1.In vitro studies of Raf-CREB, Akt-CREB, and CaMK II -CREB signal transduction pathway regulated by ginsenosides Rb1, Rg1 and Re.
Ting-Ting WANG ; Xian-Zhe DONG ; Wan-Wan LIU ; Yi-Hong CHEN ; Ping LIU
China Journal of Chinese Materia Medica 2014;39(11):2065-2070
OBJECTIVEEffects of ginsenoside Rb1, Rg1 and Re on neurotrophic factor signal transduction pathway using liposome-mediated transfection of eukaryotic cells approach.
METHODThe injury model was established by treating SH-SY5Y cells with 0.6 mmol x L(-1) of corticosterone (CORT) by 24 h. SH-SY5Y cell were pretreated with CORT for 30 min followed by co-treated with 120,60 and 20 micromol x L(-1) of Rb1, 120, 80 and 40 micromol x L(-1) of Rg1 and 120, 80 and 40 micromol x L(-1) of Re for 24 h. Cells viability was determined by Cell Counting Kit (CCK) assay. CREB expressing Luciferase reporter gene was constructed and transfected with plasmid containing hRaf, hcAMP, hAkt, hCaMK gene into human embryonic kidney (HEK293) cells using liposornal transfection reagent lipofection 2000. The expression of CREB before and after it addion of Rb1, Rg1 and Re was examined by Luc assay system and Western blotting.
RESULTCompared with normal control group, CORT significantly decreased the viability of SH-SY5Y cells to 67.21% (P < 0.01). CCK results show that Rb1 (60 micromol x L(-1)), Rg1 (80 micromol x L(-1)) and Re (80 micromol x L(-1)) on SH-SY5Y cells have significant protective effect (P < 0.01). Lucassay and Western blotting results show that the gene and protein levels of CREB increased significantly through the pathway of Raf and Akt with Rb1 and Rg1 (P < 0.01), Re can increase significantly the gene and protein levels of CREB through the pathway of Raf and CaMK II.
CONCLUSIONRb1, Rg1 and Re protects SH-SY5Y cells from CORT-induced damage and the neuroprotective mechanism may be associated with the Raf-CREB, Akt-CREB and CaMK II -CREB pathways.
Calcium-Calmodulin-Dependent Protein Kinase Type 2 ; genetics ; metabolism ; Cell Line ; Cell Survival ; drug effects ; Cyclic AMP Response Element-Binding Protein ; genetics ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Genes, Reporter ; Ginsenosides ; pharmacology ; Humans ; Panax ; chemistry ; Proto-Oncogene Proteins c-akt ; genetics ; metabolism ; Signal Transduction ; drug effects ; raf Kinases ; genetics ; metabolism
3.Biomechanical study of neotype shape memory alloy cervical hook for atlantoaxial instability
Xiaoqiu SHU ; Lei WAN ; Dong YIN ; Chenglong LIU ; Anmin JIN
Chinese Journal of Tissue Engineering Research 2008;12(39):7764-7767
BAcKGROUND:In recent years,the pedicle screws fixation technique,which is used in fixation for atlantoaxial instability associated with trauma,severe degeneration and tumorectomy,has been developed.However,this kind of technique easily causes several complications,including malpositional screws,vascular injuries,and even vertebral artery injury.Based on the biomechanical characteristics of memory alloy and determination of atlantoaxial data,a neotype shape memory alloy cervical hook was designed to treat atlantoaxial instability.OBJECTIVE:To investigate the biomechanieal characteristics of the neotype shape memory alloy cervical hook for atlantoaxial instability.DESIGN,TIME AND SETTING:Repeated measurement analysis of variance test was performed in the Laboratory of Clinical Anatomy and Medical Biomechanics,Southern Medical University between March and April 2008.MATERIALS:Eight fresh adult craniocervical specimens(C0-C4)were provided by Department of Clinical Anatomy,Southern Medical University.Atlantoaxial neotype shape memory alloy cervical hook(50.8%-51.8%nickel and the remaining part was titanium)was fabricated by Shanghai Xinchang Memory Alloy Co.,Ltd.METHODS:The included eight C0-C4 specimens were used to test three-dimension ranges of motion(ROM)by