1.IKKαregulates ultraviolet radiation-induced activation of p53 in a p38K-dependent manner
Hongli WANG ; Yongliang HU ; Meiru HU ; Lun SONG ; Yuanfang MA
Military Medical Sciences 2015;(5):321-324
Objective To explore the signal transduction mechanism of inhibitor kappa B kinase α( IKKα) , one of the catalytic subunits of IKK complex , for regulating p53 transactivation in the cellular ultraviolet radiation ( UVB) repsonse. Methods The transactivation of p53 was determined by dual-luciferase reporter gene analysis system while the expression and activation of IKKα, IKKβ, p53 and p38K was detected by Western blotting assay .Results UVB exposure induced activation and transactivation of p 53 in the wild type mouse fibroblasts ,but the effect was blocked by IKKa deficiency and recovered by reconstitution of IKKαexpression.Under the same conditions , IKKαregulated p38K activation, while inhibi-ting p38K activation down-regulated p53 transactivation under UVB exposure .Conclusion IKKαregulates UVB-induced phosphorylation and activation of p 53 in a p38K-dependent manner .
2.Cloning and expression of ns5a gene of hepatitis C virus 1b strain DY in Escherichia coli
Hui FU ; Yonglie CHU ; Lili ZHANG ; Meiru CAO ; Fan CHENG ; Xiaorong RU ; Yongjian WANG
Journal of Xi'an Jiaotong University(Medical Sciences) 1982;0(04):-
Objective To clone and express the ns5a gene of hepatitis C virus(HCV) 1b strain DY.Methods By using the prokaryotic cell gene engineering,HCV ns5a gene was amplified with nested PCR from the plasmid HCV17 of HCV 1b strain DY full-length gene and inserted into the cloning pMD18-T vector.The cloned HCV ns5a gene was separated and subcloned into expression vector pET-28a and induced by IPTG in E.coli.BL21.The expressed product was identified by SDS-PAGE and Western-blot methods.Results Recombinant expression plasmid pet-28a-ns5a was constructed and expressed successfully.Conclusion HCV ns5a gene was cloned and expressed.This might be helpful for further studies on the nature and biological properties of the ns5a gene.
3.PM2.5 induces VEGF expression and inflammatory responses by trans-activating AP-1 in human bronchial epithelial cells
Xiuduan XU ; Shasha LIU ; Hongli WANG ; Yongliang HU ; Meiru HU ; Lun SONG
Military Medical Sciences 2016;40(7):541-544
Objective To explore the role of the transcriptional factor activator protein (AP)-1 in mediating vascular endothelial growth factor ( VEGF) expression in human bronchial epithelial cells exposed to PM 2.5.Methods Beas-2B cells was treated with PM2.5.Luciferase assay was used to detect the activation status of AP-1 and transcription of VEGF in the Beas-2B cells.The induced activation of c-Jun, ATF2 and VEGF expression was tested by Western blotting assay.Results PM2.5 induced transactivation of the transcriptional factor AP-1, accompanied by phosphorylation of the AP-1 components, c-Jun and ATF2 in Beas-2B cells.Moreover, when AP-1 activation was inhibited by knocking down c-Jun or ATF2 expressions, induction of VEGF expression was partially attenuated in Beas-2B cells.Conclusion AP-1 is a critical transcriptional factor in mediating PM2.5-induced VEGF expression and inflammatory responses in human bronchial epithelial cells.
4.Effect of kojic acid on immune systems of mice damaged by gamma-irradiation
Kai WANG ; Pengfei LI ; Chunguang HAN ; Li DU ; Meiru YUAN ; Chao LIU ; Yongxue LIU
Military Medical Sciences 2014;(10):771-774
Objective To evaluate the effect of kojic acid( KA) on the immune system of mice after exposure to gam-ma-irradiation.Methods Twenty male C57BL/6 mice were divided into normal group, irradiation group, low/high doses of KA pretreated groups.Mice in normal group did not receive any treatment,while mice in other groups were sc injected with a single dose of sterile distilled water or KA(75 and 300 mg/kg, respectively) 27 h prior to a sublethal dose(4 Gy, 138.54 and 140.30 cGy/min, respectively) of whole body gamma-irradiation.The injected volume was calculated by 0.2 ml/20 g.Forty mice were sacrificed at day 2 and day 8 post-irradiation, respectively.The splenic lymphocyte transformation and spleen and thymus indexes were determined.Histopathological sections were produced, and the morphological changes were also observed.Results The splenic lymphocyte transformation capacity and spleen and thymus indexes of mice pre-treated with 300 mg/kg elatve mass KA were increased significantly ( P<0.01) compared with the irradiation group.The morphological changes in the spleen and thymus of mice in 300 mg/kg KA pretreated group were better than in the irradia-tion group.The above parameters of mice in irradiation group were injured severely in comparison with the normal group. Conclusion Acute radiation can damage the immune system of mice obviously.KA can enhance the transformation capaci-ty of lymphocytes and has marked protective effect on the immune system of mice after irradiation.
