Immunocytochemical Examination Research Articles

Expression of osteopontin and osteocalcin in Osteoblast cells exposed to a combination of polymethylmethacrylate (PMMA) and hydroxyapatite (HAp): A prospective observational study.

The success of implant placement will depend on the ability of the implant material to integrate with the surrounding tissue. Polymethylmethacrylate (PMMA) has been used as an implant material, but it has several fallback properties in its interaction with bone tissue. The addition of hydroxyapatite (HAp) to PMMA is expected to produce reinforced bioceramic polymers with better mechanical and biological properties. The purpose of this study was to evaluate the expression of osteopontin and osteocalcin in cultured osteoblasts when exposed to two implant candidate materials: PMMA-HApGMP, derived from bovine bone and processed under Good Manufacturing Practice by a Tissue Bank, and PMMA-HApBBK, sourced from limestone (CaCO3) and processed by Balai Besar Keramik. Twenty-four fetal rat calvariae osteoblast cell cultures were randomly divided into 6 groups: 7- and 14-day control group, 7 and 14 days PMMA-HApGMP group, 7 and 14 days PMMA-HApBBK group. The expression of osteopontin and osteocalcin was seen by immunocytochemical examination. The results showed that the average expression of osteopontin and osteocalcin in the treatment group on the 7th and 14th days was higher than the control group. The expression of osteopontin and osteocalcin in the PMMA-HApGMP group increased significantly (P < .05) on day 14. The PMMA-HAp combination material can accelerate the process of osteoblast differentiation which is characterized by an increase in osteopontin and osteocalcin which are markers of bone formation. This will support in increasing osseointegration.

Interleukin-8/CXCR1 Signaling Contributes to the Progression of Pulmonary Adenocarcinoma Resulting in Malignant Pleural Effusion.

Pulmonary adenocarcinoma (PADC) treatment limited efficacy in preventing tumor progression, often resulting in malignant pleural effusion (MPE). MPE is filled with various mediators, especially interleukin-8 (IL-8). However, the role of IL-8 and its signaling mechanism within the fluid microenvironment (FME) implicated in tumor progression warrants further investigation. Primary cultured cells from samples of patients with MPE from PADC, along with a commonly utilized lung cancer cell line, were employed to examine the role of IL-8 and its receptor, CXCR1, through comparative analysis. Our study primarily assessed migration and invasion capabilities, epithelial-mesenchymal transition (EMT), and cancer stem cell (CSC) properties. Additionally, IL-8 levels in MPE fluid versus serum, along with immunohistochemical expression of IL-8/CXCR1 signaling in tumor tissue and cell blocks were analyzed. IL-8/CXCR1 overexpression enhanced EMT and CSC properties. Furthermore, the immunocytochemical examination of 17 cell blocks from patients with PADC and MPE corroborated the significant correlation between upregulated IL-8 and CXCR1 expression and the co-expression of IL-8 and CXCR1 in MPE with distant metastasis. In summary, the IL-8/ CXCR1 axis in FME is pivotal to tumor promotion via paracrine and autocrine signaling. Our study provides a therapeutic avenue for improving the prognosis of PADC patients with MPE.

Fine needle aspiration biopsy of parathyroid; is it meaningful? A cytologic study of 81 cases with histological and clinical correlations.

Recognizing the parathyroid gland and distinguishing the parathyroid from thyroid lesions in fine needle aspiration (FNA) is challenging. This study aimed to identify cytomorphologic features suggestive of parathyroid origin and to assess the utility of cytopathology in conjunction with ancillary tests in the identification of parathyroid glands. Ultrasound (US) guided FNA of parathyroid gland and lesions in 81 patients were reviewed concerning clinical history and correlated to histopathologic findings in available cases. FNA smears were evaluated for cellularity, architectural patterns, cellular and nuclear features, and background of the smears. In 78 cases, FNA was supplemented by a measurement of parathormone (PTH) levels in the needle washout fluid (FNA-PTH assay) and/or GATA3/PTH/chromogranin-A immunostainings. Sixty-four cases were diagnosed cytologically as parathyroid lesions in conjunction with FNA-PTH assay and/or immunocytochemical examinations. In an additional nine cases, a diagnosis of parathyroid lesions was rendered after repeated FNA with FNA-PTH assay. The histolopathologic diagnosis of surgically excised cases (n = 75) included parathyroid adenoma (60 cases), atypical parathyroid adenoma (4 cases), parathyroid hyperplasia (10 cases), and parathyroid carcinoma (1 case). Major cytological findings of parathyroid tissue included high cellularity, scattered naked nuclei, cribriform and three-dimensional clusters, stippled chromatin, and oxyphilic cytoplasm while papillary pattern or colloid-like material was identified in three cases respectively. No nuclear grooves or inclusions were seen in any case. High cellularity scattered naked nuclei, cribriform and three-dimensional patterns, stippled chromatin and oxyphilic cytoplasm are cytomorphologic features that favour parathyroid origin. A combination of these features with FNA-PTH assay and/or GATA3, PTH, and chromogranin-A immunostainings on cytologic specimens aid in the identification of parathyroid glands and the distinguishing of parathyroid from thyroid lesions.

