To quantify five specific dietary phytosterols and phytostanols in vegetables and fruits commonly consumed in China.Methods A total of 34 different kinds of vegetables and 33 kinds of fruits were chosen according to the consuming habit of Chinese people.All the samples were purchased from two shops in Beijing.The contents of phytosterols (β-sitosterol,campesterol,stigmasterol,β-sitostanol,and campestanol) were analyzed by GLC method which was established by our laboratory,and the total phytosterols were calculated.Results The total phytosterol content in vegetables ranged 1.1-53.7 mg/100 g edible portion.The highest concentration was found in pea,cauliflower,broccoli,and romaine lettuce.The phytosterol contents in fruits ranged 1.6-32.6 mg/100 g,the highest concentration was found in navel orange,tangerine,and mango.Conclusion The phytosterol contents in vegetables and fruits are not as high as those in edible oils,but because of the large amount of consumption,they also play an important role in increasing the people's phytosterols intake,indicating that increased intake of vegetables and fruits with higher phytosterol contents helps increase the phytosterol intake in China.
To identify the genetic defects of the the adiponectin (APM1) gene that contribute to the development of type 2 diabetes (T2DM) and determine the functional single nucleotide polymorphisms (SNPs) in the APM1 gene associated with T2DM in Han nationality.Methods The APM1 gene 5'-UTR was screened by direct sequencing to identify common polymorphisms.Identified SNPs were genotyped in 585 nondiabetic controls,278 subjects with impaired glucose intolerance (IGT) and 212 patients with T2DM.The functions of SNPs in the regulatory region were assessed by reporter gene assay.Possible association between SNPs and plasma APM1 levels or metabolic parameters was statistically assessed.Results Three SNPs were identified in the APM 1 gene 5'-UTR.A case-control study revealed that SNP-11377 G/C had significant differences in allele frequencies between T2DM patients and nondiabetic controls (G 0.314/C 0.686 vs.G 0.265/C 0.735,P=0.03).Haplotype analysis of three SNPs in the APM1 gene showed that no significant association of haplotypes with T2DM.IGT was detected in the present study.Reporter gene assay showed that SNP did not influence the transcription efficiency in the 3T3-L1 cell line.Conclusion SNP-11377 G/C in the proximal promoter region of the APM1 gene contributes to the development of T2DM in Han nationality but may not be a functional SNP in the APM1 gene.
To examine the effect of hydraulic residence time (HRT) on the performance and stability,to treat dilute wastewater at different operational temperatures in a carrier anaerobic baffled reactor (CABR),and hence to gain a deeper insight into microbial responses to hydraulic shocks on the base of the relationships among macroscopic performance,catabolic intermediate,and microcosmic alternation.Methods COD,VFAs,and microbial activity were detected with constant feed strength (300 mg/L) at different HRTs (9-18 h) and temperatures (10℃-28℃) in a CABR.Results The removal efficiencies declined with the decreases of HRTs and temperatures.However,the COD removal load was still higher at short HRT than at long HRT.Devastating reactor performance happened at temperature of 10℃ and at HRT of 9 h.HRTs had effect on the VFAs in the reactor slightly both at high and low temperatures,but the reasons differed from each other.Microbial activity was sensitive to indicate changes of environmental and operational parameters in the reactor.Conclusion The CABR offers to certain extent an application to treat dilute wastewater under a hydraulic-shock at temperatures from 10℃ to 28℃.
Previous work has showed that excess iron accumulation is harmful to reproduction and even promotes death;however,whether the multiple biological toxicity of iron (Fe) exposure could be transferred to progeny remains unknown.The present study used Caenorhabditis elegans to analyze the multiple toxicities of iron exposure and their possible transferable properties.Methods Three concentrations of iron sulfate solution (2.5μmol/L,75μmol/L,and 200 lamol/L) were used.The endpoints of lifespan,body size,generation time,brood size,head thrash and body bend frequencies,and chemotaxis plasticity were selected to investigate Fe toxicity and its effect on progeny in Caenorhabditis elegans.Results The Fe toxicity could cause multiple biological defects in a dose-dependent manner by affecting different endpoints in nematodes.Most of the multiple biological defects and behavior toxicities could be transferred from Fe-exposed Caenorhabditis elegans to their progeny.Compared to the parents,no recovery phenotypes were observed for some of the defects in the progeny,such as body bend frequency and life span.We further summarized the defects caused by Fe exposure into 2 groups according to their transferable properties.Conclusion Our results suggest that Fe exposure could cause multiple biological defects,and most of these severe defects could be transferred from Fe exposed nematodes to their progeny.
