Bilateral medulloepithelioma of the ciliary body:a case report

Dear Editor,We present an exceptionally rare case about bilateral medulloepithelioma of the ciliary body.This case was approved by Ethics Committee of Capital Medical University.Written informed consent was obtained from the patients.Discovering leukocoria in both eyes and proptosis in right eye for 2mo by parents in a 4.5-month-old boy.He had undergone no treatment elsewhere except magnetic resonance imaging examination before coming to Beijing Tongren Hospital.

Clinical spectrum of primary ciliary dyskinesia in childhood

Although the triad of bronchiectasis, sinusitis and situs inversus was first described by Kartagener in 1933, the clinical spectrum of primary ciliary dyskinesia is still under investigation. Heterotaxy defects as well as upper and lower respiratory tract symptoms are the main manifestations in childhood. It is now recognized that situs inversus is encountered in only half of patients. The first lower respiratory symptoms may be present from infancy as neonatal respiratory distress. The most common lower airway manifestations are chronic wet cough, recurrent pneumonia and therapy resistant wheezing. Patients are at risk of developing bronchiectasis which may even be the presenting finding due to delayed diagnosis. Upper respiratory tract infections such as nasal congestion, nasal drainage and recurrent sinusitis as well as otologic manifestations such as otitis media or otorrhea with conductive hearing loss are also often encountered. It seems that the type of ciliary ultrastructure defects and the involved mutated genes are associated to some extent to the clinical profile. The disease, even in nowadays, is not recognized at an early age and the primary care clinician should have knowledge of its clinical spectrum in order to select appropriately the children who need further investigation for the diagnosis of this disorder.

Biomimetic Cilia Based on MEMS Technology

A review on the research of Micro Electromechanical Systems （MEMS） technology based biomimetic cilia is presented. Biomimetic cilia, enabled by the advancement of MEMS technology, have been under dynamic development for the past decade. After a brief description of the background of cilia and MEMS technology, different biomimetic cilia applications are reviewed. Biomimetic cilia micro-actuators, including micromachined polyimide bimorph biomimetic cilia micro-actuator, electro-statically actuated polymer biomimetic cilia micro-actuator, and magnetically actuated nanorod array biomimetic cilia micro-actuator, are presented. Subsequently micromachined underwater flow biomimetic cilia micro-sensor is studied, followed by acoustic flow micro-sensor. The fabrication of these MEMS-based biomimetic cilia devices, characterization of their physical properties, and the results of their application experiments are discussed.

Effect of Cilia Orientation in Metachronal Transport of Microparticles

A biomimetic approach is used to generate a directed transversal transportation of micron-sized particles in liquids based on the principle of cilia-type arrays in coordinated motion. Rows of flaps mimicking planar cilia are positioned off-centre along an array of cavities covered with membranes that support the flaps. These membranes are deflected from a concave to a convex shape and vice versa by pneumatic actuation applying positive and negative pressures (relative to the ambient) inside the cavities. As a result, the flap on top of the membrane tilts to the left or right within such a pressure cycle, performing a beat stroke. Since each cavity can be addressed in the device individually and in rapid succession, waves of coordinated flap motion can be run along the wall. Such metachronal waves are generated and transport of particles along the cilia surface is achieved in both symplectic and antiplectic direction. It is shown that the initial tilt of the flaps relative to the wall-normal determines the direction of transport.

