BackgroundProliferative vitreo-retinal disease (PVD)is one group of ocular complications marked by the enhanced proliferation of various cells included retinal pigment epithelial (RPE) cells.Insulin-like growth factor-1 (IGF-1)and vascular endothelial growth factor(VEGF) are implicated in the aberrant cell proliferation and pathological neovascularization that characterizes PVD,but the signaling mechanism is unclear now. Objective This study was to explore the effect of IGF-1 on VEGF in cultured human RPE cells under the small hairpin loop RNA (shRNA) keeping hypoxia-inducible factor-1 α ( HIF-1 α) silencing. Methods Human retinas were isolated from 4 healthy male donors,and the RPE cells were harvested and cultured.The ceils were identified using anti-human keratin antibody.The third to fifth generation of human RPE cells were used in the experiment.One target site of HIF-1α mRNA was chosen by certain design principle,and shRNA was designed and synthesized by the target site and transferred into the cells in vitro,and then the cells were cultivated with 50 μg/L IGF-1 for 24 hours.The mRNA and protein expressions of HIF-1α and VEGF were detected by RT-PCR and Western blot respectively. Results Cultured human RPE cells showed the flat irregularly multangular shape,and 97％cells appeared the positive response for keratin.HIF-1α mRNA expression in human RPE cells was significantly lower in 50 μg/L IGF-1 group than the 0 pg/L IGF-1 group ( 1.49±0.18 vs 1.46±0.17 ) ( t =0.335,P =0.743 ),however,the expressing levels of HIF-1α protein( 1049.86±172.54 vs 0.00±0.00) and VEGF mRNA(0.95±0.15 vs 0.35±0.07) and VEGF protein (391.98±56.77 vs 214.36±37.15)were raised in the 50 μg/L IGF-I group compared with 0 μg/L IGF-1 group (t=16.098,9.935,6.928,P＜0.05).After the HIF-1α-specific shRNA was transferred into cultured RPE cells,the expressions of both HIF-1α mRNA and its protein significantly decreased in RPE cells under 50 μg/L IGF-1 concentration condition( F=68.679,89.904,P=0.000),moreover,the expression of VEGF mRNA and its protein were significantly lowed(F=21.770,6.205,P＜0.05). ConclusionsIGF-1 promotes the accumulation of HIF-1α protein and induce the expression of VEGF in human RPE cells,which probably play a pivotal role in the development of PVD.

Objective To observe the protein and gene expression of immunoglobulin binding protein (BiP) in the femur of fluoride-treated rats, and preliminarily study the possible role of endoplasmic reticulum stress in the pathogenesis of skeletal fluorosis. Methods Sixty Wistar rats were divided into 4 groups according to body weight, n =15. The control and low-calcium groups were fed with normal diet(0.79％ calcium) and low-calcium diet(0.063％ calcium), respectively, and both drank tap water(fluoride concentrations ＜ 1 mg/L). High-fluoride and coexpesure to low-calcium groups were fed with conventional feed (0.79％ calcium) and low-calcium diet (0.063％ calcium), respectively, and both drank tap water containing sodium fluoride (sodium fluoride concentration of 221 mg/L). During experimental period, rats were measured body weight once a week with a stand diet and water available ad libitum. The experiment lasted for 12 weeks. The immunohistochemical and reverse transcription polymerase chain reaction(RT-PCR) techniques were used to detect the protein and gene expression of BiP in the femur of fluoride-treated rats and control subjects. Results The bone mineral contents of high fluoride, lowcalcium and coexposure groups[(0.131 ± 0019), (0.097 ± 0.011 ), (0.083 ± 0.007)g/cm] were lower than those of the control group[(0.159 ± 0.029)g/cm, all P ＜ 0.05]; the bone mineral density of low calcium and coexpesure to fluoride group[(0.243 ± 0.018), (0.223 ± 0.022)g/cm2] was lower than that of the control group[(0.296 ± 0.046)g/cm2, all P ＜ 0.05]. The immunohistochemical staining showed that the anti-BiP antibody positive osteoblasts were significantly increased in the low calcium diet and coexposure to fluoride groups than that in the control, and coexposure to fluoride elevated the positive cells than that in only low calcium diet group. The mRNA expression of osteopontin(OPN) and osteocalcin(OCN) in coexposure to fluoride with low-calcium group(1.36 ± 0.20, 1.31 ±0.11 ) was higher than that of the control groups (0.82 ± 0.16, 0.85 ± 0.15, all P ＜ 0.05) ; moreover, OPN expression significantly increased in this group than that of the only high fluoride group (0.97 ± 0.29, P ＜ 0.05). The mRNA expression of BiP in the low calcium and coposure to fluoride group (1.38 ± 0.24,1.35 ± 0.12) was significantly higher than that of the control group ( 1.14 ± 0.06, all P ＜ 0.05 ). Conclusions Higher fluoride or coexposure to low calcium diet stimulates the gene and protein expression in rat femur BiP, indicating that varying degrees of endoplasmic reticulum stress is likely involved in the pathogenesis of rat skeletal fluorosis.

