人源化小鼠人源细胞检测方法

目的人源化小鼠在人类疾病与再生医学研究中具有广泛应用,为获得一种理想的人源化小鼠检测方法,研究人特异性线粒体序列扩增作为人源化动物模型中人类DNA检测方法的可行性。方法设计人线粒体DNA特异性引物,并用该引物对人脐血造血干细胞嵌合体小鼠进行了检测。结果在人源化动物模型的人类DNA检测中,这种人线粒体DNA的PCR检测方法能够达到理想的特异性与灵敏性,是一种理想的嵌合体小鼠检测方法。结论获得了一种理想的人源化小鼠人源细胞检测方法。     

罗汉果和熟地增加小鼠造血干细胞的数量和功能

目的：罗汉果和熟地具有增强机体免疫力,延年益寿的作用。本文研究罗汉果和熟地对造血干细胞的影响。方法各组小鼠连续喂养罗汉果或熟地3个月,流式细胞仪测量小鼠的外周血、脾脏和骨髓中免疫细胞和造血干细胞的变化;小鼠用半致死剂量的放射线照射后,连续喂养罗汉果或熟地1个月后,流式细胞仪测量罗汉果或熟地对于免疫细胞和造血干细胞损伤修复的作用。结果分析检测结果发现,与对照组相比,长期喂养罗汉果或熟地的小鼠骨髓和外周血中B细胞的比例降低,粒细胞的比例增加,T细胞没有明显变化;骨髓中造血干细胞的数量增加,尤其是长期造血干细胞数量增加显著。半致死剂量的放射线照射后,罗汉果或熟地喂养1个月,小鼠的粒细胞和T细胞都有明显的改善,造血干细胞的数量明显增加。结论罗汉果或熟地长期服用会增加小鼠造血干细胞的数量和功能,促进了辐射所致的骨髓抑制小鼠的造血干细胞的恢复。     

IL-33转基因小鼠骨髓造血干／祖细胞向髓系细胞分化的能力增强

目的：利用IL-33转基因小鼠研究IL-33对造血干/祖细胞的增殖和分化影响。方法利用流式细胞仪分析IL-33转基因小鼠及同窝野生对照小鼠的外周血、脾脏、骨髓细胞的免疫表型及造血干细胞分化不同阶段细胞的数量变化;利用体外成克隆实验和细胞周期分析研究IL-33对于造血干细胞增殖能力的影响。结果与野生型小鼠相比,IL-33转基因小鼠B细胞和T细胞在外周血中都明显降低,粒细胞在外周血和骨髓中都有明显增加;IL-33转基因小鼠的骨髓造血干细胞和多能祖细胞数量减少,共同淋系祖细胞数量减少,共同髓系祖细胞和粒单系祖细胞数量增加;IL-33转基因小鼠的造血干细胞处于S-G2-M的细胞增多;体外单克隆实验发现IL-33转基因小鼠造血干细胞形成的集落数增加。结论 IL-33转基因小鼠造血干细胞增殖能力增强,更易向髓系细胞分化。     

贞芪扶正胶囊对慢性苯中毒小鼠骨髓细胞的影响

为了观察贞芪扶正胶囊对慢性苯中毒(CBP)小鼠中骨髓细胞有丝分裂指数、骨髓造血干细胞表型Sca-1和CD34~+的影响,并探讨其造血调控作用的可能机制。本试验将70只8~12周龄健康雄性昆明种小鼠,按随机数字表法分为5组:对照组、模型组、贞芪扶正组、西药组、贞芪扶正联合西药组,每组14只(实际有效数量每组10只)。将小鼠置于人工模拟高苯浓度(苯浓度=2 000 mg/m~3)环境,复制慢性苯中毒模型。采用用全自动血细胞分析仪、Giemsa染色法、流式细胞仪分别检测贞芪扶正胶囊对苯中毒小鼠外周血、骨髓细胞有丝分裂指数、造血干细胞表型Sca-1和CD34~+的影响。结果表明贞芪扶正联合西药组能提高白细胞等含量;激活小鼠骨髓造血干细胞有丝分裂指数,使Sca-l和CD34~+抗原的表达升高,形成更为原始的造血干细胞,从而造血干细胞数量增加(p0.05),改善慢性苯中毒小鼠的外周血象。本研究表明贞芪扶正胶囊和西药联合使用效果明显,促进抗原表达明显改善中毒现象,安全有效,值得临床应用推广。     

