OBJECTIVE: Comparison of protein expressions by two-dimensional gel electrophoresis (2-DE) in normal cervix and squamous cell carcinoma tissues in Korean women. METHODS: Normal cervix and squamous cell carcinoma tissues were solubilized with 2-DE buffer and the first dimension of PROTEAN IEF CELL, isoelectric focusing (IEF), was performed using pH3-10 linear IPG strips of 17 cm. And then running 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and sliver stain. Scanned image was analyzed using PDQuest 2-D softwareTM. Protein spot spectrum was identified by assisted laser desorption/ionization-time of fighting (MALDI-TOF) and the protein mass spectrum identifications were performed by searching protein databases of Swiss-prot/TrEMBL, Mascot and MS-FIT. RESULTS: We found 9 up-regulation proteins (Alpha enolase, Keratin 19 type I, Keratin 20 type I, Keratin 13 type I, beta-actin, Aflatoxin B1 aldehyde reductase 1, Annexin A2, Squamous cell carcinoma antigen 2, unknown), 7 down-reguation proteins (Annexin 1, Myosin regulatory light chain 2, 14-3-3 protein epsilon, Heat shock 27 kDa protein, Hypothetical protein (DKFZP434C1715), Tumor necrosis factor receptor superfamily member 13B, Smoth muscle protein 22-alpha) and 6 up and down-regulation proteins (Tropomyosin 1, Tropomyosin 2, Tropomyosin 3, Serine (or cysteine) proteinase inhibitor, Phosphatidylinositol transfer protein alpha isoform, Src homology 3 domain-containing protein HIP-55) between normal cervix and squamous cell carcinoma cell tissues. CONCLUSION: 2-DE offers total protein expressions between normal cervix and squamous cell carcinoma cell tissues, and searching of differently expressed protein for the diagnostic markers of squamous cell carcinoma tissue.
14-3-3 Proteins ; Actins ; Aflatoxin B1 ; Aldehyde Reductase ; Annexin A2 ; Carcinoma, Squamous Cell ; Cervix Uteri ; Databases, Protein ; Down-Regulation ; Electrophoresis, Gel, Two-Dimensional* ; Electrophoresis, Polyacrylamide Gel ; Female ; Hot Temperature ; Humans ; Isoelectric Focusing ; Keratin-13 ; Keratin-19 ; Keratin-20 ; Mass Spectrometry* ; Muscle Proteins ; Myosin Light Chains ; Phospholipid Transfer Proteins ; Phosphopyruvate Hydratase ; Receptors, Tumor Necrosis Factor ; Running ; Serine ; Shock ; Sodium Dodecyl Sulfate ; Tropomyosin ; Up-Regulation ; Uterine Cervical Neoplasms*

PURPOSE: To reconstruct a cultured conjunctival equivalent that closely resembles normal conjunctival epithelium in three-dimensional culture systems. METHODS: Human conjunctival epithelial cells were cultured on dead de-epidermized dermis in the air-exposed state. After 2 weeks of culture, the sections were stained with hematoxylin and eosin. Immunohistochemical and electron microscopic studies were performed. The results were compared with those of normal conjunctiva and cultured eyelid skin equivalent. RESULTS: In the cultured conjunctival equivalent, nonkeratinizing stratified epithelium was formed similarly to normal conjunctival epithelium. Keratin 13 was expressed, but not keratin 10, in the cultured conjunctival equivalent, similarly to normal conjunctival epithelium. However, in the cultured eyelid skin equivalent, keratinizing stratified epithelium was formed. In addition, keratin 10 was expressed, but not keratin 13, contrary to those of the cultured conjunctival equivalent. In the cultured conjunctival equivalent, ultrastructurally, keratin intermediate filaments and desmosomes were found. In addition, microvilli were seen in the uppermost epithelial cells. CONCLUSIONS: These findings demonstrate that the cultured conjunctival equivalent resembles normal conjunctival epithelium morphologically, biochemically and ultrastructurally, thereby suggesting that the cultured conjunctival equivalent may have a great potential in the study of conjunctival epithelium.
Conjunctiva ; Dermis ; Desmosomes ; Eosine Yellowish-(YS) ; Epithelial Cells ; Epithelium* ; Eyelids ; Hematoxylin ; Humans ; Intermediate Filaments ; Keratin-10 ; Keratin-13 ; Microvilli ; Skin

OBJECTIVETo investigate the rules of proliferation of epithelial cells of sweat glands in deep partial-thickness burn wound and its transdifferentiation towards epidermal cells during healing process to explore its mechanisms.

METHODSTwenty-eight patients with limbs and trunk burn hospitalized in the Fourth People's Hospital of Taizhou City of Jiangsu Province and the Second Hospital of Shandong University from January 2004 to December 2007 were enrolled in the study. Tissue samples of deep partial-thickness burn wound (DPBW, n = 37), superficial partial-thickness burn wound (SPBW, n = 21), and normal skin (NS, n = 10) were harvested. Expressions of cytokeratin 10 (CK10), bcl-2, P63, CK14 and CK19 of epithelial cells in glandular secretory portion (GSP) in DPBW, SPBW and NS were detected with immunohistochemical double staining method.

