1.Research of survivin antisense oligonucleotide on growth suppression of transplanted human breast cancer in nude mice
Tide DUAN ; Dequan LIU ; Ceyao YANG
Chinese Journal of General Surgery 2001;0(09):-
Objective To explore the suppressive effect of survivin antisense oligonucleotide on transplanted human breast cancer in nude mice. Methods 30 nude mice were transplanted with human breast cancer MCF-7 cells to construct tumor models, which were divided into three groups (10 mice per group): (Lipofectin), ASODN/Lip, and NODN/Lip were injected in to the tumor or around the tumor, respectively. A total of 3 injections were given, once every five days. Observations were made on tumor suppression rate, tumor volume and changes in weight of the animals. TUNEL method and transmission electron microscopy were used to investigate cell apoptosis of the tumor; western blot was used to detemine the expression of survivin protein in the 3 groups. Results The subcutaneous tumor model in nude mice was successfully established. The tumor volume in the group of ASODN/Lip decreased during treatment, the rate of tumor suppression reached 70.10%; while the tumor volume of the other two groups increased. There was no statistical (difference) among the groups in weight changes of the 3 groups mice, the apoptosis rate in ASOND/Lip group was 38.6% detected by TUNEL and the rate was markedly higher than the other two groups(P
2.Morphology of human skin fibroblasts after heat injury in vitro
Dequan LI ; Xiaoyuan HUANG ; Xinghua YANG
Chinese Journal of Tissue Engineering Research 2007;0(11):-
AIM: Previous studies paid more attention to the effect of fibroblast immigration in normal surrounding tissue or blood on wound healing. This study explored the morphological character of fibroblasts after heat injury to support the new operation of autogenous epidermis overlapping denatured dermis. METHODS: The experiment was performed at the State Key Laboratory of Trauma, Burn and Combined Injury in the Third Military Medical University of Chinese PLA from April 2006 to April 2007. The dermal fibroblast from the infant prepuce disposed after surgery was cultured in vitro, and divided into 2 groups. In normal group, the dermal fibroblasts were put in water bath at 37 ℃ for 30 seconds; the cells in experimental group were stimulated at different temperatures (50, 51, 52, and 53 ℃) for different time (30, 60, 90, and 180 seconds). Cell survival was detected by MTT; morphology of injured cells were observed under inverted microscope and transmission electron microscope at 3 hours, 1, 2, 3, 4, 5, and 7 days; cell cycle was detected by flow cytometry. RESULTS: At different temperatures and different time, there were significant differences in the survival rate of fibroblast between the experimental groups and normal group (P
3.Simultaneous Determination of Nine Nucleosides in Fritillaria taipaiensis P. Y. Li from ;Different Producing Areas by RP-HPLC
Jing LUO ; Hua ZHANG ; Dequan ZHANG ; Sisi YANG ; Nong ZHOU
Chinese Journal of Information on Traditional Chinese Medicine 2016;23(9):106-109
Objective To develop a sensitive and reliable RP-HPLC method for the simultaneous determination of nine nucleosides including uracil, cytidine, guanine, uridine, adenine, guanosine, thymidine, adenosine and 2'-deoxyadenosine from Fritillaria taipaiensis P. Y. Li that had been cultivated in different producing areas; To compare the contents of these nucleosides from different producing areas. Methods The analysis was performed on a Venusil MP C18 (2) column (4.6 mm×250 mm, 5 μm) with a gradient of methanol-water at a flow rate of 1.0 mL/min;the detective wavelength was set at 260 nm; the column temperature was set at 35 ℃. Results Uracil, cytidine, guanine, uridine, adenine, guanosine, thymidine, adenosine and 2'-deoxyadenosine were obtained in the good linear range of 0.269 5–16.17 μg/mL, 0.132 1–7.927 5 μg/mL, 0.095 5–5.73 μg/mL, 1.16–69.6 μg/mL, 0.48–28.8 μg/mL, 0.571 5–57.15 μg/mL, 0.526–52.6 μg/mL, 3.307 5–198.45 μg/mL, 0.530 5–31.83 μg/mL, respectively (r≥0.999 5);the recovery was in the range of 96.49%–101.65%(RSD≤2.92%). Conclusion The contents of the nine nucleosides from different producing areas have differences. Fritillaria taipaiensis P. Y. Li from Xianyi Village, Chengkou County, Chongqing City, whether the cultivated ones or wild ones, contain the highest level of nucleosides. The established method can provide references for the perfection of quality standard for Fritillaria taipaiensis P. Y. Li.
