小儿肺炎时血液流变学的变化

本实验对１７例小儿肺炎患者进行了血液流变学指标的检测。结果表明：肺炎组高切变率（２ｏｏｓ－１）下校正全血粘度（ｃｏｒｒｅｃｔｗｈｏｌｅｂｌｏｏｄｖｉｓｃｏｓｉｔｙ，ＣＷＢＶ）和相对全血粘度（ｒｅｌａｔｉｖｅｗｈｏｌｅｂｌｏｏｄｖｉｓｃｏｓｉｔｙ，ＲＷＢＶ）均明显高于正常对照组；低切变率（１０ｓ￣（－１））下ＣＷＢＶ和ＲＷＢＶ也高于正常对照组，校正ＥＳＲ值下降，血沉方程Ｋ值减小，纤维蛋白原（ｆｉｂｒｉｎｏｇｅｎ，Ｆｇ）含量与高、低切变率下ＣＷＢＶ和ＲＷＢＶ显著相关，与血浆粘度（ηｐ）也显著相关；红细胞聚集指数（ｒｅｄｃｅｌｌａｇｇｒｅｇａｔｉｏｎｉｎ－ｄｅｘ，ＲＡＩ）与高、低切变率下ＣＷＢＶ也显著相关。提示小儿肺炎时全血及血浆粘度均增高，红细胞刚性增加，聚集性增强。     

对家兔两种心肌梗塞模型的观察

结扎家兔冠状动脉前降支或左室支复制心肌梗塞的动物模型,观察血液动力学和心电图变化,6小时后取出心脏进行TTC染色,测定心肌梗塞范围,并作病理切片。结果表明,结扎前降支复制心肌梗塞模型,阳性率较低,梗塞范围小,部位不固定,血液动力学、心电图和镜下改变均不如结扎左室支明显。     

缺血预处理对兔缺血心肌的保护作用

采用动物缺血－再灌注模型，结扎兔冠状动脉左前降支（LAD）造成急性心肌梗塞（AMI）。实验组经过5min缺血，10min再灌注后持续缺血60min，3h再灌注；对照组直接造成60min缺血，3h再灌注。心肌梗塞范围由1％三苯四氮唑兰（TTC）染色确定，并以梗塞范围占缺血范围重量的百分比表示。结果：实验组梗死心肌明显少于对照组（P＜0.01）；实验组发生室性心律失常率明显低于对照组（P＜0．05）；实验组再灌注后血小板表面α－颗粒膜蛋白（GMP－140）分子数明显少于对照组（P＜0．05）。表明：预适应（Preconditioning，PC）具有心肌保护作用。     

用MCV／RDW对地中海贫血初筛探讨

本文采用美国ＣＤ３５００型细胞分析仪，对４９例地中海贫血患者、２０例缺铁性贫血患者和１０４例健康人进行了红细胞平均体积（ＭＣＶ）／红细胞体积分布宽度（ＲＤＷ）检测。结果表明１０４例健康人ＭＣＶ／ＲＤＷ均在正常参考值范围，２０例缺铁性贫血为小细胞不均一性贫血（ＭＣＶ↓／ＲＤＷ↑），与文献报道相符。２４例重型和中间型β地中海贫血和１０例血红蛋白Ｈ病患者亦表现为小细胞不均一性，且ＲＤＷ值的改变较缺铁性贫血更显著（Ｐ＜０．０１）。１５例轻型β地中海贫血ＭＣＶ／ＲＤＷ较健康人轻度增高，其ＭＣＶ／ＲＤＷ值的改变与缺铁性贫血无显著差异（Ｐ＞０．０５），地中海贫血患者的贫血程度与ＲＤＷ值的增高呈平行的关系     

