1.Establishment of a real-time fluorescence quantitative RT-PCR for measurement of t-PA mRNA expression in endothelial cells
Yanting ZHAO ; Lianfen ZHANG ; Jian JIN
Chinese Pharmacological Bulletin 1986;0(05):-
0.990).The intra-assay and inter-assay variation of the method was 3.10 % and 4.93 %,respectively.The all-trans ratinoic acid(ATRA) up-regulated t-PA mRNA expression in a dose-dependent manner(1.25~20.00 ?mol?L~(-1),P
2.Clinical evaluation of plasma D-dimer assay in the diagnosis of deep vein thrombosis of the lower limbs
Chunqi ZHAO ; Weiliang WANG ; Dexiang HUANG ; Yanting HUANG
Chinese Journal of General Surgery 2001;0(09):-
ObjectiveThe aim of this study was to evaluate the measurement of plasma D-dimer level in the diagnosis of acute and chronic deep venous thrombosis(DVT) of the lower limbs. Methods Gold colloids immunofiltration assay(GIA) was used to detect D-dimer in the plasma in 121 patients with DVT of the lower limbs confirmed by continuous wave Dopper ultrasound and B-mode ultrasound (normal value
3.Promotion effect of miR-155 on transforming growth factor β2-induced epithelial-mesenchymal transition in human retinal pigment epithelial cells
Yanting WANG ; Xuemin JIN ; Xiaohua LI ; Zhaoxia ZHAO
Chinese Journal of Experimental Ophthalmology 2021;39(1):13-19
Objective:To investigate the effect of microRNA-155(miR-155) on transforming growth factor β2 (TGF-β2)-induced epithelial-mesenchymal transition of human retinal pigment epithelial cells and its mechanism.Methods:The retinal pigment epithelial cell ARPE-19 cell line was used as the research object.The cells cultured with DMEM medium were served as the control group and the cells cultured with DMEM medium containing 10 ng/ml TGF-β2 were served as the TGF-β2 group.The ARPE-19 cells transfected with miR-155 inhibitor were set as the miR-155 inhibitor group and the ARPE-19 cells transfected with miR-155 negative control were set as the miR-155 negative control group, and the cells in the two groups were cultured in DMEM medium containing 10 ng/ml TGF-β2.After 48 hours cell culture, reverse transcription-PCR was used to detect the expression of miR-155 in each group, and scratch migration test and Transwell chamber test were used to detect cell migration and invasion ability, and Western blot was used to detect the expressions of phosphate and tension homology deleted on chromosome ten gene (PTEN), phosphatidylinositol 3-kinase (PI3K), protein kinase B (Akt), p-Akt and epithelial mesenchymal markers E-cadherin (E-cad), zonula occludens protein 1 (ZO-1), F-actin, α-smooth muscle actin (α-SMA), fibronectin 1 (FN-l) vimentin, proteins.The target gene prediction library predicted miR-155 target gene and fluorescein enzyme reporter vectors were used to identify target genes.Results:After 48 hours of culture, the cells in the control group were in good condition with tight adherence and regular shape.The cells in the TGF-β2 group showed more obvious spindle shape with loose arrangement, and most of the cells were fibrous.The relative expression level of miR-155 in the cells of TGF-β2 group was 0.92±0.14, which was significantly higher than 0.35±0.06 of the control group ( t=7.242, P=0.003). The relative expression level of miR-155 in the cells of miR-155 inhibitor group was 0.21±0.03, which was significantly lower than 0.98±0.09 of the miR-155 negative control group ( t=12.421, P<0.01). The migration rate was higher and the number of cells passing through basement membrane was more in the TGF-β2 group than those of the control group, and the migration rate was higher and the number of cells passing through basement membrane of miR-155 was more in the miR-155 negative control group than those of the miR-155 inhibitor group, and the differences were statistically significant (all at P<0.01). Compared with the control group, the relative expression levels of PTEN, E-cad, ZO-1, F-actin protein were decreased, while the relative expression levels of PI3K and the p-Akt/Akt ratio were increased, and the relative expression levels of α-SMA, FN-1, vimentin proteins were increased in the TGF-β2 group, and the differences were statistically significant (all at P<0.01). Compared with the miR-155 negative control group, the relative expression levels of E-cad, ZO-1, F-actin and PTEN proteins were increased, while the relative expression levels of α-SMA, FN-l, vimentin, PI3K and the p-Akt/Akt ratio were decreased in the miR-155 inhibitor group, and the differences were statistically significant (all at P<0.01). Target gene prediction library prediction and luciferase reporter vector identification confirmed that PTEN was a downstream target gene of miR-155. Conclusions:miR-155 can promote the TGF-β2-induced epithelial-mesenchymal transition progress in human retinal pigment epithelial cells, and its mechanism may be related to inhibiting the expression of the target gene PTEN and stimulating the activation of the PI3K/Akt signaling pathway.
