1.Expressions of GABARB2 in brain regions of rats with high-level and low-level aggressive behavior
Jie GAO ; Sheng WEI ; Mingqi QIAO
Chinese Journal of Behavioral Medicine and Brain Science 2012;21(7):624-626
ObjectiveTo compare the mRNA and protein expression levels ofγ-aminobutyric acid receptor B2 ( GABABR2 ) in brain regions of male rats with high level aggressive behaviors and low level aggressive behaviors respectively,and provide clues for exploring mechanism of GABA in aggressive behaviors.MethodsWistar male rats were randomly divided into two groups:the normal group and the aggressive behavior group.Then social isolation and resident intruder stresses were used to establish high level and low level aggressive behavior in the aggressive behavior group.The mRNA and protein level of GABABR2 in parietal cortex,prefrontal cortex,hypothalamus and hippocampus of the three groups rats ( n=10 in each group) were detected using RT-PCR and Western blot respectively.ResultsThe GABABRB22 RT-PCR/Western blot relative integrated optical density of parietal cortex,prefrontal cortex,hypothalamus and hippocampus in the normal group rats respectively were.Those of the above four brain regions in high aggressive behavior group rats respectively were ( 0.507 ± 0.049/0.626 ±0.038 ),(0.609 ± 0.049/0.652 ± 0.010 ),( 0.359 ± 0.030/0.731 ± 0.044 ) and ( 0.296 ± 0.054/0.452 ±0.079) were significantly lower (P<0.05) compared with the normal group rats.In the low aggressive behavior group rats,the GABABRB2 RT-PCR/Western blot relative integrated optical density of parietal cortex and hippocampus increased statistically(P< 0.05 ),while those of prefrontal cortex and hypothalamus decreased obviously (P < 0.05 ).all in comparison with the normal group rats.Conclusion Different expression levels of GABABR2 in parietal cortex,prefrontal cortex,hypothalamus and hippocampus are relative to aggressive behaviors,which might be one of the mechanism for GABA in aggressive behaviors.
2.The current situation of clinical utilization of biliary stents
Jie GAO ; Wenhua ZHAO ; Kai CUI ; Sheng LI
International Journal of Surgery 2011;38(5):325-328
Biliary stent has been utilized in the patients of benign and malignant biliary strictures and biliary fistula who are unsuitable or unwilling to operate.Its short-term curative effect is distinct,while the long-term effect is relatively poor,because the long-term application is affected by stent blocking and biliary ducts restenosis.Therefore,the research and development of new biliary stents,such as biodegradable stent and covered stent,are the main trend of the current development,which will be of profound significance for the biliary tract disease.
4.Research progress of circulating tumor cells detection and its clinical application
Jie GAO ; Fachang YU ; Kai CUI ; Bo ZHANG ; Sheng LI
International Journal of Surgery 2013;(5):327-331
Cancer cell exfoliation,invasion and entry into circulation system is the early event with metastasis,which provide the possibility to formation of clinical metastase.Further research about the circulating cancer cells can help us to understand the mechanism of metastasis and offer the scientific proof against anti-metastasis.The detection of circulating tumor cells and clinical significance were reviewed.
5.The security of epilepsy patients use both donepezil and transcranial magnetic stimulation
Rui JIAO ; Xinyan JIA ; Xu YUAN ; Jie GAO ; Li SHE ; Wenfang GAO ; Sheng OU ; Shufang YU
Journal of Chinese Physician 2016;(z1):242-244
Epilepsy is a complications of brain injury or stroke,and is a common diseases in reha-bilitation or neurology department.Transcranial magnetic stimulation as a classical treatment means for stroke or brain injury,but also can promote the recovery of epilepsy.However,there is no clear clinical re-port for safety of epilepsy patients use both donepezil and transcranial magnetic stimulation .The article re-views the literature.Clinicians maybe provided some help.
