Stapled anoplin peptide combined with photothermal therapy enhances oncolytic immunotherapy of triple-negative breast cancer / 中国中药杂志

This study constructed a nano-drug delivery system, A3@GMH, by co-delivering the stapled anoplin peptide(Ano-3, A3) with the light-harvesting material graphene oxide(GO), and evaluated its oncolytic immunotherapy effect on triple-negative breast cancer(TNBC). A3@GMH was prepared using an emulsion template method and its physicochemical properties were characterized. The in vivo and in vitro photothermal conversion abilities of A3@GMH were investigated using an infrared thermal imager. The oncoly-tic activity of A3@GMH against TNBC 4T1 cells was evaluated through cell counting kit-8(CCK-8), lactate dehydrogenase(LDH) release, live/dead cell staining, and super-resolution microscopy. The targeting properties of A3@GMH on 4T1 cells were assessed using a high-content imaging system and flow cytometry. In vitro and in vivo studies were conducted to investigate the antitumor mechanism of A3@GMH in combination with photothermal therapy(PTT) through inducing immunogenic cell death(ICD) in 4T1 cells. The results showed that the prepared A3@GMH exhibited distinct mesoporous and coated structures with an average particle size of(308.9±7.5) nm and a surface potential of(-6.79±0.58) mV. The encapsulation efficiency and drug loading of A3 were 23.9%±0.6% and 20.5%±0.5%, respectively. A3@GMH demonstrated excellent photothermal conversion ability and biological safety. A3@GMH actively mediated oncolytic features such as 4T1 cell lysis and LDH release, as well as ICD effects, and showed enhanced in vitro antitumor activity when combined with PTT. In vivo, A3@GMH efficiently induced ICD effects with two rounds of PTT, activated the host's antitumor immune response, and effectively suppressed tumor growth in 4T1 tumor-bearing mice, achieving an 88.9% tumor inhibition rate with no apparent toxic side effects. This study suggests that the combination of stapled anoplin peptide and PTT significantly enhances the oncolytic immunotherapy for TNBC and provides a basis for the innovative application of anti-tumor peptides derived from TCM in TNBC treatment.

GPER inhibitor increases tamoxifen-induced apoptosis in T-47DTR-resistant cells / 中国药理学通报

Aim To study the effect of G protein-coupled estrogen receptor(GPER)inhibitor G15 on the sensitivity of breast cancer tamoxifen-resistant cells to T-47DTR. Methods Experiments were carried out with 4-hydroxytamoxifen(4-OHT),the active form of tamoxifen in vivo. The sensitivity of tamoxifen-resistant breast cancer cell line T-47DTR and its parental cell line T-47D to tamoxifen was detected by MTT assay; the expression of GPER protein was analyzed by plasma separation of inhibitor G15; the effect of 4-OHT combined with G15 on the apoptosis of T-47DTR cells was analyzed by flow cytometry AnnexinV-FITC/PI double staining; the expression levels of apoptosis-related proteins Bax,Bcl-2,caspase-3,cleaved caspase-3,caspase-9,cleaved caspase-9 were analysed by Western blot. Results(1)Compared with the parental cell T-47D,the resistance of T-47DTR-resistant cells to 4-OHT was significantly enhanced.(2)When 4-OHT(2 μmol·L-1)was administered,the membrane distribution of GPER increased,indicating that GPER was activated in T-47DTR-resistant cells compared with the control group; Compared with OHT,the use of G15(5 μmol·L-1)and OHT significantly reduced the expression of GPER.(3)GPER inhibitor G15 could increase the apoptotic rate of T-47DTR-resistant cells while down-regulating the anti-apoptotic protein Bcl-2 and up-regulating the expression of pro-apoptotic proteins Bax,cleaved caspase-3,cleaved caspase-9. Conclusions The GPER inhibitor G15 increases the apoptosis of T-47DTR cells and restores the sensitivity of drug-resistant cells to tamoxifen.

Simultaneous quantitation of cytokinin bases and their glycoconjugates with stable isotope labelling ultrahigh performance liquid chromatography mass spectrometry.

