1.Proteomic identification of proteins in exosomes of patients with atherosclerosis
Mei JIANG ; Jing QUAN ; Heng ZHANG ; Qianqian DING ; Meng XIANG ; Dan MENG ; Ning SUN ; Sifeng CHEN
Chinese Journal of Pathophysiology 2016;32(8):1525-1526
AIM:Atherosclerosis primarily involved systemic arteries .Luminal surface , a monolayer of endothelial cells , of artery directly exposes to blood and is susceptible to active substances in the blood .Exosomes contain significantly amount of proteins and RNAs .Ex-osomes can be good and bad for cells , depending on their component .Thus, exosomes may contribute to atherosclerosis by affecting endothelial cells .This study analyzed the relationship of exosome proteins and atherosclerosis .METHODS: Fifty-six patients and healthy subjects were recruited and divided into two comparisons:healthy subjects vs atherosclerosis ( HS vs AS) , and hypertension vs hypertension plus atherosclerosis ( HT vs HT+AS) .Serum exosomes were decoded by protein mass spectrometry .The protein profile and function were analyzed by gene ontology ( GO) .RESULTS:It was found that five child terms repeatedly appeared in “response to stimulus” and “immune system process” of BP of the two categories ( HS vs AS and AS vs HT+AS):“positive regulation of innate immune response”,“immune response-activating signal transduction”,”activation of innate immune response”,“innate immune re-sponse-activating signal transduction” and “innate immune response activating cell surface receptor signaling pathway ”.Two child terms repeatedly showed in “binding” of MF of the two categories:“antigen binding” and “enzyme binding”.Two proteins, PSMA6 and PSMA7, were repeatedly shown in the two categories .CONCLUSION:GO analysis was utilized for structure hierarchy “tree” to illustrate these proteins involved in various terms in BP , CC and MF.The PPI analysis supplied proteins which may play potentially im-portant roles in AS process .Innate immune system and blood coagulation pathway contribute to AS formation .The proteins, PSMA6, PSMA7 and Annexin A2, may can be the new target proteins for prevention and treatment of AS .
2.Proteomic identification of differentially expressed proteins in blood exosomes of patients with hypertension
Jing QUAN ; Mei JIANG ; Heng ZHANG ; Qianqian DING ; Meng XIANG ; Dan MENG ; Ning SUN ; Sifeng CHEN
Chinese Journal of Pathophysiology 2016;32(8):1525-1525
AIM:To analyze the proteins included in exosomes derived from blood of patients with hypertension and seek the main pathologi -cal changes in hypertension .METHODS:Forty-seven patients and healthy subjects were recruited and divided into two comparisons :healthy subjects vs atherosclerosis ( HS vs AS) , and atherosclerosis vs hypertension plus atherosclerosis ( AS vs HT+AS) .We extrac-ted exosomes from blood and utilized LC-MS/MS to identify the protein expression .We used GO analysis to established the hierarchy programs of biological process and molecular function .PPI was used to find the proteins related to the terms .RESULTS:It was found that three final child terms repeatedly shown in BP of the two categories ( HS vs AS and AS vs HT+AS):“signal transduction in re-sponse to DNA damage”,“response to zinc ion”, and“platelet aggregation”.It was found that two final child terms in MF of the two categories:“interleukin 2 receptor binding” and“ploy(A) RNA binding”.The proteins, PSMA6, PSMA7 and CA2, were related to the terms in the two categories .CONCLUSION: We discovered that the exosome proteins may indicate the pathological changes in hypertension through the biological processes related with the specific proteins .These specific proteins, such as VCL, PSMA6, DP, AKAP, ATP5B and CA2, can be the new indicators for severity of hypertension and new therapeutic targets .
