Comparison of Hematoxylin-eosin Staining and Methyl Violet Staining for Displaying Ghost Cells

Purpose:.To compare the merits and limitations of hematoxylin-eosin(HE) and methyl violet staining for displaying ghost cells from vitreous or aqueous humor.Methods:.A specimen containing ghost cells was adjusted to five different concentrations:(12×104,.10×104,.8×104, 6×104and 4×104cells / ml) and subjected to smearing and methyl violet and HE staining..The staining results were observed by light microscopy.Results: The ghost cells were readily observed at a cell density of > 8×104cells / ml with methyl violet staining,.but only a few cells were occasionally seen at lower cell densities..In contrast,.ghost cells were seen at all cell densities with HE staining.Conclusion: Methyl violet staining is more rapid and simpler for the identification of ghost cells, but its staining color more readily fades, the slides cannot be stored, and it is only effective at a cell density of > 8 ×104cells / ml. In contrast,.HE staining is more time-consuming but it can display cell morphology and distinguish cell components more explicitly and slides can be permanently stored. HE staining has advantages over methyl violet staining in detecting the ghost cells when the concentration is < 8×104cells / ml.

DNA extraction from archived hematoxylin and eosin-stained tissue slides for downstream molecular analysis

BACKGROUND Histopathologically stained archived tissue slides are stored in hospital archives for years to decades.They are the largest available source of biological materials and are a potentially useful resource that can be used for retrospective epidemiological studies.DNA recovered from the slides can be used for several downstream molecular processes including polymerase chain reaction,single nucleotide polymorphism analysis,and whole genome sequencing.The DNA from these slides can be utilized to compare gene signatures of normal and diseased tissues.However,extraction of high-quality DNA from archived stained hematoxylin and eosin(H&E)slides remains challenging.AIM To standardize a new protocol for extracting DNA from archived H&E-stained tissue slides for further molecular assays.METHODS A total of 100 archived H&E-stained cancer slides were subjected to a total of five methods of DNA extraction.Methods were varied in the deparaffinization step,tissue rehydration,duration of lysis,and presence or absence of proteinase K.The extracted DNA was quantified using a NanoDrop spectrophometer and the quality was analyzed by agarose gel electrophoresis.Then each sample was subjected to polymerase chain reaction(PCR)to amplify the internal control gene GAPDH,thereby confirming the DNA intactness,which could be further utilized for other downstream applications.RESULTS Of the five different methods tested,the third method wherein xylene was used for tissue deparaffinization followed by 72 h of digestion and without proteinase K inactivation yielded the highest amount of DNA with good purity.The yield was significantly higher when compared to other methods.In addition,90%of the extracted DNA showed amplifiable GAPDH gene.CONCLUSION Here we present a step-by-step,cost-effective,and reproducible protocol for the extraction of PCR-friendly DNA from archived H&E-stained cancer tissue slides that can be used for further downstream molecular applications.

A comparing study of quantitative staining techniques for retinal neovascularization in a mouse model of oxygen-induced retinopathy

AIM: To explore an efficient, practical and objective quantitative method to evaluate the retinal neovascularization in mouse model of oxygen induced retinopathy (OIR). METHODS: Thirty C57BL/6J mice were explored in OIR model procedure. Eyes were removed for different staining methods including: (1) HE staining; (2) immunohistochemistry with Griffonia Simplicifolia Lectin (GSL); (3) Immunofluorescence with FITC labeled CD31 antibody; (4) Two-step immunofluorescence with purified-CD31 antibody; (5) FITC-Dextran perfusion combined with two-step purified-CD31immunofluorescence. Images of the retinal vasculature were analyzed by imaging software. ' RESULTS: GSL immunohistochemistry could clearly demonstrate the deep and superficial capillary beds. FITC labeled CD31 Immunofluorescence was blurring with high fluorescence background which was hard to distinguish retinal neovascularization in some area. Excellent detail of neovascularization and preexistent retinal vessels was provided in two-step Purified-CD31 immunofluorescence group. CONCLUSION: GSL immunohistochemistry can clearly demonstrate neovascularization tufts in deep and superficial capillary beds. Immunofluorescence of specific antigen CD31 on vascular endothelium can selectively label the neovascularization of mouse retina. When combined with computer analysis software, it is an effective and objective quantitative method to evaluate the retinal neovascularization in OIR mouse model.

