1.The quantitative assessment of left ventricular regional diastolic function after intracoronary stent implantation by strain rate imaging
Yanfeng DUAN ; Lingling WEI ; Zhen TAO ; Yi HUANG
Chinese Journal of Postgraduates of Medicine 2014;37(7):40-43
Objective To investigate the application of strain rate imaging in quantitative assessment of left ventricular regional diastolic function after intracoronary stent implantation.Methods Fifty-six healthy person and 60 patients with coronary artery disease were performed quantitative assessment of left ventricular regional diastolic function by apical four chamber view,apical two chamber view and left ventricular long axis view before and after intracoronary stent implantation using strain rate imaging.Results Compared with those of normal myocardium,the peaks of strain rate curve at diastole of ischemic myocardium were reduced,and were increased after operation (P < 0.05).Conclusions Intracoronary stent implantation can significantly improve the blood supply to the ischemia myocardium.Strain rate imaging can quantitatively analyze the changes of left ventricular regional diastolic function.
2.Preparation and purification of siRNA targeting a proliferation-inducing ligand of pancreatic cancer cell line
Zhen-Biao MAO ; Wei-Yi WANG ; Jie-Fei HUANG ;
Academic Journal of Second Military Medical University 2000;0(08):-
Objective:To prepare and purify siRNA targeting a proliferation-inducing ligand targeted(APRIL-siRNA),so as to provxde a basis for studying the role of APRIL in human pancreatic cancer.Methods:pET-22b-APRIL was constructed to express APRIL dsRNA of human pancreatic cancer cell line CFPAC-1 in E.coli and the product was purified by chromatography using CF-11 column.APRIL dsRNA was digested by RNaseⅢto prepare APRIL siRNA,then the reaction mixture was loaded onto a DEAE ion exchange chromatography to remove RNaseⅢfrom oligonucleotides,and size exclusion chromatography was used to purify 21 bp siRNA.The purified APRIL siRNA was used to transfect Chinese hamster ovary(CHO)cells and the expression of APRIL in CHO cells was observed under fluorescence microscope Results:APRIL dsRNA was successfully expressed in E.coli after IPTG induction and was purified by CF-11 column.dsRNA was hydrolyzed with RNaseⅢand was purified by DEAE ion exchange chromatography and size exclusion chromatography.15% nondenaturing PAGE and 12% SDS- PAGE confirmed that RNaseⅢwas removed from oligonucleotides and 21 bp siRNA was purified with size exclusion chromatography.It was also found that APRIL siRNA obviously depressed APRIL expression in CHO cells.Conclusion:We have successfully constructed APRIL siRNA targeting APRIL gene of CFPAC-1 cells with in vitro transcription,which provides a basis for knock-down of APRIL gene in CFPAC-1 cells.
3.Cloning and High Expression Anabaena Class-II Fructose-1, 6-bisphosphate Aldolase Gene in Escherichia coli
Lan-Zhen WEI ; Yi-Wen CUI ; Wei-Min MA ; Quan-Xi WANG ;
China Biotechnology 2006;0(05):-
The complete genomes of more cyanobacterial strains have been completed for sequence, and genetic engineering of cyanobacteria has evolved in the post-genome era. Since Kaneko and colleagues had completed the sequence for the complete genome of Anabaena sp. PCC 7120 in 2001, functions of some genes in this genome, including fructose-1,6-bisphosphate aldolase (FBA) gene, have been predicated using the method of bioinformatics. However, little information is available regarding whether this gene can encode FBA and its product characteristics of related enzyme. Here, to explore this information, the predicted II-FBA gene-encoding region in Cyanobase database was cloned by PCR method and then ligated into pET-32a to generate the expression vector, pET-FBA-II. The results of SDS-PAGE indicated that the expression level of the expected target polypeptide was approximate 23.4 percent as compared to total protein and the molecular weight is about 40 kDa as compared to the protein molecular marker. The results of enzyme activity analysis showed that the activity of II-FBA was ~11.8 U per mg protein and owned a standard activity of II-FBA. To sum up, the results not only prove the functional prediction of this II-FBA gene from the Cyanobase database, but also provide the important conditions for further studying its physiological and biochemical characteristics and functions of the gene expression product.
5.Research on immune-modulation effects and its mechanisms of Guizhi Fuling capsule and its active ingredient combination.
Yi-ping JIANG ; Xiu WU ; Na LI ; Liang CAO ; Gang DING ; Zhen-zhong WANG ; Wei XIAO
China Journal of Chinese Materia Medica 2015;40(15):3068-3071
The effects of Guizhi Fuling capsule and its active ingredient combination within different concentration on SPL proliferate were observed by MTT method. The ratio of CD80/86, CD3CD25 and CD3CD69 was used to evaluate cell activation effects of Guizhi Fuling capsule and its active ingredient combination by FCM. Guizhi Fuling capsule with concentration of 400 mg · L(-1)can promote spleen lymphocyte proliferation, as well as the active ingredient combination, which showed the obvious dose-effect relationship. Compared with control group, the difference has statistical significance (P≤0.01). The result of FCM showed that Guizhi Fuling capsule and its active ingredient combination can promote CD80 and CD86 expression on spleen lymphocyte, and also can increase CD25 and CD69 ratio between spleen CD3+ cells. Compared with control group, the difference has statistical significance (P≤0.01). Thus, Guizhi Fuling capsule and its active ingredient combination may have immune-modulate effects, and the mechanism may have a close relationship with the lymphocyte activation.
