Analysis of the Clinical Value of Immunohistochemical Testing in the Pathological Diagnosis of Breast Cancer

Objective:To investigate the clinical value of immunohistochemistry(IHC)detection in the pathological diagnosis of breast cancer.Methods:Eighty breast cancer patients admitted to Baoding No.1 Central Hospital from June 2022 to June 2023 were selected as study subjects.The samples were divided into a positive group(40 cases)and a negative group(40 cases)according to ER and PR test results.Immunohistochemistry was performed on all patients to compare the differences between the two groups in C-erbB-2 positive expression and axillary lymph node metastasis.Results:The positive expression rate of C-erbB-2 in the positive group(35.00%)was significantly lower than that in the negative group(80.00%),with a highly significant difference(P<0.001).The axillary lymph node metastasis rate in the positive group(40.00%)was significantly lower than that in the negative group(75.00%),with a significant difference(P<0.05).Conclusion:Immunohistochemical detection in breast cancer pathology enhances diagnostic accuracy,predicts prognosis,and supports personalized treatment by identifying ER,PR,and C-erbB-2.It is worth being widely adopted in clinical practice.

Ultrastructural and Immunohistochemical Characteristics of Corneal Lenticule Extracted during Correction of Residual Myopia in the Long-Term Period after SMILE

Purpose: To evaluate ultrastructural characteristics of lenticule surface extracted during correction of residual myopia in patients after small-incision lenticule extraction (SMILE). Methods and material: This study had a prospective, consecutive, comparative design. Sixteen patients (16 eyes) underwent additional intervention for residual myopia correction after SMILE. 16 specimens of removed lenticules underwent morphological examination. Markers and reagents were used to determine actin microfilaments, neutral fats and cell nuclei. The tissue was analyzed in layers in 2D slices form, volumetric Z-stacks, or selected areas were formed in orthogonal projections. The surface of the extracted lenticule was analyzed using scanning electron microscopy. Patients’ refractive outcomes were measured postoperatively (1 day;1 and 3 months). Results: Postoperatively uncorrected distance visual acuity (20/20 or better) was in 100% cases 3 months after surgery. Ultrastructural studies have shown the difference in surfaces of the newly formed lenticule. Structural changes of the posterior lenticule surface were characterized by ruptures of collagen fibers on its surface, degenerative changes in keratocytes with signs of colliquation necrosis, cell apoptosis and F-actin in cell cytoplasm. Conclusion: Collagen fibers are immersed in the stroma on the anterior surface of the lenticule. There is no complete structure restoration of collagen fibers explaining the lack of tight adhesion of anterior and posterior surfaces of the intrastromal space even in the long-term postoperative period. There are no degenerative changes of keratocytes on the anterior lenticule surface, that is, their changes in SMILE are reversible in most cases.

Role of FOXP3 in Immunohistochemical Expression in Preeclampsia

Background: FoxP3 gene variants have been linked to endometriosis, infertility, and autoimmune illnesses, according to numerous researches. Maternal sensitivity to the PE gene and the genetic variations of FoxP3 has not been thoroughly investigated. Objective: Investigation of the immune-histochemical expression of FoxP3 in placental tissue of PE patients. Methods: A total of 26 pre-eclamptic women as a case and 26 ethnically matched healthy pregnant women as a control group aged between 18 and 40 years old of different gravidity and parity referred to the labor ward for delivery either by vaginal delivery or cesarean section was enrolled to investigate the immunohistochemical expression of FOXP3 in placental tissue of PE patients. Results: Lower expression of FOXP3 IHC was statistically significant and noted in the group of preeclampsia compared to the healthy control group. Lower gestational age at delivery and a higher percentage of cesarean section were statistically significant and noted in the group of preeclampsia compared to the healthy control group. Conclusion: In comparison to the healthy control group, preeclampsia patients had statistically significantly lower FOXP3 IHC expression, and FOXP3 polymorphism was associated with the development of PE. Our findings can serve as a guide for statistical analyses and functional investigations that are more in-depth.

