Monocyte chemoattractant protein-1(MCP-1) is released from the macrophages and endothelial cells, regulated luteotropic and luteolytic actions of macrophages and induced luteolysis. However, the mechanisms of MCP-1 on the development and maintenance of pregnant corpora lutea are thoroughly unknown. In this experiment, TUNEL stain, ED1, ED2, and MCP-1 immunohistochemistry on the corpora lutea of pregnant rats were carried out to reveal the role of macrophages in the developing corpora lutea. In the postpartum corpora lutea, the number of macrophages was increased significantly, and the intensity of ED1 and ED2 immunoreactivity in macrophages were increased moderately, and MCP-1 immunoreactivity was also increased. In conclusion, macrophages in the postpartum corpora lutea may exert phagocytic action mainly, and the macrophages in the pregnant corpora lutea maintain the structure and function of lutein cells.
The signal transduction through tyrosine kinases play important roles in neuronal development and synaptic regulation. We carried out immunoblot analyses to study tyrosine=phosphorylated proteins in the rat cerebellar postsynaptic density (PSD), a protein-rich cytoskeletal specialization underlying beneath the postsynaptic membrane. The overall protein composition of cerebellar PSD fractions was similar to that of the forebrain’s and only a few bands were different in Coomassie stain. Immunoblot analyses with phosphtyrosine-specific antiboy (4G10) showed that there are many more tyrosine-phosphorylated proteins in the cerebellar PSD than in the forebrain PSD. Interestiingly, a major phosphotyrosine signals in cerebellar PSD fractions was associated with a 50 kD molecular size, named as PSD-50. Migration of PSD-50 coincided with that of $\alpha$CaMKII and remained in the pellet fraction after N-octylglucoside extraction. These results indicate that tyrosine phosphorylation is important in cerebellar synaptic regulation and that the PSD-50 may be same as $\alpha$CaMKIIor a new protein which is a major substrate of tyrosine kinase.
In this research the results showed that Evans blue stain causes vascular permeation at antibody injection site by the passive cutaneous anaphylaxis(PCA) screening of octpus minor so we concluded. Octopus minor causes allergy. Psedosciaena Polyactis, Raja Kenojei, Metapenaeus joyneri also showed allergenicity. Microwave and autoclaving appeared to reduced allergenicity(up to 99%) during the technological treatment processing. On the other hand, UV light didn't seem to change the protein structure of allergens affect the allergenicity. Therefore, the technological treatment processing of fish such as canning and microwave would possibly reduce the allergenicity. Among the ultrafiltration fraction of Octopus minor, Pseudosciaena Polyactis, Raja Kenojei and Metapenaeus joyneri, those fraction over 100,000 contained allergen and those under 100,000 and when screening allergenic fish went through 10,000~100,000 ultrafiltration, only the fraction of over 100,000 showed the anaphylactis activity for PCA. However whether screening fish would cause anaphylaxis in human or not is questionable. The future clinical experiment will verify this result with clinical experiment patients.
