BACKGROUNDElastin derived peptides can regulate melanocyte precursor development. Ultraviolet irradiation, infrared radiation and heat can increase the synthesis of tropoelastin in human skin epidermis. The aim of this study was to investigate whether the over expressed tropoelastin in epidermis has some role in melanogenesis of melanocytes.

METHODSA375 human melanoma cells were treated with different concentrations of kappa elastin for 24 hours. A375 human melanoma cells were randomly assigned to control, kappa elastin, and lactose pre-incubated groups. The cell viabilities were detected by the methyl thiazoleterazolium assay. Melanin content and tyrosinase activity in A375 melanoma cells were measured. The expressions of endothelin B receptor (ET(B)R) mRNA and c-kit mRNA in A375 melanoma cells were measured by quantative reverse transcription polymerase chain reaction.

RESULTSFifty µg/ml of kappa elastin significantly increased the melanin content by 56.64% compared with the control (P < 0.05). Kappa elastin increased cellular tyrosinase activity by 46.73% compared with the control at 24 hours (P < 0.05). Kappa elastin increased the expressions of ET(B)R and c-kit mRNA levels by 2.13-fold and 2.47-fold compared with the controls, respectively. When pre-incubating cells with a lactose solution (10 mmol/L), the inhibition on melanin production was 34.96% compared with the kappa elastin group (P < 0.05), tyrosinase activity was inhibited by 29.93% compared with kappa elastin group (P < 0.05), and the expressions of ET(B)R mRNA and c-kit mRNA were decreased by 1.56-fold and 0.82-fold compared with kappa elastin group, respectively.

CONCLUSIONKappa elastin increased the melanogenesis in A375 melanoma cells via the stimulation of tyrosinase activity and the expression of ET(B)R and c-kit. The over expressed tropoelastin produced by keratinocytes might play a role in melanogenesis of epidermal melanocytes.

Cell Line, Tumor ; Cell Survival ; drug effects ; Elastin ; pharmacology ; Humans ; Keratinocytes ; drug effects ; Melanins ; metabolism ; Melanoma ; Proto-Oncogene Proteins c-kit ; metabolism ; Receptor, Endothelin B ; metabolism

BACKGROUNDOxidative stress plays an important role in the pathogenesis of epidermal diseases. This study aimed to investigate the effects of quercetin on the anti-oxidative response and on mitochondrial protection in cultured normal human keratinocytes.

METHODSCultured HaCaT cells were treated with different concentrations of H2O2 (0, 50, 100, 250, 500 micromol/L) for different periods of time (0.5, 1, 2, 4 hours) to establish an oxidative stress model. The cultured HaCaT cells were randomly assigned to control, H2O2, and quercetin + H2O2 groups. For the quercetin groups, the cells were treated with different concentrations of quercetin (0, 10, 25, 50 micromol/L) before exposure to H2O2. Morphological changes of the cells were observed under an inverted microscope and an electron microscope. The cell viability was detected by the MTT method. The cell apoptosis (AnnexinV/propidium iodide double stain) and mitochondrial membrane potential (DeltaPsim) changes were detected by flow cytometry.

RESULTSAn oxidative stress model of HaCaT cells was established under a suitable concentration (250 micromol/L) and treated time of H2O2 (2 hours). The cell viability and DeltaPsim decreased in a concentration-dependent and time-dependent manner while the percentage of apoptotic cells significantly increased in the H2O2 groups compared with the control group (P < 0.05). The cell viability and DeltaPsim of the quercetin treated group increased (P < 0.05) and the percentage of apoptotic cells decreased at concentrations of 1-50 micromol/L quercetin (P < 0.01) compared with H2O2 treated group.

CONCLUSIONQuercetin can relieve the cell damage and apoptosis from H2O2 induced injury to HaCaT cells by anti-oxidation and mitochondrial protection.

