The anti-inflammatory effect of PHBV/Collagen (PHCP) was examined in a mouse model of lipopolysaccharide (LPS)-induced skin inflammation. Vascular permeability on the back skin was measured by the local accumulation of Evan’s blue dye after subcutaneous injection of LPS (30 µg site^-1 ). Dye leakage in the skin showed a significant increase at 2 h after injection of LPS. This LPS-induced dye leakage was also completely inhibited by HO-1 inhibitor, ZnPP, and antioxidants, including methyl gallate, trolox, and mannitol. To study the possible mechanisms underlying the in vivo anti-inflammatory effect of PHCP against LPS-induced inflammation, we also examined the effects of PHCP on malondialdehyde (MDA) and glutathione levels in skin tissues and found that pretreatment with PHCP resulted in inhibited MDA elevation and a remarkable reduction of glutathione level. In addition, similar results were obtained after pretreatment with antioxidants, including trolox and mannitol, and HO-1 inhibitor, ZnPP. Histopathologically, an influx of neutrophils into the skin dermis was detected between 24 h and 72 h after LPS injection (30, 100 µg site^-1), compared to control animals after injection of saline. This increase was greater in mice treated with 100 µg of LPS than in those treated with 30 µg of LPS and was significantly suppressed by pretreatment with PHCP, antioxidants, and HO-1 inhibitor. These results collectively suggest that PHCP has an anti-inflammatory effect against LPS-induced inflammation model in vivo and may be a good candidate for the skin tissue engineering biomedical application primarily through manipulation of the redox state.

This study was conducted in order to examine the safety of bee venom as an alternative for antibiotics using male ICR mice. Five-week-old male mice received a single intravenous injection of a dried honey bee venom at the concentration of 0.25 mg/kg (a clinical dose) or 0.5 mg/kg through the tail vein and various pathophysiological analyses were performed after three days. No significant differences in changes of body weight were observed between the saline-treated control group and the experimental groups. In the hematological analysis, none of the parameters were affected by bee venom. In blood biochemistry analysis, none of the markers were affected by administration of bee venom. Similarly, there were no significant effects on markers for liver, kidney, and skeletal muscle functions in all treated- groups. On macroscopic examination, no remarkable lesions were detected in these organs. Because there were no adverse effects of the bee venom in a single intravenous toxicity test for three days, it was concluded that bee venom could be a candidate for a safe natural antibiotic for use in the animal production industry.

Iron nanoparticles (Fe-NPs) have recently been used for cancer diagnosis and therapy for imaging contrast and drug delivery. However, no study on nutritional bioavailability of Fe-NPs in the body has been reported. Ascorbic acid (AA) is known to aid in absorption of iron in the stomach by reducing Fe (III) to Fe (II). In this study, we investigated the bioavailability of Fe-NPs with AA in iron-deficiency-anemic mice in comparison with non-nano iron particles. Iron-deficient anemia was induced by feeding an iron-deficient diet (4.5 mg Fe/kg) and ocular bleeding from retro-orbital venous plexus for four weeks. Normal control mice were given a normal diet (45 mg Fe/ kg). After induction of anemia in mice, anemic mice received daily oral administration of Fe (40 mg/kg B.W.) + AA (5 g/kg B.W) and Fe-NPs (40 mg/kg B.W) + AA (5 g/kg B.W). After sacrifice, liver and spleen tissues were observed by haematoxylin & eosin stain. Amount of trace iron in liver and upper small intestine was investigated using an inductively coupled plasma-atomic emission spectrometer. Red blood cells (RBC), hematocrit (Hct), hemoglobin (Hb), and total iron binding capacity were also measured. The concentrations of iron in the Fe-NPs + AA group were significantly higher in liver and in upper small intestine than that in the Fe + AA group. The values of RBC, Hct, and Hb in the Fe-NPs + AA group were more rapidly increased to normal values compared with the Fe + AA group with increasing time. These results suggest that Fe-NPs in the presence of AA may be more bioavailable than non-nano Fe in Fe-deficient anemic mice.

