HpaA IgY抑制小鼠胃内幽门螺杆菌的定植

目的 在构建基因工程菌pQE30-HpaA-DH5a的基础上,制备HpaA重组蛋白,以此为抗原,制备抗HpaA的蛋黄抗体(HpaA IgY).通过小鼠口服试验,验证HpaA IgY阻止H.pylori在胃内定植的作用,为研制预防H.pylori感染的制剂奠定基础.方法 大量诱导工程菌pQE30-HpaA-DH5a,获得重组蛋白Hpan,免疫产蛋立克体鸡,以水稀释法联合硫酸铵沉淀法提纯LgY,SDS-PAGE分析纯度,Bradford法测定浓度,Western blot鉴定对相应抗原的特异性,ELISA法检测效价.建立H.pylori感染Balb/c小鼠的动物模型,预防组在小鼠灌喂H.pylori菌液前灌喂不同剂量的HpaA LgY.组织学检查和细菌培养观察H.pylori定植.结果 提纯后的HpaA IgY纯度为90%,浓度为24.6 mg/mL,Western blotting证实有良好抗原结合特异性,ELISA效价为1:12 800.阳性对照组H.pylori的感染率为70.4%,12周后的感染率为88.9%.预防组的感染率明显低于阳性对照组,随IgY剂量的增加,感染率降低.IgY的剂量为6 mg/mL时,能达到预防效果.结论 成功制备出高纯度、高浓度、高效价的特异性HpaAIgY,在小鼠体内能阻止H.pylori的定植,有望进一步开发成为预防H.pylori感染的制剂.     

幽门螺杆菌黏附素A有效T和B联合抗原表位噬菌体展示和抗原性测定

目的 分析并确定幽门螺杆菌黏附素A(HpaA)分子中有效T细胞和B细胞联合(T/B)抗原表位.方法 重组表达HpaA(rHpaA)并免疫家兔制备抗血清.采用生物信息学技术预测和分析HpaA分子中T细胞和B细胞表位,PCB扩增T/B联合表位肽片段并构建其噬菌体展示系统.采用PEG/NaCl沉淀法提纯展示了T/B联合表位的重组噬菌体PⅢ蛋白(rPⅢ).分别以商品化幽门螺杆菌全菌IgG抗体和rHpaA抗血清为一抗,采用Western blot和ELISA对rPⅢ蛋白中展示的T/B联合表位进行筛选和鉴定.采用MTT检测rHpaA免疫小鼠脾细胞在不同重组噬菌体蛋白刺激下的增殖情况.结果 HpaA分子中共有5个T/B联合表位:HpaA10、HpaA37、HpaA79、HpaA116和HpaA143.所有T/B联合表位均成功地展示于M13噬菌体PⅢ蛋白表面.Western blot、ELISA和淋巴细胞增殖试验结果 均显示,HpaA116是优势抗原表位,HpaA37和HpaA79为有效抗原表位,HpaA10和HpaA143为无效抗原表位.结论 本研究成功地构建了幽门螺杆菌HpaA的T/B联合表位肽噬菌体展示系统.HpaA37和HpaA79,尤其是HpaA116是HpaA有效T/B联合抗原表位.     

特异性抗原幽门螺杆菌尿素酶B的体外表达

目的：建立自患者体内分离的幽门螺杆菌菌株及其抗原尿素酶B(ureB)体外表达的方法。方法：分离培养胃病患者感染的幽门螺杆菌，采用基因体外重组技术分离ureB基因，并经测序鉴定，所表达的抗原蛋白用Western blot进行鉴定。结果：测序结果证实克隆的基因与GeneBank中的序列相符，经Weslern blot证实获得ureB的重组蛋白。结论：获得了高表达的重组抗原蛋白，为ureB检测及Hp感染的诊断提供了物质基础。     

