3.Mechanism of tumor necrosis factor-like weak inducer of apoptosis on the synthesis of Matrix meta-lloproteinase-1 in fibroblast-like synoviocytes of rheumatoid arthritis
Liping XIA ; Hui SHEN ; Jing LU ; Weiguo XIAO
Chinese Journal of Rheumatology 2009;13(9):627-629
Objective To study whether p38 mitogen-activated protein kinases (p38MAPK) signal pathway were activated in the process of tumor necrosis factor-like weak inducer of apoptosis (TWEAK) which then induces rheumatoid arthritis (RA) fibroblast-like synoviocyte (FLS) to synthesize matrix metalloproteinase-1 (MMP-1) and look for the relative mechanisms of how TWEAK was involved in the destruction of articular bones and cartilage. Methods RA FLS were primarily cultured and stimulated with TWEAK. FLS were pretreated with SB203580 or not. ELISA was used to detect the concentration of MMP-1 in cell-cultured supernatant. Western blotting was used to detect the expression of p-p38MAPK and P65 in RA FLS. Results TWEAK (100 ng/ml) could induce RA FIS to synthesize MMP-1. SB203580 could partially inhibite the expression of MMP-1 producted by RA FLS which was induced by TWEAK. TWEAK could make p38MAPK phosphorylated and increase the expression of P65 protein in the cell nucleus. Conclusion TWEAK induces RA FLS to synthesize MMP-1. In this process, p38MAPK signal transduction pathway is activated and then induce the expression of NF-κB.
4.Clinical experiences of laparoscopic radical nephrectomy in 600 cases
Gangyue HAO ; Jing XIAO ; Hongliang SHEN ; Peiqian YANG
International Journal of Surgery 2012;39(10):665-668
Objective To report the author' s experience with the laparoscopic radical nephrectomy and eraluate it's saftey and therapy.Methods The retrospective analysis was performed on 600 consecutive patients undergoing laparoscopic radieal nephrectomy in the author' s institute from March 2004 to March 2012.There were 380 male and 220 female patients with the average age of 55 years (16-84 years).Five hundred and sixty cases and 40 cases underwent retroperitoneal and transperitoneal surgery respectively.The operative time,estimated blood loss,postoperative hospital stay,complications and the time of complete convalesceuce were recorded.Results Thirteen cases were converted to open surgery due to severe adhesion and severe bleeding.The mean operative time was 110 min(70-320 min).The medium estimated blood loss was 90 mL(30-830 mL) and 15 cases required blood transfusion.The mean drainage time was 50 h(36-72 h),the mean time to first oral intake was 23 h (19-43 h) and the mean postoperative hospital stay was 7 d(5.5-10 d).There were 4 patients with severe complication.Conclusion Laparscopic radical nephrectomy is safe and effective.It has been the standard therapeutic modality for localized renal cell carcinoma.
