高滴度呼肠孤病毒3型的制备及滴度检测

 目的   制备高滴度呼肠孤病毒3型（reovirus 3,Reo-3）病毒液,并检测Reo-3滴度。 方法   以幼仓鼠肾细胞（BHK-21）为病毒培养基质和滴度测定细胞。将Reo-3按10⁰至10^-5感染复数（MOI）接种BHK-21,再分别用细胞病变法和蚀斑形成法检测病毒,用Karber法计算病毒滴度。 结果   BHK-21感染Reo-3后能产生明显病变。以10^-4 MOI的Reo-3接种后48 h,收获液中病毒滴度最高,细胞病变法检测为9.625 lgTCID50/ml,蚀斑形成法检测为8.671 lgPFU/ml。 结论   制备了高滴度Reo-3,为开展该病毒去除灭活研究奠定了基础。     

基于呼肠孤病毒的治疗有望用于晚期实体瘤

据国际癌症生物药治疗学会年会上提出的一项研究，OncolytcsBiotech公司的专利的人呼肠孤病毒制剂Reolysin（I）与化疗药物合用有潜力用于治疗晚期实体瘤。     

生物素化单克隆抗体的亲和素-生物素放大荧光ELISA在病毒的定性及定量分析中的应用

ELISA虽是一种快速有效的检测各种病毒的诊断工具,然而在某些情况下,如直接测定体液中低浓度的病毒抗原或未经预先浓缩的病毒,则不够灵敏。本文作者建立了一种用维生素生物素和糖蛋白亲和素之间的高亲和力多价结合反应放大的荧光ELISA(FELISA)方法,这种放大的FELISA的特异性用同种或异种新城疫病毒(NDV)株和其他血清型相关或不相关的病毒来评估,发现试     

血液病毒核酸检测技术在我国的应用现状

核酸检测技术（NAT）是直接检测病原体核酸的一系列技术的总称，是一种灵敏度、特异性高和检测通量大的血液病毒检测方法，可明显缩短血液病毒检测窗口期并能检测出因病毒变异而漏检的血液，显著提高输血安全。本文对血液病毒核酸检测技术在我国的应用现状进行综述，以期为我国血站开展血液病毒核酸检测在方法选择、检测系统选择、实验室的建设和管理、检测模式等方面提供参考。     

电镜技术在植物病毒研究中的应用

本文简要介绍了目前常用于植物病毒研究的5种电镜制样方法及其应用。     

TT病毒结构蛋白抗原在大肠杆菌中的表达及应用

目的　用原核系统表达TT病毒 (TTV)的重组蛋白抗原 ,建立诊断 (TTV)感染的特异性方法。方法　采用PCR方法扩增TTVORF2 基因 ,将之插入到测序质粒载体中进行测序 ,验证序列正确后将之嵌入到原核表达载体pQE30中 ,并转化大肠杆菌M15菌株 ,进行诱导表达。用SDS -PAGE分析表达产物。表达产物经Ni-NTA柱层析纯化后 ,得到纯度为 90 %的ORF蛋白抗原。用TTV感染者的阳性血清对该蛋白进行了West ern -Blot分析 ,用间接ELISA方法检测血清抗 - (TTV)IgG抗体。结果　经IPTG诱导后 ,筛选出 2株高表达株。结论　该蛋白具有良好的抗原活性 ,可以有效地用于TTV的感染检测中     

前S1抗原检测在乙型肝炎诊断中的应用

目的 通过对乙型肝炎病毒(HBV)前S1抗原与HBV血清标志物检测的对比分析,对HBV前S1抗原的临床意义进行探讨.方法 用酶联免疫吸附实验(ELISA)对HBV血清标志物和HBV前S1抗原进行检测.结果 对HBV血清标志物不同组合模式进行分析发现:HBV血清标志物传染性强(HBeAg阳性)的模式中,HBV前S1抗原的检出率为94.0%;HBV血清标志物传染性弱(HBeAg阴性)的模式中,HBV前S1抗原的检出率为15.6%;乙肝表面抗原(HBsAg)、乙肝e抗原(HBeAg)均为阴性的模式中,HBV前S1抗原均是阴性.结论 HBV前S1抗原与HBeAg在很大程度上具有一致性,其阳性可作为HBV复制的重要依据和HBV存在的指标.     

