The clinicopathological impact of tumor-associated macrophages in patients with cutaneous malignant melanoma.

BACKGROUND: Tumor-associated macrophages (TAMs) are an immune component of the cutaneous malignant melanoma (CMM) microenvironment and affect tumor growth. TAMs can polarize into different phenotypes, that is, proinflammatory M1 and anti-inflammatory M2 macrophages. However, the role of the macrophage phenotype in CMM remains unclear. METHODS: We examined 88 patients with CMM. Tissue microarrays were constructed, and the density of M1 and M2 macrophages was analyzed by immunohistochemistry. Immune cells coexpressing CD68 and phosphorylated signal transducer and activator of transcription 1 (pSTAT1) were considered M1 macrophages, whereas those coexpressing CD68 and c-macrophage activating factor (c-Maf) were defined as M2 macrophages. These TAMs were counted, and the relationships between the density of M1 and M2 macrophages and clinicopathological factors including prognosis were investigated. RESULTS: The CD68/c-Maf score ranged from 0 to 34 (median: 5.5). The patients were divided based on the median score into the CD68/c-Maf high (≥5.5) and low (<5.5) expression groups. Univariate and multivariate analyses revealed that CD68/c-Maf expression was an independent predictive factor for progression-free survival and an independent prognostic factor for overall survival. CD68/pSTAT1 expression was found in only two patients. CONCLUSION: We suggest that CD68/pSTAT1 coexpression is rarely observed in patients with CMM, and high CD68/c-Maf expression is a predictor of worse prognosis in these patients.

A genome-wide study of gastric intramucosal neoplasia based on somatic copy number alterations, gene mutations, and mRNA expression patterns.

We performed comprehensive analyses of somatic copy number alterations (SCNAs) and gene expression profiles of gastric intramucosal neoplasia (IMN) using array-based methods in 97 intestinal-type IMNs, including 39 low-grade dysplasias (LGDs), 37 high-grade dysplasias (HGDs), and 26 intramucosal carcinomas (IMCs) with stromal invasion of the lamina propria to identify the molecular mechanism of IMN. In addition, we examined gene mutations using gene panel analyses. We used cluster analyses for exclusion of arbitrariness to identify SCNA patterns and expression profiles. IMNs were classified into two distinct subgroups (subgroups 1 and 2) based on SCNA patterns. Subgroup 1 showed a genomic stable pattern due to the low frequency of SCNAs, whereas subgroup 2 exhibited a chromosomal instability pattern due to the high frequencies of SCNAs and TP53 mutations. Interestingly, although the frequencies of LGD and HGD were significantly higher in subgroup 1 than in subgroup 2, IMC was commonly found in both types. Although the expression profiles of specific mRNAs could be used to categorise subgroups 1 and 2, no clinicopathological findings correlated with either subgroup. We examined signalling pathways specific to subgroups 1 and 2 to identify the association of each subgroup with signalling pathways based on gene ontology tree visualisation: subgroups 1 and 2 were associated with haem metabolism and chromosomal instability, respectively. These findings reveal a comprehensive genomic landscape that highlights the molecular complexity of IMNs and provide a road map to facilitate our understanding of gastric IMNs.

Prognostic and predictive value of CD163 expression and the CD163/CD68 expression ratio for response to adjuvant chemotherapy in patients with surgically resected lung squamous cell carcinoma.

BACKGROUND: Macrophages infiltrating the tumor microenvironment are defined as tumor-associated macrophages (TAMs). TAMs can be polarized into different phenotypes, that is, proinflammatory M1 macrophages or anti-inflammatory M2 macrophages. Particularly, M2 macrophages promote angiogenesis, wound healing, and tumor growth. This study aimed to evaluate whether M2 TAMs can serve as a useful marker to predict prognosis and benefit from adjuvant chemotherapy in patients with surgically resected lung squamous cell carcinomas (SCCs). METHODS: We examined 104 patients with SCC. Tissue microarrays were constructed, and the density of TAMs was analyzed by immunohistochemistry for expression of CD68 and CD163. The relationship between CD68 and CD163 expression and the CD163/CD68 expression rate and clinicopathological characteristics including patient outcomes were investigated. In addition, propensity score matching (PSM) analysis was conducted to test the hypothesis that these cells significantly influenced chemotherapy responses. RESULTS: Univariate analysis revealed that pathological stage, CD163 expression, and the CD163/CD68 expression ratio were significant prognostic factors. Multivariate analysis showed that these factors were all independent prognostic factors. Thirty-four pairs were determined by using PSM analysis. Patients with a low CD163/CD68 expression ratio benefited more from adjuvant chemotherapy than those with a high ratio. CONCLUSION: We suggest that M2 TAMs may be a useful marker to predict prognosis and differential benefit from adjuvant chemotherapy in patients with surgically resected lung SCCs.

