Objective:To study the in vitro bacteriostasis of water extract of humulus. Methods:By using double dilution method and 96-well plates trace broth dilution method, the inhibitory test of water extract of humulus was conducted on Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa. The minimum inhibitory concentration ( MIC ) , minimum bactericidal concentration ( MBC) and the diameter of inhibition zone were determined. Results: The drug showed different sensitive degree on the test strains. MIC and MBC for Escherichia coli were 31. 25 mg·ml-1 , that for Staphylococcus aureus were 3. 91 mg·ml-1 , and that for Pseudo-monas aeruginosa was 125 and 250 mg·ml-1 . Conclusion:With the water extract of humulus as the research object, the study on the in vitro bacteriostasis effect of humulus scandens is closer to the clinical application form, which provides reference for the further re-search on the related preparation of humulus scandens.

Objective:To optimize the extraction process of antitumor active ingredients in fructus ligustri lucidi. Methods:Using fructus ligustri lucidi medicinal herb as the raw material, the extraction process of luteolin and pelargidenon in the herb was screened by D-optimal design. The extraction yield of different extraction processes was compared to obtain the optimal extraction conditions. Re-sults:The best extraction conditions were as follows:adding 12-fold 95% ethanol, extracting 2 times with 60 minutes for each time. Conclusion:The optimal extraction technology can provide significant reference for the further pharmacodynamic effect development of fructus ligustri lucidi.

Objective To investigate the effects of hypoxia-inducible factor (HIF)-1α on glycolysis of human esophageal squamous carcinoma cells and the possible mechanism.Methods TE13 and Eca109 cells were cultured under normal oxygen (20％O2) and hypoxia (1％O2) conditions.The hypoxia was duration 6 hours,12 hours,24 hours and 48 hours.HIF-1α gene was stable silented by RNA interference method and TE13/small interfering HIF cells and Eca109/siHIF cells were obtained.The cell culture condition and time was same as TE13 and Eca109 cells.The changes of HIF-1α expression were detected by Western-blot.The changes of lactic acid concentration in cell culture supernatant were determined by Spectrophotometry.The changes of glucose transporter-1 (GLUT-1) and lactic dehydrogenase A (LDHA) expression at mRNA level were examined by realtime polymerase chain reaction.The changes of GLUT-1 and LDHA expression at protein level were tested by Western blot.Using t or t' test to analyze the effects of hypoxia duration on HIF-1αexpression at protein level.One-way ANOVA was applied for the difference analysis between the groups.Results In TE13 and Eca109 cells,the HIF-1α expression significantly increased under hypoxia condition and reached the peak at 12 hour (t=6.11,8.31; both P＜0.05).The lactic acid secretion of TE13/siHIF cells and Eca109/siHIF cells was (1.24±0.33) and (1.28±0.37) mmol/L,which significantly decreased when compared with TE13 and Eca109 cells [(3.25±1.34) and (4.91±1.69) mmol/L,t=2.53,3.59,both P＜0.05].The lactic acid secretion of TE13 and Eca109 cells significantly increased after hypoxia [(6.48±1.73) and (8.02± 1.95) mmol/L,t=2.715,2.050,both P＜0.05].There was no significant lactic acid secretion in TE13/siHIF cells and Eca109/siHIF cells after hypoxia (P ＞ 0.05).The expressions of GLUT-1 and LDHA at mRNA level were significantly suppressed in TE13/siHIF cells and Eca109/siHIF cells (normal oxygen:t=6.98,3.92,7.25,3.67,all P＜0.05).The expression of GLUT-1 at protein level remarkably weaked (normal oxygen:t=4.57、16.56,hypoxia:t=6.19、6.09,all P＜0.05),while the expression of LDHA at protein level slightly decreased (P＞0.05).Conclusions The level of glycolysis can be lowered by suppression HIF-1α expression in human esophageal squamous carcinoma cells.The pathway may be involved in the suppression of GLUT-1 and LDHA expression.Except for HIF-1α,there may be other regulating factors in LDHA protein expression at same time.

Objective:To investigate the volatile constituents in Folium isatidis. Methods:The volatile constituents from Folium isatidis were analyzed by head-space solid micro-extraction coupled with GC-MS for the first time. Results: Thirty-five compounds (89. 95%) were identified from the volatile constituents in Folium isatidis. The main volatile constituents of Folium isatidis were 6, 10, 14-trimethyl-2-pentadecanone (6. 32%), nonanal (5. 99%), phenethyl isothiocyanate (5. 79%) and palmitic acid (5. 62%). Conclusion:Palmitic acid and benzyl alcohol may be the main effective constituents in Folium isatidis.