fixation and neotype shape memory alloy cervical hook fixation.Then,the positions of spine varying from no loading to the maximum loading status were scanned and analyzed using image processing software to determine the three-dimensional ROM under different statuses.MAIN OUTCOME MEASURES:Three-dimensional ROM of tested specimens.RESULTS:Neotype shape memory alloy cervical hook fixation and Germany AESCULAP SSE hanger fixation had similar flexion-extension range of motion(P=0.595).Lateral bending three-dimensional ROM was greater in the neotype shape memory alloy cervical hook fixation group than in the Germany AESCULAP AAE hanger fixation(P< 0.05).The rotatory three-dimensional ROM was smaller in the neotype shape memory alloy cervical hook fixation group than in the Germany AESCULAP AAE hanger fixation(P<0.05).CONCLUSION:Neotype shape memory alloy cervical hook fixation had comparative post-surgery immediate stability with the Germany AESCULAP AAE hanger fixation.Neotype shape memory alloy cervical hook fixation produced a little worse biomechanical lateral bending stability and a little better biomechanical rotatory stability than Germany AESCULAP AAE hanger fixation.
4.Expression of heranase and perlecan in the retina of mouse with oxygen-induced retinopathy
Lei, WAN ; Xiaoguang DONG ; Ting, LIU ; Jun, CHENG ; Lixin, XIE
Chinese Ophthalmic Research 2010;28(3):210-215
Background Heparanase degrade heparan sulfate side chains of heparan sulfate proteoglycans in the extracellular matrix.Heparanase induces angiogenesis and likely promotes the vascularization of tumor.ObjectiveThe present study is to investigate the expression of heparanase and perlecan in retinas with oxygen-induced retinopathy.Methods Sixty-five clean neonatal C57BL/6J mice were raised in a hyperbaric oxygen box with a volume percentage of 75%±2% for 5 days and then returned to the normal air room.Another 65 matched mice were raised in the normal environment as controls.Evans blue was infused by the superior vena cava in all the mice on postnatal days 12,13,17,21 and 30,afterwards fluorescein angiography was performed and then the mice were sacrificed.The retinas of mice were isolated and prepared and the retinal vessels were examined under a fluorescent microscope and optical microscope.Heranase and perlecan mRNA was detected using reverse transcription PCR (RT-PCR).Heranase and perlecan proteins were detected by Western blot.The analysis of variance was used to compare the mRNA and the protein levels of heranase and perlecan between the experimental and control groups.Results The expression of heparanase mRNA in the retinas of different ages of mice and the different groups showed significant differences (F_(group)=16.303,P=0.000;F_(time)=18.614,P=0.000;F_(interaction)=11.299,P=0.000),and the expression of heparanase mRNA was significantly enhanced in mice from postnatal days 12,13,17 and 21 compared with normal control mice (P=0.001,0.000,0.000,0.001,respectively).The expression of heparanase protein in the retinas of different ages of mice and the different groups followed the same tendency(F_(group)=458.134,P=0.000;F_(time)=78.466,P=0.000;F_(interaction)=71.398,P=0.000).The expression of perlecan mRNA in the retinas of different ages of mice and the different groups showed significant differences (F_(group)=7.351,P=0.013;F_(time)=9.098,P=0.000;F_(interaction)=3.349,P=0.000),and increase in differences also were clearly seen in mice from postnatal days 13,17 and 21 compared with normal control mice (P=0.048,0.000,0.003,respectively).Conclusion The expression of heparanase and perlecan is associated with the development and progression of retinal neovascularization,and perlecan and heparanase together produce a synergistic effect.Heparanase and perlecan may participate in the angiogenesis of oxygen-induced retinopathy.
