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Efficient extraction of the total polyphenols and triterpenes from Celtis leaves with multiple DESs by RSM.

The aim of this study is to explore the possibility of modeling Gitelman’s disease (GIT) with human-induced pluripotent stem cell (hiPSC)-derived kidney organoids and to test whether gene correction using CRISPR/Cas9 can rescue the disease phenotype of GIT. To model GIT, we used the hiPSC line CMCi002 (CMC-GIT-001), generated using PBMCs from GIT patients with SLC12A3 gene mutation. Using the CRISPR-Cas9 system, we corrected CMC-GIT-001 mutations and hence generated CMC-GIT-001corr. Both hiPSCs were differentiated into kidney organoids, and we analyzed the GIT phenotype. The number of matured kidney organoids from the CMC-GIT-001corr group was significantly higher, 3.3-fold, than that of the CMC-GIT-001 group (12.2 ± 0.7/cm2 vs. 3.7 ± 0.2/cm2, p < 0.05). In qRT-PCR, performed using harvested kidney organoids, relative sodium chloride cotransporter (NCCT) mRNA levels (normalized to each iPSC) were increased in the CMC-GIT-001corr group compared with the CMC-GIT-001 group (4.1 ± 0.8 vs. 2.5 ± 0.2, p < 0.05). Consistently, immunoblot analysis revealed increased levels of NCCT protein, in addition to other tubular proteins markers, such as LTL and ECAD, in the CMC-GIT-001corr group compared to the CMC-GIT-001 group. Furthermore, we found that increased immunoreactivity of NCCT in the CMC-GIT-001corr group was colocalized with ECAD (a distal tubule marker) using confocal microscopy. Kidney organoids from GIT patient-derived iPSC recapitulated the Gitelman’s disease phenotype, and correction of SLC12A3 mutation utilizing CRISPR-Cas9 technology provided therapeutic insight.

Objectives To explore the possibility of kidney organoids generated using patient derived human induced pluripotent stem cells (hiPSC) for modeling of Fabry disease nephropathy (FDN). Methods First, we generated hiPSC line using peripheral blood mononuclear cells (PBMCs) from two male FD-patients with different types of GLA mutation: a classic type mutation (CMC-Fb-001) and a non-classic type (CMC-Fb-003) mutation. Second, we generated kidney organoids using wild-type (WT) hiPSC (WTC-11) and mutant hiPSCs (CMC-Fb-001 and CMC-Fb-003). We then compared alpha-galactosidase A (α-GalA) activity, deposition of globotriaosylceremide (Gb-3), and zebra body formation under electromicroscopy (EM). Results Both FD patients derived hiPSCs had the same mutations as those detected in PBMCs of patients, showing typical pluripotency markers, normal karyotyping, and successful tri-lineage differentiation. Kidney organoids generated using WT-hiPSC and both FD patients derived hiPSCs expressed typical nephron markers without structural deformity. Activity of α-GalA was decreased and deposition of Gb-3 was increased in FD patients derived hiPSCs and kidney organoids in comparison with WT, with such changes being far more significant in CMC-Fb-001 than in CMC-Fb-003. In EM finding, multi-lammelated inclusion body was detected in both CMC-Fb-001 and CMC-Fb-003 kidney organoids, but not in WT. Conclusions Kidney organoids generated using hiPSCs from male FD patients might recapitulate the disease phenotype and represent the severity of FD according to the GLA mutation type.