Exo-1基因对端粒酶缺陷小鼠造血干细胞植入效率的影响

目的利用不同的造血干细胞移植方式,探讨Exo-1基因缺失对端粒酶基因敲除小鼠的造血干细胞植入效率的影响。方法以CD45.1小鼠的骨髓细胞或骨髓造血干细胞为供体,以端粒酶基因敲除小鼠或Exo-1基因和端粒酶基因双敲除小鼠为受体,在给予不同剂量X线照射或不照射的情况下,重复进行静脉注射全骨髓细胞或分选的骨髓造血干细胞(c-kit+、Sca-1+、lineage-,KSL),于移植后1个月取外周血,流式分析嵌合率。结果未经X线照射及1 Gy、2 Gy照射情况下,端粒酶基因缺陷受体小鼠的外周血中供体来源的细胞嵌合率较低;6 Gy照射后,供体来源的外周血细胞嵌合率仍低于50%,而且端粒酶基因缺陷受体小鼠在移植后1个月内死亡较多;3 Gy照射可形成较高嵌合率,Exo-1基因缺失对端粒酶缺陷小鼠的造血干细胞植入效率的影响不显著。结论以端粒酶缺陷小鼠作为衰老模型研究造血干细胞植入效率时,3 Gy X线照射能够有效地形成较高的外周血供体细胞的嵌合率,但是Exo-1基因没有进一步提高造血干细胞在端粒酶敲除小鼠的植入效率。     

Cramp基因敲除对小鼠骨髓造血干细胞的影响

目的研究Cramp基因敲除在衰老过程中对小鼠造血干细胞的作用。方法应用流式细胞仪分析3月龄及12月龄Cramp基因敲除小鼠及同窝野生型小鼠的骨髓造血干细胞的比例及不同发育阶段B淋巴细胞的比例。结果与野生型小鼠相比,12月龄Cramp基因敲除小鼠的骨髓长期造血干细胞增多,多潜能造血祖细胞减少;前体B淋巴细胞和未成熟B淋巴细胞减少,成熟B淋巴细胞增多。结论在衰老过程中,Cramp基因敲除对骨髓造血干细胞及B淋巴细胞发育有重要影响。     

Cramp过表达对小鼠骨髓造血干细胞功能的影响

目的研究Cramp蛋白过表达对小鼠骨髓造血干细胞自我更新和分化能力的影响。方法应用流式细胞仪分析Cramp过表达转基因小鼠及同龄野生型小鼠的骨髓、脾脏、胸腺等组织器官中各种细胞的比例;分选骨髓造血干细胞,体外培养,观察其克隆形成能力。结果与野生型小鼠相比,Cramp过表达转基因小鼠的骨髓、脾脏、胸腺等组织器官中各种细胞的比例、骨髓造血干细胞的克隆形成能力等均无明显变化。结论本研究中,Cramp过表达转基因小鼠骨髓造血干细胞的分化能力、克隆形成能力无明显变化。     

人胎肝细胞裂解液对照射小鼠多能造血干细胞的影响

本文采用内、外源脾结节形成测定法和流式细胞术观察了人胎肝细胞裂解液FLL及其峰组分在γ射线照射小鼠多能造血干细胞损伤修复中的作用。证明照后24h给予FLL及峰组分4—5能明显增加受照小鼠残存造血干细胞形成CFU-S的能力,促进处于G1/0期的造血干细胞提前进入增殖状态。     

Gadd45a基因对小鼠造血干细胞功能的影响

目的 Gadd45a基因对小鼠造血干细胞功能的影响。方法流式细胞仪分选小鼠骨髓造血干细胞、体外单克隆培养,竞争性骨髓移植,放射线照射观察生存曲线。结果 Gadd45a基因缺失的小鼠造血干细胞克隆形成能力增强,短期造血重建能力无差异,8.5Gy放射线照射后生存情况无差异。结论 Gadd45a基因对小鼠造血干细胞功能起重要作用。     

利用人脐带血干细胞制备人鼠肝组织嵌合体模型

目的利用人脐带血干细胞研究制备HBV感染人鼠嵌合小鼠模型。方法将人脐血干细胞,经尾静脉分两次注射到裸鼠体内。分别于第7,14,21天取肝组织,进行AFP免疫组织化学检测,分析人肝细胞在裸鼠体内嵌合生长情况,并进行乙肝病毒感染实验,定量检测感染后小鼠血清中AFP、ALB。结果实验组小鼠在第7、14、21天肝脏内AFP持续阳性表达,AFP表达于细胞质内。HBV感染后小鼠肝脏HBsAg组化显示密集成片的阳性细胞。实验组和对照组表达差异显著（P〈0.05）。结论将人脐血干细胞移植裸鼠肝脏内可以存活并分化成人肝细胞形成嵌合鼠,并能被HBV感染。     

