1.Down-regulation of ubiquitin specific peptidase 18 suppresses the invasion and proliferation of gastric cancer cells
Xianfei ZHONG ; Lizhi YI ; Xuejie DENG ; Zhengyu CHENG ; Qin WANG ; Jianmei WANG
Cancer Research and Clinic 2018;30(7):433-437
Objective To analyze the expression and role of ubiquitin specific peptidase 18 (USP18) in gastric cancer cells,and to investigate the relationship between the development of gastric cancer and USP18.Methods The levels of USP18 protein and mRNA expression in immortalized gastric mucosa epithelial GSE cell lines and gastric cancer cell lines (AGS,MKN45,MKN25,BGC823,BGC803,SGC7901) were detected by using reverse transcription-polymerase chain reaction (RT-PCR) and Western blot,respectively.The role of USP18 in the invasion and proliferation of gastric cancer cells was analyzed by using CCK8 and Transwell assays.Results The mRNA level of USP18 was lower in GSE cell lines than that in gastric cancer cells (F =794.052,P < 0.000 1).In six gastric cancer cell lines,mRNA level of USP18 was relatively high in BGC823 (17.62±0.55) and BGC803 (13.52±0.50) cell lines,and low in MKN28 (1.40±0.17) and MKN45 cell lines (4.23±0.26).As for the protein level,the expression of USP18 was lowest in GSE cell line.In six gastric cancer cell lines,the expression of USP18 was the highest in more aggressive SGC7901 and BGC803 cell lines and the lowest in AGS and MKN45 cells.Compared with the control group,interference of USP18 decreased the invasion and proliferation abilities of SGC7901 and BGC803 cell lines (P < 0.01).Conclusion USP18 is highly expressed in more invasive gastric cancer cells,and the downregulation of USP18 can suppress the invasion and proliferation of gastric cancer cells.
2.Effect of apoptosis-inducing factor gene knockdown on bone marrow mesenchymal stem cell transplantation for myocardial infarction
Dunzheng HAN ; Xiaozhou QIN ; Xiudi PAN ; Waner LU ; Ying DAI ; Yanxun CHEN ; Xianfei CHENG ; Muhan TANG
Chinese Journal of Tissue Engineering Research 2024;28(25):3967-3973
BACKGROUND:Numerous basic and clinical trials have confirmed that the low survival rate after bone marrow mesenchymal stem cell transplantation is a serious constraint on its long-term therapeutic effect.Previous studies have shown that apoptosis-related factors play an important role in the apoptosis of bone marrow mesenchymal stem cells,of which apoptosis-inducing factor may be a key factor. OBJECTIVE:Bone marrow mesenchymal stem cells,of which apoptosis-inducing factor was knocked down,were transplanted into infarcted myocardium of mice,aiming to certify the importance of apoptosis-inducing factor in the survival of bone marrow mesenchymal stem cells to further recover cardiac function after infarction. METHODS:Firstly,bone marrow mesenchymal stem cells were infected with LV-AIF-shRNA lentivirus to down-regulate the expression of apoptosis-inducing factor protein.Flow cytometry,western blot assay,and RT-qPCR were used to detect the infection efficiency of lentivirus.CCK-8 assay was used to detect the cell viability of bone marrow mesenchymal stem cells with apoptosis-inducing factor knockdown under hypoxic and ischemic conditions.Then,with the mouse model of acute myocardial infarction constructed,the normal bone marrow mesenchymal stem cells and bone marrow mesenchymal stem cells with apoptosis-inducing factor gene knockdown were transplanted into the model,respectively.The expression of apoptosis-inducing factor was examined by fluorescence immunoassay.Serum brain natriuretic peptide levels were detected by ELISA.Cardiac ultrasound was used to detect cardiac function.Myocardial fibrosis was observed by Masson staining.The expression of SRY gene was detected by RT-qPCR in apoptosis-inducing factor-knocked bone marrow mesenchymal stem cells after transplantation,reflecting cell survival. RESULTS AND CONCLUSION:(1)Bone marrow mesenchymal stem cells with apoptosis-inducing factor gene knockdown were successfully established by LV-AIF-shRNA lentivirus infection,following 97.7%of infection efficiency,and notably decline of the expression of apoptosis-inducing factor(P<0.001).(2)Under ischemia and hypoxia,the cell viability of apoptosis-inducing factor knockdown bone marrow mesenchymal stem cells was significantly increased compared with normal bone marrow mesenchymal stem cells.(3)Compared with normal bone marrow mesenchymal stem cells after transplantation,the survival number of bone marrow mesenchymal stem cells in the infarcted myocardium after apoptosis-inducing factor gene knockdown was significantly increased to 3.71 times(P<0.001),and the apoptosis-inducing factor protein expression and myocardial fibrosis degree in the infarcted area were significantly reduced.(4)Compared with normal bone marrow mesenchymal stem cells,the serum brain natriuretic peptide level of bone marrow stem cells with apoptosis-inducing factor gene knockdown after transplantation was significantly decreased(P<0.05),and left ventricular ejection fraction and left ventricular shortening fraction were significantly improved(P<0.05).(5)These findings confirm that apoptosis-inducing factor gene knockdown can reduce myocardial fibrosis and improve cardiac function after acute myocardial infarction via enhancing the bone marrow mesenchymal stem cell viability and increasing the bone marrow mesenchymal stem cell survival after transplantation in the donor.
3.Study on the relationship between cystatin C level and the plasma trough concentration of teicoplanin
Qing DAI ; Lin CHENG ; Xianfei HU ; Fang LIU
Journal of Pharmaceutical Practice 2021;39(6):569-572
Objective To investigate the relationship between cystatin C level and the plasma trough concentration of teicoplanin, so as to provide a reference for the rational application of teicoplanin in clinical practice. Methods The clinical data of the patients receiving teicoplanin, who admitted to our hospital from October 2017 to July 2020 were retrospectively analyzed. The distribution of teicoplanin concentration, the difference of teicoplanin concentration under different cystatin C level, and influence factors for teicoplanin concentration (<15 µg/ml) were analyzed. Results A total of 98 patients including 65 males and 33 females, aged 19 to 94 (52.2±16.2) years old, with 141 trough concentrations were enrolled. The trough concentration of teicoplanin was 11.51 (8.35, 19.07) µg/ml, and the range was 3.57-41.93 µg/ml. 95 cases (67.38%) had teicoplanin concentration <15 µg/ml. When the concentration of cystatin C was >1.05 mg/L, the trough concentration of teicoplanin were 11.37 (8.96, 20.52) µg/ml, significantly higher than those when the concentration of cystatin C was in normal [8.68 (6.34, 11.79) µg/ml, Z=−2.636, P<0.05]. Logistic regression analysis showed that cystatin C level was the influencing factor for teicoplanin trough concentration does not meet the standard (OR=1.529, 95%CI=1.001-2.336, P<0.05). Conclusion The concentration of teicoplanin is significantly increased when the cystatin C level is higher than the normal. Cystatin C level is the influence factor for teicoplanin trough concentration not meeting the standard. The cystatin C level may be considered as a reference for teicoplanin dosage adjustment in clinical practice.