In recent years, deep eutectic solvent (DES) has attracted comprehensive attention on the extraction of natural products, and is regarded as an alternative to traditional organic solvents for the environmental advantages. Twenty-six DESs were compared for their extraction yield of total flavonoids and isovitexin (the main flavonoid in Celtis sinensis) from Celtis sinensis. The results show that the extraction yields of total flavonoids by betaine/glycolic acid (DES8), ethylamine hydrochloride/1,2-propanediol (DES12) and tetrapropylammonium bromide/lactic acid (DES17) are the highest, while the extraction yields of isovitexin by ethylene glycol/malonic acid (DES23), ethylene glycol/glycolic acid (DES24) and 1,2-propanediol/glycolic acid (DES26) are the highest. The extraction conditions using the above six DESs were further optimized systematically. Under optimum conditions, the extraction rates of total flavonoids and isovitexin can be increased up to 95.39 and 10.58 mg g−1, respectively, which were significantly higher than that of methanol extraction. In order to exclude the effect of DESs on the bioactivity of Celtis sinensis extract, the macroporous resin D-101 was used to purify the total flavonoids from DESs extract, and the recovery rates of flavonoids from the above six kinds of DESs were all over 80%. Next, the anti-inflammatory activity of DES extracts was compared using a lymphocyte transformation experiment. The result showed that the inhibition rate of the DES24 extract on the proliferation of Con A-activated T cells was up to 72% with an IC50 value of 124.8 μg mL−1. None of the DESs extracted exhibited cytotoxicity on normal T cells. The mechanism of the anti-inflammatory activity against Con A-activated T cells may be that DES24 flavonoids extract induced the apoptosis of inflammatory T cells, and activated the expression of pro-apoptotic protein. Taken together, DES has showed significant advantages on the extraction of natural products for the relatively mild extraction condition, high yield and environmental-friendliness.

Notoginsenoside R1 and ginsenoside Rg1 are the main active ingredients of

As a fast-growing tree, poplar is widely planted and typically used for wood processing in China. During poplar wood processing, a large amount of poplar sawdust (PS) and poplar leaves (PL) are produced and abandoned. To make full use of poplar resources and clarify the use of poplar as a feed additive, the active ingredients in PS and PL were extracted and isolated, and the anti-inflammatory effects of the extracts on mice with dextran sulfate sodium (DSS)-induced colitis were investigated. In vitro anti-inflammatory experiments showed that the ethyl acetate extract of PS and PL (PSE and PLE, respectively) could significantly inhibit the proliferation of concanavalin A (Con A)-activated lymphocytes. Salicortin, tremulacin and salireposide were identified in both PSE and PLE. Oral administration of PSE and PLE rescued DSS-induced colonic shortening, repaired tissue damage, and decreased the disease activity index (DAI). The antioxidant capacity, including the increased activities of glutathione peroxidase (GSH-Px), total superoxide dismutase (T-SOD and catalase (CAT) and decreased activity of myeloperoxidase (MPO), in the colons of mice with colitis was enhanced through the activation of ERK after treatment with PSE and PLE. The ratio of Th1 to Th17 cells, which can lead to inflammation in the spleen, was significantly decreased by the administration of PSE and PLE, while the phosphorylation of related transcription factors (p65, STAT1, and STAT3) was inhibited. Furthermore, PSE and PLE could induce apoptosis in Con A-activated lymphocytes, which may be associated with the increase in p-TBK1, as the molecular docking results also indicated that salireposide in PSE and PLE could interact with the TBK1 protein. Overall, our study provides a promising feed additive for improving intestinal inflammation in animals and a method for the full utilization of poplar resources.

Background T lymphocytes play an important role in contact hypersensitivity. This study aims to explore the immunosuppressive activity of SBF-1, an analog of saponin OSW-1, against T lymphocytes in vitro and in vivo. Methods Proliferation of T lymphocytes from lymph nodes of mice was determined by MTT assay. Flow cytometry analysis was performed to assess T cell activation and apoptosis. Levels of cytokines were determined by PCR and ELISA. BALB/c mice were sensitized and challenged with picryl chloride and thickness of left and right ears were measured. Results SBF-1 effectively inhibited T lymphocytes proliferation induced by concanavalin A (Con A) or anti-CD3 plus anti-CD28 at a very low dose (10 nM) but exhibited little toxicity in non-activated T lymphocytes at concentrations up to 10 μM. In addition, SBF-1 inhibited the expression of CD25 and CD69, as well as he phosphorylation of AKT in Con A-activated T cells. SBF-1 also induced apoptosis of activated T cells. In addition, SBF-1 also downregulated the induction of the T cell cytokines, IL-2 and IFN-γ in a dose-dependent manner. Furthermore, SBF-1 significantly suppressed ear swelling and inflammation in a mouse model of picryl chloride-induced contact hypersensitivity. Conclusions Our findings suggest that SBF-1 has an unique immunosuppressive activity both in vitro and in vivo mainly through inhibiting T cell proliferation and activation. Its mechanism appears to be related to the blockage of AKT signaling pathway.

