Molecular and biological analysis of echovirus 9 strain isolated from a diabetic child

The full-length infectious cDNA clone was constructed and sequenced from the strain DM of echovirus 9, which was recently isolated from a 6-week-old child at the clinical onset of type 1 diabetes. Parallel with the isolate DM, the full-length infectious cDNA clone of the prototype strain echovirus 9 Barty (Barty-INF), was constructed and sequenced. Genetic relationships of the sequenced echo 9 viruses to the other members of the human enterovirus type B species were studied by phylogenetic analyses. Comparison of capsid protein sequences showed that the isolate DM was closely related to both prototype strains: Hill and Barty-INF. The only exception was the inner capsid protein VP4 where serotype specificity was not evident and the isolate DM clustered with the strain Hill and the strain Barty-INF with echovirus 30 Bastianni. Likewise, the nonstructural protein coding region, P2P3, of isolate DM was more similar to strain Hill than to strain Barty-INF. However, like echovirus 9 Barty, the isolate DM contained the RGD-motif in the carboxy terminus of capsid protein VP1. By blocking experiments using an RGD-containing peptide and a polyclonal rabbit antiserum to the alpha(v)beta(3)-integrin, it was shown that this molecule works as a cellular receptor for isolate DM. By using primary human islets, it was shown that the isolate DM is capable of infecting insulin-producing beta-cells like the corresponding prototype strains did. However, only isolate DM was clearly cytolytic for beta-cells. The infectious clones that were made allow further investigations of the molecular features responsible for the diabetogenicity of the isolate DM.     

Direct molecular diagnosis of myotonic dystrophy

Hecht BK, Donnelly A, Gedeon AK, Byard RW, Haan EA, Mulley JC. Direct molecular diagnosis of myotonic dystrophy. Clin Genet 1993: 43: 276–285. © Munksgaard, 1993 Myotonic dystrophy (DM) arises from an unstable trinucleotide (CTG_n) repeat sequence within the DM locus at 19q13.3. Twenty-three myotonic dystrophy families containing 205 persons with no symptoms, minimal manifestations, classic DM or congenital DM were investigated to validate the application of the pM10M6 probe to direct molecular diagnosis. Affected family members had been diagnosed clinically and the unaffected family members had been assigned carrier probabilities close to either zero or 100%, using closely linked flanking markers. Southern analysis identified all 89 DM gene carriers as having expansions of the unstable element. PstI detected all small expansions of the repeat sequence as easily seen discrete bands; but large expansions were usually seen as diffuse smears, sometimes difficult to distinguish from lane background. EcoRI concentrated these diffuse smears, associated with somatic instability, into discrete bands which were easy to detect; but it did not resolve the smaller expansions present in 9 (10%) of the DM carriers. It is essential that PstI and EcoRI gels are run in parallel to detect all DM gene carriers. The extent of expansion of CTG correlated with age of onset and disease severity. Biopsies of various fetal tissues from two terminated pregnancies confirmed the diagnosis obtained by CVS and revealed no heterogeneity between tissues at this developmental stage. Further expansion occurred during the culture of CVS cells, indicating that direct prenatal diagnosis needs to be carried out on CVS tissue rather than on cultured cells. The intergenerational change of the repeat sequence from DM parent to DM offspring showed a significant parental sex difference for those parents with large expansions. Contraction of the unstable element was observed in the three males carrying the largest expansions and could explain why congenital DM is exclusively of maternal origin.     

Genetic variation screening and association studies of the adenylate cyclase activating polypeptide 1 (ADCYAP1) gene in patients with type 2 diabetes

