Objective This work is an evaluation of the efficiency of a sand-gravel or unwoven fabric bed system and Lolium perenne Lam as plant biofiltter in the reduction of solids and nutrients removal from aquaculture discharge water. Methods The first step consisted of the collection of wastewater in the tank and the distribution at three different hydraulic loading regimes (0.5, 1, 1.5L/hour) to the different experimental systems. The second step was to evaluate the performance of the different systems. The first system consisted of a bucket filled with a substrate of sand/gravel (20 cm in depth), on the bottom of which was a 80 mesh/inch2 of nylon (S1); the second was similar, but was planted with Lolium perenne lam (S2); the third was planted with a grass plate consisting of 7 layers of unwoven fabric planted with L. perenne (S3). Results The second system showed the best performance in reducing solids as well as in nutrients (TN, TP, and COD) reduction. The removal rates for TS, TN, and TP were negatively correlated with the loading regimes, with 0.5 L/hour being the most efficient and thus taken as the reference. Conclusions Solids management using a sand/gravel substrate as bed culture and Lolium perenne L. as plant biofilter has proved to be an efficient technique for solids reduction with low operating cost. This grass plays an important role in wastewater eco-treatment by absorbing dissolved pollutants (TAN) as nutrients for its growth.
Objective To analyze the impact of depletion of the twin arginine translocation (TAT) system on virulence and physiology of Yersinia enterocolitica for a better understanding of its pathogenicity. Methods We constructed a △tatC::SpRmutant of Yersinia enterocolitica by P1 phage mediated transduction using Escherichia coli K-12 △tatC::SpR strain as a donor. Results A P1-mediated genetic material transfer was found between the two species of enterobacteria, indicating a great potential of acquisition of antibiotic resistance in emergency of a new threatening pathogen by genetic material exchanges.Periplasmic trimethylamine N-oxidase reductase activity was detected in the wild type Y. enterocolitica strain and translocation of this enzyme was completely abolished by the △tatC::SpR mutation. In addition, the △tatC::SpR mutation showed a pleiotropic effect on the metabolism of Y. enterocolitica. However, the tat mutation did not seem to affect the mobility and virulence of Y.enterocolitica under the conditions used. Conclusion Unlike other pathogenic bacteria studied, the TAT system of Y.enterocolitica might play an important role in the pathogenic process, which is distinct from other pathogens, such as Pseudomonas aeruginosa and enterohemorrhagic E. coli O157:H7.
Objective To characterize the heterotrophic nitrifying bacteria. Methods The bacteria were isolated from membrane bioreactor for treating synthetic wastewater using the method newly introduced in this study. Fluorescence in situ hybridization (FISH) was used to validate the nonexistence of autotrophic ammonia oxidizers and nitrite oxidizers. Batch tests were carried out to investigate the capability of heterotrophic nitrification by the pure culture. Phylogenetic analysis of the pure culture was performed. Results A heterotrophic nitrifier, named Bacillus sp. LY, was newly isolated from the membrane bioreactor system in which the efficiency of TN removal was up to 80%. After 24-day, incubation, the removal efficiency of COD by Bacillus sp. LYwas 71.7%. The ammonium nitrogen removal rate after assimilation nearly ceased by Bacillus sp. LYwas 74.7%.The phylogenetic tree of Bacillus sp. LY and the neighbouring nitrifiers were given. Conclusions The batch test results indicate that Bacillus sp. LY can utilize the organic carbon as the source of assimilation when it grows on glucose and ammonium chloride medium accompanying the formation of oxidized-nitrogen. It also can denitrify nitrate while nitrifying. Bacillus sp. LY may become a new bacterial resource for heterotrophic nitrification and play a bioremediation role in nutrient removal.
