Purpose: Lingual nerve (LN) damage may be caused by either tumor resection or injury such as wisdom tooth extraction, Although autologous nerve graft is sometimes used to repair the damaged nerve, it has the disadvantage of necessity of another operation for nerve harvesting. Moreover, the results of nerve grafting is not satisfactory. The nerve growth factor (NGF) is well-known to play a critical role in peripheral nerve regeneration and its local delivery to the injured nerve has been continuously tried to enhance nerve regeneration. However, its application has limitations like repeated administration due to short half life of 30 minutes and an in vivo delivery model must allow for direct and local delivery. The aim of this study was to construct a well-functioning $rhNGF-{\beta}$ adenovirus for the ultimate development of improved method to promote peripheral nerve regeneration with enhanced and extended secretion of hNGF from the injured nerve by injecting $rhNGF-{\beta}$ gene directly into crush-injured LN in rat model. Materials and Methods: $hNGF-{\beta}$ gene was prepared from fetal brain cDNA library and cloned into E1/E3 deleted adenoviral vector which contains green fluorescence protein (GFP) gene as a reporter. After large scale production and purification of $rhNGF-{\beta}$ adenovirus, transfection efficiency and its expression at various cells (primary cultured Schwann cells, HEK293 cells, Schwann cell lines, NIH3T3 and CRH cells) were evaluated by fluorescent microscopy, RT-PCR, ELISA, immunocytochemistry. Furthermore, the function of rhNGF-beta, which was secreted from various cells infected with $rhNGF-{\beta}$ adenovirus, was evaluated using neuritogenesis of PC-12 cells. For in vivo evaluation of efficacy of $rhNGF-{\beta}$ adenovirus, the LNs of 8-week old rats were exposed and crush-injured with a small hemostat for 10 seconds. After the injury, $rhNGF-{\beta}$ adenovirus($2{\mu}l,\;1.5{\times}10^{11}pfu$) or saline was administered into the crushed site in the experimental (n=24) and the control group (n=24), respectively. Sham operation of another group of rats (n=9) was performed without administration of either saline or adenovirus. The taste recovery and the change of fungiform papilla were studied at 1, 2, 3 and 4 weeks. Each of the 6 animals was tested with different solutions (0.1M NaCl, 0.1M sucrose, 0.01M QHCl, or 0.01M HCl) by two-bottle test paradigm and the number of papilla was counted using SEM picture of tongue dorsum. LN was explored at the same interval as taste study and evaluated electro-physiologically (peak voltage and nerve conduction velocity) and histomorphometrically (axon count, myelin thickness). Results: The recombinant adenovirus vector carrying $rhNGF-{\beta}$ was constructed and confirmed by restriction endonuclease analysis and DNA sequence analysis. GFP expression was observed in 90% of $rhNGF-{\beta}$ adenovirus infected cells compared with uninfected cells. Total mRNA isolated from $rhNGF-{\beta}$ adenovirus infected cells showed strong RT-PCR band, however uninfected or LacZ recombinant adenovirus infected cells did not. NGF quantification by ELISA showed a maximal release of $18865.4{\pm}310.9pg/ml$ NGF at the 4th day and stably continued till 14 days by $rhNGF-{\beta}$ adenovirus infected Schwann cells. PC-12 cells exposed to media with $rhNGF-{\beta}$ adenovirus infected Schwann cell revealed at the same level of neurite-extension as the commercial NGF did. $rhNGF-{\beta}$ adenovirus injected experimental groups in comparison to the control group exhibited different taste preference ratio. Salty, sweet and sour taste preference ratio were significantly different after 2 weeks from the beginning of the experiment, which were similar to the sham group, but not to the control group.
The case of a patient with a transfusion-related acute lung injury (TRALI) to whom extracorporeal membrane oxygenation (ECMO) had been applied is reported. A 55-year-old male injured with liver laceration (grade 3) without chest injury after car accident. He received lots of blood transfusion and underwent damage control abdominal surgery. In the immediate postoperative period, he suffered from severe hypoxia and respiratory acidosis despite of vigorous management such as 100% oxygen with mechanical ventilation, high PEEP and muscle relaxant. Finally, ECMO was applied to the patients as a last resort. Aggressive treatment with ECMO improved the oxygenation and reduced the acidosis. Unfortunately, the patient died of liver failure and infection. TRALI is a part of acute respiratory distress syndrome (ARDS). The use of ECMO for TRALI induced severe hypoxemia might be a useful option for providing time to allow the injured lung to recover.
