Objective To observe the effects of ursolic acid (UA) on the activation of nicotinamide adenine dinucleotide phosphate oxidase (NOX) and the downstream signaling pathways in platelet derived growth factor (PDGF) activated rat hepatic stellate cell (HSC-T6).Methods Rat HSC-T6 cells were divided into blank control group (no treatment),UA control group (50 μmol/L UA),PDGF group (10 μg/L PDGF),UA intervention group (50 μmol/L UA + 10 μg/L PDGF),diphenyleneiodonium intervention(DPI) group (20 μmol/L DPI+ 10 μg/L PDGF),SB203580 (p38 mitogen-activated protein kirase(p38MAPK) inhibitor) intervention group (10 μmol/L SB203580 + 10 μg/LPDGF),LY294002 (phosphatidylinositop 3 kinase(PI3K) inhibitor) intervention group (10 μmol/L LY294002 + 10 μg/L PDGF) and rosup positive control group (5 μg/mL rosup).Except rosup positive control group,the expression of type Ⅰ collagen at mRNA level of each group was detected by fluorescence quantitavepolymerase chain reaction (RT-PCR).The expression of membrane protein p47phox (except rosup positive control group),PI3K(except rosup positive control group and SB203580 intervention group),p-protein kinase B (p-AKT,except rosup positive control group and SB203580 intervention group) and phosphorylated p38 mitogen-activated protein kinase (p-p38MAPK,except rosup positive control group and LY294002 intervention group) were tested by Western blot.Except SB203580 intervention group and LY294002 intervention group,the fluorescence intensity in the cells of each group was analyzed with active oxygen detection kit and fluorescence microplate reader.Single factor analysis of variance and LSD test were performed for comparison between groups.Results Type Ⅰ collagen at the mRNA level of PDGF group (3.74±0.32) was higher than that of blank control group (1.00±0.00) ; Type Ⅰ collagen at the mRNA level of UA group (0.21 ±0.02) was lower than that of blank control group,UA intervention group (1.02 ± 0.12),DPI intervention group (1.09±0.21),SB203580 intervention group (1.18± 0.27),and LY294002 intervention group (1.15 ± 0.26) were all lower than PDGF group,and the differences were statistically significant (t =15.667,-4.501,-15.553,-15.154,-14.642 and -14.813,all P＜0.05).p47phox at the protein expression level of PDGF group (1.98±0.53) was higher than that of blank control group (1.00±0.00) ; that of UA group (0.48±0.10) was lower than blank control group; those of UA intervention group (0.95 ± 0.26),DPI intervention group (0.99 ± 0.28),SB203580 intervention group (0.93±0.31),and LY294002 intervention group (1.07±0.19) were all lower than PDGF group (t=4.209,-2.234,4.424,-4.252,-4.510 and-3.909,all P＜0.05).The protein expression level of PI3K of PDGF group (2.27±0.46) was higher than that of blank control group (1.00±0.00); that of UA intervention group (0.14 ± 0.07) was lower than PDGF group and blank control group; that of UA group (0.14±0.07) was lower than blank control group; those of DPI intervention group (0.53±0.25) and LY294002 intervention group (0.35±0.14) were all lower than PDGFgroup (t 6.205,8.208,-2.003,4.202,-8.502 and-9.831,all P＜0.05).The protein expression level of p-Akt of PDGF group (2.54±0.49) was higher than that of blank control group (1.00± 0.00); those of UA intervention group (0.74± 0.20),DPI intervention group (0.94 ± 0.37) and LY294002 intervention group (1.17±0.41) were all lower than PDGF group; that of UA group (0.59± 0.15) was lower than blank control group (t=5.927,-6.928,-6.158,-5.273 and-1.578,all P＜ 0.05).The protein expression level of p-p38MAPK of PDGF group (1.98±0.35) was higher than that of blank control group (1.00±0.00); those of UA intervention group (0.68±0.28),DPI intervention group (0.63±0.27) and SB203580 intervention group (0.67 ± 0.29) was all lower than PDGF group; that of UAgroup (0.28±0.13) was lower than blank control group (t=4.897,-6.479,-6.727,-6.529 and-3.561,all P＜0.05).The level of active oxygen of PDGF group (105.57±7.51) was higher than that of blank control group (69.60±8.63) ; those of UA intervention group (64.56±9.11),DPI intervention group (65.75 ± 6.62) was lower than PDGF group,UA group (29.84 ±3.19) was lower than blank control group (t=6.368,-7.288,-7.071 and-7.255,all P＜0.05).Conclusion UA could inhibit membrane displacement of NOX subunit p47phox and reduce active oxygen production in PDGF induced rat HSC-T6 cells,and then block phosphorylation of PI3K Akt,p 38MAPK signal pathways and inhibited the expression of type Ⅰ collagen at mRNA level.

