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 investigate in evaluation of various capacity requirement of probationer nurs-ing students by nurses in operation room in order to improve clinical teaching work. Methods A self-de-signed questionnaire was applied, 191 qualified teaching nurses from nine grade three A general hospitals in Beijing were invited to fdl the questionaires. The results underwent subsequent analysis. Results The teach-ers had high expectations for various capacity of the probationer nursing students. Conclusions Clinical teachers should establish feasible teaching scheme according to the existing teaching syllabus in order to in-crease teaching quality, make students gain new insights through reviewing old materials, so that they can mas-ter various technique in operation room in short time.

Objective To explore the blood drug concentration monitoring of sustained-release valproate(DK)in children with epilepsy,focusing on the selection of sampling time and evaluation of the results.Methods Two hundred and seventy-one children taking DK and 155 children taking sodium valproate syrup(VPA Syr)were involved and their serum were taken when achieved steady state to determine the valproic acid level using fluorescence polarization immunoassay.They were divided into 4 groups,which were DK taken once daily group(DK qd group,126 children),DK taken once daily at night and sampled on morning group(DK qn group,26 children),DK taken every 12 h group(DK q12 h group,119 children),VPA Syr q12 h group(155 children).Determine the proportion of the blood drug concentration of each group below,ithin and above the therapeutic range for valproate(50-100 mg/L)were determined.The data were analyzed by t test.Results The Cmin of DK qd group were(73.09?19.91)mg/L,significantly lower from the serum concentration of DK qn and sampled on morning group [(94.94?25.44)mg/L](P0.05).Conclusions DK qn should sampled at night before the night dose.The Cmin of DK q12 h was higher according to the therapeutic range,it's favorable range still needs clinical practice.

Objective To investigate MRI localization the accuracy and correct the deviation for gamma knife treatment.Methods With 25-point-matrix tank,the deviation of MRI localization and its regularities could be identified after the comparison between the coordinates of MRI localization and the ones which have already been verified by CT within the deviation of 0.5 mm.Then the original MRI coordinates will be corrected by the acquired mean deviation and the geometric distortion of images.Afterwards the corrected coordinates will be compared with the standard ones and finally validated by exposure film.Results There are no significant deviations on x-and z-axis after measurement in three hospitals,y-axis,however,bears deviation of (1.94 ±0.45) mm for hospital A,(-2.22 ±0.29) mm for hospital B,(-1.25 ±0.21) mm for hospital C,respectively.Furthermore there also exists geometric distortion of 1％ on y-axis in hospital A.The corrected coordinates on y-axis (Yc) will be calculated from the formula:Yc =(Y-M) + GD (Y0-Y) (Y:the original coordinates on y-axis,M:the mean of deviation on y-axis,GD:the geometric distortion,Y0:the coordinate on y-axis of the central point among the 25-point matrix).Once completed,the corrected coordinates of MRI localization is of no significant difference with the standard coordinates verified by CT.Even the deviation of focal spot on validation film is within 0.5 mm.Conclusions The 25-point-matrix tank in the multi-point measurement of the accuracy and the correction of deviation for gamma knife treatment is feasible to determine whether MRI can be utilized in the localization for head gamma knife treatment.

Objective To explore the clinical value of echocardiography in evaluating the cardiovascular safety of drugs for AIDS patients treated by highly active antiretroviral therapy(HAART).Methods 45 HIV-infected patients accepted three times of echocardiography respectively before the treatment,and after 48 weeks of HAART and 96 weeks of HAART.All kinds of echocardiographic parameters were measured.Results There were statistically significant differences among baseline group pretreatment [V-LVOT:( 115.0 ± 13.8 ) cm/s,EDT:(166.8 ± 26.7) cm/s,E':(26.8±6.7)cm/s],48 weeks [V-LVOT:( 113.0 ± 12.9)cm/s,EDT:(172.9±31.0)cm/s,E':(23.8 ±5.0)cm/s] of HAART and96 weeks[V-LVOT:(107.0±13.3)cm/s,EDT:(140.4 ± 19.3)cm/s,E':(22.7±6.4)cm/s]of HAART for parameters of V-LVOT,EDT and E'( all P ＜ 0.05 ),other echocardiographic parameters showed no statistically significant differences(all P＞0.05).Conclusion Short-term of HAART was provided with cardiovascular safety,the invasive echocardiography could accurately evaluate the cardiac function and structure of AIDS patients with HAART.

