OBJECTIVETo evaluate the endocrine disrupting effects of cadmium (Cd) using OECD enhanced TG407 test guideline.

METHODSSprague-Dawley (SD) rats were randomly divided into six groups and accordingly administered with 0, 1, 2.5, 5, 10, 20 mg/kg•BW/day of Cd by gavage for 28 days. Body weight, food consumption, hematology, biochemistry, sex hormone levels, urinary β2-microglobulin, organ weights and histopathology and estrous cycle were detected.

RESULTSCd could significantly decrease animals' body weight (P<0.05). Serum luteinizing hormone (LH) at 10-20 mg/kg•BW groups and testosterone (T) at 2.5 and 10 mg/kg•BW groups decreased significantly (P<0.05). However, no statistically significant change was found in urinary β2-microglobulin among Cd-treatment groups (P>0.05). Endpoints related to female reproduction including uterus weight and histopathological change at 10-20 mg/kg•BW groups showed significant increase (P<0.05). While among male rats in 2.5, 10, 20 mg/kg•BW groups, weight of prostate, thyroids, and seminal vesicle glands significantly decreased (P<0.05). Moreover, no histopathological change was observed in kidney.

CONCLUSIONResults suggested that Cd can cause endocrine disrupting effects in SD rats. Comparing with possible renal toxicity of Cd, its toxicity on endocrine system was more sensitive.

Animals ; Body Weight ; drug effects ; Cadmium ; toxicity ; Eating ; drug effects ; Endocrine Disruptors ; toxicity ; Female ; Hormones ; blood ; Kidney ; drug effects ; Male ; Organisation for Economic Co-Operation and Development ; Random Allocation ; Rats, Sprague-Dawley ; Uterus ; drug effects ; beta 2-Microglobulin ; urine

OBJECTIVETo study the expression and localization of co-stimulators in the mucosa of patients with ulcerative colitis (UC), and to explore its role in the pathogenesis of UC.

METHODSExpression of co-stimulators CD86 and inducible co-stimulator (ICOS) was studied by immunohistochemistry on paraffin-embedded mucosal tissue from patients with active UC (64 cases), inactive UC (51 cases) and normal controls (20 cases). Immunostaining for CD28 was also carried out on frozen fresh mucosal tissue sampled from patients with active UC (7 cases), inactive UC (2 cases) and normal controls (5 cases). In addition, expression of CD4, CD8 and CD20 were also examined.

RESULTSIn active UC, increased expression of CD86 was not only observed in lamina propria mononuclear cells but also in the intestinal epithelial cells, as compared with inactive UC and the normal controls (P < 0.01). Increased ICOS expression in lamina propria mononuclear cells was detected in active UC, as compared with inactive UC and the normal controls (P < 0.01). Increased ICOS expression in intestinal epithelial cells was also seen in active UC, as compared with that of inactive UC (P < 0.01). The expression of CD86 was higher in inactive UC than in the normal controls (P < 0.05 or P < 0.01). However, the expression of ICOS showed no statistically significant difference between inactive UC and normal controls. Increased expression of CD28 in active UC, compared with that in inactive UC and normal controls, was also noticed (P < 0.05 or P < 0.01). The number of CD4 or CD8-positive intraepithelial lymphocytes and lymphocytes infiltrating in the lamina propria and small vessel walls was much higher in active UC than in inactive UC and normal controls (P < 0.01). Moreover, the ratio of CD4/CD8 was highest in active UC (P < 0.01). The number of CD20-positive B lymphocytes in lamina propria was also higher in active UC than in inactive UC and normal controls (P < 0.01).

CONCLUSIONSIn active UC, CD86 and ICOS were over-expressed in the intestinal epithelial cells and lamina propria mononuclear cells. The phenomenon suggests that abnormal expression of co-stimulators may contribute to the deregulation of acquired immune responses in UC.

Adult ; Aged ; Antigens, Differentiation, T-Lymphocyte ; metabolism ; B7-2 Antigen ; metabolism ; CD28 Antigens ; metabolism ; CD4-CD8 Ratio ; Case-Control Studies ; Colitis, Ulcerative ; metabolism ; pathology ; Epithelial Cells ; metabolism ; pathology ; Female ; Humans ; Immunohistochemistry ; Inducible T-Cell Co-Stimulator Protein ; Intestinal Mucosa ; metabolism ; pathology ; Leukocytes, Mononuclear ; metabolism ; pathology ; Male ; Middle Aged ; Mucous Membrane ; metabolism ; pathology ; Young Adult

BACKGROUNDNF-kappaB p65 was shown to inhibit transcription of phosphoenolpyruvate carboxykinase (PEPCK), a rate-limiting enzyme in gluconeogenesis in the liver. To understand the mechanism of action of NF-kappaB p65, we investigated the nuclear receptor corepressor in the regulation of PEPCK transcription.

