2008 Vol. 21, No. 1
2008, 21(1): 1-6.
Objective To explore the effects of nonylphenol on brain gene expression profiles in F1 generation rats by microarray technique.Methods mRNA was extracted from the brain of 2-day old F1 generation male rats Whose F0 female generation was either exposed to nonylphenol or free from nonylphenol exposure,and then it was reversely transcribed to cDNA hbeled with cy5 and cy3 fluorescence.Subsequently,cDNA probes were hybridized to two BiostarR-40S cDNA gene chips and fluorescent signals of cy5 and cy3 were scanned and analyzed. Results Two genes were differentially down-regulated.Conclusion Nonylphenol may disturb the neurcendocrine function of male rats when administered perinatally.
2008, 21(1): 7-23.
Objective BacoMindTM (BM) is a standardized extract of Bacopa monnieri,which belongs to the family Scrophulariaceae and is a creeping annual plant found throughout the Indian subcontinent.It has been used by Ayurvedic medicinal practitioners in India for almost 3000 years and is classified as a medharasayana,a substance which improves memory and intellect.With the widespread traditional use as well as scientific validation of Bacopa monnieri for nootropic activity,a bioactive-rich unique Phvtochemical composition-BacoMindTM was developed from B.monnieri for use as a cognition and memory enhancing agent. The present study aimed to investigate the in vitro toxicity of this formulation of BacoMindTM on human lymphocytes and to rule out its possible contribution to mutagenicity. Methods In the present investigation the active ingredients present in BM were identified and quantified by high performance liquid chromatography (HPLC)and high performance thin-layer chromatography(HPTLC).Antioxidant and anticlastogenic properties of BM were studied in vitro with and without metabolic activation.Doses of BM were chosen on the basis of mitotic index(MI)and cytokinesis-block proliferation index(CBPI).Clastogenicity assays were performed at 31.2 μg/mL,62.5 μg/mL,and 125 μg/mL,while the Salmonella reverse mutation assay (Ames test)was performed at doses of 61.72,185.18,555.55,1666.67,and 5000.00 μg/plate. Results HPLC and HPTLC analysis of BM revealed the presence of bacosidc A3,bacopaside I,bacopaside Ⅱ,iujubogenin isomer of bacopasaponin C,bacosine,luteolin,apigemn,bacosine,and β-sitosterol D glucoside.BM demonstrated significant antioxidant activity.The number of chromosomal aberrations and the frequency of micronuclei induced by BM were not statistically significant up to a dose of 62.5 μg/mL.A subsequent dose of 125 μg/mL prior to metabolic activation induced mild clastogenicity,but it was found to be biologically insignificant as this effect was not seen post metabolic activation.BM also demonstrated a dose-dependent protection against the clasmgens used in this study using the above tests for clastogenicity.Maxilnum protection was observed in presence of metabolic activation.Moreover,BM demonstrated no mutagenic effect on the tested strains,as observed in the Ames test.Conclusion BM protected human lymphoeytes against various clastogens.BM also exhibited high antioxidant activity which might be responsible for the observed protective effects against the clastogens since the used clastogens are known to indace their clastogenic effects via production of oxidative radicals.
Screening of Antioxidant Potential of Selected Barks of Indian Medicinal Plants by Multiple in vitro Assays LI
2008, 21(1): 24-29.
Objective To evaluate the antioxidant potential in herbal extract barks of five therapeutically important medicinal plants native to India,i.e.Crataeva nurvala Buch.-Ham.,Buchanania lanzan Spreng.,Aegle marmelos Corr.,Dalbergia sissoo Roxb.ex DC.,and Cedrela toona Roxb.Methods Standardized aqueous alcoholic extracts from the selected barks having different target radicals,such as superoxide radical,nitric oxide,ABTS radical,and peroxidative decomposition of phosphohpids.were prepared and screened bv multiple in vitro assays.These extracts were also tested for total phenolic and tannin content and correlated with antioxidant capacity. Results Tbtal phenolic and tannin contents were found to be the highest in C. nurvala (195 GAE mg/g and 218.3 mg/g CE).SOD mimetic activity was found to be the highest in Crataeva nurvula,although all barks showed activity more than 100 units/mg extract.Lipid peroxidation inhibitory potential was found to be the highest in Crataeva nurvala(83.4% inhibition of MDA formation/10 μg extract),and also showed a comparatively high NO quenching capacity (45.5% per 10 μg extract).The highest NO quenching potential was found in Aegle marmelos(47.3% per 10 μg extract).Cedrela toona showed the lowest LPO inhibitory potential and NO quenching capacity(50.5% and 30.5%,respectively).Buchanania lanzan,a medicinal plant extensively used for inflammatory disorders and Dalbergia sissoo also showed 72.5% and 69.1% LPO inhibitory potential/10 μg extract.Trolox equivalent antioxidant capacity ranged from 0.24 to 0.39 mmol/L TEAC/mg extract,indicating that all the barks tested had ABTS+ radical quenching capacity.Conclusion Bark of Crataeva nurvulahas the highest antioxidant capacity and a positive correlation between antioxidant activity and their plendic content was found.
