1.Morphology, immunohistochemistry and hTERC gene in-situ hybridization in Barrett's esophagus.
Jin WANG ; Lu-ping WANG ; Sheng XU ; Guang-zhi YANG
Chinese Journal of Pathology 2013;42(1):4-9
OBJECTIVETo study the clinicopathologic features and differential diagnosis of proximal gastric mucosa and mucosa of Barrett's esophagus (BE) in biopsy specimens.
METHODThirty-eight cases of Barrett's esophagus (diagnosed using WHO criteria) and 44 cases of proximal gastric mucosa were studied by immunohistochemistry (for CK7, CK20, CK4, CK8, S-100 protein, MUC6, COX2 and bcl-2) and fluorescence in-situ hybridization (FISH) (for hTERC gene). The pathologic features were analyzed.
RESULTSThe differences in expression of CK7, CK20, MUC6, COX2 and bcl-2 between BE and proximal gastric mucosa with intestinal metaplasia were not statistically significant (P > 0.05). There was however a statistically significant difference in expression of S-100 protein (P < 0.05). The expression of CK7/CK4 and CK7/CK8 in BE showed positive correlation (P < 0.05). However, such correlation was not demonstrated in proximal gastric mucosa (P > 0.05). The results of hTERC gene expression by FISH showed a statistically significant difference between the two groups: 57.9% (22/38) in BE and 13.6% (6/44) in proximal gastric mucosa (P < 0.05).
CONCLUSIONSThe significance of CK7 and CK20 expression is uncertain in the differential diagnosis between BE and proximal gastric mucosa. On the other hand, positivity for CK7/CK4/CK8 may support the diagnosis of BE and play a role in distinguishing between the two groups. S-100 protein expression and detection of hTERC gene amplification also contribute to the diagnosis of BE.
Barrett Esophagus ; genetics ; metabolism ; pathology ; Gastric Mucosa ; metabolism ; pathology ; Gene Amplification ; Humans ; Immunohistochemistry ; In Situ Hybridization, Fluorescence ; Keratin-20 ; metabolism ; Keratin-4 ; metabolism ; Keratin-7 ; metabolism ; Keratin-8 ; metabolism ; Metaplasia ; genetics ; metabolism ; pathology ; RNA ; genetics ; Retrospective Studies ; S100 Proteins ; metabolism ; Telomerase ; genetics
2.Short-term Efficacy of Topical Immunosuppressive Agents on the Survival of Cultivated Allo-Conjunctival Equivalents.
Young Joo SHIN ; Mee Kum KIM ; Joo Youn OH ; Won Ryang WEE ; Jin Hak LEE ; Jung Hwa KO ; Hyun Ju LEE ; Jae Lim LEE ; Byung Moo MIN ; Young Suk SOHN
Korean Journal of Ophthalmology 2008;22(2):123-129
PURPOSE: To investigate the short-term efficacy of topical immunosuppressive agents on the survival of cultivated allo-conjunctival equivalents. METHODS: Twenty-five eyes of New Zealand white rabbits were included. Temporal conjunctivae were trephined to a diameter of 7.5 mm, and then cultured allo-conjunctival epithelial cells on amniotic membrane were transplanted onto them. Various immunosuppressants including steroid, cyclosporine, and rapamycin were applied topically four times a day for a week. Epithelial defects and graft edema were graded daily. Numbers of inflammatory cells were measured in H&E. PKH26 and cytokeratin 4 and 7 were immunostained. RESULTS: Earlier epithelialization was observed in 1% steroid-treated eyes and defects persisted significantly in 0.5% CsA applied eyes. In histology, PKH26 positive cells considered as donor cells were only found in 1% steroid or 0.01% rapamycin applied eyes. 1% steroid- or 0.01% rapamycin-applied eyes both showed positive staining for keratin-4 and -7. Inflammatory cells were less found in 1% steroid or 0.01% rapamycin treated eyes. CONCLUSIONS: Topical steroid or rapamycin can help to suppress acute inflammation and enhance the acute survival of transplanted conjunctival cells.
Administration, Topical
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Animals
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Cell Count
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*Cell Transplantation
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Cells, Cultured
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Conjunctiva/*cytology
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Cyclosporine/pharmacology
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Epithelial Cells/metabolism/*transplantation
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Female
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Fluorescent Antibody Technique, Indirect
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Graft Survival/*drug effects
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Immunosuppressive Agents/*pharmacology
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Keratin-4/metabolism
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Keratin-7/metabolism
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Male
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Organic Chemicals/metabolism
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Prednisone/pharmacology
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Rabbits
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Sirolimus/pharmacology
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Transplantation, Homologous
3.Diagnostic value of cytokeratin 19 fragment in nasopharyngeal carcinoma.
Dan-sheng LEI ; Jing YU ; Xian-li TONG ; Ming-wei WANG ; Kun WANG ; Hu CHEN
Chinese Journal of Pathology 2012;41(7):461-465
OBJECTIVETo explore the clinical significance of cytokeratin 19 fragments test in the diagnosis of nasopharyngeal carcinoma.
