strategy not only eliminates the limitation of drug encapsulation rate, toxicity of the delivery vehicle; but also enhances DPPA bioacvtity in vitro and in vivo. Systemic administration of DPPA-LNPs significantly suppressed the blood vessel formation and tumor growth of triple negative breast cancer and liver cancer growth on both xenograft tumor models. STATEMENT OF SIGNIFICANCE: This is the first-in-kind self-therapeutic inherent lipid to be made into a nanocarrier, with inherent anti-angiogenic and anti-tumor properties. DPPA nanocarrier is fully natural, fully compatible with minimal systemic toxicity. DPPA nanocarrier can accumulate at high concentration at tumor via EPR effect, exerting both anti-angiogenic and anti-tumor effects in vivo. DPPA nanocarrier could be used to encapsulate biologics or small molecules for synergistic anti-cancer therapy.

Since the discovery of RNA interference (RNAi), RNAi technology has rapidly developed into an efficient tool for post-transcriptional gene silencing, which has been widely used for clinical or preclinical treatment of various diseases including cancer. Small interfering RNA (siRNA) is the effector molecule of RNAi technology. However, as polyanionic macromolecules, naked siRNAs have a short circulatory half-life (

During froth treatment, a water-in-diluted bitumen emulsion is obtained. The emulsified water contains chloride ions that form hydrochloric acid in downstream oil processing, leading to catalyst deactivation and equipment corrosion. Emulsified water, drops smaller than 10 μm, cannot be effectively removed by gravity settling and centrifugation to below 2 wt %. In this work, a filter-coalescer was used to promote the coalescence and separation of water-in-bitumen emulsion. The larger water drops (>300 μm) exiting the coalescer undergo gravity settling, reducing the water content in diluted bitumen emulsions to values lower than 0.1 vol %. The performance of the coalescer was interpreted via the colloid filtration theory of Rajagopalan and Tien (RT), improved in this work with a coalescence probability (CP) prefactor. This new RTCP framework was able to reproduce the experimental data, allowing its potential use as a predictive model for emulsion filtration and the operation of filter-coalescers. A capillary number analysis was used to account for the detachment of coalesced drops and interpret the drop sizes with different superficial velocities and bed porosities.

Structural morphology is the key parameter for efficacy of nanomedicine. To date, lipid-based nanomaterial has been the most widely used material in nanomedicine and many other biomedical applications. However, to the best of our knowledge, there has not been an in-depth or systematic investigation of the structure-function relationship of lipid-based nanostructures. In this report, we investigated the formulation of novel lipid-based nanostructures via simple tuning of lipid combinations. To prove this concept, we used a combination of various ratios of simple and common phospholipids with different chain lengths (14-carbon chain DMPC: 6-carbon chain DHPC) to find out whether a myriad of novel lipid nanostructures could be obtained. Interestingly, many combinations resulted in distinct lipid nanostructures. Drug encapsulation tests confirmed that they are able to load large amounts of drugs for biological application. In vivo anti-tumor efficacy revealed that certain lipid nanostructures possessed superior tumor retardation effects.

The expression of Proline-, glutamic acid-, and leucine-rich protein 1 (PELP1) has been reported to be dysregulated in non-small cell lung carcinoma, especially in lung adenocarcinoma (LUAD). Therefore, we aimed to investigate the functional and prognostic roles of PELP1 in LUAD in this study. We first immunolocalized PELP1 in 76 cases of LUAD and 17 non-pathological or tumorous lung (NTL) tissue specimens and correlated the findings with the clinicopathological parameters of the patients. We then performed in vitro analysis including MTT, flow cytometry, wound healing, and transwell assays in order to further explore the biological roles of PELP1 in 17-β-estradiol (E2) induced cell proliferation, migration, and invasion of LUAD cells. We subsequently evaluated the prognostic significance of PELP1 in LUAD patients using the online survival analysis tool Kaplan-Meier Plotter. The status of PELP1 immunoreactivity in LUAD was significantly higher than that in the NTL tissues and significantly positively correlated with less differentiated features of carcinoma cells, positive lymph node metastasis, higher clinical stage as well as the status of ERα, ERβ, and PCNA. In vitro study did reveal that E2 promoted cell proliferation and migration and elevated PELP1 protein level in PELP1-high A549 and H1975 cells but not in PELP1-low H-1299 cells. Knock down of PELP1 significantly attenuated E2 induced cell proliferation, colony formation, cell cycle progress as well as migration and invasion of A549 and H1975 cells. Kaplan-Meier Plotter revealed that LUAD cases harboring higher PELP1 expression had significantly shorter overall survival. In summary, PELP1 played a pivotal role in the estrogen-induced aggressive transformation of LUAD and could represent adverse clinical outcome of the LUAD patients.

