OBJECTIVE: To investigate paraffin section thickness in immunohistochemical detection of p16 expression in cervical tissue samples. METHODS: p16 immunohistochemical staining was performed in 150 cases of chronic cervicitis, 126 cases of low-grade squamous intraepithelial lesions(LSIL), 96 cases of high-grade squamous intraepithelial lesions (HSIL) and 78 cases of cervical cancer from January 2014 to March 2018 in Zhongshan Boai Hospital. The results of p16 protein expression in paraffin sections with thickness of 2, 3, 4, 5 and 6 μm were compared using Logistic regression analysis. RESULTS: With the increase of slice thickness, the staining of p16 protein in nucleus gradually increased. The thickness of cervical slices in chronic cervicitis and cervical cancer samples had no significant effect on the positive rate of p16 protein(=7.817 and 1.332, both >0.05), while the thickness of slices in LSIL and HSIL samples had significant effect on the positive rate of p16 protein (=17.688 and 10.182, <0.05 or <0.01). The stable and reliable results were obtained when the slices were between 3 and 5 μm thick. CONCLUSIONS Paraffin sections with thickness of 3.0-5.0 μm are recommended for immnohistochemical staining of p16 protein in cervical tissue samples.
Biomarkers, Tumor ; genetics ; metabolism ; Cervical Intraepithelial Neoplasia ; Cyclin-Dependent Kinase Inhibitor p16 ; genetics ; metabolism ; Female ; Histocytological Preparation Techniques ; standards ; Humans ; Immunohistochemistry ; Paraffin ; Squamous Intraepithelial Lesions of the Cervix ; Uterine Cervical Neoplasms ; physiopathology

Ascitic Fluid ; cytology ; Cytological Techniques ; instrumentation ; methods ; Equipment Design ; Histocytological Preparation Techniques ; instrumentation ; methods ; Humans ; Paraffin Embedding ; Specimen Handling ; instrumentation ; methods

OBJECTIVE: To analyze and discuss the feasibility of rabbit carotid artery treated with decellularization and photo-oxidation. METHODS: Sixty vascular slices of rabbit carotid artery were divided into a fresh group, a cryopreservation group, a glutaraldehyde group, and a decellularization plus photo-oxidation group 15 in each group. To evaluate the physical properties of all the rabbit carotid arteries by testing heat-shrinking temperature, tensile stress and the max elongation of each group. Then by buliding subcutaneous embedding model in SD rats we evaluated the biological stability and the anti-calcification function property of the above rabbit carotid arteries, and the detection means included HE stain, atomic absorption spectrometry and Von-Kossa calcium salt stain. RESULTS: The heat-shrinking temperature, tensile stress and the max elongation in the cryopreservation group were lower or shorter than those of the other groups and the difference had statistical significance (P<0.05). Although the heat-shrinking temperature and the tensile stress in the decellularization plus photo-oxidation group were lower or shorter than those in the glutaraldehyde group (P<0.05), the max elongation in the decellularization plus photo-oxidation group was much longer than that in the glutaraldehyde group (P<0.05). The rabbit carotid artery treated with decellularization plus photo-oxidation showed lower immunogenicity and better biological stability and better anti-calcification property compared with the other groups. CONCLUSION Decellularization associated with photo-oxidation is a suitable and novel protocol for small caliber artery allograft with a diameter of less than 6 mm which is unbreakable to mechanical properties and conducive to biological stability, which has a broad prospect.
Animals ; Blood Vessel Prosthesis ; Calcinosis ; prevention & control ; Carotid Arteries ; cytology ; transplantation ; Cell Separation ; methods ; Female ; Histocytological Preparation Techniques ; Male ; Oxidants, Photochemical ; pharmacology ; Oxidation-Reduction ; Rabbits ; Rats ; Rats, Sprague-Dawley ; Transplantation, Heterologous

