18 antibody staining were used. There was significant difference of positive reaction between non-recurred ameloblastoma and recurred ameloblastoma in PCNA and cytokeratin 17. There were no significant difference of positive reaction between non-recurred ameloblastoma and recurred ameloblastoma in p53, Ki-67 and cytokeratin 18. From the above results, it is suggested that the recurrence of ameloblastoma is related to positive reactions of PCNA and cytokeratin 17 and the progonsis of the recurrence of ameloblastoma is able to be predicted by using PCNA and cytokeratin 17.]]>
Ameloblastoma ; Jaw ; Keratin-17 ; Keratin-18 ; Keratins ; Proliferating Cell Nuclear Antigen ; Recurrence

OBJECTIVETo report two cases of glandular odontogenic cyst and examine its cytokeratin 18,19 expression.

METHODSTwo cases of glandular odontogenic cyst were reported and studied. The cytokeratin 18, 19 expression in these two cases were also investigated using immuno-histochemical staining as well as in the situ hybridization of the cyst epithelium.

RESULTSHisto-pathological examination revealed that ciliated columnar cells, squamous cells and low-columnar cells were found in the superficial layer of the lining epithelium. Several minor salivary glands, mainly composed of seromucous cells were observed near the satellite cyst. CK18 were expressed in all layers of the lining epithelium of varying intensity. CK18 was negative in lining epithelium of the daughter cyst, but CK19 was positive. CK18-mRNA was expressed in all the layers of the lining epithelium, the salivary glands and daughter cysts.

CONCLUSIONSHistological features and CK18 expression may be indicative of the possibility of salivary glandular and odontogenic differentiation.

Adolescent ; Adult ; Epithelium ; pathology ; Female ; Humans ; Keratin-18 ; metabolism ; Keratin-19 ; metabolism ; Male ; Odontogenic Cysts ; metabolism ; pathology

BACKGROUND: The monoclonal antibody M30 recognizes a neoepitope of cytokeratin 18 that's produced during the process of apoptosis, and it is reactive in formalin-fixed, paraffin-embedded tissue. The detailed nature of apoptosis in colorectal cancer is unclear, especially in regard to the MSI status and the clinicopathologic factors. The purpose of this study was to elucidate the apoptosis assessed by M30 immunoreactivity in colorectal cancer and its relationship with the MSI status and the various clinicopathologic factors of colorectal cancers. METHODS: 101 colorectal cancers were classified according to levels of MSI as 12 MSI-H, 4 MSI-L and 85 MSS. Apoptosis was quantified by immunohistochemistry with using M30 CytoDEATH anti-body. RESULTS: The apoptotic index assessed by M30 was significantly increased in the MSI-H and MSI-L colorectal cancer compared to that in the MSS colorectal cancer. Right sided colon cancer showed a significant higher apoptotic index than did the left sided colon cancer. There was also a tendency for decreased apoptosis in metastatic colorectal cancers (Duke's stage D). There was somewhat of an increase of apoptosis in colorectal cancers with mucinous carcinoma and medullary carcinoma, and also in the colorectal cancers with an increased TIL count, but this was not statistically significant. CONCLUSION: M30 immunoreactivity is a valuable method to detect apoptosis in formalin-fixed, paraffin-embedded tissue and it might explain that MSI-H colorectal cancer shows better clinical behavior than MSS colorectal cancer in regard to the increased apoptosis.
Adenocarcinoma, Mucinous ; Apoptosis* ; Carcinoma, Medullary ; Colonic Neoplasms ; Colorectal Neoplasms* ; Immunohistochemistry ; Keratin-18 ; Microsatellite Instability

OBJECTIVETo examine cytokeratin 18(CK18) and it's gene in jaw odontogenic keratocyst (OKC) epithelial lining.

METHODSThe epithelial linings of 32 cases were subject to monoclonal antibody immunohistochemical staining for CK18, CK8 and CK19. RT-PCR and in situ hybridization for CK18 mRNA were conducted in 12 of 32 cases in keratocyst epithelial cell linings.

RESULTSIn 17 cases, CK18 were observed in keratinized surface layers, though weakly positive. In 27 cases, CK18 were positive in the granular cell layers. CK18 were also positive in the spinous cell layers in 14 cases. In all cases, CK18 was negative in basal cell layers. By RT-PCR, 4 cases expressed CK18 strongly, 8 cases weakly. By in situ hybridization, 8 cases expressed CK18 mRNA positively in both spinous and granular cell layers, and 4 cases positively in basal and keratinized cell layers. CK8 were expressed in basal cell layers of keratocyst epithelial linings. In 23 cases, CK19 were expressed in surface cell layers of keratocyst epithelial linings.

