慢性活动性肝炎的胆小管增生机制——人肝多种胶原间质免疫组化研究

作Carnoy固定的肝穿刺活检50例(前)Ⅰ、Ⅲ、Ⅳ、Ⅴ、Ⅳ型胶原及FN、LM的免疫组化研究。用ABC法,其中慢活肝17例。结果首次发现人类胆管上皮Ⅴ型胶原强阳性(其它型别阴性);由此证实了人肝小叶内有“毛细管样胆小管”存在;后者在慢活肝时大量增生,并穿插肝索。认为这可能是慢活肝时纤维化的组织机制。     

人参皂苷Rd干预体外培养肝储脂细胞胶原的表达

背景：肝储脂细胞产生胶原是导致肝硬化的直接原因,调控肝储脂细胞产生胶原的能力即可以防治肝硬化。 目的：观察人参皂苷Rd对体外培养肝储脂细胞Ⅰ,Ⅲ型胶原表达的影响。 方法：体外培养肝储脂细胞,随机分为对照组和实验组,实验组用100 mg/L人参皂苷Rd进行培养干扰,用免疫组织化学、免疫荧光技术检测Ⅰ,Ⅲ型胶原基因的表达。 结果与结论：实验组比对照组,Ⅰ,Ⅲ型胶原阳性产物吸光度和面积比值明显降低(P < 0.05),说明人参皂苷Rd对体外培养肝储脂细胞Ⅰ,Ⅲ型胶原的表达有明显调节作用,影响肝储脂细胞产生胶原纤维。     

CCl_4诱导大鼠肝纤维化动物模型Ⅰ、Ⅲ及Ⅳ型胶原生成变化的研究

目的探讨肝纤维化过程中胶原生成细胞的来源以及Ⅰ、Ⅲ、Ⅳ型前胶原基因及其蛋白表达的演变规律。方法利用免疫组化及核酸分子杂交技术，对ＣＣｌ４诱导ＳＤ大鼠肝纤维化不同阶段（２０周内）α１（Ⅰ）、α１（Ⅲ）及α１（Ⅳ）前胶原ｍＲＮＡ的表达进行动态观察。结果（１）肝纤维化时α１（Ⅳ）前胶原ｍＲＮＡ早期即迅速升高，α１（Ⅲ）前胶原ｍＲＮＡ呈优势性表达，α１（Ｉ）前胶原ｍＲＮＡ含量的增加则较为缓慢；（２）肝纤维化早期，变性坏死区结蛋白阳性和α－平滑肌肌动蛋白阳性的Ｉｔｏ细胞及肌纤维母细胞表达α１（Ⅲ）、α１（Ⅳ）及α１（Ⅰ）前胶原ｍＲＮＡ。纤维化中、后期，三种前胶原ｍＲＮＡ表达主要见于间隔内纤维母细胞和肌纤维母细胞。结论二者构成肝纤维化时的主要胶原生成细胞。此外，窦内皮细胞也参与肝内Ⅳ型胶原的合成。     

肝病患者血清Ⅲ型胶原抗体的检测分析

肝病患者血清Ⅲ型胶原抗体的检测分析陈黎明李跃旗辛绍杰程云张士兰张晓峰赵军董漪张鸿飞肝纤维化是肝硬变的早期，对肝纤维化的全面了解，将有助于肝硬变防治或逆转〔１〕。肝纤维化发生时，Ⅲ型胶原和其它胶原在肝脏内大量沉积，同时血清中也有一定量的胶原游离，与肝脏...     

抗IGFBPrP1抗体对小鼠肝纤维化的预防作用及其机制的研究

目的: 明确抗胰岛素样生长因子结合蛋白相关蛋白1(IGFBPrP1)抗体能否预防硫代乙酰胺(TAA)诱导的小鼠肝纤维化的形成,同时探讨其机制。方法: 将24只雄性C57BL/6野生型小鼠随机分为正常对照组、TAA 4周组和TAA+抗IGFBPrP1抗体4周组,每组8只,观察肝组织形态学改变,免疫组织化学染色和Western blotting检测肝组织中α-平滑肌肌动蛋白(α-SMA)、转化生长因子β₁(TGF-β₁)、Smad3、磷酸化Smad2/3(p-Smad2/3)、纤维连接蛋白(FN)、Ⅰ、Ⅲ型胶原(collagen Ⅰ、Ⅲ)及IGFBPrP1的表达。结果: TAA 4周组肝损伤严重,α-SMA、TGF-β₁、Smad3、p-Smad2/3、FN、collagen Ⅰ、Ⅲ及IGFBPrP1的表达明显高于正常对照组(P＜0.01),TAA+抗IGFBPrP1抗体4周组肝损伤减轻,上述各指标表达均低于TAA 4周组(P＜0.01)。IGFBPrP1与TGF-β₁、Smad3、p-Smad2/3 、FN及collagen Ⅰ的表达呈正相关(P＜0.01)。结论: 抗IGFBPrP1抗体可预防TAA诱导的小鼠肝纤维化的形成,其机制为抑制肝星状细胞的活化和减少细胞核内p-Smad2/3的表达、抑制TGF-β₁/Smad3信号通路,进而导致细胞外基质在肝组织中沉积减少。     

