OBJECTIVETo discuss the mechanism of gambogenic acid (GNA) in inducing the apoptosis of melanoma B16 cells.

METHODThe inhibitory effect of GNA on the proliferation of B16 cells was measured by the methyl thiazolyl tetrazolium (MTT) assay. The effect of GNA on B16 cells was detected by the Hoechst 33258 staining. The transmission electron microscopy was used to observe the ultra-structure changes of B16 cells. The changes in PI3K, p-PI3K, Akt, p-Akt, p-mTOR, PTEN proteins were detected by the Western blotting to discuss the molecular mechanism of GNA in inducing the apoptosis of B16 cells.

RESULTGNA showed a significant inhibitory effect in the growth and proliferation of melanoma B16 cells. The cell viability remarkably decreased with the increase of GNA concentration and the extension of the action time. The results of the Hoechst 33258 staining showed that cells processed with GNA demonstrated apparent apoptotic characteristics. Under the transmission electron microscope, B16 cells, after being treated with GNA, showed obvious morphological changes of apoptosis. The Western blot showed a time-dependent reduction in the p-PI3K and p-Akt protein expressions, with no change in p-PI3K and p-Akt protein expression quantities. The p-mTOR protein expression decreased with the extension of time, where as the PTEN protein expression showed a time-dependent increase.

CONCLUSIONGNA could inhibit the proliferation of melanoma B16 cells and induce their apoptosis within certain time and concentration ranges. Its mechanism in inducing the cell apoptosis may be related to PI3K/Akt/mTOR signaling pathways.

Animals ; Apoptosis ; drug effects ; Blotting, Western ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cell Survival ; drug effects ; Dose-Response Relationship, Drug ; Melanoma ; metabolism ; pathology ; ultrastructure ; Mice ; Microscopy, Electron, Transmission ; Microscopy, Fluorescence ; PTEN Phosphohydrolase ; metabolism ; Phosphatidylinositol 3-Kinases ; metabolism ; Proto-Oncogene Proteins c-akt ; metabolism ; Signal Transduction ; drug effects ; TOR Serine-Threonine Kinases ; metabolism ; Terpenes ; pharmacology ; Xanthenes ; Xanthones ; pharmacology

Objective To retrospectively study the methods and effects of interventional therapy in primary hepatic carcinoma complicated with arteriovenous fistula. Methods Interventional chemotherapy and embolization was applied for treatment of 46 cases with primary hepatic eereinoma complicated with arteriovenous fistula. Postoperative clinical changes were observed. Results All 46 cases were successfully treated. No severe complication was found. Total effective rate was 95.65%. 1-year survival rate was 75.09%. Conclusion Interventional treatment is safe and effective in treating primary hepatocellular carcinoma complicated with hepatic arteriovenous fistula.

The exam under open and close condition is a new mode which is different from traditional close-exam and entire open-exam.It indicates that new exam mode not only conquers limitation of simplex close-exam,but also boosts forwardly study of students and promotes the didactical reform completely.

OBJECTIVETo compare the differences in pain reaction, hemodynamics and clinical efficacy between extracorporeal shock wave lithotripsy (ESWL) after injection with flurphen mixture (mixture of droperidol and fentanyl citrate) at Shenshu (BL 23) and simple ESWL in the patients.

METHODSSixty-four cases of urinary calculi with ESWL were randomized into an observation group and a control group, 32 cases in each one. In the observation group, 15 to 20 min before ESWL, flurphen mixture (droperidol injection 1.25 mg and fentanyl citrate injection 0.05 mg were diluted to 6 mL with 0.9% sodium chloride solution 4.5 mL) was injected at bilateral Shenshu (BL 23). In the control group, no any adjuvant therapy and medication were used before ESWL. The changes in blood pressure and heart rate, visual analogue scale (VAS) score, lithotripsy frequency till calculi complete removal and the rate of calculi complete removal after the first lithotripsy were observed in the two groups.

RESULTSIn the control group, blood pressure and heart rate were higher during lithotripsy than those before lithotripsy (both P<0.05). In the observation group, the differences in blood pressure and heart rate were not significant statistically as compared with those before lithotripsy (both P>0.05). The blood pressure and heart rate during lithotripsy in the observation group were apparently lower than those in the control group (both P<0.05). VAS scores during lithotripsy in the observation group were lower apparently than those in the control group (both P<0.05). The lithotripsy frequency in the observation group was less than that in the control group. The rate of calculi complete removal in 1 week after the first lithotripsy in the observation group was higher than that in the control group [75.0% (24/32) vs 50.0% (16/32), P<0.05].

CONCLUSIONThe flurphen mixture at Shenshu (BL 23) significantly alleviates pain reaction in patients undergoing ESWL, avoids the fluctuation of hemodynamics and improves the clinical effect of lithotripsy.

