Papillary renal cell‐like carcinoma in a retroperitoneal teratoma

We report a case of somatic type malignancy with papillary renal cell carcinoma differentiation arising in a retroperitoneal mixed germ cell tumor. The patient was a 36‐year‐old man with a synchronous mediastinal teratoma. The somatic type malignancy in the retroperitoneal tumor was composed of papillary structures covered by atypical epithelial cells with eosinophilic cytoplasm, prominent nucleoli and pseudostratified nuclei. Papillary cores contained numerous aggregates of foamy macrophages, typical of type I papillary renal cell carcinomas. The immunohistochemical profile was consistent with papillary renal cell carcinoma, including positive reactions for cytokeratin 7 and alpha‐methyl acyl CoA racemase. There was no somatic type malignancy component in the synchronous mediastinal teratoma. Both the retroperitoneal and the mediastinal tumor showed gains of 12p and chromosome 17 material. There was no c‐MET mutation in the somatic type malignancy. To our knowledge, this is the first report of a somatic type malignancy with features of papillary renal cell carcinoma arising in a germ cell tumor. It is important not to confuse such a retroperitoneal tumor with a conventional papillary renal cell carcinoma, because presence of other malignant histologies within the germ cell tumor may warrant different treatment. In such cases, the presence of isochromosome 12p can be helpful to the diagnosis.     

Effect of sleep deprivation on multi‐unit discharge activity of basal forebrain

The basal forebrain (BF) is an important wakefulness/arousal‐promoting structure involved in homeostatic responses to sleep deprivation (SD). However, the effects of SD and subsequent sleep recovery on the BF discharge have not been investigated. Multi‐unit BF activity was recorded on freely moving rats during 8 h of baseline (BL) and, on the following day, during 4 h of SD by gentle handling followed by 4 h of recovery. The effect of SD on the waking discharge was evaluated during the last 10 min of each hour when attentive waking was induced. The wakefulness level was defined based on the ratio between theta and delta electroencephalogram (EEG) powers, and epochs with ratios ≥1 but <2 (T/D‐1) and ≥2 but <4 (T/D‐2) were analysed separately. During T/D‐1 wakefulness, the BF multi‐unit discharge rate increased significantly during the second and third hours of SD and decreased during the third hour of recovery when compared with corresponding hours of BL. Non‐rapid eye movement sleep discharge rate during recovery decreased significantly in the second and third versus the first and last hours. The results suggest that maintenance of the level of vigilance necessary for adequate performance during SD requires increased activation of BF neurones when compared with the BL, whereas the same level of vigilance after several hours of recovery can be maintained with lesser activation of BF neurones.     

PCR引物浓度对寡核苷酸液相杂交的影响

目的考察PCR引物浓度配比对液相微珠杂交效率的影响,寻求具有较强杂交信号和较好稳定性的PCR引物浓度配比。方法建立HLA-DRB1等位基因的相关数据库,选择在HLA-DRB1位点的第二外显子上设计探针,并且选择其保守序列作为阳性对照探针(DPC2),DPC2探针中间位点T突变成A作为阴性对照探针(DNC)。分别针对标本C2-008、C2-024、C2-025的等位基因序列设计出6条约21bp的寡核苷酸探针,各探针5’端用氨基(NH2)修饰。通过引物浓度梯度配比(1:100、1:50、1:20、1:8、1:4、1:2、1:1),对型别已知的细胞株DNA进行PCR扩增并得到目的片段(1:100配比除外),在相同条件下将PCR产物与寡核苷酸探针进行液相杂交检测。结果浓度配比为1:1的对称式扩增产物杂交结果不理想,而浓度配比分别为1:20、1:8、1:4、1:2的不对称扩增均得到了待检单链、双链DNA混合物,其中1:4浓度配比具有最好的扩增效率和稳定性。根据阳性信号与阳性标本是否相符表明:引物浓度配比为1:100的不对称PCR和1:1的对称PCR检测效果差,易出现假阴性;1:2、1:4、1:8、1:20配比检测效果较好,比较稳定。结论PCR引物浓度配比影响液相微珠杂交效率,不对称PCR产物有利于提高杂交效率,为快速成功配制PCR试剂奠定了基础,有利于寡核苷酸液相芯片的应用。     

Neonatal alloimmune thrombocytopenia caused by human leucocyte antigen-B27 antibody

