A. officinarum Hance - P. cablin (Blanco) Benth drug pair improves oxidative stress, intracellular Ca2+ concentrations and apoptosis by inhibiting the AGE/RAGE axis to ameliorate diabetic gastroparesis: In vitro and in vivo studies.

ETHNOPHARMACOLOGICAL RELEVANCE: Alpinia officinarum Hance is a perennial natural medicine herbivorous plant, has been used in the management of treat stomach pain and diabetes, it is abundantly cultivated in Qiongzhong, Baisha and other places. P. cablin (Blanco) Benth, one of the most important traditional Chinese plants, which plays functions in antioxidant and gastrointestinal regulation, has been extensively planted in Hainan, Guangdong and other regions. AIM OF THE STUDY: In this study, we investigated the role and underlying molecular mechanism of AP on diabetic gastroparesis (DGP) in vitro and in vivo. MATERIALS AND METHODS: In this study, using ultra-high performance liquid chromatography-mass spectrometry/mass spectrometry (UPLC-MS/MS) to identify active compounds in A. officinarum Hance-P. cablin (Blanco) Benth drug pair (AP). Molecular docking were utilized to explore the potential mechanism of AP treatment of DGP. In in vitro assays, gastric smooth muscle cells (GSMCs) were treated with 35 mM glucose to promote apoptosis and construct the DGP model, which was treated with different concentrations of AP. Furthermore, transfection technology was used to overexpress RAGE in GSMCs and elucidate the underlying mechanisms of alleviation of DGP by AP. RESULTS: Using UPLC-MS/MS analysis, nine components of AP were identified. We found that AP effectively blocked the increase in apoptosis, oxidative stress, and intracellular Ca2+ concentrations. For in vivo experiments, mice were fed with a high-fat irregular diet to construct DGP model, and AP was co-administered via oral gavage daily to prevent the development of DGP. Compared with DGP mice, AP significantly decreased fasting blood glucose levels and increased gastric emptying levels. Consistent with in vitro experiments, AP also considerably decreased the increase in oxidative stress in DGP mice. Mechanistically, AP alleviates apoptosis and DGP by decreasing oxidative stress and intracellular Ca2+ concentrations via the inhibition of the AGE/RAGE axis. CONCLUSIONS: Collectively, this study has established that AP can improve DGP, and the mechanism may be related to the inhibition the AGE/RAGE axis to mitigate apoptosis and DGP. To summarize, this study provides a novel supplementary strategy for DGP treatment.

TYR mutation in a Chinese population with oculocutaneous albinism: Molecular characteristics and ophthalmic manifestations.

Oculocutaneous albinism (OCA) is a rare inherited disorder characterized by a partial or complete reduction of melanin biosynthesis that leads to hypopigmentation in the skin, hair and eyes. The OCA1 subtype is caused by mutations in TYR. The purpose of this study was to investigate the genetic and clinical ophthalmic characteristics of TYR mutations in patients with OCA. Herein, 51 probands with a clinical diagnosis of OCA were enrolled. Whole-exome sequencing and comprehensive ophthalmic examinations were performed. Overall, TYR mutations were detected in 37.3% (19/51) in the patients with OCA. Fifteen patients had compound heterozygous variants, and four cases had homozygous variants. Eleven different pathogenic variants in TYR were detected in these 19 patients, with missense, insertion, delins and nonsense in 71.1% (27/38), 15.8% (6/38), 2.6% (1/38), and 10.5% (4/38), respectively. Clinical examinations revealed that 84.2% (16/19) of patients were OCA1A, and 15.8% (3/19) were OCA1B. Most TYR probands (52.6%, 10/19) had moderate vision impairment, 15.8% (3/19) had severe visual impairment, 10.5% (2/19) exhibited blindness, only 5.3% (1/19) had mild visual impairment and 15.8% (3/19) were not available. Photophobia and nystagmus were found in 100% (19/19) of the patients. In addition, grade 4 foveal hypoplasia was detected in 100% (12/12) of the patients. In conclusion: The TYR patients exhibited severe ocular phenotypes: the majority (93.8%, 15/16) of them had a moderate vision impairment or worse, and 100% (12/12) had severe grade 4 foveal hypoplasia. These novel findings could provide insight into the understanding of OCA.

Acute temperature adaptation mechanisms in the native reptile species Eremias argus.

