20株阴沟肠杆菌耐药性及氨基糖苷类修饰酶基因分析

目的明确临床分离的20株阴沟肠杆菌耐药性及氨基糖苷类修饰酶基因存在状况。方法采用ATB药敏试验板微量肉汤法测定临床分离的20株阴沟肠杆菌对20种抗菌药物的敏感性，采用聚合酶链反应及序列分析的方法分析氨基糖苷类修饰酶基因型。结果该20株菌呈现多重耐药，对亚胺培南和美罗培南均敏感，对阿莫西林、阿莫西林／克拉维酸、头孢噻吩和头孢西丁完全耐药，对头孢吡肟及4种氨基糖苷类抗生素阿米卡星、庆大霉素、妥布霉素和奈替米星的耐药率分别为25．0％、60．0％、85．0％、90．0％和90．0％，其余9种的耐药率在80．0％～95．0％之间。19株(95．0％)检出氨基糖苷类修饰酶基因；aac(6′)-Ⅰ、aac(3)-Ⅱ、ant(3″)-Ⅰ、ant(2″)-Ⅰ和aph(3′)-Ⅵ基因的阳性率分别为80．0％、50．0％、40．0％、5．0％和5．0％；而aog2(3)-Ⅰ、aac(3)-Ⅲ、aac(3)-Ⅳ和aac(6′)-Ⅱ基因均阴性。结论临床分离的阴沟肠杆菌多重耐药严重，氨基糖苷类修饰酶基因携带率很高。     

CD3+木瓜提取物对ApoE基因缺陷小鼠血脂及炎性相关因素的影响

目的观察木瓜提取物对ApoE基因缺陷小鼠血脂水平、主动脉窦斑块中巨噬细胞活化及肿瘤坏死因子(TNF-α)水平的影响。方法30只雄性ApoE基因缺陷小鼠随机分为3组:对照组、低剂量组和高剂量组,低剂量组和高剂量组分别在基础饲料上添加5%和10%的木瓜提取物喷雾干燥颗粒,饲养16周后经小鼠眼眶静脉取血后处死动物,酶法测定血脂水平、采用免疫组化分析主动脉窦斑块中TNF-α的表达水平和采用免疫荧光反映斑块CD68阳性表达水平与巨噬细胞阳性表达率。结果低剂量组和高剂量组的血清TC和LDL-C显著低于对照组(P<0.05),木瓜提取物能显著下调斑块中TNF-α的表达水平以及减少巨噬细胞的阳性率。结论木瓜提取物降低了血清脂蛋白胆固醇水平(TC和LDL-C),减少了炎性因子TNF-α表达以及减弱巨噬细胞参与程度,预示其可能具有减缓动脉斑块发生发展的抗动脉粥样硬化作用。     

含氮二膦酸盐/IL-2诱导肿瘤患者外周血γδT细胞增殖的作用

目的探讨药物（含氮二膦酸盐联合IL-2，N-BP／IL-2）诱导实体肿瘤患者外周血γδT细胞增殖作用。方法采用流式细胞仪测定103例肿瘤患者和43例健康人外周血总T细胞绝对值、御细胞绝对值和相对值，测定患者外周血经药物体外刺激后御细胞增殖指数和用药前后患者外周血总T细胞、γδT细胞绝对值和相对值。结果肿瘤患者和健康人外周血总T细胞绝对值、御细胞绝对值和御细胞相对值的中位数，分别为1306细胞/μl和1522细胞/μlP=0．003），64细胞/μl和162细胞/μl（P〈0．001），5％和11％（P〈0．001）。24例经药物体外扩增试验阳性的肿瘤患者治疗前后体内外周血总T细胞绝对值、γδT细胞绝对值和御细胞相对值的中位数，分别为1283细胞/μl和1552细胞/μl（P=0．001），54细胞/μl和107细胞/μl（P〈0．001），6％和8％（P=0．008）。全部肿瘤患者中外周血γδT细胞对药物体外诱导增殖反应阳性患者的百分率为68．9％。结论肿瘤患者外周血γδT细胞计数显著低于健康人。不同肿瘤患者外周血γδT细胞对药物刺激增殖的反应性不同。N-PB／IL-2在体内有显著增加御细胞数量的药理作用。体外增殖试验可作为N-PB／IL-2治疗患者的参考。     

Andrology: Acrosome reaction inducibility predicts fertilization success at in-vitro fertilization