Objective To investigate whether Ulinastatin (UTI) would minimize the systemic inflammatory response,lessen cardiac dysfunction and protect neurons against injury in hippocampus CA1area after restoration of spontaneous circulation (ROSC). Methods Animal models of cardiac arrest were established in 24 New Zealand rabbits,and those animals were randomly (random number) divided into control group and UTI treated group after ROSC.Changes in the levels of plasma inflammatory cytokines TNF-α and IL-6 were assayed before cardiac arrest and 4,8,12 and 16 hours after ROSC.Cardiac function including FS,EF and E/A were observed with ultrasonography before cardiac arrest and 4,8,12 and 16hours after ROSC,and viable and apoptotic neurons in hippocampus CA1 area and infiltrations of MPO positive cells in myocardium,cerebrum,liver,kidney and intestine were counted 72 hours after ROSC.The t-test or Mann-Whitney rank sum test was used to verify the specified theoretical distribution functions of the biomarkers tested by Kolmogorov-Smirnov test,POST HOC test was used for the multiple comparisons,and Pearson correlation analysis was used to investigate the correlation between inflammatory cytokines and cardiac function. Results The levels of TNF-α and IL-6 in UTI group were lower than those in control group as those data got 4,8,12 and 16 hours after ROSC (P ＜0.05).EF and E/A in UTI treated group were higher than those in the control group as those data got 4,8,12 hours after ROSC.FS values obtained 4 h and 8 hours after ROSC were higher in UTI group than those in control group ( P ＜ 0.05 ).The Pearson correlation analysis showed that the levels of TNF-α and IL-6 significantly correlated with EF after ROSC.The number of viable neurons in CA1 area of control group was ( 13.22 ± 0.97) which was lower than that in UTI group ( 16.89 ± 1.45 ) ( P =0.003 ),while the number of apoptotic neurons in hippocampus CA1 area was higher in control group than that in UTI group (15.67 ± 1.37) vs.(13.67 ± 1.03 ) (P =0.019).The numbers of MPO positive cells were significantly lower in liver,kidney and intestine in group UTI than those in control group. Conclusions UTI could inhibit the infiltration of MPO positive cells in liver,kidney and intestine,decreasing the levels of TNF-α and IL-6 in plasma,in turn lessening cardiac dysfunction and protecting neurons from injury in hippocampus CA1 area after ROSC of New Zealand rabbits.

Objective To explore the role of flexible fiberoptic bronchoscope(FFB) in the critical patients with severe respiratory diseases of PICU.Methods One hundred and eighty-nine cases of severe respiratory diseases had undergone FFB from Jul 2010 to Jan 2011.One hundred and fifty-seven (83.06％) cases were less than 3 years.FFB examinations were made through nose (226 times) and endotracheal intubation (25 times).Bronchoscopic lavage for bacterial culture was performed by injecting normal saline solution (NSS) via a syringe into the working channel of FFB.Results Of all 189 cases,bacteria were isolated from 80 cases (42.3％) for bacterial culture of bronchoscopic lavage fluid,which included G-bacilli in 51 cases,G + cocci in 21 cases,and fungi in 8 cases.Six cases with tracheobronchial foreign bodies and 31 cases with respiratory tract anatomical abnormalities were found through FFB.Forty cases of severe pneumonia complicated with respiratory failure underwent bronchoscopic lavage,which were effective in 35 cases,and ineffective in 5 cases.In 20 cases of severe pneumonia atelectasis,pulmonary reexpansion were found in 16 cases by bronchoscopic lavage,atomization inhalation,and systemic administration,4 cases improved.Tracheobronchial foreign bodies were removed in all 6 cases by FFB.During FFB,bradycardia was found in 1 cases.There was no FFBrelated death.Conclusion FFB is safe for severe patients with respiratory diseases in PICU.FFB play important role in diagnosis and treatment for children in PICU,including feasible intervention under bronchoscopy.

To investigate the effect of acid and alkali stress on ginsenoside content of Panax ginseng, adventitious roots culture in bioreactors were incubated for 30 d and pH value was adjusted. Ginsenoside content increased by reducing or raising the pH in culture medium, the muxium ginsenoside content was determined on the 5th days after acid treatment and on the 7th days after alkali treatment. The result of histochemical localization of ginsenoside revealed that the red color from light to dark were found in the adventitious root tissue, and ginsenoside mainly located in the pericycle cells where appeared the dark red color.
Ginsenosides ; metabolism ; Hydrogen-Ion Concentration ; Panax ; metabolism ; physiology ; Plant Roots ; metabolism ; Stress, Physiological ; Time Factors

To improve cell suspension culture system of Panax ginseng, the dynamic of cell growth and medium consumption were studied, and the effects of filter on the culture vessel, revolution number, and inoculation density on cell growth and ginsenoside accumulation were also investigated. The maximum cell growth and ginsenoside accumulation was found on the 20th days of suspension culture, therefore, 20 days were confirmed as a suitable culture period for mass production of ginsenoside. Cell growth and ginsenoside content were promoted when the culture vessel had a ventilated filter. Revolution speed during suspension culture affected cell growth, but not ginsenoside content, a peak of ginsenoside productivity was found in the treatment of 120 r x min(-1). Inoculation density also influenced cell growth and ginsenoside accumulation, inoculation density of 6 g was better than other inoculation densities, the ginsenoside content and productivity were up to 12.8 mg x g(-1) DW and 146.6 mg x L(-1), respectively.
Cell Culture Techniques ; methods ; Cell Proliferation ; Culture Media ; chemistry ; Ginsenosides ; metabolism ; Panax ; cytology ; growth & development ; metabolism ; Suspensions

OBJECTIVETo investigate the relationship between Helicobacter species and hepatocellular carcinoma(HCC).

METHODSLiver samples resected during operation from 34 patients with HCC diagnosed by histopathology and 20 without primary liver carcinoma as controls were studied. The two groups of sample were cranked out pathologic slice for in situ hybridization of Helicobacter, Helicobacter pylori and Helicobacter hepaticus. Qualitative and quantitative studies were used to assess the correlation of liver tissue Helicobacter infection with HCC.

RESULTS64.71% (22/34). of the samples of HCC showed positive for Helicobacter specific 16S rRNA-mRNA gene by in situ hybridization, while none was positive in controls (P < 0.01).

CONCLUSIONHelicobacter pylori were found in the liver of patients with HCC.

Carcinoma, Hepatocellular ; microbiology ; Case-Control Studies ; Helicobacter Infections ; complications ; Helicobacter hepaticus ; genetics ; isolation & purification ; Humans ; In Situ Hybridization ; Liver ; microbiology ; Liver Neoplasms ; microbiology