1.Effects of tumor suppressor NDRG2 gene on proliferation and invasive ability of colon cancer cell line SW620
Jianxun WANG ; Tian ZENG ; Peng ZHANG ; Zhigang WANG ; Hailu LEI
Journal of International Oncology 2016;43(6):401-404
Objective To observe the influence of N-myc downstream-regulated gene 2 (NDRG2) on the growth and invasive ability of human colon cancer cell line SW620,and to explore its mechanism.Methods pcDNA3.1-NDRG2 and siRNA-NDRG2 were transfected transiently respectively into SW620 by Lipofectamine TM 2000,untreated cells as the control group.Western blotting was used to investigate the expression of NDRG2 and matrix metalloproteinase-2 (MMP-2).Matrigel invasion assay was used to study the invasive abilities of SW620 cells in all groups.The growth curve was determined through 3-(4,5-dimethyl-2-thiazoly)-2,5-diphenyl-2H-tetrazolium bromide method.Result After transfecting pcDNA3.1-NDRG2 into the SW620 cells,the protein level of NDRG2 increased and the expression of MMP-2 declined markedly.After transfecting siRNA-NDRG2 into the SW620 cells,the protein level of NDRG2 declined and the expression of MMP-2 increased markedly.In addition,compared with the control group (75.80 ± 4.82),the numbers of transmembrane cells in pcDNA3.1 group (56.20 ± 7.40) and in siRNA group (94.20 ± 9.23) were significantly different (t =13.102,P =0.000;t =11.820,P =0.000).The growth curve showed that:compared with the control group (0.67 ±0.01),the absorbance of the fifth day after transfection in pcDNA3.1 group (0.46 ±0.01) and in siRNA group (0.91 ± 0.02) were different significantly (t =9.561,P =0.000;t =10.922,P =0.000).Conclusion NDRG2 can reduce the invasion and proliferation ability of colon cancer cell SW620,and its mechanism may be related to the down-regulation of MMP-2 expression.
2.Protective effect of aqueous extract of velvet antler on cisplatin-induced nephrotoxicity in mice
Simin DONG ; Hailu WANG ; Quankai WANG ; Jing ZHANG
Chinese Journal of Pathophysiology 2016;32(8):1466-1470
AIM:To study the protective effect of aqueous extract of 2-branched and 3-branched velvet antler on cisplatin (CDDP)-induced nephrotoxicity in mice .METHODS:The mouse model of renal injury was induced by intra-gastric administration of CDDP at the dose of 15 mg/kg.After treatment, kidney index (KI), serum creatinine (SCr), blood urea nitrogen (BUN), the activity of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), and the content of malondialdehyde ( MDA) in the kidney were determined .The renal pathological changes were observed with HE staining.RESULTS:Aqueous extract of velvet antler at the tested dose markedly decreased BUN , SCr and the content of MDA, and elevated the activity of SOD and GSH-Px in the mice pretreated with CDDP ( P<0.05) .The pathological chan-ges of the renal tissues were improved obviously , and the injury of the epithelial cells of renal tubules was mitigated .The effect of the aqueous extract of 2-branched velvet antler on renal function and cisplatin-induced nephrotoxicity was better than that of 3-branched one at the same concentration .CONCLUSION: The aqueous extract of 2-branched and 3-branched velvet antler has a certain protective effect on cisplatin-induced nephrotoxicity , which may be associated with in-creasing the anti-oxidative capability of mouse renal tissue .
3.Sequence analysis of p62dok amino acid and cDNA clone
Changhua WANG ; Feng DONG ; Chuanren DONG ; Hanqiao ZHENG ; Hailu YANG
Chinese Journal of Pathophysiology 2001;17(5):407-410
AIM:To analyse sequences of p62dok amino acid and cDNA and to investigate p62dok tyrosine phosphorylation and its relation with p21ras GAP. METHODS:The purified p62dok was extracted from CHO/IR cells. The peptide sequence of p62dok was carried out on a high performance analyzer. PCR was performed with the primers designed from the sequence of p62dok amino acid. Western blot and immunoprecipitation were used to identify tyrosine phosphorylation of p62dok and the binding of p62dok with p21ras GAP. RESULTS:The p62dok cDNA is a 1863 bp sequence and code 481 amino acid with 15 tyrosine residues and a putative pleckstrin homology domain. The p62dok protein is rich in PxxP motif. The tyrosine-phosphorylated p62dok can bind p21ras GAP. CONCLUSION:Perhaps p62dok is a new signaling molecule and play an important role in insulin signaling networks through RAS/MAPK pathway.