Cytokinins (CKs) are one class of important phytohormones widely investigated in most aspects of plant life. Similar to other phytohormones, CKs and their glycoconjugates are hydrophilic. Their ionization efficiencies for mass spectrometry (MS) detection are rather poor, whereas their retention and separation on reverse phase liquid chromatography (RPLC) are often unsatisfying. Chemical isotope labelling LC-MS analysis methods have been developed for most other phytohormones, enhancing their LC separations and quantitative sensitivity. However, there are currently no reports for chemical-labelled CKs. Here, we report a new chemical isotope labelling LC-MS analytical method for one-pot derivatization of CK bases and their glycoconjugates, based on differential benzylation labelling of the adenine skeleton of CKs with benzyl bromide and its deuterium isotope-labelled reagent. Benzylation alters the hydrophilicity of CKs and their glycoconjugates, improving their retention and separation on RPLC. The developed method demonstrated enhanced sensitivity, as the CKs and their glycoconjugates could be analysed with LODs within the range of 0.62-25.9 pg/mL. The method also demonstrated good intra- and inter-day precisions with standard deviations in the range of 1.9%-13.0%, and acceptable accuracy with recoveries in the range of 84.0%-119.9%. The developed method was employed on the quantitation of CKs in the fresh roots of Astragalus membranaceus collected from both fertilized and unfertilized fields. The significant impact that fertilizers had on endogenous CKs metabolism was observed. As such, monitoring endogenous CKs and their metabolites might be promising to control fertilizer abuse.

Research on anti-tumor natural product diosgenin / 中国中药杂志

Diosgenin is widely distributed in many plants, such as Polygonatum sibiricum, Paris polyphylla, Dioscorea oppositifolia, Trigonella foenum-graecum, Costus speciosus, Tacca chantrieri, which has good anti-tumor activity and preferable effects on preventing atherosclerosis, protecting the heart, treating diabetes, etc. This review combed through the anti-tumor mechanisms of diosgenin encompassing lung, breast, gallbladder, liver, oral cavity, stomach, bladder, bone marrow, etc. Besides, it was discovered that diosgenin mainly exerts its effect by inhibiting tumor cell migration, suppressing tumor cell proliferation and growth, and inducing cell apoptosis. However, problems like low yield and bioavailability frequently exist in natural diosgenin. This review introduced methods such as structural modification, dosage form optimization and combination medication to improve the yield and anti-tumor activity of diosgenin. Via the summary of this paper, it is expected to provide theoretical basis for the rational exploitation and utilization of diosgenin.

Biological activities and chemical components of Potentilla kleiniana Wight & Arn.

In this study, we investigated the antimicrobial, antioxidant, and cytoprotective activities of ethanol extract and the ethyl acetate (EtOAc) fraction of P. kleiniana Wight & Arn. The EtOAc fraction exhibited antimicrobial effects against most of the microorganisms that were tested, including Staphylococcus aureus, Candida albicans, Pseudomonas aeruginosa, and, Escherichia coli, but not Aspergillus niger. In addition to its excellent antioxidant activity, the EtOAc fraction attenuated the UVB-induced cell death via upregulation of caspase-3 expression in human keratinocytes. The HPLC/ESI-MS/MS analysis allowed identification of the components in the EtOAc fraction. Overall, our results suggest that P. kleiniana is a valuable source of bioactive compounds for the development of pharmaceuticals and cosmetics.

Correlations of serum sympathetic activity markers with sleep and cognition in patients with chronic insomnia / 中华神经科杂志

Objective:To explore the changes of serum levels of copeptin and α-amylase and their correlations with sleep and cognition in patients with chronic insomnia (CI).Methods:From September 1, 2018 to May 31, 2019, fifty CI outpatients or inpatients from the Department of Sleep Disorder, Affiliated Chaohu Hospital of Anhui Medical University, were enrolled continuously, and thirty good sleepers from the Physical Examination Center of the hospital, were also enrolled to serve as controls. Pittsburgh Sleep Quality Index (PSQI), polysomnography (PSG) and Pre-Sleep Arousal Scale (PSAS) were used to assess the insomnia severity and sleep disorder susceptibility. Montreal Cognitive Assessment scale (MoCA) and Nine-Box Maze were used to respectively assess general cognition and memories. The serum levels of copeptin and α-amylase were detected using enzyme linked immunosorbent assay.Results:Compared to the controls, the CI patients had increased PSQI score (16.0 (15.0, 17.0) vs 4.0 (2.8, 6.0); Z=-7.678, P<0.001) and PSAS score (33.0 (30.0, 37.5) vs 17.0 (16.0, 18.5); Z=-7.350, P<0.001), decreased MoCA score (24.1±2.5 vs 26.7±1.9, t=-4.625, P<0.001), increased numbers of errors in the object working (1.0 (0, 1.0) vs 0 (0, 1.0), Z=-2.099, P=0.036), spatial working (2.0 (1.0, 4.0) vs 1.0 (0, 2.0), Z=-3.935, P<0.001) and object recognition (1.0 (0, 2.0) vs 0 (0, 0), Z=-2.266, P=0.023) memories, and elevated serum levels of copeptin ((35.1±19.9) pg/ml vs (14.8±6.9) pg/ml, t=5.414, P<0.001) and α-amylase ((990.1±193.7) U/L vs (728.9±230.5) U/L, t=5.597, P<0.001). In the CI patients, the level of copeptin was positively correlated with PSQI score ( r=0.338, P=0.013), PSAS score ( r=0.316, P=0.021), sleep latency ( r=0.324, P=0.018), number of awake ( r=0.325, P=0.017) and stage 1 percent of non-rapid eye movement sleep ( r=0.278, P=0.044), and negatively correlated with stage 2 percent of non-rapid eye movement sleep ( r=-0.279, P=0.043); α-amylase was positively correlated with numbers of awake in PSG ( r=0.293, P=0.033). Multiple linear regression analysis showed that copeptin level affected PSQI score (β=0.255, P=0.043) and sleep latency (β=0.254, P=0.043). Conclusion:The levels of copeptin and α-amylase in CI patients elevate, and copeptin may be associated with initial sleep difficulties, but not with cognitive ability, in patients with CI.

