Inhibition of FoxO1 alleviates polycystic ovarian syndrome by reducing inflammation and the immune response.

The aim of this study was to explore the role of forkhead box transcription Factor O1 (FoxO1) in chronic inflammation in polycystic ovary syndrome (PCOS). A PCOS rat model was constructed as an in vivo model by letrozole induction, and granulosa cells (GCs) from PCOS rats were isolated and cultured as an in vitro cellular model. FoxO1 was knocked down by shRNA and siRNA in the PCOS rat model and GCs model, respectively. H&E staining was conducted to evaluate the effect of FoxO1 inhibition on ovarian pathology and dysfunction in PCOS rats. The levels of inflammatory cytokines in the ovaries and uterus of PCOS rats and in GCs were assessed by ELISA. Flow cytometry was used to evaluate the changes in the contents of neutrophils and macrophages in the peripheral blood and spleen of PCOS rats. CCK-8 assays and Annexin V-FITC/PI staining were performed to evaluate the proliferation and apoptosis of GCs. The expression of genes and proteins related to the TLR4/NF-κB/NLRP3 pathway in GCs was determined by RT-qPCR and Western blotting. The results indicated that FoxO1 was highly expressed in PCOS rat model. Inhibition of FoxO1 significantly mitigated the pathological changes and dysfunction in the ovaries of PCOS rats while also suppressing inflammation and fibrosis in the ovaries and uterus. Moreover, knocking down FoxO1 facilitated the restoration of the normal ratio of neutrophils and macrophages in the peripheral blood and spleen of PCOS rats and promoted M2 polarization of macrophages. Additionally, inhibition of FoxO1 promoted the proliferation of GCs and inhibited the inflammatory response in GCs. Furthermore, FoxO1 knockdown inhibited the activation of the NF-κB pathway and the formation of the NLRP3 inflammasome in GCs. In conclusion, inhibition of FoxO1 can alleviate PCOS by inhibiting the TLR4/NF-κB/NLRP3 pathway to reduce inflammation and the immune response.

Response of submerged macrophytes and leaf biofilms to different concentrations of oxytetracycline and sulfadiazine.

Oxytetracycline and sulfadiazine were widely used and they entered the environment through various channels such as domestic sewage, medical wastewater and agricultural wastewater, causing significant ecological risk. To determine the effects of different antibiotic concentrations on submerged macrophytes, Vallisneria natans was exposed to solutions containing different concentrations of oxytetracycline and sulfadiazine (0.1 mg/L、1 mg/L、10 mg/L、50 mg/L). After 20-days exposure, we found that 10 mg/L groups had a significant effect on Vallisneria natans. Under high antibiotic concentrations, the growth of Vallisneria natans was inhibited, chloroplasts were deformed, the chlorophyll content was reduced, and antioxidant enzyme activities, such as superoxide dismutase and glutathione, were increased. There was no significant difference between the control group and groups with low antibiotic concentrations (≤1 mg/L). The N-acyl-l-homoserine lactone concentration tended to increase with increasing antibiotic concentrations. The presence of antibiotics also affected the microbial community structure of biofilms on the submerged macrophytes. For example, the higher the concentration of antibiotics, the higher the proportion of Proteobacteria. These results suggest that high concentrations of oxytetracycline and sulfadiazine can disrupt homeostasis, induce effective Vallisneria natans defense mechanisms and alter biofilms in aquatic ecosystems.

Fertility Outcome and Safety of Ethiodized Poppy Seed Oil for Hysterosalpingography in 1,053 Infertile Patients: A Real-World Study.

Objective: Ethiodized poppy seed oil for hysterosalpingography (HSG) is reported to display some therapeutic effect on infertility, but big a sample-size study under real clinical settings is still lacking to verify the speculation. Thus, this real-world study enrolled 1,053 infertile patients who underwent ethiodized poppy seed oil-based HSG to explore its fertility enhancement value. Method: A total of 1,053 infertile patients who underwent HSG using ethiodized poppy seed oil as the contrast medium were retrospectively analyzed. The live birth rate and 3-, 6-, 12-month and total pregnancy rate were retrieved. Besides, adverse events during and after HSG were recorded. Results: The 3-, 6-, 12-month and total pregnancy rate was 22, 36.8, 50, and 53.8%, respectively. The total live birth rate was 42.7%. Sub-group analyses showed that pregnancy rate was 53.7, 53.8, 54.1, and 62.4% in subgroups of primary infertility patients, secondary infertility patients, infertility patients with fallopian tube disease, and infertility patients with unknown cause, respectively. Meanwhile the live birth rate was 44.3, 41.3, 41.5, and 59.2% in these subgroups, separately. Multivariate logistic regression analysis disclosed that BMI ≥ 24 kg/m2, history of dysmenorrhea, and abnormity of sperm count or motility-related infertility were independently correlated with reduced pregnancy rate and livebirth rate (All Ps < 0.05). Adverse events mainly included pain (20.6%) and interstitial reflux (7.9%), which were mild and tolerable. Conclusion: Ethiodized poppy seed oil for HSG discloses a satisfying fertility outcome with a tolerable safety profile in infertile patients; meanwhile, this effect might be influenced by BMI, history of dysmenorrhea, and paternal abnormity of sperm.

