1.Explicit physical self and its correlation with their implicit physical self in college students
Youyun XU ; Yating QIAN ; Kan HE
Chinese Mental Health Journal 2017;31(2):162-167
Objective:To investigate the relationship between explicit and implicit physical self among contemporary college students.Methods:A total of 485 college students were recruited,with 264 male students,221 female students,266 junior students,219 senior students,189 students in the urban areas and 296 students in the rural areas.The Adolescents'Physical Self Scale (APS) was used to measure their explicit physical self.The Implicit Association Tests were designed to examine implicit physical self of 50 college students,who were selected from 485 college students by isometric random sampling.Results:The difference were significant among the five dimensions of college students' APS scores (P < 0.001).The average scores of college students' appearance,sexual attractiveness were the highest,while the average score of body flaws was the lowest.Male students scored higher in five APS dimensions than female students (Ps < 0.05).Senior students scored higher in the appearance,movement characteristics,figure and body flaws than junior students (Ps <0.05).Students in the urban areas scored higher in the appearance,movement characteristics,figure and body flaws than the students in the rural areas(Ps < 0.05).There was no significant correlation between explicit and implicit physical self (P > 0.05).The scores of relative separation index of movement characteristics were lower in male students than in female students (Z =2.45,P < 0.05).The scores of relative separation index of body flaws were lower in the urban ones than in the rural ones (Z =3.14,P < 0.01).Conclusion:It suggests that separation phenomenon exists between explicit and implicit physical self and the separation phenomena is distinct among the students of different genders or from different areas.
2.Effect of siRNA targeting survivin gene on the apoptosis of malignant melanoma cell line A375
Siyuan CHEN ; Jing CHENG ; Changzheng HUANG ; Yue QIAN ; Yating TU
Chinese Journal of Dermatology 1995;0(04):-
Objective To study the effect of siRNA targeting survivin gene on the apoptosis of malignant melanoma cell line A375. Methods The eucaryotic expression vector of pU-survivin-siRNA was constructed and transfected into the A375 cells by electroporation. The protein expression of survivin was examined by Western blotting, and cell apoptosis by flow cytometry. Results The transfection of pU-sur-vivin-siRNA significantly down-regulated survivin expression ( 0.24 ?0.02 in the transfected group versus 0.98 ?0.21 in the control group ) in A375 cells, and promoted cell apoptosis ( 83% in the transfected group versus 28% in the control group, P
3.Targeted killing of malignant melanoma cells by aclarubicin liposome conjugated with vascular endothelial growth factor
Hongxiang CHEN ; Qiang TONG ; Yue QIAN ; Yan WU ; Aiping FENG ; Zhihong WU ; Xiaofeng YAN ; Yating TU
Chinese Journal of Dermatology 2008;41(7):429-432
Objective To evaluate the targeted killing of malignant melanoma cells by aclarubicin liposomes conjugated with vascular endothelial growth factor(ADM-VEGF-SSL)in vitro.Metheds To detect the binding abilitv of liposomes to malignant melanoma(MM)cells,the human malignant melanoma cell line A375 was cultured in the presence of ADM-VEGF-3H-SSL or ADM-3H-SSL for 2 days followed by the detection of radioactivity of these cells.Then.A375 cells were cultured with various concentrations(0.01,0.1,1,10,100 mol/L)of ADM-VEGF-SSL,ADM-SSL or free ADM for 48 hours in the 48-hour cytotoxity test,or for 0.5 hour followed by another 48-hour culture in drug-free medium in the 0.5-hour cytotoxity test.After that,MTT assay was used to detect the survival rate of these cells.Results ADM-VEGF-SSL could specifically bind to and kill A375 cells.The binding rate of ADM-VEGF-SSL was 2.15 folds as high as that of ADM-SSL.The survival rate of A375 cells after being treated with ADM-VEGF-SSL for 48 hour was similar to that with flee ADM(P>0.05).but lower than that with ADM-SSL(P<0.05),while the survival rate of melanocytes treated with ADM-VEGF-SSL was higher than that with free ADM or ADM-SSL(both P<0.05).As shown by the 0.5-hour cytotoxity test.shortening the treatment course did not attenuate the effect of ADM-VEGF-SSL on A375 cells.Conclusions ADM-VEGF-SSL can specifically recognize A375 cells.efficiently deliver adriamycin into tumor cells,markedly inhibit the proliferation of A375 cells,and eventually,a targeted kill of these cells is realized.