Changed serum levels of neurotrophic factors and their correlations with sleep and cognition in patients with chronic insomnia disorder / 中华神经科杂志

Objective To explore serum levels of brain-derived neurotrophic factor (BDNF) and glial-derived neurotrophic factor (GDNF),and whether changes of BDNF and GDNF are correlated with sleep quality and cognitive function in patients with chronic insomnia disorder (CID).Methods Fifty-seven CID patients in the Department of Sleep Disorders,Chaohu Hospital of Anhui Medical University and 30 healthy controls were enrolled from May 2017 to July 2018.Pittsburgh Sleep Quality Index (PSQI) was used to assess the degree of insomnia severity (some CID patients were monitored by overnight polysomnography).Montreal Cognitive Assessment (MoCA) scale and Nine-Box Maze were used to assess general cognitive function and specific memory function,respectively.The serum levels of BDNF and GDNF were detected using ELISA.Results Compared to the controls,CID patients had significantly higher PSQI scores (CID patients:14.0±2.2,healthy controls:3.9± 1.1;t=28.093,P＜0.01),lower MoCA scores (CID patients:24.5±3.6,healthy controls:26.5±0.9;t=-2.985,P＜0.01),more errors in object working memory(CID patients:1.0 (0,1.0),healthy controls:0 (0,0.3)),spatial working memory (CID patients:3.0 (2.0,4.0),healthy controls:1.0 (1.0,2.0)) and object recognition memory (CID patients:0 (0,0),healthy controls:0 (0,0);Z=-2.896、-5.007、-2.306,P＜0.05),and lower serum BDNF (CID patients:(19.48 ± 7.50) ng/ml,healthy controls:(46.49± 13.33) ng/ml;t=-10.274,P＜0.01) and GDNF (CID patients:(32.76± 14.04) pg/ml,healthy controls:(59.63±20.30) pg/ml;t=-7.240,P＜0.01).The partial correlation analysis showed that in the CID patients,the levels of BDNF and GDNF were correlated with PSQI scores negatively (r=-0.293,-0.320,P＜0.05) and MoCA scores positively (r=0.331,0.295,P＜0.05).The BDNF level was also correlated with the duration of disease and the errors in the spatial working memory test negatively (r=-0.319,-0.393,P＜0.05),and the GDNF level was correlated with the total sleep time detected with polysomnogram positively (r=0.520,P＜0.05).Conclusion Serum BDNF and GDNF levels in CID patients were lower than those in healthy controls,and correlated with sleep quality and cognitive impairment.

Changed serum levels of neurotrophic factors and their correlations with sleep and cognition in patients with chronic insomnia disorder / 中华神经科杂志