Immunohistochemical Analysis of IAA in Anthers of the Photoperiod-sensitive Genic Male-sterile Rice

The immunohistochemical localization of IAA and the comparison of their relative levels were carried out for the first time in the anthers of Nongken 58S and its wild type Nongken 58 (Oryza sativa subsp. japonica) after long_day and short_day treatments. The distribution of free_IAA in anthers and its dynamic variation could be reflected by this method. The results showed that the IAA level in the anthers of Nongken 58S after long_day treatment was much lower than that in short_day_treated Nongken 58S and those in wild type Nongken 58 in five stages from pistil and stamen primordia formation to late uninucleate stage. The possible reasons for IAA deficiency in Nongken 58S_LD anthers and its relationship with fertility alteration were also discussed.

Immunohistochemical prognostic markers of esophageal squamous cell carcinoma:a systematic review

Background: Esophageal squamous cell carcinoma(ESCC) is an aggressive malignancy, with a high incidence and poor prognosis. In the past several decades, hundreds of proteins have been reported to be associated with the prognosis of ESCC, but none has been widely accepted to guide clinical care. This study aimed to identify proteins with great potential for predicting prognosis of ESCC.Methods: We conducted a systematic review on immunohistochemical(IHC) prognostic markers of ESCC according to the 2009 Preferred Reporting Items for Systematic Reviews and Meta-Analyses(PRISMA) Guidelines. Literature related to IHC prognostic markers of ESCC were searched from PubMed, Embase, Web of Science, and Cochrane Library until January 30 th, 2017. The risk of bias of these original studies was evaluated using the Quality in Prognosis Studies(QUIPS) tool.Results: We identified 11 emerging IHC markers with reproducible results, including eight markers [epidermal growth factor receptor(EGFR), Cyclin D1, vascular endothelial growth factor(VEGF), Survivin, Podoplanin, Fascin,phosphorylated mammalian target of rapamycin(p-mTOR), and pyruvate kinase M2(PKM2)] indicating unfavorable prognosis and 3 markers(P27, P16, and E-cadherin) indicating favorable prognosis of ESCC.Conclusion: Strong evidence supports that these 11 emerging IHC markers or their combinations may be useful in predicting prognosis and aiding personalized therapy decision-making for ESCC patients.

Epithelioid angiomyolipoma of the liver:Cross-sectional imaging findings of 10 immunohistochemically-verified cases

AIM： To retrospectively evaluate the computed tomography （CT）/magnetic resonance imaging （MRI） imaging features of epithelioid angiomyolipoma of the liver （Epi-HAML）, with pathology as a reference. METHODS： The CT/MRI findings （number, diameter, lobar location, and appearance of lesions） in a series of 10 patients with 12 pathologically proven epithelioid angiomyolipomas of the liver were retrospectively analyzed. The imaging features, including attenuation/ signal intensity characteristics, presence of fat, hypervascular, outer rim, and vessels within lesion, were evaluated and compared with that of non-Epi- HAML in 11 patients （13 lesions）. The Fisher exact test was used to compare difference in probability of imaging features between the two types. RESULTS： For 21 patients, CT images of 15 patients and MR images of six patients were available. No patient underwent two examinations. For the 15 patients with a CT scan, all HAML lesions in the two groups （10 Epi-HAML and seven non-Epi-HAML） manifested as hypoattenuation. For the six patients with MRI, all lesions （two Epi-HAML and six non-Epi- HAML） were hypointense on TlWI （fat suppression） and hyperintense on T2WI. There were 10 non-Epi-HAML, but only two Epi-HAML lesions showed the presence of fat, which significantly different between the two types （P = 0.005）. On the dynamic contrast enhancement （DCE） imaging, eight Epi-HAML, and 13 non-Epi lesions manifested as hypervascular. Punctate or curved vessels were displayed in 10 Epi-HAML as well as in nine non- Epi lesions and outer rim enhancement could be found with eight Epi-HAML as well as six non-Epi lesions. CONCLUSION： Little or no presence of adipose tissue was found to be an imaging feature of Epi- HAML, compared with the non-Epi type. In addition, hypervascularity with opacification of central punctiform or filiform vessels on DCE would be a characteristic enhancement pattern for Epi-HAML.

