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.

We examined the effect of Bulnesia sarmienti (BS) water extract on hyperlipidemia induced by a high-fat diet. ICR mice were fed a high-fat diet ad libitum for four weeks. Simultaneously, BS water extract was administered intragastrically at 0 mg/kg (control), 30 mg/kg, or 300 mg/kg once daily for four weeks. Male ICR mice were divided into four groups; normal control group (NC), high-fat diet+vehicle treatment group (HF), high-fat diet+BS of 30 mg/kg treatment group (HF+BS30), and high-fat diet+BS of 300 mg/kg treatment group (HF+BS300). The levels of serum biochemical parameters and histological appearances were evaluated. After four weeks, body weight gain and serum levels of triglycerides, total cholesterol, and Low-density lipoprotein (LDL)-cholesterol were significantly higher in the HF group than in the normal control group. Together, serum High-density lipoprotein (HDL)-cholesterol level in the HF group was lower than that in the normal control group. However, treatment with BS resulted in significantly reduced body weight gain and levels of serum triglycerides, total cholesterol, and LDL-choleterol. In addition, serum HDL-cholesterol level in the BS treatment group was significantly elevated, compared to that of the HF group. Histopathological evaluation of the liver showed fat accumulation and swelling of hepatocytes in the high-fat diet group; these abnormalities were ameliorated by treatment with BS. These results suggest that treatment with BS water extract resulted in dose-dependent prevention and mitigation of high-fat diet-induced hyperlipidemia.

In vivo redox reaction is involved in the processes of oxidative diseases. Thus, direct and non-invasive measurement of in vivo free radical reactions in living animals should be essential to understanding the roles of free radicals in pathophysiological phenomena. Electron spin resonance (ESR) technique has been utilized in analysis of free radicals, which are generated through imbalance of in vivo redox status. In vivo ESR/spin probe technique using nitroxide radicals as spin probes was developed for estimation of in vivo free radical reactions in whole living animals. This technique using a probe may become a powerful tool for use in clarifying mechanisms of disease and in monitoring pharmaceutical therapy. The application of ESR was introduced and discussed in this article.

Cutaneous vasculitis is an inflammatory necrotizing disease of the blood vessel walls. In most cases, the etiology is due to immune-complex disease from type III hypersensitivity. Clinical signs include swelling, erythema, purpura, erythematous plaques, ulceration, and necrosis. Clinical lesions may be localized to distal extremities or areas of less extensive collateral circulation. In this case, history, physical examination, laboratory findings, cytology, histopathologic examination, and immunohistochemistry might aid in differentiation of a clinically similar disease. Consequently, this case was diagnosed as cutaneous vasculitis and treated with ointment containing corticosteroid and antibiotics over a period of two weeks. After treatment, all of the clinical lesions had disappeared.

Chronic inflammatory diseases such as Crohn′s disease and ulcerative colitis are associated with increased risk of colon adenocarcinoma. Apoptic induction of colon cancer cells by cytokines and death receptors is an important anti-cancer therapy. We observed that co-administration of TNFα and IFNγ in human colon cancer cell line, HCT116, resulted in cell death and expression of IL-32. Cleavage forms of caspase-3, caspase-9, and PARP were increased in TNFα / IFNγ-treated HCT116. mRNA expression of death receptors, including TNFR1 and Fas were not changed and NO generation was not induced by combination of TNFα and IFNγ. However, mRNA expression of IL-32α, β, and γ was increased in TNFα / IFNγ-treated HCT116. To determine the effect of IL-32 in HCT116 cell apoptosis by TNFα / IFNγ stimulation, IL-32 siRNA-transfected HCT116 cells were cultured with TNFα / IFNγ and cell proliferation was measured. IL-32 siRNA induced slight recovery of cell viability of TNFα / IFNγ-stimulated HCT116. These results suggest that IL-32 is not directly related to apoptosis of HCT116 by TNFα / IFNγ stimulation. However, IL-32 expression by TNFα or TNFα / IFNγ in a colon cancer cell line is very interesting because of the unknown effect of IL-32 in colon cancer. Our study will contribute to development of studies for IL-32 function in human colon cancer and anti-cancer therapies using cytokines.