3.Epidermal growth factor receptor involvement in cardiovascular remodeling in renovascular hypertensive rats
Mei FENG ; Wenhao TENG ; Wenwen JIANG ; Jizhou XIANG ; Hui LIU ; Lei YANG
Chinese Pharmacological Bulletin 2016;32(5):625-630,631
Aim To determine the possibilities and mechanisms of EGFR, a receptor protein tyrosine kinase associated with many important cellular processes re-sponsible for cardiovascular remodeling in renovascular hypertensive rats. Methods 2K1C hypertensive rats were used in the present study. Blood pressure was measured with the tail-cuff method. LVMI and his-topathological changes in the cardiovascular system were analysed. EGFR expressions of aorta and myocar-dium as well as phosphorylation levels of ERK in hy-pertensive rats were detected by immunohistochemistry and Western blot analysis, respectively. Results Sys-tolic blood pressure was markedly increased 2 weeks after 2K1C surgery. Cardiovascular remodeling induced by hypertension was confirmed by elevated LVMI, pro-liferation of collagen fibers in myocardial interstitium, histopathological changes in cardiovascular system and increased IMT of thoracic aorta 6 weeks after 2 K1 C surgery. Compared with sham rats, EGFR expression in the ventricular myocardium of 2 K1 C rats was signifi-cantly increased at 6 weeks ( P<0. 05 ) , and the EG-FR/GAPDH ratio was higher in 2 K1 C rats with higher systolic blood pressure ( P < 0. 05 ) . Phosphorylation level of ERK 1/2 was upregulated correspondingly in 2 K1 C rats ( P <0. 01 ) . Increased EGFR expression was also found in aortas of 2K1C rats, particularly in tunica intima and media. Conclusion EGFR and its down-stream kinases ERK 1/2 are involved in cardio-vascular remodeling in association with the severity of hypertension in renovascular hypertensive rats.
4.A rat carotid atherosclerosis modal induced by artery clamping with high-cholesterol diet and vitamin D3
Xiang XU ; Mei WU ; Bin LI ; Xuesong JIANG ; Runze ZHOU ; Guangyi LIU ; Xudong PAN
International Journal of Cerebrovascular Diseases 2013;(4):288-292
Objective To investigate the feasibility of a rat carotid atherosclerosis model induced by artery clamping with high-cholesterol diet and vitamin D3.Methods Twenty Wista rats were randomly divided into either a general diet group (n =5) or a high cholesterol diet group (n =15).After clamping the right common carotid arteries of the rats in the high cholesterol diet group,they were fed with high fat diet,and vitamin D3 (600 000 IU/kg) was injected intraperitoneally.The lipid levels of the general diet group and the high cholesterol diet group were detected at 6 and 12 weeks respectively.The bilateral cormmon carotid arteries were selected for preparing paraffin sections and were stained with HE staining.The pathological changes in blood vessels were observed.Results The levels of serum total cholesterol at 12 weeks (2.803 ± 1.307 mmol/L vs.1.513 ±0.281 mmol/L; P=0.017) and low-density lipoprotein cholesterol (0.660± 0.260 mmol/L vs.0.311 ±0.078 mmol/L; P =0.003) in the high-cholesterol diet group were significantly higher than those at 6 weeks.The histopathological examination showed that the common carotid artery intimas on the clamping sides were incomplete,the foam cell deposition was observed under intima,the atherosclerotic plaques or fibrous plaques were observed on the surface of cavity,inside the plaques contained necrotic tissue,and thrombosis was observed in the cavity.The common carotid artery intima-media thickness in the general diet group (n =5) at 12 weeks was 8.3 ± 1.1 μm.The sham-operated sides (n =20) and clamping sides (n =20) were 8.8 ± 0.7 μm and 97.4 ±25.7 μm,respectively.There were significant differences among the three groups (F =116.313,P=0.000).The clamping sides in the high-cholesterol diet group were significantly higher than the shamoperated sides in the high-cholesterol diet group (P=0.000) and the general diet group (P =0.000).Conclusions Common carotid artery clamping with high-cholesterol diet and vitamin D3 is a simple and feasible method for inducing a rat carotid atherosclerosis model.
5.Correlation between chest-abdomen longitudinal diameter ratio,total lung area,lung longitudinal diameter and fetal lung development
Fan JIANG ; Na CHEN ; Mei PENG ; Xiang XIE ; Pin LUO ; Xinshu ZHANG
Chinese Journal of Ultrasonography 2011;20(10):864-866
ObjectiveTo evaluate the application of chest-abdomen longitudinal diameter ratio,total lung area and lung longitudinal diameter in different gestational ages by two-dimensional ultrasonography.MethodsFive hundred and ninty-two fetus of normal singleton pregnancy between 18 and 40 weeks were ultrasonic scanned by standard 4-chamber view and fetus torso coronal section.The heart area,thoracic area,bilateral lungs pointed to the corresponding diaphragmatic dome gallery on the top of the vertical distance,diaphragmatic dome iterated top to the bottom of the fetal bladder distance were measured respectively.Related regression analysis was done on fitting total lung area and longitudinal diameter with pregnant increasing.ResultsA normal pregnancy fetal total lung area and lung longitudinal diameter were increased as long as the gestational age.The best-fit regression equations of total lung area was:Y =0.83 X +13.894,R2 =0.914;Lung longitudinal diameter was:Y =0.669 X + 3.124,R2 =0.892.Chest-abdomen longitudinal diameter ratio from 18 to 40 weeks gestation was in constant range:0.44 - 0.59.Conclusions Chest-abdomen longitudinal diameter ratio from 18 to 40 weeks gestation is in constant range.The normal pregnancy fetal total lung area and lung longitudinal diameter were increased as long as the gestational age.These parameters may be useful for the prenatal assessment of lung development.