Digitally reinforced hematoxylin-eosin polarization technique in diagnosis of rectal amyloidosis

AIM:To investigate the efficacy of the digitally reinforced hematoxylin-eosin polarization(DRHEP)technique for detection of amyloidosis in rectal biopsies.METHODS:One hundred hematoxylin-eosin(HE)stained rectal biopsies with Congo-red(CR)-positive amyloid depositions and 50 control cases with CRnegative amyloid-mimicking areas were scanned blinded to the CR results for amyloid depositions under both bright and polarized light,and digitally photographed using the DRHEP technique,to accentuate the faint birefringence observed in HE slides under polarization.The results of DRHEP and HE evaluation were statistically correlated with CR polarization results with respect to presence and localization of amyloid deposits as well as amyloid types.RESULTS:Amyloid deposits showed yellowish-green birefringence by DRHEP,which allowed identification of amyloidosis in 41 HE-unsuspected cases(P=0.016),31 of which only had vascular deposits.True positivity was higher,and false negativity and positivity were lower by DRHEP,compared to evaluation by HE(69%,31%,and 0.8%vs 33%,67%,and 33%,respectively;P<0.0001).The sensitivity,specificity,accuracy,and positive and negative predictive values for DRHEP were69%,98%,78.6%,98.5%,and 61.25%,respectively.Reasons for DRHEP false negativity were presence of extensive background birefringence in 12 cases,absence of CR birefringent vessel in 3 cases,and missing of the tiny deposits in 9 cases,which could be improved by experience,especially in the latter case.No correlation was found between age,gender,sites of deposits,or amyloid types.CONCLUSION:The DRHEP technique improves diagnostic accuracy when used as an adjunct or a prior step to CR staining,especially for cases with limited tissues for further analysis.

Diagnostic value of endothelial markers and HHV-8 staining in gastrointestinal Kaposi sarcoma and its difference in endoscopic tumor staging

AIM: To clarify the diagnostic values of hematoxylin and eosin (HE), D2-40, CD31, CD34, and HHV-8 immunohistochemical (IHC) staining in gastrointestinal Kaposi's sarcoma (GI-KS) in relation to endoscopic tumor staging. METHODS: Biopsy samples (n = 133) from 41 human immunodeficiency virus-infected patients were reviewed. GI-KS was defined as histologically negative for other GI diseases and as a positive clinical response to KS therapy. The receiver operating characteristic area under the curve (ROC-AUC) was compared in relation to lesion size, GI location, and macroscopic appearances on endoscopy. RESULTS: GI-KS was confirmed in 84 lesions (81.6%). Other endoscopic findings were polyps (n = 9), inflammation (n = 4), malignant lymphoma (n = 4), and condyloma (n = 2), which mimicked GI-KS on endoscopy. ROC-AUC of HE, D2-40, blood vessel markers, and HHV-8 showed results of 0.83, 0.89, 0.80, and 0.82, respectively. For IHC staining, the ROC-AUC of D2-40 was significantly higher (P < 0.05) than that of HE staining only. In the analysis of endoscopic appearance, the ROC-AUC of HE and IHC showed a tendency toward an increase in tumor staging (e.g. , small to large, patches, and polypoid to SMT appearance). D2-40 was significantly (P < 0.05) advantageous in the upper GI tract and for polypoid appearance compared with HE staining. CONCLUSION: The diagnostic value of endothelial markers and HHV-8 staining was found to be high, and its accuracy tended to increase with endoscopic tumor staging. D2-40 will be useful for complementing HE staining in the diagnosis of GI-KS, especially in the upper GI tract and for polypoid appearance.