β－半乳糖苷酶基因的腺伴随病毒（AAV－2）载体的构建及其在哺乳类细胞中的表达

以野生型２型人类腺伴随病毒（ＡＡＶ－２）全基因组载体（ｐＳＳＶ９和ｐＳＳＶ９－ｉｎｔ－）为基础，构建了重组ＡＡＶ载体ｐＳＳＶ９／Ｐ４０－ＬａｃＺ（＋）和ｐＳＳＶ９／Ｐ４０－ＬａｃＺ（－），制备ＡＡＶ重组病毒，感染宿主细胞后表达了β－半乳糖苷酶基因。此外，用脂质体转染法将载体导入２９３细胞和Ａ５４９细胞，进行了ＬａｃＺ基因瞬时表达研究。结果显示，重组ＡＡＶ载体（ＰＳＳＶ９／Ｐ４０－ＬａｃＺ（＋））ＬａｃＺ基因表达的动态变化特点是，转染后表达较早（１２～２４ｈ）出现，并迅速达到高峰值（经２４ｈ），随后较快下降（经４８～７２ｈ）并维持在较低水平。将启动子附近ｉｔｒ结构有差别的上述两载体转染宿主细胞７２ｈ后做表达水平比较，结果ｐＳＳＶ９／Ｐ４０－ＬａｃＺ（－）在２９３细胞和Ａ５４９细胞中，ＬａｃＺ表达水平均高于ｐＳＳＶ９／Ｐ４０－ＬａｃＺ（＋）（１．２倍～１．４倍），提示启动子附近ＡＡＶｉｔｒ结构的变异对表达有一定的影响。     

大鼠实验性左室心肌梗塞对右室收缩性能的影响

对左室心肌梗塞(MI)是否会直接影响右室功能,至今尚有争议。本实验观察大鼠左室MI时右室dp/dt max的改变及其与左室dp/dt max和梗塞范围(IS)间的关系。结果发现冠脉结扎后1天,在左室dp/dt max显著降低的同时,右室dp/dt max也显著降低,且与左室dp/dt max呈显著的直线正相关,而与IS呈显著的直线负相关。在冠脉结扎后3天时,左室dp/dt max有显著恢复,但仍低于对照水平,而右室dp/dt max已恢复正常,且与左室的dp/dt max和IS间不再具有显著的直线相关关系。由此证明;在大鼠左室MI早期,右室收缩性能可受到直接影响,影响的程度与IS及左室收缩性能降低的程度相关;但当左室收缩性能恢复到一定程度时,这种影响消失。     

β—半乳糖苷酶基因的腺伴随病毒（AAV—2）载体的构建及其在…

以野生型２型人类腺伴随病毒（ＡＡＶ－２）全基因组载体（ｐＳＳＶ９和ｐＳＳＶ９－ｉｎｔ＾－）为基础，构建了重组ＡＡＶ载体ｐＳＳＶ９／Ｐ４０－ＬａｃＺ（＋）和ｐＳＳＶ９／Ｐ４０－ＬａｃＺ（－），制备ＡＡＶ重组病毒，感染宿主细胞后表达了β－半乳糖苷酶基因。此外，用脂质体转染法将载体导入２９３细胞和Ａ５４９细胞，进行了ＬａｃＺ基因瞬时表达研究。结果显示，重组ＡＡＶ载体（ｐＳＳＶ９／Ｐ４０－ＬａｃＺ（＋））     

ER阳性和ER阴性人乳腺癌细胞株p53、mdm-2及p21~(WAF1)蛋白表达的研究

目的研究ＥＲ阳性和ＥＲ阴性人乳腺癌细胞株ｐ５３、ｍｄｍ－２和ｐ２１ＷＡＦ１蛋白的表达及其与细胞生物学特性的关系。方法应用细胞培养、基因转染和免疫组化染色ＬＳＡＢ法等技术，检测ＥＲ阳性、表达野生型ｐ５３（ｗｔｐ５３）蛋白的ＭＣＦ－７细胞和ＥＲ阴性、表达突变型ｐ５３（ｍｔｐ５３）的ＭＤＡ－ＭＢ－２３１细胞以及ＥＲ转染阳性ＭＤＡ－ＭＢ－２３１细胞中ｐ５３、ｍｄｍ－２和ｐ２１ＷＡＦ１蛋白的表达水平，比较其与细胞生物学特性的关系。结果（１）ＭＣＦ－７细胞和ＭＤＡ－ＭＢ－２３１细胞ｐ５３蛋白的性质和分布明显不同，前者ｐ２１ＷＡＦ１和ｍｄｍ－２蛋白的表达水平明显高于后者（Ｐ＜０．０５），且前者的生物学特性较后者为好。（２）ＥＲ质粒转染ＭＤＡ－ＭＢ－２３１细胞后，其ｐ５３蛋白的表达水平降低（Ｐ＜０．０５），而ｍｄｍ－２蛋白的表达水平增加（Ｐ＜０．０５），生物学特性得以改善。结论乳腺癌细胞ＥＲ状态与ｐ５３和ｍｄｍ－２蛋白的表达水平以及生物学特性有关。     