5.Cellular immunoregulation of Traditonal Chinese Medicine Huangqi in treating patients with chronic hepatitis B by intravenous injection
Jun DONG ; Jianyong LIU ; Mobin WAN ; Xiantuan TANG ; Juzhen ZHU ;
Academic Journal of Second Military Medical University 2000;0(07):-
Objective: To study the cellular immunoregulatory effect of Huangqi through intravenous injection in patients with chronic hepatitis B. Methods: In treated groups, light and mild chronic hepatitis B (15 patients in each group) were treated with Huangqi and Ganlixin. In control groups, 10 patients in each group with light and mild chronic hepatitis B were treated with Ganlixin. Normal control group included 10 healthy volunteers. Results: Compared with control group, the level of CD3 +, CD4 + and CD4 + /CD8 + in treatment group increased significantly ( P
6.Identification of the metabolites of Dingzhi Xiaowan extract in depressive rat plasma, urine, feces and bile after intragastric administration.
Lu XU ; Wan-wan LIU ; Xiao TAN ; Shi WANG ; Li-hua MU ; Xian-zhe DONG ; Dong-xiao WANG ; Ping LIU
China Journal of Chinese Materia Medica 2015;40(11):2214-2220
Dingzhi Xiaowan is a widely used traditional Chinese medicine in treating depression, which is a similar formula of Kaixinsan. In this research, a rapid ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS(E)) method was established to analyze the metabolites of Dingzhi Xiaowan in depressive model rat plasma, bile, urine and feces. After we established Chronic unpredictable mild stress (CUMS) model rats and orally administrated Dingzhi Xiaowan, rat plasma, bile, urine and feces samples were collected and prepared. Using Waters Cortects UPLC C18 column (2.1 mm x 50 mm, 1.6 μm), acetonitrile-0.1% formic acid mobile phase gradient, these samples were analyzed and 33 metabolites of nine bioactive compounds were detected and tentatively identified by Metabolynx. Among the 33 metabolites, three metabolites were identified from plasma sample, three came from bile sample, and 27 metabolites were identified from urine and feces samples. This approach provided a rapid method for characterizing the metabolites of Dingzhi Xiaowan and gave the truly active structures and the action mechanism of their antidepressant effects.
Animals
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Bile
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metabolism
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Chromatography, High Pressure Liquid
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Depression
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metabolism
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Disease Models, Animal
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Drugs, Chinese Herbal
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metabolism
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Feces
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chemistry
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Male
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Mass Spectrometry
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Medicine, Chinese Traditional
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Plant Extracts
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metabolism
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Rats
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Rats, Sprague-Dawley
7.Effects of Kaixin San formulas on behavioristics and central monoamine neurotransmitters of chronic stress rats.
Wan-wan LIU ; Lu XU ; Xian-zhe DONG ; Xiao TAN ; Shi WANG ; Wei-yu ZHU ; Ping LIU
China Journal of Chinese Materia Medica 2015;40(11):2180-2185
The efficacy of Chinese herbal formula in treating depression has been proved in many studies. In this study, six different Kaixin San formulas were compared to investigate their effects on central monoamine neurotransmitters of chronic stress rats and against depression based on their different components in plasma, in order to discuss the efficacy-comparability relationship and the possible efficacy mechanism. The classic isolation method and the chronic unpredictable mild stress (CUMS) depression model were combined to investigate the changes in contents in hippocampus and monoamine neurotransmitters (NE, DA, 5-HT) and the components of some formulas in plasma with HPLC and UPLC-Q-TOF-MSE methods. As a result, Dingzhi Xiaowan recorded in Essential Recipes for Emergent Use Worth A Thousand significantly increased the behavioral scores, NE and 5-HT contents in hippocampus and NE, DA and 5-HT contents in cortex, with the best anti-depressant effect. Dingzhi Xiaowan recorded in Complete Records of Ancient and Modern Medical Works showed a notable increase in sucrose preference and open field score in model rats, NE content in hippocampus and NE, DA and 5-HT contents in cortex, with a certain anti anti-depressant effect. Kaixin San recorded in Ishinpo showed remarkable rise in weight of model rats. NE content in hippocampus and DA content in cortex. Puxin Decoction recorded in A Supplement to Recipes Worth A Thousand Gold showed 5-HT content in hippocampus and DA content in cortex. Kaixin San recorded in Yimenfang only showed DA content in cortex. Kaixin Wan recorded in Essential Recipes for Emergent Use Worth A Thousand did not mention the antidepressant effect. According to the results, the formulas' different anti-depressant effects may be related to the different plasma components.
