Identification of mutations in the Bruton's tyrosine kinase gene,including a novel genomic rearrangements resulting in large deletion,in Korean X-linked agammaglobulinemia patients

Mutations in the Bruton's tyrosine kinase (BTK) gene are responsible for X-linked agammaglobulinemia (XLA). We identified BTK mutations in six patients with presumed XLA from unrelated Korean families. Four out of six mutations were novel: two missense mutations (P565T, C154Y), a point mutation in a splicing donor site (IVS11+1G>A), and a large deletion (a 6.1-kb deletion including BTK exons 11–18). The large deletion, identified by long-distance PCR, revealed Alu-Alu mediated recombination extended from an Alu sequence in intron 10 to another Alu sequence in intron 18, spanning a distance of 6.1 kb. The two known mutations consisted of one missense (G462D) mutation, and a point mutation in a splicing acceptor site (IVS7−9A>G). This study suggests that large genomic rearrangements involving Alu repeats are few but an important component of the spectrum of BTK mutations.     

Epithelioid variant of gastrointestinal stromal tumor: Diagnosis by fine-needle aspiration

Epithelioid gastrointestinal stromal tumors (GISTs) may cause significant diagnostic confusion on fine-needle aspiration (FNA) with carcinomas, neuroendocrine tumors, and melanoma, particularly when metastatic. This study characterizes the cytologic features of nine cases of epithelioid GISTs that were obtained by computerized tomographic guidance in five, by endoscopic ultrasound in three, and from an excised liver tumor in one. Six cases presented as liver masses, one as a perisplenic mass, one as an abdominal mass, and one as a gastric mass. The aspirates revealed mainly single or small clusters of epithelioid cells with a moderate amount of granular to clear cytoplasm, small uniform nuclei with mild to marked nuclear envelope irregularities. Binucleation and intranuclear inclusions were frequent findings. Collagenous stroma was seen in most cases. In three cases, a neuroendocrine tumor was the initial diagnosis. Immunocytochemical staining for c-kit (CD117) was performed on cellblocks in six cases and was positive in five cases. On the subsequent surgical specimen, CD117 was positive in the c-kit-negative cytology case. The diagnosis of GIST should be considered in aspirates of the gastrointestinal tract, liver, mesentery, or abdominal wall mass lesions when epithelioid cells are the predominant cell type. Ancillary studies such as immunohistochemical stains are usually helpful in making a definitive diagnosis.     

从事医疗放射工作人员遗传损伤的研究

目的 了解温州地区医疗放射人员的细胞遗传学变化。方法 以外周血淋巴细胞培养法作微核检测。结果 放射组微核率为3.425‰,对照组微核率为1.06‰,两者有显著性差异(P<0.05)。结论 医疗放射工作者存在一定程度 的遗传损伤。     

Kinetics of severe acute respiratory syndrome (SARS) coronavirus-specific antibodies in 271 laboratory-confirmed cases of SARS

The sensitivities and specificities of an immunofluorescence assay and an enzyme immunoassay for detection of antibodies specific for severe acute respiratory syndrome coronavirus (SARS-CoV) were compared for 148 laboratory-confirmed SARS cases. The appearance and persistence of SARS-CoV-specific antibodies were assessed, with immunoglobulin G detected in 59% of samples collected within 14 days and persisting for 60 to 95 days after the onset of illness.     

Origins of tooth pulp-evoked far-field and early near-field potentials in the cat

Electrical stimulation of the mandibular canine tooth pulp in barbiturate-anesthetized cats activated myelinated A-beta-fibers in the inferior alveolar (dental) nerve and evoked a complex pattern of short-latency potentials that were recorded epidurally from the cortex. Three far-field components ( FFPs designated I, IIa,b, and IIIa,b) and two early near-field components ( ENFPs designated P1 and N1) were identified from computer-averaged potentials recorded over the contralateral lateral sigmoid gyrus or ipsilateral presylvian /anterior coronal gyri. Several corroborative lines of evidence indicated that tooth pulp-evoked FFPs and ENFPs originate from the following generator sources: I, inferior alveolar (dental) nerve/semilunar (gasserian) ganglion; IIa,b, trigeminal lemniscal fibers from the principal (main) sensory nucleus; IIIa,b, thalamocortical fibers from the thalamic ventral posteromedial nucleus; P1, N1, cytoarchitectural area 3 on the banks of the coronal or orbital sulcus. The latencies and waveforms of volume-conducted, surface-recorded potentials were not significantly different from those recorded at putative generator sites along the trigeminal lemniscal projection system. The short latencies and distribution of surface- and depth-recorded tooth pulp-evoked potentials suggested bilateral, trisynaptic paths from peripheral nerve to cortical neurons. Amplitude measurements of FFPs and ENFPs recorded at the cortical, epidural surface were compared to calculated values based on a mathematical model of two concentric spheres of inhomogeneous media in which a single equivalent dipole is radially oriented at some distance from the center. The morphology and distribution of FFPs and ENFPs are discussed in relationship to source geometry. Surface- and depth-recorded tooth pulp-evoked potentials had recovery functions that were covariant with rates of stimulation. In addition, stimulation of putative generator sites along the trigeminal lemniscal system reproduced portions of the tooth pulp-evoked surface-recorded potentials. Selective ablation of trigeminal cortical area 3 abolished ENFPs and lesions of lemniscal brain stem sites abolished FFPs . Disruption of extralemniscal brain stem sites had no effect on the surface-recorded evoked potentials examined. These findings and other supporting evidence from previous studies imply that the projection of fast-conducting tooth pulp input to the cortex may subserve a nonnociceptive modality and participate in sensorimotor integration.     

