转EGFP基因山羊克隆胚的发育     

郑月茂  刘凤军  安志兴  李向臣  赵晓娥  权富生  张涌 《畜牧兽医学报》2007,38(5):458-463

利用脂质体包裹含EGFP基因的质粒,并将之导入山羊乳腺上皮细胞,经G418筛选获得阳性细胞,以阳性细胞作为核供体,利用核移植技术构建转基因克隆胚。结果表明：用转基因山羊乳腺上皮细胞作为核供体,电融合法更适合构建转基因克隆胚。转基因山羊克隆胚体外培养最佳方案是用SOFaa培养液,在培养72 h后加入10%的正常山羊血清（Normal goat serum,NGS）。荧光显微镜下观察到转基因克隆胚中EGFP的表达,大部分克隆胚发育到8-16细胞以后的时期,绿色荧光蛋白才开始逐渐表达,随着发育时间的延长,绿色荧光蛋白的表达也逐渐增强。说明外源基因在胚胎早期发育阶段可以表达,EGFP可以作为报告基因来实现对外源基因整合及表达的监测。  相似文献   

Stimulation of the bronchus-associated lymphoid tissue of goats and its effect on in vitro colonization by Pasteurella haemolytica     

Effendy  A.W.M.  Zamri-Saad  M.  Maswati  M.A.  Ismail  M.S.  Jamil  S.M. 《Veterinary research communications》1998,22(3):147-153

Twenty goats of about 7 months of age were divided into five groups. The goats in groups 1 and 2 were exposed once, using an intranasal spray to 2 ml of an inoculum containing 10⁶ colony-forming units/ml of living or dead Pasteurella haemolytica A2, respectively. The goats in groups 3 and 4 were similarly exposed twice at a 2-week interval. Group 5 was the untreated control. The number and size of the bronchus-associated lymphoid tissue (BALT) in goats exposed twice to either living or dead organisms were significantly (p>0.05) increased compared with those exposed once and with the unexposed control. In vitro colonization by living P. haemolytica A2 onto the lung tissue in which the BALT had been stimulated by two exposures of either living or dead organisms was significantly (p>0.05) reduced. The study indicates that stimulation of the respiratory mucosal immunity may prevent P. haemolytica A2 infection.  相似文献   

ULTRASONOGRAPHIC DETECTION OF ADRENAL GLAND TUMOR AND URETEROLITHIASIS IN A GUINEA PIG   总被引：1，自引：0，他引：1  

Lorrie  Gaschen  DVM  Cornelia  Ketz  Dr. med. vet.  Johann  Lang  Dr. med. vet  Urs  Weber  Dr. med. vet.  L. Bacciarini  Dr. med. vet.  Ingrid  Kohler  med. vet. 《Veterinary radiology & ultrasound》1998,39(1):43-46

A 5-year-old guinea pig was presented to the University of Berne Small Animal Radiology Department for an ultrasound examination of the abdomen to confirm a suspected diagnosis of Cushing's syndrome. The patient had bilateral alopecia, was apathic and obese. Ultrasonographically, a tumor of the left adrenal gland, obstruction of the left ureter by an ureterolith, as well as hydronephrosis of the left kidney were detected. During surgery to relieve the ureteral obstruction the adrenal gland tumor was removed. The guinea pig died post-operatively due to blood loss. The left adrenal gland tumor was found histopathologically to be an adenoma and the right adrenal gland also had multiple small adenomas, but grossly appeared normal. The ureterolith was analyzed and found by x-ray diffraction to consist of calcium carbonate.  相似文献   

Developmental changes in pituitary–thyroid axis, and formation of gonads in leptocephali and glass eels of Anguilla spp.     

Yuichi Ozaki  Hiromi Okumura  Yukinori Kazeto  Toshitaka Ikeuchi  Shigeho Ijiri  Masaki Nagae  Shinji Adachi and  Kohei Yamauchi 《Fisheries Science》2000,66(6):1115-1122

