1.Construction of eukaryotic expression bicistron plasmid vector pcDNA3.1-hOPG and its expression in vitro
Hua TANG ; Ping GONG ; Zhen TAN ; Dapeng LIAO
Journal of Third Military Medical University 2003;0(23):-
Objective To construct eukaryotic expression bicistron plasmid vector pcDNA3.1-hOPG,and to detect the expression of the plasmid in vitro.Methods hOPG cDNA fragment was extracted from plasmid MGC:29565,amplified by PCR,and inserted into pcDNA3.1(-)vector,then identified by restriction endonuclease digestion and nucleotide sequencing.C2C12 cells were transfected with the plasmid using Lipofectamine 2000.The expression of OPG protein was detected by immunohistochemistry techniques and bone wafer pit assay.Results The constructed recombinant plasmid contained the sequence of hOPG gene.After transfection with the plasmid,active OPG protein could be expressed in C2C12 cells.Conclusion The constructed eukaryotic expression bicistron plasmid vector pcDNA3.1-hOPG can express OPG in vitro,which is the basis for further study on the treatment of bone absorption caused by abnormal activity of osteoclasts.
2.Effects of inhibiting Kit signal pathway on proliferation and spontaneous rhythmic motility of interstitial cells in bladder of guinea pigs
Shiming TANG ; Dapeng HE ; Zhong WANG ; Fuxian GU ; Mingrong WANG
Journal of Third Military Medical University 2003;0(23):-
Objective To investigate the role of Kit signal pathway in the proliferation and spontaneous rhythmic motility of interstitial cells(ICs)in the bladder of guinea pigs.Methods Fourteen Guinea pigs aged 45-60 days were administered with Imatinib intragastrically for 20 and 40 days.Three were fed on normal saline as control.The morph and number of ICs were observed by immunofluorescence,Kit protein expression was detected by Western blot and spontaneous rhythmic motility of bladder were investigated in in vitro organ bath.Results Quite a few ICs were observed in the bladder of controls.After administration of Imatinib,the amount of ICs and the expression of Kit protein decreased gradually,and the spontaneous rhythmic contraction weakened.Conclusion ICs might be the pacemaker of bladder in guinea pigs.Kit protein might play an important role in the survival and functional maintenance of ICs.
3.Effect of morphine on serum hungry-induced apoptosis in cardiac myocytes
Bo TANG ; Hongxin WANG ; Dapeng WANG ; Xiaochun YU
Chinese Pharmacological Bulletin 2003;0(08):-
Aim Myocardial cells of neonatal rats were cultured in vitro to study aim of morphine on serum hungry-induced apoptosis in cardiac myocytes and its mechanism.Methods Myocardial cells of neonatalrats were cultured in vitro.48 hours later,different agents were added to cardiac myocytes.The cellular survival was determined with MTT colormeteric assay;apoptosis rates were determined by Annexin V-FITC/PI;cell cycle was determined by flow cytometry;Caspase-3 and PKC were investigated by Western blot.Results Free-serum induced apoptosis in cardiac myocytes was shown after 48 hours;morphine(1 ?mol?L-1)could inhibit apoptosis in cardiac myocytes,manifestation,apoptosis rates were decreased,Caspase-3 experssion was decaeased Naloxone at 10 ?mol?L-1 inhibited the promoting effects of morphine;GF109203X at 10 ?mol?L-1 or Staurosporine at 1 ?mol?L-1 could inhibit the promoting effects of morphine,manifestation,apoptosis rates were increased,Caspase-3 experssion was incaeased,PKC expression was decreased.Conclusion morphine inhibited serum hungry-induced apoptosis in cardiac myocytes via PKC signal transduction pathway.
