Analysis of individual and combined estrogenic effects of bisphenol, nonylphenol and diethylstilbestrol in immature rats with mathematical models

BACKGROUND@#Traditional toxicological studies focus on individual compounds. However, this single-compound approach neglects the fact that the mixture exposed to human may act additively or synergistically to induce greater toxicity than the single compounds exposure due to their similarities in the mode of action and targets. Mixture effects can occur even when all mixture components are present at levels that individually do not produce observable effects. So the individual chemical effect thresholds do not necessarily protect against combination effects, an understanding of the rules governing the interactive effects in mixtures is needed. The aim of the study was to test and analyze the individual and combined estrogenic effects of a mixture of three endocrine disrupting chemicals (EDCs), bisphenol A (BPA), nonylphenol (NP) and diethylstilbestrol (DES) in immature rats with mathematical models.@*METHOD@#In the present study, the data of individual estrogenic effects of BPA, NP and DES were obtained in uterotrophic bioassay respectively, the reference points for BPA, NP and DES were derived from the dose-response ralationship by using the traditional no observed adverse effect (NOAEL) or lowest observed adverse effect level (LOAEL) methods, and the benchmark dose (BMD) method. Then LOAEL values and the benchmark dose lower confidence limit (BMDL) of single EDCs as the dose design basis for the study of the combined action pattern. Mixed prediction models, the 3 × 2 factorial design model and the concentration addition (CA) model, were employed to analyze the combined estrogenic effect of the three EDCs.@*RESULTS@#From the dose-response relationship of estrogenic effects of BPA, NP and DES in the model of the prepuberty rats, the BMDL(NOAEL) of the estrogenic effects of BPA, NP and DES were 90(120) mg/kg body weight, 6 mg/kg body weight and 0.10(0.25) μg/kg body weight, and the LOAEL of the the estrogenic effects of three EDCs were 240 mg/kg body weight, 15 mg/kg body weight and 0.50 μg/kg body weight, respectively. At BMDL doses based on the CA concept and the factorial analysis, the mode of combined effects of the three EDCs were dose addition. Mixtures in LOAEL doses, NP and DES combined effects on rat uterine/body weight ratio indicates antagonistic based on the CA concept but additive based on the factorial analysis. Combined effects of other mixtures are all additive by using the two models.@*CONCLUSION@#Our results showed that CA model provide more accurate results than the factorial analysis, the mode of combined effects of the three EDCs were dose addition, except mixtures in LOAEL doses, NP and DES combined effects indicates antagonistic effects based on the CA model but additive based on the factorial analysis. In particular, BPA and NP produced combination effects that are larger than the effect of each mixture component applied separately at BMDL doses, which show that additivity is important in the assessment of chemicals with estrogenic effects. The use of BMDL as point of departure in risk assessment may lead to underestimation of risk, and a more balanced approach should be considered in risk assessment.

Eficacia y seguridad del dietiletilbestrol, en el tratamiento del cáncer de próstata resistente a la castración / Effectiveness and Safety of Diethylstilbestrol in the Treatment of Castration Resistant Prostate Cancer

Introducción y objetivos Existen múltiples opciones de tratamiento en pacientes con cáncer de próstata resistente a la castración, entre ellas el Dietilestilbestrol (DES) sin embargo, su uso es controversial. Este estudio tiene como objetivo determinar la eficacia y seguridad del DES, en el tratamiento de pacientes con cáncer de próstata resistente a la castración en nuestra población. Métodos y Materiales Se realizó un estudio de corte transversal, incluyendo los pacientes con cáncer de próstata resistente a la castración que recibieron tratamiento con DES. Se realizó un análisis demográfico, bivariado, tomando como desenlace la respuesta del PSA (Ausente, Completa o Parcial), el tiempo medio de progresión del PSA y la presencia de eventos adversos asociados al medicamento. Resultados Noventa y un pacientes incluidos al final del estudio. La distribución de respuesta del PSA fue así: Respuesta en 57 (63,7%) pacientes, (Completa 28% - 31,1% y parcial 29% - 32,2%). Ausente en 33 (36,7%) pacientes. El análisis bivariado no evidenció asociación entre las variables y los desenlaces propuestos. El tiempo medio de progresión del PSA fue de 10,43 meses (Log-rank p = 0.001), no se encontraron diferencias estadísticamente significativas para el tiempo medio a progresión en asociación con la respuesta al PSA (respuesta o ausente y la presencia de enfermedad metastásica), Log-rank p = 0,789, Log-rank p = 0,218, Log-rank p = 0,780 respectivamente. La tasa de complicaciones asociadas a DES fue del 4,4% y correspondió en todos los casos a trombosis venosa profunda. Conclusiones El DES en pacientes con cáncer de próstata resistente a la castración continúa siendo una herramienta de tratamiento eficaz y con baja tasa de eventos adversos en nuestra población.

