2007 Vol. 20, No. 3
Significant Positive Correlation of Plasma BPDE-Albumin Adducts to Urinary 1-Hydroxypyrene in Coke Oven Workers
2007, 20(3): 179-183.
Objective To investigate the application of BPDE-albumin adducts as monitoring biomarkers for coke oven workers exposed to polycyclic aromatic hydrocarbons(PAHs)and to explore possible relationship between BPDE-albumin adducts and urinary 1-hydroxypyrene (1-OHP)levels in them.Methods Thirty-seven coke oven workers from a coke plant and 47 controls without the occupational exposure to PAHs were recruited in this study.The levels of plasma BPDE-albumin adducts and urinary 1-OHP were analyzed using high performance liquid chromatography.Results The median levels of BPDE-albumin adducts(42.10 fmol/mg albumin)and urinary 1-OHP(5.46 μmol/mol creatinine)were significantly higher in coke oven workers than in controls(14.16 fmol/mg albumin,2.96 μmol/mol creatinine,respectively;P＜0.01).Multiple logistic regression analysis showed that coke oven workers were at higher risk of having BPDE-albumin adduct levels above 25.30 prnol/mg albumin(OR＝1.79,P＜0.01)and urinary 1-OHP levels above 4.13 μmol/mol creatinine(OR=2.45,P＜0.05).There was a positive correlation between the levels of BPDE-albumin adducts and urinary 1-OHP in all subjects(rs＝0.349,P＜0.01).Conclusion BPDE-albumin adduct is a useful biomarker for monitoring long-term exposure to PAHs,and plasma BPDE-albumin adducts level is significantly correlated to urinary 1-OHP levels in coke oven workers.
2007, 20(3): 184-188.
Objective To produce specific monoclonal antibody(mAb)against recombinant human erythropoietin(rHuEPO)for development of higmy efficient methods for erythropoietin detection in biological fluids.Methods rHuEPO was covalently coupled with bovine serum albumin(BSA)and the conjugate was used to immunize mice to produce specific mAb against rHuEPO based on hybridoma technology.The obtained F3-mAb was characterized by enzyme-linked immunosorbent assay (ELISA),SDS-PAGE and Western blot.Results The isotype of F3-mAb Was found to be IgM with an affinity constant of 2.1x108 L/mol.The competitive ELISA using the obtained IgM showed a broader linear range and lower detection limit compared with previous work.Conclusions The modification of rHuEPO was proved to be successful in generating required specific mAb with high avidity to rHuEPO.
2007, 20(3): 189-197.
Objective To investigate the effect of lidamycin(LDM)on telomerase activity in human hepatoma BEL-7402 cells under the condition of LDM inducing mitotic cell death and senescence.Methods Chromatin condensation was detected by co-staining with Hoechst 33342 and PI.Cell multinucleation was observed by Giemsa staining and genomic DNA was separated by agarose gel electrophoresis.Fluorescent intensity of Rho123 Was determined for mitochondrial membrane potential.MTT assay and SA-β-gal staining were employed to analyze the senescence-like phenotype.The expression of proteins was analyzed by Western blot.Telomerase activity was assayed by telomerase PCR-ELISA.Results Mitotic cell death occurred in LDM-treated cells characterized by unique and atypical chromatin condensation,multinucleation and increased mitochondrial membrane potential.However,no apoptotic bodies or DNA ladders were found.In addition,apoptosis-related proteins remained nearly unaltered.Senescence-like phenotype was identified by increased and elongated size of cells,growth retardation,enhanced SA-β-gal activity and the changes of senescence-related protein expression.Telomerase activity markedly decreased (P＜0.01)in LDM-treated hepatoma BEL-7402 cells. Conelusion Mitotic cell death and senescence could be triggered simultaneously or sequentially after exposure of hepatoma BEL-7402 cells to LDM.The decrease in telomerase activity may play a key role in the defective mitosis and aging morphology.Further investigation of detailed mechanism is needed.
Treatment of Scrapie Pathogen 263K With Tetracycline Partially Abolishes Protease-resistant Activity in vitro and Reduces Infectivity in vivo
2007, 20(3): 198-202.
