FIZZ1在ApoE基因敲除小鼠动脉粥样硬化斑块内的表达

目的:探讨FIZZ1在ApoE基因敲除小鼠粥样斑块内的表达。方法:C57BL/6JApoE基因敲除鼠及C57BL/6J野生型小鼠各9只,分别喂食高脂饲料及普通饲料,24周后处死,自主动脉根部至腹主动脉离断整支血管,石蜡包埋后作连续切片,行HE染色及FIZZ1免疫组化,检测血管斑块内FIZZ1表达情况,RT-PCR检测斑块内FIZZ1mRNA表达。结果:ApoE基因敲除鼠高脂饲养24周后,主动脉根部动脉粥样硬化明显,斑块体积较大,免疫组化及其RT-PCR可见FIZZ1及其mRNA在粥样硬化斑块内明显表达,同龄野生型C57BL/6J鼠血管壁内未见FIZZ1及其mRNA表达。结论:FIZZ1能在动脉粥样硬化斑块内表达。     

炎症反应通过上调CD36表达促进小鼠肾脏损伤

目的:探讨慢性炎症对小鼠肾脏CD36表达的影响及其在小鼠肾脏损伤中的作用。方法:将8周龄雄性C57BL/6J小鼠和CD36基因敲除(CD36KO)小鼠随机分为C57BL/6J生理盐水注射组、C57BL/6J酪蛋白注射组和CD36KO酪蛋白注射组,每组8只。高脂喂养处理14周后,收集小鼠血清、24 h尿液和肾组织样本。ELISA试剂盒检测血清中肿瘤坏死因子α(TNF-α)含量,全自动生化仪测定血、尿肾功能指标,HE染色和Masson染色分析肾脏病理改变,real-time PCR和Western blot检测肾脏组织中CD36及炎症/趋化因子(MCP-1、IL-6和TNF-α)m RNA和蛋白的表达,试剂盒测定组织内过氧化氢含量,免疫组化染色测定肾组织Nrf2和TGF-β1的蛋白表达。结果:与生理盐水注射组相比,酪蛋白注射能增强C57BL/6J小鼠血清TNF-α含量和肾组织中TNF-α的蛋白表达(P 0. 05),提示酪蛋白注射能成功诱导小鼠全身和肾脏局部的慢性炎症。同时酪蛋白注射显著促进了小鼠肾组织的CD36和TGF-β1蛋白表达,引起肾小球硬化、蛋白尿和血清肌酐含量显著增加,组织过氧化氢含量明显增加,Nrf2含量和抗氧化能力明显降低(P 0. 05)。而酪蛋白处理的CD36基因敲除小鼠肾组织病理学改变不明显,血、尿肾功能指标和尿量较酪蛋白处理的C57BL/6J小鼠明显降低,且肾组织过氧化氢含量低于酪蛋白处理的C57BL/6J小鼠(P 0. 05)。结论:炎症应激通过促进小鼠肾组织CD36表达,促进氧化应激,导致小鼠肾损伤。     

瞬时受体电位通道(TRPV1)在热性惊厥中的作用及其机制研究

<正>目的:通过分析C57BL/6野生型小鼠和TRPV1基因敲除鼠热性惊厥行为学、脑内炎性因子表达变化,探讨TRPV1对热性惊厥发生及复发的致病机制。方法:高热诱导C57BL/6野生型小鼠和TRPV1基因敲除鼠热性惊厥后:比较两组间热性惊厥行为学、脑电图及核心温度的差异;ELISA检测两组间大脑及经辣椒素和高热处理后BV2细胞中炎性因子的表达差异。结果:     

利用CRISPR/Cas9技术制备成对框基因2敲除小鼠

目的 利用成簇规律间隔短回文重复序列（CRISPR）/重组CRISPR相关核酸酶9（Cas9）技术双切口法制备成对框基因2（Pax2）敲除小鼠,为探讨Pax2基因在多个系统发育的作用提供动物模型。方法 根据Pax2基因序列设计sgRNA,设计出的sgRNA和Cas9体外转录后显微注射到C57BL/6J 小鼠的受精卵中,F0代小鼠出生后取其基因DNA测序鉴定基因型。共获得8只F0 代小鼠,使敲除成功的F0代小鼠与野生C57BL/6J 小鼠交配,获得F1代小鼠,后均采用基因成功敲除的小鼠与C57BL/6J 小鼠进行交配,可获得稳定的Pax2基因敲除小鼠。结果 成功获得可稳定繁殖的Pax2杂合子基因敲除小鼠,其Pax2基因缺失1628 bp;组织HE染色显示,敲除小鼠的肾小球数量明显减少;Western blotting结果显示,敲除小鼠的肾皮质Pax2蛋白表达较野生型小鼠减少。结论 利用CRISPR/Cas9技术可成功构建Pax2杂合子基因敲除小鼠,为进一步研究Pax2基因的作用奠定基础。     

