Humans ; Biomarkers, Tumor/metabolism* ; Claudins/analysis* ; Consensus ; Immunohistochemistry/methods* ; Stomach Neoplasms/diagnosis*

OBJECTIVES: Shear wave elastography (SWE) is a novel quantitative elastography technique that can assess the hardness of different tissues. This study introduces a novel shear wave parameter-frequency of mass characteristic (f_mass)-and investigates its correlation, along with other shear wave parameters, with the histopathological features and immunohistochemical (IHC) biomarkers of invasive breast cancer (IBC). The study aims to explore whether SWE can provide useful information for IBC treatment and prognosis. METHODS: With the pathological results as the gold standard, 258 malignant breast lesions were collected, and all patients underwent conventional ultrasound and SWE examinations. The SWE parameters [maximum elastic value (E_max), minimum elastic value (E_min), mean elastic value (E_mean), standard deviation of elastic value of the whole lesion (E_sd)] and f_mass] in the transverse and longitudinal orthogonal sections were measured, and their correlations with the prognostic factors of IBC [including tumor diameters, axillary lymph node (ALN) metastasis, lymphatic vessel invasion (LVI), calcification, histological type, histological grade, and IHC biomarkers (ER, PR, HER-2, Ki-67), and molecular subtypes] were analyzed. The correlations between the SWE parameters of the transverse and longitudinal sections of the tumors with different prognostic factors and the above indicators were analyzed. At the same time, the receiver operating characteristic (ROC) curve was used to analyze the efficacy of f_mass in predicting ER and PR expression. RESULTS: E_mean, E_max, E_sd, and f_mass were correlated with tumor diameters; E_mean, E_max and E_sd were correlated with histological types and histological grades. E_max and E_sd were correlated with ALN metastasis, LVI and pathological types. In the IHC biomarker-labeled masses, f_mass was correlated with ER and PR (both P<0.05), and E_mean, E_max, and E_sd were correlated with HER-2 and Ki-67 (all P<0.05). E_mean, E_max, and f_mass were all correlated with breast cancer subtypes (all P<0.05), and E_mean and E_max were higher in Luminal B [HER-2(+)] breast cancer, while f_mass was lower in HER-2(+) and triple-negative breast cancer. Among the statistically significant prognostic factors, the P values of the transverse sections of the masses were all less than or equal to those of the longitudinal sections. The AUC of f_mass in the transverse sections of the masses for predicting ER and PR expression were 0.73 (95% CI 0.65 to 0.80) and 0.67 (95% CI 0.60 to 0.74), respectively, with the optimal cut-off values being 76.50 and 60.66, the sensitivities being 72.45% and 81.98%, the specificities being 66.13% and 45.35%, and the accuracies being 70.93% and 69.77%, respectively. The AUC of f_mass in the longitudinal sections of the masses for predicting ER and PR expression were 0.74 (95% CI 0.67 to 0.81) and 0.65 (95% CI 0.58 to 0.72), respectively, with the optimal cut-off values being 131.8 and 137.5, the sensitivities being 69.90% and 66.28%, the specificities being 72.58% and 60.47%, and the accuracies being 70.54% and 64.34%, respectively. The f_mass in the transverse sections of the masses was more statistically significant. CONCLUSIONS The poor prognosis factors of IBC are related to high E_mean, E_min, E_max, E_sd, and low f_mass. The f_mass can predict the expression of ER and PR, and the transverse cut data are more meaningful. SWE is helpful for predicting the invasiveness of IBC.
Humans ; Breast Neoplasms/metabolism* ; Female ; Elasticity Imaging Techniques/methods* ; Biomarkers, Tumor/metabolism* ; Middle Aged ; Adult ; Prognosis ; Immunohistochemistry ; Neoplasm Invasiveness ; Receptor, ErbB-2/metabolism* ; Aged ; Lymphatic Metastasis ; Receptors, Estrogen/metabolism* ; Receptors, Progesterone/metabolism* ; Ki-67 Antigen/metabolism*

In forensic traumatic pathology practice, immunohistochemistry and special staining technique play an important role in wound age estimation and complications of traumatic complication identification. They even play an important role in the identification of special cases, such as snakebites and insulin killings. This article reviews the application and value of immunohistochemistry and special staining techniques in forensic traumatic pathology based on the cases of forensic practice reported in literature.
Forensic Medicine ; Forensic Pathology/methods* ; Immunohistochemistry ; Staining and Labeling

