2006 Vol. 19, No. 4
To evaluate the molluscicidal activities of three Chinese plants N. Indicum Mill, P. Stenoptera DC, and R.japonicum Houtt, and to clarify the molluscicidal mechanism. Methods N-butanol extracts and water extracts of the three plants were obtained. The reactions of EST isozyme, glycogen and total protein of snails to the plant extracts were studied.Results EST electrophoresis showed that EST was an important antidotal enzyme system and reacted strongly to environment.EST changed greatly during the whole exposure period so that it could be viewed as a pathological index of toxicity. Extracts decreased the glycogen content of the snails'soft tissues greatly, and also the protein content. Conclusion All extracts show strong molluscicidal activity. The LD50 value of the water extract of N. Indicum Mill is as low as 13.2 mg/L. EST can be viewed as a pathological index of toxicity. The energy metabolism abnormity is the key reason for the molluscicidal activities. The biochemical mechanism needs further research.
In order to investigate the pharmacological properties of Ginkgo biloba extract (GBE) on improving blood circulation, the regulating action of GBE and quercetin (a main flavonoid ingredient in GBE) on thrombomodulin (TM)expression and tissue-type plasminogen activator (t-PA) secretion was studied. Methods Using flow cytometer and gel image system respectively, we evaluated the TM expression and the t-PA secretion by human umbilical vein endothelial cells (HUVECs) in vitro. Results The increase of TM expression on HUVECs surface was induced by GBE rather than quercetin in a dose- and time-dependent manner. Both GBE and quercetin increased the t-PA release significantly.Conclusion The effect of GBE on improving blood circulation may be partly attributed to its promoting TM expression and t-PA secretion by endothelial cells, and quercetin participated in the effect of GBE on t-PA secretion. However, the action of GBE on increasing TM expression needs further study.
2006, 19(4): 254-261.
To determine the metal contents of date palm (Pheonix dactylifera) samples in dry season from Denizli-Turkey for investigation of heavy metal-polluted traffic. Method The levels of iron, copper, zinc, lead, cadmium,nickel, chromium, and manganese ions in the leaves of thirty five date palm (Pheonix dactylifera) samples collected from various levels of traffic in the streets of Denizli-Turkey were determined by graphite furnace or flame atomic absorption spectrometry. The wet, dry, and microwave digestion procedures for the date palm (Pheonix dactylifera) leaves were compared.The accuracy of the digestion procedures was checked using a standard reference material (IAEA-336 Lichen, SRM). Results Microwave digestion procedure for the leaves was preferred because it was more proper with respect to both time and recovery than dry and wet digestion. The levels of the heavy metal ions investigated were the highest on the samples from high traffic level. Also correlations between metal levels and traffic volume for all the metals were investigated. Conclusion In the light of our findings, the date palm (Phoenix dactylifera) leaves are suitable as a biomonitor for atmospheric heavy metal-polluted traffic. Significant correlations can be obtained between traffic levels and heavy metal concentrations.
To study the characterization apportionment of main ion concentrations of PM10 under the influence of trajectory wind direction in London. Methods PM10 samples from 1 May 1995 to 30 October 1995 of Oxford Street of Central London were collected, the metals and anions of which were measured using atomic absorption spectrometry (AAS) and ion chromatography (IC). Composite trajectories representative of the air mass arriving in London at the same period were calculated based on basic routine back trajectories from the British Atmospheric Data Centre (BADC). Results Concentration apportionments of main ions were similar when the trajectory was plotted back at 6 h, 12 h, and 24 h, some were obviously different. Mg, Ba, Pb, and Cu had similar peak apportionments at the area 180°-320°, but Zn and Ni at the area of 90°-270°, NO3- and SO42- at the area of 100°-220°. Cl- concentration peak apportionment was at the area of 220°-300°, which showed that Cl- mainly came from the North Sea. Conclusion Trajectory wind direction has important effect on ion concentration apportionment of PM10 in London. The ions have similar concentration peak apportionments or their correlation coefficients are statistically significant.
To investigate the effect of ambient air pollution on human health and the subsequent disability-adjusted life years (DALYs) lost in Shanghai. Methods We used epidemiology-based exposure-response functions to calculate the attributable number of cases due to air pollution in Shanghai in 2000, and then we estimated the corresponding DALYs lost in Shanghai based on unit DALYs values of the health consequences. Results Ambient air pollution caused 103 064 DALYs lost in Shanghai in 2000. Among all the health endpoints, premature deaths and chronic bronchitis predominated in the value of total DALYs lost. Conclusion The air pollution levels have an adverse effect on the general population health and strengthen the rationale for limiting the levels of air pollution in outdoor air in Shanghai.
