脆性X综合征中不稳定的DNA序列和异常甲基化研究

采用ＰＣＲ结合序列胶分析的方法，对８２条正常中国人Ｘ染色体ＦＲＡＸＡ位点（ＣＧＧ）ｎ重复序列拷贝数的多态性进行了测定，其范围为２１～３１，高峰为２７。并通过Ｓｏｕｔｈｅｒｎ杂交分析了来自６个Ｆｒａ（Ｘ）家系的１５名成员（ＣＧＧ）ｎ的拷贝数与该重复序列上游ＣｐＧ岛的甲基化状态。Ｆｒａ（Ｘ）患者（ＣＧＧ）ｎ大量扩增并伴随ＣｐＧ岛异常甲基化。Ｆｒａ（Ｘ）携带者女性（ＣＧＧ）ｎ扩增较少，有嵌合现象。其子代或产生更大扩增，或保持原来状态，呈动态突变遗传特征。     

Serum Levels of Free Insulin-Like Growth Factor (IGF)-I in Normal Children

Serum levels of free insulin-like growth factor (IGF)-I were measured by immunoradiometric assay (IRMA) in fasting sera of 137 normal boys and 120 normal girls aged from 8 to 15 yr to study relationships between free IGF-I levels and ages, total IGF-I, IGF binding protein (IGFBP)-1, IGFBP-3, and acid-labile subunit (ALS) levels. In both sexes, serum free IGF-I levels and the ratios of free IGF-I to total IGF-I were significantly higher in the pubertal age groups than in the prepubertal age groups. Serum levels of free IGF-I showed a significant positive correlation with those of total IGF-I, IGFBP-3 and ALS, while they showed a significant negative correlation with those of IGFBP-1. These observations suggest that increase in serum free IGF-I levels during puberty is caused by a dramatic increase in total IGF-I, rather than IGFBP-3, and a decrease in IGFBP-1. Also, high free IGF-I levels may play an important role in pubertal growth spurt.     

The evolution of a neo-XY1Y2 sex chromosome system by autosome–sex chromosome fusion in Dundocoris nodulicarinus Jacobs (Heteroptera: Aradidae: Carventinae)

Sibling subspecies of Dundocoris nodulicarinus, inhabiting different isolated indigenous evergreen forests in South Africa, have chromosome numbers of 2n(male) = 14XY, 9XY1Y2 and 7XY1Y2. The ancestral chromosome number of Dundocoris is probably 2n(male) = 28XY and several chromosome fusions were involved in the karyotype evolution of these taxa. The XY1Y2 sex chromosome system of the 9XY1Y2 D. nodulicarinus novenus originated by the fusion of a large autosome with the X-chromosome, forming a neo-X with the homologue of the fused autosome forming the neo-Y (=Y1) and the original Y-chromosome, the Y2. While the original X- and Y-chromosomes are heterochromatic and heteropycnotic during prophase I, the autosomal part of the neo-X and the neo-Y stay euchromatic and behave like a normal autosomal pair, forming synapsis and chiasmata. The XY1Y2 sex chromosome system of the 7XY1Y2 D. nodulicarinus septeni probably originated from the 9XY1Y2 karyotype when the homologous chromosomes of a small autosomal pair fused with the original X- and Y-chromosomes, respectively. In both the subspecies with the neo-XY1Y2 systems, the original sex chromosomes still undergo chromatid segregation at anaphase I (= post-reductional). The evolution and behaviour of the karyotypes and sex chromosome systems during the course of meiosis in the subspecies of D. nodulicarinus are described, discussed and illustrated.     

3D kinematic analysis of the acromioclavicular joint during arm abduction using vertically open MRI.

Many researchers have evaluated the motions of the shoulder girdle, especially scapular and humeral motion. However, few reports exist that describe motions of the acromioclavicular joint. The purpose of the present study was to analyze the 3D kinematics of the acromioclavicular joint during arm abduction using 3D MR images obtained by a vertically open MRI. Fourteen shoulders of seven volunteers were examined in seven static positions from 0 degrees to the maximum abduction in a seated position. 3D surface models of the clavicle and scapula were created, and the movements of the acromioclavicular joint from 0 degrees to each position were calculated using the volume-based registration technique. From these calculations, the translations were evaluated and the rotational motions were analyzed using the concept of the screw axis. In the anteroposterior direction, the clavicle translated most posteriorly (-1.9 +/- 1.3 mm) at 90 degrees of abduction and most anteriorly (1.6 +/- 2.7 mm) at maximum abduction. In the superoinferior direction, the clavicle translated slightly superiorly (0.9 +/- 1.9 mm). When analyzing relative motion of the scapula with respect to the clavicle, the scapula generally rotated about a specific screw axis passing through the insertions of both the acromioclavicular and the coracoclavicular ligaments on the coracoid process. The average rotation was 34.9 +/- 8.4 degrees.     