METHODSRat H4IIE cells, human hepatoma HepG2 cells and human embryo kidney (HEK) 293 cells were used in this study. The transcriptional activity of a rat PEPCK gene promoter (-490/+100) was analyzed in HepG2 cells, a HepG2 super suppressor IkBalpha (ssIkBalpha) stable cell line, and HEK 293 cells. The effects of p65 and ssIkBalpha on a rat PEPCK gene promoter were observed using the PEPCK luciferase reporter system. The interaction of the cAMP-response- element-binding (CREB) protein, histone deacetylase 3 (HDAC3) and silencing mediator for retinoic and thyroid hormone receptors (SMRT) with the PEPCK gene promoter were investigated using the chromatin immunoprecipitation (ChIP) assay. p65 cotransfection and RNAi-mediated gene knockdown were used to determine the corepressor involved in the inhibition of PEPCK by NF-kappaB p65 and the transcriptional regulation of CREB by NF-kappaB p65.

RESULTSNF-kappaB p65 inhibited PEPCK expression and the inhibition was blocked by ssIkBalpha. The inhibitory effect of p65 was completely blocked in a HepG2 stable cell line in which ssIkBalpha was expressed. HDAC3 or SMRT knockdown led to a significant up-regulation of PEPCK reporter activity in the presence of p65 cotransfection. In the ChIP assay the interaction of HDAC3 and SMRT with the PEPCK gene promoter was induced by p65 activation, but the CREB signal was reduced. Transcriptional activity of CREB was inhibited by NF-kappaB p65 cotransfection. The inhibitory effect of NF-kappaB p65 was blocked by HDAC3 RNAi or SMRT RNAi.

CONCLUSIONSThe study showed that the inhibition of PEPCK by NF-kappaB p65 was dependent on HDAC3 and SMRT, which form a nuclear corepressor complex for transcriptional inhibition. The transcription factors NF-kappaB p65 and CREB share the same corepressor HDAC3-SMRT, and the corepressor exchange leads to inhibition of PEPCK gene transcription by NF-kappaB p65.

Animals ; Blotting, Western ; Cell Line ; Chromatin Immunoprecipitation ; Cyclic AMP Response Element-Binding Protein ; genetics ; metabolism ; Hep G2 Cells ; Histone Deacetylases ; genetics ; metabolism ; Humans ; NF-kappa B ; genetics ; metabolism ; Nuclear Receptor Co-Repressor 2 ; genetics ; metabolism ; Phosphoenolpyruvate Carboxykinase (ATP) ; genetics ; Promoter Regions, Genetic ; genetics ; Protein Binding ; genetics ; physiology ; Rats ; Reverse Transcriptase Polymerase Chain Reaction ; Transcription Factor RelA ; genetics ; metabolism

BACKGROUNDEstrogen receptor alpha (ER alpha) is the most important endocrine therapy responsiveness predictor for women with breast cancer. The accuracy of the prediction of the response to endocrine therapy was thought to be affected by involving the estrogen receptor coregulatory proteins and cross-talk between ER and other growth factor-signaling networks. Nuclear corepressor 1 (NCOR1) is one of the ER a transcription repressor. The objective of the study is to investigate the expression of NCOR1 at the protein level and pursue its predictive value for breast cancer endocrine therapy.

METHODSIn the present study, the level of expression of NCOR1 protein has been assessed by immunohistochemistry in 104 cases of invasive carcinoma of the breast. Associations between NCOR1 protein expression and different clinicopathological factors and survival were sought.

RESULTSIt was found that NCOR1 was expressed at significantly higher levels in responsive patients treated with endocrine therapy as first-line treatment on relapse. Responsive patients also had a significantly longer post-relapse survival and overall survival. No NCOR1 expression difference was found between patient by age, tumor size, lymph node status, different histological grade groups and human epidermal growth factor receptor 2 (HER2) status. Multivariate analysis showed that NCOR1 is an independent prognostic factor for over-all survival.

CONCLUSIONSIn breast cancer, NCOR1 protein expression level predicts response to endocrine therapy as first-line treatment for breast cancer patients on relapse and NCOR1 protein level assay may increase the accuracy in the endocrine treatment determination and, therefore, improving the patients survival.

