1.Effect of omega-3 polyunsaturated fatty acid on lipopolysaccharide-induced acute lung injury in rats
Yan ZHAO ; Xinxin LIU ; Dan GUO ; Lei WANG ; Ping CHEN
Chinese Journal of Clinical Nutrition 2013;(2):83-89
Objective To evaluate the effect of dietary ω-3 polyunsaturated fatty acid (PUFA) supplementation on lipopolysaccharide (LPS)-induced acute lung injury in rats.Methods Totally 58 male SD rats were divided into control group (n =10),model group (n =12),ω-3 PUFA high-dose group (n =12),ω-3PUFA medium-dose group (n =12),and ω-3 PUFA low-dose group (n =12).Seven days before model establishment,rats in the three ω-3 PUFA groups were orally given ω-3 PUFA at 1,0.5,and 0.25 g/kg body weight once per day,respectively,for seven consecutive days.Twenty-four hours after the last administration,all rats except those in the control group were given intravenous injection of LPS (6 mg/kg) at caudal vein to establish the model of acute lung injury.Body temperature was measured at 0,6,and 24 hour.Blood samples were collected from the eye venous plexus for routine blood tests and blood biochemical tests 24 hours after modeling.After the rats were sacrificed,the left lung was harvested for measuring the wet weight and dry weight and calculating the wet/dry weight ratio (W/D).The right lung was harvested for pathological observation under light microscope and calculation of semi-quantitative pathological index (PI).Results Twenty-four hours after modeling,deaths were noted in all groups except the control group.After injection of LPS,rats curled with little movements.At 6 hour,the body temperature was significantly higher in the model group than in the control group [(37.4 ±0.27)℃ vs.(35.9 ±0.05) ℃,P =0.00] ; it was (36.2 ±0.38)℃,(36.3 ±0.30)℃,and (36.3 ± 0.32) ℃ in the ω-3 PUFA high-,medium-,and low-dose groups,which were significantly lower than that in the model group (all P =0.01).The amounts of white blood cells,neutrophils,and lymphocytes increased in the model group,but showing no significant difference compared with the other groups.The serum glutamic oxalacetic transaminase (GOT) and glutamic pyruvic transaminase (GPT) levels were significantly higher in the model group than in the control group [(353 ± 235) U/L vs.(157 ± 55) U/L,P =0.02 ; (141± 103) U/L vs.(54 ±23) U/L,P =0.03] ; the ω-3 PUFA high-dose group had significantly lower GOT and GPT levels than the model group did [(167 ±94) U/L vs.(353 ±235) U/L,P =0.03 ; (63 ±57) U/L vs.(141 ± 103) U/L,P =0.04].The model group had significantly higher lung wet weight [(371 ±38) mg vs.(281 ±24) mg,P=0.01] and W/D value (7.34±1.40 vs.5.41 ±0.84,P=0.01) compared with the control group.Compared with the model group,the W/D value was significantly lower in the ω-3 PUFA high-,medium-,and low-dose groups (6.17 ±0.58,P =0.03; 6.17 ± 0.76,P =0.03; 6.13 ± 1.23,P =0.04).Light microscopy showed that the lung alveoli of the model group presented congestion,obvious expansion,and scattered inflammatory cell infiltration in interstitium,along with significantly increased PI compared with the control group (3.9±0.9 vs.0.0±0.0,P=0.00).The PI value was (2.1 ±0.3),(2.1 ±0.3),and (2.3 ± 0.5) in ω-3 PUFA high-,medium-,and low-dose groups,respectively,all significantly lower than that in the model group (all P =0.01).Conclusions The acute lung injury model could be successful established by intravenous injection of LPS.ω-3 PUFA at different doses can improve the acute lung injury of rats.It is therefore supposed that early enteral administration of ω-3 PUFA can alleviate LPS-induced acute lung injury,although the optimal dosage and timing need further research.
2.Glucagon-like peptide-1-induced myoblast line C_2C_(12) cells differentiates into cells secreting insulin-like substance
Chuan YANG ; Hua CHENG ; Dan LIU ; Lei ZHU
Chinese Journal of Endocrinology and Metabolism 2000;0(06):-
Myoblast line C_2C_ 12 cells were co-cultured with various concentrations of glucagon-like peptide-1 (GLP-1). Being induced by GLP-1, myoblast line C_2C_ 12 cells have the potential to differentiate into cells which are able to secrete insulin-like substance.
