Quantification in histopathology—Can magnetic particles help?

Every year, more than 270,000 people are diagnosed with cancer in the UK alone; this means that one in three people worldwide contract cancer within their lifetime. Histopathology is the principle method for confirming cancer and directing treatment. In this paper, a novel application of magnetic particles is proposed to help address the problem of subjectivity in histopathology. Preliminary results indicate that magnetic nanoparticles cannot only be used to assist diagnosis through improving quantification but also potentially increase throughput, hence offering a way of dramatically reducing costs within the routine histopathology laboratory.     

Measurement of nitric oxide production by the lesioned rat retina with a sensitive nitric oxide electrode

Nitric oxide (NO) plays multiple roles in the nervous system. It is produced as a result of damage or injury of the retina as a part of the central nervous system. Detailed knowledge of the extent and the time course of NO production is of great importance for the understanding of pathological processes and their appropriate medical treatment.Sections of rat retina were stained with antibodies against the three isoforms of NO synthase (NOS) at several time points after a lesion of the optic nerve. No significant changes of NOS expression could be seen at any of the checked time points.For the electrochemical detection of NO production, we modified small platinum electrodes with a NO-sensitive nickel porphyrin by electrochemical polymerisation. Compared to other substances, electrochemically polymerised eugenol was found to be most suitable for protection against interferences. For the measurements, differential pulse amperometry was used. The response to nitric oxide was linear.NO production of adult rat retinas was measured post axotomy after different time points with electrochemical electrodes ex vivo. With non-treated retinas, an NO concentration of approximately 15 M was measured. NO concentration is elevated after an axotomy reaching its highest value of up to 30 M 5 days after the lesion. The NO concentration is decreased below the initial value after 9–14 days post axotomy.     

Towards the Expression of Sex Hormone Receptors in the Human Vocal Fold

BACKGROUND: The human larynx is assumed to be a steroid receptor target organ. There are only very limited data on the evidence of steroid receptors in the vocal folds, although voice alterations due to hormonal influence and treatment have been found. GOAL OF THE STUDY: To investigate the expression of estrogen alpha, progesterone, and androgen receptors in human vocal folds (vocalis muscle, glands, lamina propria, epithelium). METHODS: Immunohistochemically, vocal fold cadaver specimens of 15 autopsied patients (6 women, 9 men), which were taken approximately 4 to 8 hours postmortem were investigated. Furthermore, one (male) vocal fold biopsy obtained intraoperatively during a laryngectomy was tested. RESULTS: No specific immunohistochemical staining for the different types of steroid hormones investigated could be observed in either the postmortem taken biopsies nor the intraoperatively one. However, several unspecific staining patterns could be observed. CONCLUSION: The results of this study contradict recently published data and question the expression of sex hormone receptors in the vocal folds. Main causes of false interpretations of unspecific staining are discussed.     

Ultrastructural localization of NADPH diaphorase and nitric oxide synthase in the neuropils of the snail CNS

Comparative studies on the nervous system revealed that nitric oxide (NO) retains its function through the evolution. In vertebrates NO can act in different ways: it is released solely or as a co-transmitter, released from presynaptic or postsynaptic site, spreads as a volumetric signal or targets synaptic proteins. In invertebrates, however, the possible sites of NO release have not yet been identified. Therefore, in the present study, the subcellular distribution of the NO synthase (NOS) was examined in the central nervous system (CNS) of two gastropod species, the terrestrial snail, Helix pomatia and the pond snail, Lymnaea stagnalis, which are model species in comparative neurobiology. For the visualization of NOS NADPH-diaphorase histochemistry and an immunohistochemical procedure using a universal anti-NOS antibody were applied. At light microscopic level both techniques labeled identical structures in sensory tracts ramifying in the neuropils of central ganglia and cell bodies of the Lymnaea and Helix CNS. At ultrastructural level NADPH-d reactive/NOS-immunoreactive materials were localized on the nuclear envelope and membrane segments of the rough and smooth endoplasmic reticulum, as well as the cell membrane and axolemma of positive perikarya. NADPH-d reactive and NOS-immunoreactive varicosities connected to neighboring neurons with both unspecialized and specialized synaptic contacts. In the varicosities, the majority of the NADPH-d reactive/NOS-immunoreactive membrane segments were detected in round and pleomorph agranular vesicles of small size (50–200 nm). However, only a small portion (16%) of the vesicles displayed the NADPH-d reactivity/NOS-immunoreactivity. No evidence for the postsynaptic location of NOS was found. Our results suggest that the localization of NADPH-diaphorase and NOS is identical in the snail nervous system. In contrast to vertebrates, however, NO seems to act exclusively in an anterograde way possibly released from membrane segments of the presynaptic transmitter vesicle surface. Based on the subcellular distribution of NOS, NO could be both a volume and a synaptic mediator, in addition NO may function as a co-transmitter.     