Rheological fluid motion in tube by metachronal waves of cilia

This paper presents a theoretical study of a non-linear rheological fluid transport in an axisymmetric tube by cilia. An attempt has been made to explain the role of cilia motion in the transport of fluid through the ductus efferent of the male reproductive tract. The Ostwald-de Waele power-law viscous fluid is considered to represent the rheological fluid. We analyze pumping by means of a sequence of cilia beats from rowto-row of cilia in a given row of cells and from one row of cells to the next(metachronal wave movement). For this purpose, we consider the conditions that the corresponding Reynolds number is small enough for inertial effects to be negligible, and the wavelengthto-diameter ratio is large enough so that the pressure can be considered uniform over the cross section. Analyses and computations of the fluid motion reveal that the time-average flow rate depends on ε, a non-dimensional measure involving the mean radius a of the tube and the cilia length. Thus, the flow rate significantly varies with the cilia length.Moreover, the flow rate has been reported to be close to the estimated value 6 × 10ml/h for human efferent ducts if ε is near 0.4. The estimated value was suggested by Lardner and Shack(Lardner, T. J. and Shack, W. J. Cilia transport. Bulletin of Mathematical Biology, 34, 325–335(1972)) for human based on the experimental observations of flow rates in efferent ducts of other animals, e.g., rat, ram, and bull. In addition, the nature of the rheological fluid, i.e., the value of the fluid index n strongly influences various flow-governed characteristics. An interesting feature of this paper is that the pumping improves the thickening behavior for small values of ε or in free pumping(?P = 0) and pumping(?P > 0) regions.

Cilia containing 9+2 structures grown from immortalized cells

Cilia depend on their highly differentiated structure, a 9 ＋ 2 arrangement, to remove particles from the lung and to transport reproductive cells. Immortalized cells could potentially be of great use in cilia research. Immortalization of cells with cilia structure containing the 9 ＋ 2 arrangement might be able to generate cell lines with such cilia structure. How- ever, whether immortalized cells can retain such a highly differentiated structure remains unclear. Here we demonstrate that （1） using Ela gene transfection, tracheal cells are immortalized; （2） interestingly, in a gel culture the immortalized cells form spherical aggregations within which a lumen is developed; and （3） surprisingly, inside the aggregation, cilia containing a 9 ＋ 2 arrangement grow from the cell＇s apical pole and protrude into the lumen. These results may influence future research in many areas such as understanding the mechanisms of cilia differentiation, cilia generation in other existing cell lines, cilia disorders, generation of other highly differentiated structures besides cilia using the gel culture, immortalization of other ciliated cells with the Ela gene, development of cilia motile function, and establishment of a research model to provide uniform ciliated cells.

Case report: Metastatic adenocarcinoma of the lung with filiform “ciliated” tumor cells in the cerebrospinal fluid

Occurrence of cilia on the cell surface is usually considered evidence of a non-neoplastic process. Rarely, ciliated malignant tumors cells have been reported in pleural and ascitic fluids from ovarian and endometrial adenocarcinomas, though not from the cerebrospinal fluid (CSF) in association with pulmonary adenocarcinoma. We now present a case report of a patient who initially presented with neurologic symptoms. A CSF specimen revealed atypical cells with peripheral cilia and some cytoplasmic pigment. Cytologic, morphologic, and immunohistochemical analyses established this to be a metastatic adenocarcinoma of pulmonary origin with metastases to the meninges, pelvis, and the vertebrae. Cilia are generally the antithesis of malignancy as their presence is considered to be evidence of proper regulation of the cell cycle within a well-differentiated cell. This case offers the first described example of malignant ciliated cells derived from a metastatic lung adenocarcinoma. An awareness of this unusual presentation should aid in diagnosis and management when similar situations are prospectively encountered.

De novo identification of mammalian ciliary motility proteins using cryo-EM

Dynein-decorated doublet microtubules(DMTs)are critical components of the oscillatory molecular machine of cilia,the axoneme,and have luminal surfaces patterned periodically by microtubule inner proteins(MIPs).Here we present an atomic model of the 48-nm repeat of a mammalian DMT,derived from a cryoelectron microscopy(cryo-EM)map of the complex isolated from bovine respiratory cilia.

