同时显示肾微血管和细胞构筑的实验研究

目的：在媒染微血管的同时显示肾内细胞构筑，以了解肾微血管与肾小管之间的相互关系。方法：用单宁酸一氯化铁法(TA—be)媒染肾微血管，再以苏木精、H—E法复染来观察肾小管细胞结构。结果：经复染的2组切片各级血管和细胞均染色良好，血管壁平滑肌纤维、内皮细胞界限清晰，细胞核蓝染，H—E染色的细胞质呈深浅不同的红色。因切片较厚，保证了微血管的立体感，细胞的染色效果却不及薄切片的鲜艳。结论：TA—Fe法配合苏木精或HE复染，可同时显示肾微血管和细胞构筑。     

单宁酸-氯化铁法媒染肺内微血管及细胞的组织学观察

目的　光镜下观察大鼠肺微血管及肺组织细胞。方法　应用单宁酸 氯化铁法 (TA Fe法 )灌流固定大鼠 ,取肺做冰冻切片 ,氯化铁显色 ,光镜下观察。结果　肺内小血管、微血管蜿蜒走行 ,被切成各种断面 ,管壁平滑肌细胞、内皮细胞显示清晰 ,有一定立体感 ;接近被膜的微动脉形成较粗的毛细血管 ,肺泡间毛细血管不易分辨 ,肺间质内有微血管和毛细血管分布 ,微血管周围的肺组织亦被媒染。结论　TA Fe法即可显示肺内微血管 ,又能保存组织细胞结构 ,肺内小动脉和微动脉管壁的平滑肌参与微循环的主动调节机制。     

改进前后TA-Fe法媒染肾微血管和肾小管效果的对比

应用TA-Fe法媒染肾微血管[1,2]，为肾微血管构筑和临床有关疾病的诊断和研究提供了一种简便易行的新方法，但是因其需湿片摄影，时间要求严格。而自然干燥后切片易收缩卷曲、脱落，无法保存原始材料进行回顾性研究。为此，我们对TA-Fe法进行了改进，结果令人满意。它既清晰地媒染微血管，又良好显示了肾微血管与肾小管的关系，切片可长时间保存。     

改良单宁酸-氯化铁法媒染子宫血管的实验研究

目的：光镜下观察大鼠子宫各部位血管构筑。方法：应用单宁酸一氯化铁法(TA—Fe法)灌流固定大鼠，取子宫不同位置做冰冻切片，氯化铁显色，常规脱水、透明、封片，显微摄影。结果：子宫壁出现类似电镜负染色效果，组织结构灰黑色，血管双线条状，分支明显，过管壁切面可见内皮细胞或平滑肌；子宫动脉进入肌层分支并形成一、二级血管网和多支环状动脉，环行动脉或二级血管网发出的微动脉和毛细血管进入粘膜，形成密集的三级毛细血管网。结论：经灌流的子宫标本较长时间保存在媒染固定液内，导致血管外结构也被媒染；子宫壁有三级血管网；每个子宫有多支环状动脉；各段子宫血管构筑不尽相同。     

TA-Fe法媒染骨骼肌微血管及肌纤维的光镜研究

目的　光镜下观察骨骼肌微血管构筑及其与肌纤维的关系。方法　应用单宁酸媒染固定液灌流大鼠 ,取髂腰肌 ,冰冻切片 ,氯化铁显色 ,光镜下观察。结果　髂腰肌内微动脉以一定角度进入肌束内 ,形成毛细血管前微动脉或毛细血管营养纤维 ,毛细血管可同向走行 ,亦可呈反向或爪形分支 ,常见两条或者几条毛细血管互相吻合成网 ,多数毛细血管长距离直行于肌纤维之间 ,偶见波浪状走行者 ,毛细血管呈双线条状 ,可见细胞核 ,肌纤维保存完好 ,横纹清晰。结论　TA Fe法可清晰的显示髂腰肌微血管和肌纤维 ,血管内皮细胞和肌细胞的核明显     

应用单宁酸的负染色效果显示大鼠生殖器官微血管

近百年来,许多学者为研究器官内血管立体构筑倾注了大量心血,创造了多种显示微血管构筑的方法,诸如墨汁灌注法,橡胶、维尼龙血管、淋巴管立体铸型法,微血管铸型扫描电镜法,碱性磷酸酶法（钙铅法）以及单宁酸-氯化铁媒染法（TA-Fe法）,为科学研究及临床应用做出了贡献。继TA-Fe法媒染微血管后,我们又发现单宁酸过度媒染某些组织结构,有类似电镜标本负染色技术效果,清晰地显示了大鼠子宫、卵巢、输卵管等器官的血管构筑特点。     

