1.Epidemiological survey of hyperuricemia and gout in Xinjiang adult population
Shan YUAN ; Xiaomei LI ; Yitong MA ; Yining YANG ; Xiang MA ; Ying HUANG ; Zhenyan FU ; Xiang XIE ; Fen LIU ; Lei WANG ; Shuo PAN
Chinese Journal of Endocrinology and Metabolism 2011;27(7):570-572
Four-stage selected random samples were used to analyze the prevalence and distributing feature of hyperuricemia and gout in 13 559 Han,Uighur,and Hazakh adults in Xinjiang. The prevalence of hyperuricemia was 11.00%,3.27%,and 3.94% respectively in Han,Uighur,and Hazakh populations,and 1.32%, 0.65%,and 0.70% for gout,with statistically significant difference among three groups(all P<0.05). No-conditional logistic regression analysis showed that nationality,body mass index,renal function,and serum lipid were risk factors of hyperuricemia,while female and physical activity were protective factors. Eating seafood and animal visceral organs were independent risk factors of hyperuricemia in Han population. Eating animal visceral organs and drinking alcohol were independent risk factors of hyperuricemia in Hazakh population.
2.The expression of interferon-lambda1 in CHO cell.
Wu-Mei YUAN ; Fen-Lian MA ; Qian ZHANG ; Wen-Zhi ZHENG ; Li-Shu ZHENG
Chinese Journal of Experimental and Clinical Virology 2013;27(3):190-192
OBJECTIVETo construct the eukaryotic expression vector PCI-dhfr-lambda1 and PCI-dhfr-SP163-lambda1 which linked the enhancer SP163 with interferon lambda1. Then express the interferon lambda1 in CHO (dhfr-) cells.
METHODSUsing PCR method to introduce the restriction enzyme sites and through the fusion PCR binding the enhancer with the interferon Lambda1. After sequenced, lambda1 and SP163-lambda1 was inserted into PCI-dhfr forming the expression vector PCI-dhfr-lambda1 and PCI-dhfr-SP163-lambda1 which was constructed successfully confirming by sequencing. Then the expressing vectors were transfected into CHO (dhfr-) cells using liposome transfection method and interferon lambda1 protein was assayed with indirect immunofluorescence and Western Blot. Using cytopathic effect inhibition evaluated the antiviral activity of interferon lambda1.
RESULTSSuccessfully constructing the eukaryotic expression vectors of interferon lambda and the vectors could express interferon lambda1. The result of immunofluorescence showed the enhancer developed the expression of interferon lambda1. Detecting the interferon lambda1 in CHO (dhfr-) cells after transfecting 48 hour using Western Blot. The cytopathic effect inhibition showed the expressed interferon lambda1 has the antiviral activity.
CONCLUSIONSuccessfully expressed the interferon lambda1 in CHO (dhfr-) cells and the protein possesses antiviral activity, which may supply a valuable basis for building the stable cell line of interferon lambda1.
Animals ; Blotting, Western ; CHO Cells ; Cricetinae ; Cricetulus ; Fluorescent Antibody Technique, Indirect ; Interleukins ; genetics ; pharmacology ; Polymerase Chain Reaction ; Recombinant Proteins ; biosynthesis ; pharmacology ; Transfection
3.SRAP study on genetic diversity of radix plygoni multiflori in chongqing.
Yuan-hui CHENG ; Chang-hua ZHOU ; Ai-fen MA ; Xiao-gang SHI ; Xing-cui ZHANG
China Journal of Chinese Materia Medica 2007;32(8):661-663
OBJECTIVETo detect the polymorphisms of Radix Plygoni Multiflori in chongqing by means of a new marker system SRAP.
METHODDifferent shaples of Radix Plygoni Multiflori from major production areas were collected. The SRAP was used to asses divergence among 16 populations. The data were analyzed using unweighted pairgroup method, based on arithmetic averages (UPGMA) bootstrap analysis. Cluster analyses was performed by using DPSv3.01 software, the alkaloid was extracted from P. ternate with chlorolform.
RESULT104 combinations generated 250 polymorphie bands, the cluster analysis indicated that 16 materials could be distinguished into two main groups and one special type, Nei&Li similarity coefficient ranged from 0.23-0.99, and the average distance is 0. 44.
CONCLUSIONThe results of the study showed a potential application of SRAP fingerprinting for identification of Radix Plygoni Multiflori.