Neonatal alloimmune thrombocytopenia (NAIT) occurs when maternal alloantibodies to antigens presented on foetal platelets cause their immune destruction. Whether human leucocyte antigen (HLA) antibodies can cause NAIT is controversial. Here, a patient was described who suffered from a NAIT caused by an HLA-B27 antibody. Sera from the mother and the newborn were tested for human platelet antigen antibodies and HLA antibodies by monoclonal antibody-specific immobilization of platelet antigens (MAIPA) assay, solid phase-linked immunosorbent assay (ELISA), lymphocytotoxicity assay (LCT) and flow cytometric analysis. No antibodies against cluster designation (CD)109 and platelet glycoproteins of the father were found in patient's and mother's serum. However, HLA ELISA was used to identify HLA antibody in both sera. The antibody was specified as HLA-B27 antibody. Typing results showed that the father descended HLA-B27 antigen on patient and his brother. The mother was HLA-B27 negative. It is most conceivable that the previous pregnancy of the mother induced the production of anti-HLA-B27 antibody, which crossed the placenta and subsequently caused an NAIT in the case presented.     

Characterisation of dic(9;20)(p11-13;q11) in childhood B-cell precursor acute lymphoblastic leukaemia by tiling resolution array-based comparative genomic hybridisation reveals clustered breakpoints at 9p13.2 and 20q11.2

Although the dic(9;20)(p11-13;q11) is a recurrent chromosomal abnormality in paediatric B-cell precursor acute lymphoblastic leukaemia (BCP ALL), occurring in approximately 2% of the cases, its molecular genetic consequences have not been elucidated. In the present study, high-resolution genome-wide array-based comparative genomic hybridisation (array-CGH) and fluorescence in situ hybridisation (FISH) were used to characterise the 9p and 20q breakpoints (BPs) in seven childhood BCP ALLs with dic(9;20), which was shown to be unbalanced in all of them, resulting in loss of 9p13.2-pter. Five of the cases had loss of 20q11.2-qter, whereas two displayed gain of 20cen-pter. All BPs on 9p clustered in a 1.5 Mb segment of the sub-band 9p13.2; in three of the cases, the 20q BPs mapped to three adjacent clones covering a distance of 350 kb at 20q11.2. Thus, the aberration should be designated dic(9;20)(p13.2;q11.2). One of the ALLs, shown to have a complex dic(9;20), was further investigated by FISH, revealing a rearrangement of the haemapoietic cell kinase isoform p61 (HCK) gene at 20q11. The disruption of HCK may result in a fusion gene or in loss of function. Unfortunately, lack of material precluded further analyses of HCK. Thus, it remains to be elucidated whether dic(9;20)(p13.2;q11.2) leads to a chimaeric gene or whether the functionally important outcome is loss of 9p and 20q material.     

Detection of H. pylori antibody profile in serum by protein array

AIM: To detect multiple H pylori antibodies in serum samples of individuals who carryH pylori by protein array. METHODS: Recombinant H pylori antigens, urease B subunit (UreB), vacuolating toxin A (VacA) and cytotoxin associated gene A protein (CagA), were prepared and immobilized in matrixes on nitrocellulose membrane by robotics to bind the specific immunoglobulin G (IgG) antibodies in serum. Staphylococcus protein A (SPA) labeled by colloid gold was used to integrate the immuno-complex and gave red color signal. The scanner based on charge-coupled device (CCD) could collect the image signal and convert it into digital signal. RESULTS: When human IgG was printed on the membrane in increasing concentrations and incubated with immunogold, a linear dose response curve was obtained and the detection limit for IgG was about 0.025 ng. The cutoff values, which were defined as the mean grey level plus 3 times of standard deviation, were 27.183, 28.546 and 27.402, for anti-UreB IgG, anti-CagA IgG and anti-VacA IgG, respectively, as 400 human serum samples with negative H pylori antibodies were detected by the protein array. When 180 serum samples from patients in hospital were employed for detection of IgG against UreB, CagA and VacA, the sensitivity of the protein array was 93.4%, 95.4%, 96.0%, and the specificity was 94.8%, 94.4% and 97.5%, respectively, as compared with the results obtained by ELISA. The assay also showed high reproducibility, uniformity and stability, and the results were available within 30 min. CONCLUSION: The protein array is a very practical method for rapid detection of multiple antibodies in serum samples. It is especially useful for large scale epidemiological investigation of the infection of H pylori.     

