MicroRNA-146在糖尿病心肌病中的调控作用

目的:探讨微小RNA-146 (miR-146)在糖尿病心肌病(DCM)发病机制中的作用,观察miR-146及其下游靶基因表达水平的变化。方法:60只雄性C57BL/6小鼠随机分为实验(DCM)组和对照(control)组,每组30只,实验组采用低剂量(50 mg/kg链脲佐菌素(STZ)腹腔注射诱导建立糖尿病心肌病模型,对照组给予等量枸橼酸钠缓冲液腹腔注射,建模12周后取心脏做HE和Masson染色观察心脏病理改变,RT-qPCR检测miR-146 a和miR-146b及其下游靶基因白细胞介素-1受体相关激酶1(IRAK1)和TNF受体相关因子6(TRAF6)的mRNA表达,Western blot检测IRAK1和TRAF6的蛋白表达。结果:12周末HE染色显示,DCM组心肌肥大,结构紊乱;Masson染色显示,DCM组心肌内胶原纤维增多;RT-qPCR检测结果显示,DCM组中miR-146a及miR-146b较control组表达明显减少(P 0. 01),IRAK1的mRNA水平明显增高(P 0. 01),而TRAF6的mRNA水平下降(P 0. 01);Western blot检测结果显示DCM组的IRAK1蛋白表达增加,TRAF6的蛋白表达降低(P 0. 01)。结论:miR-146可能通过调节IRAK1介导的炎症反应参与糖尿病心肌损伤的发生发展。     

microRNA-146在甲亢性心脏病中的调控作用

目的探讨microRNA-146(miR-146)及其靶基因白介素1受体相关激酶(IRAK1)和TNF-α受体相关因子6(TRAF6)在甲亢性心脏病中的调控作用。方法 60只雄性C57BL/6J小鼠随机分为对照组(Control组,n=30)和甲亢性心脏病实验组(T4组,n=30)。T4组按1μg/g腹腔注射甲状腺素稀释液,Control组腹腔注射生理盐水150μL。7周后,麻醉做心电图;ELISA检测血清中TT_3及TT_4;HE和Masson染色观察心脏形态学改变;RT-qPCR检测miR-146表达;Western blot检测IRAK1及TRAF6表达。结果与Control组相比,T4组小鼠心率明显加快,心电图发现异常;血清TT_3及TT_4升高,P0.001;心肌细胞结构紊乱,胶原纤维增多;miR-146a及miR-146b表达明显上调,P0.001;IRAK1及TRAF6蛋白表达明显下调,P0.001。结论 miR-146可能通过IRAK1和TRAF6调控甲亢性心脏病。     

依托咪酯通过miR-146a 调节TLR4 通路对内毒素急性肺损伤小鼠炎症 反应的保护作用研究

目的:观察依托咪酯对内毒素急性肺损伤小鼠炎症反应的影响,并探讨其作用机制。方法:雄性成年BLAB/c小鼠60只,随机分为空白对照组、模型组、依托咪酯组,每组20只。模型组及药物干预组应用腹腔注射脂多糖(LPS,20 mg/kg)制作急性肺损伤小鼠模型,空白对照组给予等体积生理盐水腹腔注射。药物干预组于造模后0.5 h给予依托咪酯2.5 mg/kg。空白对照组及模型组给予等体积生理200μl/只。于给药后6 h检测小鼠血清中的内毒素水平,BALF中TNF-α、IL-6和IL-1β的含量;RT-PCR检测miR-146a、MyD88、IRAK1、IRF5、TRAF6 mRNA在肺组织中的表达;Western blot检测TLR4、MyD88、TRAF6、IRAK1、IRF5蛋白在肺组织中的表达。结果:模型组、依托咪酯组与空白对照组相比,血清内毒素含量、TNF-α、IL-6和IL-1β含量、miR-146a表达水平、TLR4、MyD88、IRAK1、IRF5、TRAF6 mRNA表达水平及TLR4、MyD88、IRAK1、IRF5、TRAF6蛋白表达均明显升高,差异具有统计学意义(P0.05);依托咪酯组与模型组比较,小鼠血清内毒素含量、TNF-α、IL-6和IL-1β含量、TLR4、MyD88、IRAK1、IRF5、TRAF6 mRNA表达水平及TLR4、MyD88、IRAK1、IRF5、TRAF6蛋白表达水平均明显下降,差异具有统计学意义(P0.05)。结论:依托咪酯能够通过增加miR-146a的含量调节TLR4通路对内毒素急性肺损伤小鼠炎症反应的保护作用。     