This study was aimed to establish a smear protocol for preparing bone marrow cells and investigate its effect on fluorescence in situ hybridization (FISH) signal. Probe DNA (C-myc, MDM2, STK6) was labeled with Spectrum Green, PromoFluor-555 and PromoFluor-415 by nick translation. Five bone marrow samples were tested by two methods separately. Traditional method: after removing the erythrocytes by hypoosmotic solution, the bone marrow cells were fixed in methanol/acetic acid (3:1). Improved method: erythrocytes were removed using density gradient centrifugation and fixed in methanol. The samples were then fixed again in 2 formaldehyde for 5 min. The FISH signal was assessed by comparing the relative signal intensity of each fluorophore with the autofluorescence background. The results indicated that improved method greatly increased the ratio of fluorescence signal intensity in the Spectrum Green, PromoFluor-555 and PromoFluor-415 channel (traditional method: 4.3 ± 0.19, 3.52 ± 0.04, 3.07 ± 0.08; improved method: 9.89 ± 0.41, 7.55 ± 0.5, 5.67 ± 0.18, n = 5, P < 0.01) respectively. The signal intensity increased 2.32, 2.14 and 1.85-fold in the Spectrum Green, PromoFluor-555 and PromoFluor-415 channel respectively. In addition, the improved method decreased the split signals [traditional method: (15.8 ± 1.74), (20.42 ± 2.88), (23.2 ± 3.02); improved method: (8.6 ± 1.2), (12.28 ± 1.33), (12.6 ± 2.56), n = 5, P < 0.05]. It is concluded that the improved optimal procedure which facilitates FISH intensity on bone marrow cells is developed, showing potential for wide application in the diagnosis of hematologic diseases.
Bone Marrow Cells ; cytology ; Histocytological Preparation Techniques ; Humans ; In Situ Hybridization, Fluorescence ; methods

Hepatocytes ; pathology ; Histocytological Preparation Techniques ; methods ; Humans ; Liver ; pathology ; Liver Cirrhosis ; pathology

Objective. To study the clinical spectrum of Filipino patients with Williams Syndrome and to confirm the gene deletion by FISH analysis. Methods. From June 2005 to September 2008, patients who were seen at the Genetics clinic of the UP-PGH and who met the clinical criteria for Williams Syndrome were analyzed for the 7q11.23 deletion through karyotyping and FISH studies. A detailed history and a thorough dysmorphologic examination were performed. Relevant investigations included two-dimensional echocardiography, renal ultrasonography, ophthalmologic examination, developmental assessment and serum calcium determination. Result. Eight patients were included in the study. The mean age at first diagnosis was 8.5 years. All cases were sporadic. The chromosomal analysis was normal for all patients and in the FISH analysis, a 7q11.23 deletion was detected in 100% of cases. Distinctive facial features, cardiac abnormalities and developmental delay were present in all patients. The typical behavior of overfriendliness was observed in the majority of cases. Hypercalcemia was documented in only one case and no renal anomalies were detected. Conclusion. The craniofacial features were similar among patients but there is a broad spectrum of severity of clinical features in cardiovascular abnormalities, personality, behavior traits and mental capacity.
CYTOGENETICS ; GENETICS ; WILLIAMS SYNDROME ; NERVOUS SYSTEM DISEASES ; NEUROLOGIC MANIFESTATIONS ; NEUROBEHAVIORAL MANIFESTATIONS ; INTELLECTUAL DISABILITY ; GENE DELETION ; IN SITU HYBRIDIZATION, FLUORESCENCE ; AORTIC STENOSIS, SUPRAVALVULAR ; DIAGNOSIS ; DIAGNOSTIC TECHNIQUES AND PROCEDURES ; CLINICAL LABORATORY TECHNIQUES ; CYTOLOGICAL TECHNIQUES ; HISTOCYTOLOGICAL PREPARATION TECHNIQUES ; STAINING AND LABELING ; IN SITU HYBRIDIZATION ; ;

OBJECTIVEThe purpose of this study was to explore the value of thin-layer cytology (TLC) in intraoperative fine needle aspiration cytology diagnosis of pancreatic cancer.

METHODSResults of cytological examination with thin-layer smears were compared with that with conventional smears in intraoperative fine-needle aspiration (FNA) biopsy.

