卒中型高血压大鼠心肌线粒体Ca~（2＋），Mg~（2＋）—ATPase及膜流动

本文比较了卒中型自发往高血压鼠（ＳＨＲｓｐ）和京都Ｗｉｓｔａｒ正常血压鼠（ＷＫＹ）心肌线粒体Ｃａ￣（２＋），Ｍｇ￣（２＋）－ＡＴＰａｓｅ活力及膜流动性。用定磷法测得ＷＫＹ和ＳＨＲｓｐ的Ｃａ￣（２＋），Ｍｇ￣（２＋）－ＡＴＰａｓｅ活力（２５℃）分别为０．２８７±０．０１６及０．２１８±０．０１７μｍｏｌ／（ｍｉｎ．ｍｇ）（－ｘ±Ｓｘ）。ＳＨＲｓｐ心肌线粒体Ｃａ￣（２＋），Ｍｇ￣（２＋）－ＡＴＰａｓｅ活力降低２４．７％，两组比较有显著差异（Ｐ＜０．０１，ｎ＝９）．以ＤＰＨ标记心肌线粒体膜，测得ＷＫＹ和ＳＨＲｓｐ的荧光偏振值分别为０．１８７±０．００３及０．１８１±０．００３（Ｐ＞０．０５，ｎ＝６）。     

4种癌基因mRNA在胃癌组织中的表达及意义

应用原位分子杂交（ＩＳＨ）技术，检测８８例胃癌组织中Ｋ－ｒａｓ、Ｈ－ｒａｓ、Ｃ－ｍｙｃ和肿瘤转移抑制基因ｎｍ２３（Ｈ１）ｍＲＮＡ．其阳性结果分别为７８．４％、７０％、５８％和３８．６％，胃癌癌旁移行区正常粘膜和正常胃粘膜上皮的阳性率分别为１８．２％、１７％、１９．３％、２１．６％和０、０、０、３／５，与肿瘤区相比较，除ｎｍ２３外差异均非常显著（Ｐ＜０．０１）。Ｋ－ｒａｓ、Ｈ－ｒａｓｍＲＮＡ表达在在细胞膜内侧，Ｃ－ｍｙｃｍＲＮＡ主要表达在细胞核内，ｎｍ２３－Ｈ１ｍＲＮＡ表达在胞浆内。４种癌基因表达与组织学类型无关，ｎｍ２３－Ｈ１的表达与胃癌病人有无淋巴结转移有关（Ｐ＜０．０１）。     

ATP敏感钾通道对决定小鼠胰岛β－细胞簇放电间期的可能作用

ＡＴＰ敏感钾通道对决定小鼠胰岛β－细胞簇放电间期的可能作用ＷＯＤｉｎｇ，ｅｔａｌ．ＢｌｏｃｍｍｉｃａｅｔＢｉｏｐｈｐｓｉｃａＡｃｔａＢｉｏｍｅｍｂｒａｎｅｓ．１９９６；１２７９（２）２１９～２２６在１０ｍＭ葡萄糖中，翎Ｐ细胞电活动由埃放电与静息期见成...     

用聚合酶链反应—银染单链构象多态性鉴别腮腺炎病毒株

用腮腺炎病毒小疏水蛋白（ＳＨ）基因及其旁侧区的逆转录（ＲＴ）套式聚合酶链反应（Ｎ－ＰＣＲ）产物进行银染单链构象多态性（ＳＳＣＰ）分析，建立了较适的样品变性和聚丙烯酰胺凝胶电泳（ＰＡＧＥ）条件。根据单链电泳模式可将所测６株腮腺炎病毒分为四种ＳＳＣＰ模式，３株兰州野毒株Ｗｌｚ１，Ｗｌｚ２，Ｗｌｚ３呈现一种模式，这３株病毒的ＳＨ基因及其旁侧区ｃＤＮＡ序列之间平均差异仅为０．９６％；两株上海野毒株Ｗｓｈ１和Ｗｓｈ２株的相应序列之间差异为４％，呈现另两种模式。Ｅｎｄｅｒｓ株呈现一种与野毒株差别较大的特殊模式。３株兰州野毒与２株上海野毒相应区域序列之间差异为３．４％～４．５％，故本法可区别ＳＨ基因及其旁侧区ｃＤＮＡ序列之间差异为３．４％的野毒株，且ＳＳＣＰ模式的相近程度与相应区域中核苷酸序列同源性大小一致。此法可用于腮腺炎病毒株分子特性和遣传变异的初步鉴定和分子流行病学研究     

