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 explore the efficacy of intraductal chilled saline perfusion (ICSP) to reduce the thermal bile duct injury during the treatment of radiofrequency ablation (RFA) associating with occlusion of hepatic blood supply in rabbits.Methods 16 healthy New Zealand white rabbits were divided into 2 groups.Rabbits of the ICSP group were placed tubes in the common bile duct after laparotomy,and ICSP was performed during the RFA procedure accompanied with hepatic blood occlusion.While for rabbits of the non-ICSP group,hepatic blood occlusion and RFA were performed without ICSP.RFA electrodes were placed about 5 mm away from the hilus hepatis approximately.Contrast-enhanced ultrasonography (CEUS) was administrated to evaluate the sizes of the ablative zones after the procedure.On post-procedure 6 week,ultrasonography was prerformed to evaluate the changes of the biliary structure,and liver specimens of rabbits wcrc obtained for histopathologic observation of main bile ducts.Results Post-procedure CEUS examination showed that there was no significant difference in the size of the ablative zone between the groups (P ＞0.05).On post-procedure 6 week,rabbits of the ICSP group appeared with biliary dilatation more frequently by ultrasonography (P ＜0.05),and a higher degree of the injury of main bile duct by histopathologic observation (P ＜0.05).Conclusions In treatment of RFA accompanied with hepatic blood occlusion,RFA-induced bile duct injury may be decreased significantly with ICSP.

· AlM:To analyze the association between the macular thickness and emmetropic, low myopic, moderate myopic and highly myopic eyes.
·METHODS:The 276 teenagers (276 eyes) between 18~28 years treated in our hospital from January, 2013 to May, 2014 were selected, whose corrective visual acuity was≥1.0 and intraocular preasure was ≤21mmHg and who were willing to participate in this research.Forty-nine emmetropic, 72 low myopic, 104 moderate myopic and 51 highly myopic eyes were measured by optical coherence tomography ( OCT ) to detect the central subfield thickness, bitamporal, superior, lateral and inferior region thickness of inner and outer region, average thickness of retinal macula, foveal thickness and retinal volume.The thickness of different parts of macula lutea was measured and statistically compared among emmetropic, low myopic, moderate myopic and highly myopic eyes.
· RESULTS: The central subfield thickness of emmetropic, low myopic, moderate myopic and highly myopic eyes were (225.38±20.97), (230.97±19.15), (227.01±16.92), (231.91 ±18.97 )μm. The average thickness of retinal macula, of emmetropic, low myopic, moderate myopic and highly myopic eyes were (280.92±12.71), (278.15± 11.90), (270.05±12.07), (267.93±11.08) μm.There were no significant differences of center thickness (F=1.253, P=0.291) and central subfield thickness ( F=1.034, P=0.378) between emmetropic, low myopic, moderate myopic and highly myopic eyes.The macular thickness of inner and outer region in moderate myopic eyes was significantly less than that in emmetropic eyes, and there was significant difference (P<0.05).Comparison between low myopic, moderate myopic and highly myopic eyes was carried out and macular thickness of all regions of inner and outer region in moderate myopic group was significantly increased, and there was significant difference (P<0.05).There was no significant difference when compared with the macular thickness of inner and outer region of the highly myopic group (P>0.05).
· CONCLUSlON: ln low myopic eyes of teenagers, the center macular thickness do not become thinner. However, the macular thickness of inner and outer region is thinner than that of emmetropic eyes.Furthermore, with the increase of the degree of the myopia, the amount of macular thinner gradually decreases from outer region to inner region.

OBJECTIVETo explore the essence of Spleen-Kidney Deficiency in middle-aged patients.

METHODSInvestigation was carried out in 773 cases of 50-69 years old to assay 34 parameters in them, including blood lipid, oxidation and anti-oxidation related substance, sex hormone, liver and renal function, immune function, blood routine, blood pressure and lung vital capacity, etc.

RESULTSSpleen Deficiency Syndrome is closely related with lipid metabolism disorder; Kidney-Qi Deficiency Syndrome is closely related with hypoimmune function; Kidney-Yin Deficiency Syndrome is closely related with lipid metabolism disorder and hypertension, and Kidney Yang Deficiency is closely related with weakness of anti-oxidation capacity, hypoimmunity, the internal environment disorder of sex hormone and aging, manifested as multiple functions abating.

