1.Effect of nano-aIumina on mitophagy in primary corticaI neuronaI ceIIs from Wistar newborn rats
Lijun CHANG ; Weiwei GUO ; Cuicui GE ; Qiao NLU ; Qinli ZHANG
Chinese Journal of Pharmacology and Toxicology 2014;(5):737-742
OBJECTIVE To study the effect of nano-alumina(nano-Al2 O3 )on mitophagy in primary cortical neuronal cells from Wistar newborn rats. METHODS The purity of neuronal cells was detected by immunohistochemistry,and the lactate dehydrogenase(LDH)assay was performed to determine the viability of the cells treated with 13 nm nano-Al2 O3 0.5 mmol·L-1 for 12,24 and 48 h,respectively. The mitochondrial membrane potential(MMP)was detected by flow cytometry analysis . The ultrastructure of mitochondria and mitophagy vacuoles was observed by transmission electron microscopy(TEM). Auto-phagic vacuoles were observed by dansylpentanediamine(MDC)staining and the expression of autoph-agy related protein Beclin1 and LC3Ⅱ/ Ⅰ was determined by Western blotting. Mitophagy was observed by Lysotracker and Mitotracker staining respectively. RESULTS More than 95% cells were neuronal cells. The activity of LDH in the supernatant liquid exposed to nano-Al2 O3 for 12 and 24 h groups was sig-nificantly increased compared with the control group(P﹤0.05). After exposure to nano-Al2 O3 ,the mito-chondrial membrane potential was significantly decreased compared with the control group( P ﹤0.01). The results of TEM displayed mitochondrial swelling and the formation of vacuoles and mitophagy in nano-Al2 O3 groups. MDC positive fluorescence particles were observed and the expression of autophagy related protein Beclin1 and LC3Ⅱ/ Ⅰ was increased in nano-Al2 O3 groups compared with the control group( P ﹤ 0. 05 ). The result of Lysotracker and Mitotracker colocalization showed the fusion of mitochondria and lysosomals. CONCULSION Nano-Al2 O3 may induce autophagy and mitochondria damage in neuronal cells while the damaged mitochondria may be removed by mitophagy.
2.NF-κBp50 is Associated With DC-SIGN Expression Induced by IL-4 in THP-1 Cells
Lijun XU ; Xiuchun CHANG ; Hangping YAO ; Nanping WU
Progress in Biochemistry and Biophysics 2008;35(1):50-55
DC-specific intercellular adhesion molecule-3-grabbing nonintegrin (DC-SIGN) is specific receptor on Dendritic cells, and plays a pivotal role on antigens presentation. Uptodate, the clear regulation mechanisms for DC-SIGN expression are not available.IL-4 is one of the most important cytokines inducing DC-SIGN production, while, NF-κB is an important transcription factor controlling signaling transduction. Both IL-4 and NF-κB are closely related to DC-SIGN regulation. NF-κB and IL-4 actions on DC-SIGN promoter activity, DC-SIGN expression as well as interactions between IL-4 and NF-κB were investigated in THP-1 cell. It was found that the mutation of NF-κB binding site in DC-SIGN promoter results in DC-SIGN promoter activity decrease about 50%.NF-κBp50 stimulates DC-SIGN expression in THP-1 cells. IL-4 upregulates DC-SIGN expression on THP-1 cells as well as NF-κB production. These data reveal that NF-κB is associated with IL-4 induced DC-SIGN expression.
