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Response surface methodology was employed to model and optimize the electrodialysis process for Reverse osmosis concentrate (ROC) reclamation in coal-fired power plants. Predictive models were deve...

Abstract Zein-based materials were used to remove Trypan blue from water under flow conditions and in batch tests. In flow tests, zein dissolved at pH = 13 was injected in sand columns and subsequently coagulated with CaCl2, to create an adsorbent filter which removed over 99% of Trypan blue. Batch tests were conducted using zein powder, zein dissolved at pH = 13 and coagulated with CaCl2, Fe2Cl3 or citric acid, and zein dissolved in ethanol and then coagulated with water. The highest Trypan blue removal was achieved with zein powder (4000 mg Trypan blue/kg sorbent, as determined through spectrophotometry), followed by zein coagulated with Fe2Cl3 (500 mg Trypan blue/kg sorbent) and with other salts (140 mg Trypan blue/kg sorbent). Differences in the sorption efficiency are attributed to differences in the surface area. The sorption isotherm of Trypan blue onto zein-based sorbents was a Type II isotherm, suggesting physisorption. Desorption of Trypan blue was limited when zein-based coagulated sorbents were immersed in pure water. Trypan blue could be degraded by free laccase in water, as determined through spectrophotometry and electrospray ionization mass spectroscopy (ESI-MS). Trypan blue could also be degraded by laccase when zein-based laccase-containing sorbents were prepared at pH = 10, using Fe2Cl3 as coagulant. Graphic abstract

Two CO2 formation pathways (chlorogenic acid (CGA) degradation and Maillard reaction) during coffee roasting were investigated. CGA is shown not a major contributor to CO2 formation, as heating of this compound under typical roasting conditions did not release a large quantity of CO2. However, heating of a CGA moiety, caffeic acid, resulted in high yield of CO2 (>98%), suggesting that CGA hydrolysis could be the rate limiting step for CO2 formation from CGA. A large amount of CO2 was detected from glycine-sucrose model system under coffee roasting conditions, implying the importance of Maillard reactions in CO2 formation. Further studies on the heating of various components isolated from green coffee beans showed that CO2 was generated from various green coffee components, including water insoluble proteins and polysaccharides. Around 50% of CO2 was formed from thermal reactions of lower molecular weight compounds that represent ∼25% by weight in green coffee.

Artesunate (ART) is a sesquiterpene lactone isolated from the leafy portions of the Chinese herb Artemisia annua. Here, we evaluated the effect of ART on the prostate cancer (PCa) cell lines DU145 and LNCaP and explored its potential mechanisms. ART inhibited the viability and mobility of DU145 and LNCaP cells. Mechanistically, we found that UCA1, one of the most important lncRNAs in malignancies of the urinary system, may be a potential mediator contributing to the tumor suppressor function of ART. First, the UCA1 level was reduced significantly after being exposed to ART. In addition, UCA1 was up-regulated in prostate cancer tissues compared to hyperplastic prostatic tissues, and a higher UCA1 level predicted poor prognosis in PCa patients. Furthermore, reintroduction of UCA1 into PCa cells reversed the effect of ART on apoptosis and metastatic ability. Then we determined that the miR-184/Bcl-2 axis might be the downstream signaling pathway of UCA1 upon ART treatment. UCA1 binds to miR-184 through its seed sequences and may function as a sponge for miR-184.

From its legendary discovery in Abyssinia (today Ethiopia) to becoming one of the most consumed beverages in the world, coffee has captivated the enthusiasts for centuries due to its unique aroma and taste, as well as its effects as a stimulant in enhancing mental performance (e.g., alertness, concentration, attention). This article provides a brief overview on the production and processing of coffee, focusing on the Coffea arabica and Coffea canephora var. Robusta, also known as Arabica and Robusta, respectively. Differences in chemical compositions (e.g., lipid, sucrose, trigonelline, diterpenes, caffeine, chlorogenic acids) of bean variety contribute to desirable/undesirable sensory attributes, as well as the health implications of the final brew products. Roasting of green beans, which is typically carried out at 170–230 °C for 10–15 min, causes the degradations of polysaccharides, sugars, amino acids, chlorogenic acids, and so on. Concomitantly, a myriad of aroma volatiles and complex condensed products are formed, mainly due to Maillard reaction, Strecker degradation and pyrolytic reactions. The effects of roast time–temperature profiles on a number of key physicochemical phenomena are discussed, including changes in microstructural, formation of aroma species, development of color, and generation of CO2 during roasting. Optimal storage conditions and packaging are important in delaying product staling and to mitigate CO2 degassing issues. These aspects, along with other factors that affect the shelf-life of coffee, are discussed. Finally, a brief literature review on the health implications of coffee consumption is presented, highlighting the importance of several bioactive components (e.g., caffeine, chlorogenic acids, melanoidins, trigonelline, acrylamide, and diterpenes).