Objective@#To explore serum levels of brain-derived neurotrophic factor (BDNF) and glial-derived neurotrophic factor (GDNF), and whether changes of BDNF and GDNF are correlated with sleep quality and cognitive function in patients with chronic insomnia disorder (CID).@*Methods@#Fifty-seven CID patients in the Department of Sleep Disorders, Chaohu Hospital of Anhui Medical University and 30 healthy controls were enrolled from May 2017 to July 2018. Pittsburgh Sleep Quality Index (PSQI) was used to assess the degree of insomnia severity (some CID patients were monitored by overnight polysomnography). Montreal Cognitive Assessment (MoCA) scale and Nine-Box Maze were used to assess general cognitive function and specific memory function, respectively. The serum levels of BDNF and GDNF were detected using ELISA.@*Results@#Compared to the controls, CID patients had significantly higher PSQI scores (CID patients: 14.0±2.2, healthy controls: 3.9±1.1; t=28.093, P<0.01), lower MoCA scores (CID patients: 24.5±3.6, healthy controls: 26.5±0.9; t=-2.985, P<0.01), more errors in object working memory (CID patients: 1.0 (0, 1.0), healthy controls: 0 (0, 0.3)), spatial working memory (CID patients: 3.0 (2.0, 4.0), healthy controls: 1.0 (1.0, 2.0)) and object recognition memory (CID patients: 0 (0, 0), healthy controls: 0 (0, 0); Z=-2.896、-5.007、-2.306, P<0.05), and lower serum BDNF (CID patients: (19.48±7.50) ng/ml, healthy controls: (46.49±13.33) ng/ml; t=-10.274, P<0.01) and GDNF (CID patients: (32.76±14.04) pg/ml, healthy controls: (59.63±20.30) pg/ml; t=-7.240, P<0.01). The partial correlation analysis showed that in the CID patients, the levels of BDNF and GDNF were correlated with PSQI scores negatively (r=-0.293, -0.320, P<0.05) and MoCA scores positively (r=0.331, 0.295, P<0.05). The BDNF level was also correlated with the duration of disease and the errors in the spatial working memory test negatively (r=-0.319, -0.393, P<0.05), and the GDNF level was correlated with the total sleep time detected with polysomnogram positively (r=0.520, P<0.05).@*Conclusion@#Serum BDNF and GDNF levels in CID patients were lower than those in healthy controls, and correlated with sleep quality and cognitive impairment.

Changed serum levels of neurotrophic factors and their correlations with sleep and cognition in patients with chronic insomnia disorder / 中华神经科杂志

Objective@#To explore serum levels of brain-derived neurotrophic factor (BDNF) and glial-derived neurotrophic factor (GDNF), and whether changes of BDNF and GDNF are correlated with sleep quality and cognitive function in patients with chronic insomnia disorder (CID).@*Methods@#Fifty-seven CID patients in the Department of Sleep Disorders, Chaohu Hospital of Anhui Medical University and 30 healthy controls were enrolled from May 2017 to July 2018. Pittsburgh Sleep Quality Index (PSQI) was used to assess the degree of insomnia severity (some CID patients were monitored by overnight polysomnography). Montreal Cognitive Assessment (MoCA) scale and Nine-Box Maze were used to assess general cognitive function and specific memory function, respectively. The serum levels of BDNF and GDNF were detected using ELISA.@*Results@#Compared to the controls, CID patients had significantly higher PSQI scores (CID patients: 14.0±2.2, healthy controls: 3.9±1.1; t=28.093, P<0.01), lower MoCA scores (CID patients: 24.5±3.6, healthy controls: 26.5±0.9; t=-2.985, P<0.01), more errors in object working memory (CID patients: 1.0 (0, 1.0), healthy controls: 0 (0, 0.3)), spatial working memory (CID patients: 3.0 (2.0, 4.0), healthy controls: 1.0 (1.0, 2.0)) and object recognition memory (CID patients: 0 (0, 0), healthy controls: 0 (0, 0); Z=-2.896、-5.007、-2.306, P<0.05), and lower serum BDNF (CID patients: (19.48±7.50) ng/ml, healthy controls: (46.49±13.33) ng/ml; t=-10.274, P<0.01) and GDNF (CID patients: (32.76±14.04) pg/ml, healthy controls: (59.63±20.30) pg/ml; t=-7.240, P<0.01). The partial correlation analysis showed that in the CID patients, the levels of BDNF and GDNF were correlated with PSQI scores negatively (r=-0.293, -0.320, P<0.05) and MoCA scores positively (r=0.331, 0.295, P<0.05). The BDNF level was also correlated with the duration of disease and the errors in the spatial working memory test negatively (r=-0.319, -0.393, P<0.05), and the GDNF level was correlated with the total sleep time detected with polysomnogram positively (r=0.520, P<0.05).@*Conclusion@#Serum BDNF and GDNF levels in CID patients were lower than those in healthy controls, and correlated with sleep quality and cognitive impairment.

Atractyligenin, a terpenoid isolated from coffee silverskin, inhibits cutaneous photoaging.