3.Mechanism of the mitochondria damage of the nerve cells in mouse after chronic exposure to nano-alumina
Weiwei GUO ; Lijun CHANG ; Yong DING ; Huan LI ; Cuicui GE ; Haiyang WANG ; Qinli ZHANG ; Qiao NIU
Chinese Journal of Pharmacology and Toxicology 2014;(2):194-198
OBJECTIVE To explore the potential neurotoxicity of nano-alu mina (<50 n m)in vivo, we treated the ICR mouse with the nano-alu mina to investigate the mitochondrial da mage of nerve cells on morphology and function.METHODS Adult male mice were exposed to nano-alu mina (<50 n m)of 0,25,50 and 75 mg·kg -1 by nasal instillation for 1 month.Then we observed the mitochondrial ultra-structure of the nerve cells in CA3 region of hippoca mpus,and measured the mean dia meter in every group.The activities of Na +-K +-ATPase and Ca2 +-Mg2 +-ATPase were tested by the determination of the inorganic phosphorus,which was the deco mposition product of ATPase.Western blot analysis was used to detect the expression of COX-Ⅳ,Beclin1 ,LC3Ιand LC3Ⅱ.RESULTS Co mpared with 0 and 25 mg·kg -1 groups exposed to Al2 O3 nanopartilces (Al2 O3 NPs),the mitochondria of CA3 region in hip-poca mpus in 50 mg·kg -1 group beca me ede matous and swollen with sparse and broken cristae sur-rounding the nuclear,and the mean dia meter was higher(0.49 ±0.02 μm,P <0.05).But co mpared with 50 mg·kg -1 group,the mitochondria in 75 mg·kg -1 group beca me s maller with inner cristae of high density,and the mean dia meter was lower(0.36 ±0.02 μm,P<0.05).The enzy me activity of the mito-chondria in cerebral cortex decreased dose-dependently with exposure,the activities of Na +-K +-ATPase in 50 and 75 mg·kg -1 groups(6.37 ±0.22 kU·g -1 protein,5.48 ±1 .53 kU·g -1 protein)and Ca2 +-Mg2 +-ATPase in 50 and 75 mg·kg -1 groups (3.21 ±0.99 kU·g -1 protein,3.28 ±0.15 kU·g -1 protein)were lower than the 0 mg·kg -1 group(P<0.05).Meanwhile,the Ca2 +-Mg2 +-ATPase in 50 and 75 mg·kg -1 groups showed lower activities in co mparison with the 25 mg·kg -1 group.The 75 mg·kg -1 group expressed higher level of the COX-Ⅳ protein 1 .35 ±0.66(P<0.05)than other groups.Both expression of Beclin1 protein and rate of LC3Ⅱ/LC3Ⅰin 75 mg·kg -1 group were more than the 0 mg·kg -1 group. CONCLUSION The mitochondrial dysfunction may be the potential neurotoxicity of nano-alu mina,and the da maged mitochondria were cleared by autophagy.
4.Megavoltage cone-beam CT in the evaluation of set-up errors in head and neck cancers treated with precision radiotherapy
Yunsheng GAO ; Xi CHANG ; Lijun ZHOU ; Weigang HU ; Xiaoshen WANG ; Guopei ZHU ; Yongru WU ; Chaosu HU
Chinese Journal of Radiation Oncology 2010;19(3):263-266
Objective To evaluate set-up errors by megavoltage cone-beam CT in head and neck cancers treated with precision radiotherapy. Methods From April 2007 to March 2008, 22 patients with nasopharyngeal carcinoma (15 patients) ,parotid carcinoma (4 patients) and brain glioma (3 patients) were enrolled, among whom 7 patients underwent three-dimensional conformal radiotherapy (3DCRT) and 15 received intensity modulated radiotherapy (IMRT). The radiation dose was 56. 0 -70.4 Gy in 28 -32 fractions within 6 -7 weeks. Megavoltage cone-beam scan was performed weekly before treatment. The isocenter displacement was calculated by comparing megavoltage cone-beam CT and planning CT in left-right (LR), cranio-caudal (CC), and anterior-posterior (AP) directions. Results Totally 129 sets of megavoltage cone-beam CT images were obtained for 22 patients. The frequency of isocenter shift more than 0. 3 cm,0. 4 cm and 0. 5 cm were 28,15 and 9, respectively. The maximum mean set-up error was found in CC, which was about 0. 1 cm more than that in LR and AP. The frequency of isocenter shift direction was almost identical in LR and CC, which was more frequent (about 75%) in the posterior direction. Conclusions During the course of radiation of brain tumor and head and neck cancer, the enlarging tendency of set-up errors has been found in all the three directions. The isocenter shift in AP was more frequent to the posterior direction.
5.Isolating method of rat pancreatic islets and affect factors for its activity
Baocheng CHANG ; Hong ZHANG ; Ning ZHENG ; Wei ZHAO ; Qiumei ZHANG ; Lijun WEI ; Peihua FANG
Basic & Clinical Medicine 2006;0(09):-
Objective To assess the factors which affect the activity of isolated islets,and to construct a steady and effective isolation method of rat pancreatic islets.Methods Pancreatic islets were isolated by collagenase method.The glucose-stimulation of insulin secretion(GSIS) in different conditions was performed by batch incubation method and measured by RIA.Results GSIS was improved by BSA;There was a evident decrease of insulin secretion in cultured islets of 7 days,however,no difference was observed between the freshly isolated islets group and the 1~5days cultured islets groups.Compared to the groups of(5.5 mmol/L) and(25 mmol/L) glucose in the culture medium,(11.1 mmol/L) glucose group stimulated higher insulin release.Conclusion BSA,glucose concentration of RPMI1640 medium and the culture period are related to the activity of isolated islets.
