系统性斑狼疮小鼠脾细胞中IL—12信号传递途径的研究

目的 探讨在狼疮小鼠脾细胞中是否存在IL-12通过Lck/p38/c-jun传递信号及其对脾细胞的影响。方法 以慢性移植物抗宿主病（graft versus host disease,CVHD)小鼠为狼疮小鼠模型。分别采用放射自显影、Western blot和Northern blot，检测培养的脾细胞中Lck的活性、p38磷酸化活化及c-jun基因表达的变化。结果 狼疮小鼠脾细胞经IL-12刺激后，与正常对照组相比较，Lck抑制剂PP1组Lck的活性及p38磷酸化消失，无c-jun基因的表达；加入p38抑制剂SB203580组亦无p38磷酸化及c-jun基因的表达。结果 狼疮小鼠的脾细胞异常，IL-12可通过Lck/p38/c-jun在细胞内传递信号，直接参考免疫损伤。     

系统性红斑狼疮小鼠脾细胞中IL-12信号传递途径的研究

目的探讨在狼疮小鼠脾细胞中是否存在IL-12通过Lck/p38/c-jun传递信号及其对脾细胞的影响。方法以慢性移植物抗宿主病(graft versus host disease,GVHD)小鼠为狼疮小鼠模型,分别采用放射自显影、Western blot和Northern blot,检测培养的脾细胞中Lck的活性、p38磷酸化活化及c-jun基因表达的变化。结果狼疮小鼠脾细胞经IL-12刺激后,与正常对照组相比较,Lck的活性增高、p38异常活化,c-junmRNA的水平增高。加入Lck抑制剂PP1组Lck的活性及p38磷酸化消失,无c-jun基因的表达;加入p38抑制剂SB203580组亦无p38磷酸化及c-jun基因的表达。结论狼疮小鼠的脾细胞异常,IL-12可通过Lck/p38/c-jun在细胞内传递信号,直接参与免疫损伤。     

Lck对肾小管上皮细胞的IL-12信号传递功能研究

目的 研究lck途径激活后对小鼠肾小管上皮细胞的信号传递功能的影响。方法 应用IL-12、IL-2刺激小管皮细胞,通过原位杂交检测lckmRNA在小管上皮细胞的表达;采用lck抑制剂PP1,阻抑小管上皮细胞,通过原位杂交检测lckmRNA在小管上皮细胞的表达;采用lck抑制剂PP1,阻抑小管上皮细胞的lck激酶,通过免疫印迹和放射自显影方法检测lck对小鼠上皮细胞的信号传递功能的影响。结果 lck激酶激活可调控小管上皮细胞的lck/c-Jun信号传递途径;小管上皮细胞中lck途径激活介导的细胞炎症作用与IL-12的c-Jun表达作用有关。结论 lck在IL-12的信号传递中发挥重要的作用。     

辛伐他汀通过抑制p38 MAPK活化降低肝硬化门静脉高压大鼠门静脉压力

 目的:探讨p38 MAPK信号通路在辛伐他汀降低肝硬化门静脉高压症大鼠门静脉压力（PP）中的作用。方法:采用四氯化碳复合因素法构建大鼠肝硬化门静脉高压症模型,成模后将存活大鼠随机分为模型组（n=10）、辛伐他汀治疗组（n=11）和p38 MAPK信号通路抑制剂SB203580处理组（n=10）,后2组分别给予辛伐他汀及SB203580干预处理;另设正常对照组（n=8）。处理结束后检测大鼠PP、肝脏总p38 MAPK蛋白、磷酸化p38 MAPK蛋白、总eNOS蛋白、磷酸化eNOS蛋白表达水平以及肝脏一氧化氮（NO）含量的变化。结果:（1）模型组大鼠PP明显高于正常对照组;辛伐他汀治疗组及SB203580处理组PP均明显低于模型组（P＜0.01）,辛伐他汀治疗组PP明显低于SB203580处理组（P＜0.01）。（2）与正常大鼠相比,模型组大鼠肝脏总p38 MAPK蛋白及总eNOS蛋白表达水平无明显变化（P＞0.05）,而磷酸化p38 MAPK蛋白及磷酸化eNOS蛋白表达水平分别增高与降低（P＜0.01）;辛伐他汀治疗组大鼠肝脏磷酸化p38 MAPK蛋白及磷酸化eNOS蛋白表达水平分别降低与增高（P＜0.01）;SB203580处理组大鼠肝脏磷酸化p38 MAPK蛋白及磷酸化eNOS蛋白表达水平分别降低与增高（P＜0.01）,但磷酸化eNOS蛋白表达水平增高的程度低于辛伐他汀治疗组（P＜0.01）。(3)辛伐他汀治疗组肝脏NO含量［（15.73±1.59） μmol/(g protein)］及SB203580处理组肝脏NO含量［（13.98±1.27) μmol/(g protein)］明显高于模型组［（9.81±1.12） μmol/（g protein）］（P＜0.01）,辛伐他汀治疗组NO含量明显高于SB203580处理组（P＜0.01）。结论: 辛伐他汀降低肝硬化门静脉高压症大鼠门静脉压力可能与其抑制p38 MAPK信号通路的活化有关。     