Adenylate cyclase activating polypeptide 1 (ADCYAP1) is a pancreatic neuropeptide and modulates glucose-stimulated insulin secretion. The ADCYAP1 gene is located on chromosome 18p11 linked to type 2 diabetes. To test whether it is a candidate gene for type 2 diabetes, screening of the gene in Finnish and Swedish type 2 diabetic patients was done. Two novel SNPs, g.9863G>A (G54D) in exon 3 and g.12712C>G in the 3'-UTR of exon 5 of the ADCYAP1 gene (accession number X60435), were found. PCR-RFLP genotyping was then performed in a total of 253 type 2 diabetic patients and 253 non-diabetic control subjects. Transmission disequilibrium test (TDT) was performed in 132 parent-offspring trios. The G allele frequencies of g.9863G>A (G54D) and g.12712C>G of the ADCYAP1 gene were higher in type 2 diabetic patients than in non-diabetic control subjects (21.0% vs 15.8%, P=0.04; 5.3% vs 3.0%, P=0.045). However, no significant differences in clinical variables was seen between the different genotype carriers, and also no transmission distortion of the G allele of SNP g.9863G>A (G54D) was observed in 132 parent-offspring trios. The present study thus suggest that the variants in the ADCYAP1 gene may not be major influence of the susceptibility to type 2 diabetes in Finnish and Swedish Caucasians.     

Analysis of children with type 1 diabetes in Korea: high prevalence of specific anti-islet autoantibodies, immunogenetic similarities to Western populations with "unique" haplotypes, and lack of discrimination by aspartic acid at position 57 of DQB

This study analyzed the expression of anti-islet autoantibodies and HLA-DR and -DQ genotypes in Korean children with type 1 diabetes mellitus (T1DM). The positivity of the anti-ICA512, anti-GAD65, and anti-insulin autoantibodies in the newly onset T1DM patients (n = 15) was 66.7%, 86.7%, and 46.7%, respectively, and all of them had one or more of the autoantibodies. HLA analysis showed higher frequencies of HLA-DRB1*0301, *0405, *09012 and -DQB1*0201, *0401, *03032 alleles in T1DM patients compared to controls (P(c) < 0.05). Because HLA-DQB1*0401, *03032 alleles carry aspartic acid at position 57 of DQB, susceptibility to T1DM in Korean children was not related to the presence of aspartic acid at position 57 of DQB1 locus. We suggest this unique HLA-DR, -DQ allele distribution might be an important factor for the low incidence of T1DM in Korea, and the combined anti-islet autoantibody assays could be valuable screening markers for the early detection of T1DM in Korea.     

091 HLA-DM及HLA-DO的研究进展

抗原递呈涉及多种机制及各种分子.细胞膜上的外来抗原被细胞表面的mIg或BCR摄取进入细胞内早/后期内涵体及溶酶体中.内质网中的新合成并结合于Ii链的HLAⅡ类分子进入内涵体/溶酶体中,Ii链降解成为CLIp片段,在DM/DO的联合作用下,结合于HLAⅡ类分子抗原结合槽内的CLIP脱落,同时抗原肽进入槽内形成HLAⅡ-抗原肽复合物,将抗原提呈于细胞表面供T细胞识别,激发一系列的免疫反应.DM/DO对于这一复杂过程的准确进行起着至关重要的作用.     

C-reactive protein concentration is not related to islet autoimmunity status in offspring of parents with type 1 diabetes

Autoimmunity may be associated with acute or chronic inflammation. In order to determine whether the inflammatory marker C-reactive protein (CRP) was an indicator of inflammatory events that precede, predict, or associate with islet autoimmunity or type 1 diabetes, CRP was measured in sequential antibody-negative, seroconversion, and follow-up-positive samples from 65 prospectively studied islet autoantibody-positive children. Although changes in CRP concentrations were observed in some children, overall CRP concentrations were similar in antibody-negative samples (median, 0.21 mg/L), antibody-positive samples (median, 0.26 mg/L), and samples at seroconversion (median, 0.26 mg/L). CRP concentrations at diabetes onset (median, 0.59 mg/L) were not significantly increased over antibody-negative samples (P = 0.07). CRP concentrations did not predict diabetes development. CRP concentrations were related to age (r = 0.26; P < 0.001) and were increased in samples obtained from October to January (P < 0.001). These findings suggest that CRP concentrations are not a valuable marker of progression to type 1 diabetes and highlight the importance of correcting analyses for seasonal variations.     

