BACKGROUND: Puerarin possesses various biological efficacies, such as the protective efficacy on hypertension, cardiac disease, diabetes mellitus and blood disease, and the extracts of puerarin can inhibit the proliferation of S180 sarcoma and Lewis lung cancer to some extent.OBJECTIVE: To investigate the modulation effects of total flavone of puerarin (TFP) on the growth of pheochromocytoma PC12 cells and the protective efficacy on the H2O2-induced cellular oxidative damage.DESIGN: Complete randomization design and control experiment.SETTING: Department of Biochemistry and Institute of Geriatrics, Chinese PLA General Hospital.MATERIALS: Puerarin was bought from Tongrentang drug store, and TFP was extracted and purified routinely by ethanol and ether acetate, then was evaluated with thin-layer chromatography. The content of puerarin in extracts was 31.79% in quantitative assay. Methyl thiazolyl tetrazolium (MTT), luminal and anti-oxidative activity reagent xanthine oxidase were all from Sigma Company. PC12 cells were given by Institute of Geriatrics,Chinese PLA General Hospital as a present.METHODS: The experiment was conducted at the Institute of Geriatrics, Chinese PLA General Hospital from January to July in·2001.① The cells were cultured in 20 g/L DMEM (pH 7.1-7.2), and randomized into two groups: TFP group and H2O2-injury+TFP group, and each group was divided into 5 mass concentrations (0, 1.0, 10, 100 mg/L and 1.0 g/L). There were 8 holes for parallel culture with 100 mL culture medium in each hole (containing 1 ×l09 L-1 cells). TFP group:TFP was added for 72-hour culture at 37 ℃; H2O2 injury+TFP group:TFP was firstly cultured for 48 hours at 37 ℃, then 500 mmol/L H2O2 was added and co-cultured for other 24 hours.②The activity of cultured PC12 cells was monitored by MTF assays, the content of nitrite was measured by Griess reagents, and the antioxidant activity of superoxidedismutase (SOD) was monitored by hypoxanthine/xanthine oxidase.H2O2-initiated PC12 cellular oxidative damage had been used as experimental model to study the protective efficacy of TFP, and expressed as inhibition ratio [(blank A value-detection A value)/blank A value × 100%]. The higher inhibition ratio indicated the strong ability of clearing O2-. ③ One-factor analysis of variance was used to compare the difference between data. MAIN OUTCOME MEASURES: The influence of TFP on the nitrite content, SOD activity and cell activity in PC12 cells.RESULTS: ①Effect of TFP on PC12 cell activity: 1-10 mg/L TFP hadno obvious effects on the growth of PC12 cells, and 100 mg/L TFP in creased the cell growth (P ＜ 0.05), whereas the TFP concentration was increased to 1.0 g/L, the activity of PC12 cells was inhibited obviously (P ＜ 0.01). TFP of 1-100 mg/L could protect the cultured cells from the oxidative damages of H2O2 concentration dependently (P ＜ 0.05).② Effect of TFP on clearing O2-: The ability of clearing O2 increased withthe mass concentrations of TFP in both groups with obvious dose-effect relation, except when 1 mg/L TFP was added in the H2O2 injury+TFP group. The SOD activity in PC12 cell culture liquid was obviously en hanced after adding 100 mg/L and 1 g/L TFP, compared with that without TFP addition (P ＜ 0.05-0.01). ③Modulation of TFP on nitrite: TFP of low concentration (1-100 mg/L) reduced the production of cellnitrite, whereas increased the nitrite production at the concentration of1.0 mg/mL.CONCLUSION: ①TFP can regulate the growth of PC12 cells, which canbe enhanced by low-concentration (1-100 mg/L) TFP whereas inhibited byhigh-concentration (1 g/L) TFP. However, the anti-oxidation of TFP is themost powerful. ②TFP can protect the PC12 cells obviously from the oxidative damages induced by H2O2 at low concentration.