Antineoplastic Agents, Hormonal ; therapeutic use ; Breast Neoplasms ; drug therapy ; metabolism ; Estrogen Receptor alpha ; metabolism ; Female ; Gene Expression Regulation ; Humans ; Immunohistochemistry ; Middle Aged ; Nuclear Receptor Co-Repressor 1 ; metabolism ; Receptor, ErbB-2 ; metabolism ; Receptors, Progesterone ; metabolism ; Tamoxifen ; therapeutic use

OBJECTIVETo observe the expression of ER alpha and SMRT in ER alpha-positive and -negative cell lines before and after treatment with tamoxifen (TAM).

METHODSBreast cancer T47D cells (ER alpha-positive) and MDA-MB-231 cells (ER alpha-negative) were treated with TAM, cell viability was measured by MMT assay before and after TAM treatment. Flow cytometry (FCM) was applied to analyze apoptosis rate and cell cycle. Immunohistochemistry and Western blot were used to test ER alpha and SMRT expression in T-47D and MDA-MB-231 cells with and without TAM treatment.

RESULTProliferation rate of T-47D and MDA-MB-231 decreased after 0.10 mmol/L TAM treatment for 48 h compared with control group (P <0.05), especially that of T47D cells. The result of FCM showed that sub-diploid apoptosis peak was found in both cell lines after TAM treatment. Immunohistochemistry and Western blot indicated that T-47D cells presented ER alpha++ and SMRT++, and ER alpha expression decreased after TAM treatment, meanwhile, that of SMRT increased. MDA-MB-231 cells presented ER alpha-, SMRT-, and both expression levels increased slightly after TAM treatment.

CONCLUSIONTAM can inhibit the proliferation of breast cancer cells by inducing cell apoptosis,which is associated with alteration of ER alpha and SMRT expression.

Antineoplastic Agents, Hormonal ; pharmacology ; Apoptosis ; drug effects ; Breast Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Estrogen Receptor alpha ; metabolism ; Female ; Humans ; Nuclear Receptor Co-Repressor 2 ; metabolism ; Selective Estrogen Receptor Modulators ; pharmacology ; Tamoxifen ; pharmacology

The adult worm and plerocercoid larva(sparganum) of Diphyllobothrium mansoni and Moniezia expansa employed in this experiment. The adult worms were divided into three portions, i.e. immature, mature and gravid proglottids, and each proglottids were incubated in 50 cc or 250 cc volume of special incubation flasks with incubation medium consisting of 10 cc of 25 cc of Krebs-Ringer phosphate buffer (pH 7.4). The incubation medium was added C(14)-acetate and non-radioactive carrier Na-acetate so as to contain acetate concentration of 50 mg per cent. The worms were allowed to incubate for 5 hours in the Dubnoff metabolic shaking incubator at 38 C. After incubation period, the lactate and pyruvate appearance rate, total CO(2) production tate, the turnover rates were employed as pervious report(Seo et al., 1965). The quantitative analysis of C(14)-acetate utilized by the adult worm and plerocercoid larva of D. mansoni and M. expansa were compared and discussed in this report. According to these data of the experiment, it is impressed that the fatty acid such as acetate may play a role of major part of their metabolism in the adult worm and plerocercoid larva of D. mansoni , whereas minor part of acetate participated in the metabolism by M. expansa.
parasitology ; helminth ; cestoda ; Diphyllobothrium mansoni ; Moniezia expansa ; sparganum ; acetate ; metabolism ; biochemistry ; acetate ; CO(2) ; Krebs Ringer phosphate buffer