RESULTSIn NS, CK19, CK14 and CK10 expressed in medium intensity in GSP epithelial cells, P63 and CK14 weakly expressed in basal myoepithelial cells, while no expression of bcl-2 or P63 was observed in all CK10 positive terminally differentiated cells. In SPBW, no change of the construction of GSP and above-mentioned proteins during healing process was observed. In DPBW, as examined on 7(th) post burn day (PBD), expression of P63 and bcl-2 in GSP epithelial cells was enhanced. In DPBW on 8 - 10 PBD, bcl-2, P63, CK19 and CK14 strongly positive solid island-like epithelial structure was formed by proliferation, migration and squamous epithelization of basal cells. Such structure, along with granulation tissue, migrated towards the superficial layer of wounds. The hyperplasia of squamous epithelium resulted in complete reepithelialization. In DPBW, bcl-2, CK14, CK19 and P63 still strongly expressed in hyper-proliferative epidermal basal and suprabasal layers on 13 - 30 day after healing.

CONCLUSIONSDuring the natural healing process of DPBW, monolayer epithelium (CK19 and CK10 positive) of GSP slowly develops into stratified squamous epithelium (bcl-2, P63, CK19, and CK14 positive), suggesting that the epithelial-epidermal transdifferentiation of GSP undergoes slow retrodifferentiation process of stem cells and transient amplifying cells, resulting in the imbalance between lagged growth of epithelium and the hyperplasia of granulation tissue, constituting one of the important mechanisms of disturbance in DPBW repair.

Adolescent ; Adult ; Burns ; metabolism ; pathology ; Cell Differentiation ; Epithelial Cells ; metabolism ; Female ; Humans ; Keratin-10 ; metabolism ; Keratin-14 ; metabolism ; Keratin-19 ; metabolism ; Male ; Membrane Proteins ; metabolism ; Stem Cells ; metabolism ; Sweat Glands ; cytology ; metabolism ; Wound Healing ; Young Adult ; bcl-2-Associated X Protein ; metabolism

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

Adenocarcinoma, Mucinous ; metabolism ; pathology ; surgery ; Aged ; Diagnosis, Differential ; Facial Neoplasms ; metabolism ; pathology ; surgery ; Gastrointestinal Neoplasms ; metabolism ; pathology ; Humans ; Keratin-19 ; metabolism ; Keratin-20 ; metabolism ; Keratin-7 ; metabolism ; Male ; Receptors, Estrogen ; metabolism ; Skin Neoplasms ; metabolism ; pathology ; surgery

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

OBJECTIVES: Cholesteatoma is a nonneoplastic destructive lesion of the temporal bone with debated pathogenesis and bone resorptive mechanism. Both molecular and cellular events chiefly master its activity. Continued research is necessary to clarify factors related to its aggressiveness. We aimed to investigate the expression of Ki-67, cytokeratin 13 (CK13) and cytokeratin 17 (CK17) in acquired nonrecurrent human cholesteatoma and correlate them with its bone destructive capacity. METHODS: A prospective quantitative immunohistochemical study was carried out using fresh acquired cholesteatoma tissues (n=19), collected during cholesteatoma surgery. Deep meatal skin tissues from the same patients were used as control (n=8). Cholesteatoma patients were divided into 2 groups and compared (invasive and noninvasive) according to a grading score for bone resorption based upon clinical, radiologic and intraoperative findings. To our knowledge, the role of CK17 in cholesteatoma aggressiveness was first investigated in this paper. RESULTS: Both Ki-67 and CK17 were significantly overexpressed in cholesteatoma than control tissues (P < 0.001 for both Ki-67 and CK17). In addition, Ki-67 and CK17 were significantly higher in the invasive group than noninvasive group of cholesteatoma (P=0.029, P=0.033, respectively). Furthermore, Ki-67 and CK17 showed a moderate positive correlation with bone erosion scores (r=0.547, P=0.015 and r=0.588, P=0.008, respectively). In terms of CK13, no significant difference was found between cholesteatoma and skin (P=0.766). CONCLUSION: Both Ki-67 and CK17 were overexpressed in cholesteatoma tissue and positively correlated with bone resorption activity. The concept that Ki-67 can be a predictor for aggressiveness of cholesteatoma was supported. In addition, this is the first study demonstrating CK17 as a favoring marker in the aggressiveness of acquired cholesteatoma.
Bone Resorption ; Cholesteatoma* ; Ear, Middle ; Humans* ; Keratin-13* ; Keratin-17* ; Keratins* ; Ki-67 Antigen ; Prospective Studies ; Skin ; Temporal Bone

