山羊房室结的图像定量分析

目的　在于获得羊房室结组织学定量分析的详细资料。方法　采用房室结常规石蜡切片 ,HE、VanGieson、Masson染色和HPIAS 10 0 0高清晰度彩色图文分析系统进行图像分析。结果　青年羊和老年羊P细胞的面积分别是 35 2 6 μm2 、5 8 2 3μm2 ;T细胞面积分别是 4 88μm2 、136 9μm2 ;薄肯野细胞的面积分别是16 7 2 μm2 、312 2 μm2 。结论　细胞面积随年龄增长而增大 ,参数可用来区分各类细胞。     

标记滞留技术检测小鼠子宫内膜干细胞

目的：研究小鼠子宫内标记滞留细胞的存在及其分布特点。方法选择出生3天的雌性昆明白小乳鼠，实验组乳鼠皮下注射BrdU 50μg／g，对照组皮下注射生理盐水100μl，每天2次，共注射3d。于最后一次注射后2h、1w、2w、4w和8w处死并取子宫固定，石蜡包埋用于免疫组化检测，每个时间段5只乳鼠。结果实验组标记后2h，小鼠腺体和基质中标记滞留细胞（LRCs）表达最高，分别为45．1％和57．4％，随着时间的增加，LRCs逐渐降低，到第8w在腺体中仅有极少量细胞表达，而基质细胞约有1．5％的LRCs，主要位于血管周围和内膜基层连接处。不同时相腺体和基质细胞中LRCs差异具有显著性（P〈0．05）。结论昆明白小鼠子宫内LRCs主要位于血管周围和内膜基层连接处等，这些LRCs可能为子宫内膜干细胞。     

亚急性衰老小鼠脾脏T细胞CD137表达的研究

目的：探讨CD137分子在衰老小鼠脾脏T细胞表面表达的规律。方法：通过注射D－半乳糖建立亚急性衰老小鼠模型。分离衰老模型组小鼠及正常对照组小鼠的脾脏T细胞，经Con A刺激后进行培养，采用流式细胞术检查Con A刺激后不同培养时间的T细胞上CD137的表达，并进行比较。结果：小鼠脾脏T细胞经过Con A刺激后，正常对照组小鼠T细胞上CD137分子的表达高峰出现于刺激后第6天，平均表达率为48．5％，之后迅速下降。1个月和2个月衰老模型组小鼠T细胞上CD137分子在表达高峰的表达率平均分别为39．1％和32．8％，均显著低于正常对照组小鼠（P＜0．01），其中1个月模型组小鼠T细胞上CD137分子表达高峰的出现时间及下降规律与正常对照组小鼠相同；而2个月衰老模型组小鼠T细胞上CD137分子的表达高峰出现于刺激后第4天，第9天缓慢下降至与正常对照组相同时间的水平。结论：亚急性衰老小鼠T细胞上CD137分子的表达低于正常水平。至衰老过程的后期，T细胞上CD137分子的表达高峰提前出现，表达水平达到峰值后下降速度较正常小鼠缓慢，提示CD137分子在机体抗衰老过程中，具有调节T细胞功能的作用。     

人T淋巴细胞通过整联蛋白α4/VCAM-1和LFA-/ICAM-1及ICAM-2与人扁桃体生发中心结合

该文为阐明介导T细胞与淋巴细胞组织不同区域结合的粘附分子的作用，应用原位杂交粘附实验及抗体封闭试验，检测T细胞与人扁桃体组织的结合。选用针对不同抗原的单克隆抗体：CD4（13B8．2），CDlla（25．3．1），CD18（7E4）．CD28（CD28．2），CD29（K20），CD45RA（ALBll），CD49d（HP2／1），CD54（84H10），CD102（B－TI），CD106（IGll）及鼠IgM抗体；细胞选用MOLT－4，JurkatT细胞系细胞和健康人活化的T淋巴细胞。细胞培养15至20天，流式细胞仪测定表明：CD3“细胞大于98％，CD19“细胞和CD14“细胞均小于05％…     