Animals
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Capsules
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Drugs, Chinese Herbal
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analysis
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pharmacology
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Immunologic Factors
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pharmacology
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Lymphocyte Activation
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drug effects
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Male
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Mice
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Mice, Inbred BALB C
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T-Lymphocytes
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drug effects
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immunology
6.Optimization of extraction technology for salidroside, tyrosol, crenulatin and gallic acid in Rhodiolae Crenulatae Radix et Rhizoma with orthogonal test.
Xin LUO ; Xue-jing WANG ; Yi-wu ZHAO ; Wen-zhe HUANG ; Zhen-zhong WANG ; Wei XIAO
China Journal of Chinese Materia Medica 2015;40(18):3590-3593
The extracting technology of salidroside, tyrosol, crenulatin and gallic acid from Rhodiolae Crenulatae Radix et Rhizoma was optimized. With extraction rate of salidroside, tyrosol, crenulatin and gallic acid as indexes, orthogonal test was used to evaluate effect of 4 factors on extracting technology, including concentration of solvent, the dosage of solvent, duration of extraction, and frequency of extraction. The results showed that, the best extracting technology was to extract in 70% alcohol with 8 times the weight of herbal medicine for 2 times, with 3 hours once. High extraction rate of salidroside, tyrosol, crenulatin and gallic acid were obtained with the present technology. The extracting technology was stable and feasible with high extraction rate of four compounds from Rhodiolae Crenulatae Radix et Rhizoma, it was suitable for industrial production.
Chemical Fractionation
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methods
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Chemistry, Pharmaceutical
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methods
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Coumarins
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isolation & purification
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Drugs, Chinese Herbal
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isolation & purification
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Gallic Acid
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isolation & purification
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Glucosides
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isolation & purification
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Phenols
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isolation & purification
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Phenylethyl Alcohol
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analogs & derivatives
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isolation & purification
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Rhizome
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chemistry
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Rhodiola
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chemistry
9.Stability of akebia saponin D in gastrointestinal contents of rats.
Can-Yi GONG ; Wei XIAO ; Zhen-Zhong WANG ; Zhon-Glin YANG
China Journal of Chinese Materia Medica 2014;39(12):2311-2313
In order to study the stability of akebia saponin D (ASD) in biological fluids in vitro, the determination methods of ASD were established in this study. Akebia saponin D was dissolved in artificial gastric juice, intestinal juice and gastrointestinal contents of rats, respectively, then thermostatically maintained at 37 degrees C. At time intervals after degradation, samples were withdrawn and the concentrations of ASD were determined by HPLC, from which stability of it at different biological specimen was evaluated. As a result, ASD was totally degraded in large intestinal contents of rats in 8 hours. ASD was very stable in artificial gastric juice, intestinal juice and gastric contents of rats. All of the above data proved that ASD was easily degraded by coliform bacteria but stable in acid environment and with the presence of digestive enzyme.
Animals
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Drug Stability
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Drugs, Chinese Herbal
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administration & dosage
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pharmacokinetics
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Gastrointestinal Tract
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chemistry
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metabolism
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Humans
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Rats
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Saponins
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administration & dosage
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chemistry
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pharmacokinetics
10.Effect of Acupuncture on Expressions of InsR-β mRNA and Protein in the Liver of Rats with Insulin Resistance
Jian SUN ; Nenggui XU ; Wei YI ; Zhen JIA ; Zhonghua YANG ; Xuejun CUI
Journal of Acupuncture and Tuina Science 2009;7(4):196-199
Objective: To observe the effect of acupuncture on the expressions of insulin receptor β (InsR-β) mRNA and protein in the liver of experimental rats with insulin resistance (IR). Method: Twenty-four rats were randomly divided into 3 groups, control group (n=8),model group (n=8) and acupuncture group (n=8). Rats in the control group were fed with conventional food, and the other rats were induced into insulin resistance model with high fat-sugar-salt food. Once model was induced successfully, rats in the control group were fed with conventional food continually, rats in the model group were fed with high fat-sugar-salt food continually, and rats in the acupuncture group were fed with high fat-sugar-salt food, and treated with acupuncture for 2 weeks. The expression of InsR-β mRNA was detected by real-time fluorescent quantitative RT-PCR, and the expression of InsR-β protein was detected by Western blot. Result: Expressions of InsR-β mRNA and protein in the model group were significantly lower than those in the control group (P<0.01), and those in the acupuncture group were higher than those in the model group (P<0.01, P<0.05). The expressions of InsR-β mRNA and protein between the acupuncture and control group had no significant difference. Conclusion:Acupuncture treatment can increase the expressions of InsR-β mRNA and protein in IR rats'liver to improve insulin resistance.