Cytotoxic effects of docetaxel as a candidate drug of drug-eluting stent on human umbilical vein endothelial cells and the signaling pathway of cell migration inhibition,adhesion delay and shape change

Docetaxel(DTX),a paclitaxel analogue,can efficiently inhibit proliferation of vascular smooth muscle cells and has broadly been used as an antiangiogenesis drug.However,as a candidate drug of drug-eluting stent,the effects of DTX on human umbilical vein endothelial cells(HUVECs)are still not well understood.Herein,we investigated the effects of DTX on proliferation,apoptosis,adhesion,migration and morphology of HUVECs in vitro.We found that DTX had the cytostatic and cytotoxic effects at low and high concentrations,respectively.DTX could inhibit the proliferation and migration of HUVECs,induce HUVECs apoptosis,delay HUVECs adhesion and decrease spreading area and aspect ratio of individual cells.The signaling pathway that DTX led to the migration inhibition,adhesion delay and shape change of HUVECs is the VE-cadherin mediated integrin b1/FAK/ROCK signaling pathway.The study will provide a theoretical basis for the clinical application of DTX.

SUPEROXIDE RADICAL SCAVENGING AND CYTOTOXIC EFFECTS OF DIFFERENT MARINE SPECIES FROM TURKEY'S COASTS

In this study,acetone extracts of thirteen different species distributed in Turkey Coasts including tunicates,sea anemones,sponges and corals were investigated for their superoxide(SO)radical scavenging and cytotoxic activities.VWhile SO radical scavenging activity was tested using alkaline DMSO method,cytotoxic activity was tested by MTT assay against Hep-2 cancer cell line.As a result of bioactivity studies,Paramuricea clavata extract showed

Dynamic monitoring of the cytotoxic effects of protoberberine alkaloids from Rhizoma Coptidis on HepG2 cells using the xCELLigence system

AIM: To investigate the cytotoxic effects of the six protoberberine alkaloids(PAs) from Rhizoma Coptidis on HepG2 cells. METHOD: A systematic screening was conducted to investigate the dynamic response of HepG2 cells to the PAs using the impedance-based xCELLigence system. Cisplatin was selected as the positive control. The real time, concentration-response curves and the 50% inhibitory concentrations(IC50) were acquired to evaluate the anticancer activity of the PAs. RESULTS: All of the six PAs inhibited cell growth and induce death in HepG2 cells in a time- and concentration-dependent manner. The IC50 values of cisplatin, berberine, columbamine, coptisine, epiberberine, jatrorrhizine, and palmatine were 5.13, 42.33, 226.54, 36.90, 302.72, 383.54, and 456.96 μg·mL-1, respectively. The results obtained using the xCELLigence system corresponded well with those of the conventional methods. CONCLUSION: The xCELLigence system is a reliable and efficient tool for real-time screening of the cytotoxic effect of compounds in cell-based in vitro assays. Coptisine and berberine, with methylenedioxy group at C2 and C3 on the phenyl ring showed stronger effect.than the other four PAs. However, compared with cisplatin, the six PAs didn't show obvious cytotoxic effect on HepG2 cells.These results provided some useful data for the evaluation of the anticancer compounds, and the clinical application of traditional Chinese medicine.

Cytotoxic Effects of Biodegradation of Pure Mg and MAO-Mg on Tumor Cells of MG63 and KB

Magnesium （Mg） based metals possess unique features of biodegradation that can be used for different purposes. In this study, two tumor cells, MG63 and KB, were cultured directly on a pure Mg with and without MAO coating to evaluate their cytotoxic effects on the tumor cells. It was found that both MG63 cells and KB cells adhered better on the surface of the coated Mg samples than those on the pure Mg samples. However, the survivals of two kinds of tumor cells were obviously inhibited on both Mg samples with and without coating, compared with that on the pure titanium. In addition, the survival of MG63 cells was more susceptive to the Mg degradation compared with the KB cells. Thus, the application of such effect should take account of that the degradation of Mg based metal may result in different cytotoxic effect on different cells.

