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

Aged ; Deglutition Disorders ; etiology ; Humans ; Male ; Zenker Diverticulum ; complications ; pathology

Objective: To explore the expression and gene amplification status of human epidermal growth factor receptor-2 (HER2) in the different stages of invasive urothelial bladder carcinoma. Methods:Tumor tissues from 49 patients with different stages of invasive urothelial bladder carcinoma were tested by immunohistochemical staining for HER2 and HER2 gene fluorescence in situ hybridization. Results:The number of male patients was higher than that of females. The positive rate of HER2 protein expression was higher in the patients with the higher stage of invasive urothelial bladder carcinoma. However, no gene amplification was observed in all patients. Twelve patients had ployploid chromosome 17. More ployploids were observed in the patients with the higher stage of inva-sive urothelial bladder carcinoma. Conclusion:The increase in the protein expression of HER2 in the invasive urothelial bladder carci-noma patients was not caused by gene amplification. Other transcriptional and post-transcriptional mechanisms were probably involved in the regulation of the HER2 protein.

Objective To investigate the relationship between IgG antibody titer in pregnant women with maternal-fetal ABO blood incompatibility and hemolytic disease of fetuses and newborns.Methods From January 31 2009 to January 31 2010,1269 singleton pregnant women who were suspected to have maternal fetal ABO blood incompatibility in Department of Obstetrics and Gynecology,Southwest Hospital,Third Military University were collected.Anti-A or anti-B IgG titers of them were detected at 28-30 gestational age,and umbilical cord blood were taken when delivery and hemolytic disease of the newborn serological test were done to diagnose hemolytic disease of the newborn (HDN).The relationship between the titers and incidence of fetal or neonatal hemolytic disease was retrospectively analyzed by Kendall tau rank correlation.Results No IgG of anti-A or anti-B in serum were found in 58.4％ (741/1269) pregnant women,while the antibody titer of 5.1％ (65/1269) pregnant women were more than or equal to 1 ∶ 128.When they were tested again at 36 gestational week,the titer of 17 cases increased twice but lower than 1 ∶ 512.No signs of intrauterine hemolysis,such as edema,ascites and pleural effusion,were found.Three hundred and eighty neonates (29.9％,380/1269) were diagnosed as HDN.Among which,12 cases (3.2％,12/380) showed mild anemia and (or) jaundice within 24 hours after delivery.There was positive correlation between incidence of neonatal hemolysis and antibody titer(Tb=-0.293,P＜0.01).The incidence of HDN increased from 85.4％ (35/41) in women with antibody titer of 1 ∶ 128 to 5/5 inwomen with antibody titer at 1 ∶ 512 (x2=108.906,P＜0.01).Among 380 HDN neonates,322 cases were transferred to neonatal intensive care unit for phototherapy based comprehensive therapy,and two underwent exchange transfusion.All patients were cured.Conclusions The intrauterine hemolysis incidence of patients with suspected maternal-fetal ABO blood incompatibility is very low,and no special care is required during pregnancy.Anti-A or anti-B tests during pregnancy is helpful in early diagnosis and management of HDN.

Ectodermal Dysplasia ; classification ; diagnosis ; Humans ; Infant ; Male ; Patient Care

Objective To survey the CT radiation dose to cancer patients,and to compare it with diagnostic reference level (DRL) provided by newly issued national standard.Methods Computed tomography dose index weighted (CTDIw) of 5 CT equipments was measured by the standard phantom with typical scanning protocol and radiation dose parameters including average computed tomography dose index volume (CTDIvol),dose length product (DLP),75％ DLP with 900 adult and 300 pediatric cancer patients were collected.According to age stage,effective dose was estimated.Results The measured CTDIw for head scanning was on the same level with DRL,while lumbar vertebra and abdominal scanning were lower than DRL.To adults,DLP of cerebrovascular enhanced scanning and effective dose of abdominal enhanced scanning were the highest.To pediatric patients,there was no significant difference in DLP among different ages.Conclusion CTDI only reflects the dose contribution from a slice,but do not reflect the cumulative dose.For the patients need multiphase scanning,because they may be exposed to high dose,national standard should provide DLP value as the diagnostic reference level.Using individual scanning protocol is an effective method to reduce radiation dose of pediatric patient.

Objective To study the differential expressions of labour-associated genes,including interleukin-8 (IL-8),oxytocin receptor (OTR) and prostaglandin H synthase type-2 (PGHS-2)) between preterm labour and term labour myometrium and their clinical significance.Methods Seventeen cases of preterm labour (PTL) and 31 cases of preterm no labour (PTNL) of pregnant women were selected as study group,while 6 cases of term labour (TL) and 6 cases term no labour (TNL) of pregnant women were chosen as control.Lower myometrial tissue biopsies were collected to detect the expressions of IL-8,OTR and PGHS-2 by real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) as well as Western blot analysis.The expression data were then statistically analyzed in combination with clinical data.Results In the lower myometrial tissues,the mRNA levels of IL-8 and PGHS-2 were significantly lower in PTL group than in TL samples (P0.05),but the PGHS-2 mRNA level was significantly higher in TL group than in TNL group (P0.05).Correlation analysis suggested that the mRNA level of IL-8 was significantly correlated with gestational age in PTNL group (r=0.294,P=0.042).Multiple linear regression analysis showed a mutual control effect of the expressions of IL-8,OTR and PGHS-2 in preterm but not in term labour group.Conclusion Our results show that there are significant differences of the expression of labour-associated genes in lower myometrum between preterm labour and term labour,which might becorrelated with the progression of preterm labour.

