Effect of Semen vaccariae and Taraxacu mogono on Cell Adhesion of Bovine Mammary Epithelial Cells

The aim of this study is to reveal the regulation mechanism of the effect of Semen vaccariae and Taraxacu mogono on the cell-cell adhersion molecule, E-cadherin and β-catenin on the proliferation role and secretion function of bovine mammary epithelial cells cultured in vitro. Firstly, the epithelial character of bovine mammary epithelial cells was authenticated using immunofluorescence, then the cell grow curve was observed and investigated after S. vaccariae and T. mogono treatment. On the effect of S. vaccariae and T. mogono, cell adhesion molecules E-cadherin, β-catenin and CycinD1 mRNA and protein were detected by qRT-PCR and Western blotting, respectively. The results showed that the cellular keratin 18 expressed positively and proliferfated vigorously after S. vaccariae and T. mogono treament. The mRNA and protein levels of E-cadherin and CycinD1 were remarkably higher （P〈0.05） in 36 h after S. vaccariae and T. mogono treatment. The cell proliferation at 36 h was increased significantly （P〈0.05）. In conclusion, S. vaccariae and T. mogono have a positive impact on the cell proliferation and an effect on the adhesion molecules E-cadherin, β-catenin and CycinD1 in the Wnt signaling pathway.

Bzw2 Promotes Proliferation and Lactation of Mammary Epithelial Cell in Dairy Goat

Mitosis of mammary epithelial cell is foundation of mammal lactation. We developed a strategy of combined application of generation of longer cDNA fragments from the serial analysis of gene expression （SAGE） tags for gene identification （GLGI） to screen and identify genes influencing lactating ability of mammary epithelial cell in dairy goat. GLGI as a new tag identification technique was brought about with SAGE. Bzw2 was found as a candidate gene related to lactation by screening Long-SAGE library of mammary gland in dairy goat. Bzw2 cDNA was synthesized by switching mechanism at 5＂-end of RNA transcript （SMART） technology. The mRNA level of Bzw2 was relatively higher in early lactation than in other development stages of mammary gland. The proliferation of mammary epithelial cell was inhibited by transfecting specific shRNA of Bzw2. The mRNA levels of Stat5, Csn2 and Prlr were also down-regulated, suggesting the lactating ability of mammary epithelial cell was attenuated after Bzw2 RNAi. The reduction of mammary epithelial cell growth and lactation by Bzw2 RNAi was rescued through over-expression of Bzw2. These results revealed that Bzw2 might play an important role in lactation though the molecular mechanism was still unclear.

The Effects of Insulin and Prolactin on an Epithelial Cell Line from Mammary Gland of Dairy Goat

Mammary epithelial cells with lactational function can be a valuable cellular model for research of the development and regulation of the mammary gland.This paper describes some aspects of function of an epithelial cell line from the mammary gland of the dairy goat.SDS-PAGE,triglyceride and lactose content of cultured cells were used to assess synthetic function of cells and the effects of exposure to insulin and prolactin.Results show that goat mammary epithelial cells can synthesize fat,proteins and lactose when they were cultured in DMEM-F12 medium with added EGF,IGF-1,ITS and FBS.There were no obvious changes after 48h treatment with additional insulin.Prolactin added to the basal medium significantly increased synthesis of proteins and lactose.A mammary gland epithelial cell line from goats which has lactational function has been established.This outcome provides a valuable and convenient model system.

