Immunological parameters in girls with Turner syndrome

Disturbances in the immune system has been described in Turner syndrome, with an association to low levels of IgG and IgM and decreased levels of T- and B-lymphocytes. Also different autoimmune diseases have been connected to Turner syndrome (45, X), thyroiditis being the most common.     

A point mutation in the 5' splice site of the dystrophin gene first intron responsible for X-linked dilated cardiomyopathy

X-linked dilated cardiomyopathy (XLDC) is a familial heart disease presenting in young males as a rapidly progressive congestive heart failure, without clinical signs of skeletal myopathy. This condition has recently been linked to the dystrophin gene in some families and deletions encompassing the genomic region coding for the first muscle exon have been detected. In order to identify the defect responsible for this disease at the molecular level and to understand the reasons for the selective heart involvement, a family with a severe form of XLDC was studied. In the affected members, no deletions of the dystrophin gene were observed. Analysis of the muscle promoter, first exon and intron regions revealed the presence of a single point mutation at the first exon-intron boundary, inactivating the universally conserved 5' splice site consensus sequence of the first intron. This mutation introduced a new restriction site for MseI, which cosegregates with the disease in the analyzed family. Expression of the major dystrophin mRNA isoforms (from the muscle-, brain- and Purkinje cell-promoters) was completely abolished in the myocardium, while the brain- and Purkinje cell- (but not the muscle-) isoforms were detectable in the skeletal muscle. Immunocytochemical studies with anti- dystrophin antibodies showed that the protein was reduced in quantity but normally distributed in the skeletal muscle, while it was undetectable in the cardiac muscle. These findings indicate that expression of the muscle dystrophin isoform is critical for myocardial function and suggest that selective heart involvement in dystrophin- linked dilated cardiomyopathy is related to the absence, in the heart, of a compensatory expression of dystrophin from alternative promoters.      

Poly(D,L-lactide/epsilon-caprolactone)/hydroxyapatite composites as bone filler: an in vivo study in rats

In this study, a novel composite bone substitute was implanted in animal models (rats) and their in vivo characteristics were examined. A D,L-lactide and E-caprolactone copolymer (Mw: 80,000; Mn:40,000, and PI:2.00) was synthesized by ring-opening polymerization of the respective dimers using stannous octoate as the catalyst. The final ratio of D,L-lactide to epsilon-caprolactone obtained by 1NMR was 60/40. Hydroxyapatite (HA) powder was loaded in the copolymer. The HA/copolymer ratio was 60/40 (w/w). These composites were easily shaped by hand. Animal tests were performed on mature wistar rats (n=30). Defects were created on the proximal, the thickest part of the femur. The bone defects of the first group were filled with polymer/HA composite, the second group filled with only HA and the third group was left empty. Histologic examination of bone tissues showed new bone formation around the yellow-green polymer/HA composite material in the first group of animals whereas no evidence of new bone growth was observed in other groups.     

Inhibition of T-cell activation with HLA-DR1/DR4 restricted Non-T-cell stimulating peptides

It has been reported that collagen II (CII) derived peptide CII263-272 induced T-cell activation via its amino acids responsible for T-cell receptor (TCR) recognition. The impact of substitution of the TCR contacting amino acids of CII263-272 on T-cell activation was evaluated in this study using a panel of altered CII263-272 peptides. Computer modeling revealed that the side chains of 263F and 266E in CII263-272 were coupled with amino acids on alpha1 and beta1 chains of HLA-DR1 or -DR4, mainly via hydrogen bonds, whereas the side chains of 267Q and 270K protrude out of the cleft and might be recognized by TCR. Intracellular delivery of the altered peptides, and their binding to HLA-DR1 and -DR4 molecules on cell surface, were demonstrated by confocal microscopy and flow cytometry. The results also revealed that the substitution of 267Q, 268G, 269P, and 270K individually or consecutively by alanine (A) or glycine (G) led to weak or non-T-cell responses. Furthermore, the altered peptides with 270K substitution (270A) or with consecutive substitution of 268G, 269P, and 270K (sub268-270) dramatically inhibited T-cell activation. It is suggested that the altered peptides derived from CII263-272 with substitution of amino acids responsible for TCR contact might be of inhibitory effect on T-cell responses.     