Primary Hepatic Burkitt Lymphoma

The liver is an uncommon primary site for lymphoma, comprising only 0.4% of extranodal lymphomas, and liver involvement is almost always secondary to systemic lymphoma. Primary hepatic lymphoma has been reported to occur more frequently in immunosuppressed patients, especially with chronic hepatitis C infection, and may be associated with hepatitis B infection and the Epstein-Barr virus. Patients usually present with pain or a mass in the right upper quadrant, with or without jaundice, and it is more common in middle-aged males. The diagnosis of extranodal lymphoma or primary hepatic lymphoma can be difficult, especially in people living with human immunodeficiency virus (HIV) positivity or acquired immunodeficiency syndrome (AIDS). Due to its rarity, it is often overlooked as a possible differential diagnosis, with nonspecific imaging findings and a definitive diagnosis made only after anatomopathological and immunocytochemical examination. We present a rare case of Burkitt's lymphoma presenting as liver masses.

İnsan Trofoblast Hücre Hattında Sialik Asit Bağlayıcı Reseptörlerin (Siglecs) Ekspresyonu

Purpose: Cell surface proteins known as Siglecs exhibit a specific affinity for sialic acid. Primarily located on the surface of immune cells, Siglecs belong to the subset of lectins called I-type lectins. Siglecs have important roles in maternal-fetal immune tolerance. We aimed to analyze the expression of Siglecs on Human Villous Trophoblasts (HVT) cells.&#x0D; Materials and methods: Total RNA was extracted from the HVT cell line, cDNA was synthesized, and real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR) was performed the determine the changes in Siglec -3, -5, -6, -7, -8, -9, -10, -11, and -16 mRNA levels. In addition, Siglec levels were assessed by using immunohistochemical staining. Immunoreactivity against Siglec-6 and Siglec-9 was evaluated separately according to the intensity of brown color. &#x0D; Results: Expression levels of Siglec genes by qRT-PCR and melting curve analyses were performed using RNA extracted from the HVT cell line. Siglec -3, -5, -6, -7, -9, -10, -11, and -16 genes were found to be expressed in the HVT cell line. Differently, Siglec-8 results were undetected after cycle 40, which was considered a negative result. The immunocytochemical examination of the HVT cell line revealed that Siglec-6 expression was moderate in the cytoplasm (Score:2). Siglec-9 expression was prominent in the cytoplasm (Score:3). &#x0D; Conclusion: The results showed that Siglec-6 and Siglec-9 were expressed more than other Siglec proteins in the human trophoblast cells. Immunocytochemistry results also support these findings. Our study is the first to show Siglec-9 expression in human trophoblast cells.

Determination of Autophagy in Human Cervicovaginal Smears by Cytological and İmmunocytochemical Methods.

