2.Effect of β-adrenergic receptor stimulation on vascular smooth muscle cell apoptosis
Jing MA ; Ming ZHENG ; Shi-wen WANG ; Yusheng ZHAO
Chinese Journal of Rehabilitation Theory and Practice 2004;10(5):271-273
ObjectiveTo investigate the effect of β-adrenergic receptor(β2AR) stimulation on vascular smooth muscle cell apoptosis in physiological state and receptor overexpression model.Methodsβ2AR overexpression model was established by transgenic techniques. Hoechst 33342 staining as well as flow cytometer(FCM) detected were chosen to measure the incidence of vascular smooth muscle cell apoptosis.ResultsVascular smooth muscle cell exhibited significantly fewer viable cell rate when stimulated with β2AR agonist isoproteronol for 48 hours compared with control (P<0.01),while no difference at the time point of 24 hours. Much fewer viable rate detected by FCM and high apoptotic rate by Hoechst staining were observed when VSMCs overexpressing β2AR were stimulated with isoproteronol for 24 hours (P<0.01).ConclusionStimulation of physiological and overexpressing β2AR could induce apoptosis of vascular smooth muscle cell.
3.Preliminary Research on Case Teaching Method for Pharmacy Administration Science
Xiaopeng SHI ; Shanbo MA ; Yin WU ; Jinyi CAO ; Aidong WEN
China Pharmacist 2015;18(10):1834-1836
To explore and discuss the application of case teaching method for pharmaceutical administration science according to the actual teaching situation and the teaching experience of the authors. The teaching effects can be improved by the method, which is worthy of promotion and popularization.
4.Effects of RNA Interfering of MBP-1 on Proliferation of Saos-2 Cell Line
Xinhe SHI ; Zhe GENG ; Xingchen SHI ; Kejun MA ; Hongwen ZHU ; Wen REN ; Yali ZHOU
Journal of China Medical University 2016;45(7):604-609
Objective To investigate the effects of c?myc promoter binding protein 1(MBP?1)gene on the proliferation of human Saos?2 osteo?sarcoma cells in vitro. Methods Saos?2 cells were divided into three groups:blank control group(untransfected cells),negative group(cells transfected with missense sequence)and experimental group(cells transfected with MBP?1 shRNA). Two MBP?1 shRNA sequences and one neg?ative control shRNA sequence were designed ,synthesized and cloned into pSIREN?retroQ plasma. Then the recombinant plasmids were construct?ed and transfected into human Saos?2 osteosarcoma cells by Lipofectamine 2000. The expressions of MBP?1 mRNA and protein in Saos?2 cells were detected by real?time PCR and Western blot ,respectively. The effects of altered expression of MBP?1 on cell proliferation were measured by CCK?8 cell proliferation assay. The expressions of cyclin D1 and cyclin E in Saos?2 were determined by Western blot. Results PCR and sequenc?ing results indicated that the recombinant plasmids pSIREN?retroQ was constructed. The relative expression level of MBP?1 mRNA in the MBP?1 siRNA transfection group was significantly decreased than that in blank control group(P<0.05). Compared with the blank control group,the ex?pression levels of MBP?1 protein in the experimental group also significantly decreased. The proliferation abilities of Saos?2 cells at 48,72,and 96 hours after MBP?1 siRNA transfection were significantly increased than those in the blank control group(P<0.05). Compared with the blank con?trol group,the expression levels of cyclin D1 and cyclin E protein in the experimental group also significantly increased(P<0.05). Conclusion Knockdown of the expression of MBP?1 gene promotes the proliferation of human Saos?2 osteosarcoma cells. MBP?1 gene may become the new tar?get of gene therapy for osteosarcoma.
5.Construction of shSet7/9 vector and its function in HepG2
Kejun MA ; Xingchen SHI ; Ping LI ; Xiaoqiang LI ; Wen REN ; Long QIN ; Xinhe SHI
Journal of Xi'an Jiaotong University(Medical Sciences) 2015;(6):753-757
Objective To silence human gene Set7/9 and screen out stable transfection cell line in hepatocellular carcinoma cell line HepG2 so as to investigate the impact of down-regulation of Set7/9 in cell line HepG2 and provide experimental foundation for studies on the effect of set7/9 in HepG2.Methods The target oligo was designed and synthesized;shRNA interference vector and the control vector were constructed and transfected into HepG2 cells;the stable transfection cells were screened out.Then Real-time PCR and Western blot were performed to detect the silence of Set7/9 according to both gene expression and protein expression level. Results The shRNA interference vector was constructed and transfected into HepG2 cells successfully.Compared with that in the negative control group,the expression of Set7/9 was dramatically downregulated (P < 0.05 ). Meanwhile,the expression of related protein Sirt1 and Suv39h1 was upregulated 8.4 folds and 1.1 fold, respectively.Conclusion Downregulation of Set7/9 expression can upregulate Sirt1 and Suv39h1,suggesting that Set7/9 may affect the activity of HepG2 cell lines.
