1.Metabolomics analysis of taxadiene producing yeasts.
Huifang YAN ; Mingzhu DING ; Yingjin YUAN
Chinese Journal of Biotechnology 2014;30(2):223-231
In order to study the inherent difference among terpenes producing yeasts from the point of metabolomics, we selected taxadiene producing yeasts as the model system. The changes of cellular metabolites during fermentation log phase of artificial functional yeasts were determined using metabolomics methods. The results represented that compared to W303-1A as a blank control, the metabolites in glycolysis, tricarboxylic acid cycle (TCA) cycle and several amino acids were influenced. And due to the changes of metabolites, the growth of cells was inhibited to a certain extent. Among the metabolites identified, citric acid content in taxadiene producing yeasts changed the most, the decreasing amplitude reached 90% or more. Therefore, citric acid can be a marker metabolite for the future study of artificial functional yeasts. The metabolomics analysis of taxadiene producing yeasts can provide more information in further studies on optimization of terpenes production in heterologous chassis.
Alkenes
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metabolism
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Amino Acids
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metabolism
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Citric Acid
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analysis
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Citric Acid Cycle
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Diterpenes
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metabolism
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Fermentation
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Glycolysis
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Metabolome
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Metabolomics
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Yeasts
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metabolism
2.Research Progress in Heat-clearing Activity of Mineral Drug Gypsum Fibrosum
Liu YANG ; Yisheng ZHANG ; Huifang XU ; Mingyang YUAN ; Zheren LIN
China Pharmacist 2016;19(10):1943-1945
Gypsum Fibrosum is one of mineral medicines, which is widely used in clinics for a long time. It is mainly used for treating epidemic febrile diseases and excess-heat syndrome. The studies on the heat-clearing effects and mechanisms of Gypsum Fibro-sum in recent years were summarized in the paper,which can provide scientific reference for its further research, application and the heat-clearing mechanism exploration.
3.The mechanisms of heparin-derived oligosaccharide on the inhibition of smooth muscle cells proliferation induced by platelet-derived growth factor.
Shuying HE ; Huifang WANG ; Danfeng YU ; Jing YUAN
Acta Pharmaceutica Sinica 2015;50(8):993-9
In this study, the effect of heparin-derived oligosaccharide (HDO) on platelet-derived growth factor (PDGF) induced vascular smooth muscle cells (VSMCs) proliferation and the related signal transduction mechanisms were investigated. MTT assays were used to measure VSMCs proliferation. Cell cycle distribution was analyzed by flow cytometry. The level of key regulatory proteins in PKC, MAPK and Akt/PI3K pathways were determined by RT-PCR, Western blot and immunocytochemical methods. Meanwhile, mRNA expressions of some proto-oncogenes were assayed by RT-PCR method. Our data showed that HDO (0.01, 0.1 and 1 μmol · L(-1)) inhibited 30 ng · mL(-1) PDGF-induced VSMCs proliferation in a dose-dependent manner, blocked the G1/S transition and inhibited the level of key regulatory proteins and some proto-oncogenes (P < 0.05). The results showed that HDO may decrease the key regulatory proteins expression, hence suppress the transcription of proto-oncogene and G1/S transition, finally inhibiting VSMCs proliferation.
4.Ultrasonographic features and histopathological characteristics of primary female genital system lymphoma
Qiang LI ; Shaoling YUAN ; Jianmin ZHAI ; Huifang WANG ; Jing CHEN
Chinese Journal of Ultrasonography 2013;22(7):599-601
Objective To observe the ultrasonographic features of primary female genital system lymphoma and its histopathological characteristics.Methods Sonographic appearances and histopathological characteristics of 14 patients who were pathologically comfirmed as primary female genital system lymphoma were reviewed.Results Fourteen patients underwent ultrasonography and 15 lesions were found,6 cases of which were found in the ovary and cervix separately,3 cases in the uterine body.Among the lesions detected,10 of which respectively represents extremely low echo and homogeneous internal echo,all lymphomas had posterior enhancement.The margins were most frequently circumcribed (13/15),5numbers primary lymphomas of the ovary were mostly elliptical in shape,while 5 numbers of cervix tumors showed the shape of lobular,all of the tumors in the uterine body showed diffuse symmetrical enlargement without disruption of the endometria.Color Doppler imaging showed hypervascularity in most tumors(13/15).The pathological examination showed that all tumors were non-Hodgkin lymphoma diffuse B-cell type.Conclusions Primary female genital system lymphoma has some ultrasonic features,including extremely low echo,homogeneous echo and posterior enhancement.Final diagnosis depends on the histopathology.
