1.Effect of propofol preconditioning on endoplasmic reticulum stress induced by hypoxia-reoxygenation in HEPG2 cells
Xiuli YANG ; Weixiang TANG ; Likui WANG ; Yujun SHEN ; Yuanhai LI
Chinese Journal of Anesthesiology 2014;34(4):489-491
Objective To evaluate the effect of propofol preconditioning on endoplasmic reticulum stress induced by hypoxia-reoxygenation (H/R) in HEPG2 cells.Methods HEPG2 cells were randomly divided into 4 groups using a random number table:control group (group C),propofol group (group P),H/R group and H/R + propofol preconditioning group (group PP).In group C,the cells were cultured routinely for 42 h.In group H/R,after being cultured routinely for 6 h,the cells were exposed to 1% O2 + 5% CO2 + 94% N2 for 12 h followed by 12 h reoxygenation.In group PP,the cells were cultured for 6 h in the culture medium containing propofol 10 μmol/L (final concentration),and then H/R was induced.The cell viability was detected by MTT assay.The expression of immunoglobulin heavy chain-binding protein (BIP),C/EBP homologous protein (CHOP) and activated caspase-3 was determined by Western blot.The expression of BIP,CHOP and caspase-3 mRNA was determined by RT-PCR.Results Compared with group C,the cell viability was significantly decreased,and the expression of BIP,CHOP and activated caspase-3 protein and mRNA was up-regnlated in H/R and PP groups,and no significant changes were found in the parameters mentioned above in group P.Compared with group H/R,the cell viability was significantly increased,and the expression of BIP,CHOP and activated caspase-3 protein and mRNA was down-regulated in PP group.Conclusion Propofol preconditioning can promote the cell proliferation and attenuate H/R injury to HEPG2 cells through inhibiting endoplasmic reticulum stress.
2.Characterization of Cd and Cu Complexing with Metallothionein by Eletrospray Ionization Mass Spectrometry
Jincan SHEN ; Weixiang YE ; Haining KANG ; Liya GE ; Zhixia ZHUANG ; Xiaoru WANG
Chinese Journal of Analytical Chemistry 2009;37(7):975-979
Mammal metallothionein(MT) folds into two separate domains that exhibit different structure and metal binding propertity independently, the study of the strategy of metal ions binding with MT would give better understanding of their exact biological functional mechanisms. In this study, a method using eletrospray ionization mass spectrometry (ESI-MS) phase liquid chromatography and identified by ESI-MS. Different amounts of Cd or Cu were then added in MT-2a samples and ESI-MS was employed to determine the mass difference of MT in different samples. The results Cd2+4S11; while Cd is attached in separate binding sites for the formation of Cd2+3S9 cluster, which intermediately formed with five and six Cd ions were detected. For the Cuprous ions, it prefers to cooperatively bind in β-domain with the form of Cu4-MTβ. The binding form in β-domain would convert from Cu4 into more Cu binding form with the addition of Cu. When high concentration of Cu was added in samples, the result suggested that
3.Effect of propofol pretreatment on endoplasmic reticulum stress-mediated apoptosis during focal cerebral ischemia-reperfusion in rats
Weixiang TANG ; Likui WANG ; Yiqiao WANG ; Xiuli YANG ; Zhixin GAO ; Yujun SHEN ; Yuanhai LI
Chinese Journal of Anesthesiology 2012;(12):1488-1490
Objective To evaluate the effect of propofol on the endoplasmic reticulum stress-mediated apoptosis during focal cerebral ischemia-reperfusion (I/R) in rats.Methods Eighty adult male Sprague-Dawley rats,weighing 240-280 g,were randomly divided into 4 groups (n =20 each):shame operation group (group S) ; focal cerebral I/R group (group FCIR); propofol pretreatment group (group P); intralipid pretreatment group (group Ⅰ).Focal cerebral I/R was induced by 4 h middle cerebral artery occlusion followed by reperfusion.Propofol was infused at a rate of 12 mg· kg-1 · h-1 starting from 30 min before ischemia until 15 min of ischemia in group P,while intralipid was given instead of propofol in group I.Neurological deficit scores (NDSs) were measured at 6 h of reperfusion in 10 rats chosen from each group and the rats were then sacrificed.Their left brains were removed for determination of brain water content.The left 10 rats were sacrificed and their brains were immediately removed for determination of the expression of C/EBP homologous protein (CHOP),Bcl-2,and activated caspase-3 in the left hippocampi by Western blot.Results Compared with group S,NDSs and brain water content were significantly increased,the expression of CHOP and activated caspase-3 was up-regulated,and the expression of Bcl-2 was down-regulated in group FCIR,NDSs was increased in group P (P < 0.05).Compared with group FCIR,NDSs and brain water content were significantly decreased,the expression of CHOP and activated caspase-3 was down-regulated,and the expression of Bcl-2 was up-regulated in group P,and no significant change was found in each parameter in group Ⅰ (P > 0.05).Conclusion Propofol can reduce focal cerebral I/R injury through inhibition of the endoplasmic reticulum stress-mediated apoptosis in rats.