Summary Targeted regulation of mitochondrial gene expression is challenging due to the lack of a mitochondria-specific delivery system. We have previously developed various stimuli-responsive nanoparticle (NP)-based delivery systems to transport nucleic acids for regulation of target gene expression. This protocol describes the design and preparation of an NP platform for mitochondria-specific gene delivery (mito-NP). We use mito-NP in primary liver fibroblasts that are transplanted into mice. Mito-NP can be used to deliver various nucleic acid therapeutics and to treat mitochondria-regulated diseases. For complete details on the use and execution of this protocol, please refer to Zhao et al. (2020)., Graphical Abstract, Highlights • A protocol for mitochondria-specific gene delivery system is described • The delivery system achieves targeted regulation of mitochondrial gene expression • This protocol is applicable to mitochondria-specific delivery of various nucleic acids, Targeted regulation of mitochondrial gene expression is challenging due to the lack of a mitochondria-specific delivery system. We have previously developed various stimuli-responsive nanoparticle (NP)-based delivery systems to transport nucleic acids for regulation of target gene expression. This protocol describes the design and preparation of an NP platform for mitochondria-specific gene delivery (mito-NP). We use mito-NP in primary liver fibroblasts that are transplanted into mice. Mito-NP can be used to deliver various nucleic acid therapeutics and to treat mitochondria-regulated diseases.

A series of substituted benzaldehyde thiosemicarbazide compounds (1–7) were synthesized as xanthine oxidase (XO) inhibitors, and the interactions between substituted benzaldehyde thiosemicarbazide compounds (1–7) and XO were studied by ultraviolet spectroscopy, fluorescence spectroscopy, and molecular docking. It was found that the hydrogen bond and hydrophobicity were the main interactions between substituted benzaldehyde thiosemicarbazide compounds and XO, and introducing OH at the para position of the benzene ring and a Ph- or Me-group at the amino terminal of compound 4 increased the modifier's inhibitory activity. The results suggest that the newly introduced benzene ring interacted with the hydrophobic cavity of XO by means of the π-π stacking force between the newly introduced benzene ring and the aromatic amino acid residues, such as the Phe residue, which greatly increased the modifier's inhibitory activity. We conclude that introducing the Ph-group at the amino terminal of compound 4 and the OH group at the para position of the benzene ring was a good route to obtain novel XO inhibitors. Fluorescence spectroscopy assisted by 8-anilino-1-naphthalenesulfonic acid fluorescence probing and molecular docking were helpful for achieving a preliminary and relatively clear understanding of the interactions between target compounds and XO, which deserve further study.

Pancreatic cancer (PAC) is one of the most lethal malignant neoplasms with poor prognosis and high mortality. Emerging evidence has revealed that abnormal tumor lipid metabolism and tumor-associated macrophages (TAMs) significantly contribute to PAC development and progression. Therefore, concurrently reprogramming tumor lipid metabolism and regulating TAMs function could be a promising strategy for effective PAC therapy. Herein, we identified an important enzyme catabolizing lipids (monoacylglycerol lipase, MGLL) and a key receptor regulating macrophage phenotype (endocannabinoid receptor-2, CB-2) that are over-expressed in PAC cells and on TAMs, respectively. Based on this finding, we developed a reduction-responsive poly (disulfide amide) (PDSA)-based nanoplatform for systemic co-delivery of MGLL siRNA (siMGLL) and CB-2 siRNA (siCB-2). This nanoplatform could utilize its reduction-responsive characteristic to rapidly release siRNA for efficient silencing of MGLL and CB-2, inducing concurrent suppression of free fatty acids (FFAs) generation in PAC cells and repolarization of TAMs into tumor-inhibiting M1-like phenotype. With this suppressed FFAs generation to inhibit nutrient supply for tumor cells and repolarized TAMs to secrete tumoricidal cytokines such as TNF-α and IL-12, a combinational anticancer effect could be achieved in both xenograft and orthotopic PAC tumor models.

