1.Effects of N-acetylcysteine on the expression of cardiac adiponectin and its receptors in diabetic rats
Zhixin GUO ; Caihong ZHENG ; Ping WEI
Medical Journal of Chinese People's Liberation Army 1981;0(04):-
Objective To investigate the effects of N-acetylcysteine (NAC) on the expression of cardiac adiponectin and its receptors in streptozotocin-induced diabetic rats. Methods Thirty-two male Wistar rats were randomly divided into four groups, with 8 in each: untreated control group (C group), NAC-treatment group (CT group), diabetic untreated group (D group) and diabetic NAC-treatment group (DT group). After 8 weeks, plasma glucose and insulin, and cardiomyocyte cross sectional area were measured. Cardiac protein expression of adiponectin receptor 1 and 2 (adipoR1 and adipoR2), AMP-activated protein kinase ? (AMPK?), phosphorylation of AMP-activated protein kinase ?-subunits and glucose transporter 4 (GluT4) were determined by Western blotting. Plasma and myocardial adiponectin levels were measured by radioimmunoassay. Plasma and myocardial free 15-F2t-isoprostane levels were assayed by enzyme immunoassay. Results Compared with C group, the ratio of ventricular weight to body weight and cardiomyocyte cross-sectional area, plasma and levels of free 15-F2t-isoprostane in myocardium and the protein expression of myocardial adipoR1 increased significantly in diabetic rats (D group) (P
2.Applied research of deep hypothermic circulatory arrest and regional cerebral perfusion in pediatric aortic arch surgery
Zheng GUO ; Jinghao ZHENG ; Wei WANG ; Weiding FU ; Deming ZHU
Chinese Journal of Thoracic and Cardiovascular Surgery 2014;30(6):363-365
Objective To observe and evaluate the effects of the deep hypothermic circulatory arrest(DHCA) and regional cerebral perfusion(RCP) in pediatric aortic arch surgery.Methods According to different methods of CPB,70 infants less than 3-month-old with CoA or IAA were undergone corrective surgery with DHCA or RCP.The bypass time,aortic clamp time,DHCA or RCP time,ventilation time,ICU stay time and post-operative complications were recorded and compared between two groups.Results The incidence of neurological complications was significantly higher in DHCA group.The CPB time was significantly longer in the RCP group,and the RCP time was significantly longer than DHCA time.Blocking time,ventilator intubation time,ICU residence time,postoperative renal dysfunction,low cardiac output,puhnonary inflammation and hospital mortality was no significant difference between the two groups.Conclusion RCP is an effective cerebral protection technique.Compared with DHCA,RCP works better in sustained brain cerebral perfusion and is suitable for complex aortic arch operation in children.It has a better effort in protection of the neurological system than DHCA.
3.Study of Isolating Anti-Endotoxin Monomer Component from Radix Paeoniae Rubra Biosensor by Biosensor Technique
Genfa LU ; Guo WEI ; Yibin GUO ; Jiang ZHENG ; Guangxia XIAO
China Pharmacy 1991;0(04):-
OBJECTIVE:To isolate anti-endotoxin monomer component from Radix Paeoniae Rubra by means of biosen?sor technique.METHODS:The surface of biosensor cuvette was embedded by Lipid A;the screening target was established,tracking the silica gel column chromatogram and the binding ability of effluent component from HPLC with Lipid A with the ultraviolet scan result of the reclaimed material from biosensor as reference;anti-endotoxin monomer component was isolated;the component of monomer and the synthetic action of extrinsic lipopolysaccharides were also assayed by LAL test method.RESULTS:Components binding to Lipid A was reclaimed from cuvetee by biosensor technique,with the wavelength of UV absorption peak at194nm,215nm and275nm respectively.Anti-endotoxin monomers of higher binding activity with Lipid A isolated by HPLC method were1,2,3,4,6—O—pentagalloyl—?—D—glucose(PGG).PGG at concentration of8,4,2?g/ml respectively neutralized68.8%,43.7%and31.4%of LPS at an activity of0.1EU/ml respectively.CONCLUSION:It is fea?sible to isolate anti-endotoxin monomer component from Radix Paeoniae Rubra by means of biosensor technique,which is a fast,accurate and efficient and can be used to isolate anti-endotoxin monomer component from Radix Paeoniae Rubra on a large scale.
