表达CRT-E2融合蛋白肿瘤疫苗诱导特异性抗肿瘤免疫应答

目的:探讨表达钙网蛋白(Calreticulin,CRT)和HPV E2融合蛋白的肿瘤疫苗在小鼠体内诱导的抗肿瘤免疫应答.方法:转染重组质粒得到高表达CRT、E2和CRT-E2融合蛋白的肿瘤细胞,作为肿瘤疫苗隔周两次腹腔注射免疫小鼠,17天后观察成瘤率,检测NK细胞杀伤活性、特异性T细胞增殖能力、CIL活性以及睥淋巴细胞分泌IFN-γ水平,并观察荷瘤小鼠生存期.结果:高表达CRT-E2融合蛋白肿瘤疫苗免疫小鼠后,其成瘤率明显低于其他实验组,NK细胞杀伤活性、特异性T细胞增殖能力、CTL活性和脾淋巴细胞分泌IFN-γ水平均显著高于其它实验组(P＜0.01),生存期也明显延长(P＜0.01).结论:小鼠体内实验显示,表达CRT-E2融合蛋白肿瘤疫苗能够诱导特异性CD8+T细胞免疫应答和NK细胞活性,显著抑制了肿瘤生长.     

肺癌DC融合细胞诱导的抗肿瘤免疫应答

目的 研究肺癌DC融合细胞FLD－A11体内外诱导免疫应答的能力,方法 用MTT法测定FLD－A11细胞诱导淋巴细胞增殖的能力;ELISA法检测其诱导淋巴细胞分泌IL－2的水平;LDH释放法测定FLD－A11细胞免疫后小鼠脾淋巴细胞的特异性CTL活性。结果 FLD－A11有有效刺激淋巴细胞的增殖反应（SC：RC＝1：50时,SI＝2.38）,诱导淋巴细胞分泌IL－2。FLD－A11细胞免疫后,小鼠的胸腺和脾脏重量均高于对照组（P＜0.05）,脾淋巴细胞对Lewis肺癌细胞的杀伤活性显著高于对照组。结论 FLD－A11细胞具有诱导初次抗肿瘤免疫应答的能力,不仅能在体外诱导淋巴细胞的增殖和IL－2的分泌,而且体内免疫接种也能引起小鼠免疫器官的增生反应,产生针对Lewis肺癌的特异性CTL杀伤作用。     

小鼠凋亡相关新基因PNAS-4真核表达载体的构建、表达及体外抗肿瘤作用

目的：对小鼠凋亡相关新基因PNAS-4（mPNAS-4）进行克隆，构建其真核表达载体，并在小鼠Lewis肺癌细胞系LL2中转染表达；探讨mPNAS-4基因转染LL2细胞过表达所诱导的体外肿瘤细胞凋亡情况。方法：用RT-PCR从小鼠肝脏组织中克隆mPNAS-4编码区cDNA，构建重组真核表达载体pcDNA3．1（＋）-mPNAS-4；用脂质体将pcDNA3．1（＋）-mPNAS-4转染小鼠Lewis肺癌LL2细胞；用RT—PCR检测转染细胞中mPNAS-4的过表达情况；通过MTT、流式细胞术（FCM）及DNA Ladder分别检测转染细胞的增殖与凋亡情况。结果：从小鼠肝脏组织中克隆到mPNAS-4全长cDNA并成功构建其真核表达载体pcDNA3．1（＋）-mPNAS-4，转染小鼠Lewis肺癌LL2细胞可使其mRNA表达明显上调。mPNAS-4过表达能抑制LL2细胞增殖并诱导其凋亡。结论：mPNAS-4过表达对小鼠Lewis肺癌LL2细胞的生长有明显的抑制和诱导凋亡作用。     

