Histocytochemistry

组织细胞化学
  • 文章类型: Journal Article
    特发性肺纤维化(IPF)是一种来源不明的进行性肺部疾病,治疗选择有限,预后不良。来自利用间充质干细胞(MSCs)的临床前研究的令人鼓舞的发现表明,它们可以作为管理慢性肺部疾病的有希望的治疗替代方案。比如IPF。这项研究的目的是比较骨髓来源的MSCs(BM-MSCs)与泼尼松龙的效率,标准的抗炎药,在博来霉素(BLM)诱导的肺纤维化大鼠中。创建了四组:对照组,BLM组,泼尼松龙治疗组,和BM-MSC处理组。诱导肺纤维化,气管内施用5mg/kg的BLM。BLM显著增加促炎细胞因子和氧化应激标志物的血清水平。光和透射电子显微镜研究也揭示了受干扰的肺结构。α-平滑肌肌动蛋白免疫表达上调,转化生长因子β-1和Bax得到证实。有趣的是,所有发现在使用泼尼松龙和BM-MSCs治疗时显著回归。然而,用BM-MSCs治疗的结果优于泼尼松龙。总之,BM-MSC可能是管理肺纤维化的有希望的方法。
    Idiopathic pulmonary fibrosis (IPF) is a progressive lung disease of unknown origin with limited treatment options and poor prognosis. The encouraging findings from preclinical investigations utilizing mesenchymal stem cells (MSCs) indicated that they could serve as a promising therapeutic alternative for managing chronic lung conditions, such as IPF. The objective of this study was to compare the efficiency of bone marrow-derived MSCs (BM-MSCs) versus prednisolone, the standard anti-inflammatory medication, in rats with bleomycin (BLM)-induced lung fibrosis. Four groups were created: a control group, a BLM group, a prednisolone-treated group, and a BM-MSCs-treated group. To induce lung fibrosis, 5 mg/kg of BLM was administered intratracheally. BLM significantly increased serum levels of pro-inflammatory cytokines and oxidative stress markers. The disturbed lung structure was also revealed by light and transmission electron microscopic studies. Upregulation in the immune expression of alpha-smooth muscle actin, transforming growth factor beta-1, and Bax was demonstrated. Interestingly, all findings significantly regressed on treatment with prednisolone and BM-MSCs. However, treatment with BM-MSCs showed better results than with prednisolone. In conclusion, BM-MSCs could be a promising approach for managing lung fibrosis.
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  • 文章类型: Journal Article
    凝集素是对在各种组织中表达的糖缀合物具有特异性亲和力的碳水化合物结合蛋白。凝集素作为研究有很大的用处,组织化学,和哺乳动物系统中的诊断工具。研究了斑马鱼肝脏中12种常用植物凝集素的反应性。四种凝集素,番茄凝集素(TL),小麦胚芽凝集素,伴刀豆球蛋白A,和Jacalin对肝实质结构表现出强烈的反应性。重要的是,TL与幼虫和成人肝内胆管网络片段内的糖缀合物反应,从胆管到胆管。我们提供的证据表明,凝集素可以作为研究斑马鱼肝脏结构和功能特征的重要组织化学工具。
    Lectins are carbohydrate-binding proteins with specific affinity to glycoconjugates expressed in various tissues. Lectins are of substantial utility as research, histochemical, and diagnostic tools in mammalian systems. Reactivity of 12 commonly used plant-based lectins was studied in zebrafish liver. Four lectins, tomato lectin (TL), wheat germ agglutinin, concanavalin A, and Jacalin showed strong reactivity to hepatic parenchymal structures. Importantly, TL reacted to glycoconjugates within segments of the larval and adult intrahepatic biliary network, from canaliculi to bile ducts. We provide evidence that lectins can serve as important histochemical tools to investigate the structural and functional characteristics of the zebrafish liver.