Management of hepatocellular carcinoma:Predictive value of immunohistochemical markers for postoperative survival

Hepatocellular carcinoma(HCC) accounts for over 90% of all primary liver cancers. With an ever increasing incidence trend year by year,it has become the third most common cause of death from cancer worldwide. Hepatic resection is generally considered to be one of the most effective therapies for HCC patients,however,there is a high risk of recurrence in postoperativeHCC. In clinical practice,there exists an urgent need for valid prognostic markers to identify patients with prognosis,hence the importance of studies on prognostic markers in improving the prediction of HCC prognosis. This review focuses on the most promising immunohistochemical prognostic markers in predicting the postoperative survival of HCC patients.

Hepatocellular Tumors:Immunohistochemical Analyses for Classification and Prognostication

Following the classification of hepatocellular nodules by the International Working Party in 1995 and further elaboration by the International Consensus Group for Hepatocellular Neoplasia in 2009, entities under the spectrum of hepatocellular nodules have been better characterized. Research work hence has been done to answer questions such as distinguishing high-grade dysplastic nodules from early hepatocellular carcinoma (HCC), delineating the tumor cell origin of HCC, identifying its prognostic markers, and subtyping hepatocellular adenomas. As a result, a copious amount of data at immunohistochemical and molecular levels has emerged. A panel of immunohistochemical markers including glypican-3, heat shock protein 70 and glutamine synthetase has been found to be of use in the diagnosis of small, well differentiated hepatocellular tumors and particularly of HCC. The use of liver fatty acid binding protein (L-FABP), β-catenin, glutamine synthetase, serum amyloid protein and C-reactive protein is found to be helpful in the subtyping of hepatocellular adenomas. The role of tissue biomarkers for prognostication in HCC and the use of biomarkers in subclassifying HCC based on tumor cell origin are also discussed.

Evaluation of a Direct Rapid Immunohistochemical Test (dRIT) for Rapid Diagnosis of Rabies in Animals and Humans

Presently the gold standard diagnostic technique for rabies is the direct immunofluorescence assay （dFA） which is very expensive and requires a high level of expertise. There is a need for more economical and user friendly tests, particularly for use in developing countries. We have established one such test called the direct rapid immunohistochemical test （dRIT） for diagnosis of rabies using brain tissue. The test is based on capture of rabies nucleoprotein （N） antigen in brain smears using a cocktail of biotinylated monoclonal antibodies specific for the N protein and color development by streptavidin peroxidase-amino ethyl carbazole and counter staining with haematoxollin. The test was done in parallel with standard FAT dFA using 400 brain samples from different animals and humans. The rabies virus N protein appears under fight microscope as reddish brown particles against a light blue background. There was 100 % correlation between the results obtained by the two tests. Also, interpretation of results by dRIT was easier and only required a light microscope. To conclude, this newly developed dRIT technique promises to be a simple, cost effective diagnostic tool for rabies and will have applicability in field conditions prevalent in developing countries.

Automation of immunohistochemical evaluation in breast cancer using image analysis

AIM:To automate breast cancer diagnosis and to study the inter-observer and intra-observer variations in the manual evaluations.METHODS:Breast tissue specimens from sixty cases were stained separately for estrogen receptor(ER),progesterone receptor(PR)and human epidermal growth factor receptor-2(HER-2/neu).All cases were assessed by manual grading as well as image analysis.The manual grading was performed by an experienced expert pathologist.To study inter-observer and intra-observer variations,we obtained readings from another pathologist as the second observer from a different laboratory who has a little less experience than the first observer.We also took a second reading from the second observer to study intra-observer variations.Image analysis was carried out using in-house developed software(TissueQuant).A comparison of the results from image analysis and manual scoring of ER,PR and HER-2/neu was also carried out.RESULTS:The performance of the automated analysis in the case of ER,PR and HER-2/neu expressions was compared with the manual evaluations.The performance of the automated system was found to correlate well with the manual evaluations.The inter-observer variations were measured using Spearman correlation coefficient r and 95%confidence interval.In the case of ER expression,Spearman correlation r=0.53,in the case of PR expression,r=0.63,and in the case of HER-2/neu expression,r=0.68.Similarly,intra-observer variations were also measured.In the case of ER,PR and HER-2/neu expressions,r=0.46,0.66 and 0.70,respectively.CONCLUSION:The automation of breast cancer diagnosis from immunohistochemically stained specimens is very useful for providing objective and repeatable evaluations.

