Triazine-cored covalent organic framework for ultrasensitive detection of polybrominated diphenyl ethers from real samples: Experimental and DFT study.

Food and environmental safety issues attributable to the polybrominated diphenyl ethers (PBDEs) are gaining increasing attention, and these urge us to establish a high-performance sample-handling technique. In this study, an outstanding adsorption performance with short adsorption time (10 min) was achieved for PBDEs using a novel synthesized dispersive solid-phase extraction adsorbent, a reticulated covalent organic framework with N/O functional groups (i.e., imine linkage, triazine, and methoxy) (TAPT-DMTA-COF). By conducting sufficient experimentation and theoretical simulation on adsorption mechanism, the halogen bond between electronegative N/O atoms of TAPT-DMTA-COF and the electropositive Br atoms of PBDEs were observed to play a more pivotal role than π-π, C-H…π interactions, and hydrophobic effects. Furthermore, the positive linear relation between calculated adsorption energy and Br content directly clarified that enrichment behavior of PBDEs can be attributed to halogen bonding. These data implied that integrated nanostructure (i.e., N/O functional groups and reticulated architecture) effectively enhanced adsorption capacity. In case of PBDE analysis, this approach achieved excellent results with low limits of detection (0.03-0.13 ng L-1). Finally, the promising potential applications of aforementioned method were verified by spiking water, fish, and milk samples with PBDEs; good PBDEs recoveries were obtained.

Dangguishaoyao-San attenuates LPS-induced neuroinflammation via the TLRs/NF-κB signaling pathway.

INTRODUCTION: Dangguishaoyao-San (DSS) is composed of six traditional Chinese medicines, including Angelica sinensis, Paeoniae radix, Rhizoma Ligusticum, Poria cocos, Rhizoma Atractylodis Macrocephalae, and Rhizoma Alismatis. DSS has been reported to be effective in alleviating the symptoms of Alzheimer's disease (AD). The aim of this study was to investigate the mechanism of action of DSS in vitro using lipopolysaccharide (LPS)-stimulated BV-2 microglia cells. MATERIALS AND METHODS: BV-2 cells were pretreated with 0.58-1.16 mg/mL of DSS for 2 h and then treated with 1 µg/mL LPS for 24 h. Cell viability was determined by an 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The protein expression levels were measured by Western blots. Inflammatory factors were detected by enzyme-linked immunosorbent assays (ELISAs). The mRNA levels of inflammatory factors were analyzed by quantitative real-time PCR (qRT-PCR). RESULTS: DSS treatment at concentrations of 0.58-1.16 mg/mL resulted in no significant cytotoxicity. DSS attenuated the release of pro-inflammatory factors, such as interleukin-1ß (IL-1ß), iNOS and tumor necrosis factor-α (TNF-α) in LPS-induced BV-2 cells. DSS attenuated the mRNA expression of pro-inflammatory cytokines, TLR2, and TLR4 and decreased TLR4 and TLR protein levels as well as the phosphorylation of IκB in LPS-induced BV-2 cells. DSS also down-regulated the nuclear translocation of p65. CONCLUSION: This study demonstrated that DSS has a protective effect on neuroinflammation in LPS-induced BV-2 microglia cells through the TLRs/NF-κB signaling pathway.

[Effect of Herba Scutellariae Barbatae flavonoids in delaying aging of Caenorhabditis elegans and human umbilical vein endothelial cells in vitro].

OBJECTIVE: To explore the effects of Herba Scutellariae Barbatae flavonoids (HF) in delaying aging of Caenorhabditis elegans and human umbilical vein endothelial cells (HUVECs) in vitro. METHODS: The effects of 30 or 50 mg/L of HF on nematode life span, reproductive capacity, oxidative stress, and antioxidant enzyme activity of C. elegans were assessed, and the effects of HF on the expressions of the genes encoding antioxidant enzymes and the aging-related genes were analyzed using real-time RT-PCR in both C. elegans and cultured HUVECs. Results Compared with the blank control group, C. elegans with HF treatment showed significantly improved mean and maximum lifespan with a prolonged mean lifespan under acute heat stress at 35 degrees celsius;. HF treatment did not impair the reproductive capacity or cause significant changes in the offspring number of C. elegans. In addition, HF enhanced SOD and CAT activity and up-regulated the expression of daf-16 and sir-2.1 (SIRT1) genes in C. elegans and HUVECs. CONCLUSIONS: HF may delay aging of C. elegans and enhance their resistance to acute heat stress without damaging their reproductive capacity possibly by up-regulating the activity of antioxidant enzymes and expressions of antioxidant genes. HF also may protect endothelial cells against oxidative damage.