Reptiles are sensitive to temperature changes as ectotherm animals. The climate warming may pose more serious threat to reptiles. Although the behavior effect and reproduction biology have been well studied, little information is available about the adaptation mechanisms of reptiles to temperature stress. In this study, the native Chinese species, Eremias argus were incubated at 15 (cold stress), 25 (control group) and 35 °C (thermal stress) for 24 h. The transcriptome and metabolome technology were applied to investigate the molecular regulation mechanisms of lizards to acute temperature changes. The CIRBP and HSPA8 were hub genes in response to temperature adaptation. The increased expression of PER gene in lizard circadian rhythm is associated with tyrosine metabolism after cold or thermal stress. The poly-unsaturated fatty acids in female lizard liver were significantly increased with up-regulation of FASN and ACACA genes after thermal stress, which proved the disruption of fatty acid biosynthesis pathway in corporation with the altered body weight. The cortisol and testosterone were important steroid hormones in response to temperature changes especially in male lizard liver. The increased CIRBP gene expression in lizard gonads suppressed the KDM6B gene, which regulates the testis development and may induce sex reversal in male lizard after thermal stress. The adaptation responses of lizards to temperature stress may threaten the health status of wild population.

Development of QSAR models for evaluating pesticide toxicity against Skeletonema costatum.

Nowadays, the emergence of pesticides and its application in agriculture greatly improved the crop quality and food production. However, the resulted ecological problem caused by the widespread pesticide residues attracted more and more attention since the pesticides were harmful to most living organisms. Regulatory agencies such as Environmental Protection Agency (EPA) and European Chemicals Agency (ECHA) stipulated that a comprehensive pesticides risk assessment was essential and also underscored the application of computation method in evaluating pesticides. The present study aimed to use the Quantitative Structure-Activity Relationship (QSAR) method to establish models for quantitatively and qualitatively predicting the toxicity of pesticide against Skeletonema costatum. The regression model was developed using the Genetic Algorithm plus Multiple Linear Regression method and the classification model was established based on the Random Forest algorithm, respectively. Various internal and external validation metrics suggested that the obtained regression model was of good fitness (R2=0.722), robustness (QLOO2=0.653) and external predictive ability (QFn2:0.719-0.776, CCC = 0.878). The classification could correctly predict 79.4% of pesticides in the training set and 69.7% in the validation set. The relatively high sensitivity value of the classification model indicated its good performance in identifying high-toxic pesticides. It could be concluded from the selected modelling descriptors that molecular weight and polarizability impacted the toxicity the most. The atom-type E-state descriptors generally contributed negatively to the pesticide toxicity which verified the negative influence of molecular hydrophilicity. Moreover, the lipophilic, carbon-type, charge related descriptors demonstrated the important influence of lipophilicity and polarity on pesticide toxicity. The models presented in this work could be used to pre-evaluate the toxicity of pesticides within the applicability domain, thus focusing resources on the high-toxic pesticides and assessing the environmental risk of pesticides quickly and economically.

QSAR modeling the toxicity of pesticides against Americamysis bahia.

The widespread use of pesticides has received increasing attention in regulatory agencies because their extensive overuse and various adverse effects on all living organisms. Organizations such as EPA and ECHA have published laws that pesticides should be fully evaluated before bring them to market. In the present study, we evaluated the pesticides toxicity using the Quantitative Structural-Activity Relationship (QSAR) method. The models for the single class pesticides (herbicides, insecticides and fungicides) as well as the general class pesticides (the combined dataset plus some microbicides, molluscicides, etc.) were developed using the Genetic Algorithm and Multiple Linear Regression method. The internal and external validation results suggested that all the obtained models were stable and predictive. According to the modeling descriptors, the lipophilic descriptors contributed positively while all the electrotopological state descriptors showed a negative contribution, their presences in every model verified the conspicuous influence of molecular lipophilicity and hydrophilicity on the pesticides toxicity. However, the influence of topological structure descriptors was different and varies with the physiochemical information they encode. Finally, the models presented in this paper would help assess the pesticides toxicity against Americamysis bahia, shorten test time, and reduce the cost of pesticides risk assessment.

Modeling pesticides toxicity to Sheepshead minnow using QSAR.