6.Field epidemiological study on news reports that related to public health emergencies
Shun-Xiang ZHANG ; Xue-Mei LI ; Nian-Ci LUO ; Shu-Jiang MEI ; Li-Juan JIANG
Chinese Journal of Epidemiology 2013;34(12):1203-1207
Objective All news reports (NR) that were related to public health emergency (PHE) were collected from the Southern Metropolis Daily (SMD) to explore the characteristics of epidemiology in the fields.Methods Based on the theory of communication that including both case and text analysis,qualitative analysis on all the NR regarding PHE published in SMD from the years of 2008 to 2012,was carried out and input to database using the EpiData.Numbers of articles as indicators were compared to show the yearly change of different types of events.Various features of the NR including coverage,source of information,location of the incident,style and size of news,with or without editorials etc.were statistically analyzed by SPSS version 18.0.Results Among all the 998 reports related to PHE,higher proportion was found in the events of Infectious diseases (35.3%)and food safety (34.1%) respectively.Events on vaccines and drugs used for disease prevention and control (8.9%),environmental pollution caused incidents (8.0%) appeared to be less frequent.Events related to occupational disease,poisoning,bioterrorism and biochemical events were rare.Looking at the monthly distribution of reports,we noticed that the peaks occurred in 2008 and in 2009,which were caused by the Melamine-contamination events and the 2009 H1N1 pandemic.Between 2010 and 2012,figures of monthly reports were smooth,including some critical events from the interests of the media.Most events took place in Guangdong province (34.3%) and other provinces (50.9%),with some were from Hong Kong,Macao and Taiwan regions (9.5%).However,international events (5.2%) were less seen.Extensive coverage accounted for 17.6% of all of reports,and 11.5% allotted the editorials or other forms of in-depth reports.Most of the source of reports on infectious diseases and food safety were from the official release,however.The main sources of occupational diseases and poisoning,vaccines and drug incidents,environmental pollution related incidents were reported by active journalists through interview.Reports on hand,foot and mouth disease,influenza,milk safety,AIDS and lead pollution showed continued concern in the past five years by SMD.Conclusion NR on public health emergencies by SM had encompassed all 10 categories-related events formulated by the Ministry of Health.Sustained and in-depth coverage were more commonly seen.Fieldepidemiologists should learn interdisciplinary sciences on the theory and methodology of communication.They also need to interact with media people during the whole processes of public health emergency preparedness and responses.