B7.1基因的表达对肝癌细胞生物学特性的影响

目的　观察B7 1基因表达对肝癌细胞增殖特性的影响及诱发杀伤性T淋巴细胞 (CTL)细胞毒活性的变化。方法　用细胞计数法和流式细胞仪 (FCM)测定细胞生长曲线、DNA含量及细胞周期的变化。同时测定CTL对转染B7 1基因前后肝癌细胞的细胞毒作用。结果　转染B7 1基因后 ,肝癌细胞的增殖受到一定影响 :生长延缓、增殖幅度降低和倍增时间延长。CTL对转染B7 1基因的肝癌细胞杀伤明显增强。结论　转染B7 1基因的肝癌细胞出现了一定的增殖抑制现象 ,提示B7 1基因有一定的免疫调节作用     

表达HLA—G1的K562细胞抵抗外周血NK细胞杀伤作用的研究

目的 研究HLA－G1分子对NK细胞杀伤活性的影响。方法 供助脂质体介导的DNA转染技术,将本室构建的真核南闰pcDNA3-HLA-G1转染人K562细胞;通过G418筛选,获得克隆化细胞株K562－G1;应用RT－PCR及流式细胞术分别在RNA水平和蛋白质水平检测HLA－G1的表达;最后,应用MTT比色法检测转染细胞对不同个体外周血NK细胞杀伤活性的抑制效应。结果 与转染了空质粒的对照组相比,外周血NK细胞对K562－G1的杀伤率降低32.43%（P＜0.01）。结论 靶细胞表达HLA－G2分子可明显抑制NK细胞的杀伤效应。     

WWOX基因在肺腺癌中表达及对A549细胞增殖和凋亡的影响

目的研究WWOX基因在肺腺癌组织的表达及在肺腺癌细胞系A549细胞增殖和凋亡的作用,探讨肺腺癌与临床病理特征之间的关系。方法应用免疫组化技术检测62例肺腺癌组织WWOX蛋白的表达;将携有WWOX基因的表达载体转染肺腺癌细胞系A549细胞,用Western blot法检测A549细胞WWOX蛋白的表达情况;运用细胞计数法检测细胞的增殖情况;流式细胞仪观察其对A549细胞凋亡的影响。结果肺腺癌组中WWOX的阳性表达率为45.2%,明显低于癌旁正常组织(82.9%,=0.000)。WWOX在肺腺癌的表达与分化程度(=0.023)、TNM分期(=0.007)、远处转移有关(=0.009),与患者的性别、年龄以及肿瘤的大小无关(0.05)。A549细胞转染WWOX基因后,WWOX蛋白表达明显高于空载质粒组及未转染组(未检测到WWOX蛋白的表达);生长曲线可见pcDNA3.0-WWOX转染组的A549细胞的增殖速度比pcDNA3.0组和未转染组明显下降;流式细胞仪分析表明pcDNA3.0-WWOX转染组的凋亡率明显高于pcDNA3.0空载体组。结论 WWOX的表达能够抑制肿瘤细胞的增殖,促进其凋亡。     

TCRVβ8.4 基因转染人PBMC对肝癌细胞杀伤活性的研究

目的：探讨转染hTCRVβ8.4基因后健康人PBMC的免疫学特性及其对起源于乙肝病毒的肝癌细胞株BEL-7402杀伤活性的改变。方法：将hTCRVβ8.4基因克隆至真核表达载体pcDNA3．1(-)上，并转染健康人PBMC，流式细胞术检测转染后PBMC中TCRVβ8.4蛋白表达，乳酸脱氢酶释放活性法检测重组质粒转染后PBMC对癌细胞BEL-7402的杀伤活性。结果：TCRVβ8.4在基因转染后淋巴细胞中表达显著增高；与BEL-7402共培养后，基因转染组CD3^ TCRVβ8.4T细胞增殖明显高于对照组；BEL-7402刺激后免疫细胞活化，表达CD122的细胞数量增多，表达CD19(B细胞活化的标志)的B细胞增加；重组质粒转染后，PBMC对肝癌细胞BEL-7402杀伤活性增强；透射电镜观察发现，重组质粒转染的PBMC使BEL-7402凋亡。结论：hTCRVβ8.4基因修饰可显著增强淋巴细胞在超抗原BEL-7402刺激下的增殖及免疫细胞活化，基因修饰后T淋巴细胞杀伤活性明显增强。     