SUMMARY: Pituitary, thyroid gland and gonads in leptocephali of Japanese eel Anguilla japonica (19.8–32.6 mm in total length), A. obscura (45.0 mm), and A. bicolor pacifica (49.5 mm) and those in glass eels of the Japanese eel were histologically and immunohistochemically examined in order to observe the developmental changes of these endocrine organs in the Anguillidae. The pituitary, consisting of adenohypophysis and neurohypophysis in Japanese eel leptocephali over 22.5 mm, did not contain thyroid stimulating hormone (TSH) immunoreactive cells. Such cells were, however, detectable in the more developed pituitaries of leptocephali of A. obscura and A. bicolor pacifica and in those of glass eels. Conversely, thyroxine (T₄)-immunoreactive thyroid follicles could be detected in all specimens, both leptocephalic and glass eel. Only in glass eels, gonads were found in the body cavity, and these gonads harbored one or two primordial germ cells (PGC) per cross-section. Our results indicate that thyroid hormones (TH) production started prior to TSH production, and that TSH and TH are both secreted during the metamorphosis from leptocephalus to glass eel. Therefore, it is plausible that the TSH–TH axis is involved in the metamorphosis from leptocephalus to glass eel, but not in the early growth from preleptocephalus to leptocephalus.  相似文献   

荷斯坦乳牛乳腺和乳腺上皮细胞中Toll样受体及辅助因子编码基因的检测     

王亨 孟霞邱昌伟  吴培福韩超 齐长明 《中国兽医科技》2007,37(10):882-886

参照牛TLR4、TLR2、CD14、MD-2基因序列设计了相应基因的引物。采用RT-PCR技术检测了体外培养的荷斯坦乳牛乳腺和乳腺上皮细胞中Toll样受体TLR4、TLR2及辅助因子CD14、MD-2基因。结果显示,乳腺上皮细胞中存在TLR4、TLR2、CD14和MD-2四个基因的表达,而乳腺中除MD-2未检测到外,其余3个基因均扩增成功。说明该受体及辅助因子可能参与了乳腺的先天性免疫防御。该研究为探讨乳腺的先天性免疫及乳腺上皮细胞在乳腺先天性免疫中的作用奠定了基础。  相似文献   

牛蛙和中华鳖甲状腺、肾上腺显微结构的比较研究     

胡毅  钟蕾  葛熹凯 《现代农业科技》2009,(16):267-269

将健康的活体牛蛙、中华鳖的肾上腺、甲状腺取出,用Bouin氏液固定,包埋,切片,H-E染色法染色。比较观察后发现：牛蛙的肾上腺由成堆的细胞小体和管状体构成,中华鳖则由皮质和髓质构成,两者有明显的区别。但甲状腺牛蛙和中华鳖都是由滤泡组成,滤泡周围由丰富的毛细血管和网状纤维构成,其间充满均质状的胶体物质。  相似文献   

幼年牦牛松果体的光镜和电镜观察     

王正波  余四九谭娟 崔燕 《中国兽医科技》2007,37(9):802-806

利用光镜和电镜技术对幼年牦牛松果体的组织形态结构特征进行了研究。结果表明，光镜下，幼年牦牛松果体由松果体细胞、少量的神经胶质细胞、毛细血管和神经等组成。电镜下，松果体细胞的电子致密度低，细胞质内含有丰富的线粒体、粗面内质网、滑面内质网、高尔基复合体、微管、微丝和核糖体，典型异质细胞器突触带呈球形，多位于质膜附近。神经胶质细胞内的线粒体丰富，胞体突起呈球形膨大伸入到松果体细胞之间。松果体细胞以及神经胶质细胞间均存在突触和连接复合体。牦牛松果体内的毛细血管为连续型，远腹侧血管周围可见色素细胞。  相似文献   

生物反应器培养BHK-21悬浮细胞增殖伪狂犬病病毒工艺优化     

王家敏  李自良  马芳芳  康碧静  田玲  李倬  马忠仁  乔自林 《畜牧兽医学报》2022,53(11):3936-3947

旨在筛选伪狂犬病病毒（PRV）敏感的BHK-21细胞并分析其生长和病毒增殖特性,优化反应器中BHK-21悬浮细胞的培养和病毒增殖条件,建立生物反应器培养BHK-21悬浮细胞增殖PRV工艺。本研究利用响应面和单因素优化法,以细胞生长动力学特性、TCID₅₀病毒滴度等参数为指标,优化1.2 L生物反应器中BHK-21悬浮细胞的最佳培养和增殖病毒条件,在5 L生物反应器中进一步批培养验证。结果显示,筛选获得PRV高敏感的BHK-21-02贴壁细胞和BHK-21-XF02悬浮细胞各1株,BHK-21-XF02悬浮细胞在含3%血清的SLM-BHK低血清培养基和SFM-BHK无血清培养基中均能实现良好的生长和病毒增殖。利用响应面法优化得到1.2 L反应器最佳培养条件为接种密度1.20×10⁶cells·mL^-1、搅拌转速120 r·min^-1、DO值40%,5 L反应器批培养72 h细胞密度可达（7.61±0.18）×10⁶ cells·mL^-1、细胞活率为（96.93±1.18）%。利用单因素法优化得到1.2 L反应器最佳病毒增殖条件为MOI 0.001、培养温度37℃、细胞密度2.0×10⁶cells·mL^-1、搅拌转速80 r·min^-1,5 L反应器批培养接毒后48 h病毒滴度达到最大值（7.13±0.11） lgTCID₅₀·mL^-1。本研究可为PRV疫苗相关研究和规模化生产提供参考。  相似文献   