Nowadays, the environmental risk caused by the widespread use of pesticides and their ubiquitous residuals has received more and more attention in academia and regulatory agencies. Due to the large number of pesticides used in agriculture and their adverse effects on all living organisms and the numerous end-points, it is necessary to employ the in silico tools to quickly highlight hazardous pesticides. In this study, we have evaluated the toxicity of pesticides against Sheepshead minnow with the Quantitative Structure-Activity Relationship (QSAR) approach. The models for the specific-type (insecticides, herbicides and fungicides) as well as the general-type (combing all the specific-type pesticides and some microbicides, nematicides, etc.) pesticides were developed using the Genetic Algorithm and the Multiple Linear Regression method, subsequently validated with various metrics. The validation results suggested that the obtained models were highly robust, externally predictive and characterized by a broad applicability domain. Considering the modeling descriptors, the toxicity of pesticides would increase with the lipophilicity and decrease with the polarity and hydrophilicity. Most electrotopological state descriptors contribute negatively to the toxicity, while the influence of topological structure descriptors mainly depends on the physiochemical information they encode. The models proposed in this paper would be useful in filling the data gaps, prioritizing and then focusing experiments on more hazardous pesticides.

Enrichment of imidacloprid and its metabolites in lizards and its toxic effects on gonads.

Soil contaminants can cause direct harm to lizards due to their regular swallowing of soil particles. As the world's fastest growing insecticide with long half-life in soil, the endocrine disrupting effect of neonicotinoids on lizards deserves more attention. In this report, we assessed the endocrine disrupting effect of imidacloprid on Eremias argus during 28 days of continuous exposure. Among the imidacloprid and its metabolites, only the metabolite 6-chloropyridic acid had a significant accumulation in the gonads and was positively correlated with its blood concentration. Imidacloprid might cause endocrine disrupting effects on lizards in two ways. First, the desnitro metabolites of imidacloprid could accumulate in the brain, inhibited the secretion of gonadotropin-releasing hormone, and ultimately affected the feedback regulation of hypothalamic-pituitary-gonadal related hormones. Secondly, imidacloprid severely inhibited the gene expression of the corresponding enzymes in the gonadal anti-oxidative stress system, causing histological damage to the gonads and ultimately affecting gonadal function. Specifically, exposure to imidacloprid resulted in abnormal arrangement of spermatogenic epithelial epithelium, hyperplasia of epididymal wall, and oligospermia of male lizard. Meanwhile, gene expressions of cyp17, cyp19, and hsd17ß were severely inhibited in the imidacloprid exposure group, consistent with decreased levels of testosterone and estradiol in plasma. Imidacloprid exposure could cause insufficient androgen secretion and less spermatogenesis in male lizards. The risk of imidacloprid exposure to female lizards was not as severe as that of male lizards, but it still inhibited the expression of cyp19 in the ovaries and led to a decrease in the synthesis of estradiol. This study firstly reported the endocrine disruption of imidacloprid to lizards, providing new data for limiting the use of neonicotinoids.

Unraveling the toxic effects of neonicotinoid insecticides on the thyroid endocrine system of lizards.

The widespread use of neonicotinoids has resulted in large residues in the soil, which has a major impact on the lizards that inhabit the soil. Thyroid hormones play an important role in the growth and development of lizards. In this report, we assessed the disrupting effects of thyroid system on lizards after 28 days of continuous exposure to dinotefuran, thiamethoxam, and imidacloprid, respectively. Neonicotinoid insecticides could seriously affect the concentration of T4 in lizard plasma and the conversion of T4 to T3 in the thyroid gland. Specifically, exposure to dinotefuran affected the intake and utilization of iodine in the thyroid gland, resulting in insufficient thyroid function, which in turn lead to thyroid epithelial hyperplasia and follicular volume enlargement by negative feedback. Exposure to thiamethoxam could activate thyroid function, significantly increasing plasma T3 and T4 concentrations and promoting the binding of T3 and thyroid hormone receptors. Imidacloprid exposure could inhibit the secretion of thyroid hormones, leading to down-regulation of thyroid hormone receptors and related phase II metabolic enzyme genes. This study verified that the continuous exposure of neonicotinoids could affect the lizard thyroid endocrine system. The harm of neonicotinoids to reptiles deserved more attention.

Gonadal disruption after single dose exposure of prothioconazole and prothioconazole-desthio in male lizards (Eremias argus).

