激活素A对RAW264.7巨噬细胞活性的调节作用

目的探讨激活素A对参与炎症反应的小鼠巨噬细胞活性调节作用。方法以LPS刺激活化的小鼠巨噬细胞系RAW264．7细胞作为阳性参照,ELISA法检测激活素A及LPS刺激的小鼠腹腔巨噬细胞系RAW264．7细胞IL-1β分泌水平,还原酶法分析NO分泌水平,RT-PCR检测IL-1β和iNOS mRNA的表达,瑞氏染色检测RAW264．7细胞吞噬活性。结果在激活素A刺激下RAW264．7细胞IL-1β和NO分泌水平均明显升高,IL-1β和iNOS mRNA表达亦增加,巨噬细胞吞噬活性增强;激活素A和LPS共刺激RAW264．7细胞时,激活素A明显抑制LPS刺激的RAW264．7细胞IL-1β和NO产生水平,以及IL-1β和iNOS mRNA表达,巨噬细胞吞噬活性也明显低于LPS单独刺激组。结论激活素对巨噬细胞的活性调节具有双重作用,这种作用与巨噬细胞的激活状态有关。     

需钙蛋白酶抑制剂Ⅰ对LPS攻击的RAW264．7细胞中iκBα表达和细胞因子分泌的影响

目的：探讨需钙蛋白酶抑制剂I（CI-I）对LPS攻击RAW264．7细胞后iκBα表达和细胞因子分泌的影响。方法：用CI—I预处理RAW264．7细胞1h后，再用LPS攻击，分别用Western blot和ELISA检测RAW264．7细胞iκBα蛋白表达和TNF-α、IL-6的分泌。结果：CI—I预处理RAW264．7细胞后，可抑制LPS导致的iκBα表达降低。LPS攻击RAW264．7细胞后4h和8h，TNF—α和IL-6的分泌均增加，CI—I和地塞米松（DEX）可抑制该效应，并具有协例抑制作用。结论：CI—I和DEX可抑制LPS攻击后RAW264．7细胞中iκBα表达和炎性细胞因子分泌，从而有助于减轻细胞损伤。     

GW4064 对LPS 诱导的RAW264.7 细胞中炎症反应的影响

目的:研究FXR激动剂GW4064对脂多糖(LPS)诱导的RAW264.7巨噬细胞炎症因子TNF-α、MCP-1和NO的分泌及IL-1β、TNF-α的表达,以及诱导型一氧化氮合酶(iNOS)表达的影响。方法:通过流式高通量多因子检测技术和Griess法检测炎症因子TNF-α、MCP-1和NO分泌的影响;采用qPCR技术检测炎症因子IL-1β、TNF-α和iNOS的mRNA的表达;Western blot检测iNOS及NF-κB蛋白的表达的水平。结果:GW4064能抑制LPS诱导的细胞促炎因子IL-1β、TNF-α和iNOS的mRNA的表达(P0.05),同时能不同程度的抑制细胞上清中TNF-α,MCP-1中的分泌量(P0.05);并且呈剂量依赖性的抑制NO的释放(P0.05);除此之外,GW4064(10、20μmol/L)可显著抑制LPS诱导的RAW264.7巨噬细胞iNOS蛋白的产生(P0.01),经过GW4064处理后,LPS诱导的RAW264.7巨噬细胞NF-κB的活性也显著下调(P0.01)。结论:FXR激动剂GW4064在巨噬细胞中具有一定的抗炎作用,抑制iNOS的表达及下调NF-κB的活性是其发挥抗炎作用的机制之一。     

双氢青蒿素对CpG ODN诱导小鼠RAW264.7细胞释放细胞因子的影响

目的：观察双氢青蒿素对CpG ODN诱导小鼠RAW264．7细胞释放细胞因子的影响。方法：体外培养小鼠RAW264．7细胞．用CpG ODN刺激小鼠RAW264．7细胞分泌TNF-α、IL-6,用不同浓度双氢青蒿素进行拮抗,ELISA法检测培养上清中促炎细胞因子的含量．结果：双氢青蒿素可抑制CpG ODN刺激小鼠RAW264．7细胞释放TNF—α和IL-6,并呈一定的量效关系,其中对IL-6最大抑制率可达到66．5％,而对TNF—α释放的最大抑制率仅为7．2％,其影响细胞因子释放的作用与细胞毒作用无关。结论：双氢青篙素呈浓度依赖性地抑制CpG ODN刺激小鼠RAW264．7细胞释放TNF—α和IL-6,但对TNF—α的影响较小。     

