应用PKA抑制法测定C1抑制剂功能活性

本文介绍了一简便、快速测定 Cl-INH 功能活性的方法。此法是向待检血浆中加入一定量的 PKA,在37℃培育一定时间后,测定残留的 PKA 活性,利用 Cl-INH 对 PKA 抑制作用来定量测定 Cl-INH 功能活性。结果表明该方法重复性好 Cl-INH 功能活性与其相应的抗原量完全相符(r=0.913,n=40)。用此法测定25份正常人血浆,18份 HAE 病人血浆,其活性分别为117.2±22.3%和20.08±10.6%。     

丙种球蛋白四种抗体国家参考品的制备

<正> 根据WHO规程对丙种球蛋白制品(简称丙球)的要求,我所于1990年开始制备了一批可检测麻疹、白喉、乙肝(抗-HBs)和甲肝抗体(抗-HAV)的丙球抗体国家参考品。该参考品原料来自上海生物制品研究所的10％人丙半成品,经分装冻干后-30℃保存。其后,用WHO的抗麻疹血清、乙肝免疫球蛋白和甲肝免疫球蛋白国际标准品进行了丙球三种抗     

丙型肝炎病毒核酸检测试剂国家参考品的制备

目的制备丙型肝炎病毒(hepatitis C virus,HCV)核酸检测试剂国家参考品。方法收集我国不同地区的献血员血浆,用核酸筛查试剂筛查HCV RNA情况,并通过PCR法对人细小病毒B19进行定性测定,筛选阳性参考品、阴性参考品和最低检出限参考品。对HCV RNA阳性样本进行基因分型;3家实验室以WHO HCV RNA国际标准品为对照品,对参考品进行协同标定;并检测参考品的稳定性和适用性。结果筛选出10份HCV阳性,而HBV、HIV、B19均为阴性的样本作为阳性参考品,其中有6份为1b型,1份为2a型,1份为3a型,1份为3b型,1份为6a型;10份HCV、HBV、HIV、B19均为阴性的样本作为阴性参考品;1份冻干的HCV RNA阳性,HBV、HIV、B19为阴性的样本作为最低检出限参考品,基因型为1b型。3个实验室经协作标定,阴性参考品的结果均为阴性,阳性参考品的HCV RNA浓度在104~105 IU/ml之间,最低检出限参考品的HCV RNA浓度为6.18×106 IU/ml。-20℃放置4周、4℃放置12 d、室温放置7 d以及反复冻融5次对参考品的HCV RNA含量无明显影响。参考品适用于目前国内市场上主要国产血液筛查和定量试剂的检测。结论制备了HCV核酸检测试剂国家参考品,可用于HCV核酸诊断试剂的质量评价和控制。     

人组织激肽释放酶基因的克隆及融合蛋白的表达

目的 开发激肽释放酶基因工程产品,为开展基因治疗高血压奠定基础。方法 采用RT-PCR的方法合成人胰腺 cDNA,从中扩增出Kallikrein(KK)基因。经 XhoI和EcoR I双酶切后,连接到pET-28b(+)载体上,酶切鉴定后,进行核苷酸序列分析和融合蛋白的表达。结果 将IPTG诱导表达的菌体进行SDS-PAGE电泳,与蛋白标准品比较,在相对分子质量31800处可见明显的高表达带。免疫印迹实验表明重组蛋白具有KK的抗原性。结论 已成功克隆并表达了人胰腺组织激肽释放酶基因,为进一步开发基因工程产品及进行基因治疗高血压研究打下了基础。     

