3.Modification,Expression and Purification of Human Endotoxin Binding Peptide Gene
Ya-Li SUN ; You-Sheng LIU ; Hai-Jie YANG ;
China Biotechnology 2006;0(03):-
Objective: To modify EBP(endotoxin binding peptide), clone and express the mutate of EBP gene and gain purified mEBP.Method: mEBPgene was cloned by PCR site-directed mutagenesis. PinpointXa-3/mEBP expression vector was designed to express human mEBP as a fusion protein in BL21 (DE3) pLysS. Digested engineering bacteria by lysozyme and collected inclusion bodies.Fusion protein was purified by Pinpoint TM Xa purification system and cleaved by factorXa,mEBP was purified by RP-HPLC. Results: Mutations at residues 5 and 18(Gln→Lys) was obtained by PCR site-directed mutagenesis, expressed and purified mEBP successfully.Conclusions: Obtaining of purified mEBP lay a foundation for its biological activity research.
4.Effect of Local Intramuscular Repeated Injection with Botulinum Toxin-A for Treatment of Spastic Cerebral Palsy
ya-jie, WANG ; bao-qin, GAO ; wei-li, YANG
Journal of Applied Clinical Pediatrics 2004;0(08):-
Objective To observe the curative effect of botulinum toxin A(BTX-A)repeated intramuscular injection for treatment of serious spastic cerebral palsy.Methods Thirty cases with serious spastic cerebral palsy received local repeated intramuscular injection. The interval time was 3 months. The dose of BTX-A was 4 U/kg.The muscle tone was assessed with the modified ashworth scale and range of motion with physical rating scale(PRS).The indexes were evaluated before injection and 3 months after the second injection. Paired-samples t-test was used in the statistic analysis.Result The muscle tone and PRS had remarkable improvement after second therapy with BTX-A(P
5.Study on Ultrasonic Follow-up on Atheriosclerosis in Familial Hypercholesterolemia
qiang, YONG ; zhi-an, LI ; jie, LIN ; lu-ya, WANG
Journal of Applied Clinical Pediatrics 2006;0(13):-
Objective To identify the arteriosclerosis (AS)changes in peripheral artery and abdominal aorta of patients with familial hypercholesterolemia(FH) during follow-up.Methods Seventeen patients of 6 FH [5 male and 12 female with average age of (16.12?6.65) years old],along with 17 subjects of matching sexes and ages with normal blood cholesterol as healthy control group,underwent examination by color doppler ultrasound,and changes of intima-media thickness (IMT) in peripheral artery and abdominal aorta,severity of stenosis,morphology,and function were observed.Results For 17 patients of FH,the total cholesterol(TC),low density lipoprotein cholesterol(LDL-C) in serum were higher significantly and high density lipoprotein cholesterol (HDL-C) was lower significantly than those in healthy control group(Pa0.05).The IMT of carotid aorta,subclavicular aorta,common abdominal artery,and common femoral artery in 13 patients were showing various degrees of increase,yielding an average of 2.9 mm.As patients aged,this phenomenon was reported to be more profound in common abdominal aorta and femoral artery.Conclusions Patients of FH show AS lesion in early stage and it worsened as they aged,from carotid arteries to common abdominal aorta and femoral artery.Color doppler ultrasound can be a non-invasive examination for monitoring the progress of AS in blood vessels in patients of diagnosed FH.
