Determining 5HT7R's Involvement in Modifying the Antihyperalgesic Effects of Electroacupuncture on Rats With Recurrent Migraine.

Electroacupuncture (EA) is widely used in clinical practice to relieve migraine pain. 5-HT7 receptor (5-HT7R) has been reported to play an excitatory role in neuronal systems and regulate hyperalgesic pain and neurogenic inflammation. 5-HT7R could influence phosphorylation of protein kinase A (PKA)- or extracellular signal-regulated kinase1 / 2 (ERK1 / 2)-mediated signaling pathways, which mediate sensitization of nociceptive neurons via interacting with cyclic adenosine monophosphate (cAMP). In this study, we evaluated the role of 5-HT7R in the antihyperalgesic effects of EA and the underlying mechanism through regulation of PKA and ERK1 / 2 in trigeminal ganglion (TG) and trigeminal nucleus caudalis (TNC). Hyperalgesia was induced in rats with dural injection of inflammatory soup (IS) to cause meningeal neurogenic inflammatory pain. Electroacupuncture was applied for 15 min every other day before IS injection. Von Frey filaments, tail-flick, hot-plate, and cold-plated tests were used to evaluate the mechanical and thermal hyperalgesia. Neuronal hyperexcitability in TNC was studied by an electrophysiological technique. The 5-HT7R antagonist (SB269970) or 5-HT7R agonist (AS19) was administered intrathecally before each IS application at 2-day intervals during the 7-day injection protocol. The changes in 5-HT7R and 5-HT7R-associated signaling pathway were examined by real-time polymerase chain reaction (RT-PCR), Western blot, immunofluorescence, and enzyme-linked immunosorbent assay (ELISA) analyses. When compared with IS group, mechanical and thermal pain thresholds of the IS + EA group were significantly increased. Furthermore, EA prevented the enhancement of both spontaneous activity and evoked responses of second-order trigeminovascular neurons in TNC. Remarkable decreases in 5-HT7R mRNA expression and protein levels were detected in the IS + EA group. More importantly, 5-HT7R agonist AS19 impaired the antihyperalgesic effects of EA on p-PKA and p-ERK1 / 2. Injecting 5-HT7R antagonist SB-269970 into the intrathecal space of IS rats mimicked the effects of EA antihyperalgesia and inhibited p-PKA and p-ERK1 / 2. Our findings indicate that 5-HT7R mediates the antihyperalgesic effects of EA on IS-induced migraine pain by regulating PKA and ERK1 / 2 in TG and TNC.

Partial Purine Nucleoside Phosphorylase Deficiency Helps Determine Minimal Activity Required for Immune and Neurological Development.

Introduction: Complete or near complete absence of the purine nucleoside phosphorylase (PNP) enzyme causes a profound T cell immunodeficiency and neurological abnormalities that are often lethal in infancy and early childhood. We hypothesized that patients with partial PNP deficiency, characterized by a late and mild phenotype due to residual PNP enzyme, would provide important information about the minimal PNP activity needed for normal development. Methods: Three siblings with a homozygous PNP gene mutation (c.769C>G, p.His257Asp) resulting in partial PNP deficiency were investigated. PNP activity was semi-quantitively assayed by the conversion of [14C]inosine in hemolysates, mononuclear cells, and lymphoblastoid B cells. PNP protein expression was determined by Western Blotting in lymphoblastoid B cells. DNA repair was quantified by measuring viability of lymphoblastoid B cells following ionizing irradiation. Results: A 21-year-old female was referred for recurrent sino-pulmonary infections while her older male siblings, aged 25- and 28- years, did not suffer from significant infections. Two of the siblings had moderately reduced numbers of T, B, and NK cells, while the other had near normal lymphocyte subset numbers. T cell proliferations were normal in the two siblings tested. Hypogammaglobulinemia was noted in two siblings, including one that required immunoglobulin replacement. All siblings had typical (normal) neurological development. PNP activity in various cells from two patients were 8-11% of the normal level. All siblings had normal blood uric acid and increased PNP substrates in the urine. PNP protein expression in cells from the two patients examined was similar to that observed in cells from healthy controls. The survival of lymphoblastoid B cells from 2 partial PNP-deficient patients after irradiation was similar to that of PNP-proficient cells and markedly higher than the survival of cells from a patient with absent PNP activity or a patient with ataxia telangiectasia. Conclusions: Patients with partial PNP deficiency can present in the third decade of life with mild-moderate immune abnormalities and typical development. Near-normal immunity might be achieved with relatively low PNP activity.

