Nanobacteria and Psammoma Bodies: Ultrastructural Observations in a Case of Pathological Placental Calcification

Nanobacteria are controversial infectious agents with nanometric size, the capacity to nucleate hydroxyapatite and grow in culture, and present in human diseases associated with calcification and psammoma bodies. The authors report a case of pathological placental calcifications associated with nanobacteria. Electron microscopy and electron energy loss spectroscopy imaging were used to recognize 160-nm-sized calcium-free bodies mainly presenting as extracellular fibrillary tangles and 500-nm-sized calcified bodies; they encrusted the syncito-trophoblast basal membrane and aggregated into miniaturized psammoma bodies. Nanobacteria may be composed of a prionoid protein with self-assembling and self-propagating abilities whose growth is associated with the formation of psammoma bodies.     

Hypermineralized Whale Rostrum as the Exemplar for Bone Mineral

Abstract

Although bone is a nanocomposite of mineral and collagen, mineral has been the more elusive component of study. A standard for bone mineral is clearly needed. We hypothesized that the most natural, least-processed bone mineral could be retrieved from the most highly mineralized bone. We therefore studied the rostrum of the toothed whale Mesoplodon densirostris, which has the densest recognized bone. Essential to establishment of a standard for bone mineral is the documentation that the proposed tissue is bone-like in all properties except for its remarkably high concentration of mineral. Transmitted-light microscopy of unstained sections of rostral material shows normal bone morphology in osteon geometry, lacunae concentration, and vasculature development. Stained sections reveal extremely low density of thin collagen fibers in most of the bone, but enrichment of thicker collagen fibers around vascular holes and in a minority of osteons. Field-emission scanning electron microscopy shows the rostrum mostly consists of dense mineral prisms. Most rostral areas have the same chemical–structural features, i.e., Raman spectroscopically dominated by strong bands at ～962 Δcm^–1 and weak bands at ～2940 Δcm^–1. Spectral features indicate that the rostrum is composed mainly of the calcium phosphate mineral apatite and has only about 4 wt.% organic content. The degree of carbonate substitution (～8.5 wt.% carbonate) in the apatite is in the upper range found in most types of bone. We conclude that, despite its enamel-like extraordinarily high degree of mineralization, the rostrum is in all other features bone-like. Its mineral component is the long-sought uncontaminated, unaltered exemplar of bone mineral.     

Transmission Electron Microscopic Study of the Hemorrhagic Spots in Patients with Epidemic Hemorrhagic Fever in the Early Stage

Epidemic hemorrhagic fever (EHF) virus particles were found in the squamous epithelial cells and the capillary endothelial cells of the petechial spots located on the mucous membrane of the soft palate in 3 patients with severe early-stage EHF with transmission electron microscopy. The virus particles were round or oval in shape, about 100 nm in diameter, with a two-layer lipid envelope from which spikes were protruding. The nucleocapsid of the virus appeared to be hollow micro-filamentoid or dense granules. Meanwhile, budding virus particles with a diameter of 80 nm were found in the enlarged Golgi apparatus. The infected cells displayed an enlarged and proliferating Golgi apparatus. The morphological characteristics of the viron mentioned above coincided with those of the virus particles of the family Bunyaviridae. This study is the first to demonstrate that the squamous epithelial cells on the mucous membrane of the soft palate are the target cells of EHF infection and to provide subcellular morphological evidence of petechial hemorrhage at the soft palate.     

改性玻璃陶瓷在模拟体液中类骨磷灰石层形成的研究

研究用等离子体活化改性玻璃陶瓷(BGC)，并用模拟体液(SBF)中类骨磷灰石的形成、体外成骨细胞培养、SEM、XPS、XRD等对其进行表征。结果表明：与活化改性前相比较，等离子体活化改性后的BGC更有利于类骨磷灰石的形成，并能促进成骨细胞增殖。等离子体中丰富的高能、高活性的粒子轰击BGC，使其被刻蚀和粗化，增加了表面的溶解性和提供了更多的活性位点，易使局域的钙、磷离子浓度达到过饱和，更利于类骨磷灰石的成核和生长。表明等离子体改性提高了BGC的生物活性。     