4.?-Opioid receptor inhibits serum deprivation-induced apoptosis of neonatal cardiac myocytes through activation of protein kinase C signal transduction pathway
Dapeng WANG ; Hongxin WANG ; Bo TANG ; Xiaochun YU
Chinese Pharmacological Bulletin 1987;0(01):-
Aim To investigate the possible mechanism of ?-opioid receptor inhibiting serum deprivation-induced apoptosis of neonatal cardiac myocytes.Methods Myocardial cells of neonatal rats were cultured in vitro,after 48 h,the medium was changed to serum-free DMEM.The experimental groups were:A:Normal;B: Model;C: Model+DADLE(0.1 ?mol?L-1);D:Model+DADLE(0.1 ?mol?L-1)+naltrindole(10 ?mol?L-1);E:Model+DADLE(0.1 ?mol?L-1)+GF109203X(10 ?mol?L-1);F:Model+DADLE(0.1 ?mol?L-1)+staurosporine(1 ?mol?L-1).The cell viability was determined with MTT colormeteric assay;apoptosis index and the percentage of G0 in cell cycle were determined by flow cytometry;Caspase-3 and PKC signal pathway was investigated by Western blot.Results ?-opioid receptor agonist [D-Ala2,D-Leu5]-enkephalin(DADLE) could significantly inhibit serum deprivationinduced apoptosis of neonatal cardiac myocytes,which increased the survival index of cardiac myocyte,percentage of G2/M in cell cycle and the expression of PKC,decreased the apoptotic index of cardiac myocyte,percentage of G0/G1 in cell cycle and the expression of activate caspase-3.The protective effect of DADLE was obviously blocked by ?-opioid receptor antagonist Naltrindole at 10 ?mol?L-1,the PKC inhibitor GF109203X at 10 ?mol?L-1 and staurosporine at 1 ?mol?L-1,which decreased the survival index of cardiac myocyte,percentage of G2/M in cell cycle and the expression of PKC,increased the apoptotic index of cardiac myocyte,percentage of G0/G1 in cell cycle and the expression of caspase-3.Conclusion These findings suggested that ?-opioid receptor activation might be a potential survival factor against serum deprivation-induced myocardial cell apoptosis and this cardioprotective effect might be via PKC pathway.
5.Imaging diagnosis of pulmonary lymphangitic carcinomatosis
Zhidan LEI ; Wulin JIA ; Dapeng SHI ; Xitao MA ; Xueyi TANG ; Zhigang YANG
Chinese Journal of Postgraduates of Medicine 2006;0(25):-
Objective To study the imaging findings of pulmonary lymphangitic carcinomatosis (PLC) and discuss it′s clinical value. Methods The imaging materials of 40 cases with PLC which were proved by bronchoscopic or pleural or open-lung biopsy were retrospectively analyzed, and the clinical application of imaging diagnosis were investigated. Results The primary tumorous pathological types of PLC included:13 cases peripheral type carcinoma of lung,2 cases central type carcinoma of lung,11 cases mammary cancer,6 cases gastric carcinoma,4 cases pancreatic carcinoma,3 cases renal carcinoma,1 case colon carcinoma. The major imaging features of PLC were showed as pleural nodes, lobular core nodes, intralobular small reticular and nodular shadows, small beaded thickened interlobular septums, beaded thickened bronchovascular bundles, tumescent pulmonary hilar and/or mediastinal lymph nodes. Conclusion PLC possess relative imaging feature, particularly HRCT may accurately reflect pathological feature of PLC. Imaging diagnosis is a satisfactory method in diagnosing pulmonary lymphangial metastasis of malignant tumor.
