MACS检测胃癌腹腔冲洗液游离癌细胞的研究

目的 探讨胃癌腹腔微转移情况的检测方法及意义。方法 手术中切除肿瘤前收集腹腔冲洗液，采用磁激活细胞分离术（MACS）对不同病理分期胃癌患者腹腔冲洗液中癌细胞进行富集并检测。分别标记带磁珠的细胞角蛋白（CK）抗体，经磁柱富集CK^＋上皮细胞，用流式细胞仪检测其含量，并比较胃癌组与胃平滑肌瘤组（对照组）以及胃癌不同分期之间、磁富集前后CK^＋上皮细胞含量的差异。结果 在未经MACS富集的标本中较少发现CK^＋ CD45^-细胞；在富集后的标本中其含量在胃癌组与对照组有显著差异（41／50，1／10，P〈0．001）；pTNMⅠ～Ⅱ期与Ⅲ～Ⅳ期之间（0．67％，3．42％，P〈0．001）差异有非常显著性。结论 MACS能有效地富集上皮来源细胞，提高上皮源细胞的检出率，并能反映腹腔游离癌细胞数量；上皮细胞数量与胃癌的存在及临床病理分期有关，其有利于判断肿瘤转移和预后并指导治疗。     

Protein sorting within the mhc class II antigen-processing pathway

Major histocompatibility complex (MHC) class II molecules are required for the presentation of antigenic peptides that are derived predominantly from internalized proteins. The assembly of MHC class II/peptide complexes occurs within endosomal compartments of antigen-presenting cells (APCs). Therefore, for assembly to occur, MHC class II molecules, foreign proteins, and accessory molecules must be sorted to appropriate intracellular sites. My laboratory is trying to understand how proteins are sorted to various antigen-processing compartments as well as to conventional endosomal organelles. Using chimeric marker proteins and a variety of biochemical and genetic approaches, we are addressing the specificity of protein sorting and the mechanisms by which sorting signals are deciphered. By using a similar chimeric protein approach to target endogenous proteins to distinct compartments, we hope to address the role of processing events in each compartment in the generation of MHC class II ligands.     

Melanosomes and MHC class II antigen-processing compartments

Melanosomes are specialized intracellular compartments within melanocytes and retinal pigment epithelial cells that function in the synthesis, storage, and secretion of melanins, which are the major pigments made by mammals. The mechanisms that regulate the formation of melanosomes, and the pathways by which constituent proteins are targeted to them, are related to those involved in the biogenesis of major histocompatibility complex (MHC) class II antigen-processing compartments. Consequently, diseases that affect pigmentation may also affect antigen presentation to T cells. Moreover, many of the tissue-specific proteins that localize to melanosomes and participate in melanin formation double as tumor-associated antigens that are targets for T cells in patients with melanoma. Our studies on melanosome biogenesis are providing new ways of thinking about antigen-processing compartments and the mechanisms regulating presentation of tumor-associated antigens.     

A study of visual and auditory verbal working memory in schizophrenic patients compared to healthy subjects

Impaired working memory (WM) performance is considered as a central feature of schizophrenia. Divided into two components, verbal and spatial, WM has been shown to involve frontal and parietal regions. Verbal WM can be tested either visually or aurally. The present study aimed to test schizophrenic patients in both visual and auditory verbal WM in order to assess a possible distinct pattern of alteration of these two modalities. Twenty-four schizophrenic patients and 24 healthy controls were compared with 2-back continuous visual and auditory verbal WM testing. Both groups were also tested on a neuropsychological battery including Wisconsin Card Sorting Test (WCST). Schizophrenic patients were less efficient in both verbal WM tests. When taking age and educational level as covariates and both WM modalities as dependent variables, there was no differential effect of modalities across groups. In further exploratory analyses, partial correlations brought association between verbal WM and psychosocial adaptation, WCST and length of illness. These results suggest a similar pattern of alteration of both modalities of verbal WM in schizophrenic patients. The implications of this finding are discussed. Received: 16 February 1999 / Accepted: 20 December 1999     

多元联系数在医院医疗质量综合评价排序中的应用

目的通过把多元联系数用于医院医疗质量综合评价排序的探讨,为医院医疗质量综合评价提供一种新的简便实用方法.方法根据评价对象个数m选用m元联系数,对m个评价对象的每个指标数据从优到劣编秩次从1到m,计算每个指标数据与该指标最优值的同一度,再根据秩次和m元联系数联系分量相对应的规则写出每个评价对象的m元联系数表达式,通过联系分量系数的取值把m元联系数转化为联系数值,根据联系数值的大小作出综合评价排序结论.结果利用m元联系数可以使综合评价计算过程简化,而评价和排序结果不变.结论可以在医院医疗质量综合评价排序中应用多元联系数.     