CONCLUSIONSpleen Deficiency, Kidney-Qi Deficiency and Kidney Yang Deficiency are different layers of a gradually developed and aggravated pathological process, but Kidney-Yin Deficiency could not be listed into this layer.

Aged ; Aging ; CD4-CD8 Ratio ; Cholesterol ; blood ; Diagnosis, Differential ; Female ; Humans ; Kidney Diseases ; blood ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Splenic Diseases ; blood ; Superoxide Dismutase ; blood ; Yang Deficiency ; blood ; Yin Deficiency ; blood

Objective To observe the preventive effect of Anduolin(ADL) on radiation-induced lung injury in mice.Methods Totally 180 of Kunming mice were randomly classified into six groups:normal control group ( N ),irradiation control group ( R),irradiation plus low dose ADL group ( L),irradiation plus middle dose ADL group ( M),irradiation plus high dose ADL group ( H),and irradiation plus Dexamethasone group (D).The mice except group N were irradiated with 20 Gy of 6 MV X-rays on whole lung.The mice in group L,M and H were given with ADL 1 d before irradiation and continued for 6 weeks after irradiation.At 2,4 and 6 weeks after irradiation,the general situation and the lung pathological changes of mice were observed.The lung wet weight,collagen contents of the whole lung tissue,hydroxproline concentration,and TGF-β1 expression in the lung were also delected.Results Compared to the group R,the mice breathing rate,hydroxproline concentration,and TGF-β1 expression in the group L were not significantly changed.While in the groups M,H and D,the breathing rate,the generation of hydroxproline and the expression of TGF-β1 were decreased significantly ( F =2.668-161.646,P ＜0.05).In addition,ADL alleviated the pathological changes on radiation-induced lung injury in mice.Conclusions ADL might have the preventive effect on radiation-induced lung injury in mice.

Background:Fenretinide,which is capable of generating reactive oxygen species( ROS ),has emerged as a promising antineoplastic agent based on numerous in vitro and in vivo studies and clinical chemoprevention trials. Preliminary studies showed that fenretinide could induce apoptosis in human hepatocellular carcinoma( HCC)cells in vitro, however,the precise mechanism was not clarified. Aims:To elucidate the effect of ROS on apoptosis of human HCC cells induced by fenretinide and the underlying mechanism. Methods:Human HCC cell line Huh-7 was treated with antioxidant vitamin E,fenretinide or their combination,respectively. ROS in live cells was evaluated by confocal microscopy and flow cytometry;cell viability and apoptosis were assessed by CellTiter-Glo Luminescent Cell Viability Assay Kit and Caspase-Glo3/7 Assay Kit;expression and intracellular localization of nuclear receptor Nur77,as well as expression of stress-induced transcription factor GADD153 were measured by immunofluorescence staining and Western blotting,respectively. Results:Vitamin E pretreatment fully blocked the fenretinide-induced ROS production. In Huh-7 cells pretreated with vitamin E,cell apoptosis induced by fenretinide was significantly reduced(P<0. 05). Furthermore,effect of vitamin E pretreatment was noteworthy on reducing fenretinide-induced GADD153 expression, while no significant impact on fenretinide-induced Nur77 expression and translocation was observed. Conclusions:Elimination of ROS by vitamin E can abrogate the pro-apoptotic effect of fenretinide on Huh-7 cells,which indicates the participation of ROS in fenretinide-induced apoptosis of human HCC cells. Its mechanism might be associated with induction of GADD153 protein expression.

