0.05).The differences between NSCLC and SCLC reached the statistical signifcance,P

Objective: To investigate the expression of recombinant human intedeukin-18 (hIL-18) in 3.7 L fermenter with the constructed engineer train Pichia pastoris X-33/hIL-18, and the procedures of expression and purification thereof.Methods: The train X-33/hIL-18 was inoculated in BMGY medium and then inoculated into the fermenter until the A600 of the culture reached about 6. The supernatant of fermentation was isolated and purified with centrifugal fiher devices, hydrophobic chromatography column and anion exchange chromatography column. Results: The recombinant hIL-18 was expressed in 3.7 L fermenter with batch feed methanol and the concentration reached 202 mg/L. After the purification, the purity could be more than 97%. The recombinant hIL-18 was shown to induce interferon-gamma (IFN-γ) production by human peripheral blood mononuclear cells (PBMCs), and enhanced NK cell cytotoxicity synergistically with IL-2. Conclusion: A great deal of the recombinant hIL-18 with higher purity could be harvested by Pichiapastoris expression system. This study showed a new potential for further study of its function and activities.

To study the effect of sphingosine-1-phosphate (S1P) on the cardiomyogenic differentiation of human umbilical cord mesenchymal stem cells (UC-MSCs) and human adipose-derived mesenchymal stem cells (AD-MSCs), we seeded the cells in the culture plates and used cardiomyocyte culture medium (CMCM) combining with different concentration of S1P to induce UC-MSCs and AD-MSCs in vitro for 7, 14 and 28 days. Cardiomyogenic differentiations were identified through immunofluorescence staining, and the results were observed with fluorescence microscopy and confocal microscopy. The effects of S1P and CMCM on cell activity were evaluated by the methyl thiazolyl tetrazolium assay. The functional characteristic similar to cardiomyocytes was evaluated through detecting calcium transient. Our results showed that cardiomyogenic differentiation of UC-MSCs or AD-MSCs were enhanced with S1P concentration increasing, but cell activities declined. Results showed that the suitable differentiation time was 14 days, and the optimal concentration of S1P was 0.5 micromol/L. When working together with CMCM, S1P could promote the differentiation of UC-MSCs or AD-MSCs into functional cardiomyocytes, giving rise to specific electrophysiological properties (the calcium transient). Taken together, our results suggested that S1P could promote the differentiation of UC-MSCs or AD-MSCs into functional cardiomyocytes when being cultured in CMCM.
Adipose Tissue ; cytology ; metabolism ; Cell Differentiation ; drug effects ; Cells, Cultured ; Culture Media ; Humans ; Lysophospholipids ; pharmacology ; Mesenchymal Stromal Cells ; cytology ; drug effects ; Myocytes, Cardiac ; cytology ; Sphingosine ; analogs & derivatives ; pharmacology ; Umbilical Cord ; cytology

Objective: To examine the relationship between clustering of metabolic disorders and left ventricular hypertrophy (LVH) in hypertensive children and adolescents, and to provide the reference for preventing the damage to their heart structure. Methods: Based on the data from the Twelfth Five-year National Science and Technology Support Program “Early Warning, Diagnosis and Treatment of Cardiovascular Disease in Children” from September 2012 to September 2014 in Jinan, 261 children and adolescents aged 6-17 years whose hypertensive status was confirmed based on appearance of elevated blood pressure across three different occasions were included. Covariance analysis was used to analyze the relationship of clustering of metabolic disorders with left ventricular mass index levels. Multivariable logistic regression model was used to examine the relationship of clustering of metabolic disorders with LVH. Results: After adjustment for sex, age and blood pressure, LVMI levels were (33.21±1.85) (38.57±1.19) and (43.00±1.63)g/m2.7, respectively, for hypertensive children and adolescents carrying 0, 1 and ≥2 metabolic disorders; LVMI levels increased with the number of metabolic disorders in hypertensive children and adolescents (P<0.01). Compared with the hypertensive children and adolescents without any metabolic disorders, the risk for LVH increased in ones carrying 1 and ≥2 metabolic disorders, the odds ratios and 95% confidence intervals were 2.41(1.11-5.23) and 4.69(2.05-10.74), respectively. Conclusion The clustering of metabolic disorders was positively correlated with the LVMI levels and risk of LVH in hypertensive children and adolescents. Therefore, to prevent cardiac damage such as LVH in hypertensive children and adolescents, it is important to prevent and control metabolic disorders comprehensively.

Objective To investigate the preventive effect of specially-designed clothing on dressing the wound of panents after breast cancer surgery. Methods Using random number table, we divided 400 patients having undergone breast cancer surgery into the observation group and control group in equal number. In the control group, the patients wore ordinary patient clothing after dressing the chest wound with compression bandage,and in the observation group the patients wore the specially-designed clothing after the same treatment of the chest wound as that in the control.The two groups were compared in terms of the incidence of subcutaneous effusion,incidence of skin flap necrosis and the difference of patients'comfort. Results The incidences of subcutaneous effusion and skin flap necrosis were significantly lower than those in the control group.The patient was more comfortable than that the control group, with statistically significant differences (P<0.05,P<0.01). Conclusion After breast cancer surgery,the specially-designed clothing for the patients can effectively reduce the incidences of subcutaneous effusion and incidence of skin flap necrosis and improve the patient's comfort,and deserve clinical promotion.

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.

Objective To investigate the effect of dynamic monitoring of occlusal force on the final therapeutic effect and the change of periodontal supporting tissue during combined periodontal orthodontic treatment.Methods The periodontal clinical index of 20 patients with traditional periodontal orthodontic treatment and 20 patients with combined periodontal orthodontic treatment assisted by T-Scan Ⅲ and Anycheck digital occlusion analysis system were compared before,during and after treatment,as well as the changes of bite force,bite time and tooth mobility in the experimental group.Results The depth of periodontal pocket(PD),loss of attachment(AL),bleeding index(BI)and tooth looseness were significantly reduced after combined periodontal orthodontic treatment in both groups.In the control group,the percentage of anterior and posterior biting force changed obviously,and the occlusion force balance was improved.Conclusion The combined treatment of periodontitis and orthodontics can improve the periodontal tissue of patients with periodontitis,and T-Scan system can observe and guide the adjustment of occlusal and better achieve occlusion force balance.