没食子酸通过拮抗脂多糖诱导的TLR4/NF-κB活化抑制RAW264.7巨噬细胞炎性反应

目的观察没食子酸对脂多糖(LPS)诱导RAW264.7巨噬细胞Toll样受体4/核因子κB(TLR4/NF-κB)通路的影响。方法将巨噬细胞分为空白对照组、LPS组、LPS联合没食子酸组、LPS联合NF-κB抑制剂吡咯二硫代甲酸(PDTC)组和LPS联合地塞米松(DM)组。处理后的细胞培养24 h,ELISA检测肿瘤坏死因子α(TNF-α)、白细胞介素1(IL-1)、IL-6水平,实时定量PCR检测TLR4、NF-κB mNRA水平,Western blot法检测p65、p-p65、TLR4、磷酸化的NF-κB抑制蛋白α(IκBα)的蛋白表达水平。结果 LPS诱导RAW264.7巨噬细胞后TNF-α、IL-1、IL-6水平升高,没食子酸可降低LPS诱导引起的TNF-α、IL-1和IL-6表达水平升高。LPS刺激后TLR4 mRNA及蛋白表达增加,NF-κB活化,没食子酸可拮抗以上作用,阻止NF-κB活化。结论没食子酸可通过TLR4/NF-κB通路抑制LPS诱导的RAW264.7巨噬细胞炎性反应。     

川续断皂苷Ⅵ对M1/M2型巨噬细胞极化的调节作用及机制

目的：研究川续断皂苷Ⅵ对M1/M2型巨噬细胞极化的调节作用及机制。方法：MTT法检测川续断皂苷Ⅵ对RAW264.7细胞活力的影响。ELISA检测脂多糖（LPS）诱导状态下RAW264.7细胞上清中TNF-α和IL-6的分泌量；Griess法检测LPS诱导状态下RAW264.7细胞上清中一氧化氮（NO）含量；荧光定量PCR检测TNF-α、IL-6、精氨酸酶-1(Arg-1)、血红素加氧酶1(HO-1)和细胞因子信号转导抑制蛋白-2(SOCS2)基因表达水平。Western blot检测一氧化氮合酶（iNOS）和p-p65蛋白表达。结果：在LPS诱导的RAW264.7细胞中，川续断皂苷Ⅵ抑制TNF-α、IL-6、iNOS和p-p65蛋白或基因表达水平，同时增加HO-1基因表达。川续断皂苷Ⅵ能抑制LPS诱导下RAW264.7细胞分泌的NO。川续断皂苷Ⅵ增加IL-4诱导下M2型巨噬细胞标志物Arg1和SOCS2基因表达。结论：川续断皂苷Ⅵ可以抑制RAW264.7巨噬细胞向M1型极化，同时促进其向M2型极化，可通过调节M1/M2型巨噬细胞极化发挥其抗炎免疫调节作用。     

瘤果黑种草子总皂苷的抗炎和免疫调节作用

目的 考察瘤果黑种草子总皂苷(TSSN)的抗炎和免疫调节作用。方法 建立脂多糖(LPS)诱导的小鼠单核巨噬细胞白血病细胞RAW264.7炎症模型，采用Griess法检测NO分泌水平，ELISA法检测细胞上清TNF-α、IL-6、IL-1β含量，采用qRT-PCR方法考察TSSN对NF-κB信号通路相关靶点mRNA表达水平的影响；酶动力学实验检测总皂苷对环氧酶-2(COX-2)以及5-脂氧酶(5-LO)的抑制活性；通过MTS法及ELISA法检测TSSN对静息期淋巴细胞的影响，并以TSSN对刀豆蛋白(ConA)、LPS分别诱导的小鼠T、B淋巴细胞增殖的影响，以及对T淋巴细胞分泌细胞因子TNF-α、IL-2、IL-6、IFN-γ的影响来考察TSSN的免疫调节作用。结果 在LPS诱导的RAW 264.7炎症模型中，TSSN能显著降低LPS诱导的RAW264.7分泌炎症介质NO和促炎症因子TNF-α、IL-6、IL-1β水平，显著抑制TNF-α、IL-6、IL-1β m RNA表达水平，抑制iNOS、NF-κB p65 mRNA表达水平；在12.5～200μg·ml^-1剂量...     