Objective To evaluate efficacy in detecting lung nodules at low-dose CT(LDCT) by nodule enhanced viewing(NEV).Methods One hundred and twenty seven patients who were referred to undergo low-dose CT (LDCT) for the evaluation of pulmonary metastasis or screening lung cancer were selected randomly.Two radiologists with at least 10 years experience read the images with normal clinical reading speed to find actionable nodules ≤ 2.0 cm in maximum diameter,and their consensus result was referred as Standard.NEV was adopted to detect the pulmonary nodules.Two residents with experience of less than three years read first detected suspicious nodules and recorded reading time,first consensus and mean time were recorded.Then,they made second decisions on the images with the help of NEV and the results and the reading time were recorded and analyzed by using wilcoxon test.The sensitivity and accuracy of NEV,residents and residents with NEV were analyzed.Results Standard,resident,NEV and resident with NEV detected 570,404,768 and 593 lung nodules ≤2.0 cm in maximum diameter,respectively.More than 60％ nodules were less than 0.5 cm in maximum diameter.The performance of NEV in detecting nodules ≤2.0 cm as well as nodules ＜ 0.5 cm in maximum diameter was significantly higher than that of the resident(Z =-6.887,P ＜0.01 and Z =-7.235,P ＜0.01),and the performance of resident with NEV indetecting nodules ≤2.0 cm as well as nodules ＜ 0.5 cm in maximum diameter was significantly higher than that of resident without NEV (Z =-6.606,P ＜ 0.01 and Z =-6.657,P ＜ 0.01).The resident,NEV and the resident with NEV detected nodules ＜ 20 mm in maximum diameter with sensitivities of 61.4％,86.3％ and 95.3％,and with accuracy of 56.1％,58.1％ and 87.6％,respectively.The resident achieved sensitivities of 51.4％,88.1％ and 94.8％,and accuracy of 47.0％,56.9％ and 87.5％ for nodules ＜5 mm in maximum diameter,respectively.The resident,NEV and resident with NEV spent 120-444 s,85-262 s and 131-1512 s per case to read the CT scans,respectively.The reading time of resident with NEV in was significantly higher than that of resident without NEV(Z =-9.781,P ＜ 0.01).The resident spent 23 s per NEV mark.Conclusion NEV considerable improves the resident's performance in lung nodule detection,especially in maximum diameter ＜ 0.5 cm nodule detection.

Objective The purpose of this study was to examine the inhibitory effect of ONO-AE3-208, an EP4 antagonist, on prostate cancer with bone metastasis in an animal model . Methods A PC3/LUC cell line was constructed by stably transfecting luciferin to prostate cancer PC 3 cells and inoculated into the left ventricle of nude mice to establish an animal model of prostate cancer with bone metastasis .After modeling , the animals in the experimental group and control groups were intraperitoneally given ONO -AE3-208 and double-distilled water, respectively, followed by examination of the metastasis loci and tumor burden by bioluminescence ima -ging and statistical analysis with survival curves . Results At 60 days after modeling , the animals in the control group exhibited sig-nificantly increased metastases and fluorescence burdens as compared with the experimental group (P<0.01), and the increase was in a time-dependent manner (P<0.01).At 60 days, the controls began to die while the experimental animals remained well alive , and at 180 days, the mice of the control group all died .The survival rate of the animals was significantly higher in the experimental group than in the control ( 13.3% vs 0%, P <0.01 ) and the median survival time remarkably longer in the former than in the latter group (162 d vs 116 d, P <0.01). Conclusion The EP4 antagonist ONO-AE3-208 inhibited the bone metastasis of prostate cancer and prolonged the survival time in the model mice .

Objective To determine the effect of chronic restraint stress during the late pregnancy on the behavior parameters of learning and memory of the rat offspring. Methods Stress was inflicted each day of pregnancy from day 13 to day 20. Pregnant females were individually restrained for 45 min three times a day. After delivery, one-month-old and three-month-old female and male rats were examined in Morris water maze to analyze the effect of exposure to prenatal restraint stress on the spatial learning and memory. Results In comparison to the control group, the latency to find the platform in the prenatally stressed group was significantly prolonged in both one-month-old and three-month-old offsprings (P

Objective To study the effect of chronic restraint stress on the expressions of Bax, Bcl-2, Fas and FasL in hippocampus of the offsprings during the pregnancy. Methods Restraint stress was performed three times each day of pregnant women from day 13 to day 19 (each time for 45min). After delivery, the expression of Bax, Bcl-2, Fas and FasL in hippocampus of one-month-old female and male rats were respectively examined by immunohistochemical method. Results Compared with the control group, the expression level of Bax in stressed group is higher (P 0.05). Conclusion Restraint stress during the pregnancy can upgrade the expression of Bax and degrade the expression of Bcl-2 of the offsprings; no effect was found on the expressions of Fas and FasL of the offsprings.