Role of Immunostaining in Detecting Extra-Pattern and Subtle Lymphomatous Infiltration in Bone Marrow Biopsies of NHL Patients

Introduction: Immunohistochemistry (IHC) enables the examination of a greater number of trephine biopsy levels and is helpful in determining additional scattered malignant cells. The aim of this study is to detect extra-pattern and subtle lymphomatous infiltration in bone marrow biopsies using CD20 and CD3 immunostaining. Patients and Methods: This study was conducted on 100 newly diagnosed Non Hodgkin Lymphoma (NHL) patients. Their bone marrow trephine biopsies were assessed on routine histology [Hematoxylin and Eosin (H & E)], and were further subjected to IHC using CD20 and CD3. Results: Pattern of involvement by H & E was highlighted by IHC. It showed additional interstitial pattern in 9 cases, parasinusoidal streaks in one case and highlighted a patchy pattern in another case with interstitial involvement on H & E. IHC also detected subtle infiltrations on additional 5.5% cases compared with histology alone. It helped in differentiating reactive (12 cases) and malignant lymphoid infiltration (33 cases). Conclusion: CD20 and CD3 immunostaining performed routinely on bone marrow trephine biopsies has the ability to reveal extra-pattern of infiltration and improve detection of subtle lymphoid involvement. A combined procedure identifying several distinctive features, in particular histotopography and IHC, provides a promising way of discriminating reactive from neoplastic lymphoid infiltrates in bone marrow trephine biopsies.

不同固定方法对豚鼠眼球后极部的固定效果比较

目的:为解决豚鼠眼球组织切片制备过程中所存在的视网膜脱片问题,采用不同固定方法,优化固定效果。方法:2周龄正常豚鼠(75只)随机分为5个大组(A-E组),15个小组,每小组5只。A组(1-3小组)眼球分别置于FAS、Davidson固定液1(D1)和Davidson固定液2(D2)中固定24 h;B组(4-6小组)眼球在固定液中固定1 h后剪切角膜,再固定2 h;C组(7-9小组)眼球在固定液中固定1 h后沿视神经方向将眼球分为左右两半,再固定2 h;D组(10-12小组)眼球在固定液中固定3 h后将眼球分为左右两半;E组(13-15小组)眼球在固定液中固定3 h后剪切角膜。经苏木精-伊红(HE)染色比较各个小组眼球后极部固定效果。结果:形态观察表明1-6小组、11-15小组眼球表面光滑圆润,色泽透明,7-10小组眼球凹陷皱缩变形。HE染色表明大部分组别眼球后极部组织切片卷曲缠绕,视网膜脱离;1、5、6、14、15小组切片结构规整,其中14小组形态最佳,视网膜、脉络膜、巩膜连接紧密,组织结构清晰,细胞排列规整。结论:采用D1固定液固定3 h后剪切角膜的固定效果最为理想,适用于豚鼠眼球后极部相关组织研究。

