The tendon is a dense connective tissue that connects muscle to bone and plays an essential role in joint motion. The injured tendon heals slowly owing to its low cellularity and vascularity. This study aimed to evaluate and compare the effects of regenerative injection therapy (RIT), 20 % dextrose prolotherapy (DP), and platelet-rich plasma (PRP) injections that can promote tendon healing. Twenty-one New Zealand white rabbits were divided into the control, DP, and PRP treatment groups. The superficial digital flexor tendon (SDFT) of the right hindlimb of each rabbit was used. A round defect of 2 mm was induced. Approximately 0.2 mL of 20% dextrose and autologous PRP were injected into the proximal and distal ends of the SDFT mass. Radiographic and ultrasonographic examination and cross-sectional area (CSA) calculations were performed pre-operatively and at 2, 4, and 8 weeks. The SDFT of both limbs was transected for biomechanical and histomorphometric evaluations. The SDFT of the left limb was transected for intact control. Semi-quantitative analysis was performed to evaluate the histomorphometric properties. Additional analysis was performed using H&E, Masson's trichrome, and immunohistochemical staining. The biomechanical evaluation showed that the treatment groups had higher tensile strength compared to the defect control group, while the PRP group had higher tensile strength than the DP group. On histological examination, the treatment groups appeared to be relatively closer to the remodeling phase of the healing process than the defect control group; the characteristics of the PRP group were closer to the remodeling phase than those of the DP group. The ultrasonographic examination showed different tendencies. Increased values in the CSA were observed during the early period in the treatment groups. This study suggests that PRP and DP can promote the healing of tendon injury, and these effects were superior with PRP than that with DP.
We report an unusual case of postoperative early gastric cancer with liver metastasis mimicking pancreaticobiliary carcinoma. A 73-year-old man with early gastric cancer was transferred for endoscopic treatment. The patient underwent endoscopic submucosal dissection for the treatment of the early gastric cancer. The pathological diagnosis was adenocarcinoma with extension to the deep submucosa and some lymphatic invasion. Therefore, subsequent a subtotal gastrectomy was performed. The histological results demonstrated residual adenocarcinoma confined to the mucosa. The resection margin and lymph node metastasis were negative. Thus, he was closely monitored for recurrence every 6 months. After 2 years, he was suddenly suspected of developing liver metastasis and local recurrence. He received a liver biopsy, and the pathological result was poorly differentiated adenocarcinoma. Immunohistochemical staining suggested pancreaticobiliary carcinoma rather than metastatic adenocarcinoma from the stomach or colon, but primary focus was not found. We were sure that the recurrent stomach cancer metastasized to the liver because stomach cancer can show heterogeneous cytokeratin (CK) expression pattern with various histological features. Therefore, no single CK expression pattern has diagnostic value for distinguishing gastric carcinoma. The patient underwent chemotherapy for metastatic stomach cancer.
Purpose: The purpose of this study is to evaluate the clinical, radiological and histopathological results after microfracture surgery for degenerative arthritis of the knee. Materials and Methods: From Oct. 1997 to Dec. 1998, 48 knees in 46 patients were treated by microfracture technique. Their mean age at the time of operation was 56 years(range, 40-75 years) and mean period of follow-up study was one year(range, 7-20 months). For 24 knees in 22 patients, 'second-look' arthroscopies and biopsies were performed at 6 months following microfracture. At the last follow up clinical results were evaluated with Baumgaertner's scale. The specimens of 24 cases were stained with H-E, Safranin-O, and Masson's trichrome. Eighteen of 24 cases were stained immunohistochemically and the Western blotting test was performed on 12 cases for type II collagen. We analyzed the relationship of the Western blotting for type II collagen with clinical score, preoperative varus deformity, joint space widening in radiological result, extent of repaired articular cartilage in '2nd-look' arthroscopic findings, patient's age and weight. Results: Clinical results were excellent in 90% and good in 10%. Among the 24 knees, more than 80% of areas of chondral defect were covered with regenerated cartilage in 21 knees Histologically, the repaired tissue appears to be a hybrid of hyaline cartilage and fibrocartilage. Repaired cartilage contains variable amounts of type II collagen with immunohistochemical staining. The results of the Western blotting test were similar. The amounts of type II collagen formation had positive correlation with the extent of repaired cartilage and preoperative varus deformity. Conclusion: 'Second-look' showed that the chondral defect areas were covered with newly grown grayish white tissue. Articular cartilage repair was confirmed with histological and immunohisto-chemical study qualitatively, and the amount of type II collagen was calculated with the Western blotting test quantitatively. The exact nature and fate of repaired cartilagenous tissues need further long term follow-up study. The results of this study provide the rationale to select osteoarthritic patients indicated for microfracture surgery.