Proteomic Identification of Differentially Expressed Proteins in Vaccaria segetalis-treated Dairy Cow Mammary Epithelial Cells

To investigate the potential effects of Vaccaria segetalis, we employed the proteomic approach on the dairy cow mammary gland epithelial cells （DCMECs）. A total of 35 differentially expressed nuclear proteins were visualized by 2-DE and silver nitrate staining. Of these, five proteins also displayed significant expression changes upon treatment of the water decoction from Vaccaria segetalis, and such alterations were further confirmed by RT-PCR. Together, at both the mRNA and protein levels, the water decoction from Vaccaria segetalis increased the expression of proteins ethylmalonic encephalopathy 1 （ETHE1）, vesicle amine transport protein 1 homolog （VAT1）, parkinson disease protein 7 homolog （Protein DJ-1）, proteasome subunit alpha type-2 （PSMA2） and SUMO-activating enzyme subunit 1 （SAE1）. This study would enable a better understanding of the molecular mechanisms underlying this water decoction effects at the protein level.

Galactopoietic Activity of Dibutyl Phthalate Isolated from Vaccaria segetalis

A galactopoietic compound, identified as dibutyl phthalate（DBP）, was isolated from Vaccaria segetalis. The activity of DBP on lactation ability of dairy cow mammary gland epithelial cells（DCMECs） cultured in vitro and dairy cow was evaluated. Results showed that DBP could promote cell viability, proliferation ability, lactose and β-casein secretion of DCMECs, which could also raise the milk yields of dairy cows significantly.

Screen of Bovine Mammary Gland Epithelial Cell Specifcity Promotor

Three lactoproteins （α-Sl-casein, β-lactoglobulin, and β-casein） promotors were cloned, sequenced and compared relative luciferase expression. The results showed that the promotor activity of bovine α-S1-casein gene was the best, and would be used to produce pharmaceutically and medically important proteins in the mammary gland of transgenic animals and also for the construction of an inducible eukaryotic expression vector.

Impact of let-7g on Proliferation and Lactation of Mouse Mammary Epithelial Cells

let-7g, a member of the let-7 family, regulates gene expression at the post-transcriptional level. The study explored a series of biological effects of mouse mammary epithelial cells that let-7g was produced. The differential expression of let-7g was detected by qRT-PCR in different developmental stages of the mouse mammary gland, let-7g expression and impact of let-7g on mouse mammary epithelial cells were analyzed by CASY-technology, qRT-PCR, Western blotting and HPLC inhibited let-7g expression of mouse mammary epithelial ceils through gene silencing. The results showed that qRT-PCR identified let-7g as being down-regulated in mouse mammary epithelial cells after it was inhibited. Mouse mammary epithelial cells with low expression of let-7g displayed higher expression of TGFβR I protein than those with high expression of let-7g, suggesting that low let-7g expression contributed to TGFβR I over-expression. Finally, the expression of let-7g was down-regulated, which significantly enhanced the proliferation of mouse mammary epithelial cells, and increased expression of β-Casein. The data indicated that let-7g could negatively regulate the expression of target Tgfbrl by complementary combination in mouse mammary epithelial cells, and then regulate the cell proliferation and expression of β-Casein by suppressing the TGFβR I expression.

Proteomic Analysis of Nuclear Phosphorylated Proteins in Dairy Cow Mammary Epithelial Cells Treated with Prolactin