5.The control study of treatment between dual-antiplatelet aggregation and warfarin in the prevention of thromboembolism in high risk patients of nonvalvular atrial fibrillation GENG
Qingfeng GE ; Meiru LI ; Hongyue LI ; Bin WANG ; Shuzhang AN ; Lijuan CHENG ; Jianti QI ; Yuhui LIU ; Yongli YANG ; Shun CHANG
Chinese Journal of Postgraduates of Medicine 2010;33(1):10-12
Objective To compare the efficiency and safety of aspirin-dipyridamole and warfarin in the prevention of thromboembolism in patients with nonvalvular atrial fibrillation(NVAF)and high risk factors.Methods One hundred and forty NVAF patients with high risk factors were randomly divided into two groups.Warfarin group[78 cases international normalized ratio(INR)2.0-3.0,for the patients older than 75 years,INR ranging from 1.6 to 2.5]and combination group(62 cases received aspirin 160 mg once every day plus dipyridamole 160 mg 3 times every day).The incidence of death,thromboembolism(including stroke and peripheral arteries embolism)and hemorrhage events were observed.Results Followed-up 12-28 months.In warfarin group,3 cases lost,2 cages had stroke,2 cases suffered from serious bleeding events,6 cases had minor bleeding events.In combination group,2 cases lost,6 cases had stroke,and 2 cases suffered from peripheral arteries embolism events,3 cases had minor bleeding events,but no serious bleeding events occurred.The incidence of thromboembolism in warfarin group wag,lower than that in combination group[2.7%(2/75)vs 13.3%(8/60),P<0.05].There was no significant difference of the bleeding rate between the two groups[10.7%(8/75)vs 5.0%(3/60),P>0.05].Conclusions Warfarin anticoagulative therapy is more effective than aspirin and dipyridamole antiplatelet dual therapy for the prevention of thromboembolism events in patients with NVAF and high risk factors.The major bleeding events in warfarin group occurs in patients with INR>3.0,so under intensive monitoring(INR 2.0-3.0),warfarin therapy is effective and safety.
6.Improvement in anesthesia method for lumbar spine surgery: esketamine-based anesthesia
Meiru WANG ; Bin SUN ; Dandan SU ; Qianwen ZHAO ; Yao LU ; Huanjia XUE ; Liwei WANG
Chinese Journal of Anesthesiology 2023;43(9):1082-1086
Objective:To evaluate the efficacy of esketamine-based anesthesia in lumbar spine surgery.Methods:Ninety-four patients of both sexes, aged 18-64 yr, with body mass index of 18.5-29.9 kg/m 2, of American Society of Anesthesiologists Physical Status classification ⅠorⅡ, scheduled for elective lumbar posterior decompression bone grafting fusion internal fixation under general anesthesia from June 2022 to December 2022, were divided into control group(group C) and esketamine group(group K) using a random number table method, with 47 cases in each group. Midazolamm, sufentanil, etomidate and cisatracurium were intravenously injected for anesthesia induction in both groups, and esketamine 0.5 mg/kg was intravenously injected on this basis in group K. Propofol and remifentanil were intravenously infused to maintain anesthesia, and cisatracurium besylate was intermittently injected to maintain muscle relaxation in both groups, and esketamine 0.25 mg·kg -1·h -1 was intravenously infused on this basis in group K. The patients were connected to an analgesic pump for patient-controlled intravenous analgesia at 10 min before the end of surgery, and flurbiprofen axetil 50 mg was intravenously injected for rescue analgesia when the numeric rating scale score >4. The time of first pressing the analgesia pump, effective pressing times of the analgesia pump within 48 h after operation and requirement for rescue analgesia were recorded. The initial dose of remifentanil, cumulative amount of remifentanil used during operation, time of tracheal extubation, and adverse reactions within 48 h after surgery were recorded. Results:Compared with group C, the cumulative use of remifentanil during operation was significantly reduced, the time of first pressing the self-control button of the analgesia pump after surgery was prolonged, the pressing times of the analgesia pumps were decreased( P<0.05), and no significant change was found in terms of the initial dose of intraoperative remifentanil, rate of postoperative rescue analgesia, time of extubation, and incidence of adverse reactions after surgery in group K( P>0.05). Conclusions:Esketamine-based anesthesia can reduce the amount of intraoperative opioids, delay the time of postoperative pain and reduce the early postoperative pain when used for lumbar spine surgery.
