1.Effects of Huoxue huayu decoction on expression of retinal glutamate transporter in acute ocular hypertension of rats
Wei-wei, SHANG ; Da-bo, WANG ; Jiao, FANG ; Cai, ZHANG
Chinese Journal of Experimental Ophthalmology 2011;29(3):239-243
Background Hypertension,ischemia and hypoxia induce the increase of glutamate level in eye tissue and furthermore produce excitatory damage and apoptosis of retinal ganglion cells(RGCs).At present,the study on the protection of traditional Chinese medicine on glutamate-induced retinal excitatory damage is lack.objective Present study was to explore the protective effects of Huoxue huayu decoction,a Chinese herbal recipe,on RGCs in acute ocular hypertension model. Methods Fony-five healthy clean Wistar rats were divided into three groups randomly.The acute ocular hypertension models were established in 40 Wistar rats by injecting the normal saline solution into the anterior chamber to elevate the intraocular pressure for 60 minutes.Huoxue huayu decoction of 4 ml was administered intragastrically once per day in 20 model rats.Other matched 5 normal Wistar rats served as normal control group.The rats were sacrificed on 1,3,7 and 14 days after modeling and the retinas were isolated for the histopathologieal and uhrastructure examination.Expression of glutamate transporter in the retina of acute ocular hypertension and normal rat retina were detected using quantitative real-time polymerase chain reaction.The utilization of animals followed the Association for Research in vision and Ophthalmology. Results After acute ocular hypertension.the retina thickness attenuated and the numbers of RGCs decreased under the light microscope in 1,3,7 and 14 days after modeling in comparison with normal control rats.The degradation of the organelle and edema as well as changes of cell nuclei were seen in model rats under the transmission electron microscopy.The expression of glutamate transporter mRNA in model group and glutamate transporter group was rapidly elevated in the first day (P<0.05),descended at the third days(P<0.01)and returned to the normal level 7 days later(P>0.05).Conclusion Huoxue huayu decoction can protect the retina against RGCs damage in the acute ocular hypertension by elevating the expression of glutamate transporters in retina.
2.Non-autophagic degradation roles of autophagy receptors.
Da-wei WANG ; Bin ZHANG ; Bin LÜ ; Guang-xin WANG
Acta Pharmaceutica Sinica 2016;51(1):1-8
A growing body of evidence has indicated the important role of autophagy receptors in directing ubiquitinated or non-ubiquitinated cargos towards autophagy. Autophagy receptors bind to LC3 (microtubule-associated protein 1 light chain 3) on phagophore and autophagosome membranes, and recognize signals on cargoes in the delivery system of autophagy. However, the diverse domains in the receptor structures determine that their roles would never be limited to autophagy. Up to date, increasing numbers of the receptor proteins have been demonstrated to serve as a molecular link or switch participating in autophagic degradation, apoptosis or cell survival signals. Here, we highlight the non-autophagic roles of these receptor proteins to draw attention to this growing research topic.
Apoptosis
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Autophagy
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Humans
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Microtubule-Associated Proteins
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physiology
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Signal Transduction
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Ubiquitination
4.Prokaryotic soluble expression, purification and function study of LEDGF/p75 protein.
Da-Wei ZHANG ; Hong-Qiu HE ; Shun-Xing GUO
Acta Pharmaceutica Sinica 2014;49(8):1200-1207
HIV-1 integrase (IN) is a key enzyme for the viral replication. The protein-protein interaction (PPI) between HIV-1 IN and a cellular cofactor lens epithelium-derived growth factor (LEDGF/p75) is a validated target for anti-HIV drug discovery. In order to build the platform for screening inhibitor against PPI between IN and LEDGF/p75, the vector containing the LEDGF/p75 protein cDNA was constructed and expressed in Escherichia coli and the function of the LEDGF/p75 protein was assayed. The LGDGF/p75 encoding gene optimized according to the preference codon usage of E. coli, was synthesized and cloned into the expression vector pGEX-4T-1 to form a recombined plasmid, then transformed into host cell E. coli BL21 (DE3). The recombined clones were identified and confirmed by BamH I/Sal I digestion and sequencing, the successfully recombined plasmid in the host cell was induced by IPTG and the condition of the expression was optimized. The expressed protein was purified by the Ni2+ affinity chromatography column and SDS-PAGE was used to analyze the molecular weight and specificity. In addition, ELISA assay was used to analyze the function of the recombinant protein. The recombinant LGDGF/p75 was soluble, and expressed highly and stably in E. coli. The protein was proved to enhance HIV-1 IN strand transfer activity in vitro by ELISA. It will be helpful to build the platform of screening inhibitors against PPI between IN and LEDGF/p75.
