1.Changes of ICAM-1 expression of myocardial ischemic reperfusion injury in old rats and cardiac protective effect of Esmolol
Chinese Journal of Rehabilitation Theory and Practice 2002;8(7):423-425
ObjectiveTo investigate the changes of expression of intercellular adhesion molecule 1 (ICAM-1) on myocardial ischemic reperfusion injury(IRI) in old rats and the cardiac protective effect of Esmolol(ES). Methods116 rats were divided into three groups: IR group, IR+ES group and Sham group. The ischemic samples were observed in ischemia and 3,6,12,24 hours after IR. The myocardial levels of expression of ICAM-1 mRNA were evaluated by method of IN Situ Hybridization and the protein were evaluated by immunocytochemistry. The content of infiltration of polymorphonuclear neutrophils(PMNs), malomdialdehyde(MDA), superoxides dismutase(SOD) and the myocardial infarction area were measured too. ResultsAfter IR, myocardial levels of expression of ICAM-1 mRNA, protein,MDA and PMNs were increased significantly; SOD was decreased significantly. Between the levels of expression of ICAM-1 protein and PMNs, infarction area of myocardium, a close correlation were observed(P<0.05). In IR+ES group, all of the indicators were increased after IR, but the levels of increase in IR+ES group were more significantly modification as compared with IR group(P<0.05-0.01).Conclusions The findings indicate that PMNs could induce myocardial IRI after IR, which result from the ICAM-1 mediated PMNs adhesion.ES is able to decrease myocardial IRI by blocking the expression of ICAM-1 partially.
3.Establishment and Management of Multicentral Collection Bio-sample Banks of Malignant Tumors from Digestive System.
Si SHEN ; Junwei SHEN ; Liang ZHU ; Chaoqun WU ; Dongliang LI ; Hongyu YU ; Yuanyuan QIU ; Yi ZHOU
Chinese Journal of Medical Instrumentation 2015;39(6):410-414
To establish and manage of multicentral collection bio-sample banks of malignant tumors from digestive system, the paper designed a multicentral management system, established the standard operation procedures (SOPs) and leaded ten hospitals nationwide to collect tumor samples. The biobank has been established for half a year, and has collected 695 samples from patients with digestive system malignant tumor. The clinical data is full and complete, labeled in a unified way and classified to be managed. The clinical and molecular biology researches were based on the biobank, and obtained achievements. The biobank provides a research platform for malignant tumor of digestive system from different regions and of different types.
Biological Specimen Banks
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organization & administration
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Digestive System
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pathology
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Humans
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Neoplasms
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Specimen Handling
4.Proliferation of retinal pigment epithelial cells induced by (R,R)-XY-10 and (S,S)-XY-10 and their action mechanisms
Yu-Wen, CHENG ; Yu-Liang, WANG ; Yi-Hua, ZHANG ; Si-Xun, PENG ; George C Y CHIOU
International Eye Science 2009;9(9):1641-1645
AIM: To investigate the mechanism of proliferation effect induced by (R,R)-XY-10 and (S,S)-XY-10 on retinal pigmented epithelial cells(ARPE-19).METHODS: Human retinal pigmented epithelial cells(ARPE-19) and human umbilical vein endothelial cells (HUVECs) were used to investigate the effect of (R,R)-XY-10 and (S,S)-XY-10 on cell growth,and their mechanisms of proliferative action by using ERK、 AKT、PI3K、Protein kinase C (PKC)and Nitric oxide synthase (NOS) inhibitors.RESULTS: (R,R)-XY-10 and (S,S)-XY-10 dose-dependently increased ARPE-19 cell proliferation,but not on HUVECs. When treated with proliferative inhibitors,H7(5μmol/L)、hypericin(20μmol/L)、PD98059(2μmol/L)、LY294002(50μmol/L)、SH-5 (10μmol/L) and L-NAME (100μmol/L),the proliferative effect was reduced by H7、hypericin、PD98059 and LY294002,but not by SH-5 and L-NAME.CONCLUSION: (R,R)-XY-10 and (S,S)-XY-10 can induce cell proliferation through MAPK and PI3K dependent pathway. KEYWORDS: age-related macular degeneration; (R,R)-XY-10; (S,S)-XY-10; ARPE-19 cells; human umbilical vein endothelial cells; proliferation
5.The influence of E-cadherin/β-catenin on the glycolysis effect in PANC-1 cells
Yi QIN ; Dingkong LIANG ; Si SHI ; Shunrong JI ; Bo ZHANG ; Wenyan XU ; Jiang LIU ; Jin XU ; Quanxing NI ; Xianjun YU
China Oncology 2015;(2):81-86
Background and purpose:Lower expression of E-cadherin is associated with metastasis of cancer cells, however, the correlation between E-cadherin and glucose metabolism has seldom been reported. This article studied the correlation between E-cadherin and glycolysis effect in PANC-1 cells.Methods:Through treatment of transforming growth factor β (TGF-β) in PANC-1 cells to decrease E-cadherin expression, knock-down the gene of E-cadherin interaction protein β-catenin, and overexpressing of E-cadherin, the effects of E-cadherin on the glucose uptake and lactate production ability and on the expression of key glycolytic genes were assessed.Results:E-cadherin negatively regulated the glycolytic effect of PANC-1 cells by inhibiting glucose uptake and lactate production (P<0.05). Moreover, E-cadherin interacting partner β-catenin signiifcantly promoted glucose metabolism transformation in PANC-1 cells (P<0.05). Moreover, key glycolysis regulator sirtuin 3 (SIRT3) could lower E-cadherin expression.Conclusion:Lower expression of E-cadherin induced the transformation of glucose metabolism transformation in PANC-1 cells and manipulation of E-cadherin expression level could change the glycolysis effect. Moreover, through maneuver glycolysis process could inhibit high metastatic potential of pancreatic cancer cells.