粘蛋白在胆固醇结石和胆囊息肉胆囊组织中的表达和分泌差异

目的：研究粘蛋白（ mucin, Muc）在胆囊胆固醇结石和胆囊息内胆囊组织中的表达和分泌差异。方法纳入2010年6月至2015年6月在我院行胆囊切除术患者117为研究对象,其中单纯胆囊胆固醇结石患者78例,胆囊息肉患者39例,分析和比较两组患者胆囊组织中粘蛋白mRNA的相对表达水平,以及胆汁中粘蛋白的浓度,并进一步分析两者间的相关性。结果胆固醇结石患者胆囊组织中Muc1、 Muc2、Muc5AC和Muc5B mRNA的相对表达水平均明显高于胆囊息肉患者,其差异均具有统计学意义（P＜0．05）。而胆固醇结石患者胆囊组织中Muc3 mRNA的相对表达水平明显低于胆囊息肉患者,其差异具有统计学意义（P＜0．05）。另外,胆固醇结石患者胆汁中粘蛋白浓度明显高于胆囊息肉患者,其差异具有统计学意义（P＜0．05）。结论粘蛋白参与了胆囊胆固醇结石的形成和发展,可以作为胆囊胆固醇结石的治疗靶点。     

Gelsolin and Progression of Aortic Arch Calcification in Chronic Hemodialysis Patients

Background:Vascular calcification (VC) is a key process associated with cardiovascular mortality in dialysis patients. Gelsolin is an actin-binding protein that can modulate inflammation, correlated inversely with hemodialysis (HD) mortality and involved in bone calcification homeostasis. In this report, we aim to characterize progression in aortic arch calcification (AAC) and investigate its association with gelsolin.Methods: 184 HD patients were enrolled and their annual posterior-anterior chest X-ray films (CXR) in 2009 and 2013 were examined. The severity of AAC was classified as grade 0 to 3. Blood levels of gelsolin were measured by ELISA kits. Biographic and biochemical data at baseline were analyzed with status of AAC at baseline and changes after 4 years.Results: At baseline, 60% of the patients had detectable AAC on CXR. After 4 years, 77% had AAC. Patients with grade 1 and 2 AAC had increased risk of progression (Odds ratio [OR] 2~3, P=0.001) compared to those with grade 0 at baseline. Compared to those with no AAC, patients with AAC progression had older age, lower gelsolin, higher waist circumference and prevalence of vascular disease. Regression analysis confirmed baseline gelsolin (odds ratio 0.845, 95% confidence interval [0.734-0.974]) and waist circumference as the independent factors associated with AAC progression. Gelsolin is positively correlated with serum albumin and negatively with tumor necrosis factor-alpha.Conclusion: Our study demonstrated that HD patients with grades 1 or 2 baseline AAC are at increased risk of further progression compared to those with grade 0. We also found lower blood levels of gelsolin associated with progressive AAC. Further investigation into the mechanistic roles of gelsolin in vascular calcification may provide new understanding of this key process.     

模拟体液中生物大分子对类骨磷灰石矿化的影响

为考察体内生物大分子对羟基磷灰石（hydroxyapatite,HA）基底表面矿化物形成的影响,将牛血清白蛋白（bovine serum albumin,BSA）和硫酸软骨素（chondroitin sulfate,CS）大分子分别浸入模拟体液（SBF）中制备成2种矿化介质,再将HA浸入上述矿化介质中3d观察类骨磷灰石形成过程.结果 发现HA基底表面均沉积有Na＋和CO2-3取代的类骨磷灰石（Ca3.78Na0.02）（Ca5.22Na0.48）（CO3）1.5（OH）.BSA在2SBF中的存在促进了类骨磷灰石晶体在基材表面沉积,有利于其沿（300）晶面择优取向生长.CS对类骨磷灰石晶体的生长呈阻碍作用,获得的晶粒尺寸较小.模拟体液中BSA和CS大分子对类骨磷灰石晶体生长和形貌等均有一定的作用.     

Volumetric Analysis of Gallbladder in Extremely Premature Infants

Background

We hypothesized that gallbladder (GB) volume is affected by serial changes during the early infancy period in extremely premature infants.

