1.Study on Tissue Culture Techniques for Radicle of Bletilla Striata
Xiang LEI ; Mengyao HUANG ; Yanxia CHANG ; Yuanyuan JING ; Keli CHEN
China Pharmacist 2014;(4):613-614
Objective:To seek the best part and optimal culture medium for the tissue culture of Bletilla striata. Methods: The effects of hormone concentration and proportion on the induced differentiation, propagation and rootage of Bletilla striata were investiga-ted. Results:The radicle of Bletilla striata was the best part to induce the clustered shoots with the optimal culture medium of 1/2MS+1. 0 mg·L-1 6-BA+2. 0 mg·L-1 NAA. The best hormone concentration for inducing the clustered shoots was MS+1. 0 mg·-1 L 6-BA+0. 05 mg·L-1 NAA, and the optimal rooting medium was 1/2MS+0. 5 mg·L-1 NAA. Conclusion: The tissue culture system for Bletilla striata is established.
2.Effects of transforming growth factor-β on the expressions of epithelial-to-mesenchymal transition-related molecules in A431 cells
Yuanyuan GUO ; Bin PENG ; Guiqiong XIANG ; Songmei GENG
Chinese Journal of Dermatology 2014;47(7):490-493
Objective To evaluate the effect of transforming growth factor-β (TGF-β) on the expressions of epithelial-to-mesenchymal transition (EMT)-related molecules in human A431 epidermal squamous cell carcinoma cells.Methods Cultured A431 cells were classified into two groups:TGF-β group treated with TGF-β of 7.5 μg/L for 72 hours,and control group remaining untreated.Subsequently,cell morphology was observed under an inverted microscope,and real time-PCR and Western blot were performed to quantify the mRNA and protein expressions of EMT-related molecules including E-cadherin,N-cadherin,vimentin and β-catenin respectively.Results After TGF-β treatment,the cells became dispersed and spindle in shape.The mRNA expression levels of E-cadherin,Ncadherin,vimentin,and β-catenin in TGF-β group were 0.317,2.475,11.340 and 2.615 folds those in the control group respectively.Western blot showed a marked increase in the expression of N-cadherin,vimentin,and β-catenin proteins but a notable decrease in that of E-cadherin in the TGF-β group compared with the control group.Conclusions TGF-β can induce EMT in A431 cells,and increased expression of TGF-β may contribute to the invasion and metastasis of SCC.
3.The effect of Brandt-Daroff exercises on positional vertigo with atypical positioning nystagmus
Xin MA ; Xiang ZHANG ; Yuanyuan JING ; Ruiming XIA ; Lisheng YU
Chinese Archives of Otolaryngology-Head and Neck Surgery 2006;0(08):-
OBJECTIVE Identify the effect of Brandt-Daroff exercises for positional vertigo with atypical positioning nystagmus.METHODS 11 cases of positional vertigo during 2006~2007 with atypical positioning nystagmus when carrying Dix-Hallpike and roll test,the nystagmus had the follwing characters:poly-positions,atypical direction,long lasting(more than 3 mins),no fatigue.After ruling out centrul neural system disorders,the patients were instructed to do Brandt-Daroff exercises 2-3 times/day,and 6-10 times every time.RESULTS After doing the exercises for 3-4 days,the vertigo of all the 11 cases were all relieved.After 1-2 weeks,10 cases were cured,syptom of the other 1 was relieved significantly,and the nystagmus was alleviated.CONCLUSION Cases with atypical positioning nystagmus may present a different kind of BPPV related with the otolith and vestibular nerve disease.Brandt-Daroff exercises was a good method for the atypical positioning nystagmus cases.