Cloning, Molecular
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Escherichia coli
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metabolism
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HIV Integrase
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metabolism
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HIV-1
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physiology
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Humans
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Intercellular Signaling Peptides and Proteins
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biosynthesis
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Protein Binding
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Virus Replication
5.Effects of hydrogen-rich saline on expression of phosphor-p38MAPK during cerebral ischemia-reperfusion in rats
Da LI ; Yanqiu AI ; Feng JIN ; Long HE ; Wei ZHANG
Chinese Journal of Anesthesiology 2015;35(3):370-372
Objective To evaluate the effect of hydrogen-rich saline on the expression of phosphor-p38 mitogen-activated protein kinase (p-p38MAPK) during cerebral ischemia-reperfusion (I/R) in rats.Methods Seventy-two adult male Sprague-Dawley rats,weighing 220-250 g,were randomly divided into 3 groups (n =20 each) using a random number table:sham operation group (group S),I/R group and hydrogen-rich saline group (group I/RH).Cerebral ischemia was induced in chloral hydrate-anesthetized rats by 2 h middle cerebral artery occlusion in I/R and I/RH groups.The artery was only exposed but not occluded in group S.At 3 days before operation and immediately after onset of reperfusion,hydrogen-rich saline (0.6 mmol/L) 10 ml/kg was intraperitoneally injected in group I/RH,while the equal volume of normal saline was given in S and I/R groups.Neurological deficits were blindly assessed and scored at the end of 24 h reperfusion.The animals were then sacrificed,and brains were removed for microscopic examination and for determination of the cerebral infarct size (by TTC),brain water content,cell apoptosis (by TUNEL),and expression of p38MAPk and phosphor-p38MAPK (p-p38MAPK) (by immunohistochemistry and Western blot).Apoptosis index was calculated.Results Compared with group S,neurological deficit score,apoptosis index,brain water content and cerebral infarct size were significantly increased,and the expression of p38MAPK and p-p38MAPK was up-regulated in I/R and I/RH groups.Compared with group I/R,neurological deficit score,apoptosis index,brain water content and cerebral infarct size were significantly decreased,and the expression of p38MAPK and p-p38MAPK was down-regulated in group I/RH.The pathological changes of cerebral tissues were significantly attenuated in group I/RH as compared with group I/R.Conclusion Hydrogen-rich saline can reduce cell apoptosis through inhibiting p-p38MAPK expression,thus attenuating cerebral I/R injury in rats.
6.Construction of eIF-4E Adenovirus and Its Effect on Metastasis Ability of Human Breast Cancer Cell Line MCF-7
Guo-Qiang ZHANG ; Wei PAN ; Liang ZHAO ; Da PANG ;
China Biotechnology 2006;0(03):-
Objective: To reconstruct adenovirus vector of breast eukaryotic initiation factor 4E and to observe its effect on the metastasis ability of breast cancer cell line MCF-7. Methods: eIF-4E gene was constructed into adenovirus vector pAD-X by gene recombination technique, which was transformed into 293 packaged cell for high titer adenovirus. Real-time PCR was applied to detect eIF-4E gene expression. eIF-4E siRNA was applied and then transwell cabin assay was used to observe changes of invasion and motor ability of MCF-7 cells transfected with reconstruction adenovirus. Result:The finding of digestion was coincided with expected. eIF-4E gene over-expression was detected in transfected MCF-7 cells with real-time PCR. And the invasion and motor abilities of transfected MCF-7 cells were more significantly inhibited in transwell cabin assay (respectively p
7.Effect of hydrogen on activation of caspase-3 in brain tissues during cerebral ischemia-reperfusion in rats
Feng JIN ; Long HE ; Yanqiu AI ; Da LI ; Wei ZHANG
Chinese Journal of Anesthesiology 2017;37(3):305-307
Objective To evaluate the effect of hydrogen on the activation of caspase-3 in brain tissues during cerebral ischemia-reperfusion (I/R) in rats.Methods Thirty-six healthy adult male Sprague-Dawley rats,weighing 220-250 g,were divided into 3 groups (n=12 each) using a random number table:sham operation (group S),I/R group and hydrogen group (group H).Cerebral ischemia was induced by occlusion of the middle cerebral artery followed by reperfusion in I/R and H groups.In group H,hydrogen-rich saline 5 ml/kg (0.6 mmol/L) was injected intraperitoneally at 3 days before establishment of the model and immediately after the onset of reperfusion.At 24 h of reperfusion,the rats were sacrificed,and hippocampal tissues were obtained for determination of neuroapoptosis (by TUNEL),apoptotic neuron count and expression of activated caspase-3 (by Western blot).The brain tissues in the ischemic area were obtained and stained with haematoxylin and eosin for examination of the pathological changes.Results Compared with group S,the expression of activated caspase-3 was significantly up-regulated,and the apoptotic neuron count was increased in I/R and H groups (P<0.05).Compared with group I/R,the expression of activated caspase-3 was significantly down-regulated,the apoptotic neuron count was decreased (P<0.05),and the pathological changes of brain tissues were significantly reduced in group H.Conclusion The mechanism by which hydrogen inhibits neuroapoptosis during cerebral I/R is probably related to inhibited activation of caspase-3 in brain tissues of rats.
