BACKGROUND: Neointimal ingrowth rather than stent recoil has thought to be important for coronary arterial in-stent restenosis. Intuitively cell migration and extracellular matrix (ECM) formation seems to be important in the pathogenesis of stent restenosis. Therefore, with specific aim of identifying molecules implicated in cell migration and extracellular matrix formation, histopathologic analysis on atherectomized coronary arterial in-stent restenotic tissue was performed. METHODS: In the present study we analyzed 29 atherectomized coronary arterial in-stent restenotic tissue specimens (LAD 14, LCX 5, RCA 10) retrieved (5.7+/-5.4 months after stent deployment) from 25 patients (age 59+/-13, M/F:18/70) in whom restenosis complicated previous revascularization with Palmaz-Schatz stent. Histopathologic analysis was performed after immunostaining. Antibodies against TGF- 1, hyaluronan synthase (HAS) 1, MMP1, MMP9, urokinase type plasminogen activator, PDGF receptor were used for immunostaining. RESULTS: Myxoid tissue characterized by stellate-shaped cells embedded in a loose ECM was present in 20 out of 29 specimens, and tends to decrease over time after stenting. Foci of cell poor area (48-320 cells/mm2) in a microscopic field was present in 17 out of 29 specimens, and tends to increase over time after stenting (13/16 in <4 mo vs. 4/13 in > or =4 mo, p<0.01). Various proportions of specimens show positive stained cells with respect to each antibodies: TGF 1 in 16 out of 20:HAS1 in 10 out of 13:MMP1 in 8 out of 16:MMP9 in 4 out of 13:PDGF receptor in 12 out of 17 specimens. Abundant cells labled with certain antibodies (TGF 1, uPA, PDGF receptor) were frequently found in myxoid tissue. CONCLUSIONS: Myxoid tissue, frequently found in stent restenotic tissue, may be a biologically active tissue in terms of cell migration and of ECM formation. ECM accumulation tends to increase over time after stenting and may be important in pathogenesis of coronary arterial stent restenosis.
Antibodies ; Cell Movement* ; Extracellular Matrix* ; Humans* ; Hyaluronic Acid ; Receptors, Platelet-Derived Growth Factor ; Stents* ; Urokinase-Type Plasminogen Activator

Biologic resurfacing of the damaged joints is an area of great interest and clinical promise because of the limited potential ofdamaged articular cartilage healing. Several methods such as spongiolization. joint dehridement and ahrasion of suhchondral hone. perichondral grafts, and osteochondral grafts have heen used to repair cartilage defects, but the results were not satisfactory. Rccently autologous chondrocyle transplantation with a pcrioslcal patch was paid an altention for its advantage , the regeneration with hyalin cartilage. But it have many disadvantages such as too expensive cost. second staged operation, and technically difficult to isolatc chondrocytes from a small volume of donor site, so we performed that a definecl cartilaee delect in the ribbit patella was treated with transplanta1ion of in virto expanded allogenic chondrocvtes and then compared with an autologous chondrocytes transplantation. Adult rabbits were used to transplant autogenously and allogenously and allogenically harvested and in vitro cultured chondrocytes into patellar chondral lesions that had been made previously 3x 3mmin size , extending down to the calcified zone. Chondrocytes were isolated in the femoral condyle of the opposite knee or othe rabbit knee. And then enzymatic digestion ( collagenase A and DNase I ) was performed for 5 hours room temperature in a spinner bottle and cells were seeded in a 25cm2 culture flask in Dulheccos modified essential medium (DMEM), supplemented with l0% fetal hovine serum (FBS). The culture medium was changed twice weekly. After 14 days of culture, the cells were isolated hy irypsinization and transplanted into previously made chondral defects with an autogenous periosteal patch taken from the medial aspect of tibia. Healing ol' the defects was assessed by gross examination, immunohistochemical stain, and light microscope with hematoxylin-eosin stain at 8, 16, and 24 weeks. Allogenic and autologous chondrocytes transplantation significantly increased the amount of newly tormed repair tissue compared to that found in control knees in which the Jesion was solely covered hy a periosteal patch. The repair tissue, however, had a tendency of incomplete bonding to adjacent cartilage. This study shows that allogenic and autologous articular chondrocytes that have heen expanded for 2 weeks in vitro can stimulate the healing phase of chondral lesion. There is no signilicant diffcrence hetween allogenic and autologous chondrocytes transplantation.
Adult ; Cartilage ; Cartilage, Articular ; Chondrocytes* ; Collagenases ; Deoxyribonuclease I ; Digestion ; Humans ; Hyalin ; Joints ; Knee ; Patella* ; Rabbits ; Regeneration ; Tibia ; Tissue Donors ; Transplants