4.Expression and biological function identification of TCR?9/?2-Fc protein in baculovirus vector expression system
Yang GUO ; Jing ZHENG ; Yu HU ; Lianxian CUI ; Wei HE
Basic & Clinical Medicine 2006;0(12):-
Objective To establish an expression system of TCR?9/?2-Fc protein by baculovirus vector expression system and identify biological function of expressed TCR?9/?2-Fc protein.Methods ?9Fc and ?2(OT3)Fc gene fragments were amplified by overlap PCR and inserted into expression vector pBACp10ph.The recombinant plasmid pBACp10ph-?9/?2(OT3)-Fc and the baculovirus DNA were co-transfected into sf9 cells.The expression position of TCR?9/?2(OT3)-Fc was identified by Western blot and the expression efficiency of ?9Fc and ?2(OT3)Fc was tested by flow cytometry(FCM).Furthermore,the binding activity of TCR?9/?2(OT3)-Fc protein with SKOV3 cells and MNS protein was evaluated with laser scanning confocal microslopy and surface plasmon resonance(SPR).Results The recombinant vector pBACp10ph-?9/?2(OT3)-Fc was constructed and TCR?9/?2(OT3)-Fc protein was expressed in sf9 cells.However,the expression efficiency of ?9Fc and ?2(OT3)Fc was quite different.It was proved that purified TCR?9/?2(OT3)-Fc protein can bind with SKOV3 cell and MNS protein.Conclusion TCR?9/?2-Fc protein is successfully expressed in baculovirus vector expression system and TCR?9/?2-Fc protein can simulate the binding activity of TCR in vitro.
5.Gene expression profiles analysis identifies key genes of PBMCs in patients with benign and malignant breast tumor
Lang HE ; Na WEI ; Zheng GUO ; Dan WANG
Chinese Journal of Immunology 2016;32(10):1424-1427,1436
Objective:To observe the changes of gene expression in peripheral blood mononuclear cells( PBMCs) of benign and malignant breast tumor based on gene expression profiling. Methods: Datasets of gene expression profiling were downloaded from the GEO database,including PBMCs profilings of benign breast tumor,breast cancer and healthy controls. GEO2R tool was used to analyze the data to identify the differentially expressed genes (DEGs). Function of DEGs were annotated by DAVID. Protein interaction analysis and hub gene select were then performed using STRING database. Results:563 and 237 DEGs respectively were identified. DEGs in breast cancer involved in biological process of leukocyte activation,angiogenesis and leukocyte transendothelial migration. The hub genes are IL8,RHOB,ITGB1. Conclusion:The data suggests that gene expression patterns of these two profilings are different at a certain degree. PBMCs maybe a better noninvasive material for biomarker detection of benign and malignant breast tumor.
6.Effects of matrine on expression of a proliferation-inducing ligand in colorectal cancer cell lines
Zhuofu WEN ; Yunwei GUO ; Yongwei LI ; Fengping ZHENG ; Xiuqing WEI
Chinese Journal of Digestion 2008;28(9):621-624
Objective To study the effect of matrine on the expression of a proliferation-inducing ligand (APRIL) in colorectal cancer cell line (SW480 cell). Methods MTT assay was used to evaluate the inhibitory effect of matrine on SW480 cells. The protein and mRNA levels of APRIL in SW480 cells were determined by immunohistochemistry and real-time fluorescence quantitative PCR (RFQ-PCR). SW480 cells were treated with 0.5,1.0,2.0 mg/ml of matrine for 24 h, 48 h and 72 h. FU and blank were served as drug control and blank control groups, respectively. Results Matrine had obviously inhibitory effect on proliferation of SW480 cells in a time- and dose-dependant manner. The expression of APRIL was strong in SW480 cells. When treated with 50,100,200 ug/ml of FU, the APRIL mRNA levels in SW480 cells raised gradually and reached the highest levels at 72 h after treatment, which were significantly higher than those in blank control group (all P value<0.001). When treated with 0. 5,1.0, 2.0 mg/ml of matrine, the APRIL mRNA levels in SW480 cells increased at 24 h after treatment, which were significantly higher than those in blank control group (all P value<0. 001), and then decreased gradually and almost equal to level of blank control group at 72 h. Conclusion In treatment with FU, the survival cells.may have stronger ability of proliferation due to higher expression of APRIL in SW480 cells. Anti-APRIL therapy might be an important assistant treatment to counter the impact of APRIL. Matrine will not cause persistent increase of APRIL mRNA levels in SW480 cells, so it might be a helpful drug in anti-tumor theraphy.
7.Relationship between Toll-like receptor 4(TLR4) on the peripheral blood monocytes and serum TNF-α in patients with chronic severe hepatitis B
Xiuqing WEI ; Zhuofu WEN ; Yunwei GUO ; Fengping ZHENG
Journal of Chinese Physician 2008;10(7):886-889
Objective To study the change of TLR4 on peripheral blood monoeytes (PBMCs) and its role in the pathogenesis of chronic severe hepatitis B. Methods The expression of TLR4 on 10000 CDI4 + PBMCs was determined by flow eytometer in 30 healthy control,31 patients with chronic hepatitis B and 30 patients with chronic severe hepatitis B. The level of serum tumor necrosis factor α(TNF- α) was determined by ELISA. Results The values of TLR4 on PBMCs and serum TNF-αof the groups of healthy control, patients with chronic hepatitis B and patients with chronic severe hepatitis B were 2.3±1.1,3.7±2.3, (6.9±4.1 ) mean fluorescence intensity (MFI) and (53.8±38.1 ), ( 164.3±89.9) and (359.8±140.0) ng/L. The TLR4 value in patients with chronic severe hepatitis B was signifi- cant higher than those in healthy control and the patients with chronic hepatitis B ( P <0.05). However, there was no significant difference between the patients with chronic hepatitis B and healthy control ( P > O. 05 ). TNF-α increased gradually and significantly from the healthy control to the patients with chronic hepatitis B and patients with chronic severe hepatitis B. There was a significant positive correlation be- tween the value of TLR4 and the value of serum TNF-αin the patients with chronic severe hepatitis B( r=0.666, P <0.01). Conclusion There may be a role of TLR4 in the pathogenesis of chronic severe hepatitis B.