6.A Randomized Controlled Trial on Qi-tonifying, Stasis-resolving and Kidney-tonifying Treatment of Lumbar Disc Herniation
Jinhai XU ; Jing WANG ; Jie YE ; Junming MA ; Xuequn WU ; Wen MO ; Qi SHI
World Science and Technology-Modernization of Traditional Chinese Medicine 2013;(5):1074-1080
This study was aimed to evaluate the clinical effect and safety of Chinese medicine treatment of lumbar disc herniation with qi-tonifying, stasis-resolving and kidney-tonifying method. The randomized con-trolled trial (RCT) was applied in the study to evaluate the clinical effect of qi-tonifying, stasis-resolving and kidney-tonifying method in the treatment of lumbar disc herniation . A total of 122 lumbar disc herniation pa-tients were randomly divided into the treatment group ( n = 61 ) and the control group ( n = 61 ) . Chinese medicine treatment with the qi-tonifying, stasis-resolving and kidney-tonifying method was applied in the treatment group . And Celecoxib and Methycobal were orally administered in the control group . Then , the VAS scores, JOA scores, Oswestry disability index (ODI) were recorded and analyzed pre-treatment, four weeks af-ter treatment and the twelfth week of follow-up in order to evaluate the clinical effect . Adverse reactions were also observed and recorded at the same time to give a comprehensive evaluation on its safety . The results showed that there were no significant differences between the treatment group and control group in the baseline data before treatment . Hence , data from two groups were comparable . Compared with pre-treatment , the VAS scores and ODI scores were obviously reduced in both groups after four-week treatment . The JOA scores were increased obviously ( P < 0 . 05 ) . There were no statistical differences on ODI scores and JOA scores between two groups . The VAS scores of the treatment group were obviously higher than the control group ( P < 0 . 05 ) . In the twelfth week of follow-up , the VAS scores , ODI scores and JOA scores had increasing tendency in
both groups . There were no statistical differences between two groups . There were no statistical differences on the total effective rate between two groups . In the treatment group , four patients received surgery , four cases lost to follow-up , and four cases with mild adverse event . In the control group , six patients received surgery , three cases lost to follow-up , and two cases with mild adverse event . It was concluded that the RCT of Chi-nese medicine treatment of lumbar disc herniation with q i-tonifying , stasis-resolving and kidney-tonifying method received same clinical effect as the combination of Celecoxib and Methycobal . The Chinese medicine treatment can effectively relieve pain degree of lumbar disc herniation , improve function of the lumbar vertebrae and improve the daily life and social activity ability of patients. The short-term follow-up effects were con-firmed . However , the long-term efficacy still requires further study .
7.Chemical constituents from Callicarpa nudiflora and their cytotoxic activities.
Yan-Chun MA ; Min ZHANG ; Wen-Tong XU ; Shi-Xiu FENG ; Ming LEI ; Bo YI
China Journal of Chinese Materia Medica 2014;39(16):3094-3101
The chemical consitituents from cytotoxic fraction of the Callicarpa nudiflora extract were isolated and purified by a combination of HP-20 macroporous resin, silica gel and Sephadex LH-20 column chromatographies. The structures were elucidated on the basis of the spectroscopic data and comparison of their spectroscopic data with reported data. The cytotoxicity was evaluated by the MTT assay. The 50% and 70% EtOH elutions of EtOH-extract showed significant cytotoxic activities, leading to the isolation of twelve compounds, which were identified as luteoloside(1), lutedin-4'-O-β-D-glucoside(2), 6-hydroxyluteolin-7-O-β-glucoside(3), lutedin-7-O-neohesperidoside(4), rhoifolin (5), luteolin-7, 4'-di-O-glucoside (6), forsythoside B (7), acteoside (8), alyssonoside (9), catalpol(10), nudifloside(11), and leonuride(12). Compounds 3-6, 10 and 12 were isolated from this genus for the first time, and compound 9 was isolated from this plant for the first time. The cytotoxicity assay demonstrated that flavonoids 1-6, in various concentrations, showed monolithic proliferation inhibitory activities against Hela, A549 and MCF-7 cell lines. Compounds 3, 5 and iridoid glycoside 11 possessed higher cytotoxicacivities. In short, flavonoids are the main components of cytotoxic extract from C. nudiflora, while phenylethanoid glycosides are the predominant ingredient but inactive to cancer cell lines. In addition, the minor iridoid glycoside expressed weak cytotoxic activity.
Callicarpa
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chemistry
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Cell Proliferation
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drug effects
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Cytotoxins
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chemistry
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isolation & purification
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pharmacology
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Drugs, Chinese Herbal
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chemistry
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isolation & purification
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pharmacology
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Humans
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MCF-7 Cells
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Molecular Structure
8.Effect of Yangxue Qingnao Granule on the Expression of CD11b in CA1 Region of Hippocampus of Vascular Dementia Rats.
