Standardized Extract of Atractylodis Rhizoma Alba and Fructus Schisandrae Ameliorates Coughing and Increases Expectoration of Phlegm.

Cough and phlegm frequently occur in respiratory diseases like upper respiratory tract infections, acute bronchitis, and chronic obstructive pulmonary diseases. To relieve these symptoms and diseases, various ingredients are being used despite the debates on their clinical efficacy. We aimed to investigate the effects of the extract CKD-497, composed of Atractylodis Rhizoma Alba and Fructus Schisandrae, in relieving cough and facilitating expectoration of phlegm. CKD-497 was found to inhibit inflammatory mediators such as interleukin-8 (IL-8) and tumor necrosis factor α (TNF-α) in lipopolysaccharide (LPS)-treated mouse macrophages and transient receptor potential cation channel 1 (TRPV-1)-overexpressed human bronchial epithelial cells stimulated by capsaicin. CKD-497 decreased the viscosity of the mucin solution. During in vivo experiments, CKD-497 reduced coughing numbers and increased expectoration of phlegm via mucociliary clearance enhancement. Collectively, these data suggest that CKD-497 possesses potential for cough and phlegm expectoration treatment.

Inhibitory effects of Saururi chinensis extracts on melanin biosynthesis in B16F10 melanoma cells.

Saururus chinensis has been used in folk medicine in Korea for the treatment of edema, jaundice, gonorrhea, and several inflammatory diseases. Saururi chinensis extracts (SCE) have demonstrated anti-inflammatory and anti-oxidant activities, as well as anti-asthmatic, antihypertensive, anti-angiogenic, and therapeutic activities for atopic dermatitis. However, the inhibitory activity of SCE on the melanogenesis signaling pathway is not completely understood. This study examined the effects of SCE on the melanogenesis signaling pathway activated by α-melanocyte-stimulating hormone (α-MSH). We found that SCE inhibited melanin production in a dose-dependent manner without causing cytotoxicity in B16F10 cells. Interestingly, SCE decreased α-MSH-induced tyrosinase activity in B16F10 cells but did not inhibit tyrosinase activity under cell-free conditions. The results of this study indicate that SCE may reduce pigmentation by way of an indirect, nonenzymatic mechanism. We also found that SCE decreased α-MSH-induced microphthalmia-associated transcription factor (MITF) and tyrosinase expression and induced the activation of extracellular signal-regulated kinase (ERK). These results suggest that the depigmenting effect of SCE may result from downregulation of MITF and tyrosinase expression due to increased ERK activity. Thus, our results provide evidence that SCE might be useful as a potential skin-whitening agent.

Fractional photothermolysis laser treatment of male pattern hair loss.

BACKGROUND: Various trials have been conducted on the management of male pattern hair loss (MPHL). A variety of laser and light sources have been used for the treatment of MPHL. OBJECTIVE: To understand the effects of a 1,550-nm fractional erbium-glass laser on the hair cycle in an alopecia mouse model and to study the clinical effects of the same laser used as treatment for MPHL. MATERIALS AND METHODS: Irradiation was applied to the shaved skin of C3H/HeN mice using various energy and density settings and varied irradiation intervals. In a clinical pilot study involving human subjects, 20 participants were treated over five sessions at 2-week intervals. A fractional photothermolysis laser was used at the energy of 5 mJ and a total density of 300 spots/cm(2). RESULTS: In the animal study, the hair stimulation effects were dependent upon the energy level, density, and irradiation interval. The anagen conversion of hair and the increase in Wnt 5a, ß-catenin signals were observed. In the human pilot study, incremental improvements in hair density and growth rate were observed. CONCLUSIONS: This pilot study showed that a 1,550-nm fractional erbium-glass laser might induce hair growth, but more intensive studies are required to clarify the clinical applications of this treatment.

The safety and efficacy of CKD-497 in patients with acute upper respiratory tract infection and bronchitis symptoms: a multicenter, double-blind, double-dummy, randomized, controlled, phase II clinical trial.