Objective To evaluate the effect of high-level spinal cord injury (SCI) on the myocardial energy metabolism in rats.Methods Sixty healthy male Sprague-Dawley rats,weighing 250-300 g,were randomly divided into 2 groups (n=30 each) using a random number table:sham operation (group S) and SCI group.SCI was induced in anesthetized rats by dropping a 10 g weight onto C7 spinal cord from 5 cm height falling freely inside a vertical hollow glass tube.At 6,12,24,48 and 72 h after SCI,6 rats in each group were chosen and arterial blood samples were taken for measurement of serum creatine kinase (CK) and creatine kinase isoenzyme-MB (CK-MB) activities.The rats were then sacrificed and myocardial specimens were obtained for examination of myocardial ultrastructure and for determination of ATP weight ratio,levels of Na+-K+-ATPase,Ca2+-Mg2+-ATPase,non-esterified fatty acids (NEFA) and lactic acid (LD),and expression of peroxisome proliferator-activated receptor alpha (PPARα) mRNA and protein (using fluorescent quantitative PCR and Western blot).Results Compared with group S,the serum CK and CK-MB activities were significantly increased,the ATP weight ratio,activities of Na+-K+-ATPase and Ca2+-Mg2+-ATPase and levels of NEFA and LD were decreased,and the expression of PPAR-α mRNA and protein was down-regulated in SCI group.No pathological changes of myocardium were found in group S,and the pathological changes of myocardium were obvious in SCI group.Conclusion High-level SCI can lead to decrease in the myocardial energy metabolism in rats,and down-regulated expression of PPARα is involved in the mechanism.

Aim To construct a subtracted cDNA library o f differentially expressed genes in human gastric carcinoma induced by diallyl dis ulfide(DADS). Methods Differentially expressed cDNA species induc ed by DADS in MGC 803 human gastric carcinoma cell line was determined by using suppression subtractive hybridization (SSH). Then these cDNA species were direct ly inserted into T/A cloning vector to set up the subtractive library. Amplification of th e library was carried out with transformation of E.coli by high voltage electrop erforation. One hundred positive bacteria clones were randomly picked and identi fied using PCR method. Results The amplified library contained more than 1,000 positive bacteria clones. Random analysis of 100 clones with PCR m ethod showed that all clones contained 100～600 bp inserts.Conclusions A subtracted cDNA library of differentially expressed genes in MGC 803 hum an gastric carcinoma cell line induced by DADS is constructed successfully with SSH and T/A cloning techniques. The library is efficient and lays solid foundati on for screening and cloning new and specific tumor correlative genes of human g astric carcinoma, and provides a new idea for further exploring the mechanism of DADS effects on carcinoma cells.

Objective To investigate the incidence of gestational diabetes mellitus (GDM) and perinatal outcome in Beijing in 2013. Methods Fifteen hospitals in Beijing were selected by systematic and cluster sampling method. Clinical data of 15 194 pregnant women who attended prenatal care and delivered in these hospitals between June 20 and November 30, 2013 were collected. And 2 987 pregnant women were diagnosed with GDM (GDM group), and 11 999 had no diabetes (non-GDM group). The incidence of GDM, perinatal outcome and complications in mother and infant were compared between the two groups. Two independent samples t-test and Chi-square test were used for statistical analysis. Results The incidence of GDM in Beijing was 19.7%(2 987/15 194). The average age was much higher in GDM group than in non-GDM group [(29.4±4.5) vs (28.0±4.2) years, t=285.705, P<0.05]. The pre-pregnancy weight was much higher in GDM group than in non-GDM group [(59.5±10.2) vs (56.1±8.6) kg, t=352.565, P<0.05]. The incidence of GDM in pre-pregnancy overweight and obese women [29.9%(664/2 230) and 38.8%(250/664)] was much higher than in normal pre-pregnancy weight and low pre-pregnancy weight women [18.0%(1 777/9 890) and 12.9%(273/2 118)] (χ2=296.843, P<0.05). The incidence of GDM in pluripara was higher than in primipara [21.2% (910/4 298) vs 19.4%(2 077/10 688),χ2=5.813, P<0.05]. The incidence of GDM in gradeⅢhospitals was higher than in grade Ⅱ hospitals [21.1% (1 654/7 849) vs 18.7% (1 333/7 173), χ2=13.440, P<0.05]; and the incidence in urban areas was higher than in rural areas [21.3%(2 028/9 521) vs 17.1%(896/5 249),χ2=39.137, P<0.05]. The rate of cesarean section was 47.1%(1 407/2 987) in GDM group, significantly higher than in non-GDM group [39.8% (4 782/11 999)] (χ2=72.204, P<0.05). The incidences of preterm labor [7.3%(217/2 987)], hypertensive disorders [6.3%(185/2 987)], large-for-gestational-age infants [9.2%(275/2 987)], macrosomia [9.5%(283/2 987)] and neonatal ward admission [8.6%(258/2 987)] were all significantly higher in GDM group than in non-GDM group [5.7%(686/11 999), 3.9%(454/11 999), 5.8%(694/11 999), 7.2%(861/11 999), and 6.5%(778/11 999), respectively] (χ2=10.117, 34.371, 79.378, 20.346 and 17.236, respectively, all P<0.05). Conclusions The incidence of GDM is still high in Beijing, and advanced maternal age and pre-pregnancy overweight or obesity are high risk factors for GDM. The rate of preterm labor, hypertensive disorders, and macrosomia in GDM group is higher than in normal fasting glucose group. Systematic obstetric care for GDM should be intensified in Beijing.