Prothioconazole (PTC) is a widely used triazole fungicide with low toxicity, and its desulfurization metabolite, prothioconazole-desthio (PTC-d), is reported to have higher reproductive toxicity to mammals. However, little is known about the reproductive toxicity, much less endocrine disrupting effect, of these two chemicals on reptiles. In this study, we investigated the effects of single dose of PTC/PTC-d (100 mg kg-1 body weight) exposure on the pathomorphism of testes and epididymides, serum sex steroid hormones (testosterone and 17ß-estradiol) and transcription of steroidogenic-related genes (STARD, cyp11A, cyp17, cyp19A, 17ß-HSD, 3ß-HSD, AR and ER-α) in gonads of male lizards (Eremias argus). Although structural disorder existed in PTC-d exposure group, severe gonadal disruption, especially suppression of spermatogenesis was only observed in testis after PTC treatment, which consequently led to the lack of spermatozoa in epididymal ducts. Consistent with this result, T/E2 value in PTC exposure was elevated to a significant higher level compared with control and continually increased over time, while T/E2 value in the PTC-d exposure group slightly increased only at 12 h. These results demonstrated a more serious disruption of PTC on male lizard gonads than PTC-d. In addition, the expression of cyp17 gene was inhibited at 6 h, however, was induced at 12 h, and exhibited negative correlations with STARD, cyp11A and 3ß-HSD after PTC exposure at each timepoint. In PTC-d group, the expression of STARD and 3ß-HSD were significantly down-regulated, in contrast, cyp11A and cyp17 were up-regulated, and each gene showed consistent changes over time. For 17ß-HSD, no significance was observed in both treated groups. This study was the first to compare the gonadal disruption of PTC and PTC-d in male lizards and elucidated that these two chemicals influenced the physiological function of male gonads through differential transcriptional modulation.

Accumulation and toxicity of thiamethoxam and its metabolite clothianidin to the gonads of Eremias argus.

The endocrine disrupting effect of pesticides is considered to be an important factor in the decline of reptile populations. The large-scale application of neonicotinoids in the environment poses a potential threat to small farmland lizards Eremias argus. In this study, we evaluated the disruption effects of thiamethoxam and its metabolite clothianidin on the endocrine disruption of Eremias argus during 28 d exposure. Thiamethoxam and clothianidin could accumulate in the testis and ovary. Adequate blood exchange was the main cause of thiamethoxam and clothianidin accumulation in the gonads. The production of clothianidin aggravated the effect of endocrine disruption to lizards. Thiamethoxam/clothianidin exhibited two distinct ways of interfering with the endocrine disruption of the male and female lizards. Thiamethoxam/clothianidin significantly up-regulated the expression of cyp17 and cyp19 genes in the testis, which ultimately led to a significant decrease in testosterone levels and a significant increase in the 17-estradiol concentrations in plasma. The expression of the estrogen receptor gene in the liver was also significantly increased in male lizards. The significant declines in testosterone and prostaglandin D2 levels in the plasma indicated that thiamethoxam and clothianidin could cause androgen deficiency in male lizards. Meanwhile, in female lizards, thiamethoxam/clothianidin increased the expression of hsd17ß gene in the ovary, causing an increase in testosterone levels in the plasma and an up-regulation of androgen receptor expression in the liver. The effects of thiamethoxam and clothianidin on male lizards were more pronounced. This study verified the possible endocrine disrupting effects of neonicotinoids and provided a new perspective for the study of global recession of reptiles.

Distribution, metabolism and hepatotoxicity of neonicotinoids in small farmland lizard and their effects on GH/IGF axis.

The potential endocrine disruption of neonicotinoids poses a significant threat to the survival of small farmland lizards. We systematically evaluated the distribution, metabolism, and toxicity of three neonicotinoids (dinotefuran, thiamethoxam, and imidacloprid) in the Eremias argus during a 35-day oral administration exposure. Lizards could quickly transfer and store neonicotinoids into the scale and eliminated through molting. Dinotefuran was most prone to accumulation in lizard tissues, followed by thiamethoxam, and imidacloprid was generally present in the form of its terminal metabolite 6-chloropyridinyl acid. Exposure to dinotefuran resulted in hepatic oxidative stress damage, decreased plasma growth hormone concentration, and down-regulation of ghr, igf1 and igfbp2 gene expression. These indicated that dinotefuran might have potential growth inhibition toxicity to lizards. Although imidacloprid caused severe liver oxidative stress damage, the effect of imidacloprid on GH/IGF axis was not obvious. Compared to dinotefuran and imidacloprid, thiamethoxam had the least damage to liver and minimal impact on GH/IGF axis. This study verified the possible damage of neonicotinoids to lizard liver and the interference of GH/IGF axis for the first time.