Hexavalent chromium is one of the main heavy metal pollutants. As the environmental legislation becomes increasingly strict, seeking new technology to treat wastewater containing hexavalent chromium is becoming more and more important. In this research, a novel modified ultrafiltration membrane that could be applied to adsorb and purify water containing hexavalent chromium, was prepared by polyvinylidene fluoride (PVDF) blending with 2-aminobenzothiazole via phase inversion. The membrane performance was characterized by evaluation of the instrument of membrane performance, infrared spectroscopy (FTIR), scanning electron microscope (SEM), and water contact angle measurements. The results showed that the pure water flux of the PVDF/2-aminobenzothiazole modified ultrafiltration membrane was 231.27 L/m2·h, the contact angle was 76.1°, and the adsorption capacity of chromium ion was 157.75 µg/cm2. The PVDF/2-aminobenzothiazole modified ultrafiltration membrane presented better adsorption abilities for chromium ion than that of the traditional PVDF membrane.

CO2 is one of the major gases formed during coffee roasting, which has important implications on coffee's quality and packaging requirements. In this study, the residual CO2 content and CO2 degassing behavior of an Arabica coffee processed using a fluidized bed roaster, as affected by the roasting temperature–time conditions, were investigated. The results showed that positive correlations existed between the degree of roast (expressed as lightness value) and residual CO2, implying that lightness could be used as an indicator of initial CO2 content in roasted coffee. At the same degree of roast, coffee roasted with high-temperature–short-time process had significantly higher CO2 degassing rate than those with low-temperature–long-time process. Moreover, the CO2 releasing rate increased with the degree of roast. The degassing rate of CO2 in ground coffee was highly dependent on the grind size and roasting temperature, but less dependent on the degree of roast. The different degassing behaviors observed between roasted coffee samples were explained on the basis of chemical composition and microstructural differences.

This study describes the preparation and characterisation of MIL-53(Fe) and its effect on the performance of forward osmosis (FO) membranes. The cellulose acetate (CA)/MIL-53(Fe) hybrid membranes were fabricated using the method of phase inversion by dispersing MIL-53(Fe) in a CA casting solution. To improve the selectivity and permeability performance of FO membranes, the effects of MIL-53(Fe) content, casting solution temperature, coagulation bath temperature, and annealing temperature were studied. Results from scanning electron microscopy, membrane porosity, atomic force microscopy, and water contact angle tests of all the membranes show that the structural properties of the CA/MIL-53(Fe) hybrid membranes were optimised. The results of the FO performance tests, including the values of water flux and reverse salt flux, indicate improvement in the selectivity and permeability properties of the CA/MIL-53(Fe) hybrid membranes. Under the conditions of deionised water as the feed solution and 1 M NaCl solution as the draw solution, the water flux and reverse salt flux of the CA/MIL-53(Fe) hybrid membranes reached 34.9 L/(m2·h) and 2.02 g/(m2·h), respectively, in comparison with CA membranes. This study demonstrates that MIL-53(Fe) can improve the desalination performance and structural properties of FO membranes.

AZD6244, an ATP-uncompetitive inhibitor of mitogen-activated protein kinase 1/2 (MEK1/2), has shown activity in several malignant tumours. However, whether AZD6244 has a function in lymphoma cells is not known. We report that AZD6244 treatment represses the growth of Raji and MOLT4 cells by inducing apoptosis and G1-phase arrest. Using miRNAs array and quantitative RT-PCR, miR-92a was downregulated byAZD6244 treatment through the ERK1/2-AP1 signalling pathway. Overexpression of miR-92a abrogated AZD6244-induced apoptosis and G1-phase arrest, indicating that it is involved in the cytotoxicity of AZD6244 in lymphoma cells. A luciferase reporter assay showed that miR-92a directly targetsthe 3'-UTRs of Bim. Overexpression of miR-92a mimics downregulated Bim mRNA and protein expression level, indicating that miR-92a negatively regulates its expression at both levels. Silencing Bim decreases AZD6244-induced apoptosis and G1-phase arrest, suggesting that Bim contributes to the growth arrest. Thus, miR-92a mediates AZD6244-induced cytotoxicity of lymphoma cells by targeting Bim. Downregulation of miR-92a by AZD6244 is mediated by the ERK1/2-AP1 signalling pathway.

