Preparation of monoclonal antibodies against mouse alpha-fetoprotein and preliminary study on their application

原发性肝癌是我国高发的恶性肿瘤之一，每年新发和死亡患者均占全球总数的一半以上。根据全国肿瘤登记中心最新公布的2015年《肿瘤登记年报》数据，我国肝癌新增病例为46.61万，占所有恶性肿瘤的第4位（10.86%），而死亡病例为42.21万，占所有癌症死亡的第3位（15.00%），仅次于肺癌和胃癌。尽管近年来肝癌的诊断和治疗有了一定的进展，其预后仍然不理想，五年生存率极低。究其原因，大部分肝癌患者往往在疾病的晚期才被诊断出来，有些甚至发生了远端转移，因而错过了最佳治疗时机；此外，目前几乎没有针对中晚期肝癌有效的治疗手段。在这种情况下，迫切需要开发肝癌早期诊断试剂及新型靶向药物，提高治疗效率。 甲...Liver cancer is one of the most common cancers, more than half new cases and deaths occur in China. According the data from National cancer registry in 2015, liver cancer is the fourth most common cancer with high incidence rate (466.1 per 100,000), and is the third leading cause of cancer death (422.1 per 100,000). Although great advances have been made in diagnosis and treatment of liver cancer ...学位：理学硕士院系专业：生命科学学院_水生生物学学号：2162013115248

赤点石斑鱼促甲状腺激素TSHβ启动子的克隆及其在斑马鱼胚胎原始性腺和垂体中定位表达

促甲状腺激素(Thyroid-stimulating hormone,TSH)具有调节生长,发育,代谢和生殖的功能,但是在低等脊椎动物中的功能研究较少。斜带石斑鱼(Epinephelus coioides)促甲状腺激素TSHβ除在垂体中表达外还在性腺中表达,进一步发现TSHβ也在赤点石斑鱼(Epinephelus akaara)性腺中表达。且在人工诱导的赤点石斑鱼中,随着雄性化的转变,TSHβ转录水平升高。为研究石斑鱼TSHβ特殊表达方式的调控机制,采用基因组步移技术扩增获得赤点石斑鱼TSHβ5′端序列

Survivinの甲状腺濾胞上皮細胞増殖および分化に対する影響

　Survivin is one of the IAP (inhibitor of apoptosis proteins), which is expressed during embryonic development, but is not expressed in terminal differentiated adult tissue. Overexpression of survivin is observed in a variety of human tumors including thyroid cancers. We examined the role of survivin on the proliferation and differentiation of thyroid epithelial cells. Rat thyroid epithelial cell line FRTL-5 was stably transfected with human sense and antisense survivin genes. FRTL-5 cells transfected with a survivin sense gene (S cells) proliferated without TSH. In the presence of TSH, the proliferation of S cells was significantly faster than that of cells transfected with an empty vector (E cells) or cells transfected with a survivin antisense gene (AS cells). The proliferation of AS cells were retarded as compared with that of E cells. Western blot analysis demonstrated that phosphorylation of Rb protein at serine 795 was enhanced in S cells. Activation of E2F-1 was also enhanced in S cells as demonstrated by gel shift assay. Northern blot analysis showed that expression of Pax-8 mRNA was significantly enhanced in S cells, and was significantly decreased in AS cells. Expression of thyroglobulin (Tg) and TSH receptor (TSHR) mRNA was decreased in AS cells. There was no significant change in TTF-1 and TTF-2 mRNA expression. These observations suggest that survivin enhances the proliferation of thyroid epithelial cells through activation of E2F-1, and also enhance the expression of Pax-8, which then affect the expression of thyroid specific proteins. Survivin appears to play an important role in the proliferation as well as differentiation of thyroid epithelial cells

Hira基因产物在银鲫和彩鲫卵子发生过程中的动态变化

为进一步研究Hira基因在卵子发生和雌核发育过程中的作用,通过原位杂交和免疫荧光定位的方法检测了Hira mRNA和蛋白质在雌核发育银鲫和两性生殖彩鲫卵子发生过程中的动态变化。结果表明,银鲫和彩鲫卵子发生过程中Hira基因转录产物的变化基本一致,在Ⅰ期卵母细胞的细胞核中大量表达,至Ⅱ期卵母细胞时转至细胞质中均匀分布,在Ⅲ期卵母细胞中,杂交信号逐渐移向细胞的周边,到Ⅳ期时随着卵黄物质大量积累,杂交信号几乎不见。HIRA蛋白在银鲫和彩鲫卵子发生过程中的变化略有差别。HIRA蛋白在银鲫Ⅰ期卵母细胞中没有表达,

Toxicity of BaP exposure during embryonic stage in zebrafish: the mechanisms involved

