1.How does autophagy activation affect the apoptosis, proliferation and cycle of endothelial progenitor cells in rats?
Hui LIU ; Xiaoqiang LI ; Renda ZHU ; Qingyou MENG ; Huijun LU
Chinese Journal of Tissue Engineering Research 2015;(1):67-71
BACKGROUND:Previous studies have reported that rapamycin can affect the proliferation, migration and adhesion abilities of endothelial progenitor cels, but there is no report on the effect of autophagy, as wel as the interaction between autophagy and apoptosis. OBJECTIVE: To observe the effect of rapamycin activated autophagy activation on the proliferation, apoptosis, and cycle of endothelial progenitor cels. METHODS:Density gradient centrifugation was used to obtain mononuclear cels from bone marrow, and the mononuclear cels were inoculated on human fibronectin-coated culture plate.Then after cultured for 7 days the adherent cels colected were the endothelial progenitor cels. Different concentrations of rapamycin (0.01, 0.1, 1 and 10 μg/L) were added and cultured for 24 hours. Western blot was used to detect the LC3-II protein expression and monitor the induction of autophagy, flow cytometry was used to observe the cel cycle progression and apoptosis changes, and methylthiazolyldiphenyl-tetrazolium bromide colorimetric assay was used to observe the proliferation ability. Meanwhile, the ultrastructural changes were observed under transmission electron microscope. RESULTS AND CONCLUSION:Compared with the control group, there was no significant increasing of LC3-II protein expression of endothelial progenitor cels in 0.01 μg/L rapamycin group, and the LC3-II protein expression was in the high level. The LC3-IIprotein expression in the 1 μg/L and 10 μg/L rapamycin groups was higher than that in the control group, but lower than that in the 0.01 μg/L rapamycin group, which indicated that autophagywas particularly active when the concentration of rapamycin was 0.01 μg/L. The apoptosis of endothelial progenitor cels was increased with the increasing of concentration of rapamycin, and the proliferation rate was decreased with the increasing of concentration of rapamycin. The results indicate that activation of autophagy by bapamycin can promote the cel apoptosis, change the cel cycle significantly, and can inhibit the proliferation of endothelial progenitor cels.
2.Comparative study on clinical efficacy of different surgical methods for axillary osmidrosis
Likang ZHANG ; Bin MENG ; Liyu WANG ; Disheng MENG ; Jun LU ; Huagui FAN ; Jiahui FAN ; Lubirou GAO ; Jiangfeng ZHU
Journal of Regional Anatomy and Operative Surgery 2014;(2):172-174
Objective To investigate the most appropriate surgical methods,different surgical modes of osmidrosis and their clinical effi-cacy were observed. Methods Clinical data was collected from 200 cases of axillary osmidrosis from January 2011 to July 2013. These cases were divided into four groups of the traditional group,minimally invasive group,RF pen-frequency electric cautery group and improved curet-tage group. Results The traditional group(80 cases) has an average healing period of 18. 7 days for operative incision including 78 cured cases(97. 5%)and 2 significant improved cases(2. 5%). The minimally invasive group(60 cases)has an average healing time of 8. 6 days, among which there are 3 cured cases(5%),8 significant improved cases(13. 3%),16 improved cases(26. 7%),4 cases(6. 7%)with weak curative effect and 29 failed cases(48. 3%). In the RF pen-frequency electric cautery group(30 cases),there are 5 significant improved ca-ses(16.7%),8improvedcases(26.7%)and17failedcases(56.6%).Theimprovedcurettagegroup(30cases)withanaverageincision healingtimeof9.8dayscontains28curedcases(93.3%)and2significantimprovedcases(6.7%). Conclusion Thetraditionalgroup shows the best curative effect,nevertheless the incision needs a considerably amount of time to recover. The patients under the treatment of minimally invasive surgery or RF pen-frequency electric cautery can recover in short time but recrudescence always occur. The improved cu-rettage method,which is effective and safe,combines the advantages of traditional surgery and minimally invasive surgery. However,large scar left from this method still remains as its major disadvantage but the overall curative effect is satisfactory. The improved curettage is proved to be the most appropriate method for axillary osmidrosis.
3.Design and synthesis of photoaffinity biotin labelled 2'-O-propargyl-guanosine.
Lu-xin NA ; Xin LIU ; Zhuo-ming MENG ; Zhu GUAN ; Li-he ZHANG ; Zhen-jun YANG
Acta Pharmaceutica Sinica 2015;50(1):59-63
Photoaffinity labeling is widely applied to demonstrate targets of small molecule ligands. In this paper, biotin photoaffinity labeled molecule with propargyl group 1 has been designed and synthesized, followed it's labeling of N2-acetyl-2'-O-propargyl guanosine 9 by "click chemistry". This technology presents delight development potential in labeling of second messenger cyclic nucleotide, antisense oligonucleotide or siRNA.