The adult trematodes, Fasciola hepatica, Eurytrema pancreaticum and Paramphistomum cervi, employed in this experiment were obtained from the cattle slaughtered at the local abbatoir. The worms selected and washed several times in normal sterilized saline solution. Each about ten of intact F. hepatica, fourty of E. pancreaticum, and twenty of P. cervi were incubated in 50 cc volume of special incubation flasks with incubation medium consisting of 10 cc. of Krebs-Ringer phosphate buffer(pH 7.4) The incubation medium was added C(14)-1-acetate and non-radioactive carrier Na-acetate so as to contain acetate concentration of 50 mg per cent . The worms were allowed to incubate for 5 hours in the Dubnoff metabolic shaking incubator at 38 C. After incubation period, respiratory CO(2) samples from central well of incubation flask were analysed for total CO(2) production rate and their specific activity of respiratory CO(2). The lactate and pyruvate appearance rates were determined by analyzing the lactate and pyruvate concentration in a medium after incubation. The glycogen samples isolated from worms were analyzed for the tissue concentration and their radioactivities in order to determine the turnover rate of glycogen pool. Radioactivities of these series of experiments were counted by an endwindow Geiger-Muller counter as an infinitely thin samples. The quantitative analysis of C(14)-acetate utilized by F. hepatica, E. pancreaticum and P. cervi were compared and discussed in this report. According to these data of the experiment, it is suggested that the fatty acid such as acetate may play a part of their oxidative process into the respiratory CO2 and the synthetic process into glycogen in the above species of trematodes.
parasitology ; helminth ; trematoda ; Fasciola hepatica ; Eurytrema pancreaticum ; Paramphistomum cervi ; acetate ; metabolism ; biochemistry ; CO(2) ; glycogen ; Krebs-Ringer phosphate buffer

The adult worms of cestodes, Moniezia expansa and Diphyllobothrium mansoni employed in this experiment. The worms were divided into three portions, i.e. immature , mature and gravid proglottids, and each proglottids were incubated in a certain incubation period, and the glucose uptake rate, total CO2 production rate, tissue concentration and their radioactivities were employed as previous reports(Rim et al., 1965). The glucose uptake rate by M. expansa was a mean value of 6.46+/-1.23 micromole per hour per gram of wet wt. and the rate by D. mansoni was a mean value of 18.8+/-0.8 micro-mole per hour per gram of wet wt. The higher rates were observed in the mature proglottid of M. expansa and in the immature proglottid of D. mansoni . The total CO(2) production rates by the worms averaged 14.0+/-2.37 micro-mole per hour per gram in M. expansa and 17.51+/-1.54 micro-mole per hour per gram of wet wt. The relative specific activities of respiratory CO(2)(R.S.A CO(2)) averaged 22.2+/-5.15 percent in M. expansa and 54.2+/-2.2 per cent in D. mansoni. In the both worms, the higher values were obtained in the mature proglottids. Therefore, the average value of 8.84+/-2.66 per cent of glucose utilized by M. expansa and 8.23+/-0.50 percent of glucose utilized by D. mansoni from the medium glucose was oxidized into respiratory CO(2). The tissue concentrations of glycogen were a mean of 2.21+/-0.46 percent per gram of wet wt. in M. expansa and 7.56+/-1.24 percent per gram of wet wt. in D. mansoni. The higher concentration of glycogen was observed in the gravid proglottids of M. expansa, however the gravid proglottids of D. mansoni showed lower concentration of glycogen than the other proglottids. The turnover rate of glycogen pool yielded with a mean of 0.04+/-0.01 miligram per hour per gram of wet wt. of M. expansa, whereas a mean of 1.66+/- 0.46 miligram per hour per gram wet wt. of D. mansoni. Therefore, a mean value of 2.58+/-0.93 per cent(R.G.D gly) of glucose utilized by M. expansa and 53.6+/-1.4 percent by D. mansoni was incorproated into the glycogen . These data account for that at least 11.42 per cent of the utilized glucose by M. expansa and 61.83 per cent of the utilized glucose by D. mansoni participated in furnishing the oxidation into respiratory CO2 and the synthetic process into glycogen.
parasitology ; helminth ; Moniezia expansa ; Diphyllobothrium mansoni ; metabolism ; glucose ; glycogen ; CO(2) ; biochemistry