Autophagy is a catabolic process whereby organelles and long-lived proteins are recycled through lysosomes to maintain cellular homeostasis. This process is being widely studied using culture techniques and animal models; however, cervicovaginal smears have not been used to detect autophagy. Our study aims to detect and evaluate autophagy in normal, malignant, infectious, and atypical cells in cervicovaginal smears by using cytological and immunocytochemical methods. Papanicolaou-stained 200 cervicovaginal smears were examined and 55 of 200 (27.5%) smears containing negative for intraepithelial lesion or malignancy (NILM) with identifiable infections and/or reactive/reparative changes (INF); briefly, NILM-INF (n = 31, 56.4%), atypical (n = 4, 7.3%), and malignant cells (n = 20, 36.3%) were evaluated as a study group. One hundred forty-five of 200 (72.5%) normal smears were accepted as the NILM without any identifiable infections (control group). The autophagy marker protein Microtubule-associated protein 1 light chain 3 A (MAP1LC3A) was used for immunocytochemical examination. The staining intensity of the MAP1LC3A protein and autophagy positivity were lower in the malignant cells; however, they were higher in the NILM-INF and atypical cells. A statistically significant correlation between the malignant and normal cells was obtained for the autophagy positivity (P = 0.012). In view of the staining intensity of MAP1LC3A protein by the H-score method, a significant correlation was found between the NILM-INF and the normal cells (P = 0.015). Autophagy was detected in various cervicovaginal smears for the first time in this study. Our findings indicate that an autophagy process is essential in infectious cells as well as in the transformation of atypical cells into malignant cells in carcinogenesis.

B-cell Follicular Lymphoid Hyperplasia (Pseudolymphoma) of the Oral Cavity: A Diagnostic Quandary.

Pseudolymphoma is a reactive process involving lymphadenopathy, polyclonal proliferation of B or T-cells, simulating oral lymphoma. With its incidence being very rare, only four cases have been reported in oral cavity with the detailed immunocytochemical examination, which can be due to this entity's unawareness, underdiagnosis or overdiagnosis. It is prerogative to perform immunocytochemical investigations to prevent overdiagnosis as lymphoma, which can be debilitating to the patient. Wherein the treatment of pseudolymphoma initially includes topical or intralesional corticosteroid, antibiotics to surgical and radiotherapy based on its etiology. Herein, we discuss B-cell follicular lymphoid hyperplasia previously diagnosed as small round cell tumor.

Возможности цитологического исследования экссудатов серозных полостей в диагностике онкопатологии в условиях многопрофильного стационара

Cytological criteria of tumors in exudate fluids are associated with certain subjective difficulties, one of which is the differential diagnosis of proliferating mesothelial and malignant neoplasm cells. The study aimed to show the possibilities of cytological examination of exudates from serous cavities in the diagnosis of oncopathology. Material and methods. From 2018 to 2021, 10,082 serous cavity effusions (pleural – 8,166 (81%), abdominal cavity – 1,512 (15%), pericardial – 404 (4%)) were included in the cytological examination. Microscopic examination of traditional preparations was carried out, and immunocytochemical (ICC) examination was carried out in difficult diagnostic situations. Results. Analysis of the study showed that by the traditional cytological method in effusion fluids in women, metastatic lesions of the serous cavities were diagnosed in 672 cases (58%), mainly due to the progression of breast cancer (26%). In men, pleurisy was mainly due to metastasis of adenocarcinoma of the lung – 266 cases (23%). ICC research increased the diagnostic accuracy of cytological examination by 62–93% and the specificity – by 95–99%. Conclusions. An algorithm has been developed for conducting ICS studies, differing in the number of monoclonal antibodies used, to determine the histological form of a malignant tumor and the source organ of metastasis. In specific cases, conducting ICR studies with 2–3 monoclonal antibodies may be quite enough to confirm the histological form of the tumor and, where necessary, perform additional ICR studies without significant loss of time for obtaining results.

Usefulness and Limitations of Cryopreservation for Immunocytochemical Staining of Canine Cytological Specimens for Detection of Cytokeratin and Vimentin.

Immunocytochemistry is an advanced diagnostic tool for identifying the origin of tumor cells. This study aimed to highlight the usefulness of cryopreserved, air-dried cytological samples in detecting cytokeratin and vimentin. Air-dried cytological smear samples were prepared from a total of 39 resected canine tumors and stored in a medical freezer without fixation. The duration of cryopreservation ranged from 2 to 56 months. The same tumors were processed for routine histopathological examination. Based on the morphological diagnosis, cryopreserved FNA smears from epithelial tumors were stained by enzymatic immunocytochemistry (ICC) for cytokeratin; those from mesenchymal and melanocytic tumors were stained by ICC for vimentin. To ascertain the positivity of tumor cells to the selected markers, tissue paraffin-embedded sections were also stained by immunohistochemistry (IHC) for the same markers. Immunoreactivity for cytokeratin was detected in cryopreserved cytological smears for a maximum of 46 months. Immunoreactivity for vimentin was clearly detected for 33 months. Smears stored at room temperature for 1 week did not show any signals under immunocytochemical examination. Thus, immunocytochemistry for cytokeratin and vimentin can be safely applied to air-dried smears cryopreserved in a freezer for at least 33 months.