To evaluate the effect of white rot fungus Phanerochaete chrysosporium on removal of gaseous chlorobenzene.Methods Fungal mycelium mixed with a liquid medium was placed into airtight bottles.A certain amount of chlorobenzene was injected into the headspace of the bottles under different conditions.At a certain interval,the concentrations in the headspace were analyzed to evaluate the degradation of chlorohenzene by P.chtysosporium.Results The degradation effects of P chrysosporium on chlorobenzene under different conditions were investigated.The difference in the optimum temperature for the growth of the fungi and chlorobenzene degradation was observed.The data indicated that a lower temperature (28℃) would promote the degradation of chlorobenzene than the optimum temperature for the growth of the fungi (37℃).A low nitrogen source concentration (30 mg N/L) had a better effect on degrading chlorohenzene than a high nitrogen source concentration (higher than 100 mg N/L).A high initial concentration (over 1100 mg/m3) of chlorobenzene showed an inhibiting effect on degradation by P.chrysosporium.A maximum removal efficiency of 95% was achieved at the initial concentration of 550 mg/m3.Conclusion P.chrysosporium has a rather good ability to remove gaseous chlorobenzene.A low nitrogen source concentration and a low temperature promote the removal of chlorohenzene by P.chrysosporium.However,a high initial chlorobenzene concentration can inhibit chlorobenzene degradation.
To study environment-friendly determination of azobenzene in trace amounts using β-cyclodextrin (β-CD)-modified Au electrode.Methods β-CD-modified Au electrode was fabricated with a two-step approach,and then a gold electrode modified with β-CD was used to detect azobenzene by employing Osteryoung square wave voltammetry.Results The modified electrode could detect azobenzene,showing a good linearity between the electrochemical current and concentration.Conclusion Although the electrochemical current is related with concentration,the detection limit is around 1.0×10-10 mol/L.This study may provide a new environment-friendly approach for monitoring water quality.
To study the specific amino acid variation in Nef that may be related to disease progression after infection with HIV-1 subtype B,a predominant strain circulating in China,and to determine whether changes in Nef secondary structure may influence different stages of AIDS development based on the concept that the Nef gene of HIV infection dramatically alter the severity of viral infection and virus replication and disease progression,and that long-term non-progressors (LTNP) of HIV infection are commonly associated with either a deletion of the Nef gene or the defective Nef alleles.Methods The study subjects were divided into LTNP1(n=14),LTNP2 (n=16) and slow progressor (SP,n=19) groups for mutational analysis of the Nef sequence.The data were obtained by using Bioedit,MEGA,Anthewin and SAS software.Results Residues in Nef TA48/49 and K151 occurred more frequently in the LTNP group while AA48/49 was more frequendy observed in the SP group.Of the differences observed in the secondary structure comparison using Nef consensus sequences of these three groups,one was roughly corresponding to the Nef48/49 mutation site.Conclusion TA48/49,K151,and A48/49 in the Nef gene might be associated with the different stages of HIV infection,and there may be a link between the Nef secondary structure and the progression of HIV-1 infection.
To determine the in vitro possible clastogenic and cytotoxic activities of Uh,a rigida crude extracts (UP,E),and identify their antigenotoxic and protective effects on chemotherapeutic agent mitomycine-C (MMC).Methods Anti-clastogenic and anti-genotoxic activities of Ulva rigida crude extracts (URE) were studied using chromosome aberration (CA),sister chromatid exchange (SCE),and micronuclei (MN) tests in human lymphocytes cultured in vitro.Results The chromosome aberration,sister chromatid exchange or micronuclei tests showed that URE at concentrations of 10,20,and 40 μg/mL had no clastogenic activity in human lymphocyte cell culture.Three doses of URE significantly decreased the number of chromosomal aberrations and the frequencies of SCE and MN when compared with the culture treated with MMC (P＜0.0001).Conclusion Although URE itself is not a clastogenic or cytotoxic substance,it possesses strong antigenotoxic,anti-clastogenic,and protective effects on MMC in vitro.