Combined Effects of Allelopathic Polyphenols on Microcystis aeruginosa and Response of Different Chlorophyll Fluorescence Parameters.

Polyphenols are allelochemicals secreted by aquatic plants that effectively control cyanobacteria blooms. In this study, sensitive response parameters (including CFPs) of Microcystis aeruginosa were explored under the stress of different polyphenols individually and their combination. The combined effects on M. aeruginosa were investigated based on the most sensitive parameter and cell densities. For pyrogallic acid (PA) and gallic acid (GA), the sensitivity order of parameters based on the EC50 values (from 0.73 to 3.40 mg L-1 for PA and from 1.05 to 2.68 mg L-1 for GA) and the results of the hierarchical cluster analysis showed that non-photochemical quenching parameters [NPQ, q N, q N(rel) and q CN] > photochemical quenching parameters [YII, q P, q P(rel) and q L] or others [F v/F m, F' v /F' m, q TQ and UQF(rel)] > cell densities. CFPs were not sensitive to ellagic acid (EA) and (+)-catechin (CA). The sensitivity order of parameters for M. aeruginosa with PA-GA mixture was similar to that under PA and GA stress. The quantitative (Toxicity Index, TI) and qualitative (Isobologram representation) methods were employed to evaluate the combined effects of PA, GA, and CA on M. aeruginosa based on cell densities and NPQ. TI values based on the EC50 cells suggested the additive effects of binary and multiple polyphenols, but synergistic and additive effects according to the EC50 NPQ (varied from 0.16 to 1.94). In terms of NPQ of M. aeruginosa, the binary polyphenols exhibited synergistic effects when the proportion of high toxic polyphenols PA or GA was lower than 40%, and the three polyphenols showed a synergistic effect only at the ratio of 1:1:1. Similar results were also found by isobologram representation. The results showed that increasing the ratio of high toxic polyphenols would not enhance the allelopathic effects, and the property, proportion and concentrations of polyphenols played an important role in the combined effects. Compared with cell densities, NPQ was a more suitable parameter as evaluating indicators in the combined effects of polyphenols on M. aeruginosa. These results could provide a method to screen the allelochemicals of polyphenols inhibiting cyanobacteria and improve the inhibitory effects by different polyphenols combined modes.

Quantitative proteomic analysis reveals complex regulatory and metabolic response of Iris lactea Pall. var. chinensis to cadmium toxicity.

Cadmium pollution has become a serious environmental problem. Iris lactea var. chinensis showed strong Cd tolerance and accumulation ability, which has significant potential to be applied for the phytoremediation of Cd-contaminated soil. However, the lack of molecular information on the mechanism of I. lactea response to Cd limited the improvement of phytoremediation efficiency. In this study, label-free proteomics analysis of Cd response in I. lactea showed that there were 163 and 196 differentially expressed proteins (DEPs) in the shoots and roots, respectively. Bioinformatics analysis indicated the DEPs responding to Cd stress mainly involved in signal transduction, ion transport, redox etc., and participate in the pathway of amino acid biosynthesis, lignin biosynthesis, glycerolipid metabolism and glutathione metabolism. Besides, differential expression of seven DEPs was validated via gene expression analysis. Finally, we found that a Cd-induced mannose-specific lectin (IlMSL) from I. lactea enhanced the Cd sensitivity and increased Cd accumulation in yeast. The results of this study will enhance our understanding of the molecular mechanism of Cd tolerance and accumulation in I. lactea and ultimately provide valuable resources for using Cd tolerant genes for developing efficient strategies for phytoremediation of Cd-contaminated soils or limiting Cd accumulation in food crops.

Synthesis of magnetite/lanthanum hydroxide composite and magnetite/aluminum hydroxide composite for removal of phosphate.

