1.Mechanism of the effect of rAd-APN gene on atherosclerosis in ApoE-/-mice
Xuemei WANG ; Qin WEI ; Chun ZHANG ; Tao JIANG ; Mingjun DUAN ; Yining YANG
Acta Laboratorium Animalis Scientia Sinica 2016;24(2):175-183
Objective Atherosclerosis ( AS) is a common pathological basis of cardiovascular diseases.Adiponec-tin ( APN) has been shown to have an anti-AS effect, and the underlying mechanisms, however, are largely unknown.Nu-clear transcription factorκB ( NF-κB) has also been regarded as a proatherogenic factor, mainly because of its regulation of a variety of the proinflammatory genes linked to AS.It is hypothesized that the inhibitory effects of APN on AS is through the inhibition of NF-κB signaling pathway.The aim of this study was to test the hypothesis via investigation and validation of the inhibitory effect of APN on AS in ApoE-/-mice, and to delineate the roles of NF-κB signaling pathway in modulating the APN effect on AS in vivo.Methods APN overexpression in ApoE-/-mice were mediated by transfecting adenovirus bearing a vector encoding for APN and enhanced green fluorescent protein ( Ad-APN-eGFP) .The AS in ApoE-/-mice was induced by feeding a high-fat diet.To validate the inhibitory effect of the adenovirus mediated APN overexpression on AS in the ApoE-/-mice.120 male ApoE-/-mice aged 12 weeks were randomly and evenly assigned into two groups (60 mice per group), and were fed with a high-fat diet to induce AS.At 0 day, 2, 4, and 6 weeks of high-fat diet feeding.The 2 groups of mice were injected intravenously in the tail with either 100 μL (3.0 ×108 p.f.u) of Ad-eGFP virus ( control group) or the same amount of Ad-APN-eGFP virus ( APN group) .Blood samples and aortic tissues were taken at 0 day, 4, and 8 weeks of high-fat diet feeding.For the blood samples, FABA was used to analyze the concentrations of blood lipids and ELIZA was used to test the concentrations of serum APN.For the aortic tissues, oil red O staining was used to detect the surface lesion percentage.Masson staining was used to evaluate the collagen content and fibrous cap thickness of the plaque area.Immunofluorescence method was used to detect APN and NF-κB p65 expression.Western blot was used to de-tect the expressions of APN,nuclear NF-κB p65 and the downstream factors of NF-κB pathway.Results APN inhibited the formation of atherosclerotic plaque in ApoE-/-mice.The lesion formation in aortic sinus was significantly inhibited ( P<0.01).Compared with the control group, the oil red O staining showed that the surface area ratio of atherosclerotic le-sions was decreased significantly in the Ad-APN group ( P<0.001 ): the percentage of surface lesions in the 4 weeks groups was 27.78 ±8.64 vs.33.02 ±5.18 (%);the 8 weeks groups was 31.58 ±5.87 vs.52.16 ±5.79 (%) .As the serum APN was increased,the concentration of TC, TG and LDL-C were significantly decreased( P<0.001 for all) , and the growth of body weight was slowed down(P<0.05).APN effectively inhibited the expression of NF-κB nuclear protein p65 and inflammatory factors.Conclusions Adiponectin reduces the inflammatory reactions in atherosclerosis through in-hibiting the NF-κB pathway.
