Comparison of methods for staining microvessels in bone

Abstract

Detection of microvessels is critical for studying bone tissue. We developed an intravascular ink-based method coupled with Van Gieson (VG) staining and compared it with other commonly used methods for capillary visualization. The ink perfusate was formulated as 10% ink, 10% formaldehyde and 20% mannitol. The ink solution was perfused into a healthy goat and the tibia was subjected to decalcification, dehydration, paraffin embedding, de-waxing and staining to observe microvessels. Angiogenesis was assessed by vascular area image analysis and the hematoxylin and eosin (HE), Masson, and VG staining techniques were compared to determine the reliability of these methods for counting microvessels. We found that HE, Masson, and VG staining produced poor contrast between the microvessels and surrounding tissues. By contrast, ink coupled with VG staining permitted clear discrimination between the microvessels and surrounding tissues. Our results indicate that ink-VG staining could be more useful than other methods for detecting tissue microvessels.     

Comparison of verdeluz orange G and modified Gallego stains

Tumors of the oral cavity include combinations of hard and soft tissues that may be difficult to identify using routine hematoxylin and eosin (H & E) staining. Although combination stains can demonstrate hard and soft tissues, trichrome stains, such as VanGieson and Masson, cannot differentiate dental hard tissues, such as dentin, cementum and osteoid. Modified Gallegos (MGS) and verdeluz orange G-acid fuchsin (VOF) stains can differentiate components of teeth. We used 10 tissue sections of decalcified bone and 10 pathologic tissue sections that contained different calcified tissues including peripheral ossifying fibroma, odontoma, central ossifying fibroma and cemento-ossifying fibroma. Sections were stained with H & E, VOF or MGS. H and E stained both hard tissues pink. VOF stained bone purple-red, cementum red and collagen blue. MGS stained bone green-blue, cementum red and collagen blue. VOF staining intensity and differentiation was better than MGS staining. VOF staining demonstrated hard tissue components distinctly and exhibited good contrast with the surrounding connective tissue. VOF also is a simple, single step, rapid staining procedure.     

Evaluation of histological techniques for the detection of fungal infections caused byLeptolegnia chapmanii (Oomycetes: Saprolegniales) inAedes aegypti (Diptera: Culicidae) larvae

We evaluated which of the fixatives and stains most frequently used for observation of insect tissues were the most appropriate for histopathological visualization of entomopathogenic fungal infections with Leptolegnia chapmanii in larvae of Aedes aegypti. The best contrast between the host tissues and the fungal structures was obtained when using a combination of Camoy fixative with Grocott staining contrasted with light green. Masson trichromic stain combined with 10% formaldehyde-phosphate buffer also provided satisfactory results--a good contrast and clearly distinguishable host tissues and fungal structures.     

Differentiation of bone and soft tissues in formalin-fixed, paraffin-embedded tissue by using methylene blue/acid fuchsin stain

OBJECTIVE: To find a staining method for formalin-fixed, paraffin-embedded tissue that would distinguish bone from surrounding soft tissues, including muscle, periosteal tissue and bone marrow. STUDY DESIGN: A variety of stains were tested and compared with hematoxylin-eosin. The potential value of any given stain was evaluated based on its ability to stain bone and soft tissues different colors or shades that could be readily identified in photomicrographs. Stains were evaluated using both endochondral (tibia) and intramembranous bone (calvaria) samples. RESULTS: In contrast to standard hematoxylin-eosin stain, which stains both bone and soft tissues pink, the methylene blue/acid fuchsin stain demonstrates remarkable contrast between bone and other tissues. Methylene blue/acid fuchsin stained bone bright pink and the surrounding soft tissues blue-purple. CONCLUSION: In addition to the superior staining properties of methylene blue/acid fuchsin, other benefits of this stain include its stability, ease of use and low cost. This stain has many potential applications in the study of erosive bone disease in humans and also in animal models for research.     