中国4株丁型肝炎病毒抗原编码区基因的cDNA克隆与…

从我国四川、广西、河南的丁型肝炎、病毒抗原（ＨＤＡｇ）或抗体阳性的ＨｂｓＡｇ携带者中，筛选出４株ＨＤＶ ＲＮＡ阳性标本，经逆转录和聚合酶反应（ＲＴ－ＰＣＲ）后，获得包含ＨＤＶ抗原编码区的ｃＤＮＡ片段，并对其进行克隆与序列测定。经计算机分析比较表明：四川、广西株与美国－１株同源性最高，分别为９９．３％、９９．０％；而河南－１、河南－２株与台湾株的同源性较高，分别为９４．３％、９２．１％；上述４株与日     

一株适应MRC－5细胞的减毒HAV的基因测定

一株适应ＭＲＣ－５细胞的减毒ＨＡＶ的基因测定［英］／ＦｕｎｋｈｏｕｓｅｒＡＷ…ＪＶｉ－ｒｏｌ．－１９９４，６８（１）．１４８～１５７这是一株ＨＡＶ活疫苗的候选株，是适应原代非洲绿猴肾（ＡＧＭＫ）细胞的ＨＭ－１７５株，再使此株ＨＡＶ适应于ＭＲＣ－５细胞...     

应用ED－PCR技术快速检测耐甲氧西林葡萄球菌

应用ＥＤ－ＰＣＲ（Ｅｎｚｙｍａｔｉｃｄｅｔｅｃｔｉｏｎｏｆｐｏｌｙｍｅｒａｓｅ－ｃｈａｉｎｒｅａｃｔｉｏｎ）技术从临床分离的７６株葡萄球菌中鉴定出耐甲氧西林金黄色葡萄球菌（Ｍｅｔｈｉｃｉｌｉｎ－ｒｅｓｉｓｔａｎｔｓｔａｐｈｙｌｏｃｏｃｃｕｓａｕｒｅｕｓ，ＭＲＳＡ）９株，耐甲氧西林血浆凝固酶阴性的葡萄球菌（Ｍｅｔｈｉｃｉｌｉｎ－ｒｅｓｉｓｔａｎｔｃｏａｇｅｌａｓｅ－ｎｅｇａｔｉｖｅｓｔａｐｈｙｌｏｃｏｃｃｉ，ＭＲ－ＣＮＳ）１１株。此结果与药物敏感性检测结果相符。ＥＤ－ＰＣＲ法具有简便、特异、省时，耗资少等优点，宜在临床检验室推广应用。     

慢性肝炎和肝癌病人血清中乙型肝炎病毒DNA的检测

为了了解慢性肝炎和肝癌病人患者体内乙型肝炎病毒（ＨＢＶ）复制与ＨＢＶ血清标志之间的关系，用酶联免疫吸附实验（ＥＬＩＳＡ）、聚合酶链反应（ＰＣＲ）及斑点杂交方法对６１例慢性肝炎和４７例肝癌患者的ＨＢＶ表面抗原（ＨＢｓＡｇ）、相关ｅ抗原（ＨＢｅＡｇ）、表面抗体（抗－ＨＢｓ）、核心抗体（抗－ＨＢｃ）、相关ｅ抗体（抗－ＨＢｅ）进行了检测。结果表明：ＨＢＶＤＮＡ在ＨＢｓＡｇ、ＨＢｅＡｇ、／抗－ＨＢｃ阳性的慢性肝炎和肝癌患者血清中的检出率分别为９０．５０％和５０．００％；在ＨＢｓＡｇ／抗－ＨＢｅ、抗－ＨＢｃ阳性者的检出率分别为４５．４０％和７．１４％；在ＨＢｓＡｇ阳性、ＨＢｅＡｇ阴性／抗－ＨＢｅ阴性者中的检出率分别为６０．００％和４０．００％；ＨＢｓＡｇ阴性、／抗－ＨＢｃ阳性或／抗－ＨＢｅ阳性或／抗－ＨＢｓ阳性者中的检出率分别为２０．００％和２２．２２％；在血清学指标全阴性时，慢性肝炎和肝癌患者血清中ＨＢＶＤＮＡ的检出率均为０。实验提示：无论是肝炎或肝癌，在ＨＢｓＡｇ、ＨＢｅＡｇ同时阳性时，ＨＢＶ复制最为活跃；在单独ＨＢｓＡｇ阳性时，ＨＢＶ有一定程度的复制；ＨＢＶ复制在肝癌细胞中受到一定程度的抑制。     

