1.Preparation of titanium substrate biomaterials by using microarc oxidation and measurement of blood compatibility
Cheng YANG ; Lie MENG ; Ting CHANG ; Yuan TIAN
Chinese Journal of Tissue Engineering Research 2007;11(31):6315-6317
BACKGROUND: Microarc oxidation (MAO) is a break-through anodyzing technology for forming oxide films on valve metal.Use of this technology allows thick, porous oxide layers to be formed on the surface of pure titanium. Few biocompatibility reports using this treatment have been found.OBJECTIVE: The blood compatibility of a novel surface modified titanium substrata biomaterial using MAO was investigated.DESIGN: Positive and negative control, contrast observation and gold standard control.SETTING: Wuhan Union Hospital.MATERIALS: A healthy male adult New-Zealand rabbit, weighing 2.5 kg and ordinary grade, was selected in this study.Pure titanium sticks TA1 (Baoji Yingnaite Non-ferrous Metal Co., Ltd.), MAO-Ti and 20 g/L potassium oxalate were also selected in this study.METHODS: The study was carried out in the Laboratory of General Surgery, Union Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology in May 2006. ① Materials: Titanium substrate of 10 mm in diameter and 2 mm in depth was put in an electrolyte which was quipped with deionized water, dibasic sodium phosphate, and ethanoic acid calcium for MAO treatment for 10 minutes. ② Groups: Three groups were analysed: test group, negative control group and positive control group. Test group: MAO-Ti was dipped in 10 mL saline; Positive control group: 10 mL deionized water was added in each tube; Negative control group: 10 mL saline was added in each tube. ③ Operation: Fresh whole blood was collected from rabbit and then mixed with the liquids in the three groups respectively after anti-coagulation. In addition, UV-Visible Spectrophotometer was used to evaluate the hemolytic ratio. A hemolytic ratio below or equal to 5% indicated that this novel material fitted the requirements. On the contrary, a hemolytic ratio higher than 5% proofed the existence of a hemolyzation.MAIN OUTCOME MEASURES: The hemolytic ratio of materials in three groups.RESULTS: The hemolytic ratio of the test group was 0.90%. The result indicated that this new material had no haemolysis effect.CONCLUSION: The material does not resolve red blood cells and is coincident with the international and governmental standard.
2.Analysis on morphological characteristics, photosynthetic characteristics and chemical constituents of Inula lineariifolia from different populations.
Feng-chen CHEN ; Chang-lin WANG ; Qiao-sheng GUO ; Xin TIAN ; Yuan-yuan QIU
China Journal of Chinese Materia Medica 2015;40(22):4389-4394
Twelve populations of Inula lineariifolia were used as materials to measure morphological characteristics, photosynthetic parameters and chemical constituents. It aims to provide a theoretical basis for germplasm resources evaluation. The results showed that I. lineariifolia had relatively rich morphological diversity, there were significant differences of morphological characteristics, photosynthetic parameters and chemical constituents among populations. There was positive correlation on morphological characteristics and P(n). Twelve populations were divided into three-type. The three populations of Xuyi, Mingguang and Fengyang were of narrower-longer leaf, bigger biomass,better photosynthetic and higher chemical constituents. Then they were classified for a similar group. It proved that the three populations were more suitable for cultivation and promotion.
Biomass
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China
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Flowers
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chemistry
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growth & development
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metabolism
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Inula
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chemistry
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classification
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growth & development
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metabolism
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Photosynthesis
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Plant Leaves
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chemistry
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growth & development
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metabolism
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Plant Stems
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chemistry
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growth & development
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metabolism
3.Inhibitory effects of ethyl acetate extract of Huanglian Jiedu decoction on hyphae development of Candida albicans.
Tian-ming WANG ; Yuan-yuan YAN ; Gao-xiang SHI ; Dan XIA ; Jing SHAO ; Chang-zhong WANG
China Journal of Chinese Materia Medica 2014;39(24):4834-4838
OBJECTIVETo investigate the effects of ethyl acetate extract of Huanglian Jiedu decoction (EAHD) on hyphae development of Candida albicans.
METHODInverted microscope, fluorescence microscope, SEM were applied to inspect the Morphological change of C. albicans treated by EAHD at different concentrations. Solid agar plate was utilized to evaluate the colony morphology. Quantitative Real-ime PCR(qRT-PCR) was adopted to observe the expression of hyphae-specific genes such as HWP1, ALS3, UME6, CSH1, SUN41, CaPDE2.
RESULTEAHD with concentration of 312 and 1 250 mg . L-1 could inhibit formation of hyphae and colony morphology. The expression of HWP1, ALS3, UME6, CSH1 were downregulated 4. 13, 3. 64, 2. 46, 2. 75 folds ,while the expression of SUN41 were upregulated 7. 26 folds, CaPDE2 keep unchanged.
