Prognostic Correlation Between MFG-E8 Expression Level and Colorectal Cancer.

BACKGROUND AND AIMS: Colorectal cancer (CRC) is one of the leading causes of cancer-related death all over the world. Milk fat globule-epidermal growth factor (EGF)-factor VIII (MFG-E8) was found to be highly expressed in a variety of cancers. However its role in CRC is unclear. This study investigates the expression of MFG-E8 in CRC tissues and the correlation with clinicopathological features and prognosis in CRC patients. METHODS: The expression of MFG-E8 proteins was detected by immunohistochemical staining in 90 samples of CRC. The localization of MFG-E8 in colorectal tumor was examined by immunofluorescence staining. The correlation between MFG-E8 protein expression and the clinical pathological features of CRC were evaluated by χ2 test and Fisher's exact test. The survival rates were analyzed by the Kaplan-Meier method, and the relationship between prognostic factors and patient survival was analyzed by the Cox proportional hazard models. RESULTS: Our results showed that MFG-E8 expression increased significantly in colorectal cancer compared with normal mucosa tissues (p <0.001). We further validated MFG-E8 overexpression in 6 pairs of fresh tumor and adjacent normal mucosa tissues from colorectal cancer patients by Western blot (p <0.05). Immunofluorescence staining showed that MFG-E8 accumulated in close proximity to endothelial cells in human colorectal tumor tissue. In addition, high MFG-E8 protein expression was correlated with lymph node metastasis and some pathological classifications (p <0.05). Furthermore, patients with high protein level of MFG-E8 showed shortened overall survivals (p <0.05). CONCLUSION: Our results showed that MFG-E8 could be a potential novel prognostic marker for CRC and overexpression of MFG-E8 might be involved in lymph node metastasis and angiogenesis of CRC.

[Tumors with poroid features: a clinicopathologic analysis of 45 cases].

OBJECTIVE: To investigate the clinicopathological features and the differential diagnosis of poroma and porocarcinoma. METHODS: Histopathological characteristics and clinical data of 35 cases of poroma and 10 cases of porocarcinoma were analyzed retrospectively. RESULTS: The average age of 35 patients of poroma was 48 years. The average age of 10 patients of porocarcinoma was 65 years. Both poroma and porocarcinoma occured most frequently on the scalp and face,as well as the extremities. Histologically, cases of poroma were divided into three subtypes, including classic poroma (23 cases), hidroacanthoma simplex (3 cases) and dermal duct tumor (9 cases). Residual foci of benign poroma were found in all cases of porocarcinoma, most of which were classic poroma. The malignant components showed severe dysplasia and/or stromal infiltration. CONCLUSION: The diagnosis of poroma and porocarcinoma is mainly based on the microscopic characteristics. An invasive architectural pattern and/or significant cytologic pleomorphism are the most important clues for the diagnosis of porocarcinoma. Neither focal mitotic activity nor the presence of necrosis was the diagnostic feature of porocarcinoma. Malignant transformation can occur in some cases of long existing poroma with recent, rapid tumor enlargement.

Construction of plasmids expressing Sars-CoV encoding proteins and their effects on transcription of hfgl2 prothrombinase.

SARS coronavirus (SARS-CoV) is the etiologic agent of severe acute respiratory syndrome. The aim of this study was to construct Sars-CoV membrane (M), nucleocapsid (N) and spike 2 (S2) gene eukaryotic expression plasmids, and identify their expression in vitro. Gene fragments encoding N protein, M protein and S2 protein of SARS-CoV were amplified by PCR using cDNA obtained from lung samples of SARS patients as template, and subcloned into pcDNA3.1 vector to form eukaryotic expression plasmids. SARS-CoV protein eukaryotic expression plasmids were transfected respectively into CHO cells. Immunohistochemistry was employed to detect the expression of the structural proteins of SARS-CoV in transfected cells. SARS-CoV protein eukaryotic expression plasmids were successfully constructed by identification with digestion of restriction enzymes and sequencing. M, N and S2 proteins of SARS-CoV were detected in the cytoplasm of transfected CHO cells. It was concluded that these recombinant eukaryotic expression plasmids were constructed successfully, and SARS-CoV encoding proteins could activate transcription and expression of hfgl2 gene.

Construction of Plasmids Expressing Sars-CoV Encoding Proteins and Their Effects on Transcription of Hfgl2 Prothrombinase / 华中科技大学学报（医学）（英德文版）

smids were constructed suc-cessfully, and SARS-CoV encoding proteins could activate transcription and expression of hfgl2 gene.

The nucleocapsid protein of SARS-CoV induces transcription of hfgl2 prothrombinase gene dependent on C/EBP alpha.

Fibrin deposition was universal in the lungs of SARS patients and fgl2 prothrombinase gene, a novel procoagulant, was demonstrated to express highly in a clinically relevant SARS model. To investigate whether and which structural protein of SARS-CoV induced transcription of hfgl2 prothrombinase gene, three eukaryotic expression plasmids expressing nucleocapsid protein (N), membrane protein (M) and spike protein 2 (S2) of SARS-CoV were co-transfected with hfgl2 promoter luciferase-reporter plasmids and beta-galactosidase plasmid in CHO cells, respectively. M, N and S2 protein of SARS-CoV were detected by western blotting and immunohistochemistry analysis. Further assays demonstrated that expression of hfgl2 gene was related with N protein, but not with M or S2 protein in THP-1 cells and Vero cells. N protein significantly induced functional procoagulant activity in comparison with control group. Luciferase assay showed that N protein of SARS-CoV could activate the transcription of hfgl2 promoter compared with the pcDNA3.1 empty vector. Site-directed mutagenesis and EMSA assay further demonstrated that transcription factor C/EBP alpha band with its cognate cis-element in hfgl2 promoter. The results showed that N protein of SARS-CoV induced hfgl2 gene transcription dependent on the transcription factor C/EBP alpha, which maybe contribute to the development of thrombosis in SARS.