1.Ultrasound mediated microbubble destruction enhances antisense c-myc expression in liver cancer cells SMMC-7721 in vitro
Hui JING ; Wenjia GUO ; Wen CHENG
Chinese Journal of Ultrasonography 2009;18(9):809-812
Objective To develop a novel approach of transfecting the exogenous gene safely and effectively mediated by receptor according to the cavitation effect of ultrasound-mediated microbubble destruction. Methods C-myc antisense oligonucleotide(ASODN) and galactosylation polylysine(G-PLL) which was fluoresce in-labeled specificity ligand and SonoVue microbbule were coupled by electrostatic interaction. SMMC-7721 cell with c-myc ASODN were transfected in vitro by ultrasound-mediated microbubble destruction to investigate the effect of c-myc ASODN on it. The c-myc mRNA gene expression of liver cancer SMMC-7721 cells was detected. Results Cultured with 10% SonoVue microbubble and under ultrasound at 1.5 MHz,1.0 W/cm2, 10% duty cycle,60 s respectively,the SMMC-7721 cells had not been changed significantly. Reverse transcription polymerase chain reaction(RT-PCR) assay showed that c-myc expression was decreased in transfected cells(P<0.01). Conclusions Receptor-mediated targeted microbubble destruction by ultrasound can enhance the exogenous gene transfection and expression. It is a promising method in gene therapy of liver cancer.
2.Effect of high glucose on cultured retinal Müller cell in vitro
Jing, GUO ; Min, KE ; Xiao-feng, WEN
Chinese Journal of Experimental Ophthalmology 2012;30(9):791-794
Background Retinal Müller cells can offer nutrient and maintain the normal structure of retina.Researches showed that the abnormality of Müiller cells leads to retinal vascular disease.To explore the effect of high glaucoma on retinal Müller cells is of a very important significance for the study on diabetic retinopathy (DR).Objective This study was to investigate the effects of different concentrations of glucose on retinal Müller cells in vitro.Methods Retinal tissue was isolated from 1 10-day-oM clean SD rat.Mtiller cells were cultured by explant culture method and passaged in DMEM containing 20% fetal bovine serum.The third generation of cells were obtained and identified using glial fibrillary acidic protein (GFAP) staning.Then,5.5,30.0 and 40.0 mmol/L glucose were added into the culture medium for 4 days respectively.The proliferation (A570) of Müller cells was detected by MTT,and apoptosis rate of Müller cells was calculated by flow cytometer to evaluate the effects of 5.5,30.0 and 40.0 mmol/L glucose to cell vitality.Results Cultured and passaged cells grew well with the spindle shape.The positive reactive cells were >95% for GFAP.The A570 value of Müller cells was 0.24±0.01,0.21±0.03 and 0.20±0.02 in 5.5,30.0 and 40.0 mmol/L glucose group respectively,showing a significant difference among the three groups(F=6.755,P<0.05).Compared with 5.5 mmol/L glucose group,As70 values were significantly lower in 30.0and 40.0 mmol/L glucose group (q =0.645,0.486,P < 0.05).Apoptosis rates of Miiller cells were (26.40 ±0.25)%,(30.19±0.16)% and (36.23±0.19)% in 5.5,30.0 and 40.0mmol/L glucose groups,with a significant difference among them (F =294.530,P<0.05),and those in 30.0 and 40.0 mmol/L glucose groups were significantly reduced in comparison with 40.0 mmol/L glucose group (q =0.754,0.484,P < 0.05).Conclusions High concentration of glucose inhibits the viability and promote the apoptosis of retinal Müller cells at a concentrationdependent manner.
3.The Normal Anatomy and CT Study of HDL
Yajuan WEN ; Jing MA ; Feng WANG ; Guangfu YANG ; Youmin GUO
Journal of Practical Radiology 2000;0(02):-
Objective To study the normal anatomy and pathologic CT findings of HDL and compare the HDL dimension of two conditions.Methods 160 consecutive CT scans of the normal upper abdomen were reviewed and 19 cases of cadavers were used.Results There was no statistics differences between the cadavers and normal subjects on HDL.Conclusion The real dimension of HDL can be replaced by the measurement on CT image,CT is a accurate means of measuring HDL in vival of human being.
