Effects of Photoperiod on Stem Cells Activation in Cashmere Goat Hair Follicles

In order to extend the anagen of cashmere goat hair follicles and increase the production of cashmere,this study was performed with artificially shorten the daylight time among Arbas White cashmere goats. Skin tissue sections from cashmere goats were collected to compare the morphologic changes between artificial daylight and natural daylight,and immunohistochemical method was used to study the hair follicle cell proliferation and important protein expression in related signaling pathways. The results showed that strong cell proliferation occurred in cashmere goat hair follicle cells during artificial daylight,plenty of cytokeratin 15 (K15) positive signals were distributed in the outer root sheath,β-catenin protein was actively expressed in hair matrix and root sheath, indicating that the hair follicles were in the anagen growth phase;Meanwhile,cashmere goat hair follicles under natural daylight were in telogen with weak signals. Above all prove that short photoperiod played an important role in promoting hair follicle growth,the artificial short photoperiod could change hair follicle growth cycle and make hair follicles earlier enter to the anagen growth phase,causing a variety of typical gene expressions during hair follicle growth.     

黄鳍鲷肝细胞体外培养研究

以黄鳍鲷肝脏细胞为材料 ,研究其肝细胞体外培养适宜的 p H、血清浓度、谷氨酰胺等条件 ,并进行传代培养试验。结果表明 ,培养液为 :MEM含 2 0 %小牛血清、15 g/ L L-谷氨酰胺及 p H 6 .9～ 7.1时 ,在 2 3℃中静置培养 ,黄鳍鲷肝细胞生长良好 ,3～ 5 d长满单层 ,细胞主要为成纤维细胞状 ,并能顺利传代。细胞动力学试验表明 :传代细胞 1～ 2代为适应期 ,3～ 5代为旺盛期 ,6代之后表现出衰老现象     

奶牛乳腺上皮细胞三维培养的形态观察

利用原代培养后纯化的奶牛乳腺上皮细胞,以Matrigel为基质胶原,进行奶牛乳腺上皮细胞的三维培养,细胞接种浓度分别为1×104、1×105和1×106个细胞/mL。经过16d培养后,用DAPI对细胞染色,利用荧光显微镜观察细胞生长状况。结果表明,当细胞接种浓度为1×104～1×105个细胞/mL时,细胞形成了团块状结构,出现了类似腔状的腺泡形态。     

家兔黏膜免疫器官肥大细胞在巴氏杆菌感染中的特征——组织化学与电镜观察

对5例巴氏杆菌感染家兔的空肠与圆小囊肥大细胞（mast cell，MC）进行了组织化学与电镜的观察，发现在细菌感染导致的病变组织边缘、病变组织内血管附近结缔组织中及变性细胞周围组织的上皮下与黏膜下层都出现了大量甲苯胺蓝（Toluidine Blue，TB）阳性肥大细胞。电镜下，MC有的直接与变性细胞密切接触，有的则和血管内皮紧密相触，在此部位的血管内可见有淋巴细胞贴附填充。MC胞浆内充盈大量的特征性颗粒，有时这些颗粒向细胞表面突出形成边缘空隙，即形成所谓的脱颗粒管道，将颗粒内容物逐渐排除细胞外，遗留空腔。     

Italian University System has to Match Educational Needs in Adherence to European Standards and Professional Scenario

The effect of a water-soluble fraction (WSF) of a non-pathogenic strain of Mycobacterium phlei was studied in bovine subclinical mastitis (SCM) by measuring the myeloperoxidase and acid phosphatase enzyme levels in the milk leukocytes. Forty-five cows were divided into three equal groups. Group I, consisting of 15 healthy cows, served as the control, whereas groups II and III each contained 15 cows with subclinical mastitis on the basis of a positive reaction in the California mastitis test (CMT). The cows in group II received 100 microg of WSF in 5 ml sterile phosphate-buffered saline, pH 7.4 (PBS) once only, while those in group III received 5 ml sterile PBS daily for 7 days, both treatments being given by the intramammary route. Observations were made up to 30 days after treatment (AT). The CMT of the healthy milk was negative (0), whereas it ranged between 1 and 2 points in SCM. The somatic cell count (SCC) increased significantly (p < 0.05) on day 3, then fell steeply from day 7 up to day 30 AT in the cows in group II. A steady decrease in the total bacterial count (TBC) was observed in the group treated with WSF but the bacterial counts remained high in the groups treated with PBS. The mean acid phosphatase level was enhanced by 119% on day 3 AT in group II but only by 18.7% in the cows in group III. The mean myeloperoxidase level was enhanced by 100% in the cows in group II but only by 18% in those in group III on day 3 AT. This significant reduction in the bacterial load in infected cows caused by intramammary infusion of WSF may be due to activation of the microbicidal activity of the neutrophils, but this requires confirmation.     