4.Long-term outcomes of carotid artery stenting versus endarterectomy for carotid stenosis: A meta-analysis of randomized controlled trials
Pengfei ZHANG ; Yanting GUO ; Wenke ZHAO ; Liwen ZHAO ; Ziwen WANG ; Yichuan HE ; Yaoyu YU
International Journal of Cerebrovascular Diseases 2017;25(4):310-319
ObjectiveTo evaluate the long-term outcomes of carotid endarterectomy versus carotid artery stenting for carotid stenosis.MethodsPubMed, EMBASE, and the Cochrane databases were retrieved.The randomized controlled trials of comparing CEA with CAS in patients with carotid artery stenosis were enrolled.The data such as the research basic characteristics and the long-term outcomes including stroke or death combined endpoints, any stroke or any death were extracted.The Stata software was used to conduct statistical analysis.ResultsA total of 7 randomized controlled trials and 8 210 patients were included.The median follow-up time was 2-7.4 years.The overall quality of the included studies was high and the risk of bias was low.The meta-analysis showed that the risks of the combined endpoint of stroke or death (hazard risk [HR] 1.21, 95% confidence interval [CI] 1.04-1.39), any stroke (HR 1.32, 95% CI 1.15-1.51) and ipsilateral stroke (HR 1.26, 95% CI 1.02-1.55) in the CAS group were significantly higher than those in the CEA group;the risks of death (HR 1.06, 95% CI 0.95-1.18), disabling stroke (HR 1.23, 95% CI 0.95-1.60), non-ipsilateral stroke (HR 1.12,95% CI 0.81-1.55) and restenosis (HR 1.18,95% CI 0.91-1.52) were not significantly different between between the CAS group and the CEA group.Conclusions CAS and CEA are associated with similar risks of long-term death, disabling stroke, non-ipsilateral stroke and restenosis.The risks of long-term combined endpoint of stroke or death, any stroke and ipsilateral stroke significantly higher with CAS.These results suggest that CEA remains the treatment of choice for carotid stenosis.
5.Application of CT Portography in Esophagogastric Varices in Cirrhotic Patients
Shangfei ZHAO ; Kun FENG ; Qiaoyan QU ; Mingjuan CUI ; Yanting WANG ; Junying TAN ; Mingquan SONG
Chinese Journal of Gastroenterology 2016;21(10):615-619
Background:CT portography(CTP)permits comprehensive evaluation of portal vein and its collateral pathways. It is widely used for assessment of portal hypertension in clinical practice. Aims:To assess the value of CTP in esophagogastric varices in cirrhotic patients. Methods:A retrospective cohort study was performed in 143 cirrhotic patients admitted from Jan. 2013 to Sep. 2015 at the Affiliated Hospital of Qingdao University. All patients fulfilled the inclusion criteria and underwent CTP and gastroscopy within 7 days after admission. According to the occurrence of gastrointestinal bleeding at admission,patients were allocated into two groups:bleeding group( n = 70 ) and non-bleeding group( n = 73 ). Consistency of the results of CTP and gastroscopy was analyzed by kappa coefficient;the accuracy of CTP parameters, including diameters of main portal vein(MPV),splenic vein(SPV)and left gastric vein(LGV)for prediction of variceal bleeding was evaluated by ROC curve. Results:CTP and gastroscopy had a good consistency in typing and grading of esophagogastric varices,with the kappa value of 0. 793 and 0. 775,respectively. The diameters of MPV,SPV and LGV were significantly higher in bleeding group than in non-bleeding group(P < 0. 01),and their area under the ROC curve (AUC)in predicting variceal bleeding was 0. 741,0. 627 and 0. 816,respectively. The accuracy of diameter of LGV was superior to that of MPV and SPV. With the cutoff value of 6. 1 mm,the sensitivity and specificity of diameter of LGV were 65. 71% and 84. 93% ,respectively. With the cutoff value of 16. 3 mm,the sensitivity of diameter of MPV was 75. 71% , which was higher than that of LGV and SPV. Conclusions:CTP can be used in clinical diagnosis of esophagogastric varices in cirrhotic patients,and two CTP parameters,the diameters of LGV and MPV,might be helpful for prediction of variceal bleeding.