Study on neurotoxicity of dinotefuran, thiamethoxam and imidacloprid against Chinese lizards (Eremias argus).

The neurotoxicity of dinotefuran, thiamethoxam and imidacloprid against Chinese lizards (Eremias argus) were evaluated in acute oral exposure and 28d subchronic exposure. Dinotefuran was not easily metabolized and showed strong persistence in the lizard brain. Thiamethoxam and imidacloprid were rapidly absorbed and excreted in lizards, and were not easily enriched in the lizard brain. Dinotefuran and thiamethoxam could directly increase the concentrations of acetylcholine in the brain and blood by up-regulating the expression of the ach gene, which in turn enhanced the binding of acetylcholine and acetylcholinesterase receptors, eventually causing the release of dopamine. The effect of dinotefuran was more pronounced than thiamethoxam. Clothianidin was a major metabolite of thiamethoxam in the brain and aggravated the neurotoxic effects of thiamethoxam. Imidacloprid desnitro olefin was the only metabolite of imidacloprid that enriched in the brain. The protonation effect of imidacloprid desnitro olefin was stronger than that of the parent imidacloprid, which increased its binding ability to lizard acetylcholinesterase receptors. Competitive inhibition of imidacloprid desnitro olefin and acetylcholine led to the down-regulation of ach gene expression. Although neonicotinoids caused the opening of ligand-gated ion channel through the activation of acetylcholinesterase receptors, the body would alleviate these effects by the inhibition of voltage-dependent channel activity for compensatory mechanisms. This study provided a new perspective on the neotoxic effects of neonicotinoids.

The metabolism distribution and effect of imidacloprid in chinese lizards (Eremias argus) following oral exposure.

Systematically evaluation of the metabolism, distribution and effect of imidacloprid in Chinese lizards (Eremias argus) were carried out following oral exposure. Imidacloprid-olefin-guanidine was prone to accumulate in the brain and caused potential neurotoxicity. Percutaneous and excretory excretions were the primary ways for the elimination of imidacloprid and its metabolites. Liver was the main site for hydroxy reduction and nitro-reduction metabolism of imidacloprid. The metabolism of imidacloprid was a complex process in which many metabolic enzymes participated. Aldehyde oxidase and CYP2C9 were the key enzymes in nitro-reduction process. CYP3A4 dominated the process of hydroxylation and desaturation. The increase in Glutathione S-transferase expression may be related to the removal of imidacloprid, but also related to the oxidative stress reaction that imidacloprid may cause in tissues, especially in the kidney. The findings enrich and supplement the knowledge of the environmental fate of imidacloprid in reptiles.

The metabolism distribution and effect of dinotefuran in Chinese lizards (Eremias argus).

The Chinese lizards (Eremias argus) were used to evaluate the metabolism, distribution and effect of dinotefuran following oral exposed. The HPLC equipped with Q Exactive focus was used for metabolite identification and concentration analysis. After single oral administration, the time-concentration curves of dinotefuran and its metabolites were tissue-dependent. The liver and kidney were the major metabolic organs. Percutaneous and urinary excretions were the main ways for lizards to eliminate dinotefuran, and the urine output was the limiting factor. Nitro-reduction was an important process of the metabolism of dinotefuran that was dominated by aldehyde oxidase, and P450 enzymes were involved. The CYP3A4 and CYP2C19 played a crucial role in the other metabolic pathways of dinotefuran. The mRNA expressions of GST family were severely inhibited in liver, which showed dinotefuran might pose a risk of damaging the oxidative stress system in liver. Prolonged residuals of dinotefuran and its demethylation metabolite might enhance the risk of dinotefuran to brain. The results enrich and supplement the knowledge of the environmental fate of dinotefuran in reptiles.

Metabolism Distribution and Effect of Thiamethoxam after Oral Exposure in Mongolian Racerunner ( Eremias argus).