病原菌血流感染对阿尔茨海默病小鼠记忆力和海马锥体细胞的影响

目的 探讨病原菌血流感染对阿尔茨海默病(AD)小鼠记忆力和海马锥体细胞的影响。方法 50只C57BL/6J雄性小鼠腹壁皮下注射D-半乳糖液[200 mg/(kg·d)]连续8周构建AD模型并于造模期间每周观测小鼠的一般状态及体质量;造模结束后均分为生理盐水(对照)组、大肠埃希菌组、铜绿假单胞菌组、肺炎克雷伯杆菌组、金黄色葡萄球菌组及白念珠菌组,对照组小鼠尾静脉注射生理盐水,其余病原菌感染组小鼠尾静脉注射低(1.5×107CFU/L)、中(1.5×108CFU/L)及高(1.5×109CFU/L)浓度的对应病原菌菌液,观察14 d,分别于注射后第1、2、3天及第1、2周测定各组小鼠的体质量及体温;观察结束后,于第15天采用Morris水迷宫实验检测各组小鼠逃逸潜伏期、游泳速度及穿越平台次数,水迷宫实验共进行6 d;实验结束后处死各组小鼠,取海马组织,采用苏木精-伊红(HE)染色观察海马组织学的改变。结果 小鼠于第8周体质量开始下降,且精神状况较造模初期明显下降;部分病原菌感染组小鼠逃避潜伏期较对照组延长(P<0.05),第5天肺炎克雷伯菌高浓度组小鼠水迷宫实验游泳速度较对照组下降(P<0.05);对照组小鼠海马CA1和CA3区神经细胞及皮层未见明显异常,白念珠菌组小鼠海马组织中发现少量凋亡细胞,肺炎克雷伯菌组、金黄色葡萄球菌组小鼠海马神经细胞轻度排列紊乱。结论 AD患者来源的病原菌经尾静脉血流感染可导致AD小鼠记忆力下降及海马锥体细胞的形态改变。

二次脱钙法在临床病理骨组织HE制片中的应用研究

目的 探讨二次脱钙法在临床病理骨组织苏木精-伊红(HE)制片中的应用价值。方法 收集2022年6-12月该院病理科脱钙不充分、骨组织块或含钙化组织的蜡块100块,制成病理切片,根据脱钙方法的不同将病理切片分为对照组和研究组,对照组采用常规脱钙法,研究组采用二次脱钙法,分析2种方法制片效果的差异。结果 研究组病理切片HE制片结构完整性、HE染色满意度、切片优良率均明显高于对照组,差异均有统计学意义(P<0.05)。结论 二次脱钙法在临床病理骨组织HE制片中具有较高的应用价值。

胃肠活检组织常规染色不良原因分析

目的探讨胃肠活检组织在常规苏木精-伊红染色(HE)中出现伊红着色不均的原因分析。方法参照实用现代病理学技术的制片流程,对可能存在问题的组别一一排查。收集2023年8月至2024年1月中山市人民医院存档的120例胃肠活检石蜡标本切片,在染色不同时期进行上机HE染色,对比分析不同染色时期下的染色结果。结果排查后发现问题可能存在HE染色步骤组中,乙醇浓度下降对伊红着色有影响。设置3批片,在不同染色时期分别上机染色。镜下观察前、中、后3个染色时期中,伊红着色不均率分别为0%(0/40)、17.5%(7/40)、72.5%(29/40)。在不同染色时期中伊红着色不均率差异有统计学意义(χ^(2)=78.603,P<0.01)。结论在HE染色步骤中,机器在工作后期,随着染片工作量增大,脱水环节中乙醇浓度降低,对伊红着色有影响。

巢蛋白和α-抑制素蛋白在不同卵巢性索-间质肿瘤中的表达

目的:探讨巢蛋白(Nestin)和α-抑制素(α-inhibin)蛋白在不同卵巢性索-间质肿瘤(SCST)中的表达情况。方法:选取卵泡膜细胞瘤、卵巢纤维瘤、成年型粒层细胞瘤、卵巢微囊性间质瘤和卵巢低分化支持-莱迪(Sertoli-Leydig)细胞瘤组织标本,制备石蜡切片,行苏木精-伊红染色和α-inhibin、Nestin免疫组织化学染色。采用Spearman法分析Nestin和α-inhibin表达评分的相关性。结果:Nestin在SCST中的卵泡膜细胞瘤、卵巢纤维瘤、成年型粒层细胞瘤中呈阴性表达,在卵巢微囊性间质瘤和卵巢低分化Sertoli-Leydig细胞瘤组织中Leydig细胞呈阳性表达。α-inhibin在卵泡膜细胞瘤、卵巢纤维瘤和成年型粒层细胞瘤呈阳性表达,在卵巢微囊性间质瘤和卵巢低分化Sertoli-Leydig细胞瘤组织中呈阴性表达。Nestin与α-inhibin表达评分无相关性(均P<0.05)。结论:卵泡膜细胞瘤、卵巢纤维瘤、成人型颗粒细胞瘤组织中Nestin蛋白阴性表达,α-inhibin蛋白阳性表达。卵巢微囊性间质瘤、卵巢低分化Sertoli-Leydig细胞瘤组织中Nestin蛋白阳性表达,α-inhibin蛋白阴性表达,且Nestin与α-inhibin蛋白表达无明显关系。

