阿托伐他汀对Lewis大鼠实验性自身免疫性心肌炎作用的研究

目的： 探讨阿托伐他汀对实验性自身免疫性心肌炎(EAM）大鼠Th1/Th2偏离的影响及对EAM的治疗价值。方法: 6-8周雄性Lewis大鼠31只，其中8只作为正常对照；23只以猪心肌肌球蛋白免疫制成EAM模型，免疫后随机分为阿托伐他汀大剂量（10 mg·kg^-1·d^-1）组、小剂量（1 mg·kg^-1·d^-1）组和未治疗组，连续用药 21 d。第 21 d，行超声心动图检测；取心肌组织，观察大体及镜下炎症程度；ELISA检测血浆IL-2、IL-4、IL-10及IFN-γ等细胞因子水平,并以IFN-γ/IL-4比值作为Th1/Th2偏离方向指标。结果: 阿托伐他汀使EAM大鼠心室肥厚减轻，LVEDd降低，射血分数增加；心脏重量/体重比值及炎症程度分级显著降低；Th1型细胞因子(IFN-γ, IL-2)水平降低，Th2型细胞因子水平(IL-4, IL-10)升高。3组间TC、TG及HDL-C水平未见明显差异。结论: 阿托伐他汀使Th1/Th2平衡向Th1方向偏离，抑制EAM炎症反应。表明阿托伐他汀的免疫调节效应及在自身免疫病治疗中的应用前景。     

多种金属离子与单宁酸反应媒染微血管的对比研究

目的　镜下观察单宁酸与金属盐溶液中Ca2 + 、Au+ 、Ag+ 、Pb2 + 、Cu2 + 、Al3 + 、U6+ 、K+ 联用显示大脑微血管的效果。方法　用单宁酸媒染固定液灌流大鼠 ,取脑切片 ,入氯化钙、氯化金、硝酸银、硝酸铅、硫酸铜、硫酸铝钾、醋酸双氧铀、高锰酸钾和重铬酸钾等溶液中呈色。结果　单宁酸与Ca2 + 、Cu2 + 、Ag+ 、Al3 + 结合显示血管清晰 ,与Au+ 、U6+ 、Pb2 + 、K+ 联用媒染血管效果欠佳。结论　含Ca2 + 、Cu2 + 、Ag+ 、Al3 + 的金属盐类可替代氯化铁媒染微血管 ,氯化金、醋酸双氧铀、硝酸铅、重铬酸钾和高锰酸钾不宜与单宁酸联用来显示血管     

人α2（I）型胶原基因启动子活性研究

目的 探索器官纤维化形成中调控I型胶原基因高水平转录的启动片段及TGF-β、PDGF－BB、IGF－1等细胞因子对其活性的影响。方法 从人α2（I）胶原基因转录起始点上游－2.4kb至＋58bp的片段中，取长度不等的片段作为启动子与含氯霉素乙酰基转移酶（CAT）报告基因的质粒组成5个重组体，转染上述重组体至正常人原代培养皮肤成纤维细胞，测定细胞CAT表达水平以比较各重组体的启动子活性，同时加入细胞因子，以测定其对I型胶原启动序列的影响。结果 除－129～＋58bp序列外，其余4个重组体CAT表达水平均较高，其中－2292～＋58bp、－1476～ 58bp序列具较强启动CAT表达活性，－339～ 58bp、－616～ 58bp片段次之。TGF－β、IGF－1均能在一定程度上调人α2（I）胶原基因启动活性。结论 人α2（I）胶原基因片段－2292～ 58bp、－1476～ 58bp、－339～ 58bp有高启动活性，是进一步研究纤维化相关DNA结合蛋白的重要调控靶序列。TGF－β、IGF－1促进胶原表达，与其上调胶原基因启动活性有关。     

A female with X‐linked Nephrogenic diabetes insipidus in a family with inherited central diabetes Insipidus: Case report and review of the literature

There are two forms of diabetes insipidus, central (neurohypophyseal), and nephrogenic, caused by pathogenic variants in the AVP gene and the AVPR2 or AQP2 genes, respectively. We report on a four‐generation family, seven individuals had central diabetes insipidus (CDI) and the female index patient seen from age 16 to 26 years had (mild) nephrogenic diabetes insipidus. In her father with CDI, a known pathogenic heterozygous AVP variant c.232_234del p.(Glu78del) was identified, confirming the diagnosis of CDI in him and the other affected family members. In the proband, molecular analysis disclosed a novel heterozygous AVPR2 gene variant, c.962A > T p.(Asn321Ile) and an extremely skewed X‐inactivation, confirming X‐linked nephrogenic diabetes insipidus (XL‐NDI). Whole exome sequencing showed no further causative mutation. This is the first report on the co‐existence of CDI and NDI in one family. Our review of symptomatic female AVPR2 heterozygotes includes 23 families with at least one affected female (including this study). There were 21 different causative mutations. Mutation types in females did not differ from those in males. Both severe XL‐NDI and mild forms were reported in females. All six females with severe XL‐NDI had complete loss‐of‐function (null) mutations. The remaining 17 female probands had milder XL‐NDI caused by 14 missense variants and three null variants of the AVPR2 gene. X‐inactivation was studied in nine of these females; all showed extreme or slight skewing. The review underlines that XL‐NDI in female AVPR2 heterozygotes is always accompanied by skewed X‐inactivation, emphasizing a need for X‐inactivation studies in these females.     

