Ionizing radiation has long been used in medicine since the discovery of X-rays. Diagnostic imaging using synchrotron radiation has been under investigation since Rubenstein et al. reported dual-energy iodine-K-edge subtraction coronary angiography. Recently, computed tomography (CT) and magnetic resonance imaging (MRI) have provided better quality results than conventional radiology, providing important information on human internal structures. However, such techniques are unable to detect fine micron sized structures for the early diagnosis of tumors, vascular diseases and other medical objectives. Third generation synchrotron X-rays are well known for their superiority in coherence and energy tunability with respect to conventional X-rays. Consequently, new contrast mechanisms with a superior spatial resolution are becoming available. Here we present the extremely fine details of live animal internal structures using unmonochromatized synchrotron X-rays (white beam) and a simple detector system. Natural movements of the internal organs are also shown. The results indicate that this imaging technique can be applied to investigating microstructures and evaluating the function of the internal organs. Furthermore, this imaging system may be applied to humans as the next tool beyond CT and MRI.
Animal ; *Diagnostic Imaging ; Male ; Mice ; Mice, Inbred HRS ; *Synchrotrons

Recently, a topical preparation of 5% propantheline bromide and 10% aluminum hydroxychloride in an emulsion ground substance was introduced as an effective antiperspirant. According to the literature, aluminum salts may produce functional closure of swet ducts. Nevertheless, histologic study has as yet failed to reveal a solid anatomic basis for occlusion. In this study, the scanning electron microscope (SEM) was used to demonstrate eccrine gland ostial occlusion due to topical 5% propantheline bromide and 10% aluminum hydroxychloride.
Aluminum Hydroxide/pharmacology* ; Animal ; Eccrine Glands/drug effects* ; Foot ; Male ; Mice ; Mice, Inbred HRS ; Microscopy, Electron, Scanning ; Propantheline/pharmacology* ; Sweat Glands/drug effects*

Equol, an isoflavonoid metabolite produced from the dietary isoflavone daidzein by the gut microflora in mammals, has been found to protect not only against ultraviolet (UV) radiation-induced cutaneous inflammation and photoimmune suppression, but also have antiphotocarcinogenic properties in mice. Because the state of DNA damage has been correlated with suppression of the immune system and photocarcinogenesis, we have therefore examined the potential of equol to offer protection from solar-simulated UV (SSUV) radiation-induced DNA damage in hairless mice by the immunohistochemical approach using monoclonal antibody specific for cyclobutane pyrimidine dimers (CPDs; H3 antibody). Topical application of 20 micrometer equol lotion, which was applied both before and after SSUV significantly reduced the number of CPDs. This reduction was evident immediately after SSUV exposure, at 1 h after exposure, and at 24 h after exposure, revealing 54%, 50%, and 26% reduction in CPDs, respectively. When the same concentration was applied for 5 consecutive days after SSUV exposure, there was no significant difference in the reduction of CPDs immediately after SSUV irradiation or at 1 hour afterwards, but there were significant reductions of 23% and 42% at 24 and 48 h after SSUV exposure, respectively. Despite apparently reducing the number of CPDs post-SSUV, topically applied equol did not appear to increase the rate of dimer removal. To conclude, equol applied topically prior to SSUV irradiation offers protection against CPD formation in hairless mice, possibly by acting as a suncreen and thus inhibiting DNA photodamage.
Administration, Topical ; Animals ; DNA/drug effects/radiation effects ; *DNA Damage ; Female ; Immunohistochemistry ; Isoflavones/*pharmacology ; Mice ; Mice, Inbred HRS ; Pyrimidine Dimers/metabolism ; Skin/drug effects/metabolism/*radiation effects ; Sunlight/adverse effects ; Ultraviolet Rays/*adverse effects