Objective To study the possible effect of tetracycline on protease-resistant activity in vitro and infectivity in vivo of a scrapie strain 263K.Methods Scrapie pathogens were incubated with tetracycline at different concentrations for various periods of time and protease-resistant PrP signals were evaluated with proteinase K-treatment and Western blots.The preparations treated with tetracycline were intracerebrally inoculated into golden hamsters and typical TSE manifestations were noted.PrPSc in brain tissues of the infected animals was detected by PrP specific Western blot assays. Results Protease-resistant PrP was significantly reduced in or removed from the preparations treated with tetracycline in a dose-dependant manner.Compared with the control group after incubated for 53.75±0.50 days,the preparations treated with 5 mmol/L and 20 mmol/L tetracycline prolonged the incubation time of 61.5±1.73 and 59.5±0.58 days (P＜0.05). Conclusion Treatment of scrapie pathogen 263K with tetracycline reduces or removes its protease-resistant activity in vitro.
2007, 20(3): 203-207.
Objective To determine the metal contents of lichen species from East Black Sea region of Turkey for investigation of trace metal pollution sourced traffic.Methods The levels of copper,cadmium,lead,zinc,manganese,iron,chromium,nickel,cobalt,palladium in lichen samples collected from East Black Sea region of Turkey were determined by flame and graphite furnace atomic absorption spectrometry after microwave digestion method.The accuracy of the method was corrected by standard reference material(NIST SRM IAEA-336 Lichen). Results The contents of investigated trace metals in lichen samples were 7.19-22.4 μg/g for copper,0.10-0.64 μg/g for cadmium,4.03-44.6 μg/g for lead,14.5-41.8 1.μg/g for zinc,25.8-208 μg/g for manganese,331-436 μg/g for iron,1.20-3.01 μg/g for chromium,1.48-3.90 μg/g for nickel,0.20-3.55 μg/g for cobalt.0.11-0.64 μg/g for palladium.The results were compared with the literature values.Conclusion Some lichen species such as Xanthoparmelia conspersa,Xanthoria calcicola,Peltigera membranacea,and Physcia adscendens are accumulated trace metals at a high ratio.
Conceptual Model for Automatic Early Warning Information System of Infectious Diseases Based on Internet Reporting Surveillance System
2007, 20(3): 208-211.
Objective To establish a conceptual model of automatic early warning of infectious diseases based on internet reporting surveillance system,with a view to realizing an automated warning system on a daily basis and timely identifying potential outbreaks of infectious diseases. Methods The statistic conceptual model was established using historic surveillance data with movable percentile method.Results Based on the infectious disease surveillance information platform,the conceptualmodelfor early warning was established.The parameter,threshold,and revised sensitivity and specificity of early warning value were changed to realize dynamic alert of infectious diseases on a daily basis.Conclusion The instructive conceptual model of dynamic alert can be used as a validating tool in institutions of infectious disease surveillance in different districts.
2007, 20(3): 212-216.
Objective To study the membolism of 14C-deltamethrin in rats. Methods Rats were dosed orally and i.p.with a single dose of 14C-deltamethrin(0.64 mg/Kg)body weight.The required dose was applied daily for 3 days.At the end of the experiment,selected organs,such as liver,kidney,fat,intestine,and blood were excised for radioassay of 14C-content.Results Deltamethrin was almost eliminated from the body within 1-3 days.The main portion of 14C-residues Was extracted from urine(38%,32%)and feces(20%,24%)with a little amount remained in various organs. Conclusion The elimination and distribution of 14C-radioactivity in rats treated orally and intraperitoneally signify th6at deltamethrin is bioavailable in urine and feces.
2007, 20(3): 217-225.
Objective To investigate the feasibility of reducing THM precursors and controlling bromate taste and odor in drinking water taken from the Yellow River by an ozonation combined system. Methods The appropriate ozone dosage was determined,and then the changes of TOC,UV254 and THM formation potential(THMFP)in the combined system were evaluated. Results One mg/L ozone could effectively remove taste and odor and meet the maximum allowable bromate level in drinking water.The pre-ozonation increased THMFP,but the conventional treatment system could effectively reduce the odor.The bio-ceramic filter could partly reduce CHCI3FP,but sometimes might increase CHCI2BrFP and CHCIBr2FP.The biological activated carbon(BAC)filter could effectively reduce CHCI3FP and CHCI2BrFP,but increase CHCIBr2FP.Compared with other filters.the fresh activated carbon(FAC)filter performed better in reducing THMFP and even reduced CHCIBrzFP.Conclusion The combined system can effectively reduce taste,odor,CHCI3FP,and CHCI2BrFP and also bring bromate under control.
Modulatory Effect of Distillate of Ocimum sanctum Leaf Extract(Tulsi)on Human Lymphocytes Against Genotoxicants
2007, 20(3): 226-234.
