基因编辑动物模型在人类疾病研究中的应用

近年来,随着以CRISPR/Cas9为代表的多种CRISPR系统的开发和不断改进,基因编辑技术逐渐完善,并广泛应用于人类疾病动物模型的制备。基因编辑动物模型为人类疾病的发病机理、病理过程以及预防和治疗等方面的研究提供了重要的素材。目前,用于人类疾病研究的基因编辑动物模型主要有小鼠、大鼠为代表的啮齿类动物模型和以猪为代表的大动物模型。其中啮齿类动物在机体各方面与人类差别较大,且寿命短,无法对人类疾病的研究和治疗提供有效评估和长期追踪;而猪在生理学、解剖学、营养学和遗传学等各方面与人类更接近,是器官移植和人类疾病研究领域重要的动物模型。文中主要介绍了基因编辑动物模型在神经退行性疾病、肥厚心肌病、癌症、免疫缺陷类疾病和代谢性疾病等5种人类疾病研究中的应用情况,以期为人类疾病研究及相关动物模型的制备提供参考。     

子宫内膜异位症动物模型研究进展

为更好的把动物模型应用于子宫内膜异位症的病因、发病机制及治疗等方面的研究,本文将对子宫内膜异位症实验动物的选择、传统动物模型、新型特殊动物模型的构建方法及优缺点进行综述,以期为子宫内膜异位症疾病动物模型的研究提供参考。     

成年生长激素缺乏动物模型与骨代谢相关研究进展

目的探讨成年生长激素缺乏动物模型的几种建立方法,为实验研究和治疗因生长激素缺乏引起的骨代谢异常提供良好的模型。方法通过查阅文献,对成年生长激素缺乏动物模型的建立方法进行综述和评价。结果成年生长激素缺乏动物模型可分为自发性生长激素缺乏动物模型、垂体切除动物模型、基因敲除模型三种。结论垂体切除动物模型价格低廉,可操作性强,但受影响的因素较多,不适合于生长激素与骨代谢之间关系的研究;自发性生长激素缺乏动物模型与基因敲除动物模型价格高昂,但特异性缺乏生长激素,利于研究生长激素缺乏对骨代谢的影响。     

心气虚病证动物模型及其评价体系的构建

目的　研究心气虚病证动物模型及其评价体系的构建。方法　移植心肌梗塞致心力衰竭大鼠的制作模型 ,运用中西医结合虚证和血瘀证的全国统一诊断标准、将其定性的问诊内容代以定量的同等意义的指标测试进行心气虚证动物模型评价。结果　本研究制作的动物模型再现了从一个正常大鼠→以血瘀为主要损伤→形成心气虚证的过程 ;心气虚证动物模型具有时相性、功能性和与功能相关的组织结构物质性改变。结论　将中西医临床和或基础研究中规范的、成熟的、统一的方法和标准引入中医动物模型的研究 ,并注重吸取中医已取得的临床经验和研究成果 ,是指导中医病证动物模型建立的基本思路     

常用实验动物肝癌模型研究进展

肝癌动物模型是研究肝癌发病病因和机理的重要平台和手段,在肝癌研究过程中,肝癌动物模型的建立起着非常重要的作用,建立和提供良好的肝癌动物模型为今后进一步研究肝癌的致病机理,指导临床肝癌的诊断和治疗提供理论参考。     

肺癌动物模型的制备与应用

肺癌动物模型在研究人类肺癌的诊断和治疗等方面发挥着非常重要的作用。根据制备方法及研究目的的不同,肺癌动物模型可分为自发性、诱发性、移植性和转基因动物模型,每种动物模型都有其特征和应用特点。本文旨在介绍肺癌动物模型的制备方法、应用以及近年来的研究进展。     