Simultaneous perchlorate and nitrate removal from contaminated groundwater in one reactor has been realized with different methods in the past. The usage of biodegradable polymers as biofilm carriers and carbon source is new. Polymer in this paper was designed out of the copolymer of starch and polyvinyl alcohol. Under polluted water with 2 mg/L of perchlorate and 20 mg/L of NO3-N, it was possible to produce completely denitrification only for 5 h and below the detection limit to perchlorate within 9 h. Results indicating a significant impact of liquor pH on the biodegradation of but slight effect on nitrate reduction. Packed-bed reactor filled with polymer granules could remove 2 mg/L perchlorate and 25 mg/L NO3-N completely with influent flow rate of 1.17 mL/min. Morphological observation indicated the developed biofilm coverage on the outer surfaces of the carriers was dense and primarily composed of bacillus and coccus. The microbes in biofilm decomposed polymer, the chink and filament structure on the carrier surface developed, through metabolism and provided carbon source for them by releasing small organic molecules.

In this study, changes in lightness, roast loss, residual CO2, and total volatiles of an Arabica coffee were investigated under isothermal conditions at 220, 230, 240, and 250 °C. The lightness of the roasted coffee, expressed as L* value, followed two-stage processes that could be modeled using pseudo first-order reaction models, giving activation energies of 59.7 and 170.2 kJ/mol for the first and second stages, respectively. Roast loss data also exhibited two-stage behavior, but followed zero-order reaction kinetics, with activation energies of 52.9 and 181.3 kJ/mol for the first and second stage, respectively. The first-to-second stage transition for L* value and roast loss occurred at light-medium roast. Residual CO2 in the coffee beans correlated negatively with L* value below medium-dark roast degree. However, a reversed correlation was observed above dark roast degree. The volatile compounds generated in roasted coffee were highly dependent on roasting temperature and roast degree.

Ribosomal S6 kinases (RSKs) are directly regulated by extracellular signal-regulated kinase (ERK) signaling and are implicated in cell growth, survival, motility and senescence. The present study observed that RSK1 was overexpressed in primary untreated leukemia patient bone marrow samples compared with the expression at the complete remission stage, using reverse transcription-quantitative polymerase chain reaction (RT-qPCR). In addition, a high RSK1 expression (relative expression ≥10) was associated with a significantly shorter overall survival (P=0.038) compared with that in patients with low RSK1 expression (relative expression

Polygalacturonase-inhibiting proteins (PGIPs) are plant cell wall glycoproteins that can inhibit fungal endopolygalacturonases (PGs). The PGIPs directly reduce the aggressive potential of PGs. Here, we isolated and functionally characterized three members of the pepper (Capsicum annuum) PGIP gene family. Each was up-regulated at a different time following stimulation of the pepper leaves by Phytophthora capcisi and abiotic stresses including salicylic acid, methyl jasmonate, abscisic acid, wounding and cold treatment. Purified recombinant proteins individually inhibited activity of PGs produced by Alternaria alternata and Colletotrichum nicotianae, respectively, and virus-induced gene silencing in pepper conferred enhanced susceptibility to P. capsici. Because three PGIP genes acted similarily in conferring resistance to infection by P. capsici, and because individually purified proteins showed consistent inhibition against PG activity of both pathogens, CaPGIP1 was selected for manipulating transgenic tobacco. The crude proteins from transgenic tobacco exhibited distinct enhanced resistance to PG activity of both fungi. Moreover, the transgenic tobacco showed effective resistance to infection and a significant reduction in the number of infection sites, number of lesions and average size of lesions in the leaves. All results suggest that CaPGIPs may be involved in plant defense response and play an important role in a plant's resistance to disease.

