ICU重症IgA肾病患者IL-4、C1GALT1及COSMC表达对疾病预后的调控作用分析

目的 探讨在重症IgA肾病患者中IL-4、C1GALT1及COSMC的mRNA表达及潜在预后因素对疾病及其预后的调控作用.方法 选取本院ICU的重症确诊IgA患者61名,同时选取年龄性别可比的体检健康者65名作为对照,并使用荧光定量PCR测定外周血的单核细胞IL-4、C1GALT1以及COSMC的mRNA表达,同时问卷收集可能影响预后的相关因素,IgA患者随访5年,以随访结束时出现终末期肾病为预后不佳进行研究分析.对相关因素对本病预后的调控作用进行分析讨论.患者均知情同意并签署知情同意书.统计分析采用卡方检验以及t检验对一般资料及两组基因表达进行比较,使用多因素Logistics回归模型进行预后因素讨论,使用SPSS17.0统计软件包进行数据分析.结果 IgA患者组IL-4mRNA的表达水平为3.67±0.34,显著高于对照组的1.62±0.16;IgA组的C1GALT1 mRNA以及COSMC mRNA表达水平分别为1.42±0.22和1.34 ±0.13,出现相应的表达降低;对照组分别为1.61 ±0.22和1.63±0.18.对ICU重症IgA肾病患者进行预后分析,单因素分析结果提示IL-4表达增加在预后不良者中的构成显著高于预后良好者,P＜0.05,肾病家族史构成比例在预后不良者中也明显较高.多因素Logistic回归结果提示年龄大于60岁、肌酐超标、高血压、肾病家族史以及IL-4 mRNA表达增加和C1GALT1 mRNA表达减低均为IgA肾病不良预后的危险因素.结论 ICU重症IgA肾病患者IL-4 mRNA表达增加,同时可见相应的C1GALT1 mRNA以及COSMC mRNA表达减低,且这三个因素与高龄、肌酐异常、高血压等因素同为重症IgA的预后不良因素.     

双价志贺菌苗灌胃免疫小鼠后肠粘膜的免疫应答

为了观察两株福氏、宋内双价志贺菌苗 (Ipa ,Ipa+ )免疫后所引起的肠粘膜诱导部位的免疫应答及为研制有效腹泻疫苗提供理论基础。应用小鼠灌胃免疫为模型 ,以间接免疫荧光法和BA ELISPOT方法 ,检测了肠粘膜相关淋巴组织 (gutasso ciatedlymphoidtissues ,GALT )中诱导部位派伊尔小结 (Payer’spatches ,PP )、肠系膜淋巴结 (mesentericlymphoidnode ,MLN )T淋巴细胞亚群和特异性抗体分泌细胞的变化 (ASC )及小肠局部和系统的特异性抗体变化。发现两株双价菌苗株均可引起肠粘膜GALT诱导部位CD4+ 淋巴细胞亚群、抗体分泌细胞明显升高 ;同时局部及血清中特异性抗体明显升高。说明双价志贺菌苗灌胃免疫小鼠后 ,诱导部位的免疫应答早期以体液免疫为主 ,灌胃不仅可诱导局部特异性抗体免疫应答也可诱导系统的抗体免疫应答。     

The ability of an LC-MS/MS-based erythrocyte GALT enzyme assay to predict the phenotype in subjects with GALT deficiency

BackgroundGALT deficiency is a rare genetic disorder of carbohydrate metabolism. Due to the decreased activity or absence of the enzyme galactose-1-phosphate uridylyltransferase (GALT), cells from affected individuals are unable to metabolize galactose normally. Lactose consumption in the newborn period could potentially lead to a lethal disease process with multi-organ involvement. In contrast to the newborn-stage disease, however, a galactose-restricted diet does not prevent long-term complications such as central nervous system (CNS) dysfunction with speech defects, learning disability and neurological disease in addition to hypergonadotropic hypogonadism or primary ovarian insufficiency (POI) in females. As the literature suggests an association between GALT enzyme activity and the long-term complications, it is of importance to have a highly sensitive assay to quantify the GALT enzyme activity. To that end, we had developed a sensitive and accurate LC-MS/MS method to measure GALT enzyme activity. Its ability to predict outcome is the subject of this report.Materials and methodsThe GALT enzyme activity in erythrocytes from 160 individuals, in which 135 with classic, clinical variant or biochemical variant galactosemia, was quantified by LC-MS/MS. Individuals with GALT deficiency were evaluated for the long-term complications of speech defects, dysarthria, ataxia, dystonia, tremor, POI, as well as intellectual functioning (full scale IQ). The LC-MS/MS results were compared to a variety of assays: radioactive, [¹⁴C]-galactose-1-phosphate, paper chromatography with scintillation counting, enzyme-coupled assays with spectrophotometric or fluorometric readout or high-pressure liquid chromatography with UV detection of UDP-galactose.ResultsThe LC-MS/MS method measured GALT activity as low as 0.2%, whereas other methods showed no detectable activity. Largely due to GALT activities that were over 1%, the LC-MS/MS measurements were not significantly different than values obtained in other laboratories using other methodologies. Severe long-term complications were less frequently noted in subjects with >1% activity. Patients with a p.Q188R/p.Q188R genotype have no residual enzyme activity in erythrocytes.ConclusionOur LC-MS/MS assay may be necessary to accurately quantify residual GALT activities below 5%. The data suggest that patients with >1% residual activity are less likely to develop diet-independent long-term complications. However, much larger sample sizes are needed to properly assess the clinical phenotype in patients with residual enzyme activities between 0.1 and 5%.     

