Effects of Confluent, Roscovitine Treatment and Serum Starvation on the Cell-cycle Synchronization of Bovine Foetal Fibroblasts

The present study was designed to examine the effects of cell-cycle synchronization protocols, such as confluent, roscovitine treatment and serum starvation, in bovine foetal fibroblasts on synchronization accuracy at G0/G1, viability, apoptosis, necrosis and ploidy for use as a nuclei donor. The cells in 5-10 passages were randomly allocated into three treated groups. Cells were cultured either in Dulbecco's modified Eagle's medium (DMEM) + 10% foetal bovine serum (FBS) until 90% confluent (group 1, confluent), in DMEM + 10% FBS + 30 microM roscovitine for 12 h (group 2, roscovitine), or in DMEM + 0.5% FBS for 5 days (group 3, serum starvation). Most of the cells (>80%) in all groups were arrested at the G0/G1 stage. Although the rates did not differ, cells in group 1 showed an increased cell population arrested at the G0/G1 phase. Significantly (p < 0.05) higher rates of apoptosis occurred in group 3 than in group 1 and 2 (10% vs 6% and 6%, respectively). No differences in chromosomal abnormality were observed among groups. However, by increasing the number of cell culture passages up to 15, significantly (p < 0.05) higher chromosomal abnormality was observed than in 5 and 10 passages (39% vs 28% and 23%, respectively) in group 1. The results clearly indicated that bovine foetal fibroblasts could be effectively synchronized at G0/G1 stages by all the three different treatments, confluent, roscovitine and serum starvation. However, cells in confluent showed reduced apoptosis and necrosis when they underwent 5-10 passages, exhibiting increased percentage of cells with stable chromosome diversity. Hence, cells in confluent merit further studies before they could be used as nuclear donors.     

“珍珠”的工艺配方研究

通过正交试验,对"珍珠"的工艺配方进行了研究,结果表明,胶液最佳配方为海藻酸钠∶明胶∶琼脂为3∶1∶2,奶油含量5%,蔗糖含量10%。氯化钙浓度3.0%,固化时间10min。得到的"珍珠"颗粒形状圆润,口感滑嫩,3个月内不发生溶解。     

长爪沙鼠颌下腺免疫球蛋白A的定位分布

采用免疫组织化学方法对不同日龄的长爪沙鼠颌下腺IgA的定位分布进行了研究。结果表明，长爪沙鼠的颌下腺由导管部和分泌部构成，分泌部主要由浆液腺构成，导管部包括闰管、纹管、颗粒曲管和小叶间导管等。DAB显色结果表明，IgA阳性细胞主要分布于浆液性腺泡、闰管、纹管、颗粒曲管和小叶间导管，并可分布于腺泡和腺管间结缔组织，IgA阳性产物的分布具有不均一性，无明显随年龄变化的规律性。阳性产物分布于胞质中，胞核呈阴性，对照组阴性。提示从浆细胞产生或循环而来的IgA先经结缔组织进入颌下腺组织，进而定位分布于浆液腺泡和各级导管，导管部有较多的IgA分布。     

The Control of Reactive Oxygen Species Influences Porcine Oocyte In Vitro Maturation

The aim of this study was to examine the effect of varying intracellular reactive oxygen species (ROS) levels during oocyte in vitro maturation with enzymatic ROS production systems (xanthine + xanthine oxidase or xanthine + xanthine oxidase + catalase), scavenger systems (catalase or superoxide dismutase + catalase) or cysteine on porcine oocyte maturation. Oocyte ROS levels showed an increase when H₂O₂ or O₂?^‐ production systems were added to the culture medium (p < 0.05). On the other hand, the presence of ROS scavengers in the maturation medium did not modify oocyte ROS levels compared with the control after 48 h of maturation, but the addition of cysteine induced a decrease in oocyte ROS levels (p < 0.05). The ROS production systems used in this work did not modified the percentage of oocyte nuclear maturation, but increased the decondensation of sperm head (p < 0.05) and decreased the pronuclear formation (p < 0.05). In turn, the addition of O₂?^‐ and H₂O₂ scavenging systems during in vitro maturation did not modify the percentage of oocytes reaching metaphase II nor the oocytes with decondensed sperm head or pronuclei after fertilization. However, both parameters increased in the presence of cysteine (p < 0.05). The exogenous generation of O₂?^‐ and H₂O₂ during oocyte in vitro maturation would not affect nuclear maturation or later sperm penetration, but most of the spermatozoa cannot progress to form the pronuclei after fusion with the oocyte. The decrease in endogenous ROS levels by the addition of cysteine would improve pronuclear formation after sperm penetration.     

Determination of reference intervals for metabolic profile of Hanwoo cows at early,middle and late gestation periods

Background

Metabolic profile was initially designed as a presymptomatic diagnostic aid based on statistical analyses of blood metabolites to provide an early warning of certain types of metabolic disorder. However, there is little metabolic profile data available about Korean Hanwoo cows. Therefore, this study aimed to determine the reference intervals of metabolic profile for Korean Hanwoo cows.

Methods

Healthy animals (2,205) were selected and divided into early (day 1 to 95), middle (day 96 to 190) and late (day 191 to 285) period according to their gestating period. Metabolic profile including total protein (TP), albumin (Alb), urea (UREA), glucose (Glu), total cholesterol (T-Cho), long-chain fatty acid (LCFA), aspartate aminotransferase (AST), gamma-glutamyl transpeptidase (GGT), creatinine (Crea), calcium (Ca), inorganic phosphorous (iP) and magnesium (Mg) were analyzed using a TBA-40FR automatic biochemical analyzer. The data of Korean Hanwoo cows were then compared to those of the Japanese Wagyu cows.

Results

Most of the data of the Korean Hanwoo cows were relatively higher than those of Japanese Wagyu cows, with the exception of Glu and GGT. This may indicate that the nutritional level of feed for the Korean Hanwoo cows was higher than that of the Japanese Wagyu cows because of the different feeding system. In particular, relatively higher levels of UREA and LCFA were observed in the Korean Hanwoo cows, and this may also contribute to the low reproduction efficiency.

Conclusions

These findings may provide some theoretical basis for understanding the reproductive and feeding situation of Korean Hanwoo cows.     

城市微型公共空间景观设计

随着城市的发展,城市公共空间对城市生活的重要性不断凸显,城市微型公共空间作为其层次结构中不可或缺的一环,在世界范围内也得到了长足发展。但对于我国城市微型公共空间景观设计领域而言,却一直未能得到足够的重视。以深圳市罗湖区嘉宾公园为研究对象,阐述城市微型公共景观设计对于城市公共空间层次结构的重要意义,并展现其为城市所带来的独特魅力。深圳市罗胡区嘉实公园的建成为我国城市微型公共空间提供了一个极佳的参考样本。