Systemic amyloid A (AA) amyloidosis, which is considered the second most common form of systemic amyloidosis usually takes place several years prior to the occurrence of chronic inflammation, generally involving the kidney. Activated HSF1, which alleviated unfolded protein response (UPR) or enhanced HSR, is the potential therapeutic target of many diseases. However, the effect of HSF1 on AA amyloidosis remains unclear. This study focused on evaluating effect of HSF1 on AA amyloidosis based on HSF1 knockout mice. As a result, aggravated amyloid deposits and renal dysfunction have been found in HSF1 knockout mice. In progressive AA amyloidosis, HSF1 deficiency enhances serum amyloid A production might to lead to severe AA amyloid deposition in mice, which may be related to deactivated unfolded protein response as well as enhanced inflammation. Thus, HSF1 plays a significant role on UPR related pathway impacting AA amyloid deposition, which can mitigate amyloidogenic proteins from aggregation pathologically and is the possible way for intervening with the pathology of systemic amyloid disorder. In conclusion, HSF1 could not only serve as a new target for AA amyloidosis treatment in the future, but HSF1 knockout mice also can be considered as a valuable novel animal model for renal AA amyloidosis.

病理学是基础医学课程中主要的医学学科之一,是基础医学和临床医学之间的桥梁.它主要立足于组织器官的形态学改变,从病因、发病机制、病理改变、疾病的结局和转归等方面研究疾病的发生和发展规律.随着近年来信息技术的飞速发展,同时具有线上课程和线下课堂教学优势的线上线下混合式教学也越来越多地应用于医学教学中.本文突破传统课堂教学式授课方式,希望能找到一种更有效的方法来提高病理学实验教学的效果.

目的:探讨思维导图结合翻转课堂的教学方法对病理教学的影响.方法:以试卷测评和问卷调查的形式,评价不同教学方法在病理教学中的效果.结果:应用思维导图结合翻转课堂的实验组学生成绩明显优于对照组(P＜0.01);并且80％以上的学生认可新的教学方法.结论:思维导图结合翻转课堂的教学方法对病理教学效果有显著的促进作用.

目的 探讨HSP70-TLR4-MyD88信号通路在肾脏AA淀粉样变中的作用机制,以及Hsf1基因对其通路的调节作用.方法 使用Hsf1基因敲除小鼠构建肾脏AA淀粉样变模型,以野生型小鼠为对照.模型制备成功后,从各组小鼠肾脏组织中提取mRNA进行检测评估;使用刚果红和免疫组化染色比较各组肾脏AA淀粉样物质的沉积程度.构建替普瑞酮(GGA)治疗组模型,对比治疗组与实验组肾脏淀粉样变情况.结果 淀粉样变诱导试验后,Hsf1+/+小鼠肾脏HSP70 mRNA表达量显著升高,而Hsf1-/-小鼠肾脏组织HSP70 mRNA表达量无明显改变.Hsf1-/-淀粉样变诱导组小鼠肾脏TLR4及下游因子MyD88以及细胞因子Tnf-α和IL-6的mRNA表达量均显著高于Hsf1+/+淀粉样变诱导组小鼠.刚果红染色和免疫组化染色示:Hsf1-/-淀粉样变诱导组肾脏淀粉样物质沉积程度明显高于Hsf1+/+淀粉样变诱导组(P<0.01),其余生理盐水组肾脏未见淀粉样沉积.给予GGA治疗后,有效诱导了HSP70表达,从而抑制了炎症刺激后炎症因子的高表达,并一定程度上削弱了淀粉样变沉积程度.结论 Hsf1通过调控HSP70-TLR4-MyD88信号通路有效抑制了肾脏AA淀粉样变,为淀粉样变疾病的临床预防与治疗提供了新思路.

A蛋白沉积型(AA)淀粉样变,是由淀粉样前体蛋白发生错误的折叠聚集,形成不溶性纤维沉积于多种组织器官内致使其功能障碍而引起.肾脏是该疾病最常出现并受累的重要脏器,其发病多见于中老年,早期易出现各种误诊、漏诊,同时该疾病病死率高,并且预后较差.因此,深入了解该疾病特点将有助于临床诊断和治疗策略的制定.本文将从发病机制、疾病谱、临床诊断以及治疗策略等方面对肾脏AA淀粉样变的研究进展进行综述.