A collection of thirty-six aromatic heterocycle thiosemicarbazone analogues presented a broad span of anti-tyrosinase activities were designed and obtained. A robust and reliable two-dimensional quantitative structure–activity relationship model, as evidenced by the high q 2 and r 2 values (0.848 and 0.893, respectively), was gained based on the analogues to predict the quantitative chemical-biological relationship and the new modifier direction. Inhibitory activities of the compounds were found to greatly depend on molecular shape and orbital energy. Substituents brought out large ovality and high highest-occupied molecular orbital energy values helped to improve the activity of these analogues. The molecular docking results provided visual evidence for QSAR analysis and inhibition mechanism. Based on these, two novel tyrosinase inhibitors O04 and O05 with predicted IC 50 of 0.5384 and 0.8752 nM were designed and suggested for further research.

Novel Tyrosinase Inhibitors of 4-functionalized Thiophene-2-carbaldehyde thiosemicarbazone (TCT) derivatives (1-8) had been synthesized and Spectrofluorimetry, 1H and 13C NMR titration and Molecular docking had been used to investigate their inhibitory activities and mechanisms on tyrosinase. The results showed that the inter-molecular interactions or hydrogen bond formation by increasing length of carbon chain or introducing benzene ring to the 4-functionalized ester group promoted or stabilized formation of complexes between modifier and tyrosinase, and enhanced the inhibitory activity of modifiers. The inhibitory activity of 4-benzoy methoxy-TCT was much stronger than that of any other synthesized tested modifiers, which was well explained by molecular docking and further verified by spectrofluorimetry and NMR titration by assuming that there existed an inter-molecular interaction besides formation of hydrogen bonds between the amino acid residues ASN260, GLU256, HIS85 of enzyme and the modifier.We concluded that 4-benzoy methoxy substitution of TCT was a good route obtaining novel tyrosinase inhibitors and deserved further studies.

We synthesized a series of 4- or 5-functionalized TCT derivatives (1-12) and investigated their inhibitory activities and mechanisms on tyrosinase by using Spectrofluorimetry, 1H and 13C NMR titration and IR spectra. The results of the fluorescence spectra and NMR titrations showed that the thiosemicarbazone moiety formed complexes with copper ions in the active center of the enzyme and played an important role in inhibiting the activities of the target compounds. The 5-functionalization decreased the inhibitory activity, but the 4-functionalization with a methoxyacetyl group enhanced the inhibitory activity, in which a strong auxiliary vicinity synergistic effect from the methoxyacetyl group strengthens and promotes the formation of complexes between the sulfur atoms of the inhibitor and the dicopper nucleus of tyrosinase. We concluded that the appropriate 4-functionalization improved the inhibitory activity of the modifier, and that 4-methoxyacetyl -TCT is promising for the inhibition of tyrosinase.

To analyse the damage of Q235A steel during axial tension, tensile experiments were carried out on several Q235A sheet specimens, the acoustic emission (AE) signals were collected at the same time. The properties of cumulative ringing counts of sheet specimens were analysed during tension process, the load-cumulative ringing counts curves were fitted, and the expression of steel damage factor was obtained by polynomial cumulative ringing counts. The cumulative ringing counts calculated by this expression under different loads are close to the experimental values, which prove that it is feasible to use the cumulative ringing counts of acoustic emission to represent the damage factors of Q235A steel during tensile process.

Background Compounds with the ability to scavenge reactive oxygen species (ROS) and inhibit tyrosinase may be useful for the treatment and prevention from ROS-related diseases. The number and location of phenolic hydroxyl of the flavonoids will significantly influence the inhibition of tyrosinase activity. Phenolic hydroxyl is indispensable to the antioxidant activity of flavonoids. Isoeugenol, shikonin, baicalein, rosmarinic acid, and dihydromyricetin have respectively one, two, three, four, or five phenolic hydroxyls. The different molecular structures with the similar structure to l-3,4-dihydroxyphenylalanine (l-DOPA) were expected to the different antityrosinase and antioxidant activities. Methods This investigation tested the antityrosinase activity, the inhibition constant, and inhibition type of isoeugenol, shikonin, baicalein, rosmarinic acid, and dihydromyricetin. Molecular docking was examined by the Discovery Studio 2.5 (CDOCKER Dock, Dassault Systemes BIOVIA, USA). This experiment also examined the antioxidant effects of the five compounds on supercoiled pBR322 plasmid DNA, lipid peroxidation in rat liver mitochondria in vitro, and DPPH, ABTS, hydroxyl, or superoxide free radical scavenging activity in vitro. Results The compounds exhibited good antityrosinase activities. Molecular docking results implied that the compounds could interact with the amino acid residues in the active site center of antityrosinase. These compounds also exhibited antioxidant effects on DPPH, ABTS, hydroxyl, or superoxide free radical scavenging activity in vitro, lipid peroxidation in rat liver mitochondria induced by Fe2+/vitamin C system in vitro, and supercoiled pBR322 plasmid DNA. The activity order is isoeugenol