TNF相关的凋亡诱导配体及其受体与凋亡

ＴＮＦ相关的凋亡诱导配体（ＴＮＦ－ｒｅｌａｔｅｄ ａｐｏｐｔｏｓｉｓ－ｉｎｄｕｃｉｎｇ ｌｉｇａｎｄ，ＴＲＡＩＬ）是新发现的ＴＮＦ超家族成员，与新近发现的受体（ＴＮＦ－ｒｅｌａｔｅｄ ａｐｏｐｔｏｓｉｓ－ｉｎｄｕｃｉｎｇ ｌｉｇａｎｄ ｒｅｃｅｐｔｏｒ，ＴＲＡＩＬＲ）相结合的启动细胞内的信号转导，在体外迅速诱导各种来源的细胞株的凋亡，本文综述了ＴＲＡＩＬ和ＴＲＡＩＬＲ结构功能的分子基础及其诱导凋亡     

RADS抑郁量表的修订

本文报导应用ＲｅｙｎｏｌｄｓＡｄｏｌｅｓｃｅｎｔＤｅｐｒｅｓｓｉｏｎｓｃａｌｅ（ＲＡＤＳ）抑郁量表在全国７省市对３９０５名中学生测量的结果，量表重测相关系数为０．７９、Ｃｒｏｎｂｅｃｈα系数为０．８８、分半信度相关系数为０．８８；与流调中心用抑郁量表（ＣＥＳ－Ｄ）的相关系数为０．８４；因子分析结果表明该量表有较好的结构效度。     

Identification by Subtractive Hybridization of a Novel Insertion Sequence Specific for Virulent Strains of Porphyromonas gingivalis

Subtractive hybridization was employed to isolate specific genes from virulent Porphyromonas gingivalis strains that are possibly related to abscess formation. The genomic DNA from the virulent strain P. gingivalis W83 was subtracted with DNA from the avirulent strain ATCC 33277. Three clones unique to strain W83 were isolated and sequenced. The cloned DNA fragments were 885, 369, and 132 bp and had slight homology with only Bacillus stearothermophilus IS5377, which is a putative transposase. The regions flanking the cloned DNA fragments were isolated and sequenced, and the gene structure around the clones was revealed. These three clones were located side-by-side in a gene reported as an outer membrane protein. The three clones interrupt the open reading frame of the outer membrane protein gene. This inserted DNA, consisting of three isolated clones, was designated IS1598, which was 1,396 bp (i.e., a 1,158-bp open reading frame) in length and was flanked by 16-bp terminal inverted repeats and a 9-bp duplicated target sequence. IS1598 was detected in P. gingivalis W83, W50, and FDC 381 by Southern hybridization. All three P. gingivalis strains have been shown to possess abscess-forming ability in animal models. However, IS1598 was not detected in avirulent strains of P. gingivalis, including ATCC 33277. The IS1598 may interrupt the synthesis of the outer membrane protein, resulting in changes in the structure of the bacterial outer membrane. The IS1598 isolated in this study is a novel insertion element which might be a specific marker for virulent P. gingivalis strains.     

PCR Analyses of tRNA Intergenic Spacer, 16S-23S Internal Transcribed Spacer, and Randomly Amplified Polymorphic DNA Reveal Inter- and Intraspecific Relationships of Enterobacter cloacae Strains

PCR analysis of tRNA intergenic spacer (tDNA-PCR) and of the 16S-23S internal transcribed spacer (ITS-PCR) and random amplified polymorphic DNA (RAPD) analysis were evaluated for their usefulness in characterization of Enterobacter cloacae strains isolated from both clinical origins and vaccine microbial contamination. tDNA-PCR presented specific and reproducible patterns for Enterobacter sakazakii ATCC 29004, Enterobacter aerogenes ATCC 13048, and Enterobacter cloacae ATCC 13047 and 23355 that presented the same profile for all 16 E. cloacae isolates, offering an alternative tool for species-level identification. ITS-PCR and RAPD analysis yielded completely different banding patterns for the 20 strains studied, except for E. cloacae strains isolated from different batches of vaccine that exhibited a unique pattern, suggesting contamination by the same strain. The combined use of tDNA-PCR and ITS-PCR in a one-step protocol allows accurate identification and typing of E. cloacae strains a few hours after the colony has been isolated.     

Comparison of four DNA-based methods for strain delineation of Candida lusitaniae.