Astrocytes are the most numerous cellular elements in the cerebrum, and they normally have a very slow turnover rate. But during regeneration after injury, they proliferate markedly resulting in astrogliosis. The extracellular matrix in the central nervous system is present in the vessel walls and in the external glia limitans as a basal lamina. The presence of an intact extracellular matrix framework is important in regeneration after injury. Understanding the properties of astrocytic proliferation will be helpful to find out new treatment for functional recovery in the central nervous system. In this study, after cryogenic injury was performed on the cerebral cortex in rats, changes in astrocytes and the extracellular matrix were observed using light microscopy, immunohistochemical stain for glial fibrillary acidic protein(GFAP), proliferating cell nuclear antigen(PCNA), fibronectin, laminin, and type IV collagen, autoradiography and electron microscopy. The results were as follows; 1) The coagulative necrosis, which followed cryogenic injury on the cerebral cortex was healed, forming a new pia mater above the lesion. 2) Some of the PCNA positive cells were astrocytes and some of the GFAP positive cells showed a positive reaction to PCNA. 3) Proliferating astrocytes labelled by autoradiography or immunohistochemical stain for PCNA reached maximal numbers 3days after the injury and they were no longer found 2 weeks after injury. 4) In autoradiography with immunohistochemical stain for GFAP, about 1% of GFAP positive astrocytes were labelled by autoradiography and in double immunohistochemical stain for PCNA and GFAP, about 8-16% of GFAP positive astrocytes were also stained by PCNA. 5) In immunohistochemical stain for fibronectin, laminin and type IV collagen, laminin and type IV collagen were present in the newly formed blood vessel walls and fibronectin showed a diffuse positive reaction within the lesion. The new pia mater was formed within 2 weeks after the injury. 6) On electron microscopic examination, basal lamina material was found in the vessel wall 1 week after the injury and at 2 weeks, a nearly complete and continuous basal lamina was formed although the thickness was uneven. According to these findings, astrocytes in the cerebral cortex of adult rats proliferate very early in the regenerative period after cryogenic injury. At 2 weeks after the injury, this regeneration ceases and the damaged basal lamina of pia mater and vessel wall were reconstituted.
Adult ; Male ; Female ; Humans ; Rats ; Animals

To obtain a useful method for the identification of mycobacteria in tissue section, we evaluated 118 cases of tuberculosis: 48 pulm onary, 14 lymph nodal and 56 synovial tuberculosis. Seventy nine of these cases underwent the culture study. Sections stained with anti-Mycobacterium bovis were compared with the results of the Zieh1-Neelsen stain and culture. The immunohistochemical stain for Mycobacterium bovis in al examined cases was not any more sensitive than the Zieh1-Neelsen stain(p>0.05). Neverthless, the immunohistochemical stain was a useful method for the localization of mycobacteria because of the striking contrast between its background and the wider dimension of a positive area. Immunoreactive areas demonstrated a few intact mycobacteria showing a positive reaction in the Zieh1-Neelsen stain. In conclusion, double staining method using the immunohischemical stain for Mycobacterium bovis and the Zieh1-Neelsen stain is an efficient technique in oder to confirm the diagnosis of tuberculosis.

It has been claimed that CNS lymphoma, a rare neoplasm accounting for only a small fraction of malignant brain tumors, occurs with increasing frequency in immunologically normal as well as immunocompromised individuals. We investigated the prognostic value of Ki-67 index, p53, and bcl-2 oncoprotein expression in relation to the clinicopathological parameters in the primary CNS lymphoma patients. The tumors were graded by Kiel classification and the Working formulation and included 33 high-grade, 4 intermediate-grade, and 5 low-grade lymphomas. The phenotype was determined in 38 cases: 30 were B cell type and 8 were T cell type. All cases displayed variable degrees of nuclear Ki-67 staining from 1.0% to 92.0% (mean 51.1%). A highly significant correlation was established between the proportion of Ki-67 positive cells and the classification into grades (p=0.0002) and phenotypes (p=0.0002). Overexpression of p53 and bcl-2 protein was found in 37.1% and 51.4% of 35 patients, respectively. And p53 expression was significantly increased in B cell type (p=0.02). On Kaplan-Meier survival curve, the phenotype, grade of tumors, and p53 and bcl-2 protein expression were not correlated with overall survival. On multivariate analyses, overall survival was independently influenced by Ki-67 index. In conclusion, it is suggested that Ki-67 proliferating index is the most important marker for predicting biologic behavior of the primary CNS lymphoma.
Brain Neoplasms ; Central Nervous System* ; Classification ; Humans ; Lymphoma* ; Lymphoma, Non-Hodgkin ; Multivariate Analysis ; Phenotype

