Adv

Drug Deliver Rev 2009,61(12):1007–1019.CrossRef 20. Orlova Y, Magome N, Liu L, Chen Y, Agladze K: Electrospun nanofibers as a tool for architecture control in engineered cardiac tissue. Biomaterials 2011, 32:5615–5624.CrossRef 21. Lee KH, Shin SJ, Kim CB, Kim JK, Cho YW, Chung BG, Lee SH: Microfluidic synthesis of pure chitosan microfibers for bioartificial liver chip. Lab Chip 2010, 10:1328–1334.CrossRef 22. Lee HJ, Kim HS, Kim HO, Koh WG: Micropatterns of double-layered nanofiber scaffolds with dual functions of cell patterning and metabolite detections. Lab Chip 2011, 11:2849–2857.CrossRef 23. Tian F, Prina-Mello A, Estrada G, Beyerle A, Moller W, Schulz H, Kreyling W, Stoeger T: Macrophage cellular adaptation, localization and imaging of Screening Library solubility dmso different size polystyrene particles. Nano Biomed Eng 2009, 1:19–38.CrossRef 24. Li Y, Li Z, Zhou BGB324 molecular weight X, Yang P: Detection of nano Eu2O3 in cells and study of its biological effects. Nano Biomed Eng 2010, 2:24–30.CrossRef

25. Lea JY, Song KT, Kim SY, Kim YC, Kim DY, Kim CY: Synthesis and characterization of solublepolypyrrole. Synth Met 1997, 84:137–140.CrossRef 26. ImageJ[http://​rsb.​info.​nih.​gov/​ij] [] 27. Fuh YK, Hsu HS: Fabrication of monolithic polymer nanofluidic channels via near-field electrospun nanofibers as sacrificial templates. Int J Nonlinear Sci Num Simul 2010, 11:979–984.CrossRef 28. Yiin-Kuen F, Li-Chih L, Jason SC: Self-organization of multiple jets in near-field electrospinning process. Jang Micro Nano Lett 2012,7(4):376–379.CrossRef 29. Bhattarai N, Edmondson D, Veiseh O, Matsen FA: Electrospun chitosan-based nanofibers and Rho their cellular compatibility. Biomaterials 2005, 26:6176–6184.CrossRef 30. Wang F, Gao F, Lan M, Yuan H, Huang Y, Liu J: Oxidative stress contributes to silica nanoparticle-induced cytotoxicity in human embryonic kidney cells. Toxicol Vitro 2009, 23:808–815.CrossRef 31. Ji LL, Chen Y, Wang ZT: The toxic

effect of pyrrolizidine alkaloid clivorine on the human embryonic kidney 293 cells and its primary mechanism. Toxicol Pathol 2008, 60:87–93.CrossRef 32. Du J, Che PL, Wang ZY, Aich U, Yarema KJ: Designing a binding interface for control of cancer cell adhesion via 3D topography and metabolic oligosaccharide engineering. Biomaterials 2011, 32:5427–5437.CrossRef 33. Ku SH, Lee SH, Park CB: Synergic effects of nanofiber alignment and electroactivity on myoblast differentiation. Biomaterials 2012. In press 34. click here Arnold M, Adam EAC, Glass R, Blummel J, Eck W, Kantlehner M, Kessler H, Spatz JP: Activation of integrin function by nanopatterned adhesive interfaces. Chem Phys Chem 2004, 5:383–388.CrossRef 35. Maheshwari G, Brown G, Lauffenburger DA, Wells A, Griffith LG, Cell J: Cell adhesion and motility depend on nanoscale RGD clustering. J Cell Sci 2000, 113:1677–1686. 36. Hsu KC, Fuh YK: A novel in-situ roughness measurement based on spatial average analysis of binary speckle image.