4.Expression of the ERG11 gene in fluconazole-resistant Trichosporon asahii
Xiao DING ; Zhikuan XIA ; Dequan ZHANG ; Rongya YANG
Chinese Journal of Dermatology 2017;50(5):355-359
Objective To investigate the role of the ERG11 gene in the drug resistance of Trichosporon asahii (T.asahii), and to explore the relationship between the gene expression and drug concentrations. Methods Stable fluconazole-resistant strains of T.asahii were induced in vitro following exposure to a series of concentrations of fluconazole. Fluconazole-sensitive and-resistant strains of T.asahii were separately cultured in the medium containing fluconazole at concentrations of 0, 0.25, 0.5, 1, 2, 4, 8, 16, 32 and 64 μg/ml. Real-time quantitative PCR was performed to determine the mRNA expression of ERG11 gene. Results In fluconazole-free medium, the fluconazole-resistant strain of T.asahii showed significantly increased mRNA expression of the ERG11 gene compared with the fluconazole-sensitive strain (7.542 ± 5.311 vs. 1.014 ± 0.012, t=3.002, P=0.03). Additionally, the mRNA expression of ERG11 gene was also significantly higher in the fluconazole-resistant strains than the fluconazole-sensitive strains in the culture medium containing fluconazole at different concentrations of 0.25 (9.183 ± 3.226 vs. 3.281 ± 2.068), 0.5(13.657 ± 5.428 vs. 3.459 ± 1.923), 1(15.292 ± 7.007 vs. 3.242 ± 2.530), 2(13.720 ± 8.550 vs. 3.651 ± 0.728), 4(13.949 ± 2.960 vs. 3.969 ± 1.924)and 8(13.123 ± 6.429 vs. 3.824 ± 1.875)μg/ml(all P<0.05). However, no significant correlation was observed between the mRNA expression of ERG11 gene and fluconazole concentrations(fluconazole-resistant strains: rs = 0.229, P = 0.096; fluconazole-sensitive strains:rs=0.166, P=0.357). Conclusion Overexpression of ERG11 gene is associated with fluconazole resistance in T.asahii, but there is no correlation between the mRNA expression of ERG11 gene and fluconazole concentrations.
5.Effect of microevolution on phenotypes and drug resistance of the Trichosporon asahii biofilm
Xin YANG ; Dexiu LANG ; Yong LIAO ; Haitao LI ; Zhuoying PENG ; Junhong AO ; Dequan ZHANG ; Rongya YANG
Chinese Journal of Dermatology 2021;54(1):68-73
Objective:To evaluate the effect of microevolution on phenotypes and drug resistance of the Trichosporon asahii biofilm. Methods:The standard strain of Trichosporon asahii was obtained from the Fungal Biodiversity Institute of the Royal Netherlands Academy of Arts and Sciences, the fluconazole-sensitive primary strain (TO) of Trichosporon asahii was isolated from a case of trichosporonosis diagnosed in the Department of Dermatology, the Seventh Medical Center of Chinese People′s Liberation Army General Hospital in 2000, and the fluconazole-resistant evolved strain (TEVO) of Trichosporon asahii was isolated from the above patient in 2014. Biofilms of the above-mentioned strains were formed in vitro, and tetrazolium salt XTT reduction assay was performed to evaluate growth kinetics of the Trichosporon asahii biofilm, and laser scanning confocal microscopy to determine the thickness of the biofilm; the sessile minimum inhibitory concentrations (SMICs) of fluconazole, itraconazole and voriconazole against the biofilms at different growth stages were determined in vitro for the evaluation of the resistance of the biofilms. One-way analysis of variance was used for comparisons among multiple groups, and Hartley test for testing homogeneity of variance. If the variance was homogeneous, least significant difference test was used for multiple comparisons; if the variance was heterogeneous, Tamhane′ T2 test was used for multiple comparisons. Results:In the adhesion (0 h) and formation stages (4- 24 hours) of the Trichosporon asahii biofilm, the metabolic activity of the evolved strain TEVO was the weakest (adhesion stage: F = 35.705, P < 0.001; formation stage: F = 15.042, P < 0.001) . At 48 hours after adhesion, the biofilms matured, and the TO strain showed the weakest metabolic activity ( F = 10.985, P < 0.001) . In the maturation stage, the biofilm thickness of the TEVO strain (26.1 ± 1.18 μm) was significantly higher than that of the TO strain (22.8 ± 1.73 μm, P = 0.001) , but significantly lower than that of the standard strain (29.5 ± 1.28 μm, P = 0.001) . As drug susceptibility testing showed, the SMICs of azole antifungal agents against the TEVO strain were higher than those against the TO strain in the adhesion and formation stages of the Trichosporon asahii biofilm, and the SMICs of azole antifungal agents against the biofilms of the 3 strains of Trichosporon asahii were all over 1 024 mg/L in the maturation stage of the biofilm. Conclusion:Under the dual pressure of host environment and antifungal drugs, adaptive changes took place in the phenotypes of the Trichosporon asahii biofilm with an increase in the resistance to azole antifungal drugs.
6.Chemical constituents from Morus notabilis and their cytotoxic effect.
Pan ZHEN ; Gang NI ; Xiaoguang CHEN ; Ruoyun CHEN ; Hanze YANG ; Dequan YU
Acta Pharmaceutica Sinica 2015;50(5):579-82
Une new flavonoids named as notabilisin K (1), together with four known compounds, morusin (2), mulberrofuran A (3), neocyclomorusin (4) and mornigrol F (5) are separated from 95% ethanol extracts of the twigs of Morus notabilis. Compounds 2-5 are separated from this plant for the first time. Notabilisin I, notabilisin J exhibits certain effect against cells of HCT-116, HepG2 and A2780 with IC50 values ranging from 1.47 μmol x L(-1) to 5.46 μmol x L(-1). Morusin exhibits strong effect against five kinds of human cancer cells (BGC823, A2780, HCT-116, HepG2 and NCI-H1650) with IC50 values ranging from 0.74 μmol x L(-1) to 1.58 μmol x L(-1).
7.Effect of neoadjuvant chemotherapy on breast cancer cell cycle and breast cancer stem cells
Jiuguang ZHANG ; Dequan LIU ; Ji ZHANG ; Congguo JIN ; Yang LIU ; Yajing WANG ; Yihui ZHAO
Journal of International Oncology 2015;(8):566-568
Objective To investigate the effect of neoadjuvant chemotherapy on the proportion of G0-G1 phase cells and the expressions of ATP-binding cassette sub-family G member 2 (ABCG2)and CD44 +CD24 -/low in breast cancer patients.Methods Sixty untreated cases with breast invasive ductal carcinoma from May 201 3 to March 201 4 were chosen.All patients were tested by core needle biopsy and pathological diagno-sis,then treated by neoadjuvant chemotherapy.The proportion of G0-G1 phase cells and the contents of ABCG2 and CD44 +CD24 -/low before and after therapy were compared.Results After chemotherapy,the contents of ABCG2 and CD44 +CD24 -/low were (25.1 0 ±1 .50)% and (36.40 ±3.80)/1 05 ,and the proportion of G0-G1 phase cells was (70.50 ±1 .50)%,which were higher than those before treatment [(1 5.88 ±1 .22)%, (25.00 ±3.40)/1 05 ,(60.65 ±1 .30)%](t =8.685,P <0.05;t =9.226,P <0.05;t =8.898,P <0.05).All patients completed four courses of chemotherapy,and the cCR rate,cPR rate,SD rate were respec-tively 1 8.3% (1 1 /60),73.3% (44 /60),8.3%(5 /60).Conclusion Cell cycle can be arrested and the proportion of stem cells can be raised after neoadjuvant chemotherapy which has exact curative effect and clini-cal popularization value.
