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Objective:To compare the efficacy and safety of picosecond alexandrite laser and ultrapulsed fractional CO 2 laser for the treatment of facial atrophic acne scars in a prospective, split-face clinical trial. Methods:From October 2015 to October 2017, patients with facial symmetrical atrophic acne scars were enrolled from Department of Cosmetic Laser Surgery, Hospital for Skin Diseases, Chinese Academy of Medical Sciences and Peking Union Medical College. The left side of the face of the patients was treated with ultrapulsed fractional CO 2 laser, and the right side of the face was treated with a 755-nm picosecond alexandrite laser with a diffractive lens array. All patients received 3 sessions of laser treatment at a 2-month interval. The severity of scars was evaluated by 2 dermatologists before and after the treatment according to the ECCA grading scale (échelle d′évaluation clinique des cicatrices d′acné) , and clinical efficacy was self-assessed by the patients according to a 4-point scale. Pain scores and adverse reactions were recorded. Paired t test was used to compare ECCA scores and pain scores between the 2 sides of the face, as well as between pre- and post-treatment values, and Wilcoxon rank sum test was used to compare the 4-point scores between the 2 groups. Results:There was no significant difference in the ECCA score between the picosecond laser side and the fractional laser side before the treatment ( t = 1.06, P = 0.300) , while the ECCA score was significantly higher in the picosecond laser side than in the fractional laser side after 3 sessions of treatment (70.98 ± 21.48 vs. 58.04 ± 17.63, t = 3.76, P = 0.001) . Compared with the pre-treatment severity score of scars, the improvement in severity score was significantly less in the picosecond laser side than in the fractional laser side (2.21 ± 1.09 vs. 2.83 ± 1.11, z = 2.70, P = 0.007) . Compared with the fractional laser side, the picosecond laser side showed significantly lower pain scores (3.71 ± 0.62 vs. 6.23 ± 1.06, t = 11.93, P < 0.001) and less adverse reactions, which mainly manifested as transient erythema and edema. Conclusions:Both picosecond alexandrite laser and fractional CO 2 laser can effectively improve atrophic acne scars. Fractional CO 2 laser is more effective, but picosecond alexandrite laser has less adverse effects.
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Objective To evaluate the efficacy and safety of a 755 nm picosecond Alexandrite la-ser with a diffractive lens array in the treatment of facial photoaged skin .Methods Twenty-six pa-tients with facial photoaging were recruited and received 3 treatments at 4-week intervals .Laser energy was applied over the entire face at a fixed spot size of 6 mm ,with a fluence of 0 .71 J/cm2 and 5Hz . Blinded clinical assessment was performed by 2 independent dermatologists on a 5-point global pho-toaging scale (GPS) .Patients were also questioned on the extent of improvement of rhytides ,skin tightening ,and complexion with a 4-point global aesthetic improvement scale (GAIS) and satisfaction . Adverse events were also evaluated .Results Twenty-six patients completed the treatment .Compared with the baseline ,there was a significant improvement in facial photoaged skin after 3 treatments ,and these positive outcomes were maintained up to the 3-month follow-up ,according to the GPS and GAIS scores .Moderate pain and transient erythema were observed as the two main discomforts associated with the treatment .Most patients were satisfied with the treatment .Conclusions This 755 nm pico-second Alexandrite laser with a diffractive lens array optic is effective in the treatment offacial pho -toaged skin ,and the therapy also seems safe and well tolerated .
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Objective To evaluate the regulatory effects of lycopene on the key signaling receptors in human cutaneous squamous cell carcinoma cell line COLO16.Methods Cultured COLO16 cells were divided into 6 groups to be treated with lycopene at different concentrations of 0,5,10,15,20,and 25 μmol/L,respectively,for 24 hours (control group and 5,10,15,20,25 μmol/L lycopene groups),followed by estimation of the cell viability by lactate dehydrogenase (LDH) assay.Lycopene at a safe concentration was selected based on the LDH assay,and used for the determination of expression of signaling receptors,and Western blot analysis was performed to measure the expression of key signaling receptor proteins,including epidermal growth factor receptor (EGFR),glucocorticoid receptor (GR),retinoic acid receptor-alpha (RAR-α),retinoid X receptor-alpha (RXR-α),androgen receptor (AR) and progesterone receptor (PR).Statistical analysis was carried out by one-way analysis of variance (ANOVA),Tukey multiple comparison teat and Brown-Forsythe test with the SPSS software.Results After 24-hour treatment with lycopene at different concentrations,there were significant differences in the rate of cell death among these groups (F =13.116,P ＜ 0.05),and the rate of cell death in the 25 μmol/L lycopene group significantly differed from that in the control group (P ＜ 0.05).Therefor,lycopene at concentrations of 5,10 and 20 μmol/L were selected to treat COLO16 and HaCaT cells as well as human epidermal keratinocyte (HEK) for 24 hours in the following experiment.The treatment with lycopene significantly decreased the phosphorylation level of EGFR (P ＜ 0.05),but significantly increased the expression of GR protein (P ＜ 0.05),and showed no significant effects on the protein expression of RAR-α,RXR-α,AR,and PR in COLO16 cells.After 24-hour treatment with lycopene at concentrations of 5,10 and 20 μmol/L,there were no significant changes in the phosphorylation level of EGFR protein or the expression of GR protein in HaCaT cells and HEK (all P ＞ 0.05) compared with those without lycopene treatment.Conclusion Lycopene can decrease the viability of COLO16 cells,inhibit the activation of EGFR protein,and up-regulate the expression of GR,and these effects may be specific for tumor cells.