This study was conducted in order to investigate the effect of black sesame oil on hair growth in a shaving animal model of C57BL/6 mice. Five-week-old male mice were acclimated for one week under 22±1 µl room temperature, 50±5% relative humidity, and 12 hours of a light/dark cycle before beginning the experiment. The animals were divided into three groups, including the normal group (saline, N), positive control group (3% MXD, PC), and experimental group (black sesame oil, E) and received topical application of 100 µl once per day, six days per week, for a period of three weeks. Hair regrowth was evaluated by gross and histological examination. In addition, immunohistochemical observation for SCF, the activities of enzymes, including ALP and γ-GT, and the expression quantity of growth factors, including IGF-1 in the skin of mice was performed or evaluated. Topical treatment with black sesame oil and 3% minoxidil for three weeks to dorsal skin resulted in more rapid acceleration of hair regrowth in the E and PC groups than in the N group. Development and elongation of hair follicles were promoted in the E and PC groups, compared with the N group. Serum ALP activity was significantly higher in the E group, compared with the N group within three weeks (p<0.05). Skin ALP activity was significantly higher in the PC group, compared with the N group within two weeks (p<0.05), and higher in the E and PC groups than in the N group within three weeks with no significant differences. Serum γ-GT activity was significantly higher in the PC group, compared with the N group within two weeks (p<0.01), and significantly higher in the E group, compared with the N group within three weeks (p<0.05). Skin γ-GT activity was significantly higher in the PC group, compared with the N group within two weeks (p<0.05), and significantly higher in the PC (p<0.01) and E (p<0.001) groups, compared with the N group within three weeks. IGF-1 expression in the skin was significantly higher in the PC and E groups, compared with the N group (p<0.01). SCF antigens were heavily stained in bulge, stem cells, and dermal papilla and middle stained in inner root sheath, outer root sheath, and epidermis in the E and PC groups. These results indicate that black sesame oil effectively stimulated hair growth in an animal model via several mechanisms and that it can be used practically for hair growth or prevention of hair loss in human beings.

Microbial lipopolysaccharide (LPS) is an endotoxin conveying the surface receptor complex of toll-like receptor 4 (TLR4)-myeloid differentiation 2 (MD-2) in innate immune cells through ancillary proteins such as LPS-binding protein and CD14. However, TLR4 alone is not sufficient for recognition of LPS. MD-2 is essential for sensing the lipid A domain of LPS and for triggering LPS-induced TLR4 activity across the plasma membrane. Therefore, lipid A domain and its binding to MD-2 are potential drug targets for intervention in endotoxemia as well as other disorders associated with LPS etiology. Here, we reviewed MD-2 as a drug target focused on drug candidates-targeting TLRs, transport of microbial LPS into TLR4/MD-2, crystal structure of TLR4/MD-2 alone, crystal structure of TLR4/MD-2 with bound LPS, lipid A derivatives as MD-2 antagonist, non-lipid antagonists of LPS binding to MD-2, and human disorders-implicated with TLR4/ MD-2. This review could be helpful to understanding the biology of TLR4/MD-2, and suggests the importance of MD-2 as a therapeutic target against inflammatory diseases due to infection.

Epithelial ovarian tumors appear to arise from the ovarian surface epithelium (OSE), a simple squamous-to-cuboidal mesothelium covering the ovary. Ovarian tumorigenesis is the most frequent cause of cancer death in gynecological malignancies; however the exact mechanism of this disease is not well known. A theory of repeated ovulation which contributes to neoplastic transformation of OSE has been proposed, and the process of healing ruptured OSE may contribute to the disease. Therefore, it can be assumed that endocrine and autocrine factors may have an influence on ovarian carcinogenesis in women. Thus, in this review, we suggest that these endocrine and autocrine factors may play a role in ovarian tumorigenesis in regulation of growth-stimulation or -inhibition and/or apoptosis of normal and neoplastic OSE cells via their specific receptors.

The discovery of antibiotics has helped to save the lives of an uncountable number of people. Antibiotics have been grouped in different classes based on their origin, structure, and mechanism of action. An intrinsic and acquired mechanism of antimicrobial resistance has been identified in many bacterial strains that are of high clinical importance. This has seriously jeopardized the use of antibiotics and has also caused the spread of microbes that are resistant to effective first-choice, or “first-line” drugs. Thus, sensible use of antibiotics and the search for effective alternative measures are of high importance in order to minimize the effect due to existing and emerging antimicrobial resistant microbes.