Introduction and Objectives There are multiple treatment options in patients with castration-resistant prostate cancer, including diethylstilbestrol (DES), but its use is controversial. This study aims to determine the efficacy and safety of DES in the treatment of patients with castration resistant prostate cancer in our population. Methods and Materials A cross-sectional study was performed, including patients with castration resistant prostate cancer who were treated with DES. A demographic analysis was performed, bivariate analysis, taking as outcome PSA response (Complete or partial), the median time to PSA progression and the presence of adverse events associated with the drug. Results 91 patients included at the end of the study. The distribution of PSA response was so; Response in 57 (63.7%) patients (Full 28 to 31.1% and partial 29 to 32.2%). Absent in 33 (36.7%) patients. Bivariate analysis evidenced no association between the variables and proposed outcomes. The median time to PSA progression was 10.43 months (log-rank p = 0.001), no statistically significant differences in the average time to progression was found in association with PSA response (response or absent and the presence of disease metastatic), Log-rank p = 0.789, log-rank p = 0.218, log-rank p = 0.780 respectively. The rate of complications associated with DES was 4.4% and corresponded in all cases to deep vein thrombosis. Conclusions DES in patients with resistant prostate cancer castration continued to be an effective choice of tool of treatment with a low rate of adverse events in our population.

Establishment of a rat model for uterine leiomyomas based on Western and traditional Chinese medicine theories

Uterine leiomyomas (ULs) are benign monoclonal tumors that arise from the underlying myometrial tissue in the uterus. Effective therapies are still lacking because of poor understanding of the pathophysiology and epidemiology. Hence, it is urgent to establish efficient animal models to screen novel anti-UL therapies. In this study, for the first time, traditional Chinese medicine and Western medicine were combined to establish an animal model of ULs in rats. In order to evaluate the function and value of the novel model, it was compared with other models. The long-term and short-term rat models for ULs were established using progesterone and diethylstilbestrol. Rats in Qi stagnation and blood stasis group were injected with epinephrine hydrochloride and received chronic unpredictable stress for two weeks. Rats in combining disease with syndrome group (CDWSG) received not only epinephrine hydrochloride injection and chronic unpredictable stress but also progesterone and diethylstilbestrol treatment. We analyzed differences in organ coefficient, uterus size, uterine pathology, concentrations of progesterone, estradiol, progesterone receptor, estrogen receptor, expression of desmin, α-smooth muscle actin, and vimentin among the five groups. The animal model of ULs was successfully constructed by loading the rats with estrogen and progesterone. The rat model of CDWSG was more stable than other groups and the method was the most efficient.

Impacts of DES on the expressions of related genes in the gubernaculums testis of newborn mice / 中华男科学杂志

Objective@#To investigate the influence of diethylstilbestrol (DES) on the mRNA expressions of the androgen receptor (AR), estrogen receptor α (ERα), proliferating cell nuclear antigen (PCNA), and actin alpha 1 (ACTα1) in the gubernaculums testis of newborn mice and explore their action mechanisms.@*METHODS@#A total of 140 male Kunming mice were randomly divided into a blank control, a dimethyl sulfoxide (DMSO) control, and 5 experimental groups to be treated subcutaneously with normal saline, DMSO, and DES at 0.02, 0.1, 0.5, 10 and 50 μg per kg of the body weight per day, respectively, at gestation days 9－17. On the first day after birth, the animals were sacrificed and the gubernaculums testis collected for detection of the mRNA expressions of AR, ERα, PCNA and ACTα1 by RT-PCR.@*RESULTS@#Compared with the DMSO control, the experimental groups, particularly the DES 10 and 50 μg groups, showed significant increases in the mRNA expression of ERα (RE2 = 0.825, P <0.05), but remarkable decreases in those of AR, PCNA and ACTα1 (RA2 = 0.713, RP2 = 0.946, RT2 = 0.960, P <0.01), all in a dose-dependent manner.@*CONCLUSIONS@#The AR, ERα, PCNA, and ACTα1 mRNA are expressed in the gubernaculum testis of normal newborn mice, and their expression levels may be influenced by intervention with different concentrations of DES during the gestation. Exogenous estrogens may affect the proliferation and contraction of gubernaculum testis cells and consequently the normal development of the testis or even the whole male reproductive system by influencing the metabolism of ER and/or AR.

