Rapid identification and susceptibility testing using the VITEK(R) 2 system using culture fluids from positive BacT/ALERT(R) blood cultures

BACKGROUND AND PURPOSE: In order to reduce the turnaround time for laboratory diagnosis of bacteremia, the efficacy of identification and antimicrobial susceptibility testing using samples taken directly from positive BacT/ALERT(R) standard aerobic and standard anaerobic blood culture bottles was evaluated. METHODS: 160 positive blood culture bottles were examined and incubated at 35 degrees C in 5% carbon dioxide for 4-24 h, and an aliquot of the culture fluid was Gram stained. Samples containing Gram-negative bacilli were inoculated on VITEK(R) 2 ID-GNB (identification-Gram-negative bacilli) and AST (antimicrobial susceptibility testing)-GN04 cards, and those containing Gram-positive cocci were inoculated on ID-GPC (identification-Gram-positive cocci) and AST-P526 cards. The same samples were also examined by the standard method, involving subculture from positive BacT/ALERT standard blood culture bottles. RESULTS: Eighty seven of 97 Gram-negative bacilli (89.7%) and 21 of 63 Gram-positive cocci (33.3%) were correctly identified to the species level. For antimicrobial susceptibility testing, the direct method had an overall error rate of 5.4% for Gram-negative bacilli, with 0.9% very major, 0.9% major, and 3.6% minor discrepancies compared to the standard method. The overall error rate in antimicrobial susceptibility testing for the 13 Staphylococcus spp. was 10.3%, with 6.0% very major, 2.6% major, and 1.7% minor discrepancies. CONCLUSION: These data suggest that VITEK 2 cards inoculated with samples taken directly from positive Bact/ALERT blood culture bottles would provide acceptable identification and antimicrobial susceptibility testing results for Gram-negative bacilli, but not for Gram-positive cocci. Compared to the standard method, the direct method would reduce turnaround time by at least 24 h.     

Intratracheal administration of liposomal clodronate accelerates alveolar macrophage reconstitution following fetal liver transplantation

To facilitate study of alveolar macrophages in vivo, we developed a method to rapidly and efficiently replace resident alveolar macrophages with macrophages of a different (donor) genotype. Chimeric mice were generated by lethal irradiation followed by fetal liver transplantation (FLT) using green fluorescent protein (GFP) transgenic reporter mice as donors. Kinetics of peripheral blood monocyte (PBM) and alveolar macrophage reconstitution was determined 4 and 10 weeks post-FLT by quantifying the percentage of GFP+ cells. To enhance the recruitment of donor monocytes into the lung after FLT, mice were treated with intratracheal administration of liposomal clodronate to deplete host alveolar macrophages at 6 weeks post-FLT. PBM reconstitution occurred by 4 weeks after FLT (85.7+/-1.6% of CD11b+/Gr-1+ monocytes were GFP+), and minimal alveolar macrophage repopulation was observed (9.5% GFP+). By 10 weeks following FLT, 48% of alveolar macrophages were GFP+ by immunostaining of macrophages on lung tissue sections, and 55.1 +/- 1.6% of lung lavage macrophages were GFP+ by fluorescein-activated cell sorter analysis. Clodronate treatment resulted in a significant increase in GFP+ alveolar macrophages 10 weeks after FLT. By immunostaining, 90% of macrophages were GFP+ on lung tissue sections and 87.5 +/- 1.1% GFP+ in lung lavage (compared with GFP-transgenic controls). The ability of newly recruited alveolar macrophages to clear Pseudomonas aeruginosa and activate nuclear factor-kappaB in response to Eschericia coli lipopolysaccharide demonstrated normal macrophage function. Optimizing this methodology provides an important tool for the study of specific genes and their contribution to alveolar macrophage function in vivo.     