Prolactin （PRL） is a versatile signaling molecule and regulates a variety of physiological processes, including mammary gland growth and differentiation and the synthesis of milk proteins. While PRL is known to be necessary for high levels of milk protein expression, the mechanism by which the synthesis of milk proteins is stimulated at the transcript level is less known. A major modification in the transcript level is protein phosphorylation. To gain additional insights into the molecular mechanisms at the transcript level underlying PRL action on the dairy cow mammary epithelial cells （DCMECs）, nuclear phosphoproteins whose expression distinguishes proliferating regulated by PRL in DCMECs were identified. A phosphoprotein-enriched fraction from nuclear proteins was obtained by affinity chromatography, and a two-dimensional gel electrophoresis （2-DE） and matrix assisted laser desorption/ionization time of matrix-assisted laser desorption/ionization/time of flight mass spectrometry （MALDI-TOF MS） were used to identify the changes of nuclear phosphoproteins in DCMECs treated with prolactin. Seven proteins displaying~〉2-fold difference in abundance upon PRL treatment in DCMECs were identified by MALDI-TOF MS. The protein-GARS （GlyRS）, which belonged to the class-II aminoacyl-tRNA synthetase family, played a global role in the milk protein synthesis. SERPINH1 （Heat shock protein 47）, which was the first heat shock protein found to be a member of the serpin superfamily, regulated physiologic functions, such as complement activation, programmed cell death, and inflammatory processes. PRDX3, which belonged to a family of antioxidant enzymes, played an important role in scavenging intracellular reactive oxygen species （ROS）. ACTR1A, belonged to the actin family, which was associated with transport of p53 to the nucleus. Annexin A2, a Ca2＋-dependent phospholipid-binding protein, maintained the viability and cell cycle regulation of DCMECs. PSMB2 and PSMD10, which belonged to ubiquitin-proteasome system, were involved in several cellular processes, including cell cycle control, cellular stress response, intracellular signaling. This screening revealed that prolactin influenced the level of nuclear phosphoproteins in DCMECs. This result opens new avenues for the study of the molecular mechanism linked to the synthesis of milk proteins.

河北省潜在赡养比解决对策之研究

潜在赡养比(The Potential Support Ratio,缩写为PSR)是伴随世界人口老龄化以及世界经济总体减缓出现的一个人口之比指标,有助于对整个经济体的整体状况予以观察。由于历史因素的造成以及经济社会发展的差异发展,河北省的潜在赡养比问题相比较更为突出。因此,加强对我省潜在赡养比问题的研究,对于理性认识河北省人口结构问题及对社会、经济等问题的研究有着积极的意义。

Role of miRNA in Mammary Gland Development and Lactation

miRNA can regulate development and milk yield of the mammary gland through epigenetic mechanism, miRNA can directly and indirectly modulate the activity of the epigenetic machinery, target genes through post-inhibition of translation initiation, mediate miRNA decay, target genes and inhibit the positive regulation, regulate tone modification, and regulate DNA methylation of target genes. Here we reviewed the role of miRNAs in mammary gland development and lactation. Researching miRNA in mammary gland development and lactation process, and understanding the response of the epigenetic mechanisms to external stimuli will be an important necessity to devise new technologies for maximizing their activity and milk production in the dairy cow.

Effect of Different Roughages on Milk Protein and Milk Fat Synthesis in Dairy Cows

The main purpose of this study was to determine the effect of a corn straw or mixed diet on milk production, milk composition and the expression of genes associated with lactation in mid-lactation Chinese Holstein cows. In this study, 10 healthy Chinese Holstein cows were randomly assigned to two groups and fed with different diets respectively, corn straw （CS） or mixed forage （MF） diet. CS group was fed roughage consisting of 53.8% corn straw only and the forge to concentrate （F ： C） ratio [dry matter （DM）] was about 40： 60. MF group was fed roughage consisting of 3.7% Chinese wildrye and 23.4% alfalfa hay, the forge to concentrate （F ： C） ratio （DM） was 70： 30. All the cows were fed 8 weeks and body weight, dry matter intake, body condition score, fat, protein, lactose, milk yield, total solid and somatic cell count （SCC） were recorded. Quantitative real-time PCR （qRT-PCR） was used to analyze cow mammary gland samples representing two different diets. The results suggested that different diet types had significant effects on milk yield, lactose, milk fat, milk protein, dry matter intake and somatic cell count in dairy cows, and cows fed MF diet improved milk production and lactation performance clearly （P〈0.05）. In addition, mRNA expression of genes ACC, roTOR, STATS, CSN2, PPAR）,, FABP3 and PTEN in MF group was extremely significantly higher than that in CS group （P〈0.05）. mRNA expression ofAKT1, FAS, SCD and SREBPlc in MF group was significantly higher than that in CS group （P〈0.01）. In summary, the milk yield and composition in mixed forage group were significantly improved than those in corn straw group.

