OBJECTIVE To evaluate the anti-tumor activities of WX-127-07,a new microtubule-tar-geting agent invitroand probe its molecular mechanism. METHODS The well-known microtubule-targe-ting anti-tumor drugs taxol,vincristine and anti-gout drug colchicine were used as positive controls. The anti-proliferation activity was examined in five different cell lines after treatment with WX-127-07(0.3 -300 nmol·L-1 )for 72 h by SRB assay. The cell cycle arrest profile was assayed by flow cytometry. The multiparameters of cytotoxicity,cell morphology,apoptosis and different signaling pathways related to tumorigenesis and inflammation were analyzed using the high content analysis platform. Tubulin tryptic digestion and competition inhibition assay for colchicine or vinblastine site were used to confirm the bind-ing site in microtubules at a molecular level. RESULTS All the tested compounds obviously inhibited the growth of A549,HepG2,HeLa,HLF and HUVEC cells. The lC50 values of WX-127-07 were 4.47±0.05, 5.18±0.08,4.90±0.19,4.10±0.16 and(5.04±0.08)nmol·L-1 respectively,lower than those of colchicine〔the lC50 values were 21. 17 ± 1. 22,14. 19 ± 0. 53,43. 80 ± 1. 64,145. 89 ± 10. 97 and( 27. 67 ± 1.79)nmol·L-1 ,respectively〕and those of vincristine〔the lC50 values were 16.51±0.36,16.76±0.33, 27.80±2.75,43.80±1.48 and(9.15±0.78)nmol·L-1 ,respectively〕,but were similar to or lower than those of taxol〔the lC50 values were 10. 68 ± 0. 61,12. 86 ± 0. 25,4. 81 ± 0. 61,102. 07 ± 15. 17 and( 3. 04 ± 0.12)nmol·L-1 ,respectively〕. High content multi-parameter analysis revealed that WX-127-07 induced a concentration-dependent microtubular depolymerization(P=0.0075)with the same pattern as colchicine and vincristine,but at a lower concentration. Both WX-127-07 and positive drugs could induce cell cycle arrest in A549 cells,increase nuclear membrane permeability and early signs of apoptosis in HepG2 cells,but neither cancer related pathways nor inflammation related pathways were affected. Microtubular competition inhibition assay showed that WX-127-07 inhibited the binding of colchicine with tubulin(P =0.0259). Tryptic digestion of tubulin-WX-127-07 premixture showed a similar electrophoretic band to that of tubulin-colchicine premixture. CONCLUSION WX-127-07 is a novel microtubule-depolymerizing agent with anti-proliferation activity and acting on the colchicine binding site.

OBJECTIVE To investigate the protective effect of velvet antler polypeptides(VAP)on hydrogen peroxide( H2 O2 )-induced injury in vascular endothelial cells and the possible mechanism.METHODS The EVC-304 cells cultured invitrowere incubated with H2 O2 for another 12 h after pretreat-ment with VAP 20,40 and 80 mg·L-1 for 24 h. Cell viability was determined by MTT assay, Hoechst333258 staining was used to observe cell morphology,the activity of superoxide dismutase (SOD)and the level of malondialdehyde( MDA)were detected with kits and the expression of heat shock protein(HSP70)and caspase 3 was detected by Western blotting. RESULTS Compared with the normal control group,the cell survival rate was decreased significantly in H2 O2 injury group( P ﹤0.01),cell shrinkage,chromatin condensation,and nuclear fragmentation were seen,the intracellular SOD activity decreased while MDA content increased(P﹤0.01),and caspase 3 and HSP70 expression increased(P﹤0.01). Compared with H2 O2 group,the cell survival rate in VAP 20,40 and 80 mg·L-1 pre-treatment groups increased significantly(P﹤0.01),the apoptosis ratio declined from(25.3±1.0)% to (15.2±1.2)%,(10.3±0.9)% and(7.9±1.4)%(P﹤0.01),the SOD activity increased to 19.2±0.5,22.3± 1.7 and(24.9±0.6)kU·g-1 protein(P﹤0.01),MDA concentration decreased to 1.51±0.2,1.48±0.3 and (1.02±0.1)μmol·g-1 protein(P﹤0.01),and the expression of caspase 3 and HSP70 declined significant-ly(P﹤0.01). CONCLUSION VAP has exert protective effect on H2 O2-induced injury in vascular endothe-lial cells. The possible mechanism might be related to improvement of intracellular oxidative stress level.

OBJECTlVE To develop an LC-MS/MS method for simultaneous determination of pro-damine ( PDM) and its metabolite 2,4-dinitro-N3-propyl-6-trifluoromethyl-1,3-benzenediamine ( DTB) in rat plasma in order to study toxicokinetics of PDM in rats. METHODS SD male rats were administered a single dose of PDM ( ig: 100 and 1000 mg·kg-1; iv: 100 mg·kg-1 ) . LC-MS/MS method was used to determine PDM and DTB in rat plasma. Toxicokinetic parameters were fitted using DAS Ver2. 1. 1. RESULTS After ig administration of PDM 100 mg·kg-1 , the parameters of PDM and DTB were as fol-lows:AUC(0-t) was 2715±102 and (6845±316)μg·h·L-1, t1/2z was 9.0±1.4 and (7.1±1.3)h, Tmax was 7.0± 1.6 and (7.0±0.0)h, cmax was 146±51 and (473±103)μg·L-1. After ig administration of PDM 1000 mg·kg-1, the parameters of PDM and DTB were as follows:AUC(0-t) was 3401±242 and (10364± 573)μg·h·L-1, t1/2z was 8.8±2.1 and (6.0±1.8)h, Tmax was (7.0±1.6)h, cmax was 175±56 and (586± 152)μg·L-1 . The absolute bioavailability of PDM was 44.9%( 100 mg·kg-1 ) and 17.1%( 1000 mg·kg-1 ) . CONCLUSlON This method is suitable for the analysis of PDM and DTB in rat plasma. There is evidence that PDM and DTB display nonlinear toxicokinetic characteristics in the studied dose range.