4.Sequence analysis of p62~(dok) amino acid and cDNA clone
Changhua WANG ; Feng DONG ; Chuanren DONG ; Hanqiao ZHENG ; Hailu YANG
Chinese Journal of Pathophysiology 1989;0(05):-
AIM:To analyse sequences of p62 dok amino acid and cDNA and to investigate p62 dok tyrosine phosphorylation and its relation with p21 ras GAP. METHODS:The purified p62 dok was extracted from CHO/IR cells. The peptide sequence of p62 dok was carried out on a high performance analyzer. PCR was performed with the primers designed from the sequence of p62 dok amino acid. Western blot and immunoprecipitation were used to identify tyrosine phosphorylation of p62 dok and the binding of p62 dok with p21 ras GAP. RESULTS:The p62 dok cDNA is a 1863 bp sequence and code 481 amino acid with 15 tyrosine residues and a putative pleckstrin homology domain. The p62 dok protein is rich in PxxP motif. The tyrosine-phosphorylated p62 dok can bind p21 ras GAP. CONCLUSION:Perhaps p62 dok is a new signaling molecule and play an important role in insulin signaling networks through RAS/MAPK pathway.
5.Crystal form identification of dasatinib in tablets
Jianming WANG ; Yanmei CHEN ; Hailu ZHANG ; Yanfeng ZHANG ; Minhua CHEN
Journal of China Pharmaceutical University 2015;(5):575-578
The aim of this study was to establish the methods to identify crystal form of dasatinib in tablets.X-ray powder diffraction(XRPD)and solid-state nuclear magnetic resonance(ssNMR)were used to analyze the crystal form of dasatinib in Sprycel? tablets and Yinishu? tablets.The results showed that monohydrate and anhydrate were identified in Sprycel? and Yinishu? tablets respectively;with no detectable anhydrate in Sprycel? tablets and no detectable monohydrate in Yinishu? tablets.The results of XRPD and ssNMR were consistent;and could be both applied in the crystal form identification of dasatinib in tablets.
6.Effects of thyroid hormone on the myosin heavy chain mRNA expression in cardiac myocytes induced by angiotensin Ⅱ
Baohua WANG ; Jingping OUYANG ; Yongming LIU ; Hanqiao ZHENG ; Lei WEI ; Jingwei YANG ; Ke LI ; Hailu YANG
Chinese Journal of Pathophysiology 2000;0(11):-
AIM: To study the effect of thyroid hormone on the expressional change of myosin heavy chain(MHC) gene in cardiomyocyte induced by angiotensinⅡ(AngⅡ) and its potential mechanism. METHODS: Cardiac myocyte was cultured according to the method of Simpson. 10 -8 mol/L T_3 and 10 -7 mol/L AngⅡ were added to the culture medium,respectively or synchronously. After 48 h,the expression of ? and ?-MHC mRNA in myocytes were detected by RT-PCR. The protein kinase C activation were detected by PepTag non-radioactive PKC assay. The incorporation of -Leucine and -thymine to test the protein and DNA synthesis in myocytes were also performed. RESULTS: AngⅡalone increased the incorporation of -Leucine of myocytes while it had no effect on the incorporation of -thy mine. The expression of ?-MHC mRNA was increased and the expression of ?-MHC mRNA was decreased significantly at the condition of AngⅡ. The enhanced PKC activation was induced by AngⅡalso. When AngⅡand T_3 were added to the culture medium synchronously,though the incorporation of -leucine and -thymine were not changed compared with AngⅡ treated alone. The ?-MHC mRNA expression was increased and the ?-MHC mRNA expression was decreased significantly. The PKC activation of the myocytes also was decreased. CONCLUSIONS: T_3 inhibited the expressional change of myosin heavy chain gene in cardiac myocytes induced by AngⅡ. The effect of T_3 on the change of PKC activation in cardiac myocytes may be one of its mechanisms.