Diagnosis and treatment of male infertility associated with immotile cilia syndrome:a report of 6 cases

Objective:To investigate the clinical characteristics,diagnosis and treatment of male infertility associated with immotile cilia syndrome(ICS). Methods:The clinical data of six cases of male infertility associated with ICS were reviewed retrospectively. Results:The clinical features in all cases included chronic or recurrent infections of the upper and lower airways and male infertility.Sinusitis,bronchitis and bronchiectasis were found in all cases and situs inversus totalis found in two cases.Sex hormone levels,chromosome karyotype and Y chromosome microdeletion(AZF) in all cases were normal.All cases were diagnosed as severe asthenospermia by routine semen analysis.Five cases had no motile spermatozoa in semen,while there were less than 0.2%of motile sperm in one case.The total sperm count and sperm viability were normal in four cases and there were very few immotile sperm in two cases.Transmission electron microscopic examination of sperm flagellum revealed disarrangement or a partial absence of 9+2 microtubules and/or an absence of the dynein arms in six cases.The bronchial cilia in one case showed to be devoid of inner dynein arms.Five cases underwent six intracytoplasmic sperm injection(ICSI) cycles and the rates of fertilization,embryo cleavage and good quality embryos were 50.0%,69.2%and 55.6%,respectively.Two clinical pregnancies and one chemical pregnancy were achieved,with one birth of a healthy baby boy. Conclusions:The ultrastructural defect of cilia or flagellum is the most important diagnostic criteria of ICS. ICSI is an effective treatment for male infertility associated with ICS.

The role of primary cilia in mechanical transmission of osteocyte based on a 3D finite element model

The primary cilium,as a mechanical receptor of osteocytes,participates in the regulation of osteocyte mechanosensitivity.However,how the length of osteocyte primary cilia changes with fluid shear stress(FSS)are unclear,and how the mechanical transmission within osteocytes altered by primary cilia is not well understood yet.Therefore,the ciliary length changes of osteocyte under 15dyn/cm2 of FSS were experimentally detected,and then 3D finite element models of osteocyte primary cilia containing the basal body and axoneme were built.The results showed that(1)The ciliary length of the CON group,FSS 1h,and FSS 6h were 3.71±1.34μm,3.79±1.04μm,and 1.24±0.73μm respectively,indicating the different durations of FSS might lead to the adaptive changes of cilium length.The calculations showed(2)when the ciliary length became shorter with the ciliary angle stayed the same,the deformation and stress of the cell membrane and membrane skeleton was increased.However,the deformation and stress of the cilia membrane,basal body,the rotation angles of basal body were decreased,and those of cytoplasm,cytoskeleton,actin cortex and nucleus were also decreased;(3)With the decrease of the ciliary angle,the deformation and stress of the cilia membrane,basal body,as well as the rotation angles of basal body were increased.Those of the cytoplasm,cytoskeleton,actin cortex,and nucleus were also increased except the cell membrane and membrane skeleton.The calculation results suggested the length and angle of the primary cilia,the deformation and stress of intracellular structures in osteocyte were altered with ciliary basal body,indicated the connection between the basal body and cytoskeleton may be a key factor that affected the mechanical transport in osteocytes across the cell membrane.This finally promoted the adaptive change of ciliary length under FSS.

Dynein axonemal heavy chain 10 deficiency causes primary ciliary dyskinesia in humans and mice

Primary ciliary dyskinesia(PCD)is a congenital,motile ciliopathy with pleiotropic symptoms.Although nearly 50 causative genes have been identified,they only account for approximately 70%of definitive PCD cases.Dynein axonemal heavy chain 10(DNAH10)encodes a subunit of the inner arm dynein heavy chain in motile cilia and sperm flagella.Based on the common axoneme structure of motile cilia and sperm flagella,DNAH10 variants are likely to cause PCD.Using exome sequencing,we identified a novel DNAH10 homozygous variant(c.589C>T,p.R197W)in a patient with PCD from a consanguineous family.The patient manifested sinusitis,bronchiectasis,situs inversus,and asthenoteratozoospermia.Immunostaining analysis showed the absence of DNAH10 and DNALI1 in the respiratory cilia,and transmission electron microscopy revealed strikingly disordered axoneme 9+2 architecture and inner dynein arm defects in the respiratory cilia and sperm flagella.Subsequently,animal models of Dnah10-knockin mice harboring missense variants and Dnah10-knockout mice recapitulated the phenotypes of PCD,including chronic respiratory infection,male infertility,and hydrocephalus.To the best of our knowledge,this study is the first to report DNAH10 deficiency related to PCD in human and mouse models,which suggests that DNAH10 recessive mutation is causative of PCD.