Three phloretin derivates were achieved through condensation of the phloretin template with thiosemicarbazide, 4-methylthiosemicarbazide, and 4-phenyl-3-thiosemicarbazide. Their purity was confirmed by high-performance liquid chromatograph and their structures were determined from their ultraviolet spectra, Fourier-transform infrared, 1H and 13C NMR, and mass spectra. The solubility of these novel compounds in ultrapure water at 25 °C was significantly improved compared with that of phloretin in the following order: phloretin thiosemicarbazone (PT, >1.00 mg/mL) > phloretin-4-phenyl-3-thiosemicarbazone (PPT, 0.52 mg/mL) > phloretin-4-methylthiosemicarbazone (PMT, 0.24 mg/mL) > phloretin (0.02 mg/mL). The tyrosinase inhibitory effect and various antioxidant assays in vitro were evaluated. Results showed that PT possesses potent tyrosinase inhibitory activity (IC50 = 57.81 ± 1.46 μM), which was better than phloretin (IC50 = 70.08 ± 0.88 μM). They also significantly quenched 1,1-diphenyl-2-picryl-hydrazyl and exhibited strong activity against the lipid peroxidation induced by Fe2+/ascorbic acid. Likewise, these compounds significantly protected against 2,2′-azo-bis(2-amidinopropane)dihydrochloride-induced Cu, Zn-superoxide dismutase, and pBR322 plasmid DNA damage in a dose-dependent manner.

There is a clinical need for new, more effective treatments for chronic wounds in diabetic patients. Lack of epithelial cell migration is a hallmark of nonhealing wounds, and diabetes often involves endothelial dysfunction. Therefore, targeting re-epithelialization, which mainly involves keratinocytes, may improve therapeutic outcomes of current treatments. In this study, we present an integrin-binding prosurvival peptide derived from angiopoietin-1, QHREDGS (glutamine-histidine-arginine-glutamic acid-aspartic acid-glycine-serine), as a therapeutic candidate for diabetic wound treatments by demonstrating its efficacy in promoting the attachment, survival, and collective migration of human primary keratinocytes and the activation of protein kinase B Akt and MAPKp42/44. The QHREDGS peptide, both as a soluble supplement and when immobilized in a substrate, protected keratinocytes against hydrogen peroxide stress in a dose-dependent manner. Collective migration of both normal and diabetic human keratinocytes was promoted on chitosan–collagen films with the immobilized QHREDGS peptide. The clinical relevance was demonstrated further by assessing the chitosan–collagen hydrogel with immobilized QHREDGS in full-thickness excisional wounds in a db/db diabetic mouse model; QHREDGS showed significantly accelerated and enhanced wound closure compared with a clinically approved collagen wound dressing, peptide-free hydrogel, or blank wound controls. The accelerated wound closure resulted primarily from faster re-epithelialization and increased formation of granulation tissue. There were no observable differences in blood vessel density or size within the wound; however, the total number of blood vessels was greater in the peptide-hydrogel–treated wounds. Together, these findings indicate that QHREDGS is a promising candidate for wound-healing interventions that enhance re-epithelialization and the formation of granulation tissue.

In this paper, the preparation of new supported ionic liquids (SILs) composed of the N-methylimidazolium cation and the quinoline cation is described. They have been confirmed and evaluated by infrared spectroscopy, elemental analysis and thermogravimetric analysis. Six kinds of different SILs included SiO(2)·Im(+)·Cl(-), SiO(2)·Im(+)·BF(4)(-), SiO(2)·Im(+)·PF(6)(-), SiO(2)·Qu(+)·Cl(-), SiO(2)·Qu(+)·BF(4)(-) and SiO(2)·Qu(+)·PF(6)(-). The adsorption characteristics of ferulic acid (FA), caffeic acid (CA) and salicylic acid (SA) on SILs were investigated by static adsorption experiments. It was found that SiO(2)·Qu(+)·Cl(-) had excellent adsorption and desorption capacity to three tested phenolic compounds. The dynamic adsorption characteristics of FA, CA and SA on SiO(2)·Qu(+)·Cl(-) were also studied. The saturated adsorption capacity of FA, CA and SA using SiO(2)·Qu(+)·Cl(-) as adsorbent was 64.6 mg/g, 53.2 mg/g and 72.2 mg/g respectively. Using 70% ethanol as eluent, the saturated desorption efficiencies of FA, CA and SA were 97.2%, 90.3% and 96.5% respectively. Thus, SiO(2)·Qu(+)·Cl(-) had strong adsorption and separation capacity for FA, CA and SA.