6.The role of human fibroblast growth factor receptor 1-Ⅲb isoform in pancreatic cancer cells
Zhanbing LIU ; Yinmo YANG ; Qilu QIAO ; Jianxun ZHAO ; Yanting HUANG ; Marko Kornmann
Medical Journal of Chinese People's Liberation Army 2007;32(2):88-91
Objective To study the role of Ⅲb isoform of human fibroblast growth factor receptor 1 (FGFR1-Ⅲb) in PANC-1 pancreatic cancer cells. Methods The plasmid of human full-length FGFR1-Ⅲb isoform,pSVK4/FGFR1-Ⅲb, was stable transfected into cultured PNAC-1 pancreatic cancer cell lines facilitated by lipofectamine. The function of FGFR1-Ⅲb in transfected pancreatic cancer cells were examined by MTT assay, soft agar assay, cell migration assay, single cell movement assay, In vivo tumorigenicity assay. Results The basal anchorage-dependent and -independent cell growth was significantly inhibited. Additionally, FGFR1-Ⅲb expression inhibited single cell movement and in vitro invasion as determined by time-lapse microscopy and boyden chamber assay as well as in vivo tumor formation and growth in nude mice. Microscopic analysis of the xenograft tumors revealed a reduced Ki-67 labelling, lower amount of tumor necrosis and higher grade of differentiation in FGFR1-Ⅲb expressing tumors. Conclusion We identified a functional human FGFR mRNA splice variant that inhibits the transforming potential of pancreatic cancer cells.
7.Effect of different doses of propofol on cisplatin-induced hepatotoxicity in rats
Limei ZHAO ; Renchun LAI ; Jing WANG ; Yanting FAN ; Qin WANG ; Xudong WANG
Chinese Journal of Anesthesiology 2012;(10):1186-1188
Objective To investigate the effect of different doses of propofol on cisplatin-induced hepatotoxicity in rats.Methods Eighty male SD rats weighing 200-250 g,aged 3 months,were randomly divided into 8 groups (n =10 each) ∶ control group (group C),cisplatin 7.5 mg/kg group (group Cis),propofol 180 mg/kg group (group P),intralipid 15 ml/kg group (group Ⅰ),cisplatin 7.5 mg/kg + intralipid 15 ml/kg group (group CisI),cisplatin 7.5 mg/kg + propofol 60 mg/kg group (group CisP1),cisplatin 7.5 mg/kg + propofol 120 mg/kg group (group CisP2),and cisplatin 7.5 mg/kg + propofol 180 mg/kg group (group CisP3).The rats in groups C,Cis,P and I received single intraperitoneal injection of normal saline,cisplatin,propofol and intralipid respectively.While in the groups CisI,CisP1,CisP2 and CisP3,rats received a single intraperitoneal injection of propofol or intralipid at 1 min before a single intraperitoneal injection of cisplatin.At 24 h after cisplatin injection,venous blood samples were taken from inferior vena cava for measurement of plasma ALT and AST activities.The liver tissues were taken for microscopic examination.Results Compared with group C,plasma ALT and AST activities were significantly increased and pathological injury was aggravated in groups Cis,CisI and CisP1-3 (P < 0.05).The activities of ALT and AST were gradually decreased and the pathologic injury was attenuated in groups Cis and CisP1-3 (P < 0.05).Conclusion Propofol can reduce cisplatin-induced hepatotoxicity in a dose-dependent manner in rats.
8.Effect of propofol on docetaxel-induced toxicity to cervical cancer Hela cells transfected with Cx32 plasmid
Yanting FAN ; Renchun LAI ; Jing WANG ; Limei ZHAO ; Qin WANG ; Xudong WANG
Chinese Journal of Anesthesiology 2013;33(6):687-690
Objective To evaluate the effect of propofol on docetaxel-induced toxicity to cervical cancer Hela cells transfected with Cx32 plasmid.Methods Cervical cancer Hela cells transfected with Cx32 plasmid were seeded at two different densities and induced to express Cx32 by incubation with doxycycline for 48 h.The cells at high density were seeded at 1 × 105 cells/ml such that the cells would be confluent at the time of docetaxel exposure.The cells at low density were seeded at 500 cells/ml and the cells did not attach at the density.Each type of cells obtained was randomly divided into 5 groups (n =8 each):control group (group C),docetaxel group (group D),docetaxel + intralipid group (group D + I),docetaxel + 18-α-GA group (D + 18-α-GA),and docetaxel +propofol group (group D + P).Groups D,D + I,D + 18-α-GA and D + P were exposed to 5 nmol/L docetaxel,5 nmol/L docetaxel + 10μg/ml intralipid,5 nmol/L docetaxel + 10 μmol/L 18-α-GA,and 5 nmol/L docetaxel +2.8 μg/ml propofol,respectively.18-α-GA,intralipid and propofol were added prior to docetaxel,and the action time for 18-α-GA alone was 1 h and for intralipid or propofol alone 2 h.The time for coaction between the three drugs and docetaxel was 2 h.Cell survival was determined by a standard colony-forming assay.Results The colony formation rate of the cells seeded at high density in group D was significantly lower than that of the cells seeded at low density in group D (P < 0.05).For the cells seeded at high density,the colony formation rate was significantly decreased in the other groups when compared with group C (P < 0.05).The colony formation rate was significantly higher in groups D + 18-α-GA and D + P than in groups D and D + I (P < 0.05).There was no significant difference in the colony formation rate between groups D and D + I (P > 0.05).For the cells seeded at low density,the colony formation rate was significantly decreased in the other groups when compared with group C (P < 0.05) and there was no significant difference in the colony formation rate between D,D + I,D + 18-α-GA and D + P groups (P > 0.05).Conclusion Propofol can attenuate docetaxel-induced toxicity to Hela cells transfected with Cx32 plasmid and inhibition of gap junction function is involved in the mechanism.