6.Association of non-HLA gene polymorphisms with CMV infection after hematopoietic stem cell transplantation
Ziling ZHU ; Xiaojin WU ; Yufeng FENG ; Lijun DAI ; Huirong CHANG ; Depei WU
Chinese Journal of Organ Transplantation 2013;34(7):419-423
Objective To explore the non-HLA gene polymorphisms that influence CMV infection after hematopoietic stem cell transplantation (HSCT).Method Non-HLA gene (ACE,CD14,MPO,MBL) single nucleotide polymorphisms were determined by using sequence-specific primer polymerase chain reaction (PCR-SSP) and sequencing in 64 pairs of donors and recipients before HSCT and the differences of non-HLA gene were analysed in CMV positive and negative patients Results The distribution of ACE gene single nucleotide polymorphism was DD (14/128,10.9%),ID (72/128,56.3%),and Ⅱ (42/128,38.8%).The distribution of CD14-159 allele gene single nucleotide polymorphism was CC (18/128,14.1%),CT (81/128,63.3%),and TT (29/128,22.7%).The distribution of MPO-463 allele gene single nucleotide polyrnorphism was G (100)/128,78.1%),A (2/128,1.6%),and GA (26/128,20.3%).The distribution of MBL gene single nucleotide polymorphism was H (28/128,21.9%),HL (73/128,57.0%),L (27/128,21.1%),Y (87/128,68.0%),YX (38/128,29.7%),X (3/128,2.3%),A (94/128,73.4%),AB (32/ 128,25.0%),and B (2/128,1.6%).The allele frequency of ACE,CD14 and MPO shoed no significant differcence between CMV positive and negative patients The gene frequency of MBL-HL was increased in CMV positive group.Conclusion MBL gene single nucleotide polymorphisrns may influence CMV infection after HSCT.
7.Changes and clinical relevance of plasma interleukin-6 and high sensitive C-reactive protein in patients with coronary heart disease and angina
Lijun LIU ; Yingxia Lü ; Shuanli XIN ; Chao CHANG ; Shouli WANG ; Xiufeng ZHAO
Clinical Medicine of China 2010;26(12):1274-1276
Objective To explore the roles of interleukin-6 (IL-6) and high sensitive C-reactive protein (hsCRP) in coronary heart disease (CHD). Methods One hundred and fifty patients diagnosed as angina pectoris by coronary angiography were enrolled and randomly divided into unstable angina pectoris group ( UAP group n =78) and stable angina group (SAP group n =72). At the same time,50 cases with normal coronary angiography were selected as control. The plasma level of IL-6 was measured by ELISA, and hsCRP was measured by immunonephelometry. Results The mean plasma levels of IL-6 and hsCRP were significantly higher in UAP group compared with SAP group ( IL-6 ( 36. 98 ± 9. 35 ) pg/L vs. ( 23.19 ± 7. 35 ) pg/L; hsCRP ( 2. 25 ± 0. 73 ) μg/Lvs 1.88 ± 0. 68 ) μg/L; P < 0. 01 respectively), both were significantly higher than those of control group ( IL-6 (16. 93 ± 6. 15 )pg/L and hsCRP (1.28 ± 0.40)μg/L,P <0.01 respectively). We also found significantly positive relationship between IL-6 and hsCRP plasma level ( r = 0. 875, P < 0. 05 ). Conclusions Inflammatory response might play an important role in CHD. IL-6 and hsCRP were of high value in prediction of angina pectoris and development of CHD.