5- 脂氧合酶在小鼠恶唑酮溃疡性结肠炎中的表达研究

目的 研究5- 脂氧合酶（5-LOX）在恶唑酮诱导溃疡性结肠炎（UC）小鼠结肠黏膜中的表达情况,探讨其在UC 发病 机制中的作用及其与炎症程度的关系。方法 50只Balb/c小鼠分为正常对照组10 只和模型组40只。模型组用3%恶唑酮致敏2d,第5 天用0.5%恶唑酮灌肠。第8 天用颈椎脱臼法处死所有小鼠。实验期间观察小鼠疾病活动指数（DAI）,处死小鼠后行结肠组织学病理评分（HPS）。RT-PCR 法检测结肠黏膜5-LOXmRNA 的表达,免疫组织化学方法检测结肠黏膜5-LOX 的蛋白表达情况。结果 模型组DAI 和HPS 均高于对照组,5-LOXmRNA和蛋白表达也明显高于对照组（均P＜0.01）,并随炎症程度加重,表达水平明显增高。结论 5-LOX 在UC 中表达水平明显升高,并参与了UC 的发生和发展,在一定程度上可反应疾病的活动度,并且可能作为药物治疗的潜在靶点。     

多梳基因Bmi-1 在甲状腺乳头状癌中的表达及意义

目的 研究多梳基因Bmi-1 在甲状腺乳头状癌组织中的表达及临床意义。方法 分别采用免疫组化法、Westernblot 法和RT-PCR 法检测30 例甲状腺乳头状癌组织、30 例结节性甲状腺肿组织及30 例正常甲状腺组织中多梳基因Bmi-1 及其mRNA 的表达情况,并分析Bmi-1与甲状腺乳头状癌临床病理特征的关系。结果 免疫组化法检测发现Bmi-1 在甲状腺乳头状癌组织中阳性表达率73.3%,与结节性甲状腺肿（36.7%）及与正常组织（16.7%）的阳性表达率相比差异有统计学意义（P＜0.05）。Western blot 法检测甲状腺乳头状癌中Bmi-1 表达量与结节性甲状腺肿组织及正常组织相比,差异有统计学意义（P＜0.05）。RT-PCR法检测甲状腺乳头状癌中Bmi-1mRNA 表达量与结节性甲状腺肿组织及正常组织相比,差异有统计学意义（P＜0.05）。Bmi-1 阳性表达在甲状腺乳头状癌有无淋巴结转移中的差异有统计学意义（P＜0.05）,在年龄、性别、肿瘤大小、分期等差异无统计学意义。结论 Bmi-1 在甲状腺乳头状癌中表达升高,可能在甲状腺乳头状癌的发生、发展过程中起重要作用,并可望作为甲状腺乳头状癌临床诊断的标记物之一。     