小儿先天性心脏病肺静脉血流动力学改变

对２４例正常儿童及８８例先天性心脏病患儿的肺静脉血流动力学特征，进行了脉冲多普勒超声和彩色多普勒血流显像（ＣＤＦＩ）指标与手术指标的相关性研究。发现：房间隔缺损（ＡＳＤ）者肺静脉血流频谱（ＰＶＦ）呈单峰形特征，其峰值流速（Ｖｐ）与手术实测ＡＳＤ径高度正相关；其ＣＤＦＩ呈收缩期为著的五彩信号。室间隔缺损与动脉导管未闭者均呈高Ｄ峰的双峰形ＰＶＦ，Ｄ峰Ｖｐ与两病的手术实测径呈高度正相关；ＰＶＦ的ＣＤＦＩ呈舒张期为著五彩信号。表明小儿先天性心脏病时肺静脉血流动力学有不同的特征性改变。     

bcl-2、p53表达与乳腺癌预后的关系

目的：探讨ｂｃｌ２ 、ｐ５３ 表达与乳腺癌预后的关系。方法：应用免疫组化ＬＳＡＢ法检测６４ 例乳腺癌及３０ 例乳腺良性病变的表达。分析ｂｃｌ２、ｐ５３ 与乳腺癌组织学分级、腋淋巴结转移、复发和预后的关系。结果：ｂｃｌ２ 与ｐ５３ 表达之间差异有显著性,呈负相关（ Ｐ＜ ０－０５） 。ｂｃｌ２ 和ｐ５３ 表达与组织学分级有关（ Ｐ＜ ０－０５） ,ｂｃｌ２ 表达随分级增加阳性率降低,ｐ５３ 则相反。ｐ５３ 表达与腋淋巴结转移有关（ Ｐ＜ ０－０５） 。ｂｃｌ２ 表达与腋淋巴结转移无关（ Ｐ＞ ０－０５） 。ｐ５３ 表达复发组明显高于无复发组（ Ｐ＜０－０５）;ｂｃｌ２ 表达与有无复发无关（Ｐ＞０－０５）。ｐ５３ 表达阳性率≤５ 年生存组明显高于＞ ５ 年生存组,呈负相关（ Ｐ＜ ０－０５）;ｂｃｌ２ 表达与生存期无关（Ｐ＞ ０－０５） 。结论：ｂｃｌ２ 表达与预后无关,其阳性表达可反映肿瘤属分化较好或属早期阶段。ｐ５３ 可单独作为预后指标;ｐ５３ 表达与预后呈负相关。     

家兔急性心肌缺血后血清丙二醛变化及其与心肌梗塞面积的关系

本实验发现家兔急性心肌缺血后血中脂质过氧代物代谢产物丙二醛明显升高，且于２４．４８小时达到峰值，提示氧自由基在急性心肌缺血中起着重要作用。本研究还发现：４８小时血清丙醛水平与心肌梗塞面积呈正相关，为急性心肌梗塞诊断提供了辅助性指标。     

Differences in physical fitness among indoor and outdoor elite male soccer players

This study compared anthropometric (body height, body mass, percent body fat, fat-free body mass) and physical fitness characteristics (vertical jump height, power-load curve of the leg, 5 and 15 m sprint running time and blood lactate concentrations ([La]_b) at submaximal running velocities) among 15 elite male indoor soccer (IS) and 25 elite male outdoor soccer (OS) players. IS players had similar values in body height, body mass, fat-free body mass and endurance running than OS players. However, the IS group showed higher (P < 0.05–0.01) values in percent body fat (28%) and sprint running time (2%) but lower values in vertical jump (15%) and half-squat power (20%) than the OS group. Significant negative correlations (P < 0.05–0.01) were observed between maximal sprint running time, power production during half-squat actions, as well as [La]_b at submaximal running velocities. Percent body fat correlated positively with maximal sprint time and [La]_b, but correlated negatively with vertical jump height. The present results show that compared to elite OS players, elite IS players present clearly lower physical fitness (lower maximal leg extension power production) characteristics associated with higher values of percent body fat. This should give IS players a disadvantage during soccer game actions.     

