乳腺癌组织中ERα mRNA和PR mRNA检测方法的建立和应用

建立测定雌激素受体α(ERα)mRNA和孕激素受体(PR)mRNA的实时荧光定量RT-PCR,探讨两者在乳腺癌组织及良性乳腺肿瘤组织中的表达水平。分别以pMD18 ERα和pMD18-PR质粒为定量模板,用循环阈值(Ct)定量起始模板,在荧光TaqMan方法的基础上建立了测定ERαmRNA和PR mRNA的实时荧光定量RT-PCR,并分别测定48例乳腺癌组织及28例良性乳腺肿瘤组织中ERαmRNA和PR mRNA的表达水平。ERαmRNA和PR mRNA测定的线性范围为10~3～10~8copy/μg RNA;ERαmRNA和PR mRNA高值和低值的批内和批间变异系数(CV)在5.07%～11.28%之间。ER mRNA在乳腺癌组织中的表达量为6.76×10~5copy/μg RNA(4.17×10~5,9.34×10~5),良性乳腺肿瘤组织中的含量为1.54×10~5 copy/μg RNA(1.02×10~5,2.06×10~5),乳腺癌组织的表达量高于良性组织(P<0.05);PR mRNA在乳腺癌组织中的表达量为1.02×10~6copy/μg RNA(6.81×10~5,1.36×10~6),良性乳腺肿瘤组织中的含量为4.93×10~5 copy/μg RNA(3.21×10~5,6.65×10~5),两者表达无明显差异(P>0.05)。ERαmRNA和PR mRNA在乳腺癌和良性乳腺肿瘤组织中的表达存在相关性。我们建立的测定ERαmRNA和PR mRNA的实时荧光定量RT PCR灵敏、稳定、重复性好,可供临床检测和研究。ERαmRNA和PR mRNA基因表达水平可作为预测治疗效果及判断预后的重要指标。     

左胫骨近端肌纤维母细胞瘤（Myofibroblastoma）

患者,女性,43岁左小腿痛性肿物6月余,半年前无明显诱因出现左胫骨上段疼痛,可触及1小包块,红肿压痛,生长缓慢、逐渐增大.2个月前局部肿胀疼痛加剧.10年前因输卵管结核曾作手术.血RBC4·07×1012/L;WBC5·5×109/L;ESR23mm/h;血碱性磷酸酶:47μ/L谷草转氨酶:20μ/L,谷丙转氨酶12μ/L;尿本周蛋白阴性.手术见:左胫骨23cm一段,切面骨髓腔内10cm×3cm×3cm肿物,灰白色,带液,边界清楚,部分呈鱼肉状,侵犯骨皮质.进行左胫骨肿瘤段切除,异体骨移植,AO钢板内固定,人工骨关节表面置换、皮瓣转移术,术后化疗.骨炎症性肌纤维母细胞瘤影像学图片说…     

顺铂的多靶性对癌细胞成分的作用

用昆明种小鼠腹水肝癌细胞3×10~6/ml,给顺铂10μg/ml,分别作用1、2、5、14、24小时。给药组和对照组分别用Rhodamine-phalloidin 10μg/ml标记癌细胞膜下和分离的核内肌动蛋白微丝,吖啶橙10 μg/ml标记分离的核内DNA,FITC 10μg/ml标记分离的核内染色质蛋白,用MPV-2显微分光光度计进行观察和测定含量变化。顺铂作用1小时后,癌细胞膜下肌动蛋白微丝     

钴60放射法制备免疫功能抑制大鼠模型

目的通过系统的钴60放射计划,对大鼠免疫功能抑制的合适剂量进行评估,为干细胞移植研究提供合适的的动物模型。方法SD大鼠随机分为12.5Gy、10Gy、7.5Gy、5Gy4个剂量组,10只/组。计算放射后2个月的死亡率。检测白细胞数量,评价各剂量组动物的白细胞参考值范围。结合死亡率和白细胞参考值范围,确定合适的放射剂量。通过对外周血中性粒细胞(PMN)吞噬实验和外周血白细胞移行抑制实验(MIT),对细胞免疫功能进行检测。结果行12.5Gy照射的大鼠,3d内全部死亡;10Gy放射后的大鼠在1个月内全部死亡;7.5Gy照射的大鼠2个月内死亡1只;5Gy照射的大鼠没有死亡情况发生。设定95%作为可信区间,计算白细胞数量参考值范围(109个/L),未放疗组:16.978+1.96×6.46;5Gy放疗组:4.93+1.96×0.72;7.5Gy放疗组:2.313+1.96×0.782;10Gy放疗组:1.03+1.96×0.507。t检验表明,随着放疗剂量的增加,各实验组的白细胞数量显著减少。对于细胞免疫功能的检测,外周血中性粒细胞(PMN)吞噬实验结果表明,机体全身免疫功能降低;外周血白细胞移行抑制实验(MIT)表明,机体T细胞免疫功能降低。结论7.5Gy可以一定程度上抑制大鼠的免疫功能,为合适的放疗剂量。     

