2003 Vol. 16, No. 2

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Ozone Emitted During Copying Process -A Potential Cause of Pathological Oxidative Stress and Potential Oxidative Damage in the Bodies of Operators
2003, 16(2): 95-104.
Objective To estimate the impact of copying on the indoor air quality, and to investigatewhether ozone emitted during such a process induces pathological oxidative stress and potentialoxidative damage in the bodies of operators. Methods 67 copying operators (CO) and 67 healthyvolunteers (HV) were enrolled in a random control study, in which levels of lipoperoxide (LPO) inplasma and erythrocytes, and levels of vitamin C (VC), vitamin E (VE) and β-carotene (β-CAR) inplasma as well as activities of superoxide dismutase (SOD), catalase (CAT) and glutathioneperoxidase (GPX) in erythrocytes were determined by spectrophotometric methods. ResultsCompared with the HV group, the average values of LPO in plasma and erythrocytes in the CO groupwere significantly increased (P<0.0001), while those of VC, VE and β-CAR in plasma as well asthose of SOD, CAT and GPX in erythrocytes in the CO group were significantly decreased(P<0.0001). Pearson product-moment correlation analysis showed that with increase of ozone level incopying sites and duration of exposure to ozone, the values of LPO in plasma and erythrocytes in thebodies of operators were gradually increased,while those of VC, VE, β-CAR, SOD, CAT and GPXwere decreased in the same manner. Odds ratio (OR) of risk of biochemical parameters reflectingpotential oxidative damage of the copying operators ranged from 4.440 to 13.516, and 95 % CI of ORwas from 2.113 to 34.061. Reliability coefficient (α) of the biochemical parameters used to reflect thepotential oxidative damage of the operators was 0.8156, standardized item α=0.9929, P<0.0001.Conclusion Findings in the present study suggest that there exist a series of free radical chainreactions and pathological oxidative stress induced by high dose ozone in the operators, therebycausing potential oxidative and lipoperoxidative damages in their bodies.
A Case-control Study on Children with Guillain-barre Syndrome in North China
2003, 16(2): 105-111.
Objective To explore the risk factors for Guillain-barre syndrome. Methods Case-controlstudy design was used in 51 cases of Guillain-barre syndrome, and 51 matched controls. All of the 51cases in this study had been examined by electrophysiology. Serum IgG antibodies specific for C.jejuni were determined in all the subjects by ELISA. Each case and control were interviewed using anad hoc questionnaire, including his/her demographic information, onset of the illness, their personalhygiene and so on. Results The study showed that Guillain-barre syndrome was associated witha few factors, such as polio vaccine immunization before onset of illness (0R=7.27), no hand washingafter defecation and before meals (OR=6.15). Infection of C. jejuni was strongly associated with theillness (0R=9.5, P<0.001). Conclusion It is suggested that occurrence of Guillain-barre syndromemay correlate to infection of C. jejuni and poor personal hygiene in children.
Effects of Dietary Intervention on Hyperlipidemia in Eight Communities of Beijing, China
2003, 16(2): 112-118.
Objective To examine the effect of community-based dietary intervention on hyperlipidemia.Methods A total of 180 hyperlipidemia individuals with TG>2.26 mmol@L-1 ( 200 mg@dL-1 ) and/orTC>5.72 mmol@L-1 (220 mg@dL-1) were selected from 428 eligible subjects in eight communities ofBeijing. They were randomly divided into intervention group (n=108) and control group (n=72).Dietary intervention was provided for the intervention group for 6 months. Information on dietaryintakes, physical examinations and blood samples was collected. Serum lipids were assayed atbaseline and endpoint of the study period. Results Respective decrease in dietary intake of totalcalories, fat, cholesterol and cooking oil by 13.62%, 24.75%, 24.40%, and 22.43%, in the interventiongroup was observed. The percentages of total calories from fat, carbohydrate and protein appeared tobe desirable after study. Reduced body weight and BMI were also observed. There was a respective5.61% and 7.06 % decrease in total serum cholesterol and low-density lipoprotein cholesterol in theintervention group, while no significant changes were found in the control group. ConclusionsCommunity-based dietary intervention can effectively improve dietary patterns, control body weight,and decrease the levels of total serum cholesterol and low-density lipoprotein cholesterol.
Studies on Purification of Methamidophos Monoclonal Antibodies and Comoarative Immunoactivity of Purified Antibodies
2003, 16(2): 119-125.