Two magnetic adsorbents, magnetite/aluminum hydroxide composite (MAC) and magnetite/lanthanum hydroxide composite (MLC), were successfully synthesized by a simple one-pot method and their phosphate adsorption process was investigated. The properties of synthesized adsorbents were studied using Fourier transform infrared spectroscopy (FTIR), zeta potential, vibrating sample magnetometry (VSM) and X-ray photoelectron spectroscopy (XPS). The adsorption isotherms, adsorption kinetics and the effects of solution pH and dissolved organic carbon (DOC) on the adsorption of phosphate in aqueous solution by MAC, MLC-2, MLC-10 and LMB were investigated to evaluate the difference in phosphate removal efficiency of the magnetic adsorbents and non-magnetic adsorbent. According to the results of this study, MLC-10 had a higher phosphate adsorption capacity (19.34 mg P g-1) than LMB (11.55 mg P g-1), MAC (10.48 mg P g-1) and MLC-2 (8.89 mg P g-1). MLC-10 showed a relative higher partition coefficient (PC) (1.74 mg g-1 µM-1) than other three adsorbents at initial P concentration of 15 mg L-1. Also, MLC-10 was less pH dependent than MAC and had higher phosphate adsorption capacities under different DOC concentrations (0-72 mg L-1) than LMB, MAC and MLC-2. Further, MLC-10 had excellent recyclability due to high magnetism. Electrostatic interaction and the inner-sphere surface complexation were the potential phosphate adsorption mechanisms employed by MLC-10. In summary, MLC-10 is a promising adsorbent for phosphate removal from eutrophication water.

Incidence and Risk Factors of Pressure Injuries in Surgical Spinal Patients: A Retrospective Study.

PURPOSE: The purpose of this study was to measure the incidence and identify risk factors of pressure injury development during the perioperative period in patients undergoing spinal surgery requiring intraoperative positioning in the prone position. DESIGN: Review of medical records. SUBJECTS AND SETTING: The sample comprised 3834 patients; 52.2% (n = 2006) were male and 65.5% (n = 2516) were older than 60 years. Most patients underwent surgery of the lumbosacral spinal segments (43.4%, n = 1667) followed by cervical (32.3%, n = 1241) and thoracic spinal segments (24.2%, n = 932). The study setting was the First Affiliated Hospital of Nanchang University, Jiang XI Province in southeastern China. METHODS: We reviewed charts of patients who underwent spinal surgery requiring intraoperative positioning in the prone position from November 2013 to July 2016. Demographic data, Braden Scale for Pressure Sore Risk cumulative score (measured before preoperative transport), body mass index (BMI), duration of surgery, preoperative time (time between preoperative transport from the inpatient unit to when the operation began), postoperative time (time between when the operation was over and postoperative transport to the inpatient unit), and development of any pressure injury were collected using a standardized form. Factors associated with an increased or decreased likelihood of pressure injury were initially evaluated with χ and independent t tests. Logistic regression was then used to identify potential risk factors for perioperative pressure injury in patients undergoing open spinal surgery requiring placement in the supine position during surgery. RESULTS: One hundred eighty-four of 3840 patients (4.7%) developed pressure injuries. Multivariate analysis indicated that factors associated with intraoperative pressure injury development were older than 60 years (odds ratio [OR] = 1.05, 95% confidence interval [CI] = 1.02-2.17), BMI under 18 kg/m (OR = 2.45, 95% CI = 4.05-5.21), cumulative Braden Scale score 13 or less (OR = 6.59, CI = 2.23-3.98), prolonged preoperative time (OR = 5.99, 95% CI = 3.21-6.12), and prolonged postoperative time (OR = 14.23, 95% CI = 10.23-21.19). CONCLUSIONS: Based on these findings we recommend extending preventive interventions for pressure injury to incorporate the time from preoperative transport to the surgical suite to inpatient care unit following surgery.

Exogenous glutathione increased lead uptake and accumulation in Iris lactea var. chinensis exposed to excess lead.