Hepatic and renal effects of oral stingless bee honey in a streptozotocin-induced diabetic rat model

BACKGROUND Diabetes is known damage the liver and kidney,leading to hepatic dysfunction and kidney failure.Honey is believed to help in lowering the blood glucose levels of diabetic patients and reducing diabetic complications.However,the effect of stingless bee honey(SBH)administration in relieving liver and kidney damage in diabetes has not been well-studied.AIM To investigate the effect of SBH administration on the kidney and liver of streptozotocin-induced(STZ;55 mg/kg)diabetic Sprague Dawley rats.METHODS The rats were grouped as follows(n=6 per group):non-diabetic(ND),untreated diabetic(UNT),metformin-treated(MET),and SBH+metformin-treated(SBME)groups.After successful diabetic induction,ND and UNT rats were given normal saline,whereas the treatment groups received SBH(2.0 g/kg and/or metformin(250 mg/kg)for 12 d.Serum biochemical parameters and histological changes using hematoxylin and eosin(H&E)and periodic acid–Schiff(PAS)staining were evaluated.RESULTS On H&E and PAS staining,the ND group showed normal architecture and cellularity of Bowman’s capsule and tubules,whereas the UNT and MET groups had an increased glomerular cellularity and thickened basement membrane.The SBH-treated group showed a decrease in hydropic changes and mild cellularity of the glomerulus vs the ND group based on H&E staining,but the two were similar on PAS staining.Likewise,the SBME-treated group had an increase in cellularity of the glomerulus on H&E staining,but it was comparable to the SBH and ND groups on PAS staining.UNT diabetic rats had tubular hydropic tubules,which were smaller than other groups.Reduced fatty vacuole formation and dilated blood sinusoids in liver tissue were seen in the SBH group.Conversely,the UNT group had high glucose levels,which subsequently increased MDA levels,ultimately leading to liver damage.SBH treatment reduced this damage,as evidenced by having the lowest fasting glucose,serum alanine transaminase,aspartate transaminase,and alkaline phosphatase levels compared to other groups,although the levels of liver enzymes were not statistically significant.CONCLUSION The cellularity of the Bowman’s capsule,as well as histological alteration of kidney tubules,glomerular membranes,and liver tissues in diabetic rats after oral SBH resembled those of ND rats.Therefore,SBH exhibited a protective hepatorenal effect in a diabetic rat model.

苯酚在病理技术苏木精-伊红染色法中对病理切片质量的影响

目的探究苯酚在病理技术苏木精-伊红(HE)染色法中对病理切片质量的影响。方法选取2022年3月至2023年1月赣州市妇幼保健院纳入的80例实施病理诊断的患者作为研究对象,按照随机数字表法分为观察组与对照组,每组各40例。对照组病理切片采用常规HE染色,观察组病理切片在对照组的基础上联合苯酚染色。评估两组患者病理切片诊断准确率、病理切片质量。结果观察组病理切片诊断准确率为92.50%,高于对照组的病理切片诊断准确率(75.00%),差异有统计学意义(χ^(2)=4.501,P=0.034)。观察组病理切片质量优良率为100.00%,高于对照组病理切片质量优良率(85.00%),差异有统计学意义(χ^(2)=4.505,P=0.034)。结论苯酚应用于病理技术HE染色效果明显,可有效提高病理切片诊断准确率以及病理切片质量,为临床诊断和治疗相关疾病提供科学合理技术手段。