Background: Phenolic constituents were the principle bioactivity compounds exist in Gynura divaricata, little phenolic compounds were reported from the plant previously. Materials and Methods: 60% ethanol extract from the leaves of Gynura divaricata were isolated and purified by column chromatography of Silica gel, ODS and Sephadex LH-20, the structures of the isolated compounds were identified by UV, 1H-NMR, 13C-NMR and MS spectroscopic techniques. Additionally, a high-performance liquid chromatography-diode array detector-electrospray ionization-mass (HPLC-DAD-ESI-MS) analytical method was developed to identify some minor constituents in the n-butanol fraction of the ethanol extract of Gynura divaricata. Results: Six flavonols and one Dicaffeoylquinic acid were isolated from the leaves of Gynura divaricata, and these compounds were identified as follows: quercetin (1), kaempferol (2), kaempferol-3-O-β-D-glucopyranoside (3), quercetin-3-O-rutinoside (4), kaempferol-3,7-di-O-β-D-glucopyranoside (5), kaempferol-3-O-rutinoside-7-O-β-D-glucopyranoside (6), and 3,5-dicaffeoylquinic acid (7). A total of 13 compounds, including 9 flavonol glycosides and 4 phenolic acids, were tentatively identified by comparing their retention time (RT), UV, and MS spectrum values with those that had been identified and the published data. Conclusion: This was the first time to use the HPLC-DAD-ESI-MS method to identify the phytochemicals of the genera Gynura. Moreover, compounds (6) and (7) have been isolated for the first time from the genus Gynura.

Optimization of total flavonoid compound (TFC) extraction from Gynura medica leaf was investigated using response surface methodology (RSM) in this paper. The conditions investigated were 30-60% (v/v) ethanol concentration (X(1)), 85-95 °C extraction temperature (X(2)) and 30-50 (v/w) liquid-to-solid ratio (X(3)). Statistical analysis of the experiments indicated that temperature and liquid-to-solid ratio significantly affected TFC extraction (p0.01). The Box-Behnken experiment design showed that polynomial regression models were in good agreement with the experimental results, with the coefficients of determination of 0.9325 for TFC yield. The optimal conditions for maximum TFC yield were 55% ethanol, 92 °C and 50 (v/w) liquid-to-solid ratio with a 30 min extraction time. Extracts from these conditions showed a moderate antioxidant value of 54.78 μmol quercetin/g dry material (DM), 137.3 μmol trolox/g DM for 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 108.21 μmol quercetin/g DM, 242.31 μmol trolox/g DM for 2,2-azino-bis-(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS(+)), respectively. HPLC-DAD-MS analysis showed that kaempferol-3-O-glucoside was the principal flavonoid compound in Gynura medica leaf.

This paper examines the use of the defatted rice bran hemicellulose B (RBHB), and insoluble dietary fibre (RBDF), as two functional ingredients added to bakery products. The results show that the RBHB from defatted rice bran had high water-binding and swelling capacity. RBHB exhibited high fat binding capacity. However, RBHB was found to be low viscous. Addition of 1%, 2% and 3% RBHB preparation reduced loaf volume significantly and increased the firmness of the breads. Sensory evaluations revealed that breads with 1%, 2%, 3% RBHB and 2%, 4% RBDF were overall acceptable. This confirms that the RBHB and RBDF preparation from defatted rice bran has great potential in food applications, especially in development of functional foods including functional bakery products.