An, Byeong Kil;Ha, Young Soo;Hyun, Dong Keun;Park, Chong Oon;Kim, Joon Mee
Journal of Korean Neurosurgical Society
/
v.29
no.4
/
pp.461-470
/
2000
Objective : Many investigators have demonstrated the protective effects of hypothermia following traumatic brain injury(TBI) in both animals and humans. It has long been recognized that mild to moderate hypothermia improves neurologic outcomes as well as reduces histologic and biochemical sequelae after TBI. In this study, two immunohistochemical staining using terminal deoxynucleotidyl-transferase-mediated biotin dUTP nick end labeling(TUNEL), staining of apoptosis, and ${\beta}$-amyloid precursor protein(${\beta}$-APP), a marker of axonal injury, were done and the authors evaluated the protective effects of hypothermia on axonal and neuronal injury after TBI in rats. Material and Method : The animals were prepared for the delivery of impact-acceleration brain injury as described by Marmarou and colleagues. TBI is achieved by allowing of a weight drop of 450gm, 1 m height to fall onto a metallic disc fixed on the intact skull of the rats. Fourty Sprague-Dawley rats weighing 400 to 450g were subjected to experimental TBI induced by an impact-acceleration device. Twenty rats were subjected to hypothermia after injury, with their rectal temperatures maintained at $32^{\circ}C$ for 1 hour. After this 1-hour period of hypothermia, rewarming to normothermic levels was accomplished over 30-minute period. Following 12 hours, 24 hours, 1 week and 2 weeks later the animals were killed and semiserial sagittal sections of the brain were reacted for visualization of the apoptosis and ${\beta}$-APP. Results : The density of ${\beta}$-APP marked damaged axons within the corticospinal tract at the pontomedullary junction and apoptotic cells at the contused cerebral cortex were calculated for each animal. In comparison with the untreated controls, a significant reduction in ${\beta}$-APP marked damaged axonal density and apoptotic cells were found in all hypothermic animals(p<0.05). Conclusion : This study shows that the posttraumatic hypothermia result in substantial protection in TBI, at least in terms of the injured axons and neurons.
Purpose: The tumor suppressor gene p53 has been shown to be a factor in the carcinogenesis or progression of gastric cancer. The mutant p53 has been reported to cause a higher risk of lymph-node metastasis. Futhermore, mutation of the p53 has been linked to a poor prognosis for gastric cancer. The heat shock protein-27 (HSP27), a stress protein, has also been reported to be a poor prognostic factor in ovarian and breast cancers. However, in gastric-cancer patients, controversies exist as to its influence on the prognosis. In the present study, we used an immunohistochemical stain to observe the effects of p53 and HSP27 on the clinicopathological factors and on the prognosis for gastric-cancer patients. Materials and Methods: To evaluate the significance of p53 and HSP27 in gastric cancer patients, we analyzed 212 cases of gastric cancer (stage I.IV). Tissue samples of 212 patients were stained immunohistochemically for the mutant p53 protein and for HSP27. The correlations between protein expression and the clinicopathological factors were investigated. Results: The overall expression rates for p53 and HSP27 were $36.9\%\;and\;27.8\%$, respectively. p53 and HSP27 were correlated to each other because the HSP27 expression rate was higher in the p53-positive group (P=0.046). Statistically, the p53 and the HSP27 expression rates were significantly increased in the case of tumor invasiveness, lymphatic metastasis and vessel involvement. Therefore, they play a role in cancer progression. The 5-year survival rates of the p53-positive and the p53-negative groups were $62.8\%\;and\;60.1\%$, respectively (P=0.793) while the 5-year survival rates for the HSP27-positive and HSP27-negative groups were $54.2\%\;and\;63.1\%$, respectively (P=0.090). Conclusion: p53 and HSP27 were correlated to each other in our immunohistochemical study of gastric carcinomas and they were not independent prognostic factors in gastric- cancer patients. However, further studies are needed to determine their prognostic values for gastric-cancer patients.