High Incidence of Scrotal Hernias in a Closed Colony of FVB Mice

Although inguinal hernias are rarely reported to occur in mice, a high incidence of scrotal hernias was observed in a closed breeding colony of FVB/N mice. Unilateral or bilateral hernias occurred in more than 20% of the male mice in the colony that were available for necropsy over 3 inbred and 1 outcross generations; no female mice were affected. Organs commonly present within the hernial sac included the cecum and seminal vesicles. Hernias did not adversely affect the fertility or lifespan of the affected male mice. Although the condition was heritable, no clear pattern of transmission was evident.During development, the testes descend from the abdominal cavity through the inguinal canal and into the scrotum, guided by the processus vaginalis. In primates and carnivores, the processus vaginalis is largely or entirely obliterated during late gestation.¹³ When correct closure fails to occur, a hernial sac may travel through the deep inguinal ring to create an inguinal hernia.¹⁶ Protrusion of a hernial sac containing abdominal organs into the scrotum results in scrotal hernia, a severe and potentially dangerous form of inguinal hernia. In rodents, the inguinal canal is very short and the processus vaginalis remains patent throughout life, allowing the testes to pass freely between the scrotum and abdomen.¹³ Despite this potential pathway for herniation of abdominal organs, scrotal hernias have rarely been reported to occur in laboratory mice.Spontaneous inguinal hernias have rarely been reported in either male or female mice. Inguinal hernias occur in intact but not castrated male mice treated with estrogenic compounds and in intact female mice treated with testosterone or bearing testicular grafts.^1,2,10 Both male and female C57BL/6 mice that fail to express fibulin 3 develop multiple large hernias, including inguinal hernias.¹⁴ In these mice, herniation occurs at the myopectineal orifice, through the external inguinal ring. Female mice transgenic for insulin-like factor 3 develop inguinal hernias with 100% penetrance.¹¹ A recent report described a high incidence of lateral femoral hernias in an inbred colony of FVB/NHsd mice; the condition predominantly affected female mice.¹⁵ This phenomenon was attributed to genetic drift in a closed colony. We now report on another situation in which many hernias were noted in inbred FVB/N mice. In the present case, scrotal hernias occurred in a high proportion of FVB/N mice maintained in a closed breeding colony.     

Burst abdomen and incisional hernia: a prospective study of 1129 major laparotomies.

Burst abdomen and incisional herniation are continuing problems for the general surgeon. A prospective study was carried out to define the extent of the problem. Over five years from 1975 to 1980 a total of 1129 major laparotomy wounds in adults were assessed at regular intervals for 12 months after operation. There were 19 burst abdomens (1.7%) and 84 incisional hernias (7.4%). The introduction of the mass-closure technique reduced the incidence of burst abdomen from over 3% in 1975 to 0.95% in 1979. It did not, however, improve the rate for incisional hernias, which was 7.6% in 1979. Many factors are associated with incisional herniation: old age, male sex, obesity, bowel surgery, type of suture, chest infection, abdominal distension, and, most important, wound infection. More work is needed to find the ideal method of wound closure, and efforts should be made to eliminate wound infection.     

Bilateral Hernias in the Female

An experience with 216 bilateral hernias in female patients is reviewed. The condition is rare, occurring only once in every 250 patients admitted for a hernia repair. Bilateral primary indirect inguinal hernias were the most frequent type. Bilateral primary femoral hernias were quite rare while bilateral primary direct inguinal hernias were even more uncommon. Other rare bilateral combinations are briefly described. The incidence in children is given.Etiological factors are discussed, emphasizing the strong posterior wall of the inguinal canal in females.Two per cent of patients developed a recurrent hernia; one per cent of hernias recurred. No recurrence following a bilateral primary indirect inguinal hernia repair and no “femoral” recurrence following inguinal repair were recorded.     

Sliding indirect inguinal hernia

From 2 per cent to 5 per cent of all indirect inguinal hernias are of the sliding variety. (Sliding hernias are those in which part of the wall of the sac is formed by a viscus.) The proportion of sliding hernias is even higher in the aged. Hernias of this kind are found almost exclusively in males and usually on the left side. Preoperative diagnosis is not essential if the surgeon can recognize the lesion at operation and knows how to repair it properly. The LaRoque technique in which the peritoneal cavity is entered above the internal ring allows accurate definition of the pathological anatomy and effective repair of the hernia. It should be used in all true sliding indirect inguinal hernias.     