4.The regional genomic instability induced by 60Co γ-rays in B16 cells transfected by GFP
Jing LIU ; Yating WANG ; Hai LIN ; Qian HAN ; Chunxiao ZHANG ; Ou BAI
Chinese Journal of Radiological Medicine and Protection 2012;32(5):465-468
Objective To detect the regional genomic instability of B16 cells treated with 60Co γ-rays by a green fluorescence protein (GFP)-based genomic instability reporting system.Methods Three groups were employed as non-transfection group,vector control group and transfection group.The GFP-marked reporter construct pCMV-EGFP2XhoI for regional genomic instability was successfully transfected into B16 cells using liposome.B16 cells were selected by screening of G418 with a series of concentrations and limiting dilution cultures to yield a single colony.B16 cells with the genomic instability report system were then irradiated by 60Co γ-rays at doses of 0,2 and 4 Gy.The regional genomic instability of B16 cellswas quantified by counting the number of cells with GFP expression.Results B-16 cell strain steadilyexpressing the GFP-based genomic instability reporting system was established successfully.GFP-positiveB16 cells were observed at 1 d after irradiation with 60Co γ-rays at doses of 2 and 4 Gy.Positive correlations between fluorescence intensity and dose and fluorescence intensity and time were also observed.The positive expression rate of GFP followed the increased of dose (F =36.55,36.76,P < 0.05) and time (t =-3.27,-3.16,-4.26,-6.11,-7.17,P < 0.05),and differences between groups were significant.The positive expression rate of GFP increased significantly at 3 d,and maximum expression was observed at 5 d(2.46 ± 0.24 and 3.82 ± 0.35).The level was tending towards stability.Spontaneous GFP expression at a ratio of 1/600 000 was observed in 0 Gy group after 2 weeks of culture.Conclusions The regional genomic instability of B16 cells induced by 60Co γ-rays can be detected using a GFP-labelled genomic instability reporter system.
5.Proliferating T cells and dendritic cells in the skin lesions of patients with mycosis fungoides
Aiping FENG ; Jing ZHAO ; Yan WU ; Yue QIAN ; Siyuan CHEN ; Xin LIAN ; Yating TU
Chinese Journal of Dermatology 1995;0(04):-
Objective To evaluate the profile of proliferating T cells and dendritic cells ( DCs ) in the skin infiltrates of patients with mycosis fungoides ( MF) in different stages. Method Paraffin section and immunohistochemisty with monoclonal antibody were used to detect the expression of special antigen per section. Results The numbers of Ki-67+ cells and cutaneous lymphocyte-associated antigen ( CLA+) cells both increased significantly in the skin infiltrates of MF. Most Ki-67+ cells expressed both CLA and CD4 antigen. The number of Ki-67+ cells was significantly higher ( P
6.Cutaneous intravascular NK/T cell lymphoma: a case report
Siyuan CHEN ; Liduan ZHENG ; Yan WU ; Ying XIA ; Yue QIAN ; Li ZHU ; Cuiyan WANG ; Lan SUN ; Yating TU ; Changzheng HUANG
Chinese Journal of Dermatology 2012;45(3):151-154
An 18-year-old female presented with painful erythema and nodules on both legs for more than 2 years.Dermatological examination showed irregularly sized,mildly indurated,tender,deep subcutaneous nodules arising in diffused infiltrated dark erythematous patches in the inner and posterior region of the left leg.Histopathology showed no significant changes in the epidermis.There were perivascular lymphoid cell infiltrates in the dermis and subcutis.Multiple sites of necrosis of blood vessel walls with vascular occlusions were noted.The lumens of some blood vessels were filled with lymphocytes,among which were many atypical cells with hyperchromatic nuclei and pathologic mitotic figures.Immunohistochemistry showed that lymphocytes in the cavities of blood vessels were positive for CD3(+++),CD3ε(+++),CD2(+),CD56(+++),granzyme B(+++),perforin(+++),CD30(+),Ki67 (+++,100% ),but negative for CD20,CD5,CD7,CD4,CD8,TdT,anaplastic lymphoma kinase,early membrane antigen (EMA) or pan cytokeratin (pCK).The endothelial cells lining the blood vessels stained positively for CD34.The intravascular lymphocytes were also positive for EBER1/2 by in situ hybridization.A diagnosis of cutaneous intravascular NK/T cell lymphoma was made.