微小RNA-181基因家族单核苷酸多态性和基因表达水平与缺血性脑卒中遗传易感性的关系

目的 探讨微小RNA（miR）-181基因家族rs16927589、rs77418916和rs8108402位点单核苷酸多态性（SNP）与缺血性脑卒中(IS)遗传易感性之间的关系;比较对照组与IS组中miR-181基因家族表达水平的差异,进一步探讨与基因表达水平之间的关系,为IS的防治工作提供帮助。 方法 使用SNaPshot技术对349例IS患者和372例对照组进行SNP基因分型检测,并用DNA测序法加以验证;使用日立7600生化仪检测对照组和IS组血脂水平;用ABI7500 Real-time PCR仪检测对照组和IS组外周血单核细胞miR-181基因家族的表达水平。 结果 关于rs8108402位点有CC、CT、TT 3种基因型,对照组和IS组基因型及等位基因频率对比发现,与CC基因型比较,CT基因型的携带人群患IS的风险性明显增高,TT基因型携带者患IS风险降低［CC vs CT:优势比（OR）=1.56,95%置信区间（CI）,1.11~2.18, P<0.05; CC vs TT: OR=0.25,95%CI, 0.10~0.62, P<0.01］,等位基因分析未发现相关性;rs16927589位点检测出TT、CT、CC 3种基因型,rs77418916位点有AA、AT、TT 3种基因型,对照组与IS组对比,未发现相关性。对rs8108402位点分层分析表明,携带CC基因型的IS患者其低密度脂蛋白（LDL-C）比携带CT基因型的IS患者高（P<0.05）,rs8108402位点多态性可能与IS的临床表现有相关性。IS组中,外周血单核细胞中miR-181a、miR-181b和miR-181c的表达明显高于对照组,差异有统计学意义（P<0.05）,而miR-181d的表达低于对照组,差异无计学意义（P>0.05）;对阳性位点rs8108402位点多态性与基因表达水平进一步分析发现,rs8108402位点多态性与基因表达水平无相关性。 结论 MiR-181c基因rs8108402位点CT和TT基因型可增加IS患病风险,CTC单倍型可增加IS患病风险。MiR-181c基因rs8108402位点多态性与LDL-C的高低有相关性,携带CC基因型的IS患者,其LDL-C水平较携带CT基因型患者高。MiR-181基因家族在正常对照组和IS组中的表达有明显差异,miR-181基因家族可能是IS的潜在的预测靶标和治疗靶基因。     

Development of a rapid PCR method using the insertion sequence IS1203 for genotyping Shiga toxin-producing Escherichia coli O157

We developed a rapid PCR method utilizing the diversity of the insertion site IS1203 for genotyping Shiga toxin-producing Escherichia coli (STEC) O157 (IS1203 PCR typing). DNA fragments digested by PvuII, which cut IS1203 at one site, were ligated with themselves and detected by PCR with outward-facing primer pairs for IS1203. To minimize nonspecific bands, nested PCR was also performed. Two fingerprinting patterns produced from the upstream or downstream regions of IS1203 were obtained within 1 or 2 days. By combining the two patterns, 79 STEC O157 isolates were classified into 39 types, which were then classified into 36 subtypes by pulsed-field gel electrophoresis (PFGE). The discriminatory power of IS1203 PCR typing (D = 0.974) is similar to that of PFGE (D = 0.981). This method can be used for rapid and simplified genotyping.     

Resetting the baroreflex during snoring: Role of resistive loading and intra-thoracic pressure

Baroreflex sensitivity (BRS) is reduced during snoring in humans and animal models. We utilised our rabbit model to examine the contribution of increased upper airway resistance to baroreflex resetting during snoring, by comparing BRS and baroreflex operating point (OP) values during IS to those obtained during tracheostomised breathing through an external resistive load (RL) titrated to match IS levels of peak inspiratory pleural pressure (Ppl). During both IS and RL, BRS decreased by 45% and 49%. There was a linear relationship between the change in Ppl and the decrease in BRS, which was similar for IS and RL. During both RL and IS, there was a shift in OP driven by ～16% increase in HR and no change in arterial pressure. Snoring related depression of BRS is likely mediated via a HR driven change in OP, which itself may be the outcome of negative intra-thoracic pressure mediated effects on right atrial wall stretch reflex control of heart rate.     

Rapid discrimination of Mycobacterium tuberculosis complex strains by ligation-mediated PCR fingerprint analysis.