5.Correlational study between ultrasonic elastography strain ratio and serum thyroid stimulating hormone on patients with Hashimoto's thyroiditis
Yuxi YAN ; Shaoling YUAN ; Li YANG ; Jianmin ZHAI ; Huifang WANG
Chinese Journal of Ultrasonography 2012;21(5):386-389
ObjectiveTo investigate the value of strain ratio (SR) in the diagnosis of Hashimoto's thyroiditis.Methods 116 patients who had been diagnosed clinically as Hashimoto' s thyroiditis were enrolled in the study.Those patients were classified into four groups:the hyperthyroidism group,the euthyroidism group,the subclinical hypothyroidism group,and the clinical hypothyroidism group.50volunteers with normal thyroid functions were enrolled as a control group.SR of the thyroid and sternocleidomastoid muscle of same side was calculated.The correlation coefficient between the elasiticity SR and the serum thyroid stimulating hormone (TSH) was evaluated and their differences in the diagnosis of Hashimoto's thyroiditis were compared.ResultsThe Spearman's correlation coefficient between the SR and the serum TSH was 0.605,which was significant (P =0.000).The SR increased in an ascending order among the group with hyperthyroidism,the control group,the group with euthyroidism,the group with subclinical hypothyroidism,and the group with clinical hypothyroidism.ConclusionsElastography SR is valuable way for evaluating the progression of Hashimoto's thyroiditis.
6.Regulatory effects of CD44 antibody-A3 D8 on IL-3 Rαand downstream PI3 K/Akt pathway in NB4 cells
Ping CHEN ; Qin YUAN ; Xi JIANG ; Juanying WU ; Huifang HUANG
Chinese Pharmacological Bulletin 2014;(11):1526-1529
Aim To investigate the effect of CD44 anti-body-A3 D8 on the expression of IL-3 Rα and down-stream PI3K/Akt in NB4 cells. Methods The ex-pression of IL-3 Rα mRNA was detected by real-time quantitative RT-PCR, the IL-3Rα protein expression and changes of PI3 K/Akt signal pathway in NB4 cells treated with A3D8 were analyzed by Western blot. An-nexin-V-FITC/PI double staining flow cytometry was u-tilized to detect the apoptotic cells. The inhibitor of PI3 K/Akt signaling LY294002 combined with A3 D8 was used to inhibit the PI3K/Akt in NB4 cells. Re-sults After treated with A3 D8 , both the transcription-al level and translational level of IL-3 Rα were remark-ably reduced, and the PI3K/Akt pathway was inhibi-ted. LY294002 improved the inhibitory and apoptotic effects of A3D8 on NB4 cells. Conclusion CD44 antibody A3 D8 can downregulate the expression of IL-3Rα and inhibit the downstream PI3K/Akt pathway.
7.Effect of the trained SPs by retired nurses in the test of internal medicine inquiry in OSCE for the graduates
Gongxian YUAN ; Huifang CHEN ; Hua WANG ; Yinghui WANG
Chinese Journal of Medical Education Research 2011;10(2):204-206
The article is to discuss the effect of the trained SPs acted by retired nurses in the test of internal medicine inquiry in OSCE for the graduates. The results were evaluated by those samples drawn from medical students who took part in OSCE and the professors. The evaluation of the SPs was objective and accurate, and they acted well. There are remarkable advantages in retired nurses simulating standardized patients, and it deserves extensive application in clinical medical education in our country.
