改良革兰染色法在显示石蜡切片真菌中的应用

目的建立真菌的病理切片改良革兰染色法。方法选取确诊真菌感染的组织蜡块,切若干空白片,通过苏木素-伊红染色（HE）、高碘酸-无色品红染色（PAS）、六胺银染色（GMS）、改良革兰染色后,比较改良革兰染色法的染色效果。结果改良革兰染色法的染色效果好,该法具有易操作、染色效果稳定等优点。结论改良革兰染色法能够清晰地显示出组织切片中的真菌,并且染色效果稳定,在检测组织中的真菌时可发挥重要的辅助诊断价值,具有广泛的应用前景。     

六胺银染液配制方法的改进

六胺银 (Methenamie silver)染色广泛用于病理检测 ,是临床肾活检的重要的染色方法之一 ,该法可特殊地显示肾小球毛细血管基底膜 ,系膜基质、肾小囊壁层基膜以及肾小管基底膜的病理变化 ,亦是显示真菌、尿酸盐 [1 ] 等常用的一种方法。由于临床送检组织时多时少 ,预先配制的六胺银贮备液往往因放置过久而失效 ,导致染色失败。我们在 Gomor氏六胺银贮备液 [2 ] 的基础上将配制方法进行了改进 ,报道如下 :材料和方法1.试剂配制 :1.1　硝酸银分装于 EP管中 ,每只 2 5 mg,加盖 ,贴好标签 ,置 4℃冰箱内保存 ;六次甲基四胺 30 0 m g/只 EP管 ,…     

显示SARS病毒包涵体的技术探讨

SARS病毒包涵体是一种非常微小的微生物 ,在一般的情况下用光学显微镜是不易被发现的 ,但当它侵及细胞并形成小体时 ,则可用光学显微镜来检测 ,但较难寻找。为了寻找较有效的病毒包涵体染色方法来显示SARS病毒包涵体 ,我们对现有的病毒染色法如Macchiavello氏法的改良法、Mann氏法、Lendram氏法进行对照应用 ,经实验认为 ,Macchiavello氏的改良法效果最好 ,该法应用省时、易操作 ,可将病毒包涵体显示为鲜红色。至今为止 ,认为引起SARS的病毒是变性的冠状病毒 ,但由于该病毒非常微小 ,必须依靠电镜才能对其进行鉴定和鉴别。由于进入细…     

肺隐球菌病标本在六胺银染色中的最佳温育时间

目的探索肺隐球菌在六胺银染色中的最适当温育时间,以期获得最佳染色效果,提高病理标本肺隐球菌病的诊断率。方法收集厦门大学附属第一医院病理科2017年1月—2017年12月确诊的30例肺隐球菌感染病例,每例病例选取最具代表意义的一个蜡块连续切片3张,不同蜡块为区组因素,将每个蜡块切取的3张切片简单随机化分为3组,每组1张,通过区组随机化将90张切片随机分为A、B、C 3组,A组切片在六胺银染液步骤中采用水温箱60℃温育20～25min,B组切片在六胺银染液中温育30～35min,C组切片在六胺银染液中温育40～50min。以10分制对90张六胺银染色片进行评分,比较3组切片得分情况、显微镜下的染色效果与诊断准确率。结果 B组切片镜下显示:隐球菌染成空心黑色,细胞核呈蓝色,周围肺纤维组织呈淡棕色至棕色,隐球菌与周围肺纤维组织染色对比清晰,染色效果显著高于A组与C组。结论在六胺银染色中,采用六胺银染液在水温箱60℃温育30～35min,病理切片六胺银染色评分结果得分更高、六胺银染色效果更好、肺隐球菌病的诊断准确率更高。     

小肠上皮分泌细胞特性的组织学观察

目的寻找一种可以替代人体消化管的动物标本,并通过特殊染色方法,使得小肠上皮分泌细胞的形态特征能够明显地显示出来。方法随机采集成年猫小肠的新鲜标本,经Bouin液灌注固定24h后,石蜡包埋切片脱蜡入水。分别采用Gomori染色法、PAS反应、Gomori＋PAS反应、阿利新蓝（alcian blue,AB）染色法、AB＋PAS反应、HE染色法和苏木精-焰红染色法进行染色。结果在各种染色的切片标本上,能够观察到杯状细胞的形态、分布和染色特性以及肠内分泌细胞的特点,并发现在它们之间还存在一种绿色颗粒细胞和嗜酸性颗粒细胞。结论通过特殊染色可以肯定猫的小肠杯状细胞合成的是中性粘蛋白和酸性粘蛋白;绿色颗粒细胞为未成熟杯状细胞;嗜酸性颗粒细胞为Paneth细胞,其特点是单个分散分布。肠内分泌细胞与周围其他上皮细胞的染色对比明显而容易识别。     