Antioxidants ; pharmacology ; Apoptosis ; drug effects ; Cell Survival ; drug effects ; Cells, Cultured ; Dose-Response Relationship, Drug ; Humans ; Hydrogen Peroxide ; toxicity ; Keratinocytes ; drug effects ; pathology ; Membrane Potential, Mitochondrial ; drug effects ; Mitochondria ; drug effects ; Quercetin ; pharmacology

Diacylglycerol, DAG, because of its multifunctional and nutritional properties, attracted considerable attention recently. Enzymatic synthesis of diacylglycerols from linoleic acid was investigated in a solvent-free reaction in a continuously operated fixed bed reactors containing Lipozyme RM IM. By appropriate manipulation of the fluid-residence time, the relative proportions of the various acylglycerols in the effluent stream can be controlled. In addition, the presence of excess glycerol is effective for the removal of water produced during the esterification reactions. Under the conditions of molar ratio of linoleic acid to glycerol of 0.5, the immoblized enzyme maintained high stability and allowed the reaction to continue for 10 days without significant deterioration in enzyme activity. It was determined that the conversion of fatty acid, content of 1,3-DAG and volume efficiency of reactor reached optima under the conditions: a packaged-bed reactor(with a ratio of packed length to inner diameter of 7.8), reacting temperature at 65 degrees C, molar ratio of linoleic acid to glycerol of 0.5, and feeding flow rate of 1.2 mL/min.
Catalysis ; Diglycerides ; chemical synthesis ; Enzyme Stability ; Enzymes, Immobilized ; chemistry ; Lipase ; chemistry

Objective To explore the impact and regularity of snail population after changing of water level and to develop effective control and prediction programs. Methods A typical snail habitat closed to Changjiang county in Yueyang city, Hunan province was selected as the survey field.Data on water level and the changing trend of snail population during 2001-2009 including the average water level, maximum and minimum water levels, snail and infected snail densities,proportion of sampling frames with living and infected snails etc. were collected. The distribution of water level and snail indexes were described and a general additive model(GAM)for the relationships between these indexes were also fitted. Results Impacts of the average water level and the watered-out days in current year in the surveyed field on the snail density were statistically significant(P＜0.01), showing a quadratic curve association. The snail density remained at the lowest level when the average water level was at 25.0 m and the surveyed field watered-out days was 120. The average water level in current year and the field watered-out days in the past year showed statistically significant impacts on the infected snail density(P＜0.001), the proportion of sampling frames with infected snails(P＜0.05)and living snails(P value neared 0.05), presented a quadratic curve, a cubic curve or even a more complicated piecewise curve association. Both the infected snail density and the proportion of sampling frames with infected snails remained at the lowest level when the average water level was at 25.0 m and the watered-out days in the past year was 110, while the proportion of sampling frames with living snails kept the lowest level when the average water level was at 25.0 m and the watered-out days in the past year was 160. Conclusion The water level and the field watered-out days affected the development of snail population directly. The changing water level had an impact on snail population change, which mainly presented as nonlinear smooth function relation. Impact of the field watered out days on the infected snail density and the proportion of sampling frames with living snails and infected snails showed a hysteresis effect. The snail density was predicted to be retaining a high level when the water level was 24.0 m and the field watered-out days was 3 months. It had obvious advantages to fit the relationship of the changing water level and the snail indexes with a GAM which could get closer to the reality as well as easier to find and explain the potential associations and regulations.

To set up a new pattern of quality control and evaluation for Chinese medicine. By investigating the limitation of quality control pattern for Chinese medicine, the differences and similarities in the chemical substantial style as well as quality control patterns among Chinese medicine, chemical synthetic drugs and Biologicals, combining with the author's experience on the research of geo-authentic medicinal material and theory of Chinese medicinal nature, a new pattern of quality control for Chinese medicine has been explored and designed. A more rational pattern of quality control for Chinese medicine should be referred to Biologicals instead of chemical synthetic drugs, there are more similarity in chemical substantial style and quality control pattern for Chinese medicine between Chinese medicine and Biologicals than that between Chinese medicine and chemical synthetic drugs. Based on geo-authentic medicinal material and bioassay or biopotency detection, a new pattern of quality control for Chinese medicine could be built and applied.
Biological Assay ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; standards ; Energy Transfer ; Evaluation Studies as Topic ; Medicine, Chinese Traditional ; standards ; Pattern Recognition, Automated ; Plants, Medicinal ; chemistry ; Quality Control ; Technology, Pharmaceutical ; methods