The studies on cryopreserved arterial allograft have been focused on cooling methods, pre-treatment, cryoprotectant agents, and preservation temperature. But recently, several studies have reported that thawing methods also play an important role in the occurrence of macroscopic and microscopic cracks. This study was designed to investigate the cell injury after thawing, using a rabbit model to clarify the effect of thawing methods on cryopreserved arteries. Material and Method: Segments of the rabbit aorta were obtained and divided into 3 groups (n=60) according to whether the specimens were fresh (control, n=20), cryopreserved and rapidly thawed (RT) at 37$^{\circ}C$ (n=20), or cryopreserved and subjected to controlled, automated slow thawing (ST)(n=20). Cell damage was established using the TUNEL method and the morphological changes were also evaluated. Result: In the group that was rapidly thawed, the expression of TUNEL (+) cells increased significantly more than in the slowly thawed group. In addition, the endothelial denudation, microvesicles and edema were significant in the rapidly thawed group compared with those changes in the slowly thawed group. Conclusion: Our study suggests that the rapid thawing method may be one of the major causes of cellular damage and delayed rupture in cryopresewed arterial allografts. The expression of TUNEL (+) cells and structural changes were significantly low in the slowly thawed group, which might have contributed to the improvement of graft failure after transplantation.
It was thought desirable to elucidate the influence of the mammillary bodies upon sexual behavior. Twenty male rats were used, of which 9 rats had their mammillary bodies damaged electrolytically through implanted electrodes (mammillary body group) and 11 rats received the same surgery short of electrolytic damage to the mammillary bodies (operated control group). Their sexual behavior was observed 3 weeks after surgery. Observation was discontinued when an ejaculation was followed by a successive intromission. 1. Mounting without intromission occurred significantly less often in the mammillary body group than in the operated control group. 2. No significant group difference existed with regard to the occurrence of mounting with intromission. However, the value tended to rise in the mammillary body group compared with the value of the operated control group. 3. Mountings with and without intromission tended to occure more often in the mammillary body group than in the operated control group, but the difference was nonsignificant. 4. The ejaculatory latency (time interval between the first intromission and the first ejaculation) and the postejaculatory interval (time interval between an ejaculatory act and the next intromission) tended to decrease in the mammillary body group compared with the values of the operated control group. However, again the difference did not reach significancy. 5. There existed no significant group differences with regard to the mounting latency (time interval between the first encounter with female and the first mounting with or without intromission), the intromission latency (time interval between the first encounter and the first intromission), and the interintromission period (mean time interval between two successive intromissions). From the above results, it may be inferred that the mammillary bodies tend to exert slight and nonsignificant inhibitory influence upon sexual behavior of male rats. However, further study is indicated before reaching final conclusion.
This study was conducted to see whether the hippocampectomy exerted facilitatory influence upon gastric ulceration in animals, and if so, whether the effect of hippocampectomy could be suppressed by adrenalectomy. 107 male rats were divided into 5 groups: rats that had over 90% of their hippocampal tissue removed through an opening on each side of the cerebral cortex(hippocampal group, N=21), rats that received bilateral adrenalectomy(adrenal group, N=29), rats that received adrenalectomy as well as hippocampectomy(hippocampo-adrenal group, N=10), rats that received damage to each side of the cortex over the hippocampus(cortical control group, N=20), and rats that had solely their head skin incised(normal control group, N=27). All rats were kept without restraint or food deprivation until on the 25th day after surgery, the stomach of each rat was inflated with 7ml of physiological saline and then removed under deep anesthesia. The mucosal surface was sketched under dissecting microscope, and enlarged photographs$(4{\times})$ were taken. The percentage of animals developing gastric ulcer in each animal group was calculated, the number of ulcer in each stomach was counted, and the total area of ulceration per stomach was measured on the Photograph with the aid of superimposed graph paper and expressed as permillage of total area of the glandular mucosa. Results obtained were as follows: 1. The percentage of animals developing gastric ulcer was significantly larger in the hippocampal group than they were in the hippocampo-adrenal, the adrenal, the cortical, and the normal control groups. 2. The mean number of ulcer per stomach was significantly larger in the hippocampal group than they were in the adrenal, the cortical control, and the normal control groups, while no significant difference existed between the hippocampal and the hippocampo-adrenal groups. 3. Total area of ulcer per stomach was significantly larger in the hippocampal group than they were in the cortical control and the normal control groups, but no significant differ-ence existed among the hippocampal, the adrenal, and the hippocampo·adrenal groups. 4. All measured values of the adrenal group were not significantly different from those of the hippocampo-adrenal, the cortical control, and the normal control groups. It is inferred from the above results that the hippocampus exerts an inhibitory influence upon gastric ulceration and that the hippocampal influence is mediated only partly through suppression of pituitary·adrenal activity.