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  • 文章类型: Journal Article
    脑组织的生物矿化发生在正常和病理条件下。硬脑膜生物矿化很普遍,发生在1-72%的病例中,取决于患者的年龄和研究方法。在脑膜中肿瘤生长的条件下,生物矿物沉积物的数量只会增加,达到100%的情况下,膜膜脑膜瘤。由于钙化经常出现在脑膜中,钙化脑膜瘤的鉴别诊断存在问题。共有30个具有生物矿化致密结构迹象的脑膜瘤样本,特征性的紧缩,作为对照(II组),检查了无任何生物矿化迹象的假单胞菌体(I组)和30例脑膜瘤样本。为了检测病理性生物矿化,使用宏观描述的方法研究脑膜瘤组织,组织学,组织化学,和免疫组织化学,扫描电子显微镜与显微分析,和透射电子显微镜。显著较高水平的caspase3和成骨细胞标志物的表达特征(在I组中,较低水平的OPG表达和较高水平的RANKL的存在,SPARC的表达没有波动)可能表明脑膜瘤中生物矿物沉积物的营养不良类型。
    Biomineralization of brain tissues occurs both in normal and pathological conditions. Dura mater biomineralization is widespread and occurs in 1-72% of cases, depending on the patient\'s age and research method. The amount of biomineral deposits under the conditions of tumor growth in the meninges only increases, reaching 100% in the case of psammomatous meningiomas. Since calcifications are often found in the meninges, the problem of differential diagnosis with calcified meningiomas arises. A total of 30 samples of meningiomas with signs of biomineralization-dense structure, characteristic crunch, psammoma bodies (group I) and 30 samples of meningiomas without any signs of biomineralization were examined as controls (group II). To detect pathological biomineralization, the meningioma tissue was studied using the methods of macroscopic description, histology, histochemistry, and immunohistochemistry, scanning electron microscopy with microanalysis, and transmission electron microscopy. A significantly higher level of caspase3 and features of the expression of osteoblastic markers (a lower level of OPG expression and a higher level of the presence of RANKL in group I, the absence of fluctuations in the expression of SPARC) may indicate a dystrophic type of development of biomineral deposits in meningiomas.
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  • 文章类型: Journal Article
    (1)研究背景:矢车菊是一种药用植物,其花卉在中草药中应用广泛。该研究的目的是定位负责花瓣中分泌产物产生的花组织,并分析挥发性化合物。迄今为止,尚未对该物种花的挥发性化合物进行研究。(2)方法:光,荧光,扫描和透射电子显微镜技术用于研究。使用组织化学测定将亲脂性化合物定位于组织中。使用固相微萃取(SPME)和气相色谱-质谱(GC-MS)测定挥发性化合物。(3)结果:研究表明花瓣实质中分泌的产生,其超微结构具有分泌组织的特征。亲脂性分泌物位于薄壁组织的细胞和细胞间隙以及表皮细胞的壁和表面,它通过角质层微通道释放后积累。发现倍半萜是挥发性化合物的主要成分,其中β-石竹烯含量最高(26.17%),α-腐殖烯含量最高(9.77%)。(4)结论:鉴于树脂中经常出现一些挥发性成分(石竹烯,δ-cadinene)和表皮表面丰富的分泌残留物,我们假设花释放的花青C.分泌物是树脂质混合物(含油树脂),这常见于植物中,如文献数据所示。这种分泌可能在花青草花的治疗作用中起重要作用。
    (1) Background: Centaurea cyanus L. is a medicinal plant whose flowers are widely used in herbal medicine. The aim of the study was to localise flower tissues that are responsible for the production of secretory products in petals and to analyse the volatile compounds. The volatile compounds of the flowers of this species have not been investigated to date. (2) Methods: Light, fluorescence, scanning and transmission electron microscopy techniques were used in the study. Lipophilic compounds were localised in the tissues using histochemical assays. Volatile compounds were determined with the use of solid phase microextraction (SPME) and gas chromatography-mass spectrometry (GC-MS). (3) Results: The study showed production of secretion in the petal parenchyma, whose ultrastructure has features of a secretory tissue. The lipophilic secretion was localised in the cells and intercellular spaces of the parenchyma and in the walls and surface of epidermal cells, where it accumulated after release through cuticle microchannels. Sesquiterpenes were found to constitute the main group of volatile compounds, with the highest content of β-caryophyllene (26.17%) and α-humulene (9.77%). (4) Conclusions: Given the presence of some volatile components that are often found in resins (caryophyllene, delta-cadinene) and the abundant secretion residues on the epidermal surface, we suppose that the C. cyanus secretion released by the flowers is a resinaceous mixture (oleoresin), which is frequently found in plants, as shown by literature data. This secretion may play an important role in the therapeutic effects of C. cyanus flowers.