6.Prevalence and risk factors of syphilis in men who have sex with men in 16 districts in China
Yan XIAO ; Chunmei LI ; Fan Lü ; Houlin TANG ; Canrui JIN ; Dapeng ZHANG ; Liyan WANG ; Yujiang JIA ; Jiangping SUN
Chinese Journal of Dermatology 2008;41(6):353-356
Objective To examine the prevalence of and risk factors for syphilis among men who have sex with men (MSM) in 16 cities/districts, 6 provinces in China. Methods Based on a previous survey on the entertainment place and population size of MSM, a cross-sectional study was conducted in 2007 using non-probability sampling method among MSM in 16 cities/districts in China.Information about the demography,risk behavior and syphilis infection was collected through questionnaires from MSM. Univariate and multivariate stepwise logistic regression model was constructed to evaluate independent factors associated with syphilis. Results A total of 4211 participants were recruited. The mean age of these participants was 28 years. Among them, 66.7% were self identified as homosexual, 72.5% were unmarried, and 76.4% had an education beyond senior middle school; in the past 6 months, 8.7% had sexual contact with more than 10 male sex partners, 5.6% had ever paid for male sex service, 11.3% had provided commercial sex service to males, and 18.9% had sexual contact with females, while condom was consistently used in only 36.3% of male-to-male anal sexual behavior. Syphilis was diagnosed in 10.6% of these participants. The risk factors independently associated with syphilis included being older than 25 years (adjusted OR = 1.7; 95% CI:1.3-2.0), local residency (adjusted OR = 1.3; 95% CI: 1.0-1.7), being recruited from a bar or a park (adjusted OR = 1.3; 95% CI: 1.1-1.7), having ever paid for a male sex service(adjusted OR = 1.5; 95% CI: 1.0-2.1),inconsistent condom use during male-to-male anal sexual behavior(adjusted OR = 1.2; 95% CI: 1.0-1.5 ), self reported sexually transmitted infection history (adjusted OR = 1.3; 95% CI: 1.0-1.7). Conclusions There is a high prevalence of syphilis in MSM in the 16 Chinese cities where male-to-male commercial sex service and unsafe sexual behavior are common. Further efforts are urgently needed to scale-up the prevention of and treatment program for syphilis among MSM, which should include syphilis and HIV screening as well as education on the use of condom.
7.Codon usage bias in the straw mushroom Volvariella volvacea.
Wei JIANG ; Beibei LÜ ; Jianhua HE ; Jinbin WANG ; Xiao WU ; Guogan WU ; Dapeng BAO ; Mingjie CHEN ; Jinsong ZHANG ; Qi TAN ; Xueming TANG
Chinese Journal of Biotechnology 2014;30(9):1424-1435
We analyzed the whole genome coding sequence of Volvariella volvacea to study the pattern utilization of codons by Codon W 1.4.2. As results, 24 optimal codons were identified. Moreover, the frequency of codons usage was calculated by CUSP program. We compared the frequency of codons usage of V. volvacea with other organisms including 6 modal value species (Homo sapiens, Saccharomys cerevisiae, Arabidopsis thalian, Mus musculus, Danio rerio and Drosophila melanogaster) and 4 edible fungi (Coprinopsis cinerea, Agaricus bisporus, Lentinula edodes and Pleurotus ostreatus). We found that there were less differences in 3 edible fungi (excluding Pleurotus ostreatus) than 6 modal value species, comparing with the frequency of codons usage of V. volvacea. With software SPSS16.0, cluster analysis which showed differences in the size of codon bias, reflects the evolutionary relationships between species, which can be used as a reference of evolutionary relationships of species. This was the first time for analysis the codon preference among the whole coding sequences of edible fungi, serving as theoretical basis to apply genetic engineering of V. volvacea.
Agaricales
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genetics
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Animals
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Arabidopsis
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genetics
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Cluster Analysis
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Codon
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DNA, Fungal
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genetics
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Drosophila melanogaster
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genetics
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Humans
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Mice
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Saccharomyces cerevisiae
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genetics
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Software
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Volvariella
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genetics
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Zebrafish
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genetics
8.Ameliorative effects of atractylodin on intestinal inflammation and co-occurring dysmotility in both constipation and diarrhea prominent rats.