BACKGROUND: Stratum corneum lipids serve as a water retainer as well as permeability barrier by forming a multi-lamellae structure in the stratum corneum. The major constituent of these lipids, ceramides, have been shown to be predominantly associated with both functions. OBJECTIVE: Exposure of human epidermis to ultraviolet(UV) irradiation leads to changes in the physiologic and biochemical features of the skin. In order to investigate the effect of UVB irradiation on the skin barrier function in hairless mice, transepidermal water loss (TEWL) and lipid composition of stratum corneum were evaluated in hairless mice. METHODS: Hairless mice were irradiated 3 times weekly for 3 weeks with suberythemal dose (0.6MED, Group I) and minimal erythemal dose(MED), Group II) of UB. The mice of Group III received high dose of UVB(3MED) on the back in a single exposure. The control was Group IV. TEWL measured by evaporimeter and lipid composition of stratum corneum appraised by high performance thin layer chromatography(HPTLC) were evaluated weekly for 3 weeks. RESULTS: 1. Each time it was measured, the values of TEWL in group I were lower than group IV, but the difference was not significant. The peak value of TEWL in group II was 8.2+/-1.56 g/cm/h on the 7th day. The increase in TEWL was markedly significant at this point(P<0.001). Although the values of TEWL on the 14th and 21th day in group E increased compared with those of the control group, the significance of the values decreased (P<0.05). 2. The peak value of TEWL in group III was 9.88+/-1.13 g/cm/h on the 2nd day, showed a markedly significant increase compared with that of the control group(P<0.001). The values of TEWL decreased to the level of the control group on the 14th day. 3. The lipid(cholesterol sulfate, ceramide and neutral lipid) and total lipid mass in group 1 were insignificantly larger than that of the group IV measured each time. On the 7th and 14th day, the amount of each three lipid and total lipid mass significantly increased (P<0.05). On the 21th day, the amount of ceramide and neutral lipid showed a significant increase(P<0.05), furthermore the total lipid mass increased pronouncedly(P<0.01) in group II. 4. The amount of the 3 kinds of lipid and total lipid mass in group III significantly increased compared with those of the control group on the 2nd day(P<0.05). After the 7th day, no significant difference of the lipid mass except neutral lipid compared with that of the control group was seen. Comparing the 2nd and 14th day, there was a significant decrease in the amount of ceramide and total lipid mass(P<0.05) CONCLUSION: These results results suggest that considerable amount of UVB irradiation given in single or repeated exposure causes the disruption of skin barrier function, but a compensatory increase of skin lipid, especially ceramide, protests it from further damage and also improves skin barrier function.
Animals ; Ceramides ; Epidermis ; Humans ; Mice ; Mice, Hairless* ; Permeability ; Skin*

Chinese herbal medicine ultrafine powder has become a research hotspot for the addition of cosmetic raw materials. Dendrobium candidum is a traditional Chinese herbal medicine. Its extract and stem extract are already cosmetic raw materials and its water extract has the effect of preventing photoaging,but D. candidum ultrafine powder has not been accepted as a raw material for cosmetics,and no relevant research on photoaging prevention has been reported. In this experiment,the ultra-fine powder and fine powder of D. candidum to prevent photoaging were observed and compared,and its mechanism of action was discussed to provide a basis for the prevention of skin photoaging products. Seventy-two female ICR mice were randomly divided into normal group,model group,solvent group,titanium dioxide(Ti O2) group,isooctyl salicylate(2-ES) group,D. candidum ultrafine powder 1(DP1),ultrafine powder 2(DP2) and fine powder(DP3) groups. The photoaging model was established by ultraviolet irradiation for 8 weeks,and the model was intervened while modeling. The skin wrinkle grade,elastic parameters,skin microcirculation blood flow,skin structure and pathological changes(skin thickness,skin collagen fiber,elastic fiber) were observed,the skin transforming growth factor-β1(TGF-β1),Smad3 levels were determined,and the type Ⅰ and type Ⅲ collagen,matrix metalloproteinase-1(MMP-1),activated protein-1(AP-1),VEGF expression were detected. The results showed that ultrafine powder(DP1,DP2) significantly reduced the wrinkle level and skin blood flow of the model mice(P<0. 05,P<0. 01); DP1,DP2 and DP3 could significantly reduce the thickness of the epidermis(P<0. 001),improve collagen fiber,elastic fiber hyperplasia,and distortion and decrease VEGF expression,and DP1 is better than DP2 and DP3; each group could up-regulate type Ⅰ collagen,down-regulate type Ⅲ collagen,AP-1,MMP-1 protein expression,and DP1 improvement optimal. However,it has no obvious effect on TGF-β1 and Smad3. The ultrafine powder and fine powder of D. candidum have certain preventive effect on photoaging,and the effect of ultrafine powder is better than that of fine powder. Ultrafine powder may down-regulate the expression of type Ⅲ collagen,AP-1 and MMP-1 by up-regulating type Ⅰ collagen. Inhibition of collagen degradation plays a role in preventing photoaging.
Animals ; Dendrobium ; Female ; Mice ; Mice, Hairless ; Mice, Inbred ICR ; Skin ; Skin Aging ; Ultraviolet Rays