Comparison of tetrachromic VOF stain to other histochemical staining techniques for characterizing stromal soft and hard tissue components

The components of hard tissues including dentin, enamel, cementum, bone and other calcified deposits, and mature and immature collagen pose problems for identification in routine hematoxylin and eosin (H & E) stained sections. Use of combinations of stains can demonstrate the components of hard tissues and soft tissues distinctly. We assessed the efficacy of the Verde Luz-orange G-acid fuchsin (VOF) stain for differentiating hard and soft connective tissues and compared results with other histochemical staining techniques. Eighty tissue sections comprising developing tooth (30), ossifying fibroma (30) and miscellaneous pathologies (20) expected to contain varying types of calcified tissues were stained with H & E, VOF, and Masson's trichrome (MT). In developing tooth, VOF demonstrated better differentiation of hard tissues, while it was comparable to MT for ossifying fibroma and miscellaneous pathologies. The intensity of staining was greater with VOF than with the other stains studied. VOF stains hard tissue components distinctly and gives good contrast with the surrounding connective tissue. VOF is comparable to MT, but has added advantages including single step staining, rapid and easy procedures, and it distinguishes the maturity of the tissues.     

A cardiac tissue-specific binding agent of troponin I

A fluorescently labeled, biphenylalanine-rich peptide was identified as a should be cardiac troponin I-specific binding agent with preferential staining affinity to myocardium tissues and extremely low staining to other stromal components. Fluorescence images demonstrate that this new peptide is an excellent contrast agent useful for examining troponin I structural distribution and expression density within heart tissue sections. It provides a clear contrast between myocardial cells and the surrounding collagen matrix.     

通络益气方对博来霉素致大鼠肺纤维化的治疗作用研究

目的:研究通络益气方对博来霉素致大鼠模型肺组织病理及肺组织羟脯氨酸(HYP)的影响。方法:将36只SPF级健康6周龄Wistar雄性大鼠置于SPF级条件下,适应性试养3天后,分为3组:空白组、模型组、治疗组,每组12只。除空白组外,其余两组,在清醒状态下,放入与小动物雾化给药仪相连的30 cm×30 cm×20 cm自制玻璃箱中,雾化吸入浓度为5 g/L(50%)的博莱霉素,雾化20分钟,休息5分钟,一天连续激发2小时,连续刺激4周。空白组在同样条件下雾化吸入0.9%生理盐水,操作方法与其余两组相同。治疗组大鼠造模后第2天起给予通络益气方,通络益气方溶液按体质量给予大鼠药液1 m L/100 g灌胃,每次于雾化前1h灌胃;模型组、正常组均给予生理盐水(1 m L/100 g)灌胃。各组大鼠连续刺激4周后,用1%戊巴比妥钠(50 mg/kg)麻醉后,待其肌肉松驰、呼吸平稳后,仰面固定于专用板上,用动物专用CT进行肺扫描。各组大鼠在末次诱导后,24 h内腹腔注射1%戊巴比妥钠麻醉后,取肺组织进行HE和Masson染色,观察各组大鼠肺组织病理HE染色、Masson染色及肺组织中HYP的含量。结果:与空白组(0.76±0.06)相比,治疗组(1.11±0.13)、模型组(1.47±0.22)HYP均升高(P<0.05);与模型组(1.47±0.22)相比,治疗组(1.11±0.13)HYP明显降低(P<0.05)。结论:通络益气方能降低对肺纤维化大鼠肺组织中胶原的沉积,起到治疗肺纤维化的作用,与通络益气方抑制HYP胶原蛋白的表达有关。     

多种特殊染色法在骨关节炎组织形态学研究中的应用比较

目的探讨多种特殊染色法在骨关节组织中的染色规律及其在骨关节炎形态学研究中的应用价值。方法 6月龄健康新西兰大白兔20只,随机分为正常组和造模组各10只,根据改良Hulth法造模,6周后膝关节取材。对标本固定、脱钙后进行石蜡包埋和切片。分别采用HE、番红-固绿、AB-PAS、甲苯胺蓝、Van Gieson染色和Mallory染色,观察骨关节组织的形态学变化,并对几种染色方法进行比较。结果 HE染色显示关节一般组织形态结构,可见模型组关节软骨和软骨下骨发生骨性关节炎病理变化;番红-固绿染色法中软骨和软骨下骨的界限（黏合线）以及潮线显示清晰,软骨基质中糖胺聚糖含量减少,纤维成分增多;AB-PAS染色显示骨关节炎软骨基质糖胺聚糖尤其是酸性糖胺聚糖含量减少;甲苯胺蓝染色显示骨关节炎软骨的酸性糖胺聚糖减少;Van Gieson染色和Mallory染色可显示骨关节组织中的胶原纤维,但组织结构界限不够清晰。结论在骨性关节炎的组织形态学研究中,通过常规HE染色,结合番红-固绿染色法和AB-PAS染色法,能较客观全面地获得关节组织形态学相关信息。     