鸡胚及禽类细胞系在生物医药领域的应用及研究进展

鸡胚因发育过程清楚,长久以来作为基础和应用科学研究领域重要的实验模型,尤其在鸡胚发育早期绒毛尿囊膜阶段,因其血管丰富,是天然的免疫缺陷宿主,是病理学、药理学和肿瘤学等研究领域的理想实验模型。作者简述了发育早期的鸡胚组织结构,并介绍了鸡胚绒毛尿囊膜在肿瘤研究、血管生成、器官移植、烧伤等疾病机理研究中的应用,以及在鸡胚病理模型基础上进行的抗肿瘤药物筛选的应用。重点介绍了鸡胚及禽类细胞系在病毒繁殖和疫苗生产、治疗性蛋白和单抗生产方面的应用研究进展。多种人源病毒、禽源病毒、支原体等可在鸡胚及禽类细胞上增殖,并用于疫苗生产。作者对常用的禽类纤维原细胞和多能干细胞的发展和特点进行了阐述,并总结了商业化的禽类细胞系来源以及部分易感病毒。鸡胚表达系统能够在目的蛋白特定位点产生人源化糖基,减少目的蛋白对人的过敏反应,且禽蛋廉价易得,可作为生产人用单克隆抗体和治疗性蛋白的合适供体。作者介绍了鸡胚及禽类细胞系在生物医药领域应用的最新进展,并对鸡胚作为动物模型在未来的应用进行了展望。     

NPFFR1基因对鹅卵泡颗粒细胞激素分泌和细胞凋亡的影响研究

神经相关肽受体（RFamide-related peptide receptor,NPFFR1）是促性腺激素抑制激素的主要亲和受体,它在调控动物繁殖方面起着重要作用。为了解NPFFR1对鹅卵巢卵泡发育的作用,本研究以42周龄健康产蛋四川白鹅为试验材料（n=9）,利用RT-qPCR法检测NPFFR1基因在等级前和等级卵泡颗粒细胞中的mRNA表达规律;在颗粒细胞中过表达NPFFR1基因,酶联免疫吸附法检测颗粒细胞上清液（n=9）中雌二醇（estradiol,E2）、孕酮（progesterone,P4）和抗缪勒管激素（anti-Mullerian hormone,AMH）的浓度变化,剩余贴壁细胞作一步法TUNEL检测细胞凋亡情况;转录组测序方法筛选大黄卵泡（8~10 mm）颗粒细胞过表达NPFFR1前后表达差异显著基因,并对差异表达基因进行功能聚类分析。结果显示,除F1等级外,其余等级卵泡颗粒细胞NPFFR1表达量均极显著高于等级前卵泡（P<0.01）;过表达NPFFR1后,等级颗粒细胞上清液中的E2和等级前颗粒细胞上清液AMH的含量显著（P<0.05）降低,但孕酮P4含量变化不显著（P>0.05）;转录组测序共筛选到267个差异表达基因（119个下调,148个上调）,这些基因主要富集在生物节律过程、繁殖进程等生物学过程中;同时,与对照组相比,差异基因AMH显著下调表达（P<0.05）,Clock（clock circadian regulator）、FOS（proto-oncogene,AP-1 trans-cription factor subunit）、Per（period circadian regulator）和ANTXR2（cell adhesion molecule 2）分别极显著（P<0.01）或显著（P<0.05）上调表达。上述试验结果提示,NPFFR1可从激素、细胞凋亡和生物节律等多个环节影响卵泡颗粒细胞,参与调控卵泡的时序等级发育。