Biotin
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chemistry
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Click Chemistry
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Guanosine
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chemical synthesis
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chemistry
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Ligands
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Photoaffinity Labels
4.Studies on preparation by SPG membrane emulsification method and in vitro characterization of tetradrine-tashionone II(A)-PLGA composite microspheres.
Jin LU ; Meng ZHANG ; Hua-xu ZHU ; Li-wei GUO ; Lin-mei PAN ; Ting-ming FU
China Journal of Chinese Materia Medica 2015;40(6):1091-1096
Tetradrine-tashionone II(A)-PLGA composite microspheres were prepared by the SPG membrane emulsification method, and the characterization of tetradrine-tashionone II(A) -PLGA composite microspheres were studied in this experiment. The results of IR, DSC and XRD showed that teradrine and tashionone II(A) in composite microspheres were highly dispersed in the PLGA with amorphous form. The results of tetradrine-tashionone II(A) -PLGA composite microspheres in vitro release experiment showed that the cumulative release amounts of tetradrine and tashionone II(A) were 6.44% and 3.60% in 24 h, and the cumulative release amounts of tetradrine and tashionone II(A) were 89.02% and 21.24% in 17 d. The process of drug in vitro release accorded with the model of Riger-Peppas. Tetradrine-tashionone II(A) -PLGA composite microspheres had slow-release effect, and it could significantly reduce the burst release, prolong the therapeutic time, decrease the dosage of drugs and provide a new idea and method to prepare traditional Chinese medicine compound.
Benzofurans
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chemistry
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Benzylisoquinolines
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chemistry
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Drug Carriers
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chemistry
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Drug Compounding
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instrumentation
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methods
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Drugs, Chinese Herbal
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chemistry
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Kinetics
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Lactic Acid
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chemistry
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Microspheres
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Particle Size
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Polyglycolic Acid
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chemistry
5.Experimental study on formation of neutrophil extracellular traps in human peripheral blood induced by paraquat in vitro
Wei JIN ; Jian LU ; Hui XIE ; Yiqun JIANG ; Xiaoxiao MENG ; Yong ZHU ; Ruilan WANG
Chinese Critical Care Medicine 2017;29(3):216-220
Objective To explore whether paraquat (PQ) can induce the formation of neutrophil extracellular traps (NETs) in human peripheral blood.Methods Neutrophils were isolated from healthy human peripheral blood,and the cells were identified by hematoxylin-eosin (HE) strain.The cells were treated with different concentrations of PQ [0 (as control),200,400,600,800,1 000 and 1 200 μmol/L],and the cell viability was measured by cell proliferation and CCK-8 cytotoxicity detection kit,and the median lethal concentration of PQ was selected.The cells were treated with the median lethal concentration of PQ (PQ poisoning group),and the untreated cells were served as the control.Immunofluorescence staining was adopted to evaluate NETs formation.PicoGreen dye was used to determine the quantitative content of circulating free DNA.Western Blot was used to determine the expressions of citrullinated histone 3(H3Cit) and myeloperoxidase (MPO) in the supernatant.Results The purity of neutrophils was about 95% by HE staining.The cells were treated with different concentrations of PQ,and the result showed that the viability of cells was (58 ± 2)% with 800 μmol/L PQ for treatment.The immunofluorescence showed that there were few expressions of H3Cit and MPO in neutrophils in the control group,and there was no NETs formation,which was composed of DNA,H3Cit and MPO.Compared with the control group,a large amount of NETs was generated from neutrophils stimulated by 800 μmol/L of PQ.Meanwhile,quantitative result showed that the content of cell free DNA in the supernatant was significantly increased in PQ poisoning group as compared with that of control group (μg/L:2 235 ± 462 vs.561 ± 87,P < 0.01).The protein expressions of H3Cit and MPO in the supernatant were also significantly increased as compared with those of control group [H3Cit protein expression (gray value):0.23 ± 0.03 vs.0.11 ± 0.01,MPO protein expression (gray value):0.47 ± 0.05 vs.0.21 ± 0.04,both P < 0.05].Conclusion 800 μmol/L of PQ can induce the formation of NETs in human peripheral blood.