冻融联合灌注法优化大鼠肾脏脱细胞支架的制备

本研究旨在探索优化肾脏脱细胞支架的制备方法,为肾脏组织工程及肾脏体外病理、毒理研究提供实验基础。取大鼠肾脏灌注PBS作为对照组 (Control组),在不同流速下分别以十二烷基磺酸钠 (Sodium dodecyl sulfate,SDS) 灌注 (S组),Triton X-100联合SDS灌注 (TS组),反复冻融后Triton X-100联合SDS灌注(FTS组),制备肾脏脱细胞支架,并测定其流体分布及脉管阻力。HE染色、DAPI染色、DNA定量检测脱细胞支架脱细胞程度,Masson染色、PAS染色、免疫组织化学染色检测脱细胞支架主要成分的保留和结构的完整,扫描电镜检测支架的超微结构,MTT法检测支架的细胞毒性,ELISA检测支架中生长因子的含量。结果显示,FTS组脱细胞用时较S组、TS组少,10 mL/min组支架脉管阻力较低,S组、TS组、FTS组流体分布与Control组存在差异。HE染色和DAPI染色显示各组支架未见细胞成分残留,DNA含量＜50 ng/mg。Masson染色和PAS染色可见细胞外网状胶原及多糖,免疫组织化学染色见Ⅰ型胶原 (CollagenⅠ)、Ⅳ型胶原蛋白 (Collagen Ⅳ)、纤维连接蛋白 (Fibronectin)、层粘连蛋白 (Laminin) 表达。扫描电镜见支架呈蜂窝状结构。MTT法检测支架细胞毒性分级在0–1级之间。ELISA检测提示FTS组VEGF、EGF、IGF-1、PDGF含量明显高于S组和TS组。综上,联合冻融和灌注法能够制备更为理想且有效的大鼠肾脏整器官脱细胞支架,为肾脏组织工程及肾脏体外病理、毒理学研究奠定基础。     

淫羊藿总黄酮通过TGF-β1/Smad3信号通路改善自然衰老大鼠肾脏组织纤维化

为探讨淫羊藿总黄酮(TFE)对自然衰老大鼠肾脏纤维化的影响及其可能的作用机制,将18月龄SD大鼠随机分为自然衰老组、淫羊藿总黄酮低(10 mg/kg)、高(40 mg/kg)剂量组,另取2月龄SD大鼠作为青年对照组,给药4个月。通过HE染色和Masson染色观察肾脏组织形态及胶原纤维含量,生化酶法检测肾脏组织SOD活力和MDA含量,免疫组化法检测αSMA的蛋白表达水平,实时定量PCR和Western blot技术检测TGF-β1和Smad3的mRNA及蛋白表达水平。结果显示,TFE(低、高剂量)可改善衰老SD大鼠肾脏组织形态,减少胶原纤维,提升肾脏组织SOD活力(P<0.05),降低MDA含量(P<0.05),并降低肾脏组织中αSMA蛋白表达(P<0.05),同时显著下调TGF-β1及Smad3的mRNA和蛋白表达(P<0.05)。淫羊藿总黄酮可改善自然衰老大鼠肾脏纤维化,其机制可能与其抑制TGFβ1/Smad3信号通路传导有关。     

SARS-CoV N蛋白与Smad3相互作用并调节TGF-β信号通路的形态学初步研究

目的应用病理学实验技术,初步描述SARS-CoV N蛋白与TGF-β信号通路中Smad3蛋白之间相互作用的情况,及其对TGF-β信号通路下游转录产物生成的影响。方法对比观察SARS-CoV感染的恒河猴肺组织及正常猴肺组织的形态学特征,应用免疫组织化学染色法,观察猴肺组织中转化生长因子-β（transforming growth factor-β,TGF-β）及1型纤维蛋白溶酶原活化抑制剂（plasminogen activator inhibitor-1,PAI-1）的肺组织表达;SARS-CoVNucleocapsid蛋白与Smad3双染观察二者在肺组织中的共定位情况;应用Masson染色法比较感染与正常猴肺组织中胶原含量的差异,结合分子生物学检测肺组织中TGF-β、PAI-1及Ⅰ型胶原α2链（α2 chain of typeⅠcollagen,COLlA2）的表达,探讨N蛋白影响TGF-β信号通路的影响。结果PCR结果显示TGF-β、PAI-1及COL1A2在SARS-CoV感染的猴肺组织中的表达量较正常猴肺有明显增高,免疫组织化学染色结果提示,在病毒感染的猴肺组织内TGF-β、PAI-1染色呈现强阳性,同时SARS-CoV Nucleocapsid蛋白与Smad3双染结果显示,二者在感染的猴肺组织中存在明显的共定位现象。Masson染色结果提示,病毒感染的猴肺组织胶原含量较正常猴肺组织显著增加。结论我们的研究结果为说明SARS-CoV Nucleocapsid蛋白参与调节TGF-β信号通路,促进肺组织纤维化提供了有力的形态学实验证明,为进一步理解SARS-CoV感染引起肺组织纤维化的机制提供了的较为明确体内实验数据。     