抗人蔗糖酶卵黄抗体的制备及其稳定性和体外活性研究

目的:制备抗人蔗糖酶(Sucrase,SUC)卵黄抗体(egg yolk immunoglobulin Y,IgY)并对其稳定性、体外活性进行研究。方法:采用人SUC 蛋白为抗原,免疫海兰灰蛋鸡,水稀释和盐析法提取纯化IgY;间接ELISA 法监测抗体效价变化并评价其稳定性;SDS-PAGE 和Western blot 分析IgY 的纯度和特异性;PNPG 法测IgY 对琢鄄葡萄糖苷酶的体外抑制效果。结果:初次免疫10 d 后检测到IgY,40 d 后效价最高达到1 12 800;提取的IgY 重链和轻链分别在65 kD 和25 kD 处,且能够与抗原蛋白特异性结合;29 ~69益范围内水浴15 min 和pH 4 ~7 之间37益水浴4 h,抗体效价无变化;胰蛋白酶与胃蛋白酶作用IgY2 h,抗体效价降至一半,延长作用时间,IgY 对胃蛋白酶耐受力较胰蛋白酶耐受力更强;IgY 对琢鄄葡萄糖苷酶具有一定的抑制作用,呈现剂量相关性,半抑制浓度(IC50)值为0郾540 mg/ ml。结论:抗人蔗糖酶卵黄抗体(S-IgY)的成功制备为后续用于2型糖尿病大鼠模型体内实验研究奠定基础。     

融合蛋白VacA-HpaA卵黄抗体的体外研究

目的：研究VacA-HpaA融合蛋白的鸡卵黄抗体（IgY）的理化特性和生物学活性,为制备集预防与治疗为一体的抗H.pylori感染的制剂奠定基础。方法：以纯化的VacA-HpaA融合蛋白免疫产蛋鸡,水稀释法获取蛋黄抗体（VacA-HpaAIgY）。（1）用硫酸铵沉淀法纯化IgY后,用SDS-PAGE分析其纯度,Bradford法测定蛋白含量。（2）用间接ELISA法检测该IgY的耐热性、耐酸性及其对酶消化的耐受作用。（3）采用Western blot分析IgY的特异性并检测其效价。（4）用MTT法检测不同浓度VacA-HpaAIgY对细胞毒活性的中和作用。（5）用扫描电镜观察VacA-HpaA IgY对AGS细胞粘附的中和作用。结果：（1）该IgY纯度为60%,蛋白浓度为22g/L;（2）具有一定的耐热性、耐酸性和耐胃蛋白酶能力;（3）Western blot分析显示IgY的特异性,在相对分子量约27000和30000处出现在相应的条带;（4）该IgY对VacA细胞毒活性有中和作用,且具有量效性和时效性;（5）IgY可阻止H.pylori菌体蛋白的粘附作用。结论：该IgY具有良好的理化和生物学特性,可用于制备抗H.pylori感染的防治制剂。     

特异抗内毒素鸡蛋黄免疫球蛋白效价测定及理化性质研究

目的：研究特异性抗内毒素鸡蛋黄免疫球蛋白理化性质，探索一种防治肠源性感染，治疗：内毒素血症的新途径。方法：用大肠杆菌J5株(菌体表面类脂A裸露)作抗原免疫25周龄Roman鸡，制备特异性抗内毒素IgY，用酶联免疫吸附技术检测其效价及对热、酸、碱和酶的稳定性。结果：IgY抗体效价为1：25 600，能耐受巴氏消毒，对酸、碱、酶有很好的稳定性。结论：用大肠杆菌J5株免疫鸡制备的IgY效价高，理化性质稳定，适合口服应用。     

人IL-37在大肠杆菌表达及多克隆抗体的制备与鉴定北大核心CSCD

目的原核表达白细胞介素37(IL-37)及制备其多克隆抗体。方法 PCR扩增IL-37b成熟肽编码区基因,克隆至表达载体pET28a,转化大肠杆菌感受态细胞,IPTG诱导表达,Ni2+-NTA琼脂糖凝胶柱亲和层析纯化重组蛋白,以纯化的重组蛋白IL-37为免疫原免疫BALB/c小鼠制备其特异性抗体,用ELISA、Western blot法和免疫组织化学染色检测抗体的效价和特异性。结果原核表达了重组蛋白IL-37b成熟肽,并获得高效价的小鼠抗IL-37抗体,能特异性识别天然的IL-37抗原。结论成功制备效价高、特异性好的小鼠抗IL-37抗体。     

