Biodegradable barrier membrane has been demonstrated to have guided bone regeneration capacity on the animal study. The purpose of this study is to evaluate the effects of cultured calvarial cell inoculated on the biodegradable barrier membrane for the regeneration of the artificial bone defect. In this experiment 35 Sprague-Dawley male rats(mean BW 150gm) were used. 30 rats were divided into 3 groups. In group I, defects were covered periosteum without membrane. In group II, defects were repaired using biodegradable barrier membrane. In group III, the defects were repaired using biodegradable barrier membrane seeded with cultured calvarial cell. Every surgical procedure were performed under the general anesthesia by using with intravenous injection of Pentobarbital sodium(30mg/Kg). After anesthesia, 5 rats were sacrificed by decapitation to obtain the calvaria for bone cell culture. Calvarial cells were cultured with Dulbecco's Modified Essential Medium contained with 10% Fetal Bovine Serum under the conventional conditions. The number of cell inoculated on the membrane were $1{\times}10^6$ Cells/ml. The membrane were inserted on the artificial bone defect after 3 days of culture. A single 3-mm diameter full-thickness artificial calvarial defect was made in each animal by using with bone trephine drill. After the every surgical intervention of animal, all of the animals were sacrificed at 1, 2, 3 weeks after surgery by using of perfusion technique. For obtaining histological section, tissues were fixed in 2.5% Glutaraldehyde (0.1M cacodylate buffer, pH 7.2) and Karnovsky's fixative solution, and decalcified with 0.1M disodium ethylene diaminetetraacetate for 3 weeks. Tissue embeding was performed in paraffin and cut parallel to the surface of calvaria. Section in 7${\mu}m$ thickness of tissue was done and stained with Hematoxylin-Eosin. All the specimens were observed under the light microscopy. The following results were obtained. 1 . During the whole period of experiment, fibrous connective tissue was revealed at 1week after surgery which meant rapid soft tissue recovery. The healing rate of defected area into new bone formation of the test group was observed more rapid tendency than other two groups. 2 . The sequence of healing rate of bone defected area was as follows ; test group, positive control, negative control group. 3 . During the experiment, an osteoclastic cell around preexisted bone was not found. New bone formation was originated from the periphery of the remaing bone wall, and gradually extended into central portion of the bone defect. 4 . The biodegradable barrier membrane was observed favorable biocompatibility during this experimental period without any other noticeable foreign body reaction. And mineralization in the newly formed osteoid tissue revealed relatively more rapid than other group since early stage of the healing process. Conclusively, the cultured bone cell inoculated onto the biodegradable barrier membrane may have an important role of regeneration of artificial bone defects of alveolar bone. This study thus demonstrates a tissue-engineering the approach to the repair of bone defects, which may have clinical applications in clinical fields of the dentistry including periodontics.
Purpose: Currently, PET/CT scan has been known to provide useful information to both preoperative and postoperative examination of cancer patients. Contracted stomach by the long fasting could cause difficulties of interpretation because of its size on reconstructed image data. To solve this problem, after the whole body PET/CT scan, patients were administrated in drinking 300 mL of water to expand stomach and performed additional scan on stomach region. Not only PET/CT scan but also CT performs this water-administration, and patients were take oral solution to make stomach expand for stomach cancer. When this scan performed, patients lay supine position. In this study, we evaluated the capacity of stomach through PET/CT scan with drinking water performed in supine and prone position so that we can distinguish exact location of cancer around pylorus and inferior wall of stomach. Furthermore, image data from supine and prone positions were analyzed the difference of volume of stomach through the change of standardized uptake values. Materials and Methods: From July 2009 to January 2010 in severance hospital, 30 patients who were diagnosed as early gastric cancer or advanced gastric cancer were chosen. All patients had PET/CT scan before the operation and have had follow-up PET/CT. The patients fast for at least 8 hours, and had an injection intravenously with $^{18}F$-FDG, 7.4 MBq (0.2 mCi/kg) per kilogram. They were rested for 60 minutes. Before the examination, all patients were administrated to drink water for 300 mL Patients had PET/CT scan with supine position around the region of stomach, whole body, and around the region of stomach with prone position after drinking another 300 mL of water respectively. Results: As a results of comparison between stomach capacity of 30 patients in supine and prone position, the study draw results that average capacity of stomach body was 460.29 $mm^2$ in supine position, and 641.39 $mm^2$ in prone position for 30 patients. The change of capacity shows 41.3% expanded in prone position. And there was no noticeable difference at maximum standardized uptake values in supine position and prone position. Conclusion: As results, stomach would have more expanded capacity in prone position than supine position. For patients who have physical disabilities to move freely, additional scan in prone position will be obstacle to perform. However, if additional scan in supine position add with the scan in prone position, it will be easier to diagnose stomach cancer. Moreover, we believe that this study will help the research for inventing support tools for patients who have physical disabilities in prone position.