应用超声波六胺银法的改良法显示组织胞浆菌

应用超声波六胺银法的改良法显示组织胞浆菌,效果好,图像清晰,细菌的菌壁被染成深黑色,菌体中央为空白区,背景为淡绿色。①该法需时少,六胺银液的浸染时间仅需２０分钟左右,而原法则需７０～９０分钟。②由于浸染时间短,减少了杂质的沉淀,增加了背景的清晰度。原法浸染时间长,常可产生背景的杂质沉淀,影响结果的评判。③该法利用超声波的高频振谐下所作的缩张运动,加速了溶液中分子的运动速度,从而促进了切片的氧化过程,使菌壁的醛基更加完全彻底的暴露出来,有利于还原溶液中的硝酸银成为金属银,将细菌给予更好地显示出来、从最后的结果证明：它优于原法或ＰＡＳ法。     

长鬣蜥脑垂体的组织学结构观察

用HE、尼氏染色和Holmes银染法对长鬣蜥脑垂体的显微结构进行了观察.发现其脑垂体分腺垂体、神经垂体两部分,为背腹型.HE染色观察到腺垂体分为嗜酸性细胞、嗜碱性细胞和嫌色细胞3种类型,另外还观察到室管膜细胞和神经垂体内的垂体裂;尼氏染色显示神经垂体的赫令氏体以及排列致密的腺垂体中间部细胞;Holmes银染法显示神经垂体的无髓神经纤维、垂体门脉系统的血管.首次发现垂体裂与中脑视叶的水管系统相连通.     

肾小球基底膜PASM染色方法的改进

在临床肾穿标本的病理诊断中,Gomori六胺银(PASM)染色是显示肾小球基底膜常用染色方法,在实际工作中,我们发现按常规的PASM染色法操作,染色时间长,肾小球毛细血管基底膜不易着色,且结构不清晰,背景欠干净,影响观察和诊断。为此,我们对几种方法进行比较,经反复摸索,发现利用超声波进行PASM染色具有操作简便、方法稳定、时间短、结构清晰和背景干净等优点,对提高制片质量和诊断很有帮助,现介绍如下。     

病理组织芽生菌感染的特殊染色方法分析

目的分析比较显示真菌的4种特殊染色方法 ,旨在寻找一种显示芽生菌并适合于临床病理诊断的染色方法。方法对病理石蜡组织芽生菌感染标本进行PAS染色、PASM染色、抗酸染色及爱尔新蓝染色(p H2.5),光学显微镜下观察染色效果。结果 HE染色切片上见多数圆形、厚壁、棕色的硬壳小体,PAS染色呈红色,PASM染色呈黑色,抗酸染色个别菌体呈红色,爱尔新蓝染色呈阴性。结论 PAS染色和PASM染色均可用于显示芽生菌,但改良后的PAS染色时间最短,结构清晰、对比明显,是检测芽生菌效果最佳的染色方法。     

基底膜染色方法比较及在胃粘膜异型增生与胃腺癌中的应用

目的探讨胃粘膜上皮基底膜有效、便捷、直观的实验方法,及基底膜、Ki67免疫组织化学、粘液多重染色在胃腺癌组织染色中的可行性。方法用过碘酸希夫(periodie acid Schiff,PAS)反应、六胺银法(methenamine silvermethod,PASM)、网状纤维染色、苦味酸-天狼猩红(Picrosirius red,SR)和IV型胶原免疫组织化学5种染色方法对多种胃粘膜异型增生及胃腺癌病理标本基底膜进行染色,并利用Friedman检验、Wilcoxon检验定量分析、比较基底膜阳性区域面积。此外,在同一张胃腺癌组织切片上进行显示基底膜、蛋白质、粘液的多重染色。结果 5种基底膜染色方法中,IV型胶原免疫反应阳性面积与SR染色阳性面积(胃粘膜中-重度异型增生标本除外)无显著性差异、PAS与PASM染色阳性面积无显著性差异、网状纤维染色阳性面积最大。多重染色实验结果显示胃腺癌Ki-67、阿尔新蓝(Alcian blue,AB)-PAS二重染色和Ki-67、AB、SR三重染色,分别显示对应结构,效果良好。结论观察基底膜,胃粘膜中-重度异型增生标本中IV型胶原免疫组织化学与SR染色要分别选用;胃粘膜轻度异型增生及胃腺癌标本中IV型与SR染色可相互替代;PAS与PASM染色法可相互替代,我们推荐较为便捷的SR和PAS染色;网状纤维染色观察比较直观,可选用。此外,SR、PAS也可分别应用于胃腺癌Ki67免疫组织化学与AB的多重染色中,效果良好。     

A Unique Protease Cleavage Site Predicted in the Spike Protein of the Novel Pneumonia Coronavirus (2019-nCoV) Potentially Related to Viral Transmissibility

正Dear Editor,In December 2019, a novel human coronavirus caused an epidemic of severe pneumonia(Coronavirus Disease 2019,COVID-19) in Wuhan, Hubei, China(Wu et al. 2020; Zhu et al. 2020). So far, this virus has spread to all areas of China and even to other countries. The epidemic has caused 67,102 confirmed infections with 1526 fatal cases     

Curcuminoids as inhibitors of thioredoxin reductase: A receptor based pharmacophore study with distance mapping of the active site