Four methods for the accurate delineation of epidemiologically related and unrelated strains of Candida lusitaniae were compared. Three pulsed-field electrophoretic methods, including two contour-clamped homogeneous field gel electrophoresis methods (EKP-1 and EKP-2) yielding electrophoretic karyotype patterns of intact chromosomal DNA and a method in which the chromosomal DNA was macrodigested with the endonuclease SfiI prior to pulsed-field electrophoresis (MDP), and a random amplified polymorphic DNA (RAPD) assay were evaluated. A selected panel of 21 well-characterized isolated representing 13 strains of C. lusitaniae, including 7 epidemiologically related isolates of one strain (group I-A), 3 epidemiologically related isolates of another strain (group I-B), and 11 epidemiologically unrelated isolates (group II), were tested. All isolates were coded and tested in a blinded manner. All seven group I-A isolates were confirmed to be a single strain by the EKP-1 and MDP methods, and the three group I-B isolates were shown to be a single strain by the EKP-1, EKP-2, MDP, and RAPD methods. Subtle differences were noted with two of the group I-A isolates by the EKP-2 method, whereas three of these isolates were different by the RAPD method. Each group II isolate had distinct patterns by all four methods. These data support the fact that the three pulsed-field electrophoretic methods and the RAPD method can be used to delineate strains of C. lusitaniae.(ABSTRACT TRUNCATED AT 250 WORDS)     

Genomic Loci of the Porphyromonas gingivalis Insertion Element IS1126

The Porphyromonas gingivalis genome contains multiple copies of insertion element IS1126. When chromosomal DNA digests of different strains were probed with IS1126, between 25 and 35 hybridizing fragments per genome were detected, depending on the strain. Unrelated strains had very different restriction fragment length polymorphism (RFLP) patterns. When different laboratory copies of a specific strain were examined, the IS1126 RFLP patterns were very similar but small differences were observed, indicating that element-associated changes had occurred during laboratory passage. Within the next year, genome sequencing, assembly, and annotation for P. gingivalis W83 will be completed. Because repetitive elements complicate the assembly of randomly sequenced DNA fragments, we isolated and sequenced the flanking regions of IS1126 copies in strain W83. We also isolated and sequenced the flanking regions of IS1126 copies in strain ATCC 33277 in order to compare insertion sites in phylogenetically divergent strains. We identified 37 new sequences flanking IS1126 from strain ATCC 33277 and 30 from strain W83. The insertion element was found between genes except where it transposed into another insertion element. Examination of identifiable flanking genes or open reading frames indicated that the insertion sites were different in the two strains, except that both strains possess an insertion adjacent to the Lys-gingipain gene (J. P. Lewis and F. L. Macrina, Infect. Immun. 66:3035-3042, 1998). Most of the genes or sequences flanking IS1126 in ATCC 33277 were present in W83 but were contiguous and not insertion element associated. Thus, where genes were identified in both strains, their order was maintained, indicating that the two genomes are organized similarly, but the loci of IS1126 are different. In both strains, insertion element-associated duplicated target sites were lost from several copies of IS1126, providing evidence of homologous recombination between elements. Larger organizational differences between the genomes, such as deletions and inversions, may result from insertion element-mediated recombination events.     

Blastogenic response of human lymphocytes to oral bacterial antigens: characterization of bacterial sonicates.

Soluble sonicate supernatant preparations were made from Actinomyces viscosus (ATCC 19246), A. naeslundii (ATCC 12104), two strains of Veillonella alcalescens (strain HV-1 and a human oral isolate), Streptococcus sanguis (ATCC 10556), S. mutans (strain 6715-T2), Bacteroides melaninogenicus (strain K110), and Leptotrichia buccalis (isolated from human dental plaque). These supernatants were characterized with reference to their chemical and antigenic components and their biological activity determined by using in vitro lymphocyte blastogenesis as a measure of the host's cellular immune response. The sonicate supernatant of each bacterium contained protein, neutral sugars, methylpentose, and nucleic acids. Protein was the major component in all except L. buccalis, in which neutral sugars predominated. The antigenic components in each supernatant were detected by using rabbit antisera prepared against the whole bacteria and the sonicate supernatant. The supernatants showed a complex antigenic distribution on immunoelectrophoretic analysis. The supernatants were shown to be antigenic and not mitogenic in nature, since neither cord blood lymphocytes nor all adult lymphocytes were stimulated. The supernatant antigen preparations showed a reproducible, dose-dependent, and kinetic response in vitro, which was similar to that seen with the antigen preparation streptokinase-streptodornase.     