Objective To investigate the effect of X-rays on expression of caspase-3 and p53 protein in EL-4 cells and its implications in induction of apoptosis and polyploid cells. Methods Mouse lymphoma cell line (EL-4 cells) was used.Fluorescent staining and flow cytometry analysis were employed for measurement of protein expression, apoptosis, cell cycle,and polyploid cells. Results The expression of caspase-3 protein increased significantly at 8 h and 12 h, compared with that of sham-irradiated control (P＜0.05, respectively) and the expression of p53 protein increased significantly at 2, 4, 8, 12, and 24 h, compared with that of sham-irradiated control (P＜0.05-P＜0.01) in EL-4 cells after 4.0 Gy X-irradiation. Apoptosis of EL-4 cells was increased significantly at 2, 4, 8, 12, 24, 48, and 72 h after 4.0Gy exposure, compared with that of sham-irradiated control (P＜0.05-P＜0.001). G2 phase cells were increased significantly at 4, 8, 12, 24, 48, and 72 h (P＜0.05-P＜0.001). However, no marked change in the number of 8 C polyploid cells was found from 2 to 48 h after 4.0 Gy exposure. Conclusion The expressions of caspase-3 and p53 protein in EL-4 cells are induced by X-rays, which might play an important role in the induction of apoptosis, and the molecular pathway for polyploid formation might be p53-independent.
Objective To estimate the biological exposure limit (BEL) using benchmark dose (BMD) based on two sets of data from occupational epidemiology. Methods Cadmium-exposed workers were selected from a cadmium smelting factory and a zinc product factory. Doctors, nurses or shop assistants living in the same area served as a control group. Urinary cadmium (UCd) was used as an exposure biomarker and urinary β2-microgloburin (B2M), N-acetyl-β-D-glucosaminidase (NAG) and albumin (ALB) as effect biomarkers. All urine parameters were adjusted by urinary creatinine. Software of BMDS (Version 1.3.2, EPA.U.S.A) was used to calculate BMD. Results The cut-off point (abnormal values) was determined based on the upper limit of 95% of effect biomarkers in control group. There was a significant dose response relationship between the effect biomarkers (urinary B2M, NAG, and ALB) and exposure biomarker (UCd). BEL value was 5 μg/g creatinine for UB2M as an effect biomarker, consistent with the recommendation of WHO. BEL could be estimated by using the method of BMD. BEL value was 3 μg/g creatinine for UNAG as an effect biomarker. The more sensitive the used biomarker is, the more occupational population will be protected. Conclusion BMD can be used in estimating the biological exposure limit (BEL). UNAG is a sensitive biomarker for estimating BEL after cadmium exposure.
Objective To develop a high-throughput screening assay for Farnesoid X receptor (FXR) agonists based on mammalian one-hybrid system (a chimera receptor gene system) for the purpose of identifying new lead compounds for dyslipidaemia drug from the chemical library. Methods cDNA encoding the human FXR ligand binding domain (LBD) was amplified by RT-PCR from a human liver total mRNA and fused to the DNA binding domain (DBD) of yeast GAL4 of pBIND to construct a GAL4-FXR (LBD) chimera expression plasmid. Five copies of the GAL4 DNA binding site were synthesized and inserted into upstream of the SV40 promoter of pGL3-promoter vector to construct a reporter plasmid pG5-SV40 Luc. The assay was developed by transient co-transfection with pG5-SV40 Luc reporter plasmid and pBIND-FXR-LBD (189-472) chimera expression plasmid. Results After optimization, CDCA, a FXR natural agonist, could induce expression of the luciferase gene in a dose-dependent manner, and had a signal/noise ratio of 10 and Z'factor value of 0.65. Conclusion A stable and sensitive cell-based high-throughput screening model can be used in high-throughput screening for FXR agonists from the synthetic and natural compound library.
Objective To investigate the therapeutic potential of tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL), a member of the TNF superfamily, and to analyze TRAIL-induced apoptosis in Jurkat cells. Methods Expression of TRAIL receptors (DR4 and DR5) was detected by reverse transcriptase-polymerase chain reaction (RT-PCR). Cytotoxic effects were determined by colony formation assay and a cell counting kit. The effects of recombinant TRAIL on apoptosis of Jurkat cells were determined by DNA fragmentation (DNA ladder) and PI staining. Changes in mitochondrial membrane potential were detected with JC-1 fluorescence. Results TRAIL inhibited the proliferation and induced internucleosomal DNA fragmentation (characteristic of apoptosis) and loss of mitochondrial membrane potential. Conclusion Recombinant soluble TRAIL can be used as a therapy for cancer.