BACKGROUND: Acute upper respiratory tract infection (AURI) together with acute bronchitis is the most common illness worldwide. Botanical medicines used as expectorants and antitussives have proven to be effective while also having excellent safety margins. We aimed at evaluating the efficacy and safety of a new botanical drug, CKD-497, in patients with AURI and acute bronchitis. METHODS: In this phase 2 study, 225 patients were enrolled and randomly assigned to one of four treatment groups: placebo (n=55), Synatura® (n=49), CKD-497 200 mg (n=68), or CKD-497 300 mg (n=53). The study drugs were administered three times daily over the course of 7 days. Primary endpoint was the change in the bronchitis severity score (BSS) from baseline to day 7. Secondary endpoint was evaluated based on clinical response rates on days 4 and 7. A safety analysis was also performed. RESULTS: Between baseline and day 7, the mean BSS scores decreased significantly in each group (P<0.001): -4.04±1.85, -4.31±1.47, -4.09±1.48, and -4.28±1.69. However, neither the CKD-497 nor Synatura® group showed any significant effect on the difference in BSS change (P=0.75). The rate of clinical response was higher in the CKD-497 300 mg group as compared to the placebo only on day 4 (36% vs. 18%; P<0.05) and those having more severe bronchitis (phlegm score ≥3) showed a significant reduction of total BSS in the Synatura® and CKD-497 groups (P=0.042). No significant adverse events were observed in either of the CKD-497 groups. CONCLUSIONS: CKD-497 and even the positive control drug had no significant effect on BSS change in this phase 2 clinical trial. However, CKD-497 300 mg had a mild but significant clinical improvement in early bronchitis patients with more severe phlegm. Considering both efficacy and safety, a future study using 300 mg of CKD-497 with a shorter-term endpoint is warranted in patients with more severe bronchitis symptoms.

Leucine-rich glioma inactivated 3 induces neurite outgrowth through Akt and focal adhesion kinase.

Leucine-rich glioma inactivated 3 (LGI3) is a secreted protein that belongs to LGI/epitempin family. LGI3 is highly expressed in brain in a transcriptionally and developmentally regulated manner. Here we found that LGI3 induced neurite outgrowth in Neuro-2a cells and dorsal root ganglia explants. LGI3 treatment or overexpression increased neurite outgrowth and knockdown of LGI3 by siRNA had opposite effect. LGI3 treatment increased phosphorylation of Akt and a 125-kDa protein. Immunoprecipitation identified the 125-kDa protein as focal adhesion kinase (FAK). LGI3 overexpression increased phospho-Akt, phospho-FAK and FAK protein. Inhibition of Akt activation by PI3 kinase inhibitor attenuated LGI3-induced FAK phosphorylation and neurite outgrowth. Taken together, we propose that LGI3 is a neuritogenic factor whose signaling pathway involves Akt-mediated FAK activation.

Clinicopathologic efficacy of copper bromide plus/yellow laser (578 nm with 511 nm) for treatment of melasma in Asian patients.

BACKGROUND: Melasma is a common pigmentary disorder in Asians. Although the pathogenesis of melasma is not yet fully understood, there are several hypotheses supporting angiogenetic factors related to some types of melasma. OBJECTIVE: To test the efficacy of copper bromide laser in the treatment of Korean women with melasma. MATERIALS AND METHODS: Clinical parameters included physician and patient assessment and Melasma Area and Severity Index score. The intensity of pigmentation and erythema was measured using a chromometer. To evaluate histopathologic changes, punch biopsies from melasma were obtained from four patients. Immunohistochemical staining for Melan-A, endothelin 1, CD34, and vascular endothelial growth factor (VEGF) antigen of the melasma lesions was observed. RESULTS: Mean MASI score decreased dramatically after treatment. Patients exhibited telangiectatic erythema within the melasma lesion. The values of L(*) reflecting intensity of pigmentation increased, and the values of a(*) as the measurement of redness decreased after the treatments. Expression of Melan-A, CD34, endothelin-1, and VEGF decreased after treatment. CONCLUSION: The potential application of an antiangiogenetic laser for the treatment of melasma specially accompanied by pronounced telangiectasia in Asian skin is a possible treatment option.

Synergistic Inhibition of Tumor Necrosis Factor-Alpha-Stimulated Pro-Inflammatory Cytokine Expression in HaCaT Cells by a Combination of Rapamycin and Mycophenolic Acid.