Experiments were conducted in order to assess the healing effect of bee venom (BV) cream on full-thickness skin wounds in rabbits. BV cream was compared with silver sulfadiazine (SS) as a topical medicament against a control on experimentally created full-thickness wounds. Two wounds measuring 2 × 2 cm were created bilaterally (four wounds/rabbit) on the dorsolateral aspect of the trunk of seven New Zealand white rabbits. Wound treatments were evenly distributed on four sites, using a Latin square design. The contact layer of wounds was treated with physiological saline (control), SS cream, and BV cream over a period of 28 days. Assessment of wound healing was based on scab hardness, wound exudates, wound area, unepithelialized granulation tissue, and histopathological findings. Topical application of BV and SS creams to wounds resulted in reduced inflammation, debridement of necrotic tissue, and promoted granulation and epithelialization. Wound healing was faster, with statistical significance in BV and SS treatments, compared to the control (P<0.05). Treatment with BV evoked an anti-inflammation effect in a rabbit model. BV cream produced a wound healing effect similar to that of commercially available SS cream. Anti-inflammation effect as a topical treatment with BV cream appears to be better than that with SS cream. These results suggest that topical application of BV cream may be an alternative treatment for full-thickness skin wounds.

This study was conducted in order to evaluate the alleviating effects of Phellodendrin cortex water extract (PCWE) on skin aging in hairless mice via observation of morphogical and histological changes. Skin aging was induced by UVB irradiation and application of squalene monohydroperoxide (Sq-OOH) to the back skin of hairless mice for six weeks. And, at the same time, saline (C), jojoba oil (VC), PCWE (E), and 0.01% retinoic acid diluted with polyethylene glycol (PC) were applied topically twice per day, six days per week, for a period of six weeks. Improved wrinkle formation in a pattern of shallow furrows and thin and narrow crests was observed in the retinoic acid and PCWE application groups, compared to the C group. On the morphologic analysis for skin wrinkles, the E group showed lower levels in skin roughness, maximum roughness, average roughness, smoothness depth, and arithmetic average roughness by 13.1, 17.2, 18.4, 15.4, and 16.1%, respectively, compared with the C group, indicating that PCWE inhibited potential formation of wrinkles in the skin. In the C group, structures of lipid lamellae and collagen fibers were broken or deformed with an irregular arrangement. Application of retinoic acid and PCWE protected against the deformity of lipid lamellae and collagen fibers. Elastic fibers in dermis of the C group also showed severe transformation; however, applications of retinoic acid and PCWE resulted in a significant decrease in the number of denatured elastic fibers. Therefore, PCWE could have an alleviating effect on skin aging induced by UVB irradiation and application of Sq-OOH.

Malignant pleural effusion (MPE) and blood samples can be used as a practical source for detection of epidermal growth factor receptor (EGFR) mutations in patients with advanced non-small cell lung cancer. We compared EGFR mutation status of cell blocks, cell-free fluid of MPE, and plasma from patients with lung adenocarcinoma. We obtained paired samples of MPE and plasma from 14 pathologically-confirmed lung adenocarcinoma patients. Peptide nucleic acid (PNA)-mediated real-time polymerase chain reaction (RT-PCR) clamping was performed for determination of EGFR mutation status. EGFR mutations were detected in five (35.7%) cell blocks of MPE, which showed results identical to those of the corresponding cell-free fluid, whereas mutations were detected in the plasma of only two (40.0%) of the five patients. Of seven patients treated with EGFR tyrosine kinase inhibitors (TKIs), EGFR mutations were detected in cell blocks, cell-free fluid of MPE, and plasma for only one of the four patients who responded to EGFR TKIs, while mutations were detected only in cell blocks of MPE and cell-free fluid of the three remaining patients. Our results suggest that detection of EGFR mutations in cell-free pleural fluid from lung adenocarcinoma patients using highly sensitive methods may be feasible, but that analysis of free plasma may lead to undetected mutations and misdiagnosis.

