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@#Objective To explore the key points and difficulties of intraoperative frozen section diagnosis of pulmonary diseases. Methods The intraoperative frozen section and postoperative paraffin section results of pulmonary nodule patients in Beijing Chaoyang Hospital, Capital Medical University from January 2021 to January 2022 were collected. The main causes of misdiagnosis in frozen section diagnosis were analyzed, and the main points of diagnosis and differential diagnosis were summarized. Results According to the inclusion criteria, a total of 1 263 frozen section diagnosis results of 1 178 patients were included in the study, including 475 males and 703 females, with an average age of 58.7 (23-86) years. In 1 263 frozen section diagnosis results, the correct diagnosis rate was 95.65%, and the misdiagnosis rate was 4.35%. There were 55 misdiagnoses, including 18 (3.44%) invasive adenocarcinoma, 17 (5.82%) adenocarcinoma in situ, 7 (35.00%) mucinous adenocarcinoma, 4 (2.09%) minimally invasive adenocarcinoma, 3 (100.00%) IgG4 related diseases, 2 (66.67%) mucinous adenocarcinoma in situ, 1 (16.67%) atypical adenomatous hyperplasia, 1 (14.29%) sclerosing pulmonary cell tumor, 1 (33.33%) bronchiolar adenoma, and 1 (100.00%) papillary adenoma. Conclusion Intraoperative frozen section diagnosis still has its limitations. Clinicians need to make a comprehensive judgment based on imaging examination and clinical experience.
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OBJECTIVES@#The advanced non-small cell lung cancer (NSCLC) patients with pleural effusion have no opportunity for surgery treatment. Epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs) are the first-line drugs for these patients with EGFR-sensitive mutation. However, the disease progression and drug update during or after treatment of EGFR-TKIs bring more challenges and puzzles to clinical diagnosis and treatment, which inevitably requires archived pleural cell samples for EGFR re-examination or comparative study. Understanding the DNA quality of archived pleural fluid samples and effectively using archival data of pleural fluid cells are of great significance for tracing the origin of cases and basic medical research. This study aims to evaluate the consistency of EGFR mutant gene expression between the 2 methods, and to explore a reliable way for preserving cytological data and making full use of cytological archival data via cell HE staining smear and cell paraffin section.@*METHODS@#A total of 57 pleural fluid cytology cases in the Department of Pathology of China Aerospace Center Hospital from October 2014 to April 2021 were selected. Tumor cells were detected by cell HE staining smears and immunohistochemical staining for TTF-1 and Napsin A in the paired cell paraffin sections. There were more than 200 tumor cells in cell HE staining smear and the proportion of tumor cells were ≥70% in matched cell paraffin sections. Patients with 2 cell smears (one for cell data retention and the other for DNA extraction) were selected as the research subjects, and 57 pleural fluid samples were enrolled. EGFR gene mutation was detected by amplification refractory mutation system-polymerase chain reaction in 57 paired cell HE staining smears and cell paraffin sections. DNA concentration was 2 ng/μL. Cell HE smear was amplified side-by-side with DNA samples from paired cell paraffin sections. Result determination was according to the requirements of the reagent instructions. The external control cycle threshold (Ct) value of the No. 8 well of the samples to be tested was between 13 and 21, which was considered as successful and reliable samples. When the Ct value of EGFR gene mutation was <26, it was considered as positive; when the Ct value was between 26 and 29, it was critical positive; when the Ct value was equal or more than 29, it was negative. ΔCt value was the difference between mutant Ct value and externally controlled Ct value. The smaller the ΔCt value was, the better the quality of DNA of the detected sample was.@*RESULTS@#Among the 57 pleural effusion samples, 42 patients were hospitalized with pleural effusion as the first symptom, accounting for 73.7% (42/57). EGFR mutation was detected in 37 samples [64.9% (37/57)]. The mutation rate for 19del was 37.8% (14/37) while for L858R was 48.6% (18/37). Females were 56.7% (21/37) of mutation cases. The mutation consistency rate of cell HE staining smear and matched cell paraffin sections was 100%. The ΔCt values of cell HE staining smears were less than those of matched cell paraffin sections. The mutation Ct values of 37 cytological samples were statistically analyzed according to the preservation periods of the years of 2014-2015, 2016-2017, 2018-2019, and 2020-2021. There were significant differences in cell paraffin section in the years of 2014-2015 and 2016-2017 compared with the years of 2018-2019 and 2020-2021, while no significant differences were found in cell HE staining smear. Statistical analysis of externally controlled Ct values of 57 cytological samples showed that there were significant differences between cell HE staining smears and cell paraffin section in the years of 2014-2015 and 2016-2017, compared with the years of 2018-2019 and 2020-2021. The mutational Ct values of 37 paired cell blocks and smears were all <26, and the externally controlled Ct values of 57 paired cell paraffin sections and HE staining smears were all between 13 and 21.@*CONCLUSIONS@#The DNA quality of cell HE smears and matched cell paraffin section met the qualified requirements. Two methods possess show an excellent consistency in detecting EGFR mutation in NSCLC pleural fluid samples. The DNA quality of cell HE staining smear is better than that of cell paraffin sections, so cell HE staining smear can be used as important supplement of the gene test source. It should be noted that the limitation of cell HE staining smears is non-reproducibility, so multiple smears of pleural fluid are recommended to be prepared for multiple tests.
