Astrocyte proliferation in the rat hippocampus after middle cerebral artery occlusion

AIM: To study rat astrocyte proliferation in ipsilateral hippocampus following focal cerebral ischemia. METHODS: Ischemia was induced by temporary middle cerebral artery occlusion (MCAO). In hippocampus of rats at 3, 7 and 30 days after MCAO, the numbers and anatomic distribution of glial fibrillary acidic protein (GFAP) were detected by immunohistochemistry. The protein expression of GFAP and proliferating cell nuclear antigen (PCNA) in the ipsilateral hippocampus were analyzed by Western blot analysis. RESULTS: Astrocytes appeared hypertrophic, with increased process thickness and numbers at 7 days after MCAO, and the highest density of astrocytes were seen at 30 days in the CA1, CA2 regions of the ipsilateral hippocampus. Western blot analysis revealed that GFAP levels were normal at 3 days, but increased by 7 days and remained elevation at 30 days. Western blot analysis of PCNA protein also revealed identified upregulation PCNA at 3 days after MCAO and the expression peaked at 7 days. CONCLUSION: This study demonstrates that focal cerebral ischemia in the rat results in a rapid response, a process often referred to as reactive astrogliosis or glial scarring, from resident astrocytes of the ipsilateral hippocampus to the side of ischemia.     

Meiotic competence of mouse oocytes reconstructed by replacing the germinal vesicle with a male pronucleus and somatic nucleus

The present study examines the contribution of the nucleus to meiotic competence in mouse oocytes that were reconstructed using nuclear transfer. Three types of reconstructed oocytes were produced: MP‐GV, by transplanting the male pronucleus (MP) into germinal vesicle (GV) stage oocytes; 3T3‐GV, by transplanting the nucleus of a National Institute of Health (NIH) 3T3 cell into a GV stage oocyte; and 3T3‐MII, by transplanting the nucleus of an NIH 3T3 cell into a metaphase II (MII) stage oocyte. The fusion rates differed, but not significantly, in the MP‐GV, 3T3‐GV, and 3T3‐MII groups (77, 63, 56%, respectively). Then, meiotic competence was compared in MP‐GV, 3T3‐GV and non‐manipulated GV stage oocytes as a control. Nuclear envelope breakdown occurred in all the reconstructed oocytes, as well as the control ones. The percentage of first polar body extrusion differed between the MP‐GV (100%), 3T3‐GV (72%), and control (67%) groups. DNA staining with Hoechst 33342 revealed that in the MP‐GV‐group oocytes that had reached MII stage, the chromosomes were condensed and aligned in a regular array similar to the normal metaphase plate. By contrast, in 3T3‐GV group oocytes, the condensed chromosomes were irregularly scattered in the cytoplasm. These results suggest that the donor nucleus affects meiotic competence in reconstructed oocytes.     

核磁共振法测定甲哌鎓原药的含量

采用定量核磁共振法建立了甲哌鎓原药的定量方法。以重水为溶剂,仲丁基脲为内标物,以化学位移分别在δ3.12的甲哌鎓甲基和δ1.10的仲丁基脲甲基的1H-NMR信号作为定量峰。以甲哌鎓与仲丁基脲定量峰的峰面积比(AS/AI)对甲哌鎓与仲丁基脲的质量比(WS/WI)进行线性回归,线性相关系数为0.999 9;重复测定同一批次原药含量,变异系数为0.28%。采用核磁共振波谱法对甲哌鎓进行定量分析,操作简便,测定速度快,准确度高。     

瘤胃液移植在反刍动物中的研究进展

瘤胃液移植是一种将健康供体动物的瘤胃液转移给受体动物的技术。目前，针对瘤胃液移植的报道还相对有限，但越来越多的研究结果显示，瘤胃液移植可以影响反刍动物的干物质采食量、产奶量等，还具有重建瘤胃菌群结构的潜力。本文综述了瘤胃液移植的方法及实践中的应用研究，分析其对反刍动物生长性能、瘤胃发酵、瘤胃微生物区系的影响以及在治疗某些疾病方面的作用，为反刍动物瘤胃液移植的研究提供参考。     

不同合成树脂工艺对改性材性能影响研究

为研究树脂对改性材性能的影响，采用2种不同工艺合成三聚氰胺-脲醛树脂（MUF），测试了树脂的相关性能。结果表明，不同合成工艺路线下制备的 MUF 树脂在固体含量、粘度、固化时间、游离甲醛含量间存在显著差异。最终树脂的分子结构类型相似性极高，但相同结构组分在不同树脂中所占比例各有差异。羟甲基基团在MUF2中所占比例高，而亚甲基桥键及醚键在MUF1中含量高。MUF1改性材的增重率（weight percent gain，WPG）值更大，但MUF2改性材的抗溶胀性（anti-swelling efficiency，ASE）和体积膨胀率（bulking rates，B）更高，MUF2改性材的尺寸稳定性更好。     