Bilateral inguinal hernia in a pig-tailed monkey (Macaca nemestrina).

A pig-tailed monkey developed bilateral inguinal hernias following escape from its cage and subsequent recapture. The hernias were surgically repaired and the monkey recovered.     

Herniation of gravid uterus: report of 2 cases and review of literature

Anterior abdominal wall hernias are uncommon, and herniation of a gravid uterus into these hernias is even rarer. Although reducible initially, the herniation of pregnant uterus may be complicated by incarceration and subsequent strangulation within the hernial sac, late in the course of pregnancy. There is no consensus over the management of this rare condition. Each case should be individualized. If uncomplicated, a conservative approach until term followed by delivery and herniorrhaphy is a good option. Here, 2 cases of herniation of gravid uterus into the anterior abdominal wall are described along with a brief review of literature pertaining to its presentation, complications, and management.     

Lateral Femoral Hernias in a Line of FVB/NHsd Mice: A New Confounding Lesion Linked to Genetic Background?

Several strains of transgenic mice derived from an inbred FVB/NHsd colony developed large masses on 1 or both flanks. Although originally suspected to be a phenotypic anomaly related to genetic modifications, nontransgenic littermates subsequently were affected with equal frequency, inculpating the FVB/NHsd founder colony. The masses were subcutaneous, soft, and exophytic and appeared over the course of a few weeks. Female mice were affected more frequently than males. Gross examination revealed the masses to consist of uni- or bilateral hernias of variable size, occasionally containing small or large intestine (or both), cecum, mesenteric adipose tissue, male reproductive organs, and ureters. All hernial sacs pouched through the femoral triangle laterally to the femoral vessels and therefore were classified as lateral femoral hernias. Lateral femoral hernias have not previously been described in the veterinary literature and have never been described as background lesions in a strain of mice. Our findings suggest likely genetic drift in this strain of FVB/NHsd mice, causing a background lesion that confounded phenotypic analyses of transgenic mice derived from this strain.     

Introduction of the human AVPR1A gene substantially alters brain receptor expression patterns and enhances aspects of social behavior in transgenic mice

Central arginine vasopressin receptor 1A (AVPR1A) modulates a wide range of behaviors, including stress management and territorial aggression, as well as social bonding and recognition. Inter- and intra-species variations in the expression pattern of AVPR1A in the brain and downstream differential behavioral phenotypes have been attributed to differences in the non-coding regions of the AVPR1A gene, including polymorphic elements within upstream regulatory areas. Gene association studies have suggested a link between AVPR1A polymorphisms and autism, and AVPR1A has emerged as a potential pharmacological target for treatment of social cognitive impairments and mood and anxiety disorders. To further investigate the genetic mechanism giving rise to species differences in AVPR1A expression patterns and associated social behaviors, and to create a preclinical mouse model useful for screening drugs targeting AVPR1A, we engineered and extensively characterized bacterial artificial chromosome (BAC) transgenic mice harboring the entire human AVPR1A locus with the surrounding regulatory elements. Compared with wild-type animals, the humanized mice displayed a more widely distributed ligand-AVPR1A binding pattern, which overlapped with that of primates. Furthermore, humanized AVPR1A mice displayed increased reciprocal social interactions compared with wild-type animals, but no differences in social approach and preference for social novelty were observed. Aspects of learning and memory, specifically novel object recognition and spatial relocation recognition, were unaffected. The biological alterations in humanized AVPR1A mice resulted in the rescue of the prepulse inhibition impairments that were observed in knockout mice, indicating conserved functionality. Although further behavioral paradigms and additional cohorts need to be examined in humanized AVPR1A mice, the results demonstrate that species-specific variations in the genomic content of regulatory regions surrounding the AVPR1A locus are responsible for differential receptor protein expression patterns across species and that they are likely to contribute to species-specific behavioral variation. The humanized AVPR1A mouse is a potential preclinical model for further understanding the regulation of receptor gene expression and the impact of variation in receptor expression on behaviors, and should be useful for screening drugs targeting human AVPR1A, taking advantage of the expression of human AVPR1A in human-relevant brain regions.KEY WORDS: AVPR1A, Humanized mouse, Social behavior, Species-specific, Microsatellite, Autism