4.Determination of Depolymerization Products in Supercritic al Methanolysis of Poly(ethylene terephthalate)polyester by Reversed Phase High Performance Liquid Chromatography
Yong YANG ; Changming ZHANG ; Yuanyuan XU ; Yongwang LI ; Hongwei XIANG
Chinese Journal of Analytical Chemistry 2001;29(6):676-678
qualitative and quantitative analysis of solid depolymerization products for supercritical methanolysis of ploy(ethylene terephthalate) polyester (PET) were studied by means of reversedphase high performance liquid chromatogr aphy on ZorbaxC8(4.6mm×250mm) with 70%(V/V) methanol as mobile phase. The results showed that the solid depolymerization products were mainly composed of dimethyl terephthalate (DMT), methyl(2hydroxyethyl)terephthalate (MHET), bis(hydroxyethyl)tere phthalate (BHET), dimers and oligomers, which could be separated effectively under the above chromatographic conditions. The calibration curves of DMT and BHET were linear in the range of 1.0~45 mg/L (n=8),r=0.9998~1.0000 and RSD<2.8%. The determination limits of DMT and BHET were 4.0×10-4μg and 6.0×10-4μg respectively.
5.Expression of NADPH oxidase and production of reactive oxygen species in aorta in an active immunization mouse model with AT1-EC2 peptide.
Yumiao, WEI ; Yaoqi, CHEN ; Zhi, LI ; Wenping, ZHOU ; Yuanyuan, LV ; Zihua, ZHOU ; Xiang, CHENG ; Yuhua, LIAO
Journal of Huazhong University of Science and Technology (Medical Sciences) 2012;32(4):490-4
The antibody against AT1-EC2 plays a role in some kinds of inflammatory vascular diseases including malignant hypertension, preeclampsia, and renal-allograft rejection, but the detailed mechanisms remain unclear. In order to investigate the changes of NADPH oxidase and reactive oxygen species in the aorta in a mouse model which can produce AT1-EC2 antibody by active immunization with AT1-EC2 peptide, 15 mice were divided into three groups: control group, AT1-EC2-immunized group, and AT1-EC2-immunized and valsartan-treated group. In AT1-EC2-immunized group and AT1-EC2-immunized and valsartan-treated group, the mice were immunized by 50 μg peptide subcutaneously at multiple points for 4 times: 0, 5, 10, and 15 days after the experiment. In AT1-EC2-immunized and valsartan-treated group, valsartan was given at a dose of 100 mg/kg every day for 20 days. After the experiment, the mice were sacrificed under anesthesia and the aortas were obtained and frozen in liquid nitrogen for the preparation of frozen section slides and other experiments. The titer of AT1-EC2 was assayed by using ELISA. The level of NOX1 mRNA in the aorta was determined by using RT-PCR. The expression of NOX1 was detected by using Western blotting. Confocal scanning microscopy was used to assay the α-actin and NOX1 expression in the aortic tissue. The O(2)∸ production was detected in situ after DHE staining. The mice produced high level antibody against AT1-EC2 in AT1-EC2-immunized group and AT1-EC2-immunized and valsartan-treated group, and the level of NOX1 mRNA in the aortic tissues was 1.6±0.4 times higher and the NOX1 protein expression was higher in AT1-EC2-immunized group than in control group. There were no significant differences in the level of NOX1 mRNA and protein expression between control group and AT1-EC2-immunized and valsartan-treated group. The expression and co-localization of α-actin and NOX1 in AT1-EC2-immunized group increased significantly as compared with those in control group, and the O(2)∸ production increased about 2.7 times as compared with control group. There were no significant differences between control group and AT1-EC2-immunized and valsartan-treated group. It is concluded that active immunization with AT1-EC2 can activate NOX1-ROS, and increase vascular inflammation, which can be inhibited by AT1 receptor blocker valsartan. This may partially explain the mechanism of the pathogenesis of inflammatory vascular diseases related to antibody against AT1-EC2.