Cluster Analysis ; DNA, Plant ; genetics ; Genetic Markers ; Genetic Variation ; Nucleic Acid Amplification Techniques ; methods ; Phylogeny ; Plant Roots ; genetics ; Plants, Medicinal ; classification ; genetics ; Polygonum ; classification ; genetics
4.Medium- and long-chain triglyceride propofol reduces the activity of acetyl-coenzyme A carboxylase in hepatic lipid metabolism in HepG2 and Huh7 cells
Li yuan WANG ; Jing WU ; Ya fen GAO ; Duo mao LIN ; Jun MA
The Korean Journal of Physiology and Pharmacology 2020;24(1):19-26
Medium- and long-chain triglyceride (MCT/LCT) propofol is widely used as an intravenous anesthetic, especially in the intensive care unit. The present study aimed to assess whether MCT/LCT propofol is safe in the hyperlipidemic population for long-term use. Free fatty acids (FFAs) were used to establish high-fat stimulation of HepG2 and Huh7 cells. Subsequently, these cells were treated with propofol at the concentration of 0, 4, or 8 µg/ml for 24 and 48 h. The results indicated that the cell viability was notably decreased when the cells were stimulated with 2 mmol/L FFAs and treated with 12 µg/ml MCT/LCT propofol. Accordingly, we chose 2 mmol/L FFAs along with 4 and 8 µg/ml MCT/LCT propofol for the subsequent experiments. Four and 8 µg/ml MCT/LCT propofol inhibited FFA-induced lipid accumulation in the cells and significantly reversed acetyl coenzyme A carboxylase (ACC) activity. In addition, MCT/LCT propofol not only significantly promoted the phosphorylation of AMPK and ACC, but also reversed the FFA-induced decreased phosphorylation of AMPK and ACC. In conclusion, MCT/LCT propofol reverses the negative effects caused by FFAs in HepG2 and Huh7 cells, indicating that MCT/LCT propofol might positively regulate lipid metabolism.
Acetyl-CoA Carboxylase
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AMP-Activated Protein Kinases
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Cell Survival
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Fatty Acids, Nonesterified
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Hepatocytes
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Intensive Care Units
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Lipid Metabolism
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Liver
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Metabolism
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Phosphorylation
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Propofol
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Triglycerides
5.Study on chemical constituents of Inula cappa.
Li-hua ZHENG ; Xiao-jiang HAO ; Chun-mao YUAN ; Lie-jun HUANG ; Jian-xin ZHANG ; Fen DONG ; Tian-yun FAN ; Gui-hui WU ; Yan CHEN ; Yuan MA ; Yi-min FAN ; Wei GU
China Journal of Chinese Materia Medica 2015;40(4):672-678
Column chromatographies over silica gel, Sephadex LH-20, reverse phase C18, and MCI, and semi-preparative HPLC were used for separation and purification of constituents from Inula cappa. The 22 compounds were obtained and their strutures were determined by NMR and MS spectra data as nine flavonoids: luteolin (1), apigenin (2), chrysoeriol (3), artemetin (4), 2', 5-di- hydroxy-3, 6, 7, 4', 5'-pentamethoxyflavone (5), chrysosplenol C (6), apigenin-5-0-β-D-glucopyranoside (7), luteolin-3-methyl, luteolin-3-methylether-4'-0-β-D-glucopyranoside (8), luteolin-4'-0-β-D-glucopyranoside (9); four triterpenes: darma-20, 24-dien- 3β-0-acetate (10), darma-20, 24-dien-3β-ol (11), epirfiedelanol (12), friedelin (13); three coumarins: scopoletin (14) , isosco- poletin (15) , scopolin(16) , and other types of compounds stigmasta-5, 22-dien-3β-0-7-one (17), stigmasterol (18), palmitic acid (19), linoleic acid (20), linoleic acid methyl ester (21), (E) -9, 12, 13-trihydroxyoetadee-10-enoie acid (22). Compound 5 is a new natural product. Compounds 3-9, 15, 17, 21, and 22 were isolated from this genus for the first time.
