CD44 is a transmembrane glycoprotein,which is a family of highly specific single chain mem-brane surface glycoproteins by a single gene.CD44 is mainly involved in the specific adhesion between cells and cells,cells and matrix.CD44v6 is a member of the CD44 family,and its correlation with metastasis and prognosis of non small cell lung cancer( NSCLC) has become a hot research topic in recent years.The current research pro-gress of CD44v6 and NSCLC are reviewed in this paper.

Aim To establish an in vitro early drug cardiac tox-icity evaluation method by human embryonic stem cells derived cardiomyocytes ( hESC-CM) and real-time cell analysis Cardio (RTCA Cardio) system. Method The hESC-CM were cultured at RTCA Cardio E-Plate 96. Impedance signals from hESC-CM were analyzed for beating rate, contraction amplitude and beating rhythm irregularity to determine the optimum inoculation density and detection duration. Based on this, we used 0. 1 % DMSO to be the solvent and quinidine (0. 2, 0. 78, 3. 13, 12. 5, 50 and 100 μmol·L - 1 ) known as affecting cardiac activity to validate this method. Result The results revealed no significant changes in the cell index (CI), transient pulse patterns, beating rate and amplitude of hESC-CM. Quinidine will affect the CI and transi-ent pulse patterns of hESC-CM and decrease the beating rate and amplitude of hESC-CM when its concentration ≥3. 13 μmol · L - 1 . And this effect is concentration-dependent, the higher the concentration,the more time they need to recover beating and the more significant the beating rate and amplitude inhibition of quinidine on hESC-CM. Conclusion The method established by hESC-CM and RTCA Cardio system can detect the effect of quinidine on the contraction of hESC-CM, and this indicates that this method has the potential to be an attractive high-throughput tool for screening potential drugs in early evaluation of drug car-diotoxicity.

Human embryonic stem ceIIs(hESC),characterized by unique capacities of seIf-renewaI and differentiation into cardiomyocytes and hepatocytes,can be used in new drug screening and drug safety evaIuation processes. Cardiotoxicity and hepatotoxicity are major obstacIes to deveIopment and marketing of new drugs. hESC-derived cardiomyocytes and hepatocytes have structuraI and functionaI characteristics,which can be used for cardiotoxicity and hepatotoxicity testing in vitroand for buiIding a drug safety evaIuation system invitrothat has the advantage of short experiment cycIes,smaII dose,Iow cost and few species differences. hESC-derived cardiomyocytes and hepatocytes have broad prospects of appIication in toxicoIogy.

OBJECTlVE To establish a real-time cell analysis system ( RTCA) for early drug car-diotoxicity evaluation. METHODS An in vitro drug cardiotoxicity evaluation method was established using RTCA Cardio system and primary cultured cardiomyocytes of neonatal rats. The beating rate, am-plitude and beating rhythm irregularity ( BRl ) of cardiomyocytes were observered after antiarrhythmic drugs, such as quinidine and lidocaine were added, to assess the effect of the above method on cardio-toxicity evaluation. RESULTS RTCA Cardo E-Plate 96 was inoculated with primary cultured cardiomyo-cytes that began to beat after 24 h and beat regularly after 48 h. The stable beating was maintained for a minimum of three days. The beating of cardiomyocytes decreased rapidly from 155±5 to 0 after incuba-tion with quinidine. The beating recovered gradually after 6 h. Quinidine at 3.1μmol·L-1 caused the beat-ing rate to return to 124±16. Quinidine allowed the beating rate to return to normal when the concentra-tion was less than 100.0μmol·L-1 . The beating rate, amplitude and BRl of cardiomyocytes changed in a concentration-dependent manner when incubating with lidocaine. The higher the concentration, the more significant the inhibition of lidocaine on cardiomyocytes. CONCLUSlON The cardiotoxicity of quinidine and lidocaine can be detected accurately using RTCA Cardio system, suggesting that this system can be used in early evaluation of drug cardiotoxicity.