House dust mite (HDM) allergens have been associated with allergic diseases, such as asthma, allergic rhinitis, and atopic dermatitis. Various acaricidal agents have been suggested for control of house dust mites; however, their remains act as allergens even after death. Therefore, for avoidance of allergen, expelling the mites is a more effective policy than killing them. In this experiment, we compared the repellent effect of two essential oils (Matricaria chamomilla, Lavandula vera) against house dust mites, Dermatophagoids farinae and D. pteronyssinus in bed fabric. The essential oils were applied by direct contact method at various doses (0.1, 0.05, 0.025, 0.0125, and 0.00625 μl/cm2 and at various exposure times (30, 60, 120, 180, and 240 min). Results of this experiment suggest that the two oils have significant repellent activity. Camomile essential oil in 0.0125 μl/cm2 at 240 minutes had a repellent effect of 93.7% and lavender essential oil in 0.05 μl/cm2 at 180 minutes had a repellent effect of 88.9%. The results of this study showed that camomile essential oil has more potent repellent activity than lavender essential oil at a particular concentration.

This study demonstrated that hyaluronic acid (HA) accelerated peripheral nerve regeneration after crush injury to the common peroneal nerve in an experimental rabbit model. Ten male New Zealand White rabbits, weighing 1.8 to 2.0 kg, were used in this study. After creating the nerve crush model in every right leg, rabbits were divided into two groups. Animals in group A received application of HA into the area surrounding the crushed nerve, and group B was the sham control. Electrophysiological assessment was performed every week. After 10 weeks, nerve histological examination, muscle weight and muscle histology were used to evaluate regeneration of the injured common peroneal nerve. No differences in electrophysiological assessment were observed between the two groups. In peripheral nerve histology, myelinated nerve fibers were observed more frequently and less connective tissue was observed in the crushed nerve of group A. Fewer muscle degenerative changes, such as fibrosis, atrophy, and centrally located myonuclei, were detected in group A than in group B. In conclusion, HA could become a potential neuroprotective agent for improvement of peripheral nerve regeneration after crush injury.

Cytokines are known to function as regulatory molecules that can be produced by virtually every nucleated cell type in the body, including lymphocytes, monocytes/macrophages, epithelial cells, fibroblasts, and many others. Cytokines include lymphocyte-derived factors (lymphokines), monocyte-derived factors (monokines), hematopoietic factors (colony-stimulating factors), connective tissue/ growth factors, and chemotactic chemokines. Cytokines released in response to infection can affect tumor development in different ways. When exposed to infectious agents, cytokines are secreted by sentinel cells, such as macrophages and dendritic cells. These cytokines include interleukin 1 (IL-1) and tumor necrosis factor-α, as well as others, such as IL-6, IL-12, and IL-18. When released in sufficient quantities, these molecules can cause inflammation. Chronic inflammation is highly associated with tumor initiation, promotion, and progression. In this article, we review the roles and mechanisms of cytokines in tumor development.

Pneumatosis cystoides intestinalis (PCI) is a rare condition characterized by multiple intramural pockets of gas filled cysts in the intestinal wall. PCI is usually found incidentally on an imaging study. Many different causes of PCI have been suggested, including mechanical, pulmonary, and bacterial causes. Treatment is usually conservative, including oxygen and antibiotic therapy. We report on two cases of PCI, without symptoms, in a 62-year-old male and a 72-year-old male. Computed tomography showed numerous, small, round, and air densities on the sigmoid colon. Colonoscopy showed numerous, variable-sized, sessile polypoid, balloon-like distended, and protruding subepithelial masses covered with normal colonic mucosa on the sigmoid colon. We observed that when the cyst was stuck with a needle, the size of the cyst was reduced and showed a flat termination. Therefore, we made a diagnosis of PCI and report on the case with references.

