BACKGROUND AND OBJECTIVES: The aim of this study is to determine whether the hyperproliferative and hyperkeratotic characters of cholesteatoma are associated with differentiation of keratinocytes in cholesteatoma by examining the localization of marker proteins, such as involucrin, filaggrin, and cytokeratins. MATERIALS AND METHODS: Immunohistochemical study was carried out in 30 cholesteatoma tissues and 10 retroauricular skins to examine the expression of involucrin, filaggrin, cytokeratin 4, 10 and 16. The staining results were graded as negative, weakly positive (<10%), moderately positive (10-70%), and strongly positive (>70%). RESULTS: Involucrin was strongly expressed in upper spinous, granular, and corneal layer of cholesteatoma. Filaggrin was strongly expressed in granular and corneal layer of cholesteatoma. Cytokeratin 4 was expressed in basal layer of retroauricular skin, but occasionally expressed in suprabasal layer of cholesteatoma. Cytokeratin 10 was homogenously expressed in all suprabasal layer of retroauricular skin, whereas pattern of shift to surface layer was showed in cholesteatoma. Cytokeratin 16 was moderately expressed at suprabasal layer in cholesteatoma. CONCLUSIONS: It can be suggested that early differentiation of suprabasal layer may lead to hyperdifferentiation and hyperkeratosis. Different expression of cytokeratins possibly indicates the altered differentiation of cholesteatoma.
Cholesteatoma ; Intermediate Filament Proteins ; Keratin-16 ; Keratin-4 ; Keratinocytes ; Keratins ; Protein Precursors ; Proteins ; Skin

BACKGROUND AND OBJECTIVES: The aim of this study is to determine whether the hyperproliferative and hyperkeratotic characters of cholesteatoma are associated with differentiation of keratinocytes in cholesteatoma by examining the localization of marker proteins, such as involucrin, filaggrin, and cytokeratins. MATERIALS AND METHODS: Immunohistochemical study was carried out in 30 cholesteatoma tissues and 10 retroauricular skins to examine the expression of involucrin, filaggrin, cytokeratin 4, 10 and 16. The staining results were graded as negative, weakly positive (<10%), moderately positive (10-70%), and strongly positive (>70%). RESULTS: Involucrin was strongly expressed in upper spinous, granular, and corneal layer of cholesteatoma. Filaggrin was strongly expressed in granular and corneal layer of cholesteatoma. Cytokeratin 4 was expressed in basal layer of retroauricular skin, but occasionally expressed in suprabasal layer of cholesteatoma. Cytokeratin 10 was homogenously expressed in all suprabasal layer of retroauricular skin, whereas pattern of shift to surface layer was showed in cholesteatoma. Cytokeratin 16 was moderately expressed at suprabasal layer in cholesteatoma. CONCLUSIONS: It can be suggested that early differentiation of suprabasal layer may lead to hyperdifferentiation and hyperkeratosis. Different expression of cytokeratins possibly indicates the altered differentiation of cholesteatoma.
Cholesteatoma ; Intermediate Filament Proteins ; Keratin-16 ; Keratin-4 ; Keratinocytes ; Keratins ; Protein Precursors ; Proteins ; Skin

No abstract available.
Desmin* ; Intermediate Filament Proteins* ; Intermediate Filaments* ; Muscle, Skeletal* ; Muscular Diseases* ; Sciatic Nerve* ; Vimentin*

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

The expression of two intermediate filaments, nestin and vimentin, was studied in spinal cord injury (SCI) to elucidate their roles in the formation of glial scars. Rats were sacrificed 1, 4, and 7 days after induction of compression injury of the spinal cord using an aneurysm clip. The affected spinal cords were studied using antibodies against nestin and vimentin intermediate filaments. One day after spinal cord injury, some clusters of nestin-positive vessels were detected in the center of the injury, but few were seen in other cell types. Vimentin immunostaining was detected in some glial cells in the center and its level of immunoreactivity was enhanced in the ependymal cells of the central canal. On days 4 and 7 after spinal cord injury, astrocytes and some ependymal cells in the central canal were stained positively for nestin and increased expression of nestin was observed in vessels. Vimentin was detected in some macrophages and astrocytes in the lesions. Nestin was co-localized with glial fibrillary acidic protein in some glial cells in SCI. These findings imply that spinal cord cells in adult animals have embryonic capacity, and these cells are activated after injury, which in turn contributes to repair of spinal cord injury through formation of a glial scar.
Animals ; Cicatrix/pathology ; Glial Fibrillary Acidic Protein/analysis ; Immunohistochemistry ; Intermediate Filament Proteins/analysis ; Intermediate Filaments/*physiology ; *Nerve Tissue Proteins ; Neuroglia/*pathology ; Rats ; Rats, Sprague-Dawley ; Spinal Cord Injuries/*pathology ; Vimentin/analysis