Objective To evaluate whether Loewenstein occupational therapy cognitive assessment (LOTCA) can be used in the diagnosis of postoperative cognitive dysfunction (POCD) by comparing it with mini-mental state examination (MMSE).Methods Thirty patients performed cardiac valve replacement surgery in our hospital from October 2010 to March 2011 were chosen; the cognitive function of these patients were neuropsychologically evaluated by MMSE and LOTCA 1 d before and 7 d after the surgery; the differences of accurately diagnostic rate on POCD,the scores and the time-consuming when MMSE and LOTCA were chosen were compared.Results Thirteen patients (43.33％) were diagnosed as having POCD by MMSE and 17 patients (56.67％) by LOTCA; the difference of accurately diagnostic rate on POCD was not statistically significant (x2=1.067,P=0.302).MMSE scores,LOTCA scores and scores of 4 subtests (motor praxis,visuomotor organization,operation of thinking,and attention and concentration) 7 d after the surgery was significantly decreased as compared with those 1 d before the surgery (P＜0.05).MMSE scores were highly correlated with LOTCA scores (γ=0.711,P=0.005).Conclusion LOTCA can be used in the diagnosis and research of POCD.It can do a more comprehensive assessment than MMSE in cognitive function with longer times.

Objective To assess the relationship between the BMI and the brain DAT, and the influence of BMI on the brain SPECT imaging with 99Tcm-TRODAT-1. Methods MRI and 99Tcm-TRODAT-1SPECT imaging were performed in 31 healthy volunteers(16 males and 15 females), and then the three-dimensional reconstruction of SPECT images were completed. Based on the MRI images, right striatum (RST) and the left striatum (LST) were drawn as ROI on the 4 most clearly consecutive transverse slices.The cerebellum (CB) was taken as the background reference area and the corresponding uptake ratios of ST/CB, LST/CB and RST/CB were calculated. The Pearson correlation tests for radio-uptake ratios (ST/CB, LST/CB, RST/CB), BMI and age were performed, Then multiple linear regression analysis using ST/CB as dependent variable and BMI and age as independent variables was performed. SPSS 15.0 was used in data analysis. Results The ST imaging was symmetrical. The radioactivity was higher in the ST front area than that of the back area. The average uptake ratios of ST/CB, LST/CB, RST/CB were 1.71±0.16,1.70 ± 0. 16 and 1.72±0.17 respectively, in which the three ratios of the female were 1.74 ± 0. 18, 1.71±0. 19 and 1.76 ± 0. 19 respectively and those of the male were 1.68 t 0. 14, 1.68 ± 0. 13 and 1.69± 0.15respectively. ST/CB, LST/CB and RST/CB were negatively correlated with patients'BMI (r = -0. 53,-0.57,-0.47, all P＜0.05). The ST/CB was negatively correlated with patients' age(r=-0.39, P=0. 03). The multiple linear regression analysis showed that the BMI was significant independent variable (β=-0.53, t= -3.36, P=0. 002). Conclusions TheSTDAT,evel may decrease as patients' BMI and age increase. Females' DAT level is slightly higher than males'. For ST DAT imaging, age, gender and BMI should be all taken into consideration.

