Molecular epidemiology of Vibrio cholerae O139 in China: polymorphism of ribotypes and CTX elements

Vibrio cholerae O139, the second etiological serogroup of cholera, triggered the first outbreak of O139 cholera in China in 1993. To analyze the clone polymorphism of O139 isolates in China, 117 strains of V. cholerae O139, isolated from different areas in China between 1993 and 1999, were selected to characterize the phylogenetic relationships by molecular techniques. Analysis of restriction fragment length polymorphism in the conserved 16S rRNA gene revealed seven different ribotypes within the 117 strains. Among these strains, there were eight that lacked the cholera toxin gene (ctxAB), zot, and the repetitive sequence (RS); these eight strains belonged to three individual ribotypes. Our results suggested that V. cholerae O139 strains in China had clone diversity in phylogeny. The results of our hybridization patterns for CTX genetic elements (ctxAB, zot, and RS) showed that CTXPhi genomes in most V. cholerae O139 strains had two or more copies and had extensive restriction patterns even for the strains which belong to the same ribotype. For 22 (20.1%) strains, the copies of ctxAB were different from those of zot, suggesting that a ctxAB-negative CTXPhi genome may exist in O139 strains. This ctxAB-negative CTXPhi genome may coexist with the intact CTXPhi genome in a strain. In addition, the dendrogram for I-CeuI-generated pulsed-field gel electrophoresis patterns showed that V. cholerae serogroup O139 has a closer relationship with one strain of serogroup O22 than with the strains of serogroup O1. The results of this study showed the clonal diversity and the distribution of O139 strains in China, suggesting multiple origins of the O139 cholera epidemic or sporadic events.     

RINm5f cells express inactivating BK channels whereas HIT cells express noninactivating BK channels

Large-conductance Ca2+- and voltage-activated BK-type K+ channels are expressed abundantly in normal rat pancreatic islet cells and in the clonal rat insulinoma tumor (RINm5f) and hamster insulinoma tumor (HIT) beta cell lines. Previous work has suggested that the Ca2+ sensitivity of BK channels in RIN cells is substantially less than that in HIT cells, perhaps contributing to differences between the cell lines in responsiveness to glucose in mediating insulin secretion. In both RIN cells and normal pancreatic beta cells, BK channels are thought to play a limited role in responses of beta cells to secretagogues and in the electrical activity of beta cells. Here we examine in detail the properties of BK channels in RIN and HIT cells using inside-out patches and whole cell recordings. BK channels in RIN cells exhibit rapid inactivation that results in an anomalous steady-state Ca2+ dependence of activation. In contrast, BK channels in HIT cells exhibit the more usual noninactivating behavior. When BK inactivation is taken into account, the Ca2+ and voltage dependence of activation of BK channels in RIN and HIT cells is essentially indistinguishable. The properties of BK channel inactivation in RIN cells are similar to those of inactivating BK channels (termed BKi channels) previously identified in rat chromaffin cells. Inactivation involves multiple, trypsin-sensitive cytosolic domains and exhibits a dependence on Ca2+ and voltage that appears to arise from coupling to channel activation. In addition, the rates of inactivation onset and recovery are similar to that of BKi channels in chromaffin cells. The charybdotoxin (CTX) sensitivity of BKi currents is somewhat less than that of the noninactivating BK variant. Action potential voltage-clamp waveforms indicate that BK current is activated only weakly by Ca2+ influx in RIN cells but more strongly activated in HIT cells even when Ca2+ current magnitude is comparable. Concentrations of CTX sufficient to block BKi current in RIN cells have no effect on action potential activity initiated by glucose or DC injection. Despite its abundant expression in RIN cells, BKi current appears to play little role in action potential activity initiated by glucose or DC injection in RIN cells, but BK current may play an important role in action potential repolarization in HIT cells.     

Energy spectra, angular spread, fluence profiles and dose distributions of 6 and 18 MV photon beams: results of monte carlo simulations for a varian 2100EX accelerator

The purpose of this study is to provide detailed characteristics of incident photon beams for different field sizes and beam energies. This information is critical to the future development of accurate treatment planning systems. It also enhances our knowledge of radiotherapy photon beams. The EGS4 Monte Carlo code, BEAM, has been used to simulate 6 and 18 MV photon beams from a Varian Clinac-2100EX accelerator. A simulated realistic beam is stored in a phase space data file, which contains details of each particle's complete history including where it has been and where it has interacted. The phase space files are analysed to obtain energy spectra, angular distribution, fluence profile and mean energy profiles at the phantom surface for particles separated according to their charge and history. The accuracy of a simulated beam is validated by the excellent agreement between the Monte Carlo calculated and measured dose distributions. Measured depth-dose curves are obtained from depth-ionization curves by accounting for newly introduced chamber fluence corrections and the stopping-power ratios for realistic beams. The study presents calculated depth-dose components from different particles as well as calculated surface dose and contribution from different particles to surface dose across the field. It is shown that the increase of surface dose with the increase of the field size is mainly due to the increase of incident contaminant charged particles. At 6 MV, the incident charged particles contribute 7% to 21% of maximum dose at the surface when the field size increases from 10 x 10 to 40 x 40 cm2. At 18 MV, their contributions are up to 11% and 29% of maximum dose at the surface for 10 x 10 cm2 and 40 x 40 cm2 fields respectively. However, the fluence of these incident charged particles is less than 1% of incident photon fluence in all cases.     