目的 探讨ATF6-XBP1通路在肝脏AA淀粉样变中的作用机制,以及HSF1对其通路的调节作用.方法 使用HSF1基因敲除小鼠构建肝脏淀粉样变模型,以野生型为对照组.采集各组小鼠肝脏组织,通过刚果红和免疫组化SP法比较各组肝脏淀粉样变沉积程度,同时对ATF6和XBP1的mRNA及蛋白水平进行检测.结果 刚果红染色和免疫组化SP法染色:敲除型实验组肝脏淀粉样物质沉积程度显著高于野生型实验组(P＜0. 01),敲除型空白组、野生型空白组肝脏未见淀粉样沉积.Western blot结果显示,敲除型实验组肝脏中GRP78蛋白表达量显著低于野生型实验组.RT-PCR结果显示:野生型实验组ATF6mRNA表达略低于野生型空白组,前者的XBP1mRNA表达则显著高于后者;而在敲除型组中ATF6、XBP1mRNA表达呈相反趋势.结论 HSF1通过调控ATF6-XBP1通路影响肝脏AA淀粉样变的形成与沉积,证实热休克反应与未折叠蛋白质反应的相互作用,为淀粉样变疾病的临床预防与治疗提供新的理论依据.

目的:探讨热休克转录因子1(HSF1)在心脏淀粉样变疾病中的作用机制.方法:将30只HSF1敲除小鼠随机分为3个实验组,HSF1敲除型空白对照组、敲除型淀粉样变模型组及替普瑞酮治疗组;将20只野生型小鼠随机分为2个实验组,野生型空白对照组、野生型淀粉样变模型组,每组均为10只.进行淀粉样变诱导实验构建淀粉样变疾病模型.模型构建2个月后处死小鼠,通过刚果红染色、免疫组织化学染色、分子生物学检测,比较各个实验组心脏组织中热休克蛋白及淀粉样变沉积情况,以及肝脏、脾脏、胃、肠舌、皮肤组织中的淀粉样变沉积情况. 结果:淀粉样变诱导实验后,野生型小鼠心脏组织中热休克蛋白70(HSP70)mRNA的表达水平较空白组显著升高(P＜0.05),HSF1敲除小鼠淀粉样变疾病模型的心脏组织中HSP70 mRNA的表达水平显著低于野生型实验组(P＜0.05),且心脏淀粉样变沉积程度显著高于野生型实验组.给予HSP70诱导剂替普瑞酮后,敲除型实验组心脏组织中的HSP70 mRNA的表达水平明显升高,其淀粉样沉积程度明显得到改善.而未进行淀粉样变诱导实验的空白组,不管是野生型小鼠还是敲除型小鼠均未见淀粉样物质沉积.此外,敲除型实验组小鼠血清中心肌肌钙蛋白T(TnT)表达显著高于野生型实验组,而给予替普瑞酮治疗后表达下降. 结论:HSF1可能通过对HSP70的调控影响心脏淀粉样变的沉积情况,该结果为心脏淀粉样变的防治提供了新的思路.

Objective To establish and characterize a cardiac amyloidosis mouse model. Methods In this study, senile systemic amyloidosis was induced through injecting amyloid fibrils into female Hsf1-/-knockout mice or wide type mice. Two months after the injection, tissue amyloid deposition and cardiac functions were assessed in experimental mice by Congo red staining, immunohistochemistry and transmission electron microscopy. Results Two months after amyloidosis induction, the levels of tissue amyloid deposits were similar between Hsf1-/- and wild-type mice, except in heart where Hsf1-/- showed significantly higher level of deposition. Cardiac amyloids mainly deposited in the connective tissue and small vessel wall of myocardium, the extend of which correlated with cardiac hypertrophy and contractile dysfunction. Furthermore, Hsf1-/- knockout mice exhibited increased cardiac AApoAII deposition. Conclusion These results suggested that Hsf1-/-knockout mice could be used as a senile cardiac amyloidosis model for relevant study.

