2.Construction and expression of Runx1-shRNA expressiong plasmid.
Yifan CHEN ; Xiaoju TANG ; Fengming LUO
Journal of Biomedical Engineering 2014;31(4):837-841
This study aims to construct the plasmid of human Runx1 and observe its possible effects on Runx1 gene expression in human pulmonary adenocarcinoma cells (A549). The shRNA sequence targeting human Runx1 was designed and synthesized, then inserted into pSuper plasmid by DNA recombination technology. The recombinant plasmid was confirmed by bacterial colonies PCR, enzyme digestion analysis and DNA sequencing. A549 cells were transfected by Runx1 shRNA plasmid. The inhibition efficiency of pSuper-Runx1-shRNA plasmid on Runx1 at mRNA level and protein level were measured with real-time PCR and Western blot. The results of real-time PCR and Western blot indicated that the mRNA and protein levels of Runx1 in A549 cells were inhibited by the pSuper-Runx1- shRNA expression plasmid, and the inhibition rate were 33% and 50%, respectively.
Cell Line, Tumor
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Core Binding Factor Alpha 2 Subunit
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genetics
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Gene Expression
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Genetic Vectors
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Humans
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Plasmids
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genetics
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RNA, Messenger
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genetics
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RNA, Small Interfering
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genetics
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Real-Time Polymerase Chain Reaction
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Transfection
4.Prevention of skin flap necrosis following eradicative operation of breast cancer
Guolin CHENG ; Fengjun WANG ; Yingwei XUE ; Yifan ZHANG ; Xiangqan LUO
Chinese Journal of Practical Surgery 2001;21(4):228-229
Objective To study how to reduce the incidence rate of skin flap necrosis after the eradicative resection of breast cancer. Methods 45 patients with breast cancer underwent eradicative operation with new methods to prevent the necrosis of skin flap,and they were studied and compared with 1210 cases treated by traditional ways. Results With the new methods adopted, the necrosis rate of skin flap following eradicative operation of breast cancer was reduced from 45%(original)to 2%(present).Statistic analysis showed that there was outstanding difference between them. Conclusion In the eradicative operation of breast cancer,the key to prevent the necrosis rate of skin flap is:prevention of subaxillary lymphatic fistula, two tubular drainage set individually in subcostalis and subaxillaris, appropriate force of chest bandaging, proper thickness of skin flap and perfect suture without tension.
5.Clinical Dosage Regimens of Voriconazole Evaluated by Monte Carlo Simulation
Yifan LUO ; Lixiang REN ; Qi SUN ; Mingyan JIANG
China Pharmacy 2017;28(14):1907-1911
OBJECTIVE:To provide reference for clinical individual medication of voriconatole. METHODS:The distribution of MIC of voriconazole to Aspergillus fumigatus and Candida albicans were summarized as well as the pharmacokinetic parameters of voriconazole in different populations. Using probability of target attainment(PTA)and cumulative fraction of response(CFR)as indexes,crystal ball software 11.1.2.4 was used for Monte Carlo simulation of different dosage regimens of same population and same dosage regimen of different populations. RESULTS:For children with impaired immunity,when the drug doses of were 4,6 mg/kg and MIC was lower than 0.125 mg/L,PTA was higher than 90%;when the drug doses was increased to 8 mg/kg and MIC was lower than 0.125 mg/L,PTA was higher than 90%. For different populations receiving same dosage regimens(4 mg/kg),MIC of teenagers with impaired immunity was lower than 0.25 mg/L and those of healthy adults,patients underwent hematopoietic stem cell transplantation and adults with impaired immunity were all lower than 0.5 mg/L,PTA was higher than 90%. CFR to A. fumigatus were 42.53%,58.41%,77.74%,70.16%,89.40%,93.72%,95.42% and CFR to C. albicans were 96.68%,97.13%,97.94%, 97.54%,98.07%,98.28%,98.35%among children with impaired immunity receiving different drug doses(4,6,8 mg/kg)and dif-ferent populations receiving drug dose of 4 mg/kg(teenagers with impaired immunity,healthy adults,patients underwent hemato-poietic stem cell transplantation,adults with impaired immunity). CONCLUSIONS:Various dosage regimens of different popula-tions included in this study could effectively control C. albicans infection. It is necessary to increase the drug dose of children and teenagers with impaired immunity in order to meet the needs of A. fumigatus infection treatment.
6.Experimental Study of Antidepressant Effects of Total Timosaponin
Lixiang REN ; Yifan LUO ; Shaojiang SONG ; Xia LI ; Yingliang WU
Traditional Chinese Drug Research & Clinical Pharmacology 1993;0(01):-
Objective To study the antidepressant effect of total timosaponin(TT)and its mechanism.Methods The antidepressant effect of TT was examined by mice forced swimming test(FST),learned helplessness(LH)experiment,chronic mild stress(CMS)model,yohimbine induced lethality test and 5-HTP induced head-twitches test.Results TT(25 or 50 mg?kg-1,ig,qd?14 d)markedly shortened the immobility time in the FST,but didn't affect the autonomic activity.TT(12.5,25 or 50 mg?kg-1,ig,qd?14 d)significantly decreased the number of escape deficits in the LH mice.TT(25 or 50 mg?kg-1,ig,qd?21 d)markedly enhanced the locomotor activity and increased consumption of sucrose solution in CMS mice.TT(50 mg?kg-1,ig,qd?14 d)enhanced the mortality of mice after administration of yohimbine for 4 h,and distinctly increased the head-twitch number in the 5-HTP induced head-twitches test.Conclusion TT has antidepressant effects in various depression mouse models.Its mechanism may be related to the reinforcement of NE and 5-HT nerves system.
