Liver cancer represents a major health problem with steadily increasing incidence rates. Hepatocellular carcinoma (HCC) is the fifth most common cancer and the third most common cause of cancer-related death. This study was conducted in order to investigate the gross findings following treatment with diethylnitrosamine (DEN) in mice. Sixteen male and female mice (B6C3F₁), initially 20 days of age, received intraparietal injection (20 mg/kg three times for a period of two weeks, IP) or were given drinking water (DW) containing 50 ppm DEN; all mice were sacrificed at the 80^th week of experiments. Hepatocellular adenoma (HCA) and HCC were induced in B6C3F₁), mice by administration of DEN. The numbers of HCA and HCC were 7.4±1.72 (IP) and 7.2±0.86 (DW) in male mice. However, no significant difference was observed between the DW and IP groups. The numbers of HCA and HCC were 0.67±0.33 (IP) and 2.0±0.63 (DW) in female mice. This study showed a tendency for high incidences of liver tumor with long-term exposure of newborn animals by drinking water.

Peripheral nerve injuries are very common in clinics and often result in severe functional deficits. The aim of this study was to evaluate the effect of treadmill running and electro-acupuncture on nerve regeneration and functional recovery of muscle activity following sciatic nerve crush injury in a rat model. A comparative study was conducted over 30 days on 60 adult male Sprague-Dawley rats grouped into sham control (C), electro-acupuncture (EA), treadmill (T), and treadmill plus electro-acupuncture (TEA). The left sciatic nerve was crushed for 30 sec using a hemostatic forceps and functional activity was evaluated with sciatic functional tests, nerve conduct velocity, muscle weight, and histology at 10, 20, and 30 days after injury. Muscle weight was significantly (P<0.05) increased between days 10 and 30 in the TEA group. In histology, the degree of damage was scored as C > TEA > T > EA, although necrosis and fibrosis of muscle was observed only in the TEA group. The EA and TEA groups showed rapid recovery with better myelinated axons on day 10. These results suggest that application of the TEA method with balanced exercise is a useful treatment option for peripheral nerve injury regeneration and muscle activity.

In all living organisms, respiration may lead to oxidative stress, a state where increased formation of reactive oxygen species overwhelms host protection and subsequently induces DNA damage, lipid peroxidation, and protein denaturation. As a phenolic acid, chlorogenic acid occurs ubiquitously in food. It has been proven to have a number of biological effects in vitro and in vivo, including antioxidant, anti-inflammatory, and anticarcinogenic properties. Therefore, to some extent, chlorogenic acid can promote human health, and hopefully provide new methods for treatment of chronic disease. Recent studies have focused on the antioxidant properties of dietary polyphenols. The in vitro data often conflict with results obtained from in vivo studies on the antioxidant capacity of plasma or the resistance of plasma and lipoproteins to oxidation ex vivo after consumption of polyphenol-rich foods by human subjects. These inconsistencies are likely explained by the limited bioavailability of dietary polyphenols and their extensive metabolism in humans. Polyphenols exert multifaceted actions, and any clinical application using these substances should be based on a precise understanding of the physiologically relevant mechanisms.

Canine heartworm disease is typically characterized by the presence of heartworms in the pulmonary artery and, to a lesser degree, the right ventricle. Systemic arterial dirofilariasis is an unusual manifestation of heartworm disease of dogs that results from aberrant migration of Dirofilaria immitis into the peripheral arterial circulation. A three-year-old Jindo dog was referred to the Veterinary Medical Center because of hindlimb lameness, paresthesia of hindlimbs, and interdigital ischemic necrosis. Angiography and ultrasonography were performed, resulting in diagnosis of heartworm infestation. In necropsy, entangled heartworms were present throughout the body, including lungs, kidney, small intestinal lumen, and arteries, in addition to the right ventricle and pulmonary artery. Histologically, lung showed adult heart worms in vessels, organized thrombotic vessels, and interstitial pneumonia featuring type II pneumocyte hyperplasia, thickened alveolar wall, and compressed alveolar spaces. Chronic parenchymal lesions, such as focal or diffuse necrosis, fibrosis, and dystrophic calcification were remarkable in liver and kidney. The current case showed a typical canine systemic heartworm disease caused by aberrant migration of Dirofilaria immitis.