Drugs, Chinese Herbal
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chemistry
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isolation & purification
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Inula
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chemistry
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Molecular Structure
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Spectrometry, Mass, Electrospray Ionization
6.Association between apolipoprotein A5 gene polymorphism and coronary heart disease in the Han population from Xinjiang
Shan YUAN ; Yi-Tong MA ; Xiang XIE ; Yi-Ning YANG ; Zhen-Yan FU ; Xiang MA ; Xiao-Mei LI ; Fen LIU ; Bang-Dang CHEN
Chinese Journal of Epidemiology 2011;32(1):51-54
Objective The aim is to investigate the association between coronary heart disease (CHD) and c.553G>T polymorphism of apolipoprotein A5 (ApoA5) gene and the influence of serum lipid level in the Hah ethnic population of Xinjiang. Methods The polymorphism of ApoA5 gene in 486 patients with CHD and 501 controls was analyzed by methods of polymerase chain reaction and restriction fragment length polymorphism analysis. Level of serum lipid in each patient was detected at the same time. Results There was significant difference in the distribution of genotypes between CHD group and controls group ( x2 = 8.757, P= 0.013 ). Non-conditioned logistic regression analyses, after adjusted for age, gender, smoking, total serum cholesterol, presence of hypertension and diabetes, revealed that individuals who carried T allele (TT + GT genotype) had an increased risk of CHD, compared to GG genotype (OR= 1.753,95%CI: 1.030-2.983, P<0.05 ). There was also a remarkable difference noticed in the level of serum triglyceride by genotypes in CHD group and control group (t=5.242, P<0.01; t=-3.499, P=0.001 ). Individuals in the two groups who carried T allele had higher level of serum triglyceride than those carried GG genotype. Individuals in CHD group who carried T allele had higher level of serum total cholesterol than those carried GG genotype (t=-2.465, P=0.014). Conclusion It seemed that the c.553G>T polymorphism of ApoA5 gene had influenced on the level of serum triglyceride and the total cholesterol among Han population in Xinjiang. c.553G>T polymorphism was associated with the development of CHD, while T allele might be an influencing risk factor on CHD.
7.Immunoregulation effects in vitro of the xenoprotein in combination with recombinant human granulocyte-macrophage colony stimulating factor and bacillus Calmette-Guerin.
Ming-Li WANG ; Zhi-Gang XIE ; Han LU ; Ming SHI ; Mei-Ru HU ; Ming YU ; Yuan-Fang MA ; Bei-Fen SHEN ; Ning GUO
Journal of Experimental Hematology 2008;16(6):1408-1412
This study was aimed to investigate the effects of xenogeneic antigen neu-Fc in combination with the recombinant human granulocyte-macrophage colony stimulating factor (GM-CSF) and Bacillus Calmette-Guerin (BCG) on the regulation of Th1 and Th2 immune response in vitro. The rat neu L2-S2 domain was engineered as a chimeric protein with human IgG Fc. The eukaryotic expression vector was constructed. The recombinant protein was stably expressed in CHO cells and purified by rProtein A Sepharose Fast Flow column. The recombinant protein was identified by SDS-PAGE and Western blot. Peripheral blood mononuclear cells (PBMNCs) were obtained by means of standard Ficoll separation from the blood of healthy donors. Neu-Fc-induced PBMNC proliferation was tested by MTT. The production of IL-12 and IL-10 was measured by ELISA. The results showed that the level of IL-12 decreased and IL-10 increased after PBMNCs were incubated with MCF-7 cultural supernatant. 10 nmol/L neu-Fc strongly induced the cell proliferation. Compared with neu-Fc or GM-CSF or BCG treatment alone, neu-Fc in combination with GM-CSF and BCG significantly stimulated IL-12 production and inhibited IL-10 production (p < 0.01). It is concluded that the neu-Fc can stimulate the proliferation activity of PBMNCs. neu-Fc, GM-CSF and BCG costimulation efficiently induces Th1 immune response.
Animals
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BCG Vaccine
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immunology
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CHO Cells
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Cricetinae
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Cricetulus
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Granulocyte-Macrophage Colony-Stimulating Factor
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immunology
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Humans
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Interleukin-10
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metabolism
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Interleukin-12
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metabolism
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Rats
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Recombinant Proteins
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immunology
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Th1 Cells
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immunology
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Th2 Cells
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immunology
8.Preparation of the monoclonal antibody against human Bocavirus VP2 protein.
Zhi-Hui ZHAO ; Peng-Hao XUE ; Jian-Min WEI ; Qian ZHANG ; Wen-Zhi ZHENG ; Fen-Lian MA ; Wu-Mei YUAN ; Li-Shu ZHENG
Chinese Journal of Experimental and Clinical Virology 2012;26(1):63-65
OBJECTIVETo express and purify HBoV VP2 protein, and the monoclonal antibody against HBoV VP2 protein was prepared with hybridoma technique.