Jeon, Hye Lyun;Yi, Jung-Sun;Kim, Tae Sung;Oh, Youkyung;Lee, Hye Jeong;Lee, Minseong;Bang, Jin Seok;Ko, Kinarm;Ahn, Il Young;Ko, Kyungyuk;Kim, Joohwan;Park, Hye-Kyung;Lee, Jong Kwon;Sohn, Soo Jung
Toxicological Research
/
v.33
no.2
/
pp.107-118
/
2017
Although alternative test methods based on the 3Rs (Replacement, Reduction, Refinement) are being developed to replace animal testing in reproductive and developmental toxicology, they are still in an early stage. Consequently, we aimed to develop alternative test methods in male animals using mouse spermatogonial stem cells (mSSCs). Here, we modified the OECD TG 489 and optimized the in vitro comet assay in our previous study. This study aimed to verify the validity of in vitro tests involving mSSCs by comparing their results with those of in vivo tests using C57BL/6 mice by gavage. We selected hydroxyurea (HU), which is known to chemically induce male reproductive toxicity. The 50% inhibitory concentration ($IC_{50}$) value of HU was 0.9 mM, as determined by the MTT assay. In the in vitro comet assay, % tail DNA and Olive tail moment (OTM) after HU administration increased significantly, compared to the control. Annexin V, PI staining and TUNEL assays showed that HU caused apoptosis in mSSCs. In order to compare in vitro tests with in vivo tests, the same substances were administered to male C57BL/6 mice. Reproductive toxicity was observed at 25, 50, 100, and 200 mg/kg/day as measured by clinical measures of reduction in sperm motility and testicular weight. The comet assay, DCFH-DA assay, H&E staining, and TUNEL assay were also performed. The results of the test with C57BL/6 mice were similar to those with mSSCs for HU treatment. Finally, linear regression analysis showed a strong positive correlation between results of in vitro tests and those of in vivo. In conclusion, the present study is the first to demonstrate the effect of HU-induced DNA damage, ROS formation, and apoptosis in mSSCs. Further, the results of the current study suggest that mSSCs could be a useful model to predict male reproductive toxicity.
This study was to examine whether the in vitro friability, motility and intact acrosome of frozen-thawed bovine and human sperm can be improved by adding Pentoxifylline (PF) or Fertilization Promoting Peptide (FPP). Human semen was frozen ultra-rapidly using Test yolk-buffer (TYB) freezing medium. Additive (PF, FPP) effects in frozen-thawed bovine and human sperm were analyzed by microscopic count for sperm motility and coomassie brilliant blue staining method f3r sperm acrosome intact. The in vitro motility of frozen-thawed bovine sperm with 5 mM PF treatment group (50.0%) was significantly higher than that of control (34.0%) (P<0.05). In the frozen-thawed bovine sperm was examined, the intact acrosome rate of 50 nM FPP treatment (49.0%) was significantly higher than those of control (30.0%) and 25 nM FPP (38.0%) treatment groups (P<0.01). In human semen, when in vitro motility of sperm with PF addition prior to freezing was examined, the result of 5 mM treatment group (51.0%) was significantly higher than those of control and 2.5 mM treatment group (39.0, 40.0%) (P<0.01). In addition, 50 nM (75.5%) FPP adding in all treatment procedures for human semen freezing (before freezing, freezing and after thawing) was significant effect on maintenance of the sperm intact acrosome percentage (control: 45.0; 25 nM: 53.0; 100 nM: 68.0%) (P<0.01). Also, the intact acrosome rate of human sperm with FPP (65.0%) was significantly higher than that with PF (43.0%) (P<0.05), although sperm motility was slightly higher in PF treatment group. These results suggest that improved sperm motility and intact acrosome of frozen thawed bovine and human sperm can be obtained by addition of PF or FPP, and that the enhanced in vitro viability, motility and intact acrosome can be obtained by addition of FPP in all semen freezing procedures.