ObjectiveTo investigate the influence of pre-pregnancy body mass index (BMI) and gestational weight gain (GWG) on lipid variations of first and third trimester.MethodsA retrospective analysis was conducted on 2 593 singleton pregnant women without diabetes from 15 hospitals in Beijing from June 20 to November 30 in 2013. Serum lipid levels were determined in all of these women at the first trimester (<14 weeks) and the third trimester (≥28 weeks). Demographic data and medical information were collected individually through questionnaires. The recruited gravidas were divided into normal pre-pregnant BMI group (BMI<25) and pre-pregnant overweight and obese group (BMI≥25) according to recommendations of the Institute of Medicine (IOM). Based on the same recommendations on normal GWG, the pregnant women in the two groups were further assigned to two subgroups including normal GWG and over GWG group. The serum lipid levels in both first and third trimester and the increases from first to third trimester were compared between the normal and abnormal BMI groups with two independent samplet-test.Results(1) The levels of total cholesterol (TC), triglyceride (TG), high density lipoprotein-cholesterol (HDL-C) and low density lipoprotein-cholesterol (LDL-C) in the third trimester were all significantly higher than those in the first trimester in the normal BMI group [(6.47±1.17) vs (4.63±0.80), (3.14±1.19) vs (1.22±0.70), (1.94±0.44) vs (1.85±0.49) and (3.56±1.00) vs (2.29±0.67) mmol/L,t=-61.250,-65.909,-6.448 and-49.952, allP<0.01]. And the same results were shown in the abnormal BMI group [(6.20±1.14) vs (4.75±0.88), (3.31±1.20) vs (1.44±0.70), (1.86±0.44) vs (1.65±0.37) and (3.30±0.89) vs (2.51±0.82) mmol/L, t=-18.756,-25.165,-6.905 and-12.191,allP<0.01]. Significant differences were identified between the normal and abnormal pre-pregnant BMI groups both in first and third trimester (allP<0.05). (2) The increases of TC from early to late pregnancy in the abnormal BMI group was lower than that in the normal BMI group [(1.45±1.01) vs (1.84±1.02) mmol/L,t=6.664,P<0.01], but the increase of HDL-C was higher [(0.21±0.39) vs (0.09±0.51) mmol/L,t=-4.301,P<0.01]. (3) In the normal BMI group, the increase of TC and HDL-C between the third and the first trimester was lower in the over GWG subgroups than in the normal GWG subgroups [(1.77±0.96) vs (1.89±1.05) mmo/L,t=-2.802,P=0.005; (0.06±0.41) vs (0.12±0.58) mmo/L,t=-2.588,P=0.010], but the increase of TG was higher [(2.00±1.07) vs (1.86± 1.18) mmo/L,t=3.015,P=0.003]. While in the abnormal BMI group, smaller increase of HDL-C was found in the over GWG subgroup than in the normal GWG subgroup [(0.17±0.35) vs (0.29±0.44) mmo/L,t=-2.664, P=0.008].ConclusionsThe lipid levels during pregnancy are significantly higher in late pregnancy than in early pregnancy. Pre-pregnancy BMI has much more effect on lipid variations during pregnancy than GWG, which primarily affect the lipid levels of women with normal pre-pregnancy BMI.