METHODSThe HBoV VP2 cloned into vector pET-30a was expressed in E. coil. After purified by immobilized metal affinity chromatography, the BALB/c mouse was immunized with purified protein as antigen. The positive hybridoma cells were screened with hybridoma technique and ELISA assay. Isotype and titer of the monoclonal antibody were detected.
RESULTSThe recombinant HBoV VP2 protein was expressed and purified, and then the monoclonal antibody was obtained with hybridoma technique. The titer of the IgG monoclonal antibody was up to 1:4 x 10(5).
CONCLUSIONMonoclonal antibody against recombinant HBoV VP2 protein was prepared and the antibody titer was high. This work may provide a new method in rapid diagnosis and study of HBoV.
Animals ; Antibodies, Monoclonal ; immunology ; Capsid Proteins ; genetics ; immunology ; Human bocavirus ; immunology ; Hybridomas ; Mice ; Mice, Inbred BALB C ; Plasmids ; Recombinant Proteins ; biosynthesis ; immunology ; isolation & purification
9.Characteristics of microsomal phase II metabolic enzymes in mouse embryonic stem cell-derived liver tissue.
Tong LI ; Mei-Yuan GUO ; Kui-Fen MA ; Yue DU ; Liang-Yan HE ; Dan-Yan ZHU ; Yi-Jia LOU
Journal of Zhejiang University. Medical sciences 2013;42(5):530-537
OBJECTIVETo investigate the characteristics of phase II metabolic enzymes in mouse embryonic stem (ES) cell-derived liver tissue.
METHODSMature hepatocytes were differentiated from embryonic stem cells in cultured mouse embryoid bodies (EB) at d18. Western blot was used to detect the expression of uridine 5'-diphosphate glucronosyl transferase (UGT1a1,UGT1a6) and microsomal glutathione S-transferases 1(mGST1) during the differentiation course.The derived liver tissue was incubated with UDPGA and 7-HFC,the formation of 7-HFC glucuronide was detected by HPLC to examine the total activities of UGT1a1 and UGT1a6. Furthermore, the microsomes were incubated with CDNB and GSH,and the mGST1 activity was measured by spectrometry.
RESULTSAn increase tendency of UGT1a1 expression was noticed during the differentiation course. UGT1a6 and mGST1 were not detected in the earlier stage until d18 of differentiation. The metabolic activity of mGST1 in the derived hepatocytes was 7.65 nmol/min/mg on d18.
CONCLUSIONThe ES cell-derived liver tissue possesses partial metabolic function of phase II enzymes on d18 of differentiation,which might be used as a model for in vitro research on hepatic pathophysiology and phase II drug metabolism.
Animals ; Cell Differentiation ; Embryoid Bodies ; cytology ; Embryonic Stem Cells ; cytology ; Glucuronosyltransferase ; physiology ; Glutathione Transferase ; physiology ; Hepatocytes ; cytology ; enzymology ; Mice
10.A study on the effect of the chitosan thermosensitive hydrogel loading recombinant human bone morphogenetic protein-2 on repairing periodontal defects.
Zhi-wei MA ; Yong-jie ZHANG ; Zhi-fen WU ; Rong WANG ; Hao ZHU ; Yuan LI ; Jie XU ; Qing LIU
West China Journal of Stomatology 2008;26(1):23-26
OBJECTIVETo observe the effect of the chitosan thermosensitive hydrogel loading recombinant human bone morphogenetic protein-2 (rhBMP-2) on repairing periodontal defects.
METHODSTo prepare artificial furcation defects model in the posterior area in 3 healthy male dogs, and then to inject chitosan thermosensitive hydrogel loading of rhBMP-2 after fast suturing tissue flap. The groups filled with nothing or filled only with chitosan thermosensitive hydrogel were the controls. The dogs were sacrificed after 5 weeks and the periodontal regeneration was observed histologically.
RESULTSThe histological observation showed that the chitosan thermosensitive hydrogel loading rhBMP-2 group achieved apparent periodontal tissue regeneration occupying the majority of the defects and the control groups got only a small amount of periodontal tissue regeneration.
CONCLUSIONThe chitosan thermosensitive hydrogel loading rhBMP-2 can effectively promote the periodontal tissue regeneration, while simplifying the surgical operation. It might be a potential means for periodontal regeneration.
Animals ; Bone Morphogenetic Protein 2 ; Bone Morphogenetic Proteins ; Chitosan ; Dogs ; Humans ; Hydrogel, Polyethylene Glycol Dimethacrylate ; Male ; Recombinant Proteins ; Regeneration ; Transforming Growth Factor beta