犬肾血管构筑及血管和肾小管相互关系——扫描电镜观察

使用同时显示肾血管和肾小管扫描电镜法及肾血管铸型扫描电镜法,观察研究了犬肾血管构筑及肾血管和肾小管相互关系。血管球内存在亚小叶微循环单位;血管球内存在直捷通路;迷路部血管和肾小管无明显的逆流配置;球后血管在肾皮质内有广泛吻合;肾髓质次级血管束内一些降直血管具有门静脉特点;直血管在乳头部肾盂上皮下形成致密血管丛,它是升、降直血管间的主要连接通道;内髓升、降直血管间还有毛细血管及少量血管襻连接。本文还讨论了上述结构特点的功能意义。     

明胶墨汁与荧光微球显示大鼠视网膜微血管灌流效果的对比研究

目的:比较明胶墨汁灌注与荧光微球灌注对大鼠视网膜微血管灌流情况的显示效果。方法:12只成年健康SD大鼠随机分成荧光微球组(6只)和明胶墨汁组(6只)。明胶墨汁组分两步经左心室灌注明胶墨汁40ml。荧光微球组经股静脉注射荧光微球0.5ml。视网膜行铺片和切片,观察荧光微球和改良墨汁灌注显示的微血管灌流情况;两组视网膜切片还进行NeuN、Parvalbumin和GFAP的免疫组化染色。结果:荧光微球零散分布于视网膜铺片周边部和中央部视的网膜血管中;不能显示铺片的血管轮廓和切片中的血管截面。荧光微球与NeuN、Parvalbumin或GFAP免疫组化双标不能在同一组织中准确显示视网膜血管与神经细胞的紧密空间关系。改良墨汁灌注能清晰显示大鼠视网膜铺片中周边部和中央部的血管网,以及切片中的血管截面,而且此方法与NeuN、Parvalbumin或GFAP免疫组化双标能在同一组织中准确显示视网膜血管与神经细胞的紧密空间关系。结论:改良明胶墨汁灌注在显示大鼠视网膜微血管灌流情况方面优于荧光微球,适宜于在视网膜疾病研究中广泛使用。     

TA-Fe法显示微血管及大脑不同区域微血管的定量研究

目的　定量分析研究大脑不同区域微血管的直径、密度及分布。方法　应用TA Fe法媒染大鼠大脑微血管 ,光镜下观察其形态结构 ;用目镜测微尺按体视学方法测定大脑相关区域微血管直径、密度 (MVD) ;用CMIAS图像分析系统测定微血管面积比 (MVA )。结果　血管直径 :顶、枕、颞叶皮质差别不明显 (P >0 .0 5 ) ,皮质与髓质、内囊均有显著性差异 (P<0 .0 5 )。MVD、MVA值 :顶区皮质 >枕区皮质 >颞区皮质 >髓质 >内囊 ,皮质间MVD无明显差别 ,皮质各区与髓质、内囊均有显著性差异。结论　TA Fe法是一种研究微血管构筑的理想方法 ,大脑各区皮质内MVD、MVA基本相同 ,皮质明显高于髓质和内囊。     

多种金属离子与单宁酸反应媒染微血管的对比研究

目的　镜下观察单宁酸与金属盐溶液中Ca2 + 、Au+ 、Ag+ 、Pb2 + 、Cu2 + 、Al3 + 、U6+ 、K+ 联用显示大脑微血管的效果。方法　用单宁酸媒染固定液灌流大鼠 ,取脑切片 ,入氯化钙、氯化金、硝酸银、硝酸铅、硫酸铜、硫酸铝钾、醋酸双氧铀、高锰酸钾和重铬酸钾等溶液中呈色。结果　单宁酸与Ca2 + 、Cu2 + 、Ag+ 、Al3 + 结合显示血管清晰 ,与Au+ 、U6+ 、Pb2 + 、K+ 联用媒染血管效果欠佳。结论　含Ca2 + 、Cu2 + 、Ag+ 、Al3 + 的金属盐类可替代氯化铁媒染微血管 ,氯化金、醋酸双氧铀、硝酸铅、重铬酸钾和高锰酸钾不宜与单宁酸联用来显示血管     