Rapid detection of diethylstilbestrol using a quartz crystal microbalance with gold nanoparticals amplification / 中华预防医学杂志

<p><b>OBJECTIVE</b>To develop a quartz crystal microbalance (QCM) immunosensor with high sensitivity and selectivity for the rapid detection of diethylstilbestrol.</p><p><b>METHODS</b>Dextran was used as reducing agent for preparing gold nanoparticles (AuNPs) with the size of 40 nm. The AuNPs were coupled with anti-DES antibody after amination. A monolayer was generated after immersing the quartz crystal into the solution of 5 mmol/L 11-mercaptoundecanoic acid(MUA) for 16 hours. After the monolayer was activated by 1-ethyl-3-(3-dimethylaminopropry) carbodiimide hydrochloride (EDC·HCl) and N-hydrosuccinimide (NHS), 20 μl of 2.2 mg/ml DES-HS-BSA was dropped onto the surface of crystal to prepare a sensitive membrane which can recognize DES specifically. Then, 50 μl of 1 mol/L ethanolamine (pH 8.5) was used to seal the carboxylic groups to make the sensitive membrane which could identify DES specifically. QCM immunosensor was used as detection platform to optimize the reaction conditions. Under the optimized conditions, 10 μl of 28 μg/ml AuNPs-antibody was mixed with 10 μl of 0.03-2.5 μg/ml DES, and the mixture was added on the sensitive membrane. QCM immunosensor was used to detect the signals and the standard curve was obtained at the same time. The detection limit was calculated based on the standard curve. The specificity was evaluated by testing DES and its analogues with the same concentration.</p><p><b>RESULTS</b>The optimized concentration for the immobilization of DES-HS-BSA on the surface of QCM was 2.2 mg/ml. The optimized concentration for coupling anti-DES antibody with AuNPs was 7 μg/ml and 15 nmol/L, respectively. The optimized concentration of AuNPs-antibody was 14 μg/ml. The logarithm of DES concentration was proportional to the frequency shift in the range of 0.16-500 ng/ml, Δf=-24.170 lgCDES+69.71, R(2)=0.998. The detection limit of this method was 0.13 ng/ml. DES analogues could not influence the detection of DES obviously, so the sensor had good specificity.</p><p><b>CONCLUSION</b>The quartz crystal microbalance immunosensor with gold nanoparticals amplification could detect DES sensitively and rapidly.</p>

Uterine Clear Cell Carcinoma of Postmenopausal Woman: A Case Report

Endometrial cancer is the most common gynecologic malignancy in developed countries. Clear cell carcinoma typically occurs in the ovaries, and very rarely occurs in the endometrium; it accounts for less than 3% of all endometrial cancers. It is presumed that clear cell carcinomas are of Müllerian duct origin, and an association with exposure to diethylstilbestrol (DES) or other nonsteroidal follicle stimulating hormones has been described. We report a case of a postmenopausal woman who presented with vaginal bleeding without a specific medical history. Under the impression of an endometrial mass, we performed a laparoscopic operation. Pathologic results showed clear cell carcinoma of the endometrium. Depth of invasion was 0.2 cm out of a 0.5 cm total thickness, and the rectal shelf mass was clear cell carcinoma. We report the case with a brief review of the relevant literature.

Expression of nNOS and ultrastructural changes in the penile tissue of rats with prolactinoma-induced erectile dysfunction / 中华男科学杂志

