COX-2抑制剂对肺癌裸鼠移植瘤血管生成素基因表达的影响

目的：探讨环氧化酶-2(COX-2)抑制剂尼米舒利（NIM）对肺癌裸鼠移植瘤的血管生成素基因表达的影响及意义。 方法： 人肺癌A549细胞接种于裸鼠皮下，建立肺癌裸鼠移植瘤模型并予NIM治疗，计算NIM的抑瘤率，RT-PCR检测裸鼠移植瘤组织血管生成素-1、2 (Ang-1、Ang-2) mRNA表达, 免疫组织化学法测定裸鼠移植瘤组织微血管密度(MVD)。 结果： NIM可有效抑制裸鼠移植瘤的生长，其抑瘤率为43.02%。 NIM治疗组裸鼠移植瘤组织Ang-2 mRNA水平显著低于对照组（P<0.01），Ang-1 mRNA水平无显著改变（P>0.05）, Ang-2/Ang-1 mRNA比值下降（P<0.01）；同时MVD明显低于对照组（P<0.01）。 结论： COX-2抑制剂NIM可下调Ang-2基因表达,改变Ang-2/Ang-1 mRNA比值，该作用可能是COX-2抑制剂抑制肿瘤血管生成从而抑制肿瘤生长的机制之一。     

HOXB7 siRNA表达载体在裸鼠体内对人恶性黑色素瘤细胞株的影响

目的 探讨转染同源盒第7基因(HOXB7)siRNA质粒表达载体对人恶性黑色素瘤细胞株A375在裸鼠体内生长的影响.方法 裸鼠皮下接种人恶性黑色素瘤A375细胞,对2周后形成的瘤块进行分组干预.随机分为生理盐水对照组、阴性质粒组、HOXB7质粒组,观察转染后各组裸鼠移植瘤的生长情况;免疫组化法比较瘤体内微血管密度(MVD).结果 HOXB7质粒组的裸鼠移植瘤块生长慢、体积明显小于生理盐水对照组[(0.134±0.039)cm3比(1.006±0.235)cm3,P＜0.05],而阴性质粒组瘤块体积大小与生理盐水对照组无统计学差异[(0.929±0.157)cm3比(1.006±0.235)cm3,P＞0.05].HOXB7质粒组裸鼠体内肿瘤MVD低于生理盐水对照组[(2.8±1.9)比(19.9±5.6),P＜0.05],阴性质粒组与生理盐水对照组无统计学差异[(18.1±5.5)比(19.9±5.6),P＞0.05].结论 针对HOXB7 siRNA质粒表达载体可有效抑制A375细胞裸鼠体内肿瘤生长和瘤内血管生成.     

抗HER-2工程抗体chA21对人卵巢癌SKOV3裸小鼠移植瘤血管生成的影响

目的 探讨抗HER-2工程抗体chA21对过表达HER-2的人卵巢癌细胞株SKOV3裸小鼠移植瘤血管生成的影响.方法 建立人卵巢癌SKOV3裸小鼠移植瘤模型,随机分成阴性对照组和chA21组,连续5周尾静脉注射给药,观察肿瘤生长变化;5周后处死裸鼠取下瘤体,利用组织芯片及免疫组化方法结合显微图像分析系统定量检测VEGF、DLL4表达情况,并用CD31免疫组化染色比较两组微血管密度(MVD)值.结果 chA21组肿瘤组织中VEGF、DLL4的表达及MVD值均显著低于阴性对照组(P<0.05).结论 抗HER-2工程抗体chA21能显著抑制人卵巢癌SKOV3裸小鼠移植瘤的血管生成.     

结肠癌裸鼠移植瘤中Raf-1的表达与肿瘤血管生成的相关性

目的:研究Raf-1在结肠癌裸鼠移植瘤中的表达与肿瘤血管生成的关系.方法:通过Western blot方法检测不同结肠癌细胞系(HT.29、SW480、IS174T)中Raf-1的表达,然后将表达差异较大的2株细胞(HT-29、LS174T)分别接种到BALB/C nu/nu小鼠体内,分别观察肿瘤的生长速度、瘤重变化等情况,最后通过免疫组化的方法分别检测瘤组织中Raf-1的表达及肿瘤微血管密度(MVD)的情况,分析二者的相关性.结果:在裸鼠移植瘤模型中,LS174T组移植瘤生长速度及瘤重大于HT-29组;免疫组化结果显示Raf-1在LS174T组中的强阳性率为50%,在HT-29组中为10%,两组之间的差异有统计学意义(P<0.01);在LS174T组中MVD平均值为28.32±5.2,HT-29组MVD平均值为19.23±4.7,两组之间MVD值差异有统计学意义(P<0.01).结论:Raf-1是调控肿瘤组织血管生成的关键分子,在结肠癌裸鼠移植瘤中Raf-1的表达与MVD值成正相关.     