6.Pharmacokinetics of topically applied econazole nitrate nanoparticles in rabbit eye
Bo, WANG ; Li-ya, WANG ; Peng, CHEN ; Jun-jie, ZHANG
Chinese Journal of Experimental Ophthalmology 2012;30(8):677-680
Background Econazole nitrate is not effective as an antifungal eyedrop because of its poor intraocular permeability,therefore changing the formulation of econazole nitrate to improve its intraocular permeability become a critical point in the treatment of intraocular fungal infection. Objective The present study was to observe the penetration of 0.5% econazole nitrate nanoparticles in the corneas and aqueous humors following its topicaladministration. Methods Econazole nitrate nanoparticles were prepared by quasi-emulsion solvent diffusion.Characteristics and size of nanoparticles were examined with transmission electron microscope and laser scatteringmethod,respectively.Econazole nitrate nanoparticles drops (0.5% )was topically administered in 27 New Zealandwhite rabbits bilaterally,and aqueous humor and corneas were obtained after the application of the eye drops for 5,15,30,45,60,90,120,180,240 minutes respectively to detect the concentration of econazole nitrate with highperformance liquid chromatography (HPLC). The pharmacokinetic parameters were calculated with 3 p97pharmacokinetic computer software.The use of the animals followed the Regulation for the Administration of AffairsConcerning Experimental Animals by State Science and Technology Commission. Results The diameter of thenanoparticles was 50 nm with the round shape and encapsulation efficiency was 96.0%.Econazole nitrate nanoparticlesat the concentration of 0.5% could be rapidly separated with other elements by HPLC with a lowest quantitativeconcentration of 0.1 mg/L.The mean recovery rates of econazole nitrate nanoparticles were 98.09% in cornea and 99.66% in aqueous humor,respectively after topical administration.The peak levels of econazole nitrate nanoparticles in cornea and aqueous humor were achieved at 5 minutes after application ( cornea:40.620 μg/g± 7.756 μg/g;aqueous humor:0.504 mg/L±0.153 mg/L),and its half-life( t1/2 )in cornea and aqueous humor was 23.5 minutes and 18.6 minutes,respectively. Conclusions Econazole nitrate nanoparticles at 0.5% concentration can remain a feasible bioavailability in ocular tissue and therapeutic level in cornea and aqueous humor.
7.The impurity profiling of simvastatin and its tablets by UPLC-MS/MS.
Jie LI ; Hai-Wei HUANG ; Hong ZHANG ; Tao LI ; Ya-Qin SHI
Acta Pharmaceutica Sinica 2014;49(5):672-678
Investigation of simvastatin and its related substances was carried out using a reversed phase ultra performance liquid chromatography/tandem mass spectrometry method. The identification of impurities in simvastatin was performed with a triple-quadrupole mass spectrometer, with an electrospray ionization (ESI) source in the negative/positive ion mode. A total of 12 compounds were characterized in commercial samples, among which 2 impurities had never been reported. All the impurities were deduced based on the MS fragment pathways of simvastatin and the biosynthetic pathway of lovastatin. This work provides very useful information for quality control of simvastatin.
Chromatography, High Pressure Liquid
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Chromatography, Reverse-Phase
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Drug Contamination
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Hypolipidemic Agents
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chemistry
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Quality Control
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Simvastatin
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chemistry
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Spectrometry, Mass, Electrospray Ionization
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Tablets
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Tandem Mass Spectrometry
8.Effect of Yixintai Granule on mRNA and protein expression levels of AQP, in renal medulla of chronic heart failure rabbits.
Yun TANG ; Zhi-Hua GUO ; Ya LI ; Tao SUN ; Gang-Qiang WU ; Li LIU ; Jie DENG
Chinese Journal of Integrated Traditional and Western Medicine 2015;35(3):333-337
OBJECTIVETo explore the effect of Yixintai Granule (YG) on mRNA and protein expression levels of AQP2 in renal medulla of chronic heart failure (CHF) rabbits.
METHODSCHF rat model was established by ear marginal vein injection of adriamycin. Successfully modeled rabbits were divided into the model group, the high (8.4 g/kg), middle (4.2 g/kg), and low dose (2.1 g/kg) YG group, and the Furosemide group (2 mg/kg). Besides, a normal control group was set up. Equal volume of physiological saline was administered to rabbits of the model group and the normal control group by gastrogavage. YG at different doses was administered to rabbits of the 3 YG groups by gastrogavage. The intervention lasted for 4 weeks, once per day. After treatment the urine volume and pathomorphological changes of renal medulla tissue were observed. mRNA and its protein expression levels of AQP2 were detected.