0), and the DNA was precipitated with 2.5 M ammonium acetate in ethanol. After two washes with 80% (v/v) ethanol, the DNA pellet was dried and resuspended in 10 μl, 0.2 μl filtrated, double-distilled water. Following the manufacturer’s descriptions the cloning was done by using a Zero blunt TOPO cloning kit (Invitrogen Corporation). Fifty to hundred colonies from each cloning were

picked and sequenced Selleckchem NSC23766 by pyrosequencing. A PYROMark Q96 ID was used to short DNA sequencing of the approximately 40-60 bp clone insert using the recommended protocol (Biotage AB, Uppsala, Sweden) as described previously using the primer PyroBact64f [19]. The sequences (tags) were imported into the software BioNumerics 4.61 and manually checked, aligned and filtered for high quality sequences. Sanger sequencing with an Applied Biosystem PND-1186 chemical structure 3130 Genetic Analyzer (Foster City, CA, USA) was used to check consensus tags for the pyrosequencing accuracy. The Sequence match analysis tool in the Ribosomal database project 10 http://​rdp.​cme.​msu.​edu/​ was used to assign the Phylogenetic position of each consensus tag. The search criteria were for both type and non-type strains, both environmental (uncultured) sequences and isolates, near-full-length

sequences (>1200 bases) of good quality. If there was a consensus at the genus level the tag was assigned this taxonomic classification. If no such consensus was found, the classification proceeded up one level to family and again if no taxonomic affiliation could be assigned the tag continued to be proceeded up the tree as described by Huse et al., [36]. In some cases it was not possible to assign a domain and these sequences might represent new novel organisms or the sequences might be biased, Ribonucleotide reductase in these cases the tags were excluded from the dataset. In total 364 sequences were finally included in the alignment. The

phylogenetic analysis was done by downloading 16S rRNA gene sequences longer than 1,200 base pair from the RDP database of the Ralstonia type strains http://​rdp.​cme.​msu.​edu. The RDP alignment was used and a phylogenetic tree was constructed by using the Ward algorithm in the software Bionumerics. Burkholderia cepacia (GenBank accession no. AF097530) was used as an out-group. Statistics The statistical analysis was done in two steps: First, the association between one predictor at a time and the NEC score was analysed by robust least squares methodology Napabucasin manufacturer adjusting for gestational age. This is equivalent to a normal linear GEE modal with working independence correlation structure on child level. For each predictor the estimated change in expected NEC score is reported with Wald 95% confidence limits in parentheses. The overall association between the predictor and the NEC score is evaluated by a robust score-test. Second, we formulate a normal linear GEE model including gestational age and all predictors with a robust score-test p-value below 0.1 in the above analyse.

Electron micrographs were acquired from uncoated frozen samples, or after sputter-coating with

check details gold three times during 30 s. Micrographs of uncoated samples were taken at an acceleration voltage of 2.5 kV, and consisted of 30 averaged fast scans (SCAN 2 mode). Coated samples were observed at 5 kV using F4 scans. Extraction of nucleic acids DNA was extracted as previously described [28]. RNA from dormant conidia and conidia in early stages of germination (0 and 3 h) was extracted MAPK inhibitor according to Leeuwen and co-workers [29]. RNA from germinating spores (6 and 12 h), mycelia and sporulating mycelia (plate) were extracted according to Plumridge and co-workers [30]. As a final step in both protocols,

the RNA products were purified using a Qiagen RNeasy Mini kit (RNA clean up protocol). Two-hybrid assay The two-hybrid assay was performed using the BACTH System Kit (Euromedex). Full-length cDNA for all six genes were amplified using primers with Selleckchem Mocetinostat internal restriction sites (Table 2). After digestion of the PCR products, the inserts were ligated into linearized and dephosphorylated pKT25 and pUT18C

