p21活化激酶4在非小细胞肺癌中的表达及其临床意义

目的:探讨p21活化激酶4(PAK4)在人非小细胞肺癌细胞系及人非小细胞肺癌组织中的表达及其临床意义。方法:采用Western blot及实时荧光定量PCR检测人支气管上皮(HBE)细胞、非小细胞肺癌细胞(A549、NCI-H520、NCI-H460和NCI-H596细胞)、20例新鲜非小细胞肺癌组织及相应的癌旁组织中PAK4的表达情况;采用免疫组化检测210例非小细胞肺癌组织PAK4的表达情况;采用Kaplan-Meier法评估非小细胞肺癌患者术后5年生存率;用Cox比例风险模型分析PAK4对患者预后的影响。结果:非小细胞肺癌细胞(A549、NCI-H520、NCI-H460和NCI-H596细胞)PAK4蛋白及mRNA表达均显著高于HBE细胞(P0.05);20例非小细胞肺癌组织中PAK4蛋白及mRNA表达高于癌旁组织PAK4蛋白及mRNA表达(P0.05);10例转移性非小细胞肺癌组织PAK4 mRNA表达显著高于10例原发性非小细胞肺癌组织(P0.05);免疫组化染色结果示210例非小细胞肺癌PAK4评分显著高于对应的癌旁组织。临床资料分析显示PAK4蛋白表达与非小细胞肺癌的分化程度、淋巴结转移、远处转移及临床分期有关(P0.05);PAK4高表达组患者5年生存率显著低于PAK4蛋白低表达组患者(logrank检验,P0.05),PAK4蛋白表达是非小细胞肺癌患者的独立预后因素。结论:PAK4蛋白高表达是非小细胞肺癌患者死亡的独立危险因素。     

CRKL在人非小细胞肺癌中的表达及意义

目的 CRKL(Crk-Like)基因在肺癌细胞系中存在着一定程度的扩增,我们将探讨接合物蛋白CRKL在非小细胞肺癌中的表达情况及与临床病理因素的关系。方法采用免疫组织化学和WB方法,分别检测了131例非小细胞肺癌组织和30例新鲜的非小细胞肺癌标本中CRKL的表达情况。结果 CRKL在正常支气管上皮细胞中呈阴性表达,而在肺癌组织中有44.3%(58/131)的病例存在阳性表达,WB检测发现肺癌中CRKL的表达明显高于对应的癌旁正常肺组织,CRKL蛋白在肺腺癌中的阳性表达为60.34%明显高于鳞癌(P=0.001),其表达与肺癌的低分化、高p-TNM分期(P=0.0035)、高Ki-67(P=0.0062)增殖指数和不良预后明显相关(P=0.0183)。结论 CRKL在肺癌中高表达,并与分化、分期和不良预后相关,提示CrkL蛋白可能在非小细胞肺癌的发生、发展中发挥作用。     

蛋白质磷酸酶2A癌性抑制因子在非小细胞肺癌组织中的表达及其临床意义

目的分析蛋白质磷酸酶2A癌性抑制因子在非小细胞肺癌组织中的表达及其临床意义。方法对220例非小细胞肺癌患者的癌变组织以及癌旁组织均实施免疫化学法检验,分析比较癌变组织以及癌旁组织中蛋白质磷酸酶2A癌性抑制因子(CIP2A)的表达情况。结果癌变组织的CIP2A阳性表达率(76.36%)显著高于癌旁组织(5.00%),P0.05。结论蛋白质磷酸酶2A癌性抑制因子可在非小细胞肺癌组织中高度表达,具有较为显著的临床意义。     

ADAM23、αvβ3在非小细胞肺癌中的表达及临床意义

目的研究ADAM23、αvβ3在非小细胞肺癌中的表达及其与患者临床病理特征的关系。方法应用免疫组化和RT-PCR方法检测52例非小细胞肺癌及其配对癌旁组织和8例良性病变组织中ADAM23、αvβ3的表达。结果非小细胞肺癌中ADAM23蛋白的阳性率(38.5%)低于癌旁组织(86.5%)及肺良性病变组织(87.5%)(P0.05),而αvβ3蛋白的阳性率(80.8%)高于癌旁组织(26.9%)及肺良性病变组(37.5%)(P0.05)。ADAM23蛋白在非小细胞肺癌中的表达与患者的年龄、性别、肿瘤大小以及组织学分型无关,而与分化程度、淋巴结转移和临床分期关系密切;αvβ3蛋白在非小细胞肺癌中的表达与患者的年龄、性别、组织学分型以及分化程度无关,而与肿瘤的大小、淋巴结转移和临床分期有关;RT-PCR结果显示AD-AM23mRNA在非小细胞肺癌中的表达与癌旁组织和肺良性病变组织无明显差异,而αv、β3在癌中的表达高于癌旁组织及肺良性病变组织;ADAM23和αvβ3的表达呈负相关(χ2=9.026,r=-0.417,P0.05)。结论 ADAM23在非小细胞肺癌中低表达,αvβ3表达增高,二者可能在肺癌的侵袭和转移中发挥重要作用,并且具有协同性。     