Isoorientin and isovitexin, kinds of flavone C-glycosides, exhibit a number of biological properties. In this work, The C-glucosyltransferase (Gt6CGT) gene from Gentiana triflora was cloned and expressed in Escherichia coli BL21(DE3). The optimal activity of Gt6CGT was at pH 7.5 and 50° C. The enzyme was stable over pH range of 6.5–9.0, and had a 1-h half-life at 50° C. The Vmax for luteolin and apigenin was 21.1 nmol/min/mg and 31.7 nmol/min/mg, while the Km was 0.21 mM and 0.22 mM, respectively. Then, we developed an environmentally safe and efficient method for isoorientin and isovitexin production using the coupled catalysis of Gt6CGT and Glycine max sucrose synthase (GmSUS). By optimizing coupled reaction conditions, the titer of isoorientin and isovitexin reached 3820 mg/L with a corresponding molar conversion of 94.7% and 3772 mg/L with a corresponding molar conversion of 97.1%, respectively. The maximum number of UDP-glucose regeneration cycles (RCmax) reached 28.4 for isoorientin and 29.1 for isovitexin. The coupled catalysis reported herein represents a promising method to meet industrial requirements for large-scale isoorientin and isovitexin production in the future.

In recent years, deep eutectic solvent (DES) has attracted comprehensive attention on the extraction of natural products, and is regarded as an alternative to traditional organic solvents for the environmental advantages. Twenty-six DESs were compared for their extraction yield of total flavonoids and isovitexin (the main flavonoid in

Ginkgo acids (GAs) in ginkgo products usually lead to allergies or liver toxicity. In this study, the GA-induced toxicity was attenuated and Con A-induced T-lymphocyte proliferation was inhibited by extracts of Celtis sinensis leaves (ECSL). So, the active ingredients in ECSL were studied to solve the problems caused by GAs. First, the eight components of MeOH extracts were determined by HPLC-DAD/LC-MS. Then, the 12 active ingredients were separated based on the anti-inflammatory activity. Lymphocyte conversion showed that the inhibition rates of apigenin, quercetin, and isovitexin at 100 μM on Con A-activated proliferation of T cells were up to 82.46%, 62.86% and 42.76%, respectively. The inhibition rate on the LPS-induced NO release in RAW 264.7 cells of quercetin, apigenin, isovitexin, and vitexin were exceeding 80% at 100 μM. Taken together, the material foundation for the screen of GAs toxicity-attenuated ingredients were provided here. PRACTICAL APPLICATIONS: Ginkgo biloba extracts (EGBs) have been conducted to develop functional food which could increase blood circulation and enhance memory. Nevertheless, people in consumption of ginkgo products, often caused severe allergic reactions due to the potential allergens identified ginkgolic acids (GAs) of ginkgo products. We first find that the extracts of Celtis sinensis leaves can reduce GAs-induced damage on HepG2 liver cells. Then, the bioactive compounds in C. sinensis leaves were separated and purified based on anti-inflammatory activities against T cells. Quercetin, apigenin, and isovitexin showed well anti-inflammatory activities against Con A-activated T-lymphocytes and LPS activated RAW 264.7 macrophages. However, quercetin and apigenin are flavones O-glycosides which are rich in Ginkgo biloba. To solve the problems in Ginkgo biloba products caused by GAs, flavone C-glycoside (isovitexin) may be used for the further study in GAs toxicity-reduction.

Quercetin glycoside derivatives (QGDs) are a class of common compounds with a wide range of biological activities, such as antitumor activities. However, their molecular targets associated with biological activities have not been investigated. In this study, four common QGDs with mutual bioconversion were selected, and studied in the large-scale reverse docking experiments. Network pharmacology analysis showed that most of the four QGDs can bind several potential protein targets that were closely related to breast cancer disease. Among them, a druggable protein, transforming growth factor beta receptor I (TGFBR1/ALK5) was screened via high docking scores for the four QGDs. This protein has been proven to be an important target for the treatment of breast cancer by regulating the proliferation and migration of cancer cells in the past. Subsequently, the molecular dynamics (MD) simulation and MM/GBSA calculation demonstrated that all QGDs could thermodynamically bind with TGFBR1, indicating that TGFBR1 might be one of the potential protein targets of QGDs. Finally, the cytotoxicity test and wound-healing migration assay displayed that isoquercetin, which can perform best in MD experiment, might be a promising agent in the treatment of breast cancer metastasis.