6.Comparative proteomics analysis of aging rat aorta during the process of increasing age
Yuping SHENG ; Yan WANG ; Shaohua ZHAO ; Xiang JI ; Haiqing GAO ; Jie QIU
Chinese Journal of Geriatrics 2013;(1):91-95
Objective To study the proteins related to aging in aortic of old rats for laying the foundation of further study of aging mechanism.Methods The rat model of aging was built,and all model rats were divided into 4 groups:the adult group(9 weeks),the old group(12 months)of WistarKyoto (WKY) rats,and the age-matched spontaneously hypertensive rats (SHR).Blood pressure of 4 groups was observed.Morphological change of aorta was observed by HE staining.Differential proteins were identified by isobaric tags for relative and absolute quantitation,(iTRAQ)-coupled liquid chromatography and tandem mass spectrometry technology.Part of differential proteins was subsequently detected by real time PCR and Western blot.Results The mean SBP of the old group SHR was higher than WKY of 97.1% (t=39.00,P<0.05),and the adult group of SHR was also higher than WKY of 5.4%(t=3.64,P<0.05).Compared with the adult group,aging change in the aortic morphology of old SHR and WKY were shown in HE staining,and the change in SHR rats was more marked.7 proteins related to aging were identified by Mass spectrum analysis,and they were Profilin-1,Prelamin A,HSP70,creatine kinase-M,Fibulin-5,eIF5A and Prohibitin.Part of differential proteins was subsequently confirmed by real time PCR and Western blot.Prelamin-A was up-regulated in the old group of WKY and SHR (0.15±0.01 vs.0.45±0.04,0.34±0.02 vs.0.78±0.06) (t=12.67,12.06,all P<0.01),Prohibitin was down-regulated in the old group of WKY and SHR(1.34±0.05 vs.1.01± 0.06,1.24±0.05 vs.0.88±0.08) (t=7.41,7.09,all P<0.01).Profilin-1 was up-regulated in the old group of WKY and SHR (9.12±0.4 vs.20.76±0.8,16.84±0.5 vs.55.16±0.9) (t=22.55,64.46,both P<0.01),and Profilin-1 expression in the old group of SHR was higher thanWKY (55.16±0.90 vs.20.76±0.8,t=49.49,P<0.01).Conclusions Differential proteins of the old rat aorta are identified through the comparative proteomics method.These differential proteins will provide new targets for the prevention and control of vascular aging.
7.Smac/DIABLO induces apoptosis in pancreatic cancer cells and increases the sensitivity of TRAIL and gemcitabine chemotherapy
Yanling ZHENG ; Yan LI ; Hongmin LUO ; Jie GAO ; He TIAN ; Zuoxing NIU ; Sheng LI
Chinese Journal of General Surgery 2013;(3):211-214
Objective To explore the effect of ectopic overexpression of Smac/DIABLO on the proliferation of pancreatic cancer cell line SW1990,and the sensitization to TRAIL and Gemcitabine induced apoptosis.Methods The Smac/DIABLO gene was transfected into the pancreatic cancer cell line SW1990 with the participation of Lipofectamine 2000 (SW1990/Smac).The cell line transfected with empty vector served as controls (SW1990/neo).The SW1990/neo and SW1990/Smac cells were assigned into the following treatment groups:TRAIL group,Gemcitabine group,TRAIL plus Gemcitabine group,and the control group.The SW1990 cells were treated with TRAIL and Gemcitabine in different concentrations and time.The cell growth inhibition rate (CGIR) was detected by MTT,the rate of apoptosis was measured by flow eytometry,the apoptosis morphous was observed by Heochst 33342 staining.The expressions of apoptosis-associated proteins such as Smas/DIABLO,XIAP,cytochrome C and caspase-3 were detected by Western blot.Results The cell growth of SW1990/Smac was significantly lower than growth of SW1990/ neo.The concentration of TRAIL were 200,500,1000 and 2500 ng/ml respectively.After 24 hours,the CGIR of SW1990/neo and SW1990/Smac were 11.11%,46.03%,67.08%,76.19% and 22.11%,42.67%,56.63%,67.6% respectively (P < 0.05).The concentration of Gemcitabine were 10,20,40 and 60 μmol/L respectively.After 24 hours,the CGIR of SW1990/neo and SW1990/Smac were 15.2%,34.6%,55.16%,76.4% and 22.65%,36.85%,55.11%,79.99% respectively (P<0.05).The cells of SW1990/neo and SW1990/Smac were treated by TRAIL(500 ng/ml),Gemcitabine (20 μmol/L) and combination group.The apoptosis rate were 5.64%,15.30%,27.27% and 20.37%,23.27%,67.30% (P < 0.05) respectively.In combination group,the expressions of activators of caspase such as Smas/DIABLO,cytochrome C and caspase-3 increased significantly,while the expressions of inhibitor of apoptosis protein XIAP decreased.Conclusions Ectopic expression of Smac/DIABLO could induce the apoptosis of SW1990 cell,inhibit the cell proliferation,and enhence the sensitivity of SW1990 cell to TRAIL and Gemcitabine.The mechanism of apoptosis sensitization effect by Smac/DIABLO was associated with significant up-regulation of Smac/DIABLO,cytochrome C,down-regulation of XIAP,and the activation of caspase-3.