In this study, the effects of particle size (222 to 1085μm), packing amount (7.1 to 10.7g), and brewing volume (113 to 226mL) on physiochemical properties of coffee brews, prepared using a commercial single-serve brewer, were investigated. The results show that decreasing particle size increased the extraction yield by about 63% without changing the extraction of acidic and phenolic compounds, implying finer grinds potentially could be used to reduce the use of coffee ground. Increasing packing amount had no effect on the extraction yield, but did increase the concentration ratio of acidic to phenolic compounds, thus changing the flavor profile of the brew. >80% of the soluble solids were extracted within the first 113mL, while further brewing diluted the brew and introduced more bitter and astringent compounds. This study increased the understanding of single-serve brewing process, which is important to optimize brew quality and minimize production cost.

To investigate the mechanisms by which berberine suppressed the proliferation of human multiple myeloma cells.Human U266 multiple myeloma cell line was tested. Cell proliferation, apoptosis, ultramicrostructure and secretion function were examined using Cell Counting Kit-8 (CCK8), flow cytometry (FCM), electron and fluorescence microscopy, as well as ELISA assay. The microRNAs (miRs) and transcription factors in U266 cells were detected using arrays and verified by qRT-PCR. EMSA and luciferase assays were used to verify the p65-dependent transactivation of miR-21 gene.Treatment of U266 cells with berberine (40-160 μmol/L) suppressed cell proliferation and IL-6 secretion in dose- and time-dependent manners. Meanwhile, berberine dose-dependently induced ROS generation, G(2)/M phase arrest and apoptosis in U266 cells, and decreased the levels of miR-21 and Bcl-2. Overexpression of miR-21 counteracted berberine-induced suppression of cell proliferation and IL-6 secretion. In U266 cells treated with berberine (80 μmol/L), the activity of NF-κB was decreased by approximately 50%, followed by significant reduction of miR-21 level. berberine (80-160 μmol/L) increased the level of Set9 (lysine methyltransferase) by more than 2-fold, caused methylation of the RelA subunit, which inhibited NF-κB nuclear translocation and miR-21 transcription. In U266 cells treated with berberine (80 μmol/L), knockdown of Set9 with siRNAs significantly increased NF-κB protein level accompanying with a partial recovery of proliferation.In U266 cells, berberine suppresses NF-κB nuclear translocation via Set9-mediated lysine methylation, leads to decrease in the levels miR21 and Bcl-2, which induces ROS generation and apoptosis.

Microsomal prostaglandin E synthase-1 (mPGES-1), an inducible enzyme that specifically catalyzes the conversion of prostaglandin H2 (PGH2) to prostaglandin E2 (PGE2), has been reported to be over-expressed in a variety of solid tumor cells and tissues, but not in normal tissues. Its association with leukemia, however, has not been fully investigated. Our study revealed, for the first time, that mPGES-1 is over-expressed in human acute myeloid leukemia HL-60 cells. Cytotoxicity assays and flow cytometry showed that MK886, an inhibitor of mPGES-1, inhibits proliferation of HL-60 cells and induces apoptosis in a dose- and time-dependent manner, which may result from down-regulation of mPGES-1 expression and PGE2 synthesis. Evaluation of mediators of apoptotic signaling revealed up-regulation of BAX expression and caspase-3 activity, as well as significant decreases in Bcl2 and P-Akt. We conclude that MK886 reduces the viability of leukemia HL-60 cells by reducing mPGES-1 expression and PGE2 synthesis in a dose-dependent manner, which strongly suggests that mPGES-1 inhibitors should be considered as promising candidates for leukemia treatment.

Nanocrystalline La2Sn2O7and La2Sn1.8Co0.2O7with a phase-pure pyrochlore structure were synthesized by a hydrothermal method, and their catalytic activity was investigated for soot combustion. The as-synthesized catalysts presented relatively larger surface area, and pore volume, which was benefit to the gas molecule diffusion in the reaction. A uniform spherical structure with particle size of 200–500 nm was found in SEM. The samples via hydrothermal route are more active for catalytic soot combustion, ascribing to the spherical morphology, high surface area and improved oxygen mobility. After Co, the reducibility was improved and surface oxygen vacancy was produced, resulting in the enhanced activity and selectivity to CO2formation.