Objective Present study aimed to research the mechanism of the use of 0.2% brimonidine tartrate eye drops in the treatment of optic nerve crush injury of rat.MethodsAnimal models of optic nerve crush injury were created in 60 SD female rats by clipping the exposed optic nerve at 2 mm in retrobulbar for 6 seconds with 78 grams of reverse forceps.The successful model was identified as Marcus-gun pupil without bleeding of fundus after operation.The animals were randomly assigned to model group and brimonidine treating group,and another 30 normal SD rats were used as the normal control group.The 0.2% brimonidine tartrate eye drops was topically administered at 2 hours before operation and after operation twice per day in the brimonidine treating group.The retinas from 18 rats were isolated after 3,7,21 days for RGCs counting by H&E staining,and the retinal ultrastructure was examined under the transmission electron microscope.The retinas from the other 72 SD rats (including normal,model and brimonidine groups) were prepared for the detection of bcl-2 and bax using immunohistochemistry l,3,5,7,14,21 days after the operation.ResultsNormal and almost normal retina structure was exhibited in rats of the normal group and brimonidine treating group,but disorder of cellular arragement and decrease of retinal thickness were found in the model rats under the optical microscope.The RGCs counting was significantly different among the three groups from 3 days through 21 days after operation with the considerably declination in the model group and brimonidine treating group compared with the normal control group (P＜0.05-0.01).However,that of the brimonidine treating group was obviously increased in comparison with the model group (P＜0.01).The expression of bax in rat retina was obviously reduced (P＜0.01),but the expression of bcl-2 was increased in brimonidine group compared with the model group from 5 through 7 days after operation (P＜0.01).ConclusionThe 0.2% brimonidine tartrate eye drops has a preventive effect on optic nerve crush injury of rat,and its inhibition on apoptosis is one of the mechanisms.

Objective Wnt signaling plays an important role in the development and progression of renal cell carcinoma (RCC).This study aimed to evaluate the effects of the Wnt signaling inhibitor 15-oxospiramilactone on the proliferation , migration, cell apoptosis, and cycles of the human RCC cell line 786-0, and to investigate the possible mechanisms of this small molecule acting on RCC in ivtro. Methods We treated 786-0 cells with DMSO ( blank control group ) and 15-oxospiramilactone at the concentrations of1.25μmol/L (low 15 -OSL), 2.5μmol/L (medium 15-OSL), and 5μmol/L (high 15-OSL), respectively, for 72 hours.Then we observed the changes in the proliferation and migration of the 786-0 cells by MTT and scratch-wound assay and determined their apopto-sis and cycles by Annexin V-FITC/PI assay and flow cytometry . Results 15-oxospiramilactone significantly inhibited the growth of the 7860-cells, with the IC 50of 1.088 μmol/L at 72 hours, and decreased their migration distance (P<0.05).After 36 hours of treatment, the apoptosis rates of the 786-0 cells in the low, medium, and high 15-OSL groups were (12.17 ±0.56), (18.54 ± 1.07), and (50.74 ±1.28) %, respectively, significantly increased as compared with (7.85 ±0.42) %in the blank control group (P<0.05), and in an obviously concentration-dependent manner.15-oxospiramilactone remarkably reduced the number of cells in the G0/G1 phase and increased that in the G 2/M phase (P<0.05). Conclusion 15-oxospiramilactone can significantly inhibit the pro -liferation and migration and induce the apoptosis of 786-0cells in vitro.It may be a potential anti-RCC agent.