[Research of inflammatory factors and signaling pathways in knee osteoarthritis].

Knee osteoarthritis is the most common type of arthritis, which is manifested by the deformation and degeneration of articular cartilage and the discomfort of patients with joint pain, which affects the quality of life of patients and aggravates the medical burden of society. The pathogenesis of knee osteoarthritis is very complex. This paper reviews the inflammatory factors and signal pathways involved in knee osteoarthritis. It is found that most of the inflammatory factors involved are interleukin, such as IL-1 ß, IL-6, IL-15, IL-17, IL-18, and tumor necrosis factors, such as TNF-α. These inflammatory factors aggravate knee osteoarthritisby activating corresponding pathways and promoting the release of inflammatory mediators. The inflammatory signaling pathways involved in knee osteoarthritis are complex. Notch pathway, Wnt pathway, SDF-1 / CXCR4 pathway, TLRs pathway, MAPKs pathway, hippo Yap pathway, OPG-RANK-RANKL pathway and TGF-ß pathway are all involved in the regulation of knee osteoarthritis, and the pathways related to inflammatory mechanism are mainly MAPKs pathway and TLRs pathway. Different signaling pathways can cause the destruction of articular cartilage, promote the apoptosis of chondrocytes, and finally lead to the further imbalance of homeostasis in the knee joint. At the same time, the activation of signal pathway can promote the release of inflammatory factors, so under the cascade reaction of inflammatory factors and signal pathway, knee osteoarthritis is aggravating.

Effect of Electroacupuncture on Hyperalgesia and Vasoactive Neurotransmitters in a Rat Model of Conscious Recurrent Migraine.

Migraine onset is associated with the abnormal release of vasoactive neurotransmitters from perivascular nerves, and these neurotransmitters are involved in the pathophysiology of migraine. Hyperalgesia is a key feature of migraine, and accumulating evidence indicates that electroacupuncture (EA) at the single acupuncture point (Fengchi [GB20]) is effective in ameliorating hyperalgesia. In clinical practice, multiple acupuncture points are widely used, especially GB20 and Yanglingquan (GB34). However, the role played by vasoactive neurotransmitters in acupuncture antihyperalgesic effect at the single or multiple acupuncture points remains unknown. We aimed to determine whether EA would exert its antihyperalgesic effects by modulating vasoactive neurotransmitter release from the perivascular nerves. Furthermore, we examined whether targeting multiple acupuncture points would be more effective than targeting a single point in reducing hyperalgesia. The mechanical and thermal hyperalgesia were evaluated by measuring the facial and hind-paw mechanical withdrawal thresholds, tail-flick and hot-plate latencies. Plasma concentrations of vasoactive neurotransmitters were determined using rat-specific ELISA kits from jugular vein, including calcitonin gene-related peptide (CGRP), substance P (SP), vasoactive intestinal peptide (VIP), neuropeptide Y (NPY), pituitary adenylate cyclase-activating polypeptide (PACAP), nitric oxide (NO), and endothelin-1 (ET-1). The result suggested that EA significantly ameliorated the mechanical and thermal hyperalgesia, reduced c-Fos levels in the trigeminal ganglion, and attenuated plasma and dural levels of vasoactive neurotransmitters, especially in the multiple acupuncture points group (GB20+GB34). In conclusion, EA exerts antihyperalgesic effect in a rat model of conscious recurrent migraine, possibly via modulation of the vasoactive neurotransmitters. Furthermore, targeting multiple acupuncture points is more effective than targeting a single point in reducing hyperalgesia.

Early Enzyme Replacement Therapy Improves Hearing and Immune Defects in Adenosine Deaminase Deficient-Mice.

Background: Inherited defects in adenosine deaminase (ADA) cause severe immune deficiency, which can be corrected by ADA enzyme replacement therapy (ERT). Additionally, ADA-deficient patients suffer from hearing impairment. We hypothesized that ADA-deficient (-/-) mice also exhibit hearing abnormalities and that ERT from an early age will improve the hearing and immune defects in these mice. Methods: Auditory brainstem evoked responses, organ weights, thymocytes numbers, and subpopulations, lymphocytes in peripheral blood as well as T lymphocytes in spleen were analyzed in ADA-/- and ADA-proficient littermate post-partum (pp). The cochlea was visualized by scanning electron microscopy (SEM). The effects of polyethylene glycol conjugated ADA (PEG-ADA) ERT or 40% oxygen initiated at 7 days pp on the hearing and immune abnormalities were assessed. Results: Markedly abnormal hearing thresholds responses were found in ADA-/- mice at low and medium tone frequencies. SEM demonstrated extensive damage to the cochlear hair cells of ADA-/- mice, which were splayed, short or missing, correlating with the hearing deficits. The hearing defects were not reversed when hypoxia in ADA-/- mice was corrected. Progressive immune abnormalities were detected in ADA-/- mice from 4 days pp, initially affecting the thymus followed by peripheral lymphocytes and T cells in the spleen. ERT initiated at 7 days pp significantly improved the hearing of ADA-/- mice as well as the number of thymocytes and T lymphocytes, although not all normalized. Conclusions: ADA deficiency is associated with hearing deficits and damage to cochlear hair cells. Early initiation of ERT improves the hearing and immune abnormalities.

