Objective: To investigate the effects of curcumin combined with oxaliplatin on the human colon cancer cells LoVo xenografted tumor in nude mice and to explore the mechanism. Methods: Nude mice were implanted with human colon cancer LoVo cells. All tumor-bearing mice were randomly divided into four groups and treated with vehicle, 50 mg/kg curcumin, 25 mg/kg oxaliplatin, and their combination (50 mg/kg curcumin + 25 mg/kg oxaliplatin) by ip injection once every other day individually. After continuous administration of drug treatment for 11 times, the weights of nude mice were recorded, the stripping tumor weight was monitored, and the tumor volume and tumor inhibitory rates were calculated. The enucleation of eyeball for taking blood and blood routine examination were carried out and the function of liver and kidney was detected. Tumor cell cycle and apoptosis rate were assayed by flow cytometry. The pathological morphology of tumor was analyzed by HE staining. The apoptosis related gene expression was detected by RT-PCR. Results: Tumor inhibitory rates of curcumin, oxaliplatin, and curcumin + oxaliplatin groups were 59.47%, 55.49%, and 70.56%, respectively. Curcumin combination with oxaliplatin did not influence the blood system, liver, and kidneys in nude mice. Combination of curcumin and oxaliplatin could effectively inhibit the tumor growth (P < 0.05), interfere with cell cycle arresting at S and G2/M phases (P < 0.05, 0.01), and promote the expression of bax (P < 0.01) in tumor-bearing nude mice. Conclusion: Combination of curcumin and oxaliplatin could synergistically inhibit the growth of LoVo colonic xenografts in nude mice.

Objective To determine the effects of urapidil on L-type calcium current(I_(Ca-L))in rat cardiomyocytes.Methods Ventricular myocytes were isolated from SD rats of either sex(250-280g)by retrograde perfusion of the hearts via aorta with calcium-free Tyrode solution containing enzyme as described elsewhere.Rod shaped cells with clear borders and striations were selected.Eighteen cells were randomly divided into 3 groups(n =6 each):A urapidil group;B urapidil+methysergide group and C methysergide group.All the cells in the three groups were peffused first with Tyrode solution for 1 min(T_1).In group A and C cells were then peffused with Tyrode solution containing 0.4 ?mol?L~(-1) urapidil(A)or 40 nmol?L~(-1) methysergide(C)for 1 min(T_2) while in group B cells were perfused fwst with Tyrode solution containing 0.4 ?mol?L~(-1) urapidil for 1 min (T_2) then with Tyrode solution containing methysergide 40 nmol?L~(-1) for 1 min (T_3).Finally the cells were again perfused with regular Tyrode solution for 1 min(T_4)to wash out the drugs.The peak of I_(Ca-L) was recorded at T_(1-4) by means of the whole cell patch clamp technique with use of Axo patch 200B.Results In group A,B and C the peak of I_(ca-L) at T_2 was significantly lower than that at T_1 but there was no significant difference between the peak of I_(ca-L) at T_1 and T_4.In group B the peak of I_(Ca-L) at T_3 was significantly lower than that at T_2.Conclusion Urapidil inhibits L-type calcium current in rat isolated cardiomyoeytes.It's inhibitory effect may not be mediated by 5-H_(1A) receptor.

Sixteen compounds were isolated from the aerial parts of Euphorbia sikkimensis by means of various chromatographic techniques such as silica gel, Sephades LH-20 and RP-18, and their structures were elucidated as naringenin (1), kaempferol (2), quercetin (3), kaempferol-3-O-alpha-L-arabinopyranoside (4), quercetin-3-O-alpha-L-arabinopyranoside (5), quercetin-3-O-(2"-galloyl)-alpha-L-arabinopyranoside (6), 5alpha, 8alpha-epidioxy-(22E, 24R)-ergosta-6,22-dien-3beta-ol (7), stigmast-5-ene-7-one-3beta-ol (8), 3beta-hydroxy4a, 14alpha-dimethyl-5alpha-ergosta-8, 24(28)-dien-7-one(9), beta-sitosterol (10) , 10-cucurbitadienol( 1) , scopoletin(12) , ethyl gallate(13), p-hydroxybenzaldehyde (14), 3 betahydroxybenzeneethanol( 15) ,and 2,4-dihydroxy-6-methoxy-acetophenone (16) on the basis of spectroscopic data analysis. All the compounds are isolated from this plant for the first time, and compounds 1, 4-8, 15 are obtained from Euphorbia species for the first time.
Chromatography ; Euphorbia ; chemistry ; Organic Chemicals ; analysis ; isolation & purification