直接免疫荧光法病毒抗原测定在呼吸道感染性疾病中的诊断应用

目的：研究直接免疫荧光法病毒抗原测定在呼吸道感染性疾病中的诊断应用。方法呼吸道感染的患者690例,采取患者鼻咽深部的分泌物,应用直接免疫荧光法测定呼吸道腺病毒（ADV）、合胞病毒（RSV）、流感病毒A型（InfA）、流感病毒B型（InfB）以及副流感病毒（PIV）等7种病毒的抗原。结果呼吸道病毒检测阳性者240例,占34.8%,其中呼吸道腺病毒（ADV）12例,占5.0%,合胞病毒（RSV）169例,占70.4%,流感病毒A型（InfA）18,占7.5%,流感病毒B型（InfB）2例,占0.8%以及副流感病毒（PIV1）39例,占16.3%。结论直接免疫荧光法病毒抗原测定可以用于呼吸道病毒感染诊断。     

四参数模型和平行线分析法在病毒疫苗抗原生物检定统计中的应用

目的 比较四参数模型与平行线分析法在Sabin株脊髓灰质炎灭活疫苗(Sabin inactivated poliovirus vaccine,sIPV)D抗原和肠道病毒71型(enterovirus 71,EV71)抗原生物检定统计中的应用.方法 对sIPV-D抗原和EV71抗原检测数据分别进行四参数模型和平行线分析法的方法验证,并将两种方法应用于不同含量样品检测结果的分析计算.结果 sIPV-D抗原检测数据四参数曲线拟合度好(决定系数为0.998,残差值≤0.059),平行性分析可靠(线性回归:参比品F=727.49,P＜0.01;样品F=1 169.44,P＜0.01.平行性方差分析F=0.24,P＞0.05).EV71抗原四参数曲线拟合度好(决定系数为1.000,残差值≤0.025),平行性分析可靠(线性回归:参比品F=126.63,P＜0.01;样品F=192.74,P＜0.01.平行性方差分析F=4.00,P＞0.05).对于sIPV和EV71不同抗原含量的生物检定统计,两种方法计算结果的差异无统计学意义(t=0.248～1.023,P值均＞0.05).结论 四参数模型和平行线分析法均能较好地应用于sIPV-D抗原和EV71抗原的ELISA检测体系.     

符合三酶标准的胰酶生产乙醇法工艺研究

作者采用选择适当、适量的激活剂,改变提取激活的溶剂、时间、温度等措施,对国内原用猪胰酶生产工艺进行改革,使产品质量达到中国药典(85)版规定的三酶标准,生产周期缩短,收率也有所提高。     

Human ABC transporters ABCG2 (BCRP) and ABCG4

1.?The human ABC transporter ABCG2 is regarded as a member of the phase III system for xenobiotic metabolism, and it has been suggested that this efflux pump is responsible for protecting the body from toxic xenobiotics and for removing metabolites.

2.?This review paper will address the new aspects of ABCG2 in terms of post-translational modifications (i.e., disulfide bond formation, ubiquitination, and endoplasmic reticulum-associated degradation) of ABCG2 protein, high-speed screening, and quantitative structure–activity relationship (QSAR) analysis to evaluate ABCG2–drug interactions, and genetic polymorphisms potentially associated with photosensitivity.

3.?In addition, new aspects of human ABCG4 and mouse Abcg4 are presented with respect to their molecular properties and potential physiological roles. Considering a high sequence similarity between ABCG1 and ABCG4, both Abcg4 and ABCG4 may be involved in the transport of cholesterol from neurons and astrocytes. Furthermore, high expression of the mouse Abcg4 protein in the testis implicates its involvement in transport of certain sex hormones.     

