Golgi-like demonstration of “dark” neurons with an argyrophil III method for experimental neuropathology

Summary A silver method is proposed for the selective, well-contrasted and reproducible demonstration of dark neurons in frozen, vibratome and paraffin sections cut at a thickness of 5 to 200 m from aldehyde-fixed brains. The Golgi-like staining of the dendrites enables asorting of dark neurons according to characteristic neuron classifications. The staining procedure includes an esterification with 1-propanol, a treatment with diluted acetic acid and development. The esterification strongly increases the argyrophilia of both dark neurons and mitochondria. Unwanted co-staining of mitochondria is suppressed by the acetic acid treatment, while a special developer is used to render the staining controllable. The applicability of the method to experimental neuropathology is demonstrated by Golgi-like staining of dark neurons in rat brains exposed, before transcardial perfusion-fixation and delayed autopsy, to various pathological conditions including ischemia, hypoglycemia, trauma, status epilepticus, deafferentation and poisoning with kainic acid, colchicine and sodium azide, respectively.     

203例幽门螺杆菌相关性胃病的临床病理诊断分析

目的:探讨基层医院胃、十二指肠疾病幽门螺杆菌(HP)感染情况,检测方法及HP感染性胃病的临床病理形态特点。方法:采用北京同仁医院消化内科刘宾等提供的改良胃镜下HP感染的可视化诊断和Giemsa染色的HP形态学病理诊断方法,对203例上胃肠道疾病患者进行检测,并对其胃粘膜进行组织形态观察。结果:改良胃镜下HP的可视化诊断阳性率为96.55％,Giemsa染色病理诊断阳性率为95.07％,改良胃镜下HP感染的可视化诊断与病理诊断符合率为98.47％。HP感染胃粘膜有特征性组织形态改变。结论:改良胃镜下HP感染的可视化诊断及Giemsa染色HP形态学病理诊断相结合运用于胃、十二脂肠疾病的HP检测,适合普通基层医院采用。HP感染胃粘膜有特征性组织形态改变。     

银染法与放射自显影法检测醛糖还原酶基因多态性的比较

目的 :探讨银染法检测醛糖还原酶 (aldosereductase,AR)基因 5′端微卫星多态性的可靠性。方法 :用银染法和放射自显影法分别检测相同样本DNA的AR基因微卫星多态性 ,比较二者结果。结果 :两种方法均能清楚判读基因型且结果一致。结论 :银染法可以代替放射自显影方法检测AR基因的微卫星多态性     

食管粘膜活检取样误差的发生机制及对策

目的：探讨食管粘膜活检取样误差的发生机制及对策。方法：对来自河南食管癌高发区２８例患者行食管内镜检查和中、下段粘膜活检组织病理检查,１０天短期随访后对同一人群作重复检查;并对８８位无症状受检者进行碘染与活检相结合的方法。结果：第二次粘膜活检组织中,食管中段２４％的患者病变减轻,２８％的患者病变加重,４８％的患者病变维持不变。食管下段粘膜活检中,４８％的患者病变减轻,１６％的患者病变加重,３２％的患者病变维持不变;碘染受检者的食管粘膜中染色正常和染色异常的粘膜组织在癌前病变各阶段的构成比无明显差异。组织形态学测量显示：各级癌前组织与正常上皮组织的厚度无明显差异。结论：粘膜活检取样误差可严重影响组织病理学检查随访结果。食管粘膜癌前病变,特别是较轻度病变的厚度与同期正常上皮无明显改变,造成内镜下识别困难,是造成取样误差的重要因素。碘染可明显提高内镜下病变与同期正常组织的识别,但对小范围病变的活检准确度仍需改进。肿瘤抑制基因ｐ５３蛋白聚集的重复性检出率较高,可能是食管癌变过程中的有效指标。     

单宁酸——氯化铁法媒染心脏微血管的研究

应用单宁酸－氯化铁法媒染大鼠心脏微血管,光镜下观察其形态学特点。结果：左、右心室及空间隔、心房的微血管得到了良好的显示。左、右冠状动脉细小分支在心外膜层、心肌层及心内膜层形成了三层血管网,内、外两层微血管互相吻合成网;心肌层微血管则与心肌纤维平行,很少吻合。各级微血管的形态自然真实,有明显立体感。因此,单宁酸－氯化铁法是显示心脏微血管的理想方法。     

锥兰联合茜素红活细胞染色法观察角膜中央碱烧伤内皮细胞愈合过程

目的:研究角膜碱烧伤内皮细胞的愈合过程。方法:锥兰联合茜素红内皮细胞染色法对角膜中央碱烧伤内皮细胞形态学变化进行观察。结果:伤后20分钟内皮细胞已破坏。72小时,损伤区的周边内皮细胞变形向创面内迁移。第7天后缺损区大部分范围均可见梭形细胞覆盖。结论:兔碱烧伤损伤区内皮细胞的修复由邻近内皮细胞变形,增殖和移行长入创面内完成,修复的内皮细胞具有纤维细胞特征。眼科学报1999;15:218-220。     

