Clone 8g7g3 1

A 30-year (1980-2013) retrospective review of the cytology slides from our cytology division of pathology department revealed a total of 750 PTCs. Out of these, 73 cases of CPTC, which were confirmed by histologic sections, were retrieved from the files: 55 females and 18 males, with an age range of 28-52 years. There was no pediatric case. The cyst sizes ranged between 2.8 cm and 3 cm. Aspiration was performed from the cysts tissue (42 cases under ultrasound guidance and the remaining without guidance). The cysts yielded 2-3 mL of the fluid that was hemorrhagic in 40 cases and clear yellow in color in the remaining cases. The smears were prepared from the sediment of cytocentrifuged specimens. The air-dried smears were stained with the Wright–Giemsa stain, and alcohol-fixed smears were stained with Papanicolaou stain. Some selected cases were subjected to immunocytochemistry for thyroid transcription factor-1 (TTF-1, Dako, clone 8G7G3/1, Denmark), cytokeratin 19 (CK 19) (Dako, Clone RCK108, Denmark), and cluster of differentiation 68 (CD68) (Dako, clone KP1, Denmark).
The FNA smears of other cystic thyroid lesions were retrieved from the files for the comparative study. These cases included 300 colloid goiters, 290 adenomatoid nodules, 11 follicular neoplasms, and 9 hurtle cell neoplasms.
Medical ethics committee of our university approved the study.

For immunohistochemical characterization staining of whole tissue slides with antibodies directed against Pan-Cytokeratin (DAKO, Clone MNF116, Hamburg, Germany), CD56 (Zytomed, clone RCD56, Berlin, Germany), Chromogranin A (Novocastra, clone 5H7, Wetzlar, Germany) and TTF-1 (DAKO, Clone 8G7G3/1, Hamburg, Germany) was performed. The proliferation index was determined by staining with antibodies directed against the proliferation associated marker ki67 (Zytomed, clone K-2, Berlin, Germany) using a fully automated stainer (Dako AutostainerPlus). Apoptosis was determined by immunohistochemistry against cleaved caspase 3 (Cell Signalling, Beverly, MA, USA). The immunoexpression of ki67 was quantified by applying a four stage score (0= 0%, 1= <10%, 2=11 -50%, 3=> 50%), whereas cleaved caspase was scored by a different system (0 = 0%, 1= <5%, 2= 5-10%, 3= > 10%). Because PSMB4 was differentially expressed in any pulmonary NET subtype, the protein expression was assessed by immunohistochemistry (Sigma, polyclonal antibody, Taufkirchen, Germany).

First, 4-µm slices were dyed with hematoxylin and eosin (HE), and the histological type was analyzed by selecting different markers using IHC. After dewaxing in xylene and rehydrating in gradient ethanol, the slides were prepared for antigen retrieval by boiling them in 10-mM sodium citrate buffer. Then, 3% hydrogen peroxide was used to block the endogenous peroxidase activity of the slides. Thereafter, the samples were incubated with the following primary antibodies at 4 °C overnight using thyroid transcription factor 1 (TTF-1) (clone 8G7G3/1, DAKO; dilution 1:200), napsin A (polyclone, Abcam; dilution 1:250), P40 (polyclone, Abcam; dilution 1:200), cluster of differentiation 56 (CD56, clone 123C3, DAKO; dilution 1:100), synaptophysin (Syn, clone DAK-SYNAP, DAKO; dilution 1:50), cytokeratin 5/6 (CK5/6, clone D5/16 B4, DAKO; dilution 1:100), and chromogranin A (CgA, clone DAK-A3; dilution 1:200). The slides were subsequently incubated with an anti-primary antibody and then stained with diaminobenzidine and hematoxylin. Finally, they were dehydrated in ethanol, washed in xylene, and then covered with coverslips for further microscopic observation.