7.Protective effects of multi-enzyme Ⅱ on vascular endothelial cell injury induced by hyperlipidemia serum
Yongming LIU ; Jingping OUYANG ; Shuzhen TU ; Hanqiao ZHENG ; Jingwei YANG ; Hailu YANG ; Hailong WANG
Chinese Journal of Pathophysiology 2000;0(11):-
AIM and METHODS: The protective effects of multi-enzyme Ⅱ was studied on cultured endothelial cells which was injuried by hyperlipidemia serum. RESULTS: Hyperlipidemia serum increased ICAM-1 expression on the surface of endothelial cells, and decreased NO- 2 release significantly (P
8.Protective effects and its mechanism of rebamipide on aspirin-induced injury in human gastric mucosal epithelium cells
Zhaotao DUAN ; Zhenyu ZHANG ; Hailu WU ; Fangcen YUAN ; Zongdan JIANG ; Bangshun HE ; Shukui WANG
Chinese Journal of Digestion 2014;34(7):453-457
Objective To investigate the protective effects and its mechanism of rebamipide on aspirin-induced injury in human gastric mucosal epithelium cells (GES-1).Methods GES-1 cells monolayer culture model was established in vitro.Then the cells were divided into negative control group,aspirin injured group and combination of rebamipide at different concentration (0.2,0.5,1.0 mrnol/L) and aspirin groups.The cell proliferation,the content of malondialdehyde (MDA) and the activity of superoxide dismutase (SOD) of each group were detected.The ultrastructural changes of each group were observed by transmission electron microscopy (TEM).The expressions of nuclear factor erythroid 2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) at protein level in the cells of each group were detected by Western blot.Nrf2 interfering suppression test was performed and then the influence of Nrf2 small interfering RNA (siRNA) on the expression of HO-1 protein was observed.One-way analysis of variance was performed for comparison among multi-groups and t-test was used for comparison between the two groups.Results The cell viability of aspirin injured group and combination of rebamipide at different concentration (0.2,0.5,1.0 mmol/L) and aspirin groups were (49.56±3.88)%,(59.34±4.36) %,(70.79 ± 5.96) % and (86.07 ± 5.20) %,respectively,and the difference was statistically significant (F=30.634,P< 0.01).Compared with aspirin injured group,the content of MDA significantly lowered in combination of rebamipide at different concentration (0.2,0.5,1.0 mmol/L) and aspirin groups ((2.26±0.25) nrnol/rng vs (1.85±0.13) nmol/mg vs (1.62±0.11) nmol/mg vs (1.13±0.15) nmol/mg),and the difference was statistically significant (F=23.821,P<0.05).Compared with aspirin injured group,the activity of SOD significantly increased in combination of rebamipide at 0.5 and 1.0 mmol/L and aspirin groups ((8.49±0.89) U/rng vs (11.50±1.03) U/mg vs (13.74±0.76) U/mg),the difference was statistically significant (F=25.666,P<0.05).Under TEM,the cell ultrastrucmral was obviously inured in aspirin treated,while rebamipide could relieve the injury.The differences of relative expression quantity of Nrf2 and HO-1 at protein level among combination of rebamipide at 0.2,0.5 and 1.0 mmol/L and aspirin groups and aspirin injured group were statistically significant (0.35±0.04 vs 0.46± 0.05 vs 0.84±0.08 vs 0.15±0.02,0.72±0.09 vs 0.93±0.11 vs 1.29±0.14 vs 0.39±0.07,F=92.550and 38.235,both P<0.05).After transfected with Nrf2 siRNA,the expression of HO-1 was 0.38±0.04 in aspirin injured group and 0.62±0.08 in combination of rebamipide and aspirin group,which was lower than that before transfection (0.61 ± 0.05,1.33± 0.09),respectively.The differences were statistically significant (t =6.276 and 10.444,both P<0.05).Conclusion Rebamipide may activate Nrf2/HO-1 pathway and relieve aspiriwinduced oxidative stress in GF1 ceils.
9.Study of laparoscopic procedure for remove of common bile duct stone and T-tube drainage
Hailu LEI ; Ziqiang YAN ; Kangtai ZHANG ; Kecheng WANG ; Mingfu DENG ; Lixun PENG ; Shaodi FAN
Chinese Journal of General Surgery 1993;0(02):-
Objective To study the optimal laparoscopic procedure and its indication for remove of common bile duct stone. Methods Analysis was made on the clmical data of 124 cases of laparoscopic choledocholithotomy and T tube drainage in our center.Results 82 patients underwent the improved laparoscopic procedure, alternation to open operation in 4 cases (4.9%),and the mean operating time was (80?30) min. While 42 patients were operated with traditional laparoscopic method,changing to open operation in 6 cases (14.3%),and the mean operating time was (170?40) min . The improved method could shorten the operation time and reduce the open operation rate significantly than traditional method did (P
10.Glaucocalyxin A: A Review
Yuqian LU ; Qingchao TU ; Hailu YAO ; Jigang WANG ; Fei XIA
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(22):249-258
Glaucocalyxin A (GLA) is a natural diterpenoid extracted from Isodon amethystoides belonging to Labiatae. Modern pharmacological research has shown that GLA has anti-inflammatory, anti-tumor, anti-fibrotic, osteoporosis-ameliorating, and cardiovascular system-protecting activities and good biosafety. However, the low content in plants, poor solubility, high metabolic rate, and low bioavailability limit the application of GLA. To address these issues, researchers have studied the total synthesis, structural modification, and nanomedicine development of GLA. By reviewing the available studies about GLA in the past five years, we summarize the research progress in the total synthesis, pharmacological activities and mechanisms, and in vivo metabolic transformation, aiming to provide a theoretical basis for clarifying the specific mechanisms underlying the pharmacological activities of GLA and for further research, development, and clinical applications of GLA.