The fowl nematode Ascaridia galli employed in this experiment was obtained from the intestine of domestic fowls at the local market. The worms selected and washed several times in normal sterilized saline solution. Each about thirty of intact worms were incubated in 50 cc volume of special incubation flasks with incubation mixture consisting of 10 cc of Krebs-Ringer phosphate buffer (pH 7.4) to which were added universally labeled C14-glucose and non-radioactive carrier glucose so as to contain concentration of 200 mg per cent. The worms were allowed to incubation for 3 hours in Dubnoff metabolic shaking incubator at 38 C. After incubation period, respiratory CO(2) samples from central well of incubation flask were analysed for total CO(2) production rate and their specific activity of respiratory CO(2). Glycogen samples isolated from worms were analysed for uptake rate was determined by analyzing the difference of the glucose concentration in a medium before and after incubation period . Radioactivities of these series of experiments were counted by an endwindow Geiger-Muller counter as an infinitely thin samples. The quantitative analysis of C(14)-glucose utilized by Ascaridia galli was summarized as the following . The glucose uptake rate by A. galli was a mean value of 1.73+/-0.32 micro-mole per hour per gram of wet wt. and total CO(2) production rate by the worms averaged 8.44+/-1.11 micro-mole per hour per gram of wet wt. The relative specific activity of respiratory CO(2) (R.S.A CO(2)) averaged 2.68+/-0.38 per cent . Thus , a man of 2.68 per cent of total CO(2) production rate was originated from the glucose in the medium, therefore the rate of CO(2) production derived from medium glucose was a mean of 0.23+/-0.03 micro-mole per hour per gram of wet wt. Thus, the average value of 2.58+/-0.55 percent (R.G.D CO(2))of glucose utilized by the worms from the medium glucose was oxidized to respiratory CO2. The tissue concentration of glycogen in A. galli was a mean of 22.59+/-1.18 miligram per gram of wet wt or 2.26+/-0.123 percent per gram, and the turnover rate of glycogen pool yielded with a mean of 0.17+/-0.04 percent per hour or 0.037+/-0.006 miligram per hour per gram of wet wt. Therefore, a mean value of 16.37+/-4.04 per cent (R.G.D gly) of glucose was incorporated to the glycogen. These data account for that at least 18.95 per cent of the utilized glucose by the worms participated in furnishing the oxidation into respiratory CO(2) and the synthetic process into glycogen. According to the above data of the experiment, it is suggested in the metabolic process of glucose by Ascaridia galli that the synthetic process into the glycogen is more active than the oxidative process into the respiratory CO(2).
parasitology ; helmith ; nematoda ; Ascaridia galli ; metabolism ; biochemistry ; glucose ; CO(2) ; radioactivity ; glycogen ; Krebs Rigner phosphate buffer

The trematode Paramphistomum cervi empolyed in this experiment was obtained from the reticulum of cattle slaughtered at the local abbatoir. The worms were selected and washed several times in normal sterilized saline solution. Each about ten of intact worms were incubated in 50 cc volume of special incubation flasks with incubation mixture consisting of 50 cc of Krebs-Ringer phosohate buffer (pH 7.4) to which were added universally labeled C(14)-glucose and non-radioactive carrier glucose concentration of 200 mg per cent. The worms were allowed to incubate for 3 hours in the incubator at 38 C. After incubation period, respiratory CO(2) samples from central wall of incubation flask were analysed for total CO(2) production rate and their specific activity of respiratory CO(2). Glycogen samples isolated from worms were analysed for the tissue concentration and their radioactivities in order to determine the turnover rate of glycogen pool. The glucose uptake rate was determined by analysing the difference of the glucose concentration in a medium before and after incubation period. Radioactivities of these series of experiments were counted by an endwindow Geiger-Muller counter as an infinitely thin samples. The quantitative analysis of C(14)-glucose utilized by Paramphistomum cervi was summerized as the following. The glucose uptake rate by Paramphistomum was a mean value of 2.32+/-0.27 micro-mole/hr/g of wet wt. and total CO(2) production rate by the worms averaged 10.85+/-0.41 micro-mole/hr/g of wet wt. The relative specific activities of respiratory CO(2) averaged 49.72+/-13.20 per cent. Thus, a mean of 49.72 per cent of total CO(2) production rate was originated from the glucose in the medium, therefore the rate of CO(2) production derived from medium glucose was mean of 5.24+/-2.16 micro-mole/hr/g of wet wt. Thus, the average value of 37.46+/-5.28 per cent of glucose utilized by the worms from the medium glucose was oxidized to respiratory CO(2). The tissue concentration of Paraphismum was a mean of 41.56+/-5.82 micro-mole/hr/g of wet wt or 4.16+/-0.72 per cent/g , and the turnover rate of glycogen pool yielded with a mean of 0.12+/-0.014 percent/hr or 0.06+/-0.04 mg/hr/g of wet wt. Therefore, a mean value of 16.75+/-4.84 per cent of glucose was incorporated to the glycogen. These data account for that at least 54.21 per cent of the utilized glucose by the worms participated in furnishing the oxidation into respiratory CO(2) and the synthetic process into glycogen. According to the above data of the experiment, it is suggested in the metabolic process of glucose by the Paramphistomum that the synthetic process into the glycogen is less active than the oxidative process into the resppiratory CO(2).
parasitology-helminth-trematoda ; Paramphistomum cervi ; autoradiography ; biochemistry ; glucose ; metabolism ; CO(2) ; glycogen