Different Patterns of Cyclin D1/CDK4-E2F-1/4 Pathways in Human Embryo Lung Fibroblasts Treated by Benzo[a]pyrene at Different Doses1
2008, 21(1): 30-36.
Objective To investigate the roles of the cyclin D1/CDK4 and E2F-1/4 pathways and compare their work patterns in cell cycle changes induced by different doses of B[a]P. Methods Human embryo lung fibroblasts(HELFs)were treated with 2 μmol/L or 100 μmol/L B[a]P which were provided with some characteristics of transformed cells (T-HELFs).Cyclin D1,CDK4 and E2F-1/4 expressions were determined by Westem blotting.Flow cytometry was used to detect the distribution of cell cycle.Results After B[a]P treatment,the proportion of the first gap(G1)phase cells decreased.CDK4 and E2F-4 expression did not change significantly.In 2 μmol/L treated cells,a marked overexpression of cyclin D1 and E2F-1 was observed.However,in T-HELFs overexpression was limited to cyclin D1 only,and no overexpression of E2F-1 was observed.The decreases of G1 phase in response to B[a]P treatment were blocked in antisense cyclin D1 and antisense CDK4 transfected HELFs (A-D1 and A-K4)and T-HELFs(T-A-D1 and T-A-K4).Atier 2 μmol/L B[a]P treatment,overexpression of E2F-1 was attenuated in A-D1,and E2F-4 expression was decreased significantly in A-K4.In T-A-D1 and T-A-K4,E2F-4 expression was increased significantly,compared with T-HELFs.The E2F-1 expression remained unchanged in T-A-D1 and T-A-K4.Conclusions Cyclin D1/CDK4-E2F-1/4 pathways work in different patterns in response to low dose and high dose B[a]P treatment.In HELFs treated with 2 μmol/L B[a]P, cyclin D1 positively regulates the E2F-1 expression while CDK4 negatively regulates the E2F-4 expression;however,in HELFs treated with 100 μmol/L B[a]P,both cyclin D1 and CDK4 negatively regulate the E2F-4 expression.
2008, 21(1): 37-44.
Objective To investigate the association between birth defects and dietary nutrient intake in a high risk area of China.Methods A dietary survey was performed and serum folic acid was measured in women whose pregnancy was affected by neural tube defects(NTDs)or unaffected by any birth defects(BDs)in Zhongyang and Jiaokou Counties in Shanxi Province of China.Results The local average censureption of foods including dark green vegetables,fluits,fat and meat,and nutrient intake(e.g.energy,protein,retinol,riboflavin,vitamin E,and selenium)were lower than the national average level.In women of childbearing age,these regions,the intake of nutrients was much lower than the recommended nutrient intake(9%-77%)The case-centrel dietary nutrition smdv of women whose pregnancy was affected bv BDs(including MTDs and congenital heart defects)demonstrated that,in early pregnancy,adequate nutrition(I.e.eating meat,fresh vegetables,fruit more than once a week)was a protective factor,while eating germinated potatoes was a risk factor.The geometrical mean(p5-p95)of serum folic acid in women with NTD birth defects was 9.6 nmol/L(3.6,23.03),which was significantly lower than that in normal women(14.03 nmol/L). Conclusion Wemen of childbearing age in the two counties of Shanxi Province,Chim,have a marked insufficient intake of some nutrients,especially folic acid,zinc,vitamins A and B12.This nutrient deficiency may be an important risk factor for the high prevalence of birth defects in these regions.Therefore,adequate dietary nutrition in early pregnancy can prevent BDs.