在不同动物肌肉中磷酸钙陶瓷表面类骨磷灰石的形成研究

磷酸钙陶瓷材料植入动物体内后其表面类骨磷灰石层的形成对骨的形成有非常重要的作用，并被认为是骨诱导发生的先决条件。我们将相同大小的孔壁有微孔的多孔材料和致密磷酸钙陶瓷材料植入猪，狗、兔和鼠的背肌或腿肌内，研究陶瓷表面类骨磷灰石的形成，以了解类骨磷灰石层的形成与骨诱导的联系。结果表明：磷酸钙陶瓷材料植入动物的肌肉内14d后，狗，兔和鼠体内的多孔材料孔隙内表面（包括陶瓷表面较深孔隙）有一层类骨磷灰石层形成；植入猪体内的多孔材料内外表面都形成了一层类骨磷灰石，致密材料在几种动物体内都未观察到类骨磷灰石层形成，类骨磷灰石层形成的快慢次序与动物组织学观察到的在不同动物的肌内骨诱导性高低的次序不一致。证实了类骨磷灰石层的形成的确是骨诱导的先决条件。但还有其它因素影响骨诱导的发生。     

Limited Degradation of Gelatin Substrate by Cathepsin D

Cathepsin D has been demonstrated to degrade the protein backbone of proteinpolysaccharide. In this study we present evidence that cathepsin D has only a limited yet specific capacity to degrade gelatin. Similar to the effects of pepsin on collagen, cathepsin D degraded β chains of pure gelatin to α chains but showed no capacity to cleave at a site other than the N-terminal cross-link region. These data suggest that it is unlikely that cathepsin D has a major role in any phase of collagenolysis.     

Distribution and Ultrastructure of Afferent Fibers in the Parietal Peritoneum of the Rat

The distribution and the ultrastructure of afferent fibers innervating the parietal peritoneum in the rat was studied with immunohistochemistry using an antiserum against the neuronal marker protein gene product 9.5. The immunoreactive fibers were distributed throughout the peritoneum. They generally ran straight and parallel to the intercostal nerves running in the abdominal muscles underlying the peritoneum. They sometimes branched and terminated by forming club‐like endings. The number of nerve endings on the peritoneal surface was 3.25 ± 1.66 mm^?2. Electron microscopic observations revealed both unmyelinated and myelinated nerve fibers. The unmyelinated fibers were thin and about 1 μm in diameter. Their endings formed slight swellings located just inside the peritoneal cell layer. The myelinated fibers often formed a bundle that was composed of two or three nerve fibers. Each myelinated fiber kept in contact with a Shwann cell and projected toward the peritoneal cavity. Finally, they penetrated the peritoneal cell layer to reach the peritoneal cavity. These fibers then made contacts with the peritoneal cells and became free from the myelin sheath. The ending had a club‐like shape covered with collagen fibers, and contained many neurofilaments, a few mitochondria, but no synaptic vesicles. These results suggest that since the sensory endings are exposed at the peritoneal cavity, the sensory fibers are highly sensitive to somatic or nociceptive stimuli. Anat Rec, 2011. © 2011 Wiley‐Liss, Inc.     

Distribution of tracheal and laryngeal mucous glands in some rodents and the rabbit

We used scanning electron microscopy to count the number of mucous gland openings in the tracheae and lower portion of the larynges of the rat, guinea pig, hamster, mouse and rabbit. Cells of the airway surface epithelium were removed by protease digestion better to visualise the gland openings. The distribution of glands was further studied by conventional histology and by PAS/Alcian blue staining of whole mounts. In all rodent species, gland openings in the larynx occurred with a frequency of 1–2 per mm². Mice had no gland openings in their tracheae, and hamsters, only a handful. Rat tracheae contained 126±42 gland openings (± S.D. ; n = 6) at a frequency of ∼ 0.6 per mm² at the top of the trachea and ∼ 0.15 per mm² at the bottom. Guinea pig tracheae contained 153±90 gland openings (± S.D. ; n = 5), with 54% being in the top 40% of the trachea. In both rat and guinea pig, tracheal glands were found in the ventral aspect between the cartilaginous rings, and were absent from the dorsal membranous portion. Gland openings in most species were simple circles of ∼ 50 μm diameter. However, glands in the rat trachea generally opened obliquely into shallow (∼ 20 μm deep) oval troughs (∼ 150×75 μm), which had their long axes oriented from head to tail. In the rabbit, there was no evidence of tracheal or laryngeal glands histologically. However, the tracheal and laryngeal surfaces contained numerous pits (∼ 30 μm diameter) distributed evenly over and between cartilages at a frequency of ∼ 4 per mm². These may correspond to the 'nests' of goblet cells described by others.     