人脲原体核酸检测试剂盒国家参考品的制备

目的制备人脲原体核酸检测试剂盒国家参考品。方法培养人脲原体两大生物群14个标准血清型菌株,培养液灭活后经颜色改变单位(color change unit,CCU)法与PCR相结合的方法进行定值,组成能覆盖脲原体所有表型及基因型的14份阳性参考品P1-P14(浓度为106 CCU/ml)及14份检测限参考品L1-L14(浓度为104 CCU/ml);选择6份能排除脲原体核酸检测试剂盒非特异性或交叉反应的样品组成阴性参考品N1-N6;选择生物群1的微小脲原体(ureaplasma parvum,UP)1和UP14作为重复性参考品R1、R2(浓度分别为105和104 CCU/ml)。用5家公司生产的脲原体核酸检测试剂盒对参考品进行验证,确定准确性、特异性、检测限、重复性、稀释线性的性能指标;并考察参考品在不同条件下(2~8℃放置7 d,37℃放置3、7、12 d,-20℃及常温反复冻融5次)的稳定性。结果支原体各培养液经CCU法测定,浓度均在106~107 CCU/ml之间,以CCU法测定浓度为靶值,绝对偏差均在±0.5个数量级以内。4家公司生产的不同试剂盒检测参考品,在准确性、特异性、检测限及重复性上均符合要求;在稀释线性上,最低稀释浓度为102 CCU/ml的样品只有2家可以检出,其他几家的稀释线性相关系数以103~106 4个浓度计算,r值均0.990 0;1家(RNA检测)检测R2的重复性,CV为5.5%。经不同条件处理的参考品的稳定性有轻微变化,但均处于设定的可接受范围内。结论制备了人脲原体核酸检测试剂盒国家参考品,该参考品可满足人脲原体核酸分型、定性及定量检测要求,经多家实验室进行验证,可用于国内大多数试剂盒的性能评价及临床实验室质量评价。     

冻干重组乙型肝炎疫苗国家参考品的制备及标定

目的 制备1批冻干参考品,用于体外相对效力测定准确评价重组(啤酒酵母)乙肝疫苗的生物学活性。方法 疫苗原液中分别加入5％和1％的蔗糖和明胶作为保护剂,制成冻干参考品。检测各项质量指标并观察其稳定性,并以Merck公司的参考品为标准,以ELA法协作标定其体外相对效力。结果 各项质量指标均符合现行规程要求,体外相对效力保持稳定,分装量准确,原液蛋白均值为24.72μg/ml,相应液体疫苗蛋白均值为12.27μg/ml。并经3家实验室协作标定,24次检定结果合并计算相对效力为0.993。结论 该冻干参考品可用作重组啤酒酵母乙肝疫苗效力检定的质控参考品,该批参考HBsAg含量定为11.0μg/ml。     

组织激肽释放酶的研究进展

组织激肽释放酶为一类性质相似的化合物,可由肝脏、胰脏、脑神经等合成。人类多种疾病与激肽释放酶基因表达相关,该基因是高血压基因治疗研究的首选靶点。本文综述了激肽释放酶基因与疾病的相关性以及该基因重组载体的构建和表达等研究进展。     

炭疽噬菌体国家参考品的研制

目的研制炭疽鉴别试验及炭疽活疫苗纯菌检查用炭疽噬菌体国家参考品。方法制备炭疽噬菌体参考品,建立噬菌体参考品效价质量标准,组织4个实验室对该参考品进行协作验证,并对参考品稳定性进行分析。结果研制的炭疽噬菌体国家参考品,其效价质量标准应不低于1.0×108PFU/ml;经协作验证,该参考品可有效鉴别3批炭疽活疫苗;该参考品在4℃条件下稳定性较好,37℃稳定性较差。结论研制的炭疽噬菌体国家参考品已被国家有关部门正式批准,可用于炭疽鉴别试验及炭疽活疫苗纯菌检查。     

冻干血吸虫病诊断试剂IgG免疫血清国家参考品的制备及标定

目的建立血吸虫病诊断试剂IgG免疫血清国家参考品。方法收集血吸虫病疫区粪检筛查阳性病人血清和非疫区正常人血清,经标定和验证合格,制成冻干参考品。结果冻干参考品血清效价符合要求,2家协作单位用3种免疫学方法标定,结果一致。结论所制备的冻干参考品可以用于血吸虫病抗体诊断试剂的检定。     

风疹减毒活疫苗病毒滴定国家参考品的制备及标定

目的制备风疹减毒活疫苗病毒滴定国家参考品,并进行标定。方法选择国内风疹减毒活疫苗生产毒株BRDⅡ制备风疹减毒活疫苗参考品,并对其进行鉴别试验、水分含量、病毒滴度及无菌检查等检定;检定合格后,组织4个实验室对候选参考品和国际参考品的病毒滴度协作标定,计算实验室内和实验室间变异系数;对候选国家参考品进行稳定性分析。结果制备的候选参考品鉴别试验、无菌检查结果均符合规定,水分含量为1.7%,病毒滴度为4.6lgCCID50/ml。经协作标定,风疹减毒活疫苗病毒滴定候选国家参考品的病毒滴度为(4.56±0.52)lgCCID50/ml,实验室内变异系数在1.25%～2.94%之间,实验室间变异系数为5.72%;国际参考品的病毒滴度为(3.96±0.08)lgCCID50/ml,实验室内变异系数在1.19%～3.21%之间,实验室间变异系数为2.04%。经考察,该参考品具有较好的稳定性。结论制备的参考品符合作为风疹减毒活疫苗病毒滴定国家参考品的要求。     

