2.Regulatory effects of CD44 antibody-A3 D8 on IL-3 Rαand downstream PI3 K/Akt pathway in NB4 cells
Ping CHEN ; Qin YUAN ; Xi JIANG ; Juanying WU ; Huifang HUANG
Chinese Pharmacological Bulletin 2014;(11):1526-1529
Aim To investigate the effect of CD44 anti-body-A3 D8 on the expression of IL-3 Rα and down-stream PI3K/Akt in NB4 cells. Methods The ex-pression of IL-3 Rα mRNA was detected by real-time quantitative RT-PCR, the IL-3Rα protein expression and changes of PI3 K/Akt signal pathway in NB4 cells treated with A3D8 were analyzed by Western blot. An-nexin-V-FITC/PI double staining flow cytometry was u-tilized to detect the apoptotic cells. The inhibitor of PI3 K/Akt signaling LY294002 combined with A3 D8 was used to inhibit the PI3K/Akt in NB4 cells. Re-sults After treated with A3 D8 , both the transcription-al level and translational level of IL-3 Rα were remark-ably reduced, and the PI3K/Akt pathway was inhibi-ted. LY294002 improved the inhibitory and apoptotic effects of A3D8 on NB4 cells. Conclusion CD44 antibody A3 D8 can downregulate the expression of IL-3Rα and inhibit the downstream PI3K/Akt pathway.
3.Effects of Rhizoma Acori Tatarinowii on Amino Acids Neurotransmitter in Mice Brain
Hongmei TANG ; Ping XI ; Min WU ; Qingqun CAI
Traditional Chinese Drug Research & Clinical Pharmacology 2000;0(05):-
Objective To study the effects of Rhizoma Acori Tatarinowii (RAT) on amino acids neurotransmitter in mice brain and to explore its mechanisms. Methods After treated by different parts of RAT,the cerebral contents of the amino acids neurotransmitter in rats were detected by high performance liquid chromatography (HPLC) Results Volatile oil, fluid extract and defatted decoction of RAT could significantly lower cerebral glutamic acid level. Volatile oil and fluid extract decreased the content of aspartic acid, and volatile oil could also reduce the taurine content in brain tissue. Conclusion RAT can lower the contents of excitatory amino acids(EAA ) and has protective effect on the brain.
4.Clinical effect of Danshen injection iontophoresis combined with Lumbrokinase in early retinal vein occlusion patients
Xi-Gang, ZHANG ; Ping, WU ; Ying, TONG ; Yin-Lan, LI
International Eye Science 2015;(1):101-103
Abstract? AlM: To explore the effect of Danshen injection iontophoresis combined with lumbrukinase on early patients with retinal vein occlusion ( RVO ) , and observe the prognosis to provide objective basis for clinical?METHODS: Eighty-two patients ( 117 eyes ) with early RVO were randomly selected in our hospital from January, 2011 to April, 2014, 41 cases (59 eyes) of control group treated with Lumbrokinase. Forty-one cases ( 58 eyes) of the observation group, treated with Danshen injection iontophoresis combined with lumbrokinase. The main indexes were the treatment effect, visual acuity and retinal circulation time.?RESULTS: The total effective rate of the observation group was 85. 4% ( 35/41 ), more than that of control group 68. 3% ( 28/41 ), the difference was statistically significant (P<0. 05). The invalid rate of the observation group was 14. 6% (6/41), lower than that of the control group 31. 7% ( 13/41 ), the difference was statistically significant (P<0. 05). The vision of the observation group 4. 8 ~5. 0 was 53. 7% (22/41), more than that of control group 41. 5% ( 17/41 ), the difference was statistically significant ( P < 0. 05 ). The restore vision of the observation group 4. 3 ~ 4. 7 was 31. 7% (13/41), lower than that of the control group 41. 5% ( 17/41 ), the difference was statistically significant ( P<0. 05 ). After treatment, retinal circulation time of two groups were lower than before treatment, the difference was statistically significant (P<0. 05). After treatment of retinal vein circulation time of the observation group was 8. 15 ± 1. 30s, 9. 70 ± 1. 28s lower than that of the control group, the difference was statistically significant (P<0. 05).?CONCLUSlON: There is better effective on Danshen injection iontophoresis combined with Lumbrokinase for early retinal vein occlusion, and can improve the patient's visual acuity, promote the rehabilitation of patients.