HBV X蛋白即时高表达上调巨噬细胞分泌TNF-α和IL-1β

目的：将乙型肝炎病毒（HBV）X蛋白真核表达载体pcDNA3.1（＋）-HBx转染巨噬细胞,观察X蛋白即时高表达对巨噬细胞分泌细胞因子TNF-α、IL-1β的影响,为进一步研究HBVX蛋白的致病机制奠定实验基础。方法：将真核表达载体pcDNA3.1（＋）-HBx通过SuperFectTM脂转染试剂转染巨噬细胞24小时后,用LPS刺激巨噬细胞,利用Westernblot鉴定X蛋白在巨噬细胞中的表达,并用酶联免疫吸附试验（ELISA）检测pcDNA3.1（＋）-HBx转染的巨噬细胞不同时间（12、24、48小时）培养上清液中细胞因子TNF-α、IL-1β的含量,统计软件SPSS13.0分析所得数据。结果：将真核表达载体pcDNA3.1（＋）-HBx转染巨噬细胞,Westernblot结果显示pcDNA3.1（＋）-HBx能在巨噬细胞中表达约17kD的目的蛋白,即X蛋白;ELISA法测定细胞上清液中TNF-α、IL-1β的含量,LPS刺激组及pcDNA3.1（＋）组与空白对照组有显著性差异（P〈0.01）,pcDNA3.1（＋）-HBx组与LPS刺激组及pcDNA3.1（＋）组有显著性差异（P〈0.01）,而LPS刺激组与pcDNA3.1（＋）组无显著性差异（P〉0.05）,即LPS可活化巨噬细胞分泌细胞因子,且X蛋白即时高表达可引起LPS诱导的巨噬细胞分泌TNF-α和IL-1β明显增加。结论：乙型肝炎病毒X蛋白即时高表达上调LPS刺激的巨噬细胞分泌TNF-α和IL-1β。     

α-促黑素细胞激素抑制RAW264.7细胞产生炎症相关的免疫分子

目的 研究α-促黑素细胞激素（α-MSH）对巨噬细胞产生炎症相关的免疫分子的影响。探讨α-MSH的抗炎作用机制。方法 分别用LPS或α-MSH PLS处理体外培养的小鼠巨噬细胞系RAW264.7细胞,以Griess试剂测定NO,采用DADPH-黄递酶组织化学染色法检测iNOS,采用MTT法检测IL-1和TNF-α,采用ELISA法测定IL-6,细胞膜表面免疫相关分子的表达用FACS进行分析。结果 RAW264.7细胞在LPS刺激下产生NO、IL-1、IL-6和TNF-α显著增多,iNOS活性明显增强,细胞表面MHC Ⅱ、CD14、CD40、CD54、CD80和CD86分子表达水平增高;若在LPS刺激的同时给予α-MSH,能够抑制LPS诱导RAW264.7细胞产生NO,IL-1、IL-6和TNF-α,使iNOS活性及表达MHCⅡ、CD14、CD40、CD54、CD86分子降低,但CD80分子降低不明显。结论 α-MSH抑制巨噬细胞产生NO、iNOS、前炎性细胞因子及表达膜表面免疫相关分子是其抗炎作用的重要机制之一。     

小鼠ACADL对巨噬细胞表达炎症因子的影响

目的探讨长链酰基辅酶A脱氢酶(Acyl-CoA dehydrogenase,long chain,ACADL)对巨噬细胞分泌炎症因子的影响。方法在体外诱导的小鼠骨髓来源巨噬细胞中加入脂多糖(LPS)以及白细胞介素4(IL-4)分别诱导M1和M2极化,在极化0、12、24、36、48 h时用Western blot检测ACADL在M1和M2极化中的蛋白表达量。构建ACADL-pEGFP-N1质粒,使用G418筛选和鉴定过表达ACADL的RAW264.7细胞稳定株。LPS刺激过表达ACADL稳定株和对照组细胞4 h后,通过ELISA和qRTPCR检测TNF-α、IL-6、IL-10的表达情况。在RAW264.7细胞中加入beta氧化抑制因子etomoxir预处理,ELISA检测TNF-α、IL-6、IL-10的表达情况。结果在巨噬细胞极化过程中ACADL蛋白量先减少再增加;过表达ACADL的细胞中,IL-6和IL-10分泌量及mRNA水平均显著高于对照组(P0.001):beta氧化抑制因子etomoxir能够减少RAW264.7细胞分泌炎症因子。结论 ACADL能增强巨噬细胞IL-6和IL-10的分泌,其作用机制可能是通过beta氧化影响炎性细胞因子的表达。     

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.