METHODS: For primary human cavernous EC culture, cavernous tissues were implanted into Matrigel in dishes. For primary human cavernous pericyte culture, cavernous tissues were settled by gravity into dishes. We performed immunocytochemistry and Western blot to determine phenotype and morphologic changes from passage 1 to 5. The primary cultured cells were exposed to a normal-glucose (5 mmol/L) or a high-glucose (30 mmol/L) condition, and then tube formation assay was done.RESULTS: We successfully isolated high-purity EC and pericytes from human corpus cavernosum tissue. Primary cultured EC showed highly positive staining for von Willebrand factor, and pericyte revealed positive staining for NG2 and platelet-derived growth factor receptor-β. Primary cultured EC and pericytes maintained their cellular characteristics up to passage 2 or 3. However, we observed significant changes in their typical phenotype from the passage 4 and morphological characteristics from the passage 3. Human cavernous EC or pericytes formed well-organized capillary-like structures in normal-glucose condition, whereas severely impaired tube formation was detected in high-glucose condition.CONCLUSIONS: This study provides a simple and nonenzymatic method for primary culture of human cavernous EC and pericytes. Our study will aid us to understand the pathophysiology of diabetic erectile dysfunction, and also be a valuable tool for determining the efficacy of candidate therapeutic targets.]]>
Blotting, Western ; Cells, Cultured ; Diabetes Mellitus ; Endothelial Cells ; Erectile Dysfunction ; Gravitation ; Humans ; Immunohistochemistry ; Male ; Methods ; Pericytes ; Phenotype ; Platelet-Derived Growth Factor ; von Willebrand Factor

immunohistochemistry (IHC)—22C3, SP263, and SP142. The aim of this study is to evaluate the correlation among the three methods of PD-L1 IHC in non-small cell lung cancer (NSCLC) and clinical significance of PD-L1 expression in lung adenocarcinoma with an epidermal growth factor receptor (EGFR)–tyrosine kinase domain mutation.METHODS: The results of 230 patients who were pathologically confirmed as having NSCLC; tested using PD-L1 IHC 22C3, SP263, and SP142 methods; and evaluated via the peptide nucleic acid clamping method to confirm EGFR mutation, were analyzed in this study.RESULTS: 164 patients underwent both the SP263 and 22C3 tests. There was a significant positive correlation between the outcomes of the two tests (Spearman correlation coefficient=0.912, p<0.001), with a derived regression equation as follows: 22C3=15.2+0.884×SP263 (R2=0.792, p<0.001). There was no relationship between the expression of PD-L1 and clinical parameters, including EGFR–tyrosine kinase inhibitor (TKI) mutation. The PD-L1 expression in patients treated with EGFR-TKI yielded a 2-month-shorter progression period than that in the PD-L1–negative group. However, this did not reach statistical significance (PD-L1<1% vs. PD-L1≥1%, 10 months vs. 8 months).CONCLUSION: The results of the 22C3 and those of SP263 methods were in good correlation with one another. Since the PD-L1 expression is not influenced by the EGFR mutation, it is necessary to perform a PD-L1 test to set the treatment direction in the patients with EGFR-mutant NSCLC.]]>
Adenocarcinoma ; Carcinoma, Non-Small-Cell Lung ; Constriction ; Humans ; Immunohistochemistry ; Lung ; Methods ; Phosphotransferases ; Receptor, Epidermal Growth Factor

PURPOSE: Trastuzumab is an effective treatment for human epidermal growth factor receptor 2 (HER2)-amplified breast cancers. We sought to develop a simple protocol for HER2 image analysis of breast cancer specimens. MATERIALS AND METHODS: In a preliminary test, we found that at least 1000 tumor cells need to be examined in the most strongly stained areas. Next, we evaluated the clinical usefulness of this established protocol of image analysis in 555 breast cancer patients. Results of the HER2 immunohistochemical (IHC) staining were compared between manual scoring and image analysis. RESULTS: The HER2 IHC results obtained by the image analysis method correlated well with those obtained by the manual scoring method (Cohen's kappa=0.830). Using the HER2 silver in situ hybridization (SISH) results as a gold standard, sensitivity values were 72.1% for manual scoring and 74.0% for image analysis; specificity values were 96.2% for manual scoring and 94.7% for image analysis; and accuracy values were 91.7% for manual scoring and 90.8% for image analysis. McNemar's test was applied to the results, and there were no statistically significant differences in sensitivity and specificity between the positive (p=0.688) and negative (p=0.118) SISH groups. CONCLUSION: HER2 image analysis results were similar to those obtained via the manual scoring method, indicating that the use of image analysis can reduce assessment time and effort. We suggest that image analysis-based evaluation of 1000 tumor cells in the most strongly IHC-stained area, regardless of stroma content, is sufficient for determining HER2 expression levels in breast cancer specimens.
Breast Neoplasms ; Breast ; Humans ; Immunohistochemistry ; In Situ Hybridization ; Methods ; Receptor, Epidermal Growth Factor ; Research Design ; Sensitivity and Specificity ; Silver ; Trastuzumab

The locus coeruleus (LC) has been studied in major depressive disorder (MDD) and bipolar disorder (BD). A major problem of immunocytochemical studies in the human LC is interference with the staining of the immunocytochemical end-product by the omnipresent natural brown pigment neuromelanin. Here, we used a multispectral method to untangle the two colors: blue immunocytochemical staining and brown neuromelanin. We found significantly increased tyrosine hydroxylase (TH) in the LC of MDD patients-thus validating the method-but not in BD patients, and we did not find significant changes in the receptor tyrosine-protein kinase ErbB4 in the LC in MDD or BD patients. We observed clear co-localization of ErbB4, TH, and neuromelanin in the LC neurons. The different stress-related molecular changes in the LC may contribute to the different clinical symptoms in MDD and BD.
Aged ; Aged, 80 and over ; Bipolar Disorder ; metabolism ; pathology ; Depressive Disorder, Major ; metabolism ; pathology ; Female ; Humans ; Image Processing, Computer-Assisted ; Immunohistochemistry ; methods ; Locus Coeruleus ; metabolism ; pathology ; Male ; Melanins ; metabolism ; Microscopy ; methods ; Middle Aged ; Neurons ; metabolism ; pathology ; Receptor, ErbB-4 ; metabolism ; Sensitivity and Specificity ; Spectrum Analysis ; methods ; Tyrosine 3-Monooxygenase ; metabolism