Lack of CFAP54 causes primary ciliary dyskinesia in a mouse model and human patients

Primary ciliary dyskinesia(PCD)is a highly heterogeneous recessive inherited disorder.FAP54,the homolog of CFAP54 in Chlamydomonas reinhardtii,was previously demonstrated as the C1d projection of the central microtubule apparatus of flagella.A Cfap54 knockout mouse model was then reported to have PCD-relevant phenotypes.Through whole-exome sequencing,compound heterozygous variants c.2649_2657delinC(p.E883Dfs*47)and c.7312_7313insCGCAGGCTGAATTCTTGG(p.T2438delinsTQAEFLA)in a new suspected PCD-relevant gene,CFAP54,were identified in an individual with PCD.Two missense variants,c.4112A>C(p.E1371A)and c.6559C>T(p.P2187S),in CFAP54 were detected in another unrelated patient.In this study,a minigene assay was conducted on the frameshift mutation showing a reduction in mRNA expression.In addition,a CFAP54 in-frame variant knock-in mouse model was established,which recapitulated the typical symptoms of PCD,including hydrocephalus,infertility,and mucus accumulation in nasal sinuses.Correspondingly,two missense variants were deleterious,with a dramatic reduction in mRNA abundance from bronchial tissue and sperm.The identification of PCD-causing variants of CFAP54 in two unrelated patients with PCD for the first time provides strong supportive evidence that CFAP54 is a new PCD-causing gene.This study further helps expand the disease-associated gene spectrum and improve genetic testing for PCD diagnosis in the future.

Ccdc57 is required for straightening the body axis by regulating ciliary motility in the brain ventricle of zebrafish

Recently,cilia defects have been proposed to contribute to scoliosis.Here,we demonstrate that coiled-coil domain-containing 57(Ccdc57)plays an essential role in straightening the body axis of zebrafish by regulating ciliary beating in the brain ventricle(BV).Zygotic ccdc57(Zccdc57)mutant zebrafish developes scoliosis without significant changes in their bone density and calcification,and the maternal-zygotic ccdc57(MZccdc57)mutant embryos display curved bodies since the long-pec stage.The expression of ccdc57 is enriched in ciliated tissues and immunofluorescence analysis reveals colocalization of Ccdc57-HA with acetylated a-tubulin,implicating it in having a role in ciliary function.Further examination reveals that it is the coordinated cilia beating of multiple cilia bundles(MCB)in the MZccdc57 mutant embryos that is affected at 48 hours post fertilization,when the compromised cerebrospinal fluid flow and curved body axis have already occurred.Either ccdc57 m RNA injection or epinephrine treatment reverses the spinal curvature in MZccdc57 mutant larvae from ventrally curly to straight or even dorsally curly and significantly upregulates urotensin signaling.This study reveals the role of ccdc57 in maintaining coordinated cilia beating of MCB in the BV.

The actin-bundling protein Fascin-1 modulates ciliary signalling

Primary cilia are microtubule-based cell organelles important for cellular communication. Since they are involved in the regulation of numerous signalling pathways, defects in cilia development or function are associated with genetic disorders, collectively called ciliopathies. Besides their ciliary functions, recent research has shown that several ciliary proteins are involved in the coordination of the actin cytoskeleton. Although ciliary and actin phenotypes are related, the exact nature of their interconnection remains incompletely understood. Here, we show that the protein BBS6, associated with the ciliopathy Bardet–Biedl syndrome, cooperates with the actin-bundling protein Fascin-1 in regulating filopodia and ciliary signalling. We found that loss of Bbs6 affects filopodia length potentially via attenuated interaction with Fascin-1. Conversely, loss of Fascin-1 leads to a ciliary phenotype, subsequently affecting ciliary Wnt signalling, possibly in collaboration with BBS6. Our data shed light on how ciliary proteins are involved in actin regulations and provide new insight into the involvement of the actin regulator Fascin-1 in ciliogenesis and cilia-associated signalling. Advancing our knowledge of the complex regulations between primary cilia and actin dynamics is important to understand the pathogenic consequences of ciliopathies.