RESULTSTotally 271 fine needle aspiration biopsies were performed, among them, 70 were examined with thin-layer smears, showing unsatisfactory smear in 5 cases (7.1%); 201 were examined with conventional smears (CS), showing unsatisfactory smear in 9 cases (4.5%). No significant difference in the unsatisfactory smears was observed between those two groups. The positive rate of diagnosis with CS smears was 60.0% (42/70) and that of TLC was 81.6% (164/201), with a significant difference (P < 0.01). The sensitivity of CS and TLC was 68.9% and 87.7%, respectively (P < 0.01). The sensitivity of both FNA and frozen section diagnosis in 20 cases was 90.0%, respectively, but that of FNA combined with frozen section diagnosis was 95.0%. 9 cancer cases diagnosed by pathology were initially negative by cytology, but adenocarcinoma cells were found in 7 cases of them by the second time cytology examination.

CONCLUSIONThe positive rate is high in TLC smears, and unsatisfactory rate is low. TLC smears are one of the best methods in intraoperative confirmation of pancreatic cancer. The use of FNA smears combined with frozen section biopsy can further improve the sensitivity of diagnosis.

Adult ; Aged ; Aged, 80 and over ; Biopsy, Fine-Needle ; methods ; Cytodiagnosis ; Female ; Frozen Sections ; Histocytological Preparation Techniques ; methods ; Humans ; Intraoperative Period ; Male ; Middle Aged ; Pancreatic Neoplasms ; diagnosis ; pathology ; Young Adult

OBJECTIVETo investigate the reliability, practicability and limitations of cytopathological scrape method in the diagnosis of superficial lesions of skin and mucosal membrane.

METHODSThe quantity of cells harvested by different collecting methods were compared in 18 cases of malignant tumors of the skin and mucosal membrane. The scrape method was performed in 456 cases, of which 148 cases had corresponding histological specimens.

RESULTSGlass slide scraping provides the most satisfactory cell numbers from the lesions of skin and mucosal membrane. The specificity for diagnosing benign lesion in 37 patients was 100% and that for diagnosing malignant tumors in 111 patients was 92.8% (103). The overall diagnostic accuracy was 94.6% (140/148). The false negative rate, false positive rate and the overall misdiagnostic rates were 7.2% (8/111) , 0, and 5.4% (8/148) respectively.

CONCLUSIONSFor cytopathological examination of superficial skin and mucosal membrane lesions, the glass slide scraping methodprovides broad applications with high diagnostic accuracy and advantages of simplicity, safety and efficiency. Certain limitations do exist, but can be resolved by histological examination of the lesion.

Biopsy, Needle ; Breast Neoplasms ; pathology ; Carcinoma, Basal Cell ; pathology ; Carcinoma, Squamous Cell ; pathology ; Cytodiagnosis ; Female ; Histocytological Preparation Techniques ; methods ; Humans ; Lip Neoplasms ; pathology ; Male ; Mouth Mucosa ; pathology ; Mouth Neoplasms ; pathology ; Neoplasms ; pathology ; Paget's Disease, Mammary ; pathology ; Skin Neoplasms ; pathology ; Tongue Neoplasms ; pathology

Histocytological Preparation Techniques ; Humans ; Immunohistochemistry ; Microscopy, Electron ; methods ; Molecular Biology ; methods ; Staining and Labeling

The task of segmenting histiocyte is a crucial step in the analysis of histiocytic images which is an important application of computer vision to histopathology. The algorithm presented in this article was composed of two steps: (1) the morph-based preprocessing; (2) the ameliorated watershed method. In the first step, the difference between histiocytes was magnified in order to increase the visibility from the view of the computer vision, and then the ameliorated terrace-flooding-simulated watershed method was used to achieve the segmentation of histiocytic images in the second step. To test the performance of the algorithm, different samples of visual quality were tested and the result figures proved successful.
Algorithms ; Bone Marrow Cells ; cytology ; Cartilage ; cytology ; Histocytological Preparation Techniques ; methods ; Humans ; Image Processing, Computer-Assisted ; methods ; Numerical Analysis, Computer-Assisted ; Pattern Recognition, Automated ; methods