α_1肾上腺素能受体阻滞剂萘哌地尔治疗慢性非细菌性前列腺炎的临床研究

目的:探讨α1肾上腺素能受体阻滞剂萘哌地尔(Naftopidil)治疗慢性非细菌性前列腺炎的有效性及安全性。方法:采用开放、自身对照、多中心的临床试验方法,应用萘哌地尔25mg,每日1次,对106例慢性非细菌性前列腺炎(NBP)患者进行了为期4周的治疗。以美国国立卫生院慢性前列腺炎症状评分(NIHCPSI)、前列腺液(EPS)WBC计数及最大尿流率(MFR)为疗效指标,对其有效性及安全性进行观察。结果:服药4周后,可评价病例105例。全组患者NIHCPSI总评分治疗前后平均减低12.0分(P<0.001),症状评分平均减低7.9分(P<0.001),生活质量评分平均减低4.1分(P<0.001)。EPS中WBC计数治疗前及治疗后分别为(15.2±15.1)、(9.5±12.0)个/HP(P<0.01)。MFR治疗前及治疗后分别为(19.2±4.8)、(22.7±4.9)ml/s(P<0.01)。按症状改善评价,治愈2例(1.9%),显效32例(30.5%),有效55例(52.4%),无效16例(15.2%)。总显效率为32.4%,总有效率为84.8%。3例有轻度头晕,1例食欲不佳,不良事件发生率3.81%。结论:萘哌地尔治疗慢性非细菌性前列腺炎安全、有效。     

NPHS1两种新突变蛋白分子的细胞内分布

目的：研究NPHS1两种新突变编码蛋白在细胞内的分布与先天性肾病综合征发病机制的关系。方法：构建野生型和两种突变型NPHS1克隆，并转染至COS7细胞内，应用免疫荧光双标记的方法，分别进行细胞内及细胞表面的荧光标记，通过共聚焦显微镜对裂隙膜分子Nephrin在细胞内的分布进行研究。结果：野生型Nephrin表现为细胞内和细胞膜染色模式；V822M和C265R则主要为细胞内内质网染色模式，细胞膜着色几乎缺失。结论：突变的Nephrin蛋白由于错误折叠，不能由内质网被输送至细胞表面，这可能是先天性肾病综合征发病的机制之一。     

持续吸入不同浓度氧气对新生大鼠肺血管内皮生长因子及其受体tuRNA表达的影响

目的 探讨持续吸入不同浓度氧气对新生大鼠肺血管内皮生长因子(VEGF)及其受体1(VEGFRl)和受体2(VEGFR2)mRNA表达的影响.方法 新生足月SD大鼠32只,随机分为对照组和实验组.实验组生后12 h开始持续吸入氧气,按不同的吸入氧浓度,将实验组又分为30%O2组、50%O2组和75%O2组.对照组吸入空气.每组8只.于实验开始后21 d处死实验大鼠,取出右肺下叶,RT-PCR技术检测VEGF、VEGFR1和VEGFR2 mRNA表达,根据2-△△CT的计算方法,实验组基因表达差异用实验组相对于对照组基因表达量的倍数表示.结果 与对照组相比,30%O2对新生大鼠肺VEGF及其受体mRNA表达无影响.75%O2组VEGF mRNA表达是对照组的0.48倍;50%O2组、75%O2组VEGFR1 mRNA分别为对照组的0.18倍和0.06倍;VEGFR2 mRNA分别为对照组的0.22倍和0.10倍,差异均有统计学意义(P<0.05).结论 长时间吸入低浓度氧对新生大鼠肺VEGF及其受体mRNA影响不明届,而持续吸入中等浓度及较高浓度氧可降低VEGF及其受体mRNA的表达.     

Mapping corpus callosum deficits in autism: an index of aberrant cortical connectivity.