A ligation-mediated PCR (LMPCR) method for the amplification of sequences flanking the IS6110 of the Mycobacterium tuberculosis complex has been developed. The method uses one primer specific for IS6110 and a second specific for a linker ligated to SalI-restricted genomic DNA. LMPCR is a rapid screening method, valuable for the fingerprinting of M. tuberculosis complex strains.     

Effects of melanotan II,a melanocortin agonist,on grooming and exploration in rats after repeated restraint/immobilization

2 mg/kg melanotan II (MTII, administered i.p.), a cyclic peptide analog of α-melanocyte stimulating hormone, at a single dose increased grooming in naive rats placed in an unfamiliar open-field device without changing locomotion or rearing. Male rats exposed to restraint/immobilization stress (IS) for 1 h on three consecutive days displayed increased grooming after the second stressor exposure, compared to pre-stress levels. MTII, administered to the rats after IS, enhanced the grooming response compared both to the pre- and post-stress values. The increase was greatest after the first dose and declined over the following two applications. As to the locomotion of rats in the entire experimental space, IS reduced the distance moved only after the first two stressor exposures; MTII did not influence these alterations. Locomotion in the central part of arena was not reduced by the stressor or by MTII, on the contrary, there was an increase in both groups after the third intervention. The only observed change in rearing was an increase in the MTII group after the third restraint exposure. Thus, MTII selectively increased grooming without markedly affecting the spatio-temporal structure of locomotor behavior in the open-field. The decline of MTII enhanced grooming over the three test days may be interpreted in terms of adaptation to the stressor and of the developing tolerance to the peptide.     

Different expression of positive and negative regulators of hepatocyte growth in growing and shrinking hepatic lobes after portal vein branch ligation in rats

Portal vein branch embolization is often performed before hepatectomy to prevent postoperative liver failure. It is, however, still not clear how the embolized lobe shrinks and the non-embolized lobe proliferates in counterbalance. We investigated the expression of positive and negative regulators of hepatocyte growth to clarify the mechanisms of liver growth and atrophy in a rat portal vein ligation (PVL) model compared with partial hepatectomy (PH). A significant increase in DNA synthesis within the non-ligated lobe reached a peak at 36 h, a delay of 12 h as compared with PH, while no increase occurred in the ligated lobe. Expression of hepatocyte growth factor mRNA remarkably increased in the non-ligated growing lobe between 6 and 24 h, but was only slightly elevated in the ligated shrinking lobe. Contrarily, negative regulators of hepatocyte proliferation, such as TGF-beta1 and IL-1beta, were strongly expressed in the ligated shrinking lobe. Thus, the changes of portal venous flow and/or pressure caused by PVL may contribute to induction of different kinds of growth factors between the ischemic and non-ischemic lobes; these factors possibly regulate liver regeneration and atrophy after PVL.     

Characterization of the 3rd International Standard for hepatitis B virus surface antigen (HBsAg)

BackgroundHBsAg is the most important marker for laboratory diagnosis of HBV infection. Validation and quality control of HBsAg tests requires International Standards (IS). Recently the 2nd IS was replaced by the 3rd IS. Both IS are made from plasma-derived hepatitis B vaccines, but production and geographical origin are different.ObjectiveCharacterization of the HBsAg in the source material (SM) for the 3rd IS and comparison with the 2nd IS and native HBsAg.Study designThe SM was analyzed using solid-phase immunoassays, quantitative immune electrophoresis, ultracentrifugation, immunoblotting and HBV DNA sequencing.ResultsThe plasma-derived HBsAg of the SM originated from at least two different HBV strains, both of subgenotype (sgt) B4, typical for Vietnam. The HBsAg subtype was heterogeneous with ayw1 and adw2. The HBsAg concentration was 23,700 IU/ml as determined by solid-phase immunoassay; immune electrophoresis calibrated with sgt B2 revealed a concentration of 24,500 IU/ml while calibration with sgt D1 provided lower values. Proteins in the SM are heterogeneous in size containing only traces of preS. The protein subunits are partially cross-linked.ConclusionsThe antigenicity of the 3rd IS is suitable for HBsAg calibration in laboratory tests. In contrast to the 2nd IS, the 3rd IS is representative for a highly endemic region. Similar to the 2nd IS and different from native HBsAg, preS domains are depleted, protein subunits are partially cross-linked and the HBsAg particles are partially aggregated in the 3rd IS. The HBV subgenotype differences between the two IS may lead to variations in different quantitative assays.