<p><b>OBJECTIVE</b>To study the expression of nNOS and ultrastructural changes in the penile tissue of rats with prolactinoma-induced erectile dysfunction (ED).</p><p><b>METHODS</b>We established the model of prolactinoma in 20 male Westar rats by peritoneal injection of diethylstilbestrol (DES) and treated the control rats with normal saline (n = 10) or sterilized arachis oil (n = 10). After 8 weeks, we performed the apomorphine test and measured the weight of the pituitary gland and the levels of serum prolactin (PRL) and testosterone (T) to confirm the successful construction of the prolactinoma-induced ED model. Then we determined the expression of nNOS in the penile tissue by immunohistochemistry and examined the ultrastructural changes of the penile cavernosum under the transmission electron microscope.</p><p><b>RESULTS</b>The prolactinoma-induced ED model was successfully established in 15 rats. The weight of the pituitary gland was significantly increased in the rats treated with DES as compared with the normal saline and sterilized arachis oil controls ([46.7 ± 15.5] vs [11.7 ± 2.4] and [12.4 ± 2.3] mg, both P < 0.05). The level of serum PRL was markedly higher while that of T remarkably lower in the former than in the latter two groups ([1,744.9 ± 304.5] vs [11.5 ± 2.4] and [10.6 ± 1.9] ng/ml, both P < 0.0l; [1.54 ± 0.46] vs [3.11 ± 1.08] and [3.04 ± 1.11] ng/ml, both P < 0.05). The rate of penile erection was significantly reduced in the prolactinoma-induced ED model rats in comparison with the normal saline and arachis oil controls (16.7% vs 100% and 87.5%, both P < 0.05), and so was the expression of nNOS in the penile tissue (0.024 ± 0.011 vs 0.066 ± 0.019 and 0.058 ± 0.021, both P < 0.05). Transmission electron microscopy manifested significant ultrastructural changes in the endothelial and smooth muscle cells of the cavernous tissue in the prolactinoma-induced ED models.</p><p><b>CONCLUSION</b>The ultrastructural changes of the penile cavernous tissue and the reduced expression of nNOS in penile tissue may be the most important mechanisms of prolactinoma-induced ED in rats.</p>

Tratamiento con dietilestilbestrol en pacientes refractarios a la castración por análogos LH- RH / Treatment with diethylstilbestrol in patients who had failed to androgen deprivation by lhrh agonists

Introducción: El objeto de este trabajo clínico es estudiar la eficacia de la administración de Dietilestilbestrol (DES) oral en pacientes con cáncer de próstata avanzado y que han presentado refractariedad al tratamiento con análogos LH-RH y además evaluar los efectos colaterales atribuibles a su uso. Material y métodos: Entre Noviembre de 2010 y Mayo de 2012 se ingresaron en forma consecutiva al estudio 15 pacientes con cáncer prostático avanzado, refractarios a manejo con análogos LH-RH. Edad promedio de los pacientes 69,4 años .Rango 57-80. Se registró el tipo de tratamiento realizado, detallando el manejo hormonal previo al que fueron sometidos. Se consideró refractariedad a los análogos, la detección de 2 alzas consecutivas del APE durante la administración de éstos. Se indicó a los pacientes 1mg. de DES al día. La evaluación del tratamiento se hizo cada 30 días con determinación de APE y registro de efectos colaterales. Resultados: De los 15 pacientes, 8 (53,3 por ciento) disminuyeron su APE inicial, 6 de ellos (40 por ciento) lo hicieron en más de un 50 por ciento de su valor y 2 (13,3 por ciento) disminuyeron el APE pero en menos de un 50 por ciento. 7 pacientes (46,6 por ciento) no disminuyeron su valor de APE y fueron considerados como fracasos. Como efectos colaterales del tratamiento tuvimos 13 pacientes (86,6 por ciento) que presentaron hipersensibilidad de los pezones, pero solo 2 requirieron tratamiento sintomático. 4 pacientes (26,6 por ciento) desarrollaron ginecomastia leve a moderada y no tuvimos ninguna complicación cardiovascular en los 15 pacientes estudiados. Conclusión: Consideramos de acuerdo a nuestros resultados que el DES oral es efectivo como tratamiento en los pacientes que han fallado o son refractarios a la deprivación androgénica por análogos LH-RH, con una baja morbilidad, buena aceptación de los pacientes y de muy bajo costo.

Introduction: The object of this clinical trial is to study the effectiveness of the administration of Diethylstilbestrol (DES) oral in patients with advanced cancer of prostate and that has presented resistance to the treatment with analogs LH-RH and in addition to evaluate the collateral effects attributable to its use. Material and methods: Between November of 2010 and May of 2012 15 patients with advanced prostate cancer, refractory to handling with analogs LH-RH entered themselves in consecutive form the study. Age average of the patients 69, 4 years, Rank 57-80. The type of made treatment was registered, detailing previous the hormonal handling which they were put under. Resistance to the analogs, the detection of 2 consecutive rises of the PSA was considered during the administration of this one. 1 mg. was indicated to the patients of DES to the day. The evaluation of the treatment was made every 30 days with determination of PSA and registry of collateral effects. Results: Of the 15 patients, 8 (53,3 percent) diminished their initial PSA, 5 of them (40 percent)did in more of a 50 percent of their value and 2 (13,3 percent) diminished the PSA but in less of a 50 percent. 7 patients (46,6 percent) did not diminished their value of PSA and were considered like failures. As collateral effects of the treatment we had 13 patients (86,6 percent) who presented hypersensitivity of the nipples, but single 2 required symptomatic treatment, 4 patients (26,6 percent) developed ginecomastia weighs moderate and we did not have any cardiovascular complication in the 15 studied patients. Conclusion: We considered according to our results that the oral DES is effective like treatment in the patients who have failed or ar refractory to the androgenic deprivation by analogs LH-RH, with a low morbidity, good acceptance of the patients and very low cost.