Ginkgo biloba contains terpene triclactones of high pharmaceutical value such as ginkgolides. 1-hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate (HMBPP) reductase (HDR) is proved to be the terminal-acting enzyme in the plastid MEP pathway which provides isoprenoid precursors for the biosynthesis of ginkgolides. The full-length cDNA encoding HDR, designated as GbHDR (Genbank Accession Number DQ364231), was isolated for the first time from G. biloba by RACE method. GbHDR contained a 1,422-bp open reading frame encoding 474 amino acids. The deduced GbHDR protein, showing high identity to HDRs of other plant species, was predicted to possess a chloroplast transit peptide at the N-terminal and four conserved cysteine residues. Two-dimensional structural analysis showed that GbHDR had a similar secondary structure with HDR from Arabidopsis thaliana. Southern blot analysis indicated that GbHDR belonged to a small gene family. Transcription pattern analysis revealed that GbHDR had high transcription in roots, and low in leaves and stems. The cloning of GbHDR gene will enable us to further understand the role of GbHDR involved in terpene triclatones biosynthetic pathway in G. biloba at molecular level.

Background Triple-negative breast cancer (TNBC) is associated with an aggressive clinical course due to the lack of therapeutic targets. Therefore, identifying reliable prognostic biomarkers and novel therapeutic targets for patients with TNBC is required. Proline, glutamic acid, leucine rich protein 1 (PELP1) is a novel steroidal receptor co-regulator, functioning as an oncogene and its expression is maintained in estrogen receptor (ER) negative breast cancers. PELP1 has been proposed as a prognostic biomarker in hormone-related cancers, including luminal-type breast cancers, but its significance in TNBC has not been studied. Methods PELP1 immunoreactivity was evaluated using immunohistochemistry in 129 patients with TNBC. Results were correlated with clinicopathological variables including patient’s age, tumor size, lymph node stage, tumor grade, clinical stage, histological type, Ki-67 LI, as well as clinical outcome of the patients, including disease-free survival (DFS) and overall survival (OS). Results PELP1 was localized predominantly in the nuclei of carcinoma cells in TNBC. With the exception of a positive correlation between PELP1 protein expression and lymph node stage (p = 0.027), no significant associations between PELP1 protein expression and other clinicopathological variables, including DFS and OS, were found. However, when PELP1 and Ki-67 LI were grouped together, we found that patients in the PELP1/Ki-67 double high group (n = 48) demonstrated significantly reduced DFS (p = 0.005, log rank test) and OS (p = 0.002, log rank test) than others (n = 81). Multivariable analysis supported PELP1/Ki-67 double high expression as an independent prognostic factor in patients with TNBC, with an adjusted hazard ratio of 2.020 for recurrence (95 % CL, 1.022–3.990; p = 0.043) and of 2.380 for death (95 % CL, 1.138–4.978; p = 0.021). Conclusions We found that evaluating both PELP1 and Ki-67 expression in TNBC could enhance the prognostic sensitivity of the two biomarkers. Therefore, we propose that PELP1/Ki-67 double high expression in tumors is an independent prognostic factor for predicting a poor outcome for patients with TNBC.

A new gene, designated as BnPrx (GenBank Accession No. DQ078754), was isolated from oilseed rape (Brassica napus) by SMART Rapid Amplification of cDNA Ends (RACE). The full-length cDNA is 1307 bp long and contains a 1062 bp open reading frame (ORF), which encodes a 354 amino acid peroxidase precursor, with a 31 aa N-terminal signal peptide and a 15 aa C-terminal propeptide. The putative protein has a molecular weight of 38.86 kDa and a calculated pI of 5.85. BnPrx shares high identity with HRPC (89%). BnPrx possesses all active residues and two Ca2+ sites present in Horseradish peroxidase isoenzymes C (HRPC) as well as six N-glycosylation sites. The predicted 3-D structure of BnPrx is very similar to that of HRPC. Assisted by genomic walking technology, the genomic DNA of BnPrx was also cloned, consisting of 3 introns and 4 exons. Thirty-two TATA boxes, 18 CAAT boxes and many cis-elements, such as WUN, MeJR, were found in its promoter region. Southern blot analysis indicated that BnPrx belonged to a small gene family. Northern blot analysis revealed that BnPrx was constitutively expressed in all tested tissues, including roots, stems and leaves, with the high expression in leaves and stems. The expression of BnPrx could be induced by methyl jasmonate (MeJA), salicylic acid (SA), cold and H2O2. The cloning and characterizing of BnPrx might not only help us understand the physiological function and molecular evolution of the large peroxidase gene family more comprehensively, but also provide an alternative way of seeking a more effective and economical substitute for HRPC.