vectors and used to transform E. coli. All ligations in this work were performed with the ReadyToGo ligation kit (GE Healthcare) and were transformed into NEB 10-β Competent E. coli cells (New England Biolabs), unless otherwise stated. Correct insertions Farnesyltransferase were confirmed with vector specific primers (Table 2) followed by sequencing. Successful clones were co-transformed into electrocompetent BTH101 cells and selected on LA plates supplemented with ampicillin (100 μg/ml) and kanamycin (50 μg/ml). The protein-protein interactions were assayed according to the manufacturer’s protocol with the following modifications. One fresh colony of each interaction was transferred to 100 ml conical flasks with 5 ml LB supplemented with ampicillin 50 μg/ml, kanamycin 50 μg/ml and 0.5 mM IPTG, and incubated with shaking at 100 rpm at 20°C for 72 h. The extent of protein-protein interaction was measured with β-galactosidase assays as units/mg dry weight.

References 1. Moran GP, Sullivan DJ, Coleman DC: Emergence of non Candida albicans Candida species as pathogens. In Candida and Candidiasis. click here Edited by: Calderone RA. Washington DC: ASM Press; 2002:341–348.

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P, Pais C: Epidemiology of candidemia in oncology patients: a 6-year survey in a Portuguese central hospital. Med Mycol 2010, 48:346–54.PubMedCrossRef 7. Saiman L, Ludington E, Pfaller M, Rangel-Frausto S, Wiblin RT, Dawson J, Blumberg HM, Patterson JE, Rinaldi M, Edwards JE, Wenzel RP, Jarvis W: Risk factors for candidemia in

neonatal intensive care unit patients. The National Epidemiology of Mycosis Survey Study Group. Pediatr Infect Dis J www.selleck.co.jp/products/AG-014699.html 2000, 19:319–24.PubMedCrossRef 8. Karlowicz MG, Rowen JL, Barnes-Eley ML, Burke BL, Lawson ML, Bendel CM, Shattuck KE, Horgan M, Albritton WL: The role of birth weight and gestational age in distinguishing extremely low birth weight infants at high risk of developing candidemia from infants at low risk: a multicenter study. Pediatr Res 2002, 51:301A. 9. Clerihew L, Lamagni TL, Brocklehurst P, McGuire W: Candida parapsilosis infection in very low birthweight infants. Arch Dis Child Fetal Neonatal Ed 2007, 92:F127-F129.PubMedCrossRef 10. Muňoz P, Burillo A, Pouza E: Environmental surveillance and other control measures in the prevention of nosocomial fungal infections. Clin Microbiol Infect 2001, 7:38–45.PubMedCrossRef 11. Sautour M, Dalle F, Olivieri C, L’ollivier C, Enderlin E, Salome E, Chovelon I, Vagner O, Sixt N, Fricker-Pap V, Aho S, Fontaneau O, Cachia C, Bonnin A: A prospective survey of air and surface fungal contamination in a medical mycology laboratory at a tertiary care university hospital. Am J Infect Control 2009, 37:189–194.PubMedCrossRef 12.

J Clin Oncol 2006, 24:3871–3879.PubMedCrossRef 20. Zustovich F, Cartei G, Ceravolo R, et al.: A phase I study of cisplatin, temozolomide and thalidomide in patients with malignant brain tumors. Anticancer Res 2007, 27:1019–1024.PubMed 21. Zustovich F, Lombardi G, Della Puppa A, et al.: A phase II study of cisplatin and temozolomide

in heavily pre-treated patients with temozolomide-refractory high-grade malignant glioma. Anticancer Res 2009, 29:4275–4279.PubMed 22. Brandes AA, Basso U, Reni M, et al.: First-line chemotherapy with cisplatin plus fractionated temozolomide in recurrent glioblastoma multiforme: a phase II study of the Gruppo Italiano Cooperativo di Neuro-Oncologia. J Clin Oncol 2004, 22:1598–1604.PubMedCrossRef 23. Kollmannsberger C, Nichols C, Bokemeyer C: Recent advances in management of patients with selleck kinase inhibitor platinum-refractory testicular germ cell tumors. Cancer 2006, 106:1217–1226.PubMedCrossRef 24. Borst P, Rottenberg S, Jonkers