Distinct sets of olfactory receptors highly expressed in different human tissues evaluated by meta-transcriptome analysis: Association of OR10A6 in skin with keratinization.

Olfactory receptors (ORs) are expressed in many tissues and have multiple functions. However, most studies have focused on individual ORs. Here, we aimed to conduct a comprehensive meta-transcriptome analysis of OR gene expression in human tissues by using open-source tools to search a large, publicly available genotype-tissue expression (GTEx) data set. Analysis of RNA-seq data from GTEx revealed that OR expression patterns were tissue-dependent, and we identified distinct sets of ORs that were highly expressed in 12 tissues, involving 97 ORs in total. Among them, OR5P2, OR5P3 and OR10A6 were associated with skin. We further examined the roles of these ORs in skin by performing weighted gene correlation network analysis (WGCNA) and c3net analysis. WGCNA suggested that the three ORs are involved in epidermal differentiation and water-impermeable barrier homeostasis, and OR10A6 showed the largest gene sub-network in the c3net network. Immunocytochemical examination of human skin keratinocytes revealed a sparse expression pattern of OR10A6, suggesting that it is not uniformly distributed among all keratinocytes. An OR10A6 agonist, 3-phenylpropyl propionate (3PPP), transiently increased intracellular Ca2+ concentration and increased cornified envelope (CE) production in cultured keratinocytes. Knock-down of OR10A6 diminished the effect of 3PPP. Overall, integration of meta-transcriptome analysis and functional analysis uncovered distinct expression patterns of ORs in various human tissues, providing basic data for future studies of the biological functions of highly expressed ORs in individual tissues. Our results further suggest that expression of OR10A6 in skin is related to epidermal differentiation, and OR10A6 may be a potential target for modulation of keratinization.

Diagnosis of lung adenocarcinoma using a combination of traditional and liquid cytology with immunocytochemical examination. Clinical case

Diagnosis of lung adenocarcinoma using a combination of traditional and liquid cytology with immunocytochemical examination. Clinical case

STAGES OF CYTOLOGICAL EXAMINATION (USING IMMUNOCYTOCHEMICAL EXAMINATION) OF EFFUSION FLUIDS

Relevance: Cytological criteria of tumors in exudate fluids are associated with specific subjective difficulties, one of which is the differential diagnosis of proliferating mesothelial and adenocarcinoma cells.&#x0D; The study aimed to: increase the informational value of cytological diagnostics in a multidisciplinary hospital.&#x0D; Methods: From 2018 to 2021, 10,082 serous cavity effusions (pleural – 8,166 (81%), abdominal cavity – 1,512 (15%), pericardial – 404 (4%)) were included in the cytological examination. Microscopic examination of traditional preparations was carried out, and immunocytochemical (ICC) examination was carried out in difficult diagnostic situations.&#x0D; Results: In this study, in women, the traditional cytological examination of effusion fluids revealed metastatic lesions of the serous cavities in 672 cases (58%), mainly due to breast cancer progression (26%). In men, pleurisy was primarily due to metastasis of adenocarcinoma of the lung – 266 cases (23%). ICC research increased the diagnostic accuracy of cytological examination by 62-93% and the specificity – by 95-99%.&#x0D; Conclusion: An algorithm for conducting ICH studies, differing in the number of panels of monoclonal antibodies used to determine the histological form and organ — the source of the tumor, has been developed. In specific cases, conducting ICR studies with 2-3 monoclonal antibodies may be enough to confirm the histological form of cancer and, where necessary, perform additional ICR studies without significant loss of time for obtaining results.

International experience of laboratory methods in the cervical cancer screening

Cervical cancer (CC) is a serious health problem all over the world. CC is a fully preventable disease; however, it remains one of the leading causes of death among women with carcinomas. The screening helps to reduce morbidity and mortality. Two tests are used as screening tests in the world: сervical cytology and detection of human papillomavirus (HPV). Cervical cytology (Pap test) is a traditional test of CC screening. Two types of Pap test are used in modern laboratory practice: liquid and traditional methods. HPV testing is now used as the primary screening tool for СС in some countries. However, a key factor in effective screening is the coverage of the population and the organization of the screening. Immunocytochemical examination is an additional method used to improve the accuracy of diagnosis, p16/Ki-67 dual staining. This review focuses on the laboratory methods used in the world practice of screening. The literature search for this review was conducted using PubMed, MedLine and Embase.