Jing LI ; Yuan-yuan MA ; Bin LIU ; Wen-jing MAO ; Jin-xia ZHANG ; Shi-ying LI
Chinese Journal of Integrated Traditional and Western Medicine 2016;36(5):619-623
OBJECTIVETo observe the effect of Yangxue Qingnao Granule (YQG) on the expression of CD11b in CA1 region of hippocampus of vascular dementia rats, and to explore its regulation on microglias.
METHODSTotally 144 SD rats were randomly divided into the sham-operation group, the vascular dementia model group (model), and the YQG treated group (treated). The vascular dementia rat model was prepared by modified Pulsinelli's four-vessel occlusion. Rats in the sham-operation group and the model group were administered with normal saline -(at the daily dose of 10 mL/kg) by gastrogavage, while those in the treated group were administered with YQG (0.32 g/mL, at the daily dose of 10 mL/kg) by gastrogavage. All administration was performed once per day for 8 successive weeks. The expression of CD11b in CA1 region of hippocampus of vascular dementia rats was detected at week 1, 2, 4, and 8, respectively.
RESULTSCompared with the sham-operation group, the expression of CD11b in CA1 region of hippocampus of vascular dementia rats were significantly enhanced in the model group at each time point (P < 0.01). Compared with the model group, the expression of CD11b in CA1 region of hippocampus of vascular dementia rats significantly decreased in the treated group at each time point (P < 0.01), especially at week 2.
CONCLUSIONObvious activation and proliferation of microglias could be seen in CA1 region of hippocampus of vascular dementia rats, and YQG could inhibit activation and proliferation of microglias.
Animals ; CA1 Region, Hippocampal ; drug effects ; metabolism ; CD11b Antigen ; metabolism ; Dementia, Vascular ; drug therapy ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; Microglia ; drug effects ; Random Allocation ; Rats ; Rats, Sprague-Dawley
9.Sensing scheme of minim liquid density based on Lamb-wave.
Tao HAN ; Wen-kang SHI ; Wei-fang MA ; Yi HOU
Chinese Journal of Medical Instrumentation 2002;26(3):172-209
This paper presents and discusses the liquid density sensing potential of a delay line Lamb-wave sensor configuration, where Lamb wave is excited only by interdigital transducer (IDT), without membrane when fabricated. An optimized cut orientation and the normalized thickness of substrate by using genetic algorithm is presented to improve the sensitivity of the sensor and to get low noise. Experimental measurements performed on Lamb-wave device fabricated from Y-Z LiNbO3 in a delay line configuration are found to be in agreement with theoretical calculations.
Algorithms
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Biosensing Techniques
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instrumentation
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methods
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Equipment Design
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Models, Theoretical
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Sensitivity and Specificity
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Transducers
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Ultrasonics
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Water
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analysis
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chemistry
10.Icariin promote maturation of osteoblasts in vitro by an estrogen-independent mechanism.
Wen-Gui SHI ; Xiao-Ni MA ; Yan-Fang XIE ; Jian ZHOU ; Jian ZHOU
China Journal of Chinese Materia Medica 2014;39(14):2704-2709
OBJECTIVETo investigate the estrogenic activity of icariin and genistein with estrogen-dependent human breast cancer (MCF-7) cells.
METHODMCF-7 cells were incubated with media containing 5% charcoal dextran-treated FBS in phenol red-free media for 48 h. CCK-8 kit was used to study the impact of defferent concentration of icariin and genistein on MCF-7 proliferation in vitro. Optimal concentration icariin and genistein were added into medium and total RNA was isolated after 12, 24, 36, 48 h. The gene expression of ERalpha, ERbeta, PS2, and PR were investigated by Real-time RT-PCR Total protein was also isolated and secretion of ERalpha, ERbeta, PS2, and PR were examined by Western blot.
RESULT10 micromol x L(-1) icariin and genistein could promote the proliferation of MCF-7 evidently. However, the ability of genistein to promote the proliferation was better than icariin. With the concentration of 10 micromol x L(-1), genistein group had a stronger expression of ERa, PS2 and PR mRNA levels than icariin while ERbetaexpression had no significant difference in two group. The same effects were detected by western blotting.
CONCLUSIONBoth genistein and icariin have a strong estrogen-like effect, but the estrogenic activity of genistein is stronger than icariin. It showed that the activity of icariin is stron-ger than genistein to promote ROB maturation. So it must be that icariin promotes the maturation of osteoblasts in vitro by a estogen-independent mechanism.
Cell Proliferation ; drug effects ; Estrogen Receptor alpha ; genetics ; metabolism ; Estrogen Receptor beta ; genetics ; metabolism ; Estrogens ; pharmacology ; Flavonoids ; pharmacology ; Gene Expression Regulation ; drug effects ; Genistein ; pharmacology ; Humans ; MCF-7 Cells ; Osteoblasts ; cytology ; drug effects ; metabolism ; Presenilin-2 ; metabolism