Gene therapy for primary immune deficiencies: a Canadian perspective.

The use of gene therapy (GT) for the treatment of primary immune deficiencies (PID) including severe combined immune deficiency (SCID) has progressed significantly in the recent years. In particular, long-term studies have shown that adenosine deaminase (ADA) gene delivery into ADA-deficient hematopoietic stem cells that are then transplanted into the patients corrects the abnormal function of the ADA enzyme, which leads to immune reconstitution. In contrast, the outcome was disappointing for patients with X-linked SCID, Wiskott-Aldrich syndrome and chronic granulomatous disease who received GT followed by autologous gene corrected transplantations, as many developed hematological malignancies. The malignancies were attributed to the predilection of the viruses used for gene delivery to integrated at oncogenic areas. The availability of safer and more efficient self-inactivating lentiviruses for gene delivery has reignited the interest in GT for many PID that are now in various stages of pre-clinical studies and clinical trials. Moreover, advances in early diagnosis of PID and gene editing technology coupled with enhanced abilities to generate and manipulate stem cells ex vivo are expected to further contribute to the benefit of GT for PID. Here we review the past, the present and the future of GT for PID, with particular emphasis on the Canadian perspective.

Profile of organochlorine pesticides in soil cores from some hotspot areas of Beijing, China.

For the valid use of urban land and the safety of public health, an extensive contamination survey of organochlorine pesticides from five soil cores was conducted in the highly urbanized areas of Beijing in China. For topsoils in five soil cores, level of ∑HCHs and ∑DDTs varied from 0.174 to 4,783 ng g(-1) and 0.62 to 57,849 ng g(-1), respectively. The profile in topsoil of cores displayed that levels of pollutants in four soil cores from the urban areas with dense population are higher than that from outskirts. The depth characteristics of pollutants indicated that the level of ∑HCHs and ∑DDTs in BY and WU was close to/below the background value of soil in China (50 ng g(-1)), but those in some soil samples from A, B and ZX core, especially A and B, was above the threshold value for the soil safety (1,000 ng g(-1)). In views of the usage history of the land and profiles of contaminants, the source in A and B probably came from the dumped waste chemical materials in the old chemical factories. The study of soil cores sampled revealed the existence of chemical "time bomb" in urban regions of Beijing. Also, the finding indicated the absolute necessity of assessing the soil quality with the unprecedented land use changes in the big city community.

Thyroid disruption by technical decabromodiphenyl ether (DE-83R) at low concentrations in Xenopus laevis.

Decabromodiphenyl ether (decaBDE), as a flame retardant, is widely produced and used. To study the thyroid disruption by technical decaBDE at low concentrations, Xenopus laevis tadpoles were exposed to technical decaBDE mixture DE-83R (1-1000 ng/L) in water from stage 46/47 (free swimming larvae, system of Nieuwkoop and Faber) to stage 62. DE-83R at concentration of 1000 ng/L significantly delayed the time to metamorphosis (presented by forelimb emergence, FLE). Histological examination showed that DE-83R at all tested concentrations caused histological alterations - multilayer follicular epithelial cell and markedly increased follicle size accompanied by partial colloid depletion and increase in the peripheral colloid vacuolation, in thyroid glands. All tested concentrations of DE-83R also induced a down-regulation of thyroid receptor mRNA expression. These results demonstrated that technical decaBDE disrupted the thyroid system in X. laevis tadpoles. Analysis of polybrominated diphenyl ethers (PBDEs) (sum of 39 congeners) in X. laevis indicated that mean concentrations of total PBDEs in X. laevis exposed to 1, 10, 100, 1000 ng/L were 11.0, 128.1, 412.1, 1400.2 ng/g wet weight, respectively. Considering that PBDEs burden ofX. laevis tadpoles was close to PBDEs levels in amphibians as reported in previous studies, our study has raised new concerns for thyroid disruption in amphibians of technical decaBDE at environmentally relevant concentrations.

