OBJECTIVETo investigate the effects of mono(2-ethylhexyl) phthalate(MEHP), the primary metabolite of di(2-ethylhexyl) phthalate (DEHP), on testosterone biosynthesis in Leydig cells cultured from the Sprague Dawley rat testis.

METHODSBased on the primary Leydig cell culture model, MEHP exposure groups involved control (0 micromol/L), 62.5, 125, 250, 500 and 1000 micromol/L. We observed mitochondria activity with the MTT method, measured the testosterone level with RIA and determined steroidogenesis acute regulatory protein (StAR) mRNA expression with RT-PCR.

RESULTSAfter Leydig cells were exposed to MEHP for 24 hours, the activity of mitochondria enhanced evidently at 250 micromol/L and then declined markedly at 1000 micromol/L compared with the control group (P < 0.01). The testosterone level showed an increasing tendency in both basal and hCG-stimulated states with statistical significance at 250 and 500 micromol/L compared with the control group (P < 0.01). However, the expression of StAR mRNA appeared unchanged at 62.5, 125 or 250 micromol/L, but exhibited a decreasing tendency at 500 and 1000 micromol/L (P < 0.01).

CONCLUSIONME- HP directly affected the activity of mitochondria and testosterone biosynthesis of the Leydig cells in vitro. StAR, the regulator of cholesterol transport into mitochondria, might not be responsible for the increase of testosterone biosynthesis induced by MEHP.

Animals ; Cells, Cultured ; Diethylhexyl Phthalate ; analogs & derivatives ; pharmacology ; Dose-Response Relationship, Drug ; Leydig Cells ; drug effects ; metabolism ; Male ; Phosphoproteins ; biosynthesis ; genetics ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; Testosterone ; biosynthesis

OBJECTIVEThe present study was performed to examine functional and structural impairment of rat sertoli cells following dibutyl phthalate (DBP) exposure.

METHODSThe 6-week-old healthy male Sprague Dawley rats were randomly divided into 4 groups with 16 animals in each group. DBP dissolved in peanut oil was administered by gavage at doses of 0, 250, 500 and 1 000 mg/kg. After 2-week DBP treatment, half of the rats were sacrificed. The rest were killed following 4-week DBP exposure. Follicle stimulating hormone (FSH) was analysed by radioimmunoassay. The relative expression levels of androgen binding protein (ABP) mRNA and inhibin (INH)alpha mRNA were determined by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR). The sertoli cell ultrastructures were observed by using transmission electron microscope (TEM).

RESULTSFSH levels were increased after 4-week DBP exposure with significance at doses of 250 and 1 000 mg/kg. Sperm head count and daily sperm product were decreased significantly in 500 and 1 000 mg/kg groups. The expression levels of ABP mRNA were 0.89 +/- 0.15, 0.85 +/- 0.23, 0.54 +/- 0.17, 0.52 +/- 0.16 and 0.88 +/- 0.16, 0.61 +/- 0.12, 0.48 +/- 0.15, 0.47 +/- 0.11 for 0, 250, 500 and 1 000 mg/kg after 2- and 4-week DBP treatments respectively with significance at doses of 500 and 1 000 mg/kg (P < 0.01), while the levels of INHalpha mRNA were 0.88 +/- 0.16, 0.61 +/- 0.12, 0.48 +/- 0.15, 0.47 +/- 0.11 and 0.75 +/- 0.19, 0.56 +/- 0.16, 0.53 +/- 0.08, 0.45 +/- 0.10 with significance at all exposure groups (P < 0.01 or P < 0.05). In sertoli cells of rats exposed to 1 000 mg/kg DBP, TEM photos showed more lysosomes in cytoplasm, proliferated and expanded endoplasmic reticulum and nuclei malformation.

CONCLUSIONSSertoli cell should be one of the major toxic targets. Impairment of spermatogenesis caused by DBP should be partly due to the suppression of ABP and INHalpha biosynthesis.

Androgen-Binding Protein ; genetics ; Animals ; Dibutyl Phthalate ; administration & dosage ; toxicity ; Dose-Response Relationship, Drug ; Follicle Stimulating Hormone ; metabolism ; Inhibins ; genetics ; Male ; RNA, Messenger ; genetics ; metabolism ; Radioimmunoassay ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; Sertoli Cells ; drug effects ; metabolism ; pathology ; Testis ; drug effects ; metabolism ; pathology

OBJECTIVETo detect the hypermethylation status of RASSF1A promoter in serum DNA of non-small cell lung cancer (NSCLC) patient and evaluate its correlation with clinicopathological parameters.

METHODSSerum DNA was extracted from the peripheral blood of 75 NSCLC patients and another 35 patients with benign pulmonary disease and 15 healthy donors. The methylation status of RASSF1A promoter was determined using methylation-specific PCR (MSP), and the correlation of methylation profiles with clinicopathological parameters was statistically analyzed.