Age-related alterations in responses of the nucleus basalis magnocellularis neurons to frontal cortex stimulation in rats

The present study investigated the age-related alterations in responses of the nucleus basalis magnocellularis (nbM) neurons to frontal cortex (FCX) stimulation. Single unit extracellular recording from the nbM neurons were obtained with glass micropipettes in urethane-anesthetized rats. A total of 137 units were located within the nbM in the three age groups (young, 3 months; adult, 12 months; old, 24 months). FCX stimulation elicited responses in 91% of the 137 neurons. Most of them were excited. The frequency of occurrence of excitatory responses in the nbM neurons was decreased with aging. The thresholds and latencies of excitatory responses evoked by FCX stimulation were increased in old rats. The mean peak-firing rate of exciting phase was gradually reduced with aging. These findings indicate that there might be some functional changes in the nbM neurons with aging.     

利用酵母双杂交技术筛选鉴定STCH与RanBP9的相互作用

以人应激和分子伴侣蛋白(STCH)为诱饵蛋白,利用酵母双杂交技术在高严格条件下筛选人脑cDNA文库。在SD/-4培养基上共获得156个阳性克隆,将酵母菌扩增后抽提质粒,转化大肠杆菌,进行序列分析,并经过回复杂交验证,潜在的与STCH相互作用的蛋白有:VDAC、MBP2、ZNF251、RanBP9、Phosophate glycolase、β-tubulin、up1、up2和WW adaptor蛋白。基于各候选蛋白与神经系统疾病的关系,选择RanBP9作进一步验证实验。经体外GST-Pulldown和体内免疫共沉淀实验证实RanBP9和STCH能够相互作用,明确RanBP9的羧基端为相互作用的最小功能域。此结果为我们进一步揭示STCH作用的分子机制提供了线索。     

特殊染色和免疫组织化学染色在肾上皮性肿瘤诊断中的应用

目的 探讨肾上皮性肿瘤的诊断及鉴别诊断。方法 对91例具有较完整病理资料的肾上皮性肿瘤进行了常规组织学观察,Mowy胶状铁染色及CD10、细胞角蛋白(CK)7、波形蛋白免疫组织化学染色。结果 91例肾上皮性肿瘤中肾透明细胞癌78例,占86％;肾乳头状癌8例,占9％;肾嫌色细胞癌4例,占4％;肾嗜酸细胞腺瘤1例,占1％。78例肾透明细胞癌中CD10、波形蛋白阳性分别为63例(81％)和69例(88％),主要表现为细胞膜阳性。74／78肾透明细胞癌CK7呈阴性,17／17 Mowy胶状铁染色阴性或呈灶状粗颗粒。肾嫌色细胞癌4例,CK7均呈胞膜阳性;Mowy胶状铁染色表现为胞质内蓝色细网状;4例CD10肿瘤细胞膜均阴性,波形蛋白阴性。肾嗜酸细胞腺癌1例CD10,CK7,波形蛋白,Mowy胶状铁染色均为阴性或不特异。结论 在常规诊断肾上皮性肿瘤有困难时免疫组织化学CD10,CK7,波形蛋白染色以及Mowy胶状铁染色对鉴别诊断有帮助。     

p73在骨巨细胞瘤中表达的意义

目的：研究p73蛋白在骨巨细胞瘤中的表达,探讨其在骨巨细胞瘤发生、发展中的可能作用。方法：应用免疫组化S－P法检测40例骨巨细胞瘤石蜡标本中p73蛋白的表达情况。结果：p73在骨巨细胞瘤中表达阳性率为30.0％,对照组骨软骨瘤表达阳性率为20.0％,两组相比无显著差异（P＞0.05）。p73在骨巨细胞瘤Ⅰ～Ⅱ级组表达阳性率为17.9％,Ⅲ级组为58.3％,二者相比差异显著（P＜0.05）。结论：p73在Ⅲ级骨巨细胞瘤中表达明显增高,提示其在骨巨细胞瘤发生、发展中可能发挥重要作用,并可作为骨巨细胞瘤Jaffe分级的辅助指标之一。