OBJECTIVETo explore the method for obtaining olfactory ensheathing cells from human fetal olfactory mucosa by cell culture for selective adhesion in the presence of neurotrophin-3 (NT3) and low-concentration serum.

METHODSThe olfactory ensheathing cells were cultured alternatively in DMEM/F12 culture medium containing 10% fetal bovine serum (FBS) and the medium containing NT3 and 2.5% FBS every 72 h. The cells were observed for morphological changes and identified using immunocytochemistry with P75NTR and GFAP, and the cell purity was estimated.

RESULTSThe olfactory ensheathing cells from human fetal olfactory mucosa were positive for P75(NTR) and GFAP, and in in vitro culture, the cells exhibited dipolar or tripolar appearance with long thin neurites. On the 9th day of cell culture, the purity of the olfactory ensheathing cells reached about 83%.

CONCLUSIONThe olfactory ensheathing cells can be obtained by in vitro culture for selective adhesion in the presence of NT3 and low-concentration serum.

Cell Culture Techniques ; methods ; Cell Separation ; methods ; Cells, Cultured ; Culture Media ; Fetus ; Humans ; Neurotrophin 3 ; pharmacology ; Olfactory Bulb ; cytology ; Olfactory Mucosa ; cytology

OBJECTIVETo study the effects of HKL-4 on physiological changes during growth of leaves.

METHODUsing licorice (Glycyrrhiza uralensis) as material, the effects of HKL-4 on active oxygen metabolism and photochemical efficiency in licorice leaf were determined under field condition.

RESULT AND CONCLUSIONThe contents of chlorophyll, activity of SOD and CAT increased, while the MDA contents in leaves decreased. The senescence was delayed, so that the photochemical efficiency (Fv/Fm) was increasing comparing to the control.

Catalase ; metabolism ; Chlorophyll ; metabolism ; Glycyrrhiza ; growth & development ; metabolism ; Malondialdehyde ; metabolism ; Peroxidase ; metabolism ; Photosynthesis ; Plant Growth Regulators ; pharmacology ; Plant Leaves ; growth & development ; metabolism ; Plants, Medicinal ; growth & development ; metabolism ; Superoxide Dismutase ; metabolism

ObjectiveTo investigate the clinical value of color Doppler ultrasound examination in the diagonosis of acute and chronic artery occlusion of the extremities.MethodsA review was made on 129 extremetiy artery occlusion patients at Anzhen Hospital during 2006 -2010. 85 cases were male, and 44 cases were female. Age was from 17 to 94 years (average: 62 ±9 years). We analyzed two-dimensional and color Doppler flow imagings of 39 acute occlusion arteries and 97 chronic occlusion arteries. We compared factors including the echoes of artery lumens, the vessel wall structures, hemodynamic parameters of inlet and outlet at the occlusion, and collaterals between groups.ResultsThe factors of depths of vessel wall,internal diameters of ccclusion arteries, proximal resistant index and collaterals were significantly different between groups ( P ＜ 0. 05 ). The internal diameters of acute occlusion arteries were wider than chronic occlusion arteries. The depths of vessel wall, proximal resistant index and collaterals were thinner, smaller,and less than chronic occlusion arteries. The total accurate rate of differential diagnosis for acute and chronic artery occlusion by color Doppler ultrasound was 95.6％.ConclusionsColor Doppler ultrasound is an effective method for the differential diagnosis of acute and chronic artery occlusion of the extremities.