IFT52 plays an essential role in sensory cilia formation and neuronal sensory function in Drosophila

Cilia are microtubule-based,hair-like organelles involved in sensory function or motility,playing critical roles in many physiological processes such as reproduction,or-gan development,and sensory perception.In insects,cilia are restricted to certain sensory neurons and sperms,being important for chemical and mechanical sensing,and fertility.Although great progress has been made regarding the mechanism of cilia assembly,the formation of insect cilia remains poorly understand,even in the insect model organism Drosophila.Intraflagellar transport(IFT)is a cilia-specific complex that traffics protein cargos bidirectionally along the ciliary axoneme and is essential for most cilia.Here we investigated the role of IFT52,a core component of IFT-B,in cilia/flagellar formation in Drosophila.We show that Drosophila IFT52 is distributed along the sensory neuronal cilia,and is essential for sensory cilia formation.Deletion of Ift52 results in severe defects in cilia-related sensory behaviors.It should be noted that IFT52 is not detected in sper-matocyte cilia or sperm flagella of Drosophila.Accordingly,ift52 mutants can produce sperms with normal motility,supporting a dispensable role of IFT in Drosophila sperm flagella formation.Altogether,IFT52 is a conserved protein essential for sensory cilia formation and sensory neuronal function in insects.

用于龙舟训练的仿生柔性触觉传感器

为了提高龙舟比赛运动员的训练效果,开发了可用于身体运动识别及压力监测的仿生纤毛柔性触觉传感器。柔性触觉传感器采用了经典的基底-传感层-封装层式“三明治”结构,柔性硅胶基底采用旋涂法制备,传感层为利用Langmuir⁃Blodgett方法得到的大面积碳纳米管传感薄膜,纤毛栽种于传感层,最后封装得到具有仿生纤毛的触觉传感器。由传感器的SEM照片可知,层与层之间的结合特性较好,采用自制的往复测试平台,对传感器进行了测试。传感器耐用性最高可达5500次,最大阻值差可达300%,对于不同方向力的刺激有识别特性,可清晰识别人体的脉搏信号,对未来龙舟训练智能装备开发中的动作识别、力量监测等功能具有较大现实意义。

Ricolinostat驱动初级纤毛的解聚降低了X线损伤的心肌纤维化

目的 评价Ricolinostat对X线辐射诱导心肌纤维化和心肌成纤维细胞初级纤毛损伤的影响。方法 8 Gy剂量单次全身照射Wistar大鼠,腹腔注射Ricolinostat, ELISA测定血清肌钙蛋白T(troponinT,TnT)和脑钠肽(brain natriuretic peptide, BNP)水平;HE和Masson染色检测心肌组织纤维化程度,免疫荧光检测心肌组织和心肌成纤维细胞初级纤毛及Ⅰ型胶原含量;qRT-PCR和Western blot检测心肌组织和心肌成纤维细胞组蛋白去乙酰化酶6(histone deacetylase 6,HDAC6)的基因和蛋白水平。结果 与对照组相比,X线辐射增加大鼠心肌组织胶原紊乱和Ⅰ型胶原含量,促进心肌成纤维细胞增殖,上调了心肌成纤维细胞中HDAC6的表达(P> 0.05),增加心肌组织和心肌成纤维细胞初级纤毛的形成。经过Ricolinostat治疗后,心肌组织和心肌成纤维细胞中HDAC6的表达水平和初级纤毛形成降低(P<0.05)。胶原纤维排列稍显整齐,胶原体积分数降低,血清中TnT(P<0.01)和BNP水平降低。结论 Ricolinostat能够驱动初级纤毛的解聚,降低了X线损伤诱导的心肌纤维化,具有治疗放射性心肌纤维化的潜在价值。