RESULTSCompared with the normal control group, the urine volume decreased significantly, mRNA and protein expression levels of renal medulla AQP2 increased significantly in the model group (all P < 0.01). Compared with the model group, the urine volume increased significantly, and mRNA and protein expression levels of renal medulla AQP2 decreased significantly in all medicated groups (all P < 0.01). Compared with the low dose YG group, the urine volume significantly increased and the mRNA expression level of renal medulla AQP2 significantly decreased in the middle and high dose YG groups (all P < 0.01). The expression level of AQP2 protein significantly decreased in the high dose YG group (P < 0.01). Pathological changes of the renal medulla was the most obviously seen in the model group. But they were alleviated to various degrees in all medicated groups. They were more obviously attenuated in the middle and high dose YG groups.
CONCLUSIONYG could improve CHF possibly through down-regulating mRNA and protein expression levels of AQP2 in renal medulla, and elevating the urine volume.
Animals ; Aquaporin 2 ; genetics ; metabolism ; Chronic Disease ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; therapeutic use ; Heart Failure ; drug therapy ; metabolism ; RNA, Messenger ; metabolism ; Rabbits ; Rats, Sprague-Dawley
9.Advance on the Biotransformation of Bioactive Natural Leading Compounds
Ya-Jie TANG ; Yan LI ; Xiao-Ling XU ; Dong-Sheng LI ;
China Biotechnology 2006;0(09):-
Biotransformation of bioactive natural leading compounds is a kind of bioprocess in which the structure of the added bioactive natural leading compounds could be modified by biocatalysts(e.g.,enzyme,microbial,plant and animal cells) in order to produce high efficient and low toxicity compounds.The biotransformation purpose of the known bioactive natural leading compounds is to improve its efficiency,or reduce its toxicity,or improve its solubility and bioavailability.The trace and high-valued bioactive natural leading compounds also could be produced by the biotransformation,and the biotransformation of bioactive natural leading compounds is still helpful to study the mechanism of drug metabolism.The current focus of the biotransformation of bioactive natural leading compounds is on the compounds of steroid,quinine,flavone and terpene,and some important biotransformation process has been successfully screened out.Fundamental research should be done in the following fields,such as the biotransformation mechanism of bioactive natural leading compounds,biotransformation process engineering,and the efficiency evaluation of bioproducts produced by biotransformation.The latest biotechnology(e.g.,directed evolution of biocatalyst,combinatorial biotransformation,non-aqueous biotransformation,high throughput screening) should be introduced to the biotransformation of bioactive natural leading compounds,which will boost its fast development.
10.Determination of Berberine Hydrochloride in Shenshe Ointments by HPLC
Shan MIAO ; Yan LI ; Jie LI ; Linlin BI ; Ya ZHANG ; Xiaopeng SHI
China Pharmacist 2014;(9):1576-1577,1578
Objective:To establish the determination method for berberine hydrochloride in Shenshe ointments. Methods: The quantitative analysis of berberine hydrochloride in Shenshe osintments was carried out by HPLC with a Kromsal C18 chromatographic column (250 mm × 4. 6 mm, 5 μm), the mobile phase was acetonitrile-0. 1% phosphoric acid (49∶51) (adding 0. 2g sodium dode-cylsulphate into 100 ml solution) with the flow rate of 1. 0 ml·min-1 and the detection wavelength of 320nm, the temperature of col-umn was room temperature, and the injection volume was 20 μl. Results: The linear range was 0. 059 2-0. 296 0 g·L-1 with good correlation(r=0. 999 4). The average recovery was 99. 80% and RSD was 0. 24%(n=6). Conclusion: The established method is accurate, specific and reproducible, and suitable for the determination of berberine hydrochloride in Shenshe ointments.