Synthesis of artificial diethylstilbestrol antigen for preparation of polyclonal antibodies / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To synthesize artificial diethylstilbestrol (DES) antigen and to prepare DES polyclonal antibody with high titer and sensitivity.</p><p><b>METHODS</b>The derivative of DES (DES-HS) was synthesized from diethylstilbestrol, ethyl bromoacetate,bovine serum albumin (BSA) and chicken ovalbumin (OVA) with the nucleophilic substitution reaction; the compound was identified by electrospray ionization mass spectrometry(ESI-MS). The DES-HS and the carrier proteins (BSA, OVA) were cross-linked to prepare the artificial antigen; the UV absorption spectrophotometry and high performance liquid chromatography (HPLC) were used to identify the prepared artificial antigen. The rabbits were immunized with the DES artificial antigen to prepare the DES polyclonal antibodies.</p><p><b>RESULTS</b>The DES-HS was synthesized. The DES artificial antigen was prepared successfully with a coupling rate of 22:1. The DES polyclonal antibodies with a titer of 1:25 600 and IC50 of 10.81 ng/ml were prepared with DES artificial antigen.</p><p><b>CONCLUSION</b>A set of methods to synthesize DES artificial antigen and to prepare the DES polyclonal antibodies has been developed successfully.</p>

Prevention of osteopenia and dyslipidemia in rats after ovariectomy with combined aspirin and low-dose diethylstilbestrol / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To study whether effect of aspirin plus low-dose diethylstilbestrol is more effective and safer than high diethylstilbestrol dose alone on prevention of ovariectomy-induced osteopenia and dyslipidemia.</p><p><b>METHODS</b>Thirty-eight 4-month-old female SD rats were divided into baseline (BAS) group (n=6), sham operation group (n=8) and ovariectomy (OVX) group (n=24). The OVX group was further divided into vehicle treatment group (n=8), diethylstilbestrol (30 μg/kg•d) treatment group (OVX+D30 group, n=8), and aspirin (9 mg/kg•d) plus diethylstilbestrol (10 μg/kg•d) treatment group (OVX+A-D10 group, n=8). Their left tibiae were collected for the bone histomorphometric analysis in undecalcified sections. Left femurs were collected for the bone mineral density measurement.</p><p><b>RESULTS</b>The body weight and serum cholesterol were increased, while uterine weight and cancellous bone mass were decreased in OVX rats compared with the SHAM group. Cancellous bone mass was significantly increased, while body weight and bone resorption parameters were decreased in both A-D10 and D30 treatment group compared with OVX group. The rats treated with A-D10 showed significantly increased in bone formation parameters and decreased in serum triglyceride compared with the D30-treated rats.</p><p><b>CONCLUSION</b>Aspirin plus low-dose diethylstilbestrol can effectively prevent osteopenia by reducing bone resorption, and is thus a better treatment modality for preventing dyslipidemia than high-dose diethylstilbestrol alone.</p>

Diethylstilbestrol affects LGR8 expression in mouse gubernaculum testis / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the impact of prenatal exposure to diethylstilbestrol (DES) on the specific receptor LGR8 of insulin-like factor 3 (INSL3) in the mouse gubernaculum testis, and that of exoestrogens on descensus testis in mice.</p><p><b>METHODS</b>A total of 120 pregnant KM mice aged 8 to 10 weeks were assigned to a normal, a blank control and 4 DES groups of equal number, the blank controls injected subcutaneously with dimethyl sulfoxide plus normal saline, and the DES groups with DES at 0.1, 1, 10 and 100 microg/kg body weight, respectively, from embryonic day 9 (ED9) through ED17. Immunohistochemistry and RT-PCR were used to detect the expressions of LGR8 protein and mRNA in the gubernaculum testis of the ED18 fetuses and PND20 (postnatal day 20) offspring of the mice.</p><p><b>RESULTS</b>Histological analysis showed that the gubernaculum testis of the ED18 fetuses were well developed in both the normal and control groups, with an inner mesenchymal core and muscular outer layer. In contrast, the gubernaculum testis were poorly developed in the experimental groups, morphologically abnormal and without visible dividing line between the mesenchymal tissue and the muscular outer layer. No obvious differences were found in the gubernaculum testis development of the neonates between the normal and experimental groups. Positive immunostaining was seen in the mesenchymal core and muscular outer layer, but mainly in the latter. The expression of LGR8 was weaker in the experimental groups than in the normal group (P < 0.05), but that of LGR8 mRNA was increased in the high-dose (10 and 100 microg/kg) DES groups (P < 0.05). No obvious mutations were observed in the PCR products in any of the experimental groups.</p><p><b>CONCLUSION</b>Prenatal exposure to diethylstilbestrol affected the expression of LGR8 mRNA in the mouse gubernaculum testis, which suggests that diethylstilbestrol may induce cryptorchidism by interfering with the INSL3-LGR8 signaling system and consequently the development of the gubernaculum testis.</p>