OBJECTIVE To explore the effect and underlying mechanism of promethazine(PMZ) on proarrhythmia in guinea pigs. METHODS ① InvivoECG recordings were made to analyze effects of jugular intravenous(iv)injection of PMZ on ECG in guinea pigs. PMZ was injected in this order:3.83→7.67→15.33→38.33 mg·kg-1 cumulatively. ② In vitroECG recordings were made to analyze effects of PMZ on ECG in isolated hearts of guinea pigs. PMZ was perfused in such order:0. 1 → 1 → 10 →50 μmol·L-1 . ③ L-type Ca2+ currents from ventricular myocytes in guinea pigs were recorded to investi-gate the PMZ's blocking effect. PMZ was perfused in such order:0.1→1→10→50 μmol·L-1→washout.④ hNav1.5 and hERG currents were recorded to investigate the PMZ's blocking effects. PMZ-perfused in such order:1→3→10→30 μmol·L-1 for hNav1.5 current analysis,and 0.3→1→3→10 μmol·L-1 for hERG current analysis. RESULTS ① PMZ(15.33 mg·kg-1 )significantly prolonged QRS intervals in guinea pigs invivoECG(P﹤0.05). PMZ(38.33 mg·kg-1 )prolonged QRS,QTc,and P-R intervals but reduced the heart rate( P﹤0.05). PMZ(10 μmol·L-1 )reduced the heart rate of isolated guinea pig hearts. PMZ 50 μmol·L-1 prolonged QRS and QTc intervals and further reduced the heart rate(P﹤0.05).③ PMZ inhibited the L-type Ca2+ current from ventricular myocytes in guinea pigs in a concentration-dependent manner with the lC50 of(8.9±1.0)μmol·L-1 . ④ PMZ inhibited the hNav1.5 and hERG currents in a concentration-dependent manner with the lC50 of 6.1±1.5 and(1.6±0.2)μmol·L-1 ,respectively. CONCLUSION PMZ might cause arrhythmia at overdoses and incombination with other drugs which have potential blocking effect on /Na ,Ca2+ and /kr currents. The proarrhythmic effect of PMZ might be mediated by the blocking effect on /Na ,Ca2+ and /kr currents.

OBJECTIVETo investigate the effect of ganoderma lucidum polysaccharides on blood lipid and lipoperoxidation from the experimental hyperlipidemic rats.

METHOD50 rats were randomly divided into normal group, hyperlipidemia control group, experimental group 1, 2 and 3 in which the rats were treated with ganoderma lucidum polysaccharides at dosages of 200 mg x kg(-1) and 400 mg x kg(-1) and 800 mg x kg(-1) respectively. Apart from the rats in control group, all the rats in other groups were fed with high fat forage for 30 days. The blood was collected from the tails of rats for measuring the serum TC, TG, HDL-C, LDL-C, GSH-Px, SOD and LPO.

RESULTGanoderma lucidum polysaccharides could significantly decrease the serum contents of TC, TG, LDL-c in the experimental hyperlipidemic rats (P < 0.01), and markedly increase the level of serum HDL-C (P < 0.05), Mean Level of blood LPO in the experimental groups treated by ganoderma lacidum polysaccharides at different dosages were much lower than that in hyper lipidema group, and the GSH-Px and SOD activities of blood in the group of ganoderma were much higher than those in hyperlipidema group.

CONCLUSIONGanoderma can regulate lipid metabolism, enhance the antioxidation and reduce the lipid peroxidation in the rats with hyperlipidemia.

Animals ; Cholesterol, HDL ; blood ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Hyperlipidemias ; drug therapy ; Hypolipidemic Agents ; pharmacology ; therapeutic use ; Lipid Peroxidation ; drug effects ; Lipids ; blood ; Male ; Phytotherapy ; Polysaccharides ; isolation & purification ; pharmacology ; therapeutic use ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reishi ; chemistry

Brain natriuretic peptide (BNP) is a major marker for evaluating cardiac function and has been widely used in clinical practice. Recent researches show that BNP is also useful for identification of sudden cardiac death in forensic pathology. This article reviews the molecular structure and biological characteristics of the BNP and its application as a functional indicate in forensic medicine. It shows that the expression of BNP in cardiac muscles, together with the expression of BNP in blood and pericardium liquid can be used to evaluate the pathological physiology changes and dysfunction degrees of the heart during the cardiac sudden death.
Amino Acid Sequence ; Animals ; Autopsy ; Biomarkers ; Death, Sudden, Cardiac ; Forensic Pathology ; Heart Diseases/physiopathology* ; Heart Failure/physiopathology* ; Humans ; Immunohistochemistry ; Molecular Sequence Data ; Myocardium/pathology* ; Natriuretic Peptide, Brain/metabolism* ; Peptide Fragments/metabolism* ; Pericardium/metabolism* ; Postmortem Changes ; RNA, Messenger/metabolism*