Amyloidosis is a severe systemic disease which is a protein conformational disorder characterized by extracellular accumulation of amyloid fibrils derived from various proteins [ 1 Merlini G. Westermark P. The systemic amyloidoses: clearer understanding of the molecular mechanisms offers hope for more effective therapies. J. Intern. Med. 2004; 255: 159-178 Crossref PubMed Scopus (178) Google Scholar , 2 Sipe J.D. Cohen A.S. Review: history of the amyloid fibril. J. Struct. Biol. 2000; 130: 88-98 Crossref PubMed Scopus (868) Google Scholar ]. Cardiac involvement may occur in the three main types of amyloidosis: acquired monoclonal light-chain, hereditary transthyretin and senile amyloidosis, and has a major impact on prognosis. In mouse senile amyloidosis, apolipoprotein A-II (apoAII)—a constituent of serum high-density lipoprotein—accelerates age-associated deposited extracellularly as amyloid fibrils (AApoAII) in the heart, liver, kidney, intestine, skin, tongue and other organs except the brain [ [3] Higuchi K. Kitagawa K. Naiki H. Hanada K. Hosokawa M. Takeda T. Polymorphism of apolipoprotein A-II (apoA-II) among inbred strains of mice. Relationship between the molecular type of apoA-II and mouse senile amyloidosis. Biochem. J. 1991; 279: 427-433 Crossref PubMed Scopus (91) Google Scholar ]. Heat shock factor 1 (HSF1) binds to DNA at a specific site of Hsp gene promoter regions known as the heat shock element and regulates the activation of the canonical pathway [ [4] Young J.C. Agashe V.R. Siegers K. Hartl F.U. Pathways of chaperone-mediated protein folding in the cytosol. Nat. Rev. Mol. Cell Biol. 2004; 5: 781-791 Crossref PubMed Scopus (952) Google Scholar ], which is required for the acquisition of thermotolerance and protection of cells from degenerative diseases [ [5] Fujimoto M. Nakai A. The heat shock factor family and adaptation to proteotoxic stress. FEBS J. 2010; 277: 4112-4125 Crossref PubMed Scopus (169) Google Scholar ]. Moreover, it is well known that HSF1 and Hsps activities decline during the aging process [ [7] Calderwood S.K. Murshid A. Prince T. The shock of aging: molecular chaperones and the heat shock response in longevity and aging — a mini-review. Gerontology. 2009; 55: 550-558 Crossref PubMed Scopus (247) Google Scholar ], while a reduction in amyloid fibril aggregation can decelerate aging and increase the life-span of Caenorhabditis elegans and mice [ [8] Cohen E. Bieschke J. Perciavalle R.M. Kelly J.W. Dillin A. Opposing activities protect against age-onset proteotoxicity. Science. 2006; 313: 1604-1610 Crossref PubMed Scopus (706) Google Scholar ]. Although experiments in cells and animal models have shown that increased levels of molecular chaperones such as HSPB1, DNAJB1, and HSPA1A can inhibit protein aggregation in Parkinson's, Huntington's, and polyglutamine diseases, however there was no strong evidence supporting the correlation between HSF1 and/or Hsps and cardiac amyloidosis.

目的:探讨载脂蛋白A-Ⅱ形成的小鼠老化淀粉样纤维是否可形成并沉积于骨骼肌中.方法:采用R1.P1-Apoa2c小鼠进行淀粉样变诱导实验,诱导实验进行2个月后,通过组织学、免疫组织化学、透射电镜及分子生物学检测骨骼肌中淀粉样纤维的沉积程度.结果:通过淀粉样变诱导实验2个月及4个月后,淀粉样纤维主要沉积于小鼠骨骼肌中的结缔组织及肌内膜内.结论:小鼠老化淀粉样纤维可在骨骼肌肌内膜及结缔组织中沉积,并为淀粉样变疾病的发生机制提供了新的理论依据.

Mouse senile amyloidosis is a disorder in which apolipoprotein A-II deposits extracellularly in many organs as amyloid fibrils (AApoAII). In this study, we intravenously injected 1 μg of isolated AApoAII fibrils into R1.P1-Apoa2c mice, to induce AApoAII amyloidosis. We observed that the unfolded protein response was induced by deposition of AApoAII amyloid. We found that the mRNA and the protein expression levels of heat shock protein A5 (HSPA5; also known as glucose-regulated protein 78) were increased in the liver with AApoAII amyloid deposits. Immunohistochemistry showed that HSPA5 was only detected in hepatocytes close to AApoAII amyloid deposits. Furthermore, gene transcription of several endoplasmic reticulum (ER) stress-related proteins increased, including eukaryotic translation initiation factor 2 alpha kinase 3 (Eif2ak3), activating transcription factor 6 (Atf6), activating transcription factor 4 (Atf4), X-box-binding protein 1 splicing (Xbp1s), DNA-damage inducible transcript 3 (Ddit3), and autophagy protein 5 (Atg5). Moreover, apoptosis-positive cells were increased in the liver. Similar results were seen in the kidney but not in the heart. Our study indicates that ER stress responses differed among tissues with extracellular AApoAII amyloid fibril deposition. Although upregulated HSPA5 and the activated unfolded protein response might have roles in protecting tissues against aggregated extracellular AApoAII amyloid deposition, prolonged ER stress induced apoptosis in the liver and the kidney.