Our previous studies have shown that quercetin inhibits Cox-2 and Bcl-2 expressions, and induces human leukemia HL-60 cell apoptosis. In order to investigate the role of AMP-activated protein kinase (AMPK) on quercetin-induced apoptosis of HL-60 cells, we used flow cytometry to detect cell apoptosis. The expressions of LKB1, phosphorylated AMPK (p-AMPK), and Cox-2 protein were detected in HL-60 cells and normal peripheral blood mononuclear cells (PBMCs) by western blot. The expressions of LKB1, p-AMPK, and Cox-2 were detected in HL-60 cells after culture with quercetin. The expressions of p-AMPK were detected in HL-60 cells after culture with AMPK inhibitor Compound C. Then, the expressions of LKB1, p-AMPK, and Cox-2 were detected in HL-60 cells after culture with quercetin alone or quercetin + Compound C. It was found that there was no significant difference in LKB1 between PBMCs and HL-60. p-AMPK in PBMCs was higher than that in HL-60, while Cox-2 was lower. After culture of HL-60 with quercetin, p-AMPK was increased, Cox-2 was decreased, but LKB1 remained unchanged. After culture of HL-60 with Compound C, p-AMPK was decreased. There was no significant difference in LKB1 between the quercetin-alone and the quercetin + Compound C groups. p-AMPK decreased more significantly, while Cox-2 increased more significantly in the quercetin + Compound C groups than those in the quercetin-alone groups. Taken together, these findings suggested that quercetin activates AMPK expression in HL-60 cells independent of LKB1 activation, inhibits Cox-2 expression by activating AMPK, and further regulates the Bcl-2-dependent pathways of apoptosis to exert its anti-leukemia effect.

Resistance to chemotherapy is still a challenge for the treatment of acute myeloid leukemia. Combination use of histone deacetylase inhibitors (HDACIs) and proteasome inhibitors may provide a potential way to overcome drug resistance. One of the HDACIs, valproic acid (VPA), and a proteasome inhibitor, bortezomib (BOR), were assessed. Co-exposure of cells to VPA and BOR inhibited proliferation, arrested the cell cycle in G0-G1 phase and induced apoptosis in both HL60 and HL60A cells. These events were accompanied by the inhibition of cyclin D1 and human telomerase reverse transcriptase (hTERT) as well as telomerase activity. Moreover, synergism of proliferation inhibition was found in HL60A, superior to the additivity in HL60. The effects of combination treatment on cell cycle arrest and telomerase activity inhibition in HL60A were also more striking than those in HL60. In summary, our findings provide an insight into future clinical applications of the VPA-BOR combination regimen for AML, especially in those cases which are resistant to conventional chemotherapy.

In this paper, the sorbent bentonite and zeolite was used to treat the wastewater including basic brilliant green dye were investigated respectively. During this experiment, the quantity of sorbent, the surge time, the concentration of basic brilliant green dye and the pH of the solution, which make different effect on the rate of absorption, have been researched. And the remarkable adsorption conditions of the two sorbents were made certain. The conclusions indicated that when the quantity of bentonite was 25 mg and the surge time was 30 minutes, the absorption rate was over 94%, and when the quantity of zeolite was 180 mg and the surge time was 30 minutes, the absorption rate reached over 92%. So it shows that the adsorption effect of the bentonite is superior to the zeolite, and all they have wide application market.

Quercetin is one of the naturally occurring dietary flavonol compounds. It is present abundantly in plants and has chemopreventive and anticancer effects. To investigate its anticancer mechanism, we examined the activity of quercetin against acute leukemia cell line, HL-60. Our results showed that quercetin inhibited cell proliferation and induced apoptosis in a time- and dose-dependent manner. Furthermore, quercetin down-regulated the expression of anti-apoptosis protein Bcl-2 and up-regulated the expression of pro-apoptosis protein Bax. Caspase-3 was also activated by quercetin, which started a caspase-3-depended mitochodrial pathway to induce apoptosis. It was also found that quercetin inhibited the expression of the cycloocygenase-2 (Cox-2) mRNA and Cox-2 protein. Taken together, these findings suggested that quercetin induces apoptosis in a caspase-3-dependent pathway by inhibiting Cox-2 expression and regulates the expression of downstream apoptotic components, including Bcl-2 and Bax. Quercetin can be a potent and promising medicine which might be safely used in leukemia therapy.