6.Influence of pinacidil on the myocardial mitochondrial structure and the respiratory function in scalded rats.
Nian-yin PENG ; Hong ZHOU ; Liang-yi SI
Chinese Journal of Burns 2005;21(3):170-172
OBJECTIVETo investigate the influence of pinacidil preconditioning on the protection of the structure and respiratory function of injured myocardial mitochondria in scalded rats.
METHODSSeventy-five healthy Wistar rats, weighed 250 approximately 300 g, were randomly divided into three groups: i.e. control (n = 9, with intraperitoneal injection of 50 microg/kg isotonic saline), scald (n = 33, with 30% TBSA full thickness scald) and pre-conditioning (n = 33, with same extent of scald injury after intraperitoneal injection of 50 microg/kg pinacidil) groups. Mitochondrial ultrastructure was observed by transmission electron microscope. The mitochondrial respiratory function, the MDA content and the superoxide anion level were determined with corresponding methods.
RESULTSThe degree of injury to rat myocardial mitochondria in pre-conditioning group was less intensive than that in scald group (P < 0.05 or 0.01). The respiratory control rate in pre-conditioning group was obviously higher than that in scald group (P < 0.05), and the contents of MDA and superoxide anion in pre-conditioning group were markedly lower than those in scald group (P < 0.05 or 0.01), as evidenced by their contents at 3 post scalding hours (0.60 +/- 0.09 micromol/g and 0.127 +/- 0.020) were obviously lower than those in scald group (0.83 +/- 0.07 micromol/g and 0.169 +/- 0.015) (P < 0.01).
CONCLUSIONPinacidil preconditioning was beneficial in the protection of myocardial mitochondria in scalded rats, and it might be related to the pre-opening of potassium channel which was sensitive to mitochondrial ATP.
Animals ; Burns ; drug therapy ; metabolism ; pathology ; Cell Respiration ; drug effects ; Disease Models, Animal ; Mitochondria, Heart ; metabolism ; pathology ; Pinacidil ; therapeutic use ; Rats ; Rats, Wistar ; Superoxides ; analysis
7.Construction of Neisseria surface protein A gene vaccine of Neisseria gonorrhoeae and evaluation of the immune responses induced by this vaccine in mice model
Liang-Yi XIE ; Si-Hai HU ; Xiang-Yun TANG ; Sheng-Hui YANG ; Min-Jun YU ; Fulang HAN ;
Chinese Journal of Infectious Diseases 2007;0(07):-
Objective To construct the Neisseria surface protein A (NspA) DNA vaccine of Neisseria gonorrhoeae and evaluate the humoral and cellular immune responses induced by this vaccine in mice model.Methods The recombinant expression vector pcDNA3.1 (+)/NspA was constructed by inserting NspA gene into the eukaryotic expression vector pcDNA3.1 (+) and confirmed by poly merase chain reaction (PCR),restriction enzymes HindⅢ,XbaⅠand DNA sequencing.NspA mR- NA in transfected RAW264.7 cells and NspA protein expression in transfected COS-7 cells were de- tected by reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemical stai- ning,respectively.Forty-five male BALB/c mice were immunized with pcDNA3.1 (+)/NspA recom binant plasmid.The level of serum anti-Neisseria gonorrhoeae antibody of the immunized mice was detected by tube agglutination test,and the level of interieron (IFN)-?was assayed by enzyme-linked immunosorbent assay (ELISA).The proliferation of splenocytes was determined by methyl thiazolyl tetrazolium (MTT) colormetry.The NspA gene in BALB/c mice was identified by PCR with the total DNA extracted from quadriceps femoris in immunized sites.Results Restriction enzymes digestion a- nalysis and DNA sequencing results revealed that the pcDNA3.1 (+)/NspA had been constructed successfully.NspA gene had been transcripted and expressed in mammalian cells.The peak titer of specific antibody was 1:640 in pcDNA3.1(+)/NspA immunized group and there was no specific an- tibody detected in both pcDNA3.1 (+) immunized group and PBS group.The IFN-?level in pcD NA3.1 (+) immunized group was (23.79?11.85)pg/mL and that in pcDNA3.1 (+)/NspA immu- nized group was(169.71?30.52)pg/mL (P
8.Soluble expression and characterization of disulfide bond-rich subdomains of membrane protein p185 in Escherichia coli.
