1.Study on insulin resistance aggravating hypoxia/reoxygenation injury in PC12 cells by enhancing oxidative stress
Jingli REN ; Xuguang YANG ; Lei YUAN
Chinese Journal of Diabetes 2017;25(1):71-75
Objective To investigate the effects of insulin resistance (IR)on hypoxia/reoxygenation (H/R)inj ury in PC1 2 cells and the protective effects of N-actyl-L-cystine (NAC)against H/R inj ury. Methods PC12 cells were treated with 100 nmol/L insulin to induce IR. The H/R injury model was established by Na2 S2 O4. CCK-8 assay was used to detect the cell viability. Glucose consumption was detected by glucose oxidase method. Activity of superoxide dismutase (SOD)was detected by xanthine oxidase method. The levels of malonaldehyde (MDA)were measured by thiobarbituric acid method. Flow cytometry was used to determine the apoptosis and mitochondrial membrane potential (MMP). Results High-insulin inhibited insulin-induced glucose uptake in PC12 cells without affecting the cell viability (P<0. 05). PC12 cells with IR exhibited lower SOD activity and higher levels of MDA (P<0. 05 ),and enhanced apoptosis and depolarization of MMP induced by H/R(P<0. 05). NAC neutralized these effects induced by IR(P<0. 05). Conclusion IR aggravates H/R injury in PC12 cells by enhancing oxidative stress and NAC reduces the H/R injury in PC12 cells with IR via inhibiting oxidative stress and stabilizing MMP.
2.Value of serum microRNA-199a/b-3p on early diagnosis of hepatocellular carci-noma
Xuguang MI ; Lei LIU ; Shouqing LI ; Yanqiu FANG
Chinese Journal of Immunology 2015;(5):683-685,689
Objective:To analyze the expression levels of serum microRNA-199a/b-3p ( miR-199-3p) in hepatocellular carcinoma (HCC) patients and to explore whether miR-199a/b-3p could be a novel biomarker of early HCC.Methods: Patients with normal liver,liver cirrhosis and early HCC in clinic were included in this study.Blood samples of the patients were collected for analy-sis.Real time quantitative PCR technique was used to measure the expression level of serum miRNA-199a/b-3p.The AFP expression was got from clinical data.Results: Comparing with normal liver and liver cirrhosis patients, miRNA-199a/b-3p presented lower expressions in early HCC patients (P<0.05);the accuracy and positive rate of serum miR-199a/b-3p detection in cirrhosis and HCC were better than AFP detection.Conclusion:Data indicated that miRNA-199a/b-3p had diagnostic value for indicating liver cirrhosis and early HCC.
3.Inhibition effect of interruption PAK4 expression on migration and invasion of hepatocellular carcinoma
Zihang WANG ; Chunshi LI ; Jinsong KANG ; Xuguang MI ; Lei LIU
Chinese Journal of Immunology 2015;(9):1183-1185
Objective:To analyze the effect of PAK4 interruption by microRNA-199a/b-3p (miR-199a/b-3p) on migration and invasion of hepatocellular carcinoma (HCC).Methods: To test targeting of PAK4 by miR-199a/b-3p,we used luciferase assay in HEK293T cells cotransfected miR-199a/b-3p mimcs and pmirGLO-PAK4 3′UTR.The expression of PAK4 in SMMC-7721 transfected with miR-199a/b-3p was detected by Western blot.The biology behaviors of SMMC-7721 cells transfected with miR-199a/b-3p or PAK4 Si were analysed by cell migration assay and invasion assay.Results:MiR-199a/b-3p could suppress the mRNA and protein ex-pression of PAK4 by targeting PAK4 3′UTR,and the downregulating PAK 4 expression suppress the migration and invasion of SMMC-7721 cells.Conclusion: MiR-199a/b-3p could suppress the expression of PAK 4, which are considered key HCC suppressors and inhibit the migration and invasion of HCC cells.
4.Effect of miR-199a/b-3p on cell motility of breast cancer cells
Zihang WANG ; Chunshi LI ; Jinsong KANG ; Xuguang MI ; Lei LIU
Chinese Journal of Immunology 2015;(9):1242-1244
Objective:To analyze the inhibiting mechanism of microRNA-199a/b-3p ( miR-199a/b-3p) on cell motility of breast cancer cells.Methods:The expression of PAK4 in MDA-MB-231 cells transfected with miR-199a/b-3p was detected by Western blot.The biology behaviors of MDA-MB-231 cells transfected with miR-199a/b-3p or PAK4 SiRNA were analysed by cell migration assay,invasion assay and protrusion dynamics.Results: MiR-199a/b-3p could suppress the expression of PAK 4 in MDA-MB-231 cells.Comparing with normal control ,miR-199a/b-3p or PAK4 SiRNA could suppress the migration ,invasion and membrane protrusion of MDA-MB-231 cells.Conclusion:miR-199a/b-3p could suppress the expression of PAK4,which are considered key breast cancer suppressors and inhibit the cell motility of breast cancer cells.