Expression of peroxiredoxin I regulated by gonadotropins in the rat ovary / 대한생식의학회지

OBJECTIVE: Peroxiredoxins (Prxs) play an important role in regulating cellular differentiation and proliferation in several types of mammalian cells. This report examined the expression of Prx isotype I in the rat ovary after hormone treatment. METHODS: Immature rats were injected with 10 IU of pregnant mare's serum gonadotropin (PMSG) to induce the growth of multiple preovulatory follicles and 10 IU of human chorionic gonadotropin (hCG) to induce ovulation. Immature rats were also treated with diethylstilbestrol (DES), an estrogen analogue, to induce the growth of multiple immature follicles. Northern blot analysis was performed to detect gene expression. Cell-type specific localization of Prx I mRNA were detected by in situ hybridization analysis. RESULTS: During follicle development, ovarian Prx I gene expression was detected in 3-day-old rats and had increased in 21-day-old rats. The levels of Prx I mRNA slightly declined one to two days following treatment with DES. A gradual increase in Prx I gene expression was observed in ovaries obtained from PMSG-treated immature rats. Furthermore, hCG treatment of PMSG-primed rats resulted in a gradual stimulation of Prx I mRNA levels by 24 hours (2.1-fold increase) following treatment, which remained high until 72 hours following treatment. In situ hybridization analysis revealed the expression of the Prx I gene in the granulosa cells of PMSG-primed ovaries and in the corpora lutea of ovaries stimulated with hCG for 72 hours. CONCLUSION: These results demonstrate the gonadotropin and granulosa cell-specific stimulation of Prx I gene expression, suggesting its role as a local regulator of follicle development.

Risk Factors for Breast Cancer, Including Occupational Exposures

The knowledge on the etiology of breast cancer has advanced substantially in recent years, and several etiological factors are now firmly established. However, very few new discoveries have been made in relation to occupational risk factors. The International Agency for Research on Cancer has evaluated over 900 different exposures or agents to-date to determine whether they are carcinogenic to humans. These evaluations are published as a series of Monographs (www.iarc.fr). For breast cancer the following substances have been classified as "carcinogenic to humans" (Group 1): alcoholic beverages, exposure to diethylstilbestrol, estrogen-progestogen contraceptives, estrogen-progestogen hormone replacement therapy and exposure to X-radiation and gamma-radiation (in special populations such as atomic bomb survivors, medical patients, and in-utero exposure). Ethylene oxide is also classified as a Group 1 carcinogen, although the evidence for carcinogenicity in epidemiologic studies, and specifically for the human breast, is limited. The classification "probably carcinogenic to humans" (Group 2A) includes estrogen hormone replacement therapy, tobacco smoking, and shift work involving circadian disruption, including work as a flight attendant. If the association between shift work and breast cancer, the most common female cancer, is confirmed, shift work could become the leading cause of occupational cancer in women.

Determination of longistylin A and longistylin C in Cajanus cajan / 中国中药杂志

<p><b>OBJECTIVE</b>To establish quality control criteria for medicinal herb Cajanus cajan based on the determination of longistylin A and longistylin C, two bioactive and specific stilbenes of the plant.</p><p><b>METHOD</b>Longistylin A and longistylin C were obtained from the leaves of C. cajan by silica gel column chromatography and identified as marker compounds of this plant by spectroscopic analysis. A RP-HPLC method was established to determine the two compounds.</p><p><b>RESULT</b>Longistylin A and longistylin C were well separated on a Thermo BDS Hypersil C18 column (4.6 mm x 250 mm, 5 microm) with a mobile phase methanol-water (8:2), and showed good linearity in the range of 0.00288 - 0.0576 microg and 0.0112 - 0.224 microg, respectively. The average recoveries were 98.9% and 97.2% with RSD of 2.4% and 2.2% for these two compounds, respectively.</p><p><b>CONCLUSION</b>The established analysis method is simple and accurate, whicn can be used for quality control of C. cajan.</p>