T细胞诱导的结肠炎中多形螺旋线虫感染对CD4^＋T细胞增殖的影响

目的 研究肠道多形螺旋线虫对T细胞诱导的小鼠结肠炎CD4^±T细胞增殖情况的影响。方法 用羟基荧光素二醋酸盐琥珀酰亚胺脂（CFSE）染色的卵清蛋白（OVA）特异性CD4＾±辅助性T细胞转入重度联合免疫缺陷（SCID）小鼠中，制作小鼠实验性肠炎模型。将实验模型小鼠分为多形螺旋线虫感染组和无感染组（每组n=5），观察多形螺旋线虫感染7d后小鼠结肠炎性反应的组织学变化；以流式细胞仪检测感染3、5、7d小鼠肠系膜淋巴结中CD4＾＋T细胞CFSE的阴性率，判定肠系膜淋巴结中CD4＾＋T细胞的增殖情况。结果 与无感染组比较，感染组小鼠第7天时有螺旋线虫感染结肠炎性反应明显加重，黏膜固有层细胞浸润增多，结肠上皮破损增加，病理评分明显升高（5．20±0．84比2．00±0．71，P〈0．05）。感染3、5、7d后，感染组小鼠肠系膜淋巴结中CD4±T细胞增殖均比无感染组明显增强，CFSE的阴性率升高[3d：（7．03±1．61）％比（2．32±0．62）％，5d：（55．05±13．41）％比（29．10±2．23）％，7d：（76．97±1．89）％比（43．87±5．56）％，均P〈0．05]。结论 多形螺旋线虫感染在CD4＾＋T细胞诱导的小鼠实验性结肠炎的早期阶段促进了炎性反应的加重，可能与促进CD4＾＋T细胞的增殖有关。     

登革2型病毒43株NS1基因重组质粒DNA的免疫原性及保护作用研究

目的 研究以pcDNA3.1为载体的登革2型病毒43株（D2－43）NS1基因重组DNA的免疫原性及对登革病毒感染所致小鼠神经毒的免疫保护作用。方法 将纯化的pcDNA-NS1重组质粒DNA采用肌肉多点注射途径免疫3周龄BALB／c小鼠，剂量为每只100μg/次，检测了免疫鼠血清抗体滴度及特异性细胞毒作用。并以D2－43病毒脑内攻击6周龄BALB/c小鼠产生的神经毒症状为实验模型，对pcDNA-NS1的免疫保护作用进行了初步探讨。结果 用间接ELISA测得pcDNA-NS1免疫后抗体滴度为1：800，在补体存在下，对D2－43病毒感染的BHK－21细胞特异性杀伤率可达到61．6％。由免疫的BALB／c小鼠脾制备的效应细胞在体外可特异性地杀伤D2－43感染的P－815细胞（H－2^d)。当效靶比（E／T）为20：1时，pcDNA-NS1质粒免疫后的特异性CTL杀伤百分率为22．6％。将100 LD50的D2－43病毒经脑内攻击BALB／c小鼠，结果表明免疫pcDNA-NS1组小鼠存活率最高（90．9％）；与免疫pcDNA3.1对照组比较，P值＜0．05。结论 pcDNA-NS1质粒免疫BALB／c小鼠不仅可诱导体液免疫，还可诱导特异性细胞免疫。初步结果还显示，用含NS1基因的重组质粒DNA免疫的小鼠能免受致死剂量登革病毒的攻击，为登革热新型疫苗的研究奠定了基础。     

T-AK细胞和IL-2脂质体联合硒酸酯多糖抗白血病效应的研究

目的研究抗ＣＤ３单克隆抗体与ＩＬ－２共同诱导的Ｔ－ＡＫ细胞和ＩＬ－２脂质体（Ｌ－ＩＬ－２）联合硒酸酯多糖（ＫＳＣ）对Ｌ１２１０小鼠白血病的治疗作用。方法用ＤＢＡ／２小鼠建立Ｌ１２１０白血病模型，正常小鼠脾细胞诱生制备Ｔ－ＡＫ细胞，按设计方案转输Ｔ－ＡＫ细胞和ＩＬ－２脂质体，ＫＳＣ灌胃，检测ＮＫ细胞活性、脾淋巴细胞增殖活性和ＩＬ－２诱生水平，观察荷瘤小鼠生存期。结果Ｌ１２１０白血病小鼠的免疫功能急剧降低，生存期为１６．４３±１．９２天；转输Ｔ－ＡＫ细胞（５×１０６）和Ｌ－ＩＬ－２（１０４Ｕ／ｋｇ）能部分逆转白血病小鼠低下的细胞免疫功能，生存期延长（２４．７８±３．９４天），并有１４．３％小鼠长期存活；ＫＳＣ（４０ｍｇ／ｋｇ）对Ｔ－ＡＫ／Ｌ－ＩＬ－２的抗白血病作用有明显的增强效应，荷瘤小鼠细胞免疫功能进一步增强，生命延长率、长期存活率分别提高３６％和９９．９％。结论硒酸酯多糖具有生物反应调节剂（ＢＲＭ）样作用；以应用Ｔ－ＡＫ细胞和ＩＬ－２脂质体为主体，硒酸酯多糖为辅佐的生物治疗方案对白血病小鼠具有显著的免疫抑瘤作用     

