This study aimed at preparing the chemical reference substance of garcinol.A preparative high-speed countercurrent chromatography (HSCCC) was adopted for the isolation of garcinol from the twigs of Garcinia multifiora Champ,G.esculenta Y.H.Li and the fruits of G.xanthochymus Hook.f.ex T.Anders.The crude extracts were separated by HSCCC with two phase solvent systems composed of n-hexane-ethyl acetate-95％ ethanol-water (8∶8∶12∶4,volume ratio) using the upper stationary phase and the lower mobile phase.Under the conditions of a flow rate of 2.0 and 4.0 mL· min-1,and the apparatus rotation at 850 rpm,the column temperature at 25℃ and the detection wavelength at 254 nm,the fraction containing garcinol was purified by the Sephadex LH20 chromatography.The purity test in the extracts was determined by high-performance liquid chromatography.As a result,the contents of garcinol were 45,281 and 4080mg respectively separated from 104.765 g twigs of G.multiflora Champ,105.270 g G.esculenta Y.H.Li and 102.318 g the fruits of G.xanthochymus Hook.f.ex T.with the purity of 98.72％,98.36％ and 98.42％.Compared with the sample pretreatments and separation efficiency of the three plants,it was found that the fat-soluble extracts rapidly and efficiently extracted using n-hexane-ethyl acetate-95％ ethanol-water (5∶5∶5∶5,volume ratio) contained abundant garcinol taking fruits of G.xanthochymus Hook.f.ex T as the raw material with the combination of HSCCC and Sephadex LH-20 chromatography.In conclusion,it was indicated that the combination method is efficient with high operability and large preparation quantity.

Objective To investigate the prevalence rate and related factors of excessive daytime sleepiness (EDS) in the elderly. Methods Epworth Sleep Scales (ESS) was used to make a randomized questionnaire among the retired elderly. Results 1 000 questionnaires were released and totally 768 qualified answers were returned. The mean score of ESS was 4.22?0.10 and the score ≥8 was used as an abmormal value. The prevlence rate of EDS in the elderly group was 10.9%. Regression analysis was done based on ESS score as dependent variable, and age, sex, duration of sleep, difficulty in falling asleep, early wakeup, daytime sleepiness, habitual snoring and loud unstable snoring as independent variables. It was showed that daytime tiredness(r=1.458, P

Background and purpose:It was reported that zinc ifnger protein 139 (ZNF139) was expressed aberrantly in gastric cancer. But the relationship between ZNF139 and multidrug resistance (MDR) of gastric cancer is still not clear. The purpose of this research was to investigate the expressions and signiifcance of ZNF139, MRP-1, MDR1/P-gp, GST-π in human gastric carcinoma cell lines SGC7901 and SGC7901/ADR. Methods: The expressions of ZNF139, MRP-1, MDR1/P-gp, GST-πwere determined with RT-PCR and Western blot in SGC7901 and SGC7901/ADR cell lines. Then siRNA recombinant plasmid of targeting ZNF139 gene was constructed and imported into gastric cancer cell line SGC7901/ADR, and the expressions of MRP-1, MDR1/P-gp, GST-πwere tested simultaneously. Results:The expressions of ZNF139, MRP-1, MDR1/P-gp, GST-πwere higher in SGC7901/ADR than in SGC7901(P<0.05). ZNF139 was inhibited obviously after siRNA-ZNF139 was transfected into SGC7901/ADR, and expression of MRP-1, MDR1/P-gp, GST-πdecreased(P<0.05). Conclusion:ZNF139 may be invovled in multidrug resistance (MDR) of gastric cancer by up-regulating MRP-1, MDR1/P-gp and GST-π.

