Effects of Energy,Protein and Amino Acids Levels on the Development of Mammary Gland and the Growth of Fetus in Sows at Late Gestation

Late gestation is an important period for rapid development of the sow's mammary gland and fetus. It is becoming more and more important to provide a suitable nutrient level in late gestation period for improve the reproductive efficiency of sow. Demand of energy, protein and amino acids for mammary gland and fetus in sows at late gestation are different. To provide reasonable nutrition for fetus, at the same time for the needs of mammary gland are still seldom reported. In this paper, the effects of dietary energy, protein and amino acids levels on the development of mammary gland and the growth of fetus were reviewed,the authors propose the requirement of energy, protein amino acid and reasonable proportion in sows at late gestation, in order to give some guidance for pig farmers.     

脂肪酸对奶牛乳腺上皮细胞乳蛋白和乳脂合成相关基因表达量的影响

本试验旨在探寻促进奶牛乳腺上皮细胞(BMECs)乳蛋白和乳脂合成的短链脂肪酸(乙酸、β-羟丁酸)和长链脂肪酸(油酸、亚油酸、亚麻酸)的组合添加模式,为调控乳成分合成提供理论依据。BMECs经分离、纯化后,选取第2代细胞,分为5组,对照组不添加脂肪酸,Ⅰ组和Ⅱ组添加的乙酸、β-羟丁酸浓度比例均为2.0(9.60 mmol/L)∶1.0(4.80 mmol/L),油酸、亚油酸、亚麻酸的浓度比例分别为2.0(17.30μmol/L)∶13.3(115.05μmol/L)∶1.0(8.65μmol/L)和9.6(75.20μmol/L)∶7.4(58.00μmol/L)∶1.0(7.80μmol/L);Ⅲ组和Ⅳ组添加的乙酸、β-羟丁酸的浓度比例均为1.0(7.20 mmol/L)∶1.0(7.20 mmol/L),油酸、亚油酸、亚麻酸的浓度比例分别为2.0∶13.3∶1.0和9.6∶7.4∶1.0,各组添加的短链脂肪酸(SCFA)和长链脂肪酸(LCFA)总浓度为14.541 mmol/L,每组3个重复。培养24 h后,检测细胞相对增殖率(RGR)、甘油三酯(TAG)的合成量以及乳蛋白和乳脂合成相关基因的表达量。结果表明:1)试验组BMECs RGR及TAG的合成量均显著高于对照组(P0.05);Ⅰ组RGR最高,TAG合成量最多。2)与对照组相比,Ⅱ组显著提高了核糖体蛋白S6激酶1(S6K1)、κ-酪蛋白(CSN3)基因的表达量(P0.05);Ⅳ组显著提高了CSN3、蛋白激酶B(AKT)、S6K1、真核翻译起始因子4E结合蛋白1(4EBP1)基因的表达量(P0.05);试验组信号转导和转录激活因子5(STAT5)基因的表达量显著降低(P0.05)。3)与对照组相比,试验组二酰甘油酰基转移酶2(DG AT2)基因的表达量显著提高(P0.05),脂肪酸合成酶(FASN)基因的表达量显著降低(P0.05)。综上所述,在培养液中添加7.20 mmol/L乙酸、7.20 mmol/Lβ-羟丁酸、75.20μmol/L油酸、58.00μmol/L亚油酸、7.80μmol/L亚麻酸对BM ECs乳蛋白和乳脂合成相关基因的表达量有较好的促进作用。     

Expression of E‐cadherin in canine anal sac gland carcinoma and its association with survival*

The objective of this study was to determine whether an association could be demonstrated between survival and the expression of the adhesion molecule E‐cadherin by the neoplastic cells in a group of dogs with anal sac gland carcinomas (ASGCs). Archived formalin‐fixed, paraffin wax‐embedded primary tumour specimens were obtained for 36 cases of canine ASGC with known clinical management and survival data. Immunohistochemical methods were used to evaluate E‐cadherin expression by the neoplastic cells and data were evaluated for an association between E‐cadherin expression and survival. On univariate analysis, the median survival time for cases with tumours expressing E‐cadherin in more than 75% of cells was significantly greater than that for cases with tumours expressing E‐cadherin in fewer than 75% of cells (1168 versus 448 days, P = 0.0246). Both E‐cadherin expression and presence or absence of distant metastases were significantly associated with survival on multivariate analysis. This study demonstrates that expression of E‐cadherin at the cytoplasmic membrane in canine ASGCs is variable and potentially predictive of survival.     