Purpose : Be aware of clinical possibilities on image quality by comparison of contrast-enhanced dynamic CT and MR imaging applied of MIP technique after the experimentally induced clonorchasis infection in dogs. Materials and Method : Twenty mongrel dogs prepared in zoo-laboratory were followed up with serial CT scans and MR imaging for 13 weeks after the experimental infection in liver. Two-phase helical CT was acquired in the supine position with the following scanning parameters. After the injection of contrast material, the arterial phase was initiated using a bolus-racking method. The portal phase scan was started 15 seconds after the arterial phase scan. CT protocol was determined after single level dynamic scans. MR imaging used the CP body coil and images get a 2D image using HASTE, FLASH, TSE pulse sequence. Bile duct MR imaging were obtained in three plans. Then each image was post processed by using target MIP algorithm. Two experimentation above, as a method of evaluation, one pathologist, three radiologist and five radiological technologist were analyzed visually for evaluation of following findings, enhancement of the bile duct wall, dilatation of bile duct tip, liver parenchyma, background suppression. Results : Five dogs was died of a disease after the infection, the rest one else shows the chronic dilatation of the intrahepatic bile duct with CT and MR imaging. Contrast administration of CT shows the contrast-enhanced of the bile duct walls with live parenchyma. MR imaging calculated of CNR and CR from pulse sequence for comparative evaluation and shows the pattern of the intrahepatic bile duct, dilatation of bile duct tip using MIP technique. CNR of the clonorchiasis, HASTE was $16{\pm}0.83$, TSE $7.06{\pm}3.0$, FLASH $1.19{\pm}0.2$ and CR, HASTE was 73.3%, TSE 62.3%, FLASH 6.4%. Conclusion : CT and MR imaging is very usefulness in diagnosis of dog clonorchiasis.
This study was performed to estimate the effects of cultured bone cell inoculated on porous type hydroxyaptite for the regeneration of the artificial alveolar bone defect. In this experiment 3 beagle dogs were used, and each of them were divided into right and left mandible. Every surgical intervention were performed under the general anesthesia by using with intravenous injection of Pentobarbital sodium(30mg/Kg). To reduce the gingival bleeding during surgery, operative site was injected with Lidocaine hydrochloride(l:80,000 Epinephrine) as local anesthesia. After surgery experimental animal were feeded with soft dietl Mighty dog, Frisies Co., U.S.A.) for 1 weeks to avoid irritaion to soft tissue by food. 2 months before surgery both side of mandibular 1st premolar were extracted and bone chips from mandibular body were obtained from all animals. Bone cells were cultured from bone chips obtained from mandible with Dulbecco's Modified Essential Medium contained with 10% Fetal Bovine Serum under the conventional conditions. Porous type hydroxyapatite were immerse into the high concentrated cell suspension solution, and put 4 hours for attachin the cells on the surface of hydroxyapatite. Graft material were inserted on the artificial bone defect after 3 days of culture. Before insertion of cellinoculated graft material, scanning electronic microscopic observation were performed to confirm the attachment and spreading of cell on the hydroxyapatite surface. 3 artificial bone defects were made with bone trephine drill on the both side of mandible of the experimental animal. First defect was designed without insertion of graft material as negative control, second was filled with porous replamineform hydroxyapatite inoculated with cultured bone marrow cells as expermiental site, and third was filled with graft materials only as positive control. The size of every artificial bone defect was 3mm in diameter and 3mm in depth. After the every surgical intervention of animals, oral hygiene program were performed with 1.0% chlorhexidine digluconate. All of the animals were sacrificed at 2, 4, 6 weeks after surgery. For obtaining histological section, tissus were fixed in 10% Buffered formalin and decalcified with Planko - Rycho Solution for 72hr. Tissue embeding was performed in paraffin and cut parallel to the surface of mandibular body. Section in 8um thickness of tissue was done and stained with Hematoxylin - Eosin. All the specimens were observed under the light microscopy. The following results were obtained : 1. In the case of control site which has no graft material, less inflammatory cell infiltration and rapid new bone forming tendency were revealed compared with experimental groups. But bone surface were observed depression pattern on defect area because of soft tissue invasion into the artificial bone defect during the experimental period. 2. In the porous hydroxyapatite only group, inflammatory cell infiltration was prominet and dense connective tissue were encapsulated around grafted materials. osteoblastic activity in the early stage after surgery was low to compared with grafted with bone cells. 3. In the case of porous hydroxyapatite inoculated with bone cell, less inflammatory cell infiltration and rapid new bone formation activity was revealed than hydroxyapatite only group. Active new bone formation were observed in the early stage of control group. 4. The origin of new bone forming was revealed not from the center of defected area but from the surface of preexisting bony wall on every specimen. 5. In this experiment, osteoclastic cell was not found around grafted materials, and fibrovascular invasion into regions with no noticeable foreign body reaction. Conclusively, the cultured bone cell inoculated onto the porous hydroxyapatite may have an important role of regeneration of artificial bone defects of alveolar bone.
