Journal of Physiology & Pathology in Korean Medicine
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v.25
no.2
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pp.257-263
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2011
This study was designed to investigate the effects of Hominis placenta pharmacopuncture treatment and electroacupuncture therapy on the functional recovery and histological change, protein expression in spinal cord injury(SCI) rats. Experimental groups were divided into the Group I(normal control rat), Group II(Non-treatment after spinal cord injury induction), Group III(Hominis placenta pharmacopuncture treatment after SCI induction), Group IV (Electroacupuncture therapy after SCI induction), Group V(Hominis placenta pharmacopuncture treatment and electroacupuncture therapy after SCI induction). After operation, rats were tested at modified Tarlov test at 1 to 3 days with divided into 4 groups, and motor behavior test(BBB locomotor rating scale, Grid walk test) was examined at 3, 7, 14, and 21 days. For the observation of damage change and size of the organized surface in muscle and spinal cord, histopathological studies were performed at 21 days by H & E stain, and BDNF & NT-3 protein expression studies were performed at 21 days. Acco rding to the results, Hominis placenta pharmacopuncture treatment and electroacupuncture therapy can play a role in facilitating recovery of locomotion following spinal cord injury. Specially, Hominis placenta pharmacopuncture treatment and electroacupuncture combimed therapy after SCI induction was most improvement in functional recovery, BDNF, and NT-3 protein synthesis.
Background: Oxidative stress is a known to be associated with in the pathogenesis of many inflammatory diseases, including periodontitis. Ursolic acid is a pentacyclic triterpenoid with has antimicrobial, antioxidative, and anticancer properties. However, the role of ursolic acid in the regulating of osteogenesis remains undetermined. This study was aimed to elucidate the crucial osteogenic effects of ursolic acid and its ability to inhibit oxidative stress by targeting the immediate early response 3 (IER3)/nuclear factor erythroid 2-related factor 2 (Nrf2) pathway. Methods: Cell proliferation was determined using water-soluble tetrazolium salt assay, cell differentiation was evaluated by alkaline phosphatase (ALP) activity, and formation of calcium nodules was detected using alizarin red S stain. Generation of reactive oxygen species (ROS) was determined using by DCFH-DA fluorescence dye in hydrogen peroxide ($H_2O_2$)-treated MG-63 cells. Expression levels of IER3, Nrf2, and heme oxygenase-1 (HO-1) were analyzed using western blot analysis. Results: Our results showed that ursolic acid up-regulated the proliferation of osteoblasts without any cytotoxic effects, and promoted ALP activity and mineralization. $H_2O_2$-induced ROS generation was found to be significantly inhibited on treatment with ursolic acid. Furthermore, in $H_2O_2$-treated cells, the expression of the early response genes: IER3, Nrf2, and Nrf2-related phase II enzyme (HO-1) was enhanced in the presence of ursolic acid. Conclusion: The key findings of the present study elucidate the protective effects of ursolic acid against oxidative stress conditions in osteoblasts via the IER3/Nrf2 pathway. Thus, ursolic acid may be developed as a preventative and therapeutic agent for mineral homeostasis and inflammatory diseases caused due to oxidative injury.
The Journal of the Korean Society for Microbiology
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v.22
no.3
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pp.197-208
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1987
Chlamydia trachomatis has now shown that this interesting intracellular parasite is a cause of nongonococcal urethritis, infantile pneumonia, pelvic inflammatory disease and epididymitis, in addition to lymphogranuloma venerum and inclusion conjunctivitis. There are several diagnostic methods for C. trachomatis, but the method using monoclonal antibody is the most sensitive and specific. The hybride cell were prepared by fusion of myeloma cell($P_3X_{63}\;Ag_8{\cdot}V_{653}$) of mouse and lymphocyte of mouse(BALB/c) that were immunized with formalin killed C. trachomatis serotype D. The cell mixtures after fusion were dispensed into 640 wells of the 96 well culture plates and continuously cultured in HAT medium for 2 weeks. The supernatants of culture media in 83(13%) wells were reacted with C. trachomatis, which were determined by enzyme-linked immunosorbent assay in 96 well microplate. The clones that secreted antibody to C. trachomatis were cloned by limiting dilution. Only six monoclones secreted antibody to C. trachomatis. The antibody titer of ascitic fluid that collected from same BALB/c mice bearing hybridoma cells was above 1:100,000. These monoclonal antibodies that were IgG reacted with elementary and reticulate bodies of all serotypes(Ba, D, E, F, G, H, J and LGV type-I) using ELISA and indirect immunofluorescence stain, but there were no cross reaction with other bacteria(coagulase negative Staphylococcus, Proteus and E. coli). We concluded these six monoclones secreted the same monoclonal antibody to C. trachomatis. The sensitivity and specificity of the monoclonal antibody compared with Microtrak(confirmatory test of C. trachomatis, Syva) was 100%, respectively.
