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

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.

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.

Objective To evaluate the clinical value of 64-slice spiral CT,MRI + magnetic resonance cholangiopancreatography (MRCP) on suspected biliary obstruction.Methods Eighty-five patients with suspected biliary obstruction were examined by 64-slice spiral CT,MRI + MRCP,compared with the pathological results.The accuracy rate of CT,MRI + MRCP,CT + MRI + MRCP in evaluating the sites and nature of obstruction were calculated.Results The accuracy rate of evaluating the site of obstruction in CT was 96.5％ (82/85),MRI + MRCP was 97.6％ (83/85),difference was not statistically significant (P ＞ 0.05).The accuracy rate of evaluating the nature of obstruction in CT was 81.2％ (69/85),MRI + MRCP was 87.1％ (74/85),CT +MRI + MRCP was 94.1％ (80/85),there was no statistically significant difference between MRI + MRCP and CT with CT + MRI + MRCP (P ＞ 0.05),difference was statistically significant between CT and CT + MRI + MRCP (P ＜ 0.05).Conclusion A comprehensive analysis of CT,MRI + MRCP can increase the accuracy of diagnosis biliary obstruction.

As children are extremely sensitive to the bad taste of medicine,they have poor compliance with the bitter medicine.It is of great importance to develop the approaches of taste masking for the research of paediatric drug formulations.Besides,taste masking technology is one of the main barrier to develop children pharmaceutic preparation.This article provides an overview of the advance in taste masking technology of oral paediatric medicine in recent years,and introduces five types of taste masking technology in terms of drug,preparation and the bitter taste transduction,including principle and characteristics of these approaches,as well as their application in formulations,so as to provide some references for the development of paediatric medicine.

Objective To explore the effect of endotracheal intubation therapy on acute laryngotracheobronchitis and acute laryngitis in children.Methods Twelve of 45 children with acute laryngotracheobronchitis or acute laryngitis were intubated and mechanically ventilated in pediatric intensive care unit (PICU) during Oct. 2000 to Oct. 2006.Ten cases were laryngotracheobronchitis and 2 were acute laryngitis.The endotracheal tube chosen was at least 0.5 to 1.0 size smaller than normally recommended for age.The need of intubation was assessed based on severity of signs of airway obstruction Ⅲ without improvement by the conventional treatment with humidified and combined with oral or intravenous interventions,respiratory failure,or airway obstruction Ⅳ.Results Eleven of 12 children were intubated successfully,except 1 case after resuscitation.All the patients were ventilated after intubation.The mean duration of intubation was 3.4 days (24 hours -9 days).Ten cases were successfully extubated and 2 cases failed of extubation,of which 1 case got tracheostomy.All children were routinely supported by nasal continuous positive airway pressure(NCPAP) after extubation with a mean of 1.5 days (2 hours-4 days).The mean days of hospitalization were 11.2 days (4-22 days).Conclusion Intubation is an alternative for the management of severe acute laryngotracheobronchitis and acute laryngitis in children.In most cases,tracheotomy can be substituted by intubation.

Objective:The aim of this study was to investigate the role of cyclic adenosine monophosphate (cAMP) signaling on the differentiation of stem cells from the apical papilla (SCAPs).Methods:SCAPs were cultured by enzyme digestion method.After adding activator (Forskolin) and inhibitor (H-89) into the culture of SCAPs in mineralization-inducing medium,calcium deposition was measured by alizarin red staining.QPCR was performed to measure the mRNA expression of mineralization-related genes.Results:When the cAMP signaling was activated,calcium deposition and the mRNA expression of ALP,OCN,OSX and RUNX2 increased sharply.After inhibiting the signaling,calcium deposition and the mRNA expression of ALP,OCN,OSX and RUNX2 decreased.Conclusion:The cAMP signaling may promote the odontogenic and osteogenic differentiation of SCAPs.

Objective To study the effect of low-dose dexamethasone after acute and subacute cerebral infarct.Methods One hundred and forty patients with acute ischemic stroke were randomly divided into acute ischemic stroke group (acute stage group),subacute ischemic stroke group (subacute stage group),placebo and control groups.Subjects in acute stage groups received conventional therapy and 1 mL (5 mg) dexamethasone injection from 1 day to 3 day after admission and in subacute stage group,received the same treatment as acute stage group from 4 day to 6 day after admission.In control and placebo groups,subjects received conventional therapy and conventional therapy + 1 mL normal saline for injection respectively.One week after treatments,complete blood count and erythrocyte sedimentation rate were tested.One month and three month after treatments,neurological function were evaluated by Barthel Index and modified Rankin Scale (mRS).Results The valuate of erythrocyte sedimentation rate in acute stage group were significantly different from it of placebo and control groups (P ＜ 0.05).Moreover,neurological function of subjects in acute stage group was significantly improved than that in placebo and control groups in by Barthel Index and mRS (P ＜ 0.05).However that in subacute stage group was not different from that placebo and control groups (P ＞ 0.05).Conclusion Low-dose dexamethasone plays a neuroprotection role after acutc cerebral ischemia.

ObjectiveTo investigate the expressions ofheparinase,matrix metalloproteinase 2 (MMP2) and tissue inhibitor of metalloproteinase 2(TIMP2) in malignant melanoma lesions and their significance.MethodsSkin specimens were obtained from the lesions of 30 patients with malignant melanoma,30 patients with melanocytic nevus and the normal skin of 15 healthy controls.Immunohistochemical SP method was used to detect the protein expression of heparinase,MMP2 and TIMP2.ResultsThe malignant melanoma tissue specimens significantly differed from the melanocytic nevus and control tissue specimens in the expression rate of heparinase (63.33％ vs.6.67％ and 0.00,x2 =21.172,27.805,both P ＜ 0.01 ),MMP2 (70.00％ vs.13.33％ and 0.00,x2 =19.817,19.866,both P＜ 0.01) and TIMP2(60.00％ vs.6.67％ and 0.00,x2 =19.200,15.000,both P ＜ 0.01 ).ConclusionThe expression of heparinase,MMP2 and TIMP2 is significantly higher in malignant melanoma lesions than in melanocytic nevus lesions and normal skin tissue.

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.