2.Effect of spironolactone on cardiac remodeling after acute myocardial infarction
Chun-Tao WU ; Zhong-Hua WANG ; Zhu-Qin LI ; Lan-Feng WANG
World Journal of Emergency Medicine 2013;4(1):48-53
BACKGROUND: Few studies have reported the effect of aldosterone receptor antagonist (ARA) on myocardial remodeling after acute myocardial infarction (AMI). This study was undertaken to investigate the preventive effect of ARA on myocardial remodeling after AMI.METHODS: A total of 616 patients who had been admitted into the CCU of the First Affiliated Hospital of Harbin Medical University from January 2008 to January 2010 were studied prospectively. Only 528 patients were observed completely, including 266 of the control group and 262 of the treatment group. There was no statistical difference in age, gender, medical history, admission situation, and treatment between the two groups (P>0.05). The preventive effects of spironolactone on cardiac remodeling, left ventricular function, renal function and blood levels of potassium were evaluated by echocardiography, serum potassium and serum creatinine at one-month and one-year follow-up.RESULTS: The echocardiography indicators such as LVESD, LVEDD, LVEF, LAD-ML and LAD-SI were signifi cantly improved in the treatment group compared with the control group at one year (P<0.05). In the treatment group, LVESD, LVEDD, LVPWT, LVEF, LAD-ML and LAD-SI were more significantly improved at one year than one month (P<0.05,P=0.007 to LVEF), and in the control group LVEF was more signifi cantly improved at one year than one month (P=0.0277). There were no signifi cant differences in serum potassium and serum creatinine levels between the two groups.CONCLUSION: On the basis of conventional treatment, the early combination of low-dose spironolactone (20 mg/d) could inhibit cardiac remodeling at late stage and prevent heart failure.
3.Effect of andrographolide on the mRNA expression of MexAB-OprM efflux pump of Pseudomonas aeruginosa PAOI strain
Hong-Tao LI ; Chun-Ming WU ; Hui-Min QIN ; Guo-Jun LI ; Jian-Xin SONG ;
Chinese Journal of Infectious Diseases 2001;0(06):-
Objective To develop a real-time polymerase chain reaction(PCR)system to determine transcriptional level of MexAB-OprM multidrug efflux pump gene and to investigate the impact of androgra- pholide on MexAB-OprM gene transcription in Pseudomonas aeruginosa.Methods The fragments of mexB gene of mexAB-oprM operon and 30S rRNA gene rpsL were amplified and cloned into two plas- mids respectively.These plasmids were used as external standards for real-time PCR.Real-time PCR was applied to measure the mRNA transcripition of mexB and rpsL gene in Pseudomonas aeruginosa growing in medium with different concentrations of andrographolide.Results The plasmids for standard curve were constructed successfully.The relative mexB mRNA expressions in 50,100,150 and 200?g/mL andrographolide were 0.04?0.03,0.06?0.07,0.09?0.03 and 0.04?0.03 respectively, which were significantly lower than that in the control(0.24?0.04,P0.05).Conclusion Andrographolide can reduce the transcriptional level of MexAB-OprM,which may he one mechanism for its anti-infection effect.
4.Histological changes of the right ventricular remodeling in Xinjiang Marmota baibacina adapted to high-altitude hypoxia
Mei MA ; Tao JIANG ; Chun ZHANG ; Qin WEI ; Minjun DUAN ; Xuemei WANG ; Zhiqiang LI ; Xi SHOU ; Qilong CHEN
Acta Laboratorium Animalis Scientia Sinica 2014;(2):45-47
Objective To investigate the histological changes of the right ventricular remodeling in Xinjiang Mar-mota baibacina which are adapted to high-altitude hypoxia .Methods Hematoxylin and eosin and Masson staining was used to examine the pathological changes and degree of fibrosis , and immunohistochemical technique was used to detect connexin 43 ( CX43) expression in the right ventricular myocardium .Results In the right ventricular myocardium , cardio-myocyte hypertrophy and increased amount of collagen fibers were observed .The right ventricular hypertrophy index and weight index of Marmota baibacina were significantly higher , and expression of CX43 was significantly reduced than that of normal rats ( P<0.05 for both ) .Conclusions High-altitude hypoxia induces right ventricular remodeling in Xinjiang Marmota baibacina.It can be used as an ideal animal model for studies on the mechanisms of hypoxia adaptation on high -altitude plateau .
5.Study about proliferation and apoptosis of megakaryocytes in patients with myelodysplastic syndromes.