Ultraviolet (UV) light exposure-induced photoaging of the skin is a multifactorial process involving both extrinsic and intrinsic cellular mechanisms. Several naturally occurring products are known to confer protection against UV light-induced skin damage. Our preliminary studies confirmed that the ethyl acetate fraction of coffee silverskin exhibits inhibitory effects on matrix metalloproteases (MMPs). Furthermore, we previously isolated and identified atractyligenin, which has MMP-inhibitory activity, from the silverskin ethyl acetate fraction. The aim of this study was to elucidate the anti-photoaging effects of atractyligenin on human dermal fibroblasts and the underlying mechanism. Human dermal fibroblasts were exposed to 8 J/cm2 UVA radiation, and cell viability was analyzed by MTT assay. The fluorescent dye 2', 7'-dichlorodihydrofluorescein diacetate (H2DCF-DA) was used to measure the intracellular reactive oxygen species (ROS) levels. Our study showed that atractyligenin significantly suppressed the expression of UVA-induced MMPs by inhibiting intracellular ROS production. Atractyligenin treatment reduced c-Jun phosphorylation and c-Fos expression by inhibiting the mitogen-activated protein kinase (MAPK) signaling pathway activated by UVA irradiation. Additionally, treatment with atractyligenin contributed to the homeostasis of collagen by restoring the loss of collagen absorption-related receptor Endo180 and altered fibroblast morphology induced by UVA irradiation. These results indicate that atractyligenin isolated from coffee silverskin inhibits multiple pathways in the human skin photoaging process and is thus a potential candidate for treatment or prevention of photoaging.

Cosmeceutical activities of ethanol extract and its ethyl acetate fraction from coffee silverskin.

BACKGROUND: Coffee silverskin is a thin film that covers the raw coffee bean. In general, coffee silverskin, which detaches during the coffee roasting process, is disposed as firelighters or dispatched to landfills and can cause serious environmental pollution. The aim of this study was to investigate the feasibility of using coffee silverskin as a functional material in cosmetics by evaluating its bioactive ingredients, antioxidative activity, cytoprotective effect, matrix metalloproteinase-1 (MMP-1)-inhibiting effect, and anti-melanogenesis effect. RESULTS: To this end, a 50% ethanol (EtOH) extract and its ethyl acetate (EtOAc) fraction were prepared from coffee silverskin; caffeine was found to be the major compound in the extract. Both the 50% EtOH extract and its EtOAc fraction exhibited antioxidant activities. However, the EtOAc fraction showed a greater radical-scavenging activity and reducing power than that shown by the 50% EtOH extract. Furthermore, the EtOAc fraction increased cell viability in a UVB-irradiated human keratinocyte injury model and significantly suppressed UVB-induced MMP-1 expression and α-melanocyte-stimulating hormone (α-MSH)-stimulated melanin production in HaCaT keratinocytes and B16F1 melanocytes, respectively. Interestingly, caffeine, the major component of the EtOAc fraction, did not show an inhibitory effect. Thus, the antioxidant capacity of the coffee silverskin extract may be attributable to some compounds that exhibit a high antioxidant capacity even at low concentrations or the total antioxidant capacity of various constituent phenolic compounds. CONCLUSION: Our findings indicate that coffee silverskin has the potential for application as a natural functional material in multifunctional cosmetics.

Anti-Aging Activity of Lavandula angustifolia Extract Fermented with Pediococcus pentosaceus DK1 Isolated from Diospyros kaki Fruit in UVB-Irradiated Human Skin Fibroblasts and Analysis of Principal Components.

The effects of Lavandula angustifolia extract fermented with Pediococcus pentosaceus DK1 on UVB-mediated MMP-1 expression and collagen decrease in human skin fibroblasts were determined, and the conversion of its components was also analyzed. Fermentation was performed at varying L. angustifolia extract and MRS medium concentrations, and optimal fermentation conditions were selected. L. angustifolia extracts showed decreased cytotoxicity after fermentation in the fibroblasts. UVB-irradiated fibroblasts treated with fermented L. angustifolia extract showed MMP-1 expression 8.2-14.0% lower than that in UVB-irradiated fibroblasts treated with non-fermented extract. This was observed even at fermented extract concentrations lower than those of non-fermented extracts. Fibroblasts treated with fermented L. angustifolia extract showed 20% less reduction in collagen production upon UVB irradiation than those treated with non-fermented extracts. UVB-irradiated fibroblasts treated with fermented L. angustifolia extracts showed 50% higher inhibition of ROS generation than those treated with non-fermented extract. Luteolin and apigenin glycosides of L. angustifolia were converted during fermentation, and identified using RP-HPLC and LC/ESI-MS. Therefore, the effects of L. angustifolia extract on MMP-1 expression and collagen decrease in UVB-irradiated human skin fibroblasts were increased through fermentation by P. pentosaceus.

Antimelanogenic and Antimigration Properties of the Ethyl Acetate Fraction of Calendula officinalis Flowers on Melanoma Cells.