拉米夫定耐药突变相关慢性重型乙型肝炎27例分析

目的 研究在拉米夫定治疗慢性乙型肝炎的过程中出现拉米夫定耐药突变相关的慢性重型乙型肝炎的临床特点.方法 回顾性分析27例在拉米夫定治疗过程中出现耐药突变的慢性重型乙型肝炎病例,分析其临床特征,用基因芯片法或基因测序法检测YMDD突变类型,对8例施行肝移植术后的离体肝组织分析其病理特点.采用χ2检验进行统计学分析.结果 27例拉米夫定耐药患者的YMDD突变类型分别为:YVDD突变5例,YVDD突变+L180M突变2例,YIDD突变13例,YIDD+L180M突变4例,YVDD突变+YIDD突变1例,YVDD突变+Y1DD突变+L180M突变2例,单独L180M突变未检出.根据治疗前是否存在肝硬化,分为肝硬化组和非肝硬化组.与肝硬化组相比,非肝硬化组慢性重型乙型肝炎的发生率低,预后好,年龄小,HBeAg阳性率高.8例离体肝组织病理分析显示两种主要病理类型,一种主要表现为活动性肝硬化,另一种为大块或亚大块坏死,肝脏显著萎缩.结论 肝硬化为拉米夫定耐药突变相关的慢性重型乙型肝炎的高危因素.拉米夫定耐药突变相关的慢性重型乙型肝炎可能存在两种发病机制.     

基因芯片技术分析川芎嗪对小鼠实验性结肠炎的影响

目的 用基因芯片技术研究川芎嗪对实验性结肠炎小鼠基因表达谱的影响.方法 健康昆明小鼠30只,均分为对照组、0.9%氯化钠组及川芎嗪组.除对照组外,其余小鼠均以恶唑酮灌肠造模.提取0.9%氯化钠组及川芎嗪组结肠组织mRNA,制备cDNA探针,分别用Cy3和Cy5两种荧光染料标记,与基因表达谱芯片进行杂交,经扫描、分析,得出药物作用后表达有差异的基因,并应用荧光定量RT-PCR技术对其中2个表达差异明显的基因(白细胞介素-10和白细胞介素-4)进行验证.结果 川芎嗪组差异表达基因432个,占芯片基因总数的2.86%.其中表达上调的基因307个,表达下调的基因125个,其中部分基因已知其功能.结论 川芎嗪通过抑制肠道炎性反应、抗粘附分子及改善免疫功能等多种作用途径为治疗溃疡性结肠炎提供了理论上的可能,但更为精细的治疗靶点还有待进一步探索.     

强直性脊柱炎患者外周血T细胞亚群的变化及意义

目的探讨强直性脊柱炎（AS）患者外周血中T细胞亚群的变化及其意义。方法采用流式细胞仪（FCM）检测外周血淋巴细胞表面CD3、CD4、CD8及其胞质内细胞因子IFN-γ和IL-4的表达。结果与正常对照组相比,As患者外周血Th细胞（CD;CD4＋）百分率无显著差异,Tc细胞（CD;CD;）明显升高（P〈0．05）;Th1细胞（CD3^＋CD4^＋IFN-γ^＋）百分率明显升高,Th2细胞（CD3^＋CD4^＋IL4^＋）无显著性差异;Te1细胞（CD3^＋CD8^＋IFN-γ^＋）百分率明显降低,T02细胞（CD3^＋CD8^＋IL4^＋）无显著性差异。结论AS患者外周血T细胞Th1／Th2、Tcl／Tc2亚群比例失衡,呈Th1、Tc2优势型。     

Transient reduction of spontaneous neuronal network activity by sublethal amyloid β (1–42) peptide concentrations

Soluble amyloid β_1–42 (Aβ_1–42) peptide has recently been assigned a key role in early Alzheimer’s disease (AD) pathophysiology accounting for synaptic dysfunction before amyloid plaque formation and neurodegeneration can occur. Following sublethal Aβ_1–42 administration, we observed an acute but transient reduction of the spike and burst rate of spontaneously active cortical networks cultured on microelectrode arrays. This simple experimental system appears suitable for future long-term pharmacological and genetic studies of Aβ_1–42 signaling, thus providing a valuable new tool in AD research.