This study attempts to investigate the developmental alterations of rat cerebral cortex, and the effects of EGEE on the developmental cerebral cortex in the prenatal, postnatal and adults were examined by morphological methods and H-E staining was used for the histological changes. In the case of injection of EGEE, at 14 day of fetal phase, parietal cortex was thickest $(95{\pm}12.7\;{\mu}m)$ but, it was thinner than in the control group $(102{\pm}14.0\;{\mu}m)$ and, occipital cortex $(57{\pm}10.5\;{\mu}m)$ compared with other cortexes was the thinnest in fetal phase. In the suckling phase, each cortex grew thick quickly but, after weanning phase, the growth of the cortex slowed and the thickness of cortex was similar to that of cortex in the adult phase. At 105 day after birth, the parietal cortex was thickest $(934{\pm}21.6\;{\mu}m)$ but, decreased compared with control group $(1113{\pm}19.0\;{\mu}m)$. When EGEE was injected in intraperitoneal of rat, the number of neuroblasts per unit area was largest $(207.7{\pm}11.4/10^{-2}\;mm$ at the mantle layer of parietal cortex at 14 day of fetal phase but, decreased compared with control group $(224.2{\pm}13.8/10^{-2}\;mm$ , and the size was largest $(7.5{\pm}1.3\;{\mu}m)$ at the ependymal cell layer of occipital cortex at 3 day after birth but, decreased compared with control group $(9.0{\pm}1.2\;{\mu}m)$. Simillar to control group, the number of granular cells and pyramidal cells were largest at the II and III layer of parietal cortex, but decreased during developmental phase. The size was largest at the IV and V layer of occipital cortex but it was decreased compared with control group. When EGEE was injected in intraperitoneal of rat, the cerebral cortex from fetal phase to 3 day after birth has differentiated into the 3 layers; ependymal, mantle and marginal layer, but empty cisternaes or vacoules in the cerebral cortexes and the condensed phases of neuroblasts were appeared. From 5 day after birth, it has differentiated into the 4 layers; molecular, external granular, mixed layer of internal granular, external and internal pyramidal cells and multiformal layer but, empty cisternaes or vacoules in the granular and pyramidal cell layers were appeared and the number per unit area of neuron was decreased. In the cerebral cortex of the weaning and adult phases, division of cell layers was not clear and empty cisternae was formed in the cortex with the cells in external granular and pyramidal cell layers, was magnified or condensed around blood vessels of neurons.
Lead toxicity was evaluated in forty-five cats on a balanced diet, treated with 0(control), 10, 100(low), 1, 000, 2, 000 and 4, 000(high)ppm of lead acetate orally on a body weight basis. The objectives were to describe the gross and histopathologic changes and to demonstrate what tissue lead concentrations correlate with the known dosages of lead. In subclinical lead toxicity, greater than 80% of the absorbed lead was deposited in the bone, whereas in more acute lead toxicity, 42% of absorbed lead was deposited in the bone and 36% and 20% of absorbed lead was deposited in the kidneys and in the liver, respectively. No gross lesions were found in the nervous system. Yellow-brown colored livers appear to be associated with lead toxicity. Neuronal necrosis in the cerebrum was the most predominant histopathologic finding. Astrocytic proliferation in the cerebral gray matter was observed in 1 high dose cat. Gliosis was noted in the cerebral cortex of 6 high dose cats. Two high dose cats had demyelination in the deepest layer of the cortical gray matter of the cerebrum. Extravasation of red cells and cavitation around the vessels were found in the cerebrum of 1 high dose cat. Six high dose cats had degeneration of Purkinje cells in the cerebellum. The microscopic findings in the peripheral nerves were ambiguous. In more acute toxicity, the cats had lead inclusions in the epithelial cells of proximal tubules of the kidneys of 7 cats and hepatocytes of the liver of S cats. These inclusions could be seen wlth H&E, but were more prominent with orcein staining. Two high dose cats had granulomas and connective tissue hyperplasia between tubules of the kidneys. Periportal hepatocyte vacuolization was observed in the liver of 22 cats. Vacuolization of seminiferous tubules and a reduced number of spermatogonia(indicative of reduced spermatogenesis) were found in the testis of 5 treated cats. Cystic ovaries were observed in 3 high dose cats and poor development of oogonia was found in 2 cats. The diagnosis of lead toxicity in cats can be suspected on the basis of the histopathologic lesions described, and can be of value in contributing to a diagnosis. A reliable diagnosis of lead poisoning can be helped utilizing tissue lead analysis(post molten)
Purpose: Oral mucositis induced by radiotherapy to the head and neck area, is a common acute complication and is considered as the most severe symptom for cancer patients in the early stages of treatment. This study was proposed to establish the oral mucositis mouse model induced by a single dose of radiation for the facility of testing therapeutic candidates which can be used for the oral mucositis treatments. Materials and Methods: Fifty-five BALB/c mice were divided into four groups: control, 16 Gy, 18 Gy, and 20 Gy. Oral mucositis was induced by a single dose of radiation to the head and neck using 6 MV x-Ray from linear accelerator. After irradiation, body weight and physical abnormalities were checked daily. Tongue tissues from all groups were taken on days 1, 2, 3, 5, 7, 9, and 14, respectively and H&E staining was conducted to examine morphological changes. Results: Body weight dramatically decreased after day 5 in all irradiated mice. In the 16 Gy treatment group, body weight was recovered on day 14. The histology data showed that the thickness of the epithelial cell layer was decreased by the accumulated time after radiation treatment, up to day 9. Severe ulceration was revealed on day 9. Conclusion: A single dose of 16 Gy is sufficient dose to induce oral mucositis in Balb/C mice. Significant changes were observed in the Balb/C mice on days 7 and 9 after radiation. It is suggested that this mouse model might be a useful standard tool for studying oral mucositis induced by radiation.
