The value of circulating tumor cells with positive centromere probe 8 in the diagnosis of small pulmonary nodules.

Background: Circulating cancer cells (CTCs) provide opportunities for early diagnosis and evaluation of cancer stage as a more acceptable non-invasive liquid biopsy. The advanced development and use of CTCs for diagnosis or prognosis just started in recent years.Methods: Fifty three patients, diagnosed as SPNs with a diameter less than 30 mm by CT examination, was enrolled for statistical analysis based on their CTCs level, CT examination features, serum tumor marker concentrations, and histopathological characteristics. Centromere probe 8 (CEP8) was utilized as a marker for CTCs determination. Results: The CTCs level was significantly different between malignant and benign SPNs, as well as between early (0/Ⅰa) and advanced (Ⅰb/Ⅱ/Ⅲ) stages of lung cancer. ROC analysis showed that the CTCs level had diagnostic effect on malignant SNPs. Combined use of CTCs and density feature of CT morphology further improve the overall diagnostic effect on the pTNM stage of these SPNs determined as lung cancer (≤Ia vs. >Ia stage) compared to use of these two markers solely, especially increased the diagnostic specificity. Moreover, in bigger, single, and solid SPNs based on CT morphology, the CTCs level significantly correlated with the malignant histopathology. Additionally, triple staining (CEP8, EpCAM and CKs) results using samples from 22 out of 53 patients showed that more CTCs was detected when CEP8 was used as a marker. Conclusions: The CTCs determined by CEP8 positive would be a potential adjuvant diagnostic marker for the malignance and stage of lung cancer for patients with SPNs.

The value of circulating tumor cells with positive centromere probe 8 in the diagnosis of small pulmonary nodules.

Background Circulating cancer cells (CTCs) provide opportunities for early diagnosis and evaluation of cancer stage as a more acceptable non-invasive liquid biopsy. The advanced development and use of CTCs for diagnosis or prognosis just started in recent years. Methods Fifty three patients, diagnosed as SPNs with a diameter less than 30 mm by CT examination, was enrolled for statistical analysis based on their CTCs level, CT examination features, serum tumor marker concentrations, and histopathological characteristics. Centromere probe 8 (CEP8) was utilized as a marker for CTCs determination. Results The CTCs level was significantly different between malignant and benign SPNs, as well as between early (0/Ia) and advanced (Ib/II/III) stages of lung cancer. For the malignancy judgment and the pTNM stage diagnosis of SPNs, ROC analysis results showed that combined use of CTCs level or density feature of CT morphology significant improved diagnostic effect compared to use of these two markers solely. Moreover, in bigger, single, and solid SPNs based on CT morphology, the CTCs level significantly correlated with the malignant histopathology. Additionally, triple staining (CEP8, EpCAM and CKs) results using samples from 22 out of 53 patients showed that more CTCs was detected when CEP8 was used as a marker. Conclusion The CTCs determined by CEP8 positive would be a potential adjuvant diagnostic marker for the malignance and stage of lung cancer for patients with SPNs.

Mucosubstances in the porcine gastrointestinal tract: Fixation, staining and quantification

Mucins are of great interest in intestinal research and histochemical methods are often employed to identify them. Since it is in the nature of mucins that they are “hard to hold onto” once they come into contact with water, a frequently used medium in histochemistry, there are a number of challenges that may decrease diagnostic accuracy. As the outcome of methods published for microscopic detection of mucosubstances proved to be unsatisfactory in our hands, the aim was the establishment of a reliable and reproducible protocol. Tissue samples were available from pig feeding experiments. In the present study, we focus on a fixation / staining procedure without making comparisons between differently fed pigs. Several fixation and staining procedures were evaluated for their use in semiautomatic quantification and quality assessment of different mucus fractions simultaneous on one tissue section. Cryostat sectioning, subsequent fixation steps with heat, ethanol and modified Bouin’s solution, followed by triple staining with high iron diamine, alcian blue and periodic acid-Schiff turned out to be the best method to identify sulfomucin, sialomucin and neutral mucin simultaneous on one tissue section. This methodology resulted in very good morphology of goblet cells with intact mucin containing vesicles within the cells, which was comparable to ultrastructural electron microscopical observations. Semiautomatic quantification of different mucins was possible. In conclusion, reliable mucus quantification and assessment of mucus quality requires strictly tested procedures. According to our experience, the most important aim after cryosectioning is fast fixation of the mucosubstances, which requires a combination of different fixation steps.

Immunohýstochemýcal staýnýng of cd3, cd79ácy and s-100 on bursa fabrýcýus, thymus and spleen of turkeys (Meleagrýs Gallapavo)

Distribution of CD3, CD79ácy and S-100, immunohistochemically, in the bursa of Fabricius, thymus and spleen has not been reported in turkeys. Therefore, we determined the localisation of anti-CD3 protein antibodies for mature T-lymphocytes, anti-CD79ácy antibodies for B-lymphocytes and anti-S-100 protein antibodies for follicular dentritic cells in the bursa of Fabricius, thymus and spleen, which are the lymphoid organs, in a study sample of turkeys. Triple staining method was applied to demonstrate the general structure. Moreover, in all the organs positive reactions were observed with the CD3, CD79ácy and S-100 antibodies. It was also found that similar areas had a positive reaction with CD79ácy and S-100 in all of the tested organs. It was remarkable that CD79ácy reacted positively on Hassall’s corpuscles (strongly) and the reticular cells (weakly) in the medulla of the thymus instead of the B-lymphocyte positive areas.