p38及PI3K在结核杆菌抗原再刺激诱导人γδT细胞凋亡信号转导中的作用

目的：建立结核杆菌抗原（Mtb-Ag）再刺激活化的人γδT细胞凋亡模型；应用信号分子阻断剂观察p38及PI3K信号传导途径在Mtb-Ag再刺激诱导γδT细胞凋亡中的作用。方法：用3种浓度Mtb-Ag分别再刺激培养15～25天的Mtb-Ag激活的人T细胞（MtbAT）24小时后，应用Annexin-V-FITC／PI染色流式细胞术（FCM）检测γδT细胞凋亡；用Mtb-Ag（10．0μg／ml）分别再刺激MtbAT3、6、12和24小时后，观察γδT细胞凋亡情况；预先用SB203580（p38途径抑制剂）及LY294002（PI3K途径抑制剂）分别处理MtbAT60分钟，观察Mtb-Ag（10．0μg／m1）再刺激3小时后γδT细胞凋亡情况。结果：与对照组相比，10．0和20．0μg／ml MtbAg均显著诱导γδT细胞凋亡（P〈0．01），凋亡细胞比例分别增加35．63％及38．83％，且此二种浓度MtbAg在诱导γδT细胞凋亡方面无显著性差异（P〉0．05）；与对照组相比，10．0μg/ml Mtb-Ag再刺激MtbAT3、6、12和24小时均可显著诱导γδT细胞凋亡（P〈0．01），凋亡细胞比例在44．21％～51．76％之间；且Mtb-Ag再刺激MtbAT3小时实验组与再刺激MtbAT 24小时实验组相比，在诱导γδT细胞凋亡方面无显著性差异（P〉0．05），后者的凋亡细胞比例仅比前者增加7．55％；Mtb-Ag再刺激诱导γδT细胞凋亡可被SB203580（80．0μmol／L）及LY294002（10．0μmol／L）抑制，凋亡抑制率分别为91．6％及43．1％。结论：采用Mtb-Ag（10．0μg／m1）再刺激活化的γδT细胞3小时的方法，可建立Mtb-Ag再刺激活化的人γδT细胞凋亡模型；信号分子阻断剂的实验证明，p38途径及PI3K途径均参与了Mtb-Ag再刺激已活化γδT细胞凋亡过程。     

同种MHC基因胸腺转移诱导小鼠心肌移植耐受

目的：通过胸腺内注射表达外源性主要组织相容性抗原复合物（MHC）抗原质粒PXN（N2－B19－H－2Kb），诱导异基因小鼠心肌移植耐受。方法：给BALB／C小鼠胸腺内注射质粒PXN（N2－B19－H－2Kb），将外源性的编码C57BL／6小鼠MHCI类抗原的H－2KbcDNA转移到BALBC／C小鼠胸腺，2周后行C57BL／6小鼠心肌移植。用聚合酶链反应（PCR），反转录聚合酶链反应（RT－PCR），单克隆抗体免疫萤光染色流式细胞仪检测BALB／C小鼠胸腺细胞DNA，mRNA和MHC蛋白质表达。结果：BALB／C小鼠胸腺细胞表面有外源性MHC分子表达，转染效率为5．1％，胸腺内注射质粒PXN（N2－mg－H－2Kb）能明显延长移植小鼠心肌的存活时间，平均17d，对照为8d（P＜0．01）。结论：同种MHC基因转移至受体鼠胸腺可以诱导特异性的免疫耐受。     

微卫星不稳定的人类结直肠癌细胞系对5-氟尿嘧啶的敏感性

目的：探讨人类结直肠癌细胞系对化疗药物的敏感性。方法：用MTT方法测定LoVo,SW480和SW1116细胞对5－氟尿嘧啶（5－FU）的敏感性；用免疫细胞化学方法检测3细胞系hMSH2、hMLH1表达水平；用PCR－SSLP－银染法分别分析了3个细胞系基因组中10个微卫星位点状况。结果：比较3个细胞系MTT分析获得的IC50结果发现，对5－FU LoVo细胞系较SW480和SW1116更加敏感（分别为：0．8μmol/L，2.2μmol/L和1．9μmol/L，P＜0．01）。免疫细胞化学检测结果显示hMSH 2在SW480和SW1116阳性，而在LoVo阳性；hMLH1在3个细胞系均阳性。PCR－SSLP－银染法结果显示LoVo细胞系在8／10个位点和其余两个细胞系不同，电泳条带有不同程度的增加或位移，而另外的2／10个位点则具有相同的电泳带型。结论：该组微卫星位点和错配修复状况一起可以作为离体人类结直肠癌细胞系对5－FU敏感性的指标之一，为了深入阐明它们能否作为临床结直肠癌化疗病人药物筛选和预后评价的有效指标，进一步的在体研究和临床资料总结分析很有意义。     

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.