6.Correlation study of the 24-hour ambulatory systolic blood pressure and carotid intima-media thickness in the elderly
Keyu HUANG ; Chunhui LI ; Hualing ZHAO ; Lu SONG ; Yiming WANG ; Hui YANG ; Lingmin MENG ; Ying ZHU
Clinical Medicine of China 2016;32(7):585-589
Objective To investigate the correlation of the 24?hour ambulatory systolic blood pressure (SBP) and carotid intima?media thickness(CIMT) in the elderly. Methods A total of 2 464 who were more than or equal to 60 years old participants were selected with random sampling in accordance with the inclusion criteria from the retired workers in Tangshan Kailuan Company. Dynamic blood pressure monitoring, neck vascular ultrasound and other examination were performed for the participants. . Multivariable linear regression analysis was used to analyze correlation between the SBP of 24?hour, daytime and nightime with CIMT, respectively. Results ( 1) The observation population was divided into three groups according to the tertiles of SBP of 24?hour, daytime and nightime, respectively. With the levels of different SBPs inceasing, CIMT values thickened markedly ( P<0. 01 ) . ( 2 ) Multivariable linear regression analysis showed that after adjusting for confounding factors,the SBP of 24?hour,daytime and nightime associated with CIMT positively and linearly(P<0. 05),and regression coefficient(95%CI) were 0. 022(0. 009-0. 035), 0. 021(0. 008-0. 035), 0. 019 ( 0. 006-0. 032) respectively. In addition,clinic SBP step into the multivariable linear regression,and regression coefficient ( 95%CI ) were 0. 016 ( 0. 003-0. 029 ) , 0. 016 ( 0. 003-0. 030 ) , 0. 019 ( 0. 007-0. 032 ) , respectively. Conclusion The effect of increased 24?hour ambulatory SBP on CIMT was greater than the clinic SBP. Active monitoring of 24 h ambulatory blood pressure and maintaining a low level of blood pressure is essential for preventing and delaying atherosclerosis.
7.Effect of Shoutaiwan on Expression of α-enolase in Decidua Tissue of Recurrent Abortion Mice
Zhanwang TAN ; Lei LEI ; Huifang LI ; Wei ZHU ; Qiong MENG ; Lei LUO ; Lili LU
Chinese Journal of Information on Traditional Chinese Medicine 2014;(1):65-68
Objective To observe the effect of Shoutaiwan on the expression ofα-enolase in the decidua tissue of recurrent abortion mice. Methods The abortion-prone CBA/J × DBA/2 matings were established as the model of recurrent abortion and the nonabortion-prone CBA/J×BALB/C matings were used as the model of normal pregnancy. The model of recurrent abortion CBA/J × DBA/2 of pregnant mice were randomly divided into model group and Shoutaiwan high-, medium-, low-dose groups, pregnant mice of every group were orally administrated in different doses. On the 14th day of pregnancy, the mice were killed. The expression ofα-enolase was detected by using immunohistochemical method and Western Blot. Results α-enolase expression in the model group was significantly higher than the normal pregnancy group (P<0.01). Compared with the model group, Shoutaiwan low-, medium- and high-dose group significantly decreasedα-enolase expression of pregnant mice (P <0.01). Compared with high-dose group, Shoutaiwan medium-and low-dose group showed no significant difference (P>0.05). Conclusion Shoutaiwan could down-regulate the expression ofα-enolase in the decidua tissues of recurrent abortion mice, which may be one of its mechanisms of preventing miscarriage.
8.Identification of novel epithelial ovarian cancer biomarkers by cross-laboratory microarray analysis.
Xuefeng, JIANG ; Tao, ZHU ; Jie, YANG ; Shuang, LI ; Shuangmei, YE ; Shujie, LIAO ; Li, MENG ; Yunping, LU ; Ding, MA
Journal of Huazhong University of Science and Technology (Medical Sciences) 2010;30(3):354-9
The purpose of this study was to pool information in epithelial ovarian cancer by combining studies using Affymetrix expression microarray datasets made at different laboratories to identify novel biomarkers. Epithelial microarray expression information across laboratories was screened and combined after preprocessing raw microarray data, then ANOVA and unpaired T test statistical analysis was performed for identifying differentially expressed genes (DEGs), followed by clustering and pathway analysis for these DEGs. In this work, we performed a combination analysis on microarrays from three different laboratories using gene expression data on ovarian cancer and obtained a list of differential expression profiles identified as potential candidate in aggressiveness of ovarian cancer. The clustering and pathway analysis explored the different molecular basis of different ovarian cancer stages and potential important regulatory pathways in ovarian cancer development. Our results showed that combination of microarray data from different laboratories in the same platforms may overcome biases derived from probe design and technical features, thereby accelerating the identification of trustworthy DEGs, and demonstrating the advantage of integrative analysis in gene expression studies on epithelial ovarian cancer research.