Liang-Wei LI ; Hai-Bo LIU ; Si-Yi HU ; Dun LIANG ; Lian-Sheng CHENG ; Jing LIU
Chinese Journal of Biotechnology 2005;21(4):590-596
Transmembrane protein p185 (the product of Her2/c-erbB-2 gene) is a member of the epidermal growth factor receptor (EGFR) family. Its overexpression was found in about 30% of breast cancer. It is essential to obtain soluble extracellular domain (ECD) of p185, especially disulfide bond rich domains, for identifying the epitopes of anti-p185 antibodies and researching the interrelationship between the antigen and antibody. The disulfide bond rich domain I-II and domain IV of p185 ECD were amplified from plasmid pBabe/erbB-2 by PCR respectively. These two fragments were inserted into pGEX/4T-1 vector, transfected into E. coli Origami B (DE3) pLysS and expressed inductively by low concentration of IPTG and low temperature overnight. After the pressure lysis of cells, the supernatants were analyzed by SDS-PAGE and the result demonstrated that this GST-fusion protein was expressed solubly in the amount of 10-15 mg/L. By the ELISA, Western blot and other immunological assays, the fusion proteins and their GST cut-off derivates both showed binding activities with several anti-p185 antibodies respectively. These results indicated that it was a feasible and effectual method to express disulfide bond rich domain I-II and domain IV of p185 ECD and this method may also be used to express other disulfide bond rich proteins.
Antibodies, Monoclonal
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immunology
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Disulfides
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immunology
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Escherichia coli
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genetics
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metabolism
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Genetic Vectors
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genetics
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Humans
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Receptor, ErbB-2
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biosynthesis
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genetics
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immunology
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Recombinant Fusion Proteins
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biosynthesis
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genetics
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immunology
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Solubility
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Transfection
9.Expression profiling of MicroRNAs in hippocampus of rats following traumatic brain injury.
Ting-Yi, SUN ; Xiao-Rui, CHEN ; Zi-Long, LIU ; Li-Li, ZHAO ; Yong-Xiang, JIANG ; Guo-Qiang, QU ; Rong-Shuai, WANG ; Si-Zhe, HUANG ; Liang, LIU
Journal of Huazhong University of Science and Technology (Medical Sciences) 2014;34(4):548-53
The changes of microRNA expression in rat hippocampus after traumatic brain injury (TBI) were explored. Adult SD rats received a single controlled cortical impact injury, and the ipsilateral hippocampus was harvested for the subsequent microarray assay at three time points after TBI: 1st day, 3rd day and 5th day, respectively. We characterized the microRNA expression profile in rat hippocampus using the microRNA microarray analysis, and further verified microarray results of miR-142-3p and miR-221 using quantitative real-time PCR. Totally 205 microRNAs were identified and up-/down-regulated more than 1.5 times. There were significant changes in 17 microRNAs at all three time points post-TBI. The quantitative real-time PCR results of miR-142-3p and miR-221 indicated good consistency with the results of the microarray method. MicroRNAs altered at different time points post-TBI. MiR-142-3p and miR-221 may be used as potentially biological markers for TBI assessment in forensic practice.
10.Relationship between serum level of caveolin-1 and early neurological deterioration in patients with acute cerebral infarction
liang Yi ZHONG ; rong Rong ZHANG ; yuan Si HUANG ; yue Xin QIN
Journal of Shanghai Jiaotong University(Medical Science) 2017;37(12):1678-1681
Objective·To investigate the relationship between serum level of caveolin-1 (Cav-1) and early neurological deterioration (END) in patients with acute cerebral infarction. Methods·A total of 126 consecutive patients with acute cerebral infarction were recruited from July 2016 to January 2017 in Department of Neurology, the First Affiliated Hospital of Chongqing Medical University. The serum Cav-1 levels of all patients were detected by enzyme-linked immunosorbent assay (ELISA) test. The neurological deficits were assessed by the National Institutes of Health Stroke Scale (NIHSS) and the Glasgow Coma Scale (GCS) at the same time. Compared with the admission baseline NIHSS score, if second motor NIHSS score increased ≥ 1 point or the total NIHSS score increased ≥ 2 points within 3 days after hospitalization, they were classified as END group, otherwise they were classified as non-END group. Multivariable Logistic regression analysis was used to examine the independent predictors of END in the patients. Receiver operating characteristic (ROC) curves were obtained to explore Cav-1 levels in predicting END. Results·Serum Cav-1 levels in END group were significantly higher than those in non-END group [(29.88±19.57) ng/mL vs (16.08±13.37) ng/mL, P=0.000]. Based on the ROC curves, the best cut-off point of serum Cav-1 for predicting END was 16.55 ng/mL. The sensitivity and specificity were 73.33% and 74.07%, respectively. Multivariable Logistic regression analysis showed that Cav-1≥16.55 ng/mL remained an independent predictor of END (OR=4.936, 95%CI 1.608-15.155, P=0.005). Conclusion·Serum Cav-1 is an independent predictor of END in patients with acute cerebral infarction.