Efficacy and Safety of Duloxetine on Osteoarthritis Knee Pain: A Meta-Analysis of Randomized Controlled Trials.

OBJECTIVES: The aim of this meta-analysis was to evaluate the efficacy and safety of duloxetine for management of osteoarthritis knee (OAK) pain. METHODS: A systematic literature search of articles for management of OAK using duloxetine were performed in PubMed, EBSCO, EMBASE, ScienceDirect, MEDLINE, ClinicalTrials.gov, Google Scholar, and Cochrane Central Register of Controlled Trials from the available date of inception until the latest issue (October 2013). Potentially relevant randomized controlled trials (RCTs) regarding to comparison of efficacy and safety of duloxetine with placebo for managing OAK pain were included. Also, studies with specific data regarding to pain reductions and response rate, Patient Global Impression of Improvement (PGI-I), functional improvement, Western Ontario and McMaster Osteoarthritis Index (WOMAC), adverse events (AEs), treatment-emergent AEs (TEAEs), mortality were included and analyzed, and those with confounding conditions were excluded. Studies were assessed for quality using the Jadad five-point score for RCTs. Finally, a meta-analysis of all RCTs eligible for inclusion criteria was performed using Review Manager 5.1 meta-analysis software. RESULTS: Three RCTs that enrolled 1,011 patients were included in our meta-analysis. There were statistically significant differences between patients taking duloxetine and those taking placebo with regard to the reductions in pain intensity (992 patients, mean difference [MD] = -0.88, 95% confidence interval [CI] -1.11--0.65, P < 0.0001), a moderate improvement in pain intensity (>= 30% response rate; 989 patients, risk ratio [RR] = 1.49, 95% CI 1.31-1.70, P < 0.0001), a substantial improvement in pain intensity (>=50% response rate; 989 patients, RR = 1.69, 95% CI 1.27-2.25, P = 0.0004). Statistically significant differences in PGI-I (976 patients, MD = -0.47, 95% CI -0.63 to -0.30, P < 0.0001) and WOMAC-physical function subscale (977 patients, MD = -4.25, 95% CI -5.82 to -2.68, P < 0.0001) were observed. Similarly, more AEs, TEAEs, and discontinuations for any reason were associated with the use of duloxetine than with placebo (1,011 patients, RR = 2.15, 95% CI 1.48-3.11, P < 0.0001; 1,011 patients, RR = 1.32, 95% CI 1.16-1.49, P < 0.0001; 1,011 patients, RR = 1.43, 95% CI 1.14-1.78, P = 0.002, respectively). However, differences in serious AEs were not significantly statistically different. Moreover, no deaths occurred during these three studies. CONCLUSION: This analysis suggests duloxetine (60/120 mg quaque die (QD)), compared with placebo control, resulted in a greater reduction in pain, improved function and patient-rated impression of improvement, and acceptable adverse effects for the treatment of OAK pain after approximately 10-13 weeks of treatment.

[Comparison of Chemical Components of Essential Oil from Ocimum basilicum var. pilosum Extracted by Supercritical CO2 Fluid and Steam Distillation].

OBJECTIVE: To compare the chemical components of essential oil prepared by steam distillation extraction (SD) and supercritical CO2 fluid extraction (SFE-CO2) from Ocimum basilicum var. pilosum whole plant. METHODS: The essential oil of Ocimum basilicum var. pilosum were extracted by SD and SFE-CO2. The chemical components of essential oil were separated and analyzed by gas chromatography-mass spectrometry( GC-MS). Their relative contents were determined by normalization of peak area. RESULTS: 40 and 42 compounds were detected in the essential oil prepared by SD and SFE-CO2 respectively. 25 compounds were common. CONCLUSION: Thereare significant differences of the chemical components between the Ocimum basilicum var. pilosum essential oil prepared by SD and thatby SFE-CO2. Different methods showed different extraction efficiency with a special compound. It might be a good idea to unite several methods in the modern traditional Chinese medicine industry.