J: How do real tumors become Selonsertib molecular weight resistant to cisplatin? Cell Cycle 2008, 7:1353–1359.PubMedCrossRef 25. Mayer F, Honecker F, Looijenga LH, et al.: Towards an understanding of the biological basis of response to cisplatin-based chemotherapy in germ-cell tumors. Ann Oncol 2003, 14:825–832.PubMedCrossRef 26. Wu S, Chen L, Becker A, et al.: Casein kinase 1alpha regulates an MDMX intramolecular interaction to stimulate p53 binding. Mol Cell Biol 2012, 32:4821–4832.PubMedCrossRef 27. Wu SF, Huang Y, Hou Mephenoxalone JK, et al.: The downregulation of onzin expression by PKCepsilon-ERK2 signaling and its potential role in AML cell differentiation. Leukemia 2010, 24:544–551.PubMedCrossRef 28. Song LP, Zhang J, Wu SF, et al.: Hypoxia-inducible factor-1alpha-induced differentiation of myeloid leukemic cells is its transcriptional activity independent. Oncogene 2008, 27:519–527.PubMedCrossRef

29. Lippert TH, Ruoff HJ, Volm M: Intrinsic and acquired drug resistance in malignant tumors. The main reason for therapeutic failure. Arzneimittelforschung 2008, 58:261–264.PubMed 30. Goldie JH: Drug resistance in cancer: a perspective. Cancer Metastasis Rev 2001, 20:63–68.PubMedCrossRef 31. Shi L, Chen J, Yang J, et al.: MiR-21 protected human glioblastoma U87MG cells from chemotherapeutic drug temozolomide induced apoptosis by decreasing Bax/Bcl-2 ratio and caspase-3 activity. Brain Res 2010, 1352:255–264.PubMedCrossRef 32. Li Y, Li W, Yang Y, et al.: MicroRNA-21 targets LRRFIP1 and contributes to VM-26 resistance in glioblastoma multiforme. Brain Res 2009, 1286:13–18.PubMedCrossRef 33. Ujifuku K, Mitsutake N, Takakura S, et al.: miR-195, PHA-848125 miR-455–3p and miR-10a( *) are implicated in acquired temozolomide resistance in glioblastoma multiforme cells. Cancer Lett 2010, 296:241–248.PubMedCrossRef 34. Bhutia YD, Hung SW, Krentz M, et al.

The Hawksworth (1991) estimate was established based on a ratio of plant:fungal species in temperate regions. Whether these ratios hold up in tropical buy BIBW2992 regions is indirectly assessed in the papers of Berndt (2012) and Mangelsdorff et al. (2012) with sometimes conflicting results, highlighting the value of both sound taxonomic and monographic treatments as well as the need for more long-term fungal check details studies in tropical regions. For instance, the rust fungi (obligate plant pathogens) may be the best documented group of microfungi, yet Berndt (2012) found that ratios of known rust:plant

species in Neotropical countries ranged from 1:16 to 1:124—no doubt a reflection, at least in part, of under sampling for fungi in most of these areas. Lücking (2012) asks the question of not just how many species remain to be discovered, but of what form these species may take. He uses a novel ‘character correlation index’ whereby combinations of traits that are known to be correlated in currently described species are used to predict the traits that Idasanutlin chemical structure are expected to be correlated and found in currently unknown species. He predicts that another 48 lichen-forming fungi in the Graphis group alone remain to be discovered, approximately doubling the

known number in this genus. The impacts of disturbances on fungal communities have been poorly studied in tropical regions, perhaps because these communities have been considered, likely wrongly, as both resistant and resilient to disturbance (Allison and Martiny 2008). Three papers in this issue address Cepharanthine this assumption: da Silva et al. (2012) determine the impact of mining and restoration in Brazilian restinga on communities of arbuscular mycorrhizal fungi by counting and identifying spores. Hattori et al. (2012) show how diversity of polypore fungi is