Background: Spinal cord ischemic injury during thoracic and thoracoabdominal aortic surgeries remains a potentially devastating outcome despite using various methods of protection. Neuronal voltage-dependent sodium channel antagonists are known to provide neuroprotection in cerebral ischemic models. This study was designed to compare the neuroprotective effects of phenytoin with those of hypothermia in a rabbit model of spinal cord ischemia. Material and Method: Spinal cord ischemia was induced in New Zealand white rabbits by means of infrarenal aortic cross clamping for 25 minutes. Four groups of 8 animals each were studied. The control group and the hypothermia group received retrograde infusion of saline only ($22^{\circ}C$, 2 mL/min); the normothermic phenytoin group and the hypothermicphenytoin group received retrograde infusion of 100 mg of phenytoin at different rectal temperatures ($39^{\circ}C$ and $37^{\circ}C$, respectively) during the ischemic period. The neurologic function was assessed at 24 and 72 hours after the operation with using the modified Tarlov criteria. The spinal cords were harvested after the final neurologic examination for histopathological examination to objectively quantify the amount of neuronal damage. Result: No major adverse effects were observed with the retrograde phenytoin infusion during the aortic ischemic period. All the control rabbits became severely paraplegic, Both the phenytoin group and the hypothermia group had a better neurological status than did the control group (p < 0.05). The typical morphological changes that are characteristic of neuronal necrosis in the gray matter of the control animals were demonstrated by means of the histopathological examination, whereas phenytoin or hypothermia prevented or attenuated these necrotic phenomena (p < 0.05). The number of motor neuron cells positive for TUNEL staining was significantly reduced, to a similar extent, in the rabbits treated with phenytoin or hypothermia. Phenytoin and hypothermia had some additive neuroprotective effect, but there was no statistical significance between the two on the neurological and histopathological analysis. Conclusion: The neurological and histopathological analysis consistently demonstrated that both phenytoin and hypothermia may afford significant spinal cord protection to a similar extent during spinal cord ischemia in rabbits, although no significant additive effects were noticed.
Park, Seongkyu;Minar, Maruf;Hwang, Yawon;Kim, Somin;Park, Minhyeok;Choi, Seok-Hwa;Kim, Gonhyung
Journal of Veterinary Clinics
/
v.30
no.5
/
pp.346-352
/
2013
The study was aimed to investigate the influence of diode laser on osteoarthritis (OA) of stifle joint induced by anterior cruciate ligament transection (ACLT). Sixty 10-week-old male Sprague-Dawley rats were used in this study. Right stifle joint was operated to create ACLT or sham. There were five study groups: control, Sham, ACLT, ACLT + Laser irradiation (ACLT+L) and ACLT + meloxicam administration (ACLT+M). Low-level laser therapy (LLLT) was applied at the operated stifle joint twice a week using an 808-nm indium-gallium-arsenide (InGaAs) diode laser during 8-week experimental period. Radiographical, gross morphological and histopathological findings were examined at 2, 4 and 8 weeks post-surgery. Radiography, CBC and chemistry tests showed no significant difference between groups. ACLT+L group showed remarkable cartilage damages compared with sham group morphologically and histopathologically at 2, 4 and 8 weeks after surgery. ACLT+M group also had more cartilage damages compared with sham group. Low-level laser therapy (LLLT) showed limitation to prevent progression of OA in the rat anterior cruciate ligament transection models; on the contrary it accelerated cartilage damage. It is assumed that the aggravating results of LLLT in this study might be due to excessive unstable movement of stifle joint from the pain-relieving effect of LLLT, rather than direct damaging effect of irradiation since LLLT did not affect cell viability.