Calendula officinalis L., commonly known as marigold, is not only cultivated for ornamental purposes but is also used as a traditional medicinal herb. Its flowers have been used to treat various skin diseases, including rashes, burns, cuts and bruises, since ancient times. However, to our knowledge, the impact of C. officinalis L. on melanoma and its mechanism have not been clarified. The aim of this work was to investigate the chemical characterization and antimelanogenic and antimigration activities of the ethyl acetate fraction of C. officinalis flowers (EFC), as well as elucidate the potential mechanism. The obtained results showed that EFC markedly decreased α-MSH-induced melanin production and the cell migration ability of melanoma cells in a dose-dependent manner. Additionally, EFC significantly inhibited the activity and expression of matrix metalloproteinase 2 (MMP-2) via suppressing the mitogen-activated protein kinase (MAPK) signaling pathway. Taken together, the present study demonstrated that C. officinalis flowers can be used as a natural source of antimelanogenisis and antimigration regent to treatment or prevent skin diseases.

Methyl-2-acetylamino-3-(4-hydroxyl-3,5-dimethoxybenzoylthio)propanoate suppresses melanogenesis through ERK signaling pathway mediated MITF proteasomal degradation.

BACKGROUND: Microphthalmia-associated transcription factor (MITF) is regulated by expression and/or degradation pathway, controlling to the expression of melanogenic enzymes for melanin synthesis. Methyl-2-acetylamino-3-(4-hydroxyl-3,5-dimethoxybenzoylthio)propanoate (MAHDP) is reported to anti-melanogenesis effect but its mechanism remain unclear. OBJECTIVE: To investigate the effects of MAHDP on melanogenesis and elucidate its mechanism. METHODS: Tyrosinase activity, melanogenic proteins and gene expression levels were measured with MAHDP treatment in B16F1 cells, human melanocytes, reconstructed skin and clinical trial. RESULTS: MAHDP attenuated melanin production in α-MSH (melanocyte stimulating hormone) stimulated-B16F1 cells. MAHDP decreased the expression of tyrosinase, tyrosinase-related protein-1 (TRP-1), and tyrosinase-related protein-2 (TRP-2). But, MADPH did not affect the phosphorylation of p38 MAPK, JNK and AKT, which are associated with the regulation of MITF expression. These results suggest that MITF downstream is regulated not transcriptionally but translationally. Treatment of MG132 (a proteasomal degradation inhibitor) almost abolished the decrease of MITF protein levels by MAHDP. Phosphorylation and ubiquitination of MITF for proteasomal degradation were increased by treatment of MAHDP. Treatment of PD98059 (an ERK phosphorylation inhibitor) abrogated ERK phosphorylation, downregulation of MITF and tyrosinase as well as the decrease of melanin contents by MAHDP. Therefore, the degradation of MITF proteins by MAHDP is regulated to the ERK signaling. Finally, MAHDP improved the pigmentation in human epidermal melanocytes, a UVB-irradiated the reconstructed skin model and clinical trial without cytotoxicity and skin irritation. CONCLUSION: These results clearly demonstrate that MAHDP suppresses the expression of melanogenic enzymes through ERK phosphorylation-mediated MITF proteasomal degradation, and suggest that MAHDP may be efficient as a therapeutic agent for hyperpigmentation.

Suppression of Ultraviolet B-mediated Matrix Metalloproteinase Generation by Sorbus commixta Twig Extract in Human Dermal Fibroblasts.

Sorbus commixta is a traditional oriental medicinal plant that grows in East Asian countries such as Korea, Japan and China. The twig of S. commixta has been considered valuable for centuries to treat diseases including asthma, cough and other bronchial disorders. However, the effect of S. commixta twig extract on human skin has not been investigated well. The present study aimed at assessing the antiphotoaging effect of S. commixta twig ethanol extract (STE) on ultraviolet B (UVB)-induced matrix metalloproteinase (MMP) levels and its underlying mechanism in human dermal fibroblasts. In this study, we found that STE (12.5-50 µg mL-1 ) treatment significantly inhibited UVB-induced MMP-1, MMP-2 and MMP-3 expression, concomitant with a downregulation of intracellular ROS generation. These effects might be associated with a STE-induced inhibition of the mitogen-activated protein kinase (MAPK) pathway. Furthermore, STE also downregulated UVB-induced c-Fos expression in a concentration-dependent manner, but had no inhibitory effect on c-Jun phosphorylation. Taken together, these results indicate that STE may be an antiphotoaging agent and that its effect may occur via its inhibition of MMPs expression and MAPK pathway activation.