The proteins of the ruminant erythrocyte membranes were analysed by polyacrylamide gel electrophoresis in sodium dodecyl sulfate, and their relations to the slow erythrocyte sedimentation rate(ESR) of the ruminants were investigated by treating the erythrocytes with proteinases such as trypsin, chymotrypsin and pronase, and glycosidases such as neuraminidase and galactosidase. Protein content in the erythrocyte membrane was $2.85{\pm}0.28$ in human, $3.60{\pm}0.41$ in Korean cattle, $3.71{\pm}0.36$ in Holstein, $4.13{\pm}0.83$ in Korean native goat and $3.94{\pm}0.56mg/ml$ in sheep, showing higher in ruminant animals than in human(p<0.01). Although the general protein profiles of the ruminant erythrocyte membranes were almost similar to that of human, all the ruminant erythrocyte membranes showed one additional protein band, called band-Q in the previous report on proteins of bovine erythrocyte membrane, which migrated electrophoretically to the mid position between band-2 and band-3 in human erythrocyte membranes. The glycoprotein profiles of ruminant erythrocyte membranes revealed by periodic acid Schiff(PAS) stain showed a marked difference from that of human. The PAS-1(glycophorin) and PAS-2(sialoglycogrotein) present in human erythrocyte membranes were almost absent from the ruminant animals. Instead, a strong PAS-positive band near the origin of the electrophorograms, which was named as PAS-B in the previous report on proteins of bovine erythrocyte membranes, was shown in the ruminant animals except sheep. In addition, the erythrocyte membranes of Korean native goat and sheep showed a moderate PAS-negative band near the tracking dye of the electrophorograms, which was named as PAS-G in this study. In the erythrocyte treated with the enzymes, the migration of each protein fracture of erythrocyte membranes in response to each enzyme was diverse according to different species or breed of ruminant animals. Among others, band-Q present in ruminants was slightly or moderately decreased by trypsin-, chymotrypsin-, and pronase- treatments of the erythrocytes, but not only in sheep. It was particularly noticeable that PAS-B, a fraction of glycoprotein, present in ruminants except sheep, was better digested by proteinases than by glycosidases, showing remarkable increase(p<0.01) of the ESR in accord with complete digestion(disappearance) of the PAS-B band by pronase, trypsin or chymotrypsin treatment of erythrocytes. In sheep, there was almost no any response to the various enzymes in general protein and glycoprotein profiles of the erythrocyte membranes except PAS-G, which was markedly decreased by pronase treatment of the erythrocytes. Nevertheless, the ESRs were accelerated in erythrocytes treated with pronase, trypsin, chymotrypsin and neuraminidase. Erythrocyte osmotic fragility was increased in erythrocytes treated with only pronase among five enzymes in all the human and ruminant animals used in this study.
Advanced glycation end products (AGEs) such as $N^{\varepsilon}$-(carboxy-methyl)lysine (CML) have been implicated in the development of diabetic nephropathy. The aim of this study was to investigate the inhibitory effects of ethanolic extract of Aster koraiensis (AKE) on AGEs formation and AGEs-collagen cross-linking in vitro and CMLs formation in streptozotocin (STZ)-induced diabetic rats. AKE significantly inhibited AGEs formation ($IC_{50}$ value of $18.74{\mu}g/mL$) and AGEs-collagen cross-linking ($IC_{50}$ value of 0.274 mg/mL) in vitro than the well-known glycation inhibitor aminoguanidine ($IC_{50}$ value of $72.12{\mu}g/mL$ and 1.99 mg/mL, respectively). AKE (100 mg/kg per day) was given to diabetic rats for 9 weeks. In STZ-induced diabetic rats, severe hyperglycemia was developed, and urinary CMLs and plasma CMLs were markedly increased. Immunohistochemical stain revealed that CMLs were accumulated within renal glomerulus in STZ-induced diabetic rats. However, AKE significantly reduced urinary CMLs and plasma CMLs in diabetic rats. CMLs accumulation was inhibited by AKE treatment in the renal glomerulus. These results suggest that AKE had an inhibitory effect of AGE accumulation in the glomeruli of diabetic rat and could be an inhibitor of AGE-induced protein cross-linking. The oral administration of AKE may significantly help to prevent the progression of diabetic nephropathy in patients with diabetes.