Determination of environmentally relevant exposure concentrations of polybrominated diphenyl ethers for in vitro toxicological studies.

Toxicological studies at environmentally relevant concentrations are essential for understanding ecotoxic and health risks of pollutants such as polybrominated diphenyl ethers (PBDEs). However, no information is available on what exposure levels of PBDEs in vitro studies are environmentally relevant. We exposed MCF-7, HepG2, H295R and PC12 cells to BDE-47, and measured BDE-47 concentrations in the cells after exposure. We also used the percentile method to summarize literature data on environmental exposure levels of biotic tissues to PBDEs. The exposure concentration that resulted in a BDE-47 burden in cells close to the 90th percentile of PBDEs levels in tissues was assigned as the upper limit for the environmentally relevant concentration. Exposure to 1nM BDE-47 resulted in PBDEs burdens in MCF-7, HepG2 and H295R cells close to the 90th percentile but PBDEs burdens in PC12 cells were higher than the 90th percentile. In consideration of the high exposure levels in PBDE-polluted areas, we concluded that the highest environmentally relevant exposure concentration of PBDEs in culture media should be approximately 10nM for MCF-7, HepG2 and H295R cells, and<10nM for PC12 cells. These results provide an approximate reference for setting environmentally relevant exposure concentrations of PBDEs for studies in vitro.

Profiles of organochlorine pesticides in earthworms from urban leisure areas of Beijing, China.

In this study, organochlorine pesticides (HCHs and DDTs) in earthworm and soil contacted closely with it were determined for the purpose of the risk assessment of chemicals in the urban leisure environment. The level of total hexachlorocyclohexanes and (HCHs) and dichlorodiphenyltrichloroethanes (DDTs) in earthworms was 0.6500-44.78 ng g(-1) and 18.97-1.112 x 104 ng g(-1), respectively. Absolutely high levels of DDT and its metabolites in earthworm and correlative soils samples, and the bioaccumulation factor (BAF) of DDTs probably presents certain risk to the higher trophic organisms through its food chain, especially birds.

Diffusion of polybrominated diphenyl ether (PBDE) from an e-waste recycling area to the surrounding regions in Southeast China.

Using Cinnamomum camphora (C. camphora) leaves as biomonitors, we investigated that the diffusion of polybrominated diphenyl ethers (PBDEs) from an e-waste recycling area to the surrounding regions. Variance in sigma(32)PBDEs concentrations (0.46-399.93 ng g(-1) dry weight) in the leaves showed that PBDEs from the e-waste recycling area diffused into the surrounding regions, and resulted in a halo of PBDEs contamination, at least 74 km in radius. The attenuation of sigma(32)PBDEs in the diffusing process fitted in log-linear regression. The difference in the attenuating slopes of linear equations among different directions can be explained by terrain and wind direction. The attenuation of PBDE congeners also fitted well in log-linear regression. The findings that similar attenuating slopes and characteristic travel distance among congeners suggest that the transport behavior of lower brominated congeners might not differ from that of higher brominated congeners in short-range scale.

Levels and distribution of organochlorine pesticides in various media in a mega-city, China.

As one of China's great metropolises, Taiyuan is generally recognized to be one of the most polluted cities in the world. It is affected by pollution from heavy industries, and from organochlorine pesticides (OCPs). In this study, we measured the concentrations of OCPs in various environmental media in Taiyuan. We measured the levels of OCPs in 15 soil samples and 34 samples of respirable particulate matter (PM) (17 of PM(2.5) and 17 of PM(10)) from urban areas of Taiyuan. Analysis of the composition ratios of OCP residues revealed that dichlorodiphenyltrichloroethane (DDT) and hexachlorocyclohexane (HCH) were the predominant compounds in soil, PM(2.5) and PM(10). The total DDT concentrations were 1.8-1.0x10(2)ngg(-1) in soil, 6.5-1.0x10(3)pgm(-3) in PM(2.5), and 8.5-1.5x10(3)pgm(-3) in PM(10). The total HCH concentrations were 1.4-45ngg(-1) in soil, 5.2-8.0x10(2)pgm(-3) in PM(2.5), and 9.6-8.7x10(2)pgm(-3) in PM(10). Furthermore, source analysis indicated that there was new input of OCPs at some sites. Finally, analysis of the individual OCPs revealed that new OCP contamination was the major problem in Taiyuan. This study suggests that measures should be taken to reduce the levels of OCP residues in order to improve the environmental quality in Taiyuan.