Xiao LI ; Quan PU ; Yi-Zhi LIU ; Jun SHI ; Ying TAO ; Chun-Kang CHANG ; Qin-Yan JIANG ; Wei HUANG
Journal of Experimental Hematology 2002;10(1):40-43
In order to observe the proliferative and apoptotic situation of megakaryocytes in patients with myelodysplastic syndromes (MDS). CD41 immunoenzyme labeling (alkaline phosphatase anti-alkaline phosphatase APAAP)/DNA in situ end labelling (ISEL) double stained techniques was used onto plastic cold embedded bone marrow sections in 29 MDS patients to analyse the proliferative and apoptostic characterization of megakaryocytic line with 14 cases of iron deficient diseases (IDA) as control. The results showed that the mean CD41 positive cell number in MDS group was (26.23 +/- 8.18) /mm(2) with a count of (15.64 +/- 7.11) /mm(2) in control group (p < 0.05). The small-micro megakaryocytes in MDS is much higher than that in IDA group (P<0.01). There was a positive co-relation between total megakaryocytes and small-micro megakaryocytes count in MDS (r = 0.702, p<0.01). Some megakaryocytes distributed abnormally around trabecula and formed small or large clusters. Apoptotic megakaryocytes in MDS occupied 4.40% and 9.32% of all CD14 positive cells and all apoptotic cells respectively (p > 0.5 comparing with control). Apoptosis in megakargocytic line occurred only in small-micro megakaryocytes and showed positive co-relation to the number of micro-megakaryocytes. Some apoptotic cell with morphologic characters of megakaryocytes expressed no CD41. It is concluded that overproliferation of megakaryocytes exists in MDS. Apoptosis occurring in micro-megakaryocytes may be a kind of physiological response to abnormal megakaryopoicsis in MDS. No obvious increased apoptosis of megakaryocytes in MDS was found perhaps due to lack of surface antigens CD41 in some later stages of apoptotic cell.
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6.Effect of andrographolide on QS regulating virulence factors production in Pseudomonas aeruginosa.
Hong-tao LI ; Hui-min QIN ; Wei-hua WANG ; Guo-jun LI ; Chun-ming WU ; Jian-xin SONG
China Journal of Chinese Materia Medica 2006;31(12):1015-1017
OBJECTIVETo investigate the effect of andrographolide on virulence factors production in Pseudomonas aeruginosa.
METHODGrowth rate, pyocyanin, proteolytic activity and elastase activity were measured with or without the presence of andrographolide. The effect of andrographolide on pyocyanin production, proteolytic activity and elastase activity in PAO-JP2 was investigated simultaneously.
RESULTThe andrographolide did not affect the growth of PAO1 in planktonic culture. The production of pyocyanin, proteolytic activity and elastase activity were significanthy suppressed in P. aeruginosa cultures grown in the presence of andrographolide. However, these effects were not observed in PAO-JP2.
CONCLUSIONThe inhibiting effect of andrographolide on virulence factors production in P. aeruginosa may play a role in its anti-infection activity.
