青海高原藏族胃癌患者自然杀伤细胞活化性受体D及其配体MICA表达的研究

目的： 研究高原藏族胃癌患者外周血NK细胞表面活化性受体D(NKG2D)的表达及局部肿瘤组织和癌旁正常组织中相应配体MICA 的表达,探讨宿主NKG2D 在抗胃癌中的作用及其与肿瘤免疫逃逸的关系。方法: 对33例高原藏族胃癌患者及20 例健康人,采用流式细胞术检测外周血NKG2D 的表达状况,RT-PCR 和免疫组织化学技术检测在局部肿瘤组织和癌旁正常组织标本中MICA 的表达。结果: 高原藏族胃癌患者外周血NKG2D 的表达为(13.47 ±5.26)%，明显低于正常组（32.62±10.08)%，2组之间差异显著(P<0．05); 藏族胃癌组织MICA mRNA的表达水平显著高于癌旁正常组织（P<0.05）。胃癌组织MICA蛋白表达在癌旁正常组织中为(21.21%,7/33),胃癌组织阳性率为(78.79%，26/33),高分化组(60.00%,3/5),中分化组(72.73%,8/11),低分化组(88.24%,15/17),组间差异显著(P<0.05),而与肿瘤的大小、性别、年龄、淋巴结转移无关（P>0.05）。 Spearman相关分析显示，MICA表达水平与mRNA的表达水平显著正相关（r=0.903,P<0.01）。结论: 高原藏族胃癌的免疫逃逸可能与NKG2D 表达下调及其配体MICA 的表达升高有关，高原藏族胃癌患者外周血NK细胞活性降低,其NKG2D 表达的下降是NK细胞活性下降的原因之一。NKG2D配体MICA 的基因表达可能与高原胃癌患者的恶性转化有一定的相关性。     

妊娠期uNK、pNK细胞NKG2A与NKG2D的表达及其生物学意义的研究

目的:研究妊娠期妇女子宫NK细胞(uNK细胞)与外周血NK细胞(pNK细胞)表面NKG2A和NKG2D及其相应配体的表达,探讨uNK细胞表面NKG2A和NKG2D的不平衡表达与母胎界面所形成的免疫耐受关系。方法:采用流式细胞术检测30例孕6～9周的正常妊娠妇女uNK细胞和pNK细胞NKG2A、NKG2D的表达状况;RTPCR技术检测绒毛膜组织HLAE、MICA的表达。结果:子宫NK细胞NKG2A的表达显著高于外周血NK细胞,二者分别为(97.86±1.75)%与(33.35±10.92)%;子宫NK细胞NKG2D的表达水平与外周血NK细胞相近,分别为(93.21±4.52)%与(97.80±1.72)%,滋养层组织仅检测到HLAEmRNA的表达。结论:妊娠期子宫NK细胞表面高表达抑制性受体NKG2A,同时滋养层组织表达相应的配体HLAE,这可能是维持母胎界面免疫耐受的重要因素。     

卵巢肿瘤患者外周血NK细胞受体NKG2A、NKG2D及NK活性检测的临床意义

通过研究卵巢癌及良性卵巢肿瘤患者外周血NK细胞表面受体的表达情况及NK活性的变化,分析探讨宿主NK细胞受体与肿瘤免疫逃逸的关系及其临床价值。分离受检者外周血单个核细胞,应用MTT法检测NK细胞的细胞毒活性,流式细胞术检测NK细胞受体NKG2D和NKG2A的表达,并结合临床病理因素作比较分析。结果显示,与良性卵巢肿瘤组和正常组相比,卵巢癌患者外周血NK细胞的细胞毒活性降低,NK细胞表面NKG2D的表达水平降低,而NKG2A的表达水平明显升高,其变化与卵巢癌的病情进展有关。此结果表明,卵巢癌患者机体NK细胞杀伤活性下降,NKG2D与NKG2A二者之间的平衡表达可能对NK细胞的功能状态起着重要的调节作用。     