鼠疫动物模型:经典与问题

一直以来,鼠疫菌的研究多围绕致病机制、疫苗效果评价及鼠疫治疗方案选择展开,而合理有效的鼠疫动物模型正是这些实验研究的基础与前提。研究者构建了包括小鼠、豚鼠及非人灵长类模型在内的多种动物模型,而这些模型也为人类认识、防御及治疗鼠疫菌提供了帮助,特别是一些经典动物模型沿用至今。但随着研究的深入,有些动物模型由于存在某种问题而逐渐被淘汰。本文就鼠疫菌动物模型构建以来,不同模型的优势与不足及其应用展开综述,以期为研究者提供参考。     

缺血性中风动物模型研究现状

利用实验动物或细胞模型完全模拟人类的中风十分困难,动物模型与临床的拟合具有重要意义。本文对目前缺血性中风动物模型研究中的实验动物选择、模型评价标准及造模方法 ,以及主要局灶缺血模型的优缺点进行述评,为缺血性中风的基础和应用研究选择合适的实验动物模型提供参考。     

乳腺癌实验动物模型的制备与应用

乳腺癌实验动物模型在研究人类乳腺癌的生物学行为及治疗等方面起着非常重要的作用。根据制备方法及研究目的的不同,乳腺癌实验动物模型可分为自发性、诱发性、移植性、转基因性及乳腺癌骨转移等动物模型,每种动物模型都有各自的特点和应用条件。     

SARS病毒感染动物模型

SARS-CoV感染动物模型的建立可加深对SARS病原学的了解和发病机制的研究,加速实验室诊断技术的建立、抗病毒药物的筛选和疫苗的开发,同时也有助于给该病一个更精确的定义。可以说SARS动物模型的建立,不但是SARS研究的瓶颈问题,其应用更是贯穿SARS研究的整个过程。到目前为止,已经报道有4种非人灵长类动物(恒河猴、食蟹猴、绒猴、非洲绿猴)和6种啮齿类动物(大鼠、小鼠、豚鼠、田鼠、仓鼠、转基因鼠),以及雪貂、家猫等可以作为SARS动物模型用于实验研究,并已经开始利用动物模型进行疫苗和药物的安全性和有效性评价。本文就已报道的各类SARS动物模型进行综述,并根据动物模型和SARS患者的比对,提出动物模型建立的技术要点。     

ERβ对子宫内膜异位症小鼠模型的影响

目的通过建立子宫内膜异位症小鼠模型探讨雌激素β受体对子宫内膜异位症的影响。方法利用雌激素β基因敲除小鼠建立自体子宫内膜异位模型;应用人不同的组织在SCID小鼠建立子宫内膜异位症模型后,注射雌激素β受体激动剂WAY-200070,观察其对异位病灶生长的影响。结果比较30只雌激素β受体基因敲除小鼠及22只同源未敲除小鼠异位病灶生长及组织细胞形态无明显差异（P〉0.05）;雌激素β受体激动剂WAY-200070对不同类型的SCID小鼠内异症病灶生长影响无明显差异（P〉0.1）。结论雌激素β受体对子宫内膜异位病灶的形成影响作用微弱。     

Mouse model of surgically-induced endometriosis by auto-transplantation of uterine tissue

Endometriosis is a chronic, painful disease whose etiology remains unknown. Furthermore, treatment of endometriosis can require laparoscopic removal of lesions, and/or chronic pharmaceutical management of pain and infertility symptoms. The cost associated with endometriosis has been estimated at 22 billion dollars per year in the United States. To further our understanding of mechanisms underlying this enigmatic disease, animal models have been employed. Primates spontaneously develop endometriosis and therefore primate models most closely resemble the disease in women. Rodent models, however, are more cost effective and readily available. The model that we describe here involves an autologous transfer of uterine tissue to the intestinal mesentery (Figure 1) and was first developed in the rat and later transferred to the mouse. The goal of the autologous rodent model of surgically-induced endometriosis is to mimic the disease in women. We and others have previously shown that the altered gene expression pattern observed in endometriotic lesions from mice or rats mirrors that observed in women with the disease. One advantage of performing the surgery in the mouse is that the abundance of transgenic mouse strains available can aid researchers in determining the role of specific components important in the establishment and growth of endometriosis. An alternative model in which excised human endometrial fragments are introduced to the peritoneum of immunocompromised mice is also widely used but is limited by the lack of a normal immune system which is thought to be important in endometriosis. Importantly, the mouse model of surgically induced endometriosis is a versatile model that has been used to study how the immune system, hormones and environmental factors affect endometriosis as well as the effects of endometriosis on fertility and pain.     