Abstract 4824 Recent studies have shown that prostaglandin E2 (PGE2) may play a key role in the tumorigenesis and tumor development. Membrane-bound prostaglandin E2 synthase-1 (mPGES-1), an inducible enzyme that acts downstream of cyclooxygenase (COX) and specifically catalyzes the conversion of prostaglandin H2 (PGH2) to PGE2, was over-expression in a variety of solid tumor cells and tissues such as nonsmall-cell lung cancer, colon carcinoma, gastric carcinoma and breast cancer. MK886, a small molecular inhibitor, is a reasonable potency as an inhibitor of mPGES-1 in vitro experiment. In this study, we examined effects of MK886 on expression of mPGES-1 and PGE2 synthesis in human acute myeloid leukemia cell line (HL-60), observed cell proliferation and apoptosis after 24-h treatment with MK886, and tried to explore the possible mechanisms by checking some protein belong AKT cell singling pathway such as P-AKT, Bax and Bcl-2. We found that the expression levels of mPGES-1 mRNA and protein were higher in HL-60 cells than in normal mononuclearcells (MNC). MK886 inhibited mPGES-1 mRNA and protein expression and reduced PGE2 secretion in HL-60 cells in a dose-dependent manner. The cell proliferation was inhibited and the IC50 was 132.16μmol/L. With the increase of MK886 concentration, the cell apoptosis rate assayed by flow cytometry increased and the apparent apoptotic bodies increased when staining by Hoechst 33258. After treated with MK886 for 24h, protein was extracted and assayed by western blot. The results showed that the expression levels of P-AKT, Bcl-2 and c-myc decreased while the Bax protein expression increased in a dose-dependent manner. The caspase-3 activity, determined by colorimetric detection, also increased dose-dependently. These results indicated that mPGES-1 over-expressed in leukemia cell line HL-60, MK886 could induce apoptosis in HL-60 cells via reducing mPGES-1 expression and PGE2 synthesis dose-dependently, thereby regulate the AKT pathway including Bcl-2 family and the activity of caspase-3. It suggested that mPGES-1 inhibitor might emerge as an important therapeutic tool for leukemia treatment. Disclosures: No relevant conflicts of interest to declare.

Abstract 4749 To compare the cytotoxicity of NK cells on RPMI-8266 multiple myeloma cells before and after thermotherapy, and to explore its mechanism. MethodF RPMI-8226 cell was heated for 2 hours at 42°C and after continous culturing for 48 hours, remove dead cells at lower-speed centrifugation. Detected cytotoxicity of NK cells against RPMI-8226 cells, combinig with or without thermotherapy, by LDH releasing assay at different effect-to-target cell ratios (E:T). Moreover compare the expression of HLA class I molecules and NKG2D ligands-MHC class I chain-related molecule A or B and human cytomegalovirus glycoprotein UL16 binding proteins, ULBPsFULBP1AULBP2AULBP3 on the surface of RPMI 8226 cells were analyzed by flow cytometry. Result As for E:T is 5:1, the cytotoxicity of NK cells combining with or without hyperthermia on the 8266 NK cells are (2.15 ± 0.32)% and (9.32 ± 1.27)% respectively; at E:T(10:1), cytotoxicities are (5.26 ± 0.84 )% and (20.37 ± 1.78)%; at E:T (20:1), they are(7.63 ± 1.05)% and (28.69 ± 2.25)%;at ET(40:1), they are (10. 18 ± 1.53)% and (35.64 ± 4.87)%. At the different target ratio, the cytotoxcity of NK cells combining with or without thermotherapy were significant statistical difference (P 0.05). Conclusion Hyperthermia can enhance cytotoxicity of NK cells on multiple myeloma RPMI-8266 cells through upregulating the expression of NKG2D ligand ULBP1 and ULBP3 molecule on RPMI-8266, while no effect on HLA ± molecules. Disclosures: No relevant conflicts of interest to declare.