Bone formation in beta-tricalcium phosphate-filled bone defects of the rat femur: morphometric analysis and expression of bone related protein mRNA.

The purpose of the current study was to evaluate the bone formation when beta-tricalcium phosphate (TCP) was implanted in bone defects of rat femurs. beta-TCP granules were applied to defects created in the femurs of 65 male rats who were sacrificed 3, 7, 10, 14 or 30 days later. Bone tissues were embedded in paraffin, serial sections were cut and then stained with hematoxylin-eosin. Histomorphometric analyses were also conducted. Furthermore, total mRNAs were extracted, homogenized, and reverse transcribed, after which quantitative PCR assays were conducted with a LightCycler using the double-stranded DNA dye Syber Green I with primers for either rat osteopontin or osteocalcin. Tissues in defects without beta-TCP were used as controls. The amount of newly formed bone tissue in the beta-TCP implanted group was significantly greater in both the side areas and the central area of defects than in the control group. Expressions of osteopontin and osteocalcin mRNAs of cells in the defects of the experimental group were up-regulated compared with the control group at all time periods. Taken together, these results prove that beta-TCP is an appropriate material for osteoconduction and promotes bone formation in bone defects.

Intravascular papillary endothelial hyperplasia arising from the upper lip.

We report a rare case of intravascular papillary endothelial hyperplasia (IPEH) arising from the upper lip. Pathologically, it consisted of a few lobulated masses lined by an incomplete fibrous capsule of variable thickness which was separated from the surrounding tissue and which partially formed papillary structures bearing fibrous stalks and a single layer of endothelium. The capillary formation was poorly defined, and mitotic figures were frequently observed. Immunohistochemically, the endothelial cells were positive for factor VIII related antigen and vimentin, and many cells were positive for PCNA, not only in the solid proliferating area but also in the papillary proliferating area. This case represents IPEH with high proliferative activity.

A case of calcifying odontogenic cyst with numerous calcifications: immunohistochemical analysis.

The purpose of this study was to investigate a case of calcifying odontogenic cyst (COC) in which numerous calcifications were observed not only in the lining epithelium, but also in the cyst wall, using cytokeratins 13 (CK13), 19 (CK19), and core binding factor a-1 (cbfa-1) as primary antibodies. Cells of Malassez's epithelial rest were stained as controls. Cells of the epithelial nests in the cyst wall were reactive for CK13, but their CK19 staining was similar to that observed in the lining epithelial cells. Calcifying nodules were reactive only for CK13. Cells of Malassez's epithelial rest were reactive for CK19 but not for CK13. Cbfa-1 positive reactivity was observed only in nuclei of spindle cells in the periodontal ligament. CK13 was positive superficial to the prickle cells. CK19 was positive in the basal cells of the oral mucosa. In the lining epithelium of the cyst, the expressions of CK13 and CK19 were similar to their immunoreactions in the oral mucosa. These results suggest that the odontogenic epithelium differentiated into squamous epithelial cells, which began as ghost cells in the COC, and that this process depended on the dystrophic calcification of differentiated odontogenic epithelial cells, not of osteogenic cells.

The effect of Emdogain on ectopic bone formation in tubes of rat demineralized dentin matrix.

BACKGROUND: Emdogain (EMD) is made from enamel matrix proteins (EMPs) from the tooth germ of swine and propylene glycol alginate (PGA) as a matrix. The function of EMD is known to differentiate cells of the dental follicle into cementoblasts. However, little is known about the effect of EMD on mesenchymal cells in other tissue. OBJECTIVE: The purpose of this study was to investigate whether EMD has the ability to induce hard tissue when applied with or without demineralized dentin matrix. METHODS: Half of the dentin tubes prepared from rat incisors were demineralized by treatment with 0.6 N hydrochloric acid for 3 h. EMD or PGA was injected into the demineralized or non-demineralized dentin tubes, which were then transplanted into rectus abdominis muscles. Untreated dentin tubes were also transplanted as a control. Animals were killed at 7, 14 and 21 days after the implantation. RESULTS: Non-demineralized dentin tubes with or without EMD or PGA did not form any hard tissue. In the demineralized group, chondrogenesis in the PGA groups occurred earlier than in the EMD groups. The expression of vascular endothelial growth factor (VEGF) mRNA in the demineralized group with PGA at day 14 was the highest. The expression of osteopontin and osteocalcin mRNAs was higher in all groups at 21 days compared with 7 or 14 days. CONCLUSION: These results suggest that neither EMD nor PGA has the ability to induce hard tissue and that EMPs contained within EMD might aggregate on the dentin surface and inhibit the effect of the demineralized dentin matrix.