Objective To investigate the expression of recombinant IL-37b protein and removal of the endotoxin, and identify its biological activity.Methods The prokaryotic expression vector pET28/IL-37b was constructed and to transform Escherichia coli (E.coli) Rosetta.After induction with IPTG, the recombinant protein was purified through Ni2+-NTA gel column and identified by SDS-PAGE and Coomassie brilliant blue staining.Then, the endotoxin protein was removed and was treated with LPS-stimulated RAW 264.7 cells.The culture supernatant was collected.The expression of IL-6 was detected by ELISA and the biological activity of the protein was identified.Results The recombinant IL-37b with high purity was expressed and the endotoxin produced by prokaryotic expression was reduced, and it was identified to have good biological activity.Conclusions In this study a recombinant IL-37b protein with high biological activity is successfully obtained.

Inteleukin-37 is a member of IL-1 family and originally named as IL-1F7.It has five different subtypes (IL-37a-e).It has been reported that IL-37 suppresses pro-inflammatory cytokines production of a variety of immune cells, and further regulate innate immune response. In addition, IL-37 has protective effects on colitis, arthritis, pancreatitis and other inflammatory diseases that induced by exogenous stimuli. In a word, IL-37 is a novel anti-inflammatory cytokine and plays an important role in a variety of inflammation-related diseases.

Objective To establish lentiviral expression vectors for Smurf1 silencing and assess the effects of Smurf1 silencing on cell migration.Methods HeLa and A549 cells were infected with lentiviral expression vectors for Smurf1 silencing respectively.After 7 days,the stable cell lines with Smurf1 silencing were obtained after puromycin-resistance screening,enrichment and expansion.The intracellular gene and protein levels of Smurf1 were detected by qPCR and western blot.Transwell assay was used to assess the effect of Smurf1 silencing on cell migration.Results The stable cell lines with Smurf1 silencing are constructed successfully.Silencing of Smurf1 down-regulated cell migration rate detected by Transwell assay.Conclusion Smurf1 promotes cell migration.

Objective To study the feasibility and long-term effects in 173 infants with congenital ventricular septal defect who underwent transcatheter therapy by double-disk ventricular septal defect occluder. Methods We analyzed the clinical data of 173 infants with ventricular septal defect who were performed interventional therapy followed by a follow up study from December 2002 to October 2013. Results The procedure was performed in 173 infants (male = 81 cases and female = 92 cases) aged 11 month-3 years[(2.1±0.7)years]. The weight were 6-15 kg[(10.2±3.6)kg]. The diameter of the defects ranged from 2.5-9.0 mm[(5.1±1.7) mm]. The characteristics of septal were classiifed into four types:the simply perimembranous ventricular septal defects (88 patient,50.8%), the perimembranous ventricular septal defects with pseudoaneurysm (52 patients, 30.0%), the perimembranous ventricular septal defects complicated with aortic valve prolapse (13 patients, 7.5%), and the intracristal ventricular septal defect (20 patients, 11.7%). The diameter of the occluders were between 4-12 mm[(6.3±2.2)mm]. The period of follow-up ranged from 1 month-10 years[(6.2±1.3)years]. The 168 defects were completely occluded in 173 patients(97.1%closure rate) except 5 infants, which 3 patients because the occluder hinder the function of aortic valves and the other occurred complete atrioventricular block(CAVB) when the catheter through defect. One patient occurred CAVB on the third day after the procedure, was reversed by dexamethasone after 4 days. 3 patients with complete left bundle branch block(CLBBB) after the procedure and one went to sustained, but did not observed heart failure in this case during 5 years follow-up. One patient observed sustained CLBBB on the fourth year after the procedure, also did not observed heart failure in this case during 3 years follow-up. 8 patients with trivial residual shun and 4 patients with newly appearance of trivial aortic regurgitation after the procedure, the shun all disappear at one year and the regurgitation did not progress during longest follow-up period at 8.5 years. No other complications, including late-onset CAVB, hematolysis, the occluder displacement and detachment, occurred during 1 month-10 years of follow-up. Conclusions Transcatheter closure of congenital ventricular septal defects is an efifcient method that can be safely used in the majority of infants with ventricular septal defects and have signiifcant long-term effects.

Objective To investigate the effect of Smad3 on cell migration of A549 and HeLa cells.Methods Primers for pCMV-Myc-Smad3 plasmid construction and siRNA targeting Smad 3 were designed and synthesized .pCMV-Myc-Smad3 plasmid was constructed with molecular cloning techniques .Overexpression of Smad 3 with Myc-tag or silencing of endogenous Smad3, and then scratch assay was used to detect the migration ability of A 549 and HeLa cells in vitro. Results pCMV-Myc-Smad3 plasmid was successfully constructed .Overexpression of Smad 3 significantly up-regulated the migration rate of A549 and HeLa cells.Conversely, in the same cells, silencing of endogenous Smad3 or treatment with Smad3 inhibitor, SIS3, down-regulated the migration rate .Conclusions Smad3 promotes cell migration of A549 and HeLa cells.

Objective To establish a patient-derived xenografts (PDX) mouse model of liver cancer (LC) and to explore its role in precision medicine.Methods PDX model was established by subcutaneous implantation of tumor tissues in NCG mice.The morphological structure of tumor tissue was exaimed using HE staining.Fifteen BALB/c nude mice were subcutaneously inoculated with tumor cell suspension from the PDX models.The xenograft mice were randomly divided into 5-fluorouracil (5-FU) group, sorafenib group and negative control group.The tumor volume and body weight of the tumor-bearing mice were measured regularly, the tumor inhibition rate was calculated and the curative effect was evaluated.Results The success rate was 33.3% (6/18) in the establishment of liver cancer PDX mouse model, and the model well retained the characteristics of the primary tumor.In one case of PDX mouse model, the tumor inhibition rates of 5-FU and sorafenib group were 63.7% and 29.6%, with a statistically significant differece between them (P< 0.05), and there was no significant difference between the sorafenib group and negative control group, consistent with clinical observation.Conclusions The PDX mouse model of liver cancer can maintain the histological structure of primary tumor, and can be applied to precision medicine for patients with liver cancer.

Objective To construct a eukaryotic expression vector pEGFP N1/IL-37b of full-length and mature IL-37b,and to detect the expression of both full-length and mature IL-37b in RAW 264.7 cells, a mouse macrophage cell line. Methods To construct the eukaryotic vectors of full-length and mature IL-37b by using plasmid pUBC/IL-37b as a template containing the coding region of IL-37b full-length gene. To detect the expression of IL-37b by western blot and confocal microscopy after transfected the recombinant plasmid into RAW 264.7 cells, and to detect the inhibition of full-length and mature IL-37b on IL-6 production by real-time PCR. Results Eukaryotic vectors pEGFP N1/IL-37b expressed full-length and mature IL-37b after transfection in cells, which inhibited LPS-induced IL-6 production. Conclusions Eukaryotic vectors of full-length and mature IL-37b can be successfully constructed,and lays a foundation for further study of anti-inflammation functions and mechanisms of IL-37b.