Curcumin is the yellow pigment of turmeric that interacts irreversibly forming an adduct with thioredoxin reductase (TrxR), an enzyme responsible for redox control of cell and defence against oxidative stress. Docking at both the active sites of TrxR was performed to compare the potency of three naturally occurring curcuminoids, namely curcumin, demethoxy curcumin and bis-demethoxy curcumin. Results show that active sites of TrxR occur at the junction of E and F chains. Volume and area of both cavities is predicted. It has been concluded by distance mapping of the most active conformations that Se atom of catalytic residue SeCYS498, is at a distance of 3.56 from C13 of demethoxy curcumin at the E chain active site, whereas C13 carbon atom forms adduct with Se atom of SeCys 498. We report that at least one methoxy group in curcuminoids is necessary for interation with catalytic residues of thioredoxin. Pharmacophore of both active sites of the TrxR receptor for curcumin and demethoxy curcumin molecules has been drawn and proposed for design and synthesis of most probable potent antiproliferative synthetic drugs.     

Comparative morphology of the carpel in the Liliaceae: Hewardieae, Petrosavieae, and Tricyrteae

The young pistils in the melanthioid tribes, Hewardieae, Petrosavieae and Tricyrteae, are uniformly tricarpellate and syncarpous. They lack raphide idioblasts. All are multiovulate, with bitegmic ovules. The Petrosavieae are marked by the presence of septal glands and incomplete syncarpy. Tepals and stamens adhere to the ovary in the Hewardieae and the Petrosavieae but not in the Tricyrteae. Two vascular bundles occur in the stamens of the Hewartlieae and Tricyrtis latifolia. Ventral bundles in the upper part of the ovary of the Hewardieae are continuous with compound septal bundles and placental bundles in the lower part. Putative ventral bundles occur in the alternate position in the Tricyrteae and putative placental bundles in the opposite. position in the Petrosavieae. The dichtomously branched stigma in each carpel of the Tricyrteae is supplied by a bifurcated dorsal bundle.     

Enterovirus 71 3C proteolytically processes the histone H3 N-terminal tail during infection

Highlights
1. The N-terminal tail of histone H3 is specifically cleaved during EV71 infection.
2. Viral protease 3C is identified as a protease responsible for proteolytically processing the N-terminal H3 tail.
3. Our finding reveals a new epigenetic regulatory mechanism for Enterovirus 71 in virus-host interactions.     

Detection of EBV and HHV6 in the Brain Tissue of Patients with Rasmussen’s Encephalitis

Rasmussen’s encephalitis (RE) is a rare pediatric neurological disorder, and the exact etiology is not clear. Viral infection may be involved in the pathogenesis of RE, but conflicting results have reported. In this study, we evaluated the expression of both Epstein-Barr virus (EBV) and human herpes virus (HHV) 6 antigens in brain sections from 30 patients with RE and 16 control individuals by immunohistochemistry. In the RE group, EBV and HHV6 antigens were detected in 56.7% (17/30) and 50% (15/30) of individuals, respectively. In contrast, no detectable EBV and HHV6 antigen expression was found in brain tissues of the control group. The co-expression of EBV and HHV6 was detected in 20.0% (6/30) of individuals. In particular, a 4-year-old boy had a typical clinical course, including a medical history of viral encephalitis, intractable epilepsy, and hemispheric atrophy. The co-expression of EBV and HHV6 was detected in neurons and astrocytes in the brain tissue, accompanied by a high frequency of CD8⁺ T cells. Our results suggest that EBV and HHV6 infection and the activation of CD8⁺ T cells are involved in the pathogenesis of RE.     

Perinatal Vertical Transmission of Chikungunya Virus in Ruili,a Town on the Border between China and Myanmar

正Dear Editor,Chikungunya virus (CHIKV), an arbovirus in the family of Togaviridae, genus Alphavirus, is transmitted by the A.aegyptii or A. albopictus mosquito, and causes disease in humans characterized by fever, rash, and arthralgia (Silva and Dermody 2017; Suhrbier 2019). It was first reported in 1953 in Tanzania, and caused only a few outbreaks and sporadic cases in Africa and Asia in last century. However, in the epidemic in 2004, CHIKV acquired mutations that conferred enhanced transmission by the A. albopictus mosquito(Schuffenecker et al. 2006). Since then, it has successively caused outbreaks in Africa, the Indian Ocean, South East Asia, the South America, and Europe (Zeller et al. 2016).     

Development of a Novel Reverse Transcription Loop-Mediated Isothermal Amplification Method for Rapid Detection of SARS-CoV-2

In conclusion, the novel visual RT-LAMP assay is a simple, rapid, and sensitive approach for detection of SARS-CoV-2, and it is ready for application in primary care and community hospitals or health care centers, and even patients' own houses in response to the current SARS-CoV-2 epidemic because the assay does not require sophisticated equipment and skilled personnel. Furthermore, it is also ready to be used in fields for screening samples from wild animals and environments to facilitate the identification of potential intermediate hosts that mediate the cross-species transmission of SARS-CoV-2 from bats to humans.