金芪降糖片联合西药治疗糖尿病对患者血浆ET-1和sICAM-1水平的影响

目的：探讨金芪降糖片对糖尿病患者血浆ET-1和sICAM-1水平的影响及临床应用价值。方法：金芪降糖片和降糖类西药同时服用治疗糖尿病，并与只服西药治疗组进行比较。采用酶联免疫法检测两组患者治疗前后血浆中血管内皮细胞活性因子ET-1和sICAM-1水平，同时观察血糖和血脂、HbA1c等各项指标的变化。结果：金芪组治疗后血糖和血脂、HbA1c均有显著变化（P〈0．05），对照组仅血糖有明显变化（P〈0．05）；金芪组治疗后ET-1和sICAM-1均显著降低，分别为P〈0．05和P〈0．01，对照组两者水平皆无明显变化（P〉0．05）。结论：金芪降糖片通过降低糖尿病患者血浆中ET-1和s／CAM-1水平，从而保护患者的血管内皮细胞，在西药常规治疗糖尿病的基础上，再加以中药辨证施治，可以提高临床对糖尿病的疗效。     

消渴灵冲剂治疗糖尿病的实验研究

目的 :观察消渴灵冲剂对Alloxan糖尿病大鼠的治疗作用。方法 :设立高、低剂量组与西药二甲双胍组作对照 ,动态地观察大鼠FBG、IRI及血流变等指标。结果 :消渴灵可降低Alloxan糖尿病大鼠异常升高的FBG、GHb水平 ,升高空腹IRI水平 ,与治疗前相比差异显著 (P <0 .0 1)。其降糖作用与二甲双胍组相比无差异。消渴灵可降低Alloxan糖尿病大鼠异常升高的Hr、Lr、Br等血流变指标 ,对抗血液的高凝高粘状态 ,其疗效优于二甲双胍 (P <0 .0 1) ,消渴灵有助于糖尿病大鼠恢复体重。结论 :其机理可能在于促进胰岛 β细胞的分泌功能 ,升高血中IRI水平 ,增进细胞对葡萄糖的利用 ,增强免疫功能 ,改善营养状态 ,改善血流变等。     

深圳市居民体质指数,腰臀围比与糖尿病的关系研究

采用分层整群抽样方法,研究了深圳市５ 个行政区年龄在２０ 岁以上,在深圳居住５ 年及以上８２００ 名常住社区居民肥胖与糖尿病的关系结果表明,深圳居民体质指数（ＢＭＩ） ≥２５ 的人群糖尿病患病率和ＩＧＴ 患病率明显高于体质指数（ＢＭＩ） ＜２５ 的人群,肥胖人群糖尿病患病率和ＩＧＴ 患病率分别是非肥胖人群的２．６ 倍和２３ 倍。随腰围、臀围及两者之比的升高,糖尿病患病率和ＩＧＴ患病率升高;腰臀围比≥１ 和＜１人群糖尿病患病率和ＩＧＴ 率之比分别为１：５ 和１：２５。研究结果还表明,随腰围的增加两率增加更加明显,腰围平均每增加１ｃｍ 糖尿病患病率增加０５％ ,ＩＧＴ 患病率增加１０％     

应用胰岛素泵治疗儿童及青少年1型糖尿病对糖代谢的影响

为观察应用胰岛素泵治疗儿童及青少年1型糖尿病(T1DM)对糖代谢的影响 ,随访10例胰岛素泵治疗的T1DM患儿 ,分别观察胰岛素泵治疗前、后6个月的糖化血红蛋白值(HbA1c)、胰岛素用量、严重低血糖及酮症酸中毒发生次数的变化情况。结果显示 ,胰岛素泵治疗6个月后HbA1c 显著下降 ,治疗前为8.97 %±1.69 %,治疗后为7.51 %±1.17 % (t=2.52 ,P<0.05) ;胰岛素用量无显著下降 ;未发生严重低血糖和酮症酸中毒。表明胰岛素泵治疗可有效控制血糖 ,明显降低HbA1c,减少低血糖及酮症酸中毒的发生 ,是儿童及青少年T1DM常规治疗的较好选择。