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Female , Humans , Male , Carcinoma, Non-Small-Cell Lung/drug therapy , DNA Mutational Analysis/methods , ErbB Receptors/genetics , Lung Neoplasms/drug therapy , Mutation , Paraffin/therapeutic use , Pleural Effusion/genetics , Protein Kinase Inhibitors/therapeutic use , Staining and LabelingABSTRACT
Objective:To explore the advantages and disadvantages of frozen section versus rapid paraffin section in the evaluations of donor organ.Methods:Five cases of donor liver and 8 cases of discarded donor kidney were collected from 2017 to 2021.Tissues were harvested and prepared by frozen section, rapid paraffin section and normal paraffin section.After hematoxylin-eosin (H&E) staining, the specimens of donor kidney/liver were evaluated by differential histopathological structures and donor quality scoring system.Results:Rapid paraffin section was similar to normal paraffin section in reflecting the proportion of glomerulosclerosis (18.6%±22.3%), arteriolar hyaline degeneration (43.7%±23.8%) and arteriolar stenosis (47.9%±29%). The proportion of glomerulosclerosis (0.8%±2.2%), arteriolar hyaline degeneration (4.9%±7.4%) and arteriolar stenosis (5.3%±7.5%) were lower in frozen sections than those in rapid paraffin sections.The diagnoses of hydropic degeneration and necrosis in donor liver were more accurate in rapid paraffin section.Conclusions:Rapid paraffin section is superior to frozen section in observing histopathological changes under microscope.Scoring of donor organ is more precise according to rapid paraffin section.
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Objective To solve the problem that the rat retina paraffin sections are easily exfoliated from the slides and each layer are easily separation and fracture,we need to find a way to improve the retina paraffin section method and evaluate the tissue fixation. Methods We used 4 fixation liquids including 10%paraformaldehyde,4%paraformaldehyde, 4% paraformaldehyde and 95% alcohol and glacial acetic acid mixed liquid (FAA fixatiue solution ) combined with paraformaldehyde to fix the retinal tissue, and observed the fixation efficacy under microscope after HE staining. Results The effects of 10%paraformaldehyde and 4%paraformaldehyde fixed samples showed moderate separation and fracture of retina,but the HE staining retinal slices pre-treated by the FAA fixafive solution had bright and uniform color,although occasionally some parts of the retina were exfoliated from the slide, but it was not easy to take off,and had complete structure without separation and rupture. Conclusion The retina paraffin section fixed by FAA ixafive solution with 4% paraformaldehyde is superior to pure paraformaldehyde, and the paraformaldehyde concentration has no obviously influence on HE staining results.
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ObjectiveTo develop a method of location of prenatal female mouse reproductive tract based on paraffin block serial sections of female fetuses, and get high quality paraffin sections of fetuses containing female reproductive tract (FRT).And to investigate the role of Wnt5a during the development of FRT of prenatal mice. Methods The sex of fetuses at gravidity 15.5 days( G15.5d), G17.5d, G19.5d) and G21.5d was identified by polymerase chain reaction(PCR),and the female fetuses were embedded in paraffin block, the specimen was sectioned serially . One of every four sections was stained by hematoxylin-eosin(HE), and the next one being used later.The images of specimen were taken with digital camera and the serial sections were obtained. The paramesonephric duct /uterus were located and recognized. Immunohistochemistry was used to determine the location and intensity of Wnt5a staining in the middle of the paramesonephric duct / uterus. Results The morphology of the paramesonephric duct /uterus was recognized first and the sections of fetuses with paramesonephric ducts were obtained. The intensity of Wnt5a immunohistochemical staining was increased from G15.5d to G21.5d in mesenchymal cells(P<0.01).Conclusion Wnt5a plays a marked role in early period of female mouse reproductive tract, and is possible to be a key factor to induce uterus differentiation and development.
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Thirty five breast cancer cases had paraffin section biopsies while 22 had a frozen followed immediately by surgery. Those who underwent biopsy at surgery showed a better survival and freedom from the disease. A time delay of not more than one week apparently seems not to have been detrimental to most of our patients. However, the ideal is a frozen followed by amputation, even only for economical and psychological reasons, for it spares the patient anxious waiting days and reduces her hospital expenses. Minimizing infection by an umbrella of antibiotics after a paraffin section biopsy and gentle handling of the growth can help prevent acceleration of the lymph flow, thereby prevent early regional metastases.(Author)
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Objective With personal computer(PC) as the platform,to explore an computerized 3D reconstruction method based on rat's cervical cord serial sections. Methods Normal cervical cord segment of SD rat was embeded in paraffin block,four locating marks were made at four corners around the specimen,the specimen was sectioned serially with Leica paraffin microtome,after each three sections,the images of specimen and outer locating marks were taken with CANON digital camera and the serial sections were obtained.The obtained images were ordered,cut,turned into gray scale images and background uneven corrected.The edges of white matter,grey matter and locating marks in each image were extracted,the locating marks were used for automated registration of serial sections with 3D-DOCTOR software,the 3D surface reconstruction of cervical cord segment was observed,cut and measured freely in PC. Results The grey matter,white matter of rat cervical cord segment were reconstructed from serial transverse sections,the reconstructed segment was measured to obtain the surface area and volume data.Conclusion The paraffin embedding and "hard" locating marks techniques can be used to obtain the images of serial transverse sections of rat's cerival cord segment,to make 3D reconstruction which can be observed and measured freely.