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This study aims to examine the expression of p53, p16, and murine double minute 2 (MDM2) protein in normal endometrium and endometriosis, in order to discuss the role of p53, p16, and MDM2 protein and apoptosis in the pathogenesis and development of endometriosis, and provide a theoretical basis for clinical diagnosis and treatment. The immunohistochemical streptavidin-biotin peroxidase method was used to detect the expression of p53, p16, and MDM2 in tissue samples obtained from 30 women with pathologically confirmed ovarian endometriosis and 29 women with pathologically confirmed normal endometrium. The relationship between p53, p16, and MDM2 expression and apoptosis was analyzed. In normal endometrium, the positive rate of p53 in the secretory phase was higher than that in the proliferative phase (P .05). As important apoptosis regulatory genes, p53, p16, and MDM2 might be involved in the pathogenesis and development of endometriosis.

Purpose: To investigate the effect of thiazolidinedione amide (TZDA) treatment on high-sensitivity Creactive protein (hsCRP) levels and endothelial dysfunction in patients with polycystic ovary syndrome (PCOS). Methods: Twenty five women (mean age 24.7 ± 3.9 years; mean body mass index (BMI), 23.2 ± 4.0 kg/m 2 ) with PCOS were treated with 15 μM TZDA daily for 12 months. Serum levels of testosterone, leutenizing hormone (LH), follicle stimulating hormone (FSH), sex hormone-binding globulin (SHBG), insulin and hsCRP were measured. BMI, hirsutism scores and insulin sensitivity indices were also calculated prior to and after TZDA treatment. Brachial artery responses to stimuli was used to measure arterial endothelium and smooth muscle function prior to and after the treatment. Results: TZDA treatment caused a significant (p < 0.05) decrease in serum testosterone from 93.1 ± 40.3 to 54.8 ± 19.5 ng/dl and fasting insulin concentration from 11.9 ± 6.8 to 9.23 ± 5.13 U/mL. Insulin resistance index significantly (p < 0.05) improved and hirsutism score decreased significantly from 11.6 ± 2.0 to 6.8 ± 2.0. BMI, waist circumference, serum total cholesterol, low-density lipoprotein (LDL)- cholesterol, FSH and LH levels remained almost unchanged. Twenty-four of the women reverted to regular menstrual cycles. SHBG levels showed a significant (p < 0.05) increase from 24.8 ± 9.5 to 49.1 ± 13.5 nmol/L after TZDA treatment. Serum hsCRP levels decreased (p < 0.05) from 0.25 ± 0.1 to 0.09 ± 0.02 mg/dL while endothelium-dependent vascular responses significantly improved (p < 0.05) following TZDA treatment (9.9 ± 3.9 vs 16.4 ± 5.1%). Conclusion: TZDA treatment improves insulin sensitivity, decreases androgen production and improves endothelial dysfunction in young women with PCOS. Keywords: Thiazolidinedione amide, Insulin sensitivity, Endothelial dysfunction, Polycystic ovary syndrome

Ovarian cancers are often at an advanced stage at diagnosis because early detection is difficult. The poor prognosis of ovarian cancers highlights the crucial need to develop better therapeutic agents and strategies. The objective of this study was to investigate the inhibitory effects of a new modified antisense oligonucleotides targeting vascular endothelial growth factor A (VEGF-A) in SKOV3 ovarian cancer cells.Antisense oligonucleotides targeting VEGF-A was designed, synthesized and transfected into SKOV3 ovarian cancer cells. Western blotting and real-time RT-PCR were used to analyze the inhibitory effects of antisense oligonucleotides on VEGF-A protein and mRNA expression. Transwell matrix assay was used to detect cell migration inhibition.The antisense oligonucleotides targeting VEGF-A significantly decreased VEGF-A protein and mRNA expression and inhibited cell migration in SKOV3 ovarian cancer cells.This new modified antisense oligonucleotides targeting VEGF-A can decrease VEGF-A expression and inhibit cell migration in SKOV3 ovarian cancer cells. This new oligonucleotides may be a promising therapeutic agent for ovarian cancers.

Using Longissimus Dorsi muscle as material and Lambda ZAP II as Vector, Xiang Pig Longissimus Dorsi muscle cDNA library has been constructed in our study. The results showed that the titration of the library was 3.4 x 10(7) pfu/ml, the recombinant percentage was 94%, and the fragment length of inserted average cDNA were 1.5 kb. The study pointed out that the more than 30 T insertion is the major factor for low percentage if sequencing the 3'-end.