TTC25在非小细胞肺癌的表达及其临床病理意义

目的探讨TTC25在非小细胞肺癌中的表达及与临床病理参数关系。方法应用免疫组化及Western blot方法检测TTC25在100例非小细胞肺癌及癌旁正常肺组织中的表达与临床病理因素的关系。结果在正常支气管纤毛中可看到TTC25的表达,表达阳性率在非小细胞肺癌(82%)高于周围正常肺组织(25%,P0.05)。TTC25蛋白在非小细胞肺癌表达水平明显高于癌旁正常肺组织,与肿瘤Ki-67的表达呈正相关(P0.05),但与年龄、性别及TNM分期无相关性。结论 TTC25可能参与非小细胞肺癌的发生发展过程,可能是非小细胞肺癌靶向治疗的新靶点。     

人原发性肺癌组织中Rab相互作用溶酶体蛋白基因甲基化检测及其临床意义

目的 分析Rab相互作用溶酶体蛋白(RILP)在肺癌患者癌组织及肺癌细胞系中的表达及其甲基化情况,探索RILP基因的生物学功能。方法 收集重庆医科大学附属长寿人民医院88例肺癌患者的癌组织和癌旁组织标本,分别用免疫组化染色、荧光定量PCR检测RILP蛋白及mRNA表达水平。选取肺癌细胞系(H1299、A549、H358、H19936和H460)和正常肺上皮细胞系(BEAS-2B),用荧光定量PCR检测RILP mRNA表达水平。用甲基化特异性PCR检测肺癌组织及细胞中RILP的甲基化。分析肺癌患者一般临床资料与RILP甲基化的关系。用空载体或人源RILP表达载体转染A549细胞,用克隆形成实验检测细胞增殖活性。结果 肺癌组织RILP高表达率为13.64%,低于癌旁组织的76.14%(P <0.05),癌旁组织RILP mRNA表达水平高于肺癌组织(P <0.05),正常肺上皮细胞系中RILP mRNA表达水平均高于肺癌细胞系(P <0.05)。与正常肺上皮细胞系相比,RILP基因在肺癌细胞系中出现了明显的甲基化。60.23%(53/88)的肺癌患者发生RILP基因甲基...     

非小细胞肺癌中NDRG1与HIF-1α表达模式及相关性的研究

目的探讨非小细胞肺癌组织中NDRG1(N-myc下游调节因子1)与HIF-1α(缺氧诱导因子1α)的表达模式及关联。方法应用免疫组织化学方法检测105例非小细胞肺癌及癌旁肺组织中NDRG1和HIF-1α的蛋白表达。应用Western Blotting检测12对新鲜肺癌组织及癌旁肺组织中NDRG1及HIF-1α的蛋白表达。结果 NDRG1与HIF-1α在非小细胞肺癌组织中具有相似的表达模式,均主要表达于细胞浆,阳性率分别为55.2%(58/105)及50.5%(53/105),部分病例伴有细胞核(分别为20.0%(21/105)及35.2%(37/105))和细胞膜(分别为11.4%(12/105)及7.6%(8/105))的表达,总的阳性率分别为60.0%(63/105)及53.3%(56/105)。NDRG1在癌旁肺组织中的表达(17.1%(18/105))低于癌组织(<0.05),HIF-1α在癌旁肺组织中的表达(46.7%(49/105))与癌组织中的表达无显著差异(>0.05),NDRG1与HIF-1α在非小细胞肺癌组织中的表达水平呈正相关(r=0.210,<0.05)。Western Blotting结果显示NDRG1在肺癌组织中的表达高于癌旁肺组织(<0.05),其在癌组织中的表达与HIF-1α的蛋白表达呈正相关(<0.05)。HIF-1α在癌及癌旁组织中的表达无明显差异(<0.05)。结论 NDRG1在非小细胞肺癌高表达与HIF-1α表达呈正相关,二者可能在肿瘤内部缺氧诱导的应激反应过程中存在相互作用。     

