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Objective:To preliminarily investigate the effect of circIGF2BP3 on autophagy in photoaged dermal fibroblasts.Methods:Human dermal fibroblasts were isolated from circumcised foreskin tissues from 6 children in the Department of Urological Surgery, Third Affiliated Hospital of Sun Yat-sen University. An ultraviolet A (UVA) -induced photoaged human dermal fibroblast model (UVA radiation group) was established by repeated UVA radiation at a dose of 10 J/cm 2 for 14 consecutive days, and human dermal fibroblasts receiving no treatment served as control group. The photoaged cell model was verified by β-galactosidase staining, Western blot analysis for determining P21 protein expression, and cell counting kit-8 (CCK8) assay for evaluating cell viability. Moreover, Western blot analysis was performed to determine the protein expression of autophagy-related proteins P62, microtubule-associated protein 1 light chain 3 (LC3) -Ⅰand LC3-Ⅱ in photoaged human dermal fibroblasts, and real-time quantitative RCR (qRT-PCR) to verify the differential expression of circIGF2BP3 between photoaged and normal human dermal fibroblasts. Furthermore, circIGF2BP3 was biologically annotated. Some cultured primary human dermal fibroblasts were divided into 4 groups: empty vector group transfected with an empty vector, UVA + empty vector group transfected with an empty vector followed by repeated UVA radiation, circIGF2BP3 group transfected with a circIGF2BP3-overexpressing lentiviral vector, UVA + circIGF2BP3 group transfected with a circIGF2BP3-overexpressing lentiviral vector followed by repeated UVA radiation. Western blot analysis was performed to determine the expression of autophagy-related proteins. Statistical analysis was carried out by using t test, one-way analysis of variance and least significant difference- t test. Results:Compared with the control group, the UVA radiation group showed significantly increased proportions of β-galactosidase-positive cells (61.33% ± 5.78% vs. 6.37% ± 0.32%, t = 9.49, P < 0.01) and P21 expression (1.25 ± 0.03 vs. 1.00 ± 0.05, t = 4.26, P < 0.05), but significantly decreased cell viability (74.33% ± 3.48% vs. 100%, t = 7.38, P < 0.01). Moreover, the P62 expression and LC3-Ⅱ/Ⅰ ratio were significantly higher in the UVA radiation group than in the control group (both P < 0.05). The relative expression of circIGF2BP3 was 0.72 ± 0.04 in the photoaged human dermal fibroblasts, which was significantly lower than that in the normal human dermal fibroblasts (1.00 ± 0.03, t = 5.46, P < 0.01). The P62 expression and LC3-Ⅱ/Ⅰ ratio were significantly lower in the circIGF2BP3 group (0.60 ± 0.01, 0.71 ± 0.01, respectively) than in the empty vector group (1.00 ± 0.02, 1.00 ± 0.01, t = 16.25, 2.75, P < 0.01, < 0.05, respectively), and lower in the UVA + circIGF2BP3 group (1.05 ± 0.02, 2.04 ± 0.05, respectively) than in the UVA + empty vector group (1.31 ± 0.02, 2.72 ± 0.14, t = 10.493, 6.472, respectively, both P < 0.01) . Conclusion:circIGF2BP3 can regulate autophagy in UVA-induced photoaged dermal fibroblasts, which provides a new potential therapeutic target for the prevention and treatment of skin photoaging.
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Objective To explore the developmental characteristics of white matter volume in autism spectrum disorder (ASD) children longitudinal.Methods From May 2011 to September 2014,37 ASD children (ASD group)and 27 developmental delays (DD) children (control group) were treated at the Child Mental Health Research Center,Nanjing Brian Hospital Affiliated of Nanjing Medical University,and the children whose age,gender and developmental quotient matched with the ASD children were scanned by structure magnetic resonance imaging (sMRI) at the age of 2-3 years old and 4-5 years old respectively.Region of interest (ROI) technology was adopted to investigate the change of the cerebrum white and the sub-lobes structure white matter volume with time.Then the correlation between clinical symptoms and brain white matter volume changes was analyzed.Results Among the 2-3 years old,compared with the control group,the white matter volume of the total brain[(383 521.84 ±6 427.57) mm3 vs.(364 014.06 ±6 856.97) mm3],the left cerebral hemisphere [(191 609.35 ± 3 206.60) mm3 vs.(181 695.89 ± 3 389.54)mm3],temporal lobe [(41 860.49 ±816.38) mm3 vs.(39 444.18 ± 834.85) mm3] and the right temporal lobe [(21 312.79 ± 414.07) mm3 vs.(20 084.22 ± 412.13) mm3] were significantly larger in the ASD group,and the differences were statistically significant (all P ＜ 0.05).With the analysis of covariance with age or the total brain volume as the covariate,the differences disappeared(all P ＞ 0.05).Among the 4-5 years old,compared with the control group,the white matter volumes of the total brain[(417 651.42 ± 6 443.86) mm3 vs.