Does immunohistochemical staining have a clinical impact in early gastric cancer conducted endoscopic submucosal dissection?

AIM: To evaluate clinicopathologic parameters and the clinical significance related lymphovascular invasion (LVI) by immunohistochemical staining (IHCS) in endoscopic submucosal dissection (ESD). METHODS: Between May 2005 and May 2010, a total of 348 lesions from 321 patients (mean age 63 ± 10 years, men 74.6%) with early gastric cancer (EGC) who met indication criteria after ESD were analyzed retrospectively. The 348 lesions were divided into the absolute (n = 100, differentiated mucosal cancer without ulcer ≤ 20 mm) and expanded (n = 248) indica-tion groups after ESD. The 248 lesions were divided into four subgroups according to the expanded ESD indication. The presence of LVI was determined by factor Ⅷ-related antigen and D2-40 assessment. We compared LVI IHCS-negative group with LVI IHCSpositive in each group. RESULTS: LVI by hematoxylin-eosin staining (HES) and IHCS were all negative in the absolute group, while was observed in only the expanded groups. The positive rate of LVI by IHCS was higher than that of LVI by HES (n = 1, 0.4% vs n = 11, 4.4%, P = 0.044). LVI IHCS-positivity was observed when the cancer invaded to the mucosa 3 (M3) or submucosa 1 (SM1) levels, with a predominance of 63.6% in the subgroup that included only SM1 cancer (P < 0.01). In a univariate analysis, M3 or SM1 invasion by the tumor was significantly associated with a higher rate of LVI by IHCS, but no factor was significant in a multivariate analysis. There were no cases of tumor recurrence or metastasis during the median 26 mo follow-up. CONCLUSION: EGCs of the absolute group are immunohistochemically stable. The presence of LVI may be carefully examined by IHCS in an ESD expanded indication group with an invasion depth of M3 or greater.

DIAGNOSTIC IMPLICATIONS OF IMMUNOHISTOCHEMICAL MARKERS IN UTERINE SMOOTH MUSCLE TUMORS

To evaluate the diagnostic implications of immunohistochemical markers in uterine smooth muscle tumors. Methods: Formalin-fixed paraffin-embedded tissue blocks were selected from 17 uterine leiomyosarcomas, 40 uterine unusual leiomyomas and 25 uterine usual leiomyomas. Utilizing immunohistochemical techniques with antigen retrieval, serial sections of each tumor for immunoreactivity with myogenic markers, ovarian steroid receptors, CD44v3, proliferating cell nuclear antigen and mast cells were assessed. Results: Although the myogenic markers and CD44v3 showed less frequent positivity in uterine leiomyosarcomas than those in unusual leiomyomas, they were not reliable markers for differentiating leiomyosarcoma from leiomyoma. Uterine leiomyosarcoma tended to have lower ovarian steroid receptors immunoreactivity rates than leiomyoma. Leiomyoma tended to have a higher quantity of intratumoral mast cells than leiomyosarcoma, while the expression of proliferating cell nuclear antigen was lower in them. Conclusion: Because the estimation of mitotic count was subject to significant variation, the immunohistochemical expression of ovarian steroid receptors, mast cells and proliferating cell nuclear antigen seemed to be helpful for the discrimination of unusual leiomyoma from leiomyosarcoma.

IMMUNOHISTOCHEMICAL STUDY ON S100 PROTEIN-POSITIVE DENDRITIC CELLS IN PATIENTS WITH GASTRIC CARCINOMA

The density of dendritic cells (DC) and macro-phages (Mφ) in tissue specimens of gastric carcinoma (GC n=65) was investigated by ABC im-munohistochemical method using anti-S100 protein and anti-lysozyme antibodies, and was compared with that in gastric ulcer (GU n=19), chronic atrophic gastritis (CAG n=28) and normal gastric mucosa (NGM n=15). The mean density if DC (cells/mm2) in GC (15.0 was significantly higher than that in NGM (3.8) and GU (8.3), but was remarkably lower when compared to that in CAG (29.5) (P<0.01). Statistically significant difference in the population density of DC was observed between well- and poorly-differentiated GC (P<0.01). With their unique dendritic processes, DC were mainly concentrated within dense lymphoid infiltrates or in the T-area of reactive lymphoid follicles and were interspersed among the tumor cells. In contrast, Mφ were present around the necrotic foci and were rarely seen within the non-necrotic neoplastic tissues. These data suggest that DC, which differ in morphology, distribution, number and function form Mφ may be more directly involved in the host immune reaction against tumor by acting as antigen presenting cells.