JAK2/STAT3通路参与大鼠坐骨神经结扎神经病理性疼痛模型

目的结扎大鼠双侧坐骨神经复制bCCI模型,观察JAK2/STAT3通路的激活情况。方法 60只体质量180~200 g SD雌性大鼠随机分为3组,bCCI组,sham组及Nave组。分别于bCCI手术术前1 d,术后3、7、14和21d取腰段脊髓背角,并进行RT-PCR及Western blot观察通路中主要因子的mRNA水平及蛋白水平的变化。结果 bCCI组大鼠对机械、热刺激及冷丙酮刺激的痛觉阈值明显降低。与sham组及Nave相比,bCCI组STAT3、SOCS3、JAK2及IL-6 mRNA水平显著升高,且3组在不同时间点具有差异(P0.05)。与sham组及Nave相比,bCCI组P-STAT3、JAK2和SOCS3蛋白水平升高。结论大鼠bCCI神经病理性疼痛模型JAK2/STAT3通路激活。     

小檗碱对神经病理性疼痛大鼠疼痛行为及IRAK1/TRAF6信号通路介导的神经炎症的影响

目的：探讨小檗碱对神经病理性疼痛大鼠是否具有疼痛缓解作用,并分析白细胞介素1受体相关激酶（IRAK1）/肿瘤坏死因子受体相关因子6(TRAF6)通路在其中的作用机制。方法：60只SD大鼠随机分为假手术组、脊神经结扎组（模型组）、小檗碱（低、高剂量）组及阳性药物（阿魏酸钠）组,每组12只。对各组大鼠进行动物行为学评估,测定大鼠后足热缩足反射潜伏期（TWL）、机械缩足反射阈值（MWT）;六胺银染色观察患侧脊髓背角病理变化;ELISA法检测各组大鼠背根神经节中炎症相关因子肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)和IL-6的表达水平;RT-qPCR法和Western blot法分别检测脊髓IRAK1和TRAF6的mRNA及蛋白表达水平。结果：脊髓背角病理切片显示,模型组出现明显的神经元纤维增粗并缠绕成结,且有大量空泡变性,并且神经细胞的胞质出现含有嗜银颗粒的空泡;小檗碱和阳性药物治疗后脊髓背角损伤明显减少。与假手术组相比,模型组MWT和TWL显著降低,大鼠背根神经节中TNF-α、IL-1β和IL-6含量均显著升高,脊髓中IRAK1和TRAF6 mRNA和蛋白表达量显著升高（P...     

系统性红斑狼疮患者中miR-146a表达缺陷与I型干扰素通路过度活化相关

Ι型干扰素(IFN)通路过度活化与固有免疫反应异常在系统性红斑狼疮(SLE)发病中起重要作用。miR-146a作为一个负调控因子,在固有免疫应答中发挥重要作用。为探索miR-146a在SLE发病中的作用,通过实时荧光定量聚合酶链反应技术检测18例SLE患者和11例正常对照miR-146a的表达,发现同正常对照组相比,miR-146a表达水平在SLE患者中明显降低,SLE活动组中下降则更加明显;同时检测miR-146a的靶基因IRAK1和TRAF6 mRNA表达,发现在SLE患者中其水平显著高于正常对照组;进一步分析miR-146a与IFN诱导基因表达之间的关系,结果显示两者之间存在负相关,体外实验也证实miR-146a能够负向调节I型干扰素通路。综上所述,我们的研究结果提示miR-146a可能作为一个新的生物标志物,SLE患者中miR-146a表达水平下降,导致I型干扰素通路负调控作用被削弱,从而参与疾病的发生发展。     