THE USE OF KNOCKOUT MICE TO UNDERSTAND THE ROLE OF CYTOKINES IN FEVER

1. In most instances, data obtained using knockout mice to dissect the role of cytokines in fever are similar to data obtained by other, more traditional experimental techniques. 2. Interleukin (IL)-lβ appears to be critically involved in fever caused by some routes of infectiodinflammation (e.g. localized inflammation with turpentine). This cytokine has only a small role in fevers caused by i.p. injection of lipopolysaccharide (LPS). These IL-lβ-induced fevers in knockout mice appear to be via the induction of IL-6, similar to LPS-induced fevers in rats. Interleukin-6 also appears to be critically involved in turpentine-induced fever. 3. The precise role of tumour necrosis factor (TNF) in fever is controversial. Data obtained from knockout mice lacking both TNF receptors do not support a pyrogenic role for TNF in fever either to i.p. injection of LPS, s.c. injection of turpentine or following caecal ligation and puncture. 4. The roles of these cytokines in fevers induced by injection of LPS, IL-lβ, turpentine and caecal ligation and puncture are summarized. The data show the complexity of the febrile response. Depending on the types of inflammatoryhnfectious stimuli, different cytokines play important roles. Because other cytokines are thought to be involved in fever (e.g. macrophage inflammatory protein, interferons), considerable work is still needed to dissect the precise roles of cytokines in fever.     

THE USE OF A RAT ISOLATED ILEAL PREPARATION TO INVESTIGATE THE RELEASE OF NEUROTENSIN

1. An isolated and perfused preparation of rat ileum was used to investigate the effects of cholinergic, adrenergic and bombesin stimuli on neurotensin release into the vascular compartment. 2. A vigorous release of neurotensin like immunoreactivity (NTLI) to 200% above basal values in response to intraluminal infusions of emulsified soybean oil (Intralipid) demonstrated the physiological responsiveness of the preparation. 3. Carbachol significantly stimulated the release of both NTLI and bombesin like immunoreactivity (BLI), with maximal responses at 5 times 10^-9 mol/l carbachol of 100% and 400% above basal values for NTLI and BLI, respectively. 4. Noradrenaline at 10^-6 and 10^-4 mol/l caused no significant release of NTLI but markedly inhibited spontaneous BLI release. 5. Synthetic amphibian bombesin caused a marked release of NTLI to 81% and 100% above basal at infusion concentrations of 5 times 10^-11 and 5 times 10^-10 mol/l respectively. 6. These results suggest that there is either a direct effect of cholinergic agents on the N cells bringing about NTLI release, or an indirect effect via the release of bombesin like peptides from intrinsic gut neurones. This preparation provides a useful model to study the complex neural, paracrine and endocrine interactions in the gastrointestinal tract.     

血浆中甲基还原青蒿素(蒿甲醚)的薄层扫描定量法

甲基还原青蒿素(MDHA)是青蒿素的衍生物,经药理及临床试验证实其抗疟疗效优于青蒿素,为了开展体内过程及药代动力学研究,使给药方案更加科学合理,急需建立生物样品的测定方法。本文报道血浆中MDHA的薄层扫描定量法。本法特异性较高,检出下限为0.1μg,CV低于13.0%,血浆回收率达83%以上。     

应用导数光谱法考察苯呋洛尔溶液体外经皮渗透性

本文应用二阶导数紫外分光光度法考察了苯呋洛尔溶液体外经皮渗透性。考察了促进剂月桂氮(?)酮-丙二醉及PEG-400溶剂对其渗透性的影响。应用二阶导数紫外分光光度法消除了兔皮溶出物对一般紫外测定方法的干扰。结果表明:苯呋洛尔溶液具有体外经皮渗透性;月桂氮(?)酮-丙二醇系统及PEG-400均能增强苯呋洛尔溶液的体外经皮渗透性。     

SYNTHESIS AND BIOLOGICAL EFFECT OF THE C-TERMINAL PENTAPEPTIDE AMIDE OF AVIAN PANCREATIC HORMONE III (APP)

Model building studies and analogies drawn from peptides of similar biological activity have indicated that the C-terminus of avian pancreatic hormone III may possess significant biological information. To test this hypothesis, the C-terminal pentapeptide amide sequence has been synthesized by the Merrifield method. The synthesis, purification and characterization of this compound are reported here in detail. In vivo studies indicate that this synthetic segment possesses none of the secretogogic activity of the parent hormone; rather, it reduces “gastric” secretion levels even in the presence of the intact hormone.     