CONCLUSIONEAHD could inhibit formation of hyphae and colony morphologies of C. albicans through downregulating HWP1, ALS3, UME6 and CSH1.
Acetates ; Biofilms ; drug effects ; growth & development ; Candida albicans ; cytology ; drug effects ; genetics ; growth & development ; Down-Regulation ; Drugs, Chinese Herbal ; pharmacology ; Fungal Proteins ; genetics ; Gene Expression Regulation, Fungal ; drug effects ; Hyphae ; Medicine, Chinese Traditional ; Microscopy, Fluorescence ; Reverse Transcriptase Polymerase Chain Reaction
4.Effect and mechanism of recombinant hirudin on atherosclerotic plaques in apolipoprotein E knockout (ApoE(-/-)) mice.
Jin-fan TIAN ; Chang-Jiang GE ; Shu-zheng LU ; Fei YUAN ; Kang ZHAO
Chinese Journal of Integrated Traditional and Western Medicine 2015;35(2):198-203
OBJECTIVETo explore the effect and mechanism of hirudin on atherosclerotic plaques in apolipoprotein E knockout (ApoE(-/-)) mice.
METHODSTotally 24 ApoE(-/-) mice, 7-8 weeks old were fed with high fat diets. They were randomly divided into the recombinant hirudin treatment group (drug group) and the model group according to body weight and different dens, 12 in each group. Twelve C57BL/6J mice, 7-8 weeks old fed with high fat diet were recruited as the normal control group. Recombinant hirudin (0.25 mg/kg) was intraperitoneally injected to mice in the drug group from the 10th week old once every other day for five successive weeks. Equal volume of normal saline was injected to mice in the model group. Mice in the normal control group received no treatment. All mice were sacrificed after fed with high fat diet until they were 20 weeks old. Serum levels of total cholesterol (TC), triglyceride (TG), low-density lipoprotein (LDL), high-density lipoprotein (HDL), high-sensitive C-reactive protein (hs-CRP), E-selectin, interleukin-6 (IL-6), and stromal metalloproteinase-2 (MMP-2) were detected. The plaque/lumen area and extracellular lipid composition/ plaque area were analyzed by HE staining and morphometry. Changes of signaling molecules in store-operated calcium channels, including stromal interacting molecule 1 (STIM1), Orail protein, and transient receptor potential channel 1 (TRPC1) were determined by Western blot. Results Lipid plaque formed in the aorta vessel wall of 20-week old mice in the model group. Compared with the normal control group, serum levels of TC, TG and LDL increased (P<0.01), hs-CRP, E-selction, IL-6, and MMP-2 obviously increased (P<0.01, P<0.05) in the model group; expression levels of STIM1, TRPC1, and Orail significantly increased (P<0.01). Compared with the model group, the plaque/lumen area and the extracellular lipid composition/plaque area significantly decreased in the drug group (P<0.05, P<0.01); serum levels of TC and LDL, hs-CRP, E-selction, IL-6, and MMP-2 obviously decreased (P<0.05, P<0.01); expression levels of STIM1, TRPC1, and Orail were significantly down-regulated (P<0.05, P<0.01).
CONCLUSIONHirudin could significantly improve lipids and endothelial functions of ApoE(-/-) mice, down-regulate expression levels of STIM1, Orai1, and TRPC1, and thus delaying the occurrence and development of atherosclerosis.
Animals ; Aorta ; Apolipoproteins E ; metabolism ; Atherosclerosis ; C-Reactive Protein ; Cholesterol ; Diet, High-Fat ; Drugs, Chinese Herbal ; E-Selectin ; Hirudins ; metabolism ; Interleukin-6 ; Lipids ; Lipoproteins, HDL ; Lipoproteins, LDL ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Plaque, Atherosclerotic ; metabolism ; Recombinant Proteins ; metabolism ; Triglycerides
5.Effect of andrographolide on Candida albicans biofilm dispersion.
Gao-Xiang SHI ; Yuan-Yuan YAN ; Jing SHAO ; Ke-Qiao LU ; Meng-Xiang ZHANG ; Tian-Ming WANG ; Chang-Zhong WANG
China Journal of Chinese Materia Medica 2014;39(17):3339-3343
Along with the increase in fungal infections, Candida albicans prevention and control become the focus of anti-fungal infection at present. This study aims to discuss the effect monomer andrographolide (AG) on C. albicans biofilm dispersion. In the experiment, micro-well plates and medical catheter pieces were used to establish the C. albicans biofilm model. It was discovered by XTT assay and flat band method that 1 000, 500, 250 mg x L(-1) AG could impact the activity of C. albicans biofilm dispersion cells. The morphological structures of residual biofilms on catheter pieces were observed with scanning electron microscopy, which showed that 1 000, 500, 250 mg x L(-1) AG could induce C. albicans biofilm dispersion in a dose-dependent manner, and the dispersed cells were dominated by the yeast phase. According to the real-time fluorescence quantification PCR (qRT-PCR) test, AG could up-regulate HSP90 expression and down-regulate UME6 and PES1 expressions. This study demonstrates that AG could induce C. albicans biofilm dispersion to some extent.