Compartmentalization Role of A-Kinase Anchoring Proteins (AKAPs) in Mediating Protein Kinase A (PKA) Signaling and Cardiomyocyte Hypertrophy

The Beta-adrenergic receptors (β-ARs) stimulation enhances contractility through protein kinase-A (PKA) substrate phosphorylation. This PKA signaling is conferred in part by PKA binding to A-kinase anchoring proteins (AKAPs). AKAPs coordinate multi-protein signaling networks that are targeted to specific intracellular locations, resulting in the localization of enzyme activity and transmitting intracellular actions of neurotransmitters and hormones to its target substrates. In particular, mAKAP (muscle-selective AKAP) has been shown to be present on the nuclear envelope of cardiomyocytes with various proteins including: PKA-regulatory subunit (RIIα), phosphodiesterase-4D3, protein phosphatase-2A, and ryanodine receptor (RyR2). Therefore, through the coordination of spatial-temporal signaling of proteins and enzymes, mAKAP controls cyclic-adenosine monophosphate (cAMP) levels very tightly and functions as a regulator of PKA-mediated substrate phosphorylation leading to changes in calcium availability and myofilament calcium sensitivity. The goal of this review is to elucidate the critical compartmentalization role of mAKAP in mediating PKA signaling and regulating cardiomyocyte hypertrophy by acting as a scaffolding protein. Based on our literature search and studying the structure–function relationship between AKAP scaffolding protein and its binding partners, we propose possible explanations for the mechanism by which mAKAP promotes cardiac hypertrophy.     

Cr^3＋=Al2（WO4）3晶体生长助熔剂体系的研究

本文用DTA和XRD法研究了Na_2O-Al_2O_3-WO_3三元系中的Al_2(WO_4)_3-Na_2WO_4截面的相平衡关系。在这截面中发现一新化合物AlNa_3(WO_4)_2。以45mol％Na_2WO_4为助熔剂生长了Cr~(3+):Al_2(WO_4)_3晶体。     

BIOALKYLATION OF BENZ(A)ANTHRACENE: IMPLICATIONS FOR CARCINOGENESIS

Previous studies on the metabolic activation of unsubstituted polycyclic aromatic hydrocarbons have largely focused on the formation of oxidation products as a result of reaction of the parent hydrocarbon with microsomal enzymes. These products of reaction have included both epoxide as well as a diol epoxide metabolites. In addition, formation of quinones as well as hydroxyl substituted derivatives of various polycyclic aromatic hydrocarbons have been investigated as metabolic endpoints. Several studies have taken place illustrating the formation of these metabolites, as well as the formation of biologically relevant adducts in a variety of animal models as well as cell culture preparations. Although these metabolic products have been widely characterized and studied in a variety of systems, there have been limited studies characterizing the formation of methyl substituted derivatives of unsubstituted aromatic hydrocarbons, and the subsequent formation of hydroxyalkyl derivatives of various compounds. The present study investigates the formation of both alkyl and hydroxyalkyl substituted derivatives of the unsubstituted polycyclic aromatic hydrocarbon benz(a)anthracene in preparations of rat liver cytosol fortified with S-adenosyl-L-methionine. The results demonstrate that methylated derivatives are formed as metabolic products, and that the identity of these products are both the 7-methybenz(a)anthracene and 7,12-dimethylbenz(a)anthracene. These results confirm that aromatic hydrocarbons undergo biological substitution reactions at specific centers of reactivity of the unsubstituted molecule yielding bioalkylated derivatives.     