Four species(25%) of Viperidae(Agkistrodon brevicaudus, Agkistrodon ussuriensis, Agkistrodon saxatilis) and Cloubridae(Rhabdophis tigrenus tigrenus) were Korean poisonous snake. Copulation season of these species was from July to August. Reproduction mode of genus Agkistrodon species was ovoviviparous but Rhabdophis tigrinus tigrinus was the other pattern of oviparous. Optimal movement temperature range was from 20$\circ $C to 29$\circ $C(March~September). Wjen atmosphere temperature was below 10$\circ $C, at that time they hibernate at the ground, rock bottom, stone wall and embankment around the end of a field. The venom of these snakes consist mainly Hematoxin, Cytolysin, Neurotoxin and Cardiotoxin of poisonous liquids. These material injection to animal cause systemic syndrome such as Dizziness(25.7%), Vomitting(23.1%), Fever(22%), Visual trouble(18%), Headace(17.7%), Dyspnoea(17.6%) and bring about other local syndrome such as Discoloration(54.2%), Bleeding(20.2%), Bullae(10.7%) and Skin ulcer(!0.8%). The annual distribution was appeared to decrease 1972 after 1992 and average snakebiting patients was 25.6 per year, but practically total estimated snakebiting was 2,700 per year. The seasonal distribution was most frequent in August(25%), and mortality was 1.8%(26 per 1,430). The sex ratio was 2:1 and according to age distribution, it was most prevalent at one's fifties(19%). The most frequent place where the accident happened was the field(48.2%) and most predilection site of the body for victim were hand(47.8%) and foot(39.5%), Commonly bite snake were Agkistrodon ussuriensis(27.1%), Agkistrodon brevicaudus(22.6%) and Agkistrodon saxatilis(9.6%) but 40.7% of species could not be identified. Treatment of antivenin patient was 75.9% (1,068/1,407).
The local arrangement of sensory nerve cell bodies and nerve fibers in the brain stem, spinal ganglia and nodose ganglia were observed following injection of cholera toxin B subunit(CTB) and wheat germ agglutinin-horseradish peroxidase(WGA-HRP) into the rat intestine. The tracers were injected in the stomach(anterior and posterior portion), duodenum, jejunum, ileum, cecum, ascending colon or descending colon. After survival times of 48-96 hours, the rats were perfused and their brain, spinal and nodose ganglia were frozen sectioned ($40{\mu}m$). These sectiones were stained by CTB immunohistochemical and HRP histochemical staining methods and observed by dark and light microscopy. The results were as follows: 1. WGA-HRP labeled afferent terminal fields in the brain stem were seen in the stomach and cecum, and CTB labeled afferent terminal fields in the brain stem were seen in all parts of the intestine. 2. Afferent terminal fields innervating the intestine were heavily labeled bilaterally gelalinous part of nucleus of tractus solitarius(gelNTS), dorsomedial part of gelNTS, commissural part of NTS(comNTS), medial part of NTS(medNTS), wall of the fourth ventricle, ventral border of area postrema and comNTS in midline dorsal to the central canal. 3. WGA-HRP labeled sensory neurons were observed bilaterally within the spinal ganglia, and labeled sensory neurons innervating the stomach were observed in spinal ganglia $T_2-L_1$ and the most numerous in spinal ganglia $T_{8-9}$. 4. Labeled sensory neurons innervating the duodenum were observed in spinal ganglia $T_6-L_2$ and labeled cell number were fewer than the other parts of the intestines. 5. Labeled sensory neurons innervating the jejunum were observed in spinal ganglia $T_6-L_2$ and the most numerous area in the spinal ganglia were $T_{12}$ in left and $T_{13}$ in right. 6. Labeled sensory neurons innervating the ileum were observed in spinal ganglia $T_6-L_2$ and the most numerous area in the spinal ganglia were $T_{11}$ in left and $L_1$ in right. 7. Labeled sensory neurons innervating the cecum were observed in spinal ganglia $T_7-L_2$ and the most numerous area in the spinal ganglia were $T_{11}$ in left and $T_{11-12}$ in right. 8. Labeled sensory neurons innervating the ascending colon were observed in spinal ganglia $T_7-L_2$ in left, and $T_9-L_4$ in right. The most numerous area in the spinal ganglia were $T_9$ in left and $T_{11}$ in right. 9. Labeled sensory neurons innervating the descending colon were observed in spinal ganglia $T_9-L_2$ in left, and $T_6-L_2$ in right. The most numerous area in the spinal ganglia were $T_{13}$ in left and $L_1$ in right. 