初级纤毛/鞭毛转运系统介导力学反应性信号通路促骨髓基质干细胞成骨分化

背景:目前已证实力学刺激可以促进骨髓基质干细胞成骨分化,但其机制未完全明了。初级纤毛是重要的力学感受器并调控TGF-β1/BMP-2/SMAD等多种信号通路,很可能是骨髓基质干细胞力学调控的重要靶点。目的:探讨流体剪切力对骨髓基质干细胞成骨分化的影响及机制。方法:将大鼠骨髓基质干细胞分为对照组、力学刺激组(通过摇床施加流体剪切力学干预)、力学刺激+IFT88沉默组(力学刺激+使用siRNA沉默IFT88表达),干预24 h后,采用qRT-PCR检测转化生长因子β1、骨形成蛋白2的表达、Western blot检测磷酸化SMAD2/3蛋白的表达,初级纤毛免疫荧光染色及形态学分析。结果与结论:剪切力刺激可促进骨髓基质干细胞的初级纤毛表达,转化生长因子β1及骨形成蛋白2基因转录激活,提高磷酸化SMAD2/3蛋白表达。siRNA干扰初级纤毛生成后,这一力学反应效应明显减低。骨髓基质干细胞的初级纤毛面积改变比值与转化生长因子β1及骨形成蛋白2基因转录增高比例具有Spearman相关性。结果表明:初级纤毛/鞭毛转运系统介导了流体剪切力反应性的TGF-β1/BMP-2/SMAD信号通路激活,促进骨髓基质干细胞成骨分化。

斑马鱼模型在特发性脊柱侧凸研究中的应用

特发性脊柱侧凸(IS)指原因不明的脊柱发育异常,给患者带来身体和经济的双重负担。然而,目前IS的致病机制尚未完全明确,疾病的治疗通常以大创口的外科手术干预为主。为了探索IS的病因,并为新的诊疗方法提供理论基础,采用各种模式动物进行实验尤为必要。作为一种新兴的模式动物,斑马鱼繁殖周期短、培育成本低,且具备体外受精、体外发育和胚胎透明等特点,因此其遗传编辑和观测均较传统模式动物更容易。目前斑马鱼模型已成为研究IS的重要工具,其在揭示IS的分子机制和潜在治疗靶点方面展现出巨大潜力。

Spag6调控运动纤毛结构功能缓解大鼠缺血性脑水肿损伤

目的 研究精子相关抗原6(sperm-associated antigen 6,Spag6)对缺血性脑水肿介导的突触损伤的影响。方法 将90只SD大鼠随机分为3组:假手术组、模型组和Spag6组。利用慢病毒感染上调大鼠脑内Spag6蛋白表达后,用大脑中动脉栓塞法构建缺血性脑水肿模型。利用Longa评分进行神经功能评价;2,3,5-氯化三苯基四氮唑(TTC)染色评估脑梗死体积;苏木精-伊红(HE)染色法、扫描电镜观察运动纤毛结构活性;透射电镜观察神经细胞超微结构;Western blot检测Bcl-2、Bax、Survivin、GluN1、GluN2B和GluA2蛋白表达水平。结果 模型组大鼠大脑不同区域神经细胞和组织损伤,Spag6表达异常变化,突触结构功能改变;第三脑室运动纤毛活性紊乱,发生缺血性脑水肿现象。上调Spag6表达后,脑水肿现象缓解,运动纤毛活性恢复;缺血区脑梗死、神经细胞损伤及突触结构异常改变被逆转;Bcl-2、Survivin、GluN1、GluN2B、GluA2的蛋白表达显著升高(P<0.01或P<0.05),Bax表达量均降低(P<0.01)。结论 Spag6通过恢复脑室运动纤毛的活性状态,增强突触的功能,进而减轻缺血性脑水肿介导的神经损伤。