PROLIFERATION RATE OF COLONIC NEOPLASMS AND MUCOSA ADJACENT TO NEOPLASMS: IMMUNOHISTOCHEMICAL STUDY

Biopsy specimens of normal mucosa (n=5). adenomas (n -13), adenocarcinomas (n = 8). mucosa adjacent to adenoma (n=10) mucosa adjacent to adenocarcinoma (n = 7 ) at the large bowel were investigated by an iminunohistochemical method using 5- bromodeoxynldine (BrdUrd) . The labeling index (LI) was significantly lower in normal nucosa than mucosa adjacent to neoplasms and adenomas and adenocarcinomas. The proliferative zone was confined to the lower two- third at the crypt in normal mucosa and in mucosa adjacent to neoplasms. The labeled cells were either present in the upper third or scattered along the crust and in surface epithelium. The results support the adenmo-carcinoma sequence.

Immunohistochemical Localization of Aspergillus and p53 in Human Lung Tissues

Aspergilli are filamentous fungi which can cause opportunistic infections in Acquired Immunodeficiency Syndrome (AIDS) patients. Aspergilli can be found in human tissues either in the form of spores or hyphae. p53 is a tumor suppressor gene located in the short arm of chromosome 17. It is a potent transcriptional regulator of genes which are involved in many cellular activities including cell cycle arrest, apoptosis and angiogenesis. A loss of tumor suppressor function of p53 is the most common event leading to the development of human cancers. The rate by which p53 has a homology between different species has been reported from human to other vertebrates, it has been reported that it is available within Drosophila melanogaster and C. elegans [1] [2]. The aim of this study is to check if p53 is localized within Aspergilli or not using immunohistochemical techniques and study the relationship between Aspergilli infection and p53 in human lung tissues. 45 different samples of lung tissues, diagnosed as being none tumor, were taken randomly during the year of 2003-2004 from the autopsy cases submitted to the forensic medicine center in Irbid, Jordan. The sample group consisted of 12 females and 33 males. Labeled Streptavidin Biotin (LSAB) method and Mach-4 method were used to determine the Aspergilli infection and p53. The results show that the Aspergillus is presented in all used samples (100% of the infection) in the form of spores or hyphae and all infected samples have mutant p53 molecules (p53 was located in Aspergillus spores and hyphae). According to this study, it is safe to posit that the mutant p53 molecules may be used by Aspergillus for its multiplication. Seemingly it is a biological behavior of Aspergillus to produce p53. The fate of the p53 is questioned, is it going to interact with the human cells initiating cancer? Further experimental investigations are required to determine such pathway. In conclusion, this study shows that Aspergillus is a producing agent for p53 and Aspergillus pathogenicity is caused by production of p53.

IMMUNOHISTOCHEMICAL STUDY OF ALPHA-1-ANTICHYMOTRYPSIN IN GLIOMA

GFAP is a specific antigen of glial element, but Alpha-1-antichymotrypsin has not been reported in the literature. Alpha-1-antichymotrypsin was guided by GFAP using PAP method to the astrocytes of 137 gliomas. 120 (87%) gliomas were positive for Alpha-1-antichymotrypsin. Of these 120 gliomas, 86 (72%) gave diffuse distribution, 17 (14%) gave focal distribution, and 17 (14%) gave scattered distributions. Alpha-1-antichymotrypsin in glioma tissue may be an important tumor marker for diagnosis.