Record review of baboons with histologically confirmed endometriosis in a large established colony

Spontaneous endometriosis was diagnosed in 43 baboons over a 14-year period. Thirty-seven have died; five remain alive; one was sold and lost to follow-up. The average age at diagnosis was 17.2 years; 29 (67%) were between 12 and 21 years of age. Fifteen (35%) were diagnosed by biopsy and received surgical excision of the endometriotic tissue; four of these were identified during caesarian section, confirming one prior report of endometriosis in pregnant animals. Twenty-eight (65%) were diagnosed at or shortly preceding necropsy. When diagnosed by a palpable abdominal mass, there was a significantly greater likelihood the animal died or was killed as a result of complications of endometriosis. When diagnosis was at necropsy, there was a significantly greater likelihood that the animal died from causes unrelated to endometriosis. Early identification with surgical removal appears to provide a benefit for both survival and delivering offspring after diagnosis. In twenty-one baboons (49%), endometriosis affected multiple sites within the peritoneal cavity. In the remaining baboons, lesions were more localized. Ovarian involvement was seen in sixteen (37%) of these baboons. This paper is the first to describe significant ovarian involvement in baboons, previously considered a limitation of the usefulness of this species as an animal model. We also describe the first reported endometriosis seeding of an abdominal surgery scar in a baboon. Many of these baboons were middle aged, had few or no offspring, or had evidence of a long duration of uninterrupted menstrual cycles, consistent with risk factors for women. Endometriosis was an incidental finding in 17 (40%) of these baboons, consistent with previous reports of minimal endometriosis as a common asymptomatic finding in baboons and in women. Overall, endometriosis in baboons presents a spontaneously occurring animal model that shares important features with the disease in women and the rhesus macaque.     

大鼠子宫内膜异位模型的建立与组织学观察

目的为开发诊治子宫内膜异位症(endometriosis,EMT)的新药研究提供理想的动物模型。方法取雌性未交配性成熟大鼠,术前雌激素诱导,麻醉开腹取部份右侧子宫,将内膜种植于左腹壁内,术后16周取出包块,进行组织形态学、组织化学观察。结果异位内膜在腹壁内生长,呈隆起囊状小包块,内有黏液,具有正常子宫内膜基本组织结构,囊腔较大。异位内膜中有糖原、RNA的存在。结论该手术方法建立的子宫异位内膜生长良好,术后一周就可摸及包块大小,为开发研究子宫内膜异位症的新药提供了方便。     

Pharmacologic properties of CDB(VA)-2914

CDB(VA)-2914 (17alpha-acetoxy-11beta-(4-N,N-dimethylaminophenyl)-19-norpregna-4,9-diene-3,20-dione) is a synthetic steroid that demonstrates potent progesterone antagonist activity in vitro and in vivo. Its binding and antagonist potency with respect to the glucocorticoid receptor is significantly reduced compared to that of mifepristone, indicating that CDB(VA)-2914 belongs to a new class of dissociated progesterone receptor modulators that have reduced antiglucorticoid activity. The pharmacological effects of CDB(VA)-2914 have been examined in a variety of animal models, the results of which are reviewed in this paper. CDB(VA)-2914 inhibits ovulation in rats in a dose-dependent manner upon single-dose oral administration and exhibits antifertility activity during continuous low-dose administration. CDB(VA)-2914 is also effective in animal models of postcoital contraception. This paper also presents the results of metabolism studies undertaken to link the results of the animal models to potential human applications. Because of its unique pharmacological profile, CDB(VA)-2914 is a promising candidate for use in contraception as well as treatment of uterine fibroids and endometriosis.     