在动态模拟体液中致密CaP陶瓷表面形貌对类骨磷灰石层形成的影响研究

磷酸钙陶瓷植入体内后其表面类骨磷灰石层的形成是诱导成骨的先决条件。本实验在模拟体液 (Simu-lated body fluid,SBF)以人体骨骼肌组织的正常生理流率 (2 ml/ 10 0 m l· min)下 ,研究在动态 SBF中致密磷酸钙陶瓷表面形貌对类骨磷灰石层形成的影响。结果表明 :在生理流速条件下 ,材料的粗糙表面有利于类骨磷灰石层的形成 ,加大 SBF中 Ca2 +、HPO4 2 -离子浓度 ,类骨磷灰石层的形成速度加快。本研究进一步证实了材料的几何形貌对类骨磷灰石形成的影响 ,加深了对磷酸钙陶瓷在体内诱导成骨机理的理解     

原发性胆囊癌CT诊断与鉴别诊断

目的 探讨原发性胆囊癌的CT诊断与鉴别诊断要点。方法 回顾性分析23例原发性胆囊癌CT征象，并与手术和病理对照。结果 本组23例胆囊癌术前CT正确诊断18例，5例误诊，根据CT表现可分为：腔内型8例、厚壁型5例、肿块型10例；其中共8例并肝门和腹膜后淋巴结转移；3例肝内转移；病理结果：腺癌20例，鳞癌3例。结论 原发性胆囊癌CT的诊断价值较高，但仍容易误诊，早期的诊断仍有一定困难，掌握其CT征象，注意鉴别诊断可以提高本病的诊断水平。     

SLC20A2 Deficiency in Mice Leads to Elevated Phosphate Levels in Cerbrospinal Fluid and Glymphatic Pathway‐Associated Arteriolar Calcification,and Recapitulates Human Idiopathic Basal Ganglia Calcification

Idiopathic basal ganglia calcification is a brain calcification disorder that has been genetically linked to autosomal dominant mutations in the sodium‐dependent phosphate co‐transporter, SLC20A2. The mechanisms whereby deficiency of Slc20a2 leads to basal ganglion calcification are unknown. In the mouse brain, we found that Slc20a2 was expressed in tissues that produce and/or regulate cerebrospinal fluid, including choroid plexus, ependyma and arteriolar smooth muscle cells. Haploinsufficient Slc20a2 +/? mice developed age‐dependent basal ganglia calcification that formed in glymphatic pathway‐associated arterioles. Slc20a2 deficiency uncovered phosphate homeostasis dysregulation characterized by abnormally high cerebrospinal fluid phosphate levels and hydrocephalus, in addition to basal ganglia calcification. Slc20a2 siRNA knockdown in smooth muscle cells revealed increased susceptibility to high phosphate‐induced calcification. These data suggested that loss of Slc20a2 led to dysregulated phosphate homeostasis and enhanced susceptibility of arteriolar smooth muscle cells to elevated phosphate‐induced calcification. Together, dysregulated cerebrospinal fluid phosphate and enhanced smooth muscle cell susceptibility may predispose to glymphatic pathway‐associated arteriolar calcification.     