左胫骨近端肌纤维母细胞瘤（Myofibroblastoma）

患者,女性,43岁左小腿痛性肿物6月余,半年前无明显诱因出现左胫骨上段疼痛,可触及1小包块,红肿压痛,生长缓慢、逐渐增大.2个月前局部肿胀疼痛加剧.10年前因输卵管结核曾作手术.血RBC 4.07×1012/L;WBC 5.5 ×109/L;ESR 23mm/h;血碱性磷酸酶:47μ/L谷草转氨酶:20μ/L,谷丙转氨酶12μ/L;尿本周蛋白阴性. 手术见:左胫骨23cm一段.切面骨髓腔内10cm×3cm×3cm肿物,灰白色,带液,边界清楚,部分呈鱼肉状,侵犯骨皮质.进行左胫骨肿瘤段切除,异体骨移植,AO钢板内固定,人工骨关节表面置换、皮瓣转移术,术后化疗.     

重组人内抑素对人瘢痕疙瘩成纤维细胞形态及增殖的影响

目的 观察重组人内抑素(rhEndostatin)对体外培养的人瘢痕疙瘩成纤维细胞(KFs)增殖的影响。方法 体外培养及鉴定人KFs; 应用四甲基偶氮唑盐（MTT）法分别检测不同浓度rhEndostatin (6.25×10^-6、1.25×10^-5、2.50×10^-5、5.00×10^-5、1.00×10^-4mg/L)作用24h、48h、72h后对KFs增殖的影响,并同时观察其形态变化。结果 组织块接种7~8d后,其周边可见少许梭形细胞,继之细胞逐渐增多并以组织块为中心呈放射状生长;传2代培养的细胞单层排列,形态均一,呈长梭状,折光性好。细胞波形蛋白免疫化学染色显示胞质均呈棕黄色。rhEndostatin(6.25×10^-6 mg/L浓度组除外)能明显抑制KFs的增殖(P <0.05),抑制效应与rhEndostatin浓度和作用时间呈一定的依赖关系; 其中,5.00×10-⁵ mg/L 和1.00×10^-4 mg/L组rhEndostatin作用48 h后,细胞生长缓慢,体积变小,数目减少,排列紊乱。结论 重组人内抑素对人KFs增殖具有抑制作用。     

免疫小鼠脾脏RNA引起的免疫记忆参与细胞

本工作用体外抗体产生系统,研究了由免疫核糖核酸(iRNA)引起的、参与免疫记忆的T细胞与B细胞.实验动物采用8-12周龄的BALB/C小鼠及由BALB/C小鼠培育的裸鼠.抗原用10%绵羊红细胞(SRBC)或马红细胞(HRBC).取0.1ml抗原给小鼠作腹腔内注射,四周后再加强注射.免疫后五天将小鼠解剖,用改进Kidson法自小鼠脾脏提取iRNA.取500μgiRNA给小鼠作腹腔注射,四周后再加强注射.脾细胞的制备及试管内抗体形成的测定:在双玻皿内将小鼠脾脏切开,加入培养液制备成无菌的单个细胞悬液.在1ml RPMl1640培养液内计数脾细胞1×10~7,并加入青霉素G50U/ml、硫酸链霉素50μg/ml、硫烷醇(5×10~(-5)M),再补足20%胎     

IL-2对于鼠流感病毒感染的预防效果

IL-2具有促进杀伤T 细胞增殖分化,诱导LAK 细胞等多种生物学作用。这些作用主要是提高抗体的产生、增强细胞免疫功能。据此推测IL-2在防止感染方面也起重要作用。本文通过给与小鼠IL-2,观察其预防流感病毒感染的效果。实验动物为9周龄雌性ICR 系小鼠。流感病毒是采用鸡受精卵增殖的A/PR/8/34(HINI)病毒,经鼻接种5LD50病毒给小鼠。每只鼠于腹腔内分别给予1mlIL-2,含量分别为5×10~4、1×10~4、5×10~3、1×10~3国际单位(IU)。对照组给予等量     