Objective To analyze basic data and outcomes in Chengdu Stroke Registry.Methods The stroke patients consecutively admitted to Department of Neurology,West China Hospital,Sichuan University since March 1,2002 were prospectively registered.The baseline demographic,risk factors,treatment,and outcome data was recorded with standardized stroke register form by trained specialists.The patients were followed up at seven days,one,three,six months and one year after onset of the stroke for death and disability.Results A total of 3123 consecutive patients were registered between March 1,2002 and August 31,2006,of which 65.5％ came from urban areas and 34.5％ from rural areas.The age was (63.05 ± 17.98) years old and male accounted for 60.3％.Ninety-seven percent (3028/3123) of patients completed CT or MRI scanning during hospitalization.A total of 1804 patients were included between March 2002 and September 2004,of which ischemic stroke accounted for 62.1％ (1120/1804),intracranial hemorrhage 28.4％ (513/1804),subarachnoid hemorrhage 4.0％ (72/1804) and TIA 5.5％ (99/1804).The median NIHSS score on admission was 8(3-15) points in patients with cerebral hemorrhage,and 5(2-10) points in patients with ischemic stroke.Compared with the patients with intracranial hemorrhage,patients with ischemic stroke more frequently had a history of diabetes (OR =2.427,95％ CI 1.811- 3.253,P=0.000),atrial fibrillation (OR=6.121,95％ CI3.535-10.60,P=0.000),coronary heart disease (OR=4.144,95％ CI 2.944-5.832,P =0.000) and TIA (OR=4.342,95％ CI 1.726-10.92,P =0.001 ),and less alcohol consumption ( OR =0.740,95％ CI 0.611-0.896,P =0.002 ).The proportion of in-hospital treatments were thrombolysis 0.9％,anti-platelet therapy 83.0％,mannitol 23.5％,neuroprotective agents (citicoline) 68.1％,and Chinese herbal medicine 89.7％.Case fatality rate was 10.7％ and 13.9％ respectively at 7 days and one month for patients with intracranial hemorrhage,3.0％ and 5.2％ respectively for ischemic stroke.Death or disability was 40.4％,40.3％ and 38.9％ in patients with intracranial hemorrhage and 37.1％,35.0％ and 33.4％ for ischemic stroke at the end of 3,6,12 months respectively.Conclusions Our stroke registry is featured with the largest sample,and the longest period of consecutively registration.It provides an important platform for clinical investigation of stroke.Our study suggested case fatality and disability is lower in this group than in other ethics.Above features should be considered in design of future clinical trials in China.

BACKGROUNDTo investigate the relationship among the CT appearances, the dynamic CT enhancement and the microvessel density (MVD) of peripheral lung cancer.

METHODSThirty-three patients with peripheral lung cancer proved by surgery and pathology underwent enhancement dynamic CT scan before operation, including 14 squamous cell carcinoma and 19 adenocarcinoma. The MVD was measured in resected tumor specimens with immunohistochemical method of LSAB.

RESULTSThe MVD value of adenocarcinoma was significantly higher than that of squamous cell carcinoma (63.4±11.9 versus 50.2±16.3, P < 0.05). The MVD values were higher in junction zone and interstitial areas than those in parenchymal areas, necrotic zones and scar areas of tumors. There were significant relationships among the MVD value and diameter of tumor, lobulation sign, vessel convergence sign, pleural retraction sign and lymph node metastasis (P < 0.05 ). The mean enhanced CT values was (43.4±11.8) HU in adenocarcinomas, and (34.6±10.7) HU in squamous cell carcinomas (P < 0.05). The CT values of both adenocarcinoma and squamous cell carcinoma had positive correlations to their corresponding MVD values (r=0.719, P < 0.01;r=0.819, P < 0.01).

CONCLUSIONSThe CT appearances and the enhanced CT values of peripheral lung carcinomas are closely related to their MVD values, which might be an indicator to identify the histological classification and to predict the malignant degree of tumor.