dependent upon the presence of suitable host trees that may be removed by logging or conversion to plantations in their Malaysian study sites. And, as already discussed, López-Quintero et al. (2012) examine the effects of clearance for shifting cultivation and subsequent forest recovery on fungal diversity. Just as the study of Berndt (2012) shows that species data is unevenly distributed geographically, other papers in this issue show that there are, likewise, a number of specialized habitats that still remain to be fully assessed for tropical fungal diversity. These include fungi inhabiting insect guts, among which are Trichomycetes that have been reviewed by Lichtwardt (2012). The abundance and diversity of insect host species will clearly affect fungal species diversity and an improved assessment of insect-associated fungal diversity in the tropics is certainly a priority for mycologists. Finally, Jones and Pang (2012) provide a timely review of tropical aquatic fungi, highlighting areas in need of future research.

A slight decrease in the degradation rate of R6G occurred with the increase in the recycle number. We observed that the color of the LFP-H microcrystals slightly changed from light gray to dark gray, indicating that oxidation of LFP-H occurred, possibly Fe(II) in LFP-H was transformed to Fe(III) [28]. The slow oxidation of LFP-H during oxidation of R6G might be the reason of the slight decrease in the catalytic activity. In addition, we observed Rabusertib cost that almost no color was changed when LFP-H was stored in an oven at 60°C for one week, indicating that LFH-H is very stable against air oxidation. This high stability of LFP-H in ambient atmosphere is a good advantage for practical

application. Figure BAY 11-7082 ic50 6 Catalytic behavior of the recycled LFP-H particles. Conclusions We report that LFP, which is widely used as an electrode material of a lithium ion battery, can act as an excellent heterogeneous Fenton-like catalyst. The LFP microparticles exhibited much better catalytic activities to decompose R6G than a popular Fenton-like catalyst of

magnetite nanoparticles. The LFP microparticles also showed a good recycling behavior as a Fenton-like catalyst. In addition, the catalytic activities of LFP can be improved by increasing the specific surface area, suggesting that the catalytic activity of LFP can be further improved if nanostructured LFP particles can be properly synthesized. We believe that LFP can be practically used as a catalyst due to its high catalytic activity

and a good recycling behavior. Furthermore, LFP may open new application fields if the catalytic property of LFP is combined with the conventional properties that are useful PTK6 as an electrode of a battery. Acknowledgements This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MEST) (2013M2A8A1041415). Electronic supplementary material Additional file 1: Figure S1: FESEM images. (a) FESEM images of LFP synthesized by hydrothermal method with a slow heating rate of approximately 4°C/min. (b) Magnified FESEM image of (a). Figure S2. Compare of LFP-H and LFP-C in catalytic degradation of R6G. Conditions: 3 g/L of catalyst, 6 mL/L of H2O2 (30%), pH=7. Figure S3. N2 adsorption/desorption isotherms of LFP-C and LFP-H. (DOC 1 MB) References 1. Wang JL, Xu LJ: Advanced oxidation processes for wastewater treatment: formation of hydroxyl radical and application. Crit Rev Environ Sci Tech 2012, 42:251–325.CrossRef 2. Li Y, Sasaki T, Shimizu Y, Koshizaki N: Hexagonal-close-packed, hierarchical amorphous TiO2 nanocolumn arrays: transferability, enhanced photocatalytic activity, and superamphiphilicity QNZ without UV irradiation. J Am Chem Soc 2008, 130:14755–14762.CrossRef 3. Li Y, Sasaki T, Shimizu Y, Koshizaki N: A hierarchically ordered TiO2 hemispherical particle array with hexagonal-non-close-packed tops: synthesis and stable superhydrophilicity without UV irradiation.