ABSTRACT:Recently ,prion‐like transmission has been found in various amyloidosis .AApoAII amyloid fibrils in mouse senile amyloidosis have exhibited transmissibility .AApoAII amyloid fibrils ,which were excreted from mice and contained in fe‐ces or milk ,cause mouse senile amyloidosis .However ,transmissibility of AApoAII amyloid fibrils through other pathways has not yet been established .In this study ,we injected AApoAII amyloid fibrils into R1 .P1‐A poa2c mice to induce AApoAII sys‐temic amyloidosis .Two months later ,AApoAII amyloid fibrils ,which deposited in the skeletal muscles of amyloid‐affected mice ,were used to induce AApoAII systemic amyloidosis .Mouse senile amyloidosis which deposited in skeletal muscles could induce secondary transmission of AApoAII amyloidosis .The evidence of transmission through skeletal muscles in non‐prion systemic amyloidosis is found in our study .This pathway of transmission provides new insight into the potential for food‐borne pathogenesis and etiology of systemic amyloidosis .

探讨“以问题为导向的教学方法（problem—basedlearningPBL），在病理教学中的应用，经一学期的实践，我们体会到PBL教学法的优点在于：充分调动学生学习的积极性，变被动学习为主动学习，提高学生自学的能力。同时，它要求教师必须扩大知识面；尤其我们青年教师既要精通本专业的知识和发展前沿，又要加强相关科知识的学习和研究。因此，PBL教学法是一种教学相长的好方法。

Objective To study the effects of rehabilitation training on the synaptic ultrastructure of the brain cortex after cerebral ischemia.Methods Sixty rats were randomly divided into a sham operation group,a cerebral ischemia group and a rehabilitation training group.A model of focal cerebral ischemia was created in the rats of the cerebral ischemia and rehabilitation training groups.The rehabilitation training group was given balancing,grasping,rotation and walking training every day.The sham operation group and cerebral ischemia group were kept in normal cages,taking food and moving freely.Electron microscopy was used to observe the density and ultrastructure of the cortical synapses at days 1,7,14 and 21 after the operation.Results In the sham operation group there were numerous synaptic terminals in the cortical neuropil.The majority of synapses were Gray type Ⅱ,containing spherical synaptic vesicles in presynaptic elements.After cerebral ischemia,the number of synaptic terminals had decreased and the synaptic structure was damaged.The synaptic vesicles lessened and dissolved.The presynaptic and postsynaptic membranes became unclear.At 21d after ischemia,the synaptic vesicles had disappeared and the presynaptic and postsynaptic membranes were damaged so seriously that the typical synaptic structure disappeared.After rehabilitation training,the damaged synaptic terminals recovered.Conclusions The synaptic structure was damaged after cerebral ischemia,and the damaged synaptic terminals recovered after rehabilitation training.Rehabilitation training can enhance synaptogenesis in the brain's cortex after cerebral ischemia and promote the recovery of motor function,at least in rats.

Apolipoprotein A-II (apoA-II) is the second major apolipoprotein following apolipoprotein A-I (apoA-I) in HDL. ApoA-II has multiple physiological functions and can form senile amyloid fibrils (AApoAII) in mice. Most circulating apoA-II is present in lipoprotein A-I/A-II. To study the influence of apoA-I on apoA-II and AApoAII amyloidosis, apoA-I-deficient (C57BL/6J.Apoa1−/−) mice were used. Apoa1−/− mice showed the expected significant reduction in total cholesterol (TC), HDL cholesterol (HDL-C), and triglyceride (TG) plasma levels. Unexpectedly, we found that apoA-I deficiency led to redistribution of apoA-II in HDL and an age-related increase in apoA-II levels, accompanied by larger HDL particle size and an age-related increase in TC, HDL-C, and TG. Aggravated AApoAII amyloidosis was induced in Apoa1−/− mice systemically, especially in the heart. These results indicate that apoA-I plays key roles in maintaining apoA-II distribution and HDL particle size. Furthermore, apoA-II redistribution may be the main reason for aggravated AApoAII amyloidosis in Apoa1−/− mice. These results may shed new light on the relationship between apoA-I and apoA-II as well as provide new information concerning amyloidosis mechanism and therapy.