OBJECTIVE To evaluate whether the tumor promoting activity syste m based on the ex-pression of 22 tumor pro motion marker genes in Bhas 42 cell transformation assay can be developed to be a new screening technique for predicting tumor promoting potential of che mical.METHODS Chose the 12-O-tetradecanoylphorbol-13-acetate (TPA)as positive che mical and use the Bhas 42 cell lines as test syste m.Define the day of seeding cells as d 0.On d 4,medium in each well was changed with the medium DF5F containing 0.05 mg·L -1 TPA or 0.5 %DMSO,after treat for 24,48 and 72 hrs,collect the cell samples.Extract the RNA fro m the cell samples and detect the expression of Ccnb1 ,Rif1 , Mc m3,Chek1 ,Jun,Fosl1 ,Hells,Vegfa,St mn1 ,Prl2c3,Scarb1 ,Phex,Orm1 ,Orm2,Nup54, Slc2a1 ,Il1 rl1 ,Rad51 ap1 ,Tfrc,Ab1 ,Car13 and Pik3r5 at different ti mepoints by RT-PCR.RESULTS Co mpared to the negative group,the expression of 3 genes was increased after treat with TPA for 24 hr which are Vegfa,Il1 rl1 and Pik3r5;the expression of 12 genes was increased after treat with TPA for 48 hr which are Chek1 ,Hells,Mc m3,Rad51 ap1 ,Vegfa,Il1 rl1 ,Prl2c3,Slc2a1 ,Tfrc,Ab1 ,St mn1 and Rif1 ;the expression of 10 genes was increased after treat with TPA for 72 hr which are Chek1 ,Hells, Rad51 ap1 ,Vegfa,Il1 rl1 ,Prl2c3,Slc2a1 ,Tfrc,St mn1 and Ccnb1 .CONCLUSION The 22 tumor pro-motion marker genes showed different sensitivity to the positive control che mical at different ti mepoints after treat with TPA.The result was consistent with the pro motion activity of TPA.The 22 tumor pro motion marker genes combined with the Bhas 42 cell transformation assay can be developed to be a new screening technique for predicting tumor promoting potential of che mical.

OBJECTIVE:To establish a method for the contents determination of tanshinol,protocatechuic aldehyde,ferulic ac-id and salvianolic acid B in Yixinshu capsule. METHODS:Dual-wavelength HPLC was performed on the column of Eclipse XDB-C18 with mobile phase of 0.5%phosphoric acid-methanol-acetonitrile(gradient elution)at the flow rate of 1.0 ml/min,the de-tection wavelength was 280 nm(tanshinol,protocatechuic aldehyde,salvianolic acid B)and 320 nm(ferulic acid),column tempera-ture was 30℃and volume was 10 μl. RESULTS:The linear range of tanshinol,protocatechuic aldehyde,ferulic acid and salvianolic acid B were respectively 9-144μg/ml(r=0.999 9),0.5-8μg/ml(r=0.999 9),0.65-10.4μg/ml(r=0.999 9)and 221.25-3 540μg/ml (r=0.999 9);RSDs of precision,stability and reproducibility tests were no more than 1.90%;the average recovery was respective-ly 100.8%(RSD=1.65%,n=9),100.1%(RSD=2.87%,n=9),100.1%(RSD=3.01%,n=9) and 99.4%(RSD=2.05%,n=9). CONCLUSIONS:The method is simple and reproducible,and can be used for the quality control of Yixinshu capsule.

OBJECTIVE:To establish a method for simultaneous determination of tanshinol,protocatechuic aldehyde,paeoni-florin,ferulic acid and salvianolic acid B in Shenshao oral liquid. METHODS:RP-HPLC was performed on the column of Eclipse XDB C18 with mobile phase of 0.5%phosphoric acid-methanol-acetonitrile(gradient elution)at the flow rate of 1.0 ml/min,the de-tection wavelength was 280,230 and 320 nm,column temperature was 30 ℃ and volume was 10 μl. RESULTS:Under the chro-matographic conditions,5 kinds of components could be completely separated,the linear range of tanshinol,protocatechuic alde-hyde,paeoniflorin,ferulic acid and salvianolic acid B were respectively 24-384 μg/ml(r=0.999 9),1.25-20 μg/ml(r=0.999 9), 40.5-648 μg/ml(r=0.999 8),1.5-24 μg/ml(r=0.999 9),145-2 320 μg/ml(r=0.999 9);RSDs of precision,stability and reproduc-ibility tests were no more than 2.2%;the average recovery was respectively 100.7%(RSD=1.23%,n=9),100.0%(RSD=2.19%,n=9),99.6%(RSD=0.87%,n=9),100.3%(RSD=1.11%,n=9) and 99.3%(RSD=2.46%,n=9). CONCLUSIONS:The method is specific with good precision and reproducibility,and can be used for the content determination of 5 components in Shenshao oral liquid.