Endocrine disrupting chemicals (EDCs) have detrimental effects on human health. Among these EDCs, bisphenol A (BPA) binds to estrogen receptors (ERs) to stimulate estrogen-mediated responses. BPA is assumed to disrupt the reproductive and developmental system of humans. In addition, BPA has recently been suspected as a risk of carcinogenesis. Because BPA can cause abnormal estrogen-mediated response in the organism, exposure to BPA may stimulate growth of estrogen-dependent breast cancers in human. In breast cancer, cyclin E and cyclin-dependent kinase inhibitor p27 are important in G1/S phase transition during cell cycle progression. In this study, using an MTT assay, we investigated the effect of BPA on proliferation of MCF-7 breast cancer cells in vitro. In addition, we also analyzed the transcriptional levels of cyclin E and p27 following treatment with BPA using semi-quantitative RT-PCR. As a result, treatment with BPA resulted in significant induction of breast cancer cell growth, compared to a vehicle. BPA caused alterations of cyclin E and p27 mRNA expression. Expression of cyclin E was increased by BPA, while p27 was decreased at 24 h after treatment with BPA in MCF-7 breast cancer cells. Taken together, these collective results suggest that exposure to BPA induced breast cancer cell proliferation with deregulation of the cell cycle. A further study is required in order to determine the effects of BPA on the carcinogenic process in in vivo models.

This study was conducted for evaluation of the alleviating effects of Phellodendrin cortex water extract (PCWE) on skin inflammation in hairless mice. Skin inflammation was induced by UVB irradiation and application of squalene monohydroperoxide (Sq-OOH) to the back skin of hairless mice for six weeks. At the same time, saline (C), jojoba oil (VC), PCWE (E), and 0.01% retinoic acid diluted with polyethylene glycol (PC) were applied topically twice per day, six days per week for a period of six weeks. The skin erythema index of the E group was lower than that of the C group. Epidermis and dermis of the C group were remarkably thickened, compared to the PC or E group. 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; the degree of degranulation was remarkable. However, these phenomena were alleviated in the PC and E2 groups. The E group showed a lower activity in skin xanthine oxidase but a higher activity in skin superoxide dismutase, compared to the C group (P<0.05). The VC, PC, and E groups also showed a high activity of skin catalase by 25.3%, 58%, and 42%, respectively, compared to the C group. Taken together, these results indicate that PCWE could have an alleviating efficacy on skin inflammation induced by UVB irradiation and application of Sq-OOH in hairless mice.

As a sensor of cellular energy status, AMP-activated protein kinase (AMPK) plays an important role in the pathophysiology of diabetes and its complications. Because AMPK is also expressed in podocytes, podocyte AMPK would be an important factor contributing to development of podocyte injury. We investigated the roles of AMPK in the pathological changes of podocyte synaptopodin induced by angiotensin II (Ang II), a major injury inducer. Mouse podocytes were incubated in media containing various concentrations of Ang II and AMPK-modulating agents, and the changes of synaptopodin were analyzed by confocal imaging and Western blotting. Ang II and compound C, an AMPK inhibitor, concentrated and re-localized synaptopodin from peripheral cytoplasm to the internal cytoplasm portion in podocytes. Ang II also reduced synaptopodin protein and mRNA, which were reversed by metformin and 5-aminoimidazole-4-carboxamide ribonucleoside. Losartan, an Ang II type 1 receptor antagonist, also recovered synaptopodin mRNA, which was suppressed by Ang II. We suggest that Ang II induces the relocation and suppression of podocyte synaptopodin by suppression of AMPK and via Ang II type 1 receptor, which would be an important mechanism in Ang II-induced podocyte phenotypical changes.

The objective of this study was to determine the effect of macrophages on growth of human colon cancer cells. The results showed that co-culture of colon cancer cells with macrophages inhibited the growth of colon cancer cells (HCT116 and SW620) depending on the number of macrophages, RAW 264.7 cells, and activated THP-1 cells accompanied by down regulation of pSTAT3 in cancer cells. We also found that expression and release of cancer cell growth inhibitory cytokines, IL-1 receptor antagonist (IL-1ra) and IL-10, was increased in macrophages. Blocking of the STAT3 pathway with specific inhibitor and siRNA of STAT3 abolished the growth of colon cancer cells and expression of IL-1ra and IL-10. In addition, neutralization of IL-1ra and IL-10 with antibodies resulted in reversal of macrophage-induced inhibition of cancer cell growth. These data showed that IL-1ra and IL-10 released from macrophages inhibit growth of colon cancer cells through inhibition of the STAT3 pathway.