Objective To study the modulatory effect of distillate of Ocimum sanctum(traditionally known as Tulsi)leaf extract (DTLE)on genotoxicants.Methods In the present investigation,we studied the antigenotoxic and anticlastogenic effect of distillate of Tlulsi leaf extract on(i)human polymorphonuclear leukocytes by evaluating the DNA strand break without metabolic activation against mitomycin C(MMC)and hexavalent chromium(Cr+6)and(ii)human peripheral lymphocytes (in vitro)with or without metabolic activation against mitomycin C(MMC),hexavalent chromium(Cr+6)and B[a]P by evaluating chromosomal aberration(CA)and micronucleus assay(MN).Three different doses of DTLE,50 μL/mL,100 μL/mL,and 200 μL/mL were selected on the basis of cytotoxicity assay and used for studying DNA strand break,chromosomal aberration and micronucleus emergence.The following positive controls were used for inducing genotoxicity and clastogenicity:MMC(0.29 μmol/L)for DNA strand break,chromosomal aberration and 0.51 μmol/L for micronucleus assay;Potassium dichromate(Cr+6)600 μmol/L for DNA strand break and 5 μmol/L for chromosomal aberration and micronucleus assay;Benzo[a]pyrene(30 μmol/L)for chromosomal aberration and 40 μmol/L for micronucleus assay.The active ingredients present in the distillate of Tulsi leaf extract were identified by HPLC and LC-MS.Results Mitomycin C(MMC)and hexavalent chromium(Cr+6) induced statistically significant DNA strand break of respectively 69%and 71%(P＜0.001)as revealed by fluorometric analysis of DNA unwinding.Furthermore,thedamage could be protected with DTLE(50 μL/mL,100 μL/mL,and 200 μL/mL) on simultaneous treatment.Chromosomal aberration and micronucleus formation induced by MMC,Cr+6 and B[a]P were significantly protected(P＜0.001)by DTLE with and without metabolic activation.Conclusion Distillate of Tulsi leaf extract possesses antioxidants contributed mainly by eugenol,luteolin and apigenin as identified by LC-MS.These active ingredients may have the protective effect against genotoxicants.
2007, 20(3): 235-241.
Objective To study the transfer of paralytic shellfish toxins(PST)using four simulated marine food chains:d8noflagellate Alexandrium tamarense→Artemia Anemia salina→Mysid shrimp Neomysis awatschensis;A.tamarense→N.awatschensis:A.tamarense→A.salina→Perch Lateolabrax japonicus;and A.tamarenae→L. japonicus. Methods The ingestion of A.tamarenge.a producer of PST, by L. japonicus,N.awatschensis,and A.salina was first confirmed by microscopic observation of A.tamarense cells in the intestine samples of the three different organisms,and by the analysis of Chl.a levels in the samples.Toxin accumulation in L.japonicus and N.awatschensis directly from the feeding on A.tamarense 8r indirectly through the vector of A.salina was then studied.The toxicity of samples was measured using the AOAC mouse bioassay method,and the toxin content and profile of A.tamarense were analyzed by the HPLC method.Results Both A.salina and N.nwatschensis could ingest A.tamarense cells.However,the ingestion capability of A.salina exceeded that of N.L. japonicus.N.awatschensis and A.salina by microscopic observation.Therefore,the three organisms could ingest A.tamarense cells directly.A.salina could accumulate high content of PST,and the toxicity of A.salina in samples collected on of tested mice within 7 minutes,and the toxin content in artemia sample collected on the lst day was estimated to be 1.65×10-5 μg STX equal/individual.Toxin accumulation in L.japonicus and N.awatschensis directly from the feeding on A.tamarense or indirectly from the vector of A.salina was also studied.The mice injected with extracts from L.japonicus and N. awatschensis samples that accumulated PST either directly or indirectly showed PST intoxication symptoms,indicating that low levels of PST existed in these samples Conclusion Paralytic shellfish toxins can be transferred to L.japonicus,N.awatschensis,and A.salina from A.tamarense directly or indirectly via the food chains.
Induction of Functional Recovery by Co-transplantation of Neural Stem Cells and Schwann Cells in a Rat Spinal Cord Contusion Injury Model
2007, 20(3): 242-249.