Involvement of peripheral and central catecholamine systems in neural-immune interactions

In this review, we have attempted to delineate the current state of knowledge of the relationships between the immune system and one chemically specific component of the nervous system, the noradrenergic system, both in the brain and the periphery. We have discussed recent work describing the presence of noradrenergic innervation in lymphoid tissues in the major lymphatic organs. Our findings demonstrate clearly that the regions in which lymphocytes (mainly T cells) reside, and through which they recirculate, receive direct sympathetic neural input. The immune system can, therefore, be considered 'hard-wired' to the brain. The evidence for receptors on cells of the immune system capable of receiving signals from the brain is discussed. The significance of this 'hard-wiring' to the function of the immune system is considered, both with regard to the effect of its disruption on immune responses, and to the direct and indirect effects of sympathetic neurotransmitter substances on lymphocytes and their behavior in vitro and in vivo. Finally, our detailed analysis of changes occurring in central noradrenergic pathways as a result of stimulation of the immune system leads to an emerging picture of feedback loops from the immune system to the brain. Such circuits employ endocrine, and probably autonomic, outflow to modulate and regulate immune responses.     

Identification of mutations in the galactose‐1‐phosphate uridyltransferase (GALT) gene in 16 Turkish patients with galactosemia,including a novel mutation of F294Y

Classical galactosemia caused by deficiency of galactose‐1‐phosphate uridyltransferase (GALT) is a severe autosomal recessive disorder. We report here molecular analysis of 16 unrelated Turkish galactosemia index cases without GALT activity. Almost 84% of all mutant alleles were identified in this study. The most common molecular defect observed in the Turkish population was Q188R (replacement of glutamine‐188 by arginine) (57%). In order to facilitate the determination of unknown mutations in the entire coding region of GALT, we established an approach based on GALT cDNA synthesis and direct sequencing. We have identified one novel candidate galactosemia mutation, a T‐to‐A transversion at the codon 294 (F294Y) in exon 9 in addition to previously reported three missense (M142K K285N, A320T), one stop codon (E340X), and one silent (L218L) mutations in galactosemia patients which reflect considerable genetic heterogeneity in the Turkish population. © 1999 Wiley‐Liss, Inc.     

A one-year oral toxicity study of sodium stearoyl lactylate (SSL) in rats

The toxicity of sodium stearoyl lactylate (SSL) was examined in Wistar rats fed diets containing 0, 1.25, 2.5, and 5% SSL for one year, equivalent to mean daily intakes of 558, 1115, and 2214 mg/kg/day in males and 670, 1339, and 2641 mg/kg/day in females, respectively. SSL was well tolerated at these dietary levels as evidenced by the absence of toxicologically significant changes in the general condition and appearance of the rats, survival, neurobehavioral endpoints, growth, feed and water intake, ophthalmoscopic examinations, hematology and clinical chemistry parameters, urinalysis, or necropsy findings. The occurrence of uterine endometrial stromal polyps was the only finding of potential significance. Given the frequent occurrence of these benign tumors in rats, wide variability in the reported incidence of this type of polyps in rats, the lack of statistical significance and lack of biological evidence to suggest a mechanism for the slightly greater incidence in the groups fed 2.5 and 5% SSL, it was concluded that the endometrial stromal polyps observed in females fed SSL were not related to treatment. The no observed adverse effect level (NOAEL) of SSL was placed at 5%, the highest dietary level tested (equivalent to 2214 mg/kg/day for males and 2641 mg/kg/day for females).     

Up-regulation of C1GALT1 promotes breast cancer cell growth through MUC1-C signaling pathway

Aberrant glycosylation is frequently observed in cancers. Core 1 β1,3-galactosyltransferase (C1GALT1) is an exclusive enzyme in humans that catalyzes the biosynthesis of core 1 O-glycan structure, Gal-GalNAc-O-Ser/Thr, whose expression is commonly up-regulated during tumorigenesis. Little is known about the function of C1GALT1 in breast cancer. This study aims to determine the correlation between C1GALT1 expression and breast cancer clinicopathological features and roles of C1GALT1 in breast cancer malignant phenotypes. Public databases and our data showed that C1GALT1 mRNA and C1GALT1 protein are frequently up-regulated in breast cancer; and increased C1GALT1 expression correlates with higher histological grade and advanced tumor stage. Overexpression of C1GALT1 enhanced breast cancer cell growth, migration, and invasion in vitro as well as tumor growth in vivo. Conversely, C1GALT1 knockdown suppressed these malignant phenotypes. Furthermore, C1GALT1 modulates O-glycan structures on Mucin (MUC) 1 and promotes MUC1-C/β-catenin signaling in breast cancer cells. These findings suggest that C1GALT1 enhances breast cancer malignant progression through promoting MUC1-C/β-catenin signaling pathway. Unveiling the function of C1GALT1 in breast cancer opens new insights to the roles of C1GALT1 and O-glycosylation in tumorigenesis and renders the potential of C1GALT1 as a target of novel therapeutic agent development.