Establishment of an in vitro model from the vulnerable fish species Coregonus maraena (maraena whitefish): Optimization of growth conditions and characterization of the cell line.

In this study, a cell line of the fish species Coregonus maraena was produced for the first time. C.maraena is an endangered species, and studies indicate that this fish species will be affected by further population declines due to climate change. This cell line, designated CMAfin1, has been maintained in Leibovitz L-15 supplemented with 10% fetal bovine serum over 3 years. Both subculturing and storage (short-term storage at -80°C and long-term storage in liquid nitrogen) was successful. Cell morphology and growth rate were consistent from passage 10 onwards. Immunocytochemical examination of cellular proteins and matrix components confirmed the mechanical stability of the cells. Actin, fibronectin, vinculin, vimentin, and tubulin are present in the cells and form a network. In addition, the transport of molecules is ensured by the necessary proteins. Gene expression analysis showed a shift in the expressions of stem cell markers between younger and higher passages. While SOX2 and IGF1 were more highly expressed in the seventh passage, SOX9 and IGF2 expressions were significantly increased in higher passages. Therefore, the stable cell culture CMAfin1 can be used for applied analysis to further understand the cell physiology of C.maranea.

Evaluation of P16/Ki67 (CINtecPlus) and L1-capsid compared with HPV-genotyping in cervical cytology in women ≥35 years old focusing on patients with atypical squamous cells of undetermined significance.

Cervical cancer is the third most common cancer in women worldwide. Conventional cytological examination as a screening method with Papanicolaou has been established to reduce the incidence of dysplasia and cervical cancer for years. In addition to the conventional screening, the introduction of immunocytochemical examinations, including CINtecPlus and L1-capsid, has been demonstrated to have a positive impact on screening results. In addition to morphological screening methods, human papillomavirus (HPV)-testing has also been demonstrated to possess an enormous potential in the cervical screening process. Additionally, different screening models ranging from conventional cytological screening to primary HPV-testing do exist in different countries. At the beginning of the year 2020, a combination of cytological screening and HPV-testing was introduced in Germany for women ≥35 years. The aim of the present study was to evaluate the role of morphological screening, including immunocytochemistry, and to compare it with HPV-genotyping. Immunocytochemistry was added to confirm the diagnosis but needs established infrastructure and well-trained personnel. Furthermore, there was a need to establish the HPV-screening method. In the Institute for Pathology and Cytology (Schuettorf, Leer, Germany), 146,800 samples of women (>35 years old) were examined between January 2020 and January 2021. The present study retrospectively analyzed 146,800 samples. Each sample was examined using a conventional cytological technique and HPV-high risk-Test (HPV-HR-Test) with Viper-BD. Immunocytochemistry with CINtecPlus and L1-capsid was added in some cases. A total of 555 cases were cytological diagnosed as atypical squamous cells of undetermined significance (ASC-US; IIp). After performing immunocytochemistry, 79% of cases were suspected to be positive and 1.48% of cases were definitely positive. The HPV-HR-Test was positive in 26.4% of cases. Among cases of ASC-US and HPV-HR-negativity, 33.7% were suspicious of immunocytochemical positivity and 0.5% were definitely positive. Among patients with HPV-16-negativity, 13.6% were patients with highly squamous intraepithelial lesion (HSIL) and 22.7% were patients with low-grade squamous intraepithelial lesion (LSIL) and HSIL. Among patients with HPV-18-negativity, 14.3% were patients with HSIL and 19.5% were patients with LSIL and HSIL. There were 107 cases in this group of cases with negativity of both HPV-16 and HPV-18. After performing the colposcopy and biopsy, there were 6.5% with cervical intra-epithelial neoplasia (CIN) I, 8.4% with CIN II and 5.6% with CIN III. In conclusion, there is still a need for conventional cytological examination and maybe the addition of immunocytochemistry to confirm the diagnosis and to exclude dysplasia of cervical epithelium. The HPV-HR-Test is not enough as a screening method and may be misleading.