Objective To purify Methamidophos (Met) monoclonal antibodies with two methods andcompare immune activity of purified antibodies. Method Caprylic acid ammonium sulphateprecipition (CAASP) method and Sepharose protein-A (SPA) affinity chromatography method wereused to purify Met monoclonal antibodies, UV spectrum scanning was used to determine proteincontent and recovery of purified antibodies, sodium dodecylsulphate polyacrylamide gelelectrophoresis (SDS-PAGE) was used to analyze the purity of purified antibodies, and enzyme-linkedimmunosorbent assay (ELISA) was used to determine immune activity of purified antibodies.Results Antibody protein content and recovery rate with CAASP method were 7.62 mg/mL and8.05% respectively, antibody protein content and recovery rate with SPA method were 6.45 mg/mLand 5.52% respectively. Purity of antibodies purified by SPA method was higher than that by CAASPmethod. The half-maximal inhibition concentration (IC50) of antibodies purified by SPA to Met was181.26 μg/mL, and the linear working range and the limit of quantification (LOD) were 2.43-3896.01μg/mL and 1.03 μg/mL, respectively. The IC50 of antibodies purified by CAASP to Met was 352.82μg/mL, and the linear working range and LOD were 10.91-11412.29 ug/mL and 3.42 μg/mL,respectively. Conclusion Antibodies purified by SPA method are better than those by CAASPmethod, and Met monoclonal antibodies purified by SPA method can be used to prepare gold-labelledtesting paper for analyzing Met residue in vegetable and drink water.
Comparison of Di-n-methyl Phthalate Biodegradation by Free and Immobilized Microbial Cells
2003, 16(2): 126-132.
Objective To compare the biodegradation of di-n-methyl pathalate by free and immobilizedmicrobial cells. Methods The enrichment and isolation technique was used to isolate themicroorganism. The PAV-entrapment method was utilized to immobilize the microorganisms. Thescanning electron microscophy (SEM) was used to observe the growth and distribution of microbialcells immobilized inside the PVA bead gels. The GC/MS method was used to identify the mainintermediates of DMP degradation. Results The microbial cells could grow quite well in PVA gel.The metabolic pathway did not change before and after immobilization of the microbial cells. Thedegradation rate of immobilized cells was higher than that of free cells. Conclusion Theimmobilized microbial cells possess advantages than free cells when applied to the biodegradation oftoxic organic pollutants.
Temperature and Daily Mortality in Shanghai: A Time-series Study
2003, 16(2): 133-139.
Objective To investigate the association between temperature and daily mortality in Shanghaifrom June 1, 2000 to December 31, 2001. Methods Time-series approach was used to estimatethe effect of temperature on daily total and cause-specific mortality. We fitted generalized additivePoisson regression using non-parametric smooth functions to control for long-term time trend, seasonand other variables. We also controlled for day of the week. Results A gently sloping V-likerelationship between total mortality and temperature was found, with an optimum temperature (e.g.temperature with lowest mortality risk) value of 26.7,℃ in Shanghai. For temperatures above theoptimum value, total mortality increased by 0.73% for each degree Celsius increase; while fortemperature below the optimum value, total mortality decreased by 1.21% for each degree Celsiusincrease. Conclusions Our findings indicate that temperature has an effect on daily mortality inShanghai, and the time-series approach is a useful tool for studying the temperature-mortalityassociation.
A Preliminary Analysis of Non-small Cell Lung Cancer Biomarkers in Serum
2003, 16(2): 140-148.
Objective To identify potential serum biomarkers that could be used to discriminate lungcancers from normal. Methods Proteomic spectra of twenty-eight serum samples from patientswith non-small cell lung cancer and twelve from normal individuals were generated by SELDI(Surfaced Enhanced Laser Desorption/Ionization) Mass Spectrometry. Anion-exchange columns wereused to fractionate the sera into 6 designated pH groups. Two different types of protein chip arrays,IMAC-Cu and WCX2, were employed. Samples were examined in PBSII Protein Chip Reader(Ciphergen Biosystem Inc) and the discriminatory profiling between cancer and normal samples wasanalyzed with Biomarker Pattern software. Results Five distinct potential lung cancer biomarkerswith higher sensitivity and specificity were found, with four common biomarkers in both IMAC-Cuand WCX2 chip; the remaining biomarker occurred only in WCX2 chip. Two biomarkers wereup-regulated while three biomarkers were down-regulated in the serum samples from patients withnon-small cell lung cancer. The sensitivities provided by the individual biomarkers were 75%-96.43%and specificities were 75%-100%. Conclusions The preliminary results suggest that serum is acapable resource for detecting specific non-small cell lung cancer biomarkers. SELDI massspectrometry is a useful tool for the detection and identification of new potential biomarker ofnon-small cell lung cancer in serum.