Ha, Yoon;Yoon, Do Heum;Yeon, Dong Su;Kim, Hyun Ok;Lee, Jin Ju;Cho, Yong Eun;Choi, Joong Uhn
Journal of Korean Neurosurgical Society
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v.30
no.8
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pp.963-969
/
2001
Objectives : Cord blood stem cells have been widely used as donor cells for bone marrow transplantation recently. These cells can give rise to a variety of hematopoietic lineages to repopulate the blood. Recent observations reveal that some bone marrow cells and bone marrow stromal cells(MSCs) can grow to become either neurons or glial cells. It is, however, unclear whether or not there exists stems cells which can differentiate into neurons in the blood during the early stages of postnatal life. Methods : Human cord blood stem cells were prepared from human placenta after full term delivery. To induce neuronal differentiation of stem cells, ${\beta}$-mercaptoethanol was treated. To confirm the neuro-glial characteristics of differentiated stem cells, immunocytochemical stain for NeuN, neurofilament, glial fibrillary acidic protein(GFAP), microtubule associated protein2(MAP2) was performed. RT-PCR was performed for detecting nestin mRNA and MAP2 mRNA. Results : We showed in this experiment that neuro-glial markers(NeuN, neurofilament, MAP2, GFAP) were expressed and axon-like cytoplasmic processes are elaborated in the cultured human cord blood stem cells prepared from new born placenta after full term delivery. Nestin mRNA was also detected in fresh cord blood monocytes. Conclusions : These results suggest that human cord blood derived stem cells may be potential sources of neurons in early postnatal life.
Ding, Ning;Lu, Yanzhu;Cui, Hanmin;Ma, Qinyu;Qiu, Dongxia;Wei, Xueting;Dou, Ce;Cao, Ning
BMB Reports
/
v.53
no.3
/
pp.154-159
/
2020
We investigated the effects of physalin A, B, D, and F on osteoclastogenesis induced by receptor activator of nuclear factor κB ligand (RANKL). The biological functions of different physalins were first predicted using an in silico bioinformatic tool (BATMAN-TCM). Afterwards, we tested cell viability and cell apoptosis rate to analyze the cytotoxicity of different physalins. We analyzed the inhibitory effects of physalins on RANKL-induced osteoclastogenesis from mouse bone-marrow macrophages (BMMs) using a tartrate-resistant acid phosphatase (TRAP) stain. We found that physalin D has the best selectivity index (SI) among all analyzed physalins. We then confirmed the inhibitory effects of physalin D on osteoclast maturation and function by immunostaining of F-actin and a pit-formation assay. On the molecular level, physalin D attenuated RANKL-evoked intracellular calcium ([Ca(2+)](i)) oscillation by inhibiting phosphorylation of phospholipase Cγ2 (PLCγ2) and thus blocked the downstream activation of Ca2+/calmodulin-dependent protein kinases (CaMK)IV and cAMP-responsive element-binding protein (CREB). An animal study showed that physalin D treatment rescues bone microarchitecture, prevents bone loss, and restores bone strength in a model of rapid bone loss induced by soluble RANKL. Taken together, these results suggest that physalin D inhibits RANKL-induced osteoclastogenesis and bone loss via suppressing the PLCγ2-CaMK-CREB pathway.
Chordoma is a slow-growing malignant neoplasm arising from the remnants of the primitive notochord. It accounts for 1 to 4% of all malignant bone tumors. It occurs exclusively along the spinal axis. Authors experienced four cases of chordoma occurred in the sacrococcygeal region. There were two male and two female patients, with a mean age of 63.5 years(range, 57~75 years). Tissue was obtained by wide excision in two patients, by incisional biopsy in one patient and by needle biopsy in the other. Adjuvant radiation therapy was performed on all the patients after their biopsy. The mean diameter of the tumors was 7.6cm(range, 5.5 to 13.0cm). Grossly, tumor was multiobulated, soft and myxoid gelatinous mass. Microscopically, the tumor showed lobulated feature divided by fibrous septa within it. There were physaliphorous cells with vacuolated bubbly cytoplasm. And small uniform, round, and non-vacuolated tumor cells were also present. On immunohistochemical stain, all the cases were immunoreactive for cytokeratin, epithelial membrane antigen(EMA) and vimentin, respectively. One of the 4 cases was positive for S-100 protein. All the cases were negative for CEA.
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