Purpose To investigate the expression of EZH2 and p53 protein in breast cancer and to analyze their relationship with the clinical pathologic characteristics and prognosis. Methods The expression of EZH2 and p53 protein were detected by immunohisto-chemical method in 50 cases of breast adenosis tissues, 92 cases of breast invasive lobular carcinoma ( ILC) and 200 cases of breast in-vasive ductal carcinoma ( IDC) , and their correlation was also analyzed. Results There was no statistical significance of EZH2 be-tween ILC and IDC (P>0. 016 7), while its expression in breast adenosis tissues was lower than that in ILC and IDC (P<0. 016 7). In breast cancer the expression of EZH2 protein were not correlated with patient age, menopausal status, histological types, and pTNM stage. In contrast, its expression correlated with the tumor size, lymph node metastasis, molecular subtype, survival status and p53 (P<0. 05). There was no statistical significance of p53 between ILC and breast adenosis tissues (P>0. 016 7), while its expression in IDC was higher than that in ILC and breast adenosis tissues (P<0. 016 7). Its expression had no related to patient age, menopausal status, tumor size, lymph node metastasis in breast cancer, but related to histological types, pTNM stage, molecular subtype and sur-vival status (P<0. 05). The Kaplan-Meier survival analysis showed the expression of EZH2 and p53 had correlated with disease-free and overall survival rates of breast cancer (P<0. 05). Multivariate COX regression analysis showed that the expression of EZH2 and p53 were independent affecting factors to breast cancer patients. Conclusion The expression of EZH2 and p53 protein increase in the breast adenosis, ILC and IDC gradually, and they have positive correlation. The expression levels of EZH2 and p53 protein have im-portant value to evaluate the prognosis of breast cancer patients.

Purpose To investigate the clinicopathological characteristics and the survival outcomes of invasive lobular carcinoma. Methods A retrospective analysis of 98 patients with invasive lobular carcinoma and 530 invasive carcinoma of no special type was performed in order to observe the histological features and the clinical outcomes of invasive lobular carcinoma. Results Median follow-up was 68. 5 months for invasive lobular carcinoma and 67 months for invasive carcinoma of no special type. Invasive lobular carcinoma presented with a larger tumor size, more histopathological grade 2 tumors, increased rate of hormonal receptor positivity, human epider-mal growth factor 2 (HER-2) negativity, and had a lower proliferative index as compared to invasive carcinoma of no special type, more frequently presented with the luminal A subtype (P<0. 001). The classical invasive lobular carcinoma presented with a smaller tumor size, to have a lower histological grade and proliferative index compared to the non-classic type, and more frequently presented with the luminal A subtype, whereas the non-classic invasive lobular carcinoma patients more frequently presented with the luminal B, HER-2 overexpression, or triple negative subtype (P=0. 035). A statistically significant difference in the outcome was observed at un-ivariate analysis for patients with non-classic for disease-free survival (P=0. 043) and for overall survival (P=0. 048), as compared with patients with classical invasive lobular carcinoma. The disease-free survival difference between the invasive lobular carcinoma and the invasive carcinoma of no special type was not significant (P=0. 537), and the overall survival rates were not statistically different between the two groups (P=0. 397). A statistically significant difference of overall survival was observed at multivariate analysis for patients with HER-2 positive and triple negative subtypes versus patients with luminal A invasive lobular carcinoma (P=0. 015, P=0. 016) . Conclusions The outcome of invasive lobular carcinoma is significantly correlated with histological and immunohistochemi-cally defined molecular subtypes. New tailored strategies should be explored in these subgroups of patients with poor outcome.