8.Effect of Ligustrazine on Expression of Platelet-Derived Growth Factor-? Receptor and Extracellular Signal Regulated Kinase in Neonatal Rat Cardiac Myocytes in Vitro Induced by Angiotensin Ⅱ
yin-ping, SUN ; da-wei, ZHANG ; xing, WANG
Journal of Applied Clinical Pediatrics 2006;0(13):-
Objective To investigate the effect of ligustrazine on the expression of platelet-derived growth factor-?(PDGF-?) receptor and extracellular signal regulated kinase(ERK1/2) induced by angiotensin Ⅱ(AngⅡ) in cardiac myocytes,and explore the mechanism of therapeutic.Methods Cultured cardiac myocytes of neonatal rats were treated with 10-7 mol/L AngⅡ as Ang Ⅱ group,10-7 mol/L AngⅡ plus 10 mg/L ligustrazine as ligustrazine group,the normally cultured neonatal rat cardiac myocytes as control group.Protein synthesis was measured by -leucine incorporation,and the expression of PDGF-? receptor and ERK1/2 was detected by Western blot.SPSS 11.0 software was used to analyze the data.Results There were significant differences among 3 groups(F=20.71 P
9.The value of left ventricular Tei Index in evaluating the cardiac function and prognosis of patients with sepsis-induced cardiomyopathy
Da ZHANG ; Caijun WU ; Wei JIANG ; Shubin GUO
Chinese Journal of Emergency Medicine 2017;26(5):577-580
Objective To determine whether left ventricular Tei Index evaluate the cardiac function and prognosis of patients with sepsis-induced cardiomyopathy (SIC).Methods A total of 86 patients with septic shock combined with SIC in the emergency department of Beijing Chaoyang Hospital affiliated to Capital Medical University from July 2014 to June 2016 were recruited and divided into non-survival group (n=35) and survival group (n=51) according to 28-day follow-up.Left ventricular Tei Index, BNP, cTNI and left ventricular ejection fraction within the first 24 h after admisson were detected and compared between the two groups.The correlations of left ventricular Tei Index to BNP, cTNI and ejection fraction were analyzed.The receiver operating characteristic curves (ROC) were constructed to analysize the value of Tei Index in evaluating the cardiac function and prognosis.Results The patientsin the non-survival group had a higher Tei Index compared with that in the survival group [(0.75±0.13) vs.(0.51±0.09), P<0.05].The Tei Index of SIC patients was significantly positively correlated with BNP and cTNI (both P<0.05), and significantly negatively correlated with ejection fraction (P<0.05).The AUC of Tei Index for predicting 28-day mortality in SIC patients was high comapred with that of BNP, cTNI and ejection fraction.Conclusion The left ventricular Tei Index has a reliable value in evaluating the cardiac function and prognosis of patients with SIC.
10.Timing of retinal vein occlusion laser photocoagulation
Wei, ZHANG ; Da-Guang, BI ; Shu-Yan, GU
International Eye Science 2014;(8):1403-1406
AIM:To observe the clinical curative effect of different duration of retinal vein occlusion ( RVO ) by laser photocoagulation treatment, discuss the timing of the RVO laser photocoagulation treatment, provide the basis for clinical choice of RVO photocoagulation treatment time.
METHODS: Retrospective analysis. Line selection retinal laser photocoagulation treatment for 103 cases (103 eyes) with RVO, patients were divided into three groups according to the onset time. In group A (46 eyes), course≤ 1mo, 28 eyes with branch retinal vein occlusion ( BRVO ) , 18 eyes with central retinal vein occlusion ( CRVO);30 eyes were ischemic RVO, 16 eyes were non-ischemic RVO. In group B (38 eyes), 1mo
CONCLUSION:RVO laser photocoagulation in the early intervention treatment can accelerate the retinal hemorrhage, macular edema, absorption, effectively protect the patient's existing vision, improve the long-term vision, and has a certain clinical practical significance.