妊娠子宫微环境中uNK细胞NKG2A和NKG2D及其配体表达的研究

目的：研究妊娠子宫微环境中子宫自然杀伤细胞（uNK细胞）NKG2A和NKG2D及其相应配体的表达，探讨NKG2A与NKG2D的不平衡表达在母胎免疫耐受形成中的作用。方法：选择30例孕6-9周的正常妊娠妇女,分离其新鲜蜕膜组织，除去绒毛,分离蜕膜和外周血单个核细胞，采用流式细胞仪测定NK细胞的数量及NKG2A与NKG2D的表达；采用RT-PCR技术检测滋养层组织NKG2A与NKG2D配体人类白细胞抗原-E(HLA-E)、主要组织相容性复合体-Ⅰ类分子相关蛋白A(MICA)mRNA的表达结果：妊娠子宫蜕膜淋巴细胞中NK细胞约占70%,流式细胞分析的结果显示，子宫自然杀伤细胞NKG2A的表达显著高于外周血NK细胞，分别为97.86%±1.75%与33.35%±10.92%（〖AKx-D〗±s），两者差异显著（P<0.05），在滋养层细胞中检测到其配体HLA-E的表达；而与外周血相比，uNK细胞表面NKG2D的表达与之较为相近，分别为93.21%±4.52%与97.80%±1.72%，但两者仍有显著差异（P<0.05）。在滋养层组织未检测到其相应配体MICA mRNA的表达结论：蜕膜中的淋巴细胞主要为NK细胞，其免疫学表型与外周血NK细胞有较大的区别，妊娠期子宫自然杀伤细胞表面高表达抑制性受体NKG2A，同时滋养层组织表达相应的配体人类白细胞抗原-E，这可能是维持母胎界面免疫耐受的重要因素。     

膜型/分泌型MICA对NK细胞受体NKG2D的相反调节效应及其对NK细胞受体谱的影响

目的　观察膜型和分泌型MICA对NK细胞受体表达的影响 ,以探讨NK细胞抗肿瘤活化机制及肿瘤细胞表达MICA分子的意义。方法　用MTT法测定人NK细胞系 (NK92 )的细胞毒活性 ;用RT PCR或FACS检测NK细胞受体 (NKG2D ,NKG2A B ,KIR2DL1,KIR2DS1)及NKG2D的识别配体MICA的表达。结果　肿瘤细胞表面的MICA分子可上调NKG2D的表达 ,下调抑制性受体NKG2A B和KIR2DL1的表达 ;而分泌型MICA (sMICA)分子对NKG2D及抑制性受体的表达均有抑制作用。结论　膜型MICA分子可上调NKG2D的表达 ,激发NK细胞对肿瘤细胞的细胞毒效应 ;分泌型MICA分子则通过降低NKG2D的表达下调机体的抗肿瘤免疫效应 ,肿瘤细胞分泌sMICA分子为肿瘤发生免疫逃逸的机制之一。     

白血病患者NK细胞表面NKG2D受体及其配体MICA/B的研究

目的:研究白血病患者NK细胞表面NKG2D受体及其配体MICA/B的表达,探讨白血病细胞逃逸NK细胞杀伤的机制.方法:采用流式细胞术检测NK细胞表面NKG2D受体和骨髓有核细胞表面MICA/B配体.结果:治疗前组和完全缓解组NKG2D受体的表达均较健康组低(P＜0.05);且完全缓解组低于治疗前组(P＜0.05);治疗前组和完全缓解组MICA/B配体的表达均低于增生性贫血组(P＞0.50);完全缓解组与治疗前组比较差异无显著性(P＞0.05).结论:白血病患者体内NKG2D-MICA/B介导的NK细胞功能受抑,这可能导致白血病细胞逃逸NK细胞的细胞毒作用;白血病化疗后完全缓解时其体内NKG2D-MICA/B介导的NK细胞功能仍未恢复,且较治疗前更低.     

人MICA基因多态性和表达特性与肿瘤的关系

近年来,关于MICA与肿瘤关系的研究进展显著。MICA是NK细胞活化性受体NKG2D的配体,可高表达于上皮源性的肿瘤细胞;NKG2D不仅表达于NK细胞,还表达于CTL和Vδ1γδT细胞;NKG2D以一种可诱导的方式识别并结合MICA,从而活化上述抗瘤效应细胞。     

