Objective To determine the level of interleukin (IL)-10 in sera and culture supernatants of CD4+CD25+T cells from children with atopic dermatitis (AD),and to evaluate its relationship with clinical course and severity of AD.Methods Totally,46 children with AD and 31 healthy controls were included in the study.All the patients were divided into 3 groups,i.e.,mild (n =10),moderate (n =16) and severe (n =20) group,according to severity scoring of atopic dermatitis (SCORAD) score.Venous blood samples were obtained from the patients and healthy controls.CD4+CD25+ regulatory T cells were separated from the blood samples by magnetic cell sorting (MACS) system in two steps and cultured in vitro.Enzyme linked immunosorbent assay (ELISA) was conducted to quantify the level of IL-10 in sera and culture supernatants of CD4+CD25 + T cells from these subjects.Analysis of variance was carried out to compare the level of supematant and serum IL-10 between the patients and controls,and Pearson correlation analysis to assess the relationship between the level of IL-10 and SCORAD score.Results The patients with mild,moderate and severe AD showed a similar serum IL-10 level compared with the healthy controls ((43.10 ± 25.07) pg/ml,(68.40 ± 36.65) pg/ml and (55.55 ± 41.97) pg/ml vs.(58.27 ± 36.84) pg/ml,all P ＞ 0.05).The level of supernatant IL-10 secreted by CD4+CD25+ T cells from the controls was significantly higher than that from the patients with severe AD ((55.15 ± 11.15) pg/ml vs.(27.25 ± 7.01) pg/ml,P ＜ 0.05),but similar to that from the patients with mild and moderate AD ((52.96 ± 11.69) pg/ml and (49.86 ± 9.18) pg/ml,respectively,both P ＞ 0.05).The level of secreted IL-10 was negatively correlated with SCORAD score (r =－0.757,P ＜ 0.01),whereas the serum level of IL-10 showed no statistical correlation with SCORAD score.Conclusion CD4+CD25+ T cells and IL-10 may be implicated in the development of AD.

Finding stable expression sites on the chromosomes of Chinese hamster ovary (CHO) cells is an effective method to solve the problem of unstable expression of CHO cells in long-term culture. Our group used lentiviral transfection to integrate the tracer gene (Zsgreen1) into the chromosome of CHO cells and found multiple potential stable expression sites. This study verified the ability of one of the sites located in the 148052-148157 bp region on chromosome NW_003614241.1 to stably express exogenous proteins.The expression of Zsgreen1 gene was first observed, and CRISPR/Cas9 technology was then used to integrate the enhanced green fluorescent protein (EGFP) gene into this site. Three strains of EGFP gene integrated cells were obtained. After 60 generations of suspension culture, the fluorescence intensity of the cells had no significant changes, which proved that this site can stably express the EGFP gene. The same method was used to construct recombinant CHO cell lines expressing the human serum albumin (HSA) gene, and was verified by Western blot that this site could express and secrete HSA. It shows that the above-mentioned sites can be integrated and can stably express exogenous proteins.

Objective To preliminary explore the feasibility of three‐dimensional transesophageal echocardiography ( 3D‐T EE) as images data source for 3D printing model by comparing the 3D‐T EE with CT of the aortic root Digital Imaging and Communications in M edicine ( DICOM ) data into 3D printing models respectively . Methods Fifteen patients w ho underwent surgical aortic valve replacement in the hospital were enrolled ,and the aortic root 3D‐T EE and CT DICOM data were obtained in perioperative . T he images were imported into M imics software to generate digital model standard tessellation language file ,and to print the aortic root models by 3D printer . T he structural morphology of both 3D‐T EE and CT models were qualitatively evaluated respectively . T he aortic annular area ,perimeter ,maximal diameter and minimal diameter of the original data , digital model , model and aortic valve replacement were quantitatively evaluated ,and the consistency of each parameter value were analyzed . T he mean diameter of 3D‐T EE and CT model were calculated . T he correlation of mean diameter with the number of replacement was analyzed . Results ①Both 3D‐TEE and CT images data were successfully printed into 3D models ,and the positive rate of aortic valve structure were 93 .3% ( 14/15) and 80 .0% ( 12/15) respectively . ②T he measured values of the aortic annular 3D‐T EE and digital model were smaller than CT ,CTdigital model and replacement ( P＜0 .05) ,and the measurement consistency among groups was high . ③ T he parameter values of 3D‐T EE model were smaller than CT model ( P ＜0 .05 ) ,and the measured values were all within the consistency range . T he mean diameters were highly correlated with the replacement values ( r ＞ 0 .95 , P ＜ 0 .05 ) . Conclusions 3D printing aortic root model based on 3D‐TEE image data is of high feasibility .

Objective: To preliminary explore the feasibility of three-dimensional transesophageal echocardiography (3D-TEE) as images data source for 3D printing model by comparing the 3D-TEE with CT of the aortic root Digital Imaging and Communications in Medicine(DICOM) data into 3D printing models respectively. Methods: Fifteen patients who underwent surgical aortic valve replacement in the hospital were enrolled, and the aortic root 3D-TEE and CT DICOM data were obtained in perioperative. The images were imported into Mimics software to generate digital model standard tessellation language file, and to print the aortic root models by 3D printer. The structural morphology of both 3D-TEE and CT models were qualitatively evaluated respectively. The aortic annular area, perimeter, maximal diameter and minimal diameter of the original data, digital model, model and aortic valve replacement were quantitatively evaluated, and the consistency of each parameter value were analyzed. The mean diameter of 3D-TEE and CT model were calculated. The correlation of mean diameter with the number of replacement was analyzed. Results: ①Both 3D-TEE and CT images data were successfully printed into 3D models, and the positive rate of aortic valve structure were 93.3% (14/15) and 80.0% (12/15) respectively. ②The measured values of the aortic annular 3D-TEE and digital model were smaller than CT, CTdigital model and replacement (P<0.05), and the measurement consistency among groups was high. ③The parameter values of 3D-TEE model were smaller than CT model (P<0.05), and the measured values were all within the consistency range. The mean diameters were highly correlated with the replacement values (r>0.95, P<0.05). Conclusions 3D printing aortic root model based on 3D-TEE image data is of high feasibility.