人IL-37在大肠杆菌表达及多克隆抗体的制备与鉴定

目的原核表达白细胞介素37(IL-37)及制备其多克隆抗体。方法 PCR扩增IL-37b成熟肽编码区基因,克隆至表达载体pET28a,转化大肠杆菌感受态细胞,IPTG诱导表达,Ni2+-NTA琼脂糖凝胶柱亲和层析纯化重组蛋白,以纯化的重组蛋白IL-37为免疫原免疫BALB/c小鼠制备其特异性抗体,用ELISA、Western blot法和免疫组织化学染色检测抗体的效价和特异性。结果原核表达了重组蛋白IL-37b成熟肽,并获得高效价的小鼠抗IL-37抗体,能特异性识别天然的IL-37抗原。结论成功制备效价高、特异性好的小鼠抗IL-37抗体。     

抗HSPC238单克隆抗体的制备和初步鉴定

目的制备抗HSPC238的单克隆抗体，为HSPC238的功能研究奠定基础。方法以纯化的重组蛋白HSPC238为抗原，免疫BALB／c小鼠，运用杂交瘤技术制备HSPC238单克隆抗体，并用间接ELISA法和Western—blot法对单克隆抗体的特性进行鉴定。结果成功建立两株稳定分泌抗HSPC238的单克隆抗体杂交瘤细胞株，分别命名为E001和E002。ELISA检测抗HSPC238单克隆抗体的腹水效价为1：12800和1：25600。两株单克隆抗体的免疫球蛋白亚类均为IgG1。通过Western—blot实验证实，两株单克隆抗体均能特异性结合真核细胞内源性HSPC238蛋白。结论成功制备了两株效价高、特异性好的抗HSPC238单克隆抗体，制备的抗HSPC238单克隆抗体可用于HSPC238蛋白的鉴定，为HSPC238蛋白的生物学功能研究奠定了基础。     

抗人木糖基转移酶蛋白多克隆抗体的制备及鉴定

目的：制备兔抗人木糖基转移酶（XT-I）蛋白的多克隆抗体，并对其特异性进行鉴定。方法：应用DNAssist软件分析XT-I蛋白的抗原表位，从中选择优势抗原表位，通过引物优化设计，PCR拼接并扩增相应区域DNA片段，将其插入原核表达载体pGEX-4T-2，转化大肠杆菌ER2566，获得融合表达产物。将纯化的可溶性蛋白GST-XT作为抗原，免疫新西兰大白兔，制备抗XT-I蛋白的多克隆抗体，ELISA间接法测定效价。GST柱亲和层析法纯化抗体，采用Westem blot法鉴定该抗体的特异性和生物学活性。结果：确定XT-I的N端氨基酸序列QKHQPELAKKPPSRQKELLKRKLEQQEKGKG（175-205）为抗原表位，成功构建了重组表达载体，表达融合蛋白GST-XT，并以其为抗原，免疫新西兰大白兔，制备了抗XT-I蛋白的多克隆抗体。ELISA证实其效价高达1：640000：Western blot结果显示：该抗体可与人涎腺腺样囊性癌肺转移细胞株ACC-M中表达的XT-I蛋白特异性结合。结论：获得具有高特异性的兔抗人XT-I蛋白多克隆抗体，为涎腺肿瘤性肌上皮细胞蛋白多糖生物合成的研究提供了特异性抗体。     

Schizophrenia in a North Swedish geographical isolate, 1900–1977. Epidemiology, genetics and biochemistry