HISTOCHEMICAL AND IMMUNOHISTOCHEMICAL STUDY OF EXTRAMAMMARY PACET'S DISEASE

The histogensis of extramammary Paget's disease has not been solved and remained controversial. Eight cases of extramammary Paget's disease of geni-tocrural region were investigated by alcian blue and PAS stain and immunoreaction of anti-CEA and anti-keratin. It was found that the pattern and intensity of alcian blue, PAS staining were identical for Paget cells and secretory cells of apocrine sweat gland; and CEA immunoreactivity was uniformly observed in both Paget cells and eccrine sweat gland. The keratin immunoreaction was positive in keratinocytes, apocrine and eccrine sweat gland, whereas Paget cells were negative. These results suggested that Paget cells of extramammary Paget's disease could be derived from multipotential epidermal germ cells.

IMMUNOHISTOCHEMICAL STUDY OF CARCINOEMBRYONIC ANTIGEN IN THE TRANSITIONAL MUCOSA ADJACENT TO COLORECTAL CANCER

A combined histopathological, mucin histochemi-cal and immunohistochemical study of the transitional mucosa (TM) adjacent to colorectal cancer is presented. Twenty-six resected specimens were studied by hematoxylin and eosin (HE) and high iron diamine-alcian blue (HID-AB). Carcinoem-bryonic antigen (CEA) was demonstrated by peroxi-dase antiperoxidass (PAP) technique. The appearance of the TM is usually thicker, longer and dilated crypts with increased immature and intermediate cells. Variable amount of sialomucins and decrease sulphomucins content as well as increased CEA content are found in the TM. These changes are not seen in non-transitional zone and normal colorectal mucosa. It is suggested that the mucin changes and expression of CEA in the TM may indicate an early primary premalignant changes and may be one of the reasons for the TM affecting the prognosis of patients with large bowel cancer after radical resection.

IMMUNOHISTOCHEMICAL STUDY OF SIXTEEN CASES OF CHORDOMA

Sixteen cases of chordoms were studied with various polycolonal and monoilonal antibodies and ABC technic.such as antibodies S-100 protein, neurou specific enodas(NSE ), lysozyme, α1- AT, Myoglobin, desmin, keratin and cytokeratin. Two embryonic notochords and three chondrosarcomas were studied as control. Most of chordomas revealed positive results with NSE ( 12/16 ). Des ( 12 / 16 ) . α1-AT ( 11/ 16 ), MG( 11/16 ) . S-100( 10 /16)and CK ( 8 / 16 ), while K and Lys were negative in all( 16 )cases. It denotes that chordoma has the potentiality for both mesodermal and ectodermal differentiation. Lys was positive in all 3 chondrosarcomas while the CK was negative. So that Lys can be considered as an useful antibody to distinguish chordoma from chondrosarcoma, and CK is somewhat useful for this differential diagnosis too.

Histopathological and Immunohistochemical Study of the Distinction between Oral Lichen Planus and Oral Lichenoid Lesions

Background: Oral potentially malignant disorders, which include oral lichen planus (OLP), are clinical presentations that carry a risk of development to cancer in the oral cavity. Oral lichenoid lesions (OLLs) are also termed interface/lichenoid mucositis. Malignant transformation of them remains controversial, but distinct clinical and histological criteria for how to differentiate OLP from OLLs have not been developed. Objectives: The purpose of this study was to elucidate findings that can allow histopathological differentiation of OLP and OLLs using histomorphological and immunohistochemical analyses. Materials and Methods: Analyses were performed in 10 cases diagnosed with OLP and 9 cases diagnosed with OLLs. Cytokeratin 19 (CK19), Ki-67 and CD3 were used as primary antibodies to detect basal cells, proliferative activity and T-cell distribution, respectively, and Perlecan and COX-2 to evaluate epithelial intracellular arrangements and interstitial distributions of proteoglycans and enzymes. Results: For CK19, positive cells were significantly found in OLLs at both the prominent area and site adjacent to the lesion comparison with those of OLP’s. The number of COX-2 positive cells was significantly higher in spinous and basal layers in OLLs of the prominent area. Additionally, OLLs showed mild to moderate expression for perlecan in the basal to spinous layers and in subepithelial tissue. Conclusion: Almost no basal cells were noted in the prominent area in OLP. COX-2 and perlecan were found in the basal to spinous layers in OLLs. Although there are restrictions, these suggested the possibility of helping to distinguish between OLP and OLLs.