4.Feasibility of amplification refractory mutation system in fast detection of clarithromycin resistance of Helicobacter pylori in gastric mucosa
Jiang LI ; Chunfeng CHEN ; Weixiang SHEN ; Xiaoyan ZHANG ; Wen GAO ; Haihui SHENG ; Hengjun GAO ; Hong CHENG
Chinese Journal of Digestion 2017;37(9):593-597
Objective To evaluate the feasibility of the combination of amplification refractory mutation system (ARMS) and quantitative real-time polymerase chain reaction (PCR) method in fast detection of clarithromycin resistance of Helicobacter pylori (H.pylori) in gastric mucosa.Methods A total of 150 gastric mucosal specimens with positive H.pylori culture were collected from the H.pylori positive patients who failed in H.pylori eradication from January to August in 2013.The drug resistant gene mutation types of H.pylori in these samples were detected by quantitative real-time PCR based on ARMS.And the accuracy was confirmed by sequencing.The clarithromycin resistance of H.pylori was determined by E-assay.Chi-square test was used for statistical analysis.Results Among 149 gastric mucosal specimens (one specimens without wild type or mutation type had been eliminated),the results of quantitative real-time PCR based on ARMS of two samples were not consistent with the results of sequencing;the consistent rate was 98.7% (147/149).Among 149 specimens with positive H.pylori culture,104 samples (69.8%) were clarithromycin resistance.In 101 samples the clarithromycin resistance was detected by quantitative real-time PCR based on ARMS;the consistent rate was 97.1% (101/104).Both E-assay and clarithromycin resistant rate detected by E-assay or quantitative real-time PCR based on ARMS was 69.8% (104/149) and 67.8% (101/149),respectively,and the difference was not significant (x2 =0.141,P=0.932).Conclusion The combination of ARMS and quantitative real-time PCR method in fast detection of clarithromycin resistance of H.pylori in gastric mucosa is strongly feasible and highly consistent has high consistent rate with sequencing and E-assay.
5.Preliminary clinical application of catheter - directed thrombolysis with urokinase for acute limb ischemia
Yujin LIU ; Xiaojun ZHANG ; Weixiang SONG ; Xubo SHEN ; Liubo LI
Journal of Interventional Radiology 2018;27(3):262-265
Objective To evaluate the curative effect and feasibility of catheter-directed thrombolysis with urokinase for acute limb ischemia. Methods The clinical data of a total of 21 patients with acute limb ischemia, who were treated with infusion of urokinase through UniFuse thrombolytic catheter, were retrospectively analyzed. The clinical curative efficacy, prognosis and complications of catheter - directed thrombolysis (CDT) were summarized. Results In all patients, thrombolysis therapy was effective, the clinical symptoms were alleviated, and successful limb salvage was achieved. One patient died of cerebral hemorrhage due to renal decompensation. No other serious complications such as hemorrhage, hematoma or disability occurred. Conclusion Preliminary clinical study shows that CDT with urokinase is effective and safe for the treatment of acute limb ischemia. This therapy can help reduce unnecessary surgical procedures or stent implantation, and the risk of thrombolysis is controllable. (J Intervent Radiol, 2018, 27: 262-265)