慢性肾衰竭大鼠模型的建立

目的 建立两种慢性肾衰竭大鼠模型,观察瘦素蛋白在大鼠组织、器官中的表达.方法 建立两种慢性肾衰竭CRF动物模型:(1)大鼠肾大部分切除诱发肾衰(Platt法).(2)腺嘌呤诱发大鼠慢性肾衰竭的动物模型(Yokozawa法).分别测定血清中血尿素氮(BUN),血肌酐(Scr)Ca2+、P5+等含量.取肾脏组织,HE染色,行免疫荧光,检测瘦素蛋白在两种慢性肾衰竭大鼠模型中的表达情况.结果 模型组大鼠血清中血尿素氮(BUN),血肌酐(Scr)等含量明显升高,免疫荧光检测显示两种模型大鼠肾脏组织瘦素蛋白的表达.结论 成功建立两种慢性肾衰竭CRF动物模型,显示不同模型组织部位的瘦素蛋白的表达.为进一步探讨瘦素蛋白在动物体内的生物学作用提供实验基础.     

大鼠咬合创伤牙周膜中MCP-1、ICAM-1的表达变化

目的:研究咬合创伤大鼠牙周组织中MCP-1、ICAM-1的表达情况。方法:12周龄雄性SD大鼠24只,随机分为4组(1个正常对照组和3个实验组),每组6只。正常对照组不作任何处理,实验组通过在大鼠左上颌第一磨牙颌面粘接树脂并内置不锈钢丝形成高出颌面0.6-0.8 mm的树脂层以建立同侧下颌咬合创伤实验动物模型,分别于建模后3、5、7 d处死各组大鼠,分离大鼠下颌组织,运用HE、Masson染色观察咬合创伤牙周组织形态变化,同时用免疫组织化学染色法检测MCP-1和ICAM-1的表达变化。结果:HE染色显示,正常组牙周膜纤维排列整齐,牙骨质表面较为平整,牙槽骨结构致密。实验组牙周膜纤维排列紊乱,牙周膜血管水肿充血、间隙改变,牙槽骨和牙骨质表面不平整,出现骨吸收。Masson染色显示,正常组牙周组织未见异常表现;实验组牙周膜纤维排列紊乱,可见水解断裂,局部有血流障碍和血管破裂。免疫组织化学显示,各实验组MCP-1和ICAM-1的表达变化均较正常对照组增多,差异有显著性(P0.05)。其中7 d组表达水平最高,与其他2组相比有统计学意义(P0.05)。结论:咬合创伤可引起大鼠牙周组织形态变化,MCP-1、ICAM-1的表达随时间呈现递增的趋势。     

胶原蛋白/BMP复合材料的制备和成骨性能研究

以胶原膜(含87.5 mg I型胶原蛋白)为载体, 复合3.5 mg rhBMP-2(人基因重组骨形成蛋白-2), 制备胶原蛋白/BMP复合材料。复合材料首先在兔背阔肌中埋置, 预构新生骨组织, 并采用ALP染色、Von Kossa染色和HE染色等观察复合材料的成骨过程和组织形态。然后将形成的新骨组织游离移植修复自体下颌骨体部洞穿性缺损; 并设以胶原为载体的rhBMP-2复合骨修复材料直接修复为对照组, 骨缺损不修复组为空白组。采用X线、抗压强度、硬组织切片、四环素荧光染色、骨形态计量检查, 观察复合材料修复骨缺损的质量和效果。结果表明, 胶原蛋白/BMP复合材料在兔背阔肌中4～6周成骨, 胶原材料于3～5周降解; 成骨过程为是以软骨成骨为主的方式, 新骨形态为编织骨, 可见明显的微血管分布; 游离移植修复自体下颌骨缺损, 6周缺损区为骨性愈合, 与对照组在抗压强度(P = 0.041)、新骨量(P = 0.034)均有显著性差异。胶原蛋白/BMP复合材料在骨骼肌中形成的新生骨组织可作为供骨修复一定范围的骨缺损。     