In this paper, the antioxidative and anticancer activities of tea glycoprotein (TGP) in green tea were studied. TGP was extracted from coarse old green tea and purified by Sephadex G-100 gel filtration, and its purity was determined by high performance gel permeation chromatography (HPGPC). The antioxidative activity of the TGP was evaluated by determining the change of the values of the heat-induced oxidation in a linoleic acid system with β-carotene, the decoloration of the 1,1-diphenyl-2-picrylhydrazyl (DPPH), and the superoxide generated from autoxidation of pyrogallol. The results show that the TGP possesses distinctive antioxidative activity. Furthermore, the anticancer activity of the TGP at different concentrations was evaluated by the MTT assay using two kinds of colon cancer cell lines (HCT-15, Caco-2). Dose-dependently TGP exhibited good antiproliferation activity to HCT-15, whereas exhibited very weak antiproliferation activity to Caco-2. Only at a very high concentration (409.6 μmol L−1), the TGP obviously inhibited the proliferation of Caco-2. Whether there is a correlation between the antioxidative and the anticancer activity of the TGP, should be studied further.

Three structurally similar aromatic heterocyclic compounds 2-thiophenecarboxaldehyde (a), 2-furaldehyde (b), 2-pyrrolecarboxaldehyde (c) were chosen and a series of their thiosemicarbazone derivatives(1a-3a, 1b-3b and 1c-3c) were synthesized to evaluate their biological activities as mushroom tyrosinase inhibitors. The inhibitory effects of these compounds on tyrosinase were investigated by using spectrofluorimetry, (1)H NMR titration and molecular docking techniques. From the results of fluorescence spectrum and (1)H NMR titration, it was found that forming complexes between the sulfur atom from thiourea and copper ion of enzyme center may play a key role for inhibition activity. Moreover, investigation of (1)H NMR spectra further revealed that formation of hydrogen bond between inhibitor and enzyme may be helpful to above complexes formation. The results were well coincident with the suggestion of molecular docking and obviously showed that 2-thiophone N(4)-thiosemicarbazone (1a), 2-furfuran N(4)-thiosemicarbazone (1b) and 2-pyrrole N(4)-thiosemicarbazone (1c) are potential inhibitors which deserves further investigation.

Objective To study the chemical constituents of Daphne odora var. margirmta. Method The compounds were isolated and purified by column chromatography of silica gel, ODS, Sephadex LH-20, and their structures were identified by UV, 1H-NMR, 13C-NMR, MS and CD spectroscopic methods. Result Ten compounds were elucidated as wikatrols B (1), kaempferol (2), daphnodorin B (3), daphnodorin D2 (4), daphnodorin A (5), daphnodorin C (6), daphnodorin D1 (7), daphnodorin I (8), daphnetin (9), daphnoretin (10). Conclusion Compounds 1,2,4 were obtained from this plant for the first time,and compound 1 was isolated firstly from the genus Daphne.

Microglial activation and adaptive immunity have been implicated in the neurodegenerative processes in Parkinson disease. It has been proposed that these responses may be triggered by modified forms of alpha-synuclein (alpha-SYN), particularly nitrated species, which are released as a consequence of dopaminergic neurodegeneration. To examine the relationship between alpha-SYN, microglial activation, and adaptive immunity, we used a mouse model of Parkinson disease in which human alpha-SYN is overexpressed by a recombinant adeno-associated virus vector, serotype 2 (AAV2-SYN); this overexpression leads to slow degeneration of dopaminergic neurons. Microglial activation and components of the adaptive immune response were assessed using immunohistochemistry; quantitative polymerase chain reaction was used to examine cytokine expression. Four weeks after injection, there was a marked increase in CD68-positive microglia and greater infiltration of B and T lymphocytes in the substantia nigra pars compacta of the AAV2-SYN group than in controls. At 12 weeks, CD68 staining declined, but B- and T-cell infiltration persisted. Expression of proinflammatory cytokines was enhanced, whereas markers of alternative activation (i.e. arginase I and interleukins 4 and 13) were not altered. Increased immunoreactivity for mouse immunoglobulin was detected at all time points in the AAV2-SYN animals. These data show that overexpression of alpha-SYN alone, in the absence of overt neurodegeneration, is sufficient to trigger neuroinflammation with both microglial activation and stimulation of adaptive immunity.