GMP玻璃化冷冻未成熟牛卵母细胞核成熟及冷冻损伤试验

本试验通过荧光染色的方法建立了未成熟牛卵母细胞在体外培养过程中第1次减数分裂的各个阶段的参考判定图谱;根据这个标准来观察毛细玻璃管(GMP)玻璃化冷冻对卵母细胞核成熟和冷冻损伤的影响。结果表明,从屠宰场废弃的卵巢表面卵泡内抽取的COCs,70%处于生发泡期,12.5%生发泡开始破裂,7.5%已开始浓缩,这说明从屠宰场获得的COCs有较高的异质性;卵母细胞在成熟培养22h时收获排出第一极体的卵母细胞,可得到丰度较高的极体-胞质染色体对称、紧密相邻的成熟卵母细胞;GMP玻璃化冷冻损伤主要有2种表现形式,首先,直接影响膜结构的完整性,包括细胞膜和核膜,这可从退化的细胞比例看出(8～24h,有21.9%～27.2%的细胞处于该阶段),其次,影响CONDENSED向MⅠ期的过渡,这可从处于CONDENSED卵母细胞的比例看出(8～24h,有24.1%～34.3%的细胞处于该阶段)。     

Production and Identification of Clone Sheep Using Bone Marrow Mesenchymal Stem Cells as Donor Cell

In order to obtain the cloned sheep using bone marrow mesenchymal stem cells（BMSCs）as the donor cells,the BMSCs of sheep were chosen and reconstructed embryos were built to transfer.10 published microsatellite markers were chosen,and the DNA samples from clone sheep,donor cells and surrogate ewes were amplified,and the relationship of father-son（RCP）was analyzed using the Quantity One for genotyping.The results showed that the reconstructed embryos were successfully built for electric fusion using sheep BMSCs as nuclear donor,and making nuclear transplantation into enucleated mature oocytes of which the fusion rate was 80.62%.20 surrogate ewe were chosen to be implanted with the reconstructed embryos at morula stage by implant surgery,and 5 lambs were born and only 3 were survived.The genotype of cloned sheep was in line with the dornor cell and the RCP were more than 99.999%.In conclusion,the first clone sheep were obtained successfully by using BMSCs as a nuclear donor in this experiment.     

Improved development of somatic cell cloned bovine embryos by a mammary gland epithelia cells in vitro model

Previous studies have established a bovine mammary gland epithelia cells in vitro model by the adenovirus-mediated telomerase (hTERT-bMGEs). The present study was conducted to confirm whether hTERT-bMGEs were effective target cells to improve the efficiency of transgenic expression and somatic cell nuclear transfer (SCNT). To accomplish this, a mammary-specific vector encoding human lysozyme and green fluorescent protein was used to verify the transgenic efficiency of hTERT-bMGEs, and untreated bovine mammary gland epithelial cells (bMGEs) were used as a control group. The results showed that the hTERT-bMGEs group had much higher transgenic efficiency and protein expression than the bMGEs group. Furthermore, the nontransgenic and transgenic hTERT-bMGEs were used as donor cells to evaluate the efficiency of SCNT. There were no significant differences in rates of cleavage or blastocysts or hatched blastocysts of cloned embryos from nontransgenic hTERT-bMGEs at passage 18 and 28 groups (82.8% vs. 81.9%, 28.6% vs. 24.8%, 58.6% vs. 55.3%, respectively) and the transgenic group (80.8%, 26.5% and 53.4%); however, they were significantly higher than the bMGEs group (71.2%, 12.8% and 14.8%), (p < 0.05). We confirmed that hTERT-bMGEs could serve as effective target cells for improving development of somatic cell cloned cattle embryos.     

软枣猕猴桃试管苗驯化移栽技术的研究

[目的]探索软枣猕猴桃试管苗移栽后的最宜生长条件,提高成活率,缩短成活时间。[方法]通过测试不同炼苗季节、预处理方法、栽培基质、温度、湿度、光照对试管苗成活、移栽效果影响,得出最佳移栽炼苗程序。[结果]将生长健壮,苗高5cm以上的组培苗移至室温光照2000-3000lx,10h/d的栽培条件下经4d炼苗后,清水洗苗,蘸生根粉移栽至沙土∶黄棕壤∶腐叶土∶园土(1∶1∶1∶1)的基质中盆栽、套袋后置于棚高50cm的小拱棚内,拉2层遮阴网（透光率40-50%,温度20-24℃,土壤含水量80-90%）培养出的幼苗,成活率高达92%,叶数7,整体生长整齐,是软枣猕猴桃最佳驯化移栽方式。[结论]实验所得方法可为大田生产提供基础,极大程度上提高移栽成活率,缩短成活时间。     

植物增殖细胞核抗原与DNA复制关系的研究现状

为了更深入的对植物增殖细胞核抗原(proliferating cell nuclear antigen, PCNA)进行研究,本研究归纳了PCNA的研究现状,总结了PCNA的结构特征和包括PCNA参与DNA复制、DNA损伤修复和细胞周期调控等在内的PCNA的功能特点。目前有关PCNA的研究以医学和动物体为主,有关植物PCNA研究的报道很少且相对落后,但已有文献指出,PCNA调控DNA复制,参与DNA复制以确保植物体正常生长发育,因此植物PCNA的功能值得继续研究。