For the first time, a 16 kb fragment of the porcine Ob gene, namely intron1, exon1, 5' region of Ob gene, was restrictively analyzed and sequenced by the primers designed in the portion of the swine Ob sequence. The first small 38 bp untranslated exon1 is located 11.1 kb upstream of the initiator ATG codon, and two novel microsatellites SW200 and SW160 are found in intron1. Promoter analysis of several putative binding sites revealed that this 300 bp promoter located at -1 to -300, including C/EBP and two Sp1, may be as effective as the longer promoter in directing leptin transcription. To examine microsatellites association with important economic traits, we statistically analyzed genotypes and alleles of the two microsatellites. Statistical analysis carried out by SAS 8.2 revealed significant positive correlation between the two microsatellites genotypes and the litter size in first parity of Erhualian.

To evaluate the effects of antiprogestins mifepristone and lilopristone on proliferation and expressions of nuclear factor-kappa B (NF-kappaB) of ectopic stromal cells in vitro.The ectopic stromal cells of ovary were cultured in vitro. Methyl thiazolyl tetrazolium method was used to evaluate proliferative activity of ectopic stromal cells. Cells were divided into five groups according to drug concentration: control group, group I and group II of mifepristone and of lilopristone. The expressions of NF-kappaB P65 and NF-kappaB P65 mRNA of ectopic stromal cells were determined by immunocytochemistry and in situ hybridization of cell slice.Antiprogestins mifepristone and lilopristone were able to significantly suppress the proliferation of ectopic stromal cells in a time- and dose-dependent manner in vitro. The expressions of NF-kappaB P65 and NF-kappaB P65 mRNA of ectopic stromal cells in the control group were higher than that of group I and group II. Their expressions in mifepristone groups were higher than that of lilopristone groups.Antiprogestin mifepristone and lilopristone could significantly suppress the proliferation of ectopic stromal cells in a time- and dose-dependent manner in vitro. The action mechanisms may be associated with the suppression of expression of NF-kappaB P65 mRNA and NF-kappaB P65.

The variation of chromosomes of Gansu Black pig was caused by infusion of Duroc blood. The chromosome length ratios of No.1,6,7,13 chromosomes in the line I of Gansu Black pig were higher than those in the line II. It showed that the homogeneity in line I was higher than in line II. The number of Ag-staining nucleolus organizers (Ag-NOR) was between 1-4 in the line I and II, the average No. of Ag-NOR was different obviously between the line I and the line II. This indicated that the activity of translation in rRNA in the line I was different in the line II.

In animal breeding, microsatellite marker plays an important role in constructing genetic maps, QTL mapping and function analysis of structural genes. Myostatin, also known as GDF8, is a negative regulator of skeletal muscle mass and, in swine, it is evidenced to be related to birth weight and average daily gain from 60 kg to 100 kg of body weight. In present study, by subcloning and sequencing,we identified a novel microsatellite marker which is useful for fine QTL mapping for meat traits. A BAC clone containing porcine MSTN was extracted and digested with EcoR I to recover the fragment of4 kb for subcloning in pGEM-3zf (+). Sequencing and alignment results showed that this subcloned fragment was not from porcine MSTN, but included a tandem repeat of (TG) 13, which is a novel microsatellite marker (GenBank accession number: AF454400) flanking MSTN. To exclude its vector origin we designed specific primers flanking this marker and successfully amplified this fragment from porcine genome. Through a pedigree analysis of a double-muscled Yorshire strain, we found that it is inherited in a co-dominant manner. We also checked the gene frequencies of this locus in 381 unrelated individuals of 7 pig breeds, namely Laiwu,Landrace, Yorkshire,Duroc, Peterian, Min and Erhualian. Only two alleles were detected, the repeating number of which are 13 (allele A) and 19 (allele B) respectively, which indicated that it is a low poly morphic microsatellite marker. In addition, the frequencies of the two alleles are different between the two types of pig breeds, while allele A is dominant in Chinese local breeds, allele B is dominant in imported breeds. Alignment with AY208121 indicate that this locus is located 42 kb downstream of porcine MSTN. We speculate that this microsatellite DNA is an important marker both in fine QTL mapping for meat traits and in the expression study of porcine MSTN.

We constructed the first microsatellite-enriched library of yak according to the strong affinity between biotin and streptavidin. The method included ligation of 300 approximately 1 000 bp enzyme-digested fragments and adaptors, affinity capture of microsatellite repeat using biotinylated oligoprobe ((CA)12, (CCG) 8, (CAG)8, (TTTC) 8) attached to streptavidin-coated magnetic beads, PCR amplification using the 21-mer adaptor oligonucleotide as primer to obtain double-stranded targeted fragments, religated into pMD18-T vector and transformed to DH5alpha. The results of sequencing showed that 37 of 48 readable sequences contained microsatellites indicating a high degree of microsatellite enrichment. The new polymorphic microsatellite markers we have identified and characterized will contribute to the yak genetic linkage mapping, molecular evolution and phylogenetic studies, marker assistant selection and QTLs location of yak main economic traits.