However, the effect of RECK silencing in several cancer cells in a hypoxic microenvironment has not been fully identified. Here we investigated that hypoxia suppresses RECK expression

and restoration of RECK by using the strategy of HDAC inhibition inhibits cancer cell migration and invasion. HDAC inhibitors including trichostatin A (TSA) completely restored RECK expression suppressed by hypoxia in the H-Ras MCF10A cell line (human breast cancer) and the HT1080 cell lines (human fibrosarcoma). TSA suppressed the selleckchem activity of MMP-2 and MMP-9 induced by hypoxia and significantly inhibited the hypoxia-stimulated migration and invasion of both cancer cells. RECK overexpression significantly learn more inhibited the hypoxia-induced migration and invasion, suggesting the inhibitory role for RECK in hypoxic conditions. We also demonstrate that silencing of HDAC1 using small interfering (si) RNA suppressed hypoxia-induced RECK downregulation. In conclusion, the inhibition of HDAC successfully restored the expression of RECK under hypoxic conditions. This resulted in the inhibition of cancer cell migration and invasion through the repression of MMP-2 and MMP-9 activity. Poster No. 131 Probing the Role of E-cadherin in

4SC-202 clinical trial Metastasis Using Real-Time Protein Modulation and Intravital Imaging Hon Leong 1 , Shruti Nambiar1, Balaji Iyengar1, BCKDHA Ann F. Chambers1, John Lewis1 1 Oncology, London Regional Cancer Program, London Health Sciences Centre, London, ON, Canada The ability of tumor cells to migrate, invade and intravasate requires the deregulation of interactions with adjacent cells and the extracellular matrix. A major challenge of cancer biology is to observe the dynamics of the proteins involved in this process in their functional and physiologic context. To address this, we developed an E-cadherin chimera fused to both GFP and a FKBP-destabilization domain (DD) that constitutively targets the protein for proteasome degradation until stabilized by SHIELD-1, a small

molecule that binds reversibly to the DD. This approach allows one to dynamically modulate E-cadherin activity at the post-translational level by varying the levels of SHIELD-1. Using the highly metastatic MDA-MB-231-LN cell line, we demonstrate that in the absence of SHIELD-1, E-cadherin is observed only in punctate cytoplasmic vacuole pools that co-localize with 20S proteasome. Within 30 minutes of SHIELD-1 treatment, a shift in localization to the plasma membrane is seen with concurrent formation of cell-cell adherens junctions. SHIELD-mediated induction of E-cadherin significantly reduces cell migration and invasion compared to un-induced MDA-MB-231LN cells expressing the E-cadherin chimera and vector control MDA-MB-231LN cell line.

In addition, the distribution of the gene duplications (Figure 4) revealed that clusters of gene duplications of the same COG function exist on both CI and CII and that most of the gene duplications in a cluster possessed roughly similar levels of sequence conservation. As such, it may be MGCD0103 chemical structure possible that these highlighted chromosomal segments are locally selected for, especially as these gene duplications possess similar functions.

The sequence similarity and evolutionary constraints of the duplicate gene-pair are indicative of the essential or nonessential nature of gene function. Previous studies have revealed shown that the type II topoisomerases gyrase and topoisomerase P005091 IV demonstrated 40 to 60% amino acid sequence identity, but each protein has a distinct function essential for cell survival [55, Selleck Batimastat 56] highlighting the limitations in bioinformatics approaches. In a similar note, duplicate protein pairs with very little amino acid identity can share similar functions. In Bacillus subtilis, the peptide defomylases (Def and YkrB) show similarity only across short sequences (motifs) but both independently carry a deformylase reaction

essential for cell viability [57]. Therefore, gene disruption analysis is further required to determine the definitive function of isologous gene-pairs. In the specific analysis involving the carbon metabolism genes, it is likely that the cluster in CI containing cbbA, cbbF, cbbM, cbbP duplicated first and then cbbG and cbbT duplications arose from CI and were inserted between the duplicated cbbA and cbbP genes on CII. In addition, the two genes that

