多源耐药基因表达水平的检测在儿童白血病中的临床意义

应用逆转录ＰＣＲ结合同位素定量分析，对３２例儿童白血病患者的多源耐药基因表达水平进行了研究。结果显示，初发病人的表达均较低，复发病人表达较高，缓解期病人表达程度介于两者之间，为探讨多源耐药基因表达水平与临床化疗之间的相关性及逆转克服多源耐药性药物的应用，提供了一定的理论依据。     

Point mutation in the parkin gene on patients with Parkinson’s disease

Summary To investigate the distribution of possible novel mutations from parkin gene in variant subset of patients with Parkinson’s disease (PD) in China and explore whether parkin gene plays an important role in the pathogenesis of PD, 70 patients were divided into early-onset group and late-onset group; 70 healthy subjects were included as controls. Genomic DNA from 70 normal controls and from those of PD patients were extracted from peripheral blood leukocytes by using standard procedures. Mutations of parkin gene (exon 1–12) in all the subjects were screened by PCR-single strand conformation polymorphism (SSCP), and further sequencing was performed in the samples with abnormal SSCP results, in order to confirm the mutation and its location. A new missense mutation Gly284Arg in a patient and 3 abnormal bands in SSCP electrophoresis from samples of another 3 patients were found. All the DNA variants were sourced from the samples of the patients with early-onset PD. It was concluded that Parkin point mutation also partially contributes to the development of early-onset Parkinson’s disease in Chinese. WANG Tao, male, born in 1961, Associate Professor This work was supported by grants from the key program of the special scientific project of Scientific & Technologic Agency of Hubei Province (Serial No. 2001AA308B01) and the Hygienic Research Project of Hygienic Agency of Hubei province (Serial No. WJ 01529).     

Sry监测体外扩增造血细胞植入效果的实验研究

目的：探讨Y染色体性别决定基因(Sry)作为评价和追踪造血细胞移植后植入状态检测指标的可行性。方法：根据Sry DNA序列设计引物及制备探针，然后进行PCR检测、斑点杂交和原位杂交．动态追踪经不同条件下体外扩增的纯系雄性小鼠BMMNC回输至雌性小鼠后的植入状态。结果：PCR结果显示在各移植组小鼠的骨髓有核细胞、脾细胞及外周血白细胞中均有Y染色体特异性序列的存在；斑点杂交结果显示在各回输组间并无明显差别，但用脾脏组织作原位杂交的结果则发现基质细胞支持扩增回输组的阳性颗粒数目与输注新鲜细胞组相似，但略少于雄性小鼠；而输注细胞因子扩增细胞实验组小鼠则明显少于雄性动物的阳性对照标本和基质细胞支持扩增回输组。结论：(1)经重复检测表明上述方法的重复性好，结果可信．可作为检测性别不同时造血干细胞移植效果的一种检测手段；(2)证实基质细胞支持下的体外扩增的造血细胞具有较好的植入能力。     

Testicular amplificaion and impaired transmission of human butyrylcholinesterase cDNA in transgenic mice

Gene amplification occurs frequently in tumour tissues yet is,in general, non-inheritable. To study the molecular mechanismsconferring this restraint, we created transgenic mice carryinga human butyrylcholinesterase (BCHE) coding sequence, previouslyfound to be amplified in a father and son. Blot hybridizationof tail DNA samples revealed somatic transgene amplificationswith variable restriction patterns and intensities, suggestingthe occurrence of independent amplification events, in 31% (11/35)of mice from the FII generation but in only 3.5% (2/58) of theFII and FIV generations. In contrast, >10-fold amplificationsof the BCHE transgene and the endogenous acetylcholinesteraseand c-raf genes appeared in both testis and epididymis DNA from>80% of FIII mice. Drastic, selective reductions in testisBCHEmRNA but not in actin mRNA were detected by the PCR amplificationof testis cDNA from the transgenic mice, and apparently resultedin the limited transmission of amplified genes. The testicularamplification of the BCHE transgene may potentially representa general phenomenon with clinical implications in human infertility.     