[Comparison of the chemical components of essential oil extracted by supercritical CO2 fluid and steam distillation from Pteris multifida].

OBJECTIVE: To compare the chemical components of essential oil extracted by supercritical CO2 fluid extraction (SFE-CO2) and steam distillation extraction (SD) from Pteris multifida. METHODS: The essential oil of Pteris multifida was extracted by SFE-CO2 and SD. The chemical components of essential oil were separated and analyzed by GC-MS. Their relative contents were determined by normalization of peak area. RESULTS: Twenty -seven compounds in the essential oil extracted by SFE-CO2 and 45 compounds in the essential oil extracted by SD were identified respectively. There were 11 common components. CONCLUSION: The chemical components of essential oil extracted by SFE-CO2 are different from that extracted by SD.

[Content analysis of total polyphenol in the leaves of Jatrophy curcas. L].

OBJECTIVE: To establish a method for analyzing the content of total polyphenols in leaves of Jatropha curcas. L. METHODS: Gallic acid was used as reference substance, the content of total polyphenols was analyzed Folin-Ciocalteu chromatometry. RESULTS: There was a good linearity for gallic acid in the range of 0.002-0.010 g.L(-1). The content of total polyphenols in the leaves of Jatropha curcas. L was approximately 6.74% with RSD 0.75%. The sample solution was stable during 10-90 min with RSD 0.28%. The precision RSD was 0.23% and the average recovery 99.85% (n=5). CONCLUSION: This method is simple, fast and reproducible.

[Gas chromatography-mass spectrometric analysis of Jatropha curcas leaf extracts prepared by supercritical fluid CO2 extraction].

OBJECTIVE: To analyze the bioactive components in Jatropha curcas leaves using gas chromatography-mass spectrometry (GC-MS). METHODS: The bioactive components were extracted from J. curcas leaves by supercritical fluid CO2 extraction and analyzed by using GC-MS. RESULTS: Seventy peaks were detected by GC-MS, and 43 compounds were identified (61.43%). Among the identified compounds, 16 had a content of more than 1%, and the total contents of these 16 compounds reached 81.36%. The four most abundant components were 22,23-dihydro-stigmasterol (16.14%), alpha-tocopherol (15.18%), beta-amylin (7.73%) and dotriacontanol (7.02%). The content of gamma-tocopherol reached 2.88% and vitamin E reached 18.06% in the extract. CONCLUSION: J. curcas leaves contain multiple compounds with anti-tumor, anti-virus and antimicrobial activities.

[Sensitivity of Jatropha curcas seeds to (60)Co-gamma radiation and their medial lethal doses in radiation breeding].

OBJECTIVE: To study the sensitivity of Jatropha curcas seeds from three different locations to (60)Co-gamma radiation and to determine the medial lethal doses (LD50) of (60)Co-gamma radiation for these seeds. METHODS: Six different radiation doses (0, 100, 150, 200, 250 and 300 Gy) were used. Based on the germination rate 50%, LD50 doses of (60)Co-gamma radiation for the seeds were calculated using linear regression equation. RESULTS: LD50 doses of (60)Co-gamma radiation for these seeds were 178 Gy (seeds from Guangdong), 132 Gy (seeds from Hainan) and 198 Gy (seeds from India) respectively. Increasing radiation doses caused more significant changes in leaf shape of the M1 seedlings. CONCLUSION: The results provides an important experimental basis for the radiation breeding of the important herbal and energy plant J. curcas.

[Oil contents and fatty acid composition in Jatropha curcas seeds collected from different regions].

