Aim To introduce a high throughput screen model for PARP-1 inhibitors. Methods Setting up an assay for PARP-1 activity relies on the conversion of NAD~+into a highly fluorescent compound. The inhibitory effects of 9 280 samples(including pure organic compounds, extracts from plants and extracts from microorganism)were screened by the high throughout assay. Results 148 compounds had inhibitory effects over 70%. Ultimately, three inhibitors were identified as PARP-1 inhibitor with high activity.Conclusion The high throughput screening was a highly sensitive, inexpensive, and operationally simple assay method in identifying PARP-1 inhibitors.

Objective To observe the autophagy of 87-MG and U251 glioma cells induced by ceramide and explore the possible mechanism. Methods The viability and apoptosis of 87-MG and U251 cells were detected by MTT assay and flow cytometry, respectively. Autophagic- related protein expressions of LC3B /LC3A and Beclin-1 were determined by Western blotting. The activation of JNK/c-Jun signaling pathway induced by ceramide with or without the treatment of JNK specific inhibitor SP600125 was also measured. Results 24 hours after treatment of ceramide, the growth of 87-MG and U251 cells was significantly inhibited time-dependently (P<0.05); and the number of autophagic cells increased dose-dependently (P<0.05). The levels of LC3B/LC3A and Beclin-1 significantly increased after ceramide treatment (P<0.05). JNK signaling pathway was activated in the 87-MG and U251 cells and the phosphorylation of c-Jun also increased after ceramide treatment. This activation of autophagy could be reversed by the pre-treatment of SP600125. Conclusion Ceramide may induce autophagy in 87-MG and U251 glioma cells and the mechanism may be related to the activation of JNK/c-Jun signaling pathway.

The effects of DHAA-urea,a novel dehydroabietylamine(DHAA) derivatives,on cell viability and glucose metabolism,in hypoxia and normoxia human hepatoma HepG2 cells were investigated.Hypoxia cells were achieved using DMEM containing high concentration of glucose without serum and pre-incubating of CoCl_2 (final concentration 150 μmol/L) for 24 h.The antiproliferation effect of DHAA-urea was measured by colorimetric MTT assay.The cellular ATP concentration,the lactate dehydrogenase(LDH) and glucose-6-phosphate dehydro genase (G6PD) activity were detected by their kits.It was shown that DHAA-urea markedly inhibited cell viability,cellular ATP level,LDH and G6PD activity in either aerobic or anaerobic circumstance in a dose-and time dependent manner.This suggested that DHAA-urea possibly inhibited HepG2 cells growth via the inhibition of glucolysis and glucolysis-dependent ATP depletion.DHAA-urea could be a promising candidate in the development of a novel class of agents used for human hepatocellular carcinoma.

Objective To analyze and compare results of local lymph node assays (LLNA) between BALB/c and CBA mice.Methods 4 chemicals (2,4-dinitrochlorobenzene, eugenol, hexyl cinnamic aldehyde and isopropanol) and 3 cosmetics were applied to the dorsum of both ears of Balb/c and CBA mice for three consecutive days.BrdU solution was injected inter-peritoneally on day 5.On day 6, the bilateral draining auricular lymph nodes were excised and made into single cell suspension, the lymph cell proliferation was measured by BrdU ELISA kit.Results 2,4-dinitrochlorobenzene, eugenol, hexyl cinnamic aldehyde and NO.3 perm agent pretended positive for both strains, EC1.6 values of three chemicals were found to be 0.08% (very strong), 4.02% (moderate), 6.68% (moderate) and 0.07% (very strong), 6.08% ( moderate), 8.89% ( moderate) for BALB/c and CBA mice respectively.Isopropanol, NO.1 and NO.2 cosmetics pretended to be non-sensitizers with SI <1.6 for both strains.Conclusion This study showed that BALB/c mouse was essentially equal to CBA for LLNA: BrdU-ELISA, which suggested that BALB/c mouse was a good alternative for CBA used in chemicals and cosmetics allergenic evaluation.

Object To study the hemolytic effect of total saponins from Ginseng (Gs), Radix Ophiopogonis (ROs) and Shengmaisan, a compound preparation of the two saponins, and the antihemolytic effect of antioxidants on hemolysis. Methods The percentage of hemolysis was indirectly calculated by measured cyanomethemoglobin, and morphological changes of red blood cells (RBCs) was examined by light microscope. Results Gs did not lead to hemolysis while both ROs and Shengmaisan induced lysis in a concentration dependent manner. RBCs deformation can be observed by light microscopic study. The hemolysis was partly inhibited by addition of glucose or antioxidants including mannitol and ?-tocopherol. Conclusion The intensity of hemolytic effect varied with different saponins present in the compound preparation, and may be associated with the type of saponin and partly due to its ability to enhance lipid preoxidation of RBCs membrane.