A series of four multifunctional A2B type H3corroles with meso-borneol substituents at the B position have been synthesized and characterized. A detailed analysis of the optical properties was carried out, and a comparison was made with theoretical calculations to identify the key trends in the structure-property relationships. The meso-borneol substituent couples with the corrole core leading to clear CD signals in the B band region. Enhanced anti-cancer properties are observed in vitro relative to (−)-borneol with cell inhibition rates of up to 94%.

Ginkgo tea is a kind of health food produced from Ginkgo biloba leaves. The market of Ginkgo tea encountered many difficulties because of its bad palatability and vague function statement. In this study, two kinds of glycosidase were used to improve the flavor of Ginkgo tea, and three kinds of bioactivities were selected to investigate the health care function of the tea infusion. The aroma components extracted by headspace absorb (HSA) method during the making of Ginkgo tea were analyzed by GC-MS. The flavonoids and ginkgolides released into the tea infusion were studied by HPLC. A combination of &beta, glucosidase (&beta, G) and &alpha, rhamnosidase (&alpha, R) was applied during the making of the tea. The contents of characteristic aroma components and the release of total flavonoids and ginkgolides were increased significantly by adding &beta, G and &alpha, R. The composition of flavone glycosides was changed greatly. The free radical scavenging, inhibition of inflammatory cell activation, and tumor cytotoxicity activities of the tea were demonstrably improved. According to the release of active components, Ginkgo tea can be brewed repeatedly for at least three times. The enzymes used here show potential application prospects in the making of Ginkgo tea or tea drink to get higher contents of flavonoids, ginkgolides, and aroma components.

In this study, a recombinant carotenoid cleavage dioxygenase 4 was produced from Osmanthus fragrans by E . coli under different bacterial growth conditions and used to develop a biotechnological method for preparation of natural β-ionone from β-carotene. β-ionone was analyzed by HPLC and OfCCD4 activity was measured based on concentration change of β-ionone. At pH 8.0 and 35 °C, the greatest activity of the purified recombinant protein was 14.3 U/mg and the maximum concentration of β-ionone was 71.186 mg/L within 1 h. Both the enzyme activity and the concentration of β-ionone could increase by nearly 6 times with addition of 9% Triton X-100 and 2% liquid paraffin.

Astragalin (kaempferol 3-O-glucoside) is used as a standard to assess the quality of Radix astragali and has exhibited a number of biological properties. In this work, we screened several UDP-dependent glycosyltransferases (UGT) for their potential as efficient biocatalysts for astragalin synthesis. The highest astragalin production with 285 mg/L was detected in the recombinant strain expressing UGT from Arabidopis thaliana (AtUGT78D2). To further improve astragalin production, an efficient UDP-glucose synthesis pathway was reconstructed in the recombinant strain by introducing sucrose permease, sucrose phosphorylase, and uridylyltransferase. On the basis of those results, a recombinant strain, BL21-II, was constructed to produce astragalin. By optimizing conversion conditions, astragalin production was increased from 570 to 1708 mg/L. The production was scaled up using a fed-batch fermentation, and maximal astragalin production was 3600 mg/L, with a specific productivity of 150 mg/L/h after 24 h incubation and a corresponding molar conversion of 91.9%, the highest yield reported to date.

