Downregulation of HDAC1 suppresses media degeneration by inhibiting the migration and phenotypic switch of aortic vascular smooth muscle cells in aortic dissection

Although much progress has been made in the diagnosis and treatment of thoracic aortic dissection (TAD), the overall morbidity and mortality rates of TAD are still high. Therefore, the molecular pathogenesis and etiology of TAD need to be elucidated. In this study, we found that histone deacetylase 1 (HDAC1) expression is dramatically higher in the aortic wall of patients with TAD (than that in a normal group) and negatively correlates with the levels of the vascular smooth muscle cell (SMC) contractile-phenotype markers. Knockdown of HDAC1 upregulated both smooth muscle 22 α (SM22α) and α-smooth muscle actin (α-SMA) in platelet-derived growth factor (PDGF)-BB-treated and -untreated SMCs. In addition, the knockdown of HDAC1 markedly decreased SMC viability and migration in contrast to the control group under the conditions of quiescence and PDGF-BB treatment. We also showed that the expression of polycystic kidney disease 1 (PKD1) is decreased in the aortic wall of patients with TAD and negatively correlates with HDAC1 expression. Overexpressed PKD1 obviously increased SM22α and α-SMA expression and reduced the viability and migration of SMCs, but these effects were attenuated by HDAC1. Furthermore, we demonstrated that HDAC1 serves as an important modulator of the migration and phenotypic switch of SMCs by suppressing the PKD1– mammalian target of the rapamycin signaling pathway. HDAC1 downregulation inhibited media degeneration and attenuated the loss of elastic–fiber integrity in a mouse model of TAD. Our results suggest that HDAC1 might be a new target for the treatment of a macrovascular disease such as TAD.     

小切开EST联合EPBD与单纯EST对85岁以上老年胆总管结石患者疗效的比较

目的:对比内镜下十二指肠乳头括约肌(expressed sequence tags,EST)小切开术联合内镜下十二指肠乳头括约肌扩张术(endoscopic papillary balloon dilation,EPBD)与单纯EST对85岁以上老年胆总管结石患者的疗效。方法:选择我院于2014年1月～2020年2月收治的85岁以上老年胆总管结石患者150例,根据入院顺序随机分成两组,每组各75例,给予对照组单纯小切开EST术治疗,给予研究组小切开EST+EPBD术治疗。对比两组的一次取石成功率、机械碎石、结石复发率等指标;术中操作时间、术中出血量、住院天数、术后排便天数等临床指标;术后胆道感染、急性胰腺炎、高淀粉酶血症、术后腹痛等并发症的总发生率。结果:研究组一次取石成功率显著高于对照组,机械碎石、结石复发率均显著低于对照组(P0.05);研究组的术中操作时间、术中出血量、住院天数、术后排便天数均显著低于对照组(P0.05);研究组术后胆道感染、急性胰腺炎、高淀粉酶血症、术后腹痛、术后迟发性出血等并发症的总发生率为9.33%(7/75),显著低于对照组37.33%(28/75),差异具有统计学意义(P0.05)。结论:小切开EST联合EPBD对85岁以上老年胆总管结石患者的疗效显著,该方法可有改善患者临床指标,降低术后并发症发生率,值得推荐至临床广泛应用。     

洗胃法与剖胃法在四种蛙食性分析中的对比

在动物的食性研究中,剖胃法因要杀死动物,会产生一系列的生态学影响和伦理学问题,在国际上招致了越来越多的反对,而在我国仍广泛使用。尽管洗胃法已用于食性研究,它的效果却从来没有研究过。我们应用洗胃法研究了浙江宁波郭巨镇四种蛙(黑斑蛙、泽蛙、金线蛙和中华蟾蜍)的食性,并与剖胃法所获得的食性结果进行比较。结果显示:尽管洗胃法在四种蛙胃内食物量的总洗出率(在94.50%以上)和食物个体数的洗出率上(均大于93.46%)略低于剖胃法,但胃内未洗出的与洗出的平均食物个体大小间无差异(所有P>0.3061),两种方法获得的四种蛙的平均食物量和食物个体数也无差异(对食物量所有P>0.8680,对食物个体数所有P>0.7923)。结果表明,洗胃法是比较精确的,在动物的食性研究中,应广泛地推广洗胃法,摒弃剖胃法     