BACKGROUND: Keratinocytes release various pro-inflammatory cytokines, chemokines, and adhesion molecules such as intercellular adhesion molecule 1 (ICAM-1) in response to cytokines such as tumor necrosis factor (TNF)-α and interferon (IFN)-γ. Rapamycin and mycophenolic acid (MPA) have potent immunosuppressive activity because they inhibit lymphocyte proliferation. OBJECTIVE: We investigated the effects of rapamycin and MPA on the expression of inflammation-related factors such as ICAM-1 and inducible nitric oxide synthase (iNOS), pro-inflammatory cytokines and chemokines, and related signaling pathways in TNF-α-stimulated HaCaT cells. METHODS: The viability of HaCaT cells treated with rapamycin and MPA was confirmed using MTT assay. The expression of various cytokines such as interleukin (IL)-1ß, IL-6, and IL-8; inflammation-related factors such as ICAM-1 and iNOS; and the activation of mitogen activated protein kinase (MAPK) signaling pathways mediated by extracellular signal-related kinases (ERK), p38, and c-Jun N-terminal kinases (JNK) in TNF-α-stimulated HaCaT cells were confirmed using reverse transcription-polymerase chain reaction and western blotting. RESULTS: Combined treatment of TNF-α-induced HaCaT cells with rapamycin and MPA decreased ICAM-1 and iNOS expression and ERK and p38 activation more than treatment with either drug alone. The most significant decrease was observed with a combination of rapamycin (80 nM) and MPA (20 nM). These results show that co-treatment with these agents has a synergistic anti-inflammatory effect by blocking the activation of the ERK/p38 MAPK signaling pathway and thus suppressing the TNF-α-induced expression of ICAM-1 and iNOS. CONCLUSION: The combination of rapamycin and MPA could potentially be used as a therapeutic approach in inflammatory skin diseases.

Berberine regulates melanin synthesis by activating PI3K/AKT, ERK and GSK3ß in B16F10 melanoma cells.

Berberine, an isoquinoline alkaloid, has a wide range of beneficial properties, including anti-bacterial, anti-inflammatory, anti-cancer, and cholesterol-lowering effects. Recently findings suggest that berberine improves glucose and lipid metabolism disorders. In the present study, we examined the mechanism underlying the inhibitory effect of berberine on α-melanocyte-stimulating hormone (α-MSH)-stimulated B16F10 melanoma cells. The results showed that berberine attenuated α-MSH induction of the microphthalmia-associated transcription factor (MITF) and tyrosinase in a dose-dependent manner. To elucidate the mechanism underlying the inhibitory effect of berberine, we examined the effect of α-MSH-stimulated phosphorylation of PI3K/AKT, ERK, and GSK3ß. The results showed that treatment with berberine resulted in a reduction in the phosphorylation of PI3K/AKT, ERK, and GSK3ß. Taken together, the results suggested that berberine inhibits melanin synthesis and tyrosinase activity by downregulating the expression of MITF and tyrosinase. Thus, these findings may contribute to the potential application of berberine in the prevention and treatment of skin pigmentation disorders.

Safety evaluation of stamp type digital microneedle devices in hairless mice.

BACKGROUND: Microneedles provide a minimally invasive means to transport molecules into the skin. A number of specific strategies have been employed to use microneedles for transdermal delivery. OBJECTIVE: The purpose of this study was to investigate the safety of two new digital microneedle devices (Digital Hand® and Digital Pro®; Bomtech Electronics Co., Ltd., Seoul, Korea) for the perforation of skin in skin-hairless-1 mice. This device replaces conventional needles and is designed specifically for intradermal delivery. METHODS: We used two newly developed digital microneedle devices to perforate the skin of skin-hairless-1 mice. We conducted a comparative study of the two digital microneedle devices and DTS® (Disk type-microneedle Therapy System; DTS lab., Seoul, Korea). To evaluate skin stability, we performed visual and dermatoscopic inspections, measurements of transepidermal water loss, and biopsies. RESULTS: The two novel digital microneedle devices did not induce significant abnormalities of the skin on visual or dermatoscopic inspection, regardless of needle size (0.25~2.0 mm). No significant histopathological changes, such as inflammatory cell infiltration, desquamation of the stratum corneum, or disruption of the basal layer, were observed. The digital microneedle devices and microneedle therapy system produced similar results on measures of skin stability. CONCLUSION: These two novel digital microneedle devices are safe transdermal drug delivery systems.

A comparison of neuropeptide expression in skin with allergic contact dermatitis in human and mouse.