神经系统疾病少年儿童患者的脆性X综合征检查与临床诊治研究

目的研究神经疾病少年儿童患者脆性X综合征（fragjle X syndrome,FraX）的细胞遗传学检测和临床诊治特征。方法应用低叶酸培养基，对患神经疾病的18岁以下146例儿童行X染色体脆性部位的检测，对检出的Frax患儿给以叶酸为主辅以营养神经和神经康复的治疗。结果146例就诊患儿检出FraX13例，检出率为8．90％。以补充叶酸为主的综合治疗取得一定疗效，治疗效果与患儿年龄和原发神经疾病有关。结论FraX值得临床重视，早检出、早治疗收效较好。     

SARS冠状病毒棘突蛋白的S1蛋白诱导小鼠产生中和抗体的研究

目的 研究SARS冠状病毒棘突蛋白受体结合部位S1的免疫原性，为SARS的实验诊断和新型疫苗的研究提供依据。方法 用克隆有哺乳动物细胞密码子优化的SARS-CoV S1基因的质粒pcDNA3．1／S1或P-S1Ig转染293T细胞，用细胞的上清液纯化S1蛋白。以pcDNA3．1／S1质粒对BALB／c小鼠进行2次基因免疫，以纯化的S1蛋白进行加强免疫。用ELISA法检测小鼠抗SARS-CoV的特异性IgG抗体，并在Vero E6细胞上做体外中和实验，检测中和抗体。结果 S1蛋白诱导小鼠产生抗SARS-CoV的特异性抗体；1：1499．68稀释的S1蛋白免疫的小鼠血清可保护50％的细胞对1000TCID50的病毒攻击，而阴性对照血清不能保护细胞对病毒的感染。结论 SAPS冠状病毒棘突蛋白受体结合部位S1能有效诱导机体产生具有高效保护作用的中和抗体免疫反应，可望发展成为理想的SARS棘突蛋白亚单位疫苗。     

氟烷和七氟醚对缺血心肌功能和代谢及Ca2+-ATP酶活性的影响

目的: 研究氟烷、七氟醚(1.5MAC)对缺血心肌的影响。方法: 应用离体大鼠心脏Langendorff逆行灌注模型研究氟烷、七氟醚对心肌缺血前心率(HR)、左室舒张末期压力(LVEDP)、左室发展压(LVDP)、左室压力升高速率(+dp/dt)、左室压力下降速率(-dp/dt)和冠脉流量(CF)的影响,测定缺血前、缺血10min、缺血25min3个不同时间的心肌ATP含量、Ca²⁺-ATP酶活性,同时记录缺血间期左室内压的变化情况。结果: 七氟醚显著增加正常离体心脏的CF,氟烷、七氟醚均不同程度地抑制心肌收缩功能和Ca²⁺-ATP酶活性,能够增加正常心肌的能量贮备。缺血10min时,二药能够减缓心肌ATP含量及Ca²⁺-ATP酶活性的下降,氟烷的作用比较明显。缺血间期,氟烷明显推迟缺血性挛缩的起始时间,降低挛缩幅度。结论: 氟烷的抗缺血损伤作用优于七氟醚,延缓缺血期心肌ATP含量及Ca²⁺-ATP酶活性的下降可能是氟烷抗缺血损伤作用的重要机制之一。     

"Waste" follicular aspirate from fertility treatment--a potential source of human germline stem cells?