A ten-year-old female Yorkshire terrier was presented to Veterinary Medical Center, Chungbuk National University with signs of anorexia and cough. Blood electrolytes, complete blood count (CBC), and hormone analysis revealed mild elevated lipase and bile acid, moderate hypercholesterolemia and severe elevated alanine transaminase (ALT), alkaline phosphate (ALP), and gamma-glutamyl transferase (GGT). Survey radiographs revealed the potbelly-shaped abdomen, generalized hepatomegaly, and round and smooth marginated mass in right upper abdomen. On ultrasonography, enlarged gallbladder with combination of kiwifruit-like and stellate pattern consistent with gallbladder mucocele was revealed. On day 113 after diagnosis of gallbladder mucocele, free fluid and the hyperechoic structure mainly in the dorsolateral region of the gallbladder was found, which was consistent with gallbladder rupture. Cholecystectomy was performed after gallbladder rupture.

A four-year-old sexually intact male dog and a seven-year-old sexually intact female dog presenting with heavy tick infestation were tentatively diagnosed with tick paralysis. Both dogs died due to respiratory failure. A number of ticks were observed in the head, ears, neck, trunk, and limbs of each dog. In these cases, a presumptive diagnosis of tick paralysis could be based on the presence of a species of tick known to cause paralysis combined with the absence of any other disease previously. Most ticks were identified as Haemaphysalis longicornis according to morphological features. Tick paralysis in dogs has rarely been reported in Korea.

Iron catalyzes the production of free radicals, which can be related to a variety of pathological events, such as cancer and aging. The effect of dietary iron was investigated on formation of colonic aberrant crypt foci (ACF) induced by azoxymethane in male F344 rats. Animals were fed three different diets, including iron-deficient (3 ppm Fe), iron-normal (35 ppm Fe), and iron-overloaded (350 ppm Fe) diets for eight weeks. During the first and second weeks of the experiment, animals received two subcutaneous injections of azoxymethane (AOM, 15 mg/kg body weight) to induce ACF. After staining with methylene blue, the total numbers of ACF and aberrant crypts (AC) were counted on the colonic mucosa. Analysis of blood and serum was performed using a blood cell differential counter and an automatic serum analyzer. Iron-deficient diets induced a significant decrease in red blood cell counts and the values of hemoglobin concentration, hematocrit, mean corpuscular hemoglobin, and mean corpuscular volume, while an iron-overloaded diet did not affect these values. The iron-overloaded diet induced an increase in deposits of iron in the liver of rats, as determined by the ICP method and Perl’s staining. The numbers of ACF per colon showed a slight increase in iron-overloaded or iron-deficient rats, without statistical significance, compared to iron-normal rats. The number of total AC per colon in iron-overloaded rats was significantly higher than that in iron-normal rats (p<0.05). The number of large ACF (≥ 4 AC per ACF) in iron-overloaded rats was also significantly higher than that in iron-normal rats (p<0.05). These results suggest that dietary iron intake may play an important role in colon carcinogenesis in humans and animals.

This study was conducted in order to evaluate the anti-inflammatory activities of Phellinus linteus (PL) water extract against carrageenan- and cotton pellet-induced acute and chronic inflammation in rats. Animals were divided into four experimental groups for conduct of acute and chronic inflammations studies: the vehicle control group, positive control group (aminopyrine 100 mg/kg), PL 10 mg/kg group, and PL 20 mg/kg group. Oral administration of PL water extract has dose-dependent anti-inflammatory effects against both acute and chronic inflammation models. In carrageenan-induced paw edema, significant inhibition was observed at 0.5 and 1 h in the PL 10 mg/kg group and at 0.5, 1, 2, 3, 4, and 6 h in the PL 20 mg/kg group. In cotton pellet-induced granuolma formation, PL water extract at 10 and 20 mg/kg/day also showed significant inhibition in dry weights of granuloma. Results of the free radical scavenging assay showed that PL has a dose-dependent scavenging activity against 2,2-diphenyl-1-picrylhydrazyl free radicals. The results obtained in this study indicate that this plant material may be used beneficially as an anti-inflammatory agent having antioxidant action.