Expresion de la proteína P504S en las células prostáticas en la circulación sanguínea (CPCS) en pacientes con cáncer prostático y el efecto de la supresión adrogénica: estudio inmunocitoquímico / Circulating prostate cells(CPCS)in patient with prostate cancer express P504S and androgen blockage effect: an immunocytochemical based study

Determinar la presencia de CPCS en pacientes con cáncer prostático, la expresión de p504 S yel efecto de la supresión androgénica. Pacientes, materiales y método: en muestras de sangre venosa de 92 pacientes portadores de cáncer a la prostáta se separaron las células mononucleares por centrifugación diferencial. Las cpcs fueron identificadas utilizando anticuerpos monoclonales contra APE y P504S. Muestras de sangre de 10 mujeres fueron usadas como controles. Resultados: En ninguna de las muestras utilizadas como control y en el 68 por ciento de los hombres estudiados se detectaron CPCS. Todas las células detectadas fueron positivas para la expresión de P504S. Los pacientes con supresión androgénica, DES o después de una orquidectomía, tuvieron un nivel de P504S promedio menor que aquellos sin terapia sistémica p menor que 0,03. Conclusiones: la detección de CPCS P504S positivas en biopsias de prostáta es utilizada para el diagnóstico de cáncer, las celulas benignas no expresan este antígeno. Este estudio pionero demuestra que la expresión de P504S en CPCS es menor eb hombres con tratamiento hormonal sistémico.

Objective To determine the effect of androgen blockage on the expression of P504S en circulating prostate cells (CPCs) in men with prostate cancer. Patients, material and method: mononuclear cells were separated from venous blood using differential centrifugation and identification fied using monoclonal antibodies against PSA and P504S. 10 women were used as controls and 92 men with prostate cancer formesd the study group. Results: 64,8 percent of men were positive for CPCs, all the CPCs detected expressed the antigen P504S. No controls were positive. Conclusions. The detection of P504S postive cells in prostate biopsies is used to determine whether they are malignant or not, benign cells P504S negative. This is pioner study to show that CPCs are P504S positive, with the implication that they are malignant cells.

A case of clear cell carcinoma of uterine cervix not related to intrauterine dietylstilbesterol exposure / 대한산부인과학회지

Clear cell carcinoma of the uterine cervix is rare cancer that accounts for 4 to 9% of the adenocarcinoma of uterine cervix. Although intrauterine exposure to diethylstilbestrol (DES) during early pregnancy is one of the established risk factors, DES exposure may not be confirmed in all patients. We experienced a case of clear cell carcinoma in the uterine cervix of 67-year-old woman who was not exposed to DES. She was initially diagnosed as endometrial clear cell carcinoma because of the normal colposcopic finding and histologically proven clear cell carcinoma from endometrial aspiration biopsy and endocervical curettage. We performed a total laparoscopic hysterectomy with bilateral salpingo-oophorectomy and lymphadnectomy including both pelvic and para-aortic regions. On the final pathologic diagnosis of clear cell carcinoma confined to endocervix, the patient was received adjuvant concurrent chemoradiation with weekly cisplatin. We present the case with a brief review of related literature.

Effect of Benfotiamine on the Expression of Phosphodiesterase Type 5 and Its Isoforms in Type 2 Diabetic Rat Kidneys / 대한남성과학회지