近端小管RAS功能轴在早期糖尿病小鼠肾脏中的变化

目的：观察自发性糖尿病小鼠模型中，糖尿病肾病早期近端小管肾素-血管紧张素系统（RAS）两条轴[血管紧张素转换酶（ACE）-血管紧张素II(Ang II)-血管紧张素II 1型受体（AT1）经典轴和ACE2-Ang(1-7)-Mas受体（MasR）新轴]的变化特征及对肾脏结构和功能的影响。方法：应用雄性自发性1型糖尿病Akita小鼠（以C57BL/6背景的雄性非糖尿病小鼠为正常对照）和自发性2型糖尿病db/db小鼠（以C57BLKS/J背景的雄性db/m非糖尿病小鼠为正常对照），饲养至16周龄时处死小鼠，测定相应生理指标。收集尿液，检测尿液中Ang II和Ang(1-7)的含量；收取肾脏组织行病理切片染色观察肾脏结构变化；免疫组化法观察肾脏组织血管紧张素原（Agt）、ACE、ACE2和MasR的表达部位及变化；Western blot和RT-qPCR测定近端肾小管的上述蛋白及mRNA的变化。结果：两种糖尿病小鼠16周龄时，肾小球滤过率和尿白蛋白/肌酐比值显著增加（P<0.05）,PAS染色可见近端肾小管细胞体积增加，管腔扩张，肾小管肥大，肾小管损伤分数显著增加（P<0.01）...     

高同型半胱氨酸血症与胱硫醚-β-合酶敲除小鼠紫癜性肾炎加重的相关性

目的 研究高同型半胱氨酸血症（HHcy）与胱硫醚-β-合酶（CBS）敲除（KO）小鼠紫癜性肾炎（HSPN）加重的相关性。方法 选择CBS-野生型（WT）雄性C57BL/6J小鼠、雄性CBS-KO小鼠（C57BL/6J背景）各40只，鼠龄8～10周，体质量18～20 g。CBS-WT小鼠和CBS-KO小鼠均采用印度墨水尾静脉注射联合麦醇溶蛋白灌胃建立HSPN模型，造模过程中给予常规饮食或高蛋氨酸饮食，给予对照溶剂、内质网应激（ERS）激动剂或拮抗剂腹腔注射。检测血清同型半胱氨酸（Hcy）含量、血肌酐（SCr）、尿素氮（BUN）、24 h尿蛋白、肾脏病理改变及肾小球内细胞凋亡率、C/EBP同源蛋白（CHOP）、含半胱氨酸的天冬氨酸蛋白水解酶（caspase）-12、caspase-3表达水平。结果 常规饮食CBS-WT+HSPN组、高蛋氨酸饮食CBS-WT+HSPN组均出现肾脏病理改变，SCr、BUN、24 h尿蛋白水平高于常规饮食CBS-WT组（P <0.05），且常规饮食CBS-WT+HSPN组与高蛋氨酸饮食CBS-WT+HSPN组肾脏病理改变及SCr、BUN、24 h尿蛋白水平...     

异基因MHC Ⅰ修饰对骨髓细胞的免疫学特性的影响

目的：观察异基因MHC Ⅰ修饰对骨镑细胞的免疫学特性的影响，探讨MHC Ⅰ类分子在诱导免疫耐受中的作用机制。方法：由逆转录病毒载体pMSCV介导，将BALB/C小鼠的MHC Ⅰ类分子H—2D^d基因导入C57BL/6小鼠的骨骼细胞，流式细胞仪检测转染后基因的表达。MTT法检测混合淋巴细胞反应，乳酸脱氢酶释放法测定BALB/C小鼠NK细胞对H-2D^d修饰后C57BL/6小鼠骨髓细胞的杀伤活性。结果：重组逆转录病毒感染的C57BL/6小鼠骨髓细胞对BALB/C小鼠脾细胞的刺激强度或应答程度，与未转染和空载体病毒感染的C57BL/6小鼠骨髓细胞相比都显著减弱。BALB/C小鼠NK细胞对转染H-2D^d基因后的C57BL/6小鼠骨髓细胞的杀伤显著降低。结论：在骨髓移植中用受者MHC Ⅰ分子修饰供者骨髓细胞可能诱导免疫耐受。     