CONCLUSIONThe expression of CK18 in keratocyst epithelial linings transfers from basal cell layer to spinous layer. The expression of CK18 immunohistochemical staining and CK18 mRNA in situ hybridization are different, which shows CK18 might be related to proliferation of OKC epithelial linings. That suggests the existence of regulation of CK18 and CK18 mRNA expression.

Epithelial Cells ; Humans ; In Situ Hybridization ; Keratin-18 ; Keratins ; Odontogenic Cysts ; RNA, Messenger

Objective: To explore a simple and feasible method for the isolation and purification of hepatocytes, hepatic stellate cells (HSC), and lymphocytes from mice. Methods: The cell suspension was obtained from male C57bl/6 mice by hepatic perfusion through the portal vein digestion method and then isolated and purified by discontinuous Percoll gradient centrifugation. Trypan blue exclusion was used to determine cell viability. Glycogen staining, cytokeratin 18, and transmission electron microscopy were used to identify hepatic cells. Immunofluorescence was used to detect α-smooth muscle actin combined with desmin in HSCs. Flow cytometry was used to analyze lymphocyte subsets in the liver. Results: After isolation and purification, about 2.7×10(7) hepatocytes, 5.7×10(5) HSCS, and 4.6×106 hepatic mononuclear cells were obtained from the liver of mice with a body weight of about 22g. The cell survival rate in each group was > 95%. Hepatocytes were apparent in glycogen deposited purple-red granules and cytokeratin 18. Electron microscopy showed that there were abundant organelles in hepatocytes and tight junctions between cells. HSC had expressed α-smooth muscle actin and desmin. Flow cytometry showed hepatic mononuclear cells, including lymphocyte subsets such as CD4, CD8, NKs, and NKTs. Conclusion: The hepatic perfusion through the portal vein digestion method can isolate multiple primary cells from the liver of mice at once and has the features of simplicity and efficiency.
Male ; Mice ; Animals ; Keratin-18 ; Actins ; Desmin ; Liver ; Hepatocytes ; Hepatic Stellate Cells

OBJECTIVE: To identify the chaperone of polypyrimidine tractor-binding protein-associated splicing factor (PSF) in myeloid leukemia cells, and to explore the mechanism and redistributive pattern to cell surface of PSF in chemo-sensitive HL60 cells and resistant HL60/DOX cells. METHODS: The eukaryotic expression vector of PSF was transfected with liposomes transiently, then flow cytometry was used to detect the expression level of PSF on the cell surface 24 h, 48 h and 72 h after vector transfections. We constructed a chimeric expression vector, streptavidin binding peptide (SBP)-PSF, meanwhile this vector was transfected and made SBP-PSF fusion protein overexpress. In addition, we used streptavidin magnetic beads to precipitate the cellular chaperonin of PSF and then identified its chaperonin by mass spectrometry (MS). Lentiviral vectors containing cytokeratin18 (K18) interference sequences were transfected into 293T cells to prepare lentivirus. HL60 and HL60/DOX cells were infected with lentivirus to obtain stable interfering K18 cell lines. Next, flow cytometry was used to test the membrane relocation level of PSF. Together, these methods confirmed the similar or different mechanisms of the PSF redistributing to membrane synergistically mediated by K18 in HL60 and HL60/DOX cells. RESULTS: The expression of membrane relocated PSF was detected every day for three days (at the end of 24 h, 48 h and 72 h) after transient overexpression. The expressing rate of PSF on the cell surface was 22.4%±3.5%, 37.9%±6.0%, 58.3%±8.8%, respectively in sensitive HL60 cells, while that was 4.7%±0.5%, 3.9%±0.6%, 2.9%±0.6% , respectively in resistant HL60/DOX cells. The difference of expressing rate on each day was significant, P<0.01. We identified K18 detected by co-immunoprecipitation and mass spectrum assay which was the cellular chaperone of PSF. We found that K18 knockdown decreased the PSF expression level which redistributed on cell surface from 48.9%±5.4% to 6.2%±1.0% in sensitive HL60 cells, and from 9.11%±1.2% to 2.21%±0.51% in resistant HL60/DOX cells, respectively. CONCLUSION K18 is the intracellular chaperonin of PSF. The interaction of PSF and K18 mediates its redistribution to cell membrane in sensitive cells. While in resistant cells, PSF failed to relocate at the cell surface and accumulated in cells, which mediated resistance to chemotherapeutics.
Cell Membrane ; Doxorubicin ; Drug Resistance, Multiple ; Humans ; Keratin-18/metabolism* ; Leukemia, Myeloid