We investigate strain transmissions of a surface bonded distributed optical fiber sensor considering strain variation according to positions. We first derive a strain transmission ratio depending on a wavelength of a strain distribution of the host structure from an analysis model. The strain transmission ratio is compared with numerical results obtained from the finite element method using ABAQUS. We find that the analytical results agree well with the numerical results. The strain transmission ratio is a function of a wavelength, i.e. the strain transmission ratio decreases (increases) as the wavelength of the host strain decreases (increases). Therefore, if an arbitrary strain distribution containing various wavelengths is given to a host structure, a distorted strain distribution will be observed in the distributed optical fiber sensor compare to that of the host structure, because each wavelength shows different strain transmission ratio. The strain transmission ratio derived in this study will be useful for accurately identifying the host strain distribution based on the signal of a distributed optical fiber sensor.
The conventional histochemical stains were used to study the properties of mucosubstances of the intestine in four teleostean species, i. e., Agramus agramus, Inimicus japonicus, Epinephelus chlorostigma, and Helicolenus dactylopterus, all belonging to the order Perciformes. The following methods were used; periodic acid Schiff (PAS), alcian blue (AB) pH at 2.5, AB pH at 1.0, AB pH at 2.5-PAS, and aldehyde fuchsin (AF) pH at 1.7-AB pH at 2.5 stain. The mucosal folds of intestines show differences in species and regions. Widely distributed in all portions studied, goblet cells situated between cylinderical epithelial cells are round or ovoid in shape. They were most densely distributed in distal intestine. In the middle and. distal intestine of Epinephelus chlorostigma and all regions of intestines of Agramus agramus, Inimicus japonicus, and Helicolenus dactylopterus, the presence of both acidic and neutral mucin was confirmed. The property of acidic mucosubstance was sialomucin. Neutral mucin was only encountered in the proximal intestine of Epinephelus chlorostigma. The amount and property of mucin showed differences in species and regions. In the distal intestine of Inimicus japonicus, the amount of acidic mucin is similar to that of neutral mucin. In all regions of intestine of Agramus agramus, proximal and middle intestine of lnimicus japonicus, middle and distal intestine of Epinephelus chlorostigma, and distal intestine of Helicolenus dactylopterus, acidic mucin occured more frequently than neutral mucin. The proximal and middle intestine of Helicolenus dactylopterus have more neutral mucin than acidic mucin.