2-[[5, 6, 7, 8-Tetrahydro-4-phenyl-7, 7-dimethyl-6, 8-methanoquinazolin-2-imino] methyl] phenol (3) is a novel tetrahydroquinazolin-2-amine-based high selective fluorescent sensor for Zn2+ and was successfully synthesized from nopinone. Its optical properties have been investigated with UV–vis absorption spectra, fluorescence emission spectra and confocal fluorescence imaging, respectively. The sensor could enhance fluorescence after adding some Zn2+ and emitted green light in ethanol. A linear relationship between Zn2+ concentration (Zn2+, 0.5–13×10−6 M) and fluorescence intensity was obtained for determining the content of Zn2+ with a detection limit of 2.35×10−6 mM, y=76.567x+18.469, R2=0.9956. A grinding method was used to detect the presence of Zn2+ in a solid state. Two clear confocal fluorescence images of watermelon peel veins were provided by adding compound 3 into the watermelon peel slices containing Zn2+ ion, and the new method could be used to detect Zn2+ in fruits. Thus, compound 3 synthesized from nopinone is a fluorescent probe and fluorescence imaging agent.

Quinazolin-2-amine derivatives (4–6) were successfully synthesized from β-pinene derivative nopinone and characterized by Fourier transform infrared spectroscope (FT-IR), nuclear magnetic resonance (NMR), mass spectrometry, and X-ray single crystal diffraction. Then, their optical and thermal properties were characterized with ultraviolet–visible spectroscopy, photoluminescence (PL) spectroscopy, thermogravimetric analysis, and confocal fluorescent microscopic imaging. Three target compounds had enhanced fluorescence in solid and solution states and fluorescent quenching occurred when acid or base was added to the solution. Thermal decomposition temperatures of compounds 4–6 were 138, 290, and 291 °C, respectively and no compound was cytotoxic to normal L02 hepatocyte cells, and confocal fluorescent microscopic imaging in human lung A549 adenocarcinoma cells was realized using these compounds. These new fluorescent compounds may be candidates for future fluorescent bio-imaging agents.

A novel dual-responsive fluorescence probe 6 was constructed by introducing a dipicolyamine moiety on the BODIPY-based skeleton via Sonogashira coupling reaction and confirmed by 1H NMR, 13C NMR, elemental analysis and high-resolution mass spectroscopy (HRMS). The present probe 6 exhibited high selectivity and sensitivity toward Cu2+ and Ni2+ detection over other competitive metal ions by fluorescence quenching phenomena, respectively. In addition, the detection limits (DLs) for Cu2+ and Ni2+ were calculated to be as low as both 0.1 μM respectively, which were lower than the maximum allowable level of the World Health Organization (WHO) limit for the drinking water. The stoichiometric ratio of the probe 6 toward the Cu2+ or Ni2+ ions was determined to be 1:1 according to the result of the Job's plot and HRMS analysis. These results indicated that probe 6 can be used as a selective ‘‘turn-off’’ fluorescent sensor for the detection of Cu2+ and Ni2+.

Alcoholic liver disease (ALD) is one of the most common health problems for drinkers, especially in men. Echinacoside (ECH), a natural phenylethanoid glycoside welcomed by the market, has been shown to have a variety of biological activities, such as neuroprotective, anti-fatigue, anti-diabetes and so on. Here, the protective effect and the underlying mechanism of ECH on ethanol-induced liver injuries were studied. In vitro, the HepG2 cells were treated with ECH prior to ethanol. In vivo, C57BL/6 J mice were fed a Lieber-DeCarli ethanol liquid diet and gave with or without 100 mg/kg ECH for 10 days. Our experiments showed that ECH significantly enhanced the levels of antioxidants and reduced the level of ROS, thus attenuating ethanol-induced oxidative stress. Besides, ECH attenuated lipid accumulation caused by ethanol, as evidenced by oil-red O staining, histological examination and the quantification of TG and TC. Finally, ECH increased the level of PPAR-α, and reduced the levels of SREBP-1c and FASN. When PPAR-α inhibitor was introduced in the system, the effects of ECH on SREBP-1c and FASN were reversed. Taken together, our study suggest that ECH can protect against ethanol-induced liver injuries via alleviating oxidative stress and hepatic steatosis by affecting SREBP-1c/FASN pathway via PPAR-α.

A series of five C2 Symmetric A2B2 type borneol-porphyrin hybrids have been synthesized and characterized. Moreover, a detailed analysis of their optical and redox properties was carried out by comparing the results obtained via optical spectroscopy and electrochemistry. The current results demonstrated that the meso borneol-substitutions excitation coupled strongly with porphyrin core that leads to significant CD signals in the Soret band region, and molecular anti-cancer behaviors were also investigated.