PURPOSE: The aim of this experiment was to observe the phenotypic changes of intermediate filaments in skeletal muscle fibers during the degeneration and regeneration of the physical injury. MATERIALS AND METHODS: The gastrocnemius muscles of rats were physically damaged by needles and serial cryosections of the damaged muscles were prepared at 2, 4, 6, 9, 15, 21 and 35 days after injury. The cryosections were immunolabelled with desmin, vimentin and histochemically reacted with NADH-TR (nicotinamide adenine dinucleotide tetrazolium reductase). RESULTS: Myotubes, asvisualized by desmin and vimentin, appeared at 9 days after injury. The regenerative myofibers were similar to normal muscles 35 days after injury. Degeneration and regeneration occurred simultaneously and positive reactions for desmin disappeared earlier than those of vimentin. CONCLUSION: Both desmin and vimentin are strong staining tools for the evaluation of myopathy. The phenotypic patterns of intermediate filaments showed various degrees of regeneration in the early stages after physical injury.
Adenine ; Animals ; Desmin ; Intermediate Filament Proteins* ; Intermediate Filaments* ; Leg* ; Muscle Fibers, Skeletal ; Muscle, Skeletal ; Muscles* ; Muscular Diseases ; Needles ; Rats* ; Regeneration* ; Vimentin

OBJECTIVETo investigate the isolation and expansion of mesenchymal stem cells (MSCs) from human umbilical cord Wharton's jelly and their biological identities, and explore the possibility of inducing human umbilical cord-derived MSCs to differentiate into neurocyte-like cells.

METHODSThe growth and proliferative abilities of human umbilical cord-derived MSCs were observed, and their immunophenotypes were determined by flow cytometry. Salvia miltiorrhiza and beta-sulfhydryl alcohol were adopted to induce the cells to differentiate. The differentiated and undifferentiated cells were identified with immunocytochemistry. The pleiotrophin and nestin genes were measured by RT-PCR.

RESULTSA population of human umbilical cord-derived MSCs were isolated from human umbilical Wharton's jelly; they were processed to obtain a fibroblast-like population of cells and could be maintained in vitro for extended periods with stable population doubling, and they were expanded as undifferentiated cells in culture for more than 10 passages, indicating their proliferative capacity. The human umbilical cord-derived MSCs were positive for CD(29), CD(44), CD(59), CD(105), but negative or weakly expressed the markers of hematopoietic cells such as CD(14), CD(33), CD(34), CD(28), CD(45) and CD(117). The important GVHD correlation markers were negative or weakly expressed, including CD(80) (B7-1), CD(86) (B7-2), CD(40) and CD(40L). Salvia miltiorrhiza beta-sulfhydryl alcohol could induce the MSCs to express nestin, a marker of neuronal precursor stem cells at early stage of differentiation. Later, they exhibited neural phenotypes, expressing beta-tubulin III and neurofilament (NF) and glial fibrillary acidic protein (GFAP). It was confirmed by RT-PCR that the MSCs could express pleiotrophin either before or after the induction of salvia miltiorrhiza, furthermore, after the induction the expression was markedly enhanced and the nestin gene was also expressed.

CONCLUSIONThe human MSCs could be isolated from human umbilical cord Wharton's jelly, and it was easy to propagate these MSCs. The negative GVHD correlated markers might result from the fact that MSCs had no HLA barrier, which may suggest potential clinical significance. The MSCs are capable of differentiating into neurocyte-like cells and they may represent an alternative stem cell source for CNS cells transplantation.

Antigens, CD ; immunology ; Carrier Proteins ; genetics ; Cell Differentiation ; physiology ; Cells, Cultured ; Cytokines ; genetics ; Female ; Flow Cytometry ; Glial Fibrillary Acidic Protein ; metabolism ; Humans ; Immunohistochemistry ; Infant, Newborn ; Intermediate Filament Proteins ; genetics ; Male ; Mesenchymal Stromal Cells ; immunology ; metabolism ; physiology ; Nerve Tissue Proteins ; genetics ; Nestin ; Neurofilament Proteins ; metabolism ; Neurons ; metabolism ; physiology ; Pregnancy ; Reverse Transcriptase Polymerase Chain Reaction ; Tubulin ; metabolism ; Umbilical Cord ; cytology

Astrocytoma ; pathology ; Brain Neoplasms ; metabolism ; pathology ; surgery ; Diagnosis, Differential ; Ganglioneuroma ; metabolism ; pathology ; surgery ; Glial Fibrillary Acidic Protein ; metabolism ; Humans ; Intermediate Filament Proteins ; metabolism ; Male ; Nerve Tissue Proteins ; metabolism ; Nestin ; Neurons ; pathology ; Oligodendroglioma ; pathology ; Synaptophysin ; metabolism ; Vimentin ; metabolism ; Young Adult

OBJECTIVETo observe the effect of acupuncture on proliferation and differentiation of neural stem cells in brain tissues of rats with traumatic brain injuny.