9.Effect of hypoxia and hyperglycemia on retinal pigment epithelial cells co-cultured with human mesenchymal stem cells
Jie-Jing, YAN ; Hai-Yan, WANG ; Yu-Sheng, WANG ; Fan, GAO ; Na, LI ; Peng, ZHANG
International Eye Science 2015;(4):584-587
AIM: To evaluate cell proliferation, apoptosis and migration of human retinal pigment epithelial cells ( RPE) when co - cultured with human marrow mesenchymal stem cells ( hMSCs ) in condition of hypoxia and hyperglycemia so as to explore possible mechanisms of diabetes aggravating choroidal neovascularization ( CNV) preliminarily.
METHODS:Both hMSCs and RPE cells were co-cultured in a transwell system. The experiment was divided into four groups: 21% O2 with 5. 56mmol/L glucose ( control group, A ), 21% O2 with 30mmol/L glucose ( hyperglycemia and normoxia group, B ) , 5% O2 with 5.56mmol/L glucose ( normoglycemia and hypoxia group, C ) and 5% O2 with 30mmol/L glucose ( hyperglycemia and hypoxia group, D) . Cell Counting Kit-8 (CCK-8) was used to detect the proliferation of RPE cells in each group at 12, 24 and 48h respectively. Flow cytometry was performed to observe apoptosis of RPE cells at 24h. Additionally, we assessed migration
capabilities of RPE via transwell assay under the condition of hyperglycemia and hypoxia by co-culturing of hMSCs.RESULTS:In this co-culturing system, at 12, 24 and 48h, group B (1. 61±0. 41, 1. 80±0. 34;1. 91±0. 35), C (1.34±0. 46, 1. 94±0. 40, 2. 14±0. 41) and D (1. 98±0. 47, 2.26±0.42, 2. 55±0. 40) showed significantly higher proliferation rate than group A (0. 92±0. 45, 1. 27±0. 32, 1.59±0. 41, P<0. 05). The migration capabilities of RPE in group B (149. 5±9. 19), C (140±9. 90) and D (170. 5±7. 78) increased dramatically compared with group A ( 114. 5±7.78, P<0.05) at 24h, whereas there was no significant difference of apoptosis ratio among four groups (P>0. 05).
CONCLUSION:By coexistence with hMSCs, the synergy of hyperglycemia and hypoxia can improve migration and proliferation of RPE cells, and have no effect on apoptosis of RPE cells within short period.
10.Expression, purification of Staphylococcus aureus Efb protein and preparation of its functional antibody
Xin, ZHANG ; Ya-Ping, GAO ; Jie, DONG ; Yu, LIU ; Na, WU ; Ning-Sheng, SHAO ; Guang, YANG
Bulletin of The Academy of Military Medical Sciences 2009;33(6):531-534
Objective:To clone and express the Staphylococcus aureus Efb(extracellular fibrinogen-binding protein) protein in Escherichia coli, to purify the expression product and prepare its functional antibody and to detect the functions of Efb protein for further studies on S.aureus infection.Methods: Efb gene was amplified by PCR using S.aureus NCTC-8325 genome DNA as template and cloned into the recombinant expression vectors pET28a. E.coli BL21(DE3) with the plasmid was induced with IPTG for protein production. The protein was purified by Ni~(2+) affinity chromatography. The function of Efb protein was determined by complement activity assay and inhibition ELISA.The polyclonal antibodies were prepared by immunizing the animals. Results: The purified recombinant Efb was obtained, which could inhibit the CH50 and AH50 effectively. The functional poly-antibodies of Efb were prepared.Conclusion:Efb could inhibit the classical pathway and alternative pathway of complement activation, and the antibodies against to Efb could block the inhibition of the classical pathway of complement activation induced by Efb.