Effect of 5-hydroxytryptamine-2C subunit receptor on long-term potentiation in primary visual cor-tex of form deprivation adult amblyopia rats / 新乡医学院学报

Objective To investigate the effects of 5-hydroxytryptamine-2C subunit receptor (5-HT 2C R)on long-term potentiation (LTP)of V1M region visual cortex of form deprivation adult amblyopia rats. Methods Sixteen two weeks old Sp-argue Dawley rats were randomly divided into normal control group and monocular form deprivation group,with 8 rats in each group. The rats in the normal control group were not given any intervention;the rats in the monocular form deprivation group were sutured the right eye lid to establish the monocular form deprivation amblyopia model. All rats were fed for 6 weeks after establishing the model successfully,then the rats in the two groups were sacrificed and the coronal examination of 400 μm thick cortical brain slices were incubated in artificial cerebrospinal fluid artificial cerebrospinal fluid (ACSF). According to the difference of drugs in ACSF,the visual cortex slices of rats in normal control group were selected as group A;the contralateral visual cortex slices of the deprivation eye were divided into group B,group C,group D and group E;the ipsilateral visual cortex slices of the deprivation eye were divided into group F,group G,group H and group I. The ACSF of group A,B and F did not added any drugs;the ACSF of group C and group G were added with physiological saline;the ACSF of group D and group H were added with 10 μmol · L - 15-hydroxytryptamine hydrochloride;the ACSF of group E and group I were added with 10 μmol·L - 1 SB 242084 and 10 μmol·L - 15-hydroxytryptamine hydrochloride. The electrophysiology experiment was per-formed in all of the visual cortex slices by extracellular microelectrode recording and the visual cortex fidd postsynaptic poten-tial(fPSP)slope of V1M region of the visual cortex was recorded. Results The fPSP in group A,B,C,D,E,F,G,H,I was (198. 1 ± 13. 5)％,(106. 3 ± 8. 3)％,(106. 3 ± 8. 3)％,(157. 1 ± 9. 7)％,(102. 6 ± 4. 7)％,(144. 5 ± 2. 9)％,(144. 5 ± 2. 9)％,(192. 2 ± 8. 6)％ and (129. 7 ± 13. 5)％,respectively. There was statistic difference in fPSP slope of visual cortex a-mong the group A,B,F(P < 0. 001);the fPSP slope of visual cortex of rats in group A was significantly higher than that in the group B and group F(P < 0. 001);the fPSP slope of visual cortex of rats in group B was significantly lower than that in the group F(P < 0. 001). The fPSP slope of visual cortex in group D was significantly higher than that in the group C (t = - 10. 833,P < 0. 001);the fPSP slope of visual cortex in group H was significantly higher than that in the group D and group G(t = - 6. 841,- 10. 616;P < 0. 001). The fPSP slope of visual cortex in group E was significantly lower than that in the group D and group I(t = 11. 872,- 3. 910;P < 0. 001,P < 0. 05);the fPSP slope of visual cortex in group I was signifi-cantly lower than that in the group H(t = 9. 911,P < 0. 001). Conclusion Monocular deprivation can lead to the dysfunction of bilateral visual cortex neurons and 5-hydroxytryptamine hydrochloride can reverse this phenomenon through 5-HT2C R.

The effect of dehydroglyasperin C on UVB-mediated MMPs expression in human HaCaT cells.

BACKGROUND: The ultraviolet B (UVB) from solar radiation increases the generation of reactive oxygen species (ROS), which mediate the production of matrix metalloproteinases (MMPs), and acts mainly on the epidermis layer of the skin. This study was aimed at assessing the anti-photoaging effects of dehydroglyasperin C isolated from Glycyrrhiza uralensis Fisch on MMPs levels in HaCaT human keratinocytes and to elucidate the underlying mechanism. METHODS: The cell viability was measured by MTT assay. Expression, phosphorylation and enzymatic activity of the protein were examined using ELISA, Western blot or gelatin zymography. Intracellular ROS measurement was evaluated by fluorescent ELISA and 2',7'-dichlorodihydrofluorescein diacetate (H2DCF-DA) assay. RESULTS: In the present study, we found that dehydroglyasperin C markedly inhibited UVB-mediated expression of collagenase (MMP-1) and gelatinase (MMP-9) by inhibiting ROS generation. Dehydroglyasperin C treatment also decreased the UVB irradiation-mediated activation of mitogen-activated protein kinase (MAPK), c-Jun phosphorylation, and c-Fos expression. In addition, the down-regulation of UVB-induced c-Jun phosphorylation caused by dehydroglyasperin C treatment was more than the down-regulation of c-Fos expression in the HaCaT human keratinocytes. CONCLUSION: Our results indicated that dehydroglyasperin C may function as a potential anti-photoaging agent by inhibiting UVB-mediated MMPs expression via suppression of MAPK and AP-1 signaling.

