An efficient SNP system for mouse genome scanning and elucidating strain relationships

A set of 1638 informative SNP markers easily assayed by the Amplifluor genotyping system were tested in 102 mouse strains, including the majority of the common and wild-derived inbred strains available from The Jackson Laboratory. Selected from publicly available databases, the markers are on average ~1.5 Mb apart and, whenever possible, represent the rare allele in at least two strains. Amplifluor assays were developed for each marker and performed on two independent DNA samples from each strain. The mean number of polymorphisms between strains was 608±136 SD. Several tests indicate that the markers provide an effective system for performing genome scans and quantitative trait loci analyses in all but the most closely related strains. Additionally, the markers revealed several subtle differences between closely related mouse strains, including the groups of several 129, BALB, C3H, C57, and DBA strains, and a group of wild-derived inbred strains representing several Mus musculus subspecies. Applying a neighbor-joining method to the data, we constructed a mouse strain family tree, which in most cases confirmed existing genealogies.     

艾芬地尔干预后氯化锂-匹罗卡品致(癎)大鼠的慢性期脑电图特征

目的:了解艾芬地尔干预对氯化锂(LiCl)-匹罗卡品(Pilo)致(癎)大鼠脑电图随时间变化的特征.方法:通过腹腔注射LiCl-Pilo建立癫(癎)动物模型.60只雄性Wistar大鼠分为对照组、模型组和艾芬地尔干预组,分别在造模后第7、15、30、60天四个时间点观察各组大鼠行为、脑电图改变,每一个时间点5只大鼠.结果:对照组无NFDA1性发作,经过4-20天的潜伏期后,干预组及模型组的脑电图波幅及频率骤然减低,在慢性期干预组较模型组的波幅逐渐增高,频率逐渐增快,但仍未达发作当时的水平,且两组间比较差异无统计学意义.干预组较模型组更早更快趋于正常化.结论:艾芬地尔在LiCl-Pilo致(癎)大鼠模型中具有抗惊厥作用,慢性期脑电图有特征性改变.     

尿VEGF、ET检测对2型糖尿病患者早期肾脏微血管损伤的诊断价值

目的探讨尿血管内皮生长因子（VEGF）、内皮素（ET）对2型糖尿病患者早期肾脏微血管损伤的诊断价值。方法按尿微量蛋白排泄率把76例2型糖尿病患者分为无肾病、早期肾病、临床肾病3组。采用放射免疫法（RIA）和酶联免疫吸附试验（EHSA）测定尿VEGF、α1-微球蛋白（α1-MG）、β2-微球蛋白（β2-MG）、免疫球蛋白G（IgG）、ET水平。结果2型糖尿病患者尿VEGF、ET与对照组比较差异有统计学意义（t≥2．29，P〈0．01），且其水平随病情加重而明显升高；与α1-MG、β2-MG、IgG、Alb、糖化血红蛋白（HbAlc）呈正显著相关（r≥0．309）．而与空腹血糖（FBG）无明显相关（r≤0．06）。结论VEGF、ET参与糖尿病微血管病变的发生、发展；尿VEGF、ET是诊断2型糖尿病患者早期肾脏微血管损伤的敏感指标，对糖尿病肾病的早期诊断和肾功能损害程度的判断有一定的临床诊断价值。     

上皮间质转化(EMT)及其分子机制

上皮细胞受到细胞外因子刺激后向间质细胞转化的现象与肿瘤的浸润转移密切相关。在此过程中上皮细胞的极性丧失，迁移和运动能力增强，同时上皮表型丢失而逐渐获得间质表型。参与这一过程的细胞内信号转导途径有受体酪氨酸激酶Ras-MAPK途径，Src激酶，Rho家族激酶，PI3K/AKT途径，Wnt信号通路，转录因子等。     

鸟苷素家族研究进展

Guanylin family, described in recent 10 years, is a series of small peptides (including guanylin, uroguanylin and lymphoguanylin) with structural and functional similarities to heat- stable entero-toxins (STs) elaborated by various pathogenic bacteria. They are abundance of cysteines and are endogenous activators of guanylyl cyclase - C (GC - C) receptors. Immunoreactive guanylin family peptides are localized in many human organs and tissues, especially in gastrointestinal tract and kidney, and play an important role in regulation of water and salt homeostasis. Recent studies showed that the mRNA levels of guanylin family peptides were down - regulated in colorectal cancers; oral intake of uroguanylin might suppress polyp formation in Apc(Min/ ) mouse, and 111^In-labeled-ST peptide analog might specifically target human colon can-cers. These evidences highlight that guanylin family may have a potential application in diagnosis and therapy effects of colorectal cancers.     