6.Establishment of cell-based screening system for compound regulating the stability of retinoic acid receptors
Bo JING ; Penghui CHEN ; Xiang GAO ; Yuanyuan XU ; Yunzhao WU ; Yun SUN ; Yingli WU
Journal of Shanghai Jiaotong University(Medical Science) 2017;37(4):432-437
Objective·To establish a cell-based screening system for identification of compounds with activity in regulating retinoic acid receptor (RARα) stability. Methods·The modified pMSCV plasmid constructs, named as RARα-EGFP-IRES-DsRed, consists of enhanced green fluorescent protein (EGFP) fusing to RARα and red fluorescent protein (DsRed) as internal references incorporating the internal ribosome entry site (IRES) as interval sequence. The RARα-EGFP-IRES-DsRed plasmid was stably transfected into NB4 cells which were named as NB4-pMGIR-RARα. Fluorescence signals of EGFP and DsRed indirectly reflecting the expression of RARα, were detected by flow cytometry in cells that were treated with all-trans retinoic acid, sodium valproate, cytarabine, lenalidomide, etoposide, montelukast and gambogic acid, respectively. Effects of these compounds on the expression of RARα protein were further examined by Western blotting. Results·A double fluorescence reporter system for screening compounds that can increase the stability of RARα protein was successfully established, and sodium valproate was identified as a potent compound to promote the stability of RARα. Conclusion·The double fluorescence reporter system can be used to screen compounds regulating the stability of RARα protein, which can be further used to identify compounds regulating the stability of other proteins.
7.Stem cells protein microarray analysis after chicken egg-white extract induced cells
Guangping RUAN ; Xiang YAO ; Jufen LIU ; Jinxiang WANG ; Zian LI ; Yuanyuan HU ; Xinghua PAN
Chinese Journal of Immunology 2015;(9):1217-1220
Objective:To analyze the chicken egg-white extracts were co-cultured with cells whether elevated stem cells protein,whether the cells transformation into stem cell.Methods:Four kinds of cells,making a common culture,a 50% chicken egg-white extract co-cultured for 3 days,cells were collected and frozen at -80 degrees,sending the company to do stem cell protein microarray.Results:C57-BMSC has three proteins occurred statistically significant change , TS-UC-MSC has one proteins occurred a statistically significant change ,293T has one protein occurred a statistically significant change ,and 293T-GFP has one protein occurred a statistically significant change.Conclusion:50% chicken egg-white extract co-cultured cells,the cells occurred the phenomenon of transformation into stem cells.
8.Change of motor neurons and skeletal muscles distal to the lesion after spinal cord injury in rats
Yuanyuan WANG ; Yi HONG ; Xuefei WANG ; Qing CAI ; Hehu TANG ; Xiang LI ; Guangxu LIANG ; Junwei ZHANG
Chinese Journal of Tissue Engineering Research 2014;(33):5323-5328
BACKGROUND:The majority of studies focus on the lesions of spinal cord injury, while little evidence is available on the change of morphology and structure of distal nerve, muscle and motor endplates fol owing spinal cord injury.
OBJECTIVE:To investigate the time window change of the morphology of motor neurons and skeletal muscles caudal to the lesion after spinal cord injury in rats.
METHODS:Fifty healthy adult Sprague-Dawley rats were randomly divided into three groups:control group (n=5;without treatment), sham operation group (n=10), and spinal cord injury group (n=35). The sham operated rats only received laminectomy. In the spinal cord injury group, rats were subject to complete T 10 spinal cord injury by total laminectomy and cord transverse resection. Then the morphological change including sciatic nerve, motor endplate and median gastrocnemius was observed for each group at 1, 2, 4, 12, 24 weeks after injury.
