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.     

秀丽纤杆线虫在寄生性线虫抗原研究中的应用

秀丽纤杆线虫 (Caenorhabditis elegans)是一种模式线虫 ,目前广泛应用于寄生性线虫基因的功能研究 ,并且还被用于捻转血矛线虫等寄生性线虫疫苗侯选抗原基因的表达和耐药性产生机制方面的研究     

禽大肠杆菌外膜蛋白基因C(ompC)的序列分析

根据 Gen Bank中人源大肠杆菌 K- 12外膜蛋白基因 C(omp C)的核苷酸序列设计引物 ,应用 PCR方法从禽大肠杆菌 O2 、O78株及它们的融合双价弱毒菌株 O2 ,78(Norr Chlr)中分别扩增得到 omp C基因 ,序列测定及分析比较发现 ,3个菌株的 om p C基因均由 170 2 nt组成 ,核苷酸序列完全相同 ,只有 1个大的开放性阅读框 (ORF) ,长 10 92 bp,编码由 36 3个氨基酸组成的前 Om p C蛋白 ,前 2 1个氨基酸残基组成信号肽 ,成熟的 Omp C蛋白由 342个氨基酸残基组成 ,Mr为 4 0 0 0 0。其氨基酸序列也完全相同。从基因水平上证明了禽大肠杆菌 O2 、O78株及融合双价弱毒菌株 O2 ,78(NorrChlr)存在相同的外膜蛋白 C抗原 ,从而为进一步研究 Omp C蛋白的免疫原性奠定了基础     

羊、鸡抗鼠脂肪细胞膜抗血清对大鼠体脂与生长性能的影响

应用提取的鼠脂肪细胞膜分别免疫羊和鸡 ,所产生的抗血清用于 Wistar大鼠被动免疫。实验 1: 组腹腔注射羊正常血清 , 组腹腔注射羊抗鼠脂肪细胞膜抗血清 ,剂量均为 1m L /只 ,连续注射 4 d。结果表明 ,羊抗鼠脂肪细胞膜抗血清免疫促进了大鼠体增重 ,降低了体脂沉积 ,与对照组相比 ,7周末体重增加 6 .35 % (P<0 .0 5 ) ,饲料摄入增加6 .85 % (P<0 .0 1) ,料重比 (F/G)提高 4 5 .0 0 % (P<0 .0 5 ) ;肾周、附睾、网膜脂肪垫重量分别降低 2 3.92 % (P<0 .0 5 )、34.4 5 % (P<0 .0 5 )、0 .98% ,脂肪总量降低 2 0 .92 %。实验 2 :1组腹腔注射鸡正常血清 ,2组腹腔注射鸡抗鼠脂肪细胞膜抗血清 ,剂量均为 1m L/只 ,连续注射 4 d。结果表明 ,鸡抗鼠脂肪细胞膜抗血清免疫对大鼠的生长发育产生了不利影响 ,7周末 ,免疫大鼠平均体重较对照组减少 4 0 g(P<0 .0 5 ) ,饲料摄入显著降低 (P<0 .0 1) ;对体脂的沉积和血液中 TG和 FFA的影响没有规律 ,且无统计学意义     

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.     

Disaccharide combinations and the expression of enolase3 and plasma membrane Ca2+ ATPase isoform in sturgeon sperm cryopreservation

Acipenser sinensis and Acipenser dabryanus are critically endangered species, so germplasm conservation via cryopreservation of sperm is necessary. Disaccharides can act as membrane‐impermeable cryoprotectants, and enolase3 (ENO3) and plasma membrane Ca²⁺ ATPase isoform (PMCA2) are proteins associated with sperm quality. We considered seven characteristics of sperm quality in cultured brood stock from A. sinensis and A. dabryanus. We tested use of sucrose or trehalose alone and in combination at different concentrations for cryopreservation of A. dabryanus sperm. A low concentration of sucrose plus trehalose (S₁₅T₁₅) was optimal. Mixing of the extender with sucrose, lactose, or trehalose alone or with pairwise mixtures revealed that a mixture of lactose and trehalose (L₁₅T₁₅) gave the best results for both A. sinensis and A. dabryanus. Enolase3 and PMCA2 expression levels were measured in cryopreserved A. sinensis sperm via Western blotting. Relative ENO3 and PMCA2 expression levels were examined, and the relationship between disaccharide composition, sperm quality and protein expression was explored in A. sinensis. The results showed that relative ENO3 and PMCA2 expression levels were the highest at L₁₅T₁₅ in cryopreserved A. sinensis sperm. There were significant positive correlations between ENO3 expression and percentage membrane integrity, and between PMCA2 expression and sperm motility parameters (percentage of motile sperm, curvilinear velocity, straight‐line velocity and average path velocity; p < .05) in cryopreserved A. sinensis sperm. Our results indicate the optimal disaccharide combination and concentrations for cryopreservation of A. sinensis and A. dabryanus sperm and suggest that ENO3 and PMCA2 expression levels could serve as a valuable indicator of sperm quality in A. sinensis.     

Accessory nictitating membrane in a Lipizzaner colt

A 2‐year‐old Lipizzaner colt presented for removal of a mass over the left eye. The colt had no blepharospasm or significant ocular discharge, but the cornea underlying the mass was mildly oedematous. The mass protruded from the conjunctiva at the dorsotemporal aspect of the globe and was covered in normal conjunctiva. It had a gross appearance similar to the cartilaginous flap of a nictitating membrane, but in an aberrant location. It was moveable to cover the dorsal aspect of the cornea when the globe was retropulsed. Excision of the mass under general anaesthesia was elected. Histopathology supported a diagnosis of an accessory nictitating membrane, consisting of a cartilaginous band surrounded by glandular epithelium. The colt recovered from surgery without complication and no problems were reported by the owner at 8.5 months post‐operatively. To the author's knowledge, there have not been previous reports of accessory or ectopic nictitating membranes in equine or other species.     

携带猪流行性腹泻病毒S1抗原表位的乙肝核心抗原病毒样颗粒的构建与鉴定

旨在构建以乙肝核心抗原(HBcAg)为载体呈现猪流行性腹泻病毒(PEDV)S1抗原表位的病毒样颗粒.将PEDV S1蛋白中含B细胞表位的270 bp片段插入到HBcAg主要免疫显性区域(MIR),构建重组质粒pET-32a(+)-HBcAg-PEDV S1,转化到感受态细胞BL21(DE3)中,经IPTG诱导表达,SD...     

木聚糖酶在膜反应器内的酶解特性

分别研究了粗木聚糖酶和纯化的木聚糖酶在超滤膜反应器(UMR)和常规反应器(CSBR)中的酶解特性。粗木聚糖酶或纯木聚糖酶在UMR中酶解木聚糖时,反应进行了525 m in时所得产品中低聚木糖各组分的质量分数(木二糖~木五糖)均在20%左右,木糖质量分数约为9.5%。在UMR中粗木聚糖酶降解木聚糖时的低聚木糖得率、低聚木糖占总糖的比例和低聚木糖生产能力比纯木聚糖酶在CSBR中分别高19.1%、14.8%和13.5%;而木糖的得率却低55.2%。粗木聚糖酶在UMR中酶解木聚糖时,所得低聚木糖产品中木二糖~木五糖组分含量基本相等;纯木聚糖酶在CSBR中酶解木聚糖时,所得低聚木糖产品中木二糖含量较高。同纯木聚糖酶在CSBR中酶解特性相比,粗木聚糖酶在UMR中酶解木聚糖可以制得高质量低聚木糖。