Levels and chiral signatures of organochlorine pesticides in urban soils of Yinchuan, China.

In this study, residual level and enantiomeric composition of typical organochlorine pesticides (OCPs) were surveyed in urban soils of Yinchuan, China. The median levels of summation Sigma HCHs and summation Sigma DDTs were 0.852 and 2.24 ng/g, respectively, which suggested little risk for ecological environment and human health in the study area. Both chiral alpha-HCH and o,p'-DDT displayed the non-racemic signatures in all samples. The isomer ratios of summation Sigma HCHs and summation Sigma DDTs combined with enantiomer fractions (EFs) of alpha-HCH and o,p'-DDT, suggested that contamination source of HCHs derived from historical HCHs (including technical HCHs and Lindane) and that of DDTs originated from old source with the usage of mixed technical DDTs and dicofol.

Organochlorine pesticides in particulate matter of Beijing, China.

Eighteen organochlorine pesticides (OCPs) in air particulate matter (PM) samples collected from 20 sites in Beijing, China in 2005 and 2006 were analyzed to determine the composition, spatial distribution and sources. Total OCPs of PM2.5 and PM10 ranged from 32.59 pg m(-3) to 9232.31 pg m(-3) and 80.96 pg m(-3) to 9434.97 pg m(-3), respectively, categorized as heavier pollution compared to other cities of China. Among six kinds of functional zones involved, commercial areas and industrial centers were heavily polluted. Distinct concentrations variations were detected in PM with different particle diameters. Pollution distribution was spatially heterogeneous inside the city. The most heavily polluted sites located in the southeast part of the city, where laid many chemical factories. Compositional analysis and principal component analysis (PCA) suggested that past usage of OCPs was the main source, though there were new inputs in some sites.

Contamination of soils with organochlorine pesticides in urban parks in Beijing, China.

Urban parks are an integral component of healthy urban living. Since they are frequently visited, an understanding of the environmental quality of these urban facilities is crucial. Here, a study was conducted on the contamination of soils in the parks of Beijing. Organochlorine pesticides (OCPs), which have the potential to cause endocrine disturbances, were considered study objectives. Hexachlorocyclohexanes (HCHs) were found at concentrations of 0.2490-197.0 ng g(-1) and dichlorodiphenyltrichloroethanes (DDTs) were found at concentrations of 5.942-1039 ng g(-1) in the soils investigated. The preliminary pollution assessment indicated that DDTs have caused high pollution levels in the soils of some parks. Analysis of the sources of contamination showed that HCHs in the soils were derived from an old mixed source of technical HCHs and lindane and that DDTs, which were suspected to have recent application to the soils at some sites, were derived mainly from a mixture of technical DDTs and dicofol containing DDT impurities. An independent sample t-test proved that pesticides containing DDTs had been used in large amounts in the soils of parks before 1983 (p<0.05) and that the levels of DDTs in the soils of parks administered by the Beijing municipal government were significantly higher than the levels in those administered by the district government (p<0.05). However, the main difference in this situation needs to be further studied. This study suggested that open spaces like urban parks were not as sound as was expected and that there was potential for exposure of visitors/workers in the parks to organochlorine pesticides.

Residues of organochlorine pesticides in surface soils from college school yards in Beijing, China.

Organochlorine pesticides (OCPs) have been a major environmental issue, attracting much scientific concern because of their nature of toxicity, persistence, and endocrine disrupting effects. Soil samples were collected from ten college school yards in Beijing in 2006 and analyzed to determine fifteen OCPs. Dichlorodiphenyltrichloroethanes (DDTs) were found to be the main pollutants, accounting for 93.70% of total OCPs, followed by hexachlorohexanes (HCHs) (2.25%) and hexachlorobenzene (HCB) (1.82%). Content of chlordanes (CHLs), heptachlors (HEPTs), and endosulfans comprised 0.51%, 1.05%, and 0.79% of fifteen OCPs, respectively. The preliminary pollution assessment indicated that DDTs have caused high OCPs levels in some schools. Source identification showed that HCHs in soils were originated from an old mixed source of technical HCHs and lindane. And DDTs were mainly from mixed use of technical DDTs and dicofol containing DDT impurities. According to GB15618-1995 (guidelines of Chinese environmental quality standards for soils), HCHs and DDTs levels might be categorized as little and low polluting pesticides. This study indicated that the environmental quality of college school yards with large green land were not as good as was expected and there existed potential exposure risk of college population to OCPs.