7.Comparative Study of Cytotoxicity and Anti-inflammatory Effects between Raw Aconitium kusnezoffii and Aconitium kusnezoffii Processed with Terminalia chebula
Meiru ZHI ; Shu HAN ; Kaiyang LIU ; Xitao HAN ; Yanan TANG ; Ziqin LIU ; Hongyue WANG ; Fei LI ; Hong DU
China Pharmacy 2020;31(22):2701-2705
OBJECTIVE:To comp are cytotoxicity and anti-inflammatory effects of raw Aconitium kusnezoffii and A. kusnezoffii processed with Terminalia chebula . METHODS :Using H 9c2 cardiomyocytes isolated from rat as subjects ,CCK-8 assay was used to detect the effects of 0.5,1,2,4,6,8,10 mg/mL raw A. kusnezoffii and A. kusnezoffii processed with T. chebula on cell inhibition rate after cultured for 4,8,12,24 h. Hoechst 33258 staining was used to observe the effects on cell morphology characteristics after treated with 2,4,6 mg/mL raw A. kusnezoffii and A. kusnezoffii processed with T. chebula . Using macrophages RAW264.7 cells as subjects ,CCK-8 assay was used to detect the effects of 0.05,0.1,0.25,0.5,0.75,1,1.5,2 mg/mL raw A. kusnezoffii and A. kusnezoffii processed with T. chebula on cell survival rate after cultured for 24 h. ELISA assay was used to detect the effects of 0.05,0.1,0.25,0.5 mg/mL raw A. kusnezoffii and A. kusnezoffii processed with T. chebula on the release of NO , TNF-α and IL-6 in RAW 264.7 inflammation cells induced by LPS. RESULTS :When the mass concentration was 0.5,1 mg/mL, neither raw A. kusnezoffii and A. kusnezoffii processed with T. chebula had no inhibitory effect on H 9c2 cells. When the mass concentration was 2 mg/mL,the inhibitory effects of A. kusnezoffii processed with T. chebula on H 9c2 cells was higher than that of raw A. kusnezoffii (P<0.05 or P<0.01);fluorescence intensity of cells treated for 24 h was stronger than that of raw A. kusnezoffii,but its nucleus was intact. When the mass concentration was 4-10 mg/mL,the inhibitory rate of A. kusnezoffii processed with T. chebula on H 9c2 cells at different time points (except for 24 h culture of 8,10 mg/mL)was significantly lower than raw A. kusnezoffii (P<0.05 or P<0.01). The characteristics of cell morphology also showed that the fluorescence intensity of raw A. kusnezoffii group at 4,6 mg/mL was stronger than that of A. kusnezoffii processed with T. chebula group,and the cell nucleus fragmentation was more serious in the raw A. kusnezoffii group. 0.05-0.5 mg/mL raw A. kusnezoffii and A. kusnezoffii processed with T. chebula had no toxicity to RAW264.7 cells. 0.25,0.5 mg/mL raw A. kusnezoffii and 0.1,0.25,0.5 mg/mL A. kusnezoffii processed with T. chebula showed significant inhibitory effect on the release of NO ,0.05,0.1,0.25,0.5 mg/mL raw A. kusnezoffii and A. kusnezoffii processed with T. chebula showed significant inhibitory effect on the release of TNF-α and IL-6 in RAW 264.7 cell(P<0.05 or P< 0.01). The inhibitory effects of A. kusnezoffii processed with T. chebula at the same concentration on the release of NO was better than that of raw A. kusnezoffii ,but poorer than raw A. kusnezoffii in the inhibitory effects on the release of TNF-α and IL-6. CONCLUSIONS:The toxicity of A. kusnezoffii can be reduced after processed with T. chebula ,especially the toxicity of it in medium or high concentration and short-term use is lower than that of raw A. kusnezoffii . At the same time ,the anti-inflammatory effect of A. kusnezoffii processed with T. chebula is comparable to that of raw A. kusnezoffii at the same concentration.