Although ginkgolic acids (GAs) are identified as allergenic components in the leaves and seed coat of ginkgo biloba, it is widely revealed that GAs can exhibit various promising biological activities such as anti-tumor activity and antibacterial activity. Thus, investigating molecular targets underlying biological activities of GAs can contribute to the further development and utilization of GAs. In this paper, five kinds of common GAs were prepared for the large-scale reverse docking experiment. The results obtained by network analysis displayed that GAs can directly target at least four important druggable proteins: peroxisome proliferator-activated receptor (PPARA, PPARD, and PPARG), and dihydroorotate dehydrogenase (DHODH). Subsequently, molecular dynamics (MD) simulations and MM/GBSA calculations demonstrated that all the GAs can bind tightly with DHODH thermodynamically, implying that DHODH was probably considered as one of potential protein targets of GAs. Given the clinical significance of DHODH, these bioactive molecules (GAs) could serve as promising therapeutic agents against acute myeloid leukemia (AML) or the starting scaffold for the development of natural DHODH inhibitors.

To comprehend the molecular mechanisms that control the differences in the composition of Osmanthus essential oils, the RNA-seq data and differentially expressed genes in different cultivar Osmanthus were studied. cDNA libraries of "jinqiugui," "baijie," and "rixianggui" were sequenced using Illumina HiSeq TM 2000. All of the enzymes involved in ionone synthesis were verified. DEGs were revealed and their enriched pathways were analyzed. A total of 20 DEGsencoding four enzymes that were potential candidates involved in ionone biosynthesis, as well as ispH, GPPS, ZDS, and CCD. It provided a way for Osmanthus oil monomer material to be synthesized in vitro.

Phosphatase of regenerating liver-3 (PRL-3) has been reported to have a critical role in metastatic progression of cancers. Here, we investigate how PRL-3 increases the malignant degree of melanoma cells. The expression of PRL-3 increased gradually during the malignant progression of melanoma. The phosphorylation of Akt was elevated in highly malignant melanoma cells, which was accompanied by a decrease in nuclear phosphatase and tensin homolog (PTEN). The phosphorylation of NHERF1 in the serine site was regulated by PRL-3 and showed cytoplasmic translocation upon dephosphorylation, which resulted in a decrease in nuclear PTEN. The co-translocation of NHERF1 and PTEN from the nucleus to the cytoplasm was observed during the malignant progression of melanoma cells. Tumor growth was inhibited significantly, and the survival was prolonged upon knockdown of cytoplasmic NHERF1 in B16BL6 cells prior to the inoculation into mice. Taken together, to our knowledge previously unreported, we have identified NHERF1 as a potential substrate of PRL-3. Its phosphorylation status as well as its change in cellular localization and association with PTEN correlated with the malignant progression of melanoma. Our data provide an explanation for how PRL-3 promotes the malignant progression of melanoma, as well as a diagnostic marker or therapeutic target for malignant melanoma.

Kaempferol (kae) and its glycosides are widely distributed in nature and show multiple bioactivities, yet few reports have compared them. In this paper, we report the antitumor, antioxidant and anti-inflammatory activity differences of kae, kae-7-O-glucoside (kae-7-O-glu), kae-3-O-rhamnoside (kae-3-O-rha) and kae-3-O-rutinoside (kae-3-O-rut). Kae showed the highest antiproliferation effect on the human hepatoma cell line HepG2, mouse colon cancer cell line CT26 and mouse melanoma cell line B16F1. Kae also significantly inhibited AKT phosphorylation and cleaved caspase-9, caspase-7, caspase-3 and PARP in HepG2 cells. A kae-induced increase in DPPH and ABTS radical scavenging activity, inhibition of concanavalin A (Con A)-induced activation of T cell proliferation and NO or ROS production in LPS-induced RAW 264.7 macrophage cells were also seen. Kae glycosides were used to produce kae via environment-friendly enzymatic hydrolysis. Kae-7-O-glu and kae-3-O-rut were hydrolyzed to kae by β-glucosidase and/or α-L-rhamnosidase. This paper demonstrates the application of enzymatic catalysis to obtain highly biologically active kae. This work provides a novel and efficient preparation of high-value flavone-related products.