一种新型不对称马来酰亚胺单体的合成

以马来酸酐 (MA) ,D ,L 氨基丙酸 (Aln)为原料合成了一种既含不对称碳原子又含官能团 (羧基 )的新型马来酰亚胺单体N (异丙酸基 ) 马来酰亚胺 (AMI)。考察了溶剂、原料配比、催化剂用量、反应时间对产率的影响 ,确定了最佳反应条件。所得N (D ,L 异丙酸基 )马来酰亚胺的产率达 52 %。并用PERKIN ELMERFTIR 1 71 0红外光谱仪、PE 2 4 0 0有机元素分析仪对产品进行了检测。本实验具有产率高、溶剂易回收、AMA不需纯化等优点。     

天蚕素A-蛙皮素杂合肽突变体的构建、表达及其抗菌活性

目的构建天蚕素A(1～8)-蛙皮素(1～12)杂合基因抗菌肽(CA-MA杂合肽)突变体,在大肠杆菌中融合表达,并进行抗菌活性检测。方法采用PCR体外定点突变技术,设计1对方向相反的引物,其中1个引物引入突变点,应用高保真的PolybestDNA多聚酶进行重组表达质粒pGEX-4T-1-CA-MA的PCR扩增,使CA-MA杂合肽第16位密码子由AGT突变为TGG。将扩增片段自身连接,构建CA-MA杂合肽突变重组表达质粒pGEX-4T-1-W16-CA-MA,转化E.coliBL21,IPTG诱导表达突变体蛋白W16-CA-MA,并对表达产物进行纯化。分离GST融合蛋白后,进行抗菌活性检测。结果重组突变表达质粒DNA测序结果表明,在预期位点发生了突变;突变蛋白在大肠杆菌中的表达量约占菌体总蛋白的18%;纯化后蛋白纯度可达75%以上,并具有一定的抗菌活性。结论已成功获得了具有抗菌活性的杂合肽突变体W16-CA-MA。     

双(苯并噁唑)芪的光物理行为及其异构化反应的研究

研究了双（苯并唑）芪的光物理行为和光顺反异构化反应,对这类化合物的发光光谱不依赖于溶剂极性的原因进行了初步讨论．异构化反应中,发现乙二醇引入时会加速该化合物反－顺异构化和抑制顺－反过程的回复,其原因可能是与生成了顺式异构体两个苯基苯并唑间的氢键有关．     

双对二甲氨基苯甲醛缩1,4-二(3-氨基丙基)哌嗪希夫碱的合成工艺及抑菌活性研究

以对二甲氨基苯甲醛(DMAB)和1,4-二(3-氨基丙基)哌嗪(BAPP)为原料,通过缩合反应合成了一个新的DMAB-BAPP希夫碱.利用正交试验确定了最佳合成工艺条件,即反应温度50℃,反应时间4h,DMAB与BAPP投料摩尔比是2.1∶1,在此条件下,产物收率为83.7％.通过琼脂扩散法测定了产物的抑菌性能,结果表...     

(E4,Z9)-TETRADECADIENAL, A SEX PHEROMONE FOR THREE NORTH AMERICAN MOTH SPECIES IN THE GENUS Saturnia

The lepidopteran genus Saturnia has three representatives in North America, S. walterorum, S. mendocino, and S. albofasciata. (E4,Z9)-Tetradecadienal (E4,Z9–14 : Ald) was identified as a sex pheromone component for all three species by combinations of coupled gas chromatography–electroantennogram detection (GC-EAD), GC–mass spectrometry (MS), and field trials. In field trials, all three species were strongly attracted to (E4,Z9–14 : Ald) as a single component. Small amounts of (Z)-9-tetradecenal (Z9–14 : Ald) also were found in extracts of all three species, but blends of this compound with E4,Z9–14 : Ald were no more attractive to male moths than E4,Z9–14 : Ald alone. Extracts of pheromone glands of female S. walterorum occasionally contained a third, trace compound eliciting responses from male antennae in GC-EAD experiments, but this compound was not identified. It is suggested that the three species can use the same, single component as a sex attractant because the flight period of S. albofasciata (fall) is different than that of the other two species (spring), whereas the geographic distributions of S. mendocino and S. walterorum overlap over only small portions of their ranges. Furthermore, the latter two species readily hybridize, so there may be minimal fitness cost to cross-attraction.     

新型FED用蓝粉Sr_5(PO_4)_3Cl: Eu~(2+)的研究

阴极射线显示器用蓝粉一直是限制彩色显示器发光效率和亮度的重要因素之一,这一问题也 同样存在于ＦＥＤ显示器件中。本文报导了最新研制的一种新型蓝色荧光粉Ｓｒ_５（ＰＯ_４）_３Ｃｌ： ＥＵ~（２＋）, 具有发光亮度较高,色纯度好,对阴极不易产生污染的特点。