苏木精-伊红染色切片评估食管鳞状细胞癌中三级淋巴结构的预后意义

目的探索利用苏木素-伊红(hematoxylin-eosin,HE)染色切片评估三级淋巴结构(tertiary lymphoid structures,TLS)在食管鳞状细胞癌(esophageal squamous cell carcinoma,ESCC)中的潜在预后预测价值。方法选取2014—2015年河南省肿瘤医院医院收治的未经过术前放化疗的87例ESCC患者的手术标本,收集临床病理数据。采用HE染色法检测肿瘤区域内TLS的存在情况、密度、最大直径和分布情况。采用Kaplan-Meier生存分析和多因素Cox回归分析评估TLS特征对ESCC患者预后的影响。收集肿瘤基因图谱(Cancer Genome Atlas,TCGA)数据库中84例ESCC患者的临床病理数据对结果进行验证。结果在收集的87例ESCC患者中,58例经检测到在TLS,与TCGA数据库中的84例患者中44例存在TLS的情况一致。TLS的存在及其平均密度、直径和分布与ESCC患者的生存时间延长相关。但在校正病理特征之后,仅TLS的存在状态为ESCC患者中长期生存的独立预后因素(HR=0.432,P=0.004),与TCGA数据库中验证的结果一致(HR=0.359,P=0.028)。结论在ESCC中常规利用HE染色法评估TLS的存在是可行的,且可以作为有效的预后预测生物标志物。

Preparation of Taenia solium Stained Specimens and Morphological Observation

[ Objective ] In order to observe the morphological structure of Taenia solium and improve the dyeing method of Taenia solium. [ Method ] Taenia soli- um was stained by hematoxylin-eosin （HE） staining method and observed under microscope. Taenia solium specimens with clear structure and bright color were made. [ Results] Male reproductive system and excretory system of Taenia solium segment could be observed under the optical microscope, structures of uterus and eggs in Taenia solium gravid proglottid could also be observed. [ Conclusion] Aiming at providing a reference for the identification of Taenia solium and the diagnosis of the tapeworm disease.

黏蛋白乳化剂在1418例细胞蜡块制备技术中的应用研究

目的探讨黏蛋白乳化剂制备的1418例非妇科细胞蜡块的苏木素伊红(HE)染色、免疫组织化学(IHC)染色的效果和诊断价值。方法收集2018年7月至2022年1月该院送检的1418例细胞学标本,离心,弃上清液,加入甲醛-乙醇(AF)液5 mL,黏蛋白乳化剂250μL,混匀,离心,弃上清液,取出沉淀细胞块放入包埋盒,按照常规石蜡切片制备程序进行固定、脱水、透明、包埋、切片和HE染色,对细胞块病理诊断为恶性的285例标本,切2138张片进行IHC染色检测,观察黏蛋白乳化剂制备的细胞块及切片的外观大体情况;探讨细胞块切片HE染色和IHC染色的显微镜下情况和切片优良率;以患者术后病理诊断结果为金标准,综合评估黏蛋白乳化剂制备的细胞块的诊断效能。结果(1)黏蛋白乳化剂制备的细胞块制作成功率是100.00%,细胞块外观呈圆形果冻样胶体的结构、细胞聚集成团、硬度适中。细胞块切片外观完整、呈规则的圆形结构、疏松度适中。(2)细胞块可连续均匀切片,HE染色背景清晰,核浆着色鲜艳,红蓝分明,对比清晰,细胞均匀分布于圆形区域内,细胞形态呈现立体感,细胞块切片厚薄均一,IHC染色背景干净,结构和细胞均呈特异性染色。(3)细胞块的HE染色切片优良率为98.94%;IHC染色切片优良率为99.30%。(4)细胞块诊断的灵敏度、特异度、符合率和约登指数分别是96.94%、100.00%、99.37%、0.9694。结论黏蛋白乳化剂制备细胞块的技术操作便捷,成本低,富集率高,形态好,细胞块石蜡切片可长期保存,其HE染色和IHC染色优良率高,具有良好的诊断效能,应用前景广阔。