9.Vasorelaxant effect of novel Rho-kinase inhibitors on isolated thoracic aorta rings and underlying mechanisms
Tianyi YUAN ; Subo WANG ; Huifang ZHANG ; Yucai CHEN ; Xiaozhen JIAO ; Ping XIE ; Lianhua FANG ; Guanhua DU
Chinese Pharmacological Bulletin 2016;32(10):1404-1410
Aim To evaluate the vasorelaxant effect of two new chemical entities, J35242 and J35243, on iso-lated rat thoracic aorta rings as Rho-kinase inhibitors, and further to explore the underlying mechanisms of these two compounds. Methods Isolated rat thoracic aorta rings pre-contracted by KCl or norepinephrine ( NE) were used to evaluate the vasodilatory effect of J35242 and J35243 . Through the interventions of sev-eral tool drugs, the mechanisms of compounds concern-ing endothelium, K+ channels and Ca2+ were studied. Results J35242 and J35243 showed potent relaxant effect on both KCl and NE pre-contracted vessels, and exhibited partial endothelium dependency. L-NAME and Methylene Blue( MB) could influence the relaxant effect of these compounds. Meanwhile, the compounds could inhibit intracellular Ca2+ release and extracellu-lar Ca2+ influx, which indicated that the compounds might block the calcium channels to relax the vessels. In addition, the two compounds probably did not dilate the aorta rings through opening potassium channels. Conclusions J35242 and J35243 have vasorelaxant effects on vessels in vitro and the potency of J35242 is stronger than that of J35243 . The underlying mecha-nisms might be endothelium-dependent. Also the com-pounds might block Ca2+ channels, lowering intracel-lular Ca2+ concentration to relax the vessels.
10.Icariin inducting MC3T3-E1 cell proliferation and differentiation with the activation of BMP-2 intracellular signal pathway
Huifang ZHOU ; Nianke SHI ; Yuan XUE ; Zhong YANG ; Chao ZHANG ; Shuo WANG
Chinese Journal of Orthopaedics 2013;(6):664-669
Objective To explore the regulation function of Icariine on the expression of bone morphogenetic protein signaling pathway Smad1,Smad5,Smad4 and to explore the mechanism of promoting MC3T3-E1 cell proliferation and differentiation.Methods MC3T3-E1 cells were treated by 0,10-9,10-8 and 10-7 mol/L Icariine respectively.After stimulated by Icariine 1 d,2 d and 3 d,MTT method and population diploid time were used to observe the cell proliferation,and the cell alkaline phosphatase (ALP) level was assayed.At 21 days later,the alizarin red staining was proceeded.At 1,2 and 3 days later,the RT-PCR was used to detect the mRNA expression level about Smad1,Smad5 and Smad4,and the Western blot was to detect the Smad1,5 and Smad4 protein.At 2 days later,the RT-PCR was used to detect the mRNA expression level about Runx2,BMP-2 and osteoprotegerin (OPG),and the Western blot was used to analyze osteocalcin (OCN) protein level.Results After simulated by Icariine,the proliferation (MTT test),the ALP activity and mineralization of osteoblasts were increased,the cell population diploid was reduced (P<0.05).At 1,2 and 3days later,the results of RT-PCR showed that Icariine continued increasing the mRNA level of Smad1,5 and Smad4 in 10-8 mol/L.At 2 days later,Smad1,Smad5 and Smad4 mRNA expression were obviously reduced in 0 mol/L group,and At 3 days later,Smad1,Smad5 and Smad4 mRNA expression were obviously reduced in 10-9 and 10-7 mol/L groups.At 2 days later,BMP-2,Runx2 and OPG mRNA were obviously increased in 10-8 mol/L group.The results of Western blot showed that OCN,Smad1,Smad5 and Smad4 protein were obviously up-regulated in 10-8 mol/L group.Conclusion Icariine occurs the expressions of BMP-2,Runx2,Smad1,Smad5 and Smad4 by stimulating the bone morphology of MC3T3-E1 cells directly,and promotes the osteogenic differentiation in the manner of expression level synchronous rising.