9.Evaluation of the graft hemodynamics after liver transplantation by CT perfusion
Xiaochun MENG ; Kangshun ZHU ; Yan ZOU ; Lingrong PENG ; Lin LUO ; Hong SHAN ; Hua LI ; Yang YANG ; Minqiang LU ; Guihua CHEN
Chinese Journal of Radiology 2008;42(5):455-459
Objective To investigate the influence factors on the graft hemodynamics after liver transplantation by CT perfusion(CTP).Methods Thirty three liver recipients received CT angiography (CTA)and CTP after liver transplantation.The cases would be excluded when their peak values of the aorta enhancement on time-density curves were out of 95%confidence level.The 95% confidence levels of the hepatic artery perfusion(HAP),portal vein perfusion(PVP),total liver perfusion(TLP)and hepatic perfusion index(HPI)were calculated based on the recipients without postoperative complications and named them as references to those with complication.Results Twenty nine recipients were enrolled in the study.15 of them had no postoperative complication while the other 14 had.The 95% confidence levels of HAP,PVP,TLP and HPI on the 15 recipients without complications were(0.1509-0.3183),the 14 cases with complications.HAP decreased in 7 cases,5 of them had hepatic artery stenosis and 3 of them had splenomegaly.HAP increased in 2 cases.both of them had portal vein stenosis.PVP decreased in 13 cases,8 of them had portal vein stenosis,portal vein thrombosis or occlusion,4 of them had splenorenal shunts and 2 of them had fatty liver.TLP decreased in 12 cases and coincident with PVP decreasing.Only 2 cases had HPI decreasing accompanied with HAP decreasing.Conclusion The hepatic blood perfusion through the hepatic artery and portal vein could be quantitatively measured non-invasively by CTP.The severity and the subtypes of the hepatic ischemia could be evaluated objectively,which is helpful for treatment guidance.
10.Obstruction of hepatic vein or inferior vena cava after liver transplantation:the diagnosis and interventional treatment
Kangshun ZHU ; Jiesheng QIAN ; Xiaochun MENG ; Shuhong YI ; Pengfei PANG ; Keke HE ; Zaibo JIANG ; Minqiang LU ; Hong SHAN
Chinese Journal of Radiology 2010;44(4):411-416
Objective To investigate the diagnosis and interventional therapeutic technology for the obstruction of hepatic vein(HV)or inferior vena cava(IVC)after liver transplantation.Methods In the 831 patients who received orthotopic liver transplantation(OLT)and 26 patients who received living donor liver transplantation(LDLT),11 cases were confirmed with HV or IVC obstruction by venography and received interventional treatment from 2 to 111 days after liver transplantation.Of the 11 patients,five had the obstruction of HV anastomosis,five had the obstruction of IVC anastomosis,and one had the obstruction of HV and IVC anastomosis.In the eleven patients,five patients underwent OLT,four patients underwent LDLT,and two pediatric patients underwent reduced-size OLT.Before interventional treatment,9 patients received CT enhanced scans,2 received MR enhanced scans.Follow-up evaluations included liver or renal function tests,clinical symptom,and monitoring of HV or IVC flow.Pressure gradients before and after interventional treatment were compared by using a paired t test.The imaging data and interventional therapeutic technology of 11 cases were retrospectively analyzed.Results In all 11 patients,CT or MRI could clearly show congested areas of the liver,and the location and degree of HV or IVC obstruction.Of the 11 patients,four with HV obstruction and five with IVC obstruction were treated with stent placement,one with HV obstruction was treated with percutaneous transluminal angioplasty(PTA),one with HV and IVC obstruction was treated with HV PTA and IVC stent placement.Interventional technical success was achieved in all patients.The venous pressure gradient across obstruction was significantly reduced from(16.5±4.1)mm Hg(1 mm Hg=0.133 kPa)before the procedure to(2.9±1.7)mm Hg after the rocedure(t= 11.5,P<0.01).Clinical improvement was noted in 10 patients except one pediatric patient who died of multiple-organs failure at the 9 th day after the treatment During the follow-up period of 9 to 672 days,two patients with PTA treatment had recurrent HV stenosis within one month after treatment,no patient with stent placement developed venous restenosis.No major complications occurred during the procedures.Conclusions Stent placement is safe and effective for HV or IVC obstruction after liver transplantation.CT or MRI before treatment is of important value for the diagnosis of congested areas of the liver,and the observation of HV or IVC obstruction.