多环芳烃（PolycyclicAromaticHydrocarbon，PAH）是环境中一类分布广泛的持久性有机污染物。苯并（a）芘（Benzo(a)pyrene，BaP）作为代表性的PAH具有包括神经毒性和生殖毒性在内的多种毒性。截至目前，关于BaP神经毒性的研究大多集中于高浓度或短期的暴露实验，而环境水平浓度下、长期的BaP暴露是否能引起同样的神经毒性则研究得较少；此外，胚胎时期低浓度BaP暴露对成年阶段的神经毒性、生殖毒性效应及其机制研究更为少见。因此本研究以斑马鱼为模式动物，分别进行环境水平浓度(0，0.05，0.5，5，50nmol/L)BaP长期暴露和胚胎时期短暂暴露（后正常清水中养...Benzo(a)pyrene (BaP) is a kind of polycyclic aromatic hydrocarbons which are most abundant and ubiquitous persistent organic pollutants (POPs) in environment. So far, most studies of the neurotoxicity concerning BaP focus on high dose exposure or short-term exposure. The effects of neurotoxicity on long-term environmental relevant levels exposure of BaP are still limited. Moreover, little is known...学位：理学博士院系专业：生命科学学院_动物学学号：2162013015409

内分泌扰乱化学品对动物的影响和作用机制

介绍了内分泌扰乱化学品 (EDCs)的作用机制及其对野生动物影响的研究进展。内分泌化学品可能与性激素受体、甲状腺素受体和其他受体 (如芳烃受体 )结合 ,也可能通过影响动物体内激素的代谢而对动物的生长发育起调控作用。各实验室的重复实验不支持环境雌激素之间存在协同作用的结论。已报道过的人体和野生动物生殖缺陷中具有充分证据的暴露 -效应关系的实例为数不多。尽管野生鱼类变性是英国河流中的一个普遍现象 ,但进一步的实验证明污水处理场出水中的天然和合成雌激素足以引起鱼类的变性反应 ,而不是先前怀疑的非离子表面活性

The discovery of new biomarkers of hepatocellular carcinoma

肝癌，尤其是肝细胞型肝癌是一种常见的恶性肿瘤，发现晚且死亡率高，因此及时发现、尽早治疗具有重要的临床意义。而生物标志物可作为早期临床诊断的工具，可显著提高患者的生存率。当前临床常用的肝细胞型肝癌的血清早期诊断标志物主要有甲胎蛋白（AFP）和甲胎蛋白异质体（AFP-L3）等，但这些检测指标的灵敏度和特异性均有待提高。而外泌体是一种与肿瘤的发生、发展密切相关的分泌型囊泡。已有报道显示，许多肿瘤中均可分泌包含特异的、可做潜在生物标志物的外泌体，因此，利用外泌体寻找生物标志物是目前研究的热点。 本课题从肿瘤细胞分泌的外泌体（exosome）的蛋白质组学入手，期望通过临床样本的筛查，找到在肝癌病人血清...Hepatocellular carcinoma (HCC), which has been often in late stage when diagnosed, is a common malignant tumor with a high mortality rate. Early detection and treatment of HCC could significantly increasing survival rate, so it's very important to develop clinical biomarkers for early clinical diagnosis. Nowadays, wildly used clinical early diagnostic biomarkers for HCC are mainly alpha fetoprotei...学位：医学硕士院系专业：药学院_药理学学号：3232014115343

Relationship between Vitamin and Fetal Development

胎儿在母体宫内发育经过受精卵、胚胎、胎儿三个阶段,任何影响母胎健康的病因都影响宫内受精卵、胚胎和胎儿的发育,严重者甚至导致胚胎停育、胎死宫内。缺乏维生素容易诱发人体各种疾病,孕期母体维生素的需要量增加,如果孕期母体维生素摄入不足、补充不够、未得到及时增加或增加过量,都会影响胎儿器官的正常发育,导致胎儿发育不良。有些严重的胎儿畸形是致死性的,因此孕期要注意筛查及时处理。孕期加强母胎保健,重视优生优育,改善孕妇和儿童营养状况,合理调配膳食,适当补充维生素,了解各种维生素的功效和胎儿发育的相关性,有助于降低新生儿出生缺陷、防治妇女儿童营养缺乏病。aFertilized egg, embryo and fetus are three stages of fetus growth in maternal intrauterine. Any factors caused by the mother or the fetus can affect intrauterine growth of fertilized egg, embryo and fetus. Some serious factors could lead to embryonic growth cease, even fetal death. Lack of vitamin can induce human related diseases, and vitamin requirements are increased in pregnant women. If insufficient of vitamin intake in those pregnant women, fetus could suffer from dysplasia. Some severe fetal abnormalities can cause fetal death. So, it is necessary that pregnant women should have fetal developmental screening, and that those pregnant women with vitamin insufficient should be treated in time. The reasonable supplement of vitamins, and deployment of diet, is benefit for them to reduce neonatal defects, and to prevent the nutrition-related diseases in pregnant women

Progesterone regulating glutathione S-transferase Omega-1 expression in the mouse uterine luminal and glandular epithelium during preimplantation period