Wounds are commonly created during almost every kind of surgery, trauma and skin diseases. Delayed wound healing affects a plenty of patients and requires prolonged treatments that seriously reduce the quality of life for patients. Skin damage involving large areas or great severity can lead to disability or even death. Wound healing involves a complicated series of actions, of various tissues and cell lineages, concerning inflammation, migration, proliferation, reepithelialization, and remodeling. Sopung-san is reported to have anti-inflammatory effect and has been used for various skin diseases such as allergic dermatitis and atopic dermatitis. In this study, the hypothesis that oral treatment with Sopung-san could enhances healing potential on rat full thickness skin wounds was tested. Twenty young male Sprague-Dawley rats were used for the studies. A full-thickness skin wound was made on the dorsal skin of the rats. Either Sopung-san water extract (SPS) or saline (Control) was orally administrated every day. The wound area was measured and the percentages of wound contraction, wound healed and wound epithelization were calculated. Wound tissue samples were excised following injection for histopathological and immunohistological examination. Wound area in rats of SPS group significantly was decreased compared to Control. SPS group showed significant promotion of wound healing compared to Cotrol group in the percentages of wound contraction, wound healed and wound epithelization. Histopathological examination revealed that SPS induces neo-vascularization potential in wound healing process. SPS treatment in rats significantly accelerated cutaneous wound healing in the neo-vascularization process by increasing VEGF and $TGF-{\beta}1$ synthesis. The results suggest that Sopung-san affects key cellular processes responsible for wound repair and point to a unique potential for this molecule in the therapy of skin wounds, particularly as an angiogenic agent.
Objective : In the present study we analyzed neuroprotective and antiapoptotic effect of the difumarate salt S-15176, as an anti-ischemic, an antioxidant and a stabilizer of mitochondrial membrane in secondary damage following spinal cord injury (SCI) in a rat model. Methods : Three groups were performed with 30 Wistar rats; control (1), trauma (2), and a trauma+S-15176 (10 mg/kg i.p., dimethyl sulfoxide) treatment (3). SCI was performed at the thoracic level using the weight-drop technique. Spinal cord tissues were collected following intracardiac perfusion in 3rd and 7th days of posttrauma. Hematoxylin and eosin staining for histopatology, terminal deoxynucleotidyl transferase dUTP nick end labeling assay for apoptotic cells and immunohistochemistry for proapoptotic cytochrome-c, Bax and caspase 9 were performed to all groups. Functional recovery test were applied to each group in 3rd and 7th days following SCI. Results : In trauma group, edematous regions, diffuse hemorrhage, necrosis, leukocyte infiltration and severe degeneration in motor neurons were observed prominently in gray matter. The number of apoptotic cells was significantly higher (p<0.05) than control group. In the S-15176-treated groups, apoptotic cell number in 3rd and 7th days (p<0.001), also cytochrome-c (p<0.001), Bax (p<0.001) and caspase 9 immunoreactive cells (p<0.001) were significantly decreased in number compared to trauma groups. Hemorrhage and edema in the focal areas were also noticed in gray matter of treatment groups. Results of the locomotor test were significantly increased in treatment group (p<0.05) when compared to trauma groups. Conclusion : We suggest that difumarate salt S-15176 prevents mitochondrial pathways of apoptosis and protects spinal cord from secondary injury and helps to preserve motor function following SCI in rats.
Lee, Jin A;Oh, Min Hyuck;Shin, Mi-Rae;Roh, Seong-Soo;Park, Hae-Jin
Korean Journal of Pharmacognosy
/
v.52
no.3
/
pp.177-185
/
2021
Gastroesophageal reflux disease (GERD) is a gastrointestinal disorder in which stomach contents reflux into the esophagus, causing complications such as mucosal damage. The purpose of this study was to determine the effect of Rhei Rhizoma and Scutellariae Radix mixture (RS) in chronic acid reflux esophagitis (CARE), one of the GERD. After inducing reflux esophagitis through surgery, the group was separated and the drug was administered for 2 weeks; Normal rats (Normal, n=8), chronic acid reflux esophagitis rats (Control, n=8), tocopherol 30 mg/kg-treated chronic acid reflux esophagitis rats (Toco, n=8), Rhei Rhizoma and Scutellariae Radix mixture 100 mg/kg-treated chronic acid reflux esophagitis rats (RSL, n=8), Rhei Rhizoma and Scutellariae Radix mixture 200 mg/kg-treated chronic acid reflux esophagitis rats (RSH, n=8). Gross lesion of esophageal mucosa after RS treatment showed a superior enhancement compared with that of Control group. Additionally, RS significantly decreased the levels of MPO and MDA, effectively inhibited NADPH oxidase, and regulated the expression of the AMPK/LKB1/NF-κB pathway. Moreover, it significantly increased the expression of tight junction proteins. Taken together, RS not only alleviates inflammation of the esophageal mucosa via the AMPK/LKB1/NF-κB pathway by reducing oxidative stress, but also improves esophageal function by modulating tight junction proteins.
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