The purpose of this paper is to investigate the reversal effect of small interfering RNA (siRNA) targeting MDR1 and MDR3 genes on the resistance of MCF-7/ADR cells to adriamycin. siRNA plasmid vector targeting MDR1 and MDR3 genes was transfected into MCF-7/ADR cells, and then was stained with Annexin-V FITC (fluorescein isothiocyanate conjugated) to detect the early stage cell apoptosis by flow cytometry (FCM). 50% inhibition concentration (IC50) of adriamycin for MCF-7/ADR cells was determined by MTT method. MDR1 and MDR3 mRNA was assessed by RT-PCR. Treatment of MCF-7/ADR cells with the two kinds of siRNAs resulted in a reversal of adriamycin resistance of MDR to different extents. 1) The apoptosis efficiency of MDR1 and MDR3 siRNA vector after transfection was (18.21+/-1.65) % and (9.07+/-2.16) % respectively (P<0.05), and there was significant differences in the apoptosis efficiency between pSuppressor Neo vector and the MDR1siRNA or MDR3 siRNA vector (P<0.01); 2) The reversal effect of MDR1 siRNA is higher than that of MDR3 siRNA (P<0.05); 3) The expression of MDRI and MDR3 mRNA can be restrained by pSuppressor Neo MDR1 and MDR3 siRNA respectively, and the reduction in the mRNA level was in a time-dependent manner (P<0.01). MDR1 and MDR3 gene silencing can enhance intracellular adriamycin accumulation in MCF-7/ADR cells, improve sensitivity of MCF-7/ADR cells to adriamycin, and induce cell apoptosis. The reversal effect of adriamycin resistance by siRNA of MDR1 was more effective than that of MDR3.

To explore the role and possible mechanism of apoptosis and caspase-3 activity in the development of multicellular drug resistance of ovary cancer. Ovarian cancer cell A2780 multicellular spheroids (MCS) were obtained from three-dimensional culture. Drug sensitivity of monolayer cells (MC) and MCS were respectively tested by MTT staining and cytometry. The apoptosis of MC and MCS were determined by the flow cytometry (FCM). The expression of bcl-2 and caspase-3 in A2780/MC and A2780/MCS were detected by using Western blot and caspase-3 assay kit. A2780/MC was compacted into mass after 2 days in three-dimensional cell culture model, and MCS had more than two layers of cells growing within 5 days. Compared with A2780/MC, A2780/MCS were more resistant to the anticancer drug, and the apoptosis rate was significantly lower than those of A2780/MC. The activity of caspase-3 in A2780/MCS was significantly lower than the A2780/MC. But the expression of bcl-2 in A2780/MCS was significantly higher than that in A2780/MC. It was suggested that the drug resistance of MCS might be associated with the overexpression of anti-apoptosis protein bcl-2 and the down-regulation of caspase-3 activity.

To investigate the relationship between the expression of early growth response gene 1 (EGR-1) and p38MAPK pathway in the paclitaxel resistance of ovarian carcinoma cells, the effect of p38MAPK inhibitor SB203580 on cell apoptosis was examined by using Hoechst 33258 staining. The intracellular Rh123 (Rhodamine 123) accumulation was detected by the flow cytometry (FCM). The 50% inhibition concentration (IC50) of paclitaxel for A2780/Taxol cells was determined by MTT method. Electrophoretic motility shift assay (EMSA) was employed to examine the EGR-1DNA binding activity. MDR1 and EGR-1 mRNA were assessed by RT-PCR. The expressed of p-gp, phosphorylated p53 and p38 were detected by Western blotting. SB203580 could remarkably promote the apoptosis of A2780/Taxol cells, and the cell apoptosis was in a time-dependent manner. Cellular Rh123 accumulation was increased, and the IC50 of paclitaxel for A2780/Taxol cells was decreased significantly. A2780/Taxol cell line after SB203580 treatment was shown to have a significantly higher level of EGR-1 DNA binding activity. SB203580 down-regulated the activity of p38MAPK pathway, but up-regulated EGR-1 expression. SB203580 significantly increased the level of cellular phosphorylated p53 protein, but decreased the p-gp protein level and MDR1 mRNA level in A2780/Taxol cells. There existed a close relationship between p38MAPK pathway and the paclitaxel resistance of ovarian carcinoma cells. The expression of EGR-1 mediated by p38MAPK pathway plays a critical role in paclitaxel resistance of ovarian carcinoma cells.