code for hypothetical proteins found between cbbT and cbbG on CI may have arisen through an additional Astemizole insertion or transposition event. Although these duplicated genes exhibit varying levels of protein divergence, these protein-pairs are under negative selection as evidenced by the functional constraints analysis in Figure 10. Additionally, the identity between the cbbM genes was low (31%). This is most probably due to the high degree of difference between cbbM I and cbbM II . More specifically, it has been shown that cbbM, which performs the first critical step in carbon fixation, has two forms (cbbM I and cbbM II ). The form I enzymes possesses large and small subunits while the form II enzyme possesses only large subunits that are different from the form I large subunits [58]. The distinguishing between CO2/O2 is primarily accomplished by loop 6 of the large subunit, which contains a conserved element of 11 amino acid residues. Form II enzymes are primarily anaerobic and unable to function in aerobic environments whereas form I enzymes can function in aerobic environments [59, 60].

4 μg/ml ChA21 for 72 h. Then, the coverslips were taken out, washed, fixed, and stained according to the instruction manual of in situ cell-death detection kits (Roche). The tissue sections from nude mice xenografts were dewaxed and hydrated, and then were incubated with 20 μg/L proteinase K at room temperature for 15 min, followed by incubation with TUNEL reaction mixture. Converter-peroxidase solution was added for further

incubation. Labeled nuclei were demonstrated using 3, 3′-diaminobenzidine and counterstained with hematoxylin. Four equal-sized fields were randomly chosen and analyzed, the apoptotic index (AI) was defined as follows: AI (%) = 100 × apoptotic cells/total tumor cells. Propidium iodide staining of dead cells for flow cytometry SK-OV-3 cells were incubated #Selleckchem ICG-001 randurls[1|1|,|CHEM1|]# with ChA21 (0.2 or 5.4 μg/ml) for 72 h, harvested and counted, and 1 × 106 cells were resuspended in 100 μl phosphate-buffered saline (PBS). buy Tipifarnib Then, 5 μl of propidium iodide (PI, Beckman, USA) was added, incubated for 30 min at

room temperature in dark. Then the cells were subjected to flow cytometry to measure the death rate (%) with a Beckman Coulter Epics-XL-MCL cytometer (California, USA). Immunohistochemical and immunocytochemical staining for Bcl-2 and Bax The SK-OV-3 cells were cultured and fixed as described above in TUNEL staining. The sections of paraffin-embedded tissue were dewaxed and rehydrated. After inactivating endogenous peroxidase with 3% H202, and blocking cross-reactivity with normal serum, the sections were incubated overnight at 4°C with the Bcl-2 antibody (1:150, Santa Cruz, California, USA) and the Bax antibody (1:150, Santa Cruz, California, USA), respectively. Then, below the sections were treated with streptoavidin-peroxidase reagent (Zymed, USA), and the peroxidase label was demonstrated

using 3, 3′-diaminobenzidine, counterstained with hematoxylin. Omission of the primary antibody was used as negative control. The immunostained sections were examined by using an Eclipse E800 microscope (Nikon, Japan) coupled to a digital camera. The mean optical density (MOD) of microscopic images was quantitatively analyzed by Image-pro Plus 5.02 (Media Cybernetics Inc, USA). Statistical analysis Data were expressed as mean ± standard deviation ( ± s). Comparison between groups was made by the Independent Samples t-test, P < 0.05 was considered statistically significant. Results ChA21 inhibits the growth of SK-OV-3 cells in vitro and in vivo To evaluate the effect of ChA21 on cell proliferation, human ovarian cancer cells SK-OV-3 were treated with different doses (0.067-5.4 μg/ml) of ChA21 for 72 h or treated with ChA21 (5.4 μg/ml) for 24, 48, 72, 96 h, respectively. As shown in Fig. 1A, treatment of ChA21 resulted in a dose-dependent inhibition of SK-OV-3 cell proliferation by MTT assay; the growth inhibitory rates were 5.85, 10.92, 16.55, 23.87 and 35.