To detect the response of lymphocytes to radiation in untreated breast cancer patients with three different genetic assays.Methods Blood samples were collected from 25 untreated patients and 25 controls.Each blood sample was divided into two parts:one was irradiated by 3-Gy X-ray (irradiated sample),the other was not irradiated (non-irradiated sample).The radiosensitivity of lymphocytes was assessed by comet assay,cytokinesis-block micronucleus (CBMN) assay and 6-TG-resistant cells scored (TG) assay.Results The baseline values of micronucleated cell frequency (MCF) and micronucleus frequency (MNF) in the patients were significantly higher than those in the controls (P＜0.01),and 3-Gy X-ray induced genetic damage to lymphocytes in the patients increased significantly as compared with that in the controls as detected with the three genetic assays (P＜0.01).The proportion of radiosensitive cases in the patient group was 48% for the mean tail length (MTL),40% for the mean tail moment (MTM),40% for MCE 44% for MNE and 48% for mutation frequencies of the hprt gene (Mfs-hprt),respectively,whereas the proportion of radiosensitive cases in the control group was only 8% for all the parameters.Conclusion The difference in the lymphocyte radiosensitivity between the breast cancer patients and the controls is significant.Moreover,there are wide individual variations in lymphocyte radiosensitivity of patients with breast cancer.In some cases,the radiosensitivity of the same patient may be different as detected with the different assays.It is suggested that multiple assays should be used to assess the radiosensitivity of patients with breast cancer before therapy.
To study the activities of interleukin (IL)-2 and granulocyte-macrophage colony-stimulating factor (GM-CSF) (hIL-2/mGM-CSF).Methods SOE PCR was used to change the linker of the fusion protein for higher activities.The fusion protein was expressed in Escherichia coli (E.coli) BL21 (DE3) in inclusion body (IB) form.After IB was extracted and clarified,it was denatured and purified by affinity chromatography.The protein was refolded by dilution in a L-arginine refolding buffer and refined by anion chromatography.The protein activity was detected by cytokine-dependent cell proliferation assay Results The expression of hlL-2/mGM-CSF in E.coil yielded approximately 20 mg protein/L culture and the purity was about 90%.The specific activities of IL-2 and GM-CSF were 5.4×106 IU/mg and 7.1×106 IU/mg,respectively.Conclusion This research provides important information about the anti-tumor activity of hIL-2/mGM-CSF in vivo,thus facilitating future clinical research on hIL-2/mGM-CSF used in immune therapy.
