Objective:To study the effect of GABA and A receptor agonist THIP on proliferation and apoptosis of cytokine-induced killer (CIK) cells.Methods:CIK cells were cultured by routine method,and then treated with different concentrations of GABA and THIP.The proliferation of CIK cells were investigated by MTT assay.The apoptosis,cell cycle and immunophenotype were investigated by flow cytometry.Results:GABA could inhibit the proliferation of CIK cells in a dose-dependent manner and affect the distribution of cell cycle of CIK cells(P

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.

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.

It is a thorny problem of modern medicine that the in-stent restenosis (ISR) after percutaneous coronary intervention (PCI). Combining with the etiology and pathogenesis of TCM, Professor LIU Zhong-yong believes that the root cause of ISR after PCI is the deficiency syndrome: menstruation gradually dying up, the heart yang qi deficiency; and the direct cause is excess syndrome: endogenous turbidity syndrome, heart vessel blockage. The cause for the formation of turbidity syndrome is cold, phlegm, blood stasis, poison, and dampness. Depending on the clinical manifestations, five kinds of syndromes were divided: cold turbidity stagnation, phlegm turbidity resistance, blood stasis blockage veins, poison turbidity, and dampness turbidity resistance. Professor LIU Zhong-yong also proposed relevant treatment for both symptoms and root causes, which provided new ideas and experience in the integrated TCM and Western medicine for ISR after PCI.

Objective To explore the effect of a tobacco control manual for smokers in communities of Changsha.Methods 159 qualified smokers were selected from 6 communities of Yuelu District by Multistage cluster sampling.According to their different locations.the samples were divided into two groups:77 in intervention group and 82 in control group.4 times in assessment were carried out for smokers in two groups at baseline(V0),1 month after intervention(V 1),3 months after intervention (end,V3) and follow-up(6 months after Baseline,V6).Results After intervention,the quit rates in intervention group were 12.5％,23.6％ and 20.8％ respectively.Cigarettes per day,times attempting to quit smoking for 24 h and 7 d or more,quit rates were significantly different(P＜0.05).Conclusion The intervention reduces the cigarettes per day,increases the times attempting to quit smoking,advances the stages and increase the quit rate.It is one of the proper choices for smokers and smoking cessation clinic in communities.

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.

AIM:To explore the protective effect of fasudil hydrochloride against cisplatin (CP)-induced renal tubular epithelial cell apoptosis via Akt activation and PTEN inhibition .METHODS:Healthy male Sprague-Dawley ( SD) rats were randomly divided into control group , CP group and CP+fasudil group .All animals were sacrificed 96 h after in-jection of 0.9%saline or CP .Blood samples and kidney tissues were collected to evaluate levels of blood urea nitrogen (BUN), serum creatinine (sCr) and morphological alteration of the kidneys , respectively.The apoptosis of renal tubular epithelium cells was detected by TUNEL.Protein levels of Rho-associated protein kinase 1 (ROCK1), PTEN and Akt were measured by Western blotting and immunohistochemistry .The protein level of p-Akt was analyzed by Western blotting . RESULTS:Compared with control group , the sCr and BUN levels , the expression of ROCK 1 and PTEN and TUNEL-posi-tive cells were increased , while the level of p-Akt was decreased in CP group and CP +fasudil group .The histological structure of the kidneys observed by PAS staining was developed marked structural damage in CP group (P<0.05).Com-pared with CP group, sCr level, the expression of ROCK1 and PTEN and TUNEL-positive cells were decreased, while the level of p-Akt was increased in CP+fasudil group (P<0.05).Very little structural damage was detected in fasudil-treated groups .CONCLUSION:Fasudil hydrochloride has a protective effect on CP-induced renal tubular epithelial cell apoptosis via Akt activation and PTEN inhibition 1.

Objective To evaluate the efficacy and safety of covered metallic stents in the management of proximal esophageal strictures.Methods Retrievable covered self-expanding metallic stents were inserted individually in 43 patients with malignant strictures of the proximal esophagus.Main outcome measures included improvement of dysphagia,sealing of fistula and complications were evaluated.Results A total of 45 stents were placed in 43 patients,with a success rate of 95.6％ (43/45) at the first stent placement attempt.The dysphagia score improved significantly from 3.3 ± 0.6 to 1.8 ± 0.9 (P ＜ 0.001),with 18 having an excellent response and 25 having a good response.Improvement rate of dysphagia was 95.6％ (43/45).Fistula sealing was achieved in 7 of 9 patients (77.8％).Early complications included severe pain (n =5),mediastinal emphysema (n =1),moderate fever (n =9) and collapse of the proximal end of the stent (n =1),while late complications included migration (n =3) and tissue overgrowth (n =6).Conclusion Retrievable covered metallic stent placement is effective and safe for the palliation of obstructive dysphagia and sealing of fistulas in malignant proximal esophageal strictures.