目的：比较经不同固定液处理的小鼠肝脏石蜡切片H E染色效果，优化实验方法。方法用3种不同固定液（10%中性福尔马林、4%多聚甲醛、A F固定液）处理小鼠肝脏组织，经常规脱水、透明、浸蜡、包埋、切片，H E染色，观察比较染色效果。结果10%福尔马林、4%多聚甲醛固定的肝组织结构完整、清晰，细胞界限清楚，染色鲜艳；A F固定液固定的肝组织结构不完整，界限不清，胞浆中出现大量空泡。结论10%中性福尔马林和4%多聚甲醛是制作小鼠肝脏石蜡切片HE染色标本的理想固定液。

Purpose: To compare of mouse liver paraffin section and HE staining effect treatment with different fixative, optimization of experimental method. Methods: Mouse livers were immersed in three different fixatives (10% buffered formalin, 4% polyformaldehyde, and AF fluid) respectively. The tissue were dehydrated, transparentized and immersed in paraffin as routine procedure. The paraffin embedded liver tissue were sectioned and stained with Hematoxylin and Eosin, liver sections were compared under the light microscope. Results: The tissues fixed with 10% buffered formalin and 4% polyformaldehyde showed complete and clear structure, and the hepatocytes wel stained. The structure of liver tissues fixed with AF fluid was not complete, il-defined, and with large vacuoles in cytoplasm.Conclusion: 10% buffered formalin and 4% polyformaldehyde fixative is the most effective fixative for mouse liver HE staining.

目的：分析10%福尔马林、纯丙酮、95%乙醇、甲醇4种固定液对脑组织冷冻切片H-E染色质量的影响，筛选出一种脑组织冷冻切片的最佳固定液。方法收集手术室送检新鲜脑组织标本，经常规冷冻切片后放入不同固定液，进行H-E染色，观察脑组织冷冻切片细胞收缩程度和染色质量。结果甲醇组细胞收缩不明显，相对于其他组比较差异有统计学意义（P＜0.05）；甲醇组组织结构清晰，核浆染色鲜艳，对比度好，相对于其他组差异性显著（P＜0.001）。结论甲醇固定脑组织染色效果好，使用方便，满足术中冷冻切片的快速、准确诊断，是脑组织冷冻切片的理想固定液。

Objective:To screen the best fixative on brain tissue frozen section,we analysed that 4 fixatives including 10% formalin,pure acetone,95% ethanol,and methanol impact on the quality of H&E staining quality.Methods:Fresh brain tissue specimens were colected,and we made frozen sections in the different fixatives,and observed brain tissue shrinkage and dying quality with H&E staining. Results:Cel shrinkage of methanol group was significantly less than other groups (P<0.05); For methanol group,organization structure is more clear,the color is more bright and nuclear plasma contrast is higher,relative to other groups (P<0.001).Conclusion: methanol fixation method is effective on staining,easy to use,and contributing to the rapid and accurate diagnosis of intraoperative frozen section. Methanol is an ideal fixative for brain tissue frozen section.

目的 探讨大鼠脑挫伤后损伤侧与非损伤侧脑组织病理变化及其意义. 方法 选择健康成年Wistar大鼠100只,按随机数字表法分为假手术组和挫伤组,各组又分为1,3,6,12,24 h5个时相点,每个时相点10只大鼠.各时相点取脑组织分别行挫伤处及非挫伤处病理形态学观察,IgG免疫组化染色半定量分析检测血脑屏障(blood brain barrier,BBB)通透性. 结果 假手术组双侧脑组织结构未见异常,IgG染色阴性.挫伤组损伤侧伤后1h即出现以BBB破坏为特征的血管源性水肿,3h出现细胞内水肿,两种水肿均随时间延长逐渐加重.挫伤后12 h后开始出现组织坏死、炎性细胞浸润及小胶质细胞增生.伤后1 h IgG开始呈阳性,6h最强,维持高水平至24 h.而挫伤组非损伤侧伤后1h结构未见异常,IgG染色阴性.3h后出现细胞内水肿,IgG染色弱阳性.6h后细胞内水肿加重,同时出现血管源性水肿,IgG染色呈强阳性.12～24 h细胞内水肿仍继续加重,而血管源性水肿有缓解趋势,IgG染色减弱,并见小胶质细胞增生. 结论 脑挫伤后损伤侧与非损伤侧病理改变有差异,可为脑挫伤治疗方法选择提供依据.