Anti-Inflammatory Agents
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pharmacology
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Biofilms
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drug effects
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Candida albicans
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genetics
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physiology
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ultrastructure
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Diterpenes
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pharmacology
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Dose-Response Relationship, Drug
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Fungal Proteins
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genetics
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Gene Expression Regulation, Fungal
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drug effects
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HSP90 Heat-Shock Proteins
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genetics
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Microscopy, Electron, Scanning
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Reverse Transcriptase Polymerase Chain Reaction
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Time Factors
6.Effect of andrographolide derivative Yanhuning on in vivo Candida albicans biofilms in rats.
Gao-Xiang SHI ; Yuan-Yuan YAN ; Jing SHAO ; Meng-Xiang ZHANG ; Ke-Qiao LU ; Tian-Ming WANG ; Chang-Zhong WANG
China Journal of Chinese Materia Medica 2014;39(15):2924-2929
OBJECTIVETo investigate the effect of andrographolide derivative Yanhuning (YHN) on Candida albicans biofilms in rats.
METHODThe rat C. albicans biofilms subcutaneous catheter model was established by intraperitoneally injecting YHN (40, 20, 10, 5, 2.5 mg x kg (-1)), with the FLC (80 mg x kg(-1)) positive group as the control group. After 7 d, CFU counting and XTT assay were used to evaluate the effect of YHN on C. albicans biofllms in vivo. Scanning electron microscopy (SEM) was applied to observe the morphological changes in rat biofilms intervened by YHN. The real-time fluorescence quantification PCR was adopted to detect expressions of C. albicans adhesion-related genes, such as ALS1, ALS3, HWP1, EAP1 and MP65.
RESULTThe YHN group showed much less CFUs on catheter pieces and lower XTT metabolic activity than the blank group, with dosage dependence. SEM also showed that YHN could obviously decrease C. albicans adhesion on subcutaneous catheters in rats. According to qRT-PCR's results, YHN can down-regulate expressions of ALS1, ALS3, HWP1, EAP1 and MP65.
CONCLUSIONYHN could inhibit C. albicans biofilms in rats.
Animals ; Biofilms ; drug effects ; growth & development ; Candida albicans ; cytology ; drug effects ; physiology ; Catheters ; microbiology ; Cell Adhesion ; drug effects ; Diterpenes ; chemistry ; pharmacology ; Dose-Response Relationship, Drug ; Rats
7.Effect of Huanglian Jiedu decoction in combination with fluconazole on ergosterol of fluconazole-resistant Candida albicans.
Yuan-yuan YAN ; Tian-ming WANG ; Gao-xiang SHI ; Meng-xiang ZHANG ; Ke-qiao LU ; Jing SHAO ; Chang-zhong WANG
China Journal of Chinese Materia Medica 2015;40(4):727-732
OBJECTIVETo investigate the effects of ethyl acetate extract of Huanglian Jiedu decoction (EAHD) , alone and in combination with fluconazole (FLZ) on FLZ-resistant Candida albicans.
METHODThe minimum inhibitory concentrations (MIC) and sessile MIC80 (SMIC80) of EAHD and FLZ to FLZ-resistant C. albicans were determined by CLSI M27-A3 microdilution method, and the synergy of EAHD combined with FLZ were examined by the checkerboard microdilution assay. Agar plate-method was adopted to observe the rate of antifungal activity according to time-kill curve. HPLC and qRT-PCR were utilized to evaluate the changes of ergosterol content and expressions of related genes, respectively.
RESULTMICs of EAHD ranged from 156 to 1,250 mg · L(-1), those of FLZ from 256 to above 2,048 mg · L(-1) with FICI approximate 0.066 in combination; SMIC80 of EAHD were higher than 1,250 mg · L(-1), SMIC80 of FLZ were higher than 512 mg · L(-1) and up to above 2,048 mg · L(-1). Combination group also showed synergy effect except one group showing addition effect. The results of T-K experiment also confirmed obviously fungicidal effect when treated for 12 h. When compared with control groups, the ergosterol was reduced 85% and 50% in the treatments of combination and EAHD alone by HPLC, respective- ly. The expressions of ERG1, ERG2, ERG6, ERG7 and ERG11 were upregulated, and ACS1, ACS2, MET6 were downregulated when exposed to FLZ. The expressions of the above genes were downregulated by treatment of EAHD. The expressions of ERG2, ERG6, ERG11 were upregulated, while ERG1, ERG7, ACS1, ACS2, MET6 were downregulated in combination group.