To study the effect of terephthalic acid (TPA) on lipid metabolism in Sprague-Dawley (SD) rats. Methods Five groups of SD rats that ingested 0%, 0.04%, 0.2%, 1%, and 5% TPA, respectively, were included in a 90-day subchronic feeding study.Effects of TPA on levels of serum protein, total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL), total antioxidative capability (T-AOC), superoxide dismutase (SOD) and malondialdehyde (MDA) were observed. Urine samples were collected and analyzed for concentration of ion. Results TPA decreased the level of serum T-AOC in a dose dependent manner. The contents of serum and bladder MDA significantly decreased in 1% and 5% TPA ingestion groups. Serum CuZn superoxide dismutase (CuZnSOD) lowered in groups of 0.2%, 1%, and 5% TPA. TPA subchronic feeding had no significant influences on serum TC, LDL or HDL, but increased serum TG, TP and ALB after administration of 0.04% and/or 0.2% TPA. Concentrations of urinary Ca2+, Mg2+, Na+,and K+ were elevated in 1% and 5% TPA groups. Conclusion Antioxidative potential decreased after TPA exposure. MDA increase in serum and bladder tissues was one of the most important reactions in rats which could protect themselves against TPA impairment.The decrease of serum CuZnSOD was related to the excretion of Zn2+.
To study the alterations of FHIT gene and P16 gene in malignant transformed human bronchial epithelial cells induced by crystalline nickel sulfide using an immoral human bronchial epithelial cell line, and to explore the molecular mechanism of nickel carcinogenesis. Methods 16HBE cells were treated 6 times with different concentrations of NiS in vitro, and the degree of malignant transformation was determined by assaying the anchorage-independent growth and tumorigenicity. Malignant transformed cells and tumorigenic cells were examined for alterations of FHIT gene and P16 gene using RT-PCR, DNA sequencing, silver staining PCR-SSCP and Western blotting. Results NiS-treated cells exhibited overlapping growth. Compared with that of negative control cells, soft agar colony formation efficiency of NiS-treated cells showed significant increases (P＜0.01) and dose-dependent effects. NiS-treated cells could form tumors in nude mice, and a squamous cell carcinoma was confirmed by histopathological examination. No mutation of exon 2 and exons 2-3, no abnormal expression in p16 gene and mutation of FHIT exons 5-8 and exons 1-4 or exons 5-9 were observed in transformed cells and tumorigenic cells. However, aberrant transcripts or loss of expression of the FHIT gene and Fhit protein was observed in transformed cells and tumorigenic cells. One of the aberrant transcripts in the FHIT gene was confirmed to have a deletion of exon 6, exon 7, exon 8, and an insertion of a 36 bp sequence replacing exon 6-8. Conclusions The FHIT gene rather than the P16 gene, plays a definite role in nickel carcinogenesis. Alterations of the FHIT gene induced by crystalline NiS may be a molecular event associated with carcinogen, chromosome fragile site instability and cell malignant transformation. FHIT may be an important target gene activated by nickel and other exotic carcinogens.
To investigate the apoptosis-inducing effect of endogenous nitric oxide (NO) suppression in gastric cancer cells and its mechanisms. Methods Apoptosis of gastric cancer cells was detected by flow cytometry. Expression of phosphorylated FKHRL1 (thr-32, ser-253) and FKHRL1 in gastric cancer cells was analyzed using Western blotting.Immunofluorescence assay was performed to localize the intracellular phosphorylated FKHRL1 (thr-32, ser-253) and FKHRL1.Transfection of FKHRL1-HA wild type and mutant FKHRL1-HA T32A constructs was performed by lipofectamine plus reagent. NO generation was determined by Griess reaction. Results Gastric cancer cells were significantly apoptotic after treatment with NG-monomethyl-L-arginine (L-NMMA, a nitric oxide synthase inhibitor), compared with the control (P＜0.01).The apoptosis of gastric cancer cells induced by L-NMMA was dose-dependent and time-independent. However, the Z-DEVD-fmk, a caspase-3, 6, 7, 8, 10 inhibitor, did not prevent the apoptosis. The immunofluorescence assays showed that FKHRL1 protein was strongly expressed in the nucleu and p-FKHRL1 thr-32 protein was strongly expressed in the cytoplasm of SGC-7901 cells when endogenous nitric oxide generation was blocked by L-NMMA, but no change in FKHRL1 ser-253phosphorylation. Nevertheless, ROCK protein was strongly expressed in p-FKHRL1 thr-32-positive SGC-7901 cells. The wortmannin, an inhibitor of phosphoinositol-3-OH kinase (PI3K), did not block the phosphorylated FKHRL1 thr-32 protein induced by L-NMMA. However, Y-27632, a specific inhibitor of the protein kinase ROCK, significantly blocked apoptosis induced by phosphorylated FKHRL1 thr-32 (P＜0.01), which was mediated by L-NMMA. A significant decrease in NO generation (P＜0.01) and a significant increase in apoptosis (P＜0.01) were observed when FKHRL1-HA wild-type cells were transfected, which caused increased FKHRL1 thr-32 phosphorylation. Conclusions L-NMMA triggers gastric carcinoma cell apoptosis, possibly by promoting FKHRL1 thr-32 phosphorylation and initiating signal of FKHRL1 to ROCK kinase. This apoptotic signaling process is PI3K/Akt as well as caspase-3 independent.