羌活提取物对哮喘小鼠Thl/Th2细胞平衡的影响及其对p38信号通路的作用

目的 探讨羌活提取物（EN）对哮喘小鼠Thl/Th2细胞平衡的影响,以及p38信号通路在其中的作用。 方法 60只雄性清洁级BABL/c小鼠,随机分为6组,每组10只,分别为正常对照组;卵蛋白(OVA)哮喘组;羌活低剂量组;羌活高剂量组、SB203580组和地塞米松组。在末次激发24 h后小鼠肺组织行HE染色。分别用ELISA检测支气管肺泡灌洗液 (BALF)中白细胞介素（IL）-4、IL-5、IL-13和γ-干扰素（IFN-γ）含量以及Western blotting检测肺组织IL-4、IFN-γ、P38蛋白表达。
结果 哮喘组小鼠与对照组小鼠相比较,BALF中炎症细胞计数、IL-4、IL-5、IL-13水平增高,而IFN-γ的表达明显降低(P<0.05),肺部炎症浸润明显(P<0.05);肺组织IL-4和磷酸化P38蛋白表达水平显著高于对照组(P<0.05),而肺组织IFN-γ的表达明显低于对照组(P<0.05)。羌活低、高剂量组、SB203580组和地塞米松组与哮喘模型组相比较,BALF中炎症细胞计数、IL-4、IL-5、IL-13浓度以及肺组织IL-4和磷酸化P38蛋白表达水平均显著降低(P<0.05);肺部炎症明显减轻(P<0.05);BALF和肺组织中IFN-γ明显升高(P<0.05)。 结论 羌活提取物具有明显的抗哮喘作用,其机制可能是通过抑制P38信号通路,影响Thl/Th2细胞平衡实现的。     

白细胞介素17通过激活p38MAPK信号通路调控人牙周膜成纤维细胞RANKL和OPG的表达

目的使用p38丝裂原活化蛋白激酶(p38MAPK)信号通路抑制剂确定白细胞介素17(IL-17)是否通过该通路参与调控人牙周膜成纤维细胞(HPDLF)核因子κB受体激活蛋白配体(RANKL)和护骨因子(OPG)的表达。方法组织块法分离培养HPDLF, 20 ng/mL IL-17分别刺激0、 20、 40、 60、 80 min,Western blot法检测HPDLF磷酸化的p38MAPK(p-p38MAPK)蛋白水平。HPDLF随机分为空白对照组、二甲基亚砜(DMSO)组、 p38MAPK抑制剂SB203580组、 IL-17组、 IL-17联合DMSO组、 IL-17联合SB203580组。IL-17及联合处理组,分别添加10μmol/L SB203580、 20 ng/mL IL-17。实时定量PCR检测HPDLF的RANKL、 OPG mRNA水平, Western blot法检测RANKL蛋白水平、 ELISA检测OPG蛋白含量。结果 p-p38MAPK蛋白水平在IL-17刺激后的0～60 min内随时间增加,在60 min时达到最高,在80 min时下降。IL-17可上调HPDLF中RANKL mRNA及蛋白表达,下调OPG mRNA和蛋白表达。较单纯使用IL-17刺激,添加SB203580通路抑制剂后, RANKL mRNA和蛋白水平均降低, OPG的mRNA水平升高。结论 IL-17通过激活p38MAPK信号通路,上调HPDLF的RANKL表达,抑制OPG mRNA表达。     

Fractalkine通过激活p38MAPK信号通路调控狼疮性肾炎小鼠Treg细胞凋亡北大核心CSCD

目的:探讨趋化因子Fractalkine(FKN)对狼疮性肾炎(LN)小鼠Treg细胞凋亡的影响及机制。方法:构建LN小鼠模型,使用FKN中和抗体(Anti-FKN)、p38MAPK信号通路的抑制剂(SB203580)、p38MAPK信号通路的激活剂(U-46619)作用于正常小鼠及模型小鼠,采用免疫组化检测小鼠肾组织中FKN、磷酸化p38(p-p38)及Foxp3的蛋白表达;采用免疫磁珠法分选模型小鼠脾脏CD4^(+)CD25^(+)Treg细胞,使用腺病毒转染细胞的方式敲低FKN的表达水平,同时使用p38MAPK信号通路抑制剂(SB203580)及激活剂(U-46619)干预细胞。采用Western blot验证FKN敲低的成功转染、检测p38、p-p38、Foxp3、凋亡相关因子(Bax、Bcl-2及Cyt-c)的蛋白表达。结果:LN小鼠肾组织中的FKN及磷酸化p38MAPK的表达明显上调,低表达FKN可降低LN小鼠肾组织及Treg细胞中FKN和p-p38的表达,增加Foxp3的表达,同时增加细胞内抗凋亡因子Bcl-2蛋白的表达,而减少促凋亡因子Bax及Cyt-c蛋白的表达。p38MAPK信号通路抑制剂对上述因子的作用与低表达FKN相似,其激活剂可以逆转这些因子在LN小鼠Treg细胞中的表达。结论:FKN可能通过激活p38MAPK信号通路参与调控LN小鼠Treg细胞凋亡,为阐明LN的发病机制提供了实验依据。     