ATG及ALG治疗2例重症再生障碍贫血的疗效

本文报道用ATG(抗胸腺细胞球蛋白)和ALG(抗淋巴细胞球蛋白)治疗2例重症再生障碍贫血,效果显著。例1,男性23岁,入院时血液检查:红细胞171×10~4/μl、血红蛋白(Hb)6.5g/dL、血小板2.1×10~4/μL,各类血细胞严重减少。应用大量强的松龙(MPSL)治疗2个疗程无效。给予ATG(20mg/kg/日)静点,总量5g,10周后取得良好效果,骨髓穿刺检查有核细胞由1.5×10~4/μL 堵至11.5×10~4/μL。一年后Hb 增至15.7g/dL、颗粒细胞1580/μL、血小板9.2×10~4/μL,ATG 的治疗效果显著。     

人生长激素的生物素—新和素放大酶联免疫法的建立

为建立有生物素和亲和素放大系统的人血清生长激素 (hGH)酶联免疫分析法 (BA ELISA) ,以微量的GH抗原免疫BALB/C小鼠 ,利用杂交瘤技术制备出 2 9株分泌抗hGH单克隆抗体 (MCA)的杂交瘤株。选择最佳配对MCA ,利用亲和素与生物素高亲和力的特性 ,建立了血清hGH双位点的BA ELISA法。我们的MCA滴度为 (0 5～ 5 0 0 )× 10 4,亲和常数Ka =(0 13～ 1 40 )× 10 10 L/mol。血清hGH的ELISA的灵敏度为 0 0 4± 0 0 1μg/L ;在血清中分别加入 1,2 5 ,5 μg/L的hGH ,其回收率为 90 7%～ 10 7 6 % (平均为 10 1 2 0 %± 0 0 3 % ) ;hGH含量为 2 8、10 0、2 9 8μg/L的血清批内、批间变异系数分别为 4 9%、3 8%、7 9%和 6 2 %、3 5 %、9 5 % ;用本法测定了正常儿童、生长激素缺乏症 (GHD)患儿和活动性肢端肥大症患者空腹血清hGH水平 ,分别为 1 87± 3 0、0 30± 0 42和 8 71± 6 95 μg/L。上述结果表明 ,本ELISA方法灵敏、特异、稳定、准确 ,能确切反映hGH分泌功能 ,在诸多方面均优于RIA。     

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.     

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.     

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.     

The universal muscular chain reaction,muscle spasm,Torsions, ruptures and extravasations. Chameleons of pathology and some manifestations of simple muscular disorders

Most bodily functions require the coordinated actions of complementary and supplementary paired muscle groups. Where this essential muscular cooperation is lacking, hollow organs may burst and others become literally screwed up, giving rise to many similar spastic diseases such as Torticollis, Twisted ovarian cyst, Torsion of the Testis, Volvulus of the intestines, Varicose Veins, Megacolon, Aortamegaly, Scoliosis, Erb's Palsy, Peyronie's Disease, Main-en-Griffe, Undescended Foot (Pes Cavus), Talipes, Strabismus. Spasm is “panenepidemic” and unclassified examples of Torsion Dystonia and Dyskinesia really are as common as debt and taxes.     

Class II HLA in Georgia Caucasus Tbilisi Georgians and their Mediterranean ancestry: The Usko Mediterranean languages

Georgia (or Sakartvelo in its own language) is a South Caucasus Mts. country with its easternmost part is enigmatically named Iberia, like the Iberian Peninsula, which may refer to rivers “Kura” and “Ebro” or their valleys respectively. Most of their inhabitants speak Georgian which is included within Dene-Caucasian group and Usko-Mediterranean subgroup of languages. The latter includes Basque, Berber, ancient Iberian-Tartessian, Etruscan, Hittite, Minoan Lineal A and others. In the present paper, HLA class II -DRB1 and -DQB1 alleles has been studied and extended haplotypes calculated. Most frequent haplotypes are also of Mediterranean origin (i. e.: (A*02-B*51)-DRB1*11:01-DQB1*03:01, (A*02-B*51)-DRB1*13:01-DQB1*06:03, or (A*24-B*35)-DRB1*01:01-DQB1*05:01) and DA genetic distances show that closest world populations to Georgians are Mediterraneans. Georgians also show common extended haplotypes ((A*02-B*51)-DRB1*11:01-DQB1*03:01, (A*02-B*13)-DRB1*07:01-DQB1*02:01 and (A*03-B*35)-DRB1*11:01-DQB1*03:01) with Svan people, a secluded population in North Georgia mountains. We can conclude that Georgians belong to a very old Mediterranean substratum according to both linguistics (Usko Mediterranean languages) and HLA genetics.     

'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.