3.PURIFICATION AND PROPERTIES OF SECRETORY ASPARTIC PROTEINASE FROM CANDIDA ALBICANS
Dan-Min WANG ; Li-Yin LIU ; Lu YE ; Lei GAO ;
Microbiology 1992;0(02):-
An extracellular proteinase from Candida albicans WD27 was purified by (NH4)2SO4 fractionation and DEAE-Sephacel ion-exchange chromatography with 25.4 fold and 5.2% yield. This enzyme appeared to be aspartic proteinase since the enzyme activity could be inhibited by pepstatin which was specific inhibitor of this class of proteinase. The enzyme had an acidic proteolytic activity profile with the optimum pH of 4.0. The optimum temperature of the enzyme activity was 37℃. The proteinase had a broad substrate specificity with the highest susceptibility to bovine hemoglobin. The Km for bovine hemoglobin was determined to be 0.814mmol/L.
5.Multiple center study about the reliability of the low-dose dual-source CT coronary angiography in the step-and-shoot mode
Tao BI ; Lei XU ; Zhaoqi ZHANG ; Lin YANG ; Lei ZHAO ; Bin Lü ; Dan HAN ; Cheng LIU ; Qing DUAN
Chinese Journal of Radiology 2009;43(7):708-713
Objective To investigate the accuracy of low-dose dual-source computed tomography (DSCT) coronary angiography in the step-and-shoot (SAS) mode for the diagnosis of coronary artery stenosis in comparison with conventional coronary angingraphy (CCA).Methods Prospective multiple-center study, 46 patients[mean age(58±9) years;bedy mass index(BMI) (25±3) kg/m2]underwent both DSCT in the SAS mode and CCA within 14 days.The inclusion criteria for contrast-enhanced CT: (1) heart rate less than 65 times/rain (bpm).(2) regular sinus rhythm, heart rate fluctuations within the range of 6 bpm. (3) holding breath well, breath-hold time is about 12-15 s.The exclusion criteria:(1) allergy to iodinecontaining contrast medium, nephropathy (serum creatinine level 120 μmol/L), heart failure and serious arrhythmias.(2) patients with coronary stents or bypass grafts.(3) heart rate can not be controlled very well (4)the patient could not take nitroglycerin.(5)BMI 30 kg/m2.(6) other heart disease: carcliomyopathy, valvular disease etc.Sensitivity, specificity, negative (NPV) and positive predictive value (PPV) were determined with CCA as standard of reference.The Kappa value between the two modalities and the two observers was calculated.Radiation dose values were measured.Results Mean heart rate during scanning was (61±6)bpm.99.19% (614/619) coronary segments were depicted with a diagnostic image quality. The vessel-based sensitivity, specificity, PPV, and NPV for the diagnosis of coronary artery stenosis were 96.2% (75/78), 88.2% (60/68), 90.4% (75/83), and 95.2% (60/63), respectively.The Kappa value between the two modalities was 0.848 (P=0.000).The mean effective dose of the SAS-CTCA was (2.95± 0.96) rosy(1.26-4.32 mSy).Conclusion In selected patients, DSCT coronary angiography in the SAS mode have good image quality, which allows for the accurate diagnosis of coronary stenosis at a low radiation dose.
6.Effects of preconditioning and postconditioning with shenfu injection on cognitive function in patients after valve replacement under cardiopulmonary bypass.
Lei ZOU ; Dan-Yan LIU ; Yu CAO
Chinese Journal of Integrated Traditional and Western Medicine 2009;29(6):497-500
OBJECTIVETo investigate the effects of preconditioning and postconditioning with Shenfu Injection (SFI) on cognitive function in patients after valve replacement under extra-corporeal circulation.
METHODSThirty-two patients prepared to receive valve replacement, aged 25-54 years, with heart function of II-III level, were randomly assigned to four groups, eight in each group. Patients in group E1 received SFI 1 mL/kg after intubation and before blocking the aorta; patients in group E2 received SFI 1 mL/kg after opening the aorta; patients in group E3 received SFI 0.5 mL/kg twice, at before blocking and after opening the aorta, respectively; and patients in group C received 1 mL/kg normal saline after intubation for control. All the medication was infused via pump. Venous blood samples were taken from the internal jugular venous bulb cannula for detecting plasma S100beta protein by ELISA at 6 different time points, i.e. after trachea intubation (T1), 10 min after cardiopulmonary bypass (CPB, T2), hypothermia stabilizing stage (T3), re-warming to 33 degrees C (T4), ending CPB (T5) and 1 h after ending CPB (T6). And patients' cognitive function was assessed for 4 times with mini-mental state examination (MMSE) scale, at the day before operation, and 1, 2, 7 days after operation.