Objeetive To smdy the transplantation efficacy of neural stem cells(NSCs)and Schwann cells(SC)in a rat model of spinal cord contusion injury.Methods Multipotent neural stem cells(NSCs)and Schwann cells were harvested from the spinal cords of embryomc rats at 16 days post coitus and sciatic nerves of newborn rats,respectively.The differential characteristics of NSCs in vitro induced by either serunl-based culture or co-culture with SC were analyzed by immunofluorescence.NSCs and SCs were co-transplanted into adult rats having undergone spinal cord contusion at T9 level.The animals were weekly monitored using the Basso-Beattie-Bresnahan locomotor rating system to evaluate functional recovery from contusion-induced spinal cord injury.Migration and differentiation of transplanted NSCs were studied in tissue sections using immunohistochemical staining.Results Embryomc spinal cord-derived NSCs differentiated into a large number of oligodendrocytes in serum-based culture upon the withdrawal of mitogens.In cocultures with SCs,NSCs differentiated into neuron more readily.Rats with spinal cord contusion injury which had undergone transplantation of NSCs and SCs into the intraspinal cavity demonstrated a moderate improvement in momr functions.Conclusions SC may contribute to neuronal differentiation of NSCs in vitro and in vivo.Transplantation of NSCs and SCs into the affected area may be a feasible approach to promoting motor recovery in patients after spinal cordin jury.
2007, 20(3): 250-259.
Objective To evaluate the protective efficacy of gossypin(3,3',4',5,7,8-hexahydroxyflavone 8-glucoside)by administering it intraperitoneally,for dose,time,and vehicle dependent effects against sulphur mustard(SM),administered through percutaneous route in mice.Methods SM(diluted in PEG-300)was administered percutaneously.The protective efficacy of gossypin was evaluated by administering it intraperitoneally(50,100,200,and 400 mg/kg),in various vehicles (water,PEG-300 and DMSO),and time intervals(30 min prior,simultaneous and 2 h post).The time dependent protection of gossypin(200 mg/kg in PEG-300;i.p.)was also evaluated using selected biochemical variables(GSH,GSSG,MDA,total antioxidant status,Hb,WBC count,RBC count,glutathione peroxidase,glutathione reductase,and superoxide dismutase)and liver histology.The protection of gossypin by oral route was also evaluated against percutaneously administered SM.Results The protection against systemic toxicity of SM(LD50 8.1 mg/kg)was beRer when gossypin was given with PEG-300 (8.0 folds)than DMSO(5.7 folds).No protection was observed when gossypin was administered with water.Good protection (8.0 folds)was observed when gossypin was administered(200 mg/kg in PEG-300;i.p.)at 30 min prior or simultaneous to SM exposure,but no protection was observed when gossypin was administered 2 h post to SM exposure.A significant weight loss was observed 7 days after SM administration(2 LD50),with a significant increase in RBC and Hb.A significant decrease in total antioxidant status of plasma,liver GSH and GSSG levels,and in the activities of glutathione peroxidase,glutathione reductase and superoxide dismutase was also observed 7 days after SM administration.SM treated mouse liver also showed necrosis.A significant protection was observed when gossypin(200 mg/kg in PEG-300;i.p.)was administered either as a pretreatment(30 min before)or simultaneous treatment,and not as a post treatment(2 h).The protective efficacy of gossypin was better through oral route when administered with DMSO(4.8 folds)than with PEG-300(2.4 folds).No protection was observed when gossypin was administered orally with water.Conclusion Percutaneous administration of SM induces oxidative stress and gossypin can protect it as a prophylactic agent by intraperitoneal or oral routes.
Association of the Apolipoprotein B Gene Polymorphisms With Essential Hypertension in Northern Chinese Han Population
2007, 20(3): 260-264.
Objective To study the association of the apolipoprotein B gene polymorphisms with essential hypertension in Northern Chinese Han population.Methods XbaI and EcoRI polymorphisms of the apolipoprotein B(APOB)gene were genotyped by polymerase chain reaction(PCR)and restriction fragment-length polymorphism(RFLP)method in 503 unrelated hypertensive patients and 490 healthy controls recruited from international collaborative study of cardiovascular disease in Asia (InterAsia).Results The difference in the genotypic distributions could be neglected across the groups.The prevalence of X+allele in healthy controls(4.8%)was less frequent in Chinese,and there was no significant difference in the frequency of the X+allele between cases(5.7%)and controls(P＝0.38).The observed E-allele frequencies were closely similar among groups (5.9%in cases vs 5.0%in controls,P=0.39).Logitstic regression analyses revealed that the lack of association still persisted after adjustment of other environmental factors.Haplotype analysis showed that X-E+ was most frequent and no haplotype could significantly contribute to essential hypertension.Conclusion The APOB gene XbaI and EcoRI polymorphisms are not associated with essential hypertension in the Northern Chinese Han population.Future studies on single nucleotide polymorphisms in larger samples are needed to further investigate the possible contribution of the APOB gene to essential hypertension.