Changchun YU ; Yongjian XIONG ; Dapeng CHEN ; Yanli LI ; Bin XU ; Yuan LIN ; Zeyao TANG ; Chunling JIANG ; Li WANG
The Korean Journal of Physiology and Pharmacology 2017;21(1):1-9
Intestinal disorders often co-occur with inflammation and dysmotility. However, drugs which simultaneously improve intestinal inflammation and co-occurring dysmotility are rarely reported. Atractylodin, a widely used herbal medicine, is used to treat digestive disorders. The present study was designed to characterize the effects of atractylodin on amelioration of both jejunal inflammation and the co-occurring dysmotility in both constipation-prominent (CP) and diarrhea-prominent (DP) rats. The results indicated that atractylodin reduced proinflammatory cytokines TNF-α, IL-1β, and IL-6 in the plasma and inhibited the expression of inflammatory mediators iNOS and NF-kappa B in jejunal segments in both CP and DP rats. The results indicated that atractylodin exerted stimulatory effects and inhibitory effects on the contractility of jejunal segments isolated from CP and DP rats respectively, showing a contractile-state-dependent regulation. Atractylodin-induced contractile-state-dependent regulation was also observed by using rat jejunal segments in low and high contractile states respectively (5 pairs of low/high contractile states). Atractylodin up-regulated the decreased phosphorylation of 20 kDa myosin light chain, protein contents of myosin light chain kinase (MLCK), and MLCK mRNA expression in jejunal segments of CP rats and down-regulated those increased parameters in DP rats. Taken together, atractylodin alleviated rat jejunal inflammation and exerted contractile-state-dependent regulation on the contractility of jejunal segments isolated from CP and DP rats respectively, suggesting the potential clinical implication for ameliorating intestinal inflammation and co-occurring dysmotility.
Animals
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Constipation*
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Cytokines
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Diarrhea*
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Herbal Medicine
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Inflammation*
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Interleukin-6
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Myosin Light Chains
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Myosin-Light-Chain Kinase
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NF-kappa B
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Phosphorylation
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Plasma
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Rats*
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RNA, Messenger
9.Three-dimensional constructive interference in steady state sequence for displaying anterolateral ligament of knee joint
Yupeng ZHU ; Xiaoyan TANG ; Lei SUN ; Shaohua WANG ; Feng DUAN ; Chuanyu ZHANG ; Tengbo YU ; Dapeng HAO
Chinese Journal of Medical Imaging Technology 2018;34(5):765-769
Objective To observe the value of three-dimensional constructive interference in steady state (3D-CISS) sequence for displaying anterolateral ligament (ALL) of knee joint.Methods MR scans of right knee joint were performed on 30 healthy volunteers,and the protocol included axial and coronal fat saturation proton density weighted imaging (FS-PDWI) and 3D-CISS sequence.MRP and CPR (reconstructive angles including 0°,30°,60°,90°,120°,150° and 180°) images were generated from of 3D-CISS sequence.The visibility rates of the femoral part,meniscal part,tibial part,meniscal insertion,femoral footprint and tibial footprint of ALL were compared among different protocols.Results The visibility rates of tibial footprint and femoral footprint of ALL on 3D-CISS CPR images were both 96.67% (29/30),and on 3D-CISS MPR images were both 93.33% (28/30),all of them were higher than those on FS-PDWI images (all P<0.017).The visibility rates of tibial part,meniscal part and meniscal insertion of ALL on 3D-CISS CPR images were 96.67% (29/30),83.33% (25/30) and 83.33% (25/30),respectively,and all of them were higher than those on FS-PDWI images (all P<0.05).There was no statistical difference of visibility rate of femoral part between 3D-CISS CPR images and FS-PDWI images (P=0.095).Conclusion 3D-CISS sequence with CPR can significantly enhance the ability to identify ALL.
10.A review of traditional Chinese medicine research in China from 2008 to 2017
Yun XU ; Fang HUANG ; Xing JIANG ; Quiqin WANG ; Huimei WANG ; Weitong LI ; Dapeng TANG
Chinese Journal of Health Management 2019;13(5):441-445
Objective To examine the development of traditional Chinese medicine (TCM) in China and provide reference for the construction of a discipline through a literature review of TCM physique and statistical analyses of papers published between 2008 and 2017. Methods We searched the Chinese journal full?text database for TCM physique research papers, and used bibliometrics to statistically analyze the papers. Results We obtained 2 905 papers from 381 journals, 1 021 institutions, and 6 740 authors. The number of papers increases with the publication time. The cited rate was 87.2%. Each paper was cited 8.74 times. The H?index was 51 and co?authored rate was 83.7%. The cooperation degree was 3.78 with 3 354 words appearing in the keywords of the papers. Conclusion In the past ten years, research on TCM constitution has advanced, strengthening core scientific research. Research quality is high, and the research contents cover a wide range. With the spread of research tentacles, the prospect of further research on this subject is worth looking forward to.