FGL2拮抗新型冠状病毒鼠株MA17感染北大核心CSCD

目的通过新冠病毒小鼠适应性毒株MA17感染老年WT C57BL/6和Fgl2基因敲除小鼠,鉴定Fgl2基因敲除是否可以拮抗MA17毒株感染。方法使用MA17毒株感染老年WT小鼠和Fgl2基因敲除小鼠,记录生存率及体质量变化,通过H&E染色、免疫组化及Masson染色鉴定感染小鼠的炎性浸润及纤维化形成。结果与WT对照小鼠相比,Fgl2基因敲除小鼠表现出显著的生存率延长,体质量无明显下降,肺组织纤维蛋白原沉积减少以及肺纤维化显著改善。结论Fgl2基因敲除小鼠可以拮抗新型冠状病毒鼠株MA17感染引起的血栓及纤维化形成。     

丹参多酚酸盐对ApoE基因敲除小鼠血清TNF-α的影响

目的:观察中药丹参多酚酸盐(salvianolate)对C57BL/6JApoE基因敲除小鼠血清肿瘤坏死因子(tumor necrosis factor-α, TNF-α)的影响,以探讨其抗动脉粥样硬化的可能机制.方法:给8周龄雄性C57BL/6J ApoE基因敲除小鼠喂食高脂饮食,腹腔注射丹参多酚酸盐,随机将其分为模型组(仅腹腔注射生理盐水)、丹参多酚酸盐低剂量(60mg/kg)组、中剂量(120mg/kg)组、高剂量(240mg/kg)组,每组12只,共50只;正常对照组(即C57BL/6J野生型小鼠)10只.32周末时处死各组小鼠,留取血清,采用放射免疫分析检测各组小鼠血清TNF-α水平.结果:32周末时,随丹参多酚酸盐剂量的增加,ApoE基因敲除小鼠血清TNF-α浓度逐渐降低,丹参多酚酸盐各剂量组血清TNF-α水平均明显低于模型组(P均＜0.01);丹参多酚酸盐低剂量组与正常对照组比较,无统计学意义(P＞0.05),余两组与正常组比较,有明显统计学意义(P均＜0.01);丹参多酚酸盐各浓度组之间亦有明显差异(P均＜0.01).结论:各剂量组丹参多酚酸盐均能够降低ApoE基因敲除小鼠血清TNF-α水平,随剂量增加,减低越明显,说明丹参多酚酸盐能够抑制动脉粥样硬化炎症反应,可能是其抗动脉粥样硬化的作用机制之一.     

AQP3 缺失小鼠尿素和尿浓缩能力的研究

目的： 肾脏表达水通道蛋白3（AQP3）在尿浓缩机制中起着极其重要的作用。AQP3缺失小鼠表现为尿浓缩功能严重障碍。为阐明这种基因缺失的生理特性和发生机制,我们利用AQP3基因敲除小鼠和野生型小鼠复制急性尿素负荷动物模型,对其作用机制及影响进行研究。方法: 对小鼠实施急性尿素负荷,从实验前2 h到尿素负荷后第8 h,每隔2 h收集1次尿样分别检测尿量、尿渗透压及尿素浓度。分离肾脏组织RNA进行实时荧光定量RT-PCR反应。应用Western blotting分析肾组织中尿素转运蛋白（UTs）的变化。结果: 给予尿素后4 h左右,2种不同基因型小鼠均有负荷尿素的排泄。尿素负荷后AQP3缺失小鼠尿的渗透压和尿素含量逐渐增高,第 8 h,几乎与野生型小鼠值相等,但尿中非尿素溶质浓度却没有改变。在最后4 h内尿量下降到基础值的1/4。AQP3基因缺失和野生型小鼠尿素负荷显著地上调了UT-A3的表达。结论: AQP3基因缺失并没有直接干扰肾脏对尿素的浓缩功能,但却减弱了对尿中其它溶质浓缩的能力。这种对溶质选择性的反应是由于AQP3对水和尿素转运能力的不同所致。结果表明AQP3对尿中非尿素溶质的浓缩具有特殊作用。     

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.     

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.     

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.