We prospectively studied the ability of acrosome reaction (AR)inducibility to predict fertilization success in a group of232 infertile patients presenting sequentially for in-vitrofertilization (IVF). The median percentage of eggs fertilizedfor the overall patient population was 25% (interquartile range5–58%), with one to 29 oocytes available for insemination(median, five oocytes). The median percentage of eggs fertilizedat IVF increased as the percentage of spermatozoa able to undergoAR became greater: spermatozoa with a failed AR (5%) fertilizedonly 12% of eggs, while spermatozoa with AR values>9% fertilized50% of eggs. The assay had a specificity of 0.75, a sensitivityof 0.55 and an odds ratio of 2.9; thus, AR-positive patientsare 2.9 times more likely to achieve fertilization than patientswith a failed AR. Receiver operator characteristic (ROC) curveswere constructed for AR, sperm concentration and percentageof normal forms in semen. All three parameters proved to bepotentially useful in predicting the occurrence of fertilization,although AR and morphology appeared to be better than spermconcentration by ROC analysis. Patients were divided into fourclearly defined subgroups according to their traditional semencharacteristics, including morphology. The median percentageof eggs fertilized decreased as traditional semen characteristicsdeteriorated, from a median of 46% for patients with excellentsperm concentration, motility and morphology, to a median of29% for patients with suboptimal semen quality and a medianof 0% for patients with severely impaired semen. Within eachpatient subgroup, the median percentage of eggs fertilized was3-to 4-fold higher for individuals with a positive AR than forthose with a failed AR, indicating that AR has a greater effecton fertilization rate than traditional semen parameters includingmorphology. We now recognize that some men with good semen characteristicshave an unexpectedly poor AR and a markedly reduced fertilizationrate, while other men with poor traditional semen characteristicsunexpectedly retain AR and perform relatively well at IVF. Bycontrast to AR, morphology seemed to have little effect on fertilizationsuccess (two-way analysis of variance not significant). Thewife's age and oocyte quality were evenly distributed amongthe different patient subgroups, indicating that differencesin fertilization rate could not be attributed to either parameter.Our data indicate that AR has a much higher predictive valuefor IVF success than traditional semen parameters includingmorphology. We propose that AR assessment is a clinically usefuldiagnostic tool in determining a patient's likelihood of achievingfertilization at IVF.     

二十碳五烯酸联合视黄酸对HL-60细胞凋亡相关基因表达的影响

目的 研究二十碳五烯酸(EPA)是否协同视黄酸(RA)影响HL-60细胞的凋亡及相应分子机制。方法 流式细胞仪(FCM)检测细胞周期相分布以测定细胞增殖和凋亡功能；Western blot法分析bcl-2和caspase—3表达。结果 与单独应用相比，EPA和RA联合应用可增强HL-60细胞的调亡效应，以及下调节bcl—2和增强caspase-3基因表达。结论 EPA和RA联合应用对HL-60细胞凋亡的增强效应，可能与它们增强caspase-3和降低bcl—2基因表达相关。     

细胞内外膜跨膜电位频率响应模型及滤波器特性的研究

提出球形单细胞内外膜跨膜电位频率响应模型和仿真计算方法。通过对频率响应的仿真分析，发现内外膜跨膜电位分别表现出一阶带通和低通滤波器特性，并在内膜频率响应中心频率到上限截止频率的范围内，内膜跨膜电位大于外膜同时能保持在较高水平，这将有利于诱导细胞内电处理效应。计算结果与动物实验吻合得很好，为纳秒脉冲电场治疗肿瘤的窗口参数合理选择提供了理论依据。     

纳秒级陡脉冲电场诱导癌细胞凋亡的实验及作用机理研究

观察纳秒级陡脉冲电场诱导人浆液囊腺性卵巢癌SKOV3癌细胞凋亡效应,及其对细胞内钙离子浓度即[Ca2＋]i的影响并探讨其作用机理。采用场强为10kV/cm、脉宽为100ns、重复频率为1Hz的纳秒级陡脉冲电场作用于SKOV3癌细胞悬液5min。4h后采用Annexin V-FITC/PI双标记并结合流式细胞仪分析SKOV3癌细胞的凋亡率,采用扫描和透射电子显微镜观察癌细胞凋亡的超微结构变化;以Fluo-3/AM为细胞内钙离子的荧光指示剂,用激光扫描共聚焦显微镜检测纳秒级陡脉冲电场对SKOV3癌细胞[Ca2＋]i的影响,及[Ca2＋]i变化时的Ca2＋来源。结果显示,流式细胞术检测到癌细胞的早期凋亡率为（22.21&#177;2.71）%,明显高于对照组的（3.04&#177;0.44）%（P〈0.01）,扫描和透射电子显微镜均观察到典型的凋亡细胞形态,证实纳秒级陡脉冲电场能够有效诱导SKOV3癌细胞发生凋亡;同时纳秒级陡脉冲电场可明显升高细胞[Ca2＋]i（P〈0.01）,而与细胞外钙离子浓度无关（P〉0.05）。由于纳秒级陡脉冲电场的等值频率很高,容易穿透细胞膜,其诱导凋亡的机制之一可能是通过激活细胞内钙库（内质网、线粒体）,引起细胞内钙离子升高,从而介导凋亡信号通路。     

Surface receptors on neutrophils and monocytes from immunodeficient and normal horses.

Surface receptors on peripheral blood neutrophils and monocytes from normal and immunodeficient horses have been studied. Sheep erythrocytes (SRBC) coated with IgG, IgM, and complement but not IgG(T), readily bound to normal equine monocytes and neutrophils. More than 4000 molecules of IgG were required to sensitize each SRBC for adherence to monocytes, and more than 12,000 molecules were required for adherence to neutrophils. Young horses with a severe combined immunodeficiency had an almost total absence of lymphocytes, but normal numbers of monocytes and neutrophils. The number of receptors for immunoglobulin, complement, and phytolectin on monocytes and neutrophils from immunodeficient animals were similar to those on the cells of normal horses. Although the precursor cells of lymphocytes of horses with combined immunodeficiency appear to be defective, no defect in the other cellular products of the bone marrow were apparent.