Objective To investigate the correlation between the paired inspiratory and expiratory quantitative CT and pulmonary function tests (PFTs) in patients with chronic obstructive pulmonary disease (COPD).Methods A total of 84 patients with COPD were enrolled.For each patient,CT scan was performed in deep inspiration and expiration.Using automatic post-processing software,a three-dimension quantitative measurement was employed to assess the CT parameters of emphysema and air trapping.The correlation between CT and PFT was evaluated by the Spearman rank correlation test and multivariate linear regression analysis.Results The percent 1 [Perc1,(-984.28 ± 17.93) HU] and percent 15 [Perc15,(-948.35 ± 22.26) HU] from the CT parameters of emphysema were positively correlated with the forced expiratory volume in 1 second predicted (FEV1％,48.69 ± 23,47),the ratio of forced expiratory volume in 1 second over forced vital capacity [FEV1/FVC,(45.89 ± 15.36)％,r =0.454.-0.67,P ＜0.01],was negatively correlated with the ratio of residual volume to total]ung capacity [RV/TLC,(61.32 ±14.48)％].The other CT parameters of emphysema index (EI) and the parameters of air trapping,the change in relative lung volume with attenuation values from-860 to-950 HU [RVC-860--950,(17.66 ±22.36)％],the expiration to inspiration ratio of mean lung density (MLDex/in,0.93 ± 0.06),the expiration to inspiration ratio of lung volume(LVex/in,0.71 ± 0.14) had negative correlations with logFEV1 ％,FEV1/FVC (r=-0.48--0.69,P＜0.01) and positive correlations with RV/TLC (r =0.41-0.66,P ＜0.01).The further univariate linear regressions showed that EI,Perc1,Pere15,RVC-860--950,MLDex/in,LVex/in were correlated with the parameters of PFTs (R square values of the regression equation,ranged from 0.27 to 0.66,P ＜ 0.01).After the pairwise combinations of the parameters of emphysema and air trapping,multivariate stepwise regressions showed better R square values from 0.66 to 0.85.Conclusions CT parameters of emphysema and air trapping can reflect pulmonary function in COPD patients.The combinations of the two types of CT parameters are more correlated with the PFTs parameters.

Objective To evaluate the relationship between the lung volume (LV) in inspiration and expiration,pulmonary function tests(PFT),and other CT measurements of emphysema index (EI) and mean lung density (MLD) in patients with chronic obstructive pulmonary disease (COPD).Methods Seventy-six patients with COPD were included.Three-dimension analysis was performed to obtain the following CT parameters on the inspiratory and expiratory phases:EI,MLD,LV.The ratios and differencesof MLD and LV between the two phases were calculated(ΔMLD,ΔLV,MLDex/in,LVexin).Not only the linear correlations between the lung volume parameters and PFT but also the correlations of lung volume parameters with the other CT parameters were tested by the Spearman rank correlation test and multivariant step wise regression.Results LVex/in had negative correlations with forced expiratory volume in 1 second ％ predicted [FEV1 ％,(54.32 ± 7.11)] and the ratio of forced expiratory volume in 1 second to forced vital capacity [FEV1/FVC,(49.12 ±8.01)％] (r =-0.69,-0.56,P ＜0.01),but it had the positive correlation with the ratio of residual volume to total lung capacity [RV/TLC,(58.03 ± 8.55) ％,Spearman coefficients 0.66,P ＜ 0.01].LV/exin (0.67 ± 0.12) was positively correlated with MLDex/in (0.89 ± 0.04,r =0.88,P ＜0.01).The further multivariate step wise regression denonstrated that EI,LV and MLD could introduce a regression equation with R2 =0.77 and 0.73,respectively (P ＜ 0.01).Conclusions There is an association between LVex/in and the parameters of routine PFT,which can reflect the collapsibility of lung.Moreover,LVex/in can be considered to be equivalent to MLDex/in.Taking into account the impact of scanning parameters on MLDex/in,LVex/in may play a complementary role in the assessment of pulmonary function.

Objective Based on the diagnostic technique of electric meridian detection, to observe the clinical efficacy of acupuncture in treating angioneurotic headache.Method Totally 106 patients with angioneurotic headache were randomized into a treatment group and a control group, 53 cases in each group. The treatment group received acupuncture based on the electric meridian detection, while the control group received medication treatment. The hemorheology indexes [plasma viscosity (PV), erythrocyte sedimentation rate (ESR)] and McGill Pain Questionnaire (MPQ) [Pain Rating Index (PRI), Visual Analogue Scale (VAS), and Present Pain Intensity (PPI)] were observed before and after intervention, and the clinical efficacies were compared. Result The total effective rate was 88.7% in the treatment group versus 71.7% in the control group, and the between-group difference was statistically significant (P<0.05). After intervention, the hemorheology indexes and MPQ scores were significantly changed in both groups (P<0.01,P<0.05).Conclusion Acupuncture based on electric meridian detection is an effective method in treating angioneurotic headache.