The ultimate goal of periodontal therapy is the regeneration of the periodontium that have been destroyed as a result of periodontal disease. This study were done in order to determine the healing status of periodontium under Polytetrafluoroethylene and millipore fillter combined with fibrin and the effect of the guided tissue regeneration procedures were performed as follows : 1) flap operation using PTFE membrane(control group) 2) flap operation using PTFE membrane which was fixed with fibrin(experimental group 1) 3) flap operation using millipore filter which was fixed with suture(experimental group II) 4) flap operation using millipore filter which was fixed with fibrin(experimental group II) After 1, 2, 4, 8, 12 weeks, dogs were sacrificed by perfusion technique and tissue block was excised including the tooth and prepared for light microscope with H-E & Masson’s trichrome staining. The result were as follows : In control and experimental group, there is no siginificant difference on epithelial cell down growth within 1st week, but more epithelial cell downgrowth in millipore or millipore combined with fibrin group. In this experiment, there were no significant difference in new cementum and alveolar bone formation whether PTFE membrane was fixed with suture or fibrin. In control and each experimental group, bone maturation appeared in 4 weeks, bone width increased bucco-lingually in control and experimental 1 group especially. Both control group and experimental group showed mild mew cementum formation on root surface and irregular arrangement of collagen fiber at 4 weeks, that showed obvious increased cementum formation at 8 weeks, and that was observed the functional arrangement of collagen fiber between new cementum and new alveolar bone at 12 weeks.
Using the G-, C-, and NOR-banding techniques, a karyotyping for Korean Native Pig was performed. Blood samples were collected from 50 male Korean Native Pigs that had been bred at the National Livestock Research Institute and then blood cells were prepared from in vitro cultures followed by karyotyping; G-, C-, and NOR-banding patterns of metaphase chromosomes were analyzed. The karyotype of Korean Native Pig is 38, XX or XY which consists of 5 pairs of submetacentric chromosomes(Group I), 2 pairs of acrocentric chromosomes with short p-arm(Group II), 5 pairs of medium metacentric chromosomes(Group III), 6 pairs of acrocentric chromosomes(Group IV) and metacentric X and Y sex chromosomes. On GTG-banding, the Korean Native Pig exhibited a typical and identical banding pattern in each homologous chromosomes. Overall chromosomal morphology and positions of typical landmarks of the Korean Native Pig were virtually identical to those of Committee for the Standardized Karyotype of the Domestic Pig(CSKDP). However, numbers of G-bands of the Korean Native Pig chromosomes were more than those of CSKDP. In chromosomes 1, 3, 5, 6, 7, 8, 13, 14, 15, 16, 17, 18 and X, the Korean Native Pig exhibited more separated bands as compared with CSKDP. In C-banding patterns, although the quantity of heterochromatin was variable in each chromosome, most of the Korean Native Pig chromosomes had heterochromatic C-bands on centromeres. However, the heterochromatic C-band was constantly observed on the whole Y chromosome. In AgNOR staining, the NORs were located at centromeres on the chromosomes 8 and 10. The number of NORs per metaphase ranged from 2 to 4 giving a mean value of 2.13. The number of NORs were distributed on all chromosome pair 10 but not on chromosome 8. The sizes of NORs were also differed between homologous chromosomes 8. Numbers of NORs of Korean Native Pig were significantly higher than those of Yorkshire. The pattern of pig NORs was polymorphic in breeds, individuals and cells, especially on chromosome 8.
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