S-methyl-(L)-methionine (SMM), also known as vitamin U, is commercially available as skin care cosmetic products for its wound healing and photoprotective effects. However, the low skin permeation expected of SMM due to its hydrophilic nature with a log P value of −3.3, has not been thoroughly addressed. The purpose of this study thus was to evaluate the effect of skin permeation enhancers on the skin permeation/deposition of SMM. Among the enhancers tested for the in vitro skin permeation and deposition of SMM, oleic acid showed the most significant enhancing effect. Moreover, the combination of oleic acid and ethanol further enhanced in vitro permeation and deposition of SMM through hairless mouse skin. Furthermore, the combination of oleic acid and ethanol significantly increased the in vivo deposition of SMM in the epidermis/dermis for 12 hr, which was high enough to exert a therapeutic effect. Therefore, based on the in vitro and in vivo studies, the combination of oleic acid and ethanol was shown to be effective in improving the topical skin delivery of SMM, which may be applied in the cosmetic production process for SMM.
Animals ; Ethanol ; In Vitro Techniques* ; Mice ; Mice, Hairless ; Oleic Acid ; Skin Care ; Skin* ; Vitamin U ; Wound Healing

BACKGROUND: Acetone disrupts the cutaneous permeability barrier through the removal of stratum corneum lipids. This pertubation of barrier integrity stimulates a variety of homeostatic repair that ultimately results in the normalization of barrier function. OBJECT: To measure the effect of steroid on the barrier recovery of acetone applied skin. MATERIAL AND METHODS: The flank skin of 8~10 week old hairless mice was treated with acetone and then topical and systemic steroids were applied. Transepidermal water loss(TEWL) was checked after 0, 3, 6, 12 and 24 hours. Electron and light microscopic examination and ion capture cytochemistry were performed after 3, 6, 12 and 24 hours after systemic and topical steroids had been applied. RESULTS: The results were as follows ; 1) During 3~6 hours after experiment, the recovery rate of TEWL was most prominent in the group of acetone applied animal than other groups. 2) After 12 hours after acetone applied, formation of new stratum corneum was found in the groups of acetone applied or acetone applied skin with topical steroid application. But loss of stratum corneum was observed in the groups of high or low dose steroid injection. 3) Ruthenium tetroxide staining of acetone alone or topical steroid treated specimens after 12 hours experiment revealed that the lipid bilayer was partly impaired and fragmented. Intercellular spaces were widening and the lipid bilayer disappeared or was damaged in the groups of high or low dose steroid injection. 4) Six hours after acetone application, pattern of calcium distribution had been partially reestabilished in the group of acetone alone or topical steroid treated animals. But calcium content was still sparse and decreased from the stratum granulosum to basale in the groups of high or low dose steroid injection. CONCLUSION: In summary the present study demonstrates that steroid treatment acutely delays recovery rate of TEWL, inhibits normalization of calcium gradient or epidermal lipid synthesis that leads to abnormalities in permeability barrier homeostasis.
Acetone ; Animals ; Calcium ; Extracellular Space ; Histocytochemistry ; Homeostasis ; Lipid Bilayers ; Mice ; Mice, Hairless* ; Permeability ; Ruthenium ; Skin ; Steroids

BACKGROUND: Topical irritants disrupt the cutaneous permeability barrier through the removal of stratum comeum lipids. This perturbation of barrier integrity stimulates a variety of homeostatic repair responses that ultimately result in the normalization of bamer function. Object To measure the effect of desoxymethasone ointment, vaseline and hydrobase on the barrier recovery of benzalkonium chloride (BKC) imtated skin. MATERIALS AND METHODS: The left flank skin of 2-3 monthold hairless mice was treated with BKC and then desoxymethasone ointment, vaseline and hydrobase were applied. Transepidermal water loss (TEWL) was checked after 0, 3, 6, 9, 12, 15, 18, 21 and 24 hours. Electron microscopic examination was performed after 3 and 24 hours after desoxymethasone, vaseline and hydrobase had been applied. RESULTS: The recovery of TEWL was most prominantly observed in the desoxymethasone ointment treated group followed by vaseline and hydrobase. Electron microscopic examination using ruthenium tetroxide fixation revealed that secretion and numbers of lamellar bodies and complete formatice of lipid bilayers were most prominent at desoxymethasone ointment and vaseline treated group. CONCLUSION: Desoxymethasone ointment, vaseline and hydrobase can be good agents in improving bamer recovery after exposure to irritant material.
Animals ; Benzalkonium Compounds* ; Desoximetasone* ; Irritants ; Lipid Bilayers ; Mice ; Mice, Hairless* ; Permeability ; Petrolatum* ; Ruthenium ; Skin*