OBJECTIVE: To compare the oil contents and fatty acid composition among the samples of Jatropha curcas L. seeds collected from China (Guangdong, Hainan, and Guizhou Provinces) and India. METHODS: Soxhlet extraction method and gas chromatography-mass spectrometry (GC-MS) were employed to determine the oil contents of Jatropha seeds and the fatty acid composition of Jatropha oil. RESULTS: The seed oil contents (dry basis) were 32.43% (Guangdong), 31.41% (Hainan), 37.56% (Guizhou) and 41.04% (India), respectively. Twelve different fatty acids were detected by GC-MS, and the content of total unsaturated fatty acids accounted for 80.93%, 79.53%, 77.24% and 78.22% of the total fatty acids in the samples collected from Guangdong, Hainan, Guizhou and India, respectively. CONCLUSION: There are differences in the oil contents and fatty acid composition among the J. curcas seeds collected from different regions, and attention should be given to these differences in the introduction and breeding of J. curcas.

[Tumor-specific T cell recptor gene transfection promotes memory T cell differentiation in vitro].

OBJECTIVE: To investigate effect of tumor-specific T cell receptor gene transfection on memory T cell differentiation in vitro. METHODS: TCRVbeta7.1 gene was transferred into peripheral blood mononuclear cells (PBMCs) obtained from healthy adults, and the expression of Vbeta7.1 was detected by flow cytometry before and after the transfection. Memory T cell differentiation was induced by stimulation with the hepatocarcinoma cell line BEL-7402 in vitro. The expression of surface molecules CD45RO, CD45RA and CCR7 was analyzed by flow cytometry to identify the phenotype and subsets of the memory T cells. Fluorescence-activated cell sorting was performed to detect the apoptosis of the tumor cells, and enzyme-linked immunoabsorbent assay was used to determine the production of interferon-gamma (IFN-gamma) for assessing the immune function of the memory T cells. RESULTS: Flow cytometry showed that TCRVbeta7.1 gene was efficiently expressed after transfection. After stimulation by the tumor cells in vitro, the expression of CD45RO in TCRVbeta7.1 gene-modified T cells increased gradually, and analysis of the coexpression of CD45RA and CCR7 revealed that the effector memory T cells constituted the majority of the differentiated memory T cells. The apoptotic rate of the tumor cells induced by the T cells increased significantly with also obviously increased INF-gamma secretion in the memory T cells. CONCLUSION: Tumor-specific TCRVbeta7.1 gene transfection can promote the differentiation of the memory T cells, the majority of which belongs to effector memory T cells that perform immune functions by inducing apoptosis and cytokine secretion.

[Antitumor effect of GPI-CD80 fusion protein in nude mice].

OBJECTIVE: To study the antitumor effect GPI-CD80 fusion protein and its mechanisms. METHODS: A tumor vaccine was prepared by culturing HepG2 cells in the presence of purified GPI-CD80 followed by inactivation with mitomycin, with mitomycin-inactivated HepG2 cells as the control group. The two preparations were co-cultured with nude mouse splenic lymphocytes, and the changes of lymphocyte proliferation and the production of interleukin-2 (IL-2) and interferon-gamma (IFN-gamma) were detected by MTT assay. The cytotoxic T-lymphocyte (CTL) activity was evaluated by LDH-release assay, and the changes of gross tumor volume were measured in tumor-bearing nude mice after administration of different vaccines. RESULTS: The application of GPI-CD80 tumor vaccine resulted in significantly increased optical density, IL-2 and IFN-gamma levels and CTL activity of the nude mouse splenic lymphocytes in comparison with the control groups. The average tumor volume in nude mice treated with GPI-CD80 tumor vaccine was significantly smaller than that in negative control and blank control groups. CONCLUSION: GPI-CD80 fusion protein may inhibit the tumor growth velocity in nude mice, possibly by promoting lymphocyte proliferation, stimulating the production of the cytokines IL-2 and IFN-gamma, and enhancing of CTL activity.

[Efficient and rapid non-test tube cloning of Jatropha curcas].

OBJECTIVE: To develop a new technique for efficient and rapid non-test tube cloning of the medicinal and energy- producing plant Jatropha curcas. METHODS: Using the mini-stem fragment (2-3 cm) of Jatropha curcas with merely one axillary bud as the explant, the effect of an auxin IBA concentration on the plantlet regeneration was studied. RESULTS AND CONCLUSION: When treated with 1 mg/LIBA for 1h, the explants showed the most rapid propagation. The mini-stem fragments high root regeneration ratio (96.7%), short root regeneration period (18.2-/+2.0 d), large number of new roots per explant (6.3-/+1.8), and long total root length (6.8-/+3.5 cm), demonstrating that this technique can be a simple and efficient method for rapid non-test tube cloning of Jatropha curcas of potential industrial value.