RESULTSThe elevation of S100beta plasma protein was lesser in the three E groups than that in group C (P < 0.05), and the lowest level was shown at T6 in Group E3 (P < 0.05). The highest incidence of cognitive dysfunction occurred in Group C one week after operation (P < 0.05).
CONCLUSIONSFI may reduce the plasma level of S100B protein, maintain stable the structure and function of blood-brain barrier, it is favorable to the post-operational recovery of neurological function of patients, showing good brain protective effect. The optimal effect could be obtained by pump infusion of 0.5 mL/kg of SFI before aortic blocking and after aortic opening.
Adult ; Blood-Brain Barrier ; drug effects ; Brain ; blood supply ; Cardiopulmonary Bypass ; Cognition Disorders ; prevention & control ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Heart Valve Prosthesis Implantation ; Humans ; Ischemic Preconditioning ; Male ; Middle Aged ; Nerve Growth Factors ; blood ; Phytotherapy ; Postoperative Complications ; prevention & control ; S100 Calcium Binding Protein beta Subunit ; S100 Proteins ; blood
7.Expression profiles of genes in wild-type DJ-1 and A39S DJ-1 mutant cells
Zhenhua LIU ; Beisha TANG ; Dan HE ; Lei WANG ; Zhiquan XIAO ; Xinxiang YAN ; Jifeng GUO
Journal of Central South University(Medical Sciences) 2014;(4):325-332
Objective: To elucidate the role of A39S mutation of DJ-1 in the onset of Parkinson’s disease (PD) and identify genes for which expressions are abnormally regulated by A39S DJ-1 mutation. Methods: We established HEK293 cell lines which stably expressed empty vector, wild-type DJ-1 and A39S mutated DJ-1 respectively. DNA microarrays were used to identify genes for which expressions change in wild-type DJ-1 cells and A39S DJ-1 mutant cells. Results: Compared with the cell line expression empty vector, we identified 42 differentially regulated genes (including 14 up-regulated genes and 28 down-regulated genes) in the wild-type DJ-1 cells and 8 differentially regulated genes (including 6 up-regulated genes and 2 down-regulated genes) in the A39S DJ-1 mutant cells. Compared with the wild-type DJ-1 cells, only the expression of UGT2B7 gene was down-regulated in A39S DJ-1 mutant cells. hTese differentially regulated genes were mainly related to signal transduction, regulation of transcription, apoptosis and metabolism. Conclusion: A39S mutated DJ-1 may disturb the transcriptional activities of DJ-l and involve in the pathogenesis of PD.
8.Tumor specific immune response induced by three tandem repeats of CEA mini gene DNA vaccine and safety evaluation
Yanqiu FANG ; Haifeng WEI ; Dan LI ; Xuguang MI ; Xiaodan LU ; Shouqing LI ; Lei LIU ; Yan TAN
Chinese Journal of Immunology 2017;33(2):217-219,225
Objective:To observe the specific killing effect on tumor cells of the spleen cells in mice immunized with three tandem repeats of CEA minigene DNA vaccine pcDNA-triCEA625-667 and to evaluate the safety of the vaccine. Methods: The BALB/c mice were randomly divided into blank vector group ( pcDNA3. 0 ) , haploid vaccine group ( pcDNA-CEA625-667 ) and tandem repeats vaccine group (pcDNA-triCEA625-667). The mice received a total of 4 intramuscular immunization every 10 days once. The changes of body weight,survival state were recorded and the levels of serum ALT and serum creatinine were detected. The specific CTL killing activity of spleen cells in accinated mice on mouse hepatoma cells(H22-CEA+),gastric cancer cells(MFC-CEA+),colorectal cancer cells ( CT26-CEA+) with high expression of CEA and mouse hepatoma cells ( H22-CEA-) without expression of CEA was detected. Results:The two vaccines had strong killing activity on CEA positive liver cancer,gastric cancer and colon cancer cells,and the difference was statistically significant ( P<0. 01 ) compared with the PcDNA3. 0 group. And they had almost no effect on CEA negative tumor cells (H22-CEA-). The killing activity on liver cancer cell(H22-CEA+) and gastric cancer cell(MFC-CEA+) induced by pcDNA-triCEA625-667 was stronger than that induced by pcDNA-triCEA625-667(P<0. 05). The survival status,change of body weight and function of liver and kidney of the mice were not affected by the vaccine. Conclusion:There was no adverse reaction in the course of vaccine immunization. The minigene DNA vaccine derived from CEA can induce tumor specific CTL effect and the immune response level elicited by three tandem repeats of minigene DNA vaccine was superior to that elicited by haploid vaccine.