Recently, use of natural medicine is getting more attention, and some of them are believed to be effective in the treatment of periodontitis. Among them, the seeds of safflower(Carthamus tinctrorius L.) have been proven to be effective through its use in bone diseases such as fracture and osteoporosis. During the last few years, studies using the seeds of safflower gown in Korea have been active, and it has been reported that safflower seed extract increase the proliferation and the alkaline phosphatase(ALP) activity of human periodontal ligament fibroblast(hPDLF), osteoblast, and that they promote the mineralization process. In animal studies, when safflower seed extract were administered orally new bone formation was promoted. Recently, in an effort to find out the most effective osteogenic components, among many components of the safflower seed, various safflower seed fraction extracts were obtained by multistep extraction of the safflower components using various solvents. Among these, saf-M-W fraction extracted by methanol and water was most effective in increasing osteogenic potential of osteoblasts. In this study, the effect of safflower seed fraction extract, saf-M-W, on the growth and differentiation of hPDLF and MC3T3-E1 cell was investigated. The toxicity of saf-M-W on both cells was measured using M'IT(3-(4,5dimethylthiazol-2-y1)-2,5-diphenyl tetrazolium bromide) test, and ALP activity was measured using the colorimetric assay of hPDLF. In addition, in MC3T3-El cells, the expression of ALP, bone sialoprotein(BSP) mRNA was observed using Northern blot, and the mineralized nodule formation Was observed using von Kossa stain and phase-contrast microscope. 1. In concentrations below $10{\mu}g/ml$, saf-M-W didn't show any toxicity on hPDLF and MC3T3-El cell. 2. The change in saf-M-W concentration had no effect on the ALP activity of hPDLF. 3. In MC3T-E1 cells, mRNA expressions of ALP and BSP were greater in the experimental group treated with $10{\mu}g/ml$ concentration of saf-M-W compared with the control group. 4. In MC3T3-El cells, abundance of mineralized nodules were formed in the experimental group treated with $10{\mu}g/ml$ Concentration of saf-M-W, while no mineralized nodule was formed in the control group. These results suggest that safflower seed fraction extract, saf-M-W. didn't show any toxicity on hPDLF and MC3T3-E1 cell at concentrations below $10{\mu}g/ml$ and effectively enhanced the differentiation and osteogenic potential of MC3T3-El cell.
Ferritin is known to regulate iron metabolism and maintain iron in a variety of the eukaryotic organisms. The region encoding the mature ferritin (0.47 kb, H-type) of Periserrula leucophryna was amplified using the designed primers including restriction enzyme site and termination codon and subcloned in frame to the pRBAS secretion vector containing the signal sequence, RBS, and promoter of amylase gene (E. coli-Bacillus shuttle vector), resulting in recombinant pRBAS-PLF vector. Recombinant ferritin (18 kDa) was correctly processed and secreted from Bacillus subtilis LKS strain harboring the pRBAS-PLF vector and quantitatively analyzed by SDS-PAGE and western blot, respectively. Secretion of the ferritin was optimized by culture conditions (host, medium, temperature, nitrogen source) in 3 L batch culture and 5 L jar fermenter. Finally. the ferritin was largely produced using 50 L fermenter as the following conditions; at $30^{\circ}C$, 150 rpm, 1 vvm in Bacillus subtilis LKS using PY medium. The secreted ferritin was maximally measured (approximately 177.6 ug/ml) when the cell density reached to 14.4 at $OD_{600}$ (20 h incubation). The iron binding activity was confirmed by Perls' staining in 7.5% non-denaturing gel, indicating that the multimeric ferritin (composed of 24 subunits) was formed in the culture broth after secretion. Biologically, the culture broth and powder type containing ferritin were tested for possibility as feed additive in chicken broiler. As a result, the ferritin stimulated the growth of chick broil and improved feed efficiency and production index.
In order to identify the morphological changes of tissues in mouse after irradiation. We have observed the redistribution of LDH isozymes and the morphological changes of skeletal muscle, heart, kidney, liver and testis in mouse according to variation amount with the time after the 1 Gray and 3 Gray irradiation each. As a result of H-E (hematoxylin-eosin) stain, the apoptotic bodies were more easily observed in the liver than the other tissues and the quantity of the apoptotic bodies was proportionated to radiation amount. The number of apoptotic bodies was shown the highest at 1 day in most tissues and at 7 day in testis after irradiation. TUNEL (terminal deoxyribonucleodtidyl transferase-mediated dUTP-digoxigenin nick end labeling) staining was shown the same results as H-E staining. After the irradiation, the protein content was reduced in tissues except kidney. And protein content was reduced in all tissues at the initial period of 2 hours after 3 Gy irradiation. But it increased at 7 days after irradiation. LDH (EC 1.1.1.27, lactate dehydrogenase) activity was increased mostly in tissues at the early stage after 1 Gy irradiation. The maximum activity was detected earlier stage after 1 Gy irradiation than 3 Gy irradiation. The activity of LDH $A_4$ isozyme was decreased in the skeletal muscle, heart, kidney, and testis. The activity of $B_4$ and $A_2$$B_2$ sozyme was increased in the skeletal muscle and heart, and the activity of heterotetramer isozyme was increased in kidney The activity of $A_4$ isozyme in liver was detected high level and the activity of isozyme including subunit C elevated in testis. Therefore, LDH isozyme seems to play a role of lactate oxidase in most tissues except liver after irradiation. These data support that LDH isozyme is predomintly involved in the aerobic metabolism.