Long- and short-term hydroponic experiments were conducted to study the effect of different concentrations of exogenous glutathione (GSH) on Pb uptake, translocation, and gene expresses in Iris lactea var. chinensis exposed to excess lead (Pb). Exogenous GSH remarkedly promoted Pb uptake and translocation in long-term (14 d) experiment, and the GSH-dose-dependent increases in shoot and root Pb contents existed obviously when GSH concentrations were lower than 800 mg·L-1. The fresh weight in gradual rise in plants was observed with the increase of exogenous GSH concentration. In short-term (24 h) experiment, Pb contents in roots under Pb with L-buthionine sulfoximine (BSO, a known inhibitor of GSH biosynthesis) treatments were significantly lower than that under Pb exposure alone. The transcript levels of three genes (Ilγ-ECS, IlGS, and IlPCS) involved in GSH synthesis and metabolism, showed no significant change in expression pattern except that upregulation after 24 h of treatment with Pb and GSH in comparison with that of the single Pb treatment. Further, the level of IlGS transcript after exposure for 4 h was much higher than that of Ilγ-ECS and IlPCS transcripts. All these results obtained here suggest that exogenous GSH can increase Pb accumulation, detoxification, and translocation to the shoot.

Iris lactea var. chinensis (Fisch.) cysteine-rich gene llCDT1 enhances cadmium tolerance in yeast cells and Arabidopsis thaliana.

IlCDT1, a cysteine-rich protein, was isolated from Iris lactea var. chinensis (Fisch.) (I. lactea var. chinensis). Its transcription was up-regulated by the exogenous application of Cd. The truncated IlCDT1 (25-54) containing 14 Cys residues confers Cd tolerance to yeast as the intact IlCDT1, indicating that Cys residues are required for Cd tolerance presumably by chelating Cd. When the gene was constitutively expressed in A. thaliana, root length of transgenic lines was longer than that of wild-type under 100 µM or 200 µM Cd stress. However, Cd absorption in wild-type was more than in two trangenic lines under 100 µM Cd exposure. IlCDT1 may directly bind Cd, through chelating Cd and avoiding the Cd uptake into the cells. Together, IlCDT1 may be a promising gene for the Cd tolerance improvement. SUMMARY: Cysteine-rich gene llCDT1 enhances cadmium tolerance in yeast cells and Arabidopsis thaliana.

De novo characterization of the Iris lactea var. chinensis transcriptome and an analysis of genes under cadmium or lead exposure.

Iris lactea var. chinensis (I. lactea var. chinensis) is tolerant to accumulations of cadmium (Cd) and lead (Pb). In this study, the transcriptome of I. lactea var. chinensis was investigated under Cd or Pb stresses. Using the gene ontology database, 31,974 unigenes were classified into biological process, cellular component and molecular function. In total, 13,132 unigenes were involved in enriched Encyclopedia of Genes and Genomes (KEGG) metabolic pathways, and the expression levels of 5904 unigenes were significantly changed after exposure to Cd or Pb stresses. Of these, 974 were co-up-regulated and 1281 were co-down-regulated under the two stresses. The transcriptome expression profiles of I. lactea var. chinensis under Cd or Pb stresses obtained in this study provided a resource for identifying common mechanisms in the detoxification of different heavy metals. Furthermore, the identified unigenes may be used for the genetic breeding of heavy-metal tolerant plants.

Pb uptake and toxicity to Iris halophila tested on Pb mine tailing materials.

Pb tolerant mechanisms, plant physiological response and Pb sub-cellular localization in the root cells of Iris halophila were studied in sand culture and the Pb mine tailings. Results showed that the activities of superoxide dismutase (SOD) and peroxidase (POD) in the underground parts and the activity of catalase (CAT) in the aboveground and underground parts increased as Pb level was enhanced. Glutathione (GSH) and ascorbic acid (AsA) contents increased by Pb treatments. Pb deposits were found in the middle cell walls or along the inner side of epibiotic protoplasm of some cells which accumulated a large quantity of Pb and died. The dry weights (DWs) of aboveground parts under all Pb tailings treatments decreased insignificantly, while the DW of the underground parts growing in the pure Pb tailings decreased significantly. Pb, Cu, Cd, and Zn contents increased significantly as the levels of Pb tailings were enhanced and Pb contents in the aboveground and underground parts reached 64.75 and 751.75 µg/g DW, respectively, at pure Pb tailings treatment. The results indicated that I. halophila is a promising plant in the phytoremediation of Pb contaminated environment. Some antioxidant enzymes, antioxidants and compartmentalization of Pb were played major roles in Pb tolerance of I. halophila.

Dibutyl Phthalate Inhibits the Effects of Follicle-Stimulating Hormone on Rat Granulosa Cells Through Down-Regulation of Follicle-Stimulating Hormone Receptor.