Antimicrobial peptides are widely found in living organisms and are known to play a critical role in innate immunity. Numerous antimicrobial peptides from diverse species appear to be effective against pathogenic microorganisms of bacteria, fungi, protozoa, and viruses. Because antibiotic resistance is a global health issue in the fight against pathogenic microorganisms, there has been an urgent need for development of new antibiotic substances. In the current study, we performed yeast two hybrid screening using Beclin1 bait in order to find new peptide antibiotics from a random peptide library. Two candidate peptides from the screening were expressed in a yeast secretory system of Pichia pastoris and tested for any antimicrobial activity against Staphylococcus aureus, MRSA, MRSA2242, MRSA2250, Lactobacillus casei, and Lactobacillus acidophilus. Disc clear zone assay and spectrophotometric analysis revealed that the two peptides exert a decent activity against the pathogenic bacteria, in contrast to minimal effect on the commensal Lactobacillus strains. Taken together, this study presents novel peptides with antibacterial activity against the pathogenic forms of Staphylococcus aureus and suggests the possibility that these peptides, upon further characterization, may be developed as clinically useful antibiotics.

Macrophages can recognize antigens and microorganisms, and then initiate an appropriate defense. However, there has been a lack of comprehensive information regarding the genes that are modulated by commensal yeasts, including Saccharomyces cerevisiae or Saccharomyces exiguus. In addition, it is not clear to what extent the beneficial yeasts modulate the immune response against microbes and/or microbial toxins. Using DNA microarray, which contains approximately 25,000 genes, we studied interactions between host cells and yeast/bacterial toxin (LPS) by analyzing the transcriptional response of macrophages stimulated by Saccharomyces exiguus and/or Lipopolysaccharides. Thirty three genes were identified to be modulated by more than two folds between groups of macrophage cells. Pathway analysis provided insight into the mutual interactions. Of particular interest was the responses elicited by fungus in murine macrophage cells, including modulation of immunity/defense, cellular signal transduction, cell proliferation/differentiation, and transport. This finding indicates that the yeast induces immune response pathways as well as those associated with cell proliferation and transport. Among the 33 genes identified from the DNA microarray screening, eight genes were further checked by RT-PCR analysis using gene specific primers. Compared to those of negative control, sequential treatment with the yeast strain followed by LPS apparently induced expression of Tnfaip3, IL7R, and CD86, while it inhibited expression of Cxcl10 and CD83. In conclusion, this study identified the genes that are up-regulated by Saccharomyces exiguus. A further study is needed in order to determine whether these genes are modulated at the protein level, and also for their roles in control of immune responses.

Immunotherapeutic approaches using agonist antibodies or fusion proteins of immunomodulatory molecules significantly inhibit tumor growth and boost cell-mediated immunity. We isolated mRNA from previously reported 1D4 hybridoma cells and amplified the variable regions of the heavy chain (VH) and light chain (VL) genes using reverse-transcriptase polymerase chain reaction. Using a linker, the amplified sequences for the heavy and light chains were each connected to the sequence for a single polypeptide chain that was designed to be expressed. VL and VH fragments were cloned into the pOptiVEC-TOPO vector containing the human CH2-CH3 fragment. Then, 293T cells were transfected with the 1D4 single-chain Fv-Fc (scFv-Fc) constructs. A549 cells were used for presentation of the 1D4 antigen. Flow cytometry was performed for analysis of the secreted 1D4 scFv-Fc constructs. The DNA sequence of 1D4 scFv-Fc was obtained. The 1D4 scFv-Fc constructs were expressed by the transfected 293T cells and secreted into the culture medium. The immunoreactivity of the secreted scFv-Fc construct was lower than that of the murine 1D4 antibody for A549 cells. A 1D4 scFv-Fc construct for immunotherapy was developed.