10. WGA-HRP labeled sensory neurons were observed bilaterally within the nodose ganglia, and the most numerous labeled sensory neurons innervating the abdominal organs were observed in the stomach. 11. The number of labeled sensory neurons within the nodose ganglia innervating small and large intestines were fewer than that of labeled sensory neurons innervating stomach These results indicated that area of sensory neurons innervated all parts of intestines were bilaterally gelatinous part of nucleus tractus solitarius(gelNTS), dorsomedial part of gelNTS, commissural part of NTS (comNTS), medial part of NTS, wall of the fourth ventricle, ventral border of area postrema and com NTS in midline dorsal to the central canal within brain stem, spinal ganglia $T_2-L_4$ and nodose ganglia. Labeled sensory neurons innervating the intestines except the stomach were observed in spinal ganglia $T_6-L_4$. The most labeled sensory neurons from the small intestine to large intestine came from middle thoracic spinal ganglia to upper lumbar spinal ganglia.
Inflammatory process leads to the well-known mucosal damage and therefore a further disturbance of the epithelial barrier function, resulting abnormal intestinal wall function, even further accelerating the inflammatory process[1]. Despite of the records, etiology and pathogenesis of IBD remain rather unclear. There are many studies over the past couple of years have led to great advanced in understanding the inflammatory bowel disease(IBD) and their underlying pathophysiologic mechanisms. From the current understanding, it is likely that chronic inflammation in IBD is due to aggressive cellular immune responses including increased serum concentrations of different cytokines. Therefore, targeted molecules can be specifically eliminated in their expression directly on the transcriptional level. Interesting therapeutic trials are expected against adhesion molecules and pro-inflammatory cytokines such as TNF-${\alpha}$. The future development of immune therapies in IBD therefore holds great promises for better treatment modalities of IBD but will also open important new insights into a further understanding of inflammation pathophysiology. Treatment of cytokine inhibitors such as Immunex(Enbrel) and J&J/Centocor(Remicade) which are mouse-derived monoclonal antibodies have been shown in several studies to modulate the symptoms of patients, however, theses TNF inhibitors also have an adverse effect immune-related problems and also are costly and must be administered by injection. Because of the eventual development of unwanted side effects, these two products are used in only a select patient population. The present study was performed to elucidate the ability of TNF-${\alpha}$ antibodies produced in sheep colostrums to neutralize TNF-${\alpha}$ action in a cell-based bioassay and in a small animal model of intestinal inflammation. In vitro study, inhibitory effect of anti-TNF-${\alpha}$ antibody from the sheep was determined by cell bioassay. The antibody from the sheep at 1 in 10,000 dilution was able to completely inhibit TNF-${\alpha}$ activity in the cell bioassay. The antibodies from the same sheep, but different milkings, exhibited some variability in inhibition of TNF-${\alpha}$ activity, but were all greater than the control sample. In vivo study, the degree of inflammation was severe to experiment, despite of the initial pilot trial, main trial 1 was unable to figure out of any effect of antibody to reduce the impact of PAF and LPS. Main rat trial 2 resulted no significant symptoms like characteristic acute diarrhea and weight loss of colitis. This study suggested that colostrums from sheep immunized against TNF-${\alpha}$ significantly inhibited TNF-${\alpha}$ bioactivity in the cell based assay. And the higher than anticipated variability in the two animal models precluded assessment of the ability of antibody to prevent TNF-${\alpha}$ induced intestinal damage in the intact animal. Further study will require to find out an alternative animal model, which is more acceptable to test anti-TNF-${\alpha}$ IgA therapy for reducing the impact of inflammation on gut dysfunction. And subsequent pre-clinical and clinical testing also need generation of more antibody as current supplies are low.
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