8.The boundary element study on arterial bifurcation vessel flow.
Journal of Biomedical Engineering 2008;25(4):814-817
In this paper a kind of arterial bifurcation vessel's hemodynamic characteristics are studied with the boundary element method, and the blood flowing velocity vector distributions are calculated when there is pathologic change in the branch vessel wall and when there is no pathologic change. The shear stress value at the lateral wall and that at the medial wall are compared when there is pathologic change in the main branch vessel wall and when there is no pathologic change. Moreover, the flow field distribution and the pressure on the particle surface are calculated, when there is flow-round particle at the place of bifurcation. The move tendency of the particle is judged, and the possible factors in causing atherosclerosis are analyzed.
Arteries
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physiology
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physiopathology
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Atherosclerosis
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physiopathology
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Blood Flow Velocity
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Computer Simulation
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Hemodynamics
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physiology
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Humans
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Models, Cardiovascular
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Stress, Mechanical
9.A boundary element analysis on hemodynamic characteristics at the bifurcation of abdominal arterial.
Journal of Biomedical Engineering 2012;29(4):697-700
The hemodynamic characteristics of abdominal arterial bifurcation, such as blood flowing velocity vector, the shear stress at the vessel wall were calculated, studied and compared using the newly-induced boundary element method in this study. It was analysed why the atherosclerosis is asymmetrical at the bifurcation of abdominal arterial. The hemodynamic causes of production and development of the atherosclerotic were reasonabley explained by the shear stress calculation results of left and right common iliac branch vessel walls before and after the lesions. It is shown that the distribution of blood flowing velocity vector, the shear stress at the vessel wall are asymmetrical because of the asymmetrical geometry at the bifurcation of abdominal arterial, so that the shear stress of inner wall is higher than outside. The inner wall shear stress of right common iliac is higher than left. Blood velocity and shear stress increase at the atherosclerotic lesion because the blood vessel becomes narrower, and the blood velocity and shear stress decrease at the downstream of the lesion. It was also shown that the hemodynamic characteristics played a great important role in the occurrence and development of the atherosclerosis diseases at the bifurcation of abdominal arterial. Because of the increase of the shear stress, the atherosclerotic plaque surface will be damaged, leading to tissue hyperplasia. It will be lead to blood cell coacervation that the blood velocity and shear stress decrease at the atherosclerotic plaque downstream.
Aorta, Abdominal
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pathology
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physiopathology
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Atherosclerosis
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pathology
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physiopathology
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Blood Flow Velocity
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physiology
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Finite Element Analysis
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Hemodynamics
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Humans
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Shear Strength
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physiology
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Stress, Mechanical
10.Screening effects of Montreal Cognitive Assessment for sepsis associated encephalopathy
Jiaquan LIU ; Zili ZHANG ; Dequan KONG ; Chengbin YANG ; Yuzhen WU ; Yaben YAO ; Weicheng WU ; Mandong PAN ; Jiyan LIN
Chinese Journal of General Practitioners 2012;11(9):680-682
A total of 192 patients with sepsis were tested by Montreal Cognitive Assessment (MoCA) for a preliminary diagnosis of whether or not there was sepsis associated encephalopathy (SAE) according to their test results.SAE was diagnosed or excluded after consultations and comprehensive analysis on the basis of clinical manifestations and auxiliary examination results.The scores of the patients in this group were (25.7 ± 3.3) points.The sensitivity of MoCA for screening SAE was 0.776 and its specificity 0.963.The rate of diagnostic coincidence between MoCA and comprehensive analysis for SAE was 0.880.The diagnostic concordance between two diagnostic methods of SAE was excellent (kappa value =0.753 ± 0.048,P =0.000).The area under the receiver operating characteristic (ROC) curve of MoCA for screening SAE was 0.929 ± 0.019 (P =0.000) ; the optimal cutoff value was 25.5 points; and its sensitivity was 0.779 and specificity 0.962.And negative correlations existed between score of MoCA and age,disease course and co-existing shock or multiple organ dysfunction syndrome (P < 0.05).