Biliary Fistula ; diagnosis ; Bronchial Fistula ; diagnosis ; Female ; Humans ; Middle Aged

Objective To investigate the expression of TIP 30 protein in gastric cancer tissues,and effect of TIP30 over-expression on migration and invasion of gastric cancer cell line SGC7901.Methods Immunohistochemistry streptavid-in-peroxidase (SP) methods were used to detect the expression levels of TIP30 in 93 cases of gastric cancer tissues.Previously constructed pcDNA3.1-TIP30 plasmid were transiently transfected into SGC7901 cells.The proliferation of cells were detected by using MTT assay when TIP30 was overexpressed.Transwell assay to determine migration and invasion ability of SGC7901.Western blot was used to examine the changes of concentration of E-cadherin,N-cadherin and MMP9.Results The positive expression rate of TIP30 was 39％ significantly lower in gastric cancer tissues than 92％ in normal gastric mucosa tissues (x2 =32.68,P ＜ 0.05),there was a significant correlation between reduced expression of TIP30 and depth of infiltration,including nodal metastasis,TNM stage (x2 =3.535,7.421,6.754,all P ＜ 0.05);MTT showed that the proliferation of SGC7901 cells in the pcDNA3.1-TIP30 transfected group significantly decreased when TIP30 was overexpressed at respective time of 72,96 hours (t =6.528,7.249,both P ＜ 0.05),Transwell assay showed that overexpression of TIP30 significantly decreased migratory and invasive numbers of SGC7901 cells (t =5.769,P ＜ 0.05;t =7.886,P ＜ 0.05);the expression level of MMP-9 and N-cadherin in TIP30 overexpressing cells group significantly decreased (t =9.811,10.362,both P ＜ 0.05),mean while E-cadherin expression was significantly higher than before (t =6.137,P ＜ 0.05).Conclusion TIP30 protein is low expressed in gastric cancer and the overexpression of TIP30 inhibits the proliferation,migration and invasion of gastric cell line SGC7901.

Objective:To investigate the expression of KLF11 in gastric cancer tissues and cell lines as well as its effect on proliferation and apoptosis of human gastric cancer cells BGC823.Methods:Sixty pairs gastric cancer tissues and corresponding adjacent tissues were collected. The expression of KLF11 mRNA in gastric cancer tissues and their adjacent tissues was detected by RT-PCR. The expression of KLF11 was detected in gastric cancer cells. KLF11 expression was silenced. The proliferation of cells were detected by using MTT assay. Flow cytometry was used to detect the cell cycle and cell apoptosis rate. Western bloting was used to examine the changes of concentration of proteins associated with cell cycle,cell apoptosis and Wnt/β-catenin signaling pathway related proteins. The activity of Caspase3 enzyme was detected by spectrophotometry.Results:The relative expression of KLF11 mRNA in gastric carcinoma tissues was significantly higher than that of the adjacent tissues ( t=11.38, P<0.05). Its expression was related to tumor size, depth of invasion, lymph node metastasis and TNM clinical stage (all P<0.05). The proliferation of BGC823 cells was significantly suppressed after KLF11 silencing ( F=19.56, P<0.05), and the cell cycle was arrested in G 0/G 1 phase [(41.40%±0.98%) vs. (66.53%±1.01%), F=32.69, P<0.05]. Meanwhile, the apoptosis rate was significantly increased by KFL11 silencing [(41.44%±1.59%) vs. (15.42%±0.86%), F=35.35, P<0.05]. The results of Western blotting revealed that the expression of Bax and Cleaved Caspase3 was significantly increased ( F=23.33, 33.63; both P<0.05), wheras that of β-catenin, Bcl-2, CyclinD1and CyclinE was significantly reduced ( F=22.21, 16.24, 26.75, 33.42; all P<0.05). The activity of Caspase3 enzyme was enhanced ( F=16.56, P<0.05). Conclusion:KLF11 was highly-expressed in gastric cancer tissues and cells, KFL11 silencing could inhibit gastric cancer cells proliferation and induce cell apoptosis via Wnt/β-catenin signaling pathway.