Observations on structure and distribution of vimentin as determined by indirect immunofluorescence were showed that the arrangement of vimentin intermediate filments (IFs) changed from an extended fibrillar state to a complex cage formation tightly associated with the forming lipid globules during adipose conversion of rat preadipocytes. The effects of differentiation of rat preadipocytes into adipocytes on vimentin mRNA levels as determined by in situ hybridization (ISH) using oligonucleotides probes to vimentin mRNA labeled by digoxigenin (DIG), and on vimentim protein expression as determined by Western blotting were researched. The results suggested that the expressions of vimentin with molecular weight of about 57 kD at the levels of both mRNA and protein was existed throughout the predipocyte differentiation and downregulation of vimentin is required for adipose conversion. The specificity of the association of lipid globules with vimentin IFs during adipose conversion was taken as an example of a possible IF function in relation to a preadipocytes differentiation process, especially discussed as a functional relationship in supporting adiposgenesis.

To investigate the effect of mifepristone on the activity of proliferation and the apoptosis, the expression of estrogen receptor (ER), progesterone receptor (PR) protein and morphology changes of human ovarian carcinoma cell line 3AO and SKOV3 in vitro.The proliferative activity of 3AO and SKOV3, which were cultured in vitro, was measured by tetrazolium-based colorimetric assay (MTT assay). Flow cytormetry (FCM) was used to measure the expressive rate of ER, PR, p53 protein, bcl-2 protein, cell apoptotic rate and cell proliferative cycle of 3AO cells, which were cultured with different concentration and duration of mifepristone. The morphologic and ultrastructure changes of apoptotic 3AO cells was observed by the light and electron microscopy.Mifepristone inhibited significantly the proliferation of 3AO cells in dose-time dependent manner in vitro. The inhibitory rate of 3AO cells growth, which were cultured with different concentration of mifepristone (5, 10, 20, 40, 80 micro mol/L) and duration (24, 48, 72 h) was from 1.7% to 75.0% (P0.01), but the proliferation activity of SKOV3 cells in vitro had not significant changes (P0.05). 3AO cells apoptosis activity appeared the positive correlation with the dose of mifepristone and cultured duration (P0.01). Mifepristone blocked 3AO cells proliferative cycle at the G(0)-G(1) stage, decreased the cell radio of S stage. Mifepristone induced the apoptosis of 3AO cells in vitro. The light and electron microscopy demonstrated that 3AO cells cultured with mifepristone appeared typical morphological characteristics of apoptosis including the compaction and margination of the chromosomes, nuclear fragments and formation of apoptotic bodies. Mifepristone up-regulated significantly the expression of p53 protein, but down-regulated the expression of bcl-2 protein (P0.01). The expressive rates of p53 and bcl-2 proteins were (54.8 +/- 4.0)% and (10.1 +/- 1.2)%, respectively, when 3AO cells was cultured with mifepristone of 10 micro mol/L for 24 h. Compared with the expression rates of control group (27.1 +/- 3.3)% and (17.6 +/- 1.0)%, the difference was significant (P0.01). The expressive rate of PR protein of 3AO cells cultured with mifepristone of 10 micro mol/L for 48 h was (12.7 +/- 1.4)%, which was decreased compared with the expressive rate of control group (44.9 +/- 1.4)% (P0.01).Mifepristone inhibited significantly the proliferation and induced the cell apoptosis of cell line 3AO in dose-time dependent manner in vitro. The anti-tumor effect was related to down-regulation the expression of PR protein and bcl-2 protein, and to up-regulation the expression of p53 protein of 3AO cells.

In this experiment, F2 chicken derived from Broilers crossing with Silky was used to study the effect of insulin-like growth factor-II gene on growth and carcass traits. The partial gene was amplified by two pairs of primers, and single nucleotide polymorphism (SNPs) was detected by the technique of restriction fragment length polymorphism (RFLP), and then confirmed by DNA sequencing. The mutation was found in the exon-2 of the gene, and can be clarified by cutting of restriction enzyme Aci-I. The result of least square analysis showed the gene was significantly related with growth and carcass traits. It implied that the insulin-like growth factor-II gene could be a genetic locus or linked to a major gene affecting greatly the growth and carcass traits in chicken.