重组人血管内皮抑制素对小鼠胃癌c-Myc,bFGF表达的影响

目的 通过构建荷胃癌小鼠模型研究重组人血管内皮抑制素对癌基因c-Myc和成纤维细胞生长因子(bFGF)表达的影响及可能机制.方法 20只小鼠皮下注射MFC小鼠前胃癌细胞从而构建小鼠胃癌移植瘤模型,后随机分成两组,每组10只,观察组腹腔注射0.8mg/kg重组人血管内皮抑素,对照组腹腔注射等量生理盐水.每天观察小鼠的一般情况,每3天测量肿瘤体积,绘制肿瘤体积生长曲线并对两实验组肿瘤体积进行比较;应用RT-PCR、Western bolt及免疫组织化学法检测两组小鼠肿瘤组织中c-Myc和bFGF的表达,免疫组织化学染色法检测肿瘤组织微血管密度(MVD)的表达,并通过Spearman分析c-Myc和bFGF两者的相关性.结果 在用药的过程中,对照组小鼠随着移植瘤结节的增大而发生行为明显改变,但观察组小鼠精神、反应、活动及进食饮水均如初,未见明显不良反应.治疗后第9d开始,观察组小鼠移植瘤体积明显小于对照组(P＜0.05),在第21 d最明显(P＜0.01).RT-PCR检测显示e-Myc,bFGF mRNA相对表达量明显低于对照组(P＜0.05);Western blot检测显也显示观察组c-Myc,bFGF蛋白的表达量明显低于对照组(P＜0.05);免疫组织化学检测显示c-Myc,bFGF阳性细胞数均明显低于对照组(P＜0.05),c-Myc,bFGF平均灰度值明显高于对照组(P＜0.05);对照组平均MVD为9.2±1.3,观察组为2.9±1.0,两组之间差异亦具有统计学差异(P＜0.05).且Spearman分析得出小鼠胃移植瘤中e-Myc和bFGF mRNA存在正相关(r=0.853,P=0.000).结论 重组人血管内皮抑制素通过抑制胃癌组织c-Myc,bFGF基因表达和血管生成.     

3TSR治疗裸鼠胃癌的实验研究

目的： 探讨血管形成抑制剂3TSR对人胃癌裸鼠皮下移植瘤的抑制作用及机制。方法: 建立人胃癌裸鼠皮下种植瘤模型,实验分为对照组(腹腔注射PBS 0.2 mL)、3TSR组(腹腔注射3TSR 3 mg/kg BW)，每组8只。给药3周后处死动物,提取瘤体，测量各组荷瘤体积，HE染色检测肿瘤坏死面积百分比，免疫组织化学方法检测肿瘤微血管表达、肿瘤细胞增殖指数，原位末端标记法(TUNEL法)检测肿瘤细胞凋亡率,CD31/TUNEL/DAPI重复染色法测定肿瘤血管内皮细胞凋亡率。结果: 3TSR组平均肿瘤体积为(648.34±126.90) mm3,较对照组显著缩小(P<0.05)； 3TSR组平均肿瘤坏死面积百分比为（39.6±7.8）%,较对照组扩大69.2%，2组间有显著差异(P<0.05)。3TSR组肿瘤平均微血管数为12.8±4.1,平均微血管面积为（689.3±118.6）μm2，均显著低于对照组(P<0.05)，3TSR组增殖指数和凋亡率分别为(40.0±7.1)%和(3.4±1.2)%，与对照组比较无显著差异(P>0.05),3TSR组肿瘤血管内皮细胞凋亡率为(11.6±2.8)%,显著高于对照组的(2.9±1.5)%,P<0.05。结论: 3TSR能抑制裸鼠胃癌新生血管形成，具有显著减少肿瘤体积、肿瘤血管和增加肿瘤细胞坏死的作用，但对胃癌细胞无直接抑制作用，其机制可能与诱导内皮细胞凋亡有关。     

HOXB7 siRNA表达载体在裸鼠体内对人恶性黑色素瘤细胞株的影响

目的探讨转染同源盒第7基因(HOXB7)siRNA质粒表达载体对人恶性黑色素瘤细胞株A375在裸鼠体内生长的影响。方法裸鼠皮下接种人恶性黑色素瘤A375细胞,对2周后形成的瘤块进行分组干预。随机分为生理盐水对照组、阴性质粒组、HOXB7质粒组,观察转染后各组裸鼠移植瘤的生长情况;免疫组化法比较瘤体内微血管密度(MVD)。结果HOXB7质粒组的裸鼠移植瘤块生长慢、体积明显小于生理盐水对照组[(0.134±0.039)cm3比(1.006±0.235)cm3,P<0.05],而阴性质粒组瘤块体积大小与生理盐水对照组无统计学差异[(0.929±0.157)cm3比(1.006±0.235)cm3,P>0.05]。HOXB7质粒组裸鼠体内肿瘤MVD低于生理盐水对照组[(2.8±1.9)比(19.9±5.6),P<0.05],阴性质粒组与生理盐水对照组无统计学差异[(18.1±5.5)比(19.9±5.6),P>0.05]。结论针对HOXB7siRNA质粒表达载体可有效抑制A375细胞裸鼠体内肿瘤生长和瘤内血管生成。     