RESULTSAberrant methylation of RASSF1A was detected in 23 of 75 (30.7%) cancer patients, but in none of patients with benign pulmonary disease or in healthy donors (P <0.001). RASSF1A hypermethylation status was found to be correlated with late stage and poor differentiation (P < 0.05), but not with gender, age or histopathology in NSCLC patients.

CONCLUSIONHypermethylated RASSF1A promoter is frequently found in the serum DNA of non-small cell lung cancer patient, and RASSF1A may become a promising novel biomarker for diagnosis and prognosis prediction in lung cancer.

Adult ; Aged ; Biomarkers, Tumor ; blood ; Carcinoma, Non-Small-Cell Lung ; blood ; genetics ; pathology ; Case-Control Studies ; DNA Methylation ; DNA, Neoplasm ; blood ; genetics ; Female ; Humans ; Lung Neoplasms ; blood ; genetics ; pathology ; Male ; Middle Aged ; Neoplasm Staging ; Polymerase Chain Reaction ; methods ; Promoter Regions, Genetic ; Tumor Suppressor Proteins ; blood ; genetics

OBJECTIVETo analyze the incidence and time trends of esophageal and gastric cancers in Linzhou city bassed on the data of Linxian Tumor Registry, and to provide valid reference data for research and effective estimation of cancer control in this area.

METHODSAll incidence records for the both cancers during 1988-2003 were drawn from Linzhou Tumor Registry and grouped by sex, age, year and then linked to corresponding population data. The incidence rates of those two topographic site cancers were calculated and the age-adjusted rates were calculated by direct standardization to the world population. A joinpoint model was used to get the annual percentage change (APC) of the age-adjusted rates, and to estimate the epidemiological trends of both cancers in population of Linzhou city.

RESULTSIn the year 2003 the age-adjusted incidence rates of esophageal and gastric cancers were 81.78 per 100 000 and 77.08 per 100 000, respectively, in the population of Linzhou city. The incidence rate of both cancers showed a decreasing trend from 1988 to 2003. The APC of the incidence rates of esophageal cancer was - 2.6% and that of gastric cancer was - 1.8%, and both indexes were statistically significant (P < 0.05).

CONCLUSIONThe incidence rates of esophageal and gastric cancers have presented a decreasing trends in the population of Linzhou city. This trend will continue along with the development of social economy, elevation of living standard and improvement in living habit and environment.

Cardia ; China ; epidemiology ; Esophageal Neoplasms ; epidemiology ; Female ; Humans ; Incidence ; Male ; Sex Factors ; Stomach Neoplasms ; epidemiology

OBJECTIVETo investigate the gene prevalence and spectrum of alpha- and beta-thalassemia in Fujian province.

METHODSA total of 11 234 of neonatal cord blood samples were collected for a prevalence study of alpha- and beta-thalassemia. All subjects included in this study were registered in 9 cities of Fujian province. A complete blood count and high performance liquid chromatography (HPLC) were performed in all samples, with microcytosis (MCV≤ 79 f1 and MCH≤ 27 pg) or HPLC positive cases further studied by DNA analysis. alpha- and beta-thalassemia were determined by using gap-PCR and reverse dot blot (RDB) assays. Unknown positive samples were analyzed directly with DNA sequencing.

RESULTSOf all 11 234 cord blood samples, 356 were identified as from alpha-thalassemia gene carriers, 7 deletion genotypes were identified including 236 (--SEA/ α α) cases, 67 (α 3.7/ α α) cases, 24 (alpha 4.2/alpha alpha) cases, 3 (alpha 3.7/ SEA) cases, 1 (alpha 4.2/ SEA) cases, 1 (alpha 3.7/ alpha 3.7) cases, 1 (alpha 3.7/ alpha 4.2) cases; 3 non-deletion genotypes were detected, including 7 (alpha alpha QS/ alpha alpha) cases, 3 (α α CS/α α) cases, 2 (α α WS/ α α) cases, the most common mutation was SEA/α α, which accounted for 66.29%, 148 individuals were found to have beta-hemoglobin gene mutations. 12 different mutations were identified, namely 65 IVS-2 654 (C>T) cases, 40 CD41-42(-TCTT, 12 CD17(A>T) cases, 10 -28(A>G) cases,7 CD27-28(+C) cases, 5 start codon ATG>AGG cases, 2 CD26(G>A) cases, 1 CD71-72(+A) cases, 1 IVS-1-1(G>T) cases, 1 CD43(G>T) cases, 2 -29(A>G) cases, 2 Codon 36 (-C) cases, the most common mutation was IVS-2 654(C>T) and CD41-42(-TCTT), which accounted for 70.95%. A novel beta-globin gene mutation CD36 (-C) allele was also detected. The carrier rate of thalassemia in Fujian population is 4.41%. In addition, 9 beta-thalassemia carriers were found with alpha-thalassemia mutation.