5-氟尿嘧啶增强HeLa细胞对自然杀伤细胞敏感性研究

目的 用5-氟尿嘧啶(5-fluorouracil,5-FU)处理HeLa细胞,检测其NKG2D 配体 MICA 的表达及其对NK92细胞杀伤敏感性的变化.方法 不同浓度的 5-FU处理 HeLa细胞,在不同时间点用半定量PCR及流式细胞术检测HeLa细胞表面的NKG2D配体MICA在RNA及蛋白水平的表达变化情况,用MTT法检测NKG2D抗体封闭NK92细胞的NKG2D受体前后, NK92 细胞对HeLa细胞的杀伤作用.结果 不同浓度的5-FU作用于 HeLa 细胞后,半定量RT-PCR结果显示MICA表达随5-FU作用浓度增加逐渐增高.而且40 μg/ml 5-FU作用于HeLa细胞后随着作用时间的延长(0、8、16、24 h)MICA表达增加,流式细胞术检测结果表明,MICA表达的增加主要依赖于未凋亡细胞的 MICA表达.在40μg/ml 5-FU作用24 h,效靶比为2.5∶ 1,5∶ 1,10∶ 1,20∶ 1时都检测到NK92细胞对HeLa细胞的杀伤增强,杀伤作用可部分被NKG2D抗体抑制.结论 5-FU 能够上调HeLa细胞表面NKG2D配体MICA的表达,增强HeLa细胞对NK92细胞的敏感性,提示化疗联合NK细胞免疫治疗宫颈癌可产生协同作用,提高治疗效果.     

MICA分子在NK细胞杀伤乳腺癌中的作用

目的 观察乳腺肿瘤细胞膜MHC Ⅰ链相关分子A(MHC class Ⅰ chain-related gene A,MICA)及血清中可溶性MICA表达,探讨MICA在NK细胞杀伤乳腺癌中的作用.方法 流式细胞术检测膜型MICA(mMICA)及NK细胞表面NKG2D表达,观察膜型MICA、可溶性MICA(sMIcA)对NKG2D表达的影响;免疫组织化学检测膜型MICA及可溶性MICA的表达及分布;抗体封闭法观察膜型MICA与NKG2D相互作用.结果 乳腺细胞膜型MICA在正常组织不表达,在良性肿瘤表达量为(38.5±7.5)%,恶性肿瘤表达量为(53.2±5.6)%.可溶性MICA含量在健康成年人为阴性,在乳腺良性肿瘤患者血清中含量(76.8±22.3)pg/mL,在恶性肿瘤患者中含量(205.36±71.27)pg/mL.经NKG2D或MICA抗体封闭后,NK细胞的杀瘤活性显著减弱.含可溶性MICA的血清可明显下调NKG2D的表达.结论 大部分乳腺肿瘤细胞膜都有MICA的表达,可作为乳腺肿瘤相关性抗原.膜型MICA与NKG2D的相互识别在介导NK细胞抗乳腺癌中起重要作用,而可溶性MICA通过下调NKG2D表达介导肿瘤免疫逃逸.     

结肠癌患者单核或树突状细胞MICA的表达及其与NK细胞活性的关联

目的:观察结肠癌患者单核或树突状细胞是否表达MHCⅠ类相关抗原A(MICA),并分析MICA+单核细胞的频率是否与患者体内NK细胞的活性相关联。方法:首先利用流式细胞仪检测MICA在外周血单核细胞和由单核细胞诱导而来树突状细胞(DCs)上的表达;并用IL-15和IFNα-刺激树突状细胞,观察MICA表达的变化;然后检测患者外周血NKG2D+、CD16+、CD69+NK细胞的频率及NK细胞的杀伤活性;最后分析MICA+单核细胞的频率是否与NK细胞表达NKG2D及其杀伤活性关联。结果:与健康个体相比,结肠癌患者MICA+单核细胞的频率无显著变化。未成熟DCs表达MICA,但受IL-15和IFNα-刺激后,结肠癌来源的DCs表面MICA的表达无显著上调。结肠癌患者外周血NKG2D+、CD69+NK细胞的频率显著降低,NK细胞的杀伤活性显著降低,而CD16+NK细胞的频率无明显变化。结肠癌患者MICA+单核细胞的频率与NK细胞表达NKG2D无明显关联,但与NK细胞杀伤靶细胞时CD107a的表达正相关。结论:结肠癌患者体内单核细胞表达MICA与NK细胞表达NKG2D无关,但高频率MICA阳性的单核细胞与NK细胞的抗肿瘤活性正相关。     

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.     

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.