(394 317.27 ± 6 404.86)mm3],left cerebral hemisphere [(208 714.16 ±3 214.61) mm3 vs.(197 192.82 ±3 262.02) mm3],right cerebral hemisphere [(208937.26±3242.09) mm3 vs.(7 124.45 ±3 193.13) mm3],frontal lobe [(107 107.46±1 681.99) mm3 vs.(100 326.19 ± 1 883.24) mm3],left frontal lobe [(54 569.63 ± 846.85) mm3 vs.(51 177.25 ±979.09) mm3],right frontal lobe [(52 537.83 ± 841.99) mm3 vs.(49 148.94 ±928.31) mm3],temporal lobe [(45 189.75 ± 833.29) mm3 vs.(42 487.73 ± 786.27) mm3],left temporal lobe [(22 204.21 ±411.77) mm3 vs.(20 922.90 ± 418.46) mm3],and right temporal lobe [(22 985.54 ± 426.93) mm3 vs.(21 564.83 ± 378.78) mm3]were significantly larger in the ASD group,and the differences were statistically significant (all P ＜ 0.05).With the analysis of covariance with age as the covariate,the differences still existed (all P ＜ 0.05).With the analysis of covariance with the total brain volume as the covariate,the differences disappeared (all P ＞ 0.05).For longitudinal analysis,there was a significant difference in the white matter volume between the whole brain,left cerebral hemisphere,right cerebral hemisphere,frontal lobe,left frontal lobe,fight frontal lobe,temporal lobe,left temporal lobe,right temporal lobe and the differences were statistically significant (F =5.521,5.533,5.459,5.830,5.800,5.723,4.857,4.418,5.159,all P ＜0.05).There was a positive correlation between the changes of the volume of whole brain,the white matter volume in the whole brain,bilateral cerebral hemisphere,frontal lobe,parietal lobe,right parietal lobe and Childhood Autism Rating Scale (r =0.367,0.343,0.321,0.349,0.296,0.308,0.351,all P ＜ 0.05).Conclusions Among the 2-3 years old,the white matter volume of the brain regions have been increased significantly in ASD.Among the 4-5 years old,the increase of the white matter volume of the brain regions implicated more widely which mainly concentrated in the frontal and temporal lobe in ASD.The severity of the clinical symptoms of ASD may be associated with the white matter volume of the total brain,bilateral cerebral hemisphere,frontal lobe,parietal lobe and right parietal lobe.
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Objective To investigate the significant of peripheral CD4+CD69+T lymphocytes in patients with autoimmune hemolytic anemia (AIHA)/Evans syndrome (ES).Methods In this study peripheral blood samples from 32 patients with AIHA/ES (15 hemolytic episode patients,17 remission patients) and 13 healthy controls were collected.Patients with AIHA/ES were recruited in Tianjin Medical University General Hospital from October 2015 to May 2016.The percentages of CD69+ T lymphocytes were analyzed by flow cytometry.The expression of CD69 mRNA in CD4+T lymphocytes which was sorted from peripheral blood by magnetic activated cell sorting (MACS) was detected using real-time PCR.Soluable CD69 was measured by ELISA.Results In hemolytic episode patients,the ratio of CD3+CD69+/CD3+T lymphocytes [(3.08 ± 1.48)％] was significantly higher than that in healthy controls [(1.28 ± 0.83)％,P＜0.01] and in remission group[(1.96± 1.33)％,P＜0.05].The absolute count of CD3+CD69+T lymphocytes in hemolytic episode group [(2.94± 1.81)× 107/L] was higher than that in healthy controls [(1.48± 1.42)× 107/L,P＜0.05].The ratio of CD3+CD4+CD69+/CD3+CD4+T cells in hemolytic episode group [(2.16± 1.56)％] was significantly higher than that in remission group [(1.16±0.62)％,P＜0.05] and healthy controls[(0.94±0.78)％,P＜0.05].The quantity of CD3+CD4+CD69+T lymphocytes in hemolytic episode group[(1.04±0.98)× 107/L] was higher than in healthy controls [(0.44± 0.38) × 107 / L,P＜0.05].The ratio of CD3+CD8+CD69+/CD3+ CD8+T lymphocyte in hemolytic episode group [(4.87±2.56)％] was significantly higher than that in healthy controls[(1.83± 1.27)％,P＜0.01].The quantity of CD3+CDs+CD69+T lymphocytes in three groups did not show significant difference.The ratio of CD3+CD4+CD69+/CD3+CD4+T lymphocytes in hemolytic episode group was negatively correlated with hemoglobin (Hb) (P＜0.01),positively correlated with the percentage of reticulocytes (Ret％)(P=0.01)total bilirubin(TBil),indirect bilirubin(IBil) (P＜0.01) and not correlated with absolute reticulocytes count,lactic dehydrogenase (LDH),complement 3(C3),complement 4 (C4).The ratio of CD3+CD4+CD69+/CD3+CD4+T lymphocytes in remission group was negatively correlated with Hb (P＜0.05).In hemolytic episode patients CD69 mRNA (32.26±35.11) was significantly higher than that in remission group(6.05±5.87)(P＜0.05)and healthy controls (1.76± 1.85)(P＜0.01).CD69 mRNA in remission group was significantly higher than healthy controls (P＜0.05).Serum CD69 in hemolytic episode patients [(494.21 ± 16.06) ng/L] was significantly higher than that in healthy controls [(441.39± 104.6) ng/L,P＜0.05].Conclusion Our findings suggest that the proportion of CD4+CD69+ T lymphocytes increase in AIHA/ES patients,which is correlated with the severity of disease.