Dibutyl phthalate (DBP) is used worldwide in solvents and plasticizers. The cytotoxicity and potential tumorigenic effect of DBP have been reported. DBP has also been shown to impact reproductive function. In this study, to further evaluate the effects of DBP on granulosa cells (GCs), we treated rat GCs in vitro with DBP before evaluation of the biological alterations of these GCs. We found that DBP did not induce significant GC death at the tested concentrations. However, follicle-stimulating hormone (FSH)-induced KIT ligand (KITLG) expression in GCs was significantly reduced at both mRNA and protein levels by DBP treatment in a dose-dependent manner. The down-regulation of KITLG was due to the down-regulation of expression of FSH receptor (FSHR) in GCs. Down-regulation of FSHR impaired FSH-induced intracellular signaling in GCs, demonstrated by decreased phosphorylation of AKT and mechanistic target of rapamycin (mTOR). Furthermore, DBP treatment also reduced FSH-induced expression of hypoxia-inducible factor 1-alpha (HIF1A), which is an important signaling component for KITLG expression. Other FSH-induced biological effects, such as production of estradiol and progesterone, as well as GC proliferation, were also suppressed by DBP. Therefore, our study discovered a unique mechanism underlying the toxicity of DBP on GCs. These findings may initiate the development of novel therapeutic interventions for DBP-induced damage to GCs.

Effects of exogenous glutathione and cysteine on growth, lead accumulation, and tolerance of Iris lactea var. chinensis.

Effects of exogenous reduced glutathione (GSH) and cysteine (Cys) on growth, lead (Pb) accumulation, and nonprotein thiol (NPT) contents of Iris lactea var. chinensis under 100 and 500 mg L(-1) Pb stress were studied. Our results showed that 500 mg L(-1) Pb stress caused a dramatical decline in fresh weights, while the reduction of aboveground biomass was alleviated by exogenous GSH and Cys even though keeping higher Pb contents in roots and shoots. Exogenous GSH and Cys could enhance Pb accumulation in the shoots and roots compared with single Pb treatment. The promoting effect of GSH to Pb accumulation was larger than the effect of Cys, and the Pb contents in the shoots and roots treated with 500 mg L(-1) Pb + GSH reached 1,712 and 14,603 mg kg(-1), about 4.19 and 2.78 times of single 500 mg L(-1) Pb treatment, respectively. Microscopic imaging of Pb in roots and leaves showed that higher intensive fluorescence was observed in cell wall of root epidermis, stele, vascular tissues of the roots, and sclerenchyma cells of leaves treated with 500 mg L(-1) Pb + GSH and treated with 500 mg L(-1) Pb + Cys. Exogenous GSH had an apparent promoting effect on root and shoot GSH synthesis, while exogenous Cys reduced the synthesis of cellular GSH in shoot and increased Cys contents. Pb only induced the synthesis of phytochelatin (PC)2 in roots, and the PC2 content declined in GSH- and Cys-treated plant roots. These results suggested that GSH synthesis was a more effective approach to improve Pb accumulation and translocation of I. lactea var. chinensis. Further analysis of protein expression in plants by exogenous GSH and buthionine sulfoximine (BSO) application showed that the proteins regulated by GSH and BSO may constitute various enzymes involved in GSH biosynthesis and play certain roles in Pb accumulation and tolerance of I. lactea var. chinensis.

Reference gene selection for quantitative real-time RT-PCR normalization in Iris. lactea var. chinensis roots under cadmium, lead, and salt stress conditions.

Quantitative real time PCR (RT-qPCR) has emerged as an accurate and sensitive method to measure the gene expression. However, obtaining reliable result depends on the selection of reference genes which normalize differences among samples. In this study, we assessed the expression stability of seven reference genes, namely, ubiquitin-protein ligase UBC9 (UBC), tubulin alpha-5 (TUBLIN), eukaryotic translation initiation factor (EIF-5A), translation elongation factor EF1A (EF1 α ), translation elongation factor EF1B (EF1b), actin11 (ACTIN), and histone H3 (HIS), in Iris. lactea var. chinensis (I. lactea var. chinensis) root when the plants were subjected to cadmium (Cd), lead (Pb), and salt stress conditions. All seven reference genes showed a relatively wide range of threshold cycles (C t ) values in different samples. GeNorm and NormFinder algorithms were used to assess the suitable reference genes. The results from the two software units showed that EIF-5A and UBC were the most stable reference genes across all of the tested samples, while TUBLIN was unsuitable as internal controls. I. lactea var. chinensis is tolerant to Cd, Pb, and salt. Our results will benefit future research on gene expression in response to the three abiotic stresses.