Variants in the UBR1 gene are not associated with chronic pancreatitis in Japan

Background/objectivesThe UBR1 gene encodes the enzyme ubiquitin-protein ligase E3 component n-recognin 1. Loss-of-function mutations in the UBR1 gene cause Johanson-Blizzard syndrome, which involves pancreatic exocrine insufficiency. No previous studies have examined an association of UBR1 variants with pancreatitis, in part due to the large size of the gene. This study aimed to clarify whether UBR1 variants are associated with chronic pancreatitis (CP) by the application of targeted next generation sequencing.MethodsExon sequences of the UBR1 gene from 389 Japanese patients with CP (188 idiopathic, 172 alcoholic, 20 hereditary, 9 familial) were captured by the HaloPlex target enrichment technology and subjected to next generation sequencing.ResultsNinety nine point two % of the coding regions of the UBR1 gene could be sequenced by ≥ 20 reads with a mean read depth of 595 and a median depth of 399. Fifteen non-synonymous variants including three novel ones [c.4514T > C (p.I1505T), c.4828C > G (p.H1610D) and c.4856A > T (p.D1619V)] and two synonymous variants were identified in the exonic regions. The frequency of any non-synonymous or synonymous variants was not different between the patients with CP and controls.ConclusionsVariants in the UBR1 gene were not associated with CP in Japan.     

Cognitive inflexibility in Japanese adolescents and adults with autism spectrum disorders

AIM: To investigate executive function in Japanese adolescents and adults with autism spectrum disorders(ASD) compared to Japanese controls.METHODS: Thirty-three individuals with ASD and 33 controls participated. The ASD and control groups' demographic variables were matched for gender(male/female: 20/13 vs 20/13), age(26.1 ± 11.5 vs 26.8 ± 9.6), years of education(13.2 ± 2.9 vs 14.2 ± 1.9), full-scale intelligence quotient(IQ)(103.0 ± 16.7 vs 103.7 ± 14.7), performance IQ(96.2 ± 16.1 vs 97.8 ± 15.0), and verbal IQ(107.9 ± 16.3 vs 107.7 ± 14.4). Participants performed the Wisconsin Card Sorting Test(WCST), which assesses the executive processes involved in problem solving and cognitive flexibility, and the Continuous Performance Test(CPT), which assesses attention and impulsivity. Symptoms were assessed by the Autism-Spectrum Quotient Japanese version(AQ-J). First, we compared the scores of the WCST between the groups using a Mann-Whitney U-test and conducted an analysis of covariance for the variables with the scores of category archives and CPT scores as covariates. Second, we analyzed the correlation between the scores of the WCST and the AQ-J in the ASD group using Pearson's r.RESULTS: The total errors(TE) and the percentages of perseverative errors of the Milner type(%PEM) and Nelson type(%PEN) among the TE in the ASD group were significantly worse compared with the control group(ASD vs Control, respectively: TE: 16.0 ± 6.2 vs 12.6 ± 3.5, P = 0.012; %PEM: 11.7 ± 10.7 vs 6.6 ± 8.9, P = 0.037; %PEN: 20.1 ± 14.5 vs 8.7 ± 10.4, P = 0.0011). In contrast, no significant difference was observed between the two groups in the scores of categories achieved on the WCST or the CPT. An analysis of covariance revealed significant differences between the groups in the %PEN scores(P = 0.0062) but not in the TE or the %PEM scores. These results suggest that Japanese adolescents and adults with ASD have cognitive inflexibility. Furthermore, our results suggest that Japanese adolescents and adults with ASD may have difficulties using negative feedback because perseverative errors of the Nelson type indicate persistence in choosing the incorrect reaction. By contrast, there was no significant correlation between the WCST and AQ-J scores.CONCLUSION: We confirmed the presence of cognitive inflexibility in Japanese adolescents and adults with ASD. Our results also indicated that subjects with ASD may not use negative feedback effectively.     

威斯康星卡片分类测验作为双生子基因功能稳定性指标的初探

目的：探索威斯康星卡片分类测验测试各指标中更稳定、更精确的指标（内表型）。方法：在重庆市主城区募集6—16岁的双生子。签写知情同意书后，用威斯康星卡片分类测验对59对6—16岁的双生子测试（同卵双生28对，异卵双生31对），比较双生子两个体之间的六个常用指标（正确数、错误数、持续错误数、非持续错误数、总分类数、完成第一个分类所需个数）得分的相关系数，采取双生子的颊黏膜标本以提取DNA并进行卵型鉴定。结果：MZ组和DZ组的年龄、性别、受教育年限具有可比性（P〉0．05），MZ组的持续错误数指标在双生子对个体之间的相关系数有显著相关（r=0．65，P=0．001），其余五个指标和DZ组的六个指标在双生子对个体之间的得分无显著性相关（P〉0．05）。结论：在威斯康星卡片分类测验常用的六个指标中，持续错误数受遗传的影响更大，作为内表型指标可能优于威斯康星卡片分类测验中的其他指标。     

经流式细胞仪分选神经细胞技术方法的建立

目的:探索建立一种分离在体模型中不同种类神经细胞的技术方法。方法:采用免疫荧光标记方法和流式细胞仪分选技术。结果:NeuN和GFAP双标显色可见,GFAP阳性的星形胶质细胞无NeuN表达,经过流式细胞仪分选可获得高纯度的星形胶质细胞和神经元,其阳性标记率分别为31.91%±4.67%和47.70%±6.30%。结论:该分选方法可以同时获得纯度高、数量多的不同种类的神经细胞。