Objective To explore the status of iodine nutrition in 8～10 years children after universal salt iodization(USI)in the iodine deficiency area.Methods Probability proportion to size method(PPS)or simple random sampling methods were used to sample children aged 8～10 years in iodine deficiency area in the vear 1995,1997,1999,2002 and 2005, respectively.Goiter were detected by palpation and B-ultrasound, iodine concentration in salt was detected by direct titration method and that in urine by the method of As3+-Ce4+catalytic spectrophotometry.Results After USI has been implemented,the median of salt and urinary iodine tended to mcreaseand the goiter rate tended to decrease year by year.In 2005,the coverage rate of iodinated salt was elevated to 97.2%,qualified iodize salt rate was 97.1%and edible qualified iodinated salt rate was 94.3%in the whole iodine deficiency areas.The median of urinary iodine Was 227.7 μg/L 89.7%(323/360)of the population had a level of the urinary iodine over 100μg/L Goiter rate of 8～10 years children Wag decreased from 22.3%(282/1267)to 4.4%(53/1200) from 1995 to 2005.Conclusion After 10-year USI,the status of iodine nutrition in ShaJldong Province has been promoted obviously and it is in a suitable iodine nutritional status.

Oroxylum indicum was a traditional Chinese medicine. In order to study the chemical constituents from the seed of O. indicum, the chemical constituents of 80% methanol extract of seeds of O. indicum were subjected to chromatography on silica gel, Sephadex LH-20, and preparative HPLC, leading to the isolation of eleven compounds. The structures were identified by various spectroscopic data including ESI-MS, 1H-NMR and 13C-NMR data as oroxin B (1), chrysin (2), baicalein (3), neglectein (4), quercetin-3-O-β-D-galactopy ranoside (5), quercetin-7-O-β-D-glucopyranoside (6), 2α,3β-dihydroxylluPeol (7), lupeol (8), rengyol (9), β-sitostero (10), and stigmasterol (11). Among them, compound 5 were firstly obtained from O. indicum.
Bignoniaceae ; chemistry ; Magnetic Resonance Spectroscopy ; Seeds ; chemistry

ObjectiveTo look into the distribution of “iodine suitable” region in iodine-deficient areas in Shandong province and to provide a scientific basis for guiding the redesignate of iodine-deficient areas and launch scientific supply of iodine.Methods One to 3 copies of water source samples were collected in 105 existing iodine-deficient counties by village.Water iodine content was detected by arsenic-cerium catalytic spectrophotometry.The areas with water iodine content below 10 μg/L was defined as iodine-deficient areas and among 10 - 150 μg/L were “iodine suitable areas” and greater than 150 μg/L were high iodine areas.Results The research was carried out in 14 cities,105 counties,and 1337 towns.We collected 65 716 water samples.Sample recovery efficiency reached 99.8％.The median of water iodine was 5.57 μg/L.In the 1337 towns surveyed,there were 82.05％(1097/1337) of the township with water iodine median ＜ 10 μg/L,17.43％(233/1337) between 10 - 150 μg/L,and 0.52％(7/1337) ＞ 150 μg/L.Conclusions In Shandong province,the water “iodine suitable” regions are distributed scattered with considerable proportion.In iodine-deficient areas,there are areas with high water iodine,and iodine-deficient regions should be redrawn.Emphasis should be put on iodine nutritional status of residents in “high iodine and iodine suitable” regions,and iodine supplementation should be carries out scientifically.