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  • 文章类型: Journal Article
    这项研究是为了调查大体解剖结构,组织化学,高加索松鼠胃的免疫组织化学。总共包括五只雄性松鼠。获得的结果表明,胃的形状是急性新月形,由四个区域组成,即心脏,眼底,身体,和幽门.胃腺及其壁由四层组成;浆膜,肌层,粘膜下层,和粘膜。粘膜的所有部分都被简单的柱状上皮覆盖。此外,胃腺为简单管状。眼底和身体区域的粘液细胞为PAS阳性。在胃腺的下三分之一,顶叶,并确定了主要细胞。主要细胞的数量朝向幽门区增加。扫描电子显微镜显示,眼底区域包含许多皱褶和凹槽,每个皱褶都带有许多腺体开口。在身体区域,细胞呈多边形并形成网状结构。在幽门区,发现了身体和胃窦之间的粘膜脊和较小曲率上的厚粘膜峰。免疫组织化学染色显示Sox9在眼底区域强表达,然而,其在体内和幽门中的表达为中度至微弱。总之,高加索松鼠的胃与包括啮齿动物在内的其他哺乳动物有一些异同。获得的结果可能会增强我们对该物种的摄食习惯和消化系统组织卫生学的基本知识。
    This study was performed to investigate the gross anatomy, histochemistry, and immunohistochemistry of the stomach in the Caucasian squirrel. A total of five male squirrels were included. The obtained results showed that the shape of the stomach was acutely crescent and consisted of four regions namely; cardiac, fundus, body, and pylorus. The stomach glandular and its wall was consisted of four layers; serosa, muscularis, submucosa, and mucosa. All part of the mucosa was covered by a simple columnar epithelium. Furthermore, gastric glands were of the simple tubular type. The mucous cells of the fundic and body regions were PAS-positive. In the lower third of the gastric glands, parietal, and chief cells were identified. The number of chief cells was increased toward the pyloric area. Scanning electron microscopy showed that the fundus region contained many plicae and grooves and each plica carried numerous glands opening. In the body region, cells were polygonal and formed network-like structures. In the pyloric region, a mucosal ridge between the body and antrum and a thick mucosal crest on the lesser curvature were identified. Immunohistochemical staining showed that Sox9 was strongly expressed in the fundic region, however, its expression in the body and pylorus was moderate to weak. In conclusion, there are some similarities and differences between the stomach of the Caucasian squirrel and other mammals including rodents. The obtained results may enhance our basic knowledge regarding the feeding habits and digestive system histophysiology of this species.
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  • 文章类型: Journal Article
    背景:通常对从细针抽吸(FNA)标本制备的细胞块进行辅助研究。对唾液腺(SG)FNA的细胞块的辅助研究应用研究有限。这项多机构研究评估了对细胞块进行的辅助研究在SG病变诊断中的作用。以及它们对临床管理的影响。
    方法:在三个大型学术机构的电子病理学档案中搜索SGFNA,并对细胞块进行辅助研究。病人的人口统计,FNA网站,细胞学诊断,辅助研究,并记录手术随访情况。如果需要,根据米兰唾液腺细胞病理学报告系统(MSRSGC)对细胞学诊断进行了重新分类.