In this study, a nanofibrous scaffold was obtained by co-electrospinning poly (3-hydroxybutyrate- co-3-hydroxyvalerate) (PHBV) and collagen in 2,2,2-trifluoroethanol at a ratio of 3/7. The fiber diameters were in the range of 250-600 nm. It was found that PHBV/Collagen (PHCP) nanofibrous scaffold showed greater proliferation than the PHBV nanofibrous scaffold induced by oxidant in NIH3T3 cells. Otherwise, in the early-stage wound-healing mouse model, wound closure was evaluated according to wound size reduction and histology of regenerated skin on the backs of mice. Each of the tissues removed on day 0, 3, 6, 9, 12, 15, and 18 was used for analysis of biochemical and pathological changes. None of the nanofiber-attached mice showed significant difference on the third day, however, from the third day until the ninth day, significantly faster healing was observed in PHCP-attached mice, compared to control wounds in epithelialization, wound contraction, and histopathological examinations. These results strongly support the beneficial effects of biomedical application of PHCP nanofiber in acceleration of the initial phase of wound healing through α-SM actin contraction.

The effects of β-lapachone on gastric secretion were investigated. The pylorus of male Sprague-Dawley rats was ligated and intraduodenally injected with β-lapachone, and the volume, pH, free HCl, and total acidity of gastric fluid were measured 6 hours after the operation. Treatment with β-lapachone resulted in dose-dependent inhibition of gastric secretion Gastric fluid was reduced to 42.9% of control level by 100 mg/kg of β-lapachone, leading to an increase of pH to 6.70 from 1.85 in the control group. In parallel with the increase of pH, at this dosage, free HCl and total acidity decreased to 16.7% and 12.0%, respectively, of control levels. β-Lapachone exhibited ED50 values of 72, 46, and 47 mg/kg for inhibition of gastric volume, free HCl, and total acidity, respectively, implying a superior efficacy on gastric acid to volume. In comparison, pantoprazole (30 mg/kg) reduced the volume, free HCl and total acidity of gastric fluid to 53.0%, 26.0%, and 25.0%, respectively, of control levels, resulting in an increase in pH to 6.36. In the current study, it was confirmed that β-lapachone at an appropriate dose (100 mg/kg) exerted a higher inhibitory effect on gastric secretion than pantoprazole (30 mg/kg), a well-known proton-pump inhibitor. Therefore, it is suggested that β-lapachone could be a candidate compound for prevention or treatment of gastric ulcers induced by diverse psychological and physical stimuli.

The current study was designed to investigate the effect of Red ginseng extract on development of colonic aberrant crypt foci (ACF) induced by 1,2-dimethylhydrazine (DMH) in male F344 rats. Five-week old animals received subcutaneous injections of DMH (30 mg/kg body weight) four times, for a period of two weeks in order to induce ACF. The animals were divided into groups fed a diet containing red ginseng extract at three different doses (0.5, 1.0, and 2.0%), respectively. Animals were evaluated for the total number of ACF and total aberrant crypts (AC) per colon detected from methylene blue-stained rat colon. ACF formation was observed in animals in the DMH-treated group. Four-time treatment with DMH induced mean 265.8±48.3 ACF/colon composed of a total of 608.8±110.9 aberrant crypts AC/colon. The numbers of ACF and AC induced by DMH were decreased to 204.4±29.3 and 464.7±70.3 by treatment with 0.5% red ginseng extract. In addition, the number of large ACF (≥4 AC/ACF) was suppressed from 39.9±10.6 ACF/colon in control to 28.5±5.3 ACF/colon in 0.5% red ginseng extract. These results suggested that red ginseng extract exerted a chemopreventive effect on DMH-induced colon cancer by inhibiting development of ACF and AC in F344 rats.