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Objective To optimize experimental conditions for transfecting primary human melanocytes with Wnt5A-specific siRNA,and to establish the Wnt5A gene silencing model.Methods Primary human melanocytes (PHM)were cultured in vitro.Positive control GAPDH-siRNAs at different concentrations of 0,20.0,33.3,40.0 and 60.0 nmol/L were transfected into PHM by using liposomes.Realtime fluorescence-based quantitative PCR (qPCR) was performed to select the optimal concentration of siRNA with the highest transfection efficiency.Three Wnt5A-siRNAs including Wnt5A-siRNA-793,Wnt5A-siRNA-943 and Wnt5A-siRNA-1743 were constructed and transfected into PHM separately,and qPCR was conducted to select the most specific Wnt5A-siRNA.Then,the most specific Wnt5A-siRNA was transfected into PHM,and the total mRNA and total protein were extracted after 24-,48-and 72-hour treatment.qPCR and Western blot analysis were performed to determine the optimal time with the highest transfection efficiency.Results There were significant differences in the mRNA expression of GAPDH among cells transfected with GAPDH-siRNAs at different concentrations of 0,20.0,33.3,40.0 and 60.0 nmol/L (1.009 ± 0.161,0.086 ± 0.010,0.140 ± 0.016,0.285 ± 0.095,0.012 ± 0.007 respectively;F =69.469,P ＜ 0.05).Additionally,the 60-nmol/L GAPDH-siRNA group showed the lowest GAPDH mRNA expression (all P ＜ 0.05),as well as the best transfection efficiency.The mRNA expression of Wnt5A differed in the Wnt5A-siRNA-793 group,Wnt5A-siRNA-943 group,Wnt5A-siRNA-1743 group and the blank control group (0.331 ± 0.010,2.229 ± 0.029,0.078 ± 0.006 and 1.000 ± 0.024 respectively;F =7 006.094,P ＜ 0.05),and the Wnt5A-siRNA-1743 group showed the lowest Wnt5A mRNA expression (all P ＜ 0.05),as well as the best transfection efficiency.The mRNA expression of Wnt5A differed among the Wnt5A-siRNA-1743 group after 24-,48-and 72-hour treatment and the blank control group (0.396 ± 0.002,0.026 ± 0.008,0.131 ± 0.079,1.025 ± 0.276 respectively;F =29.215,P ＜ 0.05),so did the protein expression of Wnt5A (112.798 ± 0.218,77.765 ± 0.415,30.540 ± 0.219,130.025 ± 0.158 respectively;F =79 122.889,P ＜ 0.05).Moreover,the Wnt5A mRNA expression was significantly lower in the Wnt5A-siRNA-1743 group at 48 hours compared with that at 24 and 72 hours and the blank control group (all P ＜ 0.05),while the Wnt5A protein expression was significantly lower in the Wnt5A-siRNA-1743 group at 72 hours compared with that at 24 and 48 hours and the blank control group (all P ＜ 0.05).Conclusion The siRNA-mediated Wnt5A gene silencing model of human melanocytes is established successfully.