龙牙楤木多糖抗肿瘤活性及对荷瘤小鼠免疫功能的影响

目的:探讨龙牙楤木多糖(AEPS)抗肿瘤活性及对荷瘤小鼠免疫功能的影响。方法:以S180肉瘤为肿瘤模型,检测龙牙楤木多糖对肿瘤生长的抑制活性;MTT法检测龙牙楤木多糖对S180肉瘤细胞、A549肺癌细胞、SMMC-7721肝癌细胞的体外抑制活性;以其对荷瘤小鼠免疫器官、血液淋巴细胞数量及淋巴细胞增殖影响、巨噬细胞活性和NK细胞杀伤活性来评价AEPS对荷瘤小鼠免疫功能的作用。结果:AEPS对S180肉瘤生长有显著的抑制作用,其中75 mg/(kg.d)剂量组抑瘤率最高达57.68%;AEPS对S180肉瘤细胞、A549肺癌细胞、SMMC-7721肝癌细胞生长的最高抑制率均达60%以上;AEPS显著提高荷瘤小鼠脾脏和胸腺质量以及血液淋巴细胞数量,促进淋巴细胞增殖反应,增加NK细胞杀伤活性和巨噬细胞活性。结论:AEPS有显著的抗肿瘤活性,并能直接作用于肿瘤细胞,抑制肿瘤生长,其抑瘤作用与机体免疫功能的增强有关。     

mdr1基因转染的CIK细胞耐药性及杀伤活性的研究

目的:将多药耐药基因(mdr1)转入CIK细胞,观察转染前后其对紫杉醇的耐药性及对Lewis肺癌细胞杀伤活性的影响。方法:常规方法:培养CIK细胞,在细胞对数生长期,将mdr1的重组质粒转染CIK,通过RT-PCR鉴定耐药基因表达;MTT法检测CIK细胞对紫杉醇敏感性的变化,同时检测转染前后CIK细胞对Lewis肺癌细胞的杀伤活性变化;Western blot检测细胞中mdr1编码的P-gp蛋白的表达的变化;通过计算瘤重抑制率(TWI)检测转染前后的CIK细胞对Lewis肺癌移植瘤的抑制作用。结果:转染mdr1后的CIK细胞mdr1 mRNA阳性,同时P-gp的表达较转染前及转染空质粒的CIK细胞均有显著增高(P<0.05),转染后的CIK细胞对紫杉醇的耐药性有显著提高,但对Lewis肺癌细胞的杀伤活性无明显变化(P>0.05)。结论:将mdr1基因转入CIK细胞后,细胞获得了多药耐药性,同时保持了原有的对肿瘤细胞的杀伤活性。     

抗CD94单抗与NK细胞、γδ^＋T细胞及CTL功能的关联

目的：探讨抗CD94单抗与NK细胞、γδ^ T细胞及CTL的相关性。方法,采用荧光检测法,分别分析NK细胞、γδ^ 细胞和CTL上CD94分子的表达,采用^3H TdR掺入法和MTT释放法,分析抗94单抗对这3种细胞的杀伤活性和细胞增殖的影响。结果NK细胞和γδ^ T细胞上CD94分子的表达,明显高于PBL和CTL。抗CD94单抗能激活NK细胞、γδ^ T细胞和CTL的增殖,并能提高γδ^ T细胞和CTL的杀伤活性。结论CD94分子在NK细胞和γδ^ T细胞表面呈高表达。抗CD94单抗对NK细胞、γδ^ T细胞及CTL的增殖具有上调节作用,对它们的杀伤活性具有正向调节作用。     

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.     

