Immunoglobulin variable gene analysis of human autoantibodies reveals antigen-driven immune response to glutamate decarboxylase in type 1 diabetes mellitus

Insulin-dependent (type 1) diabetes mellitus is an organ-specific autoimmune disease frequently associated with an islet-specific humoral autoimmune response. The role of islet cell autoantibodies in the disease process is unclear; in particular, it is not known whether they are a non-specific side effect of islet cell destruction or play a role in the autoimmune network leading to type 1 diabetes. Here we report the immunoglobulin gene usage and somatic mutation rates of a panel of seven human monoclonal islet cell autoantibodies (MICA 1–7) directed towards the major islet cell autoantigen glutamate decarboxylase (GAD). These autoantibodies were produced from cells from two patients with newly diagnosed type 1 diabetes. VH1, VH4 and Vλ2 gene segments were frequently used in the MICA, but no correlation between V gene usage and epitope recognition was found. The nonrandom ratio of replacement versus silent mutations in the variable gene region, an accumulation of replacement mutations in the complementarity determining regions, which confer antigen binding, and the high relative avidity for GAD observed for MICA 1, 3, 4, and 6, suggested that the immune response to GAD is driven by the antigen. In contrast, MICA 2, 5, and 7, revealing a lower affinity for antigen, have accumulated a large number of silent mutations. These latter antibodies may, therefore, be characteristic for later stages of the chronic autoimmune disease. Our results argue in favor of an antigen-driven autoantibody response to islets in human type 1 diabetes. They suggest that GAD is an important target of autoimmunity associated with type 1 diabetes.     

基于中医临床数据的潜变量建模分析

目的结构方程模型(structural equation model,SEM)是一种建立、估计和检验因果关系的潜变量建模方法,目前在心理、教育和社会科学等学科领域中得到广泛的应用。中医临床数据中蕴含着许多重要但无法直接测量的潜变量(如证候),传统的一阶验证性因子分析方法仅能够分析单层次潜变量与症状指标间的相关性。本文提出基于中医临床数据的二阶验证性因子分析潜变量建模方法,对两个层次的证候及其症状指标进行相关性分析,从而为医生进一步定量理解疾病进展规律提供一定帮助。方法针对非小细胞肺癌中医临床数据特点,将中医证候和虚实辨证证候作为两阶因子,对其与症状指标进行潜变量建模分析。结果实验结果验证了非小细胞肺癌临床数据中的中医证候与虚实辨证证候及其与不同症状指标的相关关系。结论中医证候与虚实辨证证候以及症状指标间存在一定的内在相关规律,本研究对中医证候的规范化影响具有一定的意义。     

慢性淋巴细胞白血病患者免疫球蛋白重链片段及其突变状态的研究

目的 探讨我国慢性淋巴细胞白血病(CLL)患者免疫球蛋白重链可变区基因(IgVH)突变状态、基因偏颇性及其预后价值.方法 应用多重PCR法检测CLL患者表达型IgVH片段使用情况及突变状态,并与患者生存和预后进行相关性分析.结果 ①共检测初诊CLL患者85例,80例(94.1%)成功测序.②IgVH片段的使用存在明显的偏颇性,其中常见的为VH3家族(32例,40.0%)、VH4家族(24例,30.O%),而VH5、VH6和VH7家族少见.VH4-39是最常见的VH基因片段.而J4和D3分别是JH家族和DH家族中使用频率最高的片段.③IgVH突变型56例(70.0%),非突变型24例(30.O%).其中VH3家族突变率为90.7%(29例),VH4家族突变率为62.5%(15例).④突变组与未突变组患者无进展生存时间(82个月和17个月,P=0.000)和总生存时间(90个月和41个月,P=0.009)差异均有统计学意义.⑤CLL患者IgVH CDR3区序列具重现性,2例VH片段为VH1-69的患者其CDR3序列与文献报道序列有很高的同源性.VH片段为VH1-18、VH3-7、VH3-23的患者中分别有2例CDR3序列完全相同.结论 不同地区间CLL患者IgVH基因家族使用情况和突变状态存在一定差异,IgVH CDR3区序列具重现性,推测相对有限的抗原参与了CLL的发病.     