Objective To study the clinical and imaging features of patients with bone metastases from breast cancer and identify the factors related to the incidence of bone metastases. Methods Three hundred and thirty-four patients with breast cancer were recruited into this study. Whole-body 99Tcm-methylene disphosphonate (MDP) bone scan, clinical staging, pathological, immunohistochemical and serological test results were analyzed retrospectively. χ2 test was used for statistical analysis. Results The incidence rate of bone metastases for patients with and without lymph node metastases was 71% (152/214) and 22. 5% (27/120), respectively (χ2 =72.80, P =0.000). The incidence rate of bone metastases from infiltrated non-specified and specified breast cancer was 69% (203/294) and 41.7% (5/12), respectively (χ2 =3. 97, P=0.046). Alkaline phosphatase (ALP) was elevated in 28.5% (51/179) and 14.9%(11/74) of patients with and without bone metastases, respectively (χ2 = 5. 25, P = 0.022 ). Carcinoembryonic antigen (CEA), carbohydrate antigen (CA) 15-3, CA125, CA19-9 increased in 68.7% ( 123/179) and 27.0% (20/74) of patients with and without bone metastases, respectively (χ2 = 37. 03, P =0. 000). Conclusions The incidence of bone metastases from breast cancer is correlated to pathological types of primary tumor and lymph node metastases. Bone metastases occurs more frequently in patients with infiltrated, non-specified, primary cancer and with lymph node metastases. Serum ALP, CEA, CA15-3,CA125, CA19-9 might be the tumor makers for early diagnosis of bone metastases from breast cancer.

Objective To investigate the process of electrical activities when human brain deals with the facial sexual information. Methods Forty healthy college students in March 2005 were selected from Southern Medical University. All subjects participating in the experiment vohmtarily were right-handed with normal or corrected sight and never suffered from family history of mental disorder. 360 pictures of real human face (balf females and half males) strange to all participants, were used as the stimulus presented once one by one in randomized order on the screen with a stimulus duration of 800 ms and a stimulus onset asynchrony (SOA) of 1200 ms. Half subjects were asked to press the left button of a game-pad immediately after female face presentation and the right for male, the other half reversed. Event-related electroencephalogram (EEG) was recorded by 19 channels of international 10-20 system with linked earlobes as reference. EEG epochs from 100 ms before to 800 ms after stimulus were amplified by means of an ERPs system developed in our lab, digitized with sampling frequency of 1000 Hz, bandwidth of [0.1, 30] Hz and a notch of 50 Hz. Electrode impedance was less than 5 kΩ. Trial contaminated with ocular, muscular or any other type of artifacts were inspected visually and rejected. Each 60 stimuli of the same gender faces worked as an overlaying unit (the real overlay number was about 45-50 with bad epochs rejected). Three female and 3 male subjects were excluded owing to the bad ERP record quality and thus 34x6 epochs came from the rest ones. Results (1) The possible difference trend (but P>0.05) between male and female facial stimuli appeared at the frontal area in 40-100 ms after stimuli; (2) The significant difference between male and female appeared at the occipital in 140-160 ms after stimuli (P<0.05); (3) The significant difference between male and female appeared at the large frontal and occipital area in 200-260 ms after stimuli (P<0.05); (4) The significant difference between male and female appeared at the large central parietal in 280-300 ms after stimuli (P<0.05); (5) The significant difference or trend between male and female appeared at several areas in different time after 360 ms. Conclusion In different stage, different brain areas are activated for the facial sexual feature processing. Thus, our brain works as a network.

Objective To investigate the relationship between the patient-based questionnaires and the computed tomography (CT) staging in patients with chronic rhinosinusitis (CRS). Methods Quantitative data of 121 preoperative recruits with CRS were collected by using the Lund-Mackay CT staging system, a visual analogue scale ( VAS), sino-nasal outcome test-20 ( SNOT-20), and the medical outcome study short-form 36 items (SF-36). The patients were classified into several subgroups according to whether CRS was associated with nasal polyps (NP) or not, sex, duration of disease, and educational background.Correlation between the patient-based questionnaires and the CT staging were analyzed in the total cohort patients and subgroups. Results In the total cohort patients, there were significant correlations between SNOT-20 and SF-36 ( r = -0. 561, P ＜ 0. 01 ), SNOT-20 and VAS ( r = 0. 743, P ＜ 0. 0l ), and SF-36 and VAS ( r = - 0. 504, P ＜ 0. 01 ), however, the CT staging did not correlate with the patient-based questionnaires (P ＞ 0. 05 ). Significant but weak correlations were found between the CT staging and the patient-based questionnaires in the C RS with NP subgroup (CT vs SNOT-20, r = 0. 318, P = 0. 005; CT vs SF-36, r = - 0. 358, P = 0. 002; CT vs VAS, r = 0. 358, P = 0. 002). Compared between CRS with NP and without NP subgroup, there were statistic differences on the Lund-Mackay CT stage and the SNOT-20 and VAS scores (t value was 3.249, -2.409, -2.957, respectively, all P＜0.05). Conclusions The patient-based questionnaires correlate well with each other, but CT staging correlated significantly but weakly with the patient-based questionnaires only in the CRS with NP subgroup. Nasal polyps do not appear to be responsible for the adverse effects of CRS on quality of life.