BACKGROUND/AIMS: To evaluate the enhancement patterns of liver metastases and their influencing factors using dynamic contrast-enhanced ultrasound (CEUS). METHODS: A total of 240 patients (139 male and 101 female; 58.5±11.2 years of age) diagnosed with liver metastases in our hospital were enrolled in this study to evaluate tumor characteristics using CEUS. A comparison of enhancement patterns with tumor size and primary tumor type was performed using the chi-square test. The differences between quantitative variables were evaluated with the independent-sample t-test and one-way analysis of variance. RESULTS: The enhancement patterns of liver metastases on CEUS were categorized as diffuse homogeneous hyperenhancement (133/240, 55.4%), rim-like hyperenhancement (80/240, 33.3%), heterogeneous hyperenhancement (10/240, 4.2%), and isoenhancement (17/240, 7.1%). There were significant differences in the enhancement patterns during the arterial phase based on the nodule size (p=0.001). A total of 231 of the nodules showed complete washout during the portal phase, and 237 nodules were hypoenhanced during the delayed phase. The washout time was correlated with tumor vascularity, with a longer washout time observed in hypervascular metastases compared to hypovascular metastases (p=0.033). CONCLUSIONS: Diffuse homogeneous hyperenhancement followed by rapid washout was the most common enhancement pattern of liver metastases on CEUS and was affected by the nodule size and tumor vascularity. Small metastases were prone to show diffuse homogeneous hyperenhancement. Hyper-vascular metastases showed a significantly longer washout time compared to hypovascular metastases.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Contrast Media/*therapeutic use ; Female ; Humans ; Liver/diagnostic imaging/pathology ; Liver Neoplasms/blood supply/*diagnostic imaging/secondary ; Male ; Middle Aged ; Neovascularization, Pathologic/diagnostic imaging ; Ultrasonography/*methods ; Young Adult

Objective:To observe the effect of electroacupuncture (EA) on the expressions of acetylcholine (ACh) and mucin 5AC (MUC5AC) in the lungs of rats with chronic obstructive pulmonary disease (COPD),and explore the mechanism of EA in treating COPD.Methods:Thirty Sprague-Dawley (SD) rats were randomly divided into a control group,a COPD group,and an EA group,with 10 rats in each group.The control group was a group of normal rats.The COPD rat model was induced by cigarette smoke combined with lipopolysaccharide (LPS).The COPD rats were treated with EA at bilateral Feishu (BL 13) and Zusanli (ST 36) in the EA group,30 min each time,once a day,successively for 14 d.The lung function was tested.The contents of ACh and MUC5AC in lungs and bronchoalveolar lavage fluid (BALF) were detected by enzyme-linked immunosorbent assay (ELISA).Pearson method was used to analyze the correlation between pulmonary function and the content of MUC5AC in lungs.The mRNA and protein expressions of MUC5AC in lung tissues were detected by real-time polymerase chain reaction (RT-PCR) and Western blot (WB),respectively.The immune response of MUC5AC was observed by immunohistochemistry.Results:Eight rats were left in each group,and the other two died.Compared with the control group,the total airway resistance (Raw) increased significantly and dynamic compliance (Cdyn) decreased significantly in the COPD group (P＜0.01);compared with the COPD group,the Raw level declined significantly and Cdyn increased significantly in the EA group (P＜0.01).The contents of ACh and MUC5AC in the lungs and BALF were remarkably higher in the COPD group compared with those in the control group (P＜0.01,P＜0.001);compared with the COPD group,the contents of ACh and MUC5AC were significantly lower in the EA group (P＜0.05,P＜0.001).There was a negative correlation between MUC5AC content and lung function (P＜0.001).The mRNA and protein expressions of MUC5AC in the lungs were significantly higher in the COPD group than in the control group (P＜0.001);compared with the COPD group,the expressions were significantly lower in the EA group (P＜0.01).Compared with the control group,the immune response of MUC5AC in the airway epithelium significantly increased in the COPD group (P＜0.001);the immune response of MUC5AC was significantly lower in the EA group compared with that in the COPD group (P＜0.001).Conclusion:EA treatment can improve the lung function of COPD rats,which may be related to its effect in the down-regulation of ACh and MUC5AC contents in the lungs as well as the inhibition of mucus hypersecretion.