c-Met蛋白在甲状腺癌中的表达及其临床病理意义

目的 检测c-Met 蛋白在甲状腺癌及癌旁甲状腺组织中的表达,并分析c-Met 蛋白表达与甲状腺乳头状癌患者临床病理特征的关系。方法 采用免疫组织化学En Vision二步法检测81 例甲状腺癌组织及52 例癌旁甲状腺组织中c-Met 蛋白的表达。结果 甲状腺癌和癌旁甲状腺组织中c-Met 蛋白阳性率分别为46.9%（38/81）和7.7%（4/52）,两者比较有统计学差异（P＜0.01）。其中甲状腺乳头状癌中c-Met 蛋白阳性率50.8%（31/61）,髓样癌中阳性率60.0%（3/5）,均高于滤泡癌26.7%（4/15）,但均无统计学差异（均P＞0.05）。甲状腺乳头状癌c-Met蛋白表达与患者各项临床病理特征均无相关性（均P＞0.05）。结论 甲状腺癌中c-Met 表达增强,其高表达可能与甲状腺癌发生相关,有望成为抗甲状腺癌治疗的一个潜在靶点。     

Effect of gambogenic acid in attenuating diethylnitrosamine (DEN)-induced hepatocellular carcinoma in rat model

Liver cancer, specifically hepatocellular carcinoma has been a widespread problem among general population. This study aims to investigate the modulating mechanism of gambogenic acid, a phenolic xanthonoid, in diethylnitrosamine (DEN)-induced liver cancer in rats. Male Wistar albino rats were clustered into four groups (n = 6). Group I served as control treated with normal saline. Hepatocellular carcinogenesis was induced in rats by single intraperitoneal (i.p.) administration of DEN in saline (200 mg/kg b.w.) for groups II and III. Group III received oral administration of gambogenic acid (20 mg/kg b.w.) one hour post DEN administration, whereas group IV received oral administration of gambogenic acid (20 mg/kg b.w.) alone. Rats were sacrificed after 16 weeks to determine the levels of hepatic biomarkers, oxidative stress markers, hematological profile and histopathological changes. Gambogenic acid significantly ameliorated the expressions of oxidative stress markers TBARS, GSH (P < 0.05), enzymatic antioxidants GPx, CAT, SOD, GST (P < 0.05), apoptosis mediators (P < 0.05), and serum biomarkers for liver damage and tumor formation (P < 0.05) compared with DEN-induced model group. Hepatocellular levels of 8-OHdG were significantly diminished (P < 0.05) by gambogenic acid against the damage incurred by DEN. Liver histopathological derangements caused by DEN were reversed by gambogenic acid. The results clearly impacted the effect of gambogenic acid in attenuating DEN-induced hepatocellular carcinoma in rats mediated through NF-kβ pathway and hepatocellular oxidative damage.     

Effects of Micronutrients on Oxidative Stress and Islet Function in Type 1 Diabetes Mellitus Rats： Relationships to Th2 Type Cytokines, Interleukin-4, and Interluekin-10

To observe the effects of the micronutrients on oxidative and autoimmune destruction of islets so as to prevent a person from the onset and development of type 1 Diabetes Mellitus(T1DM),the interleukin-4 and interleukin-10 expressions of lymphocytes in peripheral blood and spleen of T1DM rats were determined by flow cytometry.GSH-Px activity and MDA level in the rats' pancreas were measured using biochemical methods.The insulin contents in serum and β cell insulin secret storage were tested by RIA and IHC,respectively.There was an increase in the percentages of IL-4 and IL-10 positive lymphocytes in the peripheral blood and spleen of the groups of rats supplemented with various combinations of micronutrients(p<0.01 and p<0.05,respectively);the blood glucose concentration decreased(p<0.05);both the functional β cell in islets and the insulin content in pancreatic tissue increased(p<0.05 and p<0.01);the GSH-Px activity and MDA level of pancreas in the rats enhanced and decreased respectively(p<0.01 and p<0.05).The results suggest that micronutrients may alleviate the islet lesions by upregulating the expressions of IL-4 and IL-10 and lowering oxidative stress in diabetic rats.