5.Relationship of clinical outcomes and expression of microRNA-199 a/b-3 p in HCC
Lei LIU ; Shouqing LI ; Xuguang MI ; Yanqiu FANG
Chinese Journal of Immunology 2015;(6):806-808
Objective:To investigate the expression of microRNA-199a/b-3p (miR-199a/b-3p) in hepatocellular carcinoma ( HCC) tissues,and to explore the relationship with clinical outcomes. Methods: Real time quantitative PCR technique was used to measure the expression of miRNA-199a/b-3p in HCC tissues. The correlation between miR-199a/b-3p expression and the clinic pathological features of patients were analyzed. Results: Comparing with adjacent control, miRNA-199a/b-3p presented lower expressions in HCC tissues (P<0. 05);lower miR-199a/b-3p was found correlated with metastasis and poor survival. Conclusion:MiR-199a/b-3p take a crucial role in HCC metastasis and recurrence.
6.Notch3 pathway mediates SAHA-induced apoptosis in human small-cell lung cancer H446 cells
Honglian CHEN ; Hui LIU ; Xuguang YANG ; Lei YUAN
Chinese Journal of Pathophysiology 2016;32(9):1556-1561
AIM: To investigate the effect of suberoylanilide hydroxamic acid (SAHA) on the apoptosis of hu-man small-cell lung cancer H446 cells and its possible mechanism.METHODS: H446 cells were incubated in the medi-um containing SAHA.CCK-8 assay was used to detect the anti-tumor effect of SAHA on the H446 cells, and IC50 values of SAHA were calculated.Flow cytometry was used to analyze the apoptosis.After Notch3 gene was silenced, the pro-apopto-tic effect of SAHA on the H446 cells was inhibited ( P <0.05).Eukaryotic expression plasmid containing N3ICD was transfected into the H446 cells, so that N3ICD was expressed in the H446 cells.The mRNA expression of Notch3 was measured by RT-PCR.The protein levels of Notch3, N3ICD, Puma and cleaved caspase-3 were determined by Western blot.RESULTS: SAHA remarkably reduced the cell viability in a dose-dependent manner (P <0.05), and the IC50 value of SAHA was 1.91 μmol/L.SAHA induced apoptosis in a dose-dependent manner (P <0.05).The expression of Notch3 gene was negative in the H446 cells, SAHA reactivated Notch3 gene and Notch3 pathway in a dose-dependent manner (P <0.05).Notch3 knockdown inhibited apoptosis induced by SAHA (P <0.05).Over-expression of N3ICD up-regula-ted the protein levels of Puma and cleaved caspase-3.CONCLUSION: SAHA induces apoptosis in human small-cell lung cancer H446 cells by activating Notch3 pathway and up-regulating the protein level of Puma.
7.Studies on the relationship of PDCD 4 with the invasion of astrocytic gliomas
Zhixian WAN ; Xiongwei WANG ; Yuanxun DONG ; Lei WANG ; Chunlei TIAN ; Xuguang WANG
Chongqing Medicine 2013;(32):3900-3901
Objective To investigate the relationship of programmed cell death 4(PDCD4) with the invasion of astrocytic glio-mas .Methods Using the immunohistochemical method to detect the expression of PDCD4 in astrocytic gliomas in different grades . Measuring the peritumoral low-density area on MRI scan ,then compared with the results of immunohistochemical expression .Re-sults The downregulation of PDCD4 was with the increasing of the malignant grade of astrocytic gliomas .The tumor grade malig-nancy was positively correlated with the grade of the peritumoral low-density area on MRI scan(P<0 .05) ,while the expression of PDCD4 was negatively correlated with the grade of astrocytic gliomas (P<0 .01) .Conclusion PDCD4 might serve as one of the in-dicators of invasion and malignant phenotype for astrocytic gliomas .
8.Preparation of SiO2/Ni core-shell type nanoparticles and their magnetic properties
Xinglong DONG ; Jinxuan LIU ; Hao HUANG ; Weina WANG ; Xuefeng ZHANG ; Junpeng LEI ; Xuguang ZHU
Chinese Journal of Tissue Engineering Research 2008;12(27):5397-5400
BACKGROUND: Inhibitory coating can prevent nanoparticle oxidation, grain growth, corrosion and agglomeration, and endow nanoparticle with special properties. ABJECTIVE: To prepare SiO2/Ni core-shell type nanoparticles and assess their magnetic properties. DESIGN, TIME AND SETTING: The observation experiment was performed between November 2005 and March 2006 at Nanometer Compound Material Research Laboratory of Dalian University of Technology, Dalian, Liaoning Province, China. MATERIALS: Nanometer nickel powder prepared by DC arc plasma jet method, Na2SiO3 produced by Bazhou Chemical Industry Branch Factory of Tianjin Quartz Clock Factory (China).METHODS: SiO2/Ni core-shell type nanoparticles were synthesized by coating a layer of SiO2 on the surface of manometer nickel powder via liquid deposition method using Na2SiO3 as the main source material. MAIN OUTCOME MEASURES: Their microstructures and material properties were investigated by X-ray diffraction, Fourier transform infrared spectrometer, transmission electron microscopy, thermo-gravimetric analysis, differential scanning calorimetry and vibrating sample magnetometer. RESULTS: The experimental results showed that SiO2 shell was in an amorphous state around Ni cores and it avoided agglomeration of the Ni nanoparticles. The oxidation temperature of nanometer nickel powder coated by SiO2 elevated from 287 ℃ to 385 ℃. The analysis result of magnetic properties indicated that the hysteresis loop of Ni had an excursion for the existence of anti-ferromagnetic NiO, the silica coating reduced the saturation magnetization and improved the coercivity. CONCLUSION: Preparation of SiO2/Ni core-shell type nanopartieles was successful; silica coating improved the oxidation resistance of nanometer nickel powder, endowed nanometer nickel powder better ferromagnetism and improved the coercivity.