3' Noncoding Region Construction of GHR Gene-luciferase Report Vector and Valuation

To analyze miR-139 target sites in 3＇ UTR of GHR gene in dairy cow mammary gland, a GHR 3＇ UTR- luciferase reporter vector was constructed and the effect of miRNA on its activity was evaluated in dairy cow mammary gland epithelial cells （DCMECs）. The miR-139 targeting GHR 3＇ UTR was predicted by Target Scan 5.1 software, 3＇ UTR fragment of GHR was amplified by PCR from RNA of DCMECs. PCR products were cloned into Spe Ⅰ/Hind Ⅱ modified pMIR-Report vector. The luciferase reporter vector and miRNA eukaryotic expression vector were transferred into DCMECs using lipofectamine 2000 transfection reagent. The dualluciferase reporter assay system was used to quantitiate the reporter activity. The results showed that a 107 bp 3＇ UTR fragment of GHR gene was successfully cloned into the pMIR-Report vector, which authenticated by Spe Ⅰ/Hind Ⅲ digestion and DNA sequencing. The luciferase activity of reporter construction treated with miR-139 decreased 20.87% compared with the control group. It was concluded that the GHR3＇ UTR-luciferase reporter vector had been successfully constructed. The luciferase activity of the reporter could be suppressed by miR- 139.

Potential Genes for Regulation of Milk Protein Synthesis in Dairy Goat Mammary Gland

The lactating mammary gland is a prodigious protein-producing factory, but the milk protein synthesis mechanisms are not well understood. The major objective of this paper was to elucidate which genes and pathways were involved in the regulation of milk protein synthesis in the dairy goat mammary gland. Total 36 primiparous Guanzhong dairy goats were allotted in 12 groups according to their mammary development stages： days 90 and 150 of virgin, days 30, 90, and 150 of pregnancy, days 1, 10, 35, and 60 of lactation and days 3, 7, and 21 of involution （three animals per group）. Mammary tissue RNA was isolated for quantitative real- time RT-PCR of four casein genes alpha-s 1 casein （CSN 1S 1 ）, alpha-s2 casein （CSN 1 S2）, beta-casein （CSN2） and casein kappa （CSN3）, four whey protein genes lactoglobulin （LGB）, laetalbumin （LALBA）, laetofarrin （LTF）, and Whey acidic protein （WAP） and the genes which were potentially to regulate dairy goat milk protein synthesis at the level of transcription or translation [prolactin receptor （PRLR）, AKT1, signal transducers and activators of transcription 5 （STAT5）, E74-Like Factor 5 （ELF5）, eukaryotic translation initiation factor 4E binding protein 1 （EIF4E-BP1）, S6kinase （S6K） and caveolin 1]. The results showed that all genes were up-regulated in lactation period. The expressions of PRLR, AKT1, STAT5, ELF5, and S6K were similar to mRNA expressions of milk proteins. Our results indicated that milk protein synthesis in dairy goat mammary gland was possibly regulated by these genes.

河北省优秀传统文化的近期偏好、现代阐释和愿景设计

河北省作为京津冀协同发展及其雄安新区建设的重要一环,具有很强文化资源禀赋。通过对河北省传统文化近期偏好、现代阐释和愿景设计的研究,为河北省消除地区文化差异和经济差异提供理论指导,并为之注入现代内涵,具有很强的现实意义和理论意义。

京津冀社会保障体系一体化的历史借鉴及对策

京津冀协同发展战略为三省市的社会保障一体化提供了难得的机遇,在对三省市的社会保障一体化的历史借鉴基础上,必须重视法律建设、顶层设计和基本公共服务均等化的协同发展。

环渤海经济区生态环境协同治理的法治化研究

环渤海经济区的生态环境协同治理是一个战略性的问题,关涉政治、经济、文化等多重因素。必须在考虑有关各方的利益下,通过法治的手段,建立相应的合作与救助机制,实现环渤海生态环境协同治理的可持续发展。