BACKGROUND: Allergic contact dermatitis (ACD) is a manifestation of a cell-mediated immune response, but its mechanism remains unknown. Recently, we investigated whether ACD involves various neuropeptides. Substance P (SP) is a neuropeptide that is known to act as a neurokinin receptor when the immune response is initiated. Calcitonin gene-related peptide (CGRP) is a distinct typical neuropeptide and, with SP, induces the immune response. Neuropeptides in neurogenic inflammation are regulated by the inactivation of receptors and enzymes that can cause neuropeptide degradation (e.g. angiotensin-converting enzyme [ACE]), but no enzyme that can degrade SP and CGRP has yet been reported. METHODS: We investigated changes in the expression of SP and CGRP, as representative of typical neuropeptides, in ACD skin in mouse and human and the effect of ACE expression on the degradation of these neuropeptides using reverse transcription polymerase chain reaction and immunoblot assay. We also examined the relationship between ACD and neuropeptides in skin tissue from human ACD subjects and mice with induced ACD by analyzing cytokine expression and the results of hematoxylin and eosin staining and immunofluorescence assay. RESULTS: Expression of SP, CGRP, and ACE was higher in skin tissues from animals with acute ACD than in normal animal skin. However, CGRP expression in human skin with acute ACD was lower than in normal skin, unlike expression of SP and ACE, both of which were higher in ACD human skin than in normal human skin. CONCLUSIONS: We found different patterns of neuropeptide expression in human versus mouse skin. These neuropeptide activities were influenced by an increase in neuropeptide degrading enzymes. Our findings show that when SP is produced, expression of CGRP is suppressed in human skin with ACD. The reduction of CGRP expression in patients with acute ACD is caused by mast cells activated by SP.

Transdermal drug delivery using disk microneedle rollers in a hairless rat model.

BACKGROUND: Transdermal drug delivery systems (TDDSs) represent more reliable and consistent methods of drug dosing than oral administration. However, TDDSs can administer only low molecular weight (MW) drugs and require a power source. Disk microneedle rollers facilitate the passage of low and high MW substances through the direct perforation of the stratum corneum and dermis, without stimulating dermal nerves. OBJECTIVES: We investigated in vitro whether disk microneedle rollers, developed for the Diskneedle Therapy System (DTS™) in South Korea, can deliver drugs effectively through the skin of hairless rats. METHODS: The disk microneedle rollers used in the DTS™ are metal and consist of several plates bearing microneedles of graded lengths (0.15 mm, 0.25 mm, 0.50 mm). To test in vitro permeation, the skin of a hairless rat was mounted in a Franz diffusion cell system and rolled with a disk roller without microneedles and with rollers fitted with microneedles of each size. Rhodamine B base (80 µl) was applied to the skin for 24 hours, 48 hours, and 72 hours, and dye permeation was detected at 543 nm. Dye binding to the skin was also confirmed using fluorescence microscopy at six hours after the application of rhodamine B. RESULTS: Use of the disk microneedle roller increased the skin penetrance of rhodamine B base in hairless rats in accordance with microneedle length, as assessed using a fluorescence penetration test. CONCLUSIONS: Disk microneedle rollers, as designed for the DTS™, can be used for transdermal drug delivery. Microneedles can be selected according to the length appropriate for each application.

Inhibitory Effect of Vitamin U (S-Methylmethionine Sulfonium Chloride) on Differentiation in 3T3-L1 Pre-adipocyte Cell Lines.

BACKGROUND: S-methylmethionine sulfonium chloride was originally called vitamin U because of its inhibition of ulceration in the digestive system. Vitamin U is ubiquitously expressed in the tissues of flowering plants, and while there have been reports on its hypolipidemic effect, its precise function remains unknown. OBJECTIVE: This study was designed to evaluate the anti-obesity effect of vitamin U in 3T3-L1 pre-adipocyte cell lines. METHODS: We cultured the pre-adipocyte cell line 3T3L1 to overconfluency and then added fat differentiation-inducing media (dexamethasone, IBMX [isobutylmethylxanthine], insulin, indomethacin) and different concentrations (10, 50, 70, 90, 100 mM) of vitamin U. Then, we evaluated changes in the levels of triglycerides (TGs), glycerol-3-phosphate dehydrogenase (G3PDH), AMP-activated protein kinase (AMPK), adipocyte-specific markers (peroxisome proliferator-activated receptor γ [PPAR-γ], CCAAT/enhancer-binding protein α [C/EBP-α], adipocyte differentiation and determination factor 1 [ADD-1], adipsin, fatty acid synthase, lipoprotein lipase) and apoptosis-related signals (Bcl-2, Bax). RESULTS: There was a gradual decrease in the level of TGs, C/EBP-α, PPAR-γ, adipsin, ADD-1 and GPDH activity with increasing concentrations of vitamin U. In contrast, we observed a significant increase in AMPK activity with increasing levels of vitamin U. The decrease in bcl-2 and increase in Bax observed with increasing concentrations of vitamin U in the media were not statistically significant. CONCLUSION: This study suggests that vitamin U inhibits adipocyte differentiation via down-regulation of adipogenic factors and up-regulation of AMPK activity.

The Safety Evaluation of a Potent Angiogenic Activator, Synthetic Peptide (SFKLRY-NH2) for the Skin Application.