Fertility clinics worldwide routinely produce a large volume of 'waste' follicular aspirate, which is potentially an abundant source of immature ovarian follicles. Current attempts to cultivate these further in vitro to yield viable mature oocytes for fertility treatment have not yet achieved much success. Instead, recent lines of evidence have emerged that are suggestive of a potential stem cell niche within such immature ovarian follicles. The recent discovery of follicular renewal and putative germ-line stem cells within the postnatal mammalian ovary shook the foundations of reproductive biology by challenging the established dogma that mammalian females lose the capacity for germ cell renewal during fetal life, such that a fixed reserve of germ cells (oocytes) enclosed within follicles is endowed at birth. More intriguingly, another recent study in the Drosophila model provided compelling evidence that somatic progenies (nurse cells) of germ-line stem cells had the ability to revert back to the stem-cell-like state. This introduces the exciting possibility that within the mammalian ovarian follicle, similar somatic progenies of germ-line stem cells may also possess a greater intrinsic ability to revert back into functional stem cells. If this is the case, then a favored candidate would be the cumulus/granulosa of immature ovarian follicles, since such cells are true homologues of nurse cells found within the Drosophila ovary. The successful elucidation of a human germ-line stem cell niche within immature ovarian follicles is likely to have huge ramifications in stem cell biology and regenerative medicine.     

肠系膜上静脉外科干及下腔静脉下段的应用解剖观察

本文解剖了30具成人尸体,对肠系膜上静脉外科干、下腔静脉下段的位置、长度、外径、两者的相互关系以及前者与肠系膜上动脉分支后者与腹主动脉分支的关系等进行了观察测量。认为国人的外科干至少80％能满足手术中行肠腔静脉吻合术应具备的四点要求。     

Synthesis and characterization of amphiphilic and microphase-separated graft copolymers, 1. Synthesis and bulk characterization of polystyrene-graft-ω-stearyl-poly(ethylene oxide)

Using the rigid and hydrophobic polystyrene (PS) chain as backbone, onto which flexible and hydrophilic stearyl-poly(ethylene oxide) (SPEO) chains are grafted, a new kind of amphiphilic, microphase-separated graft copolymer was synthesized using the macromonomer technique. Stearyl-poly(ethylene oxide) macromonomers with acryloyl end-group (SPEO-A) were prepared through an end-group exchange reaction of α-stearyl-ω-hydroxypoly(ethylene oxide) (SPEO-OH) and acryloyl chloride in the presence of triethylamine. The radical copolymerization of styrene with SPEO-A was carried out under various experimental conditions. Following a careful examination of their purity, the structure of the prepared copolymers was characterized by means of IR, ¹H NMR and GPC analyses. A new feasible method using first derivative UV spectrometry was developed for quantitative determination of the bulk composition of the graft copolymers. Copolymers with a wide range of bulk composition and satisfactory grafting degree were obtained.     

中国汉族人与美国高加索白人膝关节几何形态的比较

背景:有研究显示中国人膝关节胫骨近端前后径和横径均小于美国高加索人,所以在膝关节形态学方面存在一定差异,因此常发生进口假体系统应用于国人后出现匹配度不高的现象。目的:测量中国人和美国人股骨远端、胫骨近端的几何形态学参数,比较其差异。方法:选择北京清华长庚医院骨科拟行前交叉韧带重建的中国汉族患者50例,以及美国匹兹堡大学运动医学中心拟行前交叉韧带重建的高加索白人后裔患者40例,对90例受试者膝关节进行CT扫描,使用AW Volume Share 5软件进行三维重建,利用Geomagic软件对胫骨近端和股骨远端的各项参数进行测量。结果与结论:①股骨近端测量参数中,中国人的外侧胫骨平台内外径小于美国人(P=0.027),外侧胫骨平台后倾角大于美国人(P<0.05);两组间胫骨平台内外径、内侧胫骨平台外径、内侧胫骨平台前后径、外侧胫骨平台前后径与内侧胫骨平台后倾角比较差异均无显著性意义(P>0.05);②股骨远端测量参数中,中国人的股骨远端横径、外侧髁内外径、内侧髁前后径、外侧髁前后径及髁间窝高度均小于美国人(P<0.05),股骨外翻角大于美国人(P<0.05);两组间内侧髁内外径、髁间窝宽度及滑车沟宽度比较差异无显著性意义(P>0.05);③结果表明,中国人与美国人在膝关节形态学的多个参数存在差异,有必要针对中国人设计更为个体化的膝关节假体。