We investigated free radical reactions in lung of living mice using an in vivo electron spin resonance (ESR) spectrometer and nitroxyl radical as a probe. When an aqueous solution of nitroxyl probe was trans-tracheally administered into lung of living mice, a sharp triplet signal was observed at the chest of the mice. The signal showed a gradual decrease with time, obeying first-order kinetics. Signal decay rates of carbamoyl-PROXYL and carboxy-2,2,6,6-tetramethyl-piperidine-N-oxyl were faster than those of CAT-1 and carboxy-PROXYL. The mechanism of signal decay may be attributed to (i) reaction with reactive oxygen species such as ·OH, (ii) transfer into blood circulation, or (iii) reduction by compounds continuously supplied. However, little is known about the clearance mechanism of the nitroxyl probe in lung. To evaluate the disappearance of the ESR signal of the nitroxyl probe in lung, in vivo ESR spectra in chest of mice was recorded after trans-tracheal administration of an aqueous high concentrate solution of nitroxyl probe. A broad signal from the chest was observed immediately after administration due to Heisenberg spin exchange interaction. A sharp triplet signal was superimposed on the broad signal and the appearance of a triplet signal was followed by disappearance of the broad one. Peak-to-peak line width of the sharp signal was almost the same as that after intravenous administration. A distinct signal was detected in blood collected 10 min after trans-tracheal administration of nitroxyl probe. The observations indicate the transfer from lung to blood circulation and its contribution to clearance of probe in lung. Appearance of a sharp signal in blood after trans-tracheal administration was dependent on the kind of nitroxyl probe, showing a different transfer rate from lung to blood.

To investigate the effect of carnosine on exhaustive exercise, swimming tests were conducted weekly with loads corresponding to 5% of body weight attached to the tails of mice, and the swimming time to exhaustion was measured. Eighty male ICR mice were divided into four groups, to which carnosine was administered at doses of 0 (control), 10, 50, and 250 mg/kg/day, respectively, for a period of four weeks. At the end of swimming exercise challenges, serum biochemistry, oxidative stress enzyme activity, and antioxidant enzyme activity in tissues were determined. Treatment with 250 mg/kg carnosine resulted in a significant increase in swimming times to exhaustion, compared to the control group in the first (P<0.01) and third week (P<0.05). Significantly lower serum lactate levels were observed after the swimming exercise in the carnosine-treated groups (10 and 250 mg/kg), compared with the control (P<0.01). Malondialdehyde levels in the liver (10 and 50 mg/kg carnosine treated groups) and skeletal muscle (250 mg/kg carnosine treated group) were significantly lower, compared with the control (P<0.05). Significantly lower protein carbonyl levels in skeletal muscle were observed in the 50 and 250 mg/kg carnosine treated groups, compared with the control (P<0.01). Superoxide dismutase and glutathione peroxidase activities in skeletal muscle did not differ significantly among the groups. These results indicate that carnosine may improve swimming exercise capacity by attenuating production of lactate and reducing oxidative stress in mice.