7.Characteristics of posterior corneal astigmatism and aberration in cataract patients with high myopia
Qinghe JING ; Yating TANG ; Dongjin QIAN ; Yi LU ; Yongxiang JIANG
Chinese Journal of Experimental Ophthalmology 2018;36(5):360-367
Objective This study was to investigate the characteristics of posterior corneal astigmatism (PCA) and aberration in cataract patients with high myopia.Methods A retrospective study was designed.Two hundred and eighty-two eligible eyes of 190 cataract patients were enrolled in Eye and ENT Hospital of Fudan University from September to December,2014.The eyes were classified into two groups according to axial length (AL):high myopia group with 139 eyes (AL≥26 mm) and control group with 143 eyes (AL was 20 to 25 mm).The mean keratometric mid-radius of curvature (Km),corneal central thickness (CCT),astigmatism and aberrations were measured by the rotating Scheimpflug System (Pentacam),and the AL were measured by the partial coherence interferometry (IOL Master).This study followed the Helsinki declaration,and was approved by the Ethic Committee of Eye and ENT Hospital,Fudan University.Informed consent was signed from each patient.Results In high myopia group,the mean PCA was 0.3 D (range 0 ~ 0.9 D) and 92.8% eyes had PCA values <0.5 D.The steep corneal meridian was aligned vertically (60°~ 120°) in 87.1% eyes for the posterior corneal surface.There was no significant difference in PCA between the high myopia group and the control group (P =0.797).Significant positive linear correlations was found between PCA and anterior corneal astigmatism (ACA),PCA and anterior corneal root mean square (RMS),PCA and anterior lower-order RMS,PCA and posterior corneal RMS,PCA and posterior high-order RMS,PCA and posterior lower-order RMS (r =0.235,P =0.005;r =0.217,P =0.010;r =0.229,P =0.007;r =0.395,P =0.000;r =0.243,P =0.004;r =0.384,P =0.000).Compared with total corneal astigmatism (TCA),anterior corneal measurements overestimated with-the-rule astigmatism (WTR) by a mean of (0.27 ± 0.18) D in 65.67% eyes,underestimated against-the-rule astigmatism (ATR) by (0.27 ± 0.18) D in 88.10% eyes and underestimated oblique astigmatism (Obl) by (0.22 ± 0.10) D in 63.33% eyes.Compared with total corneal aberrations,anterior corneal aberrations measurements overestimated by (0.275 ±0.176) μm in 87.05 % eyes,and the anterior corneal astigmatism types had no effect on the result.Conclusions In high myopia group,92.8% eyes had PCA values <0.5 D and the main astigmatism type in posterior corneal surface was ATR.The posterior corneal astigmatism and aberration were needed to consider in choosing intraocular lens (IOL) before cataract surgery.
8.Establishment of Lymphangioma Model and a Study on the Promoting Effect of Murine Melanoma Cell B16-F1 on the Lymphangiogenesis In Vitro
Siyuau CHEN ; Aijun CHEN ; Chaugzheng HUANG ; Yue QIAN ; Zhixiang LIU ; Yan WU ; Yating TU
Journal of Huazhong University of Science and Technology (Medical Sciences) 2007;27(6):733-735
To establish an animal model of benign lymphangiomas of C57BL/6 mouse in vitro and to observe the effect of mouse ascites melanoma cell B16-F1 on the lymphangiogenesis, 16 C57BL/6 mice aged 8 weeks were given two intraperitoneal injections of incomplete Freund's adjuvant at a15-day interval. The induced neoplasms were studied histopathologically and thhe neoplasms speci- mens were immunohistochemically examined for the expressions of VEGF-C (vascular endothelial growth factor-C) and Fit-4 (VEGFR-3, vascular endothelial growth factor receptor-3). The neoplasms were harvested and embedded in fibrin gel for culture in conditioned medium of B16-F1 cells in vitro and observed under inverted microscope. Our results showed that white solid tumor masses devel- oped in peritoneal cavity after the induction. The tumors were confirmed to be lymphangioma by gross and histological examination. The tumor cells expressed both VEGF-C and Flt-4. Lymphatic capillaries coming from lymphangioma specimen grew into the gel and the conditioned medium of B16-F1 cells was found to be able to promote the growth of the vessels. It is concluded that intrap- eritoneal injection of incomplete Freund's adjuvant is a good method for inducing benign lymphan- giomas in mouse and B16-F1 cells can promote lymphangiogenesis.