Purification and Immunity Analysis of Recombinant 6His- HPT Protein Expressed in E.coli
2003, 16(2): 149-156.
Objective To obtain HPT protein (Hygromycin B Phosphotransferase), a kind of plantselective maker gene product expressed from E. coli and to prepare the polyclonal antibody (pAbs)against it. Methods HPT cDNA fragment was obtained by PCR and was inserted into theprokaryotic expressing vector pBV222. Then the constructed recombinant plasmid pBV222-HPT wastransfered into E. coli DH5α for HPT expression. The recombinant expressing system was confirmedby restriction endonuclease digestion, DNA sequencing and protein expression. E. coli cells were lysedby sonication and detergent dissolution. After cell membrane was extracted, the inclusion bodies weredenatured by 8 mol/L Urea and purified with metal chelate affinity chromatography on Ni-NTAagarose under denaturing condition. The purified 6His-HPT was characterized by SDS-PAGE, andused to immunize rabbit. The titer and specificity of antisera were detected by ELISA and Westernblot respecitively. Results Analysis of DNA sequence and restricted enzymes showed that thesequence of PBV222-HPT plasmid was correct. The amount of recombinant HPT expressed in E. coliaccounted for 30% of total cellular proteins. From 1 liter of fermentative bacteria about 22 milligramsof pure recombinant HPT was isolated with purity above 95%. The recombinant HPT protein couldproduce high titer antiserum in rabbits and show good immunity activity. Western blot showedspecific binding reaction between the antiserum to the purified 6His-HPT protein and their expressedproducts (plants protein and bacterial protein). Conclusion HPT protein can be expressed andpurified from E. coli by a relatively simple method, which has high immunity activity.
Anticancer Drug Resistance of HeLa Cells Transfected With Rat Glutathione S-transferase pi Gene
2003, 16(2): 157-162.
Objective To establish a cytologic expressing system of rat glutathione S-transferase pi(GST-pi) cDNA for detecting the resistance of HeLa cells to anticancer drugs. Methods Theassessment was made with various anticancer drugs (adriamycin, mitomycin, cisplatinum andvincristine) that showed different cytotoxicities in transfectant HeLa cells with pSV-GT containing ratGST-pi cDNA (HeLa/pSV-GT) or control pSV-neo (HeLa/pSV-neo). Expression levels of GST-pimRNA in HeLa/pSV-GT and HeLa/pSV-neo were measured by in situ hybridization usingDigoxin-labelled cDNA probe. Results HeLa/pSV-GT expressed significantly high degree ofGST-pi mRNA, whereas both HeLa/pSV-neo and HeLa cells had very low expression. Cytotoxicitiesof HeLa/pSV-GT and HeLa/pSV-neo with 4 anticancer drugs were measured by MTT assay. Drugconcentrations for yielding 50% inhibition (IC50) in HeLa/pSV-GT by adriamycin, mitomycin andcisplatinum were 70.13 μg/mL, 10.95 μg/mL and 16.52 μg/mL, respectively. In contrast, IC50 inHeLa/pSV-neo was 10.34 μg/mL, 7.48 μg/mL and 13.70 μg/mL, respectively. The cytotoxicities ofvincristine on both HeLa/pSV-GT and HeLa/pSV-neo were not significantly different. ConclusionsOur findings suggest that HeLa/pSV-GT containing rat GST-pi cDNA is resistant to some anticancerdrugs due to overexpression of GST-pi. Also, HeLa/pSV-GT cell line could serve as a usefulcytogenetic model for further research.
Characterization of Phenol Biodegradation by Comamonas testosteroni ZD4-1 and Pseudomonas aeruginosa ZD4-3
2003, 16(2): 163-172.
Objective To investigate the characteristic and biochemical mechanism about the phenolbiodegradation by bacterial strains ZD 4-1 and ZD 4-3. Methods Bacterial strains ZD 4-1 and ZD4-3 were isolated by using phenol as the sole source of carbon and energy, and identified by 16SrDNA sequence analysis. The concentrations of phenol and total organic carbon (TOC) weremonitored to explore the degradation mechanism. The biodegradation intermediates were scanned at375 nm by using a uv-vis spectrophotometer. The enzyme assays were performed to detect theactivities of dioxygenases. Results Bacterial strains ZD 4-1 and ZD 4-3 were identified asComamonas testosteroni and Pseudomonas aeruginosa by 16S rDNA sequence analysis, respectively.The growth of the two strains was observed on a variety of aromatic hydrocarbons. The strains ZD 4-1and ZD 4-3 metabolized phenol via ortho-pathways and meta-pathways, respectively. In addition, theresults of enzyme assays showed that the biodegradation efficiency of phenol by meta-pathways washigher than that by ortho-pathways. Finally, the results of induction experiment indicated that thecatechol dioxygenases, both catechol 1,2-dioxygenase (C120) and catechol 2,3-dioxygenase (C230),were all inducible. Conclusion The strains ZD 4-1 and ZD 4-3 metabolize phenol throughortho-pathways and meta-pathway, respectively. Furthermore, the biodegradation efficiency of phenolby meta-pathways is higher than that by ortho-pathways.