Neurotoxicity and oxidative injury induced by glutamate cause neuronal degeneration related to various central nervous system diseases. Resveratrol, a polyphenolic compound, is known to have antioxidative and anti-inflammatory effects. The aim of this study was to investigate the question of whether resveratrol has a neuroprotective effect against glutamate-induced toxicity in cultured cortical neurons. Following exposure to glutamate for 15 min, cortical neurons originating from ICR mouse fetuses on embryonic days 15-16 were then treated with resveratrol for 24 h in the post-treatment paradigm. Glutamate induced a significant reduction in cell viability; however, resveratrol induced a significant increase in cell viability. Glutamate induced generation of ROS and apoptotic neuronal death; however, these were decreased by exposure to resveratrol. mRNA expression in antioxidant enzymes, cytoplasmic glutathione peroxidase, copper/zinc superoxide dismutase (SOD), and manganese SOD, and anti-apoptotic regulator Bcl-xL were decreased by exposure to glutamate, however, exposure to resveratrol resulted in a significant increase in their mRNA levels. In addition, mRNA expression of pro-inflammatory cytokines, interleukin-1β and tumor necosis factor-α, was increased by glutamate insult, but significantly reduced by resveratrol. These findings indicate that resveratrol is neuroprotective against glutamate-induced toxicity, suggesting a useful therapeutic application in treatment of neurodegenerative disorders.

Effects of carnitine on atherosclerosis and steatosis of hypercholesterolemic rabbits induced by a high-cholesterol diet (HCD) containing 0.5% cholesterol and 2.0% corn oil were investigated. Male New Zealand white rabbits with hypercholesterolemia (blood cholesterol 1,000-1,500 mg/dl) induced by two-week feeding a HCD, were fed a HCD containing 0.008 or 0.075% L-carnitine for an additional eight weeks. Feeding a HCD for 10 weeks resulted in severe atheromatous change, covering 55.7% of the aortic walls, in addition to profound hepatic steatosis. However, carnitine supplementation resulted in recovery of the increased low-density lipoproteins and triglycerides and a decrease in the levels of high-density lipoproteins following HCD feeding, although the increased cholesterol concentration was not potentially attenuated. Notably, carnitine induced a marked reduction of the atheroma area and hepatic lipid accumulation as well as lipid peroxidation. The results of this study indicated that carnitine exerted anti-atherosclerotic and fatty liver-preventing activities through blockade of lipid peroxidation and regulation of lipid metabolism.

Sepsis is a clinical syndrome defined as a systemic inflammatory response to infection. Eritoran is a synthetic lipid A derivative that competes with lipopolysaccharide in binding to the identical site of myeloid differentiation-2/toll-like receptor 4 complex. Eritoran is effective in decreasing the septic mortality of Gram-negative bacteria-infected animals. Eritoran has been highlighted as a candidate drug for treatment of endotoxemia in phase I clinical studies with healthy human volunteers. A phase II trial of eritoran has been conducted in patients with severe sepsis. Intravenous infusion of eritoran reduced the mortality rate, as compared with the placebo group, in sepsis patients at a high risk of mortality according to acute physiology and chronic health evaluation-II scores. A phase III study of eritoran was completed in 2011. The results appear to be disappointing as no statistically significant difference in all-cause mortality was observed between the eritoran treatment group and the placebo group on day 28 in sepsis patients with a high risk of death. In this review, we focus on the rationale for the use of eritoran in treatment of sepsis as well as its clinical applications.

A nine-month-old male Pekingese weighing 5.7 kg was admitted to the Veterinary Medical Center at Chungbuk National University with a history of acute nonambulatory tetraparesis after minor trauma. A diagnosis of atlantoaxial instability with a dens axis fracture was based on examination of survey spinal radiographs and was confirmed during surgery. A modified ventral fixation technique using cortical screws was used for stabilization of the atlantoaxial joint. Serial evaluation based on radiographic and neurologic assessment was performed eight weeks after surgery. Symptoms of tetraparesis disappeared gradually, and arthrodesis of the atlantoaxial joint using a ventral fixation technique has maintained stable fixation.