Objective To study the effect of survivin anfisense oligonucleotides (ASODN)combined with quercetin on proliferation, apoptosis and cell cycle of a hepatocellular carcinoma cell line SSMC-7721 cells. Methods Human hepatocellular carcinoma cell line SSMC-7721 was cultured in vitro,and cells on logarithmic growth phase were used for this experiment. Cell proliferation was measured by MTT assay. The apoptotic rate and cell cycle were examined by flow cytometer (FCM). Morphological change of apoptotic cells were observed by fluorescent microscope. The expression of survivin gene was detected by the method of immunohistochemistry staining and RT-PCR on the mRNA and protein level. Results After sealing survivin gene with ASODN, the proliferation of SSMC-7721 cells was inhibited markedly. FCM analysis showed that there appeared an obvious apoptosis peak after transfection. The inhibitory effect of combined administration of survivin ASODN and quercetin on cell proliferation was much stronger than that of the single way. The result of immunohistochemical and RT-PCR assays showed that survivin ASODN and quercetin inhibited the expression of survivin gene. Conclusion Combined survivin ASODN with quercetin significantly inhibit cell proliferation, down-regulate survivin gene expression and induce the apoptosis of SSMC-7721 cells.

The zinc-regulated transporters (ZRT), iron-regulated transporter (IRT)-like protein (ZIP) plays an important role in the growth and development of plant. In this study, a full length cDNA of ZIP encoding gene, designed as DoZIP1 (GenBank accession KJ946203), was identified from Dendrobium officinale using RT-PCR and RACE. Bioinformatics analysis showed that DoZIP1 consisted of a 1,056 bp open reading frame (ORF) encoded a 351-aa protein with a molecular weight of 37.57 kDa and an isoelectric point (pI) of 6.09. The deduced DoZIP1 protein contained the conserved ZIP domain, and its secondary structure was composed of 50.71% alpha helix, 11.11% extended strand, 36.18% random coil, and beta turn 1.99%. DoZIP1 protein exhibited a signal peptide and eight transmembrane domains, presumably locating in cell membrane. The amino acid sequence had high homology with ZIP proteins from Arabidopsis, alfalfa and rice. A phylogenetic tree analysis demonstrated that DoZIP1 was closely related to AtZIP10 and OsZIP3, and they were clustered into one clade. Real time quantitative PCR analysis demonstrated that the transcription level of DoZIP1 in D. officinale roots was the highest (4.19 fold higher than that of stems), followed by that of leaves (1.12 fold). Molecular characters of DoZIP1 will be useful for further functional determination of the gene involving in the growth and development of D. officinale.
Amino Acid Sequence ; Cloning, Molecular ; Dendrobium ; chemistry ; classification ; genetics ; metabolism ; Gene Expression Regulation, Plant ; Iron ; metabolism ; Membrane Transport Proteins ; chemistry ; genetics ; metabolism ; Molecular Sequence Data ; Phylogeny ; Plant Proteins ; chemistry ; genetics ; metabolism ; Plants ; chemistry ; classification ; genetics ; Sequence Alignment ; Zinc ; metabolism

The strain of Cunninghamella echinulata 9980 was first selected with high aminoacylase activity . In three submerged cultures, the aminoacylase activity in the mycelial cell was compared . A number of factors have effects on the resolution reaction. The results showed that, peptone culture gave the highest aminoacylase activity with 680U/g.The optium temperature,pH,and substrate concentration were 55℃, 7.0,and 0.2mol/L,respectively. The ions in the buffer lowered the activity,but the Co~(2+) in 10~(-4)~10~(-3 )mol/L was necessary for its activity.

【Objective】To explore the optimal conditions in fingerprinting (APHDF).【Methods】The human DNA fingerprints were detected by APHDF.A pair of short primers was used for amplification.The experimental conditions including template,Mg2+,deoxyribonucleotides,and parameters of cycle,were optimized.【Results】The template DNA should be abstracted freshly and the concentration should be ranged from 50～550 mg/L.The best concentration of Mg2+was 5.0 mmol/L.The deoxyribonucleotides concentration was optimal at 0.2 mmol/L.The PCR cycling parameters were as follows :The denaturing temperatures,annealing temperatures and extension temperatures were 94 ℃ and 90 ℃ for 30 s,43 ℃ and 48 ℃ for 40 s or 50 s,and 72 ℃ for 1 min or 80 s,respectively.【Conclusion】The optimal conditions of the experiment are obtained,with good reproducibility and high specificity.Therefore,this method can be widely applied in practice.

OBJECTIVETo study the influence of different drying methods on the quality of Schisandrae Chinensis Fructus and thus provide useful reference for its proper drying methods.