5.Pusher curving technique for preventing tilt of femoral Günther Tulip inferior vena cava filter: in vitro study
Liang XIAO ; Jing SHEN ; Desheng HUANG ; Ke XU
Chinese Journal of Radiology 2011;45(4):383-387
Objective To determine whether the adjustment of the pusher of GTF was useful to decrease the degree of tilting of the femoral Ginther Tulip filter (GTF) in an in vitro caval model. Methods The caval model was constructed by placement of a 25 mm × 100 mm and two 10 mm ×200 mm Dacron graft inside a transparent bifurcate glass tube. The study consisted of two groups: left straight group (GLS) (n =100) and left curved group (GLC) (n = 100). In the GLC, a 10° to 20° angle was curved on the introducer.The distance (DCH) between the caval right wall and the hook was measured. The degree of tilting (DT) was classified into 5 grades and recorded. Before and after the GTF being released, the angle (ACM1, 2)between the axis of IVC and the metal mount, the distance (DCM1) between the caval right wall and the metal mount, the angle (ACF) between the axis of IVC and the axis of the filter and the diameter of IVC (DIVC) were measured. The data were analyzed with Chi-Square test, t test, rank sum test and Pearson correlation test. Results The degree of GTF tilting in each group revealed a divergent tendency. In group LC, the apex of the filter tended to be grade Ⅲ compared in group LS (x2 value 37. 491 ,P <0.01).The differences of most variables between GLS and GLC were considered as statistical significance(16. 60° vs.3.05°, 20.60° vs. 3.50°, -3.90°vs. -0.40°, 2.98 mm vs. 10.40 mm, - 10.95° vs. -0.485°,13. 17 mm vs. 10.06 mm, - 1.70° vs. 0.70°, t or Z values - 12. 187, - 12. 188, -8.545, -51.834,-11. 395,9. 562, -3. 596, P < 0. 01). There exist significant positive correlation between ACM1 and ACF,ACM1 - ACM2 and DCH1 - DCH2 in each group, respectively(r values 0. 978,0. 344,0. 879,0. 627 ,P <0. 01),while significant negative associations are detected between DCH1 and ACF in each group , ACP and ACF in group LC(r values -0.974, -0.322, -0.702,P<0.01). Conclusion The technique of adjusting the orientation of filter pusher had minimized the incidence and extent of GTF filter tilting in vitro.
6.The heart functions of preschool and school-age children by means of electrical impedance tomography.
Jian-Xin SHEN ; San-Ming ZHANG ; Xiao-Jing WEN
Chinese Journal of Applied Physiology 2008;24(1):17-124
Age Factors
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Cardiac Output
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physiology
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Cardiography, Impedance
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methods
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Child
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Child, Preschool
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Electric Impedance
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Female
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Heart
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physiology
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Humans
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Male
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Stroke Volume
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physiology
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Tomography
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methods
7.Expression of nuclear factor kappa B in the spinal ganglions of adjuvant arthritis rats
Hui SHEN ; Jing LU ; Weiguo XIAO ; Xiubi FANG
Chinese Journal of Pathophysiology 1986;0(03):-
AIM: To study the expression of nuclear factor kappa B (NF-?B) in the L 1-L 4 spinal ganglions of adjuvant arthritis (AA) rats, to explore the neuroendocrinoimmunological mechanism of rheumatoid arthritis (RA). METHODS: The expression of the NF-?B protein was detected by immunohistochemistry method and the alteration of the active NF-?B protein (nuclear protein) was investigated by Western blotting analysis. RESULTS: The expression of NF-?B protein in L 1-L 4 spinal ganglions of AA rats was much higher than that in the healthy controls ( P
8.Regulation mechanism of autophagy-related protein LC3 by c-Jun in methotrexate resistant human choriocarcinoma JEG-3 cells
Yun SHEN ; Yang XIANG ; Changji XIAO ; Jing ZHAO
Chinese Journal of Obstetrics and Gynecology 2014;(7):531-535
Objective To explore the regulation mechanism of autophagy-related protein, microtubule-associated protein 1 light chain 3 (LC3), via c-Jun in methotrexate resistant human choriocarcinoma JEG-3 cell lines. Methods Human choriocarcinoma JEG-3 cell lines, and methotrexate resistant choriocarcinoma JEG-3 (JEG-3/MTXR) cell lines were used in our present study. Phosphorylation c-Jun (p-c-Jun) was evaluated after exposure to 0.02 ng/ml methotrexate for 72 hours in both cells by western blot. c-Jun gene was knockdown by small interference RNA (siRNA) in JEG-3/MTXR cells, and LC3 was evaluated by western blot and reverse transcription-PCR. The binding of LC3 promoter with c-Jun protein was detected via chromatin immunoprecipitation assay (ChIP) with or without 0.02 ng/ml methotrexate exposure. Results The results showed that p-c-Jun was up-regulated after methotrexate treatment for 72 hours (1.99±0.20, versus 0.20±0.06 at 0 hour;P<0.05) by western blot analysis in JEG-3/MTXR cell lines. Further investigation demonstrated that c-Jun-siRNA could inhibit the up-regulation of LC3 formation and after methotrexate exposure (LC3 mRNA:1.24±0.17 versus 3.03±0.43;LC3 protein:0.52±0.07 verus 1.20± 0.15; all P<0.05). The binding of LC3 promoter by c-Jun protein was up-regulated after methotrexate treatment by the method of ChIP in methotrexate resistant JEG-3/MTXR cells [(2.95 ± 0.35) times]. Conclusion Autophagy-related gene LC3 expression regulated by c-Jun protein may be involved in the effect mechanism of the development of methotrexate resistance in choriocarcinoma JEG-3 cells.