BACKGROUND: Volumetric studies have reported reductions in the size of the corpus callosum (CC) in autism, but the callosal regions contributing to this deficit have differed among studies. In this study, a computational method was used to detect and map the spatial pattern of CC abnormalities in male patients with autism. METHODS: Twenty-four boys with autism (aged 10.0 +/- 3.3 years) and 26 control boys (aged 11.0 +/- 2.5 years) underwent a magnetic resonance imaging (MRI) scan at 3 Tesla. Total and regional areas of the CC were determined using traditional morphometric methods. Three-dimensional (3D) surface models of the CC were also created from the MRI scans. Statistical maps were created to visualize morphologic variability of the CC and to localize regions of callosal thinning in autism. RESULTS: Traditional morphometric methods detected a significant reduction in the total callosal area and in the anterior third of the CC in patients with autism; however, 3D maps revealed significant reductions in both the splenium and genu of the CC in patients. CONCLUSIONS: Statistical maps of the CC revealed callosal deficits in autism with greater precision than traditional morphometric methods. These abnormalities suggest aberrant connections between cortical regions, which is consistent with the hypothesis of abnormal cortical connectivity in autism.     

Analysis of Vascular Endothelial Growth Factor (VEGF) and a receptor subtype (KDR/flk-1) in the liver of rats exposed to riddelliine: a potential role in the development of hemangiosarcoma

Riddelliine alters hepatocellular and endothelial cell kinetics and function including stimulating an increase in hepatocytic vascular endothelial growth factor (VEGF) in the absence of increased serological levels of VEGF (Nyska etal. 2002). The objective of this study was to further assess hepatic VEGF and KDR/flk-1 synthesis and expression by hepatic cells under riddelliine treatment conditions. Forty-two male F344/N rats were dosed by gavage with riddelliine (0, 1.0, and 2.5 mg/kg/day) for 6 weeks. Seven animals/group were sacrificed after 8 consecutive daily doses; remaining rats were terminated after 30 daily doses, excluding weekends. Hepatic tissues were evaluated by immunohistochemistry and in situ hybridization. The results showed that VEGF mRNA expression was observed in control and treated animals; however, qualitative differences were noted. Treated animals exhibited VEGF mRNA in clustered, focal hepatocytes and bile duct epithelium, whereas VEGF mRNA in hepatocytes from vehicle control rats was distributed evenly across all hepatocytes. Results evaluating the distribution of the VEGF cognate receptor, KDR/flk-1 showed that randomly distributed, rare sinusoidal endothelium, including those demonstrating karyomegaly and cytomegaly expressed KDR/flk-1. Phosphorylation of KDR/flk-1 at pTyr996 and pTyr1054/1059, but not pTyr951, was also detected, evidence that endothelial cell KDR/flk-1 was activated. These results suggest that both hepatocytes and endothelial cells are targets of riddelliine-induced injury. We speculate that damage to both populations of cells may lead to dysregulated VEGF synthesis by hepatocytes and activation of KDR/flk-1 by endothelium leading to the induction of sustained endothelial cell proliferation, culminating in the development of hepatic hemangiosarcoma.     

慢性乙型肝炎患者APOBEC3G mRNA水平对肝组织损害的预测

目的 研究慢性乙型肝炎患者PBMC和活检肝组织的APOBEC3G(A3G)mRNA表达状况并探讨两者之间的相关性;研究A3G mRNA转录表达水平与血清HBV DNA、ALT、PT水平及乙型肝炎肝组织学活动度Knodell计分的相关性.方法 采用实时荧光相对定苗RT-PCR的方法 检测45例慢性乙型肝炎患者PBMC及肝组织中A3G mRNA的表达水平,同时采用实时荧光定量PCR方法 检测血清HBV DNA;常规检测TBil、ALT、PT及乙型肝炎肝组织学活动度Knodell计分.同时设15例健康体检者为阴性对照组.结果 ①慢性乙型肝炎患者PBMC、肝组织均表达A3G mRNA.PBMC A3G mRNA表达水平与活检肝组织A3G mRNA表达呈正相关(r=0.457,P<0.05);②PBMC A3G mRNA与肝组织炎症活动度呈负相关(r＝-0.441,P<0.05);③PBMC A3G表达水平与HBV DNA呈正相关(r=0.299,P<0.05),与TBil、ALT、PT无相关性.结论 本组研究显示:①体内研究慢性乙型肝炎患者A3G mRNA抗HBV作用,可首选外周血作为临床适用样本.②慢性乙型肝炎患者PBMC A3G mRNA水平可预测其肝组织损害程度,PBMC A3G mRNA水平越高,肝组织损害越轻.