Over 200 schizophrenic patients belonging to three major and interrelated pedigree complexes have been investigated over the past 30 years in a North Swedish geographically isolated population, presently numbering about 6,000. An intensive investigation of a number of biochemical correlates and genetic markers in a few selected families belonging to one of the major pedigrees has indicated new strategies for the current research program.
Schizophrenia, as defined operationally, is significantly associated with decreased activities of two enzymes (1) blood platelet monoamine oxidase, (2) plasma dopamine-β-hydroxylase, and (3) with the genetic marker Gc² (group specific antigen). Both enzymes are subject to genetic variation. A positive score for linkage between schizophrenia and low plasma DBH activity has been calculated, but, so far, available data are insufficient for discrimination between linkage and partial contribution of genetically controlled low plasma DBH to the pathogenesis of the disease. Alternatively, both mechanisms could be involved.
As a model for continued research, schizophrenia is explained as based on a double dominant-recessive genotype (Aabb), representing a vulnerability which in about 50 % of cases develops into clinical schizophrenia. It is suggested that the dominant mutation (A) operates on or affects MAO activity, and that the recessive genotype (bb) is instrumental in low variates of DBH activity and very likely such variates within the normal range of physiological variation. Moreover, it is suggested that the combined effects of MAO- and DBH-reduced efficiency on the metabolism of e.g. dopamine could be an essential pathogenic mechanism for the schizophrenic illness which is segregating in this population.     

HLA in North Colombia Chimila Amerindians

HLA-A,-B,-C,-DRB1 and -DQB1 alleles have been studied in Chimila Amerindians from Sabana de San Angel (North Colombian Coast) by using high resolution molecular typing. A frequent extended haplotype was found:HLA-A*24:02-B*51:10-C*15:02-BRB1*04:07-DQB1*03:02 (28.7%) which has also been described in Amerinndian Mayos Mexican population (Mexico, California Gulf, Pacific Ocean). Other haplotypes had already been found in Amerindians from Mexico (Pacific and Atlantic Coast), Peru (highlands and Amazon Basin), Bolivia and North USA. A geographic pattern according to HLA allele or haplotype frequencies is lacking in Amerindians, as already known. Also, five new extended haplotypes were found in Chimila Amerindians. Their HLA-A*24:02 high frequencies characteristic is shared with aboriginal populations of Taiwan; also, HLA-C*01:02 high frequencies are found in New Zealand Maoris, New Caledonians and Kimberly Aborigines from Australia. Finally, this study may show a model of evolutionary factors acting and rising one HLA allele frequency (-A*24:02), but not in others that belong to the same or different HLA loci.     

Renal dysplasia and asplenia in two sibs

A family is reported in which two sibs, one male and the other female, both died within 24 hours of birth with enlarged polycystic kidneys. Postmortem histology in the second child showed gross renal dysplasia. In both children the pancreas was enlarged, nodular and cystic but the liver appeared macroscopically normal. In the second child, histological examination confirmed pancreatic fibrosis with cystic dilation of ducts, but showed portal fibrosis with bile duct proliferation in the liver.
This combination of findings is very reminiscent of those in a girl and her brother reported by Ivemark et al. (1959). The children reported here also showed absence or hypoplasia of the spleen, cardiac anomalies and other features of the Ivemark syndrome (Ivemark 1955), a quite different, usually sporadic, congenital disorder. It is suggested that the children described here have a distinct lethal congenital disorder, probably inherited in an autosomal recessive manner.     

The universal muscular chamber. A basic unit of the genitourinary, cardiovascular, gastro-intestinal, skeletal, cutaneous, respiratory and other systems, endocameral hypertension; and spasm, the key to their idiopathic diseases and arthropathies

Starting with the integument, we see many organs are contractile sacs or multiples thereof, which tubes or bags constitute the major part of the entire body. Recognition of this basic unit and its characteristics sheds new light, individually and collectively, on many disorders previously considered unrelated. Muscular tears and perforations develop in the walls of these chambers, being no way peculiar to those organs, wherein, hydrochloric acid occurs. So, it is not necessary to explain the absence of excessive acid from patients who exhibit holes in the gastric, uterine, aortic, duodenal, rectal, pulmonary, retina, and other walls. Muscle, not acid is the great common factor relating idiopathic disorders in the gastrointestinal tract to each other and to similar diseases in other systems. When the units are linked together, the lesions tend to appear as arthropathies, i.e. at the joints. Rephrasing common-place observations, frees us from conventional, conceptual cul-de-sacs. An observation is only as good as its interpretation, so all possibilities must be considered, otherwise, we will remain blinded by our misconceptions.     