宫颈癌COX-2、MMP-9和CD105表达与肿瘤血管生成及侵袭转移的关系

目的探讨环氧合酶-2(cyclooxygenase,COX-2)、基质金属蛋白酶-9(matrix metalloproteinase-9,MMP-9)在宫颈癌局部肿瘤血管生成及肿瘤侵袭转移过程中的作用。方法采用免疫组化SP法检测32例正常宫颈上皮及68例宫颈癌中COX-2、MMP-9的表达。并用CD105标记新生血管内皮细胞,计算肿瘤内微血管密度(microvascular density,MVD)。结果COX-2、MMP-9在宫颈癌组织中的阳性表达率均显著高于正常宫颈上皮中的表达(P<0.01)。COX-2、MMP-9阳性组的MVD均显著高于COX-2、MMP-9阴性组(P<0.01),淋巴结转移组COX-2、MMP-9阳性率均高于无淋巴结转移组(P<0.05)。结论COX-2、MMP-9阳性表达可能在宫颈癌血管生成和侵袭转移中起重要作用,两者在宫颈癌血管生成中可能具有协同作用。检测宫颈癌中COX-2、MMP-9表达对进一步了解宫颈癌局部血管生成情况及判断预后有一定的应用价值。     

黄芪、党参提取物增强紫杉醇抑制肿瘤血管生成和转移的实验研究

目的:观察黄芪、党参提取物对紫杉醇抑制Lewis肺癌血管生成和肿瘤转移的增强作用,为中医药抗肿瘤应用提供实验依据.方法:C57BL/6小鼠右肋处皮下接种Lewis肺癌,6 h后随机分为实验组、紫杉醇组和对照组.实验组接受紫杉醇联合参芪注射液治疗,紫杉醇组接受同等剂量的紫杉醇治疗,对照组用等体积的生理盐水代替.观察指标:移植瘤体积,肿瘤组织内微血管度和肺脏转移瘤数量,小鼠存活时间.结果:与紫杉醇治疗相比,紫杉醇联合参芪可以明显减少移植瘤内的微血管密度(10.1±4.4 vs 16.8±7.3,P＜0.05)和肺脏转移瘤个数(13.4±4.3 vs 18.4±3.9,P＜0.05),明显延长小鼠的存活时间(43.3±8.4 d vs 36.3±6.6 d,P＜0.05). 结论:黄芪、党参提取物对紫杉醇抑制肿瘤血管生成和移植肿瘤转移有一定的增强作用.     

NK4基因转染对裸鼠人淋巴瘤移植瘤的抑制作用

目的 探讨NK4基因转染对裸鼠人淋巴瘤移植瘤的抑制作用及其机制.方法 采用NK4基因重组质粒pVITRO2-NK4转染的Raji细胞建立裸鼠皮下人淋巴瘤移植瘤模型,动态监测裸鼠体重和肿瘤大小.8周后获取瘤组织,分别采用免疫组化和脱氧核糖核酸末端转移酶介导的缺口末端标记法(TdT-mediated dUTP nick end labeling,TUNEL)检测移植瘤组织的细胞凋亡和微血管密度(microvessel density,MVD),并进行相关分析.结果 NK4基因转染组裸鼠移植瘤体积明显小于对照组(P<0.01),而对照组间差异无显著性(P>0.05);各组间裸鼠体重降低,差异无显著性(P<0.01).NK4基因转染组的淋巴瘤细胞AI值达237±10.94,而质粒pVITRO2转染组和未转染Raji组的AI值分别为79.7±30.8和81.7±22.2,NK4基因转染组明显高于对照组(P<0.001),而对照组间差异无显著性(P>0.05);NK4基因转染组MVD为4.7±1.52,而质粒pVITRO2转染组和未转染Raji组MVD分别为12.0±1.00和10.66±1.53,NK4基因转染组明显低于对照组(P<0.001),而对照组间差异无显著性(P>0.05).结论 NK4基因转染可明显抑制裸鼠人淋巴瘤移植瘤的生长,其可能通过抑制肿瘤血管新生和促肿瘤细胞凋亡而发挥其效应.     

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.     

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 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.