EFFECTS OF PHYTOLACCA ACINOSA POLYSACCHARIDES I ON CYTOTOXICITY OF MACROPHAGES AND ITS PRODUCTION OF TUMOR NECROSIS FACTOR AND INTERLEUKIN 1

The in vivo effects of Phytolacca acinosa poly-saccharides I (PEP-I) on immunologic cytotoxicity of mouse peritoneal macrophages and its production of tumor necrosis factor (TNF) and interleukin 1 (IL-1) were studied. PEP-I 80 or 160 mg kg was given ip twice every 4 day. Both doses were found to have significant enhancing activity on macrophages cytotoxicity against S180 sarcoma cells and malignant transformed fibroblast L929 cells. Peritoneal activated macrophages were incubated with LPS for 2 and 24 hrs to induce TNF and IL-1, respectively. The TNF and IL-1 activities were tested from cytotoxicity against L929 cells in an absorbence assay of enzymatic reaction and proliferation of thymocytes co-stimulated assay separately. The optimal time for TNF production was found on day 8. Significant increases in TNF and IL-1 were observed. In comparison of the effect of PEP-I on TNF with that of known priming agent BCG, there was no difference between them, but PEP-I had a high effect on IL-1. These results suggest that cytotoxicity of macrophages primed by PEP-I is closely related to its TNF and IL-1 production.

Cytotoxicity assessment of a gold nanoparticle-chitosan nanocomposite as an efficient support for cell immobilization:comparison with chitosan hydrogel and chitosan-gelatin

Cell-based biosensors have become a research hotspot in the biosensors and bioelectronics fields. The main feature of cell-based biosensors is immobilization of living cells on the surface of transducers. Different types of polymers which are used as scaffolds for cell growth should be biocompatible and should have reac- tive functional groups for further attachment of biomolecules. In this work, cell attachment and proliferation on chitosan hydrogel, chitosan-gelatin and gold nanoparticle-chitosan nanocomposite membranes was stud- ied. Characterization of the membranes was performed using scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR). Cytotoxicity assessment on HEK293 cells was carried out for all mem- branes using the MTT assay. Cell morphology and viability were assessed to evaluate cell attachment and pro- liferation. Regarding cell studies, the findings revealed that none of the membranes induced cytotoxic effects. However, the data showed that gold nanoparticle nanocomposite membranes improved HEK293 attachment and adhesion more than other membranes, indicating that it provides an effective surface for immobilizing cells for sensing applications.

THE ANTICANCER EFFECT AND ANTI－DNA TOPOISOMERASE II EFFECT OF EXTRACTS OF CAMELLIA PTILOPHYLLA CHANG AND CAMELLIA SINENSIS

The cytotoxic effect of extract of camellia ptilophyllachang(ECPC) and extract of camellia sinensis(ECS) onHeLa cell line, poorly differentiated nasopharyngealcarcinoma cell line(CNE2) and gastric cancer cell line(MGC-803 ) in vitro was studied using MIT assay method.The results showed that ECPC and ECS possessed significantcytotoxic effect on above three cell lines. The anticancer testin mice showed that ECPC had marked inhibitory effectagainst Ehrlich solid carcinoma(ESC) with inhibition ratesof 17. 8 48. 3% and with inhibition rates of 28. 3-54. 5% against reticular cell sarcoma(L2), and that ECShad inhibition rates of 31 . 5 -49. 4 % against ESC and 35. 8- 50% against L2. These two extracts had only marginalinhibitory effect against sarcoma- 180. The unknottingactivity of DNA topoisomerase II was inhibited completelyby ECPC and ECS at the concentration of 50 μg/ mlsuggesting that DNA topoisomerase II might be a targetenzyme of these two extracts.