Osmanthus fragrans are well-known for their fragrance, but it is wasteful if to discard O. fragrans flower after extracting their essential oils. In this paper, we found that O. fragrans flower residues were rich in flavonoids. Six flavonoids and one phenylethanoid glycoside were isolated from the ethanol extract of O. fragrans flower residues, identified as quercetin (1), rutin (2), verbascoside (3), genistin (4), kaempferol (5), isorhamnetin (6) and naringin (7). In bioactivity study, kaempferol (IC50 = 1.43 μg/mL) showed the best anti-inflammatory activity. Isorhamnetin, quercetin, kaempferol, verbascoside and rutin (the values of IC50 were 18.30, 11.05, 16.88, 20.21 and 22.76 μg/mL, respectively) showed excellent DPPH free radical scavenging activity. Verbascoside performed relatively well at inhibiting the growth of both CT26 colonic carcinoma cells (IC50 = 46.87 μg/mL) and HepG2 hepatocarcinoma cells (IC50 = 30.58 μg/mL). In addition, quercetin and kaempferol showed strong anti-proliferation activity against HepG2 cells.

Icotinib is a potent and specific epidermal growth factor receptor tyrosine kinase inhibitor. In this study, we reported that icotinib had the antitumor activity on human squamous cell carcinoma cell line A431 in vitro. Meanwhile, adhesion to fibronectin and expression of integrin α3 and β1 were significantly reduced in a dose-dependent manner after the treatment of icotinib. Moreover, icotinib induced cell cycle arrested and affected expression of various cell cycle related proteins in squamous cancer cell line A431, whereas it did not cause apoptosis. Furthermore, icotinib remarkably down-regulated phosphorylation of protein kinase B (AKT) though blocking the interaction between 3-phosphoinositide-dependent protein kinase-1 (PDK1) and AKT in A431 cells. Taken together, it is shown that the small molecular compound, icotinib, has an anti-squamous cell carcinoma activity in vitro and its antitumor mechanism is associated with the blockage of the interaction between PDK1 and AKT. These results provide a novel strategy for anti-squamous cell carcinoma therapy.

A series of Schiff bases derived from 4-methylsalicylic acid (4a–4s) were synthesized, 14 of which (4a–4h, 4j–4l, 4n, 4q, and 4s) were reported for the first time. All the synthesized compounds were evaluated for their immunosuppressive activities for the first time. Among them, compound 4o displayed the most potent biological activity against lymph node cells (IC50 = 1.02 μM for lymph node cells and IC50 = 2.17 μM for PI3Kγ). The results of apoptosis and western-blot assays demonstrated that the immunosuppressive activity of compound 4o might be mediated by the inhibition of PI3K/AKT signaling pathway. Docking simulation was performed to position compound 4o into the PI3Kγ structure active site to determine the probable binding model.

Background Thermally stable α-L-rhamnosidase with cleaving terminal α-L-rhamnose activity has great potential in industrial application. Therefore, it is necessary to find a proper method to improve the thermal stability of α-L-rhamnosidase. Results In this study, addition of sorbitol has been found to increase the thermostability of α-L-rhamnosidase from Aspergillus terreus at temperatures ranging from 65 °C to 80 °C. Half-life and activation free energy with addition of 2.0 M sorbitol at 70 °C were increased by 17.2-fold, 8.2 kJ/mol, respectively. The analyses of the results of fluorescence spectroscopy and CD have indicated that sorbitol helped to protect the tertiary and secondary structure of α-L-rhamnosidase. Moreover, the isoquercitrin yield increased from 60.01 to 96.43% with the addition of 1.5 M of sorbitol at 70 °C. Conclusion Our findings provide an effective approach for enhancing the thermostability of α-L-rhamnosidase from Aspergillus terreus, which makes it a good candidate for industrial processes of isoquercitrin preparation. Electronic supplementary material The online version of this article (doi:10.1186/s12896-017-0342-9) contains supplementary material, which is available to authorized users.

The sequence alignment of Rha and MRha. (DOCX 1901Â kb)