细胞角蛋白19、galectin-3、HBME-1在甲状腺病变上的表达及鉴别诊断意义

目的 研究细胞角蛋白(CK)19、galectin(Gal)-3、HBME-1在甲状腺不同病变表达的特点及鉴别诊断中的应用价值。方法 应用免疫组织化学EnVision法检测了21例结节性甲状腺肿(结甲)、14例毒性甲状腺肿(甲亢)、15例甲状腺滤泡性腺瘤(腺瘤)、13例滤泡性癌、13例滤泡型乳头状癌及48例经典型乳头状癌中单克隆抗体CK19、Gal-3、HBME-1的表达。结果 甲状腺病变中3种标记表达均位于细胞质;CK19、Gal-3、HBME-1的表达在甲状腺良性病变(结甲、甲亢、腺瘤)大多为弱阳性或阴性,而滤泡性癌阳性明显增加、乳头状癌(滤泡型及经典型)大多为中、强阳性,3种标记在甲状腺不同病变的阳性表达率结甲为52．4％(11／21)、9．5％(2／21)、19．0％(4／21),甲亢为50．0％(7／14)、7．1％(1／14)、7．1％(1／14),腺瘤为60％(9／15)、13．3％(2／15)、13．3％(2／15),滤泡性癌为76．9％(10／13)、61．5％(8／13)、53．8％(7／13),滤泡型乳头状癌为：100％(13／13)、84．6％(11／13)、92．3％(12／13),经典型乳头状癌为100％(48／48)、93．8％(45／48)、95．8％(46／48);在甲状腺良性病变(结甲、甲亢、腺瘤)与恶性病变(滤泡性癌、乳头状癌)间3种标记差异均有显著性(P均=0．000);同时3种标记在滤泡样病变即腺瘤、滤泡性癌和滤泡型乳头状癌间亦有显著差异(CK19：P=0．038,Gal-3：P=0．001,HBME-1：P=0．000)。结甲有9例,甲亢有7例,腺瘤有6例3种标记均不表达,滤泡性癌仅有1例,而乳头状癌(滤泡型及经典型)没有病例3种标记均不表达,同一病例有2种以上阳性表达在结甲、甲亢、腺瘤、滤泡性癌、滤泡型乳头状癌和经典型乳头状癌中分别为14．2％(3／21)、21．4％(3／14)、20．0％(3／15)、69．2％(9／13)、92．3％(12／13)、100．0%(48／48),在甲状腺良性病变与恶性病变间以及滤泡样病变间差异亦有显著性(P=0．000)。结论 CK19、Gal-3、HBME-1的检测尤其是联合检测对甲状腺病变的诊断、鉴别诊断具有较高的实用价值。     

大鼠视皮质胼胝体轴突的生后发育和形态──生物素结合的葡聚糖胺顺行示踪法研究

采用生物素结合的葡聚糖胺顺行示踪法研究了大鼠视皮质主要胼胝体投射区即１７／１８ａ交界区胼胝体轴突的生后发育和形态。在生后５天时，此交界区胼胝体轴突从白质向灰质Ⅰ层垂直生长，在灰质内仅有极少量的侧支抽芽。至生后１３天时，皮质Ⅰ层最先出现致密的由胼胝体轴突终支组成的终末丛。到生后１７天时，类似的终末丛也见于皮质Ⅱ／Ⅲ，Ⅴ和Ⅵ层，这种分布型式与成年大鼠者相似。以上结果表明，绝大部分胼胝体轴突首先生长到达Ⅰ层并先在Ⅰ层发出终支，然后再在其它皮质层发出侧支及终支，因而提示皮质Ⅰ层在胼胝体联系的生后发育中可能发挥重要作用。     

用PCR扩增tim基因检测蓝氏贾第鞭毛虫

采用聚合酶链反应 (PCR)对蓝氏贾第鞭毛虫 (Giardialamblia)的磷酸丙糖异构酶 (triosephosphateisomerase ,缩写为tim)基因进行特异性扩增 ,结果扩增出 1条 6 83bp的DNA片段。此方法的特异性可高达10 0 % ,而其它DNA样本 ,如日本血吸虫 (Schistosomajaponicum)、刚地弓形虫 (Toxoplasmagondii)、微小隐孢子虫 (Cryptosporidiumparvum)、溶组织内阿米巴 (Entamoebahistolytica)、旋毛虫 (Trichinellaspiralis)和阴道毛滴虫 (Trichomonasvaginalis) ,以及人体血细胞等均未出现扩增反应。本法的敏感性也很高 ,可检测到0 4pg贾第虫包囊的DNA。 13株来自不同地理位置和 或宿主的贾第虫DNA样本在PCR中均各产生 1条长为 6 83bp的目的片段。上述结果表明本实验建立的检测贾第虫的PCR方法有效     