7.Effects of electroacupuncture versus sibutramine on adipocyte products in obesity rats
Yujin FAN ; Bin XU ; Xiaoren XIANG ; Jinhong YUAN ; Zhicheng LIU ; Yong SUN ; Xinjun WANG ; Mei CHEN ; Yani LU ; Junzuo JIANG
Chinese Journal of Tissue Engineering Research 2008;12(11):2189-2192
BACKGROUND:Both acupuncture and moxibustion and drug ale effective methods to treat simple obesity.Whether electroacupuncture is different from sibutramine to affect adipocyte products in obese organisms?OBJECTIVE:To observe the different effects of electroacupuncture and sibutramine on adipocyte produces in obesity rats and to investigate the mechanism of weight reduction by acupuncture and moxibustion.DESIGN:Layering-randomized controlled study.SETTING:Laboratory of Acupuncture and Pharmacology.Nanjing University of Chinese Medicine.MATERIALS:12045-day-old male SD rats were provided by Shanghai Experimental Animal Center.The experiment was in accordance with animal ethics standards.Sibutramine hyrdrochloride capsule was made in Fuling Pharmaceutical Factory of Taiji Group(batch number:X20010279).Electro-acupuncture apparatus(LH402A acupoint neutoelectrical stimulation equipment)was made in Beijing Huawei Chanye Developing Company.METHODS:This study was performed at the Laboratory of Acupuncture and Pharmacology,and Expefimental Animal Center,Nanjing University of Chinese Medicine from October 2004 to November 2006.①100 male SD rats underwent high-fat diet.Fifty-four alimentary obesity rats were randomly divided into electroacupuncture group,sibutramine group and obesity group.wim 18 rats in each group.Another 18 rats were regarded as a normal group.②Rats in the electroacupuncture group were given electrical stimulation(2/15 Hz,4 mA)at lateral Housanli and Neiting acupoints,contralateral alternation every other day.Rats were fixed with self-made binding instrument,and rats maintained a consciousness but not an intense struggle.The electricai stimulation lasted for 15 minutes every day,one day was an interval every 6 days.and the electroacupuncture was given for 49 days and 42 times in total.Rats in the sibutramine group were ingragastrically administrated with 0.2 g/L sibutramine hydrochloride(2.0 mg/kg/d).Rats in the control group and obesity groups were fixed the same as electroacupuncture group but not given electrical stimulation.MAIN OUTCoME MEASURES:①Obese marker:To detect body mass,body length,weight reduction rate and body mass/body fat mass,and to measure total cholesterol(TC),high density lipoprotein cholesterol(HDL-C)and triacylglycerol (TG)with biochemical chromatometry;②insulin sensitivity index:To measure fasting blood glucose,insulin and insulin sensitivity index with enzyme-linked immunosorbent assay;③adipocyte secreting regulatory substance level:To measure serum resistin,leptin,adiponectin,tumor necrosis factor-α(TNF-α),interleukin-6(IL-6)and its soluble receptor level with enzyme-linked immunosorbent assay.RESULTS:Among 120 rats,54 alimentary obesity rat models and 18 normal rats were included in the final analysis.①Obesemarker:Bodymass,bodyfatmass,cholesterol and TG levels in the obesity group were significantly higher than in the normal group(P<0.01).Body mass,body fat mass,body mass/body fat mass,cholesterol and TG levels in the electroacupuncture group were significantly lower than in the obesity group(P<0.01).TG level in the sibutramine group was significantly lower than in the obesity group(P<0.01).Weight reduction rate in the electroacupuncture group was significantly higher than in the sibutramine group(P<0.01).Body mass/body fat mass in the electroacupuncture group was significantly lower than in the sibutramine group(P<0.01).②Insulin sensitivity:Blood glucose and serum insulin levels in the obesity group were significantly higher than in the normal group,and insulin sensitivity index was significantly lower than in the normal group(P<0.01).Blood glucose and insulin levels in the electroacupuncture and sibutramine groups were significantly lower than in the obesity group,but insulin sensitivity index was significantly higher than in the obesity group(P<0.01).③Adipocyte secreting regulatory substance level:Adiponectin in the electroacupuncture group was significantly higher than in the obesity group (P<0.01).Resistin and TNF-αin the electroacupuncture group was significantly lower than in the obesity grollp(P<0.01).Resistin level in the sibutramine group was significantly lower than in the obesity group(P<0.01).CONCLUSION:The effect of electroacupuncture on abnormally secretary products of adipose tissues in obesity rats is superior to the effect of sibutramine.This may be one of important mechanisms of weight reduction by acupuncture and moxibustion,and prevention and cure of obesity and its related diseases.
8.The effect of injection of bone mesenchymal stem cell into the joints of collagen-induced arthritis rats
Xi-lan YANG ; Peng ZHANG ; Mei-jiang FENG ; Hui-wei HE ; Chen QU ; Jing-jing HUANG ; Xiang LU
Chinese Journal of Rheumatology 2011;15(11):736-738
ObjectiveTo evaluate the effect of Bone mesenchymal stem cells(BMSCs) intraarticular injection on the pathological development of collagen-induced arthritis(CIA) rats.MethodsCIA Wistar rats were divided into 2 groups:the saline injection control group and the BMSCs injection group.BMSCs were isolated from normal rats and cultured in vitro till P2 generation,which were injected into the affected the ankle joint consequently.After 2,4,8,12 weeks of processing,the following indexes were observed:arthritis score and X-ray.Twelve weeks later,all of the rats were sacrificed and the following histological indexes were evaluated:cartilage destruction and cartilage extracellular matrix secretion.Paired samples t-test was used for the statistical comparison.ResultsAll the arthritis indexes of the BMSCs group were much lower.than those of the saline control group.The arthritis scores were 3.18±0.62 vs 3.84±0.35 (at week 2),3.45±0.28 vs 5.96±0.48(at week 4),3.86±0.23 vs 6.75±0.36(at week 8),4.23±0.43 vs 7.86±0.66 (at week 12) respectively.X-ray scores were 2.04±0.21 vs 2.72±0.15(at week 2),2.52±0.47 vs 4.06±0.38 (atweek 4),3.56±0.29 vs 4.35±0.36 (at week 8),3.73±0.43 vs 4.86±0.62(at weekl2) respectively and the histological evaluation indexes were 2.34±0.22 vs 3.52±0.55 (at week 12).ConclusionBMSCs injection may inhibit the pathological development of CIA rats,and it may be a promising approach for the treatment of rheumatoid arthritis.