长链非编码RNAuc.339在非小细胞肺癌中的表达及临床意义

目的：分析uc.339在非小细胞肺癌中的表达及其与临床病理特征的关系。方法：非小细胞肺癌新鲜标本及其癌旁肺组织各30例,提取总RNA,采用实时定量PCR方法检测uc.339在非小细胞肺癌组织及其癌旁肺组织中的表达,分析差异性及其与临床病理特征的关系。癌性胸腔积液及非癌性胸腔积液各20例,离心沉淀,提取总RNA,采用实时定量PCR方法检测uc.339在癌性胸腔积液及非癌性胸腔积液中的表达;免疫组织化学检测P53在非小细胞肺癌中的表达。结果：uc.339在非小细胞肺癌组织中的表达低于癌旁肺组织(P<0.05),与临床分期有相关性(P<0.05),与p53的表达呈负相关(P<0.05),而与肿瘤病理类型、年龄、性别无明显相关性(P>0.05);uc.339在癌性胸腔积液中的表达低于非癌性胸腔积液(P<0.05)。结论：uc.339可能作为一种抑癌因子作用于非小细胞肺癌的发生、发展中,其作用机制可能与p53信号通路有关,在作为肿瘤标志物、判断预后及靶向治疗等方面具有应用价值。     

TrkB/BDNF在非小细胞肺癌组织中的表达及临床意义

目的:探讨肿瘤标记物TrkB/BDNF在非小细胞肺癌癌组织中的表达及其临床意义。方法:采用免疫组化(SP法)和Realtime-PCR方法检测非小细胞肺癌组织和癌旁组织中TrkB/BDNF的mRNA表达水平,并分析其在相关临床因素间的差异。结果:非小细胞肺癌癌组织TrkB/BDNF表达阳性,且明显高于癌旁组织。TrkB/BDNFmRNA水平在有否脑转移和不同TNM分期之间存在明显差异(P<0.05);而在性别、年龄、吸烟、肿瘤大小、分化程度、病理组织学类型间无显著差异(P>0.05)。结论:TrkB/BDNF在非小细胞肺癌癌组织中高表达,并与其脑转移、TNM分期有关,两者可能在非小细胞肺癌的侵袭及转移(尤其脑转移)中发挥重要作用。     

PIK3CA基因对人非小细胞肺癌A549细胞侵袭及迁移能力的影响

目的探讨PIK3CA基因对非小细胞肺癌侵袭及迁移能力的影响及可能机制。方法实时荧光定量PCR检测非小细胞肺癌组织、癌旁组织、非小细胞肺癌A549细胞与人支气管上皮细胞PIK3CA mRNA的表达,构建靶向PIK3CA基因的si RNA质粒,并转染至非小细胞肺癌A549细胞,实时荧光定量PCR技术与Westem blot方法分别检测PIK3CA mRNA与蛋白表达的变化,利用Transwell实验检测转染后细胞侵袭和转移能力的变化,Westem blot方法检测转染后A549细胞p-Akt蛋白表达变化。结果与癌旁正常组织比较,PIK3CA mRNA和蛋白表达水平在非小细胞肺癌组织显著上升(P0.05),A549细胞中PIK3CA mRNA和蛋白表达水平明显高于人支气管上皮细胞(P0.05)。PIK3CA基因沉默6h,A549细胞PIK3CA mRNA和蛋白表达水明显下降(P0.05);PIK3CA基因沉默48h,A549细胞侵袭和转移能力显著降低,p-Akt蛋白表达显著降低(P0.05)。结论 PIK3CA基因能够降低非小细胞肺癌侵袭及迁移能力,其作用机制可能与调控p-Akt蛋白表达有关。     

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.     

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.     

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.     

What will studies of Fulani individuals naturally exposed to malaria teach us about protective immunity to malaria?

There are an estimated over 200 million yearly cases of malaria worldwide. Despite concerted international effort to combat the disease, it still causes approximately half a million deaths every year, the majority of which are young children with Plasmodium falciparum infection in sub-Saharan Africa. Successes are largely attributed to malaria prevention strategies, such as insecticide-treated mosquito nets and indoor spraying, as well as improved access to existing treatments. One important hurdle to new approaches for the treatment and prevention of malaria is our limited understanding of the biology of Plasmodium infection and its complex interaction with the immune system of its human host. Therefore, the elimination of malaria in Africa not only relies on existing tools to reduce malaria burden, but also requires fundamental research to develop innovative approaches. Here, we summarize our discoveries from investigations of ethnic groups of West Africa who have different susceptibility to malaria.     

'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.     

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.     

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.     

The case for allele‐specific recognition by the TCR

There is a sharp difference in how one views TCR structure–function–behaviour dependent on whether its recognition of major histocompatibility complex‐encoded restriction elements (R) is germline selected or somatically generated. The generally accepted or Standard model is built on the assumption that recognition of R is by the V regions of the αβ TCR, which is not driven by allele specificity, whereas the competing model posits that recognition of R is allele‐specific. The establishing of allele‐specific recognition of R by the TCR would rule out the Standard model and clear the road to a consideration of a competing construct, the Tritope model. Here, the case for allele‐specific recognition (germline selected) is detailed making it obvious that the Standard model is untenable.