类风湿关节炎患者外周血miR-146a、miR-155、Ets-1以及IRAK1的表达及临床意义

为探讨类风湿关节炎(rheumatoid arthritis,RA)患者PBMC中miR-146a、miR-155、Ets-1和IRAK1的表达水平及与病情活动度的相关性,收集RA高活动期患者23例、低活动期患者21例和健康志愿者22例,采用qRT-PCR检测PBMC中miR-146a、miR-155、Ets-1、IRAK1的相对表达水平,同时分析其与临床及实验室指标的相关性。RA患者PBMC的miR-146a、miR-155相对表达量显著高于健康对照组(P<0.05),高活动组高于低活动组(P<0.05)、低活动组与健康对照组差异无统计学意义(P>0.05);RA患者PBMC的Ets-1表达水平与健康对照组差异无统计学意义,高活动组显著低于低活动组和健康对照组(P<0.05)、低活动组与健康对照组差异无统计学意义(P>0.05);RA患者PBMC的IRAK1表达水平显著低于健康对照组(P<0.05),高活动组显著低于健康对照组(P<0.01)、低活动组显著低于健康对照组(P<0.05),而高活动组与低活动组间无统计学差异(P>0.05)。在RA患者中miR-155表达水平与血沉正相关(r=0.319,P=0.042)而与血红蛋白负相关(r=-0.386,P=0.017);Ets-1与CRP负相关(r=-0.408,P=0.007);IRAK1与RF、DAS 28、总补体负相关(r=-0.513,P=0.001;r=-0.332,P=0.029;r=-0.49,P=0.015)。研究结果显示,RA高活动组患者PBMC中miR-146a、miR-155表达升高,Ets-1、IRAK1表达降低,可能参与了RA发病过程的调控;但本研究样本量有限,miR-146a、miR-155和Ets-1、IRAK1参与RA发病的关系有待后续大样本研究证实,且相关调控机制仍需进一步深入研究。     

miR-146a通过靶向调控TLR4减轻脊髓缺血再灌注大鼠损伤

目的 检测miR-146a在脊髓缺血再灌注(IR)大鼠脊髓组织的表达情况,探索miR-146a对IR大鼠炎症蛋白表达和神经功能影响的潜在机制.方法 SD大鼠通过主动脉弓夹闭14min建立IR模型,通过鞘内注射miR-146a的mimics质粒构建miR-146a过表达模型,随机分为Sham组(假手术组),IR组(缺血组...     

MyD88通路关键因子IRAK1、IRAK4和TRAF6在尖锐湿疣组织中的表达

目的观察尖锐湿疣患者皮损组织中MyD88通路关键因子白介素-1受体相关激酶-1(IRAK1)、白介素-1受体相关激酶-4(IRAK4)和肿瘤坏死因子受体相关因子6(TRAF6)的表达。方法选取60例尖锐湿疣患者作为观察组,60例正常包皮环切者作为对照组,免疫组化法检测皮损组织中MyD88通路关键因子IRAK1、IRAK4和TRAF6的表达水平,并进行临床分析。结果IRAK1、IRAK4和TRAF6在观察组中普遍呈高表达,所占比例分别为80%、83.3%和91.7%,对照组中无表达或呈低表达,所占比例分别为96.7%、98.3%和100%,差异均有统计学意义(P0.05)。结论尖锐湿疣患者皮损组织中MyD88通路关键因子IRAK1、IRAK4、TRAF6的表达显著高于正常对照组。     

miR-146a在大鼠缺血再灌注肝损伤中的表达及作用

目的:评价肝组织微小RNA-146a(miR-146a)表达变化与大鼠缺血再灌注(I/R)炎性肝损伤的关系。方法:72只SD大鼠随机分为非手术组(N组)、假手术组(S组)及I/R组,按分组处理后分别检测各组大鼠0、2、12和24 h(每组各时点n=6)血浆丙氨酸转氨酶(ALT)活性及白细胞介素6(IL-6)和肿瘤坏死因子α(TNF-α)含量,real-time PCR检测肝组织Toll样受体4(TLR4)、IL-6、TNF-α和miR-146a表达,Western blot分析肝组织中TLR4、IL-6和TNF-α表达。结果:N组大鼠各时点血浆ALT活性及IL-6和TNF-α含量与肝组织中TLR4、IL-6和TNF-α表达量无显著差异。S组大鼠血浆IL-6和TNF-α含量升高(P0.01),但血浆ALT活性与肝组织TLR4、IL-6和TNF-αm RNA及蛋白的表达量却无明显变化。I/R后,大鼠血浆ALT活性及IL-6和TNF-α含量明显升高(P0.01),肝组织TLR4、IL-6和TNF-αm RNA及蛋白的表达量亦显著升高(P0.01),且随着时间延长,各指标仍持续升高;肝组织miR-146a表达量显著下降,其中在缺血12 h时下降最为明显,到24 h表达量再次升高,但始终低于I/R前(P0.01)。N组及S组大鼠各时点肝组织miR-146a表达量显著高于I/R组I/R后相应时点的表达(P0.01)。结论:I/R大鼠肝组织miR-146a表达下降的同时存在TLR4信号通路的激活及炎性肝损伤的发生。     

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.     

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.