Vascular endothelial growth factor (VEGF), which induces angiogenesis and increases vascular permeability, is a major growth factor mediating tumor progression. In this study, we employed immunohistochemical-staining method to detect the expression of VEGF in lymph nodes taken from39 non-Hodgkin’s lymphomas patients and analyzed the relation of the expression levels to malignant aggressiveness, treatment response, histological grade, clinical stage and prognosis. The patients had been observed for at least 5 years or until death. 9 patients with benign lymphadenopathy were acted as control. The expression of VEGF was assessed according to the percentage of immunoreactive cells in a total of 1000 neoplastic cells (quantitative analysis). Immunoreactivity was graded positive, more than 10% of carcinoma cells stained and negative, no detectable staining or less than 10% of carcinoma cells stained. Furthermore, the qualitative intensity of staining for VEGF was assessed using a scale of 0–3+. The expression analysis of VEGF revealed that in 31 out of 39 (79.49%) specimens VEGF staining was positive. The VEGF staining was always cell membrane. Significant associations were found between the expression of VEGF and histological grade, Ann Arbor stage, prognosis (according to International Prognostic Index, IPI) and chemotherapy response. Among 8 cases of low grade, 7 had lower-level expression and 1 had higher-level expression, but among 31 cases of intermediate and high grade, 13 had lower-level expression and18 had higher-level expression (P=0.044

Multidrug resistance (MDR) was the main cause of treatment failure in leukemic chemotherapy. P170, the protein expressed from MDR-1 gene, play an important role in MDR. P27Kip1 was a member of cyclin dependent kinase inhibitor family. It was also a negative regulator in cell generation cycle. It also promoted cells apoptosis. Valproic acid (VPA) was a kind of histone deacetylace inhibitor which could inhibit cell cycle and induce apoptosis. In the current study, we investigated the effects of VPA on the expression of P27Kip1 and P170 of HL60 cells in vitro. HL60/HT cells were induced from HL60 cells by harringtonine (HT) in gradient concentrations. The P27Kip1 expression and P170 expression were tested in flow cytometry. We tested the expression of P27Kip1 and P170 in normal mononuclear cells, HL60 cells and HL60/HT cells, also the effects of VPA on P27Kip1 expression and P170 expression in HL60 cells and HL60/HT cells. The 50% inhibiting concentration (IC50) of vincristine for HL60/HT cells was 5.2 times higher than that for HL60 cells (0.078ug/ml vs. 0.015ug/ml). The IC50 of HT for HL-60/HT cells was 9.3 times higher than that for HL-60 cells (0.14μg/ml vs. 0.015μg/ml). The IC50 of Cytarabine for HL60/HT cells was 3.65 times higher than that for HL60 cells (0.73ug/ml vs. 0.20ug/ml). The expression of P27Kip1 in normal mononuclear cells was the highest, while that in HL60 cells was higher than HL60/HT cells (96.18±3.13% vs. 85.83±4.65% vs. 35.91±3.13%, one way ANOVA, P0.05). But the P170 expression of HL60/HT cells was 6.71±1.61%, which was much higher than that of normal mononuclear cells or HL60 cells (P0.05). In HL60/HT cells, the expression of P27Kip1 increased after cultured with VPA (66.20±5.07% vs. 35.91±3.13%, P0.05). These results indicate that the P27Kip1 expression in normal mononuclear cells were higher than that in leukemia cells (HL60 cells), and the P27Kip1 expression in common HL60 cells were higher than that in MDR HL60/HT cells. VPA could increase the expression of P27Kip1 in HL60 cells and HL60/HT cells. VPA had no effect on the expression of P170 in HL60 cells or HL60/HT cells. That means, The MDR-reverse effect of VPA may not come from the P170 descent mechanism.

Platelet activation, including platelet adhesion, platelet aggregation and platelet release reaction, played an important role in thrombogenesis. We all knew that Platelet glycoprotein IIb/IIIa antagonist was the most effective drug for anti-aggregation, while we don’t know clearly its effect on platelet release reaction and the relations between its effects on platelet aggregation and release reaction. Platelet release reactions included α-granules and dense granules releasing. When α-granules were released, its membrane glycoprotein CD62p was expressed in the platelet membrane. We used the CD62p expression as the index of platelet release reaction. In the current study, the 4-peptides RGDS (Arg-Gly-Asp-Ser) was used as glycoprotein IIb/IIIa antagonist. We detected the effects of RGDS on platelet aggregation and CD62p expression induced by adenosine diphosphate (ADP) (finial concentration, 5μmol/L) in vitro. 50, 100, 200, 400 and 800μmol/L RGDS were used separately in the test. RGDS of each concentration could significantly inhibited maximal platelet aggregation (PAG(M)) induced by ADP, the 50% inhibiting concentration was approximately 200μmol/L. 800μmol/L RGDS could inhibited PAG(M) by 80.48±8.18%. Only ≥200μmol/L RGDS could significantly inhibited platelet CD62p expression. 800μmol/L RGDS could inhibit platelet CD62p expression by 27.31±9.74%. The inhibiting effect of RGDS on PAG(M) and platelet CD62p expression had significantly correlation (r =0.976, P