Using Support Vector Machine to Predict Eco-environment Burden:A Case Study of Wuhan,Hubei Province,China1
2008, 21(1): 45-52.
Objective The human socio-economic development depends on the planet's natural capital.Humans have had a considerable impact on the earth,such as resources depression and environment deterioration.The objective of this study was to assess the impact of socio-eeonomic development on the ecological environment of Wuhan,Hubei Province,China,during the general planning period 2006-2020. Methods Support vector machine(SVM) model was constructed to simulate the process of eco-economic system of Wuhan.Socio-economic factors of urban total ecological footprint(TEF)were selected by partial least squares(PLS)and leave-one-out cross validation(LOOCV).Historical data of socio-economic factors as inputs,and corresponding historical data of TEF as target outputs.were presented to identify and validate the SVM model.When simulated as output in succession. Results Up to 2020,the district would have suffered an accumulative TEF of 28.374 million gha,which was over 1.5 times that of 2004 and healrly 3 times that of 1988.The per capita EF would be up to 3.019 gha in 2020. Conclusions The simulation indicated that although the increase rate of GDP would be restricted in a lower level during the general planning period,urban ecological environment burden could not respond to the socio-economic circumstances promptly.SVM provides tools for dynamic assessment of regional eco-environment.However,there still exist limitations and disadvantages in the model.We believe that the next logical step in deriving better dynamic models of ecosystem is to integrate SVM and other algorithms or technologies.
Detection of Genetically Modified Crops by Combination of Multiplex PCR and Low-density DNA Microarray1
2008, 21(1): 53-62.
Objective To develop a technique for simultaneous detection of various target genes in Roundup Ready soybean by combining multiplex PCR and low-density DNA microarray.Methods Two sets of the multiplex PCR system were used to amplify the target genes in genetically modified(GM)soybean.Seventeen capture probes(PCR products)and 17 pairs of corresponding primers were designed according to the genetic characteristies of Rroundup Ready soybean(GTS40-3-2),maize (Mort810,Nk603,GA21),canola(T45,MS1/RF1),and rice(SCK)in many identified GM crops.All of the probes were categorized and identified as species-specific probes.One negative probe and one positive control probe were uscd to assess the efficiency of all reactions,and therefore eliminate any false positive and negative results.After multiplex PCR reaction,amplicons were adulterated with Cy5-dUTP and hvbridized with DNA microarray.The array was then scanned to display the specific hybridization signals of target genes.The assay was applied to the analysis of sample of ccrtified transgenic soybean (Roundup Ready GTS40-3-2)and canola(MS1/RF1). Results A combination technique of multiplex PCR and DNA microarray was successfully developed to identify multi-target genes in Roundup Ready soybean and MS1/RF1 canola with a great specificity and reliability.Reliable identification of genetic characteristics of Roundup Ready of GM soybean from genetically modified crops was achieved at 0.5% transgenic events,indicating a high sensitivity. Conclusion A combination technique of multiplex PCR and low-density DNA microarray can reliably detect and identify the genetically modified crops.
2008, 21(1): 63-68.
Objective To investigate the immunological effect of PM2.5 on cytokine production in female Wistar rats.Methods Female Wistzr rats were given 0.3 mg,0.75 mg,2 mg,5 mg of PM2.5 per 0.5mL saline,respectively.Saline was used as the negative control.TNF-α and IL-6 levels in the branchoalveolar lavage were measured by ELISA,and mRNA expression leveIs in lung tissue were detected bv RT-PCR.Alveolar macrophages were collected for testing phogacytic function. Results Exposure to PM2.5 stimulated TNF-α production in a dose-dependent manner(P＜0.05),However,no statistically significant difference was found.No time-dependent change in TNF-α and IL-6 production Was found.TNF-α and IL-6 mRNA expressions were induced by PM2.5-exposure.The phagocytic rate(PR)was significantly decreased by PM2.5 treatment.Conclusion PM2.5 exposure increases inflammation response of the lung in a dose-dependent mauuer.Moreover,tissue injury induced by PM2.5 may be related to altered production of cytokines.PM2.5 may impair the phagocytic activity of alveolar macrophages.
Establishment of a Sandwich ELISA Method for Detection of Vascular Endothelial Growth Factor in Serum Samples of Hepatocellular Carcinoma Patients1
2008, 21(1): 69-74.