METHODSchisandrae Chinensis Fructus was processed by eight drying methods including vacuum freeze drying, natural drying in the shade, drying in the sun, oven drying and vacuum drying under different temperature. The contents of the functional ingredients includes chisandrin, gomisin D, gomisin J, schisandrol B, angeloylgomisin H, angeloylgomisin Q, gomisin G, schisantherin A, deoxyschisandrin, schisandrin B, schisandrin C, 5-HMF, total aids and total sugars. The main components change after drying were analyzed by HPLC, ultraviolet spectrophotometry and potentiometric titration. Principal component analysis (PCA) was carried out by SPSS software to evaluate the quality of different processed products from Schisandrae Chinensis Fructus.

RESULTAll these results are in accordance with the requirements of Chinese Pharmacopoeia published in 2010, the contents of schisandrin and total eleven lignans were the highest using vacuum drying, and 5-HMF were the lower, oven drying made little difference but with lower schisandrin and higher 5-HMF as the heat increased.

CONCLUSIONDifferent drying methods have significant influence on the quality of Schisandrae Chinensis Fructus. Oven drying under 5°C should be adopted to substitute drying in the sun according to the China Pharmacopoeia published in 2010 for Schisandrae Chinensis Fructus by comprehensive analysis of the cost, content and practicality.

Desiccation ; methods ; Drugs, Chinese Herbal ; chemistry ; Quality Control ; Schisandra ; chemistry ; Temperature

OBJECTIVETo study the effect of Zhengganfang (ZGF) drug serum in inducing differentiation of Bel-7402 human hepatocarcinoma cell (HHC) and its influence of the telomerase activity.

METHODSRats were fed with ZGF decoction to prepare the drug-serum of ZGF, and the drug-serum was used for Bel-7402 HHC culture. Serum of normal rat, was taken as negative control and retinoic acid (RA) as the positive control. Cell proliferation was detected by MTT colorimetric assay, nucleocytoplasm ratio by image assay, alpha-fetoprotein (AFP) by RIA, alkaline phosphatase (ALP) activity and gamma-glutamyl transpeptidase (gamma-GT) by dynamic colorimetric assay and telomerase activity of Bel-7402 cell by PCR-ELISA.

RESULTSDrug-serum of ZGF could inhibit the proliferation of Bel-7402 cell, markedly reduce the nucleocytoplasm ratio and the secretion of AFP, decrease gamma-GT activity, increase ALP activity and suppress the telomerase activity of Bel-7402 cell.

CONCLUSIONZGF can induce the differentiation of Bel-7402 HHC, the main mechanism maybe through the suppression of telomerase activity.

Alkaline Phosphatase ; metabolism ; Animals ; Carcinoma, Hepatocellular ; pathology ; Cell Transformation, Neoplastic ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Hepatocytes ; pathology ; Liver Neoplasms ; pathology ; Male ; Rats ; Rats, Sprague-Dawley ; Serum ; Telomerase ; metabolism ; Tretinoin ; Tumor Cells, Cultured ; alpha-Fetoproteins ; metabolism ; gamma-Glutamyltransferase ; metabolism

Objective To confirm whether in post-resuscitation myocardial dysfunction involved in myocyte apoptosis mechanism in porcine model of cardiac arrest and apoptosis index varied from different modalities of cardiopulmonary resuscitation or not.Methods A total of 22 WZSP inbred small swine were randomly (random number) divided into sham operation group (SHAM) (n =6),defibrillation first group (DF,n =8) and chest compression first group (CF,n =8).Eight minutes after ventricular fibrillation was set up,standard CPR was carried out subsequently after defibrillation in porcine models of cardiac arrest in DF group and defibrillation after standard CPR in CF group,and hemodynamics were monitored.Twentyfour hours after restoration of spontaneous circulation (ROSC),animals were sacrificed,and myocardial specimens were examined with electron microscopy,Western blot,quantitative RT-PCR,and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay.The experimental data were analyzed by SPSS 17.0.Student's t test was employed for comparisons between two groups.Differences within groups at different time intervals were compared with repeated measures ANOVA.A two-tailed value of P ＜ 0.05 was considered statistically significant.Results Myocardial function was significantly impaired after ROSC.Levels of Bcl-2,Bax and caspase-3 protein was markedly increased in the CF and DF groups than those in the SHAM group (P ＜ 0.05) at 24 h after ROSC,while Bcl-2/Bax was significantly reduced in the CF and DF group compared with the SHAM group (P ＜ 0.05),and much more apoptotic cells were observed in cardiac arrest animals in comparison with sham-operation animals (P ＜ 0.05).Six hours after ROSC,hemodynamic indicators improved significantly in group DF than those in group CF,but Bcl-2,Bax and caspase-3 protein levels and apoptotic index were not significantly different bewteen the DF group and CF group (P ＞ 0.05).Conclusions Caspase-3-mediated apoptosis might be one of the main pathological mechanisms of postresuscitation myocardial injury in a porcine model of cardiac arrest,but there was no statistically significant difference in apoptosis index between two resuscitation modalities,showing no one modality was superior over another.