Background: Platelet activation plays an important role in cardiovascular complications which are frequently seen in patients with diabetes mellitus. It has been demonstrated that platelets are hyperactive in diabetes patients and this is supposed to be due to high platelet cytoplasmic free calcium ion concentration ([Ca2+]i). There are chloride channels on the membranes of platelets and the activation of chloride channels may participate in the transportation of calcium. Our previous study also showed that niflumic acid (NFA), a chloride channel blocker, can inhibit the elevation of platelet [Ca2+]i in healthy volunteers. The goal of this study was to investigate effects of NFA on platelet [Ca2+]i and platelet aggregation rate (PAG) in diabetic patients. Methods: We prospectively enrolled 17 consecutive patients with diabetes mellitus at diagnosis. The control group was formed from 17 healthy volunteers. Platelet [Ca2+]i was detected by Fura-2 fluorescent technique and PAG was detected by platelet aggregometer. We studied the effects of NFA, nifedipine, a calcium channel blocker, and their combining effects on platelet [Ca2+]i and PAG in diabetic patients. Results: The PAG, platelet resting [Ca2+]i, Ca2+ release and Ca2+ influx in diabetic patients were significantly higher than control group(P Conclusions: Platelets in patients with diabetes mellitus are hyperactive. NFA reduces the PAG and platelet [Ca2+]i influx possibly by blocking the chloride channels on platelet membrane in diabetic patients. There are no interactions between NFA and nifedipine on the movement of platelet [Ca2+]i.

Objective To explore the effects and interactions of GPIIb/IIIa antagonists and chloride channel blockers on the platelet cytoplasmic free calcium ([Ca2+]i ). Methods We washed and suspended fresh platelets with Hepes buffer containing 0.1% bovine serum albumin (BSA), then loaded platelets with 5μmol/L Fura-3/AM. Then RGDS, the GPIIb/IIIa antagonists, and the chloride channel blockers 4,4′-diisothiocyano-2,2′-disulfonic acid stilbene(DIDS) or niflumic acid(NFA) were added to the platelet suspension. After 2 minutes incubating, we observed the effects and interactions of GPIIb/IIIa antagonists and chloride channel blockers on platelet [Ca2+]i by measuring the Fura-3 fluorescence intensity. Results 1. Effects of GPIIb/IIIa antagonists and chloride channel blockers on platelet [Ca2+]i. The fluorescence intensity of resting platelet [Ca2+]i were 369.6±62.2, 381.9±72.4, 392.8 ±69.9 after adding RGDS(250 μmol/L), DIDS(100 μmol/L) or NFA( 100 μmol/L) respectively. every agent had no significant effect on resting [Ca2+]i (p>0.05). After thrombin(0.03 U/ml) stimulating and adding RGDS(250 μmol/L), DIDS(100 μmol/L) or NFA( 100 μmol/L), the platelet [Ca2+]i were 883.9±107.0, 789.8±99.8, 564.1±79.4. Compare with the control(977.9±108.8), the three agents could inhibit the elevation of [Ca2+]i stimulated by thrombin (p 2. Combined effects of GPIIb/IIIa antagonists and chloride channel blockers The fluorescence intensity of resting platelet [Ca2+]i was 383.9±67.9 after incubated with RGDS and DIDS. That had no significant effects. When platelets were stimulated by thrombin (0.03 U/ml), the combined inhibition rate was (24.4±10.8)%, RGDS and DIDS couldn’t weaken or enhance each other on thrombin-induced elevation of [Ca2+]i (p>0.05). Neither RGDS nor NFA had significant combined effects on resting [Ca2+]i(p>0.05). The combined inhibition rate was (46.0±7.3)%, they had no interactions too(p>0.05). Conclusion The GPIIb/IIIa antagonists RGDS and the chloride channel blockers DIDS or NFA have no effect on resting platelet [Ca2+]i. All of them can inhibit the elevation of platelet [Ca2+]i induced by thrombin. There are no interactions between GPIIb/IIIa antagonists RGDS and chloride channel blockers (DIDS or NFA) in resting platelet [Ca2+]i and elevation of platelet [Ca2+]i induced by thrombin, and their effects were independent.