9.The negative effects of Trim34α on activation of luciferase reporter gene containing NF-κB promoter
Dakang SUN ; Zhenguo SU ; Xinye AN ; Rongjiao ZHOU ; Xiangqin SONG ; Yanting ZHAO
Chinese Journal of Microbiology and Immunology 2011;31(6):492-497
Objective To investigate the effects of Trim34α on the activation of luciferase reporter gene containing NF-κB promoter induced by adaptor proteins TAB2. Methods The total RNA was isolated from HeLa cells. After amplification with RT-PCR, the target sequences were cloned into 5'-Flag-pcDNA3.1 (+) vector. The recombinant vector was confirmed by restriction enzyme digestion, colony PCR and sequencing. It was transfected into HEK293T cells to detected Trim34α expression by Western blot. Simultaneously, the effects of Trim34α on the NF-κB activation induced by TAB2 were determined by dual-luciferase reporter assay. Results Restriction enzyme digestion, colony PCR and sequencing confirmed the vector was constructed successfully, furthermore it expressed Trim34α protein in HEK293T cells. Moreover, trim34α could form high-molecular-weight oligomeric protein, and here we called it trimsome. Interestingly, dual-luciferase assay showed that Trim34α could effectively block TAB2-induced NF-κB activation. Conclusion Trim34α was involved in negative regulation of TAB2-induced NF-κB activation and could form high-molecular-weight oligomer.
10.One phase treatment of renal calculi accompanied with pyonephrosis by percutaneous nephrolithotripsy
Xianming FAN ; Jianxing LI ; Yongqiang XIA ; Jing NIU ; Kang SUN ; Pengfei WANG ; Yanting ZHAO
Chinese Journal of Urology 2008;29(9):621-623
Objective To evaluate the method, clinical efficacy and safety of one phase treat-ment of renal calculi associated with pyonephrosis by percutaneous nephrolithotripsy(PCNL) by pneu-matic combined with ultrasonic lithotriptor. Methods Sixty-six cases of renal calculi accompanied with pyonephrosis were treated with PCNL. The renal calyx was punctured under ultrasound gui-dance, then the tract was dilated from F8 to F16 by peel-away vascular access sheathes. After the in-sertion of the flexible sheath, metallic dilator was inserted and the flexible sheath was pulled out. The tract was dilated by metallic sheath to F21 and the operation sheath and nephroseope were placed into working tract. EMS III LithoClast Master was used. Ultrasonic powered lithotriptor probe with suc-tion was used to clear the liquor puris and calculus fragments with low-pressure or no-pressure. The combined pneumatic and ultrasonic powered lithotriptor was used to break and clear the calculi. Re-salts Of the 66 cases, there was no bacteremia or pyaemia intraoperatively and postoperatively. And there was no other severe complication occurred intraoperatively. One phase PCNL was successfully completed in 60 cases. Other 4 cases had residual calculi less than 1.5 em in diameter and received ESWL to break the calculi, 2 cases had bigger residual calculi and accepted second PCNL 1 week after the first intervention. In the follow-up period, the 3 month post-operative serum Cr was 56-203 μmol/L with an average decrease of 40 μmol/L, GFR was 5.0-56.2 ml/min with an average increase of 23.6 ml/min compared with the pre-operative data. At 6 months postoperative serum Cr was 56-158 μmol/L with average decrease of 31 μmol/L, GFR was 5.0-79.2 ml/min with an average in-crease of 30.2 ml/min. Conclusion Application of PCNL in the treatment of patients with renal cal-culi accompanied with pyonephrosis is safe, cost-effective and clinically efficient by pneumatic com-bined with ultrasonic lithotriptor.