Feminizing/demasculinizing effects of polychlorinated biphenyls on the secondary sexual development of Xenopus laevis.

We have previously demonstrated that polychlorinated biphenyls (PCBs) have caused phenotypic feminization/demasculinization of gonadal development in Xenopus laevis. Whether PCBs affect secondary sexual development has remained unknown. In this study, X. laevis tadpoles were exposed to Aroclor1254 and PCB(3) from stage 46/47 (system of Nieuwkoop and Faber) for up to 1 month postmetamorphosis. After 24 months postmetamorphosis, the degree of secondary sexual development was examined. Male oviducts were observed in some of the PCB-exposed male frogs, but not in control males. These male oviducts had not completely developed in histological structure when compared with mature female oviducts. Larynx weight and width of PCB-exposed males were significantly less than those of control males. Laryngeal histology showed that PCBs inhibited cartilaginous and muscular development of male frogs, i.e. elastic cartilages had not completely developed and laryngeal muscle fibers were smaller. In a further study on adult male frogs, a decrease in serum testosterone level was found in PCB-exposed frogs compared with controls, but serum estradiol level was not significantly affected. Our study suggests that PCBs can cause phenotypic feminization/demasculinization of male genital ducts and larynges, and these effects may, in part, result from the decrease in serum testosterone level in X. laevis.

Application of liquid-phase microextraction and gas chromatography-mass spectrometry for the determination of chloroform in drinking water.

In this paper, a novel method for the determination of chloroform in drinking water has been described. It is based on liquid-phase microextraction (LPME) and gas chromatography-mass spectrometry (GC-MS). Extraction conditions such as solvent selection, organic solvent dropsize, stirring rate, content of NaCl and extraction time were found to have significant influence on extraction efficiency. The optimized conditions were 1.5 microl xylene, 20 min extraction time at 400 rpm stirring rate without NaCl addition. The linear range was 1.0 - 100 microg l(-1) for chloroform. The limit of detection (LOD) was 1.0 microg l(-1); and relative standard deviation (RSD) at the 30 microg l(-1) level was 2.9%. Tap water samples from a laboratory were successfully analyzed using the proposed method. The relative recovery of spiked water samples was 104%.

A novel headspace solid-phase microextraction method for the exact determination of organochlorine pesticides in environmental soil samples.

A novel method of determining organochlorine pesticides (OCPs) is described. It is based on solid-phase microextraction (SPME) and gas chromatography-electron capture detection. During the development of the method, soil samples were prepared, spiked with standard solution, and then aged for some time. Extraction conditions such as the extraction time, the NaCl content, the volume of water, the extraction temperature and the desorption time were investigated and optimized. The limits of detection obtained using the method ranged from 0.10 to 0.51 ng g(-1), and relative standard deviations were lower than 10% for most organochlorine pesticides. Real soil samples were successfully analyzed using the proposed method. The results from the method developed here were in good agreement with those obtained using ultrasonic extraction. The result demonstrates that aging soils spiked with standard solution is an important method development step, because the soil samples obtained using this approach are more like real soils than those obtained when aging is not used.

Xenopus laevis is a potential alternative model animal species to study reproductive toxicity of phytoestrogens.

This study investigated effects of phytoestrogen quercetin on the gonadal development in Xenopus laevis. X. laevis at Nieuwkoop and Faber stage 46/47 were exposed to 50, 100 and 200 microg/L quercetin till 1 month postmetamorphosis. Gonads from frogs at 1 and 3 months postmetamorphosis were examined in gross morphology and histology. The highest dose of quercetin as well as estradiol (E2) significantly increased the percentages of phenotypic females. Exposure to quercetin at all doses induced abnormal testes with certain ovarian characteristics to some degree in gross morphology, including ovotestes. The abnormality rate exceeded 10% in each quercetin treatment. Histologic examination revealed that some abnormal testes exhibited intersexuality with testicular structure and ovarian structure or oocytes interspersed in testicular structure at 1 month postmetamorphosis. At 3 months postmetamorphosis, testicular abnormalities were more obvious, such as necrosis or apoptosis of spermatogonia, occurrence of developed or undeveloped oocytes, delay of the development of seminiferous tubes without or less late stage spermatocytes. The results have shown that quercetin cannot only feminize but also impair testicular development of X. laevis, i.e. X. laevis is sensitive to phytoestrogen. It is suggested that X. laevis might be an alternative model species to study reproductive toxicity of phytoestrogens.