OBJECTIVE:To establish a method for simultaneous determination of danshensu,salvianolic acid B,protocatechu-ic aldehyde,paeoniflorin and ferulic acid in Jingzhi guanxin granule. METHODS:HPLC was performed on the column of Zorbax Eclipse XDB-C18 with mobile phase of acetonitrile-methanol-0.5% H3PO4(gradient elution)at a flow rate of 1.0 ml/min,the detec-tion wavelength was 280 nm(for danshensu,protocatechuic aldehyde,salvianolic acid B),230 nm(for ferulic acid)and 320 nm (for paeoniflorin),column temperature was 30 ℃,injection volume was 10 μl. RESULTS:The linear range was 1.19-478.34 μg/ml for danshensu(r=0.999 9),0.11-44.93 μg/ml for protocatechuic aldehyde(r=0.999 9),7.49-995.20 μg/ml for salvianolic acid B (r=0.999 7),0.95-379.39 μg/ml for paeoniflorin (r=0.999 9) and 0.01-3.12 μg/ml for ferulic acid (r=0.999 5);the limits of quantitation were 1.91 ng,0.36 ng,150.00 ng,2.74 ng and 0.10 ng,limit of detection were 0.96 ng,0.10 ng,45.00 ng,1.52 ng and 0.03 ng;RSDs of precision,stability and reproducibility tests were lower than 2%;recoveries were 98.06%-99.47%(RSD=0.52%,n=6),98.01%-99.49%(RSD=0.70%,n=6),98.44%-99.45%(RSD=0.37%,n=6),96.94%-100.71%(RSD=1.27%,n=6)and 95.44%-100.44%(RSD=1.90%,n=6). CONCLUSIONS:The method is simple and accurate,and suitable for the simultaneous determination of danshensu,salvianolic acid B,protocatechuic aldehyde,paeoniflorin and ferulic acid in Jingzhi guanxin granule.

AS a result of a better understanding,people pay increasing attention to the adverse effect of traditional Chinese medicine. Chansu as a rare Chinese traditional medicine has been widely used in clinical treatment and known to have cardiotoxicity. The mechanism and relationships with blood drug concentrations need to be elucidated urgently. Bufadienolides,whose chemical structure is similar to that of digoxigenin,are the major component of Chansu and also belong to cardiac glycosides. They show both excitation and inhibition effects on the heart. Myocardial cells can not only be excited to trigger tachycardia and rapid arrhythmias at a low dose,but inhibited to induce heart block and slow arrhythmias at a high dose. As the key regulator of intracellular energy and calcium ion concentration , Na+/K+ATPase can be inhibited by bufadienolides through a combination of alpha subunits of the enzyme , resulting in calcium overload and energy disorders in myocardial cells. This might be the main mechanism of cardiotoxicity. In this paper,the progress in experimental research of substances of cardiotoxicity,animal tests,enzyme and ion channel,lipid metabolism and ion homeostasis,correlations between pharma?cokinetics and cardiotoxicity are reviewed.

OBJECTlVE To investigate the action mechanism of antidepressant fluoxetine on hERG ( ether-a-go-go-related gene ) potassium channel, and the effect of protein kinase C ( PKC ) agonist phorbol-12-myristate-13-acetate ( PMA) on fluoxetine inhibition. METHODS The whole cell patch clamp technique was used to record the change in hERG potassium current ( IKr ) on HEK293 cells that stably expressed hERG potassium channel ( hERG-HEK293 steady-state cells) , which was treated with fluoxe-tine 0.01, 0.1, 1 and 10μmol·L-1 , to study the concentration-and voltage-dependence of the effects on IKr, and to observe the changes in activation, inactivation and recovery dynamics of hERG potassium channel treated with fluoxetine 1μmol·L-1 . On this basis, the effect of PMA of 1μmol·L-1 on inhibition of fluoxetine 1 μmol·L-1 was explored. RESULTS Fluoxetine 0.01, 0.1, 1 and 10 μmol·L-1 inhibited IKr on hERG-HEK293 steady-state cells in a concentration- and voltage-dependent manner. The half maximal inhibitory concentration ( lC50 ) was about 0. 8 mmol·L-1 , and the Hill coefficient was about 1. 1. Fluoxetine 1 μmol·L-1 could reduce the activation, deactivation and recovery currents of IKr and affect the activation and recovery of hERG potassium channel. After fluoxetine inhibition of IKr became stable, PMA 1 μmol·L-1 could inhibit the blocking effect of fluoxetine on hERG potassium channels. CONCLUSlON Fluoxetine has obvious inhibitory effect on IKr of hERG-HEK293 steady-state cells, but the effect could be inhibited by PKC agonist PMA.