Mechanisms of rotenone-induced neurotoxicity in PC 12 cells

BACKGROUND： Rotenone-induced neurotoxicity in PC 12 cells has been widely used to study the pathogenesis of Parkinson＇s disease. However, the precise mechanisms underlying rotenone-induced dopaminergic neuronal degeneration in Parkinson＇s disease remains unclear. OBJECTIVE： To establish rotenone-induced neurotoxicity in PC 12 cells, and to investigate the possible action pathways to rotenone-induced neural cell injury at the protein level. DESIGN, TIME AND SETTING： A controlled proteomics study was performed at the Department of Neurology, First Hospital, Jilin University between March 2006 and March 2007. MATERIALS： PC 12 cells were obtained from Shanghai Cell Bank of Chinese Academy of Sciences, China. Rotenone was provided by Sigma, USA. METHODS： PC 12 cells in logarithmic growth phase were treated under experimental and control conditions, respectively. A total of 0.5 μmol/L rotenone, or the same amount of Dulbecco＇s modified eagle＇s medium （DMEM）, was added in the experimental and control conditions, respectively. MAIN OUTCOME MEASURES： Following 72 hours of rotenone treatment, cellular survival rate was determined by methyl thiazolyl tetrazolium assay, and apoptotic changes were detected by Hoechst 33342 staining. Total cellular protein was extracted to acquire differential protein expression data utilizing two-dimensional differential in-gel electrophoresis. To identify differential protein spots, matrix-assisted laser desorption/ionization-time of flight mass spectrometry （MALDI-TOF-MS） was used. RESULTS： In the MTT assay, the experimental condition induced significantly less cell survival compared to the control condition （P 〈 0.01）. Hoechst 33342 staining revealed a larger number of apoptotic cells under the experimental condition compared to the control condition （P 〈 0.01）, as determined by the presence of nuclear condensation, pyknosis, and nuclear fragmentation. Two-dimensional electrophoresis results showed that the differential expression of protein spots 1069 and 1538 was increased by 144% and 124%, respectively, while that of protein spot 1094 was decreased by 123% in the experimental condition compared to the control condition （P 〈 0.01）. By MALDI-TOF-MS analysis and database retrieval, γ-enolase, triosephosphate isomerase 1, and eukaryotic translation initiation factor 4A were confirmed to be involved in rotenone-induced neural cell injury. CONCLUSION： γ-enolase, triosephosphate isomerase 1, and eukaryotic translation initiation factor 4A might participate in rotenone-induced neurotoxicity in PC 12 cells.

Octacyclic and decacyclic ent‑abietane dimers with cytotoxic activity from Euphorbia fischeriana steud.

A novel Diels-Alder adduct possesses a 6/6/6/5/6/6/6/6 octacyclic skeleton featured with bicyclo[2.2.2]octane moiety,biseupyiheoid A(1),along with another decacyclic 6/6/6/3/5/6/5/6/6/6 fused diterpenoid dimer,bisfischoid C(2),were isolated from Euphorbia fischeriana.Their structures were determined by spectroscopic,X-ray crystallographic approaches,and quantum mechanical calculations.The structural features of 1 and 2 were hypothesized to involve intramolecular Diels-Alder reactions with different coupling patterns.Dimer 1 showed antiproliferative activity through apoptosis activation in LoVo cells.

Synergistic Cytotoxicity of Ampelopsin Sodium and Carboplatin in Human Non-Small Cell Lung Cancer Cell Line SPC-A1 by G1 Cell Cycle Arrested

Objective： To evaluate the cytotoxic effects of ampelopsin sodium（Amp-Na） and carboplatin（CBP） used alone or in combination on human non-small cell lung cancer（NSCLC） cells SPC-A1 in vitro and its related mechanism. Methods： Cytotoxic effects were assessed by 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide（MTT） assays. The synergistic effects of the drugs were calculated with coefficient of drug interaction（CDI）. Cell cycle was determined by flow cytometry（FCM）. The levels of p53, p21, cyclin E, cyclin D1, and phosphorylated cyclin-dependent kinase-2（p-CDK2） were evaluated by Western blot. Results： Amp-Na（6.25–200 μg/m L） and CBP（3.13–100 μg/m L） alone exhibited prominent cytotoxic activity in a concentration-dependent manner on SPC-A1 cells with 50% inhibitive concentration values of 57.07±14.46 and 34.97±6.30 μg/m L, respectively. Drug combinations were associated with significantly higher cytotoxic effects than each drug alone（P〈0.05 or 0.01）. The CDI analysis confirmed the synergy of Amp-Na and CBP on inhibiting cancer cell viability across a wide concentration range（CDI 〈1）. FCM and Western blot showed that synergistic cytotoxic effects of Amp-Na and CBP were related to G1 arrested which mainly mediated by p21 through the inhibition of CDK2 activity independent of the p53 tumor suppressor pathway. Conclusions： AmpNa exhibits anticancer activities and enhances the antitumor activities of CBP through up-regulation of p21 and inhibition of CDK2 activity in human NSCLC cells SPC-A1. These results suggest that Amp-Na may be applied to enhance the anticancer action of CBP.