A prospective study to evaluate the efficacy of modified swim-up preparation for male sex selection

A previously published study reporting the use of a modifiedswim-up technique for sperm preparation prior to inseminationwhich resulted in a high percentage of male births has beencriticized for its lack of controls. The present prospectivestudy was initiated to investigate further the efficacy of modifiedswim-up preparation for male sex-selection when applied in aproperly defined control group. Our results showed that theproportion of males born in singleton pregnancies was 50% inthe group inseminated following sperm preparation with Percolland in the control group with no sperm preparation comparedwith 88.5% in the group treated with the modified method ofswim-up sperm preparation prior to insemination. This high rateof males in the group treated with modified swim-up was alsoobserved in singleton pregnancies of women taking ovulationinducing drugs (primarily clomiphene citrate). This contrastswith previous publications in which a higher rate of femaleswas found in clomiphene citrate patients using the albumin separationtechnique. How the mechanism of the swim-up procedure may resultin a high male birth rate remains unclear. A high percentageof y enriched semen was found using fluorochrome quinacrinemustard staining but this procedure may falsely stain autosomalchromosomes. If analysis using sensitive DNA probes fails toconfirm the y enrichment of the spermatozoa, one must hypothesizethat the modified swim-up procedure damages the x-spermatozoa.     

α-Smooth-muscle actin expression in normal and fibrotic human livers

To determine the significance of the expression of -smooth-muscle actin in the fibrotic human liver, normal and diseased livers were stained with anti--smooth-muscle-actin antibody by an immunoperoxidase method. Vitamin A-containing lipocytes were also identified by the modified Kupffer's gold chloride method. In the normal human liver, lipocytes as well as vascular smooth muscle cells expressed -smooth-muscle actin. In alcoholic liver disease, there was an increase in the cells positive for -smooth-muscle actin adjacent to the fibrotic areas, but the response of lipocytes to the gold chloride reaction diminished. In chronic hepatitis, the cells positive for -smooth-muscle actin increased around the enlarged portal areas, and the response to the gold chloride reaction did not change appreciably. An increase in the cells positive for -smooth-muscle actin was associated with the progression of hepatic fibrosis in the liver of patients with alcoholic liver disease and chronic hepatitis.     

Primary culture of vital marginal cells from cochlear explants of the stria vascularis

Summary Explants of the stria vascularis and spiral ligament were dissected from guinea pig cochleae and were successfully cultivated for several weeks. After 2 days, fibroblast-like cells of the spiral ligament covered the bottom of the cell culture dish around the explant. Marginal cells of the stria vascularis proliferated and grew on the luminal surface towards the border of the explant at a rate of 15 m/day. At day 6 in culture the proliferating marginal cells reached the border of the explant and then advanced to the bottom of the cell-culture dish. There the marginal cells replaced fibroblast-like cells and built an epithelial hexagonal-shaped monolayer. Light microscopic and transmission electron microscopic investigations revealed that the cultured cells were viable and that typical morphological characteristics of marginal cells were preserved. Cultivation of these cells provides a unique model for studies of physiological properties of marginal cells of the stria vascularis.     

Bulbectomy Enhances Neurogenesis and Cell Turnover of Primary Olfactory Neurons But Does Not Abolish Carnosine Expression

Primary olfactory neurons located in the olfactory neuroepithelium project to the ipsilateral olfactory bulb and undergo a continuous process of neurogenesis and differentiation. We describe, in the adult rat, the kinetics of proliferation, differentiation and survival of primary olfactory neurons either in the presence or absence of their target, the olfactory bulb. The experimental design included unilateral bulbectomy, coupled with a single bromodeoxyuridine pulse 35 days after surgery. The rate of proliferation and survival of olfactory neurons was then examined by immunohistochemistry for bromodeoxyuridine, and the differentiation status by in situ hybridization for calmodulin messenger RNA in immature and mature olfactory neurons and immunohistochemistry for the dipeptide carnosine in mature olfactory neurons. We show that primary olfactory neurons can synthesize carnosine in the absence of the olfactory bulb. However, the number of carnosine-immunopositive neurons in the absence of their target is dramatically reduced to less than one-fourth, whereas the number of olfactory neurons expressing calmodulin messenger RNA is only slightly reduced. The numeric reduction of camosine-positive neurons in the target-deprived neuroepithelium is correlated with a dramatic reduction in the survival rate of olfactory neurons, since newly generated olfactory neurons are completely lost 35 days after the bromodeoxyuridine pulse. In contrast, in the normal olfactory neuroepithelium almost one-third of newly generated olfactory neurons survive 35 days after the bromodeoxyuridine pulse. On the whole, these data indicate that most of the primary olfactory neurons have a short lifespan but that once they have connected with the olfactory bulb they may persist longer, and suggest that throughout adulthood olfactory neurons are overproduced, differentiate independently from their target, and then undergo a process of target-induced neuronal selection.