9.Role of Bcl-2 signal pathway in apigenin preconditioning against cardiomyocytes anoxia/reoxygenation injury
Min ZHOU ; Jiazhen YOU ; Huan HE ; Dan LIU ; Zhangping LIAO ; Lei TANG ; Dong YIN ; Ming HE
Chinese Pharmacological Bulletin 2015;(1):122-126,127
Aim To investigate the relationship be-tween the cardioprotection of apigenin ( Api ) from an-oxia/reoxygenation ( A/R) injury and Bcl-2 pathway. Methods H9 c2 cardiomyocytes were cultured and di-vided into normal control group, A/R group, Api pre-treatment group ( Api ) , Api + Bcl-2 inhibitor group ( Api + ABT-737 ) . Expression of Bcl-2 was deter-mined by Western blot,and cell viability was measured by MTT method. LDH, SOD, GSH-Px, MDA activity were determined by chromometry. ROS generation, mi-tochondrial membrane potential and apoptosis were de-termined by flow cytometry. Results 25h after apige-nin precondition,the expression of Bcl-2 was upregulat-ed in cardiomyocytes ( P <0. 01 ) . In the group pre-treated with 40 μmol · L-1 apigenin before A/R, the activity of LDH in culture medium decreased; the ac-tivity of intracellular SOD, GSH-Px increased; the content of MDA and ROS generation decreased; cell viability increased; mitochondrial membrane potential could be more stable and cell apoptosis decreased ( P<0. 01 ) . However, all these protective effects were attenuated significantly in the group pretreated with apigenin and Bcl-2 inhibitor ABT-737 . Conclusion The effect of apigenin against A/R injury in cardiomyo-cytes involves Bcl-2 pathway, and at least partly de-pends on its effect on upregulating the expression of Bcl-2 .
10.Observation of anti tumor activity in vivo of three tandem repeats of minigene DNA vaccine derived from CEA gene
Yanqiu FANG ; Haifeng WEI ; Dan LI ; Xuguang MI ; Lei LIU ; Shouqing LI ; Yan TAN
Chinese Journal of Immunology 2017;33(1):62-65
Objective:To observe the inhibitory effect of haploid vaccine pcDNA-CEA625-667 and three tandem repeats of minigene DNA vaccine pcDNA-triCEA625-667 derived from CEA gene on tumor in mice bearing tumor and the changes of survival time. Methods:The experimental animal model of mouse liver cell carcinoma was established and the mice were immunized with pcDNA-CEA625-667 and three series of DNA vaccine. Some of the mice were treated with normal saline as control group. The growth curve of tumor growth curve was recorded and the effect of vaccine on the survival time of tumor bearing mice was observed. Results:Compared with the normal saline control group,the two vaccines were able to significantly inhibit the tumor size and growth rate ( P<0. 01 ) of CEA positive tumor bearing mice,the inhibition of pcDNA-triCEA625-667 vaccine group was significantly better than the pcDNA-CEA625-667 vaccine group (P<0. 01),while the two were not inhibited tumor growth in CEA negative tumor bearing mice. The average survival time of the pcDNA-CEA625-667 vaccine group was(48. 50±6. 73)d,and there was significant difference (P<0. 01) compared with the saline control group ( 39. 00 ± 6. 64 ) d. The survival time ( 48. 50 ± 6. 73 ) d of the pcDNA-triCEA625-667 vaccine group was significantly higher than that of the normal saline control group and the pcDNA-CEA625-667 vaccine group (P<0. 01). The survival time of CEA negative tumor bearing mice could not be prolonged in the two groups. Conclusion:Either the haploid or the three series of the DNA vaccine,were able to significantly inhibit tumor growth rate (P<0. 01) and significantly prolong the survival time (P<0. 01) of CEA positive tumor bearing mice,but they had no therapeutic effect on CEA negative tumor bearing mice.