Systematic evaluation of the metabolism, distribution, and effect of thiamethoxam in Mongolian racerunner ( Eremias argus) was carried out after oral exposure. HPLC equipped with Q Exactive focus was used for identification and concentration analysis of thiamethoxam and its metabolites. Percutaneous and urine excretions were the primary ways for the elimination of thiamethoxam and its metabolites, and the limiting factor was urine output. Demethylated thiamethoxam and clothianidin were the main metabolites of thiamethoxam in lizards. CYP3A4, CYP3A7, and CYP2C9 played a crucial role in the metabolism process. Aldehyde oxidase only dominated the nitro-reduction process of demethylated thiamethoxam and clothianidin. Glutathione S-transferase might be related to the clearance process of thiamethoxam and its metabolites. The findings indicated that thiamethoxam might pose potential carcinogenic and hepatic injury risk to lizards. The results enrich and supplement the knowledge of the environmental fate of thiamethoxam in reptiles.

The body burden and thyroid disruption in lizards (Eremias argus) living in benzoylurea pesticides-contaminated soil.

Dermal exposure is regarded as a potentially significant but understudied route for pesticides uptake in terrestrial reptiles. In this study, a native Chinese lizard was exposed to control, diflubenzuron or flufenoxuron contaminated soil (1.5 mg kg-1) for 35 days. Tissue distribution, liver lesions, thyroid hormone levels and transcription of most target genes were examined. The half-lives of diflubenzuron and flufenoxuron in the soil were 118.9 and 231.8 days, respectively. The accumulation of flufenoxuron in the liver, brain, kidney, heart, plasma and skin (1.4-35.4 mg kg-1) were higher than that of diflubenzuron (0-1.7 mg kg-1) at all time points. The skin permeability factor of flufenoxuron was more than 20-fold greater than that of diflubenzuron at the end of exposure. However, the liver was more vulnerable in the diflubenzuron exposure group. The alterations of triiodothyronine (T3) and thyroxine (T4) level after diflubenzuron or flufenoxuron exposure were accompanied with the changes in the transcription of target genes involved not only in hypothalamus-pituitary-thyroid (HPT) axis (sult, dio2, trα and udp) but also in metabolism system (cyp1a and ahr). These results indicated that flufenoxuron produced greater body burdens to lizards through dermal exposure, whereas both diflubenzuron and flufenoxuron have the potential to disturb metabolism and thyroid endocrine system.

Oral and dermal diflubenzuron exposure causes a hypothalamic-pituitary-thyroid (HPT) axis disturbance in the Mongolian racerunner (Eremias argus).

Diflubenzuron (DFB) is a potential endocrine-disrupting chemical. However, its thyroid endocrine effect on reptiles has not been reported. In this study, immature lizards (Eremias argus) were exposed to 20 mg kg-1 DFB once a week for 42 days through oral or dermal routes. Their body weight, plasma thyroid hormone levels, thyroid gland histology and the transcription of hypothalamic-pituitary-thyroid (HPT) axis-related genes in different tissues were assessed to explore the effects of DFB on the HPT axis of lizards. The body weight decreased significantly only after the dermal exposure to DFB. Triiodothyronine (T3) to thyroxine (T4) ratio in the male plasma also significantly increased after the dermal exposure. After oral exposure, the activity of thyroid gland was positively related to the thyroid hormone levels. Furthermore, the alterations in thyroid hormone levels affected the HPT axis-related gene expression, which was tissue dependent and sexually selected. The thyroid hormone receptor genes (trα and trß) in the brain and thyroid were more sensitive to oral exposure. However, only the dermal treatment affected the trα, trß and type 2 deiodinase (dio2) genes in the male liver. These results suggest that DFB exposure caused sex-specific changes in the thyroid function of lizards, and the dermal treatment may be an important route for the risk assessment of reptiles.

The tissue distribution, metabolism and hepatotoxicity of benzoylurea pesticides in male Eremias argus after a single oral administration.

