Prenatal molecular diagnosis of X‐linked hydrocephalus via a silent C924T mutation in the L1CAM gene

We present a case of a patient whose L1CAM gene in X‐chromosome has a C924T transition. Her first son's ventriculomegaly was prenatally detected. A mature infant was born, his head circumference was large, and thumbs were bilaterally adducted. X‐linked hydrocephalus (XLH) was suspected. The DNA examination revealed that both her and boy's LICAM gene had a C924T transition. She became pregnant 5 years later and amniocentesis was performed. The results of cytogenetic analysis revealed that the fetus was female. She continued her pregnancy and delivered a healthy girl. She again became pregnant 3 years later. The chromosomal analysis revealed that the fetus was male. Fetal DNA analysis determined that the fetus had the inherited mutation. She chose to terminate the pregnancy. A C924T mutation can be disease causing for XLH, and the detection of this mutation would aid in genetic counseling for the prenatal diagnosis of XLH.     

Analysis of YKL‐40 Acute‐Phase Protein and Interleukin‐6 Levels in Periodontal Disease

Background: YKL‐40, a new acute‐phase protein, is shown to be elevated in inflammatory diseases, such as rheumatoid arthritis, type 2 diabetes mellitus, and coronary artery diseases. However, there is no data indicating a relationship between YKL‐40 and periodontal disease. Interleukin‐6 (IL‐6) is the major regulator of acute‐phase protein synthesis and one of the most studied inflammatory markers in periodontal disease. The purpose of the present study is to evaluate YKL‐40 and IL‐6 levels in gingival crevicular fluid (GCF) and serum of patients with periodontal disease and healthy individuals. Methods: Periodontally healthy individuals (n = 15), patients with gingivitis (n = 15), and patients with severe chronic periodontitis (CP) (n = 15) without any systemic disease were included in the study. Clinical measurements were recorded; GCF and blood samples were obtained from each participant. GCF and serum YKL‐40 and IL‐6 levels were analyzed by enzyme‐linked immunosorbent assay. Statistical analysis was performed by parametric and non‐parametric tests. Results: Total amounts of YKL‐40 and IL‐6 in GCF as well as serum YKL‐40 and IL‐6 levels were significantly higher in patients with gingivitis and CP compared with healthy controls (P <0.01). YKL‐40 levels in GCF and serum as well as serum IL‐6 levels were significantly higher in patients with CP compared with patients with gingivitis (P <0.01). Conclusions: YKL‐40 levels in GCF as well as serum YKL‐40 and IL‐6 levels increased from gingivitis to periodontitis. Within the limits of the present study, the YKL‐40 molecule might be a potential novel inflammatory marker of periodontal disease.     

RNAi screening with shRNAs against histone methylation-related genes reveals determinants of sorafenib sensitivity in hepatocellular carcinoma cells

Sorafenib is the first drug currently approved to treat advanced hepatocellular carcinoma (HCC). However, very low response rate and acquired drug resistance makes rare patients benefit from sorafenib therapy, therefore it is urgent to find biomarkers for sorafenib sensitivity. Histone modifications, including histone methylation, have been demonstrated to influence the initiation and progression of HCC. It is of great interest to elicit the possibility whether histone methylation plays a role in regulation of sorafenib sensitivity. In present work, a high throughput RNAi screening with 176 shRNA pools against 88 histone methyltransferases (HMTs) and histone demethyltransferases genes was applied to HepG2 cells. Silencing of 3 genes (ASH1L, C17ORF49 and SETD4) was validated to specifically promote HepG2 cells sensitivity to sorafenib. Western blotting results showed that those 3 HMT genes knockdown alone or sorafenib treatments alone both induce AKT/ERK activation. However, combination treatment with sorafenib and silencing of C17ORF49 or SETD4 downregulated AKT phosphorylation and hence induced HCC cells death. Our work may provide potential biomarkers for sorafenib sensitivity and therapeutic combination for sorafenib treatment in HCC patients.     

CD40L基因c.674T>C突变的X连锁高IgM综合征1例报道

目的探讨高IgM综合征的特点、临床表现、实验室检查与治疗。方法回顾性分析1例X连锁高IgM综合征患者的临床资料。结果患儿以反复呼吸道感染入院,免疫功能检查提示IgG、IgA明显降低,IgM明显升高;基因检测结果示CD40L基因c.674T>C突变,给予抗感染、定期静脉滴注人免疫球蛋白等治疗。结论高IgM综合征临床表现缺乏特异性,依靠基因检测进行诊断;规律人免疫球蛋白替代治疗可能会提高患儿生存质量,延长寿命。     

The clinical spectrum of mutations in L1, a neuronal cell adhesion molecule

Mutations in the gene encoding the neuronal cell adhesion molecule L1 are responsible for several syndromes with clinical overlap, including X-linked hydrocephalus (XLH, HSAS), MASA (mental retardation, aphasia, shuffling gait, adducted thumbs) syndrome, complicated X-linked spastic paraplegia (SP 1), X-linked mental retardation-clasped thumb (MR-CT) syndrome, and some forms of X-linked agenesis of the corpus callosum (ACC). We review 34 L1 mutations in patients with these phenotypes. © 1996 Wiley-Liss, Inc.     