    结果:确定了117例SGFNA病例,包括MSRSGCI类的3、10、11、6、23、4和60例,II,III,IVa,IVb,V,VI,分别进行手术随访,每个类别范围为27%至100%。辅助研究,包括组织化学,免疫细胞化学(IHC),原位杂交(ISH)在每个类别中60%-100%的病例中都是有益的。在可疑的恶性肿瘤(V)和恶性(VI)类别中,恶性肿瘤的风险均为100%。辅助研究改善了60%非肿瘤性病例的诊断(II,6/10),100%的良性肿瘤病例(IVa,6/6),和98.3%的恶性肿瘤(VI,59/60)。
    结论:对具有足够材料的SGFNA细胞块进行明智和基于病例的辅助研究,可以通过进一步表征非典型/肿瘤细胞来提高诊断率。特别是在MSRSGC类别IVa-VI。
    BACKGROUND: Ancillary studies are commonly performed on cell blocks prepared from fine-needle aspiration (FNA) specimens. There are limited studies in application of ancillary studies on cell blocks from salivary gland (SG) FNAs. This multi-institutional study evaluates the role of ancillary studies performed on cell blocks in the diagnosis of SG lesions, and their impact on clinical management.
    METHODS: The electronic pathology archives of three large academic institutions were searched for SG FNAs with ancillary studies performed on cell blocks. The patient demographics, FNA site, cytologic diagnosis, ancillary studies, and surgical follow-up were recorded. If needed, the cytologic diagnoses were reclassified as per the Milan System for Reporting Salivary Gland Cytopathology (MSRSGC).
    RESULTS: 117 SG FNA cases were identified including 3, 10, 11, 6, 23, 4, and 60 cases in MSRSGC categories I, II, III, IVa, IVb, V, VI, respectively with surgical follow-up available ranging from 27% to 100% within each category. Ancillary studies including histochemistry, immunocytochemistry (IHC), and in situ hybridization (ISH) were beneficial in 60%-100% of cases in each category. Risk of malignancy was 100% in both the suspicious for malignancy (V) and malignant (VI) categories. Ancillary studies improved diagnosis in 60% of non-neoplastic cases (II, 6/10), 100% of benign neoplasm cases (IVa, 6/6), and 98.3% of malignant cases (VI, 59/60).
    CONCLUSIONS: Judicious and case-based ancillary studies performed on SG FNA cell blocks with sufficient material can improve the diagnostic yield by further characterization of the atypical/neoplastic cells, particularly in MSRSGC categories IVa-VI.
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  • 文章类型: Journal Article
    背景:早期食管鳞状细胞癌(ESCC)的细胞学检测仍然具有挑战性。因此,我们引入了一种快速细胞学筛查方法,并评估了其作为早期ESCC和前体病变的内镜检查前筛查的疗效.方法:该方法由海绵样本检索组成,自动液基细胞学治疗和载玻片制备,计算机辅助筛查和手动诊断。评估了早期ESCC和前体病变的检测功效。此外,将诊断效率与人工诊断进行比较.结果:83例早期ESCC和前体病变患者和2,090例无症状的ESCC高风险参与者被纳入。整个过程在两个工作日内完成。83例患者均检测到异常细胞,在2,090名无症状参与者中,有272名(13.01%)受试者。早期ESCC,高级别上皮内瘤变,低度上皮内瘤变和反流性食管炎和正常的内镜检查结果在8、13、11、187和53名异常细胞参与者中检测到,分别。计算的灵敏度,特异性,检测早期ESCC和前兆病变的阳性预测值和阴性预测值为100%,88.34%,11.76%,100%,分别。与手动诊断相比,该方法在较短的持续时间内完成(5.4±0.45分钟vs320.2±132.4分钟,p<0.001),更高的诊断准确性(96.7%vs74.4%,p=0.015)和更好的观察者间协议(93.3%vs66.7%,K=0.286,p<0.001)。结论:我们的研究为早期ESCC和前体病变的内镜检查前筛查提供了一种有前途的方法。