Objective To explore the mechanism of fine particulate matter (PM2.5) induced endothelial injury and the efficacy and mechanism of ginsenoside Rg1 on the inhibition of endothelium injuries in human endothelial cells exposured to PM2.5.Methods Human umbilical vein endothelial cells (HUVECs) were stimulated with various concentrations PM2.5 (0.1,0.2,0.4,0.8 mg/ml) and PM2.5 at concentration 0.8 mg/ml induced significant endothelial injury and was chosen for the main study in the presence or absence of Rg1 (0.04 mg/ml).After 24 h treatment,cell growth A value was detected through MTT,intracellular reactive oxygen species (ROS) level through fluorescence labeling probe method and HO-1,Nrf2 mRNA expression was detected by RT-PCR.Results The cell A value was significantly lower while the ROS fluorescence gray value and the average optical density ratio of HO-1 were significantly higher in PM2.5 group than in the control group (all P ＜ 0.05).The average optical density ratio of Nrf2 was similar between PM2.5 group and control group (P ＞ 0.05).The A value and the average optical density ratio of HO-1 were significantly higher while the ROS fluorescence gray value was significantly lower in co-treated PM2.5 (0.8mg/ml) + Rgl (0.04 mg/ml) group than in the PM2.5 (0.8 mg/ml) group (all P ＜ 0.05).Conclusion PM2.5 could induce human endothelial cells injury by increasing oxidative stress which could be attenuated by ginsenoside Rg1.

Background Bone morphogenetic protein-6 (BMP-6) is closely correlated with tumor differentiation and skeletal metastasis. Estrogen is considered as a stimulant for the initiation and promotion of breast cancer. Previous studies demonstrated that 17β-estadiol (E2) can selectively increase the expression of BMP-6. This experiment is designed to detect the molecular mechanism of estrogen activating BMP-6 gene transcription in human estrogen receptor positive (ER+) breast cancer cell line MCF-7. Methods After the treatment of MCF-7 cells with E2 at different concentrations (10-11 mol/L, 10-9 mol/L, 10-7 mol/L), the BMP-6 expression level was examined through real-time polymerase chain reaction. Through restriction enzyme digestion, human BMP-6 1.2 kb long promoter, BMP-6 0.7 kb long promoter was cloned into pGL-3 basic vector; after the treatment with 10-7 mol/L E2, luciferase activities of the two promoters were detected. Site-directed mutagenesis was performed to obtain the mutant forms of estrogen response element half-site (1/2 ERE) element and Sp1 sites in the BMP-6 promoter, the activities of these mutant form promoters were detected following the methods mentioned above. Chromatin immunoprecipitation (ChIP) assay was also used to confirm the binding of estrogen receptor α (Erα) on BMP-6 promoter in the presence of E2. Results E2 dose dependently increased BMP-6 mRNA expression in human ER+ breast cancer cell line MCF-7. At a dose of 10-7 mol/L E2, human BMP-6 1.2 kb promoter activity was increased by 90% compared with the control group treated with ethanol (P<0.05). Both the 1/2 ERE response element mutant form and the Sp1 site mutant form of the BMP-6 promoter abolished the activation of the BMP-6 promoter's response to E2. Through ChIP assay, the binding of Erα on 1/2 ERE response element in BMP-6 promoter was further validated. Conclusion Estrogen induces BMP-6 expression in human ER+ breast cancer cell line MCF-7 through its receptor Erα binding on 1/2 ERE element in the BMP-6 promoter.