9.Correlation analysis between expression of NF-κB and quartz's deposit in non-small cell lung cancer in Xuanwei , Yunnan province
Mengjiao QIAN ; Yunchao HUANG ; Xuguang PENG ; Guangjian LI ; Guangqiang ZHAO ; Linwei TIAN ; Lianhua YE ; Yujie LEI
Cancer Research and Clinic 2017;29(8):551-555
Objective To evaluate the relationship between quartz's deposit and expression of NF-κB in non-small cell lung cancer (NSCLC) lung tissues in Xuanwei, Yunnan Province, and to clarify the role of quartz in Xuanwei NSCLC's carcinogenic mechanism. Methods As research objects, the lung tissues of NSCLC and lung benign lesions after surgical resection were collected from July 2009 to September 2015 at the Third Affiliated Hospital of Kunming Medical University. Firstly, the transmission electron microscopic (TEM) with energy dispersive X-ray analyzer (EDS) was used for observation of crystalline deposit and local pathological changes. Secondly, expression level of NF-κB had been analysed and a correlation analysis with particle size of SiO2 crystal in the same lung sample was made. Results The occurrence rates of quartz in Xuanwei NSCLC lung tissues were above non-Xuanwei NSCLC and benign lung tissues (P<0.01);the average particle size of SiO2 crystal was (226 ± 120) nm × (237 ± 163) nm in Xuanwei NSCLC group and it was smaller than the other two groups; In Xuanwei NSCLC group, the expression level of NF-κB was significantly higher than non-Xuanwei and benign lung tissues (P< 0.01), but there was no significant difference between cancer tissues and normal lung tissues in the group (P>0.05). The expression level of NF-κB was generally increasing when quartz 's size became smaller. Conclusion Quartz 's deposit may play a certain role in carcinogenic mechanism of lung cancer in Xuanwei, the smaller the particle size, the greater the cytotoxicity.
10.Influence of tannic acid on contents of 8-OHdG and 3-NT in nephridial tissue of diabetic rats and supernatant of glomerular mesangial cells
Haifeng WEI ; Lei LIU ; Xiaodan LU ; Xuguang MI ; Shouqing LI ; Yan TAN ; Yanqiu FANG
Chinese Journal of Immunology 2014;(8):1040-1045
Objective:To observe the effect of tannic acid on renal morphology and function of diabetes mellitus model rats ,and to explore the mechanism of improving effect of tannic acid from oxidative stress , nitrosative stress angle.Methods: 8 rats were randomly selected from 68 6-week-old male Wistar rats as normal control group and the remaining 60 rats accepted high-sugar and high-fat diet for 4 weeks, then were injected streptozotocin ( STZ, 52 mg/kg ) intraperitoneally in order to manufacture a diabetic rat model.Further the diabetic rats were randomly divided into model group ,aminoguanidine group ,low-dose of tannic acid group and high dose of tannic acid group.The rats in aminoguanidine group were injected aminoguanidine [AG,40 mg/(kg· d)] intraperitoneally, those in low-dose of tannic acid group were injected tannic acid [TA,20 mg/(kg· d)] and those in high-dose of tannic acid group were injected tannic acid [TA,30 mg/(kg· d)].The rats in normal control group and model group were injected normal saline [NS, 30 mg/(kg· d)] and all rats were sacrificed and tissues were derived at the end of the week 10.Morphologic changes of kidney in diabetic rats were observed by HE staining and correlative biochemical indices of renal function were detected by biochemical analyzer.8-hydroxy deoxyguanosine (8-OHdG) and 3-nitrotyrosine (3-NT) content of renal tissue in rats in different groups were detected by ELISA method.Mesangial cells cultured in vitro were treated with high concentration of glucose (30 mmol/L) and AGEs (250 mg/L) and at the same time with different concentration of tannic acid (10,20,40 and 80μmol/L) on the basis of setting corre-sponding control group.The contents of 8-OHdG and 3-NT in the culture supernatant were measured by ELISA method after 48 hours.Results:Tannic acid can effectively improve the renal pathological changes and improve renal function of diabetic rats .The contents of 8-OHdG and 3-NT in kidney tissue homogenate of diabetic rats and in the supernatant of GMC cultured with high glucose or AGEs were all significantly increased and can be reduced by tannic acid.Conclusion:Tannic acid improving the structure and function damage of kidney in diabetic rats might be achieved by oxidative stress and nitrosative stress mechanism .