    Background: Cytological detection of early esophageal squamous cell carcinoma (ESCC) remains challenging. Therefore, we introduced a rapid cytological screening method and evaluated its efficacy as a pre-endoscopy screening for early ESCC and precursor lesions. Methods: This method consisted of a sponge sample retrieval, automatic liquid-based cytological treatment and slides preparation, computer-assisted screening and manual diagnosis. Efficacy for detection of early ESCC and precursor lesions was evaluated. Also, diagnostic efficiency was compared with manual diagnosis. Results: Eighty-three patients with early ESCC and precursor lesions and 2,090 asymptomatic participants with high risks of ESCC were enrolled. Whole procedure was accomplished within two working days. Abnormal cells were detected in all 83 patients, and in 272 (13.01%) subjects among 2,090 asymptomatic participants. Early ESCC, high-grade intraepithelial neoplasia, low-grade intraepithelial neoplasia and reflux esophagitis and normal endoscopic findings were detected in 8, 13, 11, 187 and 53 participants with abnormal cells, respectively. The calculated sensitivity, specificity, positive predictive value and negative predictive value for detection of early ESCC and precursor lesions were 100%, 88.34%, 11.76%, and 100%, respectively. Compared with manual diagnosis, this method was accomplished in a shorter time duration (5.4 ± 0.45 min vs 320.2 ± 132.4 min, p < 0.001), a higher diagnostic accuracy (96.7% vs74.4%, p = 0.015) and a better inter-observer agreement (93.3% vs66.7%, K = 0.286, p < 0.001). Conclusions: Our study provides a promising methodology as pre-endoscopy screening for early ESCC and precursor lesions.
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  • 文章类型: Journal Article
    通过酶组织化学在所有神经节的低温恒温器切片上研究了猪腰脊神经节(LSG)结构中三肽基肽酶I(TPPI)活性的分布,使用底物甘氨酰-L-氨酰-L-甲硫基-5-氯-1-蒽醌酰肼(GPM-CAH)和4-硝基苯甲醛(NBA)作为可视化因子。光学显微镜观察显示几乎所有LSG结构中的TPPI活性。半定量比较了不同细胞类型中的酶反应。在小神经元中观察到强烈的反应,卫星神经节细胞和一些神经纤维。在大型感觉体神经元中发现了弱反应性,而对TPPI的中度反应是在中部感觉体神经元和一些神经纤维中确定的。单因素方差分析的统计学分析显示,每平方毫米的TPPI阳性神经元数量没有显著差异(当p为0.05时)。通过酶组织化学方法获得的原始数据使我们有理由假设TPPI积极参与猪LSG中所有神经元结构的功能。根据我们的结果,可以认为TPPI活性对于自主神经和躯体感觉神经元的功能很重要。
    Distribution of tripeptidyl peptidase I (TPPI) activity in the structures of porcine lumbar spinal ganglia (LSG) was studied by enzyme histochemistry on cryostat sections from all the ganglia using the substrate glycyl-L-prolyl-L-methionyl-5-chloro-1-anthraquinonyl hydrazide (GPM-CAH) and 4-nitrobenzaldehyde (NBA) as visualization factor. Light microscopic observations showed TPPI activity in almost all the LSG structures. The enzyme reaction in different cell types was compared semi-quantitatively. Strong reaction was observed in the small neurons, satellite ganglia cells and some nerve fibers. Weak reactivity was found in the large sensory somatic neurons, whereas moderate reaction for TPPI was determined in the middle sensory somatic neurons and some nerve fibers. Statistical analysis by one-way ANOVA showed no significance of difference (when p⟨0.05) for the number of TPPI positive neurons per mm2. The original data obtained by the enzyme histochemistry method give us a reason to presume that TPPI actively participates in the functions of all the neuronal structures in porcine LSG. According to our results, it could be suggested that TPPI activity is important for the functions of autonomic and somatic sensory neurons.