Discectomy is the oldest and most commonly performed operation for the painful temporomandibular joint with internal derangement. It is the one operation for which there are longest follow-up study, with Eriksson, Silver, and Tolvanen et al reporting good results in patients about 30 years after discectomy. About 80% to 90% of patients clinically experience relief from pain and dysfunction after discectomy, and interestingly, the results may improve with time. However such as Myrhaug found headache and Poswillo supposed pain and limitation of movement as postoperative complications, even though this operation had the longest follow-up term, several aspects of discectomy remain unclear. The controversies to discectomy for TMJ with pain and dysfunction have laid emphasis only upon clinical results, and then the histomorphological study was planned to evaluate the morphologic change of TMJ after discectomy. To clarify the nature of the change through an observation on the morphologic changes of articular cartilage and subchondral bone of the condyle resulting from experimental unilateral discectomy in rabbit, the author excised the left articular discs of 12 male rabbits(control 4 rabbits), weighing about 3kg, and at 1, 3, 6, 9 weeks following surgery, harvested both(left surgical site and right nonsurgical sits) TMJ. The specimens were examined with light microscope after H-E and MT stain and the obtained results were as follows. 1. Histopathological features showed thickening of articular zone and active proliferation of fibrocartilaginous zone associated with slight proliferative zone in surgical site than control group. Also replacement of chondrocytes in calcified cartilaginous zone into bone cells was observed. 2. There were thinning of thickness of each zone of articular cartilage except calcified cartilagnous zone was observed in nonsurgical site. 3. In MT stain of condylar trabeculae, there was increased calcification in nonsurgical site than control and surgical site and the degree of calcification was similar in surgical site and control group. 4. In both site of experimental group no features of degenerative changes were observed. From the above results, it is presumed that the morphologic changes of surgical site discectomy are compensatory remodeling process characterized by an hyperplastic reaction of the articular zone and fibrocartilaginous zone filling the void created by removing the disc, and the bone changes are secondary to changes in the cartilage. Increased degree of calcification seen in condylar trabeculae of nonsurgical site results from the excessive use of condyle of that site.
Kim, Young-Hee;Jyoti, Anirban;Byun, In-Sun;Oh, Ik-Hyun;Min, Young-Ki;Yang, Hun-Mo;Lee, Byong-Taek;Song, Ho-Yeon
Journal of the Korean Ceramic Society
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v.45
no.10
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pp.618-624
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2008
Various calcium phosphate bioceramics are distinguished by their excellent biocompatibility and osteoconductivity. Especially, the exceptional biodegradability of $\beta$-TCP makes it a bone graft substitute of choice in many clinical applications. The activation of osteoclasts, differentiated from macrophage precursor cells, trigger a cell-mediated resorption mechanism that renders $\beta$-TCP biodegradable. Based on this evidence, we studied the biodegradation process of granular-type $\beta$-TCP bone graft substitute through in vitro and in vivo studies. Raw 264.7 cells treated with RANKL and M-CSF differentiated into osteoclasts with macrophage-like properties, as observed with TRAP stain. These osteoclasts were cultured with $\beta$-TCP nano powders synthesized by microwave-assisted process. We confirmed the phagocytosis of osteoclasts by observing $\beta$-TCP particles in their phagosomes via electron microscopy. No damage to the osteoclasts during phagocytosis was observed, nor did the $\beta$-TCP powders show any sign of cytotoxicity. We also observed the histological changes in subcutaneous tissues of rats implanted with granule-type $\beta$-TCP synthesized by fibrous monolithic process. The $\beta$-TCP bone graft substitute was well surrounded with fibrous tissue, and 4 months after implantation, 60% of its mass had been biodegraded. Also, histological findings via H&E stain showed a higher level of infiltration of lymphocytes as well as macrophages around the granule-type $\beta$-TCP. From the results, we have concluded that macrophages play an important role in the biodegradation process of $\beta$-TCP bone graft substitutes.
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