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 dominant diseases of modernized societies as omega-3 essential fatty acid deficiency syndrome: Substrate beriberi

About 1900, modern food selection and processing caused widespread $\text{epidemics}$ of the B vitamin deficiency diseases of beriberi and pellagra which, for genetic reasons, often expressed as different diseases ranging from bowel and heart disease to dermatoses and psychoses. But the B vitamins merely help convert essential fatty acids (EFA) into the prostaglandin (PG) tissue regulators and it now turns out that, through hydrogenation, milling and selection of w3-poor southern foods, we have also been systematically depleting, by as much as 90%, a newly discovered trace Nordic EFA (w3) of special importance to primates and sole precursor of the PG3(4) series, even as a concurrent fiber deficiency increases body demand for EFA. Since substrate EFA is processed by many B vitamin catalysts, an EFA deficiency will mimic a $\text{panh}$ypovitaminosis B, i.e., a mixture of $\text{substrate}$ beriberi and $\text{substrate}$ pellagra resembling vitamin beriberi and pellagra but exhibiting as even more diverse $\text{endemic}$ disease. This would consitute a second stage of the Modern Malnutrition and explain why some workers now hold the dominant diseases of modermized societies to be new, nutritionally based, pellagraform yet lipid-related and to range, once again, from heart disease to psychosis. It is an assumption that our dominant diseases are unrelated to each other or are merely revealed by our diagnostic acumen and therapeutic success; and that hydrogenating millions of tons of food oils annually, to destroy the rancidity producing w3-EFA, is safe for $\text{primates}$. Extensive beriberiform disease is reported here in 32 typical cases taken from medical practice which responds strikingly to linseed oil supplements (60% w3-EFA) in confirmation of identical results in Capuchins.     

'Very sore nights and days': the child's experience of illness in early modern England, c.1580-1720

Sick children were ubiquitous in early modern England, and yet they have received very little attention from historians. Taking the elusive perspective of the child, this article explores the physical, emotional, and spiritual experience of illness in England between approximately 1580 and 1720. What was it like being ill and suffering pain? How did the young respond emotionally to the anticipation of death? It is argued that children’s experiences were characterised by profound ambivalence: illness could be terrifying and distressing, but also a source of emotional and spiritual fulfilment and joy. This interpretation challenges the common assumption amongst medical historians that the experiences of early modern patients were utterly miserable. It also sheds light on children’s emotional feelings for their parents, a subject often overlooked in the historiography of childhood. The primary sources used in this article include diaries, autobiographies, letters, the biographies of pious children, printed possession cases, doctors’ casebooks, and theological treatises concerning the afterlife.     

Guidelines for the Validation and Use of Immunoassays for Determination of Introduced Proteins in Biotechnology Enhanced Crops and Derived Food Ingredients

Recent advancements in agricultural biotechnology have created a need for analytical techniques to determine introduced proteins in crops enhanced through modern biotechnology techniques. These proteins are expressed in plant tissues and may be present in food ingredients. Immunoassays are ideally suited for protein detection and may be used as both quantitative and threshold methods. Microplate ELISA and lateral flow devices are two of the most commonly used immunoassay formats for agricultural biotechnology applications. This paper provides general background information and a discussion of criteria for the validation and application of immunochemical methods to the analysis of proteins introduced into plants and food ingredients using biotechnology methods. It is the result of a collaborative effort of members of the Analytical Environmental Immunochemical Consortium. This collaborative effort represents the combined expertise of several organizations to reach consensus on establishing guidelines for the validation and use of immunoassays. Further, the paper offers developers and users a consistent approach to adopting the technology as well as aid in producing accurate and meaningful results.     

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.     

Ultracryotomy: development and application (with examples from the yeast cytology) (author's transl)

The preparation steps usually necessary for obtaining ultrathin frozen sections of biological material (chemical prefixation, enclosing, cryoprotective treatment, freezing, sectioning, and post-staining the sections for transmission electron microscopy) are submitted to a critical analysis. The application of cryo-ultramicrotomy, in particularly for cytochemical purposes, is reviewed. Fundamental considerations of chemical prefixation and poststaining are supported by examples from yeast cytology. Furthermore, the efficiency of the cryo-ultramicrotomy (electron optical resolution of ultrastructural details) is demonstrated on yeast cells and protoplasts.     

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.