Objective To purify recombinant methioninase and investigate the synergetic inhibitory effect of methioninase and cisplatin on the proliferation of human lung adenocarcinoma cell line GLC. Methods Recombinant methioninase was purified with GST-column from supernatant after ultrasonic disruption of cultured Escherichia coli in the prokaryotic expression system pGEX-4T-1-Met/Dh5a. MTT assay was used to determine the inhibition rate of methioninase in combined with cisplatin on cell proliferation,and their synergistic effect was evaluated by using the q value judge method. Results The concentration of recombinant methionine cleaving enzyme was 0.22 mg/mL, the purity was 95%, and the activity was 0.568 IU/mg. After 72 h culturing, the inhibition rate of cisplatin methionine was 24.80%and 27.49%respectively,while the inhibition rate of the combined drugs was 66.80% ( =1.47>1.15) which showed a significant synergistic effect. Conclusion Both methioninase and cisplatin have the inhibition effect,and the combined drugs display a significant synergistic effect on the proliferation of GLC.

Objective:To investigate whether hsa_circ_0054595(circRTN4)is involved in glomerulosclerosis of lupus nephri-tis(LN)by regulating monocyte-derived TNF-α.Methods:RT-qPCR and immunofluorescence were used to detect the expression of TNF-α in monocytes of LN patients,and fluorescence in situ hybridization(FISH)was used to detect the expression of circRTN4.THP1 cells were transfected with circRTN4-siRNA and negative control NC-siRNA,respectively.RT-qPCR and Western blot were used to detect the expression of TNF-α in THP1cell,and ELISA was used to detect the expression of TNF-α in THP1 culture superna-tant.THP1 cells were transfected with mircoRNA mimics,and the expression of TNF-α was detected by Western blot.The direct bind-ing of miR-486-3p to TNF-α and circRTN4 were verified by reversion experiment and dual luciferase reporter gene experiment.MRL/lpr mice were injected with circRTN4 recombinant adeno-associated virus via tail vein,and the expression of circRTN4 in peripheral blood mononuclear cells of the mice was detected by RT-qPCR.ELISA was used to detect the expression of serum TNF-α.HE staining and PAS staining were used to observe the pathological changes,and immunofluorescence was used to detect the expression of FN.Results:The expressions of TNF-α and circRTN4 were increased in monocytes of LN patients(P<0.05).The expression of TNF-α in THP1 was significantly increased in the LN group(P<0.01).Knockdown of circRTN4 inhibited the expression and secretion of TNF-α(P<0.05),and this effect was achieved by binding to miR-486-3p(P<0.01).In vivo,knockdown of circRTN4 in peripheral blood monocytes of MRL/lpr mice reduced the expression of TNF-α in serum(P<0.05),and improved glomerular cell proliferation and FN deposition.Conclusion:Highly expressed circRTN4 in monocytes promotes the expression of TNF-α by binding to miR-486-3p,and participates in the occurrence and development of glomerulosclerosis in LN.

Objective:To analyze the relevant risk factors for restenosis after percutaneous transluminal angioplasty of patients with Takayasu′s arteritis.Methods:Clinical data of 43 patients undergoing percutaneous angioplasty due to Takayasu arteritis were retrospectively analyzed. Univariate and multivariate Logistic regression analysis was used to explore the relevant risk factors for restenosis after percutaneous transluminal angioplasty.Results:There were 9 males and 34 females. The mean age was 23 (18-33) years old, 59 times of PTA were performed, including 44 in renal artery, 9 in aorta, 2 in iliac and 2 in carotid artery, 1 in brachiocephalic trunk and 1 in left subclavian artery. The mean follow up time was (64±42) months. The rate of restenosis was 47.5%(28/59)and the mean time of restenosis was (23±27) months. The restenosis rate of aorta and iliac artery was 9.1%, that of renal artery was 52.3% and that of supra aortic artery was 100% . The rate of restenosis was higher in patients with symptoms of headache, syncope and low back pain, the elevated ESR and CRP increased the risk of restenosis (all P<0.05). Multivariate Logistic analysis showed that preoperative elevation of ESR and CRP were risk factors for restenosis after percutaneous angioplasty for Takayasu arteritis. Conclusions:PTA was safe and effective in Takayasu arteritis involving aorta-iliac and renal artery, the elevated ESR and CRP was related to high risk of restenosis.