4.Screening and identification of metastasis-related gene in salivary gland adenoid cystic carcinoma with cDNA microarry assay
Xiuli ZHU ; Junzheng WU ; Desheng WEN ; Qingyu GUO ; Jing WANG
Journal of Practical Stomatology 1995;0(04):-
Objective:To identify metastasis-associated genes in salivary gland adenoid cystic carcinoma(ACC).Methods:Salivary gland adenoid cytic carcinoma cell line ACC-2 and its highly metastatic ACC-M cells were used to screen the metastasis-related genes in ACC by microarray technology.Two fluorescent cDNA probes labeled with Cy3 and Cy5 dyes,were prepared from the mRNA samples of ACC-2 and ACC-M cells by reverse transcription method.The two color probes were then mixed and hybridized on the cDNA chip constructed by double dots of 1152 human genes,and scanned at two wave lengths.Differentialy expressed genes of the two cell lines were analyzed using computer.Then seven of the differently expressed genes were further validated by RT-PCR technique.Results:Of the 1,152 known genes and expressed sequence tags,26 showed significantly different expression level(minimum 2 fold) between the two cell lines.Among the 26 genes,19 were up regulated(with ratio more than 2) and 7 were down(with ratio less than 1/2).The results of RT-PCR analysis for 7 differently expressed genes were coincident with those of microarray assay.Conclusions:Down regulation of LIFR,LCP1,DPEP1 and ABLIM1,and up regulation of DCC,MMP1 and CNTN2 may be related to the highly metastatic potential of ACC-M cell line.
5.Treating diabetes mellitus from liver
Shanshan JING ; Zhimin AI ; Wen SUN ; Tonghua LIU ; Xiangyu GUO
International Journal of Traditional Chinese Medicine 2012;(12):1095-1097
Depending on the meaning and function of the liver in traditional Chinese medicine and modern medicine,this paper explores the important role of liver in the pathogenesis of DM (Diabetes Mellitus,DM).Combined with treatment based on syndrome differentiation and specific case report study,this paper has stressed the importance of treating DM from liver and so to set a sound basis for the systemic treatment based on syndrome differentiation from internal organs for DM.
6.Emphasizing clinical investigation on anti-inflammatory therapy of dry eye
Zu-guo, LIU ; Jing, LIU ; Wen-zhao, YANG
Chinese Journal of Experimental Ophthalmology 2013;(5):417-420
Inflammation is a defensive reaction and the most common pathological manifestation of dry eye.In addition,excessive inflammatory response is considered to be the most common pathogenic factor and main cause of dry eye.Currently,the active mechanism of anti-inflammatory drugs has been well-known,and topical antiinflammatory therapy for dry eye is exerting a role at certain extend.However,some adverse responses of these drugs are emerging during the treating procedure.Therefore,it is emphasized that a large sample size of and multicenter randomized-controlled clinical trial is needed to identify the different effects of various anti-inflammatory drugs for different types of dry eye diseases,which will offer a basis for standardized anti-inflammatory treatment for dry eye.
8.Design, synthesis and pharmacological investigation of isoindoline derivatives as 5-HT/NE double reuptake inhibitors.
Hui WEN ; Yuan SHI ; Jing-wen DONG ; Yan-shen GUO ; Jian-Jun ZANG ; Guang-zhong YANG
Acta Pharmaceutica Sinica 2015;50(9):1148-1155
A series of isoindoline derivatives were designed, synthesized, and evaluated for their double inhibitory activities. All of them were new compounds, and their structures were confirmed by 1H NMR and HR-MS. Preliminary in vitro pharmacological tests showed that all compounds exhibited 5-HT or NE reuptake inhibition activity. Among the tested compounds, compound I-3 exhibited potent inhibitory activity against 5-HT and NE reuptake in vitro, and exhibited potent antidepressant activity in vivo. These compounds designed can be further optimized for finding more potent 5-HT/NE dual reuptake inhibitors and antidepressant candidates as well.