Acid dissociation constants and cytotoxicity test of a series of omega-aminoalkyl phosphates

We synthesised a series of ω-aminoalkyl sodium hydrogen phosphates （AAP-n-Na, n =3, 4, 5, 6, purity 〉 99%）, which have potential applications as bioactive cosmetic ingredients and surface modifiers of bone minerals （i.e. hydroxyapatites）. Results from Fourier transformed infrared （FTIR）, nuclear magnetic resonance （NMR） and high resolution mass spectroscopy, and elemental analysis all matched their chemical structures. The acid dissociation constants （pKa＇s） of each AAP-n （acid form of AAP-n-Na, n ; 2-6） were measured by potentiometric titration, showing a general increasing trend with an increase in the chain length of AAP-n. However, the pKa3 constant, which corresponds to the deprotonation of the ammonium group in AAP-n-Na, displayed an unusual decrease when n = even. This odd-even effect can be explained by the pairwise self-association of AAP-n-Na molecules in water where intermolecular hydrogen bonding in case of n=even is weaker than that in case of n=odd. All AAP-n-Na at concentrations up to 0.1% （w/v） were non-toxic to L929 fibroblasts and MG 63 osteoblast-like cells in terms of cell growth and morphology, These basic data were important for applications of AAP-n and their salts in biomedical engineering.

An analytical method for quantitative estimation of the cytotoxicity of dental alloys using MTT assay,ICP analysis and effective cytotoxicity calculation

The two most important criteria for dental materials are their biofunctional and biocompatible endurance within the anticipated life-span of the dental restoration in the mouth. Biocompatibility relates mainly to the allergenicity and the toxicity of the material. To test the non-specific toxicity of dental materials, in vitro cell culture assays have been developed. For in vitro screening, such tests are recommended to check the cytotoxicity of dental materials (ISO 10993 5). Various studies have already been performed to quantitatively determine the cytotoxicity level of dental alloys. However, as long as only dental alloys and the cell culture technique are applied, it is not possible to determine which of the alloying elements cause the cytotoxicity. Therefore, an analytical method is needed. Wataha et al determined in 1991 the TC50 values of 9 metal cations of various dental casting alloys, using cell culture methods. Kapert et al reported in 1994 a complex in vitro test concept, where the ICP analysis (inductively coupled plasma emission spectroscopy) was introduced to measure the trace elements extracted from various alloys. Experimentelle Zahnheilkunde, Universitts ZMK Klinik Freiburg, Germany (Lü XY and Kappert HF) The aim of the present study was to find a relation between the ICP results, the TC50 value of metal cations, and the cytotoxicity of dental alloys. The cytotoxicity levels of various dental alloys and the TC50 values of 10 metal cations were established using the MTT assay, an effective cell culture of method. Then, the concentrations of the corrosively soluted metal cations in the extracts of the alloys were measured using the ICP method. From all these experimental results it was found that the relation between the effective cytotoxicity Z eff of an alloy, the concentrations C i of i th trace element and the TC50 values T Ci of the i th metal cation can approximately be expressed by Z eff =∑iC i2·T Ci . Two significant applications of this expression are a) The cytotoxicity of an alloy can be estimated by ICP analysis of the extract if the TC50 values of the trace elements are know. b) The cytotoxicity of a new-developed-alloy can be estimated in advance, according to the alloying components.