Ⅲ型胶原肾小球病的形态学观察

目的了解Ⅲ型胶原肾小球病的形态学改变,并对Ⅲ型胶原可能的细胞来源进行初步探讨。方法对3例肾活检组织进行光镜、免疫荧光、电镜和Ⅰ、Ⅲ、Ⅳ型胶原及d平滑肌肌动蛋白(α-SMA)的免疫组织化学染色(SP法)观察。结果2例患者临床表现为肾病综合征,其中1例伴高血压,第3例表现为肾功能不全和肾性高血压。3例均无肾病家族史。光镜检查可见肾小球基膜内和系膜区弥漫性过碘酸-希夫反应阳性物质沉积,系膜细胞无明显增生。电镜检查在基膜内疏松层和系膜区可见大量胶原纤维沉积,系膜细胞胞膜下平行排列的束状微丝明显增加。免疫组织化学显示这些胶原纤维为Ⅲ型胶原,Ⅰ型和Ⅳ型胶原阴性,同时系膜区多数系膜细胞α-SMA阳性。结论Ⅲ型胶原肾小球病光镜、电镜及免疫组织化学上都有其特殊的病理改变。肾小球内激活的系膜细胞可能是Ⅲ型胶原的来源。     

神经生长因子对肝星状细胞胶原分泌及形态学的影响

目的: 观察神经生长因子(NGF)对大鼠肝星状细胞(HSCs)胶原分泌功能和形态学的影响,探讨NGF抗肝纤维化的作用机制。方法: 大鼠肝星状细胞株(HSCs-T6)经不同浓度NGF作用24 h后,ELISA法检测上清液中Ⅰ、Ⅲ型胶原的含量;通过倒置显微镜、吖啶橙染色及透射电镜观察形态学变化。结果: NGF作用HSCs 24 h后培养上清中Ⅰ、Ⅲ型胶原的含量与对照组相比明显减少(P<0.05),但无剂量依赖性。倒置显微镜下可见细胞数目变少,细胞变小,形态向椭圆形或圆形改变;吖啶橙染色及透射电镜可以见到细胞凋亡的形态学变化。结论: NGF可抑制HSCs合成Ⅰ、Ⅲ型胶原,诱导HSCs凋亡,抑制HSCs胶原分泌,可能是NGF抗肝纤维化的作用机制。     

CCl诱导大鼠肝纤维化动物模型Ⅰ,Ⅲ及Ⅳ型胶原生成变化的研究

探讨肝纤维化过程中胶原生成细胞的来源以及Ⅰ、Ⅲ、Ⅳ型前胶原基因及其蛋白表达的演变规律。     

大鼠早期肝纤维化α_1(Ⅰ)、α_1(Ⅲ)前胶原mRNA及相应胶原蛋白表达的初步研究

应用Northern杂交及免疫组化方法.观察CC1_4诱导的大鼠肝纤维化形成不同阶段α_1(Ⅰ)、α_1(Ⅲ)前胶原mRNA表达与相应Ⅰ、Ⅲ型胶原蛋白ColⅠ、ColⅢ及前Ⅲ型胶原端肽(PⅢP)表达间的变化规律及其相互关系。结果表明:①肝纤维化时α_1(Ⅰ)、α_1(Ⅲ)前胶原mRNA表达增强,早期以α_1(Ⅲ)mRNA为主;α_1(Ⅰ)及α_1(Ⅲ)前胶原mRNA均于3～4周时呈一过性下降。②Ito细胞是肝纤维化过程中产生Col Ⅰ、ColⅢ及PⅢP的主要来源,肝细胞亦具有一定的合成ColⅢ及PⅢP的能力。③应用Northern杂交技术检测前胶原mRNA表达,能从基因水平了解肝内胶原含量的变化,能更敏感地反映肝纤维化的倾向。     

成纤维细胞饲养层饲养人角朊细胞的机制研究

目的 探讨人成纤维细胞(HFB)是否可作为饲养层支持人角朊细胞(KC)的生长.方法 分离培养人真皮FB,用丝裂霉素C处理制成饲养层,应用RT-PCR、免疫细胞化学染色方法检测Ⅰ型、Ⅲ型前胶原mRNA和胶原蛋白的表达;在FB饲养层上接种人KC,观察其促进KC贴壁、生长、移行及分化的作用;并设置NIH3T3细胞饲养层为对照组,在其上接种人KC,观察KC贴壁数和膜片完全融合时间.结果 FB饲养层Ⅰ型前胶原mRNA的表达有所增加(P＜0.05)、Ⅲ型前胶原mRNA的表达无明显变化,抗Ⅰ型、Ⅲ型胶原蛋白抗体阳性;以适宜密度接种的成纤维细胞饲养层,KC贴壁、生长和分化良好;二种饲养层KC贴壁数和膜片完全融合时间无差异.结论 人真皮FB饲养层能够分泌Ⅰ型和Ⅲ型胶原,促进人KC生长并形成复层细胞.     

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.