东北虎主要消化腺的组织学研究

通过光镜对东北虎主要几种消化腺的H.E染色石蜡切片进行观察,结果表明,东北虎肝小叶间的结缔组织不发达,小叶的界限不清晰,肝细胞体积较大,核位于细胞一侧,肝细胞多呈索状排列,窦状隙不明显;胰腺外分泌部由腺泡和导管组成,腺泡形状不一,由单层锥状腺上皮细胞组成,胰岛是一些大小不等的细胞团和细胞索,分散于外分泌部之间,胞质染色较外分泌部浅;腮腺为纯浆液腺泡,胞质着色呈嗜酸性,颌下腺是以黏液性细胞为主,含少量的浆液性细胞半月。     

Characterization of spheres derived from canine mammary gland adenocarcinoma cell lines

There is increasing evidence for the presence of cancer stem cells in several solid tumors, and these cancer stem cells have a potential role in tumor initiation, aggression, and recurrence. The stem cell-like properties of spheres derived from canine mammary tumors remain largely elusive. We attempted to induce sphere formation using four cell lines of canine mammary adenocarcinoma, and characterized the spheres derived from a CHMp line in vitro and in vivo. The CHMp-derived spheres showed predominantly CD44⁺CD24⁻ population, higher expression of stem cell-related genes, such as CD133, Notch3 and MDR, and higher resistance to doxorubicin compared with the CHMp-derived adherent cells. Xenograft transplantations in nude mice demonstrated that only 1 × 10⁴ sphere cells were sufficient for tumor formation. Use of the sphere assay on these sphere-derived tumors showed that sphere-forming cells were present in the tumors, and were maintained in serial transplantation. We propose that spheres derived from canine mammary adenocarcinoma cell lines possess a potential characteristic of cancer stem cells. Spheres derived from canine mammary tumors could be a powerful tool with which to investigate novel therapeutic drugs and to elucidate the molecular and cellular mechanisms that underlie tumorigenesis.     

Identification of Suspected Stem/progenitor Cells in Bovine Mammary Epithelial Cells Culture System

To develop the potential function of dairy cow mammary stem cells (DCMECs) in regulation of lactation,we identify putative DCMECs which were BrdU label retaining epithelial cells,at the same time,analysis the location of two new mammary stem cells molecular marks FNDC3B and PROCR to verify the feasibility of them to indicate DCMECs.The mRNA levels of prolactin,growth hormone,insulin-like growth factor-1 and their receptors were detected along with cell passage by Real-time quantitative PCR.The results showed that the proportion of BrdU label-retaining epithelial cells was nearly 0.4% after 25 d continuous culture (passaged 4 times) and few cells were positive for FNDC3B or PROCR.Moreover,we observed the BrdU labelled epithelial cells by asymmetric division.The mRNA levels of prolactin,growth hormone,insulin-like growth factor-Ⅰ and their receptors in primary and passage cells were extremely significant difference(P<0.01).DCMECs would rapidly lose some physiological characteristics and the ability of milk synthesis when not under the condition of induction of lactation differentiation,but a certain percentage of mammary stem/progenitor cells will be retained,whose potential effects on the regulation of lactation and mammary acinar remodeling were worthy of attention.     

Effect Comparison of Adenovirus Vector Transfer into Different Buffalo Cells

Buffalo mammary epithelial cell,cumulus cell and fibroblasts were transfected by adenovirus vectors and compared their transfection efficiency.293 cells were transfected with pBHGloxdelE13cre and pDC316-eGFP by liposome,the virus was collected and titer was detected.Buffalo mammary epithelial cell,cumulus cell and fibroblasts were exposed to different multiplicity of infection (MOI) of adenovirus vectors.After 72 h,the cells were observed with inverted fluorescence microscope,and transfection efficiency was calculated.When the MOI was 25,50,100,200 and 400,the transfection efficiency of fibroblasts were 0.7%,7.0%,9.0%,12.5% and 34.0%,the transfection efficiency of cumulus cells were 42.5%,55.3%,57.4%,76.0% and 80.0%,the transfection efficiency of mammary epithelial cells were 88.7%,100%,100%,100% and 100%.The results showed that the transfection efficiency of mammary epithelial cell was the best,followed by cumulus cell,and fibroblast was poor.     

金黄色葡萄球菌对奶牛乳腺上皮细胞Wnt/β-catenin信号通路和相关修复因子的影响

采用金黄色葡萄球菌侵袭原代奶牛乳腺上皮细胞,观察该病原菌对乳腺上皮细胞Wnt/β-catenin信号通路关键蛋白和相关修复因子EGFR、TGF-β3和VEGF基因mRNA表达的影响。金黄色葡萄球菌以MOI=1∶1的比例接种奶牛乳腺上皮细胞,分别作用0,15,30,45,60,120,240 min,采用Western blot法检测乳腺上皮细胞β-catenin、Cyclin D1及c-Myc蛋白表达水平;免疫荧光法检测β-catenin的表达及核易位;荧光定量PCR检测EGFR、TGF-β3和VEGF基因mRNA表达。结果显示,β-catenin蛋白在45,60,120 min表达量升高,与0 min相比差异显著(P<0.05);Cyclin D1蛋白在45,60,120和240 min表达量升高,差异显著或极显著(P<0.05或P<0.01);c-Myc蛋白在15,30,60,120,240 min表达量升高,差异极显著(P<0.01)。修复相关因子EGFR、TGF-β3和VEGF基因mRNA的表达量在30 min均出现极显著升高(P<0.01),且VEGF基因mRNA在45,60,120和240 min均呈现极显著升高(P<0.01),EGFR基因mRNA表达量在30,45和60 min时表达量极显著升高(P<0.01)。结果表明,金黄色葡萄球菌侵袭奶牛乳腺上皮细胞后,诱导Wnt/β-catenin信号通路的转导和修复因子EGFR、TGF-β3和VEGF基因转录,该信号通路和修复因子可能参与金黄色葡萄球菌导致的炎症和细胞损伤的修复过程。