Parasporal Proteins from Bacillus thuringiensis and Their Cytotoxicity on Human Cancer Cells

Parasporins(PSs) represent a novel functional category of crystal proteins(Cry) produced by non-insecticidal Bacillus thuringiensis. A distinct feature for PSs is their specific cytotoxicity against human cancer cells from diverse origins, other than hemolytic or insecticidal activity. As structurally/functionally Cry proteins, parasporins are expressed as protoxins that require protease cleavage for activation. Currently,identified PSs is classified into 6 groups: PS1, PS2, PS3, PS4, PS5 and PS6, which are heterogeneous in cytotoxic spectrum and activity level. Some PSs have been explored for their mode of anticancer activities, reports mainly include pore formation induced by binding to putative receptors on cell membrane and apoptosis by intracellular Ca2+concentration. Further work should focus on the identification of new PS or PS homologs and better understanding of their anticancer mechanism before possible application in cancer therapy.

Immune mechanism and clinical significance of macrophage to medullary hematopoietic injury of immune-related hematocytopenia patients

Background Immune-related hematocytopenia （IRH） is considered to be related with the production of autoantibody, as well as the activation of humoral immunity which is stimulated by B lymphocyte. This study aimed to observe the levels of various cytokines in the blood serum and the in situ active state of macrophage （Me） in the medullary hematopoietic microenvironment of IRH patients, and to probe into the immune mechanism and clinical significance of Me in hematopoietic cell injury. Methods ELISA is used to detect the IL-4, IL-6, IL-12, IL-17, and IFN-y levels in the peripheral blood serum of 376 patients in pre- and post-therapy. Cytochemistry and cell immunochemistry methods are used to observe the peroxidase （POX）, nonspecific esterase （NSE）, hemosiderin granules, and HLA-DR activity of Me in the bone marrow of patients. Immunofluorescence is used to observe the expression of hemocyte antihuman globulin IgG antibody, lymphocytes CD4 molecule, Me membrane Fcyllreceptor （FcyllR）, mannitose receptor （MR）, IFN-y, ICAM-1, IL-12, and IL-17A and the formation mechanism of antibody-dependent cell-mediated cytotoxicity （ADCC） hematopoietic cell islands （HI） in the medullary hematopoietic microenvironment of patients. Glucocorticoid is used for treatment on the basis of anti-infection therapy, and gamma globulin stoss therapy is used for the appearance of ADCC-type HI or serious Me bloodthirsty phenomenon; if necessary, association of Cyclosporine A （CsA） should be used and chalybeate should be supplemented. Results In the patient group, the levels of IL-4, IL-6, IL-12, IL-17, and IFN-y were increased. After treatment, the cytokine levels gradually became normal. The activated Me in the marrow highly expressed NSE and POX, and Me swallowed more hemosiderin particles, but the iron in the cytoplasm of immature erythrocytes decreased. The activated Me expressed HLA-DR, MR, ICAM-1, IFN-y, and IL-12. For patients with humoral immunity activation and bacterial infection, Me weakly expressed IL-17A but highly expressed FcyIIR, and the phenomenon that ADCC-type HI broke pathological blood corpuscles often occurred; for the cellular immune activation along with virus infection, the white blood count （WBC） significantly reduced, Me weakly expressed FcyIIR, secretory highly expressed IL-17A, and the phenomena that Me adhered to, captured and swallowed blood cell often occurred. After four weeks of anti-infective and immunosuppressive therapy, nuclear apoptosis of Me occurred in the bone marrow of patients, HI and bloodthirsty phenomenon disappeared, and the peripheral blood picture started to improve. Conclusions Me is an important antigen presenting cell in the IRH marrow for hematopoiesis destruction and an immune effector cell of hematopoietic injury; infection can promote the activation of Me, upregulate the impression of immune molecule and receptors, form ADCC HI. aeGravate hematoBoietic iniurv, and accelerate the destruction on hematoDoietic cell.