ERCP术治疗胆总管结石的有效性及安全性分析

目的:分析经内镜逆行胰胆管造影术(ERCP)术治疗胆总管结石的有效性及安全性。方法:选择2017年1月～2018年8月在我院接受择期ERCP治疗的164例胆总管结石患者为研究对象,分析患者的手术情况、取石效果、手术前后胃肠疾病生活质量指数(GIQLI)量表评分和并发症的发生情况。结果:胆总管结石患者手术时间为(37.90±4.21)min、术中出血量(10.86±1.29)mL、术后通气时间(4.38±0.65)d、切口疼痛时间(1.02±0.12)d、住院时间(8.62±0.96)d、手术成功率为97.56%(160/164)、一次取净结石率为95.73%(157/164)、二次取净结石率为1.82%(3/164)。术后,胆总管结石患者GIQLI评分均显著高于术前(P0.05)。胆总管结石患者术后发生胰腺炎5例、胆管炎1例、出血6例、高淀粉酶血症4例。结论:ERCP术是胆总管结石患者的有效治疗手段,但需积极预防并处理相关并发症。     

急性Stanford A型主动脉夹层患者的临床特征及术后院内死亡危险因素分析

目的:研究急性Stanford A型主动脉夹层(AAAD)患者的临床特征,并分析影响其术后院内死亡的危险因素。方法:选取2015年6月～2018年4月河北医科大学第四医院心脏血管外科收治的279例AAAD患者为研究对象,收集患者的基本信息、临床资料,分析AAAD患者的临床特征。所有患者均行手术治疗,按照术后院内的死亡情况将患者分为死亡组和存活组,收集两组患者手术相关信息,采用多因素logistic回归分析对患者术后死亡的危险因素进行分析。结果:279例AAAD患者中,男女比例为2.88:1;患者平均年龄为(52.11±8.91)岁;合并高血压者占66.67%,合并冠心病者占11.47%;患者以胸痛或胸背痛为主要症状(占69.89%);并发症中以心包积液(占43.37%)和主动脉瓣反流(占18.64%)的发生率最高;其平均收缩压、白细胞计数、D-二聚体(DDi)水平均高于正常值上限;心、肾功能不全的发生率分别为23.30%和15.77%;院内死亡率为17.92%。单因素分析结果显示,死亡组和存活组AAAD患者在年龄、是否伴有肾功能不全、是否伴有心功能不全、DDi水平、体外循环时间、手术时间、输血量和术后是否开胸止血方面比较,均有统计学差异(P0.05)。多因素logistic回归分析结果显示,年龄65岁、伴有肾功能不全、体外循环时间≥270 min和术后开胸止血是AAAD患者术后死亡的独立危险因素(P0.05)。结论:AAAD患者发病年龄呈年轻化,多数合并有基础疾病,疼痛为其主要症状,且该病患者并发症多、术后死亡率较高。年龄65岁、伴有肾功能不全、体外循环时间≥270min和术后开胸止血均可显著增加AAAD患者术后死亡风险。     

The Swimmeret System of Crayfish: A Practical Guide for the Dissection of the Nerve Cord and Extracellular Recordings of the Motor Pattern

Here we demonstrate the dissection of the crayfish abdominal nerve cord. The preparation comprises the last two thoracic ganglia (T4, T5) and the chain of abdominal ganglia (A1 to A6). This chain of ganglia includes the part of the central nervous system (CNS) that drives coordinated locomotion of the pleopods (swimmerets): the swimmeret system. It is known for over five decades that in crayfish each swimmeret is driven by its own independent pattern generating kernel that generates rhythmic alternating activity ^1-3. The motor neurons innervating the musculature of each swimmeret comprise two anatomically and functionally distinct populations ⁴. One is responsible for the retraction (power stroke, PS) of the swimmeret. The other drives the protraction (return stroke, RS) of the swimmeret. Motor neurons of the swimmeret system are able to produce spontaneously a fictive motor pattern, which is identical to the pattern recorded in vivo ¹.The aim of this report is to introduce an interesting and convenient model system for studying rhythm generating networks and coordination of independent microcircuits for students’ practical laboratory courses. The protocol provided includes step-by-step instructions for the dissection of the crayfish’s abdominal nerve cord, pinning of the isolated chain of ganglia, desheathing the ganglia and recording the swimmerets fictive motor pattern extracellularly from the isolated nervous system.Additionally, we can monitor the activity of swimmeret neurons recorded intracellularly from dendrites. Here we also describe briefly these techniques and provide some examples. Furthermore, the morphology of swimmeret neurons can be assessed using various staining techniques. Here we provide examples of intracellular (by iontophoresis) dye filled neurons and backfills of pools of swimmeret motor neurons. In our lab we use this preparation to study basic functions of fictive locomotion, the effect of sensory feedback on the activity of the CNS, and coordination between microcircuits on a cellular level.     