Histochemical Analysis of Renal Dysplasia with Ureteral Atresia

Unilateral small kidney with ureteral obstruction was discovered in a 74-year-old female cadaver during an anatomical dissection course. In order to elucidate the histogenesis of renal dysplasia, we carried out histochemical and immunohistochemical analyses. On macroscopic view, the kidney was approximately 3 cm in length, 2 cm in width and weighed only 9 g. Although the ureter ran from the renal hilus to the bladder, its width was under 2 mm. The renal parenchyma was extremely thin and there was a large congested vein in the renal sinus. On microscopic examination of the kidney, we observed that numerous developing renal tubules had cytokeratin-positive epithelia, most of which were surrounded by concentric fibrosis. However, we could not detect any structures resembling the collecting duct, renal tubules, renal pelvis, or glomeruli. The concentric mesencymal fibrous tissue surrounding the immature renal tubules contained the smooth muscles that were positive for h-caldesmon. Serial sections of the ureter revealed several small and discontinuous lacunae lined by cuboidal and transitional epithelium, which did not constitute a patent lumen through the bladder. This case is a rare case of renal dysplasia with defect in recanalization of the ureteral bud during the early developmental stage.     

Potential pitfalls in the frozen section evaluation of parenchymal margins in nephron-sparing surgery

With advances in radiographic imaging, there has been an increase in the incidental detection of small renal cell carcinomas, with a resultant increase in partial nephrectomies for these tumors. Partial nephrectomy often necessitates assessment of renal parenchymal margins by frozen section. To determine the most common problematic "lesions" encountered on renal parenchymal margins, we evaluated all diagnostically challenging frozen sections that had been referred to a genitourinary pathologist. Frozen sections with detached atypical cells and crushed tubules were the most common lesions that presented diagnostic uncertainty. We found that normal constituents of renal parenchyma, namely tubules and glomeruli, can be mistaken for neoplasia. Neoplastic tubules of low-grade renal cell carcinomas may be misinterpreted as thickly cut, crushed benign tubules, and the significance of tubulopapillary "adenomas" in frozen sections is unclear. The present report highlights diagnostic difficulties that pathologists may encounter on frozen sections of renal parenchymal margins.     

根据肾小管轮廓计数和直径估计肾小管长度的两种体视学方法对比

目的 对比估计肾小管长度的两种体视学方法：轮廓计数法和直径法。方法 从2月龄正常雄性SD大鼠获取9个肾脏，采用定向法制作等向随机切片（甲基丙烯酸树脂包埋），用过碘酸-雪夫（PAS）试剂和苏木素染色后进行体视学测量：计数切片上各段肾小管（包括集合管，不包括髓袢细段）轮廓（切面）中心管腔的数量，由此估计肾小管的长度；抽选近似圆形、椭圆形、有明显管腔的肾小管轮廓，测其直径（圆形轮廓）或短径（椭圆形轮廓），据其平方的平均计算肾小管（假设为圆柱结构）横断面积，然后结合肾小管的体积估计其长度。结果 据轮廓直径估计的各个肾脏内肾小管的总长与据轮廓计数估计的结果之比，平均为1.43（变异系数7.2%），即轮廓直径法高估了大于40%的肾小管长度。结论 用肾小管的短径来估计其长度的方法不可靠。     

Renal cell tumors induced in CBA male mice by 1,2-dimethylhydrazine.

Epithelial kidney tumors induced in CBA male mice by 1,2-dimethylhydrazine (DMH) were studied histochemically and immunohistochemically. A total of 48 tumors studied histologically were diagnosed as clear-cell, acidophilic, or mixed adenomas located in the renal cortex. Gamma-glutamyl transpeptidase (GGT) was strongly positive in normal proximal convoluted tubules, slightly positive in the cells of Bowman's capsule, and negative in 47 of 48 tumors examined. Antibodies against the new antigen obtained from the mouse liver oval cells, antigen A6, were also used. This antigen is negative in normal kidney proximal tubules but always positive in distal tubules and collecting ducts. It was also positive in all 47 GGT-negative tumors studied here. One tumor was GGT-positive and antigen A6-negative. Based on our data, it was concluded that the majority of renal cell adenomas induced by DMH in mice probably originate from the distal tubules or collecting ducts and not from the proximal tubules.     

Demonstration of Actomyosin in Mesangial Cells of the Renal Glomerulus

Antisera to human uterine actomyosin were prepared in rabbits and conjugated with fluorescein (F-AUAM). When F-AUAM was applied to frozen sections of normal human kidney which were then examined by ultraviolet light microscopy, it was observed that vascular smooth muscle, endothelium of arteries, veins, and peritubular capillaries and glomerular mesangial cells were immunofluorescent. Neither glomerular endothelium nor epithelial cells of Bowman's capsule or renal tubules were stained by F-AUAM. The specificity of antisera for actomyosin was confirmed by absorption and blocking studies, examination of a wide variety of tissues and immunodiffusion in agarose gel. It may be inferred from these data that mesangial cells are contractile. Contraction of the mesangium may play a significant role in regulating glomerular blood flow and in the reaction of the glomerulus to injury.     