OBJECTIVETo explore the possibilities of bone marrow stromal cells (MSCs) to adopt Schwann cell phenotype in vitro and in vivo in SD rats.

METHODSMSCs were obtained from tibia and femur bone marrow and cultured in culture flasks. Beta-mercaptoethanol followed by retinoic acid, forskolin, basic-FGF, PDGF and heregulin were added to induce differentiation of MSCs'. Schwann cell markers, p75, S-100 and GFAP were used to discriminate induced properties of MSCs' by immunofluorescent staining. PKH-67-labelled MSCs were transplanted into the mechanically injured rat sciatic nerve, and laser confocal microscopy was performed to localize the PKH67 labelled MSCs in the injured sciatic nerve two weeks after the operation. Fluorescence PKH67 attenuation rule was evaluated by flow cytometry in vitro.

RESULTSMSCs changed morphologically into cells resembling primary cultured Schwann cells after their induction in vitro. In vivo, a large number of MSCs were cumulated within the layer of epineurium around the injured nerve and expressed Schwann cell markers, p75, S-100, and GFAP.

CONCLUSIONMSCs are able to support nerve fiber regeneration and re-myelination by taking on Schwann cell function, and can be potentially used as possible substitutable cells for artificial nerve conduits to promote nerve regeneration.

Animals ; Biomarkers ; analysis ; Bone Marrow Cells ; cytology ; Cell Differentiation ; Cells, Cultured ; Flow Cytometry ; Fluorescent Antibody Technique, Indirect ; Fluorescent Dyes ; Glial Fibrillary Acidic Protein ; analysis ; Morphogenesis ; Organic Chemicals ; analysis ; Phenotype ; Rats ; Receptor, Nerve Growth Factor ; analysis ; S100 Proteins ; analysis ; Schwann Cells ; cytology ; metabolism ; Sciatic Nerve ; cytology ; injuries ; Stromal Cells ; cytology ; metabolism ; transplantation

The study is designed to synthesize nano-carrier Tyr-RGD (cyclo-[Arg-Gly-Asp-d-Tyr-Lys]) and poly(ethylene glycol) modified polyethylenimine (Tyr-RGD-PEG-PEI) targeting vascular endothelial cells, then analyze its nanoparticle properties and the characteristics of drug carrying and targeting properties in vivo / in vitro tumor. The nano-carrier Tyr-RGD-PEG-PEI was synthesized with the method of chemical synthesis and the properties of this nanoparticle and drug carrying characteristics were identified. Its effect of targeting vascular endothelial cells in vitro was studied with the method of competitive binding assay. The fluorescent labeled nano-drug was injected into tumor-bearing nude mice to observe its tumor-targeting. The mean size of nano-carrier Tyr-RGD-PEG-PE was about 145 nm, good in encapsulation efficiency of siRNA. After incubation in plasma for half an hour, only about 3 percent of siRNA out. It was confirmed that it was a single spot with TLC analysis, the R(f) value was 0.65. Receptor competition experiments showed that the nano could effectively compete with RGD in binding the receptors on endothelial cells. Tumor-bearing nude mice experiments showed that when containing a fluorescent-labeled siRNA of Tyr-RGD-PEG-PEI nano-drug was injected into mice, after 24 hours this nano-drug mainly distributed within the tumor tissue. However, nano-drug without Tyr-RGD appeared in tumor tissue as well as other organs such as livers, lungs, etc. The Tyr-RGD-targeted gene vector Tyr-RGD-PEG-PEI synthesized in this study has good nanoparticle properties and high efficiency of gene-drug encapsulation. Study of nude mice shows that the ability of its tumor-targeting is significantly better than nano-drug without Tyr-RGD.
Animals ; Endothelial Cells ; metabolism ; Gene Transfer Techniques ; Genetic Vectors ; Humans ; Integrins ; biosynthesis ; Mice ; Mice, Nude ; Nanoparticles ; Oligopeptides ; chemical synthesis ; pharmacology ; RNA, Small Interfering