9.A study on the level of N -terminal pro -B -type natriuretic peptide in evaluating the cardiac function of pregnant women with cardiac disease
Fang ZHOU ; Li-Ping CHEN ; Mei-Fen WENG ; Jin-Xiang JIANG
Journal of Preventive Medicine 2015;(4):351-354
Objective To explore the relationship between the cardiac function of pregnant women with cardiac disease and the level of N -terminal pro -B -type natriuretic peptide (NT -pro -BNP).Methods A total of 74 pregnant women with cardiac disease and 80 healthy pregnant women were enrolled in the study and the NT -pro -BNP was detected and compared.The concentrations of NT -pro -BNP in different trimester were compared among the healthy pregnant women, and the correlation between the cardiac function of pregnant women with cardiac disease and the level of NT -pro -BNP were analyzed.Results The median of concentrations of plasma NT -pro -BNP in pregnant women with cardiac disease was 352.50 pg/mL,which was significantly higher than that in healthy pregnant women 58.00 pg/mL(P <0.01 ).The cardiac function was positively correlated with the level of NT -pro -BNP (P <0.01 ).The concentration of plasma NT -pro -BNP during the late trimester(49.00 pg/mL)was significantly higher than that of the early trimester(49.00 pg/mL) among the healthy pregnant women (P <0.01 ).Conclusion It is of great clinical value for the concentration of plasma NT-pro -BNP in evaluating the cardiac function of pregnant women with cardiac disease,and the pregnancy may influence the cardiac function.
10.Differentially expressed genes associated with apoptosis in human rhabdomyosarcoma cells infected by enterovirus 71
Xiang LI ; Xueling HOU ; Hongjun PENG ; Mei SHI ; Qingbo JIANG ; Xiping LIU ; Yuhua YAO ; Caizhen HE ; Weifeng SHI
Chinese Journal of Microbiology and Immunology 2012;32(8):718-724
Objective To explore the different apoptotic gene expressions and apoptotic signaling transduction of human rhabdomyosarcoma (RD) cells infected by enterovirus 71 (EV71) in different stage.Methods The survival of EV71-infected RD cells was observed by trypan blue staining.The apoptotic morphology and rates of RD cells were surveyed and detected by Annexin V-FITC/PI staining and flow cytometry,respectively.PCR array was employed to analyze 88 apoptotic gene expressions from EV71-infected RD cells at 8 h and 20 h postinfection (p.i),respectively.Results After RD cells were infected with EV71 (MOI =5) at 8 h p.i,the viability was significantly decreased.Flow cytometry data demonstrated that the apoptotic rates of EV71-infected RD cells had increased to 18.0% and 19.1% at 20 h p.i in early and later stage respectively.RT-PCR array studies revealed significant variations in the expression of apoptotic genes.Among 88 genes,only the expression of IFN-β1 was upregulated 5.22 folds,whereas 47 genes including ACIN1,Akt,Apaf1,caspase and CIDEB were found to be downregulated that were lower than 2 folds at 8 h p.i.However,28 genes including FasL,CD40L,TNF,caspase-10 and caspase-3 were induced more than 2 folds after EV71 infection at 20 h.Conclusion The downregulation of apoptosis-related genes is associated with viral apoptosis-suppressing effect in RD cells in the early stage of EV71 infection.The death receptor signaling pathways including Fas/FasL and TNF/TNFR are activated to induce cell apoptosis in the late stage of EV71 infection.Moreover,host cell can also inhibit apoptosis by regulating signal pathway of CD40/CD40L,NF-κB/RelA and PI3K/Akt activation.