白蛋白微泡促进报告基因在细胞表达的实验研究

目的: 探讨白蛋白微泡作为非病毒载体在基因传输中的作用。方法:6孔板培养脐静脉内皮细胞(EC)和血管平滑肌细胞(VSMC),每孔加pcDNA3.1/His/LacZ质粒20 μg,加不同浓度微泡或不加微泡,超声条件为连续波,频率2MHz,机械指数1.8,照射时间1 min,48 h后计算蓝染细胞百分率和β-半乳糖苷酶活性。另以不同浓度微泡及超声照射时间处理细胞,测定细胞增殖情况。结果:与单纯超声质粒组相比,含微泡组蓝染细胞率增加约10-15倍(其中内皮细胞组11.6倍,平滑肌细胞组15.2倍),报告基因表达定量增加近8倍。微泡浓度为10％时细胞转染效率最高。超声照射对细胞增殖无影响,微泡浓度为50％时有明显的细胞毒作用。结论:白蛋白微泡在超声作用下能明显增加基因的传输效率,有可能成为一种安全有效的基因治疗的载体。     

人癌胚抗原重组痘苗病毒转染树突状细胞体外诱导的抗肿瘤免疫

目的：研究人癌胚抗原重组痘苗病毒（rV-CEA)转染外周血树突状细胞（DC）后在外体诱导的抗CEA分泌性肿瘤免疫。方法：分离晚期结肠癌患者的外周血单核细胞，在体外用人重组粒－巨噬细胞集落刺激因子（GM－CSF）及白细胞介素4（IL－4）培养成DC，再用rV-CEA转染DC后激发自体T细胞，观察其体外激发自体T细胞的增殖能力及其诱导的细胞毒性T淋巴细胞（CTL）对自体CEA分泌性肿瘤细胞的杀伤活性，并与野生型痘苗病毒（W－VV）及无病毒转染的DC所激发的T细胞进行比较。结果：经rV-CEA转染的DC能显著刺激自体T细胞的增殖，其激活的T细胞对CEA分泌性自体肿瘤细胞具有特异性杀伤作用。结论：rV-CEA转染的DC可以诱导体CEA分泌性肿瘤免疫。     

Gene therapy for pituitary tumors

Pituitary tumors are the most common primary intracranial neoplasms. Although most pituitary tumors are considered typically benign, others can cause severe and progressive disease. The principal aims of pituitary tumor treatment are the elimination or reduction of the tumor mass, normalization of hormone secretion and preservation of remaining pituitary function. In spite of major advances in the therapy of pituitary tumors, for some of the most difficult tumors, current therapies that include medical, surgical and radiotherapeutic methods are often unsatisfactory and there is a need to develop new treatment strategies. Gene therapy, which uses nucleic acids as drugs, has emerged as an attractive therapeutic option for the treatment of pituitary tumors that do not respond to classical treatment strategies if the patients become intolerant to the therapy. The development of animal models for pituitary tumors and hormone hypersecretion has proven to be critical for the implementation of novel treatment strategies and gene therapy approaches. Preclinical trials using several gene therapy approaches for the treatment of anterior pituitary diseases have been successfully implemented. Several issues need to be addressed before clinical implementation becomes a reality, including the development of more effective and safer viral vectors, uncovering novel therapeutic targets and development of targeted expression of therapeutic transgenes. With the development of efficient gene delivery vectors allowing long-term transgene expression with minimal toxicity, gene therapy will become one of the most promising approaches for treating pituitary adenomas.     

Crouzon综合征基因突变检测

目的 研究1个Crouzon综合征家系及1例散发的Crouzon综合征患者的成纤维生长因子受体2(fibroblast growth factors receptor 2,FGFR2)基因突变情况.方法 在1个Crouzon综合征家系的10名成员,和另一例散发者的外周血提取基因组DNA,PCR扩增FGFR2基因的第8和10外显子(部分家族成员仅扩增第8外显子),产物纯化后直接进行DNA测序检测突变.结果 家系中3名成员及另1例散发者FGFR2基因第8外显子的833位核苷酸发生G→T的转换突变,该突变为错义突变,使该位点所编码的氨基酸由半胱氨酸变为苯丙氨酸(C278F).该突变为杂合子突变.结论 FGFR2基因突变是Crouzon综合征致病原因.     

胎牛血清外泌体对成骨细胞增殖的作用

背景:研究表明外泌体具有促进骨再生的能力,但从胎牛血清中提取的外泌体是否可以促进骨形成仍存在争议。目的:观察胎牛血清外泌体对成骨细胞增殖能力的影响,从而为临床治疗骨破坏提供新思路。方法:通过超速离心法从胎牛血清中提取外泌体,采用透射电子显微镜和Western blot法验证外泌体是否提取成功;然后用10 mg/L胎牛血清外泌体干预成骨前体细胞MC3T3-E1,通过CCK-8实验检测外泌体对成骨细胞增殖能力的影响,Western blot检测外泌体对成骨细胞骨形态发生蛋白2和骨桥蛋白表达的影响。以不含外泌体的胎牛血清培养的MC3T3-E1细胞为对照组。结果与结论:①胎牛血清外泌体具有典型的脂质双层膜结构,大小在30-150 nm之间,外泌体表面标记因子CD81表达呈阳性,而微囊表面标记物CD40表达呈阴性;②外泌体组的增殖能力明显高于对照组,差异有显著性意义(P<0.05);外泌体组成骨标志性因子骨形态发生蛋白2和骨桥蛋白的表达水平明显高于对照组,差异有显著性意义(P<0.05);③结果表明,胎牛血清外泌体对成骨细胞的增殖起促进作用,可为临床治疗骨破坏提供新思路。