Phospholipid hydroperoxide glutathione peroxidase (PHGPx) is a unique antioxidant enzyme involved in reduction of peroxidized phospholipids within biomembranes. To investigate the expression pattern of the PHGPx gene during fetal development, in situ hybridization analyses were performed using mouse FITC-labeled PHGPx cRNA probes in fetal tissues on embryonic days (Ed) 13.5-18.5. During these periods, PHGPx mRNA appeared in the developing telencephalon, diencephalon, spinal cord, and spinal ganglion. In particular, PHGPx mRNA was strongly expressed in pyramidal cells of the cerebral cortex. On Eds 17.5-18.5, PHGPx mRNA was detected in various tissues including liver, intestinal villi and crypt, pancreas, lung, and olfactory epithelium of the nasal cavity. In addition, PHGPx mRNA was highly expressed in the inner ear on Eds 14.5-18.5, brown fat on Ed 17.5, and adrenal gland on Ed 18.5. It is conceivable that PHGPx may act as an important antioxidant against fetal oxidative stress during mouse organogenesis.

For application of nano-sized material in various fields, toxicity evaluation of nano-sized material is important. In the current study, a suspension of 50 nm-sized zinc oxide (ZnO) nanoparticles at a dose of 1 g/kg body weight was injected intraperitonially into mice in order to identify the toxicity of ZnO nanoparticles. After 24 h, the blood and liver were taken and analyzed. According to the results of hematological analysis, white blood cell (p<0.001), mean corpuscular hemoglobin (p<0.001), and mean corpuscular hemoglobin concentration (p<0.05) in the ZnO nanoparticle treated group showed a significant decrease, compared to the control group. In serum biochemistry analysis, alanine aminotransferase (p<0.001) and aspartate amino-transferase (p<0.05) also induced a significant increase in the ZnO nanoparticle treated group, compared with the control group. In the histopathological examination, liver in mice treated with ZnO nanoparticles showed edema and degeneration in hepatocytes. Therefore, it is concluded that the liver is the target organ for 50 nm ZnO intraperitoneal exposure. In the future, greater attention should be paid to the potential toxicity induced by various routes and doses of ZnO nanoparticles.

The effects of isotonic saline on corneal penetration, thickness, and injury, as well as lacrimal secretion in a rat model of dry eye were investigated, in comparison with distilled water. Male Sprague-Dawley rats received intraperitoneal administration of atropine sulfate (20 mg/kg) and their eyes were exposed to dry (relative humidity 25-35%) air flow (2.4 m/sec), under Zoletil anesthesia, for 5 hr to induce dry eye. During the period of dry eye induction, distilled water or isotonic saline (5 μl) was instillated onto the cornea every 30 min. Corneal penetration was measured through fluorescein dye quantification, and corneal thickness and injury were examined under a microscope. Lacrimal (tear) secretion and mucin-like glyocoprotein excretion from goblet cells were measured using the Schirmer test and microscopy, respectively. In dry eye rats treated with distilled water, corneal thickness, tear secretion, and mucin-like glycoprotein excretion were decreased to 74.0%, 74.1%, and 46.3% of normal levels, respectively, resulting in marked corneal injury and a significant increase in corneal penetration. In comparison, treatment with isotonic saline resulted in recovery of lacrimal secretion, in spite of a slight improvement of mucin-like glycoprotein excretion, and thereby prevented corneal penetration of fluorescein by 10%. The results indicate that repeated instillation of isotonic saline could provide slightly greater protection from corneal injury, compared with distilled water by facilitating lacrimal secretion, in addition to relief of inconvenience and pain.

The aim of this study was to evaluate the alleviative effects of a nude pack containing black tea water extract (NPBT) on inflammation and skin barrier damage in hairless mice. Skin inflammation and skin barrier damage were induced by UVB irradiation to the backs of hairless mice for five weeks. And, at the same time, saline (C), NPBT (1%: E1, 2%: E2), and 0.01% retinoic acid diluted with polyethylene glycol (PC) were applied topically twice per day, five days per week, for a period of five weeks. The skin erythema indices of the E1 and E2 groups were significantly lower (p<0.05) than those of the C group through one week after the beginning of the experiment. Meanwhile, water contents of the E1 and E2 groups were significantly higher (p<0.05) than those of the C group through two weeks after the beginning of the experiment. Remarkable thickening of epidermis and dermis was observed in the C group, compared with the PC, E1, or E2 groups. In the C group, infiltration of many inflammatory cells, including neutrophils and lymphocytes, was observed in dermis and a large number of mast cells were observed in dermis and hypodermis, and the degree of degranulation was remarkable. However, these phenomena were alleviated in the PC, E1, and E2 groups. Therefore, NPBT could have considerable inhibitory efficacy on inflammation and skin barrier damage induced by UVB irradiation.