[The protective effect of Astragalus membranaceus extraction on cryopreserved primary-cultured human fetal hepatocytes].

OBJECTIVE: To observe the influence of Astragalus membranaceus Extraction (AE) on the primary-cultured human fetal hepatocytes stored in liquid nitrogen and explore a new method for the cryopreservation of human hepatocytes with improved function. METHODS: Human fetal hepatocytes were harvested by two-step collagenase perfusion, and then stored in a liquid nitrogen for one month with five different cryoprotectants (I: 10% DMSO, II: 5% DMSO + 2 mg/L AE, III: 5% DMSO + 20 mg/L AE, IV: 5% DMSO + 60 mg/L AE, V: 5% DMSO + 100 mg/L AE). One month later, the cells were thawed rapidly and the viability, morphology and basic function of them were tested. RESULTS: The human fetal hepatocytes in different groups showed various levels of viability, morphological manifestation and cell function respectively. After thawing, the viability rate and flash adhering rate in group IV and V had no significant difference with group I (P > 0.05), but were higher than group II and III (P < 0.05); the cell function analysis in the group IV, the results of ALB and AST level determination, NH4Cl transformation test, were the best among the groups (P < 0.05). CONCLUSION: AE can provide protection for human fetal hepatocytes in cryopreservation, and the best performance concentration level of its is 60 mg/L; the preservation dosage of DMSO can be reduced when combined with AE in the preservation solution, which shows that AE has a synergistic effect with DMSO.

[Effect of Scutellaria barbata extract against human hepatocellular Hep-G2 cell proliferation and its mechanism].

OBJECTIVE: To observe the effects of Scutellaria barbata extract (ESB) on human hepatoma cell line Hep-G2 proliferation in vitro and explore the mechanism. METHODS: The inhibitory effect of ESB on Hep-G2 proliferation was estimated by MTT assay, and the morphological changes of the cells were observed under optical and electron microscopes. Distribution of cell cycle, cell apoptosis and the protein expressions of apoptosis-associated genes as bcl-2, bax and fas were analyzed using flow cytometry. RESULTS: ESB inhibited the proliferation of Hep-G2 cells in a time- and dose-dependent manner. ESB treatment for 72 h resulted in changes of early apoptotic morphology of the cells as observed under optical and the transmission electron microscopes and increased cell apoptosis. Cell cycle analysis revealed decreased S-phase and increased G0/G1-phase cells. Fas expression was significantly up-regulated in response to ESB treatment whereas Bcl-2 and Bax expressions underwent no significant changes. CONCLUSION: ESB can inhibit Hep-G2 cell proliferation, induce cell cycle block, and increase cell apoptosis, which may relate to the activation of FNFR superfamily.

[Inhibition activity of Scutellariae barbata extracts against human hepatocellular carcinoma cells].

OBJECTIVE: To investigate the inhibition effect of Scutellariae barbata extracts on the proliferation of human hepatocellular carcinoma cell line QGY-7701. METHODS: The inhibition activity of the extracts against cell line QGY-7701 was estimated by MTT assay. Morphologic changes of the cells were observed under light microscope and electronic microscope. Cell cycle distribution, cell apoptosis and expressions of apoptosis-associated genes as Bcl-2, Bax and Fas were analyzed by flow cytometry. RESULTS: Extracts of Scutellariae barbata could inhibit the proliferation of QGY-7701. When treated with the extracts for 72 h, cells at the early stage of apoptosis showed morphologic changes, apoptotic cells increased, cells at G0/G1 phase decreased and those at G2/M phase increased, the expression of Bax significantly up-regulated and that of Bcl-2 down-regulated with no change in Fas gene. CONCLUSION: The extracts of Scutellariae barbata inhibit the proliferation of QGY-7701, which may relate to the activation of anti-oncogene Bcl-2, induction of cell apoptosis and inhibition of G to S phase cell cycle progress.