The lack of sufficient oral mucosa available for intra-oral reconstruction has been dealt with by the use of skin or oral mucosa grafts harvested from donor sites but grafts requires more than one surgical procedures and could cause donor site morbidity. Many investigators have attempted to increase available soft tissue by tissue engineered skin or oral mucosa replacements for clinical applications. But, reconstructed mucosa by several methods have low physical properties such as rolling and contraction. The aims of this study were to develope an in vitro experimental model that maintains an epithelial-mesenchymal interaction by organotypic raft culture, and to characterize biologic properties of three-dimensionally cultured oral mucosa embedded with Polydioxanone mesh by histological and immunohistochemical analysis. The results were as follows; 1. Oral mucosa reconstructed by three-dimensional organotypic culture revealed similar morphologic characteristics to equvalent normal oral mucosa in the point that they show stratification and differentiation. 2. The expression of cytokeratin 10/13 and involucrin in the cultured tissue showed the same pattern with normal oral mucosa suggesting that organotypic co-culture condition is able to induce cellular differentiation. 3. After insertion of polydioxanone mesh, increased tensile strength were observed. These results suggest that three-dimensional organotypic co-culture of the oral mucosa cell lines with the dermal equvalent consisting type I collagen and fibroblasts reproduce the morphologic and immunohistochemical characteristics similar to those in vivo condition. And increased physical properties by use of polydioxanone mesh will helpful for clinical applications.
Cell Line ; Coculture Techniques ; Collagen Type I ; Fibroblasts ; Humans ; Keratins ; Models, Theoretical ; Mouth Mucosa* ; Mucous Membrane ; Polydioxanone* ; Research Personnel ; Skin ; Tensile Strength ; Tissue Donors ; Transplants

BACKGROUNDTo explore the cut-off period of subclassification and pathological features of severe hepatitis (SH).

METHODSBased on combined clinical and pathological analyses, the complete clinical and biopsy or autopsy liver tissues data from 196 cases of patients with severe hepatitis were investigated. Meanwhile, proliferative hepatocytes, cholangioepithelia and collagens were identified by a panel of monoclonal antibodies such as those against albumin, cytokeratin 18,19 and collagen I, III with immunohistochemical method.

RESULTSThe clinical and pathological analyses indicated the cut-off periods of acute, subacute and chronic SH (ASH,SSH and CSH) were (13.4+/-7.2) d, (77.4+/-69.3) d and (80.5+/-63.2) d, respectively. Among all SH cases, one case of ASH patient presented clinical manifestation and pathological changes of ASH for 21 days, however, one patient with SSH was demonstrated 12 day course by histological examination. The time of cut-off period between ASH and SSH in child cases was shorter than that in adult cases. Histologically, ASH liver tissues showed massive and/or submassive necrosis caused by one attack, with congestive sinusoid frameworks and proliferative cholangioepithelium-like hepatocytes, while SSH liver tissues presented combined fresh and old submassive or massive necrosis caused by multiple attacks, accompanied by obviously proliferative bile ducts and sinusoid framework collapse.However, the pathological changes of CSH showed ASH- or SSH-like lesions on the background of chronic liver injury.

CONCLUSIONOur data indicated that the cut-off period between ASH and SSH is in accordance with the Scheme of Viral Hepatitis Prevention and Therapy, China, published in 2000, but excluded a part of child SH cases. In our study, the authors found a few pathological features in ASH and SSH.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Collagen ; metabolism ; Female ; Hepatitis ; classification ; metabolism ; pathology ; Humans ; Keratins, Type I ; metabolism ; Liver ; pathology ; Male ; Middle Aged ; Young Adult

OBJECTIVETo study the cytokeratin 18 and 13 and their gene (CK) expression in post-operation maxillary cyst linings with metaplastic epithelium.

METHODSCK expressions were examined with immunohistochemistry in 46 post-operative maxillary cyst (POMC) which were lined with pseudostriated columnar cells only (13 cases), both kinds of columnar and squamous cells (30 cases) and squamous cells only (3 cases).

RESULTSThe expressions of CK8, CK13 and CK18 were observed in 39, 9 and all of the 43 columnar epithelial linings, respectively. Metaplastic squamous epithelia expressed more CK13 and less CK18 and CK8. Of the 33 metaplastic linings, 24 expressed CK8, 23 CK13 and 26 linings expressed CK18. The expression of CK13- and CK18-mRNA was generally correlated with the protein expression level. By in situ hybridization, CK18-mRNA expression was observed not only in 26 metaplastic linings which were positive for CK18 protein but also in five of the seven metaplastic linings which did not express CK18 protein. In addition, RT-PCR revealed an expression of CK18-mRNA in all metaplastic squamous linings although the expression level was weaker than that in the columnar epithelial linings. The CK13-mRNA was expressed in a fashion inverse to the CK18-mRNA.

CONCLUSIONSThese results indicate that CK18-mRNA is preserved through metaplasia although the protein expression decreases and metaplastic squamous cells differentiate with a decrease of CK18 and an increase of CK13 expression.

Epithelial Cells ; metabolism ; pathology ; Humans ; Jaw Cysts ; etiology ; metabolism ; pathology ; Keratin-13 ; biosynthesis ; genetics ; Keratin-18 ; biosynthesis ; genetics ; Maxillary Diseases ; etiology ; metabolism ; pathology ; Metaplasia ; metabolism ; pathology ; Postoperative Complications ; RNA, Messenger ; genetics