雌激素受体β基因敲除小鼠子宫内膜异位症模型的建立

目的建立雌激素受体β基因敲除小鼠子宫内膜异位症模型,为进一步研究雌激素受体β基因在子宫内膜异位症发生发展过程中的作用提供平台。方法用外科手术方法对16只β基因敲除小鼠进行自体子宫移植,术后14d、21d取病灶组织进行光镜观察分析。结果建模成功率达95．8％,可形成明显囊肿,内含囊液,囊肿内壁有子宫内膜上皮细胞生长。结论应用本方法可建立稳定的子宫内膜异位症转基因小鼠模型,便于研究雌激素受体β亚型及其相关基因、蛋白在子宫内膜异位症发生发展过程中的作用机制。     

血清蛋白质指纹图谱模型在子宫内膜异位症诊断中的应用

用表面加强激光解析电离飞行时间质谱(SELDI-TOF-MS)和蛋白质芯片检测子宫内膜异位症(endometriosis,EM)患者血清蛋白质指纹图谱,探讨诊断模型在EM诊断中的临床应用价值。用SELDI-TOF-MS技术和H4蛋白质芯片检测16例EM和16例正常女性的血清蛋白质指纹图谱,并建立诊断模型。然后,对16名健康人和16例EM患者样本进行盲法测试验证该模型。筛选出4个有明显表达差异的蛋白质,其质荷比(m/z)分别为8141、6096、5894、3269。建立的诊断模型对EM检测的灵敏度为87.5%(14/16),特异性为93.75%(15/16),总准确率为90.625%(29/32)。SELDI-TOF-MS对小样本的EM诊断具有较高的敏感性和特异性,在EM的诊断及标志物筛选等方面具有较好的诊断价值。     

子宫内膜异位症大鼠模型的建立及其病理学改变

目的构建子宫内膜异位症（内异症）大鼠动物模型,为阐明内异位症发病机理以及寻找有效的治疗方法提供理想的动物模型。方法取性成熟雌性Sprague-Dawley（SD）大鼠30只,通过手术将大鼠自体子宫组织移植到子宫旁韧带上,建立诱发型内异症大鼠动物模型。术后8周,再次剖腹观察异位组织的存活情况、病灶大小、与周围组织的粘连程度以及病理学变化。结果25只大鼠有明显的异位病灶。所有病灶都与周围组织有不同程度的粘连,病灶外观呈囊泡状。光镜观察见大部分异位子宫内膜形态和结构与在位子宫内膜基本相同,但内膜细胞、间质细胞、腺体,与在位内膜相比较少。少数病灶只有上皮组织或只有问质组织。结论自体子宫移植法可成功建立内异症大鼠模型。     

Synthesis and biological evaluation of water-soluble progesterone-conjugated probes for magnetic resonance imaging of hormone related cancers

Progesterone receptor (PR) is strongly associated with disease prognosis and therapeutic efficacy in hormone-related diseases such as endometriosis and breast, ovarian, and uterine cancers. Receptor status is currently determined by immunohistochemistry assays. However, noninvasive PR imaging agents could improve disease detection and help elucidate pathological molecular pathways, leading to new therapies and animal disease models. A series of water-soluble PR-targeted magnetic resonance imaging (MRI) probes were synthesized using Cu(I)-catalyzed click chemistry and evaluated in vitro and in vivo. These agents demonstrated activation of PR in vitro and preferential accumulation in PR(+) compared to PR(-) human breast cancer cells with low toxicity. In xenograft tumor models, the agents demonstrated enhanced signal intensity in PR(+) tumors compared to PR(-) tumors. The results suggest that these agents may be promising MRI probes for PR(+) diseases.