Parkinson's Disease and Smoking: An Integral Part of PD's Etiological Study
2003, 16(2): 173-179.
Objective To explore the association of Parkinson's disease (PD) with cigarette smoking.Methods One hundred of fourteen PD patients were compared with 205 control subjects who werematched by gender, race and residency. A previously validated questionnaire including smoking,alcohol/tea consumption as well as some other environmental exposure data was administered.Results With never-smokers as the reference category, we observed reduced risk for PD among eversmokers (OR=0.49, 95% CI: 0.30 to 0.79) current smokers (OR=0.44, 95% Cf: 0.23 to 0.86) andex-smokers (OR=0.54, 95% Cf: 0.30 to 0.96). When ever smokers were stratified by years of smoking,there was an inverse correlation between those whose smoking history was longer than 20 years(OR=0.40 95% CI: 0.21 to 0.81) and an even mild protective correlation between those who smokedless than 20 years (OR=0.57, 95% CI: 0.33 to 0.99). Those who had quitted smoking for more than 20years were less likely to have the disease than never smokers, and those who had quitted for less than20 years were least likely to have PD, while those who were current smokers were still least likely tohave the disease. We found significant inverse gradient with pack-day smoking (trend P<0.05), andthe inverse correlation between cigarette smoking and PD was not confounded by alcohol/teaconsumption and other confounding bias. Conclusions The inverse correlation between Parkinson'sdisease risk and smoking as well as the trend of gradient dose response is again observed in our study.More future researches are needed to confirm these correlations and to explore further biochemicalevidence.
A Survey on Injury Incidence in School Children in Shantou City, China
2003, 16(2): 180-186.
Objectives To study incidence characteristics and causes of injury, and its medicalconsequences in school children of China. Methods A total of 2 553 school children aged 7-16years were recruited from Shantou City in Gunagdong by cluster sampling method, and wereinvestigated with questionnaires on cases of injuries occurred among them from October 1, 1996 toSeptember 30, 1997. Results Injuries tended to increase with children's age, with an overallincidence rate of 37.96%, higher in boys than in girls (P<0.05); and 38.1% of children had more thantwo episodes of injury during this period. Falls took leading place of injury incidence both in boys andgirls and in all age groups. Most injuries occurred when they were playing, sporting, riding andwalking at home or in school. Self-inflicted injury ranked the first place of all injuries, followed byhurt caused by others (classmates, sibling or others). Moderate and serious injuries accounted for 8%of the total with a disability rate of 121.4/100 000. Conclusions Currently, injury has become oneof the serious public health problems in China. For the improvement of children survival, it is crucialto reduce their injury to strengthen research on child safety and to implement safety-promotionprograms.
Effects of Malnutrition on Child Survival in China As Estimated by PROFILES
2003, 16(2): 187-193.
Objective To estimate the benefits of reductions in underweight and Vitamin A deficiency forchild survival in China that might be expected as a result of lowering the prevalence of theseconditions. Methods Profiles, a process of nutrition policy analysis was used to quantify thefunctional consequences of malnutrition in terms of child survival. Results Underweight Theactual reduction in underweight between 1992 and 2001 (from 15.7% to the current 10.1%) resultedin saving of 176 000 child lives. As estimated, without improvements, 612 000 children will die due tounderweight between 2001 and 2010, 281 000 (46%) of them living in western provinces. Reducingunderweight prevalence from 10.1% to 8% could overall save 62 000 lives. The reduction ofunderweight prevalence in the west alone might save 56 000 lives. Vitamin A in China as a whole,vitamin A deficiency accounts, as estimated, for 7.5% of deaths of children 6-59 months old,representing 206 000 deaths over the past ten years. Halving the prevalence over the period wouldsave 49 000 child lives. The higher prevalence and higher mortality rates in western provinces meanthat even with only 28% of the Chinese population, over half of child deaths there are related tovitamin A.