Objective To investigate the characteristics of Zn2+ biosorption and the release of cations during the process of Zn2+biosorption by intact cells of Saccharomyces cerevisiae. Methods The batch adsorption test was used to study the biosorption equilibrium and isotherm. Zn2+ concentration was measured with atomic adsorption spectrophotometer (AAS) AAS 6.Vario. Results When the initial concentration of Zn2+ ranged between 0.08 and 0.8 mmol/L, the initial pH was natural (about 5.65), the sorbent concentration was about 1 g/L and the capacity of Zn2+ biosorption was from 74.8 to 654.8 μmol/g. The pH value increased by 0.55-1.28 and the intracellular cations (K+, Mg2+, Na+, Ca2+) of the cells were released during the process of Zn2+ biosorption. Conclusion Ion exchange was one of the mechanisms for Zn2+ biosorption. The biomass of Saccharomyces cerevisiae is a potential biosorbent for the removal of Zn2+ from aqueous solution. More work needs to be done before putting it into practical application.
Objective To study the contamination of total aflatoxins (AFs) in different kinds of foods including corn, peanut, rice,walnut, and pine nut in six provinces and two municipalities in China. Methods A total of 283 samples of corn, peanut, rice,walnut and pine nut were randomly collected from local markets in Fujian, Guangdong, Guangxi, Hubei, Jiangsu, and Zhejiang provinces, as well as in Shanghai and Chongqing municipalities. The samples were ground to which acetonitrile/water solution was added. After filtering, the extract was transferred into a MycoSepTM purifying column and was pressed slowly. Then the purified liquid was derivatized with trifluoroacetic acid (TFA) and assayed using high performance liquid chromatography (HPLC). Results AFs were detected in 70.27% of corn samples, with a mean level of 27.44 μg/kg and the highest level of 1098.36 μg/kg. In peanut, the AFs detection rate was 23.08%, with a mean level of 0.82 μg/kg and the highest level of 28.39 μg/kg.Very few rice samples with AFs were detected. The AFs levels were very low in walnut and pine nut. Conclusion Corn is the food most seriously contaminated with AFs in China. AFB1 is the main aflatoxin which is found as a contaminant in foods.
Objective To isolate human antibodies against hepatitis E virus from phage display library by a new method of panning phage antibody library based on immobilized metal affinity chromatography (IMAC). Methods Phage antibody library was allowed to mix with hex-His tagged expressed HEV specific antigen, NE2, in solution for adequate binding before affinity resin for hex-His was added. The non-specific phage antibodies were removed by extensive washing and the specific bound phage antibodies could then be eluted to infect TG1 or repeat the binding process for subsequent rounds of purification. The specificity of the selected human antibodies were tested by antigen competitive ELISA, human sera blocking ELISA, scFv expression, and sequence analysis. Results His-NE2 specific recombinant phages were successfully enriched after panning procedure. Two individual phage clones, 126 and 138, showed 50% inhibition in NE2 antigen competition ELISA and obvious blocking effect by HEV positive serum in blocking ELISA. Soluble scFv of 126, 138 bound to NE2 specifically. Conclusion Two specific human phage antibodies against hepatitis E virus (HEV) from phage display library were isolated by immobilized metal affinity chromatography. The immobilized metal affinity chromatography applied to phage antibody selection was a helpful supplement to the selection in solution.