9.Analysis on medication principles for cough based on experience of Xu Di-hua, descendant of Meng He Medical School.
Xiao-jing XU ; Li-min XU ; Chun-feng SHEN ; Cai-hua WANG ; Chun-ti SHEN
China Journal of Chinese Materia Medica 2015;40(21):4301-4305
Based on the software of traditional Chinese medicine inheritance support system (TCMISS), this article aims to analyze the experience and composition rules for cough from the descendant of Meng He Medical School, Xu Di-hua. The cough cases treated by Xu Di-hua were collected, and recorded into TCMISS (V2.0). Data mining methods such as Apriori algorithm and complex system entropy cluster were used to analyze the medication principles of Xu Di-hua for cough from pathogenesis and therapeutie aspects, and dig out the frequency of the herbs in prescription, core medicine and new combinations. The experience of curing cough from Professor Xu Di-hua were well found in the research. He is good at choosing prescriptions accurately, and pays attention to simultaneous use of cold and moisture drugs with combination of tonification and purgation. He is skilled in adding or reducing materia medica flexibly, as well as regulating lung to relieve cough and eliminating phlegm by clearing heat.
Algorithms
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Cough
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drug therapy
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Data Mining
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Drug Therapy, Combination
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Drugs, Chinese Herbal
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chemistry
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therapeutic use
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Female
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Humans
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Male
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Materia Medica
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Medicine, Chinese Traditional
10.Preparation and identification of scFv and bsFv against transferrin receptor.
Jing, LIU ; Daiwen, XIAO ; Xiaoou, ZHOU ; Xue, WEN ; Hong, DAI ; Zhihua, WANG ; Xin, SHEN ; Wei, DAI ; Daofeng, YANG ; Guanxin, SHEN
Journal of Huazhong University of Science and Technology (Medical Sciences) 2008;28(6):621-5
To obtain single chain variable fragment (scFv) and bivalent single chain variable fragment (bsFv) against transferrin receptor, up-stream and down-stream primers were designed according to the complementary sequences of FR1 region of variable heavy (VH) and FR4 of variable light (VL), respectively, which contained inter-linker G4S and the restriction endonuclease SfiI, AscI and NotI. Two pieces of scFv fragments were first amplified through PCR and then inserted into plasmid pAB1, which could express scFv protein once induced by IPTG in the host bacteria. To express scFv and bsFv, E. coli TG1 was cultured in LB broth and was induced by IPTG. The restriction enzyme digestion map and DNA sequencing demonstrated that scFv and bsFv genes were successfully inserted into the expression plasmid. SDS-PAGE and Western blotting revealed the protein band at 35kD and 60kD, which were consistent with the molecular weight of scFv and bsFv respectively. Flow cytometry showed that scFv and bsFv harbored the specific binding activity with TfR expressed in various tumor cells, and the avidity of bsFv was higher than that of the parent scFv.
Base Sequence
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Cloning, Molecular
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Escherichia coli/genetics
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Escherichia coli/metabolism
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Genetic Vectors/genetics
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Hep G2 Cells
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K562 Cells
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Molecular Sequence Data
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Receptors, Transferrin/*immunology
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Recombinant Fusion Proteins/biosynthesis
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Recombinant Fusion Proteins/genetics
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Single-Chain Antibodies/*biosynthesis
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Single-Chain Antibodies/genetics