Objective To investigate the pathological changes of brain tissues from contused and contralateral non-contused sides and their significance.Methods A total of 100 healthy adult Wistar rats were assigned to brain contusion group and sham operation group according to the random number table.Ten rats in each group were sacrificed at 1,3,6,12,and 24 hours respectively.Brain samples were collected to perform pathophysiological analysis of brain tissues and test blood brain barrier (BBB)permeability by semiquantitative immunohistochemical staining of IgG.Results There was no damage to the bilateral brain tissues in sham operation group and IgG stain was negative.In brain contusion group,angioedema characterized by the breakdown of BBB was seen in the contused side at 1 hour followed by cellular edema at 3 hours,with aggravation of both over time.Moreover,tissue necrosis,inflammatory cell infiltration,and microglia proliferation emerged at 12 hours.Besides,IgG-positive staining was seen at 1

采用单纯胰蛋白酶消化法分离,差速贴壁法纯化新生大鼠心肌细胞。光学显微镜观察心肌细胞基本形态特征,台盼蓝染色检测成活率,肌钙蛋白Ⅰ、α-横纹肌肌动蛋白抗体鉴定心肌细胞。结果显示培养24 h后心肌细胞大部分已贴壁,并出现自发搏动现象,48 h后逐渐伸出伪足相互接触交织成网,72 h后逐渐形成细胞簇并出现同步搏动;台盼蓝染色法检测细胞成活率达90%以上;肌钙蛋白Ⅰ及α-横纹肌肌动蛋白免疫荧光染色法鉴定心肌细胞纯度达98%以上。

Neonatal rat myocytes were digested with typsin and purified by technique of differential anchoring. The cardiomyocytes cultured were observed under light microscope, stained with trypan blue to count the living cells and identified with cTnI andα-Sarcomeric actin. The results showed most of cardiomyocytes adherented to culture plates after 24 hours of culture and a part of cells were jumping synchronously,48 hours later cells gradually extended pseudopodia,contacted with each other and interwovened into a network. After 72 hours of culture, the cells gradu-ally formed cell clusters and were beating synchronously. Trypan blue staining showed the survival rate of cardio-myocytes was more than 90%. cTnI andα-Sarcomeric actin immunofluorescence staining showed that the purity of cardiomyocytes cultured was more than 98%.

目的研究溴酚蓝对大鼠胶质瘤模型的染色作用，为手术精确切除人脑胶质瘤提供实验基础。方法建立Fischer 344大鼠胶质瘤模型，MRI检查证实。静脉注射不同剂量（30～360mg/kg）溴酚蓝，15min后处死取脑，检查溴酚蓝对脑肿瘤的染色情况；并注射相同剂量（180mg/kg）的溴酚蓝，在不同时间处死取脑，检查溴酚蓝对脑肿瘤的染色情况。对无瘤实验大鼠注射不同剂量溴酚蓝，观察其毒性反应。结果溴酚蓝对大鼠胶质瘤有明显的染色作用，其染色与周围脑组织有清晰的界限，染色时间出现在注射后10min～6h。30d未观察到溴酚蓝的任何毒性反应。结论溴酚蓝在不同时间点能够清晰染色大鼠胶质瘤，有可能作为人脑胶质瘤的术中染色剂来区别肿瘤组织和正常脑组织。

Objective To investigate the uptake of bromophenol blue (BPB) in experimental rat glioma. Methods Glioma model was established in Fischer 344 rats and confirmed by MRI. Different doses (30~360mg/kg) of BPB were administrated by intravenous injection in glioma- bearing rats. Fifteen minutes after BPB injection, the animals were sacrificed and their brains were removed for monitoring tumor staining. In a subsequent study, 180 mg/kg BPB was injected and the animals were sacrificed at several time points to monitor tumor staining over time. Also, we conducted a toxicity study with intravenous injections of different dose BPB in nontumor- bearing Fischer 344 rats. Results The BPB staining was clearly visible and mainly localized in the tumor. In an additional experiment, we found that tumor staining persisted from 10min to 6h after BPB injection. No adverse effects were observed in animals during the 30d- observation period. Conclusion BPB can visualize experimental rat glioma at different