Comparison of the Effectiveness of the Cell Block Method with Core Tissue Biopsy for Adequate Diagnosis of Lung Malignant

BACKGROUND: An adequate diagnosis of lung malignancy can be detected through examination of pleural fluid and Transthoracic needle aspiration (TTNA) based on the cell block method and tissue core biopsy. The cell block method is an immunocytochemical examination method that is useful for the adequate diagnosis of lung malignancy.&#x0D; AIM: This study was to compare the effectiveness of the cell block method with tissue core biopsy on pleural fluid cytology and TTNA cytology to establish the diagnosis of lung malignancy.&#x0D; METHODS: This study is a diagnostic test with an analytical cross-sectional approach on 15 respondents suspected of lung cancer at Dr. Moewardi Hospital Surakarta in September 2021. Subjects underwent pleural fluid cytology, TTNA cytology based on the cell block method, and tissue core biopsy examination. Data analysis used the Kappa test and bivariate analysis to calculate the p-value.&#x0D; RESULTS: The results of the combined examination of pleural fluid cytology and TTNA cytology based on the cell block method gave a very good level of conformity to the tissue core biopsy with a Kappa value of 0.857 and a p-value of 0.001. The results of the combined examination of pleural fluid cytology and TTNA cytology with cell block method showed the highest percentage for sensitivity (100%), NPV (100%), and accuracy (93%); while the results of the TTNA cytology examination using the cell block method showed the highest percentage of specificity (100%) and PPV (100%) of the tissue core biopsy method.&#x0D; CONCLUSION: The cell block method is very effective and significant in assessing the results of combined pleural fluid cytology and TTNA cytology against the tissue core biopsy method so that it is useful for the adequate diagnosis of lung malignant

The fluorescent immunocytochemical diagnostics of precancerous diseases and cancer of the oral mucosa.

Purpose of the study improving the diagnosis of precancerous diseases and cancer of the oral mucosa using fluorescent immunocytochemical studies by direct immunofluorescence. A clinical laboratory examination of 111 patients was carried out: 46 patients with squamous cell carcinoma of the oral mucosa, 35 people with precancerous lesions (17 leukoplakia, 18 - oral lichen planus) and 30 healthy people. All patients underwent a traditional cytological examination and an additional immunocytochemical examination by direct immunofluorescence, the expression levels of tumor markers P53, P16 and Ki67 were determined. The data were compared with the results of histological analysis. As a result of the study, it was revealed that in patients with cancer, the expression of oncomarker P53 was four times higher than in patients with precancerous pathology. In 6.52% of cases, co-expression of markers Ki67 and P16 was found. Thus, the advantages of fluorescent immunocytochemical diagnostics were the absence of invasive traumatic intake of the biomaterial in the patient, reduction in the timing of obtaining the result, high sensitivity, and the possibility of remote evaluation of the results. Therefore, that increases the accessibility of the method, and the possibility of using this method for a screening study of population.

Malignant Knee Joint Effusion—A New Dimension of Laboratory Diagnostics

Joint effusions are most frequently caused by osteoarthritis, trauma, an infection process or an autoimmune disease. The development of joint effusion due to a tumor process is rare but should be taken into consideration in the diagnostics. Joint effusions are examined mostly by means of microbiology to rule out or confirm pyogenic synovitis. These standard processes may take up to several days. The article presented here describes a unique case of a 74-year-old female diagnosed with a generalized malignant process according to a cytological-energy analysis and an immunocytochemical examination of a malignant joint effusion caused by femoral condyle metastasis. Other widely-used imaging methods such as X-ray, full-body CT scan and also laboratory examinations confirmed the malignancy and the origin. A cytological-energy analysis and an immunocytochemical examination can expedite the diagnostic process, can outline the processes happening in the joint and can indicate further examinations and subsequent therapy. The use of these laboratory methods appears to be a helpful diagnostic option to obtain additional information about a joint effusion, including the information about an ongoing malignant process. In our case report, they helped to confirm the typing of the tumor within three days, without the need for a metastasis biopsy. In appropriate cases, synovial fluid can play a role in tumor diagnostics.