Acupuncture Points ; Adolescent ; Adult ; Aged ; Analgesics ; administration & dosage ; Female ; Hemodynamics ; Humans ; Kidney Calculi ; therapy ; Lithotripsy ; adverse effects ; Male ; Middle Aged ; Pain ; drug therapy ; etiology ; Young Adult

Humans ; Male ; Middle Aged ; Neurofibroma ; pathology ; Pharyngeal Neoplasms ; pathology

Objective To construct pcDNA3-HIF-1? eukaryotic expression vector and to investigate its function in breast cancer MCF-7 cells.Methods RT-PCR was applied to amplify human HIF-1? cDNA from MCF-7 cells with a pair of sequence specific primers carrying a restriction enzyme site XbaⅠ or HindⅢ on each 5′end.HIF-1?(cDNA) was inserted into pMD18-T after sequencing,and then inserted into pcDNA3 vector.The recombinant vector was transfected into MCF-7 cells to observe its expression and function by Western blot and dual-luciferase reporter gene assay.Realtime-PCR was performed to detect the inducible expression of C-MET mRNA by HIF-1?,the anti-apoptotic effect of HIF-1 under hypoxia was analysed by detecting the Caspase3/7 activity.Results The sequence of pcDNA3-HIF-1? is identical with the gene bank.It enhances the expression of HRE reporter gene and C-MET mRNA,but decreases the Caspase3/7 activity in MCF-7 cells under hypoxia.Conclusion The pcDNA3-HIF-1? eukaryotic expression vector was successfully constructed,it not only has strong DNA binding and inducing activity,but also has anti-apoptotic effect in hypoxia MCF-7 cells.

Objective To construct pGL3-EPO-HRE reporter gene vector and to detect its function.Methods To synthesize two single-strand DNA fragments which had overlap sequence at the 3'end,then mixed them to construct the double-strand DNA fragment which include three copies of(57 bp) human erythropoietin(EPO) HRE.There was MluⅠor BglⅡ digestive site at the end of this DNA fragment.After inserting the(87 bp) fragment into pMD18-T for sequencing,the correct sequence was inserted into pGL3-promoter to construct pGL3-EPO-HRE.The recombinant vector was co-transfected into MCF-7 cells with pcDNA3-HIF-1?or pcDNA3 respectively,(12 h) later cells were treated with(100 ?mol/L) cobalt chloride(CoCl_(2)) for(24 h), then the expression of pGL3-EPO-HRE was observed by dual-luciferase reporter gene assay method.Results The sequence of pGL3-EPO-HRE was identical with human EPO HRE sequence.pGL3-EPO-HRE luciferase activity increased about 10 fold(P

The pathogenic effects of crops from Keshan disease area were studied by rat-feeding. As double controls, another two groups were fed with crops from non-endemic area and synthetic diet. Those from disease area caused necrosis of heart muscle vascular changes of myocardial mitochondria and fatty degeneration of liver. Compared with the controls, the free radical and lipid peroxide content in heart and liver were significantly higher, while the ATP and AN contents in these organs were apparently lower. The multielement contents and its constituent patterns in the feeds, whole blood, heart, liver, kidney and brown adipose tissue of rats in experimetal group were apparently different from those in the control groups. The statistic analysis of multiple factors showed that eight elements Se, Mo, Cu, Mn, Cr, Fe, Zn and Pr might be the perilous factors, that is, under the condition of low selenium, the constituent pattern of selenium and the eight elements might be the complicated factors of endemic crops.

Objective To investigate ectopic pregnancy from embryo transfer (ET)of in-vitro fertilization (IVF) cycle and intracytoplasmic sperm iujection (ICSI) cycle and frozen-thawed (FET) cycle.Methods From Jan.2005 to Dec.2010,a total of 9037 IVF-ET or ICSI-ET cycles and 4034 FET cycles were performed in our reproductive medicine center,Affiliated First Hospital of Zhengzhou University.The incidence of ectopic pregnancy rate was studied in fresh cycles IVF-ET (5998) and ICSI-ET (3039) cycles,and natural FET (2198) and hormone replacement (E-P) FET (1836) cycles.ResultsOf 4034 FET cycles,1090 clinical pregnancies and 26 ectopic pregnancies were observed,the incidence of ectopic pregnancy was 2.38％ (26/1090).Of 9037 fresh cycles,3602 cycles were clinical pregnancy,and 133 cycles were ectopic pregnancy,and the incidence of ectopic pregnancy was 3.69％ (133/3602).The ectopic pregnancy rate in FET cycles was lower than in fresh cycles significantly (P ＜0.05).Of 3039 fresh ICSI-ET cycles,the incidence of ectopic pregnancy was 2.62％ ( 34/1298 ) in 1298 clinical pregnancies.Of 5998 IVF-ET cycles,2304 clinical pregnancies were observed,the incidence of ectopic pregnancy was 4.30％ (99/2304).Ectopic pregnancy rate in the fresh ICSI-ET cycles was lower than that of IVF-ET group significantly (P ＜ 0.01 ).The ectopic pregnancy rate in the natural FET cycles was 1.46％ (8/547),which was significantly lower than 3.31％ ( 18/543 ) in E-P group ( P ＜ 0.05 ).ConclusionsThe incidence of ectopic pregnancy of FET cycles was significantly lower than that of fresh embryo transfer cycles.The application of exogenous sex hormones in assisted reproductive cycles might increase occurrence of ectopic pregnancy.

Objective To study the ways of auto swivel reposition under Barium meal for treating stomach volvulus and its clincial value.Methods The patients with stomach volvulus confirmed by gastroenterography underwent reposition according to the volvulus and the effect was observed.The cause was examined.Results 100 cases underwent treatment of auto swivel reposition under Barium meal,among whom 98 cases were successfully repositioned and 51 cases were known for the cause.Conclusion Auto swivel reposition under Barium meal for treating stomach volvulus and its clincial value is simple and non-invasive and acceptable,being of great value.