奶牛乳腺上皮细胞的原代培养及其生物学特性分析   总被引：1，自引：0，他引：1  

孙培皓  赵鑫哲  吴涵潇  吕策  杨利国  梁爱心 《畜牧兽医学报》2022,53(2):447-458

旨在从奶牛乳腺组织中分离原代乳腺上皮细胞（bovine mammary epithelial cells,BMECs）并传代培养后探究其生物学特性。本研究从屠宰场采集健康泌乳奶牛乳腺并采用改进的酶消化法从乳腺中分离得到原代奶牛乳腺上皮细胞,通过形态学观察、免疫荧光以及染色体核型分析的方法对其进行鉴定。同时,研究第3、第6和第9代乳腺上皮细胞的生长曲线、群体倍增时间和冻存复苏活力,检测不同代次细胞分泌乳蛋白、乳脂、乳糖的功能及泌乳相关基因的表达。结果表明,所分离的奶牛乳腺上皮细胞纯度较好,细胞生长呈现S型,3个代次细胞的群体倍增时间依次为34.87、41.45和65.04 h,冻存复苏活力为88%~93%;在细胞分泌功能方面,诱导培养2 d后均能检测到酪蛋白、甘油三酯和乳糖,且各代次间无显著差异;此外,3个代次的细胞诱导后均能表达乳成分合成相关基因。本研究成功培养了原代奶牛乳腺上皮细胞,并证明直到第9代细胞仍然具有正常的生物学功能,为体外探究乳腺细胞增殖与分化机制提供了良好的试验材料和技术支撑。  相似文献   

Effect of anaesthesia on cell-mediated immunity in dogs undergoing mastectomy for mammary cancer     

《Veterinary anaesthesia and analgesia》2022,49(3):265-274

ObjectiveTo investigate if anaesthesia for canine cancer mastectomy further influences host cell-mediated immunity (CMI) promoting cancer progression.Study designA randomized, controlled, blinded clinical study.AnimalsA total of 20 bitches with malignant mammary tumours of clinical stage II or III undergoing the same type of mastectomy (regional mastectomy).MethodsDogs were randomly allocated to one of two anaesthetic groups (10 per group). The anaesthetic protocol of group A used minimally immunosuppressive drugs (tramadol, robenacoxib, propofol), whereas that of group B (control) used more immunosuppressive drugs (morphine, fentanyl, thiopental, isoflurane). For each animal, measurements of white blood cells (WBCs), neutrophils and lymphocytes, and flow cytometric assessment of T cells (CD3⁺), helper T cells (CD4⁺), cytotoxic T cells (CD8⁺) and CD5^low+ T cells were performed prior to anaesthesia (day 0) and on days 3 and 10 postsurgery. Data were analysed using a General Linear Model for repeated measures and presented as mean ± standard deviation, p ≤ 0.05.ResultsIn all animals, on day 3, WBCs and neutrophils were significantly increased (p < 0.0005), while flow cytometry revealed significantly decreased relative percentages of T cells (CD3⁺) (p = 0.003) and their subpopulations CD4⁺ (p = 0.006), CD8⁺ (p = 0.029) and CD5^low+ (p = 0.031). Specifically, on day 3, the cytotoxic T cells (CD8⁺) were significantly decreased (p = 0.05) only in group B, whereas the CD4⁺ (p = 0.006) and CD5^low+ (p = 0.008) T cells in group A. The only significant difference between groups was found preoperatively in the CD4⁺/CD8⁺ ratio, which was higher in group A (p = 0.006).Conclusions and clinical relevanceIn dogs with mammary cancer undergoing regional mastectomy, a significant decrease in components of CMI was observed on day 3 postsurgery in both anaesthetic groups. Some indication, however, for better preserved cellular immunity by less immunosuppressive anaesthetic/analgesic drugs was detected, rendering their use advisable.  相似文献