5.Establishment and application of a tandem affinity purification system of innate immune regulatory protein PKR
Yuye LI ; Zhaoduan LIANG ; Siyu WU ; Jiong XIE ; Junfang HE ; Minhao WU ; Xi HUANG ; Ping ZHANG
Chinese Journal of Microbiology and Immunology 2011;31(6):487-491
Objective To establish a tandem affinity purification(TAP) system of innate immune-regulatory protein PKR and analyze PKR function, for the future screen and identification of novel PKR-interaction proteins. Methods PKR gene was amplified by PCR, and then cloned into a mammalian expression vector pcTAP-A. Recombinant pcTAP-PKR was transfected into PKR knock-down(PKRkd) HeLa cells by LipofectAMINE 2000,and the PKR overexpressed HeLa cells were harvested for mitogen-activated protein kinases(MAPK) activation analysis. Cell extracts of PKR overexpressed cells were purified using TAP kit and examined by Western blot. Results Cal modulin resin(CBP) and streptavidin resin(SBP) tagged PKR was detected in PKRkd HeLa cells as early as 24 h upon transfection with pcTAP-PKR, and its expression decreased at later time points. The overexpression of PKR was autophosphorylated, and thus involved in the regulation of MAPK actviation. After small-scale TAP kit purification, PKR protein was detectable by Western blot. Conclusion We have successfully established a TAP system that over-expresses functional PKR, providing a useful tool for the future study on the identification of PKR interacting proteins.
6.Inhibitory effect of cinnamaldehyde on invasion capacities of human breast cancer cell line MDA-MB-435S and its relation with regulating the expression of miR-27a.
Rui-Ping WANG ; Ge WANG ; Qing-Min SUN ; Jian WU ; Xi ZOU
Chinese Journal of Integrated Traditional and Western Medicine 2014;34(8):964-969
OBJECTIVETo explore the inhibitory effect of cinnamaldehyde on invasion capacities of human breast cancer cell line MDA-MB-435S and its relation with regulating the expression of miR-27a.
METHODSThe effect of cinnamaldehyde on invasive capacities of MDA-MB-435S was measured by Transwell matrigel invasion assay. The effect of miR-27a expression on invasive capabilities of MDA-MB-435S, the intervention of cinnamaldehyde in the miR-27a expression, and its relation with its effect on invasive capabilities were defected with liposome 2000 transinfection miRNA27a mimics/inhibitors, real time-polymerase chain reaction (Real-time PCR), and Transwell chamber model.
RESULTSCompared with the control group, the number of cells passing through the transwell chamber was more significantly reduced after treated by cinnamaldehyde for 12 h (P < 0.05). The miR-27a expression was 962.07 times and 40% of that of the control group after transinfected by miR-27a mimics and miR-27a inhibitors. After transinfected by miR-27a inhibitors, the number of cells passing through the transwell chamber was more significantly reduced (P < 0.05). The miR-27a expression of MDA-MB-435S was down-regulated by 12-h treatment of cinnamaldehyde (2(-deltaCt) = 0.56, 0.18, 0.18, respectively). The number of miR-27a mimics transinfection pretreated MDA-MB-435S cells passing through the transwell chamber increased more obviously than the number of un-pretreated MDA-MB-435S cells in the control group (P < 0.05).
CONCLUSIONSCinnamaldehyde could inhibit invasive capabilities of human breast cancer cell line MDA-MB-435S. The over-expression of miR-27a played an important role in the invasive capability of MDA-MB-435S. The inhibition of cinnamaldehyde on invasive capabilities of MDA-MB-435S cells was correlated with down-regulating the expression of miR-27a.
Acrolein ; analogs & derivatives ; pharmacology ; Breast Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Female ; Humans ; MicroRNAs ; genetics
7.Changes of Intestinal Mucosal Barrier and Intestinal Flora in Rats with Severe Acute Pancreatitis.