目的研究谷胱甘肽S-转移酶Omega-1(Gsto 1)在小鼠胚胎着床过程中的表达和孕酮的调节。方法105只CD1小鼠,分为正常妊娠模型和类固醇激素处理模型。正常妊娠模型中,收集妊娠第1~第5天子宫,采用Real-time PCR、原位杂交和Western blotting 3种方法检测Gsto1的表达变化;类固醇激素处理模型均采用卵巢切除2周后的小鼠,又分为雌孕激素处理组、孕酮处理不同时间组和孕酮受体拮抗剂Ru486处理组,所有组中的对照均用芝麻油处理。雌孕激素处理组中,收集芝麻油、雌激素、孕酮、雌激素加孕酮分别处理12h后的子宫;孕酮处理不同时间组中,收集芝麻油和孕酮分别处理1、3、12、24 h后的子宫;Ru486处理组中,收集芝麻油、Ru486、孕酮、Ru486加孕酮分别处理12 h后的子宫。类固醇激素处理模型使用Real-time PCR和Western blotting两种方法检测Gsto1的表达变化。结果 Gsto1主要在妊娠第1~4天的子宫腔上皮及腺上皮中表达,其中,妊娠第1~3天表达量较高,第4天表达量较低,第5天着床点和非着床点均不表达。孕酮诱导Gsto1的表达,雌激素不能诱导Gsto1的表达,并能抑制孕酮对Gsto1的诱导。Ru486降低孕酮对Gsto1的诱导,孕酮处理1、3、12 h均促进Gsto1的表达,但作用24 h后,抑制Gsto1的表达。结论 Gsto1在小鼠妊娠早期子宫腔上皮及腺上皮中表达,雌激素能够拮抗孕酮对Gsto1的诱导,孕酮可以通过孕酮受体调节Gsto1的表达,并且具有短时调节作用。Objective To investigate the expression and regulation of glutathione S-transferase Omega-1( Gsto1) in the mouse uterus during embryo implantation. Methods A total of 105 CD1 mice were divided into the normal pregnancy model and steriod hormone treatment model. Uterus were collected from days 1 to 5 of pregnancy in normal pregnancy model,and Gsto1 expression was detected by Real-time PCR,in situ hybridization and Western blotting. Ovariectomized mice were used in the steriod hormone model after 2 weeks,and divided into estrogen and progesterone treatment group,progesterone treatment course group,progesterone receptor antagonist Ru486 treatment group. Sesame oil was used for the control of all groups. In the estrogen and progesterone treatment group,uterus was collected after sesame oil,estrogen,progesterone,estrogen plus progesterone treatment 12 hours,respectively. In the progesterone treatment course group,uterus was collected after progesterone treatment 1 hours,3 hours,12 hours and 24 hours,respectively. In Ru486 treatment group,uterus was collected after sesame oil,Ru486,progesterone,Ru486 plus progesterone treatment 12 hours. Gsto1 expression was detected by Real-time PCR,and Western blotting in the steriod hormone model. Results Gsto1 was mainly expressed in the uterine luminal and glandular epithelium on days 1 to 4 of pregnancy. Gsto1 expression was high on day 1 to 3,but became lower onday 4. On day 5,Gsto1 expression was not detected at implantation sites and non-implantation sites. Progesterone induced Gsto1 expression. Estrogen did not induce Gsto1 expression,but inhibited the induction of progesterone on Gsto1. Ru486 reduced the induction of progesterone on Gsto1. Progesterone treatment for 1 hour,3 hours,12 hours promoted Gsto1 expression,but after 24 hours,inhibited Gsto1 expression. Conclusion This study suggests that Gsto1 is mainly expressed in the uterine luminal and glandular epithelium during preimplantation. Estrogen inhibits the induction of progesterone on Gsto1.Progesterone enhances Gsto1 expression by progesterone receptor in short time.国家自然科学基金(31401953);; 安徽省教育厅自然科学重点项目(2014A143

A Study on Roles of Slit2 in Hepatocellular Carcinoma Cells

Slit家族蛋白是一种进化上高度保守的细胞外分泌糖蛋白。对Slit的研究最初的重点是它在神经生长的作用，Slit作为配体通过与跨膜受体Roundabout(Robo)结合在中枢神经系统轴突导向中起到重要的作用。近些年的研究发现Slits与Robos在一些肿瘤中异常表达，说明Slit/Robo信号在肿瘤的发生发展过程中起到重要的作用。 Oncomine数据库的检索结果显示，与正常组织相比，肝细胞癌中受体Robo1的表达量有显著的增加，而Slit2的表达量则有一定的差别，多数情况下表现为下调。并且多数研究发现，在不同的肿瘤中配体Slits与受体Robos的表达存在着不一致性，而且Slit/Rob...Slits are a family of secreted glycoprotein conserved in evolution. Early research of Slits was focused on their effects in neuronal guidance. Binding with a transmembrane receptor family of Roundabout (Robo), the ligand Slit plays a critical role in the central nervous system (CNS) axonal guidance. Recent studies showed that both Slits and Robos were abnormally expressed in some cancers, indicati...学位：理学硕士院系专业：生命科学学院_细胞生物学学号：2162012115245