Due to the similarities of their anatomical morphology and physiology to those of humans, pigs have increasingly become useful models for toxicological and pharmacological studies. This study was conducted in order to evaluate the quality control level through hematological features of minipigs. Thus, we attempted to determine whether there are differences in hematological parameters between male and female minipigs. No significant differences in the levels of erythrocytes, hematocrit, mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), leukocytes (WBC), and serum electrolytes were observed between male and female minipigs. However, leukocytes were higher than reference range in both sexes (26.46±2.06 vs 11.6±5.4 for male; 23.58±3.36 vs 10.0±2.0 for female). In differential leukocyte counts, no significant differences in leukocyte parameters were observed between sexes. However, neutrophils were higher than reference range in both sexes (60.65±13.92 vs 33.1±18.9 for male; 46.84±12.80 vs 15.4±5.6 for female). In serum biochemical analysis, no significant differences in biochemical parameters were observed between male and female minipigs. However, aspartate aminotransferase (AST) and alkaline phosphatase (ALT) were higher than reference range, while alanine aminotransferase (ALP) was lower than reference range. In urine analysis, pH of male and female was 8.3±1.0 and 6.4±1.6, showing a higher value in male minipigs. These data indicate that quality control of minipigs must take precedence in conduct of toxicological and pharmacological studies.

Prostatic calculi are common finding in radiographic examination, however, the true incidence and the clinical significance are not clearly known. We investigated the prevalence of prostatic calculi and the relationship between prostatic calcification and urolithiasis. From a database, 305 consecutive subjects older than 30 years of age who had undergone regular health checks in our hospital from January 2007 to July 2009 were enrolled in this study. Prostatic calculi and urinary stones were confirmed by non-enhanced CT. According to the presence of prostatic calculi, they were divided into two groups and the relationship with urolithiasis was determined using statistical analysis. Among 305 male subjects, prostatic calculi were found in 97 (31.9%), which revealed a linear trend with aging (r=0.928, p=0.023), while asymptomatic upper urinary tract stones were found in 45 subjects (14.9%). A significant difference was observed in mean age (p<0.001), however, no difference was observed in prevalence of diabetes mellitus (DM), hypertension (HT), and body mass index (BMI). In the prostatic calculi group, 24 (24.7%) subjects had previously unknown urolithiasis, while only 21 subjects (10.1%) in the non-prostatic calculi group had urinary stones. The prevalence of urolithiasis was significantly higher in men with prostatic calcification than in those without prostatic calcification (OR=2.92, 95% CI: 1.53-5.58, p=0.001). No significant differences in serum chemistry were observed between the two groups. Among the 45 patients with urolithiasis, no significant differences were observed in the mean stone size according to the presence of prostatic calcification. Our data showed that prostatic calcification was prevalent up to approximately 31.9% and was frequently combined with upper urinary tract stones. These results indicate that evaluation for urolithiasis might be needed in patients with incidentally detected prostatic calcification.

Recently, studies on natural source antioxidants have become increasingly active in various fields. Among analytical methods for assessment of antioxidant activity, the DPPH, ABTS, and TBA assays have become quite popular in natural antioxidant studies. One of the reasons is that these methods are simple to use and highly sensitive. In particular, DPPH and ABTS assays are based on the theory that a hydrogen donor is an antioxidant. The antioxidant effect can be easily evaluated by following the decrease of specific UV absorption and the results are usually expressed in terms of Trolox equivalent antioxidant capacity (TEAC). The TBA assay has been commonly used for measurement of lipid oxidation in some foods and biological systems. Because the colored complex is formed by the TBA-malonaldehyde interaction, the TBA value can be evaluated using a UV spectrophotometer or HPLC. However, the TBA-malonaldehyde complex is very unstable and often is either not detected in many oxidized lipids or is a minor product of secondary oxidation. As a result of these defects, numerous studies are ongoing.

Germinated rough rice (GRR) has many healthy effects such as antioxidant activity, inhibition of chronic inflammatory disease and prevention of vascular disease. In this study, germinated Korean rice sample which was planted in 2008 from Korean Rural Development Administration was used. The purpose of this study was to recognize the effect of the ethanol extract of GRR on the ischemic damage of skeletal muscle. Right femoral artery ligation was performed to induce ischemic condition in mice and the mixed basal diet with the ethanol extract of GRR at various levels (100, 200, and 400 mg/kg diet) was given for 2 weeks. After the mice were sacrificed, the hind limb muscle samples of the mice were fixed, sectioned, stained with Haematoxylin-Eosin (H&E), Masson’s trichrome, and performed for Immunohistochemistry (IHC) for caspase-3. In histopathological findings of H&E and Masson’s trichrome stain, the treatment of GRR at the level of 400 mg/kg diminished fibrosis and necrosis in the muscle compared to the control. The expression of caspase-3 protein was also decreased by the treatment of GRR at the level of 400 mg/kg compared to the control. These results suggest that the ethanol extract of GRR may have a protective effect against muscle damage during the ischemia in skeletal muscle of hindlimb.