Renal tubular dysgenesis: delayed onset of oligohydramnios

Two premature sibs had Potter sequence and died of respiratory failure within the first day. Ultrasonography at 26 weeks during the earlier of the two pregnancies showed complete absence of amniotic fluid, and the urinary bladder was not visualized. Ultrasound examinations during the second pregnancy showed adequate amniotic fluid at 16 and 20 weeks, with a subsequent reduction in fluid volume. Two older sibs had also died of respiratory failure shortly after birth. Postmortem histopathologic studies showed all four sibs to have severely deficient renal tubular development. However, the presence of numerous glomeruli indicated prolific nephrogenesis. Most of the tubules in sections of cortex had the lectin-binding and immunohistochemical characteristics of collecting ducts; proximal tubules were not identified by lectin-binding. Electron-microscopic examination showed a general absence of differentiated characteristics in cortical tubular epithelium, except that rare tubules contained rudimentary proximal tubular brush borders. Three of the sibs were boys, one a girl. The three children that were studied had normal chromosomes. Two unaffected sibs are alive and well. Neither parent has any clinical evidence of renal disease. These studies support the interpretation that renal tubular dysgenesis is autosomal recessive with pleiotropy. However, the relatively late appearance of oligohydramnios makes early diagnosis difficult, even when the condition is suspected.     

四氯化碳诱导大鼠肾细胞凋亡的研究

目的观察四氯化碳（CCl4）诱导大鼠肾形态结构和肾细胞凋亡的变化。方法将健康雄性Wistar大鼠随机分为对照组、2周CCl4组和6周CCl4组,每组5只。对照组背部皮下注射橄榄油（0．12ml／100g鼠重）,CCl4组以0．3ml／100g鼠重背部皮下注射60％CCl4橄榄油。2周CCl4组和6周CCl4组大鼠分别于实验的第3周和7周的第1d处死。肾石蜡切片苏木精．伊红染色,光镜观察肾组织形态学,免疫组织化学显示Caspase-3、Bax、TUNEL观察肾细胞凋亡。结果对照组肾小球和肾小管结构均正常。2周和6周CCl4组的肾小球结构正常,2周CCl4组病变较轻、6周CCl4组肾皮质近端小管管腔狭小,上皮细胞浊肿,空泡变性,微绒毛脱失。对照组和2周CCl4组Caspase-3、Bax免疫组织化学显色未见阳性细胞表达;而6周CCl4组则在近髓质处的皮质内,近端小管上皮细胞的胞质呈现棕黄色阳性反应。TUNEL显色2周CCl4组可见肾皮质内近端小管上皮细胞核和少量的肾小球细胞核呈阳性表达,而6周CCl4组在大量的远曲小管及少量近端小管的上皮细胞核呈阳性表达。结论CCl4诱导性肾损伤大鼠,2周时肾组织的近端小管上皮及少量肾小球毛细血管内皮细胞发生凋亡,而损伤6周时,肾皮质近端小管受损,上皮细胞变性;远曲小管上皮细胞发生凋亡。     

Overview Article: Basal Lamina of Epidermis,Muscle Fibers,Muscle Capillaries,and Renal Tubules: Changes with Aging and in Diabetes Mellitus

Using autopsy material from 14 diabetics and 10 controls (8–84 years old), the authors measured the thickness of basal lamina (BL) in skeletal muscle capillaries, renal tubules, skeletal muscle fibers, and epidermis to determine whether BL accumulation is a generalized phenomenon or limited only to certain anatomic structures. The four structures were selected because earlier experiments in animals have shown that in two (muscle capillaries and renal tubules) BL accumulates as a by-product of cell renewal while in the other two (muscle fibers and epidermis) it does not. In human tissues we found that 1) BL accumulates in muscle capillaries and renal tubules but not in muscle fibers and epidermis, 2) in muscle capillaries and in renal tubules it accumulates in controls and in diabetics as a function of aging, 3) more BL in both anatomic structures accumulates in diabetics than in controls, and 4) the extent of BL accumulation in muscle capillaries and renal tubules does not correlate with duration of diabetes mellitus. In addition to the fact that BL does not accumulate in all anatomic structures in which BL is normally present, the observations indicate that 1) diabetes alone is not responsible for BL accumule tion, 2) diabetes exaggerates age-dependent accumulation of BL, and 3) accumulation of BL in man is probably a by-product of cell renewal.