CONCLUSIONThe research has revealed the type of gene mutations in alpha- and beta-talassemia in Fujian province. The beta-thalassemia mutations in Fujian province are complex, which were also obviously heterogeneous. This will significant value for screening the incidence, provide the valuable information for genetic counseling and prenatal diagnosis.

Adolescent ; Adult ; China ; epidemiology ; Female ; Genotype ; Humans ; Male ; Middle Aged ; Prevalence ; Young Adult ; alpha-Thalassemia ; epidemiology ; genetics ; beta-Globins ; genetics ; beta-Thalassemia ; epidemiology ; genetics

OBJECTIVETo evaluate the chromium (Cr) levels in blood and urine among general population in China between 2009 and 2010, and thereby to analyze its prevalent features.

METHODSFrom year 2009 to 2010, a total of 11 983 subjects of general population aged between 6 and 60 year-old were recruited from 24 districts in 8 provinces in eastern, central and western China mainland, by cluster random sampling method. The information about their living environment and health status were collected by questionnaire, and 11 983 blood samples and 11 853 urine samples were also collected. Inductively coupled plasma mass spectrometry (ICP-MS) was applied to test the Cr level both in blood and urine; and the Cr distribution in blood and urine among groups of population in different ages, genders and districts, were then analyzed.

RESULTSAmong general population in China, the geometric mean (GM) of Cr concentration in blood was 1.19 µg/L, with median at 1.74 µg /L and 95% percentile at 5.59 µg/L. The Cr concentration in blood among males and females were separately 1.18 µg/L and 1.20 µg/L(P > 0.05); while its GM in the groups of population aged 6 - 12, 12 - 16, 16 - 20, 20 - 30, 30 - 45 and 45 - 60 years old were 1.00, 1.22, 1.01, 1.40, 1.27 and 1.30 µg/L (P < 0.01), respectively; and the figures in populations from eastern, central and western China were 1.00, 1.70 and 1.98 µg/L (P < 0.01), respectively. Among general population, the GM of Cr concentration in urine was 0.53 µg/L, with median was lower than 0.42 µg/L and 95% percentile at 3.53 µg/L. The Cr concentration in urine among males and females were separately 0.52 µg/L and 0.53 µg/L (P > 0.05);while its GM in the groups of population aged 6 - 12, 12 - 16, 16 - 20, 20 - 30, 30 - 45 and 45 - 60 years old were 0.56, 0.60, 0.52, 0.50, 0.52 and 0.46 µg/L (P < 0.01), respectively;and the figures in populations from eastern, central and western China were 0.58, < 0.42 and 0.60 µg/L (P < 0.01), respectively.

CONCLUSIONThe study reported the Cr levels in blood and urine among general population in China, and thereby provided basic data evidence for the following Cr biological monitoring studies in near future.

Adolescent ; Adult ; Child ; China ; Chromium ; blood ; urine ; Female ; Humans ; Male ; Middle Aged ; Population Surveillance ; Young Adult

BACKGROUNDAt present, a diagnostic tool with high specificity for impaired endometrial receptivity, which may lead to implantation failure, remains to be developed. We aimed to assess the different endometrial microRNA (miRNA) signatures for impaired endometrial receptivity by microarray analysis.

METHODSA total of 12 repeated implantation failure (RIF) patients and 10 infertile patients, who conceived and delivered after one embryo transfer attempt, were recruited as RIF and control groups, respectively. Endometrial specimens from the window of implantation (WOI) were collected from these two groups. MiRNA microarray was conducted on seven and five samples from the RIF and control groups, respectively. Comparative, functional, and network analyses were performed for the microarray results. Quantitative real-time polymerase chain reaction (PCR) was performed on other samples to validate the expression of specific miRNAs.

RESULTSCompared with those in the control group, the expression levels of 105 miRNAs in the RIF group were found to be significantly up- or down-regulated (at least 2-fold) by microarray analysis. The most relevant miRNA functional sets of these dysregulated miRNAs were miR-30 family, human embryonic stem cell regulation, epithelial-mesenchymal transition, and miRNA tumor suppressors by tool for annotations of microRNA analysis. Network regulatory analysis found 176 miRNA-mRNA interactions, and the top 3 core miRNAs were has-miR-4668-5p, has-miR-429, and has-miR-5088. Expression levels of the 18 selected miRNAs in new samples by real-time PCR were found to be regulated with the same trend, as the result of microarray analysis.

CONCLUSIONSThere is a significant different expression of certain miRNAs in the WOI endometrium for RIF patients. These miRNAs may contribute to impaired endometrial receptivity.

Adult ; Embryo Implantation ; genetics ; physiology ; Endometrium ; metabolism ; Female ; Humans ; Infertility, Female ; genetics ; MicroRNAs ; genetics ; Microarray Analysis ; Pregnancy ; Real-Time Polymerase Chain Reaction