BACKGROUND: Liver biopsy for the diagnosis of non-alcoholic steatohepatitis (NASH) is limited by its inherent invasiveness and possible sampling errors. Some studies have shown that cytokeratin-18 (CK-18) concentrations may be useful in diagnosing NASH, but results across studies have been inconsistent. We aimed to identify the utility of CK-18 M30 concentrations as an alternative to liver biopsy for non-invasive identification of NASH. METHODS: Individual data were collected from 14 registry centers on patients with biopsy-proven non-alcoholic fatty liver disease (NAFLD), and in all patients, circulating CK-18 M30 levels were measured. Individuals with a NAFLD activity score (NAS) ≥5 with a score of ≥1 for each of steatosis, ballooning, and lobular inflammation were diagnosed as having definite NASH; individuals with a NAS ≤2 and no fibrosis were diagnosed as having non-alcoholic fatty liver (NAFL). RESULTS: A total of 2571 participants were screened, and 1008 (153 with NAFL and 855 with NASH) were finally enrolled. Median CK-18 M30 levels were higher in patients with NASH than in those with NAFL (mean difference 177 U/L; standardized mean difference [SMD]: 0.87 [0.69-1.04]). There was an interaction between CK-18 M30 levels and serum alanine aminotransferase, body mass index (BMI), and hypertension ( P  < 0.001, P  = 0.026 and P  = 0.049, respectively). CK-18 M30 levels were positively associated with histological NAS in most centers. The area under the receiver operating characteristics (AUROC) for NASH was 0.750 (95% confidence intervals: 0.714-0.787), and CK-18 M30 at Youden's index maximum was 275.7 U/L. Both sensitivity (55% [52%-59%]) and positive predictive value (59%) were not ideal. CONCLUSION This large multicenter registry study shows that CK-18 M30 measurement in isolation is of limited value for non-invasively diagnosing NASH.
Humans ; Non-alcoholic Fatty Liver Disease/diagnosis* ; Keratin-18 ; Biomarkers ; Biopsy ; Hepatocytes/pathology* ; Apoptosis ; Liver/pathology*

Adenoma, Sweat Gland ; metabolism ; Diagnosis, Differential ; Humans ; Keratin-14 ; metabolism ; Keratin-17 ; metabolism ; Keratin-18 ; metabolism ; Keratin-7 ; metabolism ; Keratins ; metabolism ; Papilloma ; metabolism ; Sebaceous Gland Neoplasms ; metabolism ; Skin Neoplasms ; metabolism ; Sweat Gland Neoplasms ; metabolism

Aged ; Aged, 80 and over ; Carcinoma, Hepatocellular ; diagnosis ; pathology ; surgery ; Humans ; Keratin-18 ; analysis ; Keratin-19 ; analysis ; Male ; Pancreatic Neoplasms ; diagnosis ; pathology ; surgery ; alpha-Fetoproteins ; analysis

OBJECTIVETo study the mechanism of liver fibrogenesis and to find new non-invasive biomarkers.

METHODIn this study, we used subcellular proteomic technology to study the plasma membrane proteins related to immune or alcohol induced liver fibrosis. Rat liver fibrosis models were induced by pig serum or alcohol injection. The liver fibrogenesis were detected by James's staining in the rat models after 2, 4, 6 and 8 weeks of treatment. The liver plasma membrane (PM) of the 2- and 8-week treatment model rats were enriched by two-step sucrose density gradient centrifugation. The purity of PM was verified by western blotting, and the plasma membrane proteins were extracted and analyzed by 2 DE. The differentially expressed proteins were identified by LC-MS/MS. Cellular location and function of these identified differential protein were classified.

RESULTSImmune or alcohol induced liver fibrosis rat models were successfully established. Liver plasma membrane was significantly enriched after sucrose density ultracentrifugation treatment. 87 differential protein spots were find out by 2DE combined with LC-MS/MS from the liver plasma membrane proteins of the 2- and 8-week treatment rat models, which corresponded to 30 non-redundant proteins including annexin A2, keratin 8 and keratin 18.

CONCLUSIONSA list of differentially expressed proteins relate to liver fibrosis were successfully identified. Differential proteins such as annexin A2, keratin 8 and keratin 18 could be new biomarkers for liver fibrosis diagnosis.

Alcohols ; adverse effects ; Animals ; Female ; Keratin-18 ; metabolism ; Keratin-8 ; metabolism ; Liver ; metabolism ; pathology ; Liver Cirrhosis ; chemically induced ; immunology ; metabolism ; pathology ; Male ; Proteome ; metabolism ; Rats ; Rats, Sprague-Dawley