Saliva,Salivary Gland,and Hemolymph Collection from Ixodes scapularis Ticks

Ticks are found worldwide and afflict humans with many tick-borne illnesses. Ticks are vectors for pathogens that cause Lyme disease and tick-borne relapsing fever (Borrelia spp.), Rocky Mountain Spotted fever (Rickettsia rickettsii), ehrlichiosis (Ehrlichia chaffeensis and E. equi), anaplasmosis (Anaplasma phagocytophilum), encephalitis (tick-borne encephalitis virus), babesiosis (Babesia spp.), Colorado tick fever (Coltivirus), and tularemia (Francisella tularensis) ^1-8. To be properly transmitted into the host these infectious agents differentially regulate gene expression, interact with tick proteins, and migrate through the tick ^3,9-13. For example, the Lyme disease agent, Borrelia burgdorferi, adapts through differential gene expression to the feast and famine stages of the tick''s enzootic cycle ^14,15. Furthermore, as an Ixodes tick consumes a bloodmeal Borrelia replicate and migrate from the midgut into the hemocoel, where they travel to the salivary glands and are transmitted into the host with the expelled saliva ^9,16-19.As a tick feeds the host typically responds with a strong hemostatic and innate immune response ^11,13,20-22. Despite these host responses, I. scapularis can feed for several days because tick saliva contains proteins that are immunomodulatory, lytic agents, anticoagulants, and fibrinolysins to aid the tick feeding ^{3,11,20,21,23}. The immunomodulatory activities possessed by tick saliva or salivary gland extract (SGE) facilitate transmission, proliferation, and dissemination of numerous tick-borne pathogens ^3,20,24-27. To further understand how tick-borne infectious agents cause disease it is essential to dissect actively feeding ticks and collect tick saliva. This video protocol demonstrates dissection techniques for the collection of hemolymph and the removal of salivary glands from actively feeding I. scapularis nymphs after 48 and 72 hours post mouse placement. We also demonstrate saliva collection from an adult female I. scapularis tick.     

Dissection,Culture, and Analysis of Xenopus laevis Embryonic Retinal Tissue

The process by which the anterior region of the neural plate gives rise to the vertebrate retina continues to be a major focus of both clinical and basic research. In addition to the obvious medical relevance for understanding and treating retinal disease, the development of the vertebrate retina continues to serve as an important and elegant model system for understanding neuronal cell type determination and differentiation^1-16. The neural retina consists of six discrete cell types (ganglion, amacrine, horizontal, photoreceptors, bipolar cells, and Müller glial cells) arranged in stereotypical layers, a pattern that is largely conserved among all vertebrates ^12,14-18.While studying the retina in the intact developing embryo is clearly required for understanding how this complex organ develops from a protrusion of the forebrain into a layered structure, there are many questions that benefit from employing approaches using primary cell culture of presumptive retinal cells ^7,19-23. For example, analyzing cells from tissues removed and dissociated at different stages allows one to discern the state of specification of individual cells at different developmental stages, that is, the fate of the cells in the absence of interactions with neighboring tissues ^{8,19-22,24-33}. Primary cell culture also allows the investigator to treat the culture with specific reagents and analyze the results on a single cell level ^{5,8,21,24,27-30,33-39}. Xenopus laevis, a classic model system for the study of early neural development ^{19,27,29,31-32,40-42}, serves as a particularly suitable system for retinal primary cell culture ^10,38,43-45.Presumptive retinal tissue is accessible from the earliest stages of development, immediately following neural induction ^25,38,43. In addition, given that each cell in the embryo contains a supply of yolk, retinal cells can be cultured in a very simple defined media consisting of a buffered salt solution, thus removing the confounding effects of incubation or other sera-based products ^10,24,44-45.However, the isolation of the retinal tissue from surrounding tissues and the subsequent processing is challenging. Here, we present a method for the dissection and dissociation of retinal cells in Xenopus laevis that will be used to prepare primary cell cultures that will, in turn, be analyzed for calcium activity and gene expression at the resolution of single cells. While the topic presented in this paper is the analysis of spontaneous calcium transients, the technique is broadly applicable to a wide array of research questions and approaches (Figure 1).     

副沙鳅属系统发育分析

以沙鳅属Botia为外类群,共发现了副沙鳅属Parabotia 35个有系统发育意义的外部与骨骼形态特征,并由此重建副沙鳅属系统发育关系为:漓江副沙鳅 (双斑副沙鳅 (花斑副沙鳅 (武昌副沙鳅 点面副沙鳅)))或漓江副沙鳅 (双斑副沙鳅 (武昌副沙鳅 (点面副沙鳅 花斑副沙鳅))).具有囟门;后翼骨外侧面为不规则的梯形;吻骨侧面观呈一长方形;无下舌软骨;具有3对咽鳃骨;中喙骨弯曲较大,近直角;上吻皮中间有切刻状缺口,左右不连续;以及颐部无纽状突起等构成了副沙鳅属的共同离征.