心理健康对精神分裂症患者认知功能的影响

目的观察心理健康对精神分裂症患者认知功能的影响。方法采用入院顺序分层随机法，将120例慢性精神分裂症患者分为研究组（心理健康＋奋乃静）和对照组（单用奋乃静）治疗。在治疗前，治疗后4、8、12周末分别以阳性症状和阴性症状量表（PNASS？评定疗效，用韦氏成人智力量表（WAIS—R）、韦氏记忆量表（WMS）和威斯康星卡片分类测验（wCST）评定治疗前后患者认知功能的改变。结果治疗12周后，研究组的言语量表、操作量表、全量表和记忆量表分分别为84．97±3．73、80．41±5．93、83．37±5．81、77．26±13．31；对照组分别为75．41±3．95、72．43±5．56、75．87±5．34、72．15±13．19，两组有显著差异（P〈0．01）。PANSS总分、阴性因子分比治疗前明显降低。结论心理健康治疗对精神分裂症患者认知功能有明显改善。     

Identification of novel regions of allelic loss from a genomewide scan of esophageal squamous-cell carcinoma in a high-risk Chinese population

Esophageal cancer is one of the most common fatal cancers worldwide. Deletions of genomic regions are thought to be important in esophageal carcinogenesis. We conducted a genomewide scan for regions of allelic loss using microdissected DNA from 11 esophageal squamous-cell carcinoma patients with a family history of upper gastrointestinal tract cancer from a high-risk region in north central China. Allelic patterns of 366 fluorescently labeled microsatellite markers distributed at 10-cM intervals over the 22 autosomal chromosomes were examined. We identified 14 regions with very high frequency (>/= 75%) loss of heterozygosity (LOH), including broad regions encompassing whole chromosome arms (on 3p, 5q, 9p, 9q, and 13q), regions of intermediate size (on 2q, 4p, 11p, and 15q), and more discrete regions identified by very high frequency LOH for a single marker (on 4q, 6q, 8p, 14q, and 17p). Among these 14 regions were 7 not previously described in esophageal squamous-cell carcinoma as having very high frequency LOH (on 2q, 4p, 4q, 6q, 8p, 14q, and 15q). The very high frequency LOH regions identified here may point to major susceptibility genes, including potential tumor suppressor genes and inherited gene loci, which will assist in understanding the molecular events involved in esophageal carcinogenesis and may help in the development of markers for genetic susceptibility testing and screening for the early detection of this cancer. Genes Chromosomes Cancer 27:217-228, 2000. Published 2000 Wiley-Liss, Inc.     

结核性淋巴结炎的组织细胞反应性增生变型

应用结核杆菌ＤＮＡ１２３ｂｐ特异性序列片段为靶序列的多聚酶链反应（Ｍ·ＴＢ－ＰＣＲ）技术、ＢＣＧ免疫组化（ＢＣＧ－ＩＨＣ）技术和抗酸染色（ＡＦ）方法，对３８例呈现组织细胞反应性增生－碎屑样坏死－嗜中性白细胞渗出病变的淋巴结石蜡包埋组织进行了分支杆菌／结核杆菌的回顾性检测。三种方法的综合阳性率为５２．６％（２０／３８例）。ＡＦ、ＢＣＧ－ＩＨＣ和Ｍ·ＴＢ－ＰＣＲ的各自阳性率分别为０．８％、２６．３％和５０％。研究结果表明：（１）在按本文标准选择的淋巴结“组织细胞反应性增生”病变中，有半数病例与结核杆菌的感染有关，即结核性淋巴结炎可呈现“组织细胞反应性增生”之类的变型；（２）ＰＣＲ技术在结核性淋巴结炎的病原学诊断上具有重要价值。     

猫阴部神经传入传出成分在脊髓内的定位分布—HRP逆行及跨越神经节追踪研究

向猫阴部神经及其分支注入HRP溶液,观察并分析了逆行和跨节追踪的结果。 1.证实阴部神经为含躯体传出、传入,内脏传出、传入纤维的混合神经,其内脏性传出、传入成分仅存在于阴茎背神经中。 2.逆行标记细胞出现于L_7—S_3前角Onuf核和S_(1-3)的中间带外侧核。本文结果表明,Onuf核是支配盆底横纹肌的运动核,但它和其它前角运动核在细胞形态和活性物质的分布上都明显不同。特别是它的一部分神经元的树突形成树突束到达中间带外侧核。本文结合排尿、排便功能从形态学上较详细地讨论了Onuf核和中间带外侧核的关系。 3.本文证明阴部神经领域的内脏初级传入(来自阴茎背神经)和躯体初级传入纤维都向骶髓后连合核区有浓密的投射。结合以往的工作讨论了盆腔脏器、外生殖器的内脏传入和坐骨神经、阴部神经的躯体传入在骶髓后连合核区汇聚的现象及机能意义,推测这种汇聚可能是产生牵涉痛和针刺镇痛机制的形态学基础。