Overexpression of Iris. lactea var. chinensis metallothionein llMT2a enhances cadmium tolerance in Arabidopsis thaliana.

Metallothioneins (MTs) are cysteine-rich, low molecular weight, heavy metal-binding protein molecules. Here, a full-length cDNA homologue of MT2a (type 2 metallothionein) was isolated from the cadmium-tolerant species Iris. lactea var. chinensis (I. lactea var. chinensis). Expression of IlMT2a in I. lactea var. chinensis roots and leaves was up-regulated in response to cadmium stress. When the gene was constitutively expressed in Arabidopsis thaliana (A. thaliana), root length of transgenic lines was longer than that of wild-type under 50µM or 100µM cadmium stress. However, there was no difference of cadmium absorption between wild-type and trangenic lines. Histochemical staining by 3,3-diaminobenzidine (DAB) and nitroblue tetrazoliu (NBT) clearly demonstrated that transgenic lines accumulated remarkably less H2O2 and O2(-) than wild-type. Together, IlMT2a may be a promising gene for the cadmium tolerance improvement.

The physiological response and sub-cellular localization of lead and cadmium in Iris pseudacorus L.

The seedling development and physiological responses of Iris pseudacorus L. to Pb and Cd and their combination were studied for 28 days liquid culture and sub-cellular localization of Pb and Cd in the root tip cells treated with 2,070 mg L(-1) Pb and 1,000 mg L(-1)Cd for 16 days sand culture was evaluated. Results showed that the dry weights (DWs) of shoots and roots of I. pseudacorus were significantly decreased at 500 mg L(-1)Pb and 25 mg L(-1)Cd + 500 mg L(-1)Pb treatments and the root DWs under all treatments were significantly decreased in comparison with that of control. The concentrations of Chla in the leaves were decreased at all treatments, while, the concentrations of Chlb and total carotenoids were not significantly decreased under 25 mg L(-1)Cd and 25 mg L(-1)Cd + 500 mg L(-1)Pb treatments. The MDA and proline concentrations and POD activities in the shoots and roots were increased under treatments of 500 mg L(-1)Pb and 25 mg L(-1)Cd + 500 mg L(-1)Pb, but POD activities in the shoots and roots and MDA concentrations in the shoots were significantly decreased at 25 mg L(-1) Cd treatment. The results of sub-cellular localization of Pb and Cd showed that numerous Pb deposits were found on the inner surface of died cell walls in the cortex treated with 2,070 mg L(-1) Pb and Cd deposits were found in the cell wall treated with 1,000 mg L(-1) Cd. Pb and Cd deposits were not found in the cytoplasm. The results indicated that POD and proline showed strong beneficial properties against Pb and Cd stress and there were some mechanisms keeping most cells with normal activities in the plant from Pb toxicity by sacrificing a few cells that accumulated a large amount Pb. Sub-cellular localizations of Pb and Cd in the root tip cells of I. pseudacorus were little difference with the localizations in other species of Iris in the previous studies.

Cadmium tolerance and accumulation by two species of Iris.

Seedlings of Iris lactea var. chinensis (Fisch.) Koidz. and I. tectorum Maxim. were subjected to 0-160 mg l(-1) Cd in hydroponic system and harvested after 42 days to determine effects on root and shoot dry mass. A subset of 16-day-old seedlings was exposed to 1000 mg l(-1) Cd to characterize sub-cellular localization of Cd in root cells. The Cd contents in the shoots of I. lactea var. chinensis reached 529 microg g(-1 )dry weight (dw) at 80 mg l(-1) Cd treatment and in the shoots of I. tectorum reached 232 microg g(-1) dw at 40 mg l(-1) Cd treatment, without showing signs of visible toxicity. The Cd contents in the shoots of both two test species exceeded 100 microg g(-1), the critical value of Cd hyperaccumulator. The indices of tolerance (ITs) of I. lactea var. chinensis were higher than those of I. tectorum under 10-160 mg l(-1)Cd stress. Sub-cellular localization of Cd in root cells was evaluated using transmission electron microscopy (TEM) and Cd deposits were found in the cell walls, in the cytoplasm and on the inner surface of xylem vessels in the root tip of I. lactea var. chinensis and I. tectorum. A few cells in the root tip of I. tectorum were necrotic. The results showed that the tolerance and accumulation of Cd by I. lactea var. chinensis were higher than those of I. tectorum, suggesting that I. lactea var. chinensis has potential application in phytoremediation.