Objective:To explore the risk factors of lymph node metastasis (LNM) in early gastric cancer (ECG), and establish a risk-prediction model based on LNM.Method:Four hundred and twenty-seven EGC patients undergoing curative radical gastrectomy were enrolled in this study. The risk factors for LNM of ECG were analyzed with Logistic regression. LNM risk was stratified and risk-predicting model was established. The risk-predicting model was measured by area under ROC curve. According to the same standard, clinical data of 133 patients with EGC who underwent radical surgery were selected for external verification of the model.Results:The frequency of LNM was 13.3% (32/427) in EGC patients. The LNM ratio of intramucosal carcinoma and submucosal carcinoma was 1.3% (3/237), 15.3% (29/190) respectively. Ulcer presence, tumor size >2 cm, undifferentiated tumor, submucosal invasion, neural invasion, and vascular tumor thrombus were significantly associated with LNM in EGC patients ( χ2=3.408, 16.379, 4.808, 29.804, 25.305, 47.120, respectively P<0.05). Multivariate analysis suggested that ulcer presence, tumor size >2 cm, depth of invasion, neural invasion, and vascular tumor thrombus were independent predictors of LNM in EGC patients, ( OR=0.326, 2.924, 11.824, 13.047, 7.756, respectively P<0.05). LNM predicting model is established, P=e^x/(1+e^x),x=-4.792-1.122 ulcer presence+1.073 tumor size+2.470 depth of invasion+2.569 neural invasion+2.048 vascular tumor thrombus,ROC-AUC of risk-predicting model was 0.845, the best cut-off was 0.094, the sensitivity was 72.70%, the specificity was 77.20%. The external verification result revealed the AUC of ROC was 0.840. The four-grid table is constructed by predicting model results and the postoperative pathological examination. The sensitivity and specificity of the model are calculated to be 82.35% and 68.96%, respectively. Conclusions:EGC patients with ulcer presence, tumor size >2 cm, depth of invasion, vascular tumor thrombus, and neural invasion have higher risk of LNM, the risk-predicting model can identify the high probability of LNM .

OBJECTIVETo screen and identify the proteins related with tumor metastasis of gastric cancer in a nude mouse model bearing orthotopic transplanted tumor.

METHODSZinc finger protein 139 (ZNF139)-specific siRNA was synthesized and transfected into gastric cancer cell line SGC7901, which was then screened by G418. ZNF139-siRNA-transfected cells, negative plasmid-transfected cells and untreated SGC7901 cells were orthotopically transplanted separately on the stomach wall of BALB/c nude mice. The primary tumors and metastatic lymph nodes were harvested to separate the proteins by 2-D fluorescence difference gel electrophoresis (2-D DIGE); after gel digestion, the differential proteins were subjected to liquid chromatography-mass spectrometry (LC-MS) for identification and their functions were analyzed. Western blotting was performed to verify the identified proteins.

RESULTSZNF139 expression was effectively inhibited in siRNA-transfected SGC7901 cells. ZNF139-siRNA-transfected cells showed obviously suppressed tumor growth with a lowered lymph node metastasis rate in nude mice compared with untreated cells and the negative control cells (P<0.05). Proteomic study with 2-D DIGE showed that fascin and hnRNPA2/B1 were down-regulated while ANXA1 was up-regulated in the primary tumors, and ANXA5 was down-regulated in the metastatic lymph nodes in ZNF139-siRNA-transfected group. Western blotting confirmed the results of proteomic analysis.

CONCLUSIONZNF139 gene may promote lymph node metastasis of gastric cancer by regulating fascin, hnRNPA2/B1, ANXA1, and ANXA5.

Animals ; Annexins ; metabolism ; Blotting, Western ; Cell Line, Tumor ; Chromatography, Liquid ; Electrophoresis, Gel, Two-Dimensional ; Heterogeneous-Nuclear Ribonucleoprotein Group A-B ; metabolism ; Humans ; Kruppel-Like Transcription Factors ; metabolism ; Lymphatic Metastasis ; Mice ; Mice, Nude ; Neoplasm Proteins ; metabolism ; Neoplasm Transplantation ; Proteomics ; RNA, Small Interfering ; Stomach Neoplasms ; pathology ; Transfection