Objective To assess the ability of tetrandrine (Tet) to enhance the sensitivity to irradiation and its mechanism in cell lines of human breast cancer p53-mutant MCF-7/ADR, p53-wild-type MCF-7 and human colon carcinoma p53-mutant HT-29 as well as in C26 colorectal carcinoma-bearing BALB/c mice. Methods MCF-7/ADR, HT-29 and MCF-7 cells were exposed to irradiation in the absence or presence of tetrandrine. The effect of Tet on the cytotoxicity of X-irradiation in these three cells was determined and the effect of tetrandrine on cell cycle arrest induced by irradiation in its absence or presence was studied by flow cytometry, Moreover, mitotic index measurement determined mitosis of cells to enter mitosis. Western blotting was employed to detect cyclin B1 and Cdc2 proteins in extracts from irradiated or non-irradiated cells of MCF-7/ADR, HT-29 and MCF-7 treated with tetrandrine at various concentrations. Tumor growth delay assay was conducted to determine the radio-sensitization of tetrandrine in vivo. Results Clonogenic assay showed that tetrandrine markedly enhanced the lethal effect of X-rays on p53-mutant MCF-7/ADR and HT-29 cells and the sensitization enhancement ratio (SER) of tetrandrine was 1.51 and 1.63, but its SER was only 1.1 in p53-wt MCF-7 cells. Irradiated p53-mutant MCF-7/ADR and HT-29 cells were only arrested in G2/M phase while MCF-7 cells were arrested in G1 and G2/M phases. Radiation-induced G2 phase arrests were abrogated by tetrandrine in a concentration-dependent manner in MCF-7/ADR and HT-29 cells,whereas redistribution within MCF-7 cell cycle changed slightly. The proportion of cells in M phase increased from 1.3% to 14.7% in MCF-7/ADR cells, and from 1.5% to 13.2% in HT-29 cells, but 2.4% to 7.1% in MCF-7 cells. Furthermore, the levels of cyclin B 1 and Cdc2 expression decreased after X-irradiation in MCF-7/ADR and HT-29 cells, and the mitotic index was also lower. Tet could reverse the decrease and induce the irradiated cells to enter mitosis (M phase). Endosomatic experiment showed that tetrandrine caused tumor growth delay in irradiated mice. Conclusion Tetrandrine boosts the cell killing activity of irradiation both in vitro and in vivo. Tetrandrine is a potent abrogator for G2 checkpoint control and can sensitize the cells to radiation.
Objective To examine the effect of particulate matter (PM) less than 10 microns in diameter (PM10) and ozone (O3) on daily mortality in Shanghai, China. Methods A generalized additive model with penalized spline function was used to observe the acute effect of PM10 and O3 on daily mortality. Results Higher PMt0 significantly increased the effect of O3 on total mortality,and O3 also increased the effect of PM10 although the estimated increment was statistically insignificant. Conclusion Our findings provide further evidence for the effect of PM10 and O3 on daily mortality.
Objective To establish the association between genetic polymorphisms of HLA-DMA and HLA-DMB and risk of developing trichloroethylene-induced medicamentosa-like dermatitis (TIMLD). Methods Sixty-one cases were medically confirmed to have been affected with TIMLD and 60 controls were selected from exposed workers who were free from TIMLD.The TIMLD cases and controls were similar in terms of age, sex, and duration of exposure. DNA was extracted both from the TIMLD cases and controls, HLA-DMA and HLA-DMB loci were amplified by using Touchdown PCR, and the alleles and genotypes were confirmed by restriction fragment length polymorphism (RFLP) and direct sequencing. Finally, the frequencies of HLA-DMA and HLA-DMB variants were compared between the two groups. Results The results showed that the frequency of HLA-DMA*0101 and HLA-DMB*0103 alleles was significantly increased in TIMLD patients than in controls (71.3% vs. 55.0% for HLA-DMA*0101; P＜0.05) (11.5% vs. 3.3% for HLA-DMB*0103; P＜0.05). In addition, the frequency of HLA-DMA*0102-*0102 homozygous genotype was also significantly higher in the controls than in the patients (25.0% vs.8.2%, P＜0.05), whereas the frequency of heterozygous HLA-DMB *0101-*0102 genotype was lower in the patients in comparison with the controls. Conclusion The polymorphisms of HLA-DM may be associated with the susceptibility to TIMLD.
Objective To evaluate two commercial anti-hepatitis E virus (HEV) IgM kits used for differential diagnosis of acute enteric viral hepatitis. Methods The kit for IgM capture assay, was produced with a recombinant HEV structural protein protecting primates against experimental infection by different HEV genotypes, while the other kit for indirect ELISA was produced with recombinant structural proteins from different HEV genotypes. The serum specimens were taken from 241cases with a confirmed or presumptive diagnosis of hepatitis A and 74 cases with a confirmed or presumptive diagnosis of hepatitis E. Results The sensitivity and specificity of the IgM capture assay kit were 97% and 100%, respectively, and the corresponding values for the other kit were 70% and 78%, respectively. Conclusion The IgM capture assay kit has higher sensitivity and specificity in diagnosing acute enteric viral hepatitis E.