METHODSThirty SD rats were randomly and equally allocated to the sham-operated, the model and the acupuncture groups. The traumatic brain injury model was established by the free drop method. For the rats in the acupuncture group, acupuncture was applied once a day for 7 days. Brain histotomy was carried out when treatments were completed. Immunohistochemical techniques were adopted to detect the cells that express nestin, neurofilament proteins (NF)-200 and glial fibrillary acidic proteins (GFAP), the markers of neural stem cells, neurons, astrocytes respectively.

RESULTSCompared to the sham-operated group, the number of nestin-positive cells and NF-200-positive cells in brain tissues was decreased significantly in the model group (P < 0.01), whereas the number of GFAP-positive cells was significantly increased P<0.01). Compared to the model group, the positive cells of nestin, NF-200, GFAP in brain tissues in the acupuncture group were increased obviously (P<0.01).

CONCLUSIONSAcupuncture can significantly increase the number of nestin-positive cells, NF-200-positive cells and GFAP-positive cells, indicating the significant increase of neural stem cells, neurons and astrocytes in number. Acupuncture can improve neuranagenesis by promoting the proliferation and differentiation of neural stem cells in brain tissues. This might be one of the mechanisms for acupuncture to treat traumatic brain injury and to promote the repair of nervous function.

Acupuncture Therapy ; Animals ; Brain ; pathology ; Brain Injuries ; pathology ; therapy ; Cell Differentiation ; Cell Proliferation ; Cerebral Cortex ; pathology ; Glial Fibrillary Acidic Protein ; metabolism ; Intermediate Filament Proteins ; metabolism ; Male ; Nerve Tissue Proteins ; metabolism ; Nestin ; Neural Stem Cells ; metabolism ; pathology ; Neurofilament Proteins ; metabolism ; Rats ; Rats, Sprague-Dawley

PURPOSE: The intermediate filament proteins, desmin and vimentin, are specific components of the cytoskeleton of striated muscle fibers and of mononuclear cells of mesenchymal origin including myoblasts, respectively. Desmin has also been found in presumptive myoblasts of mammals. The aim of this experiment was attempted to observe the phenotypic changes of intermediate filaments in skeletal muscle fibers during early stages of sciatic nerve crushing injury. MATERIALS AND METHODS: The sciatic nerves of rats were surgically crushed by hemostat and serial cryosections of soleus and extensor digitorum longus(EDL) muscles were prepared at 2, 4, 6, 8, 10, 15, 20 and 27 days after nerve injury. Serial cryosections were immunolabelled with desmin, vimentin and laminin and were histochemically reacted with NADH-TR. RESULTS: 1) Firstly, desmin positive fibers were appeared in fast-twitch type C fibers of both muscles at 6 days after nerve crushing, but were not reacted for vimentin. 2) Co-expressions of desmin and vimentin were firstly detected in fast-twitch type A fibers of EDL muscles at 8 days after nerve injury. In soleus muscles, co-expressions of desmin and vimentin were firstly seen in slow-twitch type B fibers at 10 days after nerve injury. Many atrophic fibers, that contained several central nuclei like myotubes and co-expressed desmin and vimentin, were appeared in EDL muscles at 10 days after nerve injury. Although whole regions of fibers were regenerated in EDL muscles, only peripheral regions of fibers were regenerated in soleus muscles at 15 days after nerve injury. Many atrophic fibers, co-expressed of desmin and vimentin, were appeared in EDL muscles at 20 days after nerve injury. These whole fibers represented various degrees of regenerating stages. Most of mature fibers containing several central nuclei, only expressed vimentin slightly, were seen in soleus muscles at 20 days after nerve injury. Most fibers of both muscles were matured at 27 days after nerve injury, but some fibers in EDL muscles were still in processing of degeneration and regeneration. No expressions of desmin and vimentin indicated that muscle fibers were almostly matured in soleus muscles at 27 days after nerve injury. 3) Targetoid or target fibers which informed reinnervation, were appeared firstly in soleus muscles at 20 days and were seen in both muscles at 27 days after nerve injury. All targetoid and target fibers were type B fibers. CONCLUSION: Desmin was revealed in processes of degeneration and regeneration and vimentin was appealed in regeneration process. At the same time, positive immunoreactivity of desmin and vimentin showed specific differences in degree of degeneration and regeneration according to different muscles and muscle fibers.
Animals ; Cytoskeleton ; Desmin ; Intermediate Filament Proteins* ; Intermediate Filaments* ; Laminin ; Leg* ; Mammals ; Muscle Fibers, Skeletal ; Muscle, Striated ; Muscles* ; Myoblasts ; Nerve Crush ; Nerve Fibers, Myelinated ; Nerve Fibers, Unmyelinated ; Rats* ; Regeneration* ; Sciatic Nerve* ; Vimentin