IL-10RA基因多态性及其与系统性红斑狼疮关系的研究

目的:确定SLE模型小鼠IL-10RA基因变异及其与SLE表现型是否存在关联。方法:用微卫星遗传标记及数量性状位点(QTL)分析方法确定SLE模型小鼠B/W F1的SLE易感基因精确染色体定位并选取候选易感基因,对候选易感基因进行测序分析,选取有基因序列异常的候选易感基因进行PCR-SSCP分析,确定候选易感基因碱基序列变异位点与抗染色质抗体、抗DNA抗体,抗组蛋白抗体及蛋白尿等SLE表现型的相关关系。结果:QTL分析结果表明B/W F1×NZB小鼠抗染色质抗体易感基因与NZW型IL-10RA基因紧密连锁;测序分析发现IL-10RA基因编码区有18处碱基变异,其中7处碱基变异将导致编码氨基酸的变异;抗染色质抗体、抗DNA抗体,抗组蛋白抗体及蛋白尿等SLE表现型与NZW型IL-10RA基因密切相关。另一种SLE模型小鼠MRL的IL-10RA基因存在相同变异。结论:NZW小鼠IL-10RA基因编码区碱基序列存在变异,B/W F1×NZB小鼠SLE表现型与NZW小鼠第9染色体IL-10RA编码区碱基变异相关,提示IL-10RA可能是SLE模型小鼠的一个SLE易感基因。     

单肺移植手术前后的供体肺保护

目的 探讨肺移植手术前后供体肺保护的处理经验.方法 回顾性分析了2003年1月至2006年8月本院施行的同种异体单肺移植手术9例,其中左侧3例,右侧6例.冷低钾肺保护液灌注移植肺,术后三联免疫抑制剂抗排斥,并分析术后的胸片评分、氧合指数及肺动脉压变化情况.结果 供体肺缺血时间平均为(308.6±50.8)min.8例患者均成功脱离呼吸机并存活过围手术期(＞30 d),术后拔除气管插管时间平均(4.8±3.4)d,1例未能脱机并在术后22 d死亡.结论 肺保护需要在多个环节中加以控制,不同时期的保护侧重点不同.     

交链孢酚激活人胎食管上皮组织细胞癌基因的研究

本文报道食管癌高发区粮食中优势污染菌互隔交链孢霉代谢产物交链孢酚(AOH)对人胎儿食管上皮组织癌基因的激活。结果表明,体外培养的人胎儿食管上皮组织经10gg/ml AOH作用4h后,其DNA具有转化活性,可转化NIH/3T3细胞。一轮转化率为0.17Foci/μg DNA,二轮转化率为0.58Foci/μg DNA(P<0.01)。转化细胞中含有人特异性高重复序列Alu,提示转化系人DNA转染所致。Southern blot检测表明,AOH处理后的人胎儿食管上皮组织内有H-ras和myc基因的扩增,经其转染的NIH/3T3细胞中亦有H-ras基因持久而稳定的扩增。对照组胎儿食管上皮和正常NIH/3T3细胞中均未发现相应的同源序列,说明正常胎儿食管组织中的H-ras和myc基因可经AOH短期处理而激活。该结果对AOH可能系人食管癌病因之一提供了直接证据。     

RAI16蛋白合成肽多克隆抗体的制备及初步应用

目的: 制备RAI16蛋白的合成肽多克隆抗体, 并进行初步鉴定和应用, 为研究RAI16蛋白的功能及作用机制获得重要的实验工具.方法: 应用Fmoc法化学合成RAI16蛋白N端第44～55位氨基酸的多肽, 经C18的RP-HPLC纯化后, 通过高碘酸钠法将纯化的RAI16蛋白的多肽与KLH交联; 皮下注射抗原免疫新西兰纯种大耳白兔, 加强免疫得到抗血清, 应用蛋白G纯化获得多克隆抗体.对纯化的抗体进行ELISA、免疫组化、 Western blot等初步鉴定和应用.结果: 化学合成RAI16蛋白N端第44～55位氨基酸的多肽, 纯化后多肽纯度为96% , 达到免疫用抗原标准.多肽与KLH交联, 用于免疫动物.经纯化后的抗体效价为1∶ 125 000.该多肽抗体可特异识别人脾脏组织中相对分子质量(Mr)约为55 000的RAI16蛋白.结论: 所制备的多克隆抗体能与天然RAI16蛋白发生特异性反应, 可应用于ELISA、免疫组化、免疫沉淀和Western blot等实验, 为确定RAI16蛋白的组织分布和亚细胞定位、研究RAI16蛋白的功能及作用机制提供了重要的实验工具.     

川崎病休克综合征合并心力衰竭1例临床病例分析及文献复习

目的 探讨川崎病休克综合征 (Kawasaki Disease Shock Syndrome,KDSS) 合并心力衰竭的临床特征。方法 分析 1例KDSS合并心力衰竭患儿的临床资料,并复习相关文献。结果 KDSS在川崎病中不常见,但表现严重,在急性期可以导致循环衰竭,表现为心动过速、毛细血管充盈时间延长、四肢末端发凉、脉搏细弱、尿量减少或意识障碍等而发生全身循环衰竭,早期识别,及时给予人免疫球蛋白和激素等综合治疗可明显改善患者休克的状况。结论 KDSS是川崎病中少见但较为严重的临床并发症,早期识别及综合的临床治疗方法是减少KDSS的有效措施。