CONCLUSIONThe combination of EAHD and FLZ exhibited synergy against FLZ-resistant C. albicans through decreasing the synthesis of ergosterol, and resulting in the breakage of cell membrane.
Antifungal Agents ; pharmacology ; Candida albicans ; drug effects ; growth & development ; metabolism ; Drug Resistance, Fungal ; drug effects ; Drug Synergism ; Drugs, Chinese Herbal ; pharmacology ; Ergosterol ; biosynthesis ; Fluconazole ; pharmacology ; Microbial Sensitivity Tests
8.Anti-attachment effect of ethyl acetate extract of Huanglian Jiedu decoction on Candida glabrata.
Tian-ming WANG ; Meng-xiang ZHANG ; Gao-xiang SHI ; Yuan-yuan YAN ; Jing SHAO ; Dan XIA ; Chang-zhong WANG
China Journal of Chinese Materia Medica 2015;40(3):516-521
OBJECTIVETo investigate anti-attachment effect of ethyl acetate extract of Huanglian Jiedu decoction (EAHD) on Candida glabrata.
METHODSerial 2-fold dilution assay was used to determine the minimum inhibitory concentrations MICs of EAHD to C. glabrata. XTT assay was used to evaluate the effect of EAHD against adhesion of C. glabrata. Inverted microscope, scanning electron microscope (SEM) and fluorescein diacetate (FDA) staining were applied to observe the morphological changes of C. glabrata in adhesion. PCR was adopted to inspect the expression of attachment-related genes such as EPA1, EPA6 and EPA7.
RESULTThe MIC of EAHD and fluconazole to C. glabrata were 320 mg · L(-1) and 1 mg · L(-1) respectively. The total cells including budding cells decreased in a dose-dependent manner following EAHD treatment. The expressions of EPA1, EPA6 and EPA7 were downregulated dramatically after EAHD treatment.
CONCLUSIONEAHD could effectively inhibit adherence of C. glabrata.
Acetates ; Candida glabrata ; drug effects ; physiology ; Drugs, Chinese Herbal ; pharmacology ; Fungal Proteins ; genetics ; Lectins ; genetics ; Plant Extracts ; pharmacology
9.Clinical analysis of rituximab combined with chemotherapy for treatment of diffuse large B-cell lymphoma
Hui LIU ; Naibai CHANG ; Jianping WEI ; Shengming ZHAO ; Yun FAN ; Yeping ZHANG ; Jiangtao LI ; Ru FENG ; Wei CHENG ; Yuan TIAN
Journal of Leukemia & Lymphoma 2009;18(3):152-154
Objectives To evaluate the efficacy of rituximab combined with chemotherapy in the treatment of diffuse large B-cell lymphoma (DLBCL) and the relationship of clinical prognosis with the International Prognostic Index (IPI) by the using rituximab in autologous peripheral stem cell transplantation (APBSCT) for the patients of DLBCL. Methods 21 patients with DLBCL, 11 patients of them were at IPI low risk, and 3 patients were IPI at low intermediate risk, 3 patients were at IPI high intermediate risk, 4patients IPI high risk. Rituximab combined with CHOP regimen (cyclophosphamide, adriamycin, vincfistine and prednisone) was given for 4~8 courses. 5 patients received APBSCT. The mobilizing regimen was rituximab combined with cyclophosphamide(CTX) and etoposide(VP16). The conditioning regimen were CBV(CTX combined with VP16 and carmustine). Results In 21 patients, the complete response rate was 61.9 %,with overall response rate 90.5 %. 2-year progression free survival was (69.74±10.43)%. 2-year overall survival was (84.44:1:8.35) %. The complete response rate was 92.9 % and overall response rate was 100 % in the patients IPI≤2. The overall response rate was 71.4 % in the patients with IPI≥3. The complete response rate was higher in the patients with IPI≤ 2 (P<0.01). The amount of mononuclear cells (M NC) in harvest were 7.34 (4.6~8.53)×108/kg. The CD+34 cells in harvest were 8.82 (2.1~10.34)×1O6/kg. The mean time of neutrephil recovering to 0.5×109/L after APBSCT was +9 day. The mean time of platelet recovering to 20×109/L after APBSCT was +12 day. The major adverse reaction were infusion related response (14.3 %) and hematological toxieities. Conclusion The efficacy of rituximab combined with chemotherapy in the treatment of DLBCL is effective, The complete response rate was higher in the patients with IPI≤2 than in the patients with IPI≥3.Using rituximab in mobilizing regimen, all patients had harvested enough CD+34 cells. Rituximab given at +1day did not affect the hematopoiesis reconstruction.