Sparse labeling of neurons contributes to uncovering their morphology, and rapid expression of a fluorescent protein reduces the experiment range. To achieve the goal of rapid and sparse labeling of neurons in vivo, we established a rapid method for depicting the fine structure of neurons at 24 h post-infection based on a mutant virus-like particle of Semliki Forest virus. Approximately 0.014 fluorescent focus-forming units of the mutant virus-like particle transferred enhanced green fluorescent protein into neurons in vivo, and its affinity for neurons in vivo was stronger than for neurons in vitro and BHK21 (baby hamster kidney) cells. Collectively, the mutant virus-like particle provides a robust and convenient way to reveal the fine structure of neurons and is expected to be a helper virus for combining with other tools to determine their connectivity. Our work adds a new tool to the approaches for rapid and sparse labeling of neurons in vivo.
Animals ; Cells, Cultured ; Gene Expression ; Genetic Vectors ; genetics ; metabolism ; Green Fluorescent Proteins ; genetics ; metabolism ; Immunohistochemistry ; methods ; Male ; Mice, Inbred C57BL ; Microscopy, Fluorescence ; methods ; Neurons ; cytology ; metabolism ; Purkinje Cells ; cytology ; metabolism ; Semliki forest virus ; genetics

PD-1/PD-L1 inhibitors play an important role in the first-line and second-line treatment of non-small cell lung cancer (NSCLC), indicating a new treatment strategy of NSCLC. Completed clinical trials have shown that effective detection of PD-L1 expression is the key to the use of immunosuppressive agents. However, the gold standard for PD-L1 detection has still lacked. In recent years, immunohistochemistry (IHC) and enzyme-linked immunosorbent assay (ELISA) have been continuously innovated, which accounts for good prospect in PD-L1 detection. The research progress of PD-L1 detection methods in NSCLC is summarized in this review.
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B7-H1 Antigen ; analysis ; Biomarkers, Tumor ; analysis ; Carcinoma, Non-Small-Cell Lung ; diagnosis ; metabolism ; Enzyme-Linked Immunosorbent Assay ; methods ; Humans ; Immunohistochemistry ; methods ; Lung Neoplasms ; diagnosis ; metabolism ; Reproducibility of Results ; Sensitivity and Specificity

PURPOSE: We aimed to analyze the discordance between immunohistochemistry (IHC)-based surrogate subtyping and PAM50 intrinsic subtypes and to assess overall survival (OS) according to discordance. MATERIALS AND METHODS: A total of 607 patients were analyzed. Hormone receptor (HR) expression was evaluated by IHC, and human epidermal growth factor receptor 2 (HER2) expression was analyzed by IHC and/or fluorescence in situ hybridization. PAM50 intrinsic subtypes were determined according to 50 cancer genes using the NanoString nCounter Analysis System. We matched concordant tumor as luminal A and HR+/HER2–, luminal B and HR+/HER2+, HR–/HER2+ and HER2–enriched, and triple-negative breast cancer (TNBC) and normal- or basal-like. We used Ion Ampliseq Cancer Panel v2 was used to identify the genomic alteration related with discordance. The Kaplan-Meier method was used to estimate OS. RESULTS: In total, 233 patients (38.4%) were discordant between IHC-based subtype and PAM50 intrinsic subtype. Using targeted sequencing, we detected somatic mutation–related discordant breast cancer including the VHL gene in the HR+/HER2– group (31% in concordant group, 0% in discordant group, p=0.03) and the IDH and RET genes (7% vs. 12%, p=0.02 and 0% vs. 25%, p=0.02, respectively) in the TNBC group. Among the luminal A/B patients with a discordant result had significantly worse OS (median OS, 73.6 months vs. not reached; p < 0.001), and among the patients with HR positivity, the basal-like group as determined by PAM50 showed significantly inferior OS compared to other intrinsic subtypes (5-year OS rate, 92.2% vs. 75.6%; p=0.01). CONCLUSION: A substantial portion of patients showed discrepancy between IHC subtype and PAM50 intrinsic subtype in our study. The survival analysis demonstrated that current IHC-based classification could mislead the treatment and result in poor outcome. Current guidelines for IHC might be updated accordingly.
Breast Neoplasms ; Breast ; Classification ; Fluorescence ; Genes, Neoplasm ; Humans ; Immunohistochemistry ; In Situ Hybridization ; Methods ; Phenobarbital ; Receptor, Epidermal Growth Factor ; Triple Negative Breast Neoplasms