The feasibility of using immunocytochemical research in the preoperative diagnosis of thyroid tumors

Background. Thyroid cancer has long been an urgent problem of health care in Ukraine and requires both improved timely detection of the disease and increased coverage by special treatment of this category of patients to improve its results. Objective. Investigate the complex of immunocytochemical and ultrasound characteristics of thyroid tumors to improve diagnostic algorithms in the preanalytical stage. Methods. During the the research work the results of diagnostics of 60 patients with thyroid neoplasms aged 21 to 83 years who underwent fine-needle aspiration. Patients were divided into three groups according to the Bethesda System (BSRTC) classification category. The 1st group included 18 patients of category II BSRTC (Benign. Group 2 included 20 patients with category V BSRTC (Suspicious for Malignancy – SFM). And other 22 patients with category VI BSRTC (malignant tumor) formed the 3rd group). Patients of all groups underwent questionnaires, ultrasound examination with FNA, cytological and immunocytochemical examination. Results. There is no clear link between E-cadherin expression and neoplasia. Negative expression occurred in 36.4% of cases of metastatic lesions. This partially satisfies the requirements of timely preoperative diagnosis. That is why E-cadherin cannot be used as a separate marker. The expression of cytokeratin-19 with different degrees of intensity of cytoplasmic staining of tumor cells was determined in all studied neoplasms of V and VI category on the BSRTC scale. Strong expression of cytokeratin-19 was observed in 40.0% of cases with BSRTC-5 and 45.4% with BSRTC-6, moderate - in 50.0% and 36.4% of cases, respectively, weak - in 10.0% and 18.2%, without significant differences between groups (p&gt; 0.05). At the same time, in the benign process in 55.6% of cases there was no reaction of cytokeratin-19, and in 44.4% - weak expression (p &lt;0.001 in comparison with the 2nd and 3rd groups). Conclusion. Benign thyroid disease (category II BSRTC) is likely to be associated with younger patients (46.6 ± 3.7 years), low serum REA (less than 0.95 ng / ml), poor thyroglobulin expression (77.8 %), negative reaction with TTF-1 (100%) and cytokeratin-19 (55.6%). Differential-prognostic markers of thyroid cancer with risk of malignancy (V category BSRTC) include elevated serum REA (0.95 ng / ml and above), the presence of a moderate reaction with antibodies to thyroglobulin (80.0%), a positive reaction - to TTF-1 (100.0%) and E-cadherin (90.0%), with moderate or strong expression of cytokeratin-19 (90.0%).

Comparative evaluation of the results of an immunocytochemical study of P16/Ki-67 coexpression in patients with cervical intraepithelial neoplasia associated with human papillomavirus

Aim. To evaluate the results of an immunocytochemical study of p16/Ki-67 double staining in the cervical epithelium of patients with cervical intraepithelial neoplasia (CIN) associated with high-risk human papillomavirus (HPVhr) in comparison with patients without cervical pathology.&#x0D; Materials and methods. The comparative study included the results of examination of 75 patients divided into 4 groups. Patients with CIN1 comprised the 1st main group (n=21), women with CIN2CIN3 the 2nd main group (n=26), the comparison group consisted of patients with latent HPV infection (n=15) and the control group (n=13). The average age of women with cervical HPV infection was 26.46.13 years. Methods of investigation: liquid cytology, colposcopic, histological examination; methods for determining HPVhr DNA; immunocytochemical examination for determining double staining of p16/Ki-67 markers, statistical analysis.&#x0D; Results. A positive p16/Ki-67 double staining test prevailed among patients with CIN (31.9%) compared to patients without cervical pathology (3.6%) (p=0.003) and correlated with the severity of colposcopic signs (rs=+0.397, p=0.0004). In the 1st main group of patients with verified CIN1 and in the comparison group of patients with latent infection, isolated cases of a positive test of double staining of p16/Ki-67 markers in the epithelium were recorded without significant differences between the groups (9.5 и 6.6%, p0.05). In the 2nd main group of patients with verified CIN2, CIN3, a positive test of double staining of p16 and Ki-67 was observed in every second case, dominating relative to the 1st group, the comparison group and the control group (p=0.003, p=0.005, р=0.001 respectively). In the control group, a negative double staining test was established in all cases.&#x0D; Conclusion. Every second patient with CIN2+ associated with HPVhr has a positive test of double staining of the cervical epithelium, with CIN1 it is observed in 9.5% of cases (p=0.003). Among patients with CIN1, there were no differences in the expression of p16/Ki-67 in epithelial cells compared to women without cervical pathology. The data of the immunocytochemical study of p16/Ki-67 in the cervical epithelium of HPVhr positive patients with CIN should be taken into account when choosing a differentiated management strategy.