METHODSThe study included 102 cases of nasopharyngeal carcinoma, 90 cases of nasal polyp/nasopharyngitis, and 150 healthy individuals. RT-PCR was used to detect CK19 mRNA expression and Western blot to detect CK19 fragment protein expression in tissues of nasopharyngeal carcinoma. Expression of CK19-2G2 was examined by immunohistochemistry. Chemiluminescence analysis was used to detect the serum levels of CK19-2G2, and ELISA to detect that of EB-VCA IgA.
RESULTSAmong 102 cases of nasophryngeal carcinoma, 64 showed CK19 mRNA expression by RT-PCR, 60 showed CK19 protein fragments in tumor tissues by Western blot, and 66 showed expression of CK19-2G2 by immunohistochemistry in nasopharyngeal carcinoma, including strong positivity in 20 cases, moderate in 34 cases and weak in 12 cases. The sensitivity and specificity of CK19-2G2 in the diagnosis of nasopharyngeal carcinoma were 49.0% and 89.2%, and those of EB-VCA IgA were 52.9% and 85.4%, respectively. The combined detection of CK19-2G2 and EB-VCA IgA increased the sensitivity to 73.5% while the specificity remained at 80.0%.
CONCLUSIONSHigh levels of CK19-2G2 fragment expressed in tissue and serum are present in patients with nasopharyngeal carcinoma. The serum level of CK19-2G2 is helpful in the diagnosis of nasopharyngeal carcinoma. Furthermore, the combination of serum CK19-2G2 and EB-VCA IgA improves the detection sensitivity.
Adult ; Aged ; Antigens, Viral ; blood ; Blotting, Western ; Capsid Proteins ; blood ; Carcinoma, Squamous Cell ; diagnosis ; metabolism ; pathology ; Herpesvirus 4, Human ; immunology ; Humans ; Immunoglobulin A ; blood ; Immunohistochemistry ; Keratin-19 ; blood ; metabolism ; Middle Aged ; Nasopharyngeal Neoplasms ; diagnosis ; metabolism ; pathology ; Neoplasm Staging ; Peptide Fragments ; blood ; metabolism ; RNA, Messenger ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Sensitivity and Specificity
4.Hepatocyte growth factor and fibroblast growth factor-4-induced differentiation of human bone marrow mesenchymal stem cells into hepatocyte-like cells in vitro.
Jin-min XIE ; Jian-feng CHEN ; Yi GAO ; Kun-hou YAO
Journal of Southern Medical University 2006;26(10):1439-1442
OBJECTIVETo induce the differentiation of human bone marrow mesenchymal stem cells (HMSCs) into hepatocyte-like cells with hepatocyte growth factor (HGF) and fibroblast growth factor-4 (FGF-4) in vitro.
METHODSHMSCs were induced to differentiate into hepatocyte-like cells by HGF (group B), FGF-4 (group C) and HGF+FGF-4 (group D) in vitro. Undifferentiated HMSCs and L-02 cells were used as the negative (group A) and positive (group E) controls, respectively. The changes of cell morphology were observed microscopically. The expressions of hepatic markers, alpha fetoprotein (AFP) and CK-18, were detected by immunocytochemical staining at different times after induction, and the differentiation ratios of the various groups of HMSCs were calculated on the basis of image analysis. The expressions of AFP and ALB were detected by immunofluorescence assay in each group at different times after induction, and the expressions of AFP and ALB mRNA by RT-PCR.
RESULTSHMSCs gradually transformed into spindle-shaped, round, polygonal or irregular cells after induction. Immunocytochemical staining revealed positive AFP and CK18 expressions in groups B, C, and D after induction as well as in group E. The positive units (PU) of AFP and CK18 in group D calculated according to image analysis were significantly higher than that of groups A, B, and C. The expressions of AFP and ALB detected by immunofluorescence were both positive after induction in all groups except group A, similar to the findings of the expressions of AFP and ALB mRNA by RT-PCR.
CONCLUSIONHMSCs can be induced to differentiate into hepatocyte-like cells by HGF, FGF-4 and their combination at certain concentrations, and the hepatocyte-like cells can express some hepatic markers such as AFP, ALB, CK18, etc. HGF+FGF-4 may achieve more effective induction of HMSC differentiation into hepatocyte-like cells, and the efficiency of HGF is greater than that of FGF-4.
Bone Marrow Cells ; cytology ; drug effects ; metabolism ; Cell Differentiation ; drug effects ; Cells, Cultured ; Fibroblast Growth Factor 4 ; pharmacology ; Hepatocyte Growth Factor ; pharmacology ; Hepatocytes ; cytology ; drug effects ; metabolism ; Humans ; Immunohistochemistry ; Keratin-18 ; biosynthesis ; Mesenchymal Stromal Cells ; cytology ; drug effects ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; alpha-Fetoproteins ; biosynthesis ; genetics