环保型组织样本制备液在不同脂肪细胞检测中的应用价值

背景:当前使用的传统试剂(甲醛溶液固定、乙醇脱水、二甲苯透明脱蜡)对组织前期处理制作的常规石蜡切片质量较低,已成为制约脂肪细胞性肿瘤病理诊断的瓶颈。而且,传统试剂容易对实验室环境造成污染,对实验室人员造成多种健康危害。目的:探讨环保型组织样本制备液在脂肪细胞肿瘤苏木精-伊红染色及非典型性脂肪瘤性肿瘤/高分化脂肪肉瘤MDM2基因检测中的应用价值。方法:选取2016年2月至2022年7月期间592例脂肪细胞肿瘤标本为研究对象,同一标本对半切开,按照所使用的标本前期处理试剂的不同,随机分为2组:传统组和环保组。传统组使用传统试剂甲醛溶液固定、乙醇脱水、二甲苯透明脱蜡制作常规石蜡切片592张;环保组使用环保型组织样本制备液制作切片592张。依据切片苏木精-伊红染色质量的不同等级,比较两组切片的苏木精-伊红染色优良率;进一步对其中病理确诊为非典型性脂肪瘤性肿瘤/高分化脂肪肉瘤的33例标本再次切片后,使用荧光原位杂交法检测MDM2基因,比较两组切片MDM2基因扩增率的差异。结果与结论:(1)相对于传统组,环保组的脂肪细胞肿瘤的组织切片更舒展、更完整,细胞无折叠,染色更清晰,红蓝对比度更佳,细胞结构更致密;(2)环保组的组织切片苏木精-伊红染色优良率高于传统组,差异有显著性意义(P=0.000);(3)相对于传统组,环保组的MDM2探针和CSP12探针仅在标本染色体特定的区域内结合,更具特异性;(4)环保组杂交成功细胞数高于传统组,差异有显著性意义(P=0.000);(5)传统组和环保组的MDM2基因信号数、MDM2/细胞值、CSP12信号数、CSP12/细胞值、MDM2/CSP12值之间的差异无显著性意义(P>0.05);(6)两组MDM2基因扩增率比较,差异无显著性意义(P=0.31);(7)结果表明,环保型组织样本制备液有利于提高脂肪细胞肿瘤的苏木精-伊红染色优良率,保证了非典型性脂肪瘤性肿瘤/高分化脂肪肉瘤MDM2基因的检测质量,有临床推广使用的价值。

3种组织固定液对大菱鲆肝脏切片质量影响和效果比较

为查明3种组织固定液对大菱鲆肝脏石蜡切片质量的影响,筛选效果最佳的固定液。本实验选用波恩氏(Bouin)、戴维森氏(Davidson)和4%多聚甲醛(Paraformaldehyde, PFA)3种组织固定液,对大菱鲆的肝脏进行固定,制作石蜡切片,通过苏木素-伊红(H-E)和免疫组织化学(Immunohistochemistry)染色,观察组织细胞形态学特征,利用统计学方法分析细胞完整度和免疫组化阳性率情况。结果发现,3种固定液对大菱鲆肝组织石蜡切片固定效果存在显著差异,Bouin固定液组织H-E染色细胞结构清晰、形态完整,Davidson固定液H-E染色组织溶解、细胞结构不清晰,但能分辨出核质界线,4%多聚甲醛组H-E染色鲜艳,但核质分界不清晰。完整肝细胞比例,4%PFA显著低于Bouin和Davidson固定液(P<0.05),且Bouin和Davidson固定液无显著差异(P>0.05)。肝组织增殖细胞核抗原(proliefrating cell nuclear antigen, PCNA)免疫组化染色阳性率3种固定液存在显著差异,Bouin固定液显著高于4%PFA和Davidson(P<0.05)。综上,使用Bouin固定液对肝脏固定,进行H-E染色或免疫组化染色能观察到更为完整的组织细胞结构和细胞增殖情况,效果最佳。