Objective To study the model of separation and culture and the growth characteristics and osteogenic capability of bone marrow stromal stem cells (BMSCs). Methods BMSCs were isolated from adult rats using gradient centrifugation and anchoring culture. The passage cells were induced to differentiate into osteoblasts by means of an differentiation induction medium containing dexamethasone,?-glycerophophate and Vitamin C. Alkaline phosphatase (ALP) activity and the ability to form mineralized nodules were examined. Results Colonies of stromal stem cells were formed in the primary culture and contacted one another at the 14th day. In the passage culture, cells became bigger than those in the primary culture and could be subcultured one generation in 5～7 days. After the culture in the differentiation induction medium, the ALP activity was enhanced significantly and the mineralized nodules were formed. Conclusion BMSCs can be obtained easily and have strong proliferation ability and osteogenic capacity. This study suggests that BMSCs can become seeding-cells in bone tissue engineering.

Objective To investigate the relationship between the promoter of IL-12B gene polymorphism and the susceptibility and clinical features of chronic gastritis and duodenal ulcer with or without Helicobacter pylori(Hp) infection in children and adolescent.Methods Mucosal biopsies were obtained from 132 children and adolescent (patient group),including 100 children with chronic gastritis and 32 children with duodenal ulcer,undergoing an upper gastrointestinal endoscopy for dyspeptic symptoms.Biopsy specimens were stained with hematoxilin and eosin (HE),and gastritis was graded according to the Sydney system.Serology,urease test and histology were taken to assess Hp status.Genomic DNA was obtained from peripheral blood or gastric biopsies of patients and 102 healthy children as normal control group.The promoter of IL-12B +1188A/G gene polymorphism was genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and sequencing.The genotype distributions and allele frequencies were compared between the study group and the normal control group,and the association of genotypes with clinicopathological features was studied.IL-12B mRNA level expressions in gastric mucosa were confirmed by reverse transcription PCR biopsy-based tests.Results The genotype distributions and allele frequencies of IL-12B +1188A/G gene polymorphisms were similar in gastric upper gastrointestinal diseases and healthy subjects.The IL-12B +1188A/G gene polymorphisms were not associated with Hp status.IL-12B+1188A/G gene polymorphisms did not affect IL-12B mRNA level expressions and were not associated with the degree of antrum chronic inflammation.Conclusions These data suggest that IL-12B+1188A/G gene polymorphisms are not associated with susceptibility to chronic gastritis and duodenal ulcer in children and adolescent.

OBJECTIVETo evaluate the clinical efficacy and safety of phosphodiesterase 5 (PDE-5) inhibitors for erectile dysfunction (ED) in patients with diabetes mellitus and provide some evidence for the clinical treatment of the disease.

METHODSWe searched MedMed, EMbase, Cochrane Library, CNKI, Wan Fang Data, VIP and ZADL for randomized controlled trials on PDE-5 inhibitors for ED in diabetic men and evaluated the methodology of the included trials with the Jadad scale. We used the erectile function domain in the IIEF (IIEF-EF), IIEF questions (IIEF-Q) 3 and 4, SEP-2 and -3, and Global Assessment Questions (GAQ) as the main evaluation indexes and employed the Review Manager 5. 1. 0 software for meta analysis.

RESULTSA total of 13 studies were included, which were all high quality trials with Jadad score > 3. The IIEF-EF scores in 10 of the included studies were subjected to meta analysis using the random-effect model (REM), with a weighted mean difference (WMD) of 5.64 (95% CI 4.41 - 6.83, P < 0.001). The fixed-effect model (FEM) analysis of the IIEF-Q scores in 6 of the studies showed the WMD to be 0.96 (95% CI 0.83 -1.08, P < 0.001) for IIEF-Q3 and 1.11 (95% CI 0.98 - 1.25, P < 0.001) for IIEF-Q4. FEM analysis of the SEP-2 scores showed WMD = 17.67 (95% CI 12. 38 - 22. 97, P < 0.001) in 2 of the studies, and that of the SEP-3 scores WMD = 23.64 (95% CI 17. 49 - 29.79, P < 0.001) in 5 of the studies. The GAQ scores in 11 of the studies were subjected to REM analysis, with OR = 6. 20 and 95% CI 3.65 - 10.52 (P < 0.001). REM analysis was performed on the adverse reactions in 11 of the studies, with OR = 7.43 and 95% CI 4.11 - 13.44 (P < 0.001).

CONCLUSIONPDE-5 inhibitors can effectively and safely improve erectile function in patients with diabetes mellitus.

Diabetes Mellitus ; Erectile Dysfunction ; drug therapy ; Gangliosides ; Humans ; Male ; Penile Erection ; Phosphodiesterase 5 Inhibitors ; therapeutic use