PURPOSE: The phosphodiesterases (PDEs) are critical components in the cyclic AMP/protein kinase A and the cyclic GMP/phosphokinase G signaling pathways. The cAMP and cGMP pathways are regulated by activation and dissolution of PDEs. Benfotiamine, a lipophilic derivation of thiamine is known an activator of transketolase, is reported to prevent diabetic nephropathy by decreasing proteinuria and reducing oxidative stress. We did this study to investigate the effect of benfotiamine in type 2 diabetic rat kidneys. MATERIALS AND METHODS: We prepared 10 male Long-Evans Tokushima Fatty (LETO: control) and 20 male Otsuka Long-Evans Tokushima Fatty (OLETF) rats, which developed non-insulin-dependent diabetes mellitus (NIDDM) naturally. An oral glucose tolerance test confirmed diabetic development in the OLETF rats at 26 weeks. We classified 10 of the OLETF rats into Group I, the no treatment group and the other 10 into Group II, the treatment group. Group II received 100 mg/kg benfotiamine after developing DM. At 44 weeks, we checked kidney weight, serum glucose, free testosterone, insulin, total cholesterol, and triglyceride before sacrifice. We designed the primers for rat PDE5, PDE5A1, and PDE5A2 genes were carried out semiquantitive multiplex RT-PCR. Immunohistochemical staining was performed for monoclonal mouse anti-cGB-PDE5 and mouse monoclonal anti-smooth muscle alpha-actin. RESULTS: For the Control Group, Group I, and Group II, kidney weight was 2.13+/-0.23, 2.08+/-0.22, and 1.94+/-0.44 g; serum glucose was 279.50+/-56.79, 338.00+/-55.00, and 314.71+/-139.1 mg/dl; free testosterone was 1.46+/-1.08, 1.05+/- 0.42, and 0.72+/-0.56 pg/dl; insulin was 1.03+/-0.43, 1.09+/-0.83, and 1.15+/-1.08 ng/ml; total cholesterol was 86.83+/-4.79, 132.00+/-7.69, and 118.14+/-30.93 mg/dl; and triglyceride was 78.83+/-16.47, 177.83+/-75.62, and 194.57+/-92.57 mg/dl, respectively. All three groups expressed PDE5, PDE5A1, PDE5A2 mRNA, but Group I PDE5 mRNA expression was lower than that of Group C, II. However, the expression of PDE5A1 and PDE5A2 mRNA was not significantly different among the three groups. CONCLUSIONS: Serum cholesterol, triglyceride, and glucose were significantly higher in OLETF than in LETO rats. The PDEs were lower in diabetic rat (OLETF) kidneys and PDEs may play a significant role in the development of diabetic renal complications. Benfotiamine is suggested to increase expression of PDE5 mRNA in the type 2 diabetes rat kidney, but the difference in expression levels between PDE5A1 and PDE5A2 was not significant. These findings suggest that benfotiamine may play a specific role in diabetic changes of the rat kidney via a PDE5-related pathway, but it is not clear whether subtype PDE5A1 and PDE5A2 genes play a specific role.

Animal model for age- and sex-related genotoxicity of diethylstilbestrol

Environmental xenoestrogens pose a significant health risk for all living organisms. There is growing evidence concerning the different susceptibility to xenoestrogens of developing and adult organisms, but little is known about their genotoxicity in pre-pubertal mammals. In the present study, we developed an animal model to test the sex- and age-specific genotoxicity of the synthetic estrogen diethylstilbestrol (DES) on the reticulocytes of 3-week-old pre-pubertal and 12-week-old adult BALB/CJ mice using the in vivo micronucleus (MN) assay. DES was administered intraperitoneally at doses of 0.05, 0.5, and 5 µg/kg for 3 days and animals were sampled 48, 72 and 96 h, and 2 weeks after exposure. Five animals were analyzed for each dose, sex, and age group. After the DES dose of 0.05 µg/kg, pre-pubertal mice showed a significant increase in MN frequency (P < 0.001), while adults continued to show reference values (5.3 vs 1.0 MN/1000 reticulocytes). At doses of 0.5 and 5 µg/kg, MN frequency significantly increased in both age groups. In pre-pubertal male animals, MN frequency remained above reference values for 2 weeks after exposure. Our animal model for pre-pubertal genotoxicity assessment using the in vivo MN assay proved to be sensitive enough to distinguish age and sex differences in genome damage caused by DES. This synthetic estrogen was found to be more genotoxic in pre-pubertal mice, males in particular. Our results are relevant for future investigations and the preparation of legislation for drugs and environmentally emitted agents, which should incorporate specific age and gender susceptibility.

Fertility-sparing radical abdominal trachelectomy for cervical clear cell adenocarcinoma unrelated to diethylstilbestrol exposure : A case report / 대한산부인과학회지

Clear cell adenocarcinoma (CCAC) is a rare cancer that comprises less than 9% of the cervical adenocarcinoma cases. We experienced a case of fertility-sparing radical abdominal trachelectomy for cervical clear cell adenocarcinoma (CCAC). Thus, reported it. A 27 year old female was diagnosed with clinical stage Ib cervical CCAC. She had no history of maternal exposure to diethylstilbestrol and had negative PAP cytology and HPV tests. She was treated with neoadjuvant chemotherapy followed by radical abdominal trachelectomy. After 2 cycles of postoperative adjuvant chemotherapy, the lesion disappeared completely in an imaging study, and potential fertility was preserved. Radical abdominal trachelectomy with chemotherapy may be a valuable approach for treating stage Ib cervical CCAC in women that wish to preserve potential fertility.