胆囊三维建模与运动功能分析

目的 考虑在健康和病理情况下的胆囊特性,研究胆囊壁生物力学和胆汁流体力学特性,以及胆囊运动功能与胆结石成因的关系。方法 利用CT影像数据构建胆囊三维结构模型,建立有限元模型模拟胆囊运动和胆汁流动过程,分析不同的流固耦合条件下胆囊的运动模式以及对胆结石形成的影响。结果 在相同边界条件下,结石性胆汁流速(1.1 cm/s)比正常胆汁流速(2.3 cm/s)低;胆囊运动功能异常与否不会改变胆汁的流型,而胆汁成分改变使得流型更加曲折。运动功能较弱的胆囊整体所受最大应力为1.079 kPa、最大形变为0.931 mm,均远小于健康胆囊相对应的最大应力(4.318 kPa)和最大形变(3.725 mm)。在受力方面,胆汁成分异常并未对胆囊造成太大影响。结论 胆囊运动功能和生物力学特性密切相关。胆囊受损或衰退后,其收缩量会大幅减少,胆汁流动不畅,更易导致结石产生。胆囊的三维模型和运动功能分析可为胆结石手术治疗提供必要的理论基础和技术手段。     

Oriented Growth of Octacalcium Phosphate on and inside the Collagenous Matrix in Vitro

To investigate the factors which regulate the growth of apatite in a collagenous matrix, a calcification experiment was carried out in a model system, where Ca²⁺ and PO₄^3- ions diffused into a slice of Achilles tendon from mutually opposite sides, at pH 6.5–7.0 and at 37°C for 1–3 days. Ribbon-like octacalcium phosphate (OCP) crystals (10–30 μm in length) grew on the Ca-side of the collagenous matrix, while small plate-like OCP crystals (<1 μm) grew inside the matrix. The major part of crystals grew with the c-axis parallel to the collagen fibers. Others grew with the c-axis parallel to the collagen bands or perpendicular to the fibrils. Crystals grew with orientation on collagen fibrils from the beginning. It was concluded that collagen fibrils and property of the collagenous matrix played a regulatory role in the deposition and growth of OCP in the collagenous matrix.     

50例胆囊癌黏液组化与免疫组化观察

目的：探讨黏液分泌变化和肿瘤相关抗原表达对胆囊癌早期诊断及其预后判断的价值。方法：采用黏液组化和免疫组化染色对40例胆囊癌,10例胆囊腺瘤,10例慢性胆囊炎的黏液分泌和CEA,CA50,E－cad及PCNA表达进行检测。结果：胆囊癌与胆囊炎和腺瘤相比,分泌硫酸黏液明显减少,唾酸黏液显著增多（P＜0．05）。胆囊癌CEA,CA50阳性表达率显著高于腺瘤和胆囊炎组（P＜0．05）,而E－cad阳性表达率明显降低,伴有转移者E－cad表达更低,胆囊癌PCNA LI高于胆囊炎和腺瘤组（P＜0．01）,CEA,PCNA过度表达者其3年生存率显著降低（P＜0．01）,E－cad过度表达者其3年生存率较阴性组明显增高（P＜0．05）。结论：黏液组化染色和CEA,CA50以及PCNA检测可能有助于胆囊腺瘤恶变和胆囊癌的早期诊断,且CEA PCNA和E－cad检测可作为胆囊癌预后判断的参考指标。     

甲状腺结节钙化在超声诊断甲状腺癌中的价值

目的 分析甲状腺结节钙化在超声诊断甲状腺癌中的价值。方法 选取2017年1月~2019年5月在我院诊治的94例甲状腺结节伴钙化患者临床资料,比较术前超声诊断钙化灶情况与临床病理诊断结果,分析甲状腺结节钙化与超声诊断甲状腺癌之间的相关性。结果 病理检查确诊良性结节56例,恶性结节38例;超声检查显示恶性结节患者的钙化结节单发76.31%、边界不清晰83.92%、钙化灶微钙化71.05%,均高于良性结节患者的44.64%、10.52%、28.57%,差异有统计学意义（P＜0.05）;恶性甲状腺肿瘤患者病理类型不同,其结节钙化情况也不同,甲状腺结节微钙化比均高于粗钙化比,差异有统计学意义（P＜0.05）。结论 甲状腺结节钙化可作为临床超声诊断与鉴别甲状腺癌的重要依据,结节单发、边界不清晰、钙化灶微钙化有助于甲状腺癌的鉴别诊断。