BACKGROUND: The epidermal permeability barrier necessary for terrestrial life resides in the intercellular spaces of the stratum corneum and is composed of lipids. OBJECTIVE: Since strrtum corneum lipid may be important for the permeability barrier, we studied the differences and effects of experimentally altered barrier function using acetone and tape-stripping technique. METHODS: The permeability barrier of hairless mouse was disrupted by tape-stripping and acetone treatment and the recovery rate was assessed by histochemical staining, electron microscopic examination and lipid analysis. RESULTS: Although the transepidermal water loss recovered completely by 48 hours in both of the acute models, acetone treated samples seem to have on over-all better recovery rate than tape-stripped samples. The return of barrier function to normal in both tape-stripped and acetone-treated skin was accompanied by a comparable return of normal nile red and ruthenium tetroxide staining. The amount of lipid in stratum corneum paralleled both the return of barrier function towards normal and the extent of prior damage to the barrier in acetone treated skin, yet, the lipid synthesis in tape-stripped skin showed a slower return of lipid content. CONCLUSION: The difference in the recovery rate of the two acute models may be due to the fact that acetone mainly extracts intercellular lipids, whereas, tape-stripping has a prolonged effect by removal of comeocyte in addition to the intercellular lipids. This shows the importance of comeocytes as well as the intercellular lipid bilayer in the recovery of normal barrier function.
Acetone* ; Animals ; Extracellular Space ; Lipid Bilayers ; Mice ; Mice, Hairless ; Permeability ; Ruthenium ; Skin ; Water

BACKGROUND: Moisturizers induce skin hydration and then increase flexibility and elasticity, making the skin soft and smooth, and protecting it against environmental stimuli. OBJECTIVE: The aim of this work was to study the role of intercorneocyte lipid layers and desmosomes in the mechanism of moisturization. METHODS: Transepidermal water loss (TEWL) and capacitance were measured and the morphologic changes of the intercorneocyte lipid layers and desmosomes with electron microscopy, using ruthenium tetroxide (RuO4) postfixation, following the application of glycerin, propylene glycol, and a mixture of glycerin and propylene glycol for a 2 hour period to the epidermis of hairless mice were measured. RESULTS: 1. The TEWL was significantly increased in all three groups; glycerin, propylene glycol, and mixture of glycerin and propylene glycol. The increase of TEWL after the application of glycerin was maintained from the second to the forth hour after application which was statistically significant, after the application of propylene glycol it was maintained for 5 hours, and after the application of a mixture of glycerin and propylene glycol, for 6 hours. 2. The capacitance also was increased in all three experimental groups, compared to the control group. However there was no statistical significance. 3. Light microscopic findings showed no specific changes in all three groups, compared to the control group. 4. Ultrastructural observation by electron microscope, using RuO4 postfixation, showed widening of the intercorneocyte lipid layers in all three groups. In contrast to glycerin in which the results showed detachment of the desmosomes without changes in the intercorneocyte lipid layers, propylene glycol showed interruption and undulation of the intercorneocyte lipid layers and expansion of the lacunae spaces. A mixture of glycerin and propylene glycol showed interruption and undulation of the intercorneocyte lipid layers, detachment of the desmosomes, and, partial, formation of lacunae. CONCLUSION: These results suggest that the moisturizing effects of glycerin result from an increased detachment of the desmosomes and widening of the intercorneocyte lipid layers and then an increase in the water holding capacity of the stratum corneum. Propylene glycol, a chemical penetration enhancer, induce widening, interruption, and undulation of the lipid layers and expansion of the lacunae space. In the mixture of glycerin and propylene glycol, propylene glycol potentiate and continue the moisturizing effects of the glycerin.
Animals ; Desmosomes* ; Elasticity ; Epidermis ; Glycerol* ; Mice ; Mice, Hairless ; Microscopy, Electron ; Pliability ; Propylene Glycol* ; Ruthenium ; Skin