Antioxidative and Antiaging Activities and Component Analysis of Lespedeza cuneata G. Don Extracts Fermented with Lactobacillus pentosus.

Lespedeza cuneata G. Don is a traditional herb that has been associated with multiple biological activities. In this study, we investigated the antioxidative/antiaging activities and performed an active component analysis of the non-fermented and fermented (using Lactobacillus pentosus) extracts of Lespedeza cuneata G. Don. The antioxidative activities of the fermented extract were higher than those of non-fermented extracts. The elastase inhibitory activity, inhibitory effects on UV-induced MMP-1 expression, and ability to promote type I procollagen synthesis were investigated in Hs68 human fibroblasts cells. These tests also revealed that the fermented extract had increased antiaging activities compared with the non-fermented extract. A component analysis of the ethyl acetate fractions of non-fermented and fermented extracts was performed using TLC, HPLC, and LC/ESI-MS/MS to observe changes in the components before and after fermentation. Six components that were different before and after fermentation were investigated. It was thought that kaempferol and quercetin were converted from kaempferol glucosides and quercetin glucosides, respectively, via bioconversion with the fermentation strain. These results indicate that the fermented extract of L. cuneata G. Don has potential for use as a natural cosmetic material with antioxidative and antiaging effects.

Study on the relationship between the expression level of serum zinc α2 glycoprotein and type 2 diabetes mellitus complicated with hyperuricemia / 中国医师杂志

Objective To investigate the relationship between the expression level of serum zinc α2 glycoprotein (ZAG) and type 2 diabetes mellitus (T2DM) complicated with hyperuricemia (HUA).Methods A total of 220 patients with T2DM and normal uric acid (group A),100 patients with T2DM complicated with hyperuricemia (group B),and 80 cases of normal healthy people (group C) were randomly selected.The clinical and biochemical indexes and expression levels of serum ZAG among three groups were determined and compared.The relationship between expression level of serum ZAG and related indexes was analyzed with Pearson correlation and multiple linear stepwise regression analysis.Results The serum ZAG levels showed significant differences among groups A,B and C [(58.89 ± 23.17)mg/L vs (86.42 ± 25.36) mg/L vs (40.65 ± 16.48) mg/L] (P ＜ 0.05).Pearson correlation analysis showed that serum ZAG level was positively correlated with body mass index (BMI),triglyceride (TG),glycosylated hemoglobin (HbAlc),creatinine (Cr),urea nitrogen (BUN),and serum uric acid (SUA) (P ＜ 0.05),and negatively correlated with high density lipoprotein cholesterol (HDL-C) (P ＜ 0.05).Multiple linear stepwise regression analysis showed that BMI,TG and HDL-C were independent factors influencing serum ZAG level.Conclusions Serum ZAG level is significantly higher in patients with T2FM,especially in patients complicated with HUA.The increased level may be related to obesity and dyslipidemia,showing that serum ZAG may be involved in the pathogenesis of T2DM complicated with HUA.

Digital anatomical analysis of drilling position of the clavicle in coracoclavicular ligament reconstruction / 中国组织工程研究

BACKGROUND:Coracoclavicular ligament reconstruction with transclavicular-transcoracoid driling is an effective surgical technique to treat acromioclavicular dislocation. A good driling in the clavicle leads to a perfect bony tunnel and a good surgery. OBJECTIVE: To observe the effects of different driling positions of the clavicle on the location of bony tunnels in coracoclavicular ligament reconstruction. METHODS:Sixty three-dimensional digital models of the clavicle and coracoid process were constructed by Mimics13.0. Virtual transclavicular-transcoracoid bony tunnels were established according to different surgical planes with different driling positions in the clavicle. Parameters of these bony tunnels were measured, and the safety was evaluated. Option 1: The driling was made 30 mm distal to the clavicle, located in the center of the front and rear edges of the clavicle surface. Option 2: The driling was made 40 mm distal to the clavicle, located in the center of the front and rear edges of the clavicle surface. Option 3: The driling was made at the straight line of tapered nodule tip and the midpoint of the base of the coracoid process, located at the rear edge of the clavicle upper surface. RESULTS AND CONCLUSION: Bony tunnels in option 1 were extremely on the inside of the coracoid. Bony tunnels in options 1 and 2 were not in the center of clavicle. Bony tunnels in option 3 were in the center of both clavicle and coracoid. The method of locating the driling position with a certain distance to the distal clavicle leads to different results in man’s and woman’s models. To ensure that the bony tunnel can pass through the center of clavicle and coracoid, it is suggested to dril at the straight line of tapered nodule tip and the midpoint of the base of the coracoid process and nearby the rear edge of the clavicle upper surface.