Influenza virus infection is a zoonosis that results in high mortality in animals and humans. Several recent avian influenza outbreaks have posed a significant public health threat to humans because there have been many cases of direct interspecies transmission from birds to humans. Influenza virus infection causes acute respiratory failure, which is the main cause of death, while chronic influenza virus infection in the central nervous system results in neural dysfunction. Of particular interest, according to one report, a group of patients who recovered from influenza virus infection during pandemics showed neuropsychiatric disorders, including depression, schizophrenia, and Parkinsonism. Thus, study of the etiology of neuropsychiatric disorders caused by influenza virus infection is needed. In order to conduct necessary experiments, it is essential to reproduce neurological phenomena that are manifested by human patients who have recovered from influenza virus infection using laboratory animals. In this review, we will discuss some of the facts that should be considered when establishing an animal model for study of central nervous system responses to influenza virus infection using mice.

A six-year-old male Pekinese dog was referred to the Veterinary Medical Center, Chungbuk National University because of intermittent vomiting, anorexia, alopecia, and pruritus. Generalized alopecia, pigmentation, and papular erythema in the skin were observed on physical examination. Hematological studies indicated a severe pancytopenia and hypoalbuminemia. A hormone analysis indicated a hyperestrogenemia. A circumscribed mass, measuring approximately 3-4 cm in diameter, was observed on abdominal radiographs. Grossly, the cryptorchid testis was enlarged by a firm and white spherical mass, measuring approximately 3 cm in diameter, which, on its cut surface, was creamy white with hemorrhage. The normal scrotal testis was markedly atrophic and soft. Histologically, the intra-abdominal cryptorchid testis contained abundant fibrous tissue stroma and the stromal tissues were arranged in a tubular pattern in which the neoplastic cells tended to palisade. The cells had poor and pale eosinophilic cytoplasm streaming into the center of the tubules. The nuclei were round to oval and tended to be in a basilar location. Regions of hyperplasia and marked squamous metaplasia were observed in many areas of the prostate. Based on the histological findings, we were able to identify these masses as Sertoli cell tumors, and made a final diagnosis as Sertoli cell tumors through immunohistochemistry methods using inhibin-α, vimentin, and neuron-specific enolase.

This study was conducted in order to investigate repeated-dose toxicities of Magnolia ovobata ethanol extract (MEE). MEE was administered orally to male and female Sprague Dawley rats at dose levels of 0, 500, 1,000, or 2,000 mg/kg for four weeks. Repeated administration of MEE did not induce abnormalities in general signs, body weight gain, feed and water consumption, necropsy findings, or organ weights. In addition, no abnormality was observed in hematological analyses; red blood cells and their indices, white blood cells, platelets, and coagulation times. In male rats, BUN and creatinine showed an increase at doses of 2,000 mg/kg and 500-1,000 mg/kg, respectively, while in female rats, lactate dehydrogenase and creatine phosphokinase showed a decrease at 2,000 mg/kg, the upper-limit dose of repeated-dose toxicity studies. However, there were no dose-dependent increases or gender-relationship. In addition, other parameters of the hepatic and muscular toxicities as well as energy and lipid metabolism were not affected. In microscopic examination, no considerable pathological findings were observed. The results indicate the safety of oral administration of MEE to the upper-limit dose.