[Therapeutic effects of koumine on psoriasis: an experimental study in mice].

OBJECTIVE: To study the therapeutic effects of koumine on psoriasis in mouse models. METHODS: The effects of koumine on epithelial cell mitosis and epidermal cell differentiation was evaluated by collecting the samples of the vaginal mucous and squamous epidermis at the tail of mice treated with methotrexate or koumine at different doses. The levels of interleukin (IL)-2 were detected with enzyme-linked immunosorbent assay. RESULTS: High and intermediate doses of koumine showed remarkable inhibitory effect on mouse vaginal epithelial cell mitosis and promoted the formation of epidermal granular layer in the scales at the mouse tail. Three concentrations at 6, 30, 150 mg/kg of koumine decreased serum IL-2 level in the mice. CONCLUSION: The therapeutic effect of koumine against psoriasis is related to the inhibition of epidermal cell proliferation, promoting the formation of granular cells and decreasing the serum level of IL-2.

[Effect of koumine on proliferation of murine CD4+ T cells purified by magnetic-activated cell sorting in vitro].

OBJECTIVE: To assess the separation efficiency of magnetic-activated cell sorting in the purification of CD4+ T cells from murine spleen, and observe the effects of koumine on the proliferation of the separated cells. METHODS: CD4+ T cells were isolated from murine spleen by magnetic-activated cell sorting (MiniMACS). Fluorescence-activated cell sortering was employed to determine the purity of CD4+ T cells before and after the separation procedure followed by evaluation of the cell viability using trypan blue staining. Concanavalin A- (ConA, 5 microg/ml) or phytahematoagglutinin (PHA,1 mg/ml)-induced murine T cells were treated with different concentrations of koumine (10-320 microg/ml), and their proliferation was determined by MTT colorimetry, and enzyme-linked immunosorbent assay was used to measure IL-2 level in the cell culture supernatant. RESULTS: The purity of CD4+ T cells reached (90.3+/-5.8)% after the purification with a cell viability of (94.9+/-3.6)%. Koumine (20-320 microg/ml) dose-dependently inhibited ConA- or PHA-induced proliferation of murine lymphocytes as compared with the controls (P<0.05). Koumine (20, 100, and 200 microg/ml) significantly decreased the level of IL-2 in comparison with the control group (P<0.05). CONCLUSIONS: CD4+ T cells of high purity can be obtained from murine spleen using MiniMACS without impairing the viability of the cells. Koumine significantly inhibits the proliferation of murine CD4+ T cells due to its immunosuppressive effect and inhibition of IL-2 secretion.

[Infrared spectrum and nuclear magnetic resonance analysis of the alkaloid extract of Gelsemium produced in east Guangdong province].

OBJECTIVE: To identify the structure of the alkaloid extract of Gelsemium from east Guangdong province of China. METHODS: Fourier transform infrared (FTIR) absorption spectrometry, hydrogen and carbon spectra of nuclear magnetic resonance (NMR) and distortionless enhancement by polarization transfer (DEPT) analysis were used for the structural identification of the alkaloid exstract of Gelsemium. RESULTS: The frequency, intensity and shape of the extract's characteristic peaks in infrared absorption spectra (4,000.0-400.0 cm(-1)) and (1)H NMR, (13)C NMR were recognized and compared. The molecular structure of the sample was consistent with the theoretically derived model. CONCLUSION: The extract is structurally identical to koumine, which may provide evidence for its safe clinical application and establishment of Chinese medicine fingerprint database of Gelsemium.

[Determination of scopoletin and umbelliferone contents in Saussurea medusa Maxim by high-performance liquid chromatography].

The contents of scopoletin and umbelliferone in Saussurea medusa Maxim were determined by high-performance liquid chromatography (HPLC) using Merck Lichrospher100 RP-18e column (250 mm x 4.0 mm, 5 microm) with the mobile phase of methanol-tetrahydrofuran-water solution and the absorbance of the compounds observed at 346 nm. HPLC yielded an average recovery and RSD of 100.07% and 1.42% for scopoletin, and 99.41% and 2.06% for umbelliferone, respectively, suggesting that HPLC is a simple, rapid and accurate method for quality control of Saussurea medusa Maxim.