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Objective To optimize experimental conditions for transfecting primary human melanocytes with Wnt5A-specific siRNA,and to establish the Wnt5A gene silencing model.Methods Primary human melanocytes (PHM)were cultured in vitro.Positive control GAPDH-siRNAs at different concentrations of 0,20.0,33.3,40.0 and 60.0 nmol/L were transfected into PHM by using liposomes.Realtime fluorescence-based quantitative PCR (qPCR) was performed to select the optimal concentration of siRNA with the highest transfection efficiency.Three Wnt5A-siRNAs including Wnt5A-siRNA-793,Wnt5A-siRNA-943 and Wnt5A-siRNA-1743 were constructed and transfected into PHM separately,and qPCR was conducted to select the most specific Wnt5A-siRNA.Then,the most specific Wnt5A-siRNA was transfected into PHM,and the total mRNA and total protein were extracted after 24-,48-and 72-hour treatment.qPCR and Western blot analysis were performed to determine the optimal time with the highest transfection efficiency.Results There were significant differences in the mRNA expression of GAPDH among cells transfected with GAPDH-siRNAs at different concentrations of 0,20.0,33.3,40.0 and 60.0 nmol/L (1.009 ± 0.161,0.086 ± 0.010,0.140 ± 0.016,0.285 ± 0.095,0.012 ± 0.007 respectively;F =69.469,P ＜ 0.05).Additionally,the 60-nmol/L GAPDH-siRNA group showed the lowest GAPDH mRNA expression (all P ＜ 0.05),as well as the best transfection efficiency.The mRNA expression of Wnt5A differed in the Wnt5A-siRNA-793 group,Wnt5A-siRNA-943 group,Wnt5A-siRNA-1743 group and the blank control group (0.331 ± 0.010,2.229 ± 0.029,0.078 ± 0.006 and 1.000 ± 0.024 respectively;F =7 006.094,P ＜ 0.05),and the Wnt5A-siRNA-1743 group showed the lowest Wnt5A mRNA expression (all P ＜ 0.05),as well as the best transfection efficiency.The mRNA expression of Wnt5A differed among the Wnt5A-siRNA-1743 group after 24-,48-and 72-hour treatment and the blank control group (0.396 ± 0.002,0.026 ± 0.008,0.131 ± 0.079,1.025 ± 0.276 respectively;F =29.215,P ＜ 0.05),so did the protein expression of Wnt5A (112.798 ± 0.218,77.765 ± 0.415,30.540 ± 0.219,130.025 ± 0.158 respectively;F =79 122.889,P ＜ 0.05).Moreover,the Wnt5A mRNA expression was significantly lower in the Wnt5A-siRNA-1743 group at 48 hours compared with that at 24 and 72 hours and the blank control group (all P ＜ 0.05),while the Wnt5A protein expression was significantly lower in the Wnt5A-siRNA-1743 group at 72 hours compared with that at 24 and 48 hours and the blank control group (all P ＜ 0.05).Conclusion The siRNA-mediated Wnt5A gene silencing model of human melanocytes is established successfully.
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Objective To estimate the effect of a tretinoin derivative ECPIRM on retinoic acid receptors (RARs),and to observe skin irritation responses to it in mice.Methods Cultured SCL-1 cells were divided into 2 groups to be treated with culture medium containing 10 μmol/L ECPIRM (ECPIRM group) or 10 μmol/L all-trans retinoic acid (ATRA) (ATRA group) for 24 hours,and those treated with drug-free culture medium served as the control group.Western blot analysis and real-time fluorescence-based quantitative PCR were performed to quantify the protein and mRNA expressions of RARs (RARα,RARβ,RARγ and RXRα) respectively.In addition,real-time fluorescence-based quantitative PCR was conducted to measure the mRNA expressions of two target genes of the activated RAR signaling pathway,i.e.,cytochrome P450 26A1 (CYP26A1) and tazarotene-induced gene 1 (TIG1).Eight BALB/c mice were equally divided into 2 groups to be topically treated with 0.075％ ECPIRM gel or 0.05％ ATRA cream at equal molar concentrations on the shaved skin once daily for 21 successive days.Skin irritation reactions were assessed in these mice.Results Compared with the control group,the ATRA group showed significantly increased protein and mRNA expressions of RARα,RARβ and RARγ (all P ＜ 0.05).The mRNA expressions of CYP26A1 and TIG1 genes in the ATRA group were 25.49 and 3.88 times that in the control group respectively (both P ＜ 0.01).However,there was no significant difference in the protein expressions of RARα,RARβ,RARγ and RXRα,or mRNA expressions of RARα,RARβ,RARγ CYP26A1 and TIG1 between the ECPIRM group and control group (all P ＞ 0.05).Obvious Skin irritation reactions such as erythema and desquamation were observed in BALB/c mice after 2-day topical treatment with ATRA cream,and their degree peaked after 5-day treatment.However,neither erythema nor desquamation was observed in BALB/c mice during 21-day treatment with 0.075％ ECPIRM gel.Conclusion Unlike ATRA,ECPIRM cannot activate the canonical RAR signaling pathway or cause skin irritation reactions.