Animal crude drugs (natural medicines derived from animal organs) have been widely used in various Chinese medicine for the therapeutic effects and for enhancement of immunologic functions. We found the specific identification methods using DNA sequencing and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analyses for mitochondrial DNA (mt DNA) in order to discriminate between the animal species and organs as well as the placenta of humans. Species-specific PCR bands of D-loop mt DNA for equine, bovine, porcine, and human were 133 bp, 137 bp, 231 bp, and 240 bp, respectively. Porcine organs were identified using restriction enzyme, HphI cut into two subfragments, 36 bp and 195 bp bands in the heart, spleen, and liver, except for kidney. The heart and liver of porcine were identified using restriction enzyme, SpeI cut into two subfragments, 84 bp and 147 bp bands, except for kidney and spleen. Bovine organs were cut into 68 bp and 69 bp bands in the liver, kidney, and spleen using NalIV, except heart and placenta. Placentas of bovine and humans were easily identified using each primer. Our results suggest that sequencing of mt DNA and its PCR-RFLP methods are useful for identification and discrimination of inter- and intra-specific variations in equine, bovine, porcine, and human by routine analysis.

Atherosclerosis, a disease of the large arteries, is the primary cause of heart disease and stroke. The abnormal proliferation of vascular smooth muscle cells (VSMCs) in arterial walls is an important pathogenetic factor of vascular disorders like atherosclerosis and restenosis after angioplasty. In the current study, the possible anti-proliferative effect of vitexin originated from Acer palmatum was investigated in rat aortic VSMCs. Vitexin was found to potently inhibit 5% fetal bovine serum (FBS)-induced growth of VSMCs. Pre-treatment with vitexin (5-50 μg/ml) in VSMCs for 24 h resulted in a significant decrease in cell number, i.e., the inhibition rates were 5.4±7.1, 52.5±8.4, and 78.9±5.2% with vitexin treatments of 5, 20, and 50 μg/ml, respectively. In addition, trteatment with vitexin resulted in significant and dose-dependent inhibition of 5% FBS-induced DNA synthesis. Vitexin did not show any cytotoxicity in rat aortic VSMCs under this experimental condition. These results indicate the potential for development of vitexin as an anti-proliferative agent for treatment of angioplasty restenosis and atherosclerosis.

Wrinkles are an outward sign of cutaneous aging appearing preferentially on ultraviolet B (UVB)-exposed areas. The anti-wrinkle effects of herbal extracts were investigated in an animal model. Female albino hairless mice (HR/ICR) were randomly allocated to the control group (non-irradiated vehicle), positive control group (UVB irradiated-vehicle), and two herbal extract mixture groups (HE-1 and HE-2). HE-1 included Glycyrrhizae radix, Rhei Rhizoma, Cornus officinalis, and Sesami semeni, and HE-2 included Swertia pseudo-chinensis, Sophora flavescens, Scutellaria baicalensis, and Salvia miltiorrhiza. The herbal extract mixtures were pre-treated dorsally with 0.2 ml per individual five times per week for four weeks prior to the start of UVB irradiation. At the fifth week, the animals were exposed to UVB irradiation for a subsequent eight weeks, three times per week. The intensity of irradiation showed a gradual increase, from 30 mJ/cm 2 to 240 mJ/cm2 (1 MED: 60 mJ/cm2 ). Dorsal skin samples were stained with H&E in order to examine the epidermal thickness. In addition, Masson-Trichrome staining was performed for determination of the amount of collagen fiber. Treatments with HE-1&2 resulted in an increase in the amount of collagen fiber, a better appearance, and fewer wrinkles, compared with the positive control. As determined by hydroxyproline assay, treatments with HE-1&2 led to a significant increase in the amount of collagen, compared with the positive control group (p<0.05). Chronic UVB irradiation to skin of hairless mice resulted in an increase in expression of matrix metalloproteinase-1 (MMP-1), however, treatments with HE-1&2 tended to decrease the expression of MMP-1. These results indicate that the herbal extracts used in this study have a preventive effect on UVB-induced wrinkle formation in a hairless mouse model, due in part to inhibition of MMP-1 expression and increment of collagen amount.