7.Meta-analysis of effectiveness of Chinese materia medica as adjuvant therapy of antibacterial for pediatric mesenteric lymphadenitis
Yifan LUO ; Lixiang REN ; Xiaohong LI ; Mingyan JIANG
Drug Evaluation Research 2017;40(6):851-858
Objective To assess the effectiveness of Chinese materia medica (CMM) as adjuvant therapy of antibacterial for pediatric mesenteric lymphadenitis.Methods Retrieved RCT research of CMM as adjuvant therapy of antibacterial for pediatric mesenteric lymphadenitis from CNKI,Wanfang,VIP,PubMed,and Medline database from 1980 to 2016.Literatures were selected according to the inclusion and exclusion crteria,Jadad scores method was used to evaluate the literatures and meta-analysis was performed by RevMan 5.3.Results A total of 34 randomized controlled trials were included.Meta-analysis showed that the trial group was superior to the control group in terms of efficiency,cure rate,remission time of main symptoms,mesenteric lymph node recovery,and recurrence rate,there was statistical difference.Conclusions CMM as adjuvant therapy of antibacterial for pediatric mesenteric lymphadenitis is more effective than the use of antibacterial alone.
8.Summary of research methods for drug intestinal absorption
Yifan LUO ; Lixiang REN ; Xiaohong LI ; Yaoting SUN ; Mingyan JIANG
Drug Evaluation Research 2017;40(5):706-710
At present,the study of intestinal absorption of oral drugs mainly includes in vitro,in vivo and in situ methods.In view of the advantages of in situ intestinal perfusion such as simple operation,mature technology,controllable,ensure the neuroendocrine regulation and blood supply,and so on,which could better reflect the true situation of drug absorption.In this study,the research methods and characteristics of intestinal absorption of oral drugs were systematically introduced.The recirculating perfusion method and single-pass perfusion method were compared,and several volume correction methods were also introduced.In order to ensure the operability and accuracy of experimental results,proper experiment method of intestinal absorption will be adopt according to the factors such as drug characters,experiment requirements,experimental conditions,and so on.The article provides a scientific basis for the development of pharmaceutical dosage and clinical rational drug use.
10.Application of OTSU-based self-attenuation correction PET reconstruction technology in 18F-AV45 imaging
Xinchong SHI ; Lifei YAN ; Yifan ZHENG ; Chang YI ; Ganhua LUO ; Bo ZHANG ; Xiangsong ZHANG
Chinese Journal of Nuclear Medicine and Molecular Imaging 2021;41(1):18-22
Objective:To explore the application of OTSU-based self-attenuation correction PET (sacPET) reconstruction technology in 18F-florbetapir (AV45) imaging. Methods:From November 2018 to December 2019, 7 confirmed Alzheimer′s disease (AD) patients (4 males, 3 females, age (69.6±4.5)years) and 3 healthy controls (HC; 1 male, 2 females, age (68.0±4.6) years) were recruited prospectively for 18F-AV45 PET imaging in the First Affiliated Hospital of Sun Yat-Sen University. Original data collected by PET acquisition was processed with sacPET reconstruction and then compared with standard PET images by visual analysis and semi-quantitative analysis. Fisher exact test, Kappa test and Pearson correlation analysis were used to analyze data. Results:In HC group and AD group, the radioactive distribution showed by sacPET images and that by standard PET images were similar, and the contrast of gray-white matter in sacPET images was weaker than that in standard PET images. Moreover, the positive uptake area of the cortex in the AD group was smaller than that in standard PET images. Visual analysis showed 19 positive regions in sacPET images and 22 in standard PET images, with no statistical difference of positive rates of the sub-regions in the cortex between the two PET images (all P>0.05), and the overall consistency of 88.00% (44/50; Kappa=0.75 (95% CI: 0.57-0.94), P<0.05). Semi-quantitative analysis showed that the standardized uptake value ratio (SUVR) of frontal lobe and cingulate gyrus measured by sacPET was lower than that measured by standard PET (0.93±0.06 vs 0.96±0.06 and 0.99±0.04 vs 1.01±0.04; t values: 5.30 and 5.10, both P<0.01), while SUVR of parietal lobe, temporal lobe and occipital lobe measured by sacPET was higher than that measured by standard PET (0.78±0.08 vs 0.68±0.07, 0.97±0.07 vs 0.91±0.08 and 0.94±0.11 vs 0.71±0.12; t values: 6.27, 7.36 and 16.90, all P<0.01). The overall SUVR of sacPET images was significantly correlated with the standard PET images ( r=0.75, P<0.001). Conclusion:For 18F-AV45 imaging, sacPET reconstruction technology can obtain reliable and effective PET images without CT data, but its accuracy and precision still need to be improved.