以报告基因荧光素酶研究HPRE功能元件与IFN-α应答的关系

目的:以荧光素酶(luciferase,LUC)基因为报告基因,探讨HBV转录后调节序列(HPRE)的功能元件(α、β1和β2)对IFNα作用的影响。方法:用PCR法从载体pGEMluc中扩增LUC基因,并克隆入真核表达载体pcDNA3.0中。以含乙肝病毒(HBV)基因组的质粒A01为模板,扩增HPRE(完整的HPRE)、HPREαβ1及HPREβ1β2片段,并分别插入LUC基因的下游。利用脂质体介导的转染法,将重组质粒分别导入人肝癌细胞株HepG2中,用LUC检测系统检测IFNα作用前后LUC表达活性的变化。结果:经测序证实,重组质粒pcDNA3.0luc、pcDNA3.0lucHPRE、pcDNA3.0lucHPREαβ1及pcDNA3.0lucHPREβ1β2构建成功。检测结果显示,IFNα作用前,HPRE、HPREαβ1和HPREβ1β2均能提高LUC的活性,IFNα作用后,HPRE和HPREβ1β2能明显降低LUC的活性,而HPREαβ1对其表达则没有显著影响。结论:HPRE的功能元件β2与IFNα作用的关系最为密切,而功能元件α和β1在IFNα应答中作用甚小,提示IFNα诱导产生的HPRE抑制性结合蛋白很可能是与β2结合的,为进一步研究IFNα在治疗HBV感染和慢性肝炎中的作用机制,以及HPRE抑制性结合蛋白的作用提供了一定的实验依据。     

Comparative morphological differences between umbilical cords from chronic hypertensive and preeclamptic pregnancies

To compare morphological changes in the umbilical cords from chronic hypertensive and preeclamptic patients having normal or pathological umbilical artery Doppler ultrasonographic results. Umbilical cords from 34 normotensive, 31 chronic hypertensive and 70 preeclamptic women with normal and abnormal Doppler flow velocity waveforms (FVW) at 35-40 gestational weeks were studied. Morphological changes in the umbilical cords were examined on formalin-fixed, paraffin-embedded sections. The total umbilical cord area, total vessel area, and wall thickness of umbilical vessels were measured in systematic random samples using unbiased stereology methods. An ANOVA test was used for statistical analysis. In the chronic hypertensive and preeclamptic groups with normal Doppler FVW, the thickness of the umbilical cord vessels remained nearly constant, whereas both the total area and the lumen area were reduced. These changes correlate with the histopathological findings, suggesting a mainly vasoconstrictive effect. By contrast, analysis of the preeclamptic group with pathologic Doppler FVW showed a comparable reduction of all parameters of the umbilical cord. Histopathological findings were related to smaller, contracted smooth muscle cells of the vessel wall, which is suggestive of a predominant hypoplastic mechanism. As a result of reduced uteroplacental perfusion, fetal hypoxia and intrauterine growth retardation become unavoidable in preeclampsia. The histopathological changes in the umbilical cord between the chronic hypertensive and preeclamptic patients depend on the Doppler results. In conclusion, the umbilical artery Doppler FVW indices provide good values for predicting intrauterine growth retardation in preeclamptic patients.     

NOD小鼠胸腺细胞TCR介导的信号通路缺陷

目的 进一步研究NOD小鼠T细胞应答改变机理。方法 用抗TCR抗体、ConA激活NOD小鼠胸腺细胞，分析TCR介导的信号通路的水平。结果 与Balb／c小鼠胸腺细胞相比，抗TCR抗体诱导的增殖应答较弱，与年龄及NOD胸腺CD4^ CD8^-和CD4^-CD8^ SP细胞有关；rIL-2能部分恢复对TCR抗体应答的缺乏。NOD小鼠对PMA IONO和PMA anti—TCR-mAb应答正常，但对anti-TCRmAb IONO应答缺乏。结论 与年龄有关的NOD小鼠胸腺细胞对TCR抗体应答的缺乏与T细胞激活时上游PKC信号通路的缺乏有关。     

Galectin-3基因在人孕早期绒毛和蜕膜中的表达及与妊娠的关系

目的 研究人早孕绒毛和子宫蜕膜中galectin-3基因mRNA和蛋白的表达，探讨其与滋养细胞浸润的关系。方法 采用半定量RT—PCR和Western-blot分别对70例（分为7个孕周组，每组10例）正常早孕绒毛和蜕膜中gal-3mRNA和蛋白表达水平进行检测 结果 在整个孕早期绒毛和子宫蜕膜中均有gal-3基因表达，且在绒毛和蜕膜中随着孕周的增加其表达总体呈上升趋势，两者表达趋势一致。结论 gal-3基因在滋养细胞和蜕膜表达中随孕龄增长而呈上调趋势，提示该基因在调控早孕滋养细胞的浸润行为中起重要的作用。     

人胚胎干细胞原代克隆生长及其传代的研究

目的 评价人胚胎干细胞建系与囊胚质量、原代克隆生长的关系。方法 D3废弃胚胎成组共培养获得不同质量的囊胚，免疫刀去除囊胚滋养外胚层细胞后，将内细胞团(ICM)接种到饲养细胞层上生长、传代。结果 从质量好的囊胚得到的人胚胎干细胞传代的代数更多；原代克隆生长快的人胚胎干细胞传代效率更高。结论 人胚胎干细胞建系与囊胚质量、原代克隆生长情况密切相关。