BACKGROUND:5-Fluorouracil occupies an important position in the treatment of gastric cancer, but its long-term use can easily induce adverse reactions such as myelosuppression and leukopenia. Polylactic acid and its copolymers have a higher biocompatibility, and their decomposer cannot gather in the body.
OBJECTIVE:To investigate the in vitro cytotoxicity mechanism of 5-fluorouracil-loaded polylactic acid nanoparticles on gastric cancer cel lines.
METHODS:Ten mice were selected in this study. 5-fluorouracil-loaded polylactic acid nanoparticles (1×10-7, 1×10-6, 1×10-5, 1×10-4 mol/L) were prepared using ultrasonic emulsification method. Kiling effect of polylactic acid nanoparticles on gastric cancer cel lines in vitrowere detected. Then, the inhibition rate was calculated at different concentrations.
RESULTS AND CONCLUSIONS: Under the transmission electron microscope, 5-fluorouracil-loaded polylactic acid nanoparticles had good shape and relatively evenly distributed with no adhesions. After drug administration, the drug concentration was 50% at 24 hours and 62.9% at 72 hours. After 48 hours co-culture with single 5-fluorouracil or 5-fluorouracil-loaded polylactic acid nanoparticles, the viability of gastric cancer cels showed a decrease trend with the increase of drug concentrations, and moreover, 5-fluorouracil-loaded polylactic acid nanoparticles had a better cel inhibition ability than the single 5-fluorouracil (P < 0.05). The IC50value of 5-fluorouracil-loaded polylactic acid nanoparticles was significantly lower than that of 5-fluorouracil (P < 0.05). These findings indicate that polylactic acid nanoparticles as good drug carriers have a strong drug loading capacity and increase drug concentration in the body, but cannot reduce the biological activity of 5-fluorouracil, which provide new ideas for the treatment of gastric cancer.

Objective:To compare the content of icariin in Rujiling granules before and after the ray irradiation. Methods: The column was Thermo ODS-2 HYPERSIL (250 mm × 4. 6 mm,5 μm) with the temperature of 30℃, and the mobile phase was acetoni-trile-0. 033 mol·L-1 potassium dihydrogen phosphate(30∶70) with the flow rate of 1. 0 ml·min-1. The detection wavelength was 265nm. Results:There was a good linear relationship within the range of 3. 44-34. 40μg·ml-1 for icariin (r=0. 999 7, n=6). The average recovery was 98. 18% with RSD of 0. 32%(n=9). Conclusion:The content change of icariin in Rujiling granules before and after the ray irradiation is not statistically significant, which illustrates that the conventional dose irradiation has little effect on icariin.

Objective:To comparatively analyze the effect of different methods on the content determination of baicalin in Qing-houyan mixtures. Methods:The content of baicalin in Qinghouyan mixtures was quantitatively determined by orthogonal function spec-trophotometry and HPLC method, respectively. Results: The regression equation of the proposed method was obtained as follows:P2 =0. 091 7C-0. 005 9(r=0. 999 9). The linear relationship was good within the range of 0. 045-0. 225 mg/ml. The determina-tion results had no statistical difference from those of HPLC(P>0. 05). Conclusion:The orthogonal function spectrophotometry meth-od is simple, fast and efficient, which can be applied in the determination of the main active components in traditional Chinese medi-cine preparations.

Objective:To study the chemical constituents in Blumea aromatica ( Wall. ) DC. Methods:The compounds were iso-lated by silica gel, Sephadex LH-20 column chromatography and TLC. The structures were elucidated by physicochemical properties and spectral analysis. Results:Ten compounds were obtained and elucidated as oleic acid (1), linoleic acid (2), quercetin (3), luteolin (4), naringenin (5), hesperetin (6), gallic acid (7), β-sitosterol (8), daucosterol (9) and hesperidin (10). Conclu-sion:Compounds 1, 2, 5, 6, 7 and 10 are isolated from the plant for the first time.