Reduction of hippocampal long-term potentiation in transgenic mice ectopically expressing the neural cell adhesion molecule L1 in astrocytes

The influence of the neural cell adhesion molecule L1 on hippocampal long-term potentiation (LTP) was investigated using transgenic mice ectopically expressing L1 in astrocytes (GFAP-L1). L1 is a member of the immunoglobulin superfamily of homophilic adhesion molecules predominantly expressed in neurones. Previously, it has been demonstrated that local application of L1 antibodies and recombinant L1 fragments impair the expression of LTP. Here, we show that LTP induced by theta-burst stimulation or by pairing presynaptic stimulation with postsynaptic depolarisation was strongly reduced in GFAP-L1 mice, whereas basal synaptic transmission, post-tetanic potentiation, and paired-pulse facilitation were not modified. These results further support the idea that L1 is involved in synaptic plasticity and suggest that adhesion molecule-dependent changes in synaptic morphology contribute to the expression of LTP. © 1996 Wiley-Liss, Inc.     

红花SRAP扩增体系的建立和优化

目的:探讨影响红花SRAP扩增的各种因素,建立能够稳定扩增红花基因组的体系,为研究红花重要性状的遗传基础及建立分子辅助标记育种的技术平台奠定基础.方法:用CTAB法提取红花DNA,设计Taq酶浓度(0.02、0.04、0.06 U/μl)、dNTP浓度(0.15、0.25、0.30 mmol/L)、Primer浓度(0.15、0.30、0.45 μmol/L)3因素3水平27次实验和Mg2 浓度(0.5、1.0、1.5、2.0、2.5、3.0、3.5、4.0 mmol/L)8水平单因素实验.在25 μl体系中加入模板DNA 20 ng.对体系进行优化,用琼脂糖进行检测.结果:本研究建立了适合红花的SRAP体系,Taq酶浓度为0.02 U/μl,dNTP浓度为0.25 mmol/L,Primer 浓度为0.30 μmol/L,Mg2 的浓度为3.0 mmol/L,优化后的体系目标条带增多,重现性好,得到了较好的扩增效果.结论:本研究建立的反应体系适合红花SRAP的研究.     

GITR抗体对L615白血病抑制作用的实验研究

目的 探讨糖皮质激素诱发型TNF受体(glucocorticoid-induced tumor necrosis factor receptor,GITR)的抗体对抗小鼠L615白血病(T淋巴细胞来源白血病)的效果和作用机制.方法 以L615小鼠建立白血病模型,分3组实验组以及阴性对照组,观察实验小鼠生存状态及时间、外周血白细胞计数及分类、外周血和骨髓中白血病细胞形态变化、肝脾指数、肝脾组织病理改变.结果 GITR抗体可以延长L615白血病小鼠的生存时间,可引起骨髓中白血病细胞发生凋亡、坏死、肝脾指数降低、肝脾组织被白血病细胞浸润,提示GITR抗体能够降低和缓解白血病细胞所致的小鼠外周血白细胞的升高和浸润,以及肝脾的肿大.结论 通过免疫调节机制GITR抗体能够有效地抑制L615白血病细胞的增殖,进而抑制白血病的发展.     

合成紫花茄皂甙诱导人肝癌细胞株Bel-7402凋亡的作用机制

背景与目的:紫花茄(SolanumindicumL.)是一种有抗炎和治疗跌打损伤作用的传统中草药,含有丰富的结构独特的薯蓣皂甙,即紫花茄皂甙。我们的研究结果已证实,数种合成紫花茄皂甙有强抗肿瘤效果。本研究探讨合成紫花茄皂甙Ⅰ("-D-木吡喃糖基-(1→3)-[#-L-鼠李吡喃糖基-(1→2)]-"-D-半乳吡喃糖基薯蓣苷)的抗肿瘤机制。方法:以不同浓度紫花茄皂甙处理人肝癌细胞Bel-7402后,用酸性磷酸酶(acidphosphataseassay,APA)法测定增殖抑制率和IC50。用结晶紫染色显微镜观察细胞的形态学变化。用蛋白免疫印迹法检测凋亡过程中相关蛋白的变化。结果:紫花茄皂甙对Bel-7402细胞有明显的增殖抑制作用,在皂甙Ⅰ作用72h后的IC50为4.20μg/ml。光学显微镜下可见细胞密度降低,胞质中出现膜泡等形态学变化。蛋白免疫印迹检测结果显示,紫花茄皂甙Ⅰ作用后细胞质中的细胞色素c含量明显增加,Caspase-3被激活,细胞内的poly(ADR-ribose)polymerase(PARP)蛋白被剪切。结论:紫花茄皂甙对人肝癌细胞的增殖抑制呈浓度相关性,且通过线粒体依赖性通路诱导肿瘤细胞凋亡。