Andrographis ; chemistry ; Anti-Bacterial Agents ; pharmacology ; Diterpenes ; isolation & purification ; pharmacology ; Pancreatic Elastase ; metabolism ; Peptide Hydrolases ; metabolism ; Plants, Medicinal ; chemistry ; Pseudomonas aeruginosa ; growth & development ; metabolism ; pathogenicity ; Pyocyanine ; metabolism ; Virulence Factors ; metabolism
7.The value of combined detection of LP-PLA2 and sd-LDL in the auxiliary diagnosis of atherosclerosis
Mingming QIN ; Yuxin HUANG ; Xuemei CHEN ; Yuxuan WANG ; Qingsong TAO ; Chun PU
Chinese Journal of Laboratory Medicine 2019;42(1):38-43
Objective To investigate the auxiliary diagnostic value of combined detection serum lipoprotein-associated phospholipase A2 (LP-PLA2) and small and dense low-density lipoprotein (sd-LDL) in atherosclerosis.Methods The subjects were divided into experimental group and control group by random block design from May 2017 to January 2018,in the First Affiliated Yijishan Hospital of Anhui Wannan Medical College.The experimental group selected 125 AS patients with clinical diagnosis and confirmed by angiography,and 55 healthy subjects were chosen as the normal control group at the same time.Serum samples were collected within 24 hours after admission,and the level of LP-PLA2,sdLDL-C,low density lipoprotein cholesterol (LDL-C),triglyceride (TG),total cholesterol (TC) and high-sensitive C reactive protein (hs-CRP) were unified detection.The t test,single factor variance analysis and Mann-Whitney U and multivariate logistic regression analysis were used to analyze the data.Results LP-PLA2,sd-LDL,LDL,TC,TG and hs-CRP of the AS group were all higher than those of the healthy control group (Z=5.279,6.663,6.012,5.863,5.508 and 2.845,respectively,P<0.05).Logistic regression analysis showed that serum LP-PLA2,sd-LDL and hs-CRP level was an independent risk factor for predicting atherosclerosis (OR=1.008,P=0.003;OR=8.282,P=0.012;OR=1.158,P=0.009).The sensitivity of LP-PLA2,sd-LDL,LDL-C,TC,TG,hs-CRP to AS was detected separately (57.6%,73.6%,85.6%,83.2%,76.8%,80.0%),and the specificity was (89.1%,78.2%,67.3%,69.1%,74.5%,52.7%).The ROC curve showed that the diagnostic efficacy of LP-PLA2 and sd-LDL combined detection was 0.854,higher than sd-LDL,LDL-C,TC,TG,LP-PLA2 and hs-CRP (0.811,0.782,0.775,0.758,0.747 and 0.633,respectively).In addition,the levels of both increased with the aggravation of arteriosclerotic lesion(x2=7.954,P=0.019;x2=11.44,P=0.003).The levels of LP-PLA2 and sd-LDL in patients with AS were not significantly different between different lesions (x2=8.042,P=0.09;x2=5.952,P=0.203).There was no significant difference between serum LP-PLA2 and sd-LDL level and sex,age,smoking,hypertension and diabetes (Z1=0.398,0.719,0.619,0.098 and 1.338 respectively,Z2=0.942,0.027,0.894,0.375,0.783,respectively,both P1 and P2 were>0.05).Conclusions sd-LDL combined with LP-PLA2 has high sensitivity and specificity in the prediction of AS,which makes up for the deficiency of individual detection;sd-LDL and LP-PLA2 serum level has nothing to do with the lesion and has a positive correlation with the degree of lesions.It is not easily affected by other risk factors and can be used as a risk factor for predicting the occurrence of AS.
8.Effect of profound hypothermia on cytochrome C release and cell apoptosis in the hippocampus after global ischemia in rats
Hua-Ping QIN ; Chang-Chun YANG ; Yi ZHANG ; Tao-Yin XIE ; Qiang WANG ; Feng ZHI ; Ji-Yao JIANG
Chinese Journal of Neuromedicine 2012;11(2):138-140
Objective To detect the effect and role of profound hypothermia on cytochrome C release and cell apoptosis in the hippocampus after global ischemia in rats. Methods We established the animal model of cardiopulmonary bypass (CPB) in 24 rats which were randomized into 3 even groups:blank control group,normothermia ischemia group and hypothermia ischemia group.After the hippocampus samples were harvested from the 3 groups,Western blot was used to detect the expression of cytochrome C (Cyt-C) and TUNEL was used to measure the cell apoptosis. Results The expression of Cyt-C in the normothermia ischemia group increased significantly compared with the blank control group but decreased significantly compared with the hypothermia ischemia group (P<0.05).The rate of cell apoptosis in the normothermia ischemia group increased significantly compared with the blank control group but decreased significantly compared with the hypothermia ischemia group (P<0.05).Conclusions Profound hypothermia can inhibit cytochrome C release into the cytoplasm and thus reduce cell apoptosis in the hippocampus after global ischemia in rats. This may be one important mechanism underlying the protective effect of profound hypothermia.