Background The protein alpha-synuclein (α-SYN), which is found in the Lewy bodies of dopamine-producing (DA) neurons in the substantia nigra (SN), has an important role in the pathogenesis of Parkinson’s disease (PD). Previous studies have shown that neuroinflammation plays a key role in PD pathogenesis. In an AAV-synuclein mouse model of PD, we have found that over-abundance of α-SYN triggers the expression of NF-κB p65, and leads to microglial activation and DA neurodegeneration. We also have observed that Fcγ receptors (FcγR), proteins present on the surface of microglia that bind immunoglobulin G (IgG) and other ligands, are key modulators of α-SYN-induced neurodegeneration. Methods In order to study the role of FcγRs in the interactions of α-SYN and microglia, we treated the primary microglial cultures from wild-type (WT) and FcγR−/− mice with aggregated human α-SYN in vitro. Results Using immunocytochemistry, we found that α-SYN was taken up by both WT and FcγR−/− microglia, however, their patterns of internalization were different, with aggregation in autophagosomes in WT cells and more diffuse localization in FcγR−/− microglia. In WT microglia, α-SYN induced the nuclear accumulation of NF-κB p65 protein and downstream chemokine expression while in FcγR−/− mouse microglia, α-SYN failed to trigger the enhancement of nuclear NF-κB p65, and the pro-inflammatory signaling was reduced. Conclusions Our results suggest that α-SYN can interact directly with microglia and can be internalized and trafficked to autophagosomes. FcγRs mediate this interaction, and in the absence of the gamma chain, there is altered intracellular trafficking and attenuation of pro-inflammatory NF-κB signaling. Therefore, blocking either FcγR signaling or downstream NF-κB activation may be viable therapeutic strategies in PD.

Background: Extraction temperature influences the total phenolic content (TPC), total flavonoid content (TFC) of medicinal plant extracts to a great extend. TPC and TFC are the principle activity constituents present in the plant. The effects of extraction temperature on TPC, TFC and free radical-scavenging capacity of Gynura divaricata leaf extracts are worth to study. Materials and Methods: Folin–Ciocalteu and aluminum chloride colorimetric assay were used to determine the TPC and TFC of Gynura divaricata leaf extracts at different temperatures. The antioxidant and free radical-scavenging activity were measured by 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid (ABTS) and phosphomolybdenum methods. Results: TPC and TFC were significantly elevated with increasing extraction temperature (from 40°C to 100°C). However, TPC and TFC were not significantly different (P > 0.05) at the extraction temperatures 90°C and 100°C. Also, the extracts obtained at a higher temperature exhibited a significant free radical-scavenging activity compared with extraction at lower temperatures (P < 0.05). The TPCs (13.95-36.68 mg gallic acid equivalent/g dry material) were highly correlated with DPPH (R2 = 0.9229), ABTS (R2 = 0.9951) free radical-scavenging capacity, and total antioxidant activity (R2 = 0.9872) evaluated by phosphomolybdenum method. Conclusion: The TPC and TFC of G. divaricata leaf was significantly influenced by the extraction temperatures, which were the main antioxidant constituents present in the G. divaricata plant.

Overexpression of alpha-synuclein (α-SYN), a protein which plays an important role in the pathogenesis of Parkinson's disease (PD), triggers microglial activation and adaptive immune responses, and leads to neurodegeneration of dopaminergic (DA) neurons. We hypothesized a link between the humoral adaptive immune response and microglial activation in α-SYN induced neurodegeneration. To test this hypothesis, we employed adeno-associated virus serotype 2 (AAV2) to selectively over-express human α-SYN in the substantia nigra (SN) of wild-type mice and FcγR-/- mice, which lack high-affinity receptors for IgG. We found that in wild-type mice, α-SYN induced the expression of NF-κB p65 and pro-inflammatory molecules. In FcγR-/- mice, NF-κB activation was blocked and pro-inflammatory signaling was reduced. Microglial activation was examined using immunohistochemistry for gp91PHOX. At four weeks, microglia were strongly activated in wild-type mice, while microglial activation was attenuated in FcγR-/- mice. Dopaminergic neurodegeneration was examined using immunohistochemistry for tyrosine hydroxylase (TH) and unbiased stereology. α-SYN overexpression led to the appearance of dysmorphic neurites, and a loss of DA neurons in the SN in wild-type animals, while FcγR-/- mice did not exhibit neuritic change and were protected from α-SYN-induced neurodegeneration 24 weeks after injection. Our results suggest that the humoral adaptive immune response triggered by excess α-SYN plays a causative role in microglial activation through IgG-FcγR interaction. This involves NF-κB signaling, and leads to DA neurodegeneration. Therefore, blocking either FcγR signaling or specific intracellular signal transduction events downstream of FcγR-IgG interaction, such as NF-κB activation, may be viable therapeutic strategies in PD.