USE OF THE CHROMOGENIC p-(p-(DIMETHYLAMINO) PHENYLAZO) BENZYL (AZ) ESTER IN THE SYNTHESIS OF LEU-ENKEPHALIN

Solution synthesis of Leu-enkephalin has been achieved using the coloured carboxyl protecting p-(p-(dimethylaminojphenylazo)benzyl moiety throughout the synthesis. The intense colour of this protecting group allowed facile purification of protected intermediates on silica gel or ion exchange columns and visualisation on thin-layer chromatography. The chromogenic p-(p-fdimethyl-amino)phenylazo)benzyl ester group readily withstands successive treatments with 25% trifluoroacetic acid in dichloromethane yet is cleanly removed by catalytic hydrogenation.     

CHARACTERIZATION OF AN APPROACH TO DEVELOPMENTAL IMMUNOTOXICOLOGY ASSESSMENT IN THE RAT USING SRBC AS THE ANTIGEN

Although developmental immunotoxicology is an area of emerging importance, few data exist that compare profiles of activity in neonatal and adult rodents. One of the factors contributing to the paucity of comparative results is that it is unclear whether immunotoxicological test procedures optimized to evaluate the immune system of adults can be integrated into studies of younger animals. Therefore,the following objectives were addressed in this study for both male and female Crl:CD(SD)BR rat pups and weanlings: (1) to quantify baseline values for absolute and relative splenic lymphocyte populations using flow cytometry in nonimmunized animals; (2) to optimize and establish baseline values for the primary humoral immune response to sheep red blood cells (SRBC) using either an enzyme-linked immunosorbant assay (ELISA) or the plaque-forming cell (PFC) assay; and (3) to determine the impact of SRBC immunization on the histology of the spleen. Responses for each age group were compared both withinandbetween litters. Spleencell counts and absolute andrelative (to body weight) spleen and thymus weights were also obtained. Analysis by flow cytometry indicated the following overall mean absolute and relative (%) numbers of splenic lymphocytes in nonimmunized 10-day-old rats, respectively: 0.18 ± 0.08 × 10 8 (13 ± 5) CD45 + ; 0.13 ± 0.04 × 10 8 (9 ± 3) OX12 + ; 0.07 ± 0.01 × 10 8 (5 ± 1)W3/25 + CD3 + ;0.03 ± 0.01 × 10 8 (2 ± 0.5)OX8 + CD3 + cells. Because of the difficulty in administering SRBC by tail vein injection, the pups received SRBC intraperitoneally but were still unable to generate a primary IgM response. Histologically germinal centers were observed in rat pups immunized with SRBC. Taken together,these results suggest that the spleens of 10-day-old rats consist predominantly of functionally immature lymphocytes. Analysis by flow cytometry indicated the following mean absolute and relative(%)numbers,respectively, for nonimmunized 21-day-old rats: 0.96 ± 0.25 × 10 8 (48 ± 5) CD45 + ; 0.77 ± 0.16 × 10 8 (39 ± 3) OX12 + ; 0.23 ± 0.02 × 10 8 (12 ± 2) W3/25 + CD3 + ; 0.14 ± 0.02 × 10 8 (7 ± 1)OX8 + CD3 + cells. Results when using the ELISA indicated that the SRBC-specific IgM antibody log 2 titer for rat weanlings was considerably less than the titer obtained for young adult rats, whereas results with the PFC assay indicated a response in weanlings that was within the historical range of responses for adult rats. The latter results were consistent with the histological analysis in that prominent germinal centers were observed in weanlings immunized with SRBC. Based on our flow cytometric, ELISA, and PFC assay data, a litter can be used as the unit of comparison for developmental immunotoxicity. Our results with SRBC suggest that it may not bepossible toevaluatea humoral immune functional parameter in rat pups due to the apparent immature status of their immune cells; however, additional antigens must be examined. Results in weanlings suggest that an antibody response to SRBC of sufficient magnitude can be demonstrated with the PFC assay,but that a high background may preclude the use of the SRBC-specific IgM ELISA.