Aim The effect of N-arachidonoyl ethanolamide(Anandamide,AEA),on the human lung cancer cell A549 was analysed the possible mechanism was detected.Methods To assess the sensitivity of A549 to AEA,A549 cells were exposed to increasing doses of AEA with or without the antagonists to vanilloid receptor 1(VR1) and aspirin.MTT methods were employed to investigate A549 cell proliferation.Results A549 cells exhibited dose-dependent sensitivity to AEA resulting in dramatic cell death.But the effect of AEA on A549 could not be antagonized by the antagonists such as capsazepine and Ruthenium Red.However,cyclooxygenase(COX) inhibitor,aspirin,could attenuate A549 cell death caused by AEA(P

Glioma-infiltrating lymphocytes (GIL) were isolated from 9 surgical biopsy specimens of primary brain gliomas using mechanical and enzymatic digestion and discontinuous density gtadient centrifugation. During cultured in the presence of interieukin-2 (IL-2) for a period of four weeks, GIL were expanded 48.4-fold on the averags, even up to 118-fold. GIL activated by IL-2 had specific cytorytic activity against autologous glioma cells. Analysis of T subsets of GIL freshly isolated showed that CD3+ cells were 71.0?11.9%, CD4+ cells 34.2?6.1% and CD8+ cells 37.0?7.6%. Ability of activated GIL to secrete ?-interferon (?-IFN) was significantly higher than that of freshly isolated GIL and autologous peripheral blood lymphocytes (PBL). The results suggest that GIL have many advantages for an adoptive immunotherapy of patients with brain gliomas and is a new type of antitumor immune effector.

Aim: To search for novel modulators of glycogen metabolism through structural modifications of natural pentacyclic triterpenes. Methods: A series of N-heterocyclic derivatives were synthesized by fusing indole, qui-noxaline and pyrazine rings with A-ring of oleanolic and ursolic acids. The compounds were biologically evaluated for their inhibitory activity against rabbit muscle glycogen phosphorylase. Results and Conclusion: Twelve heter-ocyclic triterpene derivatives were synthesized and their structures were confirmed by IR, ~1H NMR, ~(13)C NMR and MS. Except for compound 12, all of the compounds exhibited glycogen phosphorylase inhibitory activity with IC_(50) values in the range of 14-252 μmol/L Among this series of compounds, compound 15 showed the best potency with IC_(50), of 14 μmol/L

Purpose A RP-HPLC method was used to determine genioside and breviscapine in plasma and to study its pharmacokinetics in rat, respectively.Methods Rat plasma samples were collected after a single dose of Zhideng injection and pharmacokinetic parameters of genioside and breviscapine were estimated,respectively.Results A good linear relationship was obtained between 0.2-40.0 μg/mL for breviscapine, and 0.5-200.0 μg/mL for genioside.The recoveries from plasma were larger than 85%,and RSDs of inter-day asaay and intra-day assay were below 10%. The pharmacokinetic results showed that genioside and breviscapine were rapidly eliminated from plasma after iv administration of three doses of Zhideng injection.The mean half-life was 72.6 min and 21.6 min,respectively.Conclusion The established HPLC method was suitable for the pharmacokinetic study of genioside and breviscapine.

To observe the effects of emodin ( Emo) on acute fatty liver in mice induced by DL-ethionine ( DL-Eth) or tetracyclin ( Tetra) and its potential mechanism, ICR mice of acute fatty liver model induced by DL-Eth were orally administered with Emo or positive control, ursodeoxycholic acid ( UDCA) for 7 days. On day 7, except that the control and Emo groups were treated with vehicle control, animals were orally administered with DL-Eth to induce acute fatty liver model. ICR mice of acute fatty liver model induced by Tetra were orally administered with Emo or positive control, Dong Bao Gan Tai ( DB) or total flavonoid C-glycosides from Abrus mollis extract ( AME) for 7 days. From day 4, except that the control group was treated with vehicle control, animals were injec-ted with Tetra intraperitoneally for 4 days to induce acute fatty liver model. Liver histopathological analyses were determined by HE staining. Serum aspartate transaminase ( AST) , alanine transaminase ( ALT) , serum triglyceride ( TG) , hepatic TG and hepatic total cholesteol ( TC) were detected. The expression of phosphorylated AMP-activa-ted kinase ( p-AMPK) , phosphorylated acetyl-CoA carboxylase ( p-ACC) , SREBP1 and fatty acid synthase ( FAS) were determined by Western blot. The expression of fatty acid translocase ( CD36 ) , peroxisome proliferator activa-ted receptor alpha ( PPARα) and microsomal triglyceride transfer protein ( MTTP ) in liver were determined by RT-PCR. Compared with model groups, Emo could improve hepatocyte swelling and hepatic steatosis induced by DL-Eth or Tetra. Serum AST, ALT, serum TG, hepatic TG and hepatic TC were decreased by Emo significantly. DL-Eth-induced increase of fatty acid synthetase-associated protein was down-regulated by Emo. Fatty acid uptake was down-regulated by Emo; fatty acid oxidation and secretion were increased by Emo. Emo might be effective in preventing acute fatty liver in mice induced by DL-Eth or Tetra.