Yan LI ; Hao WU ; Yiyun DENG ; Ruyi LIAO ; Lili XI ; Ping YAO
Journal of Biomedical Engineering 2015;32(2):412-417
This paper is to explore changes of intestinal mucosal barrier, intestinal flora, and bacterial translocation in rats with severe acute pancreatitis (SAP). Twenty four male SD rats were randomly divided into the control group (n = 10) and the experimental group (n = 14). The model of severe acute pancreatitis of rats was induced by the method of injecting adversely 5% sodium taurocholate into the common biliary-pancreatic duct. All of the rats were killed after 24 hours and the level of the serum amylase and the plasma endotoxin was determined after that. The pathological changes of pancreas and small intestine were observed through hematoxylin-eosin staining (HE staining) and the abdominal viscera bacterial translocation rates were tested. With the method of real-time polymerase chain reaction (RT-PCR) the quantity of the intestinal flora was analyzed. In the control group, the level of Escherichia coli, Lactobacillus and Bifidobacterium were 2.08 ± 1.29, 11.04 ± 7.55 and 12.21 ± 4.95, respectively. On the contrast, the level of Escherichia coli in the cecum contents was much higher (9.72 ± 3.58, P < 0.01), while the Lactobacillus number was decreased significantly (0.67 ± 0.34, P < 0.01), and the Bifidobacterium number was also decreased (4.59 ± 3.42, P < 0.05) in the experimental group, so the ratio of Bifidobacterium/Escherichia coli was reversed. Besides, in the experimental group, the plasma endotoxin positive rates and the bacterial translocation rates were much higher (P < 0.01 or P < 0.05) and the pathology scores of pancreas and small intestines were also significantly higher (P < 0.01) than those in the control group. These results indicated that in severe acute pancreatitis rats, the intestinal mucosal barrier was severely damaged and the dysbacteriosis occurs in the intestinal canal. And these might relate to the occurrence and development of multiple organ infection.
Animals
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Bacterial Translocation
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Endotoxins
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Intestinal Mucosa
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pathology
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Intestines
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microbiology
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Male
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Pancreas
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pathology
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Pancreatitis
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microbiology
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pathology
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Rats
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Rats, Sprague-Dawley
8.A simple and economic method for isolating hepatic lipocytes
Yi-Jun ZHANG ; Xi-Shan YANG ; Ping-Sheng WU ; Shu-Ren LIU
Journal of Southern Medical University 2001;21(4):304-306
Objective To establish a simple and economic method for isolating rat hepatic lipocytes. Method Perfusion of the rat liver with pronase E and collagenase in succession was performed in cycles, and the hepatic lipocytes were isolated and purified by digesting the rat liver with DNase in the presence of the above 3 enzymes, followed by 11% Necodenz density gradient centrifugation. The resulted lipocytes were subjected to routine identification procedures and viability assessment. Results The yield of the lipocytes from each rat was 2×107 to 3×107, with viability over 95% and purity above 90%. Conclusion The method adopted in this study is simple, economic and highly productive.
9.Impact of traumatic events of work on the level of post-traumatic stress disorder among emergency nurses
Lin LIN ; Yi-Xi WU ; Guo-Ping HE
Chinese Journal of Modern Nursing 2009;15(5):407-410
Objective To identify the level of Post-traumatic Stress Disorder(PTSD) among emergency nurses and its relationship to exposure to traumatic events in work. Methods The questionnaire and standardised measures were used to investigate the level of PTSD in the third class of general hospital of Changsha and the effect of the traumatic events on it was analysed. Results 38 out of the 193 objects were detected to be positive PTSD, and the positive rate was 19.7%. Spearman correlation analysis showed that there were positive correlations between PTSD and traumatic events (P < 0. 01). Condusions It was higher that the level of PTSD of emergency nurses in the third class of general hospital of Changsha, and traumatic events of work was the most important reason that resulted in PTSD.
10.A simple and economic method for isolating hepatic lipocytes
Yi-Jun ZHANG ; Xi-Shan YANG ; Ping-Sheng WU ; Shu-Ren LIU
Journal of Southern Medical University 2001;21(4):304-306
Objective To establish a simple and economic method for isolating rat hepatic lipocytes. Method Perfusion of the rat liver with pronase E and collagenase in succession was performed in cycles, and the hepatic lipocytes were isolated and purified by digesting the rat liver with DNase in the presence of the above 3 enzymes, followed by 11% Necodenz density gradient centrifugation. The resulted lipocytes were subjected to routine identification procedures and viability assessment. Results The yield of the lipocytes from each rat was 2×107 to 3×107, with viability over 95% and purity above 90%. Conclusion The method adopted in this study is simple, economic and highly productive.