BackgroundHigh myopia is one of leading causes of blindness,so far the pathogenesis remains unclear.Two single-nucleotide polymorphisms (SNPs) of rs6885224 and rs12716080 in CTNND2 gene were recently found to be associated with high myopia in Singaporean Chinese.But whether these SNPs are related with the pathogenesis of high myopia in Han Chinese is worth studying,Objective This study was to investigate the relationship between the genetic variations of the CTNND2 gene and high myopia in Han Chinese.MethodsA case-controlled association study was designed.Nine hundred and thirty-three individuals with high myopia and 1227age- and gender-matched normal subjects were included in this study.The 5 ml of periphery blood was obtained from all subjects for the extraction of genomic DNA.The target DNA was amplified using PCR and purified by the SNaPshot method.Four SNPs rs12716080,rs917012,rs6885224 and rs16901340 in the CTNND2 gene were genotyped.This study was approved by the Ethic Committee of Sichuan Provincial People Hospital.Written informed consent was obtained from each subject before his/her enrollment.Results The frequencies of the genotypes rs6885224,rs12716080,rs917012,rs16901340 SNPs were in Hardy-Weinberg equilibrium (HWE) ( P=0.181,0.085,0.732,0.313,0.264,0.663,0.084,0.196).There were no significant differences in genotypes frequency distribution ( in turn P =0.654,0.406,0.828,0.403 ) and allele frequency distribution of the CTNND2 gene ( in turn P =0.377,0.209,0.743,0.198) between the high myopia group and normal control group.The haplotypes (TA and GA)frequencies of rs12716080 and rs917012 in the high myopia group were significantly different from those of the normal control group(TA:0.784 vs.0.719;GA:0.087 vs.0.136) (x2 =6.115,P=0.013 ;x2 =6.634,P=0.010),but those of GG were similar between the high myopia group and normal control group ( 0.123 vs.0.143,x2 =0.889,P =0.346). ConclusionsThe SNPs rs12716080,rs917012,rs6885224 and rs16901340 in CTNND2 gene were not responsible for high myopia,however,the haplotypes of rs12716080 and rs917012 are susceptible for high myopia in Han Chinese.

The changes of some parameters of Gray type Ⅰ synaptic interface in the brain of mice treated with desglycinamide-arginine-8-vasopressin (DGAVP) have been analysed quantitatively two hours after DGAVP injection. The animals were killed and the hippocampus and sensori-motor area of cerebral cortex were prepared for electron microscopy. The electron microphotographs were analysed by IBM-PC computer image processing system. The main results of the experiment are as follows:1. The thickness of postsynaptic density apparently increased in both the sensori-motor area of cerebral cortex and CA_3 area of hippocampus after injection of DGAVP (P

The biotechnology of glycolipid fermented b y a bacillus coagulans was studied and the fermentation pro cess in 10L bioreactor was conducted.Suitable medium contained 6% bean oil as ca rbon source,3 5g/L NaNO 3 as nitrogen source,0 75g/L yeast extract and some i no rganic salts.The fermentation temperature of 30℃,initial pH of 8 5,strring rev olution of 150～240r/min and fermentation period of 96h proved to be optimal.The yield of glycolipid in 10L bioreactor was 7 073g/L.

Objective To comment the severity of severe hand,foot and mouth disease(HFMD)by pediatric risk of mortality score(PRISM),and assess the performance of PRISM in predicting mortality or complication probability in HFMD.Methods Four hundred and twenty-four severe HFMD pediatric patients were recruited in the study from 1th Jan 2010 to 31th June 2013.Information on the outcome and the varia-bles required to calculate PRISM score were collected.The logistic regression model developed in the learning sample was evaluated in the test sample by calculating the area under the receiver operating characteristic (ROC)curve to assess discrimination pneumorrhagia and death.Calibration across deciles of risk was evalua-ted using the Hosmer-Lemeshow goodness-of-fit χ2 test.Results The area under the ROC curve were 0.87 (95%CI 0.80~0.94 )for PRISM in predicting pneumorrhagia probability.The area under the ROC curve were 0.87(95%CI 0.80~0.95)for PRISM in predicting mortality probability.The PRISM in observed and expected pneumorrhagia did not demonstrate good calibration at ten mortality risk intervals (χ2 =36.66, P<0.001 ).The PRISM in observed and expected mortality did not demonstrate good calibration at ten mortali-ty risk intervals(χ2 =41.11,P<0.001).Conclusion The PRISM score is demonstrated good discrimination of pneumorrhagia and death in HFMD pediatric patients,but the performance of calibration is not good.

