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 observe the projections from respiratory neurons of medulla oblongata to the motor neuron pool of phrenic nerve in rat. MethodsBy using horseradish peroxidase (HRP) retrograde tracing technique after injecting HRP into phrenic nerve, retrograde labelled neurons were found in C3-C5 segment. Then, HRP was injected into the area where the phrenic motor neurons occupied, retrograde labelled neurons were found in medulla oblongata. ResultsPhrenic motor neurons locate in the C3-C5 segment ipsilaterally, occuping the intermediate portion of anterior horn and appearing typical motor neurons. Retrograde labelled neurons were found bilaterally in medulla oblongata, the neurons were located in nucleus retroambigualis (RNA), ventramedial area to nucleus retrofacialis (NRF). ConclusionThe phrenic motor neurons may receive direct projection of respiratory neuron in medulla oblongata, the projecting neurons are distributed in RNA and NRF area.

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.

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.

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.

Objective:To investigate the relationship between IL-37 and soluble PD-1 in patients with rheumatoid arthritis ( RA) , and investigate the relationship between IL-37 and soluble PD-1.Methods: Peripheral blood was collected from RA patients and control group.The levels of IL-37 and sPD-1 in 30 RA patients (RA standard score ≥6) and 30 healthy controls were measured by enzyme-linked immunosorbent assay ( ELISA).The expression of IL-18, IL-6 and IL-18BP in the peripheral blood of two groups was analyzed and the correlation was analyzed by Pearson correlation analysis .Results:The results of ELISA detection showed that the ex-pression levels of IL-37 and sPD-1 in the peripheral blood of patients with RA and other inflammatory cytokines such as IL -18, IL-18BP and IL-6 were higher than those of healthy controls (P<0.05).IL-37 was positively correlated with cytokines IL-18, IL-18BP and IL-6;sPD-1 was positively correlated with cytokine IL-6.IL-37 and sPD-1 were positively correlated with the severity of patients.Conclusion:IL-37 and sPD-1 are elevated in RA patients and associated with other inflammatory cytokines and disease severi -ty.Both of them may play an important regulatory role in the progress of RA , and provide a new basis for future RA therapy .

BACKGROUND:Studies have indicated that the abnormal expression of tumor necrosis factorreceptor-associated protein 1 (TRAP1) is closely related to the occurrence and development of a variety of tumors. Therefore, targeted inhibition of TRAP1 expression has become an important target for the treatment or intervention of tumor growth. OBJECTIVE:To explore the effect of the TRAP1 gene silencing on the proliferation and apoptosis of human laryngeal squamous cel carcinoma stem cel s. METHODS:CD24-CD44-human laryngeal squamous cel carcinoma stem cel s were isolated by flow cytometry. Interfering RNA (siRNA) sequences for smal molecule TRAP1 gene was designed and transferred into human laryngeal cancer stem cel s by LipofectamineTM 2000. Flow cytometry, MTT assay, cel clone formation assay and TUNEL apoptosis assay were used to evaluate the effect of silencing TRAP1 gene on the proliferation and apoptosis of CD24-CD44+laryngeal cancer stem cel s. RESULTS AND CONCLUSION:Compared with CD24+CD44-cel s, CD24-CD44+cel s upregulated OCT4, SOX2, NANOG and TRAP1 expression levels (P<0.05). However, the expression of TRAP1 protein in human laryngeal squamous cel carcinoma was significantly decreased after RNA interference (P<0.05). The growth rate of TRAP1 gene silenced human laryngeal squamous cel carcinoma was significantly reduced (P<0.05), the cel arrest was in the G0/G1 phase, the number of cel s in the S phase was decreased (P<0.05), and there was no significant change in the M phase. TRAP1 gene silencing significantly inhibited the proliferation of human laryngeal squamous cel carcinoma stem cel s (P<0.05). Compared to the non-transfected cel s, the TRAP1 gene silencing significantly reduced the clone formation ability of transfected human laryngeal squamous cel carcinoma stem cel s (P<0.05), and TRAP1 gene silenced-human laryngeal squamous cel carcinoma stem cel s were more easy to trigger apoptosis by upregulating BAD and BAX expression levels (P<0.05). Overal , our experimental results indicate that the specific interference of TRAP1 gene expression could inhibit the proliferation and promote apoptosis of human laryngeal squamous cel carcinoma stem cel s.