Antidepressive Agents
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chemical synthesis
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chemistry
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Biological Transport
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Drug Design
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Isoindoles
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chemical synthesis
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chemistry
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Serotonin Uptake Inhibitors
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chemical synthesis
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chemistry
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Structure-Activity Relationship
9.Expressions and significance of miRNA - 101,enhancer of ZESTE homolog 2 and transforming growth factor-β1 in renal obstruction models in young rats
Jing CHANG ; Zhan GUO ; Lingang CUI ; Chuanchuan REN ; Xinmei GAO ; Shuman HUANG ; Bing BAI ; Jianguo WEN
Chinese Journal of Applied Clinical Pediatrics 2015;(21):1656-1659
Objective To explore the expressions and significance of miRNA - 101,enhancer of ZESTE homo-log 2(EZH2)and transforming growth factor(TGF)- β1 in the kidneys with complete uniateral ureteral obstruction (CUUO). Methods Thirty male SD rats,(6 ± 1)weeks old,weighted(150 ± 10)g,were divided into sham group, 7 - day group with CUUO and 14 - day group with CUUO by using random number table method,10 rats in each group. The obstructed kidney samples were collected in 7 and 14 days,respectively,for detecting the expression of miRNA -101 by real time - polymerase chain reaction(RT - PCR)and TGF - β1 and EZH2 protein by Western blot,immuno-histochemistry and hematoxylin - eosin staining. Their correlated expressions were analyzed. Results RT - PCR results showed that the expressions of miRNA - 101 in sham group and 7 - day group with CUUO were(12. 69 ± 1. 60)times and(3. 74 ± 1. 24)times which were higher than those of 14 - day group with CUUO,respectively,there was a signifi-cant difference among these 3 groups(P ﹤ 0. 05). The expressions of TGF - β1 and EZH2 proteins were 1. 14 ± 0. 12, 1. 04 ± 0. 14,0. 76 ± 0. 18 and 1. 04 ± 0. 04,0. 89 ± 0. 03,0. 73 ± 0. 02 in 14 - day group with CUUO,7 - day group with CUUO and sham group,respectively. There was a negative correlation between miRNA - 101 with EZH2( r =- 0. 92,P ﹤ 0. 05),and negative correlation with TGF - β1(r = - 0. 63,P ﹤ 0. 05),and positive correlation between EZH2 and TGF - β1(r = 0. 67,P ﹤ 0. 05);the expressions of miRNA - 101,EZH2 and TGF - β1 were associated with each other in obstruction renal in different time periods. Conclusions With the extension of obstruction time, miRNA - 101 expression decreased,EZH2 and TGF - β1 expression increased,evidently,which indicates that the de-velopment of renal interstitial fibrosis may be affected through regulating EZH2 expression of renal obstruction through miRNA - 101 in the young rats.
10.Determination of Au and Ag in Geological Samples by Loaded Polyurethane Foam-Inductively Coupled Plasma-Mass Spectrometry
Xianglei LIU ; Wenjun SUN ; Tianyao WEN ; Tengfei WANG ; Weizhi SUN ; Yongxin LI ; Jing GUO
Chinese Journal of Analytical Chemistry 2015;(9):1371-1376
The relatively high abundance of geochemical elements such as Nb, Zr, Y and other elements shows serious interferences in the determination of trace silver in geochemical samples by inductively coupled plasma-mass spectrometry ( ICP-MS) . Thus it will lead to large deviation in the determination of geochemical samples without separation and enrichment. The traditional emission spectrum or graphite furnace atomic absorption method is only single-element analysis to the silver and with bad sample representativeness. In this study, load diphenylthiourea ( DPTU) foam selective enrichment was used for the separation of Au and Ag from other interfering elements in geological samples, and thiourea liberation-ICP-MS method was developed for the simultaneous determination of Au and Ag. The samples were first decomposed by 1:1 aqua regia. After addition of 50 mL of water, the samples were adsorbed under oscillation for 30 min at 20℃. The detection limits of the Au and Ag were 0. 02 ng/g and 0. 007μg/g, respectively. The proposed method was successfully applied to the determination of Au and Ag in eight national standard materials.