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Objective To investigate the role of adenosine kinase (ADK) small interfering RNA (siRNA) modified corneal endothelial cells on the proliferation and secretion of regulatory T cells (Treg cells).Methods The experiment was divided into transfected group and non-transfected group,FAM-ADK siRNA was transfected into corneal endothelial cells with Lipofectamine 3000 in transfected group,and the average mass concentration of adenosine was detected by high performance liquid chromatography in both groups.The experiment was divided into control group (transfected with control siRNA),rapamycin group (added 10 ng/ml rapamycin to culture supernatant after transfected with control siRNA) and ADK siRNA group (added 10 ng/ml rapamycin to culture supernatant after transfected with ADK siRNA).TUNEL staining was used to detect the proportion of apoptosis.Flow cytometry was used to detect the expressions of intercellular adhesion molecule-1 (ICAM-1),vascular cell adhesion molecule-1 (VCAM-1) and E-selectin in corneal endothelial cells.The cells were divided into single cultured group (lymphocytes were cultured separately),co-culture group (lymphocytes were co-cultured with corneal endothelial cells) and ADK siRNA group (lymphocytes were co-cultured with ADK siRNA transfected corneal endothelial cells),and the proportion of CD4+ CD25+ Foxp3 + Treg cells was detected by flow cytometry,and the content of interleukin-10 (IL-10) and transforming growth factor-β (TGF-β) in the supernatant was detected by ELISA in the three groups.The access and use of clinical specimens was approved by Ethic Committee of Tianjing Taida Hospital (TD201705140901).Results Flow cytometry showed that 95.1％ of the endothelial cells expressed siRNA.The average mass concentration of adenosine in the supernatant of the transfection group was significantly higher than that in the control group ([38.020±6.658] ng/ml vs.[1.663 ±0.581] ng/ml) (t =5.437,P =0.006).TUNEL staining showed that the average apoptotic cell proportion of corneal endothelial cells in the rapamycin group was significantly higher than that in the control group,and the average apoptotic cell proportion of corneal endothelial cells in the ADK siRNA group was significantly lower than that in the rapamycin group,with significant differences between them (t =3.763,P =0.020;t =4.405,P =0.012).Flow cytometry showed that the average fluorescence intensities of ICAM-1,VCAM-1 and E-selectin in the ADK siRNA group was weaker than that in the control group (4.060± 1.179 vs.11.600±2.427,3.600 ± 1.234 vs.11.030 ± 2.291,5.223 ± 1.734 vs.24.270 ± 4.332),with significant differences between them (t =2.794,P =0.049;t =2.857,P =0.046;t =4.081,P =0.015).The proportions of Treg cells in the single cultured group,co-culture group and ADK siRNA group was significantly different (F =12.890,P =0.007),and the proportion of Treg cells in the ADK siRNA group was significantly lower than that in the co-culture group (t =3.650,P =0.022).ELISA assay showed that,the contents of IL-10 and TGF-β in the supernatant in the single cultured group,co-culture group and ADK siRNA group were significantly different (F =20.960,P =0.003;F =27.320,P=0.001),and the content of IL-10 and TGF-β in the supernatant in the ADK siRNA group was significantly higher than that in the co-culture group,respectively (t =4.492,P =0.011;t =5.280,P =0.006).Conclusions ADK siRNA modified corneal endothelial cells can induce Treg cells to proliferate and secrete IL-10 and TGF-β,which provides a new method for induction of corneal allograft immune tolerance.
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Objective To discuss the application value of new non-invasive dynamic monopole radiofrequency (RF) system in body shaping.Methods 26 patients were included in this study,all female,aged 31 to 56 years.Application of new type of dynamic monopole radio frequency therapeutic apparatus (UK BTL Exilis therapeutic apparatus) was used to treat abdominal fat,proud flesh between the waist,back,hips,thighs and legs,upper arm area shape and the skin four times,with each time interval of 7～10 days.The patients filled in the subjective preliminary assessment questionnaire of satisfaction.Results All the patients had varying degrees of improvement in relaxation and partial fat reduction,with 76.9％ of satisfactory rate.No adverse reactions and no skin burns occured.Conclusions The new non-invasive dynamic monopole RF system can be used to improve the degree of relaxation and local fat reduction in the body molding,with high satisfaction.
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Objective To discuss the long-term curative effect of botulinum toxin A(BTXA) for treating spasm of eyelid and facial muscle.Methods Ninty-two patients with spasm of eyelid and facial muscle were treated with BTXA and observed for a long term.Results After being injected with BTXA for 1～4 days, the spasm relieved gradually. Sixteen weeks later, the spasm symptoms relapsed. The relapse began mostly from eyelid spasm. The facial spasm's relief time was longer. lt might be up to 24 weeks. After injection, the patients had some slight complications of facial expression numb, ptosis and fissura palpebrae. The above complication symptoms recovered by themselves after two weeks. Conclusion The curative effect of BTXA on treating spasm of eyelid and facial muscle is reliable.