Infantile form of histiocytosis X is commonly presented as multiorgan desseminated form such as Letterer-Siwe disease. Lymph node involvement of histiocytosis X is usually accompanied by adjacent bone or skin lesion. Solitary nodal eosinophilic granuloma without evidence of other organ involvement is very rare. A case herein report is a 11 month-old female infant presented with fever and palpable both inguinal lymph nodes. There was neither skin lesion nor hepatosplenomegaly. Laboratory evaluation was within normal range except increased alkaline phosphatase and many neutrophils in urine. Radiologic examination revealed no remarkable bone lesions. And she showed good clinical outcome without evidence of other organ involvements. On microscopic examination of inguinal lymph node it was replaced by infiltration of histiocytes mainly along the sinusoid. Some of histiocytes showed morphologic features of "histiocytosis X cell" having nuclear grooves or multilobulation. Multinulceated giant cells were frequently see. Numerous eosinphils were also infiltrated and showed multifocal microabscess formation. Immunohistochemical staining revealed that majority of histiocytes were postitive for S-100 protein but multinucleated histriocytes, phagocytic histiocytes and those around the abscess were positive for macrophage marker, suck as CD68 and alpha-1-antichymotrypsin. Interestingly some histiocytes showed positivity for both S-100 protein and macrophage marker. These results suggest that histiocytosis X is proliferative disorder of phenotypically heterogenous population of histiocytes in contrast to the theory that it is a proliferative disorder of Langerhans cells.
Infant ; Male ; Female ; Humans

Although heterotopia of pancreatic tissue is a developmental anomaly found in approximately 2% of all autopsies, pancreatic tissue within the thorax and mediastinum is uncommon. In most of these instances, the pancreatic tissue is a component of gastroenteric duplication cysts, intralobar pulmonary sequestrations or teratomas. We describe an anterior mediastinal cyst consisting entirely of pancreatic tissue. A previously healthy 27-year-old woman was admitted due to chest pain during deep inspiration. The computed tomographic scan of the thorax showed a large cyst occupying the right anterior mediastinum. The excised multilocular cystic lesion measured 12 cm in maximum diameter and contained a brown, turbid fluid. The wall was fibrotic and showed a haphazard mixture of ducts and exocrine acini without islets. The histogenesis of this lesion is unclear.
Adult ; Autopsy ; Bronchopulmonary Sequestration ; Chest Pain ; Female ; Humans ; Mediastinal Cyst ; Mediastinum* ; Pancreatic Cyst* ; Teratoma ; Thorax

We reviewed 7 cases of congenital mesoblastic nephroma (4 cases of classical mesoblastic nephroma (CMN) and 3 cases of atypical mesoblastic nephroma (AMN)) using immuno-histochemical and flow cytometric study. Results are as follows. 1) The mean tumor size was 5 (3 to 7cm)cm in CMN and 9 (7 to 10cm)cm in AMN. The AMN revealed hemorrhage and necrosis in two Of three cases. A case of AMN showed cystic change without hemorrhage and necrosis. Mitotic count ranged in 0~4/10HPF in CMN and 20-35/10HPF in AMN. 2) Immunohistochemistry for vimentin was all positive. Actin, desmin were weakly positive in CMN, but negative in AMN. The findings were consistent with myofibroblastic differentiation in CMN and AMN was considered to be the less differentiated form of CMN. 3) Flow cytometiic analysis showed diploidy in two of two CMNs and two of three AMNs. Only one AMN showed aneuploidy with DNA index of 1.41. %SG2M were 8.1 and 15.9 (mean 12.0) in CMN and 16.9, 32.9 and 19.3 (mean 22.9) in AMN, respectively. We concluded that AMN should be distinguished from CMN, clinicopathologically.

No abstract available.
Genes, myc* ; Neuroblastoma* ; Polymerase Chain Reaction*

Recently, next generation sequencing (NGS) has received attention as the ultimate genotyping method to overcome the limitations of capillary electrophoresis (CE)-based short tandem repeat (STR) analysis, such as the limited number of STR loci that can be measured simultaneously using fluorescent-labeled primers and the maximum size of STR amplicons. In this study, we analyzed 15 autosomal STR markers via the NGS method and evaluated their effectiveness in STR analysis. Using male and female standard DNA as single-sources and their 1:1 mixture, we sequentially generated sample amplicons by the multiplex polymerase chain reaction (PCR) method, constructed DNA libraries by ligation of adapters with a multiplex identifier (MID), and sequenced DNA using the Roche GS Junior Platform. Sequencing data for each sample were analyzed via alignment with pre-built reference sequences. Most STR alleles could be determined by applying a coverage threshold of 20% for the two single-sources and 10% for the 1:1 mixture. The structure of the STR in each allele was accurately determined by examining the sequences of the target STR region. The mixture ratio of the mixed sample was estimated by analyzing the coverage ratios between assigned alleles at each locus and the reference/variant ratios from the observed sequence variations. In conclusion, the experimental method used in this study allowed the successful generation of NGS data. In addition, the NGS data analysis protocol enables accurate STR allele call and repeat structure determination at each locus. Therefore, this approach using the NGS system will be helpful to interpret and analysis the STR profiles from singe-source and even mixed samples in forensic investigation.
Alleles ; DNA ; Electrophoresis, Capillary ; Female ; Gene Library ; Humans ; Ligation ; Male ; Microsatellite Repeats ; Multiplex Polymerase Chain Reaction ; Statistics as Topic