8.Effect of plasma monocyte chemotactic factor-1 protein and matrix metalloproteinase-9 of patients with coronary artery disease following percutaneous coronary interventional procedures therapy
Qin LI ; Lijun LIU ; Shuanli XIN ; Chao CHANG ; Dongyan LIU ; Xuexia CUI ; Shuli ZHANG ; Yaqi LI ; Yanjun DAI ; Fuxian ZHANG
Clinical Medicine of China 2014;(7):698-700
Objective To explore the change of monocyte chemotactic factor-1 protein(MCP-1)and matrix metalloproteinase-9( MMP-9)of patients with coronary artery disease( CAD)following percutaneous coronary interventional( PCI). Methods Fifty patients underwent PCI procedures for CAD compromising a single coronary artery were selected as PCI group and 30 healthy individuals with normal findings by coronary angiography were selected as the control group. Plasma MCP-1 and MMP-9 were measured in all the subjects. Results The plasma MCP-1 level of patients with CAD after PCI was(19. 87 ± 5. 31)ng/ L,higher than that before operation((15. 71 ± 5. 23)ng/ L,t = 3. 95,P < 0. 01). Whereas in the control group,the MCP-1 level after coronary angiography was(13. 78 ± 5. 58)ng/ L,which was as same as that before operation (12. 42 ± 5. 39 ng/ L,P = 0. 34). Plasma MMP-9 level in the CAD patients after PCI procedures was(22. 69 ± 5. 97)mg/ L,higher than that before operation((19. 52 ± 5. 72)mg/ L,t = 2. 71,P < 0. 01). There was no significant difference in term of plasma MMP-9 level in control group befor and after operation((17. 53 ± 5. 51) mg/ L vs.(16. 69 ± 5. 42)mg/ L,P = 0. 55). Conclusion Plasma MCP-1 and MMP-9 increase in CAD patients following PCI procedures. But their roles in the vascular restenosis following the procedures need further investigation.
9.Influence of rhBNP on cardiac function,levels of hsCRP and MMP-9 in patients with acute heart fail-ure
Min LI ; Chao CHANG ; Shuanli XIN ; Jixiang LIU ; Lijun LIU ; Liying HAN ; Xiufeng ZHAO ; Renjie ZHANG ; Zhijiang XIE
Chinese Journal of cardiovascular Rehabilitation Medicine 2015;24(4):415-417
Objective:To evaluate the clinical therapeutic effect and mechanism of recombinant human brain natri-uretic peptide (rhBNP,Xinhuosu)in patients with acute heart failure (AHF).Methods:A total of 90 AHF patients were randomly and equally divided into rhBNP group (received intravenous injection of rhBNP based on routine treatment for 3d)and routine treatment group.Left ventricular ejection fraction (LVEF),levels of serum high sen-sitive C reactive protein (hsCRP)and matrix metalloproteinase-9 (MMP-9)were measured and compared between two groups before and after treatment.Results:Compared with before treatment,after treatment,LVEF signifi-cantly rose,levels of hsCRP and MMP-9 significantly reduced in both groups,P <0.05 or <0.01;compared with routine treatment group after treatment,there was significant rise in LVEF [(41.4±12.8)% vs.(51.3±13.9)%], and significant reductions in levels of hsCRP [(8.63±3.57)mg/L vs.(6.67±3.97)mg/L]and MMP-9 [(17.89 ±4.75)mg/L vs.(14.64±4.89)mg/L]in rhBNP group,P <0.05 or <0.01. Conclusion:The rhBNP possesses significant therapeutic effect on acute heart failure,and its mechanism may be related with reducing hsCRP and MMP-9 levels.
10.Dose characteristics of megavoltage cone-beam CT in the low MU and rotation way for imaging
Zhiyong XU ; Longgen LI ; Lijun ZHOU ; Jianfian QW ; Xi CHANG ; Weigang HU ; Junchao CHEN ; Zhen ZHANG ; Guoliang JIANG
Chinese Journal of Radiation Oncology 2008;17(2):120-122
Objective To investigate dose characteristics of megavoltage cone-beam CT in the low MU and rotation way for imaging,and to improve the quality assuraflce. Methods Dose linearity,a parameter describing beam characteristics,was measured for different numbers of MUs on megavoltage conebeam CT accelerator.Dose stability was measured 5 times using exposure of 7 MUs,which was repeated another 5 times 6 hours after. Results Dose linearity was good using exposure between 5 and 20 MUs.The average dose among the first and second 5 measurements was(5.74±0.01)Gy and(5.76±0.01)Gy while exposure of the accelerator was 7 MU.The difference between the two doses was 0.41%. he dose was stability while megavoltage cone-beam CT image was in the low MU and rotation way. Conclusion Dose characteristics of megavoltage cone-beam CT in the low MU and rotation way for imaging can qualify clinically application.