To examine the relationship between occurrence of hypedipidemia,plasma homocysteine and polymorphisms of methylenetetra hydrofolate reductase (MTHFR) gene and methionine synthase (MS) gene.Methods A total of 192 hyperlipidemia patients were selected and divided into hypercholesterolemia group,hypertriglyceridemia group,and combined hyperlipidemia group.Another 208 normal individuals were selected as control.Total plasma homocysteine (tHcy)concentration was measured by high-performance liquid chromatography (HPLC).Lipid profiles were measured for all subjects.The polymorphisms of MTHFR gene C677T and MS gene A2756G were analyzed by PCR-RFLP.Results The tHcy concentration in the combined hyperlipidemia patients was significantly higher than that in the control (15.95 μmol/L vs 13.43 μmol/L,P＜0.05).The prevalence of hyperhomocysteinemia (Hhcy) in the combined hyperlipidemia group was significantly higher than that in the control (42.2% vs 23.0%,P=0.015),with the odds ratio (OR) of 3.339 (95%CI:1.260-8.849).The hyperlipidemia patients with Hhcy had a higher concentration of total cholesterol (TC) than that in the normal they patients (5.67±0.95 mmol/L vs 5.47±0.92 mmol/L,P=0.034).There was no significant difference in genotype or allele frequencies of MTHFR C677T between the hyperlipidemic and control groups.The hyperlipidemia patients with MTHFR CT/TT genotype had a higher concentration of triglyceride (TG) than those with CC genotype (2.24±1.75 mmol/L vs 1.87±0.95 mmol/L,P＜0.05).Individuals with CT/TT genotype had a higher concentration of tHey than those with 677CC genotype both in the hyperlipidemia group (12.61±1.24 μmol/L vs 11.20±1.37 μmol/L,P＜0.05) and in the control group (14.04±1.48 μmol/L vs 12.61±1.24 μmol/L,P＜0.05).The percentage of MS 2756 GG/AG genotype in the combined hyperlipidemia group was significantly higher than that in the control (26.7% vs 13.0%,P=0.012),with the OR of 3.121 (95%CI:1.288-7.651).The hyperlipidemia patients with MS 2756AG/GG genotype had a higher concentration of TC (5.87±0.89 mmol/L vs 5.46±0.93 mmol/L,P＜0.05) and LDL-C (3.29±0.81 mmol/L vs 2.94±0.85 mmol/L,P＜0.05)than those with AA genotype.However,individuals with 2756AG/GG genotype showed no significant difference in tHcy among those with AA genotype.Conclusion Hhcy and MS A2756G mutation may be the risk factors for combined hyperlipidemia.Further study is needed to confirm the role of Hhcy and MS A2756G mutation in the development of hyperlipidemia.
To review the clinical features and laboratory investigations of ciguatera patients in Hong Kong between 2004 and 2007 in order to show the timely sampling of implicated fish from ciguatera victims and application of validated mouse bioassay for confirming suspected clinical cases of ciguatera.Methods Diagnosis of the ciguatera victims was based on history of coral fish consumption and clinical presentations stated in official guidelines for clinical diagnosis of ciguatera fish poisoning in Hong Kong.Food remnants of coral fish samples were collected swiftly from ciguatera victims between 2004 and 2007 for ciguatoxins (CTXs) analysis.Results Major clinical symptoms in ciguatera patients included gastrointestinal and neurological effects including limb numbness and diarrhoea,which developed at 0.5 to 15 hours after consumption of fish.In most cases,neurological symptoms were more common than gastrointestinal symptoms.A broad range of attack rate (10%-100%) was observed in each ciguatera outbreak.Validated mouse bioassay on ether extracts of the food remnant samples confirmed that all were CTXs-positive (＜0.5-4.3 MU/20 mg ether extract) and directly linked to the corresponding ciguatera cases.Conclusion Consistency between clinical and laboratory analysis for ciguatera poisoning illustrates the application of laboratory mouse bioassay in a timely fashion for confirming ciguatera poisoning cases and implementing effective public health measures.With further improvement in laboratory techniques,features of ciguatera fish poisoning cases can be better defined.Further studies are needed to determine the risk of each class of CTXs (Pacific-,Indian-and Caribbean-CTXs) in Hong Kong.
To study the association between the rs7566605 variant of INSIG2 and obesity-related phenotypes in Chinese children and adolescents.Methods The study sample consisted of two independent cohorts of Chinese children and adolescents.Anthropometric indices,lipids,blood pressure,fasting glucose,insulin and percentage of fat mass were determined.PCR with restriction fragment length polymorphism analysis was performed for genotyping the rs7566605 variant.Results In each of the two independent cohorts,no significant association was observed between rs7566605 and obesity under additive,dominant or recessive model.We also did not detect any difference in the genotype frequency between all the obese children and controls.Furthermore,we did not find evidence of an association between body composition indices and metabolic phenotypes in all children.However,the triglyceride level of CC homozygotes was significantly higher than that of GG+GC genotypes in obese children (P=0.022).Additionally,we observed a non-significant trend of severe obesity in a post-hoc test.Conclusion INSIG2 rs7566605 variant is not associated Chinese childhood obesity in two independent cohorts.Further study is needed to verify the effect of rs7566605 on triglyceride in obese children.