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.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.
4.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.
5.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
6.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
7.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.
8.Effects of nandrolone phenylpropionate on ultrastructure of aorta in rats with or without movement training
ping-ping, WU ; wen-jun, YANG ; ping, ZHU ; meng-wei, SUN ; feng-lei, QIAN ; ai-yun, LU ; hong, CHEN
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(07):-
Objective To observe the influence of nandrolone phenylpropionate(NP) on the ultrastructure of aorta in rats with or without movement training,and investigate the side effects of NP on the cardiovascular system. Methods Forty male SD rats were randomly divided into sedentary control group,sedentary+medicine group,exercise control group and exercise+medicine group.For the groups with medical treatment,NP of 10 mg/kg one time every three days was injected into the rats via gluteus for eight weeks.For the exercise groups,rats were trained to run on treadmill five days per week for eight weeks.The aortae were sampled and specimens were obtained for transmission electron microscopy. Results The ultrastructure of aorta was normal in sedentary control group.For sedentary+medicine group,mitochondrial swelling,vacuolated cytoplasm and lysis of endothelial cells were observed,disruption of intercellular conjunctions,widening of subendothelial spaces and furcation and breakage of internal elastic lamina were found,and smooth muscle cells changed into synthesis type.For exercise control group,no obvious morphologic change was observed,except that part of the internal elastic lamina disrupted.In exercise+medicine group,breakage and lysis of endothelial cells were observed,widening of subendothelial spaces and lysis of internal elastic lamina were found,and autophagosome and myelinoid body were seen in smooth muscle cells. Conclusion NP may lead to the impairment of endothelial cells and the change of smooth muscle cells into synthesis type.Exercise with NP administration may result in more severe impairment in vessel wall.
9.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.
10.Role of multislice spiral CT in diagnosis of biliary complications after liver transplantation
Xiaochun MENG ; Kangshun ZHU ; Yan ZOU ; Junwei CHEN ; Pengfei PANG ; Hong SHAN ; Shuhong YI ; Junfeng ZHANG ; Minqiang LU
Chinese Journal of Hepatobiliary Surgery 2010;16(7):484-487
Objective To analyze the role of multislice spiral CT in the diagnosis of biliary com-plications following liver transplantation. Methods Forty-four patients with biliary complications re-ceived tri-phase contrast-enhancement CT examination and cholangiography (CP) within one week af-ter the CT scanning. Using the results of CP as the standard, we investigated the efficacy of multislice spiral CT for each kind of biliary complication. All the analyses for bile duct were based on the images on the late portal venous phase and the reconstruction of images performed with multiplan reformat,Results CP depicted biliary strictures involved in extrahepatic bile duct in 23 cases (including the common bile duct and common hepatic duct), left or right hepatic duct in 24 and intrahepatic bile duct in 27. The sensitivity, specificity, accuracy, positive predictive value, and negative predictive value of the CT examination were 91.3%, 83. 3% , 87. 8%, 87. 5% and 88. 2% for biliary stricture in extrahe-patic bile duct, 83. 3% , 88. 2%, 85. 4%, 90. 9% and 78. 9% for biliary stricture in left or right he-patic duct, 74.1%, 92.7%, 80.5%, 95.2% and 65.0%, for biliary stricture in intrahepatic bile duct, respectively. CT detected intrahepatic biloma in 4 cases and abscess in 2 but CP only detected biloma in 2 cases. The other 4 cases did not detected by CP because of severe biliary strictures which filled with biliary sludge. CP confirmed anastomotic bile leak in 3 cases. In these cases, CT only de-picted the fluid collection in hepatic hilum and abdominal cavity, none of the exact leak site could be detected. CP detected biliary sludge or stones in 33 cases. However, the sensitivity, specificity, accu-racy, positive predictive value and negative predictive value of the CT examination for biliary sludge or stones were 72.7%,100.0%,78.1%,100.0%and 47.6%,respectively.Meanwhile,in 1 patient with diffuse intrahepatic biliary strictures,active biliary bleeding was correctly detected by CT exami-nation and confirmed by hepatic arteriography.Conclusion Tri-phase contrast-enhancement CT exam-ination can be used as a general method for biliary complications after liver transplantation.Besides its benefits in biliary complications,it is of great value for depicting complications involved in hepatic pa-renchyma and acute biliary bleeding in the same examination.