OBJECTIVETo study the apoptosis inducing effects of bufalin on various human osteosarcoma cells and the concerning molecular mechanisms.

METHODMTT assay was used to detect the growth inhibition rates of osteosarcoma cells U-20S, U-20S/MTX300, SaOS-2, IOR/OS9 treated with bufalin in different concentrations and times. The apoptosis of cells was observed flow cytometry 48 h following bufalin treatment. The proteomic techniques were used to separate and compare the treated and control groups 48 h after bufalin-incubation. Then, the proteomic results were validated by western blot.

RESULTBufalin inhibited the growth of human osteosarcoma cells U20S, U20S/MTX300 (methotrexate resistant cells), SAOS2, IOR/OS9 in a dose- and time-dependent manner. The 72 h IC50 were (37.43 +/- 4.1), (32.24 +/- 5.3) nmol x L(-1) in U20S,U20S/MTX300 cells,respectivly. Flow cytometry showed that the apoptosis cells were increased following bufalin treatment. The protein expression profile showed 24 differentiated expression proteins. Among these proteins, the level of an anti-apoptotic protein, heat shock protein 27 (Hsp27) decreased significantly and the result was then validated by western blot. Ectopic expression of Hsp27 could reduce the bufalin-induced apoptosis remarkably in U20S and U20S/MTX300 cells.

CONCLUSIONBufalin could inhibit the cell growth and induce apoptosis on human osteosarcoma cells. The effect of bufalin may be related to the joint intervention with multiple protein targets. Among them, downregulation of Hsp27 plays a critical role in the bufalin-induced apoptosis in human osteosarcoma cells.

Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Bufanolides ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Drug Screening Assays, Antitumor ; Gene Expression Regulation, Neoplastic ; drug effects ; Humans ; Osteosarcoma ; pathology ; Proteomics

OBJECTIVETo observe effect and mechanism of n-Butanol lysate of alcohol extracts from Actinidia rufa root (monomer of R6,R8).

METHODTunel, Wright's stain with Giemsa's stain dyeing, and Hoechst 33258-PI double dyeing assay were used to detect the apoptosis of SGC7901 tumor cells treated with R6, R8. The SGC7901 tumor cells were randomly divided into control group and two treatment groups administered 0.05 g x L(-1) R6, R8, respectively, for 72 h). FCM assay was used to detect the apoptosis. Agarose electrophoresis assay was used to detect DNA strand break of tumor cells and reveal anti-tumor action mechanism.

RESULTThe apoptosis percentage of the tumor cell in 24 h, 48 h, 72 h was (17.08 +/- 2.78)% , (29.68 +/- 2.96)%, (52.46 +/- 3.81)%; (14.75 +/- 2.14)%, (27.35 +/- 3.79)%, (45.64 +/- 5.24)%, respectively, for the treatment group, significantly higher than that in the control group (1.94 +/- 1.55)%, (2.78 +/- 1.84)%, (11.8 +/- 2.79)% (P < 0.01) by tunnel assay. Wright's stain with Giemsa's stain dyeing assay, Hoechst 33258-PI and FCM double dyeing assay showed same action. R6 and R8 had the effect of inducing the DNA histogram of tumor cells (P < 0.01).

CONCLUSIONThe anti-tumor mechanisms may be associated with inducing the injury of DNA and stimulating apoptosis.

Actinidia ; chemistry ; Apoptosis ; drug effects ; Cell Line, Tumor ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Flow Cytometry ; Humans ; Immunohistochemistry ; Plant Roots ; chemistry