胶原蛋白/BMP复合材料的制备和成骨性能研究

以胶原膜(含87.5 mg I型胶原蛋白)为载体, 复合3.5 mg rhBMP-2(人基因重组骨形成蛋白-2), 制备胶原蛋白/BMP复合材料。复合材料首先在兔背阔肌中埋置, 预构新生骨组织, 并采用ALP染色、Von Kossa染色和HE染色等观察复合材料的成骨过程和组织形态。然后将形成的新骨组织游离移植修复自体下颌骨体部洞穿性缺损; 并设以胶原为载体的rhBMP-2复合骨修复材料直接修复为对照组, 骨缺损不修复组为空白组。采用X线、抗压强度、硬组织切片、四环素荧光染色、骨形态计量检查, 观察复合材料修复骨缺损的质量和效果。结果表明, 胶原蛋白/BMP复合材料在兔背阔肌中4～6周成骨, 胶原材料于3～5周降解; 成骨过程为是以软骨成骨为主的方式, 新骨形态为编织骨, 可见明显的微血管分布; 游离移植修复自体下颌骨缺损, 6周缺损区为骨性愈合, 与对照组在抗压强度(P = 0.041)、新骨量(P = 0.034)均有显著性差异。胶原蛋白/BMP复合材料在骨骼肌中形成的新生骨组织可作为供骨修复一定范围的骨缺损。     

巨噬细胞在小鼠肾纤维化恢复期的研究

目的:观察巨噬细胞在小鼠肾纤维化进展期和恢复期的作用。方法:采用单侧输尿管结扎(UUO)肾纤维化模型和输尿管再通模型(RUUO)进行试验研究;用Masson染色和HE染色观察肾脏纤维化程度和炎症变化趋势;流式分析肾脏中巨噬细胞细胞群的比例变化。结果:Masson染色显示肾脏纤维化程度在梗阻解除后胶原沉积面积减轻从80%降到46%,差异有统计学意义(P0.05)。HE染色显示梗阻解除后肾间质炎症减轻,且有新生小管形成。流式结果显示梗阻解除后巨噬细胞细胞群比例由19%降到2.6%,差异有统计学意义(P0.05)。结论:巨噬细胞可能在肾纤维化恢复期发挥一定作用。     

Increased arginase activity and endothelial dysfunction in human inflammatory bowel disease

Nitric oxide (.NO) generation from conversion of l-arginine to citrulline by nitric oxide synthase isoforms plays a critical role in vascular homeostasis. Loss of .NO is linked to vascular pathophysiology and is decreased in chronically inflamed gut blood vessels in inflammatory bowel disease (IBD; Crohn's disease and ulcerative colitis). Mechanisms underlying decreased .NO production in IBD gut microvessels are not fully characterized. Loss of .NO generation may result from increased arginase (AR) activity, which enzymatically competes with nitric oxide synthase for the common substrate l-arginine. We characterized AR expression in IBD microvessels and endothelial cells and its contribution to decreased .NO production. AR expression was assessed in resected gut tissues and human intestinal microvascular endothelial cells (HIMEC). AR expression significantly increased in both ulcerative colitis and Crohn's disease microvessels and submucosal tissues compared with normal. TNF-alpha/lipopolysaccharide increased AR activity, mRNA and protein expression in HIMEC in a time-dependent fashion. RhoA/ROCK pathway, a negative regulator of .NO generation in endothelial cells, was examined. The RhoA inhibitor C3 exoenzyme and the ROCK inhibitor Y-27632 both attenuated TNF-alpha/lipopolysaccharide-induced MAPK activation and blocked AR expression in HIMEC. A significantly higher AR activity and increased RhoA activity were observed in IBD submucosal tissues surrounding microvessels compared with normal control gut tissue. Functionally, inhibition of AR activity decreased leukocyte binding to HIMEC in an adhesion assay. Loss of .NO production in IBD microvessels is linked to enhanced levels of AR in intestinal endothelial cells exposed to chronic inflammation in vivo.     