9.Construction and identification of LSD1 overexpression and demethylation disfunction plasmids
Lili WU ; Yijie LIU ; Bingyan CAO ; Qiaoman FEI ; Xiujuan ZHAO ; Qian LI ; Yating HAN ; Lei CAO ; Bing YANG ; Ling ZHANG
International Journal of Biomedical Engineering 2018;41(1):26-31,37
Objective To construct rat lysine-specific histone demethylase 1 (LSD1) overexpression plasmids and LSD 1 demethylation fragment disfunction plasmids,and to evaluate their expression levels in HEK293T cells.Methods LSD1 fragments were amplified by PCR,and LSD1 demethylation disfunction fragments were amplified by overlap PCR.Rat-specific LSD1 overexpression plasmids and LSD1 demethylation disfunction plasmids were constructed and verified by agarose gel electrophoresis and sequencing.HEK293T cells were infected using the validated recombinant plasmids and blank vectors,and the stably expressed cells were selected by puromycin.The expression of LSD1 in the stably expressed HEK293T was detected by Western Blot and real-time quantitative PCR.Results The results of agarose gel electrophoresis and sequencing showed that LSD1 overexpression plasmids and LSD1 demethylation disfunction plasmids were successfully constructed.The Western Blot and real-time quantitative PCR results showed that compared with the blank group,the relative expression of LSD1 and mRNA in the LSD1 overexpression group and the LSD1 demethylation disfunction group were up-regulated,and the differences were statistically significant(all P<0.01).Conclusions The constructed LSD1 overexpression plasmids and LSD1 demethylationi disfunction plasmids can achieve overexpression of LSD1 gene in HEK293T cells.This paper lays a foundation for further study of the relationship between LSD 1 gene and related diseases and demethylation of LSD 1.
10.CRISPR-Cas9-based site-directed knock-in of VEGF165 gene in a HEK293T cell
Zaiyu GUO ; Heliang ZHANG ; Qian CHEN ; Yanwei HOU ; Tao SHUI ; Lili WU ; Yijie LIU ; Qiaoman FEI ; Huan HUANG ; Lei LEI ; Yan SUN ; Yu KONG ; Xiujuan ZHAO ; Yating HAN ; Bing YANG ; Ling ZHANG
International Journal of Biomedical Engineering 2019;42(1):39-44
Objective To construct a human renal epithelial cell line HEK293T by CRISPR-Cas9-based site-directed knock-in of vascular endothelial growth factor 165 (VEGF165) gene, and avoid the off-target effect caused by lentivirus infection. Methods The VEGF165 expression vector with homologous arm (pUCm-T-VEGF165 plasmid) and the sgRNA expression vector [pSpCas9(BB)-2A-Puro-sgRNA plasmid] were designed and constructed based on the DNA sequence of the EZH2 gene, and then co-transfected into HEK293T cells. The expression of VEGF165 mRNA was detected by qPCR and the expressions of VEGF165 proteins were detected by Western Blot. Results The qPCR and Western Blot results showed that, comparing with the control, the pUCm-T-VEGF165 plasmid and pSpCas9(BB)-2A-Puro-sgRNA plasmid, the expression of the co-transfection plasmid were significantly increased, i.e. 3.42±0.30 vs. 1.02±0.21, 1.13±0.16 and 0.98±0.18 for the VEGF165 mRNA level (all P<0.01), and 1.13±0.16 vs. 1.02±0.06, 0.88±0.03 and 0.80±0.05 for the VEGF165 protein level (all P<0.01), respectively. Besides, the expression of EZH2 was significantly down-regulated, i.e. 0.14±0.06 vs. 1.08±0.11, 1.02±0.12 and 1.13±0.16 for the EZH2 mRNA level (all P<0.01), and 0.23±0.03 vs. 1.05±0.13, 0.91±0.04 and 0.81±0.06 for the EZH2 protein level (all P<0.01), respectively. This result showed that the VEGF165 was successfully inserted into the EZH2 genome, interfering the EZH2 expression. Conclusions VEGF165 gene can be successfully knocked into HEK293T cells by CRISPR/Cas9 system.