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  • 文章类型: Journal Article
    This study describes the morphology and histochemistry of the digestive tube of the herbivorous fish Rhytiodus microlepis, which is commonly found in the Amazonian floodplain lakes, Brazil. Thirty-eight fish were used in this study. The digestive tube was measured and dissected for anatomical description, and stained with (i) haematoxylin and eosin, for histological analyses; (ii) periodic acid-Schiff, for detected neutral mucins; and (iii) Alcian Blue (AB) pH 1.0 and 2.5 for acid and sulphated acid mucins. The oesophagus, with its secretory cells and taste buds, is adapted for lubrication of vegetable matter, resistance to friction and reception of stimuli. The stomach is U-shaped and rich in gastric glands, particularly in cardiac and fundic regions, which are adapted to receive large amounts of food, and promotes the digestion. The intestine comprises more than 70% of the digestive tube. All portions of the intestine, except the rectum, have the same pattern of mucosal folds, and the goblet cells reacted positively to all histochemical methods (PAS, AB 1.0 and 2.5), while rodlet cells reacted only to periodic acid-Schiff (PAS) staining. Neutral mucins and sulphated acid mucins predominated in the anterior portion of the intestine and a high concentration of carboxylated acid mucosubstances were present in the other portions. The rectum showed a strong reaction to all histochemical methods. The muscular layer of the intestine consists of three layers, which showed features presumably related to the rigid nature of the food. A simple squamous epithelium constitutes the serous layer over the entire length of the tube.
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  • 文章类型: Journal Article
    不可逆电穿孔(IRE)是目前用于治疗实体瘤的局部非热消融技术。这里,我们研究了上消化道内窥镜下IRE的临床效力和安全性。猪在食道中使用最近设计的内窥镜IRE导管进行电穿孔,胃,和十二指肠。引入了两个连续的策略来优化消化道的电能。首先,每个器官进行电穿孔和能量放大,以确认上限能量诱导不适当的组织结果,包括出血和穿孔。从第一步中排除不可接受的能量,在逐步减少能量的情况下进行连续电穿孔,以确定损伤每一层的能量.对不适当的电强度的前瞻性研究导致每个组织超过一定能量阈值的广泛出血和肠穿孔。然而,在排除的能量以下进行的实验伴随苏木精和伊红染色和末端脱氧核苷酸转移酶dUTP缺口末端标记分析显示,受损的粘膜面积和深度随着能量的减少而显著减少.相关的组织病理学检查显示,电穿孔病变处有炎性细胞浸润,并有固缩核。这项研究证明了内镜IRE在粘膜发育不良或中空内脏早期恶性肿瘤中的可能性。
    Irreversible electroporation (IRE) is a local non-thermal ablative technique currently used to treat solid tumors. Here, we investigated the clinical potency and safety of IRE with an endoscope in the upper gastrointestinal tract. Pigs were electroporated with recently designed endoscopic IRE catheters in the esophagus, stomach, and duodenum. Two successive strategies were introduced to optimize the electrical energy for the digestive tract. First, each organ was electroporated and the energy upscaled to confirm the upper limit energy inducing improper tissue results, including bleeding and perforation. Excluding the unacceptable energy from the first step, consecutive electroporations were performed with stepwise reductions in energy to identify the energy that damaged each layer. Inceptive research into inappropriate electrical intensity contributed to extensive hemorrhage and bowel perforation for each tissue above a certain energy threshold. However, experiments performed below the precluded energy accompanying hematoxylin and eosin staining and terminal deoxynucleotidyl transferase dUTP nick-end labeling assays showed that damaged mucosal area and depth significantly decreased with decreased energy. Relevant histopathology showed infiltration of inflammatory cells with pyknotic nuclei at the electroporated lesion. This investigation demonstrated the possibility of endoscopic IRE in mucosal dysplasia or early malignant tumors of the hollow viscus.
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