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@#Taiji Quan exercise has been confirmed benefit in the rehabilitation for spinal cord injured patients, that may improve the motor and sense function, promote the physical and mental health.
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Objective To investigate the effect of adenovirus-mediated IL-24 (Ad-IL-24)gene expression on the apoptosis in a human squamous cell carcinoma cell line COLO 16, and to explore the underlying molecular mechanism. Methods Cultured COLO 16 cells were divided into two groups to be transfected with an adenovirus vector carrying the IL-24 gene (Ad-IL-24 group)or green fluorescent protein (Ad-GFP group), while those receiving no treatment served as the control group. After culture for different durations, qPCR was performed to quantify IL-24 gene expression, methyl thiazolyl tetrazolium (MTT) assay to evaluate the proliferative activity of COLO 16 cells, flow cytometry to detect the apoptosis of COLO 16 cells, laser scanning confocal microscopy (LSCM) to observe the morphological changes of COLO 16 cells, Western blot to determine the levels of Bax and Bcl-2 proteins and to evaluate the activation of caspase-3, qPCR to determine the levels of Bax and Bcl-2 mRNAs, an immunofluorescence assay to observe the expression of Bax and Bcl-2 proteins. Statistical analysis was carried out by a two-sample t-test with the SPSS 19.0 software. Results MTT assay showed that the proliferation of COLO 16 cells in the Ad-IL-24 group was significantly inhibited as early as 4 days after the transfection; thereafter, the inhibitory effect increased in a time-dependent manner, and peaked on day 6(P0.05). Flow cytometry revealed that the apoptosis rate was significantly higher in the Ad-IL-24 group(13.10%± 0.92%)than in the control group(3.69%± 0.36%, P0.05). LSCM demonstrated that the apoptosis of COLO 16 cells was accelerated in the Ad-IL-24 group. The immunofluorescence assay, Western blot and qPCR all showed that the mRNA and protein expressions of Bax were increased, but those of Bcl-2 were decreased in the Ad-IL-24 group compared with the Ad-GFP group and control group. Moreover, Western blot showed a protein band that could specifically bind to the anti-cleaved caspase-3 antibody in the Ad-IL-24 group, but not in the Ad-GFP group or control group. Conclusions Ad-IL-24 can induce apoptosis in human COLO 16 squamous cell carcinoma cells, probably by up-regulating Bax expression, down-regulating Bcl-2 expression, and activating caspase 3.
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Objective To study the effect of paeonol on the proliferation and apoptosis of A375 human melanoma cells and its mechanism.Methods Cell counting kit-8 (CCK8) was used to evaluate the proliferative activity of A375 cells treated with paeonol of 0.5,1,2,4,8 mmol/L for 24,48 and 72 hours respectively.Subsequently,A375 cells were treated with paeonol of 1.25,2.5 and 5 mmol/L for 24 hours followed by double staining with annexin V and propidium iodide for the detection of cell apoptosis,fluorometric assay for the estimation of caspase 3,caspase 8 and caspase 9 activity,and Western blot for the determination of the levels of p53,nuclear factor-κB proteins and some of their target proteins.The A375 cells receiving no treatment served as the blank control group.Statistical analysis was carried out by t test.Results Within the investigated concentration and time ranges,paeonol significantly inhibited the proliferative activity of A375 cells in a concentration-and timedependent manner.Compared with the blank control group,a significant increase was observed in the early apoptosis rate in A375 cells treated with paeonol of 1.25,2.5 and 5 mmol/L for 24 hours (13.74％-± 1.73％,25.95％ ± 0.57％ and 46.44％ ± 0.81％ vs.3.11％ ± 0.53％,P ＜ 0.05 or 0.01),as well as in the activity of caspase 3,8 and 9 in A375 cells treated with paeonol of 2.5 and 5 mmol/L for 24 hours (P ＜ 0.05 or 0.01).After 24-hour treatment,the protein levels of p53 and Bax were elevated,but those of nuclear factor-κB,Bcl-2 and Bcl-XL were decreased in A375 cells with the increase of paeonol concentration.Conclusions Paeonol can inhibit the proliferation but induce the apoptosis of A375 cells,and the apoptosis-inducing effect may be realized through intrinsic and extrinsic pathways by modulating nuclear factor-κB and p53 genes.