益气化瘀化痰方对肺纤维化大鼠的干预作用及其机制*

目的: 探讨益气化瘀化痰方对由博来霉素(BLM)诱导的大鼠肺纤维化的影响及其可能的机制。方法: 采用气管内一次性注射盐酸博来霉素5 mg/kg诱导制备肺纤维化大鼠模型。将60只SPF级SD大鼠随机分为正常对照组、模型对照组、阳性对照组、益气化瘀化痰方低浓度组(低浓度组)、益气化瘀化痰方中浓度组(中浓度组)和益气化瘀化痰方高浓度组(高浓度组),每组10只。各中药治疗组在造模4周后分别采用益气化瘀化痰方低、中、高浓度制剂(3.6 ml/(kg·d))灌胃,阳性对照组给予醋酸氢化可的松(3.6ml/(kg·d))灌胃,正常对照组和模型对照组给予同等体积的生理盐水(3.6 ml/(kg·d))灌胃,每日1次。12周后处死各组大鼠,取大鼠肺组织,称量肺质量并计算肺系数,HE染色观察组织形态学改变,Masson染色观察胶原纤维沉积情况;蛋白免疫印迹(Western blot)检测各组大鼠肺组织中转化生长因子-β1(TGF-β1)、锌指蛋白转录因子(Snail1)、E-钙黏蛋白(E-cadherin)、纤连蛋白(Fibronectin)等蛋白表达水平;实时荧光定量PCR(qRT-PCR)检测组织TGF-β1、Snail1、E-cadherin、Fibronectin等mRNA表达水平。结果: ①与模型组比较,各药物干预组肺系数均有降低(P＜0.05);②与空白组比较,模型组肺纤维化明显;各药物干预组与模型组比较,各治疗组肺间质胶原沉积减少,肺纤维化程度减轻;③治疗各组与模型组比较,各治疗组大鼠肺组织TGF-β1、Snail1、Fibronectin蛋白表达均明显减少(P＜0.05),E-cadherin蛋白表达明显升高(P＜0.05);④低、高浓度组与中浓度组比较,尤以中浓度组表达结果最显著(P＜0.05);⑤中药治疗各组与阳性组比较,表达结果优于阳性组(P＜0.05);mRNA表达与蛋白表达趋势一致。结论: 益气化瘀化痰方能显著治疗博来霉素所致的大鼠肺纤维化,其机理可能是通过下调TGF-β1调控TGF-β/Snail信号通路表达抑制大鼠的上皮细胞-间充质转化,而改善大鼠肺纤维化。     

Tissue Explants Used for Enhancement of Herpesvirus Isolations

Herpesvirus recovery from normal and experimentally infected simian tissues prepared into 10 to 20% homogenates was compared with that obtained by explanting these same tissues. The explant procedure was by far the more sensitive of the two methods. As a result, 8 herpesvirus isolations were obtained by conventional virus isolation procedures in contrast to 74 isolates when explants were made from these same tissue sources. There is also evidence to suggest that the explant method is more sensitive than the use of tissue homogenates for recovery of other viruses as well.     

特殊染色及免疫组化双重染色在恒河猴糖尿病模型中的应用

目的在链脲佐菌素（STZ）诱导恒河猴糖尿病动物模型基础上,用特殊染色、免疫组化双染方法显示胰腺等组织中的特征性病变。方法健康恒河猴5只,小剂量（30mg／kg）多次静脉注射STZ,濒死状态时将动物安乐死。取胰腺、心脏、肾脏、脾脏、肝脏、眼球、脑等器官制成石蜡切片,用HE、PAS、Masson、天狼星红和甲苯胺蓝等方法进行特殊染色,用免疫组化双重染色法同时显示胰岛A、B细胞。结果模型动物的胰岛萎缩,数量减少。Masson染色见外分泌部间质内纤维增生。局灶性慢性肾炎,病变区肾间质纤维组织增生,部分肾小球及肾小管萎缩。天狼星红染色见脾脏中央动脉管壁增厚。免疫组化双染见胰岛A胰高血糖素表达增多,胞浆呈棕褐色。B胰岛素表达减少,胞浆粉红色。结论HE染色结合特殊染色和免疫组化双重染色可较好地对STZ诱导糖尿病动物模型进行组织学评价。