To investigate the relationship between trans, trans-muconic acid (ttMA) as benzene metabolite of occupational workers and benzene concentration in air. Methods A rapid and sensitive high-performance liquid chromatography was developed to determine the level of urinary ttMA. ttMA was extrated from urinary samples in liquid-liquid phase a ODS (2) (5u) column (Φ4.6 mm× 150 mm) and detected at wavelength 264 nm in a UV detector using vanillic acid as an internal standard. The mobile phase was acetaticacid/tetrahydrofuran/methanol/water (v/v, 1:2:10:87). The method was validated with 56 urine samples collected from occupationally benzene-exposed individuals. Results A correlation coefficient (r = 0.9963 ) was found for ttMA ranging 0.10-10.00 μg/mL. The limit of detection was 0.10 μg/mL. The recovery and reproducibility were generally over 90%. There was a positive correlation between ttMA and benzene level in air. The equation was Y=0.859+0.108C (before work, r=-0.6200) or Y=1.980+0.179C (after work, r=0.7930). Conclusion This method can be used to determine and control the level of urinary ttMA in those who are occupationally exposed to benzene.
Bioremediation technology has gained importance because microbes could be the convenient source of bio-absorption/bioaccumulation of metals from effluent streams. Methods The nickel-resistant bacterial isolates (NiRBI)were selected from various bacterial isolates from industrial effluent and grown in nutrient broth containing different concentrations of nickel sulfate (0.3-3.0 mmol/L) and their capability of accumulating metal from the medium. Results Well-defined growth of NiRBI was observed in the medium containing up to 2.5 mmol/L of nickel. The isolate was identified using 16S rRNA and closely related to Pseudomonas fragi. Maximum accumulation of nickel (0.59 mg/g dry weight of bacterial cells) was observed when NiRBI was grown in media containing 2 mmol/L of nickel. The protein profile of the NiRBI cellular extract by SDS-PAGE showed two metal stress-induced proteins of molecular weight 48 KD and 18 KD with a simultaneous down regulation of four proteins of 46.7 KD, 42.2 KD, 19.7 KD, and 4.0 KD. Conclusion 48 KD and 18 KD proteins play a role in metal resistance mechanism by NiRBI.
To investigate the effect of selenium supplementation on the selenium status and selenoenzyme, especially the activity and mRNA expression of type 1 deiodinase (D1) in mice with excessive iodine (EI) intake and to explore the mechanism of selenium intervention on iodine-induced abnormities. Methods Weanling female BALB/c mice were given tap water or 3 mg/L of iodine or supplemented with 0.5 mg/L or 1.0 mg/L of selenium in the presence of excessive iodine for 5months. Selenium status, thyroid hormone level, hepatic and renal D 1 activity and mRNA expression were examined. Results Excessive iodine intake significantly decreased the selenium concentration in urine and liver, and the activity of glutathione peroxidase (GSH-Px) in liver. Meanwhile, serum total T4 (TT4) increased while serum total T3 (TT3) decreased. Hepatic D1enzyme activity and mRNA expression were reduced by 33% and 86%, respectively. Renal D1 enzyme activity and mRNA were reduced by 30% and 55%, respectively. Selenium supplementation obviously increased selenium concentration, activity of GSH-Px and D1 as well as mRNA expression of D1. However, increasing the supplementation of Se from 0.5 to 1.0 mg/L did not further increase selenoenzyme activity and expression. Conclusion Relative selenium deficiency caused by excessive iodine plays an essential role in the mechanism of iodine-induced abnormalities. An appropriate dose of selenium supplementation exercises a beneficial intervention.