8.Prevention of extracorporeal shock wave lithotripsy-induced renal Injury by pre -treating kidneys with low-energy shock waves
Xiqing GUI ; Zhenyu GUO ; Huabin SUN ; Wei ZHENG ; Fang YIN
Journal of Chinese Physician 2008;10(6):770-772
Objective To investigate the prevention and mechanism of Extracorporeal shock wave lithotripsy(ESW) induced renal Injury by pre-treating kidneys with low-energy shock waves(LESW).Methods Forty healthy female domestic rabbits were surgically managed to the mono-nephron models and random divided into 4 groups consisting of ten each: Control,LESW,ESW and ESWL plus LESW pretreated groups.LESW group received 100 LESW,ESW group received 1500 standard ESW,and same dose on ESW group except 100 LESW pretreatment in ESW plus LESW pretreated group.The rabbit kidney tissues were obtained 24 hours after ESW.Activity of superoxide dismutase(SOD) and malondialdehyde(MDA) levels in the renal tissue,and the level of N-acetyl-β-D-glucosaminidase(NAG) in urinary were measured.Renal cell apoptosis was detected by TdT-mediated dUTP Nick End Labelling(TUNEL).Results The MDA,the urinary level of NAG and rate of apoptosis in the LESW groups were reduced(P<0.01),and the activity of SOD increased significantly(P<0.05) as compared with ESW group,and these changes in LESW group had no statistics difference compared with the control group(P>0.05).Conclusions LESW pretreatment protocol substantially limits the renal injury that often caused by ESW.LESW may suppress oxidative stress and antagonize the process of renal cellular apoptosis.
9.Elevated Toll-like receptor 4 (TLR4) in hepatocytes in patients with chronic hepatitis B and its clinical significance
Xiuqing WEI ; Yunwei GUO ; Zhuofu WEN ; Fengpin ZHENG
Journal of Chinese Physician 2008;10(12):1641-1643
Objective To investigate the role of TLR4 in the pathogenesis of chronic hepatitis B(CHB) by study the expression of TLR4 in liver tissues in patients with CHB, and the relationship among TLR4 and serum HBV DNA level, clinical severity degrees and histo-logical grades and stages. Methods Expression of TLR4 in liver tissues was semi-quantitatively determined by immunohistochemistry and e-valuated by a scoring system in 75 patients with CHB and 10 health controls. Results The positive staining of TLR4 mainly located in the cytoplasm and some on cell membrane of bepatocytes. Expression of TLR4 in the liver tissues of patients with CHB was stronger than that of health controls. The scores of TLR4 expression in patients with mild, moderate and severe CHB were 1.0±0.5,2.3±0.5 and 2.9±0.4. The scores increased gradually and significantly along with the increase of clinical severity degrees( F = 104.8, P<0.01). The scores of TLR4 expression in the liver tissues of patients with CHB were positively correlated with the clinical severity degrees (r=0.838, P<0.01) and histological grades (r=0.579, P<0.05), but not correlated with Lg (serum HBV DNA) or histological stages. Conclusion TLR4 was up-regulated in the hepatocytes of patients with CHB. There may be a role of TLR4 in the pathogenesis of CHB.
10.Effect of peroxisome proliferator-activated receptor γ agonist on prostate epithelial cells
Lanbin ZHENG ; Yayuan ZHAO ; Wei YU ; Hui GUO ; Jie JIN
Chinese Journal of Urology 2010;31(1):52-55
Objective To assess the effect of peroxisome proliferator-activated receptor γ (PPARγ) agonist on prostate epithelial cells in vitro.Methods The expression of peroxisome proliferator-activated receptor γ(PPARγ) was studied by immunocytochemistry and immunofluorescence study.The RWPE-1 human prostate epithelial cell line was treated with PPARγ agonist rosiglitazone 100 μmol/L for 48 h.Analysis of apoptosis was performed by Caspase 3/7 activity assay.Mitochondria depolarization was measured by using the potential-sensitive color,JC-1.The expression of apoptosis-related proteins-Bax was investigated by immunohistochemistry.Results PPARγ mainly located in nucleus and perinucleus.RWPE-1 cell line treated with PPARγ agonist rosiglitazone showed higher Caspase 3/7 activity (10636±1032 RLU) than in control (5936±620 RLU),P<0.01 and significantly upregulated Bax level (8250±694 vs.6017±563)than in control group,P<0.01.In addition,mitochondrial membrane potential was depolarized in rosiglitazone treated cells.Conclusions PPARmay play important roles in the pathophysiology of BPH.The mechanism might be that PPARγ regulates cell apoptosis.It is suggested that the mitochondrial and Bax pathway might be involved in signaling PPARγ induced cell apoptosis.