Benzoylurea pesticides (BPUs) are widely used to control the locust, but the toxicokinetics and hepatotoxicity of BPUs in lizards have not been investigated. In this study, the tissue distribution, metabolism and liver toxicity of diflubenzuron and flufenoxuron were assessed in the Eremias argus following a single oral exposure. Diflubenzuron preferred to accumulate in the fat and brain (>1.0 mg kg-1) and was rapidly eliminate in other tissues. In the liver, 4-chloroaniline was one of diflubenzuron metabolites, although with a concentration less than 0.05% of the accumulated diflubenzuron. No significant difference was observed in the liver histopathology between the control and diflubenzuron exposure group. The expressions of Cyp1a and Ahr gene which control the cell apoptosis were also equal to the control level. After flufenoxuron exposure, biomodal phenomenon was observed in the liver, skin, brain, gonad, kidney, heart and blood circulation was an important route for the flufenoxuron penetration. The concentrations of flufenoxuron in all tissues were greater than 1.0 mg kg-1 at 168 h. The excretion of flufenoxuron in the faeces was 1.5 fold higher than diflubenzuron. The hepatocytes in the flufenoxuron treated group showed vacuolation of cytoplasm and decreased nucleus. In addition, the Cyp1a and Ahr genes were significantly up-regulated in the flufenoxuron exposure group. These results suggested that the higher hepatotoxicity of flufenoxuron may be attributed to the higher residual level in the lizard tissues and the Cyp1a and Ahr genes can serve as biomarkers to assess the liver toxicity.

Enantioselective metabolism of triadimefon and its chiral metabolite triadimenol in lizards.

Chinese lizards (Eremias argus) were exposed to separated R-(-)-triadimefon, S-(+)-triadimefon and racemic triadimefon to evaluate enantioselective accumulation of triadimefon. After single oral administration of R-(-)-triadimefon, S-(+)-triadimefon and racemic triadimefon, the time-concentration curves in different tissues were found to be different. Triadimefon enantiomers crossed the blood-brain barrier and brain is a main target organ. The residues of triadimefon enantiomers in fat were highest after 24h indicating that fat was the main tissue of accumulation. In racemic triadimefon exposure group, the enantiomer fractions of R-(-)-triadimefon in different tissues showed that the differences between R-(-)-triadimefon and S-(+)-triadimefon were significant in absorption and metabolism, but the differences became smaller in exclusion and accumulation. From the results of mathematical models, S-(+)-triadimefon was absorbed and eliminated faster than R-(-)-triadimefon, and R-(-)-triadimefon was easily distributed in the tissues and more easily converted into its metabolites. Furthermore, among the four enantiomers of triadimenol, SR-(-)-triadimenol produced by S-(+)-triadimefon may have the highest fungicidal activity and the strongest biological toxicity, RR-(+)-triadimenol produced by R-(-)-triadimefon was most likely to bioaccumulate in lizard. Identifying toxicological effects and dose-response relationship of SR-(-)-triadimenol and RR-(+)-triadimenol will help fully assess the risk of TF enantiomers use in the future. The results enrich and supplement the knowledge of the environmental fate of triadimefon enantiomers.

Tissue distribution and metabolism of triadimefon and triadimenol enantiomers in Chinese lizards (Eremias argus).

Triadimefon (TF, S-(+)-TF, R-(-)-TF) and its metabolite triadimenol (TN, TN-A1, A2 and TN-B1, B2) are two systemic fungicides and both of them are chiral pharmaceuticals which are widely used in agricultural industry. Many researches focused on the toxicity effects of triadimefon on mammals, while the ecotoxicological data of tiradimefon on reptiles is limited. In order to understand the toxicity mechanism of triadimefon in reptiles, the current study administrated S-(+)-TF or R-(-)-TF traidimefon (50mg/kgbw) to Chinese lizards (Eremias argus) respectively, the absorption, distribution of triadimefon and the formation of triadimenol were analysed at different sampling times. The metabolic pathways were demonstrated through relative gene expression using quantitative real-time PCR reaction. During the experiment time, triadimefon was quickly peaked to the maximum concentration within 12h in liver, brain, kidney, and plasma, eliminated slowly. The biotransformation in kidney was the lowest and fat possessed the worst degradation ability among others. The metabolite, triadimenol was detected in blood in 2h and reached to a plateau at about 12h in most organs (fat excepted), while the process of metabolism is stereoselective. The mainly metabolite in R-(-)-TF treated group was TN-B1, and TN-A2 in S-(+)-TF group which showed the selective metabolism to other species caused by environmental conditions, differences in the animal models and concentration of TF. The related gene expression of cyp1a1, cyp3a1 and hsd11ß mRNA level in lizards showed different metabolic pathways in the liver and brain. Both P450s enzymes and 11ß-hydroxysteroid dehydrogenase participated in metabolic reaction in liver, while no 11ß-hydroxysteroid dehydrogenase pathway observed in brain. This diversity in liver and brain may cause different degradation rate and ecotoxicological effect in different organs.