Objective To establish a sandwich ELISA method for detecting vascular endothelial growth factor(VEGF)in sera of population and the patients with hepatocellular carcinoma(HCC). Methods Full length and two truncated human VEGF cDNA sequences were amplified from a commercial plasmid pBLAST49-bVEGF by PCR and inserted into the prokaryotic-expression plasmid pET-32a or pGEX-2T. Various VEGF proteins were expressed and purified from E. coli in His-Trx or GST fusion forms.The specific VEGF antibodies were elicited in experimental rabbits and mice by immunization of the full length VEGF fusion protein His-Trx-VEGF1-165.After purification of antibodies with chromatograph of Protein G.a sandwich ELISA technique was established.Serum VEGF levels were evaluated in 229 adults and 291 HCC patients.Results SDS-PAGE displayed that the molecular weights of the expressed full length(His-Trx-VEGF1-165),N-terminal(His-Trx-VEGF1-100)and C-terminal(GST-VEGF100-165)human VEGF fusion proteins were about 38KD,31KD,and 33KD,respectively.Western blots confirmed that the prepared antisera were able to recognize both prokaryoticly and eukaryoticly expressed recombinant VEGF proteins.Assays of serially diluted His-Trx-VEGF1-100 by the established sandwich ELISA method showed that the linear range of the standard curve was 0.625-320 ng/mL.with the squared correlation coefficient R2=0.991.Screening of a serum panel containing 291 serum samples of HCC patients and 229 health adults rovealed that the average VEGF level in HCC patients was higher than that in healthy controls,with a statically significant difference. Conclusion The established sandwich ELISA reflects the level of serum VEGF and provide scientific basis for scrcening metastasis and recurrence of HCC using serum VEGF as an index.
Evaluation of Effectiveness of Integrated Intervention Program in Improving Drug Addicts' Psychological Health1
2008, 21(1): 75-84.
Objective To investigate the social mental state of drug addicts in a compulsive drug abuse treatment center;evaluate the effectiveness of integrated program for the prevention of abuse relapse and improvement of drug addicts' psychological health.Methods The study subjects were addicts from the Wuhan Compulsive Drug Abuse Treatment Center between October 2003 and June 2004,who satisfied the inclusion criteria.A non-randomized control-intervention study design was adopted.Volunteers willing to take part in intervention were put into the intervention group with their full awareness and willingness to prevent drug abuse relapse.The control group was composed of the addicts who were willing to prevent relapse and to be followed up after their discharge. Results The effectiveness of the integrated intervention program in promoting addicts' psychological health:before the intervention,the scores of Self-Rating Anxiety Scale(SAS),the positive and negative dimensionalities of Simple Coping Style Questionnaire(SCSQ)and Chinese Perceived Stress Scales(CPSS)had no significant differences between the intervention group and the control group.After the intervention,exccpt that the SCSQ's positive dimensionality in the intervention group was significantly higher than that in the control group,other indices in the intervention group were lower.Before and after the intervention.the psychological health level in both the groups was lower than that in the normal population;there were significant differences between addicts and normal subjects in regards with all of the indices above. Conclusion Drug abuse was associated closely with addicts' social mental factors.The integrated intervention program can alleviate anxiety and stress,reduce co-morbid mental disorders and effectively improve their coping style.In conclusion,the program can promote addicts' psychological health significantly.
2008, 21(1): 85-89.
Objective To verify whether Warthin-Starry(WS)silver method could detect the air particulate matter(PM)/dust particles(Ps)located within the macrophages in situ. Methods There were 26 antopsy cases that resulted from cerebral hemorrhage(group A),silicosis(group B),and fetal death during pregnancy(group C).Samples were collected separately and serial sections were prepared from the lungs and lymph nodes and stained with hematoxylin and eosin(HE),WS silver,immunohistochemistry of CD68.Furthermore,ultrathin sections were taken from the WS positive serial sections of groups A and B.Ps were observed under a transmission electron microscope(TEM)and the elements of Ps were measured by X-ray spectrum analysis(X-RSA).Results In both groups A and B,WS staining was positive for the larger and fine Ps,the so called"dust cells",but HE staining Was almost negative for fine Ps.In group C,no larger or fine Ps were found.Immunohistochemical staining of CD68 certified that the"dust cells"containing Ps were macrophages.The results of TEM and X-RSA proved that the structure and elements of Ps belonged to PM indeed.Conclusion WS staining is a better than HE staining in showing the location of PM within macrophages.