TNF刺激肥大细胞IL-6分泌的信号转导通路

目的:检测TNF对肥大细胞IL-6、IL-10分泌和组胺释放的影响并探讨其可能的信号转导途径。方法:用不同浓度TNF激发肥大细胞系P815后收集细胞和上清,细胞用细胞激发信号ELISA(CASE)方法检测信号转导通路蛋白ERK、p38、和STAT3磷酸化水平,上清用ELISA检测组胺、IL-6和IL-10的水平。结果:ELISA结果显示TNF促进P815肥大细胞IL-6分泌(P<0.05),但对IL-10分泌和组胺释放无明显影响。ERK信号转导通路抑制剂PD98059和U0126抑制TNF引起的P815肥大细胞IL-6分泌(P<0.05),而p38信号转导通路抑制剂SB203580和STAT3信号转导通路抑制剂AG490不能抑制TNF引起的P815肥大细胞IL-6分泌。CASE结果显示ERK信号转导通路抑制剂PD98059,U0126抑制TNF引起的P815肥大细胞内ERK蛋白磷酸化(P<0.05)而p38信号转导通路抑制剂SB203580和STAT3信号转导通路抑制剂AG490不能抑制TNF引起的P815肥大细胞内p38和STAT3蛋白磷酸化。结论:TNF刺激小鼠肥大细胞P815分泌IL-6可能与ERK信号转导通路的激活有关的。     

HBeAg对小鼠骨髓源性树突状细胞成熟的影响

目的 探讨HBe Ag对LPS诱导小鼠骨髓源性树突状细胞(DC)成熟的影响。方法 体外诱导C57BL/6小鼠骨髓细胞分化成未成熟树突状细胞,经CD11c磁珠分选纯化后将DCs随机分为空白对照组、LPS刺激组、HBe Ag+LPS刺激组。流式检测DC表型变化,混合淋巴反应(MLR)检测DC促T淋巴细胞增殖能力,酶联免疫法(ELISA)检测细胞上清液中IL-12的分泌水平,Western blot检测p38磷酸化水平,并设置SB203580组为阳性对照探讨细胞IL-12分泌的可能调节机制。结果 LPS刺激未成熟DC引起细胞表面MHC-Ⅱ、CD86表达升高,刺激同种异体淋巴细胞增殖能力增强,IL-12分泌量增高。HBe Ag可抑制LPS促进DC表面MHC-Ⅱ、CD86表达升高和促淋巴细胞增殖能力增强的作用。LPS刺激DC可引起p38磷酸化水平升高,并呈时间依赖性;HBe Ag或SB203580预处理细胞再予LPS刺激,磷酸化p38表达和IL-12分泌较单纯LPS刺激组明显下降。结论 HBe Ag对LPS引起的树突状细胞的成熟有一定的抑制作用,且HBe Ag可能通过抑制p38MAPK信号通路下调LPS诱导的树突状细胞IL-12的产生。     

P38MAPK在脂多糖引起的腹膜间皮细胞损伤中的作用

目的探讨脂多糖(LPS)引起人腹膜间皮细胞(HPMC)损伤中分泌MCP-1以及LPS、P38MAPK、MCP-1三者之间可能存在的关系。方法体外培养永生化人腹膜间皮细胞(HPMC),随机分为正常对照组、LPS作用24 h组、LPS作用48 h组、特异性P38MAPK阻断剂SB203580+LPS作用24 h组、SB203580+LPS作用48 h组;Western blot法检测各组MCP-1和磷酸化P38MAPK蛋白表达水平;Real-time PCR法检测各组MCP-1 mRNA表达水平。结果 1.10 mg/L LPS刺激使HPMC的MCP-1 mRNA和蛋白质表达均较正常对照组增加(P0.05)。10 mg/L LPS作用48 h后MCP-1 mRNA和蛋白质表达水平均高于24 h组(P0.05);Western blot结果显示,与正常组对比,10 mg/L LPS作用使磷酸化P38MAPK蛋白水平明显升高(P0.05)。10 mg/L LPS作用48 h与作用24 h对比升高不明显(P0.05);经5μmol/L SB203580预处理30 min后再予以LPS刺激与单纯LPS刺激相比较,MCP-1蛋白质和mRNA均明显降低(P0.05);SB203580预处理后再予以LPS分别刺激24 h和48 h 2组相比较,MCP-1蛋白及mRNA水平差异不明显(P0.05)。结论 LPS通过磷酸化P38MAPK,导致MCP-1表达水平升高,诱发腹膜间皮细胞的损伤。     

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.