Background and purpose: Astrocytoma is the most common neuroepithelial neoplasm, and its grading has profound effect on its treatment and prognosis. To investigate the application of stepwise discriminant analysis in grading astrocytomas, this study developed two models of stepwise discriminant analysis according to relevant factors of astrocytoma. Methods: From January 2008 to April 2009, 111 primary astrocytoma patients were enrolled. Each patient was scored based on location, signal intensity on T1WI, signal intensity on T2WI, enhancement, edema, border, cyst or solidness, and mass effect of their magnetic resonance images. With their age score of grading, Fisher stepwise discriminant analysis and the Logistic discdminant were used. The results from the two models were then evaluated and compared. Results: According to Fisher stepwise diseriminant analysis, the predictive accuracy was 87.7% with 80.0% sensitivity, 91.5% specificity and 0.942 area of ROC curve. However, the predictive accuracy of Logistic discriminant analysis was 84.9% with 80.0% sensitivity, 86.8% specificity and 0.940 area of ROC curve. There were no statistically significant differences in terms of accuracy (P=0.250) and areas under ROC curve (Z=0.433, P=0.665) between the two models. Conclusion: Two stepwise discriminant analysis models are meaningful to predict the grading of astrocytoms, and the application of Fisher stepwise discriminant analysis is simpler than the Logistic discriminant analysis.

OBJECTIVETo investigate and study the endangered Tibetan medicinal plant species, their moisture content, biomass and resources reserves in Lhasa region.

METHODThe rare and endangered Tibetan medicinal plant resources were investigated by plot-quadrat method, walking and inquiry ways, sampling and drying method.

RESULTThere were 37 species of rare and endangered plants, belonging to 22 families and 34 genera in Lhasa region. The moisture content of aerial part was higher than that of underground part in many plants. The moisture content of Przewalskia tangutica was the highest (91.97%), and the lowest one was Fritillaria delavayi (only 25.99%). The mean biomass of Rubus biflorus was the highest (1 830.480 g), that of Cordyceps sinensis was the lowest (0.291 g). The root-shoot ratio of Asparagus filicinus was the maximum (5.313), the minimum was Aconitum gymnandrum (0.286). The largest output was 18.000 kg x hm(-2) for Berberis agricola, the output of Saxifraga pasumensis was the lowest (0.007 kg x hm(-2)). The resources reserves of the rare and endangered plants were 15683.697 t in Lhasa region, the maximum was 7690.230 t for B. agricola, 49.03% of the total reserves, the minimum was 2.393 t for S. pasumensis, only 0.015%.

CONCLUSIONThe characteristics of rare and endangered plants were as follows: abundant species and complex habitats, widely distribution but uneven, rich reserves and high economic value. We suggested to update the endangered level of medicinal plants, strengthen the scientific research on these plants, maintain sustainable utilization of the rare and endangered plants in Lhasa region.

Base on the traditional Chinese medicinal materials list that include Pharmacopoeia of the People's Republic of China, Dao-di Herbs, the common used herbs, endangered species, to analyze the key-point investigation traditional Chinese medicinal materials list was analysed. Results displayed that the variety number of traditional Chinese medicinal materials in Pharmacopoeia of the People's Republic of China is 894, the variety number of Dao-di Herbs is 495, the variety number of the common use is 326, the variety number of traditional Chinese medicinal materials from endangered species is 280, and during the third national survey of Chinese material medica resources the variety number of traditional Chinese medicinal materials that were analysed is 360. In the list of Dao-di Herbs and common used herbs over 85% are in the list of Pharmacopoeia of the People's Republic of China, in the list of the common used herbs over 80% are in the list of Dao-di Herbs, in the list of Pharmacopoeia of the People's Republic of China and Dao-di Herbs over 10% are in the list of endangered species. The key-point investigation traditional Chinese medicinal materials list includes three part, the first part includes 563 variery traditional Chinese medicinal materials that need to statistics quantity of resource by field investigation, the second part includes 457 variery that need to monitor the changing situation, the third part includes 280 variery that need to estimated endangered situation.
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