RESULTS AND CONCLUSION:(1) The myelin sheath layers of sciatic nerve were separated partial y at 4 weeks in rats with spinal cord injury, the myelin sheaths were fragmented with the regeneration of thin-myelinated and unmyelinated axons at 12 weeks. There was a decrease in myelinated axons and an increase in thin-myelinated and unmyelinated axons at 24 weeks. (2) The synaptic gutters of motor endplate, the presynaptic and postsynaptic membrane and synaptic space were distinct at 4 weeks in rats with spinal cord injury, the degenerated motor endplates coexsisted with the intact ones at 12 weeks. The motor endplate disappeared at 24 weeks. (3) There was a slight decrease in muscle cross-sectional area at 2 weeks in rats with spinal cord injury, but no structural change was found, the membrane of myocytes was partial y weakened at 4 weeks, the border of myocytes was obscure with hyperplasia of connective tissue at 12 weeks, and myocytes gathered and in fusion at 24 weeks. As natural history of completely transected spinal cord injury in rats, there were significant changes in morphology of peripheral nerve, motor endplate and skeletal muscles caudal to the lesion at 12 weeks, and the changes were destructive at 24 weeks.
9.Study of mechanism of DXM and NAC inhibiting expression of IL-8 and ICAM-1 in A549 cells
Qi XIANG ; Xin FU ; Pixin RAN ; Jin ZHANG ; Xiwei ZHENG ; Juan CHEN ; Yuanyuan GUO
Chinese Pharmacological Bulletin 2014;(9):1293-1297,1298
Aim To study the mechanism of DXM and NAC inhibiting the expression of IL-8 and ICAM-1 in A549 cells. Methods The expression of IL-8 and ICAM-1 was detected by ELISA and flow cytometry re-spectively; the expression of GR,HDAC,AP-1,NF-κB was detected by Western blot, while the activity of HDAC was detected by spectrophotometry. ResultsThe increasing expression of IL-8 and ICAM-1 induced by TNF-α could be inhibited by DXM and NAC in A549 cells. DXM could inhibit the transcribed activa-tion of AP-1,NF-κB, and the expression of HDAC and its activity induced by TNF-α and LPS; NAC only in-hibited the transcribed activation of NF-κB, while it had no affection on the transcribed activation of AP-1 and the expression of HDAC and its activity. Conclu-sions DXM and NAC both have the anti-inflammatory effect. DXM plays the role of anti-inflammation through increasing the expression and activation of HDAC, in-hibiting the transcribed activation of AP-1 and NF-κB, while NAC has no effect on the expression and activa-tion of HDAC, which shows that NAC does not exert anti-inflammatory effect through acetylation signal.
10.The bithermal test in idiopathic benign paroxysmal positional vertigo.
Xin MA ; Xiang ZHANG ; Yuanyuan JING ; Lisheng YU
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2009;23(10):439-442
OBJECTIVE:
Comparison of the bithermal test in benign paroxysmal positional vertigo (BPPV) before and after particle repositioning maneuver (PRM), identify the possible mechanisms of canal paralysis in BPPV.
METHOD:
Ninety-six BPPV cases during 2005-2008 were randomly divided into two groups: 52 cases were carried bithermal test before PRM, and the other 44 cases were carried after successful PRM. According to the canal paresis(CP) > or = 20%, subdivided each patients group the normal semicircular canal (SC) function group and the paresis SC function group. Compared the ratio of paresis SC function between the subgroups.
RESULT:
Except the ages of the first group were younger than the second group, the gender rate, semicircular canal involved, side involved, course of the disease, recurrent attacks were all equal between the two groups. Using chi square test to compare the ratio of paresis SC between the two groups (t = 0.654, P < 0.05), the differences between the two groups were not statistical.
CONCLUSION
PRM has no influence to the bithermal test in BPPV. The mechanisms of the paresis SC function may be the extensive pathological changes in the vestibular system. The floating otolith has little effect on the dynamic of the endolymph.
Adult
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Aged
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Aged, 80 and over
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Caloric Tests
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Female
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Humans
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Male
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Middle Aged
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Otolithic Membrane
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Paresis
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physiopathology
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Semicircular Canals
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physiopathology
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Vertigo
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physiopathology
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Young Adult