Der Einfluß von Nahrungsmitteln und Rauchen auf die klinisch-chemische Diagnostik von Phäochromozytom,Neuroblastom und Karzinoid-Syndrom

Zusammenfassung Der Einfluß von verschiedenen Nahrungsmitteln auf Methoden zur Bestimmung von Adrenalin (AD), Noradrenalin (NA), Vanillinmandelsäure (VMS), Metanephrinen (MN), Homovanillinsäure (HVS) und 5-Hydroxyindolessigsäure (5-HIE) im 24 h-Harn zur Diagnose des Phäochromozytoms bzw. Karzinoid-Syndroms wurde untersucht. Die in die Untersuchung einbezogenen Nahrungsmittel waren: Tee, Kaffee, Mandeln, Ananas, Käse, Walnüsse, Vanillepudding, Bananen, Tomaten und Milchschokolade. Außerdem wurde der Einfluß des Zigarettenrauchens auf die Bestimmung von AD, NA, VMS und MN untersucht.Walnüsse führten zu einer starken Erhöhung der 5-HIE-Ausscheidung. Bananen erhöhten die Ausscheidung von AD, NA, VMS, MN und 5-HIE. Kaffee und Ananas bewirkten eine geringe Zunahme der MN-Werte. Rauchen von 20–30 Zigaretten/Tag beeinflußte keine der vier Variablen.Wenn die beschriebenen Methoden benutzt werden, sollte lediglich auf den Verzehr von Bananen und Walnüssen vor und während der Harnsammelperioden verzichtet werden, da die oberen Normgrenzen im Harn überschritten werden könnten. Ein Verzicht auf Kaffee und Ananas in normalen Mengen ist nicht erforderlich. Es besteht kein Anlaß, weiterhin die bisherigen umfangreichen Restriktionen der übrigen Nahrungsmittel beizubehalten.     

A comparative study of the effects of dimebon,obsidan, finoptin,and cordaron on the functional state of ischemic focus and size of necrotic zone in experimental myocardial infarction

Dimebon, an antihistamine agent, exerts a moderate antianginal effect, improving the function of ischemic focus in the myocardium and decreasing the necrotic zone in experimental myocardial infarction. Dimebon is less active than obsidan, finoptin (except for the size of the necrotic zone), and cordaron. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 122, No. 12, pp. 642–644, December, 1996     

Effects of sex steroid hormones on lipid peroxidation and glutathione antioxidant system in rat skin

Effects of estradiol and testosterone on the intensity of lipid peroxidation and contents of glutathione redox system components in the dermis and epidermis of rat skin were studied. Only estradiol induced considerable dose-dependent and tissue-specific biphasic antioxidant effects on the skin. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 128, No. 12, pp. 663–666, December, 1999     

Adhesion and growth of CaCo2 cells on surface-modified PEEK substrata

A series of surface-functionalized poly(ether ether ketone) (PEEK) films has been prepared by selective wet-chemistry; they are hydroxylated polymer (PEEK-OH) obtained by reduction, aminated polymer (PEEK-[]-NH2) prepared by coupling a diisocyanate reagent to PEEKOH (PEEK-[]-NCO) followed by hydrolysis, and carboxylated and aminocarboxylated polymers (PEEK-[]-GABA and PEEK-Lysine) resulting from the coupling of aminoacids to PEEK-[]-NCO. The aminated and carboxylated substrata promoted the adhesion and growth of CaCo2 cells in the presence of serum. Fibronectin (FN), an extra-cellular matrix protein, has been covalently fixed and/or adsorbed on various PEEK substrata, in the presence or not of a polymeric surfactant (Pluronic F68). The performances of the FN-grafted substrata (PEEK-[]-FN(1) and PEEK-[]-FN(2)) were significantly higher than those of reference substrata simply coated with FN (PEEK-OH(+FN)(1) and (2), PEEK-[]-NH2(+FN)(1) and (2)), considering the adhesion and spreading of CaCo2 cells in the absence of serum. Moreover, the stability of the adherent cells on the FN-adsorbed substrata dramatically depended on the experimental conditions applied during the PEEK coating with FN.