Objective To investigate the changes of hypoxia-inducible factor(HIF)-1α and hexokinase-Ⅱ(HK-Ⅱ)expression in human esophageal squamous cell carcinoma and its effect in glycolysis.Methods TE13 cells and Eca109 cells were cultured under hypoxic condition(1 ％O2)for different hypoxic time(6,12,24 and 48 hours).Cells cultured under normal oxygen condition(20％O2)were set as control.The changes of HIF-1α and HK-Ⅱ expressions at protein level were detected by Western blot.HIF-1α genes were specifically silenced with RNA interference technology(RNAi),and then the changes of HIF-1α and HK-Ⅱ expression were determined by realtime PCR and Western blot.Under normal oxygen and hypoxic condition,the changes of lactic acid concentration in cell culture medium were detected by spectrophotometric method.Results Under hypoxic condition,the expression of HIF-1α and HK-Ⅱ gradually increased as hypoxic time extended(P＜0.05),reached a peak at 12h and then gradually decreased as time extended.Compared with that under normal oxygen condition,the expression of HK-Ⅱ in TE13 cells and Eca109 cells significantly increased under hypoxic condition(P＜0.05),which was more significant after 12 hours hypoxia.The result of realtime PCR indicated that under normal oxygen condition the expression of HIF-1α at RNA level in TE13/shRNA cells and Eca109/shRNA cells significantly decreased compared with TE13 cells and Eca109 cells without interference(P＜0.05).The expression of HK-Ⅱ at RNA level was consistent with the result of HIF-1α.Under normal and hypoxia condition,the expression of HK-Ⅱ at protein level in TE13/shRNA cells and Eca109/shRNA cells significantly decreased compared with TE13 cells and Eca109 cells without interference,and the difference was statistic significant(P＜0.05).The lactic acid secretion of TE13 cells and Eca109 cells under hypoxia condition(14.707 ± 3.594 and 15.062 ±3.901)was higher than that under normal oxygen condition(6.070±1.839 and 6.891±1.592,P＜0.05).The lactic acid secretion of TE13/shRNA cells and Eca109/shRNA cells significantly decreased compared with TE13 cells and Eca109 cells without interference,and the difference was statistic significant(P＜0.05).Conclusion The expressions of HIF-1α and HK-Ⅱ in human esophageal squamous cell carcinoma significantly increased under hypoxia conditions.The expression of HK-Ⅱ is closely correlated with lactic acid concentration and HIF-1α expression.HIF-1α may affect cell glycolysis through HK-Ⅱ.

Objective To investigate the inhibitory effect of blocking PI3K/AKT pathway by wortmannin on hypoxia-inducible factor 1α (HIF-1α) and the effect on the expression of glycolysis associated genes in human esophageal carcinoma cell lines TE1 and TE13,and to analyze the relation between PI3K/AKT-HIF-1α pathway and glycolysis in esophageal carcinoma cells. Methods Esophageal carcinoma cell lines TE1 and TE13 pretreated with wortmannin (2 μmol/L) were incubated under normoxic and hypoxic conditions.And each cell line was divided into four groups.The expression of HIF-1α and glycolysis associated genes GLUT-1,LDHA and HK-Ⅱ at protein level were measured by.Western blot.The expression of HIF-1α,GLUT-1,LDHA and HK-Ⅱ at mRNA level was determined by real-time PCR. The activities of LDH and HK-Ⅱ and lactic acid (LA)concentration in the culture supernatant were tested with spectrophotometer method.Results Under normoxic condition,HIF-1α was expressed in TE1 cells and the expression of HIF-1α was inhibited by wortmannin (2 μmol/L),the most significant inhibitory effect was at 12 hours,therefore 12 hours was selected for the subsequent hypoxia experiment.Compared with untreated cells,the expression of HIF-1α、HK-Ⅱ 、GLUT-1、LDH-A at protein level significantly decreased in TE1 and TE13 cells after pretreated with wortmannin (P ＜ 0.05),and the expression of HIF-1α、HK-Ⅱ at mRNA level significantly decreased (P＜ 0.05).Under normoxic and hypoxic conditions,the HK-Ⅱ and LDH activities in TE1 and Te13 esophageal carcinoma cells significantly decreased after treated with wortmannin compared with untreated cells (P＜0.05).Under hypoxia condition,the enzyme activity increased in untreated cells (P＜ 0.05). Under normoxic and hyp0xic conditions,the lactic acid concentration in the culture supernatant obviously decreased in cells treated with wortmannin compared with untreated cells (P＜ 0.05). Under hypoxia condition,lactic acid concentration increased in wortmannin treated cells (P ＜ 0.05). Conclusions Under normoxic and hypoxic conditions,wortmannin decrease lactic acid concentration through inhibiting the expression of HIF-1α and glycolysis associated genes, which indicate PI3K/AKT-HIF-1α pathway was closely related to glycolysis in esophageal carcinoma cells.