蛋白转导结构域介导乙型肝炎病毒聚合酶末端蛋白重链可变区抗体体外抑制病毒的复制

目的 蛋白转导结构域(TAT)介导HBV聚合酶末端蛋白(TP)重链可变区(VH)抗体,研究特异性TAT-VH抗体体外对HBV复制的影响.方法 将TAT-VH基因克隆入原核表达载体pET28a(+),在大肠埃希菌BL21(DE3)LysS内诱导融合蛋白表达并进行纯化.纯化的TAT-VH加入培养的HepG2.2.15细胞,间接免疫荧光法检测其导入HepG2.2.15细胞的效率,四甲基偶氮唑盐(MTT)法检测其对细胞生长代谢的影响,将TAT-VH加入培养的HepG2.2.15细胞,定量PCR法检测HBV DNA水平.数据行单因素方差分析和t检验.结果 成功制备了TAT-VH融合蛋白,间接免疫荧光及MTT证实TAT-VH可以跨膜导入HepG2.2.15细胞,且对细胞生长无影响;加入5 000 nmol/L TAT-VH的HepG2.2.15细胞培养上清液内HBV DNA为(1.211±0.132)lg拷贝/mL,对照组为(5.325±0.041)lg拷贝/mL(t=72.91,P＜0.05);细胞内分别为(3.521±0.411)和(8.532±0.132)Ig拷贝/mL(t=28.41,P＜0.05).结论 HBV聚合酶TP区特异性TAT-VH抗体在体外可抑制HBV复制,为应用细胞内抗体治疗HBV感染提供了良好的实验基础.     

Feasibility of open and percutaneous corrective osteotomies of the second phalanx of the great toe: An approach on a cadaveric model

BackgroundHaving had a previous experience of 4 open F2 osteotomies to correct interphalangeal hallux valgus, the aim of this study was to test the efficacy of a percutaneous approach for this correction.Methods3 open and 12 percutaneous F2 osteotomies in 15 cadaver feet were performed. Interphalangeal (IPH), F2 asymmetry (AF2) and joint deviation (JD) angles were measured on radiographs. The operated great toes were anatomically analyzed looking for possible lesions to surrounding tissues.ResultsMean IPH decreased from 10.7° to 2.9°, AF2 from 7.8° to 1°, and JD from 1.4° to 0.5°. Damage to the hallux extensor tendon was detected in 1 foot and a nail bed lesion was detected in 1 case, both operated percutaneously.ConclusionsF2 Valgus deformity can be corrected using open or percutaneous osteotomy. The authors consider the open correction worthwhile because percutaneous techniques may damage surrounding tissues and the incision length difference is minor.Level of clinical evidence: 3.     

Penalized variable selection for accelerated failure time models with random effects

Accelerated failure time (AFT) models allowing for random effects are linear mixed models under the log-transformation of survival time with censoring and describe dependence in correlated survival data. It is well known that the AFT models are useful alternatives to frailty models. To the best of our knowledge, however, there is no literature on variable selection methods for such AFT models. In this paper, we propose a simple but unified variable-selection procedure of fixed effects in the AFT random-effect models using penalized h-likelihood (HL). We consider four penalty functions (ie, least absolute shrinkage and selection operator (LASSO), adaptive LASSO, smoothly clipped absolute deviation (SCAD), and HL). We show that the proposed method can be easily implemented via a slight modification to existing h-likelihood estimation procedures. We thus demonstrate that the proposed method can also be easily extended to AFT models with multilevel (or nested) structures. Simulation studies also show that the procedure using the adaptive LASSO, SCAD, or HL penalty performs well. In particular, we find via the simulation results that the variable selection method with HL penalty provides a higher probability of choosing the true model than other three methods. The usefulness of the new method is illustrated using two actual datasets from multicenter clinical trials.     

Using online content uniformity measurements for rapid automated process development exemplified via an X-ray system

This paper addresses the relevance of automated content testing for the rapid automated process development (RAPD). Our previous work demonstrated that RAPD allowed a fast and efficient development of a continuous capsule-filling process. Target was the mean weight and the relative standard deviation of the weight. Likewise important are the content and the content uniformity. However, an implementation demands a certain level of automation. In general, technology is available that can detect active pharmaceutical ingredient (API) inside the capsules but the final application is linked to additional development and investment in machinery. To eliminate doubts regarding the benefits of an automated content check within the RAPD we present an application example. First, an X-ray system was used to detect barium sulfate accurately inside capsules. Second, a process was developed where barium sulfate was filled. The concentration of excipients was modified in the experiments, as well as the setting of the process parameter. The obtained model provided an explicit understanding of the process. Subsequently, the content uniformity model was compared to a model of the capsule weight relative standard deviation, confirming the benefits of an automated content check in the RAPD. Moreover, we presented another example illustrating the advantages of a connected continuous filling process, which permits evaluation of all process steps and their interactions (i.e. evaluation of the entire process).     

Therapeutic plasma exchange for exogenous insulin antibody syndrome in combined variable immunodeficiency: a case report

A 32-year-old male with type I diabetes presented with profound hypoglycemia due to exogenous insulin antibody syndrome in the setting of newly-diagnosed common variable immunodeficiency. Immunomodulatory therapy was not initially effective, but after the initiation of plasma exchange hypoglycemia resolved, and glucose lability improved.