Objective:To explore the effect of chronic intermittent hypobaric hypoxic(CIIH) preconditioning on the expression of c-Fos and Bcl-2 in hypoxia injured parietal cortex of rats.Methods:Fourty-eight adult healthy male SD rats were randomly divided into control group(CON),chronic intermittent hypobaric hypoxia group(CIIH),acute hypoxia group(AH) and hypoxic preconditioning group(CIIH+AH).Toluidine blue staining,immunohistochemistry technique and transmission electron microscope were used to monitor the expression of c-Fos and Bcl-2 in parietal cortex.Results:No obvious neural denaturation in parietal cortex was found in CIIH group.Between the two groups of CON and CIIH,there was no statistical difference(P>0.05).Compared with CON group and CIIH group,neural serious denaturation in parietal cortex was found in AH group.The expression of c-Fos and Bcl-2 in AH group was significantly increased(P<0.01).Compared with AH group,the neurons in CIIH+AH group were more slightly injured,the expression of Bcl-2 was increased(P<0.05),while the expression of c-Fos was reduced(P<0.05).Conclusion:CIIH preconditioning may inhibit hypoxia-induced apoptosis by down-regulation of c-Fos expression and up-regulation of Bcl-2 expression,hereby protecting the brain.

Amyloidosis -Mechanisms and Prospects for Therapy 164 species, AApoAII and AA amyloidosis occurs sporadically and is associated with aging (AApoAII amyloidosis) and chronic inflammation (AA amyloidosis).However, intriguing recent data suggest that both AApoAII and AA amyloidoses could be transmitted by a prion-like infectious process through a seeding-nucleation mechanism (Qian et al., 2010;B. Zhang et al., 2008).In these amyloidoses, AApoAII and AA amyloid fibrils, abnormal forms of the host serum proteins (apoA-II and SAA), induce conformational changes in apoA-II and SAA to form AApoAII and AA fibrils, and this causes detectable phenotypes or diseases in affected individuals.Recently, Aß amyloid fibrils in Alzheimer's disease and intracellular amyloid fibril-like aggregated proteins were postulated to work as seeds for propagation of mis-folded and pathologic protein structures in various neurodegenerative disorders, including Huntington's disease, Parkinson's disease and tauopathies (Aguzzi et al., 2009;Brundin et al., 2010).The prion-like transmission of amyloid fibrils or fibril-like materials, which could play an important role in the propagation of pathological events in systemic amyloidosis, will be discussed here.

Patients on long-term hemodialysis can develop dialysis-related amyloidosis (DRA) due to deposition of β2-microglobulin (β2m) into amyloid fibrils (Aβ2M). Despite intensive biochemical studies, the pathogenesis of amyloid deposition in DRA patients remains poorly understood. To elucidate the mechanisms that underlie Aβ2M fibril formation in DRA, we generated transgenic mice that overexpress human β2m protein in a mouse β2m gene knockout background (hB2MTg+/+ mB2m+/+). The hB2MTg+/+mB2m−/− mice express a high level of human β2m protein in many tissues as well as a high plasma β2m concentration (192.8 mg/L). This concentration is >100 times higher than that observed in healthy humans and >4 times higher than that detected in patients on dialysis. We examined spontaneous and amyloid fibril-induced amyloid deposition in these mice. Amyloid deposition of β2m protein was not observed in aged or amyloid fibril injected animals. However, mouse senile apolipoprotein A-II amyloidosis (AApoAII) was detected, particularly in the joints of mice that were injected with AApoAII amyloid fibrils. This study demonstrates that this mouse model could be valuable in studying the components and conditions that promote DRA, and indicates that high plasma concentrations of hβ2m as well as seeding with pre-existing amyloid fibrils may not be sufficient to induce Aβ2M.

Amyloidosis describes a group of protein folding diseases in which amyloid proteins are abnormally deposited in organs and/or tissues as fine fibrils. Mouse senile amyloidosis is a disorder in which apolipoprotein A-II (apoA-II) deposits as amyloid fibrils (AApoAII) and can be transmitted from one animal to another both by the feces and milk excreted by mice with amyloidosis. Thus, mouse AApoAII amyloidosis has been demonstrated to be a "transmissible disease". In this study, to further characterize the transmissibility of amyloidosis, AApoAII amyloid fibrils were injected into transgenic Apoa2(c)Tg(+/-) and normal R1.P1-Apoa2(c) mice to induce AApoAII systemic amyloidosis. Two months later, AApoAII amyloid deposits were found in the skeletal muscles of amyloid-affected mice, primarily in the blood vessels and in the interstitial tissues surrounding muscle fibers. When amyloid fibrils extracted from the skeletal muscles were subjected to Western blot analysis, apoA-II was detected. Amyloid fibril fractions isolated from the muscles not only demonstrated the structure of amyloid fibrils but could also induce amyloidosis in young mice depending on its fibril conformation. These findings present a possible pathogenesis of amyloidosis: transmission of amyloid fibril conformation through muscle, and shed new light on the etiology involved in amyloid disorders.