A highly specific and sensitive DNA probe derived from chromosomal DNA of Helicobacter pylori is useful for typing H. pylori isolates.

HindIII-digested DNA fragments derived from an EcoRI-digested 6.5-kb fragment of chromosomal DNA prepared from Helicobacter pylori ATCC 43629 (type strain) were cloned into the pUC19 vector. A 0.86-kb insert was identified as a potential chromosomal DNA probe. The specificity of the probe was evaluated by testing 166 non-H. pylori bacterial strains representing 38 genera and 91 species which included aerobic, anaerobic, and microaerophilic flora of the upper and lower gastrointestinal tracts. None of the 166 non-H. pylori strains hybridized with this probe (100% specificity), and the sensitivity of this probe was also 100% when H. pylori isolates from 72 patients with gastritis and with the homologous ATCC type strain were tested by dot blot hybridization. The capability of this probe for differentiating between strains of H. pylori was evaluated by Southern blot hybridization of HaeIII-digested chromosomal DNA from 68 clinical isolates and the homologous ATCC type strain of H. pylori. Fifty-one unique hybridization patterns were seen among the 69 strains tested, demonstrating considerable genotypic variation among H. pylori clinical isolates. We propose that this probe would be of significant value for conducting epidemiologic studies.     

Legionella cincinnatiensis sp. nov. isolated from a patient with pneumonia.

A Legionella-like organism (strain 72-OH-H [= ATCC 43753]) was isolated from an open-lung biopsy specimen from a hemodialysis patient with end-stage renal disease and bronchopneumonia. Growth characteristics and gas-liquid chromatographic profiles of the isolate were consistent with those for Legionella spp. The isolate was presumptively identified as a Legionella longbeachae serogroup 1 strain by direct immunofluorescence staining. However, the organism was serologically distinct in the slide agglutination test with absorbed antisera. DNA hybridization studies showed that strain 72-OH-H constitutes a new Legionella species, which is named Legionella cincinnatiensis (ATCC 43753).     

Efficient discrimination of Mycobacterium tuberculosis strains by 16S-23S spacer region-based random amplified polymorphic DNA analysis.

Amplification of the region separating the genes coding for 16S and 23S rRNA was performed with 15 Mycobacterium tuberculosis isolates and the type strain, ATCC 27294. Reproducible amplification patterns were obtained. PCR products were then used as target DNA for random amplified polymorphic DNA (RAPD) analysis. The discriminatory power was higher than when whole genomic DNA was used as a RAPD template. 16S-23S spacer region-based RAPD analysis was a simple and efficient method of differentiation. Consequently, it may be a useful tool for epidemiologic studies of tuberculosis.     

DNA-DNA hybridization incompatibility of Campylobacter pylori with other Campylobacter and Wolinella species

DNA-DNA hybridization in solution was used to characterize 23 human isolates of Campylobacter pylori. The 23 isolates showed DNA affinity with the type strain (NCTC 11637). The relative binding ratios varied between 0.83 and 1. Type strains of C. coli (NCTC 11366), C. jejuni (NCTC 11351), C. laridis (NCTC 11352), C. sputorum subsp. sputorum (ATCC 35980), Wolinella recta (NCTC 11489) and W. succinogenes (ATCC 29543) showed relative binding ratios less than 0.01 compared to the C. pylori type strain. The results suggest that C. pylori is a homogenous taxonomic unit distinctly separated from these other species.     

铜绿假单胞菌随机扩增多态性DNA指纹法基因分型研究

建立了铜绿假单胞菌（ＰＡ）随机扩增多态性ＤＮＡ（ＲＡＰＤ）指纹图基因分型方法，并应用于流行病学相关或不相关的７５个ＰＡ菌株的基因分型。分型率为１００％。３２株新生儿暴发感染菌株，可分成４个血清型和５个ＲＡＰＤ谱型，其中２５株属于同一谱型。在８例患者的垂直追踪菌株中，除有１例其第１和第３个分离株的血清型和ＲＡＰＤ谱型均相同，但与第２个分离株的血清型和ＲＡＰＤ谱型不同外，其余７例前后菌株的谱型相同。１３株散发菌株可分成３个血清型和１０个ＲＡＰＤ谱型；１２株流行病学无关菌株的ＲＡＰＤ谱型均不相同。本研究结果表明，ＲＡＰＤ指纹图基因分型法具有分型率高、分辨力强、比较快速简便等优点，并且不需要已知核酸序列，是在分子水平上对微生物感染的病原学、发病机理及流行病学研究的较理想的方法。