Animals
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Behavior, Animal
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drug effects
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Biogenic Monoamines
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analysis
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Brain Chemistry
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drug effects
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Chronic Disease
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Drugs, Chinese Herbal
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pharmacology
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Male
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Medicine, Chinese Traditional
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Neurotransmitter Agents
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analysis
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Norepinephrine
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analysis
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Rats
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Rats, Sprague-Dawley
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Serotonin
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analysis
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Stress, Psychological
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metabolism
9.Toxic effect of oral ricin on the mouse intestinal tract and immune organs
Linna LIU ; Hongwei GAO ; Ying DONG ; Zhiping XIA ; Xiaohuan ZOU ; Jiping LI ; Wensen LIU ; Jiayu WAN
Chinese Journal of Veterinary Science 2009;29(7):898-900
The experiment aimed to study the toxic effect of oral ricin on gastrointestinal tract and immune organs of mice with the dose of 1/5 LD50.In early days of intoxication,there was an obviously decrease in daffy weight and relative weight of thymus and spleen,fllowing the excretion of toxin,they had a trend of recovering to the normal state.Also,results of pathological section,scanning electron microscope and transmission electron microscope showed that ricin would induce a series of pathological reaction in intestines,meanwhile,the splenocytes displayed significant symptom of apoptosis and necrosis.
10.Dynamic Effects of Prolonged Inhaling High Concentration of Oxygen on Collagen Type Ⅰand Ⅳ Massenger Ribonucleic Acid Expression in the Lung Tissue of Neonatal Rats
xue-yan, LIU ; qun-qing, LIU ; wan-yi, WANG ; xin-dong, XUE
Journal of Applied Clinical Pediatrics 1986;0(02):-
Objective To explore the dynamic changes of collagen type Ⅰand Ⅳ messenger ribonucleic acid(mRNA)expression in the lung tissue of neonatal rats after inhaling high concentration of oxygen and the role of collagen type Ⅰand Ⅳ mRNA in chronic lung disease(CLD)induced by hyperoxia.Methods Full-term newborn rats were grouped according to inhale the concentration of oxygen into hypero-xia group and air control group after birth within 12 hours.Lung histological section at day 1,3,7,14 and 21 in 2 groups were prepared for hematoxylin-eosin staining and the detection of mRNA level of collagen type Ⅰand Ⅳ by in situ hybridization.The results were analyzed with SPSS 11.0 software.Results Compared with air control group,inflammation response was seen in early stage,the arrest of lung development was evident after 7 d of oxygen exposure,at last interstitial fibrosis.It was shown that the positive expression of collagen typeⅠ was mainly in the alveolar epithelial cells and endothelial cells by in situ hybridization.The expression of collagen typeⅠ mRNA was weakened compared to air group on 7 d(P0.05).Conclusions Prolonged hyperoxia may cause the onset of arrested lung development and lung fibrosis,which are similar to the changes of chronic lung disease.The collagen type Ⅰand Ⅳ mRNA expressions are not parallel to their protein contents,suggesting the main modulation of these collagens may be not at transcriptional level.