9.Identification of human coronary artery endothelial cells obtained by coronary endovascular biopsy.
Shi-Yong YU ; Lan HUANG ; Yao-Ming SONG ; Ai-Min LI ; Jun QIN ; Xue-Jun YU ; Zhao-Hua GENG ; Gang ZHAO ; Chun-Mei LIN ; Chun-Rong TAO
Chinese Journal of Cardiology 2008;36(3):240-242
OBJECTIVETo develop a method to obtain and identify human coronary artery endothelial cells obtained during percutaneous coronary interventions (PCI).
METHODSCoronary guide wires were used to obtain endothelial cells from coronary arteries in 28 patients undergoing PCI. The cells were eluted from the wire tips and then purified by magnetic beads coated with anti-CD146 antibody. von Willebrand factor (vWF) was used as an immunocytochemical marker for endothelial cells. The cellular viability was evaluated by observing cell membrane integrity and energy-dependent uptake of DiI-labeled acetylated low-density lipoprotein.
RESULTSAn average of 96 coronary artery endothelial cells with good viability per patient were obtained by one guide wire. vWF identification showed their endothelial morphology and immunoreactivity.
CONCLUSIONThe viable coronary endothelial cells could be obtained during routine percutaneous coronary interventions combined with magnetic beads isolation technique. These cells may be used for further cellular functional analyses (such as immunocytochemistry and molecular biology) and expand our understanding on mechanisms of coronary artery diseases.
Biopsy ; methods ; Coronary Vessels ; cytology ; pathology ; Endothelium, Vascular ; cytology ; pathology ; Female ; Humans ; Male ; Middle Aged
10.Effect of allogenic adipose-derived stem cell grafting on serum biochemical indicators and sporting ability of endurance trained rats.
Hui-ying YANG ; Hui TAO ; Mei-chun YU ; Chun YANG ; Rong-mei QU ; Wen-qin ZENG ; Fu-hua DUAN ; Lei YU ; Jing-xing DAI ; Lin YUAN
Journal of Southern Medical University 2011;31(7):1159-1163
OBJECTIVETo observe the effect of transplantation of allogeneic adipose-derived stem cells (ADSCs) on serum biochemical indicators and sporting ability in highly endurance-trained rats from a fasciological perspective.
METHODSThe ADSCs were cultured in vitro. Forty male Wistar rats were randomized into 4 equal groups, namely the blank control group, overtraining (model) group, transplantation without training group and overtraining plus transplantation group. The rats in the two overtraining groups were subjected to exhaustive swimming for 1 week, and in the two transplantation groups, cultured allogeneic ADSCs (2×10(6)/ml) were injected via the tail vein. The exhaustion time in swimming and the serum levels of BUN, LDH, BLa, and Hb of the rats were recorded after the treatments.
RESULTSThe rats in the model group showed significantly increased serum BUN, LDH and BLa levels and decreased Hb level with a extended exhaustion time as compared with those in the blank control group (P<0.01). The BUN, LDH and BLa was significantly lower, Hb level higher and the exhaustion time significantly longer in the overtraining plus transplantation group than those in the model group (P<0.01).
CONCLUSIONSADSCs can effectively prolong the exhaustion time of rats during exhaustive swimming and enhance their sporting ability.
Adipose Tissue ; cytology ; Animals ; Blood Urea Nitrogen ; Fatigue ; prevention & control ; Hydro-Lyases ; blood ; Lactic Acid ; blood ; Male ; Physical Conditioning, Animal ; physiology ; Physical Exertion ; physiology ; Rats ; Rats, Wistar ; Stem Cell Transplantation ; methods ; Swimming