A novel synthetic hexapeptide (SFKLRY-NH2) that displays angiogenic activity has been identified by positional scanning of a synthetic peptide combinatorial library (PS-SPCL). This study was carried out to investigate the irritation of the SFKLRY-NH2 on the skin. The tests were performed on the basis of Korea Food and Drug Administration (KFDA) guidelines. In results, cell toxicity is not appeared for SFKLRY-NH2 in HaCaT cells and B16F10 cells. SFKLRY-NH2 induced no skin irritation at low concentration (10 µM), mild irritation at high concentration (10mM). We consider that this result is helpful for saying about the safety of SFKLRY-NH2 in clinical use.

Successful treatment of alopecia areata with topical calcipotriol.

Alopecia areata (AA) is an inflammatory hair loss of unknown etiology. AA is chronic and relapsing, and no effective cure or preventive treatment has been established. Vitamin D was recently reported to be important in cutaneous immune modulation as well as calcium regulation and bone metabolism. It is well known that areata is common clinical finding in patients with vitamin D deficiency, vitamin D-resistant rickets, or vitamin D receptor (VDR) mutation. The biological actions of vitamin D3 derivatives include regulation of epidermal cell proliferation and differentiation and modulation of cytokine production. These effects might explain the efficacy of vitamin D3 derivatives for treating AA. In this study, we report a 7-year-old boy with reduced VDR expression in AA, recovery of whom was observed by topical application of calcipotriol, a strong vitamin D analog.

Distribution of malassezia species on the scalp in korean seborrheic dermatitis patients.

BACKGROUND: Malassezia species play an important role in the pathogenesis of seborrheic dermatitis. In particular, M. restricta and M. globosa are considered to be the predominant organisms in seborrheic dermatitis of Western countries. However, species distribution of Malassezia in seborrheic dermatitis has not been clearly determined yet in Asia. OBJECTIVE: To identify the distribution of Malassezia species on the scalp of seborrheic dermatitis patients in Korea using 26S rDNA PCR-RFLP analysis. METHODS: A total of 40 seborrheic dermatitis patients and 100 normal healthy volunteers were included in this study. For the identification of Malassezia species, the scalp scales of the subjects were analyzed by 26S rDNA PCR-RFLP analysis. RESULTS: The most commonly identified Malassezia species were M. restricta in the seborrheic dermatitis patients, and M. globosa in the normal controls. In the seborrheic dermatitis group, M. restricta was identified in 47.5%, M. globosa in 27.5%, M. furfur in 7.5%, and M. sympodialis in 2.5% of patients. In the healthy control group, M. globosa was identified in 32.0%, M. restricta in 25.0%, M. furfur in 8.0%, M. obtusa in 6.0%, M. slooffiae in 6.0%, and M. sympodialis in 4.0% of subjects. CONCLUSION: M. restricta is considered to be the most important Malassezia species in Korean seborrheic dermatitis patients.

Effects of vitamin C vs. multivitamin on melanogenesis: comparative study in vitro and in vivo.

BACKGROUND: Vitamin C has been used for the treatment of hyperpigmented diseases. However, there is no study available on hypopigmenting effect of multivitamin. OBJECTIVES: To investigate the inhibitory effects of multivitamin and vitamin C on melanogenesis. METHODS: We assessed the effect of multivitamin and vitamin C on cell viability, melanogenesis, and mushroom tyrosinase. The antioxidant activity of multivitamin and vitamin C was measured. We performed the Western blot analysis to study the effect of multivitamin and vitamin C on the expression of tyrosinase, microphthalmia-associated transcription factor (MITF), extracellular signal-regulated kinase (ERK), and Akt/protein kinase B. In a clinical trial, 20 melasma patients were treated with split face iontophoresis using either multivitamin or vitamin C. We evaluated the hypopigmenting effects of multivitamin and vitamin C through colorimetric measurement. RESULTS: Both vitamin C and multivitamin inhibited melanogenesis with low cytotoxicity. Multivitamin reduced melanin contents greater than vitamin C. However, the effects of vitamin C are greater than those of multivitamin on mushroom tyrosinase inhibition and antioxidation. In the Western blot, the reduced tyrosinase expression and MITF level were observed only in multivitamin-treated group, and not in vitamin C-treated group. No changes of ERK and Akt activation were observed in both multivitamin and vitamin C-treated groups. After 12 weeks of treatment with iontophoresis, both multivitamin and vitamin C were effective for melasma. CONCLUSIONS: Multivitamin has shown more anti-melanogenic effect than vitamin C via the downregulation of MITF.