To investigate how acetamiprid, a new insecticide, affects the activity of superoxide dismutase (SOD),catalase (CAT), and ATPase and the SOD isozyme patterns in two G- bacteria, E. coli K12 and Pse. FH2, and one G+ bacterum,B. subtilis. Methods The SOD, CAT, and ATPase specific activities of cell lysates were determined spectrophotometrically at 550 nm, 240 nm, and 660 nm, respectively, with kits A001, A016, and A007. SOD isozyme patterns were detected by native PAGE analysis. Results SOD and CAT activities in the tested bacteria increased significantly in a concentration-dependent manner after different concentrations of acetamiprid were applied. The activity of SOD in B. subtilis and Pse. FH2 was stimulated and reached the highest level after treatment with 100 mg/L acetamiprid for 0.5 h. For Pse. FH2, there was another stimulation of SOD activity after acetamiprid application for about 8.0 h and the second stimulation was stronger than the first.The stimulation by acetamiprid showed a relative lag for E. coli K12. Acetamiprid seemed to exhibit a similar effect on CAT activity of the two G- bacteria and had an evident influence on ATPase activity in the three bacteria within a relatively short period. Only one SOD isozyme was detectable in Pse. FH2 and B. subtilis, while different isozyme compositions in E. coli could be detected by native PAGE analysis. Conclusion Acetamiprid causes a certain oxidative stress on the three bacteria which may not only elevate SOD and CAT activities but also generate new SOD isozymes to antagonize oxidative stress.However, this oxidative stress lasts for a relatively short time and does not cause a long-term damage.
To study the regulating effects of a novel CpG oligodeoxynuleotide and the synergistic effect of chitosan-nanoparticles (CNP) with CpG on immune responses of mice, which were used to develop a novel immunoadjuvant to boost immune response to conventional vaccines. Methods A novel CpG ODN containing 11 CpG motifs was synthesized and its bioactivities to stimulate the proliferation of lymphocytes of pig in vitro were detected. Then it was entrapped with CNP prepared in our laboratory by the method of ionic cross linkage, and immunized Kunming mice were co-inoculated with paratyphoid vaccine. The peripheral blood was collected weekly from the tail vein of inoculated mice to detect the contents of IgG, IgA, IgM, and specific antibody against salmonella as well as the levels of interleukin-2 (IL2), IL-4, and IL-6 by SABC-ELISA assay. The numbers of leucocytes, monocytes, granuloytes, and lymphocytes were calculated separately using the routine method. The experimental mice were orally challenged with virulent salmonella 35 days after inoculation. Results This CpG ODN could remarkably provoke the proliferation of lymphocytes of pig in vitro in contrast with the control (P＜0.05).Compared with those of the control, immunoglobulins, including IgG,IgA, IgM, and specific antibodies to paratyphoid vaccine,increased significantly in sera from the CpG or CpG-CNP-vaccinated mice (P＜0.05). IL-2, IL-4, and IL-6 increased remarkably in sera from immunized mice (P＜0.05). The leucocytes, monocytes, granuloytes, and lymphocytes of the mice immunized with CpG or CpG-CNP were also increased in number (P＜0.05). After the challenge, these immunity values were elevated in the mice vaccinated with CpG or CpG-CNP. The immunized mice all survived, while the control mice fell ill with evident lesions with diffuse hemorrhage in stomach, small intestine, and peritoneum. Conclusions CpG ODN entrapped with CNP is a promising effective immunoadjuvant for vaccination, which promotes humoral and cellular immune responses, enhances immunity and resistance against salmonella by co-administration with paratyphoid vaccine.
To investigate the association of angiotensin converting enzyme (ACE) gene insertion/deletion (I/D)polymorphism with type 2 diabetes mellitus (T2DM). Methods Two hundred and nine patients with T2DM diagnosed based on the criteria for diabetes mellitus in 1999 by WHO and 221 controls were recruited from general population of Dongcheng District in Beijing. All subjects were genotyped for the I/D polymorphism of ACE gene by PCR-fragment length polymorphism (FLP) assay. Blood pressure, levels of plasma glucose, lipids and serum insulin were determined. Body mass index (BMI),waist-hip ratio (WHR) and homeostasis model assessment-insulin resistance index (HOMA-IR) were calculated. Results The genotype frequencies for ACE genes DD, ID, and Ⅱ were 19.1%, 42.1%, and 38.8% in patients, respectively, and 9.6%,49.4%, and 41.0% in controls, respectively. The ACE DD genotype frequency was significantly higher in patients than in controls (χ2=7.61, P=0.022). Multivariate logistic regression analysis showed that the ACE DD genotype was a risk factor for T2DM, with the OR of 2.35 (95% CI 1.17-4.71) adjusted for age, sex, BMI, WHR, blood pressure, and serum cholesterol levels.Conclusion The ACE DD genotype is associated with the increased susceptibility to type 2 diabetes mellitus.
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