Protective effect of liraglutide on the myocardium of type 2 diabetic rats by inhibiting polyadenosine diphosphate-ribose polymerase-1

BACKGROUND In recent years,studies have found that the occurrence and development of diabetic cardiomyopathy(DCM)is closely related to an increase in polyadenosine diphosphate-ribose polymerase-1(PARP-1)activity.PARP-1 activation could be involved in the pathophysiological process of DCM by promoting oxidative stress,the inflammatory response,apoptosis and myocardial fibrosis.AIM To investigate the mechanism of liraglutide in improving myocardial injury in type 2 diabetic rats,further clarified the protective effect of liraglutide on the heart,and provided a new option for the treatment of DCM.METHODS Forty healthy male SD rats aged 6 wk were randomly divided into two groups,a normal control group(n=10)and a model group(n=30),which were fed an ordinary diet and a high-sugar and high-fat diet,respectively.After successful modeling,the rats in the model group were fed a high-glucose and high-fat diet for 4 wk and randomly divided into a model group and an intervention group(further divided into a high-dose group and a low-dose group).The rats were fed a high-glucose and high-fat diet for 8 wk and then started drug intervention.Blood samples were collected from the abdominal aorta to detect fasting blood glucose and lipid profiles.Intact heart tissue was dissected,and its weight was used to calculate the heart weight index.Haematoxylin and eosin staining was used to observe the pathological changes in the myocardium and the expression of PARP-1 in the heart by immunohistochemistry.RESULTS The body weight and heart weight index of rats in the model group were significantly increased compared with those in the normal control group,and those in the intervention group were decreased compared with those in the model group,with a more obvious decrease observed in the high-dose group(P<0.05).In the model group,myocardial fibers were disordered,and inflammatory cells and interstitial fibrosis were observed.The cardiomyopathy of rats in the intervention group was improved to different degrees,the myocardial fibers were arranged neatly,and the myocardial cells were clearly striated;the improvement was more obvious in the high-dose group.Compared with the normal control group,the expression of PARP-1 in myocardial tissue of the model group was increased,and the difference was statistically significant(P<0.05).After liraglutide intervention,compared with the model group,the expression of PARP-1 in myocardial tissue was decreased,and the reduction was more obvious in the high-dose group(P<0.05)but still higher than that in the normal control group.CONCLUSION Liraglutide may improve myocardial injury in type 2 diabetic rats by inhibiting the expression of myocardial PARP-1 in a dose-dependent manner.

胸腹水细胞块切片结合免疫组化在病理诊断中的应用

目的 分析病理诊断中应用胸腹水细胞块切片+免疫组化的诊断价值。方法 200份临床送检胸腹水标本,均进行细胞块切片结合免疫组化、苏木精-伊红染色法(HE)检测。分析标本病理结果与检测阳性率,比较腺癌与恶性间皮瘤细胞角质蛋白7(CK7)、癌胚抗原(CEA)、Wilm’s肿瘤基因-1(WT-1)、钙结合蛋白(CR)抗体表达阳性情况。结果 200份临床送检胸腹水标本中,有40份疑似肿瘤性胸腹腔积液标本,病理诊断均为肿瘤性胸腹腔积液。细胞块切片结合免疫组化检测阳性率100.00%(40/40)高于HE染色检测的67.50%(27/40),差异有统计学意义(P<0.05)。腺癌CK7抗体表达阳性率100.00%、CEA抗体表达阳性率95.46%高于恶性间皮瘤的0、0,CR抗体表达阳性率4.54%低于恶性间皮瘤的75.00%,差异有统计学意义(P<0.05)。腺癌与恶性间皮瘤WT-1抗体表达阳性率比较,差异无统计学意义(P>0.05)。结论 胸腹水细胞块切片结合免疫组化临床病理诊断肿瘤细胞的阳性率高,有助于鉴别原发疾病类型、早期治疗。