人抗HBsAg噬菌体抗体Fab段基因的序列分析及表达

对已建的噬菌体抗体库分离出来的人抗－ＨＢｓ克隆进行了序列分析和表达研究，发现４个克隆中３个克隆的重链和轻链完全相同，ＤＮＡ序列分析表明ＶＨ分别属于ＶＨ１亚群和Ⅱ亚群，其轻链ＶＬ分别属于ＶλⅡ亚群和ＶλⅠ亚群。构建了可溶性Ｆａｂ段表达载体，显示出在细菌中表达的Ｆａｂ段抗体与ＨＢｓＡｇ特异性结合，这说明所筛选出来的噬菌体抗体具有ＨＢdisplay status     

慢性乙型肝炎表面抗原携带者前S1基因的高度异质性

应用半巢式聚合酶链式反应（ＰＣＲ）从一例慢性ＨＢｓＡｇ携带者血清中扩增出ＨＢＶ前Ｓ１基因，将其克隆于噬菌体Ｍ１３ｍｐ１９中进行序列分析。结果发现：与同源性最好的ＨＢＶａｄｒ野生林相比，所测的１０个克隆均有替代和插入突变，９个克隆有缺失突变，１０个克隆的核苷酸变异率为５．０％－１７．０％，氨基酸的变异率为１３．０－６０．０％；１０个克隆之间的核苷酸变异率为２．３％－２４．３％，氨基酸的变异率为１４．     

Terminal deletion of chromosome 4p (4p16.3) shows a breakpoint between loci linked to Huntington disease

A 15-year-old boy with a terminal deletion of the short arm of chromosome 4 is described. The patient has a mild clinical phenotype that is incompatible with Wolf-Hirschhorn syndrome. Careful neurological examination including CT scan did not show any signs of Huntington disease. The chromosomal breakpoint was analyzed by means of polymorphic DNA probes localized close to the tentative Huntington (HD) locus. The breakage has occurred between D4S43 and D4S90 loci and thus deletes part of the chromosomal candidate regions for the HD locus. © 1992 Wiley-Liss, Inc.     

E1 Tor霍乱弧菌SM_6和江苏1d菌株

ＳＭ６是我国检测Ｅ１Ｔｏｒ霍乱弧菌是否带有溶源性噬菌体的指示菌株。江苏的６株Ｅ１Ｔｏｒ霍乱弧菌和ＳＭ６都属于噬菌体－生物分型的１ｄ型，而在噬菌体分型中ＳＭ６为１／２型，江苏的６株菌株则为１／２／３型；霍乱毒素（ＣＴ）基因测定也表明两者的不同，ＳＭ６不含ＣＴ基因，江苏的６株都含有ＣＴ基因，显示噬菌体分型可将噬菌体－生物分型的型别进一步划分，ＳＭ６的特性也为霍乱的进一步研究提供了资料。     

Gene structure and genetic localization of the PCLO gene encoding the presynaptic active zone protein Piccolo

Piccolo belongs to a family of presynaptic cytoskeletal proteins likely to be involved in the assembly and function of presynaptic active zones as sites of neurotransmitter release. Given that abnormalities in the formation of synaptic junctions are thought to contribute to cognitive dysfunction during brain development, we have analyzed and compared the gene structure of the Piccolo gene, PCLO, from humans and mice and determined their chromosomal localization. A comparison of the deduced amino acid sequence of cDNA clones encoding Piccolo from human, mouse, rat and chicken reveals the presence of distinct homology domains. Only subsets of these are also present in the structurally related active zone protein Bassoon indicating that Piccolo and Bassoon perform related but distinct functions at active zones. Characterization of the PCLO gene reveals the presence of 25 coding exons spread over 380kb of genomic DNA. The human PCLO gene maps to 7q11.23-q21.3, a region of chromosome 7 implicated as a linkage site for autism and Williams Syndrome suggesting that alterations in the expression of Piccolo or the PCLO gene could contribute to developmental disabilities and mental retardation.     

缺失型DMD／BMD患者PCR快速检测的可行性与意义

应用对应于Ｄｙｓｔｒｏｐｈｉｎ基因缺失热区的二对ＰＣＲ引物和一对内对照无关引物，在同一反应体系中扩增，检测６６例ＤＭＤ／ＢＭＤ患者。发现